Search

Infection with Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis (TB), remains a significant global epidemic. Host resistance to Mtb depends on both adaptive and innate immunity mechanisms, including development of antigen-specific CD4 and CD8 T cells, production of inflammatory cytokines, bacterial phagocytosis and destruction within phagolysosomes, host cell apoptosis, and autophagy. A key regulatory mechanism in innate immunity is the attachment of the small protein ubiquitin to protein and lipid targets by the enzymatic activity of ubiquitin ligases. Here, we summarize the latest advances on the role of ubiquitination and ubiquitin ligases in host immunity against Mtb, with a focus on innate immunity signaling, inflammation, and antimicrobial autophagy. Understanding how ubiquitin ligases mediate immunity to Mtb, and the specific substrates of distinct ubiquitin ligases in the context of Mtb infection, could facilitate development of new host-directed antimicrobials.

IntroductionThe tumor microenvironment (TME) of glioblastoma (GB) is characterized by an increased infiltration of immunosuppressive cells that attenuate the antitumor immune response. The participation of neutrophils in tumor progression is still controversial and a dual role in the TME has been proposed. In this study, we show that neutrophils are reprogrammed by the tumor to ultimately promote GB progression.MethodsUsing in vitro and in vivo assays, we demonstrate the existence of bidirectional GB and neutrophil communication, directly promoting an immunosuppressive TME. Results and discussionNeutrophils have shown to play an important role in tumor malignancy especially in advanced 3D tumor model and Balb/c nude mice experiments, implying a time- and neutrophil concentration-dependent modulation. Studying the tumor energetic metabolism indicated a mitochondria mismatch shaping the TME secretome. The given data suggests a cytokine milieu in patients with GB that favors the recruitment of neutrophils, sustaining an anti-inflammatory profile which is associated with poor prognosis. Besides, glioma-neutrophil crosstalk has sustained a tumor prolonged activation via NETs formation, indicating the role of NFκB signaling in tumor progression. Moreover, clinical samples have indicated that neutrophil-lymphocyte ratio (NLR), IL-1β, and IL-10 are associated with poor outcomes in patients with GB. ConclusionThese results are relevant for understanding how tumor progression occurs and how immune cells can help in this process.

The wine industry produces significant amounts of by-products and residues that are not properly managed, posing an environmental problem. Grape must surplus, vine shoots, and wine lees have the potential to be used as renewable resources for the production of energy and chemicals. Metabolic engineering efforts have established Saccharomyces cerevisiae as an efficient microbial cell factory for biorefineries. Current biorefineries designed for producing multiple products often rely on just one feedstock, but the bioeconomy would clearly benefit if these biorefineries could efficiently convert multiple feedstocks. Moreover, to reduce the environmental impact of fossil fuel consumption and maximize production economics, a biorefinery should be capable to supplement the manufacture of biofuel with the production of high-value products. This study proposes an integrated approach for the valorization of diverse wastes resulting from winemaking processes through the biosynthesis of xylitol and ethanol. Using genetically modified S. cerevisiae strains, the xylose-rich hemicellulosic fraction of hydrothermally pretreated vine shoots was converted into xylitol, and the cellulosic fraction was used to produce bioethanol. In addition, grape must, enriched in sugars, was efficiently used as a low-cost source for yeast propagation. The production of xylitol was optimized, in a Simultaneous Saccharification and Fermentation process configuration, by adjusting the inoculum size and enzyme loading. Furthermore, a yeast strain displaying cellulases in the cell surface was applied for the production of bioethanol from the glucan-rich cellulosic. With the addition of grape must and/or wine lees, high ethanol concentrations were reached, which are crucial for the economic feasibility of distillation. This integrated multi-feedstock valorization provides a synergistic alternative for converting a range of winery wastes and by-products into biofuel and an added-value chemical while decreasing waste released to the environment., This study was supported by the Portuguese Foundation for Science and Technology (FCT, Portugal) under the scope of the strategic funding of UIDB/04469/2020, the PhD grant (SFRH/BD/132717/2017 to SLB) and BIOVINO project (0688_BIOVINO_6_E) funded by INTERREG Espana - Portugal and European Regional. A.R. acknowledges the Ministry of Science and Innovation (MICIN) through the grant RYC2020- 030690-I., info:eu-repo/semantics/publishedVersion

