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The aim of the study is to evaluate the treatment patterns, time to start treatment, and healthcare resources utilization (HCRU) of cervical cancer (CC) patients within the Brazilian public health system (SUS). This is an observational retrospective study using SUS administrative database (DATASUS). Data from January-2014 to December-2020 was gathered from patients with the ICD-10 C53 codes. From 2014 to 2020, 206,861 women were included, among whom 90,073 (43.5%) had stage information. Of staged patients, 60.7% (54,719) had advanced disease (stages III and IV) and the most performed treatments were chemoradiotherapy (CRT) (41.6%), surgery + CRT (19.1%), radiotherapy (RT) only (16.8%) and chemotherapy (CT) only (13.3%). The proportion of patients submitted to CT in advanced stages was higher than in non-advanced stages (I and II), in contrast to RT, which was more frequent in stage I than stage IV. Median time to initiate treatment surpassed two months in approximately 30% of the cases, regardless of stage. Conization was the most performed surgical procedure. The hospitalization rate per patient per month for stage IV was twice as high as stage I (0.05 [95%CI 0.05-0.05] and 0.11 [0.11-0.11], respectively). The same trend was observed for outpatient visits (0.54 [95%CI 0.53-0.55] and 0.96 [0.93-0.98], respectively). This study demonstrated a high proportion of advanced CC at diagnosis in Brazil. The treatment pattern showed that chemoradiotherapy was the most frequent regimen overall and the use of chemotherapy and HCRU increased with staging. These results could provide information to improve public policies towards access to prevention, diagnosis, and treatment of CC in Brazil., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: TMFC, MACSM, PMB, MACR, MN, LPLG and CPM are employees of Merck Sharp Dohme Farmacêutica Ltda, Sao Paulo, Brazil, who may own stock and/or hold stock options in Merck & Co., Inc., Rahway, NJ, USA. ANR is a scientific medical consultant who was paid by MSD Brazil. PCP and ACS were employees of IQVIA Brazil, which was contracted by MSD Brazil to conduct the study., (Copyright: © 2024 Campuzano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The genetically modified cotton DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 expressing Cry1Ac, Cry1F and Vip3Aa19 from Bacillus thuringiensis Berliner (Bt) has been cultivated in Brazil since the 2020/2021 season. Here, we assessed the performance of DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton expressing Cry1Ac, Cry1F and Vip3Aa19 against Helicoverpa armigera (Hübner), Helicoverpa zea (Boddie), and their hybrid progeny. We also carried out evaluations with DAS-21023-5 × DAS-24236-5 cotton containing Cry1Ac and Cry1F. In leaf-disk bioassays, DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 was effective in controlling neonates from laboratory colonies of H. armigera, H. zea and the hybrid progeny (71.9%-100% mortality). On floral bud bioassays using L2 larvae, H. zea presented complete mortality, whereas H. armigera and the hybrid progeny showed <55% mortality. On DAS-21023-5 × DAS-24236-5 cotton, the mortality of H. armigera on leaf-disk and floral buds ranged from 60% to 73%, whereas mortality of hybrids was <46%. This Bt cotton caused complete mortality of H. zea larvae from a laboratory colony in the early growth stages, but mortalities were <55% on advanced growth stages and on floral buds. In field studies conducted from 2014 to 2019, DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton was also effective at protecting plants against H. armigera. In contrast, a population of H. zea collected in western Bahia in 2021/2022 on Bt cotton expressing Cry1 and Vip3Aa proteins, showed 63% mortality after 30 d, with insects developing into fifth and sixth instars, on DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton. We conclude that H. armigera, H. zea, and their hybrid progeny can be managed with DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton; however we found the first evidence in Brazil of a significant reduction in the susceptibility to DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton of a population of H. zea collected from Bt cotton in Bahia in 2021/2022., Competing Interests: The authors have declared that no competing interests exist. Corteva Agriscience provided support in the form of salaries for authors Luiz H. Marques, Tamylin K. Ishizuka, Renata R. Pereira, Ademar N. Istchuk, Jaedino Rossetto, Valeria F. Moscardini, Oscar A. N. B. e Silva, Antonio C. Santos, Timothy Nowatzki, Mark L. Dahmer, Amit Sethi, Nicholas P. Storer, but did not have any additional role in the conduction of field trials, data collection and analysis. The specific role of these authors are articulated in the ’author contributions’ section. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Marques et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Exposure to adversity during the first years of life might already be biologically embedded well before adult life. Thus, the impact of different stressful experiences needs to be explored. This study aims to examine if the association between being victimized (adverse childhood experiences-ACEs and bullying) and (hs-) C-Reactive Protein (CRP) is explained by the influence of adversity on the body mass index (BMI) of the child. We included children from the Portuguese birth cohort Generation XXI (n = 3712) that at 10 years of age completed a questionnaire on the exposure to ACEs and bullying victimization, assessed by an adaptation from the original ACEs study and an adaptation of The Bully Scale Survey, respectively. Following an overnight fast, a venous blood sample was collected by trained nurses and hs-CRP was assayed in fresh blood samples. Weight and height were measured with the child in underwear and bare feet. Weight was measured to the nearest one-tenth of a kilogram with the use of a digital scale (Tanita), and height was measured to the nearest one-tenth of a centimetre with the use of a wall stadiometer (seca®). BMI was calculated as the value of weight (kg) over squared height (m), and computed as an age- and sex-specific BMI standard deviation (SD) score (z-score), according to the World Health Organization Child Growth Standards (5-19 years). Regression coefficients and respective 95% Confidence Interval [β(95%CI)] were computed using path analysis. We observed that ACEs had a positive total effect on hs-CRP at the age of 10 years (β = 0.06; 95%CI: -0.02; 0.15). A direct effect (β = 0.02; 95%CI: -0.01; 0.06) accounted for 66.1% of the association between ACEs and hs-CRP. A positive total effect of bullying victimization on hs-CRP (β = 0.20; 95%CI: 0.06; 0.34) was observed. A direct effect (β = 0.08; 95%CI: -0.05; 0.21) accounted for 40.0% of the association, while an indirect effect through BMI (β = 0.12; 95%CI: 0.06; 0.18) explained 60.0% of the pathway between bullying victimization and hs-CRP. Results suggest that there might be different mechanisms involved in the biological embedding of childhood experiences. BMI seems to explain a great part of the association between exposure to bullying victimization and hs-CRP at 10 years of age. Further research is still needed to better understand the mechanisms explaining the emergence and persistence of health poorer outcomes later in life for victims of abuse., Competing Interests: The authors have declared that no competing interests exist.

