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Abstract PROTAC® (proteolysis-targeting chimera) molecules induce proximity between an E3 ligase and protein-of-interest (POI) to target the POI for ubiquitin-mediated degradation. Cooperative E3-PROTAC-POI complexes have potential to achieve neo-substrate selectivity beyond that established by POI binding to the ligand alone. Here, we extend the collection of ubiquitin ligases employable for cooperative ternary complex formation to include the C-degron E3 KLHDC2. Ligands were identified that engage the C-degron binding site in KLHDC2, subjected to structure-based improvement, and linked to JQ1 for BET-family neo-substrate recruitment. Consideration of the exit vector emanating from the ligand engaged in KLHDC2’s U-shaped degron-binding pocket enabled generation of SJ46421, which drives formation of a remarkably cooperative, paralog-selective ternary complex with BRD3BD2. Meanwhile, screening pro-drug variants enabled surmounting cell permeability limitations imposed by acidic moieties resembling the KLHDC2-binding C-degron. Selectivity for BRD3 compared to other BET-family members is further manifested in ubiquitylation in vitro, and prodrug version SJ46420-mediated degradation in cells. Selectivity is also achieved for the ubiquitin ligase, overcoming E3 auto-inhibition to engage KLHDC2, but not the related KLHDC1, KLHDC3, or KLHDC10 E3s. In sum, our study establishes neo-substrate-specific targeted protein degradation via KLHDC2, and provides a framework for developing selective PROTAC protein degraders employing C-degron E3 ligases.

With increased worldwide energy demand and carbon dioxide emissions from the use of fossil fuels, severe problems are being experienced in modern times. Energy is one of the most important resources for humankind, and its needs have been drastically increasing due to energy consumption, the rapid depletion of fossil fuels, and environmental crises. Therefore, it is important to identify and search for an alternative to fossil fuels that provides energy in a reliable, constant, and sustainable way that could use available energy sources efficiently for alternative renewable sources of fuel that are clean, non-toxic, and eco-friendly. In this way, there is a dire need to develop technologies for biofuel production with a focus on economic feasibility, sustainability, and renewability. Several technologies, such as biological and thermochemical approaches, are derived from abundant renewable biological sources, such as biomass and agricultural waste, using advanced conversion technologies for biofuel production. Biofuels are non-toxic, biodegradable, and recognized as an important sustainable greener energy source to conventional fossil fuels with lower carbon emissions, combat air pollution, empower rural communities, and increase economic growth and energy supply. The purpose of this review is to explain the basic aspects of biofuels and their sustainability criteria, with a particular focus on conversion technologies for biofuel production, challenges, and future perspectives.

Abstract Nuclear receptors are ligand-activated transcription factors that can often be useful drug targets. Unfortunately, ligand promiscuity leads to two-thirds of receptors remaining clinically untargeted. PXR is a nuclear receptor that can be activated by diverse compounds to elevate metabolism, negatively impacting drug efficacy and safety. This presents a barrier to drug development because compounds designed to target other proteins must avoid PXR activation while retaining potency for the desired target. This problem could be avoided by using PXR antagonists, but these compounds are rare, and their molecular mechanisms remain unknown. Here, we report structurally related PXR-selective agonists and antagonists and their corresponding co-crystal structures to describe mechanisms of antagonism and selectivity. Structural and computational approaches show that antagonists induce PXR conformational changes incompatible with transcriptional coactivator recruitment. These results guide the design of compounds with predictable agonist/antagonist activities and bolster efforts to generate antagonists to prevent PXR activation interfering with other drugs.

Introduction: The diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma (NHL) and, despite all the progress in this field, central nervous system infiltration (CNSi) still occurs at an incidence of 2–10%. The objective of the present study was to evaluate the Central Nervous System International Prognostic Index (CNS-IPI) score in daily practice regarding the reproducibility in a heterogeneous cohort apart from a clinical trial. Methods: Primary DLBCL patients were eligible for this study, between January 2007 and January 2017. All patients were treated with rituximab-based chemotherapy, mostly R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone). The CNSi was diagnosed by liquor (positive cytology and/or immunophenotype), computerized tomography, magnetic resonance image and/or fluorodeoxy-glucose-positron emission tomography, requested only in symptomatic patients when the CNSi was clinically suspected. The CNS-IPI was assessed by graphical comparison and calibration. Results: After applying the inclusion/exclusion criteria, 322 patients were available for the analysis. The median follow-up was 60 months and the median age was 58 years. Seven patients experienced CNSi, characterizing an incidence of 2.17% (7/322). Comparing groups of patients with and without CNSi, we observed that the lactate dehydrogenase (LDH), number of extranodal sites, IPI, kidney/adrenal and absence of complete response were statistically different. The CNS-IPI model stratified patients in a three-risk group model as low-, intermediate- and high-risk. In our cohort, using the same stratification, we obtained an equivalent the 2-year rate of CNS relapse of 0.0%, 0.8% and 13.8%, respectively. Conclusion: Our study reinforces the reproducibility of the CNS-IPI, specifically apart from clinical trials, and suggests the CNS-IPI score as a tool to guide therapy.

ABSTRACT Copious amounts of methane, a major constituent of greenhouse gases currently driving climate change, are emitted by livestock, and efficient methods that curb such emissions are urgently needed to reduce global warming. When fed to cows, the red seaweed Asparagopsis taxiformis (AT) can reduce enteric methane emissions by up to 80%, but the achieved results can vary widely. Livestock produce methane as a byproduct of methanogenesis, which occurs during the breakdown of feed by microbes in the rumen. The ruminant microbiome is a diverse ecosystem comprising bacteria, protozoa, fungi, and archaea, and methanogenic archaea work synergistically with bacteria to produce methane. Here, we find that an effective reduction in methane emission by high-dose AT (0.5% dry matter intake) was associated with a reduction in methanol-utilizing Methanosphaera within the rumen, suggesting that they may play a greater role in methane formation than previously thought. However, a later spike in Methanosphaera suggested an acquired resistance, possibly via the reductive dehalogenation of bromoform. While we found that AT inhibition of methanogenesis indirectly impacted ruminal bacteria and fermentation pathways due to an increase in spared H2, we also found that an increase in butyrate synthesis was due to a direct effect of AT on butyrate-producing bacteria such as Butyrivibrio, Moryella, and Eubacterium. Together, our findings provide several novel insights into the impact of AT on both methane emissions and the microbiome, thereby elucidating additional pathways that may need to be targeted to maintain its inhibitory effects while preserving microbiome health and animal productivity.IMPORTANCELivestock emits copious quantities of methane, a major constituent of the greenhouse gases currently driving climate change. Methanogens within the bovine rumen produce methane during the breakdown of feed. While the red seaweed Asparagopsis taxiformis (AT) can significantly reduce methane emissions when fed to cows, its effects appear short-lived. This study revealed that the effective reduction of methane emissions by AT was accompanied by the near-total elimination of methane-generating Methanosphaera. However, Methanosphaera populations subsequently rebounded due to their ability to inactivate bromoform, a major inhibitor of methane formation found in AT. This study presents novel findings on the contribution of Methanosphaera to ruminal methanogenesis, the mode of action of AT, and the possibility for complementing different strategies to effectively curb methane emissions.

