Synthesis of cannabidiol-based compounds as ACE2 inhibitors with potential application in the treatment of COVID-19.

Cannabis is a general name for plants of the genus Cannabis. Used as fiber, medicine, drug, for religious, therapeutic, and hedonistic purposes along the millenia, it is mostly known for its psychoactive properties. One of its major constituents, cannabidiol (CBD), a non-psychoactive substance, among many other biological activities, has shown potential as an anti-SARS-CoV-2 drug. In this work, three derivatives and an analogue of CBD were synthesized, and cell viability and antiviral activities were evaluated. None of the compounds showed cytotoxicity up to a maximum concentration of 100 µM and, in contrast, displayed a significant antiviral activity, superior to remdesivir and nafamostat mesylate, with IC50 values ranging from 9.4 to 1.9 µM. In order to search for a possible molecular target, the inhibitory activity of the compounds against ACE2 was investigated, with expressive results (IC50 ranging from 3.96 µM to 0.01 µM).

Growth performance, hematology parameters and Aeromonas hydrophila challenge of pirarucu (Arapaima gigas) in response to dietary protein levels.

The pirarucu (Arapaima gigas) is a fast-growing Amazonian species of high commercial value. The present study aimed to determine the dietary crude protein (CP) level to promote maximum zootechnical performance for pirarucu fingerlings and as their resistance to Aeromonas hydrophila, as well as evaluate their hematological parameters. Pirarucu fingerlings (2.4 ± 0.08 g, 6.8 ± 0.52 cm) were distributed in 18 tanks (140 L, 40 fish per tank, n = 3) and fed six experimental diets consisting of increasing levels of CP: 300, 400, 450, 500, 550 and 650 g kg-1 in a completely randomized design. Data were submitted to one-way ANOVA (p < 0.05) and the ideal CP level for weight gain was determined using polynomial regression analysis. The dietary CP levels were evaluated using a quadratic polynomial regression and the level of 595 g kg-1 was determined for the best weight gain. The hematocrit of fish fed 300 g kg-1 was higher than in the other groups. No mortalities were observed after the 15-day bacterial challenge; however, number of pirarucu with bacterial damage on the pirarucu caudal fin was higher in the group that was fed the diet with 300 g kg-1. A dietary protein level of 618 g kg-1 is therefore recommended for providing maximum weight gain and immunological resistance in pirarucu fingerlings weighing 2.4-112.5 g.

Taming the SARS-CoV-2-mediated proinflammatory response with BromAc®.

Introduction: In the present study, the impact of BromAc®, a specific combination of bromelain and acetylcysteine, on the SARS-CoV-2-specific inflammatory response was evaluated. Methods: An in vitro stimulation system was standardized using blood samples from 9 healthy donors, luminex assays and flow cytometry were performed. Results and discussion: BromAc® demonstrated robust anti-inflammatory activity in human peripheral blood cells upon SARS-CoV-2 viral stimuli, reducing the cytokine storm, composed of chemokines, growth factors, and proinflammatory and regulatory cytokines produced after short-term in vitro culture with the inactivated virus (iSARS-CoV-2). A combined reduction in vascular endothelial growth factor (VEGF) induced by SARS-CoV-2, in addition to steady-state levels of platelet recruitment-associated growth factor-PDGFbb, was observed, indicating that BromAc® may be important to reduce thromboembolism in COVID-19. The immunophenotypic analysis of the impact of BromAc® on leukocytes upon viral stimuli showed that BromAc® was able to downmodulate the populations of CD16+ neutrophils and CD14+ monocytes observed after stimulation with iSARS-CoV-2. Conversely, BromAc® treatment increased steady-state HLA-DR expression in CD14+ monocytes and preserved this activation marker in this subset upon iSARS-CoV-2 stimuli, indicating improved monocyte activation upon BromAc® treatment. Additionally, BromAc® downmodulated the iSARS-CoV-2-induced production of TNF-a by the CD19+ B-cells. System biology approaches, utilizing comprehensive correlation matrices and networks, showed distinct patterns of connectivity in groups treated with BromAc®, suggesting loss of connections promoted by the compound and by iSARS-CoV-2 stimuli. Negative correlations amongst proinflammatory axis and other soluble and cellular factors were observed in the iSARS-CoV-2 group treated with BromAc® as compared to the untreated group, demonstrating that BromAc® disengages proinflammatory responses and their interactions with other soluble factors and the axis orchestrated by SARS-CoV-2. Conclusion: These results give new insights into the mechanisms for the robust anti-inflammatory effect of BromAc® in the steady state and SARS-CoV-2-specific immune leukocyte responses, indicating its potential as a therapeutic strategy for COVID-19.

Optimization of bioprocess of Schleiferilactobacillus harbinensis Ca12 and its viability in frozen Brazilian berries (Açai, Euterpe oleracea Mart.).

