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Nine mature Katahdin wethers (70 ± 1.5 kg initial BW) were used in a crossover experiment to evaluate effects on digestion and energy utilization of levels of feed intake being used in studies addressing the maintenance energy requirement with limited nutrient intake. A 50% concentrate pelleted diet composed of 20.0% ground alfalfa, 29.1% cottonseed hulls, 9.0% cottonseed meal, 20.0% ground corn, 13.0% wheat middlings, 5.0% pelleting agent, and 3.9% other ingredients was fed near the ME requirement for maintenance (Control; 44.4 g/kg BW0.75 DMI) and at 55% of this level (Restricted; 24.4 g/kg BW0.75 DMI). Periods were 4 wk in length, with 3 wk for adaptation, measures in the final week when situated in metabolism cages, and 2 d for gas exchange measurement via a head-box respiration calorimetry system. Apparent total tract digestibilities of DM (65.8 and 73.9% [SEM 1.92]), OM (76.3 and 81.7% [SEM 1.48]), CP (72.1 and 78.5% [SEM 1.70]), NDF (32.3 and 49.0% [SEM 3.34]), and GE (64.7 and 73.0% [SEM 1.97]) were greater (P < 0.05) for Restricted intake than for Control intake. Expressed in megajoules per day, quantities of energy in urine (0.94 and 0.72 [SEM 0.320]) and ruminally emitted methane (1.02 and 0.76 [SEM 0.085]) were greater for Control vs. Restricted intake (P < 0.05), but as a percentage of DE, they tended to be greater for Restricted intake (8.1 and 10.8% [SEM 3.52; P = 0.056] in urine and 9.0 and 11.1% [SEM 0.75; P = 0.096] in methane for Control and Restricted intake, respectively). As a consequence, ME intake as a percentage of GE intake did not differ (P = 0.301) between treatments (53.5 and 57.3% [SEM 2.88] for Control and Restricted intake, respectively). The difference in heat energy (447 and 379 [SEM 15.1]) was less than that in ME intake (395 and 225 kJ/kg BW0.75 [SEM 21.4] for Control and Restricted intake, respectively). In conclusion, restricted feed intake had marked influences on digestibility, although effect on metabolizability was tempered by changes in urinary and methane energy, the former presumably impacted by lean tissue mobilization. Based on the magnitude of difference between ME intake and heat energy with restricted intake, lower heat energy and less tissue mobilization would be expected with longer periods. [ABSTRACT FROM AUTHOR]

Thirty-seven Dorper (DOR), 35 Katahdin (KAT), and 31 St. Croix (STC) ewes (57, 58, and 44 kg [SEM 2.2]) from 45 commercial farms in the Midwest (MW), Northwest (NW), Southeast (SE), and central Texas (TX), between 2.2 and 3.4 yr of age, were used to evaluate responses to high heat load index (HLI) conditions. There were 4 sequential 2-wk periods (8 wk total) with target HLI during day/nighttime of 70/70, 85/70, 90/77, and 95/81, with weekly measures at 0700 (before increased daytime HLI), 1300, and 1700 h (preceding lower nighttime HLI). Rectal temperature (RT; °C) was affected (P = 0.003) by breed × time (38.58, 38.92, and 39.07 for DOR, 38.67, 38.92, and 39.05 for KAT, and 38.45, 38.69, and 38.85 for STC at 0700, 1300, and 1700 h, respectively [SEM 0.034]). There were interactions between week and time (P < 0.001) in respiration rate (RR; breaths/min; 52, 72, 66, and 85 at 0700 h; 120, 130, 151, and 144 at 1300 h; and 116, 123, 141, and 142 at 1700 h [SEM 3.1]) and panting score (0-4; 0.05, 0.03, 0.11, and 0.28 at 0700 h; 0.48, 0.86, 1.61, and 1.47 at 1300 h; and 0.76, 0.91, 1.54, and 1.51 at 1700 h in wk 5, 6, 7, and 8, respectively [SEM 0.042]). Breed × time RR (P = 0.008) means were 57, 107, and 103 for DOR, 55, 101, and 96 for KAT, and 47, 88, and 90 for STC at 0700, 1300, and 1700 h, respectively (SEM 3.1); however, there was an interaction (P = 0.007) among breed, region, and time (57, 110, and 101 for MW DOR; 59, 110, and 108 for MW KAT; 43, 89, and 88 for MW STC; 65, 113, and 111 for NW DOR; 54, 104, and 96 for NW KAT; 56, 92, and 94 for NW STC; 49, 93, and 96 for SE DOR; 52, 105, and 96 for SE KAT; 45, 79, and 87 for SE STC; 57, 110, and 104 for TX DOR; 54, 83, and 84 for TX KAT; and 46, 91, and 89 for TX STC at 0700, 1300, and 1700 h, respectively [SEM 6.1]). In conclusion, RT of STC was low at all times compared with DOR and KAT, even with lower RR. There appeared to be considerable adaptation from wk 1 to 2 during the 2 highest HLI periods via evening respiration. Region effects varied with breed, such as relatively high RR by STC from the NW to maintain low RT, lower RR of DOR from the SE than other regions, and a smaller difference among times in RR of KAT from TX. [ABSTRACT FROM AUTHOR]