In this paper, the design and development of a novel Fast-Field Cycling (FFC) Nuclear Magnetic Resonance (NMR) relaxometer’s electromagnet is described. This magnet is tailored to increase the relaxometers’s usability, by increasing its portability capacities. It presents a compact toroidal shaped iron core, allowing to operate in a field range of 0 to 0.21 T, with high field homogeneity (less than 800 ppm in a volume of ≈ 0.57 cm3 ), low power consumption and reduced losses (about 40W). The simulation software COMSOL Multiphysics® is used to characterize the induced magnetic field, the heating and the cooling effects. The proposed optimized layout constitutes an innovative solution for FFC magnets.

BackgroundConsidering the potential role of miRNAs as biomarkers and their interaction with both nuclear and mitochondrial genes, we investigated the miRNA expression profile in type 1 diabetes (T1DM) patients, including the pathways in which they are involved considering both nuclear and mitochondrial functions.MethodsWe analyzed samples of T1DM patients and control individuals (normal glucose tolerance) by high throughput miRNA sequencing (miRNome). Next, five miRNAs – hsa-miR-26b-5p, hsa-let-7i-5p, hsa-miR-143-3p, hsa-miR-501-3p and hsa-miR-100-5p – were validated by RT-qPCR. The identification of target genes was extracted from miRTarBase and mitoXplorer database. We also performed receiver operating characteristic (ROC) curves and miRNAs that had an AUC > 0.85 were considered potential biomarkers.ResultsOverall, 41 miRNAs were differentially expressed in T1DM patients compared to control. Hsa-miR-21-5p had the highest number of predicted target genes and was associated with several pathways, including insulin signaling and apoptosis. 34.1% (14/41) of the differentially expressed miRNAs also targeted mitochondrial genes, and 80.5% (33/41) of them targeted nuclear genes involved in the mitochondrial metabolism. All five validated miRNAs were upregulated in T1DM. Among them, hsa-miR-26b-5p showed AUC>0.85, being suggested as potential biomarker to T1DM.ConclusionOur results demonstrated 41 DE miRNAs that had a great accuracy in discriminating T1DM and control group. Furthermore, we demonstrate the influence of these miRNAs on numerous metabolic pathways, including mitochondrial metabolism. Hsa-miR-26b-5p and hsa-miR-21-5p were highlighted in our results, possibly acting on nuclear and mitochondrial dysfunction and, subsequently, T1DM dysregulation.

Kite flying is a popular hobby and sport for children and adults. Despite being illegal in Rio de Janeiro state, Brazil, the use of abrasive threads remains widespread and poses a health risk to both humans and animals. In this study, we analysed the records of 462 magnificent frigatebirds (Fregata magnificens) submitted to rehabilitation centres or found dead along the southern coast of Rio de Janeiro state from October 2016 to August 2020. Of these, 244 individuals (52.8%) presented wing lesions consistent with kite-string injury, which can have a critical impact on the ability of frigatebirds to fly and will ultimately cause their death. Even when veterinary care is provided, only a small proportion of the individuals (2%) will fully recover the ability to fly in order to be released back to the wild. In 2020, an atypical increase in the number of individuals with kite-string injuries (~1,200% increase compared to other years) was noted in the weeks following the suspension of school activities and commerce in response to the COVID-19 pandemic. The number of frigatebirds with kitestring injuries recorded in a given week was positively correlated with internet searches for kite-related terms, which also peaked during the quarantine period of the COVID-19 pandemic. This illustrates how pandemic events may aggravate existing human-wildlife conflicts, and how preparedness plans need to incorporate measures to help communities cope with boredom and isolation during quarantine in ways that do not negatively impact the welfare and conservation of wildlife.