Amazonian palm berries (açaí, Euterpe oleracea Mart.) are fruits with high nutritional value and antioxidant activity and have aroused the interest of consumers, popularizing fruit pulps enriched with probiotics. Amazonian palm berries (açaí, Euterpe oleracea Mart.) are fruits with high nutritional potential, providing a source of carbohydrates, fibers, proteins, lipids, vitamins, and minerals. Furthermore, açai provides several health benefits, including antioxidant activity. Nutritionally enhanced foods have aroused the interest of consumers, popularizing fruit pulps enriched with probiotics. Probiotics are dietary supplements consisting of live, beneficial microorganisms in the host which improve the intestinal microbiota. The objective of this study was to isolate, identify, and characterize the probiotic potential of an isolated Schleiferilactobacillus harbinensis strain (dubbed Ca12) and provide an optimized bioprocess for its production, using the complete factorial and central rotational compound design to supplement the frozen açai pulp. The isolated strain S. harbinensis Ca12 presented adequate resistance to gastric juice and bile salts, microbial activity against different Candida strains, self-aggregation and coaggregation properties, high adhesion in HT-29 cells, and 35% inhibition of Salmonella in HT-29 cells. When optimized, the cellular biomass production of the S. harbinensis Ca12 strain was approximately 600% higher than the unsupplemented whey, with a production of 3.6 × 1010 CFU mL-1. The S. harbinensis Ca12 strain's viability in the creamy and traditional frozen açai pulp was shown to be stable for up to 6 months at 20 °C. The impact of this study involved for the first time the S. harbinensis Ca12 described in the Brazilian cocoa pulp with activity against Candida albicans of clinical importance, creating the potential of a new functional food with important benefits to human health as prevention for candidiasis.

Effect of EDTA enriched diets on farmed fish allergenicity and muscle quality; a proteomics approach.

Fish is one of the most common elicitors of food-allergic reactions worldwide. These reactions are triggered by the calcium-binding muscle protein ß-parvalbumin, which was shown to have reduced immunoglobulin E (IgE)-binding capacity upon calcium depletion. This work aimed to reduce gilthead seabream allergenicity using diets supplemented with a calcium chelator. Three experimental feeds were tested, differing in ethylenediaminetetraacetic acid (EDTA) supplementation, and its effects on muscle and parvalbumin's IgE-reactivity were analyzed. Chromatographic determination of EDTA showed no accumulation in the muscle and sensory results demonstrated that the lowest concentration did not affect fish quality as edible fish. Proteomics revealed one protein related to muscle contraction with significantly different relative abundance. Immunoblot assays performed with fish-allergic patients sera indicated a 50% reduction in IgE-reactivity upon EDTA presence. These preliminary results provide the basis for the further development of a non-GMO approach to modulate fish allergenicity and improve safety of aquaculture fish.

Dietary Creatine Supplementation in Gilthead Seabream (Sparus aurata): Comparative Proteomics Analysis on Fish Allergens, Muscle Quality, and Liver.

The quality of fish flesh depends on the skeletal muscle's energetic state and delaying energy depletion through diets supplementation could contribute to the preservation of muscle's quality traits and modulation of fish allergens. Food allergies represent a serious public health problem worldwide with fish being one of the top eight more allergenic foods. Parvalbumins, have been identified as the main fish allergen. In this study, we attempted to produce a low allergenic farmed fish with improved muscle quality in controlled artificial conditions by supplementing a commercial fish diet with different creatine percentages. The supplementation of fish diets with specific nutrients, aimed at reducing the expression of parvalbumin, can be considered of higher interest and beneficial in terms of food safety and human health. The effects of these supplemented diets on fish growth, physiological stress, fish muscle status, and parvalbumin modulation were investigated. Data from zootechnical parameters were used to evaluate fish growth, food conversion ratios and hepatosomatic index. Physiological stress responses were assessed by measuring cortisol releases and muscle quality analyzed by rigor mortis and pH. Parvalbumin, creatine, and glycogen concentrations in muscle were also determined. Comparative proteomics was used to look into changes in muscle and liver tissues at protein level. Our results suggest that the supplementation of commercial fish diets with creatine does not affect farmed fish productivity parameters, or either muscle quality. Additionally, the effect of higher concentrations of creatine supplementation revealed a minor influence in fish physiological welfare. Differences at the proteome level were detected among fish fed with different diets. Differential muscle proteins expression was identified as tropomyosins, beta enolase, and creatine kinase among others, whether in liver several proteins involved in the immune system, cellular processes, stress, and inflammation response were modulated. Regarding parvalbumin modulation, the tested creatine percentages added to the commercial diet had also no effect in the expression of this protein. The use of proteomics tools showed to be sensitive to infer about changes of the underlying molecular mechanisms regarding fish responses to external stimulus, providing a holistic and unbiased view on fish allergens and muscle quality.