Self-perception of voice and knowledge of vocal health and hygiene in Candomblé religious leaders in Brazil. / Autopercepção da voz e conhecimento em saúde e higiene vocal de líderes religiosos do Candomblé no Brasil.

PURPOSE: To verify possible complaints, voice and aerodigestive symptoms, singing voice handicap, and knowledge of vocal health and hygiene in Candomblé religious leaders in Brazil. METHODS: The study comprised 112 individuals who filled out a questionnaire with their identification and characterization, the stratified classification of their professional activities, and their self-perception of voice. Three self-assessment protocols - VoiSS, QSHV, and MSHI - were also used. RESULTS: The self-assessment of voice ranged from average to good. VoiSS mean total score was 23.04, which is above the cutoff. QSHV mean score was 23.54 points, which is near the cutoff. MSHI mean score (the perception of singing voice handicap) was 25.66 points. There was a substantially strong positive correlation between VoiSS and MSHI total scores (0.789; p<0.001). Women had higher limitation scores (p=0.012) and total scores (p=0.012) in VoiSS and higher handicap scores (p=0.038) in MSHI. Level I professionals - vocal elite (singers and actors) - had significantly higher QSHV scores than those in levels IV (p=0.010) and V (p=0.008). Most respondents had not visited an otorhinolaryngologist (89.29%) within the last year and had not been submitted to speech therapy (83.04%) for voice complaints. CONCLUSION: Candomblé leaders, particularly women, perceived voice symptoms and singing voice handicaps, with no relationship with their knowledge of vocal health and hygiene. Despite the complaints, most subjects reported not having visited health professionals responsible for voice care within the last year.

OBJETIVO: Verificar a autopercepção de sintomas vocais e aerodigestivos, desvantagem vocal no canto e o conhecimento em saúde e higiene vocal em líderes religiosos do Candomblé do Brasil. MÉTODO: Participaram deste estudo 112 indivíduos, que preencheram virtualmente um questionário de identificação, caracterização, classificação das atividades profissionais em níveis e autoavaliação vocal no momento atual, além de três protocolos de autoavaliação: Escala de Sinais e Sintomas (ESV), Questionário de Saúde e Higiene Vocal (QSHV) e Índice de Desvantagem para o Canto Moderno (IDCM). Foram utilizados os testes de correlação de Spearman, teste de Mann-Whitney e teste de Kruskal-Wallis. RESULTADOS: A autoavaliação vocal no momento atual variou de razoável a boa. A média do escore total na ESV foi de 23,04, acima da nota de corte. O QSHV apresentou valor médio de 23,54 pontos, próximo à nota de corte. O Índice de Desvantagem para o Canto Moderno (IDCM) mostrou média de 25,66 pontos. Houve correlação positiva de força substancial entre os escores totais da ESV e IDCM (0,789). Mulheres apresentaram maiores escores no ESV total e ESV limitação, além de maior escore no IDCM desvantagem. Profissionais do nível I - elite vocal (cantores e atores) apresentaram escores significativamente maiores para o QSHV que os profissionais do grupo IV (usuário não profissional não-vocal) e os do nível V (pessoas que estão fora do mercado de trabalho). A maioria dos respondentes não consultou Otorrinolaringologista (89,29%) no último ano e nem realizou fonoterapia (83,04%) por queixas vocais. CONCLUSÃO: Líderes do Candomblé apresentaram percepção de sintomas vocais e desvantagem vocal no canto, mais evidente nas mulheres, não havendo relação com o conhecimento de saúde e higiene vocal. Apesar da identificação de sintomas vocais, a maior parte dos sujeitos relatou não ter acessado profissionais de saúde responsáveis por esses cuidados com a voz no último ano.

Interleukin 17F Gene Polymorphism as a Potential Protective Factor in the Immunopathology of Ocular Toxoplasmosis.

Ocular toxoplasmosis (OT) is characterised by intraocular inflammation due to Toxoplasma gondii infection. Studies have found that interleukin 17 (IL-17) plays a central role in the pathology of OT. However, nucleotide variability in IL17 and interleukin 17 receptor (IL17R) genes has not been characterised in OT. As cytokine gene polymorphisms may influence the expression of these molecules, the aim of this study was to verify whether IL17A (rs2275913), IL17F (rs763780), IL17RA (rs4819554) and IL17RC (rs708567) polymorphisms are associated with OT in a Brazilian population. This study enrolled 214 patients seropositive for T. gondii (110 with OT and 104 without) and 107 controls. Polymorphisms were identified by PCR-restriction fragment length polymorphism analysis, validated by DNA sequencing with chi-square and multivariate analyses being used to assess possible associations between polymorphisms and OT. Logistic regression under the dominant model revealed a protection factor against OT of the C mutant allele of the IL17F (rs763780) polymorphism. The T/C-C/C genotypes were significantly more common in patients without OT compared to those with OT (p value = 0.0066) and controls (p value = 0.014). Findings from this study suggest that the IL17F polymorphism may have an influence in the immunopathology of OT in Brazilian individuals.

