COVID-19 causes more severe and frequently fatal disease in patients with pre-existing comorbidities such as hypertension and heart disease. SARS-CoV-2 virus enters host cells through the angiotensin-converting enzyme 2 (ACE2), which is fundamental in maintaining arterial pressure through the renin-angiotensin system (RAS). Hypertensive patients commonly use medications such as angiotensin-converting enzyme inhibitors (ACEi), which can modulate the expression of ACE2 and, therefore, potentially impact the susceptibility and severity of SARS-CoV-2 infection. Here we assessed whether treatment of ACE2-humanized (K18-hACE2) mice with the ACEi Lisinopril affects lung ACE2 levels and the outcome of experimental COVID-19. K18-hACE2 mice were treated for 21 days with Lisinopril 10 mg/kg and were then infected with 105 PFU of SARS-CoV-2 (Wuhan strain). Body weight, clinical score, respiratory function, survival, lung ACE2 levels, viral load, lung histology, and cytokine (IL-6, IL-33, and TNF-α) levels were assessed. Mice treated with Lisinopril for 21 days showed increased levels of ACE2 in the lungs. Infection with SARS-CoV-2 led to massive decrease in lung ACE2 levels at 3 days post-infection (dpi) in treated and untreated animals, but Lisinopril-treated mice showed a fast recovery (5dpi) of ACE2 levels. Higher ACE2 levels in Lisinopril-treated mice led to remarkably higher lung viral loads at 3 and 6/7dpi. Lisinopril-treated mice showed decreased levels of the pro-inflammatory cytokines IL-6 and TNF-α in the serum and lungs at 6/7dpi. Marginal improvements in body weight, clinical score and survival were observed in Lisinopril-treated mice. No differences between treated and untreated infected mice were observed in respiratory function and lung histology. Lisinopril treatment showed both deleterious (higher viral loads) and beneficial (anti-inflammatory and probably anti-constrictory and anti-coagulant) effects in experimental COVID-19. These effects seem to compensate each other, resulting in marginal beneficial effects in terms of outcome for Lisinopril-treated animals.

The Zika Virus (ZIKV) is an emerging arbovirus of great public health concern, particularly in the Americas after its last outbreak in 2015. There are still major challenges regarding disease control, and there is no ZIKV vaccine currently approved for human use. Among many different vaccine platforms currently under study, the recombinant envelope protein from Zika Virus (rEZIKV) constitutes an alternative option for vaccine development and has great potential for monitoring ZIKV infection and antibody response. This study describes a method to obtain a bioactive and functional rEZIKV using an E. coli expression system, with the aid of a 5-L airlift bioreactor and following an automated fast protein liquid chromatography (FPLC) protocol, capable of obtaining high yields of approximately 20 mg of recombinant protein per liter of bacterium cultures. The purified rEZIKV presented preserved antigenicity and immunogenicity. Our results show that the use of an airlift bioreactor for the production of rEZIKV is ideal for establishing protocols and further research on ZIKV vaccines bioprocess, representing a promising system for the production of a ZIKV envelope recombinant protein-based vaccine candidate.

From the bubonic plague on the 14th century to the new Coronavirus disease 2019 (COVID-19), pandemics have profoundly changed societies function. Infectious disease outbreaks are getting shorter and shorter due to our densely populated cities, global travel, and nature mass exploration. In this regard, there is a particular concern about fires occurring in Brazil's Amazon rainforest, one of the most biodiverse places on earth that facilitates cross-species transmission giving rise to the emergency of new virulent pathogens. Situation is further complicated because Amazon spans across eight developing countries with limited preventive health care services. In this perspective, this review highlights the role of new methodologies best suited for epidemiological monitoring in low-income countries, such as high-throughput serological tests. Phage immunoprecipitation sequencing (Phip-Seq), for example, can evaluate antibody-repertoire binding specificities using oligonucleotides libraries encoding epitopes covering the DNA sequences from all human pathogenic viruses or all Arboviruses already described. After incubation with an individual's serum, these libraries can be immunoprecipitated for subsequent analysis by DNA sequencing. Data are analyzed revealing peptides recognized by the antibodies present in the sample. Being a technique at a relatively low cost, its implementation in developing countries is feasible and can generate very interesting scientific information.

