Lisinopril increases lung ACE2 levels and SARS-CoV-2 viral load and decreases inflammation but not disease severity in experimental COVID-19.

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.

Virulence Profiles of Wild-Type, P.1 and Delta SARS-CoV-2 Variants in K18-hACE2 Transgenic Mice.

Since December 2019, the world has been experiencing the COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and we now face the emergence of several variants. We aimed to assess the differences between the wild-type (Wt) (Wuhan) strain and the P.1 (Gamma) and Delta variants using infected K18-hACE2 mice. The clinical manifestations, behavior, virus load, pulmonary capacity, and histopathological alterations were analyzed. The P.1-infected mice showed weight loss and more severe clinical manifestations of COVID-19 than the Wt and Delta-infected mice. The respiratory capacity was reduced in the P.1-infected mice compared to the other groups. Pulmonary histological findings demonstrated that a more aggressive disease was generated by the P.1 and Delta variants compared to the Wt strain of the virus. The quantification of the SARS-CoV-2 viral copies varied greatly among the infected mice although it was higher in P.1-infected mice on the day of death. Our data revealed that K18-hACE2 mice infected with the P.1 variant develop a more severe infectious disease than those infected with the other variants, despite the significant heterogeneity among the mice.

Virulence profiles of wild-type, P.1 and delta SARS-CoV-2 variants in K18-hACE2 transgenic mice

Since December 2019, the world has been experiencing the COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and we now face the emergence of several variants. We aimed to assess the differences between the wild-type (Wt) (Wuhan) strain and the P.1 (Gamma) and Delta variants using infected K18-hACE2 mice. The clinical manifestations, behavior, virus load, pulmonary capacity, and histopathological alterations were analyzed. The P.1-infected mice showed weight loss and more severe clinical manifestations of COVID-19 than the Wt and Delta-infected mice. The respiratory capacity was reduced in the P.1-infected mice compared to the other groups. Pulmonary histological findings demonstrated that a more aggressive disease was generated by the P.1 and Delta variants compared to the Wt strain of the virus. The quantification of the SARS-CoV-2 viral copies varied greatly among the infected mice although it was higher in P.1-infected mice on the day of death. Our data revealed that K18-hACE2 mice infected with the P.1 variant develop a more severe infectious disease than those infected with the other variants, despite the significant heterogeneity among the mice.

A rabies virus vampire bat variant shows increased neuroinvasiveness in mice when compared to a carnivore variant

Rabies is one of the most important zoonotic diseases and is caused by several rabies virus (RABV) variants. These variants can exhibit differences in neurovirulence, and few studies have attempted to evaluate the neuroinvasiveness of variants derived from vampire bats and wild carnivores. The aim of this study was to evaluate the neuropathogenesis of infection with two Brazilian RABV street variants (variant 3 and crab-eating fox) in mice. BALB/c mice were inoculated with RABV through the footpad, with the 50% mouse lethal dose (LD50) determined by intracranial inoculation. The morbidity of rabies in mice infected with variant 3 and the crab-eating fox strain was 100% and 50%, respectively, with an incubation period of 7 and 6 days post-inoculation (dpi), respectively. The clinical disease in mice was similar with both strains, and it was characterized initially by weight loss, ruffled fur, hunched posture, and hind limb paralysis progressing to quadriplegia and recumbency at 9 to 12 dpi. Histological lesions within the central nervous system (CNS) characterized by nonsuppurative encephalomyelitis with neuronal degeneration and necrosis were observed in mice infected with variant 3 and those infected with the crab-eating fox variant. However, lesions and the presence of RABV antigen, were more widespread within the CNS of variant-3-infected mice, whereas in crab-eating fox-variant-infected mice, RABV antigens were more restricted to caudal areas of the CNS, such as the spinal cord and brainstem. In conclusion, the results shown here demonstrate that the RABV vampire bat strain (variant 3) has a higher potential for neuroinvasiveness than the carnivore variant. (AU) i

A rabies virus vampire bat variant shows increased neuroinvasiveness in mice when compared to a carnivore variant.