The impact of colonialism on head and neck cancer in Brazil: a historical essay focussing on tobacco, alcohol and slavery.

Colonialism's enduring impact on Brazil has had significant implications for health and oncology outcomes. This historical essay delves into the profound changes brought about by the transatlantic slave trade from Africa to the Americas, particularly in terms of its influence on the economy, sociocultural habits, and health outcomes. This essay explores the enduring connections between the colonial period's operational dynamics in Brazil and the current epidemiological panorama of head and neck cancer (HNC). The examination provides original insights on the role of tobacco and alcohol production and consumption, alongside the investigation of structural racism, which contributes to disparities in access to diagnosis, treatment, and prognosis for patients with HNC. This article presents novel visions and an analysis of evidence-based strategies to disrupt the adverse impact of colonialism's legacy on the epidemiology of HNC in Brazil.

Seroepidemiology of Toxoplasma gondii infection in blood donors in a population from the northwestern region of São Paulo state, Brazil.

BACKGROUND: Toxoplasmosis is one of the most common parasitic infections worldwide with varying prevalence between human populations. These variations are mainly associated with human exposure to risk factors. In this article, the seroprevalence of Toxoplasma gondii infection and the risk factors associated with infection in 1729 blood donors from São José do Rio Preto, São Paulo, Brazil were analysed. METHODS: The serological tests for detecting immunoglobulin M (IgM) and immunoglobulin G (IgG) anti-T. gondii were used. The risk factors associated with the infection were identified through the application of an epidemiological questionnaire. RESULTS: The prevalence of T. gondii infection was 48.0%. The following factors were identified in the final model after multiple logistic regression analysis: drinking raw milk (p=0.003; odds ratio [OR] 1.364 [confidence interval {CI} 1.1 to 1.7]), residing in a rural area (p<0.0001; OR 2.764 [CI 1.7 to 4.6]) and receiving a blood transfusion (p=0.015; OR 1.856 [CI 1.1 to 3.0]). CONCLUSIONS: The data obtained in this study showed that the blood donor population is exposed to risk factors related to infection by T. gondii. These data allow the establishment of control programs to contribute to public health in northwestern São Paulo state.

Apoptotic effect of Bulbine Natalensis and Chlorophytum Comosum in myelogenous Leukemia K562 cell line

Abstract Bulbine natalensis and Chorophytum comosum are potential medicinal source for the treatment of cancers. Chronic myeloid leukaemia is a hematopoietic stem cells disorder treated by tyrosine kinase inhibitors but often cause recurrence of the leukaemia after cessation of therapy, hence require alternative treatment. This study determines the anti-cancer effect of leaf, root and bulb methanolic and aqueous extracts of B. natalensis and C. comosum in chronic human myelogenous leukaemia (K562) cell line by MTT, Hoechst bis-benzimide nuclear and annexin V stain assays. The root methanolic extract of B. natalensis and C. comosum showed a high cytotoxicity of 8.6% and 16.7% respectively on the K562 cell line at 1,000 g/ml concentration. Morphological loss of cell membrane integrity causing degradation of the cell and fragmentation were observed in the root methanolic extract of both plants. A high apoptosis (p 0.0001) was induced in the K562 cells by both leaf and root extracts of the C. comosum compared to the B. natalensis. This study shows both plants possess apoptotic effect against in vitro myelogenous leukaemia which contributes to the overall anti-cancer properties of B. natalensis and C. comosum to justify future therapeutic applications against chronic myelogenous leukaemia blood cancer.

Resumo Bulbine natalensis Baker e Chorophytum comosum (Thunb.) Jacques são potenciais fontes medicinais para o tratamento de cânceres. A Leucemia Mieloide Crônica (LMC) é um distúrbio das células-tronco hematopoiéticas que é tratado com inibidores da tirosina quinase, mas frequentemente, causa recorrência da leucemia após a interrupção da terapia, portanto, requer um tratamento alternativo. Este estudo determinou o efeito anticancerígeno de extratos metanólicos e aquosos de folha, raiz e bulbo de B. natalensis e C. comosum na linhagem celular de leucemia mieloide humana crônica (K562) por ensaios de MTT, Hoechst bis-benzimida nuclear e anexina V. O extrato metanólico da raiz de B. natalensis e C. comosum apresentou alta citotoxidade de 8,6% e 16,7% respectivamente, na linhagem celular K562 com a concentração de 1,000 g / ml. Perda morfológica da integridade da membrana celular causando degradação dos núcleos, citoplasma e encolhimento celular foi observada no extrato metanólico da raiz de ambas as plantas. Uma alta apoptose (p 0,0001) foi induzida nas células K562 por extratos de folhas e raízes de C. comosum em comparação com B. natalensis. Este estudo mostrou que ambas as plantas possuem efeito apoptótico contra leucemia mieloide in vitro que contribui para as propriedades anticâncer gerais de B. natalensis e C. comosum para justificar futuras aplicações terapêuticas contra câncer de sangue de LMC.