Abstract Robotic milking systems are successful innovations in the development of dairy cattle. The objective of this study was to analyse the milking characteristics and behavior of dairy cows of different calving orders in “milk first” robotic milking systems. The data were collected from a commercial herd located in the Midwest region of Minas Gerais (Brazil), which uses an automatic milking system (AMS TM, DeLaval). Were analysed 26,574 observations of 235 Holstein cows were available. Data were evaluated by multivariate analysis of variance and the Tukey test. - Tthe characteristics milk flow and milking efficiency were more favourable for multiparous cows (p

To assess the clinical impact of delayed testosterone recovery (TR) following the discontinuation of medical androgen deprivation therapy (ADT), a retrospective, longitudinal analysis was conducted in adult males with prostate cancer using the Optum® de-identified Electronic Health Record data set and Optum® Enriched Oncology Data (2010–2021). Of 3875 patients who initiated and discontinued ADT, 1553 received one or more testosterone-level tests within the 12 mo following discontinuation and were included in this study. These 1553 patients were categorized into two cohorts: 25% as TR (testosterone levels >280 ng/dl at any test within 12 mo following ADT discontinuation) and 75% as non-TR. At baseline, non-TR patients were older, had lower testosterone levels, and were more likely to have diabetes, hyperlipidemia, and hypertension, but less likely to have sexual dysfunction. After adjustment for baseline characteristics, the TR cohort had a lower risk of new-onset diabetes (hazard ratio [HR] 0.47; 95% confidence interval [CI] 0.27–0.79), trended toward a lower risk of new-onset depression (HR 0.58; 95% CI 0.33–1.02), and had a higher likelihood of seeking treatment for sexual dysfunction (HR 1.33; 95% CI 0.99–1.78) versus the non-TR cohort. These findings support monitoring testosterone levels after ADT discontinuation to manage potential long-term comorbidities in patients with prostate cancer. Patient summary: This real-world analysis of males with prostate cancer who were treated with medical androgen deprivation therapy (ADT) found that most patients did not have their testosterone level checked in the 12 mo after stopping ADT. Of those who did, 75% did not achieve normal testosterone levels (>280 ng/dl), and these patients were more likely to experience new-onset diabetes than those who achieved normal testosterone levels. These results suggest that to ensure effective clinical decision-making, physicians should check patients’ testosterone levels after stopping ADT.

Bacillus Calmette-Guérin (BCG) is the first line treatment for bladder cancer and it is also proposed for melanoma immunotherapy. BCG modulates the tumor microenvironment (TME) inducing an antitumor effective response, but the immune mechanisms involved still poorly understood. The immune profile of B16-F10 murine melanoma cells was assessed by infecting these cells with BCG or stimulating them with agonists for different innate immune pathways such as TLRs, inflammasome, cGAS-STING and type I IFN. B16-F10 did not respond to any of those stimuli, except for type I IFN agonists, contrasting with bone marrow-derived macrophages (BMDMs) that showed high production of proinflammatory cytokines. Additionally, we confirmed that BCG is able to infect B16-F10, which in turn can activate macrophages and spleen cells from mice in co-culture experiments. Furthermore, we established a subcutaneous B16-F10 melanoma model for intratumoral BCG treatment and compared wild type mice to TLR2-/-, TLR3-/-, TLR4-/-, TLR7-/-, TLR3/7/9-/-, caspase 1-/-, caspase 11-/-, IL-1R-/-, cGAS-/-, STING-/-, IFNAR-/-, MyD88-/-deficient animals. These results in vivo demonstrate that MyD88 signaling is important for BCG immunotherapy to control melanoma in mice. Also, BCG fails to induce cytokine production in the co-culture experiments using B16-F10 and BMDMs or spleen cells derived from MyD88-/- compared to wild-type (WT) animals. Immunotherapy with BCG was not able to induce the recruitment of inflammatory cells in the TME from MyD88-/- mice, impairing tumor control and IFN-γ production by T cells. In conclusion, MyD88 impacts on both innate and adaptive responses to BCG leading to an efficient antitumor response against melanoma.

IntroductionGuanylate-binding proteins (GBPs) are produced in response to pro-inflammatory signals, mainly interferons. The most studied cluster of GBPs in mice is on chromosome 3. It comprises the genes for GBP1-to-3, GBP5 and GBP7. In humans, all GBPs are present in a single cluster on chromosome 1. Brucella abortus is a Gram-negative bacterium known to cause brucellosis, a debilitating disease that affects both humans and animals. Our group demonstrated previously that GBPs present on murine chromosome 3 (GBPchr3) is important to disrupt Brucella-containing vacuole and GBP5 itself is important to Brucella intracellular LPS recognition. In this work, we investigated further the role of GBPs during B. abortus infection.Methods and resultsWe observed that all GBPs from murine chromosome 3 are significantly upregulated in response to B. abortus infection in mouse bone marrow-derived macrophages. Of note, GBP5 presents the highest expression level in all time points evaluated. However, only GBPchr3-/- cells presented increased bacterial burden compared to wild-type macrophages. Brucella DNA is an important Pathogen-Associated Molecular Pattern that could be available for inflammasome activation after BCV disruption mediated by GBPs. In this regard, we observed reduced IL-1β production in the absence of GBP2 or GBP5, as well as in GBPchr3-/- murine macrophages. Similar result was showed by THP-1 macrophages with downregulation of GBP2 and GBP5 mediated by siRNA. Furthermore, significant reduction on caspase-1 p20 levels, LDH release and Gasdermin-D conversion into its mature form (p30 N-terminal subunit) was observed only in GBPchr3-/- macrophages. In an in vivo perspective, we found that GBPchr3-/- mice had increased B. abortus burden and higher number of granulomas per area of liver tissue, indicating increased disease severity.Discussion/conclusionAltogether, these results demonstrate that although GBP5 presents a high expression pattern and is involved in inflammasome activation by bacterial DNA in macrophages, the cooperation of multiple GBPs from murine chromosome 3 is necessary for full control of Brucella abortus infection.

Napier grass (Pennisetum purpureum Schumach) comprises up to 80% of the cattle diet in many tropical and subtropical regions and is used primarily by smallholder farmers. Despite the grass’s high yield, resulting animal productivity from this grass is low. One of the key reasons for the low animal productivity of Napier grass is its low nutritive value under current management. Taken together, previous work has shown the current yield, crude protein (CP), and metabolisable energy (ME) of Napier grass to be 26 t dry matter (DM)/ha/year, 96 g/kg DM, and 8.7 MJ/kg DM, respectively, ranging from 2 to 86 t DM/ha/year, 9 to 257 g CP/kg DM, and 5.9 to 10.8 MJ ME/kg DM, respectively, suggesting an opportunity for significant improvement on both yield and nutritive value of this grass. The DM yield and nutritive value of this grass are inversely related, indicating a trade-off between yield and quality; however, this trade-off could be minimised by increasing sowing density and harvesting frequency. Available literature shows that this simple management strategy of increasing sowing density (50 cm × 40 cm) and harvesting frequency (11–12 harvests/year) provides 71 t DM/ha with 135 g/kg DM CP and 10.8 MJ ME/kg DM. This quality of Napier grass has the potential to increase both milk and meat production substantially in the tropics and subtropics, and the farmers will likely find this simple management acceptable due to the high yield obtained through this management. However, there is a paucity of work in this field. Therefore, management strategies to improve the nutritive value of Napier grass are required to increase milk and meat production in the tropics and subtropics and in doing so improve the food security of more than half of the global population living in these regions.