Spike (S) protein has been recognized as a promising molecular target for diagnostic, vaccines and antiviral drugs development for COVID-19. In this study, we analyzed the most predominant mutations in the S protein of Brazilian isolates and predicted the effect of these amino acid alterations to protein conformation. A total of 25,924 sequences were obtained from GISAID for five regions of Brazilian territory (Midwest, North, Northeast, South, and Southeast), according to exclusion criteria. Most of the SARS-CoV-2 isolates belongs to the G clade and showed a large occurrence of D614G, N501Y and L18F substitutions. Prediction effects of these amino acid substitutions on the structure dynamics of the spike protein indicated a positive ΔΔG values and negative ΔΔSVib in most cases which is associated to structural stabilization and flexibility reduction of the S protein. Mutations E484K, N501Y and K417N belong to several SARS-CoV-2 variants of concern such as Alpha, Beta, Gamma and Delta, and showed high incidence among Brazilian isolates. These mutations have been described to increase RBD affinity to ACE-2 host and abolishment of RBD affinity to potent neutralizing ant-RBD. The increase in rates of infection and reinfection requires continuous genomic surveillance studies in order to characterize emerging mutations and monitor vaccine efficacy, and thus consideration structural data and dynamics in the observed phenotypes.Communicated by Ramaswamy H. Sarma.

The aim of this study was to analyze blood lactate response before, during and after simulated taekwondo fight in young male athletes. The experiment was composed of simulated official taekwondo fight. During the experiment a total of 7 blood samples (25 μL) were collected that following: at rest, after 1st, 2nd, and 3rd rounds, and 3, 5, and 10 min after the end of combat. The results showed that blood lactate concentrations [Lac]B increased significantly after 1st, 2nd, and 3rd rounds of the combat compared to the rested values. The [Lac]B after 3rd round was higher when compared to 3, 5, and 10 min of recovery. After 10 min of passive recovery, the [Lac]B was significantly lesser than 3 and 5 min of recovery, and 2nd round. These data showed that taekwondo requires high activation of anaerobic lactic metabolism, and improves the ability to rapidly remove the lactate from blood.

Hypontremia defined as a serum concentration below 135 mmol/L (1 mmol/L 1mEq/L). The aim of the present study was to analyze the effects of half-ironman triathlon on serum sodium concentration in Brazilian triathletes. Significant changes after triathlon completion was found pre = 141.72± 0.75 mmol/L; post= 138.37± 0.72 mmol/L (P=0.018). Was not found significant correlation between sodium plasma concentrations after the competition with the finishing time. In conclusion, the results presented herein, showing significative reduction in the sodium plasmatic concentrations remain between normal ranges.

Hypontremia defined as a serum concentration below 135 mmol/L (1 mmol/L - 1mEq/L). The aim of the present study was to analyze the effects of half-ironman triathlon on serum sodium concentration in Brazilian triathletes. Significant changes after triathlon completion was found pre = 141.72± 0.75 mmol/L; post= 138.37± 0.72 mmol/L (P=0.018). Was not found significant correlation between sodium plasma concentrations after the competition with the finishing time. In conclusion, the results presented herein, showing significative reduction in the sodium plasmatic concentrations remain between normal ranges.

Triathlon competition is dependent on the athletes’ ability to perform each discipline at optimal time, without excessive fatigue influencing the next one. Objectives: Determine the effects of a long distance triathlon on biochemistry parameters related to liver function. Design and methods: Blood samples from six athletes were collected before (T = 0) and immediately after the triathlon competition (T = 1). AST, ALT and alkaline phosphatase (ALP) values were assessed. Results: Significant changes after triathlon competition were found for AST and ALP and no significant changes were found for ALT over time.Conclusions: A series of metabolically alterations, mainly related to energy production and also to muscle and skeletal adaptations occurs during and after strenuous exercise. The altered status of those metabolical changes cannot directly reflect the intensity of any possible muscular or hepatic damage or overload and elevated AST/ALT ratio is better associated to skeletal muscle lesion during competition.