Peptide-based vaccines have demonstrated to be an important way to induce long-lived immune responses and, therefore, a promising strategy in the rational of vaccine development. As to malaria, among the classic vaccine targets, the Apical membrane antigen (AMA-1) was proven to have important B cell epitopes that can induce specific immune response and, hence, became key players for a vaccine approach. The peptides selection was carried out using a bioinformatic approach based on Hidden Markov Models profiles of known antigens and propensity scale methods based on hydrophilicity and secondary structure prediction. The antigenicity of the selected B-cell peptides was assessed by multiple serological assays using sera from acute P.vivax infected subjects. The synthetic peptides were recognized by 45.5%, 48.7% and 32.2% of infected subjects for peptides I, II and III respectively. Moreover, when synthetized together (tripeptide), the reactivity increases up to 62%, which is comparable to the reactivity found against the whole protein PvAMA-1 (57%). Furthermore, IgG reactivity against the tripeptide after depletion was reduced by 42%, indicating that these epitopes may be responsible for a considerable part of the protein immunogenicity. These results represent an excellent perspective regarding future chimeric vaccine constructions that may come to contemplate several targets with the potential to generate the robust and protective immune response that a vivax malaria vaccine needs to succeed., Competing Interests: The authors have declared that no competing interests exist.

The efficacy and non-target arthropod effects of transgenic DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 Bt cotton, expressing proteins Cry1Ac, Cry1F and Vip3Aa19, was examined through field trials in Brazil. Fifteen field efficacy experiments were conducted from 2014 through the 2020 growing season across six different states in Brazil to evaluate performance against key lepidopteran pests through artificial infestations of Chrysodeixis includens (Walker), Spodoptera frugiperda (J.E. Smith,1797), Spodoptera cosmioides (Walker, 1858) and Chloridea virescens (F., 1781), and natural infestations of Alabama argillacea (Hübner) and S. frugiperda. The impact of this Bt cotton technology on the non-target arthropod community in Brazilian cotton production systems was also assessed in a multi-site experiment. DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton significantly reduced the feeding damage caused by S. frugiperda, S. cosmioides, C. includens, C. virescens and A. argillacea, causing high levels of mortality (greater than 99%) to all target lepidopteran pests evaluated during vegetative and/or reproductive stages of crop development. Non-target arthropod community-level analyses confirmed no unintended effects on the arthropod groups monitored. These results demonstrate the value of transgenic Bt cotton containing event DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 for consideration as part of an integrated approach for managing key lepidopteran pests in Brazilian cotton production systems., Competing Interests: The authors have declared that no competing interests exist. Corteva Agriscience provided support in the form of salaries for authors Luiz H. Marques, Miles D. Lepping, Boris A. Castro, Antonio C. Santos, Jaedino Rossetto, Marcelo Z. Nunes, Oscar A. B. N. Silva, Verissimo G. M. de Sá, Valeria F. Moscardini, Timothy Nowatzki, Mark L. Dahmer, but did not have any additional role in the conduction of field trials, data collection and analysis. The specific role of these authors are articulated in the ’author contributions’ section. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: The Self-Administered Food Security Survey Module for Children was developed to assess food insecurity of individual children and has not been used in Portugal. We aimed to apply the mixture item response theory model to the Self-Administered Food Security Survey Module for Children, to assess its reliability and validity, and to estimate the cut-offs of the food security status for Portuguese children., Methods: The scale was self-administered to 2132 children of the Generation XXI birth cohort. The internal consistency was assessed using Cronbach's alpha. We evaluated dimensionality and/or clustering, and Latent Class Analysis, Latent Trait Analysis and Mixture Latent Trait Analysis were tested. The number of classes and/or traits were defined according to the Akaike Information Criterion, Bayesian Information Criterion, Adjusted Bayesian Information Criterion, Vuong-Lo-Mendell-Rubin Likelihood Ratio Test, Bootstrapped Likelihood Ratio Test and Entropy. Construct validity was