Here, de Sá et al. show that Gasdermin-D is transiently activated in Leishmania-infected macrophages and promotes NLRP3 inflammasome activation, but not cell death. Gasdermin-D is cleaved into a noncanonical fragment, indicating that Leishmania subverts Gasdermin-D-mediated host response to establish leishmaniasis.

Napier grass (Pennisetum purpureum Schumach) supports a significant proportion of animal production in subtropical and tropical regions, but its quality is low and when offered alone, results in low ruminant production. Shifting the management of Napier grass towards a higher-quality feed increased milk yield and liveweight gain for small, mature cattle without supplementation. This review highlights the opportunity for further increases in milk and meat production for differing classes of livestock in the tropics and subtropics by improving the nutritive value of Napier grass using new best management practice flowing on to improve food security for the millions of people in these regions.

As a result of the depletion of fossil resources, ongoing population growth, and the industrialized economy, energy demand has been rising quickly throughout the world. India is now the world’s third-largest oil consumer, surpassing Japan and Russia. Today, biofuel research is conducted worldwide because surrounding two essential characteristics: sustainability and renewability. Biofuels have gained considerable significance as a result of dwindling oil sources, worries about energy security, and the escalating environmental issues associated with climate change and greenhouse gas emissions. In most cases, biofuels are produced by subjecting materials that have been densified to the process of heat conversion. In the disciplines of research and development, alternative energy development is a top focus. Due to the depletion of fossil fuel resources, it has become important to find innovative replacements for fossil fuels, such as biofuels, to generate heat and power. Biofuels may be generated using several methodologies, encompassing biological, chemical, and physical approaches. The three steps of densification systems’ pre-, during-, and post-pelletization procedures convert biomass into pellets. Several agricultural wastes, such as grain dust, crop leftovers, and fruit tree residues, are available as sources of agricultural energy. Bioenergy from biomass, such as leftovers and energy crops, can be used to produce contemporary energy carriers. This article focuses on an overview of sustainable and renewable biofuel resources and their commercialization.

Despite the importance of the respiratory route for Brucella transmission, the lung immune response to this pathogen is scarcely characterized. We investigated the role of the cGAS/STING pathway of microbial DNA recognition in the control of respiratory Brucella infection. After in vitro B. abortus infection, CFU numbers were significantly higher in alveolar macrophages (AM) and lung explants from STING KO mice than in samples from wild type (WT) mice, but no difference was observed for cGAS KO samples. CFU were also increased in WT AM and lung epithelial cells preincubated with the STING inhibitor H151. Several proinflammatory cytokines (TNF-α, IL-1β, IL-6, IP-10/CXCL10) were diminished in Brucella-infected lung explants and/or AM from STING KO mice and cGAS KO mice. These cytokines were also reduced in infected AM and lung epithelial cells pretreated with H151. After intratracheal infection with B. abortus, STING KO mice exhibited increased CFU in lungs, spleen and liver, a reduced expression of IFN-β mRNA in lungs and spleen, and reduced levels of proinflammatory cytokines and chemokines in bronchoalveolar lavage fluid (BALF) and lung homogenates. Increased lung CFU and reduced BALF cytokines were also observed in cGAS KO mice. In summary, the cGAS/STING pathway induces the production of proinflammatory cytokines after respiratory Brucella infection, which may contribute to the STING-dependent control of airborne brucellosis.

Antecedentes/Objetivo: La dificultad para mantener una rutina constante de ejercicio ha sido ampliamente descrita en pacientes con diabetes. Esto ha llevado a la búsqueda de estrategias para integrar la actividad física y el ejercicio como parte del tratamiento. Método: Se aplicaron el Cuestionario Global de Actividad Física (GPAQ) y un cuestionario para determinar las barreras que les impedían hacer ejercicio al inicio y a los 2, 3 y 12 meses. Cada evaluación estableció estrategias específicas para resolver la barrera para hacer ejercicio. Resultados: Se incluyeron 154 pacientes con diabetes tipo 2, el 55.6% mujeres, con una edad de 52,6 ± 10 años y un tiempo de diagnóstico de la diabetes de 1.5 (0-3) años. Al inicio, el 74.5% de los pacientes tenían una o más barreras para hacer ejercicio. Las barreras disminuyeron después de la intervención al 39.3%. La barrera más frecuente fue «falta de tiempo» (54%). Al año, el 48.4% de los pacientes no tenían barreras, y los pacientes sin barreras aumentaron el tiempo de ejercicio moderado de 45 (0-155) a 160 (80-240) minutos al año. Conclusiones: Los programas deben individualizarse según la principal barrera, pero sin descuidar otras barreras que puedan surgir en función de la evaluación del paciente.

Purpose: The aortic time-enhancement curve obtained from dynamic CT myocardial perfusion imaging can be used to derive the cardiac output (CO) index based on the indicator dilution principle. The objective of this study was to investigate the effect of cardiac phase at which CT myocardial perfusion imaging is triggered on the CO index measurement with this approach. Methods: Electrocardiogram (ECG) gated myocardial perfusion imaging was performed on farm pigs with consecutive cardiac axial scans using a large-coverage CT scanner (Revolution, GE Healthcare) after intravenous contrast administration. Multiple sets of dynamic contrast-enhanced (DCE) cardiac images were reconstructed retrospectively from 30% to 80% R-R intervals with a 5% phase increment. The time-enhancement curve sampled from above the aortic orifice in each DCE image set was fitted with a modified gamma variate function (MGVF). The fitted curve was then normalized to the baseline data point unaffected by the streak artifact emanating from the contrast solution in the right heart chamber. The Stewart–Hamilton equation was used to calculate the CO index based on the integral of the fitted normalized aortic curve, and the results were compared among different cardiac phases. Results: The aortic time-enhancement curves sampled at different cardiac phases were different from each other, especially in the baseline portion of the curve where the effect of streak artifact was prominent. After properly normalizing and denoising with a MGVF, the integrals of the aortic curve were minimally different among cardiac phases (0.228 ± 0.001 Hounsfield Unit × second). The corresponding mean CO index was 4.031 ± 0.028 L/min. There were no statistical differences in either the integral of the aortic curve or CO index among different cardiac phases (p > 0.05 for all phases).