Rabies is one of the most important zoonotic diseases and is caused by several rabies virus (RABV) variants. These variants can exhibit differences in neurovirulence, and few studies have attempted to evaluate the neuroinvasiveness of variants derived from vampire bats and wild carnivores. The aim of this study was to evaluate the neuropathogenesis of infection with two Brazilian RABV street variants (variant 3 and crab-eating fox) in mice. BALB/c mice were inoculated with RABV through the footpad, with the 50% mouse lethal dose (LD50) determined by intracranial inoculation. The morbidity of rabies in mice infected with variant 3 and the crab-eating fox strain was 100% and 50%, respectively, with an incubation period of 7 and 6 days post-inoculation (dpi), respectively. The clinical disease in mice was similar with both strains, and it was characterized initially by weight loss, ruffled fur, hunched posture, and hind limb paralysis progressing to quadriplegia and recumbency at 9 to 12 dpi. Histological lesions within the central nervous system (CNS) characterized by nonsuppurative encephalomyelitis with neuronal degeneration and necrosis were observed in mice infected with variant 3 and those infected with the crab-eating fox variant. However, lesions and the presence of RABV antigen, were more widespread within the CNS of variant-3-infected mice, whereas in crab-eating fox-variant-infected mice, RABV antigens were more restricted to caudal areas of the CNS, such as the spinal cord and brainstem. In conclusion, the results shown here demonstrate that the RABV vampire bat strain (variant 3) has a higher potential for neuroinvasiveness than the carnivore variant.

Análise da textura de músculos submetidos à fixação em formaldeído e conservação em solução de benzoato de sódio 0, 5% e ácido acético 0, 5% / Tenderness analysis of muscles fixed in formaldehyde and kept in 0, 5% sodium benzoate and 0, 5% acetic acid solution

Background: Tissue texture, or tenderness, may be checked-out by analyzing the force necessary to cause muscle fibers rupture (shearing force) and the result is expressed in kilogram-force (kgf). The aim of this paper was to analyze the texture of broiler chicken breast muscles fixed in a 10% formaldehyde solution and kept in a 0.5% sodium benzoate and 0.5% acetic acid solution up to 180 days so any alteration regarding resistance and the best moment for dissection could be described. Materials, Methods & Results: Forty chicken breasts were used and muscle texture was analyzed by the shearing force whose values were correspondent to the maximum force necessary to chisel the samples. Eight of 40 breasts were taken as a control group (fresh muscles). The remaining breast muscles were fixed in a 4% formaldehyde solution and kept in plastic containers with this solution during seven days and, then, eight breasts went through texture analysis. The 24 remaining breasts were set in plastic containers with 0.5% sodium benzoate and 0.5% acetic acid solution. The texture analyses were taken out at 30, 90 and 180 days of conservation, and eight breasts were used in each time. The shearing force was obtained for each sample and, just before cutting, muscles were washed in running water for 24 h so the excess of the conservation agent could be removed. Data were submitted to statistical analysis by the 5% Tukey's test. The average shearing force of the control, formaldehyde and 30-day sodium benzoate and acetic acid groups differed among them and from the 90-day and 180-day of sodium benzoate and acetic acid groups. The average shearing forces of the two last groups did not statistically differ among them. Discussion: The longer the muscles were kept in the sodium benzoate and acetic acid solutions, the more they became soft, and the shearing force decreased from 21.46 Kgf (seven days of conservation) to 19.20 Kgf (30 days), 17.23 Kgf (90 days) and 16.32 Kgf (180 days) not reaching, however, values from 8.97 to 15.31 Kgf, as described in chicken breasts conserved in 96oGL ethylic alcohol. The average shearing force of chicken breasts conserved in sodium benzoate and acetic acid progressively decreased during 180 days of conservation, when it reached a value of 16.32 Kgf, similar to the value of 15.32 Kgf of chicken breasts conserved for 45 days in formaldehyde. By the seventh day of conservation in 10% formaldehyde, samples presented an average shearing force of 21.46 Kgf, which was higher than the 15.33 Kgf of chicken breasts conserved during 45 days or the 14.74 to 17.37 Kgf of the same tissue kept up to a year in formaldehyde solution. It was concluded that conservation of breast broiler chickens fixed in formaldehyde and conserved in sodium benzoate and acetic acid made them softer through within 180 days, resulting in a better structure malleability and suggesting that dissection of anatomical muscle pieces may occur in an easier way in this moment. Although the sodium benzoate and acetic acid effect in texture of broiler chicken muscle tissue fixed in formaldehyde has been studied, it is believed that, due this great tissue homogeneity, these data might be taken or used as basis to similar studies in other species.