Twenty-four Dorper (DOR; 60 ± 2.8 kg initial BW), 23 Katahdin (KAT; 63 ± 2.6 kg BW), and 21 St. Croix (STC; 40 ± 2.7 kg BW) female sheep (0.9-8.9 yr) from 45 commercial farms in the Midwest, Northwest, Southeast, and central Texas were used to evaluate resilience to water restriction. Animals were housed and fed a 15% CP, 50% concentrate pelleted diet at 160% of the ME requirement for maintenance. In period 1, 2 wk in duration, ad libitum water intake was determined followed by 25 and 50% decreases in water availability in periods 2 and 3, which were 2 and 5 wk in duration, respectively. Water was offered at 0700 h and blood was sampled at 0800 and 1400 h on the last 2 d of each week. Data presented are means of values within periods, with breed and region means separated by LSD and linear and quadratic contrasts addressing period effects. There were no effects of or interactions involving region for the variables analyzed. During the baseline period, water intake was 3.59, 3.79, and 3.00 kg/d for DOR, KAT, and STC, respectively (SEM 0.140), and DMI averaged 58.6 ± 0.98 g/kg BW0.75. There were linear decreases (P < 0.01) in DMI from period 1 to 3 of similar magnitude for DOR and KAT (134 and 153 g/d, respectively), whereas there was no change for STC (P = 0.52; 27 g/d [pooled SEM 41.5]). Plasma osmolality (mOsmol/kg) was 301, 307, and 303 for DOR; 302, 307, and 305 for KAT; and 306, 308, and 307 for STC in periods 1, 2, and 3, respectively (SEM 1.4), with a quadratic effect of advancing period for DOR (P < 0.01) and linear and quadratic effects (P < 0.05) for KAT. There were no effects on packed cell volume (P > 0.05). Serum protein concentration quadratically changed (P < 0.001) as period advanced for each breed (7.1, 6.0, and 7.2 g/dL in periods 1, 2, and 3, respectively [SEM 0.11]). In conclusion, there were no indications of influences of region on resilience to water restriction based on period means, and the lack of change in DMI by STC with limited water availability suggests relatively high resilience for this breed. [ABSTRACT FROM AUTHOR]

Thirty-seven Dorper, 35 Katahdin, and 31 St. Croix ewes (57, 58, and 44 kg [SEM 2.2]) from 45 commercial farms in the Midwest (MW), Northwest (NW), Southeast (SE), and central Texas (TX), between 2.2 and 3.4 yr of age, were used to evaluate responses to high heat load index (HLI) conditions. There were 4 sequential 2-wk periods with target HLI during day/nighttime of 70/70, 85/70, 90/77, and 95/81. A 15% CP and 50% concentrate pelleted diet was fed at 120% of the ME requirement for maintenance, and water was offered free choice. Body weight was measured 3 times each week, and blood was sampled at 1300 h on the last day of each period. There was an interaction (P < 0.001) between period and week within period in BW, with slightly greater values in wk 2 vs. 1 of periods 3 and 4 and a greater difference between period 1 and 4 values in wk 2 than 1 (53.1, 54.1, 54.9, and 55.4 kg in wk 1, and 53.0, 54.2, 55.4, and 56.1 kg in wk 2 in periods 1, 2, 3, and 4, respectively [SEM 0.85]). There was an interaction (P = 0.037) in DMI (g/kg BW0.75) among region, period, and week, with values generally similar between weeks in periods 1 and 2 relative to those in periods 3 and 4 (51.0, 52.4, 51.0, and 51.2 in period 3 and wk 1; 49.5, 52.1, 50.8, and 51.6 in period 3 and wk 2; 49.6, 52.1, 50.6, and 49.4 in period 4 and wk 1; and 48.9, 52.0, 49.7, and 46.3 in period 4 and wk 2 for MW, NW, SE, and TX, respectively [SEM 1.09]). Neither blood glucose nor lactate concentration was affected by breed (P > 0.05), but there were breed differences (P < 0.02) in serum concentrations of creatinine (0.91, 0.81, and 0.77 mg/dL [SEM 0.023]), total protein (6.13, 6.42, and 6.81 g/dL [SEM 0.156]), and urea N (17.4, 18.0, and 20.0 mg/dL [SEM 0.54] for Dorper, Katahdin, and St. Croix, respectively). In conclusion, some blood constituent levels suggest breed differences in resilience to high HLI. Differences among periods and weeks in BW presumably relate to increased water consumption with high HLI. The interaction in DMI may reflect differences among regions in rate of adaptation to high HLI and the contribution of decreased feed intake to coping with high HLI. [ABSTRACT FROM AUTHOR]

Fumarate hydratase (FH) mutated papillary renal cell carcinoma is an aggressive variant of kidney cancer that poorly responds to conventional targeted therapies and immunotherapy. Here, we present the 10-year follow-up of a heavily pre-treated patient with several lines of therapy, achieving a remarkable complete response to anti-PD-1 rechallenge. In addition, we highlight a common immune-related adverse event of anti-PD-1, eosinophilia, as a possible biomarker of response and using TCGA data analysis, provide proof-of-concept for tumor expression of the eosinophil-related gene SIGLEC8, as a promising powerful predictor of prognosis for papillary renal cell carcinoma patients., (© 2024. The Author(s).)