The bacillus Calmette–Guérin (BCG) is an attenuated bacterium derived from virulent Mycobacterium bovis. It is the only licensed vaccine used for preventing severe forms of tuberculosis in children. Besides its specific effects against tuberculosis, BCG administration is also associated with beneficial non-specific effects (NSEs) following heterologous stimuli in humans and mice. The NSEs from BCG could be related to both adaptive and innate immune responses. The latter is also known as trained immunity (TI), a recently described biological feature of innate cells that enables functional improvement based on metabolic and epigenetic reprogramming. Currently, the mechanisms related to BCG-mediated TI are the focus of intense research, but many gaps are still in need of elucidation. This review discusses the present understanding of TI induced by BCG, exploring signaling pathways that are crucial to a trained phenotype in hematopoietic stem cells and monocytes/macrophages lineage. It focuses on BCG-mediated TI mechanisms, including the metabolic-epigenetic axis and the inflammasome pathway in these cells against intracellular pathogens. Moreover, this study explores the TI in different immune cell types, its ability to protect against various intracellular infections, and the integration of trained innate memory with adaptive memory to shape next-generation vaccines.

La creciente epidemia de obesidad ha sido uno de los retos más importantes de salud pública en México durante los últimos años. Con apoyo de la Federación Mundial de Obesidad, en 2021 formamos un grupo de profesionales para identificar y resumir las acciones prioritarias en las que puede enfocarse nuestro país para hacer frente a esta epidemia. Al proceso de desarrollo y discusión de este grupo se sumaron más de 1 000 profesionales de la salud para retomar recomendaciones de documentos y guías de alto nivel previamente publicados. En conmemoración del Día Mundial de la Obesidad, en este 2022 se presenta esta postura como insumo para el desarrollo de acciones en el ámbito profesional y de los diferentes sectores, en la que se incluyen 10 recomendaciones de acción, desde la perspectiva poblacional hasta la atención individualizada, y se enfatiza en la importancia de la participación social, de las intervenciones integrales con visión centrada en la persona y de la sostenibilidad planetaria, además de mejorar la educación y las campañas de difusión, propiciar un ambiente promotor de entornos activos y blindar de conflictos de interés los esfuerzos de prevención y control. La postura hace un llamado para abordar la obesidad de manera seria, con base en la evidencia científica, oportuna e integral, con enfoque de curso de vida, de forma ética y sensible, y sin perpetuar las barreras del estigma de peso en la sociedad.

Endoplasmic reticulum (ER) stress plays a major role in several inflammatory disorders. ER stress induces the unfolded protein response (UPR), a conserved response broadly associated with innate immunity and cell metabolic function in various scenarios. Brucella abortus, an intracellular pathogen, triggers the UPR via Stimulator of interferon genes (STING), an important regulator of macrophage metabolism during B. abortus infection. However, whether ER stress pathways underlie macrophage metabolic function during B. abortus infection remains to be elucidated. Here, we showed that the UPR sensor inositol-requiring enzyme 1α (IRE1α) is as an important component regulating macrophage immunometabolic function. In B. abortus infection, IRE1α supports the macrophage inflammatory profile, favoring M1-like macrophages. IRE1α drives the macrophage metabolic reprogramming in infected macrophages, contributing to the reduced oxidative phosphorylation and increased glycolysis. This metabolic reprogramming is probably associated with the IRE1α-dependent expression and stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), an important molecule involved in cell metabolism that sustains the inflammatory profile in B. abortus-infected macrophages. Accordingly, we demonstrated that IRE1α favors the generation of mitochondrial reactive oxygen species (mROS) which has been described as an HIF-1α stabilizing factor. Furthermore, in infected macrophages, IRE1α drives the production of nitric oxide and the release of IL-1β. Collectively, these data unravel a key mechanism linking the UPR and the immunometabolic regulation of macrophages in Brucella infection and highlight IRE1α as a central pathway regulating macrophage metabolic function during infectious diseases.

PURPOSEWe present a physician survey of the impact of 21-gene Breast Recurrence Score test results on treatment decisions in clinical practice in Latin America.METHODSThis prospective survey enrolled consecutive patients at 14 sites in Argentina, Colombia, Mexico, and Peru who had routine 21-gene testing. Physician surveys captured patient and tumor characteristics and treatment decisions before and after 21-gene test results. The survey spanned the period before and after Trial Assigning Individualized Options for Treatment (TAILORx) results reported (June 2018). Overall net percent change in adjuvant chemotherapy recommendations was estimated, and asymptotic 95% CIs with continuity correction were calculated. The proportion with a change between pretest treatment recommendation and actual treatment received was calculated overall and by Recurrence Score groups per TAILORx.RESULTSBetween March 2015 and December 2019, the survey was completed for 647 patients; 20% were node-positive. The mean patient age was 54 years (24-85 years); 55% were postmenopausal; 17%, 63%, and 20% had grade 1, 2, and 3 tumors, respectively; and 30% had tumors > 2 cm. Recurrence Score (RS) results were as follows: 20% RS 0-10, 56% RS 11-25, and 24% RS 26-100. Overall, chemotherapy recommendations fell by a relative proportion of 39% (95% CI, 33.4 to 44.3) after 21-gene testing (33% decrease in node-negative and 55% decrease in node-positive). Among node-negative patients, the relative decrease in chemotherapy recommendations was 28% (95% CI, 18.9 to 39.5) before TAILORx and 36% (95% CI, 28.4 to 43.7) after.CONCLUSIONTo our knowledge, this large survey of 21-gene test practice patterns was the first conducted in Latin America and showed the relevance of 21-gene testing in low- and medium-resource countries to minimize chemotherapy overuse and underuse in breast cancer. The results showed substantial reductions in chemotherapy use overall—especially after TAILORx reported—indicating the practice-changing potential of that study.

PURPOSEWe evaluated the impact of 21-gene test results on treatment decisions for patients with early-stage breast cancer treated under the public health care system in Brazil, Sistema Único de Saúde.METHODSEligible patients treated at Hospital Pérola Byington and Santa Casa de Misericórdia de São Paulo in Brazil were required to have the following characteristics: postsurgery with hormone receptor–positive, human epidermal growth factor 2–negative, node-negative and node-positive, and T1/T2 breast cancer and patients with these characteristics were candidates for adjuvant systemic therapy. Treatment recommendations, chemotherapy plus hormonal therapy (CT + HT) or HT alone, were captured before and after 21-gene test results.RESULTSFrom August 2018 to April 2019, 179 women were enrolled. The mean age was 58 years (29-86 years), 135 (76%) were postmenopausal, and 58 (32%) had node-positive breast cancer. Most patients (61%) had a tumor > 2 cm, including 7% with tumors > 4 cm. Using Recurrence Score (RS) result cut points on the basis of the TAILORx trial, 40 (22%) had RS 0-10, 91 (51%) had RS 11-25, and 48 (27%) had RS 26-100. Before 21-gene testing, 162 of 179 (91%) patients were recommended for CT. After testing, 117 of 179 patients (65%) had changes in CT recommendation: 112 (63%) who were initially recommended CT received HT alone and five (3%) who were initially recommended HT alone received CT + HT. After 21-gene testing, 99% of physicians reported strong confidence in their treatment recommendations.CONCLUSIONThe change in clinical practice at these public hospitals was greater than expected: 66% of initial treatment recommendations were changed to omit CT with 21-gene test results. Clinicopathologic features did not correlate well with 21-gene test results and did not adequately identify those most likely to benefit from CT.

