SARS-CoV-2 ORF8 as a modulator of cytokine induction : evidence and search for molecular mechanisms

Funding: This study was supported by Federal University of ABC (UFABC), São Bernardo do Campo, São Paulo, Brazil and by Wellcome Trust ISSF 204821/Z/16/Z to Michael Nevels (Universityof St Andrews, UK).Severe cases of SARS-CoV-2 infection are characterized by an immune response that leads to the overproduction of pro-inflammatory cytokines, resulting in lung damage, cardiovascular symptoms, hematologic symptoms, acute kidney injury and multiple organ failure that can lead to death. This remarkable increase in cytokines and other inflammatory molecules is primarily caused by viral proteins, and particular interest has been given to ORF8, a unique accessory protein specific to SARS-CoV-2. Despite plenty of research, the precise mechanisms by which ORF8 induces proinflammatory cytokines are not clear. Our investigations demonstrated that ORF8 augments production of IL-6 induced by Poly(I:C) in human embryonic kidney (HEK)-293 and monocyte-derived dendritic cells (mono-DCs). We discuss our findings and the multifaceted roles of ORF8 as a modulator of cytokine response, focusing on type I interferon and IL-6, a key component of the immune response to SARS-CoV-2. In addition, we explore the hypothesis that ORF8 may act through pattern recognition receptors of dsRNA such as TLRs.Publisher PDFPeer reviewe

In vitro susceptibility of Plasmodium falciparum Welch field isolates to infusions prepared from Artemisia annua L. cultivated in the Brazilian Amazon

Artemisinin is the active antimalarial compound obtained from the leaves of Artemisia annua L. Artemisinin, and its semi-synthetic derivatives, are the main drugs used to treat multi-drug-resistant Plasmodium falciparum (one of the human malaria parasite species). The in vitro susceptibility of P. falciparum K1 and 3d7 strains and field isolates from the state of Amazonas, Brazil, to A. annua infusions (5 g dry leaves in 1 L of boiling water) and the drug standards chloroquine, quinine and artemisinin were evaluated. The A. annua used was cultivated in three Amazon ecosystems (várzea, terra preta de índio and terra firme) and in the city of Paulínia, state of São Paulo, Brazil. Artemisinin levels in the A. annua leaves used were 0.90-1.13% (m/m). The concentration of artemisinin in the infusions was 40-46 mg/L. Field P. falciparum isolates were resistant to chloroquine and sensitive to quinine and artemisinin. The average 50% inhibition concentration values for A. annua infusions against field isolates were 0.11-0.14 μL/mL (these infusions exhibited artemisinin concentrations of 4.7-5.6 ng/mL) and were active in vitro against P. falciparum due to their artemisinin concentration. No synergistic effect was observed for artemisinin in the infusions

Genotyping of two Neisseria gonorrhoeae fluroquinolone-resistant strains in the Brazilian Amazon Region

We report two ciprofloxacin and ofloxacin-resistant Neisseria gonorrhoeae strains that were isolated from the urethral discharge of male patients at the sexually transmitted diseases outpatient clinic of the Alfredo da Matta Foundation (Manaus, state of Amazonas, Brazil). The gonococci displayed minimal inhibitory concentrations (> 32.00 µg/mL) and three mutations in the quinolone resistance-determining region (S91F and D95G in GyrA and S87R in ParC). Both isolates were genotyped using N. gonorrhoeae multi-antigen sequence typing and the analysis showed that the ST225 which represented an emerging widespread multi-resistant clone that has also been associated with reduced susceptibility to ceftriaxone. We recommend continued surveillance of this pathogen to assess the efficacy of anti-gonococcal antibiotics in Brazil

In vitro and in vivo antimalarial activity and cytotoxicity of extracts, fractions and a substance isolated from the amazonian plant tachia grandiflora (Gentianaceae)

Tachia sp. are used as antimalarials in the Amazon Region and in vivo antimalarial activity of a Tachia sp. has been previously reported. Tachia grandiflora Maguire and Weaver is an Amazonian antimalarial plant and herein its cytotoxicity and antimalarial activity were investigated. Spectral analysis of the tetraoxygenated xanthone decussatin and the iridoid aglyone amplexine isolated, respectively, from the chloroform fractions of root methanol and leaf ethanol extracts was performed. In vitro inhibition of the growth of Plasmodium falciparum Welch was evaluated using optical microscopy on blood smears. Crude extracts of leaves and roots were inactive in vitro. However, chloroform fractions of the root and leaf extracts [half-maximal inhibitory concentration (IC50) = 10.5 and 35.8 μg/mL, respectively] and amplexine (IC50 = 7.1 μg/mL) were active in vitro. Extracts and fractions were not toxic to type MRC-5 human fibroblasts (IC50 > 50 μg/mL). Water extracts of the roots of T. grandiflora administered by mouth were the most active extracts in the Peters 4-day suppression test in Plasmodium berghei-infected mice. At 500 mg/kg/day, these extracts exhibited 45-59% inhibition five to seven days after infection. T. grandiflora infusions, fractions and isolated substance have potential as antimalarials

Nursing diagnoses for the elderly using the International Classification for Nursing Practice and the activities of living model

