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ABSTRACT Introduction: The identification of the wide tropism of SARS-CoV-2 can be critical for viral pathogenesis, and it is therefore relevant to determine the infection mechanisms and identify new therapeutic targets. This study consists of two experimental blocks: a) isolation and characterization of the SARS-CoV-2 virus, and b) pathogenesis, tropism, and targets, both molecular and therapeutic. Methods: The procedures used were the following: a) clinical samples were obtained from the nasopharyngeal cavity of COVID-19 positive patients, which were inoculated in Vero E6 cells and analyzed by Scanning Electron Microscopy 72 h after infection. b) They were obtained visceral organ samples (lung, liver, kidney and brain) from patients dyed from COVID-19. Cryocuts were prepared and evaluated by Masson's Trichrome Staining, Electron Microscopy and Confocal Microscopy. Results: Results show a) Extensive damage in infected Vero E6 cells where virus particles and extracellular virions are observed, displaying a cytopathic effect; b) They were identified Physio pathological processes using Masson's Trichrome Staining, detecting a predominance of the most advanced stages of Diffuse Alveolar Damage. Using Electron Microscopy, we found a profuse loss of alveolar epithelial cells in the lung. Confocal Microscopy was able to detect the presence of the SARS-CoV-2 nucleocapsid co-localized with key molecules such as Fibronectin, Vimentin, and PPARγ, together with NLRP3 Inflammasome in the brain cortical tissues. Conclusions. The isolation of the SARS-CoV-2 virus was performed for the first time in Cuba in nasopharyngeal exudates of patients with COVID-19 in the Vero E6 cell line. We identified for the first time novel cellular and pathophysiological processes involved in the infection of SARS-CoV-2, as well as potential therapeutic targets that reveal promising drugs to combat COVID-19 and its sequelae.

RESUMEN Introducción: La identificación del amplio tropismo del SARS-CoV-2 puede ser crítico para la patogénesis viral, por lo que es relevante conocer los mecanismos de infección e identificar nuevas dianas terapéuticas. Esta investigación consta de 2 bloques experimentales: aislamiento y caracterización del virus SARS-CoV-2 y patogénesis, tropismo, dianas moleculares y terapéuticas. Métodos: Se realizaron los siguientes procederes a) de la cavidad nasofaríngea de pacientes positivos al COVID-19, se obtuvo muestras clínicas que se inocularon en las células Vero E6 y se analizaron mediante microscopía electrónica de barrido a las 72 h postinfección; b) se obtuvo muestras de órganos viscerales de fallecidos por COVID-19: pulmón, hígado, riñón y cerebro. Los criocortes o cortes ultra finos se evaluaron mediante la tinción de tricrómica de masson, microscopía electrónica y la microscopia confocal. Resultados: Se obtuvo los siguientes resultados principales: a) se identificó un extenso daño en las células Vero E6 infectadas en las cuales se pudieron observar partículas de virus y viriones extracelulares, evidenciando un efecto citopático; b) se identificaron procesos fisiopatológicos: por tinción de tricrómica de Masson, se identificó un predominio de las etapas más avanzadas del daño alveolar difuso, al microscopio electrónico en pulmón, se evidenció la pérdida abundante de las células epiteliales alveolares. A través de microscopia confocal, se detectó la presencia de la nucleocápside del SARS-CoV-2 colocalizada con las moléculas claves tales como: fibronectina, vimentina, PPARγ conjuntamente con el inflamasoma NLRP3 en tejidos corticales cerebrales. Como conclusiones, en esta investigación: a) se realizó por primera vez el aislamiento del virus SARS-CoV-2 en Cuba en exudados nasofaríngeos de pacientes con COVID-19 en la línea celular Vero E6. b) Se identificó por primera vez; novedosos procesos celulares y fisiopatológicos involucrados en la infección del virus SARS-CoV-2 y potenciales dianas terapéuticas que revelan fármacos prometedores para combatir la COVID-19 y sus secuelas.