Milk production by dairy cows is sensitive to increased levels of stress hormones such as glucocorticoids (GC) that also regulate the transcription of several genes required for milk synthesis. Whereas previous studies identified that an exogenous GC such as dexamethasone (DEX) transiently suppresses milk yield in several species without any pronounced effect on milk protein or fat percentage, the mechanism underlying this effect has not been established. In this study we sought to establish changes within the mammary glands of non-pregnant dairy cows in their second lactation (n = 3–4; 648–838 kg) following a single dose of exogenous DEX. Changes in the udder were monitored by serial biopsy of alternating quarters, concurrent with quarter-level monitoring of milk yield and composition. Dexamethasone increased serum glucose levels from 12–36 h (p

Programmed cell death (PCD) is an important mechanism of innate immunity against bacterial pathogens. The innate immune PCD pathway involves the molecules caspase-7 and caspase-8, among others. Brucella abortus is a gram-negative bacterium that causes a zoonotic disease termed brucellosis. The innate immune response against this pathogen involves activation of inflammasome components and induction of pyroptosis. However, no studies so far have revealed the role of caspase-7 or caspase-8 during this bacterial infection. Herein, we demonstrate that caspase-7 is dispensable for caspase-1 processing, IL-1β secretion and cell death in macrophages. Additionally, caspase-7 deficient animals control B. abortus infection as well as the wild type mice. Furthermore, we addressed the role of caspase-8 in inflammasome activation and pyroptosis during this bacterial infection. Macrophages deficient in caspase-8 secreted reduced amounts of IL-1β that parallels with diminished caspase-1 activity when compared to wild type cells. Additionally, caspase-8 KO macrophages showed reduced LDH release when compared to wild type, suggesting that caspase-8 may play an important role in pyroptosis in response to B. abortus. Finally, caspase-8 KO animals were more susceptible to Brucella infection when compared to wild type mice. Overall, this study contributes to a better understanding of the involvement of caspase-7 and caspase-8 in innate immunity against B. abortus infection.

Free carriers and electrical polarization coexist in ferroelectric metals. Here, the authors use a capacitive method to probe the electronic compressibility of free carriers in a tunable semimetal, extract the polarized contribution, and study the carrier dependence of the ferroelectric state.

This study aimed to evaluate the viability of a new procedure based on machine learning (ML), virtual reality (VR), and implicit measures to discriminate empathy. Specifically, eye-tracking and decision-making patterns were used to classify individuals according to their level in each of the empathy dimensions, while they were immersed in virtual environments that represented social workplace situations. The virtual environments were designed using an evidence-centered design approach. Interaction and gaze patterns were recorded for 82 participants, who were classified as having high or low empathy on each of the following empathy dimensions: perspective-taking, emotional understanding, empathetic stress, and empathetic joy. The dimensions were assessed using the Cognitive and Affective Empathy Test. An ML-based model that combined behavioral outputs and eye-gaze patterns was developed to predict the empathy dimension level of the participants (high or low). The analysis indicated that the different dimensions could be differentiated by eye-gaze patterns and behaviors during immersive VR. The eye-tracking measures contributed more significantly to this differentiation than did the behavioral metrics. In summary, this study illustrates the potential of a novel VR organizational environment coupled with ML to discriminate the empathy dimensions. However, the results should be interpreted with caution, as the small sample does not allow general conclusions to be drawn. Further studies with a larger sample are required to support the results obtained in this study.

This study demonstrates the feasibility of a mobile aerial drone particle monitoring system (DPMS) to measure and detect changes in harvest dust levels based on moderate adjustments to harvester settings. When compared to an earlier harvester, a new harvester operated at standard settings produced 35% fewer PM2.5s, 32% fewer PM10s, and 42% fewer TSPs. Increasing the ground speed had an adverse effect on dust mitigation, while reducing it by half only offered a slightly more favorable margin. The mutual effects of some meteorological factors were found to be slightly correlated with PM10 and TSP readings and caused significant variability in PM2.5 readings. The current findings show similar trends to PM reduction estimates of previous studies, with only a nominal difference of 10 to 15% points. Overall, the DPMS was found to perform well within an acceptable statistical confidence level. The use of DPMSs could reduce the logistical needs, complexity issues, and feedback times often experienced using the Federal Reference Method (FRM). Further investigation is needed to verify its robustness and to develop potential correlations with the FRM under different orchard location and management practices. At this stage, the current aerial DPMS should be considered a rapid screening tool not to replace the FRM, but rather to complement it in evaluating the feasibility of dust abatement strategies for the almond industry.

The resistance of solid-separated solid wastes (SSDWs) to moderate airflow ranging from 0.05 to 0.30 m3/s-m2 was measured at various bed depths and moisture levels. The pressure drop across a loose-fill fixed bed column was observed to increase more rapidly with increasing airflow rates than with increasing bed depths. An increase in the moisture content (10 percentage points) caused a decrease in the pressure drop by an average of 13.2–17.0%, evaluated within a 10–40% moisture content (MC) range. A full-factorial model analysis using standard least squares was used to describe the main effects and interactions of the test parameters in predicting the pressure drop. The Hukill and Ives nonlinear model was able to accurately describe the airflow resistance data of SSDWs at various MCs. Empirical curves describing the SSDW resistance to airflow were developed to aid in the preliminary design of ventilation systems for drying and storage.

Abstract Bacillus Calmette-Guerin (BCG) is the only FDA approved first line therapy for patients with nonmuscle invasive bladder cancer. The purpose of this study is to better understand the role of innate immune pathways involved in BCG immunotherapy against murine bladder tumor. We first characterized the immunological profile induced by the MB49 mouse urothelial carcinoma cell line. MB49 cells were not able to activate an inflammatory response (TNF-α, IL-6, CXCL-10 or IFN-β) after the stimulus with different agonists or BCG infection, unlike macrophages. Although MB49 cells are not able to induce an efficient immune response, BCG treatment could activate other cells in the tumor microenvironment (TME). We evaluated BCG intratumoral treatment in animals deficient for different innate immune molecules (STING−/−, cGAS−/−, TLR2−/−, TLR3−/−, TLR4−/−, TLR7−/−, TLR9−/−, TLR3/7/9−/−, MyD88−/−, IL-1R−/−, Caspase1/11−/−, Gasdermin-D−/− and IFNAR−/−) using the MB49 subcutaneous mouse model. Only MyD88−/− partially responded to BCG treatment compared to wild type (WT) mice, suggesting a role played by this adaptor molecule. Additionally, BCG intratumoral treatment regulates cellular infiltrate in TME with an increase of inflammatory macrophages, neutrophils and CD8+ T lymphocytes, suggesting an immune response activation that favors tumor remission in WT mice but not in MyD88−/−. The experiments using MB49 cells infected with BCG and co-cultured with macrophages also demonstrated that MyD88 is essential for an efficient immune response. Our data suggests that BCG immunotherapy depends partially on the MyD88-related innate immune pathway.