Introduction: Nanoparticles (NPs) are widely used in the pharmaceutical field to treat various human disorders. Among these, lipid-based NPs (LNPs), including solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), are favored for drug/bioactive delivery due to their high stability, biocompatibility, encapsulation efficiency, and sustained/controlled release. These properties make them particularly suitable as carriers of compounds derived from plant sources., Areas Covered: This study comprehensively explores updated literature knowledge on SLN and NLC, focusing on their composition and production methods for the specific delivery of drug/bioactive compounds derived from plant sources of interest in pharmaceutical and biomedical fields., Expert Opinion: SLN and NLC facilitate the development of more effective natural product-based therapies, aiming to reduce dosage and minimize side effects. These delivery systems align with the consumer demands for safer and more sustainable products, as there are also based on biocompatible and biodegradable raw materials, thereby posing minimal toxicological risks while also meeting regulatory guidelines.

Introduction: Numerous purified bioactive compounds, crude extracts, and essential oils have demonstrated potent antioxidant, antimicrobial, anti-inflammatory, and antiviral properties, particularly in vitro or in silico; however, their in vivo applications are hindered by inadequate absorption and distribution in the organism. The incorporation of these phytochemicals into solid lipid nanoparticles (SLN) or nanostructured lipid carriers (NLC) has demonstrated significant advancements and represents a viable approach to improve their bioavailability through different administration routes., Areas Covered: This review discusses the potential applications of SLN and NLC, loading bioactive compounds sourced from plants for the treatment of several diseases. An overview of the preclinical developments on the use of these lipid nanoparticles is also provided as well as the requisites to be launched on the market., Expert Opinion: Medicinal plants have gained even more value for the pharmaceutical industries and their customers, leading to many studies exploring their therapeutic potential. Several bioactives derived from plants with antiviral, anticancer, neuroprotective, antioxidant, and antiaging properties have been proposed and loaded into lipid nanoparticles. In vitro and invivo studies corroborate the added value of SLN/NLC to improve the bioavailability of several bioactives. Surface modification to increase their stability and target delivery should be considering.

Analysis on spatial-temporal data has several benefits that range from an improved traffic network in a city to increased earnings for drivers and ridesharing companies. A common analysis technique is clustering. However, most clustering techniques consider a constrained representation of clusters that is limited to a single timestamp. Evolving clusters exist through several timestamps and interact with other spatial-temporal objects or clusters, having cluster relationships. When many evolving clusters exist for analysis, a graph can be used to represent cluster evolution. In this article, we propose a framework for graph-based cluster evolution representation and analysis. The framework represents cluster structure and relationships as well as provides a graph representation of cluster evolution for analysis. Evaluation is done in three case studies with spatial-temporal data about taxis that can identify important phenomena or trends in movements in a city for traffic improvement., (© 2024. The Author(s).)

Lyme disease (LD) is a globally distributed zoonotic multisystemic condition caused by gram-negative spirochete bacteria of the Borrelia burgdorferi complex, transmitted through tick bites. Research on LD in domestic animals in Portugal is limited, potentially leading to underestimating its prevalence. This disease affects many species, including humans, making it a critical public health issue. In domestic animals, LD often presents subclinically or with non-specific clinical signs, complicating its diagnosis. Nevertheless, veterinarians should always consider LD in cases with a history of tick exposure and compatible clinical signs. Diagnostic confirmation can be achieved through serological and other complementary tests. Treatment involves eradicating the bacterial infection and managing clinical signs using a combination of antibiotics, analgesics, anti-inflammatories, and other medications. Effective prevention primarily relies on tick control measures. This review aims to provide an up-to-date state-of-the-art LD, particularly in Portugal., (© 2024. The Author(s).)