explored using socio-demographic characteristics. The classification tree was used to define cut-offs to predict cluster membership., Results: The best model was a Mixture Latent Trait Analysis with 1 factor and 2 classes (food security and food insecurity), assuming class variant item parameters (for items 1 and 3). Based on the estimated posterior probabilities, the food insecurity prevalence was 17.6%. Cronbach's alpha was 0.617. A higher proportion of less-educated mothers and low-income households was observed in the food insecurity class. The classification tree showed an accuracy of 100.0% by identifying the food security and food insecurity groups., Conclusion: Our results supported that the Self-Administered Food Security Survey Module for Children provides a valid and reliable measure, which allows the identification of food insecurity among Portuguese children., Competing Interests: The authors have declared that no competing interests exist.

Purpose: We aimed to describe bone formation and resorption markers in generally healthy prepubertal children using total alkaline phosphatase (tALP), osteocalcin (OC) and β-isomerized C-terminal telopeptides of type I collagen (β-CTx) serum concentrations and to estimate markers' correlations with anthropometric growth (height, weight, body mass index and trajectories of weight gain) as well as bone mineral content (BMC) and areal density (aBMD)., Methods: We assessed 395 7-year-old children from the Generation XXI cohort with tALP, OC and β-CTx concentrations determined from a fasting venous blood sample and BMC/aBMD measured by dual-energy X-ray absorptiometry. Gender-specific reference intervals for tALP, OC and β-CTx in 7-year-old children were established by calculating the 2.5th and 97.5th percentiles. Pearson and partial correlation coefficients (controlling for sex, age, body size and season) between bone markers and growth measures were computed., Results: tALP increased with height (rpartial controlled for sex = 0.26, 95%CI: 0.17, 0.35), was higher in overweight than in healthy weight children, and in children who gained weight above average during infancy. No correlations were found between OC or β-CTx and growth. In girls, OC was slightly correlated with subtotal BMC (rpartial = 0.22, 95%CI: 0.08, 0.35), subtotal aBMD (rpartial = 0.20, 95%CI: 0.06, 0.33) and lumbar spine aBMD (rpartial = 0.23, 95%CI: 0.09, 0.36). tALP and β-CTx were not correlated with any of the DXA-derived bone measures., Conclusion: This study contributed to the description of bone turnover at 7 years of age and suggested that bone metabolism markers measured in a single point in time have limited ability to describe anthropometric growth and overall bone status in generally healthy prepubertal children., Competing Interests: The authors have declared that no competing interests exist.

Twin births are an important public health issue due to health complications for both mother and children. While it is known that contemporary factors have drastically changed the epidemiology of twins in certain developed countries, in Brazil, relevant data are still scarce. Thus, we carried out a population-based study of live births in spatial and temporal dimensions using data from Brazil's Live Birth Information System, which covers the entire country. Over 41 million births registered between 2001 and 2014 were classified as singleton, twin or multiple. Twinning rates (TR) averaged 9.41 per 1,000 for the study period and a first-order autoregressive model of time-series analysis revealed a global upward trend over time; however, there were important regional differences. In fact, a Cluster and Outlier Analysis (Anselin Local Moran's I) was performed and identified clusters of high TR in an area stretching from the south of Brazil's Northeast Region to the South Region (Global Moran Index = 0.062, P < 0.001). Spearman's correlation coefficient and a Wilcoxon matched pairs test revealed a positive association between Human Development Index (HDI) and TRs in different scenarios, suggesting that the HDI might be an important indicator of childbearing age and assisted reproduction techniques in Brazil. Furthermore, there was a sharp increase of 26.42% in TR in women aged 45 and over during study period. The upward temporal trend in TRs is in line with recent observations from other countries, while the spatial analysis has revealed two very different realities within the same country. Our approach to TR using HDI as a proxy for underlying socioeconomic changes can be applied to other developing countries with regional inequalities resembling those found in Brazil., Competing Interests: The authors have declared that no competing interests exist.