Data on the prevalence of chronic pulmonary aspergillosis (CPA) in patients with active or cured tuberculosis (TB) are scarce, mainly due to diagnostic difficulties. The diagnosis of CPA is based on pulmonary symptoms and chest computed tomography (CT) scans and is considered confirmed when there is microbiological or serological evidence of Aspergillus spp.To estimate the prevalence of CPA in patients treated or undergoing treatment for PTB, seen in two referral hospitals in Mato Grosso do Sul, Brazil.A total of 193 consecutive patients who were treated or previously treated for pulmonary tuberculosis underwent prospective evaluation: (a) clinical evaluation; (b) chest CT scan; (c) sputum examination-culture for fungi and smears for direct mycology; (d) detection of anti-Aspergillus fumigatus antibodies using an enzyme-linked immunosorbent assay Platelia® test; and (e) anti-Aspergillus spp. antibodies were assessed via a DID test.The global prevalence of CPA was 10.9% (95% confidence interval, 7.2%-16.1%), but it increased with the time of TB diagnosis. The variables independently associated with CPA were previous pulmonary tuberculosis over 4 years ago and haemoptysis. Cavities, pleural thickening and the presence of a fungal ball were the most frequent tomographic findings in patients with CPA.The high prevalence observed and its increase over time suggest the need for continuous surveillance of CPA in patients with active or previous pulmonary tuberculosis and throughout life, with clinical, tomographic and serological evaluations (ELISA) for a timely diagnosis and a better prognosis.

The implementation of cost-effective and sustainable biorefineries to substitute the petroleum-based economy is dependent on coupling the production of bioenergy with high-value chemicals. For this purpose, the US Department of Energy identified a group of key target compounds to be produced from renewable biomass. Among them, 5-hydroxymethylfurfural (HMF) can be obtained by dehydration of the hexoses present in biomass and is an extremely versatile molecule that can be further converted into a wide range of higher value compounds. HMF derivatives include 2,5-bis(hydroxymethyl)furan (BHMF), 5-hydroxymethyl-furan-2-carboxylic acid (HMFCA), 2,5-diformylfuran (DFF), 5-formyl-2-furancarboxylic acid (FFCA) and 2,5-furandicarboxylic acid (FDCA), all presenting valuable applications, in polymers, bioplastics and pharmaceuticals. Biocatalysis conversion of HMF into its derivatives emerges as a green alternative, taking into account the high selectivity of enzymes and the mild reaction conditions used. Considering these factors, this work reviews the use of microorganisms as whole-cell biocatalysts for the production of HMF derivatives. In the last years, a large number of whole-cell biocatalysts have been discovered and developed for HMF conversion into BHMF, FDCA and HMFCA, however there are no reports on microbial production of DFF and FFCA. While the production of BHMF and HMFCA mainly relies on wild type microorganisms, FDCA production, which requires multiple bioconversion steps from HMF, is strongly dependent on genetic engineering strategies. Together, the information gathered supports the possibility for the development of cell factories to produce high-value compounds, envisioning economical viable biorefineries., This research was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020, and by the Ministry of Science and Innovation (MICIN) through the grant RYC2020-030690-I (to AR)., info:eu-repo/semantics/publishedVersion

The establishment of lignocellulosic biorefineries is dependent on microorganisms being able to cope with the stressful conditions resulting from the release of inhibitory compounds during biomass processing. The yeast Kluyveromyces marxianus has been explored as an alternative microbial factory due to its thermotolerance and ability to natively metabolize xylose. The lignocellulose-derived inhibitors furfural and 5-hydroxymethylfurfural (HMF) are considered promising building-block platforms that can be converted into a wide variety of high-value derivatives. Here, several K. marxianus strains, isolated from cocoa fermentation, were evaluated for xylose consumption and tolerance towards acetic acid, furfural, and HMF. The potential of this yeast to reduce furfural and HMF at high inhibitory loads was disclosed and characterized. Our results associated HMF reduction with NADPH while furfural-reducing activity was higher with NADH. In addition, furans’ inhibitory effect was higher when combined with xylose consumption. The furan derivatives produced by K. marxianus in different conditions were identified. Furthermore, one selected isolate was efficiently used as a whole-cell biocatalyst to convert furfural and HMF into their derivatives, furfuryl alcohol and 2,5-bis(hydroxymethyl)furan (BHMF), with high yields and productivities. These results validate K. marxianus as a promising microbial platform in lignocellulosic biorefineries.

