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Welfare metrics have been established for octopuses in the laboratory, but not for octopuses living in the wild. Wild octopuses are constantly exposed to potentially harmful situations, and the ability to assess the welfare status of wild octopuses could provide pertinent information about individuals’ health and species’ resilience to stressors. Here, we used underwater photos and videos to identify injuries and stress-related behaviors in wild Octopus insularis in a variety of contexts, including interacting with fishermen, interacting with other octopuses and fish, proximity to predators, in den, foraging, and in senescence. We adapted established metrics of octopus welfare from the laboratory to these wild octopuses. In addition to observing all of the stress measures, we also identified two previously unknown measures associated with decreased welfare: (1) a half white eye flash and (2) a half-and-half blotch body pattern. More than half of the individuals analyzed had arm loss, and almost half of the individuals had skin injuries. We also observed that irregular chromatophore expression and abnormal motor coordination were associated with interactions with fishermen. This is the first study to apply measures of welfare from the laboratory to wild octopuses. Our results may also aid in the identification of welfare measures for other wild invertebrates.

Background Mesenchymal stem cells (MSCs) are promising for cell-based therapies targeting a wide range of diseases. However, challenges in translating MSC-based therapies to clinical applications necessitate standardized protocols following Good Manufacturing Practices (GMP) guidelines. This study aimed at developing GMP-complained protocols for FPMSCs isolation and manipulation, necessary for translational research, by (1) optimize culture of MSCs derived from an infrapatellar fat pad (FPMSC) condition through animal-free media comparison and (2) establish feasibility of MSC isolation, manufacturing and storage under GMP-compliance (GMP-FPMSC). Methods FPMSCs from three different patients were isolated following established protocols and the efficacy of two animal component-free media formulations in the culturing media were evaluated. The impact of different media formulations on cell proliferation, purity, and potency of MSCs was evaluated through doubling time, colony forming unit assay, and percentage of MSCs, respectively. Furthermore, the isolation and expansion of GMP-FPMSCs from four additional donors were optimized and characterized at each stage according to GMP requirements. Viability and sterility were checked using Trypan Blue and Bact/Alert, respectively, while purity and identity were confirmed using Endotoxin, Mycoplasma assays, and Flow Cytometry. The study also included stability assessments post-thaw and viability assessment to determine the shelf-life of the final GMP-FPMSC product. Statistical analyses were conducted using one-way ANOVA with Tukey’s Multiple Comparisons. Results The study demonstrated that FPMSCs exhibited enhanced proliferation rates when cultured in MSC-Brew GMP Medium compared to standard MSC media. Cells cultured in this media showed lower doubling times across passages, indicating increased proliferation. Additionally, higher colony formation in FPMSCs cultured in MSC-Brew GMP Medium were observed, supporting enhanced potency. Data from our GMP validation, including cells from 4 different donors, showed post-thaw GMP-FPMSC maintained stem cell marker expression and all the specifications required for product release, including > 95% viability (> 70% is required) and sterility, even after extended storage (up to 180 days), demonstrating the reproducibility and potential of GMP-FPMSCs for clinical use as well as the robustness of the isolation and storage protocols. Conclusions The study underscores the feasibility of FPMSCs for clinical uses under GMP conditions and emphasizes the importance of optimized culture protocols to improve cell proliferation and potency in MSC-based therapies.

The present study aimed to evaluate saline extracts from the leaves (LE) and stem (SE) of Portulaca elatior in relation to their phytochemical composition and photoprotective and antioxidant effects, as well as to evaluate the toxicity of the leaf extract. The extracts were characterized for protein concentration and phenol and flavonoid contents, as well as for thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) profiles. Total antioxidant capacity and DPPH and ABTS + scavenging activities were determined. In the photoprotective activity assay, the sun protection factor (SPF) was calculated. The toxicity evaluation of LE included in vitro hemolytic assay and in vivo oral and dermal acute toxicity assays in Swiss mice. LE showed the highest protein, phenol, and flavonoid (8.79 mg/mL, 323.46 mg GAE/g, and 101.96 QE/g, respectively). TLC revealed the presence of flavonoids, reducing sugars, terpenes, and steroids in both extracts. In HPLC profiles, LE contained flavonoids, while SE contained flavonoids and ellagic tannins. The antioxidant activity assays showed the lowest IC 50 values ​(34.15–413.3 µg/mL) for LE, which presented relevant SPF (> 6) at 50 and 100 µg/mL. LE demonstrated low hemolytic capacity, and no signs of intoxication were observed in mice treated orally or topically at 1000 mg/kg. However, at 2000 mg/kg, an increase in the mean corpuscular volume of erythrocytes and a reduction in lymphocytes were observed; animals treated topically with 2000 mg/kg displayed scratching behavior during the first hour of observation and showed edema and erythema that regressed after six days. In conclusion, LE did not present acute oral or dermal toxicity in Swiss mice at a dose of 1000 mg/kg and showed slight toxicity in animals treated with 2000 mg/kg.