Apoptotic effect of Bulbine Natalensis and Chlorophytum Comosum in myelogenous Leukemia K562 cell line / Efeito apoptótico da natalensia bulbina e do clorofito comoso na linhagem celular mielogênea da leucemia K562

Abstract Bulbine natalensis and Chorophytum comosum are potential medicinal source for the treatment of cancers. Chronic myeloid leukaemia is a hematopoietic stem cells disorder treated by tyrosine kinase inhibitors but often cause recurrence of the leukaemia after cessation of therapy, hence require alternative treatment. This study determines the anti-cancer effect of leaf, root and bulb methanolic and aqueous extracts of B. natalensis and C. comosum in chronic human myelogenous leukaemia (K562) cell line by MTT, Hoechst bis-benzimide nuclear and annexin V stain assays. The root methanolic extract of B. natalensis and C. comosum showed a high cytotoxicity of 8.6% and 16.7% respectively on the K562 cell line at 1,000 μg/ml concentration. Morphological loss of cell membrane integrity causing degradation of the cell and fragmentation were observed in the root methanolic extract of both plants. A high apoptosis (p < 0.0001) was induced in the K562 cells by both leaf and root extracts of the C. comosum compared to the B. natalensis. This study shows both plants possess apoptotic effect against in vitro myelogenous leukaemia which contributes to the overall anti-cancer properties of B. natalensis and C. comosum to justify future therapeutic applications against chronic myelogenous leukaemia blood cancer.

Resumo Bulbine natalensis Baker e Chorophytum comosum (Thunb.) Jacques são potenciais fontes medicinais para o tratamento de cânceres. A Leucemia Mieloide Crônica (LMC) é um distúrbio das células-tronco hematopoiéticas que é tratado com inibidores da tirosina quinase, mas frequentemente, causa recorrência da leucemia após a interrupção da terapia, portanto, requer um tratamento alternativo. Este estudo determinou o efeito anticancerígeno de extratos metanólicos e aquosos de folha, raiz e bulbo de B. natalensis e C. comosum na linhagem celular de leucemia mieloide humana crônica (K562) por ensaios de MTT, Hoechst bis-benzimida nuclear e anexina V. O extrato metanólico da raiz de B. natalensis e C. comosum apresentou alta citotoxidade de 8,6% e 16,7% respectivamente, na linhagem celular K562 com a concentração de 1,000 μg / ml. Perda morfológica da integridade da membrana celular causando degradação dos núcleos, citoplasma e encolhimento celular foi observada no extrato metanólico da raiz de ambas as plantas. Uma alta apoptose (p <0,0001) foi induzida nas células K562 por extratos de folhas e raízes de C. comosum em comparação com B. natalensis. Este estudo mostrou que ambas as plantas possuem efeito apoptótico contra leucemia mieloide in vitro que contribui para as propriedades anticâncer gerais de B. natalensis e C. comosum para justificar futuras aplicações terapêuticas contra câncer de sangue de LMC.

Establecimiento del esquema de purificación de los antígenos de SOBERANA®02, SOBERANA®Plus y SOBERANA 01 / Establishment of the purification scheme for SOBERANA®02, SOBERANA®Plus and SOBERANA 01 antigens