Parenteral administration is one of the most commonly used drug delivery routes for nanoparticle-based dosage forms, such as lipid-based and polymeric nanoparticles. For the treatment of various diseases, parenteral administration include intravenous, subcutaneous, and intramuscular route. In drug development phase, multiparameter strategy with a focus on drug physicochemical properties and the specificity of the administration route is required. Nanoparticle properties in terms of size and targeted delivery, among others, are able to surpass many drawbacks of conventional dosage forms, but these unique properties can be a bottleneck for approval by regulatory authorities. Quality by Design (QbD) approach has been widely utilized in development of parenteral nanoparticle-based dosage forms. It fosters knowledge of product and process quality by involving sound scientific data and risk assessment strategies. A full and comprehensive investigation into the state of implementation and applications of the QbD approach in these complex drug products can highlight the gaps and challenges. In this review, the analysis of critical attributes and Design of Experiment (DoE) approach in different nanoparticulate systems, together with the proper utilization of Process Analytical Technology (PAT) applications are described. The essential of QbD approach for the design and development of nanoparticle-based dosage forms for delivery via parenteral routes is discussed thoroughly., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Natural killer (NK) cells have gained attention as a promising adoptive cell therapy platform for their potential to improve cancer treatments. NK cells offer distinct advantages over T-cells, including major histocompatibility complex class I (MHC-I)-independent tumor recognition and low risk of toxicity, even in an allogeneic setting. Despite this tremendous potential, challenges persist, such as limited in vivo persistence, reduced tumor infiltration, and low absolute NK cell numbers. This review outlines several strategies aiming to overcome these challenges. The developed strategies include optimizing NK cell expansion methods and improving NK cell antitumor responses by cytokine stimulation and genetic manipulations. Using K562 cells expressing membrane IL-15 or IL-21 with or without additional activating ligands like 4-1BBL allows "massive" NK cell expansion and makes multiple cell dosing and "off-the-shelf" efforts feasible. Further improvements in NK cell function can be reached by inducing memory-like NK cells, developing chimeric antigen receptor (CAR)-NK cells, or isolating NK-cell-based tumor-infiltrating lymphocytes (TILs). Memory-like NK cells demonstrate higher in vivo persistence and cytotoxicity, with early clinical trials demonstrating safety and promising efficacy. Recent trials using CAR-NK cells have also demonstrated a lack of any major toxicity, including cytokine release syndrome, and, yet, promising clinical activity. Recent data support that the presence of TIL-NK cells is associated with improved overall patient survival in different types of solid tumors such as head and neck, colorectal, breast, and gastric carcinomas, among the most significant. In conclusion, this review presents insights into the diverse strategies available for NK cell expansion, including the roles played by various cytokines, feeder cells, and culture material in influencing the activation phenotype, telomere length, and cytotoxic potential of expanded NK cells. Notably, genetically modified K562 cells have demonstrated significant efficacy in promoting NK cell expansion. Furthermore, culturing NK cells with IL-2 and IL-15 has been shown to improve expansion rates, while the presence of IL-12 and IL-21 has been linked to enhanced cytotoxic function. Overall, this review provides an overview of NK cell expansion methodologies, highlighting the current landscape of clinical trials and the key advancements to enhance NK-cell-based adoptive cell therapy.

While N-acetyl azaaurones have already been disclosed for their potential against tuberculosis (TB), their low metabolic stability remains an unaddressed liability. We now report a study designed to improve the metabolic stability and solubility of the azaaurone scaffold and to identify the structural requirements for antimycobacterial activity. Replacing the N-acetyl moiety for a N-carbamoyl group led to analogues with sub- and nanomolar potencies against M. tuberculosis H37Rv, as well as equipotent against drug-susceptible and drug-resistant M. tuberculosis isolates. The new N-carbamoyl azaaurones exhibited improved microsomal stability, compared to their N-acetylated counterparts, with several compounds displaying moderate to high kinetic solubility. The frequency of spontaneous resistance to azaaurones was observed to be in the range of 10 -8 , a value that is comparable to current TB drugs in the market. Overall, these results reveal that azaaurones are amenable to structural modifications to improve metabolic and solubility liabilities, and highlight their potential as antimycobacterial agents., (© 2023 Wiley-VCH GmbH.)

Successful implementation of adoptive cell therapy (ACT) of cancer requires comprehensively addressing biological and practical challenges. This approach has been largely overlooked, resulting in a gap between the potential of ACT and its actual effectiveness. We summarize the most promising technical strategies in creating an "ideal" ACT product, focusing on chimeric antigen receptor (CAR)-engineered cells. Since many requirements for effective ACT are common to most cancers, what we outline here might have a broader impact., Competing Interests: Declaration of interests Q.C., S.W., V.P., and D.B. are employees and shareholders of Kite Pharma. L.S.Q. is a founder of Epic Bio and a scientific advisor of Laboratory of Genomics Research (LGR) and Kytopen. T.K.L. is a co-founder of Senti Biosciences, Synlogic, Engine Biosciences, Tango Therapeutics, Corvium, BiomX, Eligo Biosciences, Bota.Bio, NE47Bio, Protuoso, and Raina Bio. T.K.L. also holds financial interests in nest.bio, Armata, IndieBio, CognitoHealth, Quark Biosciences, Personal Genomics, Thryve, Lexent Bio, MitoLab, Vulcan, Serotiny, Pulmobiotics, Provectus Algae, Invaio, NSG Biolabs, PairX, NSG Ventures, Integrated Biosciences, NextBiotics, Flagship VL43, Resvita Bio, and Absci. M.J.L. is a founder, shareholder, and employee of Outpace Bio, as well as a founder and shareholder of Lyell Immunopharma. M.J.L. is an inventor on patents licensed by Outpace Bio. F.M. is an employee and options holder of Sonata Therapeutics, Watertown, MA., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) strains are of particular concern, especially strains with mobilizable carbapenemase genes such as blaKPC, blaNDM or blaOXA-48, given that carbapenems are usually the last line drugs in the β-lactam class and, resistance to this sub-class is associated with increased mortality and frequently co-occurs with resistance to other antimicrobial classes., Objectives: To characterize the genomic diversity and international dissemination of CRKP strains from tertiary care hospitals in Lisbon, Portugal., Methods: Twenty CRKP isolates obtained from different patients were subjected to WGS for species confirmation, typing, drug resistance gene detection and phylogenetic reconstruction. Two additional genomic datasets were included for comparative purposes: 26 isolates (ST13, ST17 and ST231) from our collection and 64 internationally available genomic assemblies (ST13)., Results: By imposing a 21 SNP cut-off on pairwise comparisons we identified two genomic clusters (GCs): ST13/GC1 (n = 11), all bearing blaKPC-3, and ST17/GC2 (n = 4) harbouring blaOXA-181 and blaCTX-M-15 genes. The inclusion of the additional datasets allowed the expansion of GC1/ST13/KPC-3 to 23 isolates, all exclusively from Portugal, France and the Netherlands. The phylogenetic tree reinforced the importance of the GC1/KPC-3-producing clones along with their rapid emergence and expansion across these countries. The data obtained suggest that the ST13 branch emerged over a decade ago and only more recently did it underpin a stronger pulse of transmission in the studied population., Conclusions: This study identifies an emerging OXA-181/ST17-producing strain in Portugal and highlights the ongoing international dissemination of a KPC-3/ST13-producing clone from Portugal., (© The Author(s) 2023. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Molecular dynamics simulations employing the all-atom optimized potential for liquid simulations (OPLS-AA) force field were performed for determining self-diffusion coefficients (D11) of ethanol and tracer diffusion coefficients (D12) of solutes in ethanol at several temperature and pressure conditions. For simulations employing the original OPLS-AA diameter of ethanol's oxygen atom (σOH), calculated and experimental diffusivities of protic solutes differed by more than 25%. To correct this behavior, the σOH was reoptimized using the experimental D12 of quercetin and of gallic acid in liquid ethanol as benchmarks. A substantial improvement of the calculated diffusivities was found by changing σOH from its original value (0.312 nm) to 0.306 nm, with average absolute relative deviations (AARD) of 3.71% and 4.59% for quercetin and gallic acid, respectively. The new σOH value was further tested by computing D12 of ibuprofen and butan-1-ol in liquid ethanol with AARDs of 1.55% and 4.81%, respectively. A significant improvement was also obtained for the D11 of ethanol with AARD = 3.51%. It was also demonstrated that in the case of diffusion coefficients of non-polar solutes in ethanol, the original σOH=0.312 nm should be used for better agreement with experiment. If equilibrium properties such as enthalpy of vaporization and density are estimated, the original diameter should be once again adopted.