Caracterización palinológica de mieles del Valle de Mexicali, Baja California, México
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Alaniz-Gutiérrez, Luis

; Ail-Catzim, Carlos Enrique

; Villanueva-Gutiérrez, Rogel

; Delgadillo-Rodríguez, José

; Ortiz-Acosta, Martín Eduardo

; García-Moya, Edmundo

; Medina Cervantes, Tomás Salvador

Polibotánica

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2017, N. 43 Pages 255 - 283

Abstract: En Es | Text: En Es | PDF: En | PDF: Es

Abstract: The profitability of apiculture and the health of bee populations depend on landscapes rich in nectariferous and polleniferous flowers. Therefore it is clear that knowledge on local bee flora constitutes the basic information necessary to determine the apiculture potential of a region. Nevertheless, there is no research to date showing the floral sources of honey in the State of Baja California. Thus a melissopalynological study of the honey produced in the Valley of Mexicali, Baja California was undertaken to identify the main sources of pollen and nectar utilized by Apis mellifera and determine the botanical origin during the two principal harvest periods around year. The pollen of 52 honey samples was analyzed, these were collected from 13 apiaries on June and August of 2010 and of 2011. To identify the types of pollen, a reference collection was made containing 150 species pollen prepared via the acetolysis method. 78 types of pollen were identified, corresponding to 33 families. 65% of the honeys were unifloral, mainly from Tamarix spp., and Prosopis spp., and to a lesser extent of Pluchea sericea. Sysimbrium irio, Myrtaceae, Phoenixdactylifera, Coriandrum sativum, and Washingtonia filifera were found a single time. Prosopis spp. honeys were most abundant in June harvests, while Tamarix spp. honeys were the most common august harvests. Quantitative analysis allowed to allocate 52% of the samples on class III, 34% on class II, 8% on class V, 4% on class IV and 2% on class I. According to the analysis carried out, it is concluded that the main nectar sources used by A. mellifera in the Valley of Mexicali are Tamarix spp. Propsopis spp. and P. sericea, which are also the most representative elements of honeys.

Resumen: La rentabilidad de la producción apícola y las poblaciones saludables de abejas dependen de paisajes ricos en flores productoras de abundante néctar y polen. Por lo tanto, es claro que el conocimiento de la flora apícola constituye la información básica necesaria para determinar el potencial productivo de la apicultura en una región. En el estado de Baja California, se carece de este tipo de información. Por ello se realizó un estudio palinológico de las mieles producidas en el Valle de Mexicali, Baja California, para identificar los principales recursos nectaríferos y determinar el origen botánico durante dos periodos de cosecha anuales. Se analizaron 52 muestras de miel, recolectadas en 13 apiarios en junio y agosto de 2010 y 2011. Para identificar los tipos polínicos se conformó una colección de referencia que contiene polen acetolizado de 150 especies. Se identificaron 78 tipos polínicos, correspondientes a 33 familias. El 65% de las mieles fueron monoflorales, principalmente de Tamarix spp. (pino salado), Prosopis spp. (mezquite y tornillo) y en menor medida de Pluchea sericea (cachanilla), encontrándose una muestra de Sysimbrium irio (Mostacilla), Myrtaceae (eucaliptos y/o cepillo rojo), Phoenix dactylifera (palma datilera), Coriandrum sativum (cilantro), y Washingtonia filifera (palma de abanico) respectivamente. Las mieles de Prosopis spp. fueron abundantes en las cosechas de junio, mientras que las de Tamarix spp. predominaron en las cosechas de agosto. Los análisis cuantitativos, permitieron ubicar el 52% de las muestras en la clase III, el 34% enlaclaseII, el 8% en la clase V, el 4% en la clase IV y el 2% en la I. Deacuerdo a los estudios realizados se concluye que los principales recursos nectaríferos usados por Apis mellifera en el Valle de Mexicali son Tamarix spp. Propsopis spp. y P. sericea, los cuales a su vez constituyen los elementos más representativos de las mieles.

https://doi.org/10.18387/polibotanica.43.12 1997 downloads

Growth inhibition of Staphylococcus aureus and escherichia coli strains by neutralizing IgY antibodies from ostrich egg yolk
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Tobias, Fernando Luiz