La urgente necesidad de desarrollar y producir vacunas seguras y efectivas para garantizar la reducción de la propagación del coronavirus de tipo 2 causante del síndrome respiratorio agudo severo, hizo que el Centro de Inmunología Molecular y el Instituto Finlay de Vacunas, desarrollaran dos vacunas y un candidato vacunal contra la COVID-19, que tienen como componente la molécula del dominio de unión al receptor (aa 319-541) del virus. Para establecer el proceso productivo, se realizaron experimentos en los posibles pasos del proceso de purificación de la molécula del dominio de unión al receptor (aa 319-541), con vistas a su posterior transferencia tecnológica a escala industrial. Dicha molécula está fusionada con una etiqueta de hexahistidina en su extremo C-terminal y presenta nueve residuos de cisteína en su secuencia que forman cuatro enlaces disulfuros intramoleculares, quedando una cisteína libre que permite obtener dos moléculas: dimérica y monomérica, antígenos que forman parte de las vacunas SOBERANA®02 y SOBERANA®Plus y el candidato vacunal SOBERANA 01. Se determinaron las mejores condiciones de adsorción de las matrices cromatográficas de afinidad por quelatos metálicos, intercambio catiónico y exclusión molecular. Se evaluó el desempeño del proceso a escala piloto y se caracterizó la molécula de acuerdo a sus propiedades físico-químicas y biológicas. Los resultados obtenidos mostraron un 60,02 ± 5,15por ciento de recuperación total de la proteína de interés, con más del 98% de pureza en ambas moléculas, una eficiente remoción de contaminantes y una antigenicidad mayor del 90por ciento referido al monómero control del dominio de unión al receptor con 99 por ciento de pureza, lo que demuestra que el proceso establecido es eficiente en la obtención de un producto con la calidad requerida(AU)

The urgent need to develop and produce safe and effective vaccines to guarantee the reduction of the spread of the type 2 coronavirus that causes severe acute respiratory syndrome, led the Center for Molecular Immunology and the Finlay Vaccine Institute to develop two vaccines and one candidate vaccine to combat the 2019 coronavirus pandemic. As part of the establishment of the production process, experiments were carried out on the possible steps of the purification process of the receptor binding domain molecule (aa 319-541) with a view to its subsequent technological transfer on an industrial scale. This molecule is fused with a hexahistidine tag at its C-terminal end and has nine cysteine residues in its sequence that form four intramolecular disulfide bonds; leaving a free cysteine that allows two molecules to be obtained: dimeric and monomeric, which constitute the antigens of the SOBERANA®02 and SOBERANA®Plus vaccines and the SOBERANA 01 vaccine candidate. The best adsorption conditions of the chromatographic matrices of affinity for metal chelates, cationic exchange and molecular exclusion were determined. The performance of the process was evaluated on a pilot scale and the molecule was characterized according to its physical-chemical and biological properties. The results obtained showed a 60.02 ± 5.15percent total recovery of the protein of interest with more than 98% purity in both molecules, an efficient removal of contaminants and an antigenicity greater than 90percent referred to the control monomer of the domain receptor binding with 99% purity; which demonstrates that the established process is efficient in obtaining a product with the required quality(AU)

Evaluation of the relationship between the STOP-Bang score with oxygen reserve index and difficult airway: a prospective observational study

Abstract Background: Patients diagnosed with Obstructive Sleep Apnea (OSA) syndrome have a tendency towards hypoventilation, hypoxia, and hypercarbia in the perioperative period. This study hypothesized that the Oxygen Reserve Index (ORi) could predict possible hypoxia and determine difficult airways in patients at risk for OSA, as determined by the STOP-Bang questionnaire. Methods: This prospective study included adult patients undergoing elective surgery under general anesthesia with endotracheal intubation, divided into two groups: low risk (0-2 points) and high risk (3-8 points) based on their STOP-Bang questionnaire results. The primary outcome measure was the highest ORi value reached during preoxygenation and the time to reach this value. Data were recorded at four time points: before preoxygenation (T1), end of preoxygenation (T2), end of mask ventilation (T3), and end of intubation (T4), as well as partial oxygen pressure values in T1, T2, and T4. The secondary outcome measures were the grading scale for mask ventilation, Cormack-Lehane score, tonsil dimensions, use of a stylet, and application of the burp maneuver during intubation. Results: In the high-risk group, preoperative peripheral oxygen saturation values, the highest ORi value reached in preoxygenation, and ORi values at T3 and T4 times were lower, and the time to reach the highest ORi value was longer (p < 0.05). Conclusion: Using ORi in patients with OSA may be useful in evaluating oxygenation, and since difficult airway is more common, ORi monitoring will better manage possible hypoxic conditions.

Development of a scalable single process for producing SARS-CoV-2 RBD monomer and dimer vaccine antigens.