SARS-CoV-2 infection and its clinical manifestations (COVID-19) quickly evolved to a pandemic and a global public health emergency. The limited effectivity of available treatments aimed at reducing virus replication and the lessons learned from other coronavirus infections (SARS-CoV-1 or NL63) that share the internalization process of SARS-CoV-2, led us to revisit the COVID-19 pathogenesis and potential treatments. Virus protein S binds to the angiotensin-converting enzyme 2 (ACE2) initiating the internalization process. Endosome formation removes ACE2 from the cellular membrane preventing its counter-regulative effect mediated by the metabolism of angiotensin II to angiotensin (1-7). Internalized virus-ACE2 complexes have been identified for these coronaviruses. SARS-CoV-2 presents the highest affinity for ACE2 and produces the most severe symptoms. Assuming ACE2 internalization is the trigger for COVID-19 pathogenesis, accumulation of angiotensin II can be viewed as the potential cause of symptoms. Angiotensin II is a strong vasoconstrictor, but has also important roles in hypertrophy, inflammation, remodeling, and apoptosis. Higher levels of ACE2 in the lungs explain the acute respiratory distress syndrome as primary symptoms. Most of the described findings and clinical manifestations of COVID-19, including increased interleukin levels, endothelial inflammation, hypercoagulability, myocarditis, dysgeusia, inflammatory neuropathies, epileptic seizures and memory disorders can be explained by excessive angiotensin II levels. Several meta-analyses have demonstrated that previous use of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers were associated with better prognosis for COVID-19. Therefore, pragmatic trials to assess the potential therapeutic benefits of renin-angiotensin-aldosterone system inhibitors should be urgently promoted by health authorities to widen the therapeutic options for COVID-19., (Copyright © 2023 Sociedade Portuguesa de Cardiologia. Publicado por Elsevier España, S.L.U. All rights reserved.)

To evaluate the genetic diversity and clustering rates of M. tuberculosis strains to better understand transmission among persons deprived of liberty (PDL) in Rio Grande do Sul (RS), southern Brazil. This is a cross-sectional study, including strains of M. tuberculosis isolated from PDL, stored at the Central Laboratory of RS, in the period from 2013 to 2018. The molecular characterization was performed using the MIRU-VNTR 15 loci method. A total of 598 M. tuberculosis strains were genotyped, and 37.5% were grouped into 53 clusters. Cluster sizes ranged from 2 to 34 strains. The largest cluster of the study had strains from 34 PDL, and 58.8% of the PDL of this cluster were in P01. Among the clusters formed, in 60.3%, there was at least one strain from P01. The most common strains in RS were LAM (53.2%) and Haarlem (31.1%). The LAM strain was the most likely to form clusters, and Haarlem was associated with anti-TB drug resistance. This was translational research, and the results can collaborate with the TB control programs, leading to improved strategies that allow the reduction of the TB burden in prisons., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In this work, a simple two-parameters correlation based on the Rice and Gray, Lennard-Jones, and Stockmayer theories was devised for the calculation of binary diffusion coefficients (D12) of any type of solutes at infinite dilution in polar and non-polar solvents. This equation can be relevant for systems with polar solvents, since most models in the literature fail when strong intermolecular forces predominate in solution. The new correlation embodies the Stockmayer potential without requiring the dipole moments of any component, which significantly enlarges its application. It was validated with the largest D12 database of polar and non-polar dense systems, with 8812 data points (NDP) spanning 553 systems, of which 133 have water as solvent (NDP = 1266), 89 contain polar solvents excluding water (NDP = 1405), 177 have supercritical carbon dioxide (SC-CO2) as solvent (NDP = 5028), and 154 have non-polar or weakly polar solvents excluding SC-CO2 (NDP = 1113). Overall, the model achieved an average deviation of only 3.43%, with accurate and unbiased behavior even for polar systems.

We performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain Mycobacterium tuberculosis H37Rv. The most active compound 11 was used for in vitro selection of spontaneous resistant mutants. The same mutations in six genes were detected in bacterial cultures grown under increased concentrations of 11 (2×, 4×, 8× MIC). The mutant positions were presented as mixed wild type and mutant alleles while increasing the concentration of the compound led to the semi-proportional and significant increase in mutant alleles. The identified genes belong to different categories and pathways. Some of them were previously reported as mediating drug resistance or drug tolerance, and counteracting oxidative and nitrosative stress, in particular: Rv0224c , fbiC , iniA , and Rv1592c . Gene-set interaction analysis revealed a certain weak interaction for gene pairs Rv1592-Rv1639c and Rv1592-Rv0224c . To conclude, this study experimentally demonstrated a multifaceted primary genetic response of M. tuberculosis to the action of nitrofurans. All three 11 -treated subcultures independently presented the same six SNPs, which suggests their non-random occurrence and likely causative relationship between compound action and possible resistance mechanism.

The main aim of this study was to describe the clinical and immunological outcomes, as well as the inflammatory profile, of patients with advanced HIV in an assisted-living facility in which an outbreak of SARS-CoV-2 occurred. SARS-CoV-2 humoral and specific T-cell response were analyzed in patients with HIV infection and COVID-19; as a secondary objective of the analysis, levels of the inflammatory markers (IL-1β, IL-6, IL-8, and TNFα) were tested in the HIV/COVID-19 group, in HIV-positive patients without COVID-19, and in HIV-negative patients with mild/moderate COVID-19. Antibody kinetics and ability to neutralize SARS-CoV-2 were evaluated by ELISA assay, as well as the inflammatory cytokines; SARS-CoV-2 specific T-cell response was quantified by ELISpot assay. Mann−Whitney or Kruskal−Wallis tests were used for comparisons. Thirty patients were included with the following demographics: age, 57 years old (IQR, 53−62); 76% male; median HIV duration of infection, 18 years (15−29); nadir of CD4, 57/mmc (23−100) current CD4 count, 348/mmc (186−565). Furthermore, 83% had at least one comorbidity. The severity of COVID-19 was mild/moderate, and the overall mortality rate was 10% (3/30). Additionally, 90% of patients showed positive antibody titers and neutralizing activity, with a 100% positive SARS-CoV-2 specific T-cell response over time, suggesting the ability to induce an effective specific immunity. Significantly higher levels of IL-6, IL-8, and TNF-α in COVID-19 without HIV vs. HIV/COVID-19 patients (p < 0.05) were observed. HIV infection did not seem to negatively impact COVID-19-related inflammatory state and immunity. Further data are mandatory to evaluate the persistence of these immunity and its ability to expand after exposure and/or vaccination.

The oral route is the best way to administer a drug; however, fitting peptide drugs in this route is a major challenge. In insulin cases, less than 0.5% of the administered dose achieves systemic circulation. Oral delivery by nanoparticles can increase insulin permeability across the intestinal epithelium while maintaining its structure and activity until release in the gut. This system can be improved to increase permeability across intestinal cells through active delivery. This study aimed to improve a nanoparticle formulation by promoting functionalization of its surface with immunoglobulin G to increase its absorption by intestinal epithelium. The characterization of formulations showed an adequate size and a good entrapment efficiency. Functionalized nanoparticles led to a desirable increase in insulin release time. Differential scanning calorimetry, infrared spectroscopy and paper chromatography proved the interactions of nanoparticle components. With immunoglobulin G, the nanoparticle size was slightly increased, which did not show aggregate formation. The developed functionalized nanoparticle formulation proved to be adequate to carry insulin and potentially increase its internalization by epithelial gut cells, being a promising alternative to the existing formulations for orally administered low-absorption peptides., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Increasing awareness regarding fossil fuel dependence, waste valorization, and greenhouse gas emissions have prompted the emergence of new solutions for numerous markets over the last decades. The tire industry is no exception to this, with a global production of more than 1.5 billion tires per year raising environmental concerns about their end-of-life recycling or disposal. Pyrolysis enables the recovery of both energy and material from end-of-life tires, yielding valuable gas, liquid, and solid fractions. The latter, known as recovered carbon black (rCB), has been extensively researched in the last few years to ensure its quality for market applications. These studies have shown that rCB quality depends on the feedstock composition and pyrolysis conditions such as type of reactor, temperature range, heating rate, and residence time. Recent developments of activation and demineralization techniques target the production of rCB with specific chemical, physical, and morphological properties for singular applications. The automotive industry, which is the highest consumer of carbon black, has set specific targets to incorporate recycled materials (such as rCB) following the principles of sustainability and a circular economy. This review summarizes the pyrolysis of end-of-life tires for the production of syngas, oil, and rCB, focusing on the process conditions and product yield and composition. A further analysis of the characteristics of the solid material is performed, including their influence on the rCB application as a substitute of commercial CB in the tire industry. Purification and modification post-treatment processes for rCB upgrading are also inspected.