Field-scale studies that examine the potential for adverse effects of Bt crop technology on non-target arthropods may supplement data from laboratory studies to support an environmental risk assessment. A three year field study was conducted in Brazil to evaluate potential for adverse effects of cultivating soybean event DAS-81419-2 that produces the Cry1Ac and Cry1F proteins. To do so, we examined the diversity and abundance of non-target arthropods (NTAs) in Bt soybean in comparison with its non-Bt near isoline, with and without conventional insecticide applications, in three Brazilian soybean producing regions. Non-target arthropod abundance was surveyed using Moericke traps (yellow pan) and pitfall trapping. Total abundance (N), richness (S), Shannon-Wiener (H'), Simpson's (D) and Pielou's evenness (J) values for arthropod samples were calculated for each treatment and sampling period (soybean growth stages). A faunistic analysis was used to select the most representative NTAs which were used to describe the NTA community structure associated with soybean, and to test for effects due to the treatments effects via application of the Principal Response Curve (PRC) method. Across all years and sites, a total of 254,054 individuals from 190 taxa were collected by Moericke traps, while 29,813 individuals from 100 taxa were collected using pitfall traps. Across sites and sampling dates, the abundance and diversity measurements of representative NTAs were not significantly affected by Bt soybean as compared with non-sprayed non-Bt soybean. Similarly, community analyses and repeated measures ANOVA, when applicable, indicated that neither Bt soybean nor insecticide sprays altered the structure of the NTA communities under study. These results support the conclusion that transgenic soybean event DAS-81419-2 producing Cry1Ac and Cry1F toxins does not adversely affect the NTA community associated with soybean.

Objectives: The contribution of adiposity to cardiovascular and diabetes risk justifies the inclusion of an adiposity measure, usually waist circumference, in the definition of metabolic syndrome. However, waist circumference thresholds differ across populations. Our aim was to assess which adiposity measure performs the best in identifying the metabolic syndrome in a sample of Portuguese participants and to estimate cut-off values for these measures., Methods: Data were obtained from a cross-sectional study (PORMETS study) conducted in Portugal between 2007 and 2009. A representative sample of non-institutionalized adults, comprising 3,956 participants, aged 18 years and older, was evaluated. A structured questionnaire was administered, collecting information on personal medical history, socio-demographics and behavioral characteristics. Anthropometrics, blood pressure and venous blood samples were also obtained. Metabolic syndrome was defined according to the Joint Interim Statement of the International Diabetes Federation Task Force on Epidemiology recommended criteria. Elevated cardiometabolic risk was considered when two or more of the four criteria of metabolic syndrome were present, excluding the waist circumference component. A receiver operating characteristic curve was used to estimate cut-off points., Results: This study found that waist-to-height ratio, waist circumference and body adiposity index performed better than other adiposity measures, such as body mass index. The estimated cut-off points for waist-to-height ratio, waist circumference and body adiposity index in women and men were 0.564 / 89 cm / 27.4 and 0.571 / 93.5 cm / 25.5, respectively., Conclusion: As waist circumference is currently used as the adiposity measure in the definition of metabolic syndrome and as no relevant differences were observed between this measure and waist-to-height ratio, it is likely that no modification to the metabolic syndrome definition needs to be proposed. Moreover, this study also confirmed the applicability of European cut-off points in the Portuguese population.

It has been proposed that copy number variations (CNVs) are associated with increased risk of autism spectrum disorder (ASD) and, in conjunction with other genetic changes, contribute to the heterogeneity of ASD phenotypes. Array comparative genomic hybridization (aCGH) and exome sequencing, together with systems genetics and network analyses, are being used as tools for the study of complex disorders of unknown etiology, especially those characterized by significant genetic and phenotypic heterogeneity. Therefore, to characterize the complex genotype-phenotype relationship, we performed aCGH and sequenced the exomes of two affected siblings with ASD symptoms, dysmorphic features, and intellectual disability, searching for de novo CNVs, as well as for de novo and rare inherited point variations-single nucleotide variants (SNVs) or small insertions and deletions (indels)-with probable functional impacts. With aCGH, we identified, in both siblings, a duplication in the 4p16.3 region and a deletion at 8p23.3, inherited by a paternal balanced translocation, t(4, 8) (p16; p23). Exome variant analysis found a total of 316 variants, of which 102 were shared by both siblings, 128 were in the male sibling exome data, and 86 were in the female exome data. Our integrative network analysis showed that the siblings' shared translocation could explain their similar syndromic phenotype, including overgrowth, macrocephaly, and intellectual disability. However, exome data aggregate genes to those already connected from their translocation, which are important to the robustness of the network and contribute to the understanding of the broader spectrum of psychiatric symptoms. This study shows the importance of using an integrative approach to explore genotype-phenotype variability., Competing Interests: The authors have declared that no competing interests exist.