The aim of this work was to assess the occupational dose received by an interventional radiologist (IR) during computed tomography fluoroscopy (CTF)-guided procedures; to identify the most exposed areas of the body including the hands and fingers; to suggest recommendations for individual monitoring; and to improve radiation safety of the practice. A total of 53 CTF-guided procedures were studied. Twelve whole-body dosimeters were worn by the IR in each procedure for the assessment of the personal dose equivalent, Hp(10), on the chest, waist, and back, both over and under the lead apron, as well as the personal dose equivalent, Hp(0.07), on both arms, knees, and feet. Special gloves with casings to fit extremity dosimeters were prepared to assess Hp(0.07) to the fingers. The measured chest dose values were higher than those on the waist and back; the dominant hand or the left side was the most exposed. In general, the ring, middle, and index fingers of the dominant hand were the most exposed (maximum in the 36–39 mSv range), while wrist dose was negligible compared to finger doses. Based on the results obtained the following recommendations are suggested: protective devices (lead aprons, thyroid shield, and goggles) should be worn; Hp(10) should be assessed at the chest level both above and below the lead apron; finger doses can be measured on the basis of each middle finger; the arm closer to the beam should be monitored; and finally, a wrist dosimeter will not provide useful information.

Fragmentation of transient negative ions of tryptophan molecules formed through electron transfer in collisions with potassium atoms is presented for the first time in the laboratory collision energy range of 20 up to 100 eV. In the unimolecular decomposition process, the dominating side-chain fragmentation channel is assigned to the dehydrogenated indoline anion, in contrast to dissociative electron attachment of free low-energy electrons to tryptophan. The role of the collision complex formed by the potassium cation and tryptophan negative ion in the electron transfer process is significant for the mechanisms that operate at lower collision energies. At those collision times, on the order of a few tens of fs, the collision complex may not only influence the lifetime of the anion but also stabilize specific transition states and thus alter the fragmentation patterns considerably. DFT calculations, at the BHandHLYP/6-311++G(3df,2pd) level of theory, are used to explore potential reaction pathways and the evolvement of the charge distribution along those., F.F.d.S., T.C., and A.R. acknowledge the Portuguese National Funding Agency FCT-MCTES for IF-FCT IF/00380/2014, SFRH/BD/52538/2014, and PD/BD/114449/2016 and together with P.L.-V. the research grants PTDC/FIS-AQM/31215/2017 and PTDC/FIS-AQM/31281/2017. This work was also supported by Radiation Biology and Biophysics Doctoral Training Programme (RaBBiT, PD/00193/2012); UIDB/00068/2020 (CEFITEC) and UIDB/04378/2020 (UCIBIO). M.J.C. and A.G. also thank FCT-MCTES UIDB/04046/2020 and UIDP/04046/2020, and A.G. thanks the SFRH/BPD/89722/2012 grant. G.G. is partially funded by the Spanish Ministerio de Ciencia, Innovacion y Universidades (project no. PID2019-104727RB-C21) and CSIC (Project LINKA20085). O.I. acknowledges the Icelandic Center of Research (RANNIS) and the University of Iceland Research Fund for financial support. The authors thank Ragnar Bjornsson for fruitful discussions while preparing this manuscript., Pre-print

Seabirds are extensively used as environmental biomonitors and feathers are among the most analyzed matrices because they are one of the main excretory pathways to detoxify the bird's body of environmental contaminants. Still, there is a variation in contamination level between the different feathers of seabird species, driven by diet and physiology, such as molt strategy and feather formation sequence. We measured total mercury (THg) concentration in different types of feathers (wing, tail, ventral and dorsal) of the same individual in adults and juveniles of brown boobies (Sula leucogaster) from the northern coast of Rio de Janeiro state, Brazil. Brown booby had higher mean THg concentration (μg.g