A bench-scale 3-L internally illuminated concentric-tube airlift photobioreactor with an aspect ratio (H/D) ratio of approximately 2.0 and riser-to-downcomer area (Ar/Ad) ratio of 0.42 was designed, fabricated, and tested for the cultivation of Spirulina platensis. Using the results of the bench-scale experiments, a preliminary techno-economic analysis of producing dried Spirulina biomass in a larger scale (in four 20-m3 photobioreactors) was conducted. For this analysis, the aspect ratio and Ar/Ad were maintained. The selling price of the Spirulina powder was based on literature. For the preliminary techno-economic evaluation, 300 days per year production and continuous operation was assumed. Results show that Spirulina powder production plant with a total photobioreactor capacity of 80-m3 revealed an attractive venture. The profit margin is positive, and the payback period is 6.34 years. A sensitivity analysis was also conducted. The effects of total photobioreactor capacity and operating conditions on net income, profit margin, and payback period were determined. The minimum selling price at various total photobioreactor capacities were also calculated.

Systematic measurement of pasture biomass (kg DM/ha) is crucial for optimising pasture utilisation and increasing dairy farm profitability. On-farm pasture monitoring can be conducted using various sensors, but calibrations are necessary to convert the measured variable into pasture biomass. In this study, we conducted three experiments in New South Wales (Australia) to evaluate the use of the rising plate meter (RPM), pasture reader (PR), unmanned aerial vehicles (UAV) and satellites as pasture monitoring tools. We tested various calibration methods that can improve the accuracy of the estimations and be implemented more easily on-farm. The results indicate that UAV and satellite-derived reflectance indices (e.g., Normalised Difference Vegetation Index) can be indirectly calibrated with height measurements obtained from an RPM or PR. Height measurements can be then converted into pasture biomass ideally by conducting site-specific sporadic calibrations cuts. For satellites, using the average of the entire paddock, root mean square error (RMSE) = 226 kg DM/ha for kikuyu (Pennisetum clandestinum Hochst. ex Chiov) and 347 kg DM/ha for ryegrass (Lolium multiflorum L.) is as effective as but easier than matching NDVI pixels with height measurement using a Global Navigation Satellite System (RMSE = 227 kg DM/ha for kikuyu and 406 kg DM/ha for ryegrass). For situations where no satellite images are available for the same date, the average of all images available within a range of up to four days from the day ground measurements were taken could be used (RMSE = 225 kg DM/ha for kikuyu and 402 kg DM/ha for ryegrass). These methodologies aim to develop more practical and easier-to-implement calibrations to improve the accuracy of the predictive models in commercial farms. However, more research is still needed to test these hypotheses under extended periods, locations, and pasture species.

We propose a new statistical analysis of the Acoustic Emissions (AE) produced in a series of triaxial deformation experiments leading to fractures and failure of two different rocks, namely, Darley Dale Sandstone (DDS) and AG Granite (AG). By means of q-statistical formalism, we are able to characterize the pre-failure processes in both types of rocks. In particular, we study AE inter-event time and AE inter-event distance distributions. Both of them can be reproduced with q-exponential curves, showing universal features that are observed here for the first time and could be important in order to understand more in detail the dynamics of rock fractures.

Breast cancer (BC) is the most common type of cancer among women in almost all countries worldwide and is one of the oncological pathologies for which is indicated fertility preservation, a type of procedure used to help keep a person’s ability to have children. Follicular fluid (FF) is a major component of oocyte microenvironment, which is involved in oocyte growth, follicular maturation, and in communication between germ and somatic cells; furthermore, it accumulates all metabolites during oocytes growth. To obtain information about changes on fertility due to cancer, we aimed at investigating potential biomarkers to discriminate between FF samples obtained from 16 BC patients and 10 healthy women undergoing in vitro fertilization treatments. An NMR-based metabolomics approach was performed to investigate the FF metabolic profiles; ELISA and western blotting assays were used to investigate protein markers of oxidative and inflammatory stress, which are processes closely related to cancer. Our results seem to suggest that FFs of BC women display some significant metabolic alterations in comparison to healthy controls, and these variations are also related with tumor staging.

Abstract Background Malaria is one of the main causes of death in Angola, particularly among children under 5 years of age. An essential means to improve the situation is with strong malaria case management; this includes diagnosing suspected patients with a confirmatory test, either with a rapid diagnostic test (RDT) or microscopy, prompt and correct treatment with artemisinin-based combination therapy (ACT), and proper case registration (track). In 2011, the United States President’s Malaria Initiative (PMI) launched a country-wide programme to improve malaria case management through the provision of regular training and supervision at different levels of health care provision. An evaluation of malaria testing, treatment and registration practices in eight provinces, and at health facilities of various capacities, across Angola was conducted to assess progress of the national programme implementation. Methods A retrospective assessment analysed data collected during supervision visits to health facilities conducted between 2012 and 2016 in 8 provinces in Angola. The supervision tool used data collected for malaria knowledge, testing, treatment and case registration practices among health workers as well as health facilities stock outs from different levels of health care delivery. Contingency tables with Pearson chi-squared (χ 2) tests were used to identify factors associated with “knowledge”, “test”, “treat” and “track.” Multivariable logistic regression models were used to assess factors associated with the defined outcomes. Results A total of 7156 supervisions were conducted between September 2012 and July 2016. The overall knowledge, testing, treatment and tracking practices among health care workers (HCWs) increased significantly from 2013 to 2016. Health care workers in 2016 were 3.3 times (95% CI: 2.7–3.9) as likely to have a higher knowledge about malaria case management as in 2013 (p

The aim of this pilot study is to evaluate if SARS-CoV-2 infection or vaccination against SARS-CoV-2 infection induce observable metabolic effects in follicular fluid of women who are following in vitro fertilization (IVF) treatments. The possible impact of coronavirus disease 2019 (COVID-19) on fertility and IVF outcome is considered. We have selected for this study: six women vaccinated against SARS-CoV-2 infection, five recovered COVID-19 patients, and we used nine healthy women as the control group. At the time of oocytes retrieval from participants in the study, follicular fluids were collected and metabolomic analysis was performed by 1H NMR spectroscopy in combination with multivariate analysis to interpret the spectral data. The search for antibody positivity in the follicular fluid aspirates was also carried out, together with the western blotting analysis of some inflammatory proteins, interleukin-6, tumor necrosis factor α (TNFα), and the free radical scavenger superoxide dismutase 2. Higher levels of Ala and Pro together with lower levels of lipids and trimethylamine N-oxide (TMAO) were found in follicular fluids (FFs) of vaccinated women while lower levels of many metabolites were detected in FFs of recovered COVID patients. Expression level of TNF-α was significantly lower both in recovered COVID-19 patients and vaccinated women in comparison to healthy controls.