Although previous studies have described opposing states in upwelling regions, i.e., the rise of cold nutrient-rich waters and prevalence of surface warm nutrient-poor waters, few have addressed the transition from one state to the other. This study aimed to describe the microbial and viral structure during this transition and was able to obtain the taxonomic and metabolic compositions as well as physical-chemical data. This integrated approach allowed for a better understanding of the dynamics of the downwelling upwelling transition, suggesting that a wealth of metabolic processes and ecological interactions are occurring in the minute fractions of the plankton (femto, pico, nano). These processes and interactions included evidence of microbial predominance during downwelling (with nitrogen recycling and aerobic anoxygenic photosynthesis), different viral predation pressures over primary production in different states (cyanobacteria vs eukaryotes), and a predominance of diatoms and selected bacterial and archaeal groups during upwelling (with the occurrence of a wealth of nitrogen metabolism involving ammonia). Thus, the results provided insights into which microbes, viruses and microbial-mediated processes are probably important in the functioning of upwelling systems.

Objective: To evaluate the association between obesity indices and blood pressure (BP) at 4 years of age, in each sex, and to quantify to which extent this association is mediated by inflammation and insulin resistance (IR)., Materials and Methods: We studied 1250 4-year-old children selected from the population-based birth cohort Generation XXI. Associations between body mass index (BMI) z-score and waist-to-height ratio (WHtR), office BP, inflammation (high sensitivity C-reactive protein) and IR (HOMA-IR index) were assessed. Path Analysis, a modified multivariate regression approach, was applied to test causal models and quantify direct and indirect effects of predictors of systolic (SBP) and diastolic BP (DBP)., Results: SBP and DBP increased significantly with BMI and WHtR in both sexes. There was a strong direct association (explaining 74.1-93.2% of the total association) of both measures of adiposity with SBP, in both sexes. This association was additionally indirectly mediated by IR, particularly regarding WHtR (20.5% in girls and 9.4% in boys). Mediation by inflammation did not reach statistical significance in either sex. Regarding DBP, the direct effect of adiposity was strong (>95% for BMI and WHtR in boys) and the mediation by IR was much smaller in boys than in girls., Discussion: The direct association between adiposity and BP in healthy 4-year-old children is strong and IR plays an important mediating role. The strength of effects of IR and inflammation suggests sex differences in the complex interplay between BP, adiposity and inflammation.

Influenza viruses pose a serious global health threat, particularly in light of newly emerging strains, such as the avian influenza H5N1 and H7N9 viruses. Vaccination remains the primary method for preventing acquiring influenza or for avoiding developing serious complications related to the disease. Vaccinations based on inactivated split virus vaccines or on chemically inactivated whole virus have some important drawbacks, including changes in the immunogenic properties of the virus. To induce a greater mucosal immune response, intranasally administered vaccines are highly desired as they not only prevent disease but can also block the infection at its primary site. To avoid these drawbacks, hydrostatic pressure has been used as a potential method for viral inactivation and vaccine production. In this study, we show that hydrostatic pressure inactivates the avian influenza A H3N8 virus, while still maintaining hemagglutinin and neuraminidase functionalities. Challenged vaccinated animals showed no disease signs (ruffled fur, lethargy, weight loss, and huddling). Similarly, these animals showed less Evans Blue dye leakage and lower cell counts in their bronchoalveolar lavage fluid compared with the challenged non-vaccinated group. We found that the whole inactivated particles were capable of generating a neutralizing antibody response in serum, and IgA was also found in nasal mucosa and feces. After the vaccination and challenge we observed Th1/Th2 cytokine secretion with a prevalence of IFN-γ. Our data indicate that the animals present a satisfactory immune response after vaccination and are protected against infection. Our results may pave the way for the development of a novel pressure-based vaccine against influenza virus.

In previous work, a new lipase and its cognate foldase were identified and isolated from a metagenomic library constructed from soil samples contaminated with fat. This new lipase, called LipG9, is a true lipase that shows specific activities that are comparable to those of well-known industrially-used lipases with high activity against long-chain triglycerides. In the present work, LipG9 was co-expressed and co-immobilized with its foldase, on an inert hydrophobic support (Accurel MP1000). We studied the performance of this immobilized LipG9 (Im-LipG9) in organic media, in order to evaluate its potential for use in biocatalysis. Im-LipG9 showed good stability, maintaining a residual activity of more than 70% at 50 °C after incubation in n-heptane (log P 4.0) for 8 h. It was also stable in polar organic solvents such as ethanol (log P -0.23) and acetone (log P -0.31), maintaining more than 80% of its original activity after 8 h incubation at 30 °C. The synthesis of ethyl esters was tested with fatty acids of different chain lengths in n-heptane at 30 °C. The best conversions (90% in 3 h) were obtained for medium and long chain saturated fatty acids (C8, C14 and C16), with the maximum specific activity, 29 U per gram of immobilized preparation, being obtained with palmitic acid (C16). Im-LipG9 was sn-1,3-specific. In the transesterification of the alcohol (R,S)-1-phenylethanol with vinyl acetate and the hydrolysis of the analogous ester, (R,S)-1-phenylethyl acetate, Im-LipG9 showed excellent enantioselectivity for the R-isomer of both substrates (E> 200), giving an enantiomeric excess (ee) of higher than 95% for the products at 49% conversion. The results obtained in this work provide the basis for the development of applications of LipG9 in biocatalysis.