4506 Background: Neoadjuvant chemotherapy is the standard of care in muscle-invasive bladder cancer patients. However, treatment is intense, the overall benefit is small, and there is no established marker to identify patients who benefit most. The aim of the study is to characterize cell-free DNA (cfDNA) methylation from patients receiving neoadjuvant chemotherapy in SWOG S1314, a prospective cooperative group trial, and to correlate the methylation signatures with pathologic response. Methods: Blood samples were collected prospectively from 73 patients before and during standard neoadjuvant chemotherapy. At radical cystectomy, pathologic response was documented. Plasma cfDNA was profiled using Infinium MethylationEPIC BeadChip array. Differential methylation between pathologic responders (≤pT1N0M0) and non-responders was analyzed, and a Random Forest model was used to generate a classifier predictive of treatment response. Results: Using pre-chemotherapy plasma cfDNA, we developed a methylation-based response score (mR-score) predictive of pathologic response. The mR-score also could be calculated using plasma samples collected after the first cycle of neoadjuvant chemotherapy, resulting in a similar predictive ability. Furthermore, we used cfDNA methylation data to calculate the circulating bladder DNA fraction, which had a modest but independent predictive ability for treatment response. When we combined the mR-score and circulating bladder DNA fraction, we successfully predicted pathologic response outcomes in 79% of patients based on their plasma collected before chemotherapy and after 1 cycle of chemotherapy. Conclusions: Our study provides proof of concept that cfDNA methylation may be used to predict treatment response in bladder cancer patients receiving neoadjuvant chemotherapy. Clinical trial information: NCT02177695.

Background Breast cancer is the main cause of female death by neoplasia in Brazil. Although half of the Brazilian population is black/brown (BB), socio-economic disparities translate in a vulnerable situation to those women. Access to health care is an important barrier to improve the health of BB women. This study aims to investigate trends in breast cancer mortality rates regarding race and age. Methods This is a population-based study of trends evaluation on breast cancer mortality in São Paulo state, Brazil, from 2000 to 2017. The absolute number of deaths and population figures (including race) by age-groups and years were available online from government data. Data on race were not available by ten-year age-groups, so the figures were projected according to the female age structure by year. Total rates by year and race were age-adjusted to the 'World Population (2000)'. For trend analysis, linear regression was used, with 5% level of significance. Results In the period were observed 60,940 breast cancer deaths, 76.7% in white and 17.5% in BB women. The absolute number of deaths in white and BB women was respectively 2,095 and 333 in 2000, and 3,076 and 999 in 2017. The total age-adjusted mortality rates per 100,000 women of white and BB in 2000 were respectively 16.4, 17.2 and 7.5. In 2017 rates were 14.6, 16.6 and 16.1. There was a trend towards reducing the mortality rates of white women (P = 0.002) and in their age-groups from 40 to 79 years (P Conclusions Although breast cancer figures of death and mortality rates in BB women have more than doubled in 18 years, rates reached almost the same figures of white women in the period. Changes in behaviour risk factors may explain this result. However, it is very likely that access to health care to these women has been improved, reducing the disparities in the health system. Key messages Breast cancer mortality rates in black/brown women have reached almost the same figures as white women from 2000 to 2017 in São Paulo, Brazil. Access to health care in black women may have improved in São Paulo, Brazil.

After nearly a century of vaccination and six decades of drug therapy, tuberculosis (TB) kills more people annually than any other infectious disease. Substantial challenges to disease eradication remain among vulnerable and underserved populations. The Guarani-Kaiowá people are an indigenous population in Paraguay and the Brazilian state of Mato Grosso do Sul. This community, marginalized in Brazilian society, experiences severe poverty. Like other South American indigenous populations, their TB prevalence is high, but the disease has remained largely unstudied in their communities. Herein, Mycobacterium tuberculosis isolates from local clinics were whole genome sequenced, and a population genetic framework was generated. Phylogenetics show M. tuberculosis isolates in the Guarani-Kaiowá people cluster away from selected reference strains, suggesting divergence. Most cluster in a single group, further characterized as M. tuberculosis sublineage 4.3.3. Closer analysis of SNPs showed numerous variants across the genome, including in drug resistance-associated genes, and with many unique changes fixed in each group. We report that local M. tuberculosis strains have acquired unique polymorphisms in the Guarani-Kaiowá people, and drug resistance characterization is urgently needed to inform public health to ensure proper care and avoid further evolution and spread of drug-resistant TB.