Functional ice creams were developed by adding larvae of the insect Tenebrio molitor mixed with a seed (Salvia hispanica) and a pseudocereal (Chenopodium quinoa) to strawberry–cranberry ice cream. The objective was to increase micronutrients, macronutrients, and antioxidants, thus rendering the product a food complement. Four ice cream formulations were manufactured: the control strawberry–cranberry ice cream and three experimental mixtures, one of them with an addition of Tenebrio larvae (HT) and two others with a combination of Tenebrio larvae, chia (HTC), and quinoa (HTQ). The ice creams were submitted to proximate chemical analysis: mineral, fatty acid, vitamin, and one antioxidant (cyanidin 3 glucoside) determination. The strawberry–cranberry ice cream was used as a control formulation to evaluate if there were significant differences among nutrients, to which a Dunnett test with a critical value of α = 0.05% was applied. The three formulations that were studied showed a significant increase in the analyzed micronutrients and macronutrients compared to the control formulation. We observed increases of up to 62% in lipid content in the HTC formulation, while an increase of 41% in the protein content of the HT formulation was observed. We quantified an increase and enrichment of vitamins and minerals in the manufactured products, so that their nutritional value was significantly enhanced. In the determination of cyanidin 3 glucoside, we found that the formulation to which chia had been added showed an increase of 74% as compared to the control ice cream; this is important because anthocyanins are a group of flavonoids that stand out for their antioxidant and antimutagenic capabilities.

Abstract Although a protocol for immediate postoperative thirst management has been applied clinically in a discrete way, most surgical and nursing teams remain neglected, unlike other complications in the postoperative period. This pilot study aimed to demonstrate if the ice popsicles promote thirst satiety and subsequently investigate the mechanism involved in this process. Twenty-five volunteers in the immediate postoperative period were recruited to participate in this study conducted in a tertiary medical center in southern Brazil. In this protocol, ice popsicles decreased the intensity and discomfort of thirst. Relevantly, all patients (100%) demonstrated satiety after ice popsicles administrated in the immediate postoperative period. Altogether, ice popsicles constitute a promising, innovative, and simple strategy for the management of thirst, mainly after surgery, adding to patient comfort and the security of the health team that manages it.

1,2:3,5-Di-O-isopropylidene-α-D-xylofuranose (DX) is a major component of a new bio-crude: a viscous oil presenting petroleum-friendly properties produced by the ketalization of sugarcane bagasse. This article studies DX HDO (hydrodeoxygenation) over a Pd/HBEA catalyst in a batch reactor at 250°C. The effects of hydrogen pressure from 10 to 40 bar, catalyst/DX ratio from ½ to 2, and reaction time 0–24 h were investigated. A range of conditions for complete hydrodeoxygenated DX into alkanes with a Pd/HBEA catalyst was found. In these conditions, a low coke yield with water as the principal deoxygenated product was obtained. Further, higher amounts of alkanes containing seven or more carbons (A7+) were favored at 30 bar of hydrogen pressure, Cat/DX ratio = 2, and short reaction time. Products analysis that accompanied the above variations during reaction time led to general insights into reaction pathways. First, in the presence of DX, an effective n-hexane conversion was not observed on experiments of low catalyst/DX ratio (½) or in the initial period of high Cat/DX ratio, suggesting DX is much more successful than n-hexane to compete for active sites. Then, the formation of a pool of oxygenated compounds, such as furans, ketones, and carboxylic acids, along with lighter and heavier alkanes was observed. Hence, the aforementioned oxygenates may undergo reactions, such as aldol condensation with subsequent hydrodeoxygenation reaction, generating heavier alkanes.

Nickel (Ni) leftovers arise from both catalyst application interventions and Ni alloy piping of the cooking oil industry (COI) being wasted as pollutants of freshwater bodies via discharged effluent. The current study assessed one of the indigenously feasible Ni removal systems comprising autochthonous Arthrospira platensis Gomont (AP)-driven Ni phycoremediation cells (NPCs). After screening AP for hyperaccumulation in the Ni spiked solution, AP was transferred to the NPCs. Propagation of the AP inoculum was proportionate to the pollution load drop of COI with 22.97 and 55.07% drops in the biochemical (BOD) and chemical oxygen demand (COD), respectively. With the 0.11 bioconcentration factor, there was an uptake of 14.24 g mineral with 16.22% Ni removal and a 36.35 desorption ratio. The experimental data closely fitted with the Langmuir and Freundlich isotherms, respectively. The study concluded that A. platensis could be taken for treatment of Ni-loaded industrial effluents at the microcosmic level.

Due to the fast growth of cities and populations, residential areas are increasingly close to Waste Water Treatment Plants (WWTPs), causing odour impact on neighbourhood and complaints. The present study focuses on assessing the impact on the residential vicinity of a WWTP in Valencia (Spain) of different superficial odour sources using Computational Fluid Dynamics (CFD) simulations. Together with other software, CFD simulation is used as a complementary study to model pollutant dispersion. Unlike other simulation tools, CFD is bounded to smaller domains (1 or 2km). However, it performs more meaningful calculations and provides more detailed results, which allow obtaining resolutions at a scale below centimetre. The precise mesh generated in this work facilitates the study of the effect of the turbulence and the eddies caused by different obstacles (buildings, walls, etc.).

Macrophages metabolic reprogramming in response to microbial insults is a major determinant of pathogen growth or containment. Here, we reveal a distinct mechanism by which stimulator of interferon genes (STING), a cytosolic sensor that regulates innate immune responses, contributes to an inflammatory M1-like macrophage profile upon Brucella abortus infection. This metabolic reprogramming is induced by STING-dependent stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), a global regulator of cellular metabolism and innate immune cell functions. HIF-1α stabilization reduces oxidative phosphorylation and increases glycolysis during infection with B. abortus and, likewise, enhances nitric oxide production, inflammasome activation and IL-1β release in infected macrophages. Furthermore, the induction of this inflammatory profile participates in the control of bacterial replication since absence of HIF-1α renders mice more susceptible to B. abortus infection. Mechanistically, activation of STING by B. abortus infection drives the production of mitochondrial reactive oxygen species (mROS) that ultimately influences HIF-1α stabilization. Moreover, STING increases the intracellular succinate concentration in infected macrophages, and succinate pretreatment induces HIF-1α stabilization and IL-1β release independently of its cognate receptor GPR91. Collectively, these data demonstrate a pivotal mechanism in the immunometabolic regulation of macrophages during B. abortus infection that is orchestrated by STING via HIF-1α pathway and highlight the metabolic reprogramming of macrophages as a potential treatment strategy for bacterial infections.