The co-digestion of swine manure with vegetable waste is an alternative that can increase the production of biogas and methane generated by the isolated digestion of manure. However, recommendations that are based on the best ratio between manure and forage, as well as the age of harvest, are still scarce in the literature. This study was conducted to evaluate inclusions (0, 25, 50, 75 and 100%) of the total solids (TS) of Elephant grass (Pennisetum purpureum Schum) harvested at two ages medium age (MA) at 45 days of growth and advanced age (AA) at 90 days in co-digestion with swine manure, using an entirely randomized design in a 5x2 factorial scheme. Batch digesters were used and biogas production was monitored for 12 weeks. There was influence of forage age (P <0.05) on the degradation of solids and neutral detergent fiber, with higher values for the substrates containing MA forage. The highest CH4 yields were obtained by the substrates containing MA forage in the inclusion of 27.7 and 31.6%, being 253.7 and 222.2 L of CH4 per Kg of total or volatile solids. The age of the forages influenced the onset and persistence of biogas production, being advantageous only in the inclusion of 25% of MA forage. The AA forage inclusion is not recommended for co-digestion with swine manure. RESUMO: A co-digestão dos dejetos suínos associados a resíduos vegetais é uma alternativa que pode elevar as produções de biogás e metano alcançadas pela digestão isolada do dejeto. No entanto, recomendações que se baseiem na melhor proporção entre dejeto e forragem, assim como na idade de colheita ainda são escassas na literatura. O objetivo do trabalho foi avaliar inclusões crescentes (0, 25, 50, 75 e 100% dos sólidos totais (ST) de capim elefante (Pennisetum purpureum Schum.) colhido em duas idades de corte mediana (MA) aos 45 dias de crescimento e avançada (AA) aos 90 dias, empregado em co-digestão com o dejeto suíno, adotando-se delineamento inteiramente ao acaso, em esquema fatorial 5x2. Foram utilizados biodigestores batelada de bancada, sendo as produções de biogás acompanhadas durante 12 semanas. Houve influência da idade da forragem (P<0,05) na degradação de sólidos e fibra em detergente neutro, com maiores valores para os substratos que continham a forragem MA. Os maiores rendimentos de CH4 foram alcançados pelos substratos que continham forragem MA na inclusão de 27,7 e 31,6 %, sendo de 253,7 e 222,2 L de CH4 por Kg de ST ou sólido volátil adicionado. A idade das forragens influenciou o início e a persistência das produções de biogás ao longo do período de digestão, sendo vantajosa apenas na inclusão de 25% de forragem com MA.

Studies have reported that Croton blanchetianus Baill., Euphorbiaceae, has anti-inflammatory effects triggered by the presence of polyphenols. With the absence of studies directed to obtain optimized extracts, the objective of this work was to obtain and characterize a spray-dried extract of C. blanchetianus leaves using different statistical tools. The extracts were optimized by a simplex centroid by analysis of residue, total polyphenol, total flavonoid, and rutin content, besides the fingerprints by high-performance thin layer and high-performance liquid chromatography. The spray-dried extract was obtained with maltodextrin/colloidal silicon dioxide. A factorial design evaluated the drying conditions with factors temperature (°C) and flow rate (l/h), and the responses were moisture content, yield, total flavonoid, total polyphenol, rutin content, and rheological properties. Higher total flavonoid and better response of dry residue and total polyphenol were observed with the water:acetone mixture. Regarding the drying process, the maltodextrin did not show interference, obtaining extract with higher polyphenol content, better thermal stability, and flow properties. It was possible to obtain a dry extract with better characteristics using 0.5 l/h and 150 °C, without affecting the chemical profile. These results establish that C. blanchetianus can be an alternative source of flavonoids which infers its potential in pharmaceutical products. Graphical Abstract