The experiences induced by psychedelics share a wide variety of subjective features, related to the complex changes in perception and cognition induced by this class of drugs. A remarkable increase in introspection is at the core of these altered states of consciousness. Self-oriented mental activity has been consistently linked to the Default Mode Network (DMN), a set of brain regions more active during rest than during the execution of a goal-directed task. Here we used fMRI technique to inspect the DMN during the psychedelic state induced by Ayahuasca in ten experienced subjects. Ayahuasca is a potion traditionally used by Amazonian Amerindians composed by a mixture of compounds that increase monoaminergic transmission. In particular, we examined whether Ayahuasca changes the activity and connectivity of the DMN and the connection between the DMN and the task-positive network (TPN). Ayahuasca caused a significant decrease in activity through most parts of the DMN, including its most consistent hubs: the Posterior Cingulate Cortex (PCC)/Precuneus and the medial Prefrontal Cortex (mPFC). Functional connectivity within the PCC/Precuneus decreased after Ayahuasca intake. No significant change was observed in the DMN-TPN orthogonality. Altogether, our results support the notion that the altered state of consciousness induced by Ayahuasca, like those induced by psilocybin (another serotonergic psychedelic), meditation and sleep, is linked to the modulation of the activity and the connectivity of the DMN.

Background: At the triple border Brazil/Paraguay/Argentina there is easy mobility from one city to another for economic and tourism activities. This constant and fast population mobility is mainly to visit Iguazu Falls, in the Iguazu River, on the border of the Brazilian state of Paraná and the Argentina. As the incidence of tuberculosis is high in this setting, our study aimed to establish a first baseline of circulating genotypic lineages of Mycobacterium tuberculosis., Methodology/principal Findings: This study included 120 patients from 10 cities in southwestern Paraná, Brazil with pulmonary symptoms, from July 2009 to July 2011. Information about sex, age, clinical features and address was collected by reviewing the national tuberculosis notification database. Of these, 96 (80%) isolates were identified as M. tuberculosis and 22 (22.9%) were drug resistant (20, 20.8% INH mono-resistant and 2, 2.1% multidrug-resistant). All isolates were subjected to genotyping by Spoligotyping and MIRU-VNTR typing. The distribution of the isolates analyzed by spoligotyping revealed 30 distinct patterns. The four mainly detected clades were Latin American and Mediterranean (LAM), ill-defined T, Haarlem (H) and S. The MIRU-VNTR showed 85 distinct patterns. Spoligotyping combined to MIRU-VNTR allowed 90 distinct patterns., Conclusions/significance: Our study demonstrated that there is significant molecular diversity in circulating M. tuberculosis, with predominance of the LAM and T clades in cities of southwestern Paraná, Brazil, bordering Argentina and Paraguay.

Alzheimer's disease (AD) is characterized by progressive cognitive decline associated with a featured neuropathology (neuritic plaques and neurofibrillary tangles). Several studies have implicated oxidative damage to DNA, DNA repair, and altered cell-cycle regulation in addition to cell death in AD post-mitotic neurons. However, there is a lack of studies that systematically assess those biological processes in patients with AD neuropathology but with no evidence of cognitive impairment. We evaluated markers of oxidative DNA damage (8-OHdG, H2AX), DNA repair (p53, BRCA1, PTEN), and cell-cycle (Cdk1, Cdk4, Cdk5, Cyclin B1, Cyclin D1, p27Kip1, phospho-Rb and E2F1) through immunohistochemistry and cell death through TUNEL in autopsy hippocampal tissue samples arrayed in a tissue microarray (TMA) composed of three groups: I) "clinical-pathological AD" (CP-AD)--subjects with neuropathological AD (Braak ≥ IV and CERAD = B or C) and clinical dementia (CDR ≥ 2, IQCODE>3.8); II) "pathological AD" (P-AD)--subjects with neuropathological AD (Braak ≥ IV and CERAD = B or C) and without cognitive impairment (CDR 0, IQCODE<3.2); and III) "normal aging" (N)--subjects without neuropathological AD (Braak ≤ II and CERAD 0 or A) and with normal cognitive function (CDR 0, IQCODE<3.2). Our results show that high levels of oxidative DNA damage are present in all groups. However, significant reductions in DNA repair and cell-cycle inhibition markers and increases in cell-cycle progression and cell death markers in subjects with CP-AD were detected when compared to both P-AD and N groups, whereas there were no significant differences in the studied markers between P-AD individuals and N subjects. This study indicates that, even in the setting of pathological AD, healthy cognition may be associated with a preserved repair to DNA damage, cell-cycle regulation, and cell death in post-mitotic neurons.