Objective To evaluate whether the addition of colchicine to standard treatment for COVID-19 results in better outcomes.Design We present the results of a randomised, double-blinded, placebo-controlled clinical trial of colchicine for the treatment of moderate to severe COVID-19, with 75 patients allocated 1:1 from 11 April to 30 August 2020. Colchicine regimen was 0.5 mg thrice daily for 5 days, then 0.5 mg twice daily for 5 days. The primary endpoints were the need for supplemental oxygen, time of hospitalisation, need for admission and length of stay in intensive care unit and death rate.Results Seventy-two patients (36 for placebo and 36 for colchicine) completed the study. Median (and IQR) time of need for supplemental oxygen was 4.0 (2.0–6.0) days for the colchicine group and 6.5 (4.0–9.0) days for the placebo group (p

This research encompasses the use of noxious weed Parthenium hysterophorus as feedstock for pyrolysis carried out at varying temperatures of 300, 450 and 600°C. Temperature significantly affected the yield and properties of the pyrolysis products including char, syngas and bio-oil. Biochar yield decreased from 61% to 37% from 300 °C to 600 °C, whereas yield of gas and oil increased with increasing temperature. The pyrolysis products were physico-chemically characterized. In biochar, pH, conductivity, fixed carbon, ash content, bulk density and specific surface area of the biochar increased whereas cation exchange capacity, calorific value, volatile matter, hydrogen, nitrogen and oxygen content decreased with increasing temperature. Thermogravimetric analysis showed that the biochar prepared at higher temperature was more stable. Gas Chromatography-Mass Spectrometry analysis of biochar indicated the presence of alkanes, alkenes, nitriles, fatty acids, esters, amides and aromatic compounds. Number of compounds decreased with increasing temperature, but aromatic compounds increased with increasing temperature. Scanning electron micrographs of biochar prepared at different temperatures indicated micropore formation at lower temperature while increase in the size of pores and disorganization of vessels occurred at increasing temperature. The chemical composition was found to be richer at lower pyrolysis temperature. GC–MS analysis of the bio-oil indicated the presence of phenols, ketones, acids, alkanes, alkenes, nitrogenated compounds, heterocyclics and benzene derivatives.

Brucella abortus is a Gram-negative bacterium responsible for a worldwide zoonotic infection-Brucellosis, which has been associated with high morbidity rate in humans and severe economic losses in infected livestock. The natural route of infection is through oral and nasal mucosa but the invasion process through host gut mucosa is yet to be understood. Studies have examined the role of NLRP6 (NOD-like receptor family pyrin domain-containing-6 protein) in gut homeostasis and defense against pathogens. Here, we investigated the impact of gut microbiota and NLRP6 in a murine model of Ba oral infection. Nlrp6-/- and wild-type (WT) mice were infected by oral gavage with Ba and tissues samples were collected at different time points. Our results suggest that Ba oral infection leads to significant alterations in gut microbiota. Moreover, Nlrp6-/- mice were more resistant to infection, with decreased CFU in the liver and reduction in gut permeability when compared to the control group. Fecal microbiota transplantation from WT and Nlrp6-/- into germ-free mice reflected the gut permeability phenotype from the donors. Additionally, depletion of gut microbiota by broad-spectrum-antibiotic treatment prevented Ba replication in WT while favoring bacterial growth in Nlrp6-/-. Finally, we observed higher eosinophils in the gut and leukocytes in the blood of infected Nlrp6-/- compared to WT-infected mice, which might be associated to the Nlrp6-/- resistance phenotype. Altogether, these results indicated that gut microbiota composition is the major factor involved in the initial stages of pathogen host replication and partially also by the resistance phenotype observed in Nlrp6 -/- mice regulating host inflammation against Ba infection.

The Schistosoma mansoni SmKI-1 protein is composed of two domains: a Kunitz-type serine protease inhibitor motif (KD) and a C-terminus domain with no similarity outside the genera. Our previous work has demonstrated that KD plays an essential role in neutrophil elastase (NE) binding blockage, in neutrophil influx and as a potential anti-inflammatory molecule. In order to enhance NE blocking capacity, we analyzed the KD sequence from a structure-function point of view and designed specific point mutations in order to enhance NE affinity. We substituted the P1 site residue at the reactive site for a leucine (termed RL-KD), given its central role for KD's inhibition to NE. We have also substituted a glutamic acid that strongly interacts with the P1 residue for an alanine, to help KD to be buried on NE S1 site (termed EA-KD). KD and the mutant proteins were evaluated in silico by molecular docking to human NE, expressed in Escherichia coli and tested towards its NE inhibitory activity. Both mutated proteins presented enhanced NE inhibitory activity in vitro and RL-KD presented the best performance. We further tested RL-KD in vivo in an experimental model of monosodium urate (MSU)-induced acute arthritis. RL-KD showed reduced numbers of total cells and neutrophils in the mouse knee cavity when compared to KD. Nevertheless, both RL-KD and KD reduced mice hypernociception in a similar fashion. In summary, our results demonstrated that both mutated proteins showed enhanced NE inhibitory activity in vitro. However, RL-KD had a prominent effect in diminishing inflammatory parameters in vivo.

Solar distillation system depends on adhesion of water molecules inside the glass cover of solar stills. Regular glass surfaces are prone to adsorbing other unwanted compounds and in turn lower the percentage of recovery of pure distilled water. In this study, the production of purified distilled water was compared with and without the use of graphene-based surface modifiers. In areas where salt content is high, the first pass is still usually laden with salts. Hence, to improve adhesion of water and rejection of salts, both the inside glass cover surfaces and the metal absorber plates were modified using oxygen plasma treatment and graphene surface enhancement. Results showed a 48.9% improvement of distilled water recovery from an initial recovery of 2.90 L/m2 per day to an average of 4.32 L/m2 per day. In addition, the resulting distilled water passes the World Health Organization drinking water standards such as pH, electrical conductivity (EC), and salinity. The average reduction in EC was 96.52%, an average increase of 5.06% of pH, and an average reduction of salinity of 96.52%, all measured at the highest brine salinity of 5%. The reported value of EC was 23.33 μS/cm, a lowest and near-neutral pH of 6.85, and an average salinity of 12.10 ppm.

COVID-19 is a worldwide emergency; therefore, there is a critical need for foundational knowledge about B and T cell responses to SARS-CoV-2 essential for vaccine development. However, little information is available defining which determinants of SARS-CoV-2 other than the spike glycoprotein are recognized by the host immune system. In this study, we focus on the SARS-CoV-2 nucleocapsid protein as a suitable candidate target for vaccine formulations. Major B and T cell epitopes of the SARS-CoV-2 N protein are predicted and resulting sequences compared with the homolog immunological domains of other coronaviruses that infect human beings. The most dominant of B cell epitope is located between 176–206 amino acids in the SRGGSQASSRSSSRSRNSSRNSTPGSSRGTS sequence. Further, we identify sequences which are predicted to bind multiple common MHC I and MHC II alleles. Most notably there is a region of potential T cell cross-reactivity within the SARS-CoV-2 N protein position 102–110 amino acids that traverses multiple human alpha and betacoronaviruses. Vaccination strategies designed to target these conserved epitope regions could generate immune responses that are cross-reactive across human coronaviruses, with potential to protect or modulate disease. Finally, these predictions can facilitate effective vaccine design against this high priority virus.