We have developed a single process for producing two key COVID-19 vaccine antigens: SARS-CoV-2 receptor binding domain (RBD) monomer and dimer. These antigens are featured in various COVID-19 vaccine formats, including SOBERANA 01 and the licensed SOBERANA 02, and SOBERANA Plus. Our approach involves expressing RBD (319-541)-His6 in Chinese hamster ovary (CHO)-K1 cells, generating and characterizing oligoclones, and selecting the best RBD-producing clones. Critical parameters such as copper supplementation in the culture medium and cell viability influenced the yield of RBD dimer. The purification of RBD involved standard immobilized metal ion affinity chromatography (IMAC), ion exchange chromatography, and size exclusion chromatography. Our findings suggest that copper can improve IMAC performance. Efficient RBD production was achieved using small-scale bioreactor cell culture (2 L). The two RBD forms - monomeric and dimeric RBD - were also produced on a large scale (500 L). This study represents the first large-scale application of perfusion culture for the production of RBD antigens. We conducted a thorough analysis of the purified RBD antigens, which encompassed primary structure, protein integrity, N-glycosylation, size, purity, secondary and tertiary structures, isoform composition, hydrophobicity, and long-term stability. Additionally, we investigated RBD-ACE2 interactions, in vitro ACE2 recognition of RBD, and the immunogenicity of RBD antigens in mice. We have determined that both the monomeric and dimeric RBD antigens possess the necessary quality attributes for vaccine production. By enabling the customizable production of both RBD forms, this unified manufacturing process provides the required flexibility to adapt rapidly to the ever-changing demands of emerging SARS-CoV-2 variants and different COVID-19 vaccine platforms.

Conformational characterization of the mammalian-expressed SARS-CoV-2 recombinant receptor binding domain, a COVID-19 vaccine.

The COVID-19 pandemic has caused a large number of diseases worldwide. There are few vaccines to constrain this disease and the value of them is high. In this sense, the antigens of the vaccine platform Soberana, the receptor binding domain from SARS-CoV-2 Spike protein, both the monomeric (mRBD) and dimeric (dRBD) forms, have been developed. This study encompassed several analyses by different techniques like circular dichroism (CD), fluorescence spectroscopy (FS) and Gel Filtration- High Performance Liquid ChLC of mRBD and dRBD. Monomer and dimer exhibited similar far-UV CD spectral characteristics with 54% of ß-sheet content. Similar conformational features according to near-UV CD and FS studies were observed in both RBD. Stress stability studies by far-UV CD, FS, biological activity and GF-HPLC at 37 °C showed that mRBD is very stable. On the other hand, dRBD fluorescent emission showed a shift towards higher wavelengths as the incubation time increases, suggesting exposition of tryptophan residues, unlike what happens with mRBD. Biological activity outcome confirms these results. GF-HPLC profiles showed that in mRBD, the product of molecular stress are dimers and does not increase over time. However, dRBD showed dimer fragmentation as the main degradation species. This study reveals the usefulness of CD techniques for the analysis of degradation of RBD molecules as well as showed the difference in stability of both RBD molecules. Besides, our work provides useful insights into the production of a key protein used in diagnosis and therapeutics to fight COVID-19 pandemia.

Role of interleukin 1ß and interleukin 10 variants on ocular toxoplasmosis in Brazilian individuals.

Introduction: Ocular toxoplasmosis (OT) is an intraocular inflammation caused by Toxoplasma gondii infection that affects the retina and choroid, giving rise to posterior uveitis. Genetic polymorphisms in cytokine genes may exert influence in the expression of these molecules and play a significant role in inflammatory responses and susceptibility to OT. The aim of this study was to evaluate the role of polymorphisms rs16944 (-511 C > T) of the interleukin (IL) 1ß gene and rs1800896 (-1082 G > A) of the IL10 gene on OT in Brazilian individuals with a serologic diagnosis of T. gondii and after conducting fundoscopic exams. Methods: Participants with a positive serology were classified into two distinct groups according to the presence (G1; n = 110) or absence (G2; n = 104) of OT. The control group (G3) consisted of individuals without the infection (n = 108). Results: It was observed that the C/C genotype of the IL1ß gene polymorphism was a protective factor for OT (p = 0.02, OR = 0.28, 95% CI 0.08-0.78 for G1 vs. G2; p = 0.03; OR = 0.29, 95% CI 0.09-0.82 for G1 vs. G3), according to the recessive inheritance model. Conclusions: The -511C.T polymorphisms of the IL1ß gene seems to play an important role in the pathogenesis of OT in Brazilian individuals.

Conformational characterization of the mammalian-expressed SARS-CoV-2 recombinant receptor binding domain, a COVID-19 vaccine