Several probiotic bacteria have been proposed for treatment or prevention of inflammatory bowel diseases (IBD), showing a protective effect in animal models of experimental colitis and for some of them also in human clinical trials. While most of these probiotic bacteria are isolated from the digestive tract, we recently reported that a Lactobacillus strain isolated from cheese, L. delbrueckii subsp. lactis CNRZ327 (Lb CNRZ327), also possesses anti-inflammatory effects in vitro and in vivo, demonstrating that common dairy bacteria may be useful in the treatment or prevention of IBD. Here, we studied the mechanisms underlying the protective effects of Lb CNRZ327 in vivo, in a mouse dextran sodium sulfate (DSS) colitis model. During colitis, Lb CNRZ327 modulated the production of TGF-β, IL-6, and IL-12 in colonic tissue and of TGF-β and IL-6 in the spleen, and caused an expansion of CD4+Foxp3+ regulatory T cells in the cecal lymph nodes. Moreover, a strong tendency to CD4+Foxp3+ expansion was also observed in the spleen. The results of this study for the first time show that orally administered dairy lactobacilli can not only modulate mucosal but also systemic immune responses and constitute an effective treatment of IBD.

Background: Where neurocysticercosis (NCC) is endemic, chronic calcified neurocysticercosis (cNCC) can be observed in patients with mesial temporal lobe epilepsy associated with hippocampal sclerosis (MTLE-HS). Considering that both disorders cause recurrent seizures or cognitive impairment, we evaluated if temporal lobectomy is cognitively safe and effective for seizure control in MTLE-HS plus cNCC., Methods: Retrospective cohort study of neuropsychological profile and surgical outcome of 324 MTLE-HS patients submitted to temporal lobectomy, comparing the results according to the presence or absence of cNCC., Findings: cNCC occurred in 126 (38.9%) of our MTLE-HS patients, a frequency higher than expected, more frequently in women than in men (O.R. = 1.66; 95% C.I. = 1.05-2.61; p = 0.03). Left-side (but not right side) surgery caused impairment in selected neuropsychological tests, but this impairment was not accentuated by the presence of cNCC. Ninety-four (74.6%) patients with MTLE-HS plus cNCC and 153 patients (77.3%) with MTLE-HS alone were Engel class I after surgery (O.R. = 1.16; 95% C.I. = 0.69-1.95; p = 0.58). However, the chances of Engel class IA were significantly lower in MTLE-HS plus cNCC than in patients with MTLE-HS alone (31.7% versus 48.5%; O.R. = 2.02; 95% C.I. = 1.27-3.23; p = 0.003). Patients with MTLE-HS plus cNCC showed higher rates of Engel class ID (15.1% versus 6.6%; O.R. = 2.50; 95% C.I. = 1.20-5.32; p = 0.012)., Interpretation: cNCC can be highly prevalent among MTLE-HS patients living in areas where neurocysticercosis is endemic, suggesting a cause-effect relationship between the two diseases. cNCC does not add further risk for cognitive decline after surgery in MTLE-HS patients. The rates of Engel class I outcome were very similar for the two groups; however, MTLE-HS plus cNCC patients achieved Engel IA status less frequently, and Engel ID status more frequently. Temporal lobectomy can be safely performed in most patients with MTLE-HS plus cNCC without affecting cognitive outcome. Long-term surgical seizure control in MTLE-HS plus cNCC is still satisfactory, as long as selected patients remain under medication.

Background: Neuromyelitis optica (NMO) is considered relatively more common in non-Whites, whereas multiple sclerosis (MS) presents a high prevalence rate, particularly in Whites from Western countries populations. However, no study has used ancestry informative markers (AIMs) to estimate the genetic ancestry contribution to NMO patients., Methods: Twelve AIMs were selected based on the large allele frequency differences among European, African, and Amerindian populations, in order to investigate the genetic contribution of each ancestral group in 236 patients with MS and NMO, diagnosed using the McDonald and Wingerchuck criteria, respectively. All 128 MS patients were recruited at the Faculty of Medicine of Ribeirão Preto (MS-RP), Southeastern Brazil, as well as 108 healthy bone marrow donors considered as healthy controls. A total of 108 NMO patients were recruited from five Neurology centers from different Brazilian regions, including Ribeirão Preto (NMO-RP)., Principal Findings: European ancestry contribution was higher in MS-RP than in NMO-RP (78.5% vs. 68.7%) patients. In contrast, African ancestry estimates were higher in NMO-RP than in MS-RP (20.5% vs. 12.5%) patients. Moreover, principal component analyses showed that groups of NMO patients from different Brazilian regions were clustered close to the European ancestral population., Conclusions: Our findings demonstrate that European genetic contribution predominates in NMO and MS patients from Brazil.