The freeze-dried aqueous extract (AE) from the aerial parts of Scoparia dulcis was tested for its effects on experimental gastric hypersecretion and ulcer in rodents. Administration of AE to animals with 4h pylorus ligature potently reduced the gastric secretion with ED50s of 195mg/kg (rats) and 306mg/kg (mice). The AE also inhibited the histamine- or bethanechol-stimulated gastric secretion in pylorus-ligated mice with similar potency suggesting inhibition of the proton pump. Bio-guided purification of the AE yielded a flavonoid-rich fraction (BuF), with a specific activity 4–8 times higher than the AE in the pylorus ligature model. BuF also inhibited the hydrolysis of ATP by H+,K+-ATPase with an IC50 of 500μg/ml, indicating that the inhibition of gastric acid secretion of Scoparia dulcis is related to the inhibition of the proton pump. Furthermore, the AE inhibited the establishment of acute gastric lesions induced in rats by indomethacin (ED50=313mg/kg, p.o.) and ethanol (ED50=490mg/kg, p.o.). No influence of the AE on gastrointestinal transit allowed discarding a possible CNS or a cholinergic interaction in the inhibition of gastric secretion by the AE. Collectively, the present data pharmacologically validates the popular use of Scoparia dulcis in gastric disturbances.

The effect of arginine (Arg) and Ornitargin® (OT) [a compound containing the aminoacids Arg, citrulline (Cit) and ornithine (Orn)] administration upon growth hormone (GH) gene expression was studied both in vivo and in vitro (hemipituitaries and GH3 cells) by Northern blot analysis. For in vivo studies, adult male Wistar rats were anesthetized, subjected to i.v. infusion of 200 µl of 150 mM NaCl (control group), Arg (15 or 150 mg) or OT (15 mg of Arg, 1 mg of Cit and 4 mg of Orn) at a rate of 20 µl/min, and killed 50 min thereafter. For the in vitro studies, hemipituitaries or GH3 cells were incubated in 1 ml of appropriate medium containing Arg (15 or 150 mg) or OT (15 mg of Arg, 1 mg of Cit and 4 mg of Orn) for 60 min. The pituitaries of the in vivo and in vitro studies and GH3 cells were subsequently processed for RNA extraction. Total RNA was subjected to electrophoresis in agarose (1%)/formaldehyde gel, transferred to a nylon membrane and subjected to hybridization with a rat GH 32P-cDNA, and 32P-18S rRNA probe to correct for the variability in RNA loading. After autoradiography of the membrane, the abundance of GH mRNA and 18S rRNA bands was quantified by densitometry. The in vivo study demonstrated that Arg and OT infusion induced a 2.3-fold increase in GH mRNA expression, which could result from the Arg-mediated inhibition of somatostatin release. In addition, in vitro Arg, but not OT, induced GH gene expression in hemipituitaries and GH3 cells, indicating that the aminoacid can act per se at the pituitary somatotrope level. In conclusion, our data show for the first time that arginine stimulates GH gene expression in parallel to its recognized GH-releasing activity.

The kinetic properties of the nicotinic receptor/ionic channel complex (AChR) were compared in cell cultures obtained from androgen-dependent skeletal muscles of the perineal complex (P) and from muscles less dependent upon sex hormones (the thigh musculature, T). Because the development of P is delayed compared to other skeletal muscles in the rat, cultures were performed taking into account the age of the donor (4- or 6-day-old rats), and the time interval the cells remained in culture (7 days and 15 days). The ionic channel conductance (γ) and the mean channel open time (τ) were determined with the patch-clamp technique in the cell-attached configuration at room temperature. Cultures from P and T muscles were morphologically identical in size and shape, independent of the animals' age at plating or on the plating time. In all of them, the AChR was spread over the cell membrane. More than one AChR ionic channel conductance was observed in P and T cultures, and the prevalent value of γ in either culture ranged from 30 pS to 35 pS. In P fibers from 4-day-old rats cultured for 7 days (P 4/7), the distribution of channel open times fitted a double exponential, while in T 4/7 they were fitted with a single exponential. In cultures from P and T muscles obtained from older rats (6 days old) and in those cells remaining in culture for a prolonged time (15 days), the channel open times also fitted a double exponential. Because P and T cultures lack trophic neuronal influences, the difference observed between the τ of P 4/7 and T 4/7 was thought to be the hormone requirement of P muscles to grow and differentiate. Likewise, the difference observed between T 4/7 and T 4/15 may indicate the need for neurotrophic influences to maintain higher τ values in older cultures. Since this requirement is not found in cultured fibers, τ would tend to assume slower values approaching those of P without hormone activation.