Millions of plastic surgeries are performed worldwide every year with the objective of correcting lipodystrophies stemming from lesions, tumor resections, birth defects, and AIDS-associated antiretroviral therapy. Besides that, a large number of clinical research have assessed the outcome of procedures that rely on combinations of dermal fillers and autologous cells. However, little is known about the safety of these combinations and the localization of the injected cells. The aim of this study was to test the toxicity of a solution containing 1% hyaluronic acid (HA) and adipose-derived stromal cells (ASCs) from the human adipose tissue and to assess the localization of the injected cells, with and without HA, labeled with technetium-99m. Rats received subcutaneous and intraperitoneal injections of a solution containing 1% HA/adipose-derived stromal cells isolated from the human fat tissue. The animals were then observed for up to forty-two days. The solution tested in this study did not result in systemic, biochemical, or anatomic alterations that could represent toxicity symptoms. The association of HA and ASCs labeled with technetium-99m remained at the site of the injection within a period of twenty-four hours, as demonstrated by a whole-body imaging software fusion of SPECT and CT. In conclusion, our study shows that the subcutaneous and intraperitoneal injection of HA associated with adipose-derived stromal cells (ASCs) is safe. The association of HA and ASCs did not induce local or systemic toxicity. Thus, the administration of volume equal to or less than 0.2 mL of the agent filler (1×106 ASC+HA 1%) should be considered for subsequent studies and may be an alternative to dermal fillers due to the expected lasting effects.

A injeção de carvão pulverizado (PCI) é uma técnica utilizada em altos-fornos (AFs) para reduzir o consumo de coque. A queima do carvão pulverizado injetado nas ventaneiras do AF ocorre em pressões médias (até 450 kPa), temperaturas de chama elevadas (cerca de 2500 K), taxas de aquecimento muito elevadas (105 –106 K/s) e tempos de residência muito curtos, abaixo de 40 ms. Como não há testes padronizados para avaliação da combustibilidade de carvões nas condições de PCI, simuladores de combustão em PCI são comumente utilizados para esse fim. Este trabalho apresenta as características relevantes do simulador de PCI desenvolvido pelo LASID-UFRGS, que opera com pulso automatizado de injeção. As características principais do simulador são a medida direta de pressão e temperatura dentro da zona de combustão durante o, a medição de temperatura com termopares ultrarrápidos (ms), a aquisição de dados e controle em alta velocidade (ms) e a coleta dos produtos sólidos e gasosos da combustão. Os primeiros resultados mostraram a evolução da pressão e temperatura durante o pulso de injeção, com ou sem amostra, o grau de conversão (burnout) do carvão e a composição dos gases de combustão para diferentes carvões. A análise estatística dos dados experimentais demonstrou uma boa repetitividade para todos os carvões testados.

The polyethylene (PE) and polypropylene (PP) microplastics (MPs) can act as carriers of the molecule 3-tertiary-butyl-4-hydroxyanisole (3BHA), which propose harmful impacts to aqueous ecosystems. Meanwhile, 3BHA has already been detected in several environments and human urine and serum samples. 3BHA is an antioxidant in foods, food packaging, cosmetics and pharmaceuticals. However, it raised concerns about its possible endocrine-disrupting effect in recent years. The interaction between two MPs and 3BHA could start in farmland and be maintained during transportation to the ocean. Herein, the adsorption behavior and mechanism of 3BHA by PE and PP MPs were investigated via molecular dynamics (MD) simulation, density functional theory (DFT), non-covalent interactions (NCI), the total density of states (TDOS) and frontier molecular orbital (FMO). Furthermore, the stability of 3BHA adsorbed complexes was investigated by adsorption-free energies (Δ Gads ), showing that the 3BHA has a significant interaction with the MPs studied, with energies of -1.31 kcal mol −1 and − 4.19 kcal mol −1 for PE and PP, respectively. Intermolecular van der Waals forces were one of the primary adsorption mechanisms of 3BHA by MPs, as evidenced by NCI calculations. It was demonstrated that the adsorption of the 3BHA in MPs decreases the energy gap ( E g ) of the HOMO and LUMO orbitals; for PE MP, the E g decreases from 12.8386 to 9.1445 eV, and from 12.7957 to 9.2092 eV for PP MP. The adsorption of the 3BHA on the MPs resulted in an increase in the TDOS distribution and a slight shift in the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) states. In conclusion, this study about the mechanism of adsorption of the 3BHA in PE and PP MPs can provide new evidence and enhance our understanding of the environmental behavior of 3BHA in the environment.