; Garcia, Luize Néli Nunes

; Kanashiro, Milton Masahiko

; Medina-Acosta, Enrique

; Brom-de-Luna, João Gato

; Almeida, Claudia Maria Costa de

; Azevedo Junior, Romildo Rocha

; Lemos, Môsar

; Vieira-da-Motta, Olney

Brazilian Journal of Microbiology

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jun 2012, Volume 43 N. 2 Pages 544 - 551

Abstract: En | Text: En | PDF: En

Ostrich raising around the world have some key factors and farming profit depend largely on information and ability of farmers to rear these animals. Non fertilized eggs from ostriches are discharged in the reproduction season. Staphylococcus aureus and Escherichia coli are microorganisms involved in animal and human diseases. In order to optimize the use of sub products of ostrich raising, non fertilized eggs of four selected birds were utilized for development of polyclonal IgY antibodies. The birds were immunized (200ug/animal) with purified recombinant staphylococcal enterotoxin C (recSEC) and synthetic recRAP, both derived from S. aureus, and recBFPA and recEspB involved in E. coli pathogenicity, diluted in FCA injected in the braquial muscle. Two subsequent immunization steps with 21 days intervals were repeated in 0,85% saline in FIA. Blood and eggs samples were collected before and after immunization steps. Egg yolk immunoglobulins were purified by precipitation with 19% sodium sulfate and 20% ammonium sulphate methodologies. Purified IgY 50µL aliquots were incubated in 850µL BHI broth containing 50µL inoculums of five strains of S. aureus and five strains of E.coli during four hours at 37ºC. Growth inhibition was evaluated followed by photometry reading (DO550nm). Egg yolk IgY preparation from hiperimmunized birds contained antibodies that inhibited significantly (p<0,05) growth of strains tested. Potential use of ostrich IgY polyclonal antibodies as a diagnostic and therapeutic tool is proposed for diseased animals.

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Cáncer de cuello uterino: Consenso nacional para el diagnóstico y tratamiento 2010
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Medina E, Francisco

; Sánchez-Lander, Jorge

; Calderaro, Franco

; Borges, Alfredo

; Rennola, Antonieta

; Bermúdez, Cecilia

; García de Barriola, Victoria

; Ruiz, Ángela

; Naranjo de Gómez, Miriam

; Reigosa, Aldo

; López Loyo, Enrique

; Guillén, Nelson

; Vásquez, Jesús

; Di Giampietro, Loretta

; López, Nora

; Bolívar, Luisa

; Briceño, Wilmar

; Acosta Garnier, Humberto

; Blanch, Ricardo

; González, Claudia

; Ávila, Maira

; González Blanco, Mireya

; Oberto, José

; Díaz Bruzual, Alfredo

; Vici, Gastón

; Manzo, Aisa

; Fernández, Luis

; Peña, Orlando

; Moreira, Erick

; Rezic, Marko

; Castillo Romero, Jorge A

; Lion, Lorena

; Barbosa, Doris

; Aguilera, María

; Capote Negrín, Luis

; Pérez, Ángel

; Lugo, José Ángel

; Rodríguez, Ivo

; Anzola, Félix

; Vento, Giovanni

; Fuentes, María Belén

; Nass, Ingrid

; Benítez, María Mercedes

; Agüero, Belkys

; Medina, Ivonne

; Palacios, Priscila

; Umbria, Carmen

; Rebolledo, Morella

; Chacón, Marco

; Bonilla, Patricia

; Montañez, Ángela

; García, María Belén

; Vázquez, Belkis

; Márquez, Ralyma

; Hernández, Yoly

; Pérez Rivas, Janett

; Lozada, Marnie

; Melet, Argelia

Revista Venezolana de Oncología

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jun 2011, Volume 23 N. 2 Pages 102 - 129

Abstract: Es | Text: Es | PDF: Es

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Desafios operacionais persistentes determinam a não redução da transmissão materno-infantil do HIV
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Fernandes, Regina Célia de Souza Campos

; Ribas, Gustavo Fernandes

; Silva, Danielli Pires e

; Gomes, Alexandre Machado

; Medina-Acosta, Enrique

Jornal de Pediatria

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dec 2010, Volume 86 N. 6 Pages 503 - 508