OBJECTIVE: To develop nursing diagnosis statements for the elderly based on the Activities of Living Model and on the International Classification for Nursing Practice. METHOD: Descriptive and exploratory study, put in practice in two stages: 1) collection of terms and concepts that are considered clinically and culturally relevant for nursing care delivered to the elderly, in order to develop a database of terms and 2) development of nursing diagnosis statements for the elderly in primary health care, based on the guidelines of the International Council of Nurses and on the database of terms for nursing practice involving the elderly. RESULTS: 414 terms were identified and submitted to the content validation process, with the participation of ten nursing experts, which resulted in 263 validated terms. These terms were submitted to cross mapping with the terms of the International Classification for Nursing Practice, resulting in the identification of 115 listed terms and 148 non-listed terms, which constituted the database of terms, from which 127 nursing diagnosis statements were prepared and classified into factors that affect the performance of the elderly's activities of living - 69 into biological factors, 19 into psychological, 31 into sociocultural, five into environmental, and three into political-economic factors. CONCLUSIONS: After clinical validation, these statements can serve as a guide for nursing consultations with elderly patients, without ignoring clinical experience, critical thinking and decision-making

In vitro inhibition of Plasmodium falciparum by substances isolated from Amazonian antimalarial plants

In the present study, a quassinoid, neosergeolide, isolated from the roots and stems of Picrolemma sprucei (Simaroubaceae), the indole alkaloids ellipticine and aspidocarpine, isolated from the bark of Aspidosperma vargasii and A. desmanthum (Apocynaceae), respectively, and 4-nerolidylcatechol, isolated from the roots of Pothomorphe peltata (Piperaceae), all presented significant in vitro inhibition (more active than quinine and chloroquine) of the multi-drug resistant K1 strain of Plasmodium falciparum. Neosergeolide presented activity in the nanomolar range. This is the first report on the antimalarial activity of these known, natural compounds. This is also the first report on the isolation of aspidocarpine from A. desmanthum. These compounds are good candidates for pre-clinical tests as novel lead structures with the aim of finding new antimalarial prototypes and lend support to the traditional use of the plants from which these compounds are derived

In vitro susceptibility of Plasmodium falciparum Welch field isolates to infusions prepared from Artemisia annua L. cultivated in the Brazilian Amazon

Artemisinin is the active antimalarial compound obtained from the leaves of Artemisia annua L. Artemisinin, and its semi-synthetic derivatives, are the main drugs used to treat multi-drug-resistant Plasmodium falciparum (one of the human malaria parasite species). The in vitro susceptibility of P. falciparum K1 and 3d7 strains and field isolates from the state of Amazonas, Brazil, to A. annua infusions (5 g dry leaves in 1 L of boiling water) and the drug standards chloroquine, quinine and artemisinin were evaluated. The A. annua used was cultivated in three Amazon ecosystems (várzea, terra preta de índio and terra firme) and in the city of Paulínia, state of São Paulo, Brazil. Artemisinin levels in the A. annua leaves used were 0.90-1.13% (m/m). The concentration of artemisinin in the infusions was 40-46 mg/L. Field P. falciparum isolates were resistant to chloroquine and sensitive to quinine and artemisinin. The average 50% inhibition concentration values for A. annua infusions against field isolates were 0.11-0.14 μL/mL (these infusions exhibited artemisinin concentrations of 4.7-5.6 ng/mL) and were active in vitro against P. falciparum due to their artemisinin concentration. No synergistic effect was observed for artemisinin in the infusions

Mulheres vítimas de violência sexual: meios coercitivos e produção de lesões não-genitais Female victims of sexual abuse: coercive methods and non-genital injuries

Este estudo tem como objetivo analisar os meios coercitivos utilizados pelos ofensores sexuais e a produção de lesões não-genitais em mulheres crianças, adolescentes e adultas, vítimas de estupro e atentado violento ao pudor notificados pela Delegacia de Defesa da Mulher e examinados no Núcleo de Perícias Médico-Legais de Ribeirão Preto, São Paulo, Brasil, no período de 1996 a 2000. Na distribuição dos casos segundo a idade, utilizou-se o critério estabelecido pelo Estatuto da Criança e do Adolescente. A violência física foi o constrangimento mais empregado contra adultas (44,1%) e adolescentes (25,0%), seguida da grave ameaça em 36,5% e 17,0% dos casos, respectivamente. A violência presumida pela innocentia consilii ocorreu em 94,1% das crianças e 42,8% das adolescentes. Lesões não-genitais de natureza leve foram evidenciadas em 7,8% dos casos, acometendo crianças (3,0%), adolescentes (7,2%) e adultas (14,4%), produzidas sem o uso de armas (75,0%). Quando utilizadas, constatou-se que o número de casos com lesões decresce frente ao emprego de facas (14,3%) e revólveres (10,7%) contra vítimas adultas e adolescentes.This study aims to analyze the coercive methods used by sex offenders and the occurrence of non-genital injuries in female children, adolescent, and adult victims of sexual abuse reported to the Women's Police Precinct and examined at the Department of Forensic Medicine in Ribeirão Preto, São Paulo State, Brazil, from 1996 to 2000. Age distribution followed the criteria established by Brazil's Statute for Children and Adolescents (the prevailing legislation on issues pertaining to minors). Physical violence was the most frequently used form of coercion against adults (44.1%) and adolescents (25.0%), followed by serious threat in 36.5% and 17.0% of the cases, respectively. Presumed violence by innocentia consilii occurred in 94.1% of the children and 42.8% of the adolescents. Minor non-genital injuries were found in 7.8% of the cases, involving children (3.0%), adolescents (7.2%), and adults (14.4%), with no weapons involved in 75.0% of these cases. A decrease in the number of cases with injuries was observed in relation to knives (14.3%) and handguns (10.7%) used against adult and adolescent victims