The COVID-19 pandemic has caused a large number of diseases worldwide. There are few vaccines to constrain this disease and the value of them is high. In this sense, the antigens of the vaccine platform Soberana, the receptor binding domain from SARS-CoV-2 Spike protein, both the monomeric (mRBD) and dimeric (dRBD) forms, have been developed. This study encompassed several analyses by different techniques like circular dichroism (CD), fluorescence spectroscopy (FS) and Gel Filtration- High Performance Liquid ChLC of mRBD and dRBD. Monomer and dimer exhibited similar far-UV CD spectral characteristics with 54% of ß-sheet content. Similar conformational features according to near-UV CD and FS studies were observed in both RBD. Stress stability studies by far-UV CD, FS, biological activity and GF-HPLC at 37 °C showed that mRBD is very stable. On the other hand, dRBD fluorescent emission showed a shift towards higher wavelengths as the incubation time increases, suggesting exposition of tryptophan residues, unlike what happens with mRBD. Biological activity outcome confirms these results. GF-HPLC profiles showed that in mRBD, the product of molecular stress are dimers and does not increase over time. However, dRBD showed dimer fragmentation as the main degradation species. This study reveals the usefulness of CD techniques for the analysis of degradation of RBD molecules as well as showed the difference in stability of both RBD molecules. Besides, our work provides useful insights into the production of a key protein used in diagnosis and therapeutics to fight COVID-19 pandemia.

Safety and immunogenicity of anti-SARS-CoV-2 heterologous scheme with SOBERANA 02 and SOBERANA Plus vaccines: Phase IIb clinical trial in adults.

BACKGROUND: SOBERANA 02 has been evaluated in phase I and IIa studies comparing homologous versus heterologous schedule (this one, including SOBERANA Plus). Here, we report results of immunogenicity, safety, and reactogenicity of SOBERANA 02 in a two- or three-dose heterologous scheme in adults. METHOD: Phase IIb was a parallel, multicenter, adaptive, double-blind, randomized, and placebo-controlled trial. Subjects (n = 810) aged 19-80 years were randomized to receive two doses of SARS-CoV-2 RBD conjugated to tetanus toxoid (SOBERANA 02) and a third dose of dimeric RBD (SOBERANA Plus) 28 days apart; two production batches of active ingredients of SOBERANA 02 were evaluated. Primary outcome was the percentage of seroconverted subjects with ≥4-fold the anti-RBD immunoglobulin G (IgG) concentration. Secondary outcomes were safety, reactogenicity, and neutralizing antibodies. FINDINGS: Seroconversion rate in vaccinees was 76.3% after two doses and 96.8% after the third dose of SOBERANA Plus (7.3% in the placebo group). Neutralizing IgG antibodies were detected against D614G and variants of concern (VOCs) Alpha, Beta, Delta, and Omicron. Specific, functional antibodies were detected 7-8 months after the third dose. The frequency of serious adverse events (AEs) associated with vaccination was very low (0.1%). Local pain was the most frequent AE. CONCLUSIONS: Two doses of SOBERANA 02 were safe and immunogenic in adults. The heterologous combination with SOBERANA Plus increased neutralizing antibodies, detectable 7-8 months after the third dose. TRIAL REGISTRY: https://rpcec.sld.cu/trials/RPCEC00000347 FUNDING: This work was supported by Finlay Vaccine Institute, BioCubaFarma, and the Fondo Nacional de Ciencia y Técnica (FONCI-CITMA-Cuba, contract 2020-20).

Lack of association of the KIR and HLA class I ligands with ZIKV infection in south and southeast of Brazil.

BACKGROUND: Zika virus (ZIKV) is an emerging arbovirus associated with foetal malformations and neurological complications. The infection is usually associated with mild symptoms. The comparison between the allelic frequency of polymorphic genes in symptomatic infected individuals in the population can clarify the pathogenic mechanisms of ZIKV. During ZIKV infection, cytokines are produced and natural killer (NK) cells are recruited, whose activation depends on signaling pathways activated by specific receptors, such as killer cell immunoglobulin-like receptors (KIR). These molecules interact with human leukocyte antigen (HLA) class I ligands and are encoded by polymorphic genes. OBJECTIVES: This study aimed to evaluate the frequency of allelic variants of the genes encoding the KIR receptors and their HLA class I ligands in 139 symptomatic ZIKV-patients and 170 controls negative for the virus, and to evaluate the role of these variants for ZIKV susceptibility. METHODS: KIR and HLA class I genes were genotyped using the polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) technique. FINDINGS: No significant differences in the frequency distribution of KIRs and KIR-HLA in patients compared to controls were observed. MAIN CONCLUSIONS: KIR and its HLA ligands might play a minor role in ZIKV infection in the south and southeast Brazilian individuals.

Production of an anti-TNFα antibody in murine myeloma cells by perfusion culture.