Amazonas suffered greatly during the COVID-19 pandemic. The mortality and fatality rates soared and scarcity of oxygen and healthcare supplies led the health system and funerary services to collapse. Thus, we analyzed the trends of incidence, mortality, and lethality indicators of COVID-19 and the dynamics of their main determinants in the state of Amazonas from March 2020 to June 2021. This is a time-series ecological study. We calculated the lethality, mortality, and incidence rates with official and public data from the Health Department. We used the Prais-Winsten regression and trends were classified as stationary, increasing, or decreasing. The effective reproduction number (Rt) was also estimated. Differences were considered significant when p < 0.05. We extracted 396,772 cases of and 13,420 deaths from COVID-19; 66% of deaths were in people aged over 60; 57% were men. Cardiovascular diseases were the most common comorbidity (28.84%), followed by diabetes (25.35%). Rural areas reported 53% of the total cases and 31% of the total deaths. The impact of COVID-19 in the Amazon is not limited to the direct effects of the pandemic itself; it may present characteristics of a syndemic due to the interaction of COVID-19 with pre-existing illnesses, endemic diseases, and social vulnerabilities.

Safe and effective vaccines against coronavirus disease 2019 (COVID-19) are essential for ending the ongoing pandemic. Although impressive progress has been made with several COVID-19 vaccines already approved, it is clear that those developed so far cannot meet the global vaccine demand alone. We describe a COVID-19 vaccine based on a replication-defective gorilla adenovirus expressing the stabilized prefusion severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein named GRAd-COV2. We assessed the safety and immunogenicity of a single-dose regimen of this vaccine in healthy younger and older adults to select the appropriate dose for each age group. For this purpose, a phase 1, dose-escalation, open-labeled trial was conducted including 90 healthy participants (45 aged 18 to 55 years old and 45 aged 65 to 85 years old) who received a single intramuscular administration of GRAd-COV2 at three escalating doses. Local and systemic adverse reactions were mostly mild or moderate and of short duration, and no serious adverse events were reported. Four weeks after vaccination, seroconversion to spike protein and receptor binding domain was achieved in 43 of 44 young volunteers and in 45 of 45 older participants. Consistently, neutralizing antibodies were detected in 42 of 44 younger-age and 45 of 45 older-age volunteers. In addition, GRAd-COV2 induced a robust and T helper 1 cell (T H 1)–skewed T cell response against the spike protein in 89 of 90 participants from both age groups. Overall, the safety and immunogenicity data from the phase 1 trial support the further development of this vaccine.