In‐phantom and in‐vivo three dimensional conformal radiation therapy (3DCRT) and volumetric modulated arc therapy (VMAT) skin doses, measured with and without bolus in a female anthropomorphic phantom RANDO and in patients, were compared against treatment planning system calculated values. A thorough characterization of the metal oxide semiconductor field effect transistor measurement system was performed prior to the measurements in phantoms and patients. Patients with clinical indication for postoperative external radiotherapy were selected. Skin dose showed higher values with 3DCRT technique compared with VMAT. The increase in skin dose due to the use of bolus was quantified. It was observed that, in the case of VMAT, the bolus effect on the skin dose was considerable when compared with 3DCRT. From the point of view of treatment time, bolus cost, and positioning reproducibility, the use of bolus in these situations can be optimized.

Negative ion formation in electron transfer experiments from fast neutral potassium (K) atom collisions with neutral tetrachloromethane (CCl

Purpose The purpose of this study was to evaluate the efficacy of needle holders in reducing staff hand exposure during biopsies guided by computed tomography fluoroscopy (CTF), through the analysis of data acquired during a detailed monitoring study, undertaken in parallel with an ongoing optimization process to reduce hand irradiation. Methods Hand monitoring was performed with 11 extremity detectors, two per finger (base and tip) and one on the back of the wrist, for the left (dominant) hand, during two series of biopsies with comparable characteristics. The first series (47 biopsies) were performed with only quick‐check method (QC) and occasional side‐handle (SH) manipulation of the needle. The second series (63 biopsies) were performed after introducing needle holders (NH) in the course of an optimization process. Results Choice of technique (QC, QC + NH, QC + SH) by the interventional radiologist (IR) was related to biopsy difficulty. Measured hand exposure was low (< 1 mSv) for all QC‐only procedures, and for most of the QC + NH procedures. Occasional side‐handle manipulation still occurred during challenging biopsies, so that 8% of biopsies in the second series accounted for ~70% of total fingertip dose (~90 mSv). The methodology used allowed a detailed insight into the dose reduction achievable with needle holders during real procedures, without the limitations of phantom measurements. Conclusions Needle holders proved effective in reducing mean hand exposure during clinical procedures where real‐time manipulation was necessary. Occasional side‐handle manipulation was found to contribute disproportionately to hand exposure. This highlights the importance of individual hand monitoring during CTF guided procedures.

Summary Background The sepsis burden on acute care services in middle-income countries is a cause for concern. We estimated incidence, prevalence, and mortality of sepsis in adult Brazilian intensive care units (ICUs) and association of ICU organisational factors with outcome. Methods We did a 1-day point prevalence study with follow-up of patients in ICU with sepsis in a nationally representative pseudo-random sample. We produced a sampling frame initially stratified by geographical region. Each stratum was then stratified by hospitals' main source of income (serving general public vs privately insured individuals) and ICU size (ten or fewer beds vs more than ten beds), finally generating 40 strata. In each stratum we selected a random sample of ICUs so as to enrol the total required beds in 1690 Brazilian adult ICUs. We followed up patients until hospital discharge censored at 60 days, estimated incidence from prevalence and length of stay, and generated national estimates. We assessed mortality prognostic factors using random-effects logistic regression models. Findings On Feb 27, 2014, 227 (72%) of 317 ICUs that were randomly selected provided data on 2632 patients, of whom 794 had sepsis (30·2 septic patients per 100 ICU beds, 95% CI 28·4–31·9). The ICU sepsis incidence was 36·3 per 1000 patient-days (95% CI 29·8–44·0) and mortality was observed in 439 (55·7%) of 788 patients (95% CI 52·2–59·2). Low availability of resources (odds ratio [OR] 1·67, 95% CI 1·02–2·75, p=0·045) and adequacy of treatment (OR 0·56, 0·37–0·84, p=0·006) were independently associated with mortality. The projected incidence rate is 290 per 100 000 population (95% CI 237·9–351·2) of adult cases of ICU-treated sepsis per year, which yields about 420 000 cases annually, of whom 230 000 die in hospital. Interpretation The incidence, prevalence, and mortality of ICU-treated sepsis is high in Brazil. Outcome varies considerably, and is associated with access to adequate resources and treatment. Our results show the burden of sepsis in resource-limited settings, highlighting the need to establish programmes aiming for sepsis prevention, early diagnosis, and adequate treatment. Funding Fundacao de Apoio a Pesquisa do Estado de Sao Paulo (FAPESP).