Infliximab is a mouse/human chimeric IgG1 monoclonal antibody which recognizes the proinflammatory cytokine, tumor necrosis factor α (TNFα), and inhibits receptor interactions, thereby decreasing inflammation and autoimmune response in patients. This monoclonal antibody has been successfully used to treat rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis. However, the high treatment cost limits patient access to this biotherapy. One alternative to this problem is the use of biosimilars. In this work, we describe the stable expression and physicochemical characterization of an anti-TNFα antibody. While infliximab is produced in recombinant murine SP2/0 cells, our anti-TNFα IgG antibody was expressed in recombinant murine NS0 myeloma cells. The best anti-TNFα antibody-expressing clone was selected from three clone candidates based on the stability of IgG expression levels, specific productivity as well as TNFα-binding activity compared to commercial infliximab. Our results indicate that the selected cell clone, culture medium, and fermentation mode allowed for the production of an anti-TNFα antibody with similar characteristics to the reference commercially available product. An optimization of the selected culture medium by metabolomics may increase the volumetric productivity of the process to satisfy the demand for this product. Further experiments should be performed to evaluate the biological properties of this anti-TNFα antibody. KEY POINTS: • An anti-TNFα antibody was produced in NS0 cells using perfusion culture. • A proprietary chemically defined culture medium was used to replace commercially available protein-free medium. • The purified anti-TNFα antibody was comparable to the reference marketed product.

HLA-G, LILRB1 and LILRB2 Variants in Zika Virus Transmission from Mother to Child in a Population from South and Southeast of Brazil.

During the 2015-2016 epidemic, Brazil was the country with the highest rate of Zika virus (ZIKV) infection in the Americas. Twenty-nine percent of pregnant women positive for ZIKV exhibited ultrasound scans with fetus anomalies. Human leukocyte antigen-G (HLA-G) exerts immunoregulatory effects by binding to inhibitory receptors, namely LILRB1 and LILRB2, thus preventing mother-fetus rejection and vertical pathogen transmission. The binding of HLA-G to one of its receptors modulates both innate and adaptive immunity. However, in a viral infection, these molecules may behave as pathogenic mediators shifting the pregnancy environment from an anti-inflammatory profile to a pro-inflammatory phenotype. Genetic mutations might be associated with the change in phenotype. This study aimed to explore the possible role of polymorphic sites in HLA-G, LILRB1 and LILRB2 in mother-fetus ZIKV transmission. Polymorphisms were detected by direct sequencing. Differences in allele and/or genotype frequencies for each SNP analyzed among ZIKV non-transmitting and transmitting mother-child pairs, among ZIKV-transmitting and non-transmitting mothers and between ZIKV-infected and non-infected children were compared by Mid-P exact test or Yates' correction. Significant susceptibility of ZIKV vertical transmission is suggested in ZIKV-transmitting and non-transmitting mothers and ZIKV-infected and non-infected children for LILRB1_rs1061684 T/T (p = 0.03, Pc = 0.06, OR = 12.4; p = 0.008, Pc = 0.016, OR = 16.4) and LILRB1_rs16985478 A/A (p = 0.01, Pc = 0.02, OR = 19.2; p = 0.008, Pc = 0.016, OR = 16.4). HLA-G_rs1710 (p = 0.04, Pc = 0.52, OR = 4.30) was also a susceptibility factor. LILRB2_rs386056 G/A (p = 0.02, Pc = 0.08, OR = 0.07), LILRB2_rs7247451 G/G (p = 0.01, Pc = 0.04, OR = 0.04) and HLAG_rs9380142 T/T (p = 0.04, Pc = 0.52, OR = 0.14) were suggested as protective factors against vertical transmission. The current study suggests that polymorphic sites in the LILRB1 and HLA-G genes might be associated with mother-to-child ZIKV transmission while LILRB2 might be associated with protection against ZIKV transmission in the womb in a population from the south and southeast of Brazil.

Determination of the ursolic and oleanolic acids content with the antioxidant capacity in apple peel extract of various cultivars.

Apples are rich sources of ursolic acid (UA) and oleanolic acid (OA) which are the major and most prominent triterpenes in the peel of an apple. Pentacyclic triterpenes are ideal nutraceuticals due to their ability to reduce the risk of many life-threatening diseases such as cancer, cardiovascular and diabetes. This study was to determine the content of UA and OA in the apple peel extract from different cultivars grown in South Africa as well as the correlation of their content level with antioxidant capacity. Quantitative analysis of UA and OA in apple peels from three cultivars; red delicious (RD), royal gala (RG) and granny smith (GS) apples was carried out using HPLC and their antioxidant capacity was analyzed using the DPPH assay. The RD showed the highest content of UA and OA (248.02 ± 0.08 µg/ml and 110.00 ± 0.08 µg/ml respectively) in the apple peel extract and also displayed a significantly high level of antioxidant capacity (97.3 ± 0.40%; p < 0.0001) compared to the RG and GS cultivars. A strong positive correlation was noted between the UA, OA and antioxidant capacities of all the cultivars. Only the RD cultivar showed a significant correlation though; UA (r = 0.9570; p = 0.0027) and OA (r = 0.8503; p = 0.0319). This study demonstrated that the RD and RG apple peels possess the highest UA and OA content which invariably increases their antioxidant activities compared to GS apple. Thus, both apple cultivars would be useful and recommended for food consumption and nutraceuticals values to improve human health.