Treatment of drug-resistant tuberculosis requires extended use of more toxic and less effective drugs and may result in retreatment cases due to failure, abandonment or disease recurrence. It is therefore important to understand the evolutionary process of drug resistance in Mycobacterium tuberculosis. We here in describe the microevolution of drug resistance in serial isolates from six previously treated patients. Drug resistance was initially investigated through phenotypic methods, followed by genotypic approaches. The use of whole-genome sequencing allowed the identification of mutations in the katG, rpsL and rpoB genes associated with drug resistance, including the detection of rare mutations in katG and mixed populations of strains. Molecular docking simulation studies of the impact of observed mutations on isoniazid binding were also performed. Whole-genome sequencing detected 266 single nucleotide polymorphisms between two isolates obtained from one patient, suggesting a case of exogenous reinfection. In conclusion, sequencing technologies can detect rare mutations related to drug resistance, identify subpopulations of resistant strains, and identify diverse populations of strains due to exogenous reinfection, thus improving tuberculosis control by guiding early implementation of appropriate clinical and therapeutic interventions., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Recurrent pulmonary tuberculosis (RPTB) is a growing, important and neglected problem affecting treated TB patients and TB health services across the world, particularly in sub-Saharan Africa. Analyses and identification of differences in clinical features between recurrent PTB and newly diagnosed PTB may lead to improved management recommendations., Methods: Between September 1 st 2019 and January 31 st 2020, we performed a prospective case controlled study of clinical and imaging features of patients with recurrent pulmonary tuberculosis and compared them with those of newly diagnosed PTB cases. Recurrent PTB was defined as a patient with bacteriologically confirmed active PTB who was previously successfully treated for PTB and was cured. A control was defined as a patient who presents for the first time with bacteriologically confirmed PTB. Clinical and radiological features were assessed and documented. Chi-square and t-test were used to test the difference between proportion and continuous data, respectively. Logistic regression analysis was done to determine factors associated with RPTB using SPSS version 23 software., Results: A total of 312 patients with PTB were enrolled (104 RPTB cases and 208 newly diagnosed controls). Clinically hemoptysis was more common in RPTB compared to controls 28/104 (26.9%) vs 35/208 (16.8%), P = 0.036. Chest pain was significantly less common among patients with RPTB compared to controls 33 (31.7%) vs 92 (44.2%), P = 0.034. A higher proportion of RPTB presented with cavitation 34/104 (32.7%) compared to control 44/208 (21.2%) P = 0.027. The median score for lung pathology was higher among patients with RPTB (50) compared to controls (30); P = 0.001. Lung function of patients with RPTB at diagnosis of index TB were more likely to show mixed restrictive and obstructive pattern 36/104 (34.6%) compared to controls 31/208 (14.9%). p<0.001. Multivariate analysis showed that patients older than 45 years of age (adjusted odds ratio [aOR]: 3.59, 95% CI: 1.38 - 9.32), those with hemoptysis (aOR 1.96, 95% CI: 1.04 - 3.69) p=0.04) and fibrosis on chest x rays (aOR 2.18, 95% CI: 1.16 - 4.10) were significantly associated with recurrent PTB., Conclusions: Hemoptysis, lung parenchymal damage, and patients being older than 45 years of age are significant features of RPTB. Management should focus on risk factors for recurrence, and a more holistic model of care to prevent long term lung injury., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Genomic-based surveillance on the occurrence of drug resistance and its transmission dynamics has emerged as a powerful tool for the control of tuberculosis (TB). A whole-genome sequencing approach, phenotypic testing and clinical-epidemiological investigation were used to undertake a retrospective population-based study on drug-resistant (DR)-TB in Rio Grande do Sul, the largest state in Southern Brazil. The analysis included 305 resistant Mycobacterium tuberculosis strains sampled statewide from 2011 to 2014, and covered 75.7% of all DR-TB cases identified in this period. Lineage 4 was found to be predominant (99.3%), with high sublineage-level diversity composed mainly of 4.3.4.2 [Latin American and Mediterranean (LAM)/RD174], 4.3.3 (LAM/RD115) and 4.1.2.1 (Haarlem/RD182) sublineages. Genomic diversity was also reflected in resistance of the variants to first-line drugs. A large number of distinct resistance-conferring mutations, including variants that have not been reported previously in any other setting worldwide, and 22 isoniazid-monoresistant strains with mutations described as disputed in the rpoB gene but causing rifampicin resistance generally missed by automated phenotypic tests as BACTEC MGIT. Using a cut-off of five single nucleotide polymorphisms, the estimated recent transmission rate was 55.1%, with 168 strains grouped into 28 genomic clusters. The most worrying fact concerns multi-drug-resistant (MDR) strains, of which 73.4% were clustered. Different resistance profiles and acquisition of novel mutations intraclusters revealed important amplification of resistance in the region. This study described the diversity of M. tuberculosis strains, the basis of drug resistance, and ongoing transmission dynamics across the largest state in Southern Brazil, stressing the urgent need for MDR-TB transmission control state-wide., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Tuberculosis (TB), caused by Mycobacterium tuberculosis, is one of the deadliest infectious diseases worldwide. Multidrug and extensively drug-resistant strains are making disease control difficult, and exhausting treatment options. New anti-TB drugs bedaquiline (BDQ), delamanid (DLM) and pretomanid (PTM) have been approved for the treatment of multi-drug resistant TB, but there is increasing resistance to them. Nine genetic loci strongly linked to resistance have been identified (mmpR5, atpE, and pepQ for BDQ; ddn, fgd1, fbiA, fbiB, fbiC, and fbiD for DLM/PTM). Here we investigated the genetic diversity of these loci across >33,000 M. tuberculosis isolates. In addition, epistatic mutations in mmpL5-mmpS5 as well as variants in ndh, implicated for DLM/PTM resistance in M. smegmatis, were explored. Our analysis revealed 1,227 variants across the nine genes, with the majority (78%) present in isolates collected prior to the roll-out of BDQ and DLM/PTM. We identified phylogenetically-related mutations, which are unlikely to be resistance associated, but also high-impact variants such as frameshifts (e.g. in mmpR5, ddn) with likely functional effects, as well as non-synonymous mutations predominantly in MDR-/XDR-TB strains with predicted protein destabilising effects. Overall, our work provides a comprehensive mutational catalogue for BDQ and DLM/PTM associated genes, which will assist with establishing associations with phenotypic resistance; thereby, improving the understanding of the causative mechanisms of resistance for these drugs, leading to better treatment outcomes., (© 2021. The Author(s).)

Photodynamic therapy is one of the more unique cancer treatment options available in today's arsenal against this devastating disease. It has historically been explored in cutaneous lesions due to the possibility of focal/specific effects and minimization of adverse events. Advances in drug delivery have mostly been based on biomaterials, such as liposomal and hybrid lipoidal vesicles, nanoemulsions, microneedling, and laser-assisted photosensitizer delivery systems. This review summarizes the most promising approaches to enhancing the photosensitizers' transdermal delivery efficacy for the photodynamic treatment for cutaneous pre-cancerous lesions and skin cancers. Additionally, discussions on strategies and advantages in these approaches, as well as summarized challenges, perspectives, and translational potential for future applications, will be discussed.