The co-digestion of swine manure with vegetable waste is an alternative that can increase the production of biogas and methane generated by the isolated digestion of manure. However, recommendations that are based on the best ratio between manure and forage, as well as the age of harvest, are still scarce in the literature. This study was conducted to evaluate inclusions (0, 25, 50, 75 and 100%) of the total solids (TS) of Elephant grass (Pennisetum purpureum Schum) harvested at two ages medium age (MA) at 45 days of growth and advanced age (AA) at 90 days in co-digestion with swine manure, using an entirely randomized design in a 5x2 factorial scheme. Batch digesters were used and biogas production was monitored for 12 weeks. There was influence offorage age (P <0.05) on the degradation of solids and neutral detergent fiber, with higher values for the substrates containing MA forage. The highest C[H.sub.4] yields were obtained by the substrates containing MA forage in the inclusion of 27.7 and 31.6%, being 253.7 and 222.2 L of CH4 per Kg of total or volatile solids. The age of the forages influenced the onset and persistence of biogas production, being advantageous only in the inclusion of 25% of MA forage. The AA forage inclusion is not recommended for co-digestion with swine manure. Key words: biogas, fiber degradation, methane, solids reductions. A co-digestão dos dejetos suínos associados a resíduos vegetais é uma alternativa que pode elevar as produções de biogás e metano alcançadas pela digestão isolada do dejeto. No entanto, recomendações que se baseiem na melhor proporção entre dejeto e forragem, assim como na idade de colheita ainda são escassas na literatura. O objetivo do trabalho foi avaliar inclusões crescentes (0, 25, 50, 75 e 100% dos sólidos totais (ST) de capim elefante (Pennisetum purpureum Schum.) colhido em duas idades de corte mediana (MA) aos 45 dias de crescimento e avançada (AA) aos 90 dias, empregado em co-digestão com o dejeto suíno, adotando-se delineamento inteiramente ao acaso, em esquema fatorial 5x2. Foram utilizados biodigestores batelada de bancada, sendo as produções de biogás acompanhadas durante 12 semanas. Houve influência da idade da forragem (P<0,05) na degradação de sólidos e fibra em detergente neutro, com maiores valores para os substratos que continham a forragem MA. Os maiores rendimentos de C[H.sub.4] foram alcançados pelos substratos que continham forragem MA na inclusão de 27,7 e 31,6 %, sendo de 253,7 e 222,2 L de C[H.sub.4] por Kg de ST ou sólido volátil adicionado. A idade das forragens influenciou o início e a persistência das produções de biogás ao longo do período de digestão, sendo vantajosa apenas na inclusão de 25% de forragem com MA. Palavras-chave: biogás, degradação de fibra, metano, redução de sólidos.

This study aims to characterize the waste of weaned piglets and estimate the emissions of N2O in kg of CO2 eq/kg of weight gain in the first week of housing. Primary data were obtained in the first week after weaning of piglets to identify how much waste from this animal category may affect the environment. The life cycle assessment was applied to verify the amount of manure and the emission of nitrous oxide (N2O) considering the weight gain (WG) of piglets in the first post-weaning week. Eight waste collections were carried out in two lots representing an average of 8,099 animals with initial and final weight of 5.01 and 5.84 kg, respectively. The production of residues was 0.128 kg of dry matter (DM) for each kilogram of WG produced. This waste production has an emission capacity of approximately 4x10-4 kg N2O/kg WG in the first post-weaning week. Considering that N2O has a global warming potential almost 300 times higher in retaining heat than CO2, each 1 kg of piglet produced can emit about 0.129 kg of CO2 equivalent from the N2O produced. According to the number of piglets evaluated in this study, the total emission can reach 1.85 tons of CO2 equivalent in the first post-weaning week alone.