Safety and immunogenicity of anti-SARS CoV-2 vaccine SOBERANA 02 in homologous or heterologous scheme: Open label phase I and phase IIa clinical trials.

BACKGROUND: SOBERANA 02 is a COVID-19 vaccine based on SARS-CoV-2 recombinant RBD conjugated to tetanus toxoid (TT). SOBERANA Plus antigen is dimeric-RBD. Here we report safety and immunogenicity from phase I and IIa clinical trials using two-doses of SOBERANA 02 and three-doses (homologous) or heterologous (with SOBERANA Plus) protocols. METHOD: We performed an open-label, sequential and adaptive phase I to evaluate safety and explore the immunogenicity of SOBERANA 02 in two formulations (15 or 25 µg RBD-conjugated to 20 µg of TT) in 40 subjects, 19-59-years-old. Phase IIa was open-label including 100 volunteers 19-80-years, receiving two doses of SOBERANA 02-25 µg. In both trials, half of volunteers were selected to receive a third dose of the corresponding SOBERANA 02 and half received a heterologous dose of SOBERANA Plus. Primary outcome was safety. The secondary outcome was immunogenicity evaluated by anti-RBD IgG ELISA, molecular neutralization of RBD:hACE2 interaction, live-virus-neutralization and specific T-cells response. RESULTS: The most frequent adverse event (AE) was local pain, other AEs had frequencies ≤ 5%. No serious related-AEs were reported. Phase IIa confirmed the safety in 60 to 80-years-old subjects. In phase-I SOBERANA 02-25 µg elicited higher immune response than SOBERANA 02-15 µg and progressed to phase IIa. Phase IIa results confirmed the immunogenicity of SOBERANA 02-25 µg even in 60-80-years. Two doses of SOBERANA02-25 µg elicited an immune response similar to that of the Cuban Convalescent Serum Panel and it was higher after the homologous and heterologous third doses. The heterologous scheme showed a higher immunological response. Anti-RBD IgG neutralized the delta variant in molecular assay, with a 2.5-fold reduction compared to D614G neutralization. CONCLUSIONS: SOBERANA 02 was safe and immunogenic in persons aged 19-80 years, eliciting neutralizing antibodies and specific T-cell response. Highest immune responses were obtained in the heterologous three doses protocol. TRIAL REGISTRY: https://rpcec.sld.cu/trials/RPCEC00000340, https://rpcec.sld.cu/trials/RPCEC00000347.

TNFα rs1799964 TT genotype may be a susceptibility factor for vertical transmission of Toxoplasma gondii and clinical signs in newborns from pregnant women with acute toxoplasmosis.

BACKGROUND: One of the main impacts of Toxoplasma gondii infection occurs during pregnancy and is related to the vertical transmission of the parasite (congenital toxoplasmosis), which can cause severe clinical outcomes and fetal death. During acute infection, in order to control the rapid replication of tachyzoites, different host immune response genes are activated, and these include cytokine-encoding genes. Considering that polymorphisms in cytokine genes may increase susceptibility to vertical transmission of T. gondii by determining the immune status of the pregnant woman, this study evaluated the influence of polymorphisms of tumor necrosis factor alpha (TNFα) rs1799964 (- 1031) and interleukin 1 beta (IL1ß) rs16944 (- 511) genes on gestational toxoplasmosis and on the vertical transmission of the parasite and verified the allele and genotype frequency of these polymorphisms in pregnant patients whose respective newborn did or did not present clinical abnormalities suggestive of congenital toxoplasmosis. METHODS AND RESULTS: A total of 204 pregnant patients with (n = 114) or without (n = 90) infection by T. gondii were enrolled. No associations were found involving the polymorphisms rs1799964 (- 1031) of the TNFα gene and rs16944 (- 511) of the IL1ß gene with the increased chance of T. gondii infection during pregnancy. However, it was observed that the maternal TT genotype referring to the polymorphism of the TNFα gene seems to influence the vertical transmission of the parasite (P = 0.01; χ2 = 6.05) and the presence of clinical manifestation in newborns from pregnancies with acute toxoplasmosis (P = 0.007; χ2 = 9.68). CONCLUSION: The TNFα rs1799964 TT genotype may act as a susceptibility factor for the vertical transmission of parasite and for the presence of clinical signs in newborns from pregnant women with acute toxoplasmosis.