Background: Botulinum toxin type A (btxA) is one of the main treatment choices for patients with spasticity. Prosigne® a new released botulinum toxin serotype A may have the same effectiveness as Botox® in focal dystonia. However, there are no randomized clinical trials comparing these formulations in spasticity treatment. The aim of our study was to compare the efficacy and safety of Prosigne® with Botox® in the treatment of spasticity., Methodology/principal Findings: We performed a double-blind, randomized, crossover study consisting of 57 patients with clinically meaningful spasticity. The patients were assessed at baseline, 4 and 12 weeks after Prosigne® or Botox® administration. The main outcomes were changes in the patients' Modified Ashworth Scale (MAS), Functional Independence Measure (FIM) and Pediatric Evaluation of Disability Inventory (PEDI) scores and adverse effects related to the botulinum toxin. Both of the toxins were significantly effective in relieving the level of spasticity in adults and children. There were no significant differences found between the Prosigne® and Botox® treatments regarding their MAS, FIM and PEDI scores. Likewise, the incidence of adverse effects was similar between the two groups., Conclusion: Our results suggest that Prosigne® and Botox® are both efficient and comparable with respect to their efficacy and safety for the three month treatment of spasticity., Trial Registration: ClinicalTrials.gov NCT00819065.

Background: Epithelia act as physical barriers protecting living organisms and their organs from the surrounding environment. Simple epithelial tissues have the capacity to efficiently repair wounds through a resealing mechanism. The known molecular mechanisms underlying this process appear to be conserved in both vertebrates and invertebrates, namely the involvement of the transcription factors Grainy head (Grh) and Fos. In Drosophila, Grh and Fos lead to the activation of wound response genes required for epithelial repair. ERK is upstream of this pathway and known to be one of the first kinases to be activated upon wounding. However, it is still unclear how ERK activation contributes to a proper wound response and which molecular mechanisms regulate its activation., Methodology/principal Findings: In a previous screen, we isolated mutants with defects in wound healing. Here, we describe the role of one of these genes, hole-in-one (holn1), in the wound healing process. Holn1 is a GYF domain containing protein that we found to be required for the activation of several Grh and Fos regulated wound response genes at the wound site. We also provide evidence suggesting that Holn1 may be involved in the Ras/ERK signaling pathway, by acting downstream of ERK. Finally, we show that wound healing requires the function of EGFR and ERK signaling., Conclusions/significance: Based on these data, we conclude that holn1 is a novel gene required for a proper wound healing response. We further propose and discuss a model whereby Holn1 acts downstream of EGFR and ERK signaling in the Grh/Fos mediated wound closure pathway.

Background: Secondary bone marrow (BM) myelodysplastic syndromes (MDS) are increasingly common, as a result of radio or chemotherapy administered to a majority of cancer patients. Patients with secondary MDS have increased BM cell apoptosis, which results in BM dysfunction (cytopenias), and an increased risk of developing fatal acute leukemias. In the present study we asked whether TNF-alpha, known to regulate cell apoptosis, could modulate the onset of secondary MDS., Principal Findings: We show that TNF-alpha is induced by irradiation and regulates BM cells apoptosis in vitro and in vivo. In contrast to irradiated wild type (WT) mice, TNF-alpha deficient (TNF-alpha KO) mice or WT mice treated with a TNF-alpha-neutralizing antibody were partially protected from the apoptotic effects of irradiation. Next we established a 3-cycle irradiation protocol, in which mice were sub-lethally irradiated once monthly over a 3 month period. In this model, irradiated WT mice presented loss of microsatellite markers on BM cells, low white blood cell (WBC) counts, reduced megakaryocyte (MK) and platelet levels (thrombocytopenia) and macrocytic anemia, phenoypes that suggest the irradiation protocol resulted in BM dysfunction with clinical features of MDS. In contrast, TNF-alpha KO mice were protected from the irradiation effects: BM cell apoptosis following irradiation was significantly reduced, concomitant with sustained BM MK numbers and absence of other cytopenias. Moreover, irradiated WT mice with long term (> or = 5 months) BM dysfunction had increased BM angiogenesis, MMPs and VEGF and NFkB p65, suggestive of disease progression., Conclusion: Taken together, our data shows that TNF-alpha induction following irradiation modulates BM cell apoptosis and is a crucial event in BM dysfunction, secondary MDS onset and progression.