Análise da textura de músculos peitorais submetidos à fixação e conservação em álcool / Tenderness analysis of muscles fixed and kept in alcohol

A textura, ou maciez, pode ser avaliada pela mensuração da força necessária para ocorrer o cisalhamento das fibras musculares. Objetivou-se, nesse trabalho, a análise da textura de músculos submetidos à fixação e conservação em álcool, ao longo de um ano, mediante uso de um aparelho analisador de textura. Foram utilizados 48 peitos de frangos jovens, pesados, fixados e conservados em álcool etílico 96° GL. As análises foram realizadas após 15, 30, 90, 180 e 360 dias de conservação, além do grupo controle de músculos frescos. Os valores da força de cisalhamento dos diferentes grupos aumentaram progressivamente de 3,38 (grupo controle) até 15,31 Kgf (180 dias), caindo para 9,53 Kgf após 360 dias. Concluiu-se que quando músculos são submetidos à fixação e conservação em álcool 96° GL, ocorre diminuição da maciez, tornando-os quase cinco vezes mais rígidos ao corte após seis meses, e três vezes mais rígidos após um ano. Sugere-se que a dissecção de peças anatômicas musculares ocorra até 90 dias após fixação e conservação em álcool 96° GL ou ao redor de um ano nesse agente conservante, pois há menor rigidez tissular nesses períodos. Embora se tenha estudado o efeito do álcool na textura de tecido muscular de aves, acredita-se que, devido à grande homogeneidade tissular neste caso, tais dados possam ser extrapolados ou servir de base para estudos similares em outras espécies.

Texture, or tenderness, may be evaluated by measuring the force necessary to cause rupture of muscle fibers. The aim of this paper was to analyze the texture of muscles fixed and kept in alcohol, throughout a year, by using a texture analyzer. Forty eight poultry breasts were weighted, fixed and kept in a 96° GL ethylic alcohol solution. Analyses were performed at 15, 30, 90, 180 and 360 days of conservation, besides the one of the control group of fresh breasts. Values of shear forces of different groups increased progressively from 3.38 (control group) to 15.31 Kgf (180 days), decreasing to 9.53Kgf after 360 days. It was concluded that when muscles are fixed and kept in ethylic alcohol 96° GL the tenderness is decreased, becoming almost five times harder during the first six months and three times harder after a year. It is suggested that muscle anatomic pieces dissection occurs up to 90 days after fixation and conservation in 96° GL alcohol or around 1 year on this conservation product because there is smaller muscle stiffness in these times. Although the alcohol effect in texture of poultry muscle tissue has being studied in this paper, it is believed that, due this great tissue homogeneity, these data might be taken or being basis to similar studies in other species.

Análise da textura de músculos peitorais submetidos à fixação e conservação em álcool / Tenderness analysis of muscles fixed and kept in alcohol

A textura, ou maciez, pode ser avaliada pela mensuração da força necessária para ocorrer o cisalhamento das fibras musculares. Objetivou-se, nesse trabalho, a análise da textura de músculos submetidos à fixação e conservação em álcool, ao longo de um ano, mediante uso de um aparelho analisador de textura. Foram utilizados 48 peitos de frangos jovens, pesados, fixados e conservados em álcool etílico 96° GL. As análises foram realizadas após 15, 30, 90, 180 e 360 dias de conservação, além do grupo controle de músculos frescos. Os valores da força de cisalhamento dos diferentes grupos aumentaram progressivamente de 3,38 (grupo controle) até 15,31 Kgf (180 dias), caindo para 9,53 Kgf após 360 dias. Concluiu-se que quando músculos são submetidos à fixação e conservação em álcool 96° GL, ocorre diminuição da maciez, tornando-os quase cinco vezes mais rígidos ao corte após seis meses, e três vezes mais rígidos após um ano. Sugere-se que a dissecção de peças anatômicas musculares ocorra até 90 dias após fixação e conservação em álcool 96° GL ou ao redor de um ano nesse agente conservante, pois há menor rigidez tissular nesses períodos. Embora se tenha estudado o efeito do álcool na textura de tecido muscular de aves, acredita-se que, devido à grande homogeneidade tissular neste caso, tais dados possam ser extrapolados ou servir de base para estudos similares em outras espécies.(AU)

Texture, or tenderness, may be evaluated by measuring the force necessary to cause rupture of muscle fibers. The aim of this paper was to analyze the texture of muscles fixed and kept in alcohol, throughout a year, by using a texture analyzer. Forty eight poultry breasts were weighted, fixed and kept in a 96° GL ethylic alcohol solution. Analyses were performed at 15, 30, 90, 180 and 360 days of conservation, besides the one of the control group of fresh breasts. Values of shear forces of different groups increased progressively from 3.38 (control group) to 15.31 Kgf (180 days), decreasing to 9.53Kgf after 360 days. It was concluded that when muscles are fixed and kept in ethylic alcohol 96° GL the tenderness is decreased, becoming almost five times harder during the first six months and three times harder after a year. It is suggested that muscle anatomic pieces dissection occurs up to 90 days after fixation and conservation in 96° GL alcohol or around 1 year on this conservation product because there is smaller muscle stiffness in these times. Although the alcohol effect in texture of poultry muscle tissue has being studied in this paper, it is believed that, due this great tissue homogeneity, these data might be taken or being basis to similar studies in other species.(AU)