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO: Determinar os obstáculos à efetiva redução da transmissão materno-infantil do HIV em Campos dos Goytacazes (RJ). MÉTODOS: Estudo de coorte, com acompanhamento médico de gestantes com diagnóstico confirmado de infecção pelo HIV e de seus bebês, atendidos no Serviço Municipal de Atendimento Especializado de Doenças Sexualmente Transmissíveis e AIDS, no período de janeiro de 2004 a abril de 2007; coleta de informações relativas às variáveis de exposição e desfecho a partir dos prontuários; determinação de frequências das variáveis e análise bivariada entre os fatores de exposição e o desfecho; cálculo de riscos relativos de transmissão materno-infantil do HIV associados às variáveis de exposição, com intervalo de confiança de 95%; e avaliação de significância estatística de associações de risco. RESULTADOS: Foram acompanhados 78 binômios, sendo de 7,7% a taxa de transmissão materno-infantil do HIV. As variáveis que mostraram associações estatisticamente significantes com a transmissão materno-infantil do HIV foram: a não utilização de antirretrovirais para profilaxia ou tratamento na gestação (RR = 21,00; IC95% 2,64-166,74, p = 0,001); e o diagnóstico materno após a gestação (RR = 6,80; IC95% 1,59-29,17, p = 0,025). Novas gestações em mulheres com outros filhos já expostos ao HIV ocorreram em 19,12% dos casos. CONCLUSÕES: Não houve redução da taxa de transmissão materno-infantil do HIV no período de 2004-2007 em relação ao triênio anterior. Foram caracterizados como obstáculos determinantes desta não redução: a baixa cobertura pela testagem anti-HIV no pré-natal, impossibilitando tratamento ou profilaxia materna eficiente; e o uso incorreto do teste rápido na admissão para o parto.

OBJECTIVE: To determine impediments to the effective reduction of maternal-infant transmission of HIV in the municipality of Campos dos Goytacazes, RJ, Brazil. METHODS: This is a cohort study, with medical follow-up, of pregnant women with confirmed diagnosis of HIV infection, and their infant children, assisted at the Municipal Specialized Service of Sexually Transmitted Diseases/AIDS from January 2004 to April 2007. Information regarding exposure and outcome variables was collected from their medical records. Frequencies of variables were determined and bivariate analysis performed for exposure factors and transmission of HIV. Relative risks of HIV transmission associated with exposure variables were calculated using 95% confidence intervals. Statistical significance of risk associations was evaluated. RESULTS: Seventy-eight mother-child pairs were studied; the rate of maternal-infant transmission of HIV was 7.7%. Variables showing significant association with maternal-infant transmission of HIV were the non-utilization of antiretrovirals for prophylaxis or treatment during pregnancy (RR = 21.00; 95%CI 2.64-166.74, p = 0.001) and diagnosis of maternal disease after pregnancy (RR = 6.80; 95%CI 1.59-29.17, p = 0.025). New pregnancies in women with other children also exposed to HIV occurred in 19.12% (15/78) of cases. CONCLUSIONS: There was no reduction in the rate of maternal-infant transmission of HIV in the period 2004-2007 in relation to the preceding triennium. The following were recognized as impediments to the effective reduction of maternal-infant transmission of HIV: low prenatal screening coverage of maternal HIV infection, impairing maternal treatment or prophylaxis; and the incorrect use of the rapid screening test at admission for delivery.

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Vertical transmission of HIV and toxoplasma by reactivation in a chronically infected woman
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Fernandes, Regina Célia de Souza Campos

; Vasconcellos, Vanessa Perez

; Araújo, Luciana Cordeiro de

; Medina-Acosta, Enrique

Brazilian Journal of Infectious Diseases

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feb 2009, Volume 13 N. 1 Pages 70 - 71

Abstract: En | Text: En | PDF: En

3157 downloads

Propiedades termo-ópticas y procesos de transferencia de energía en vidrios con Erbio preparados vía sol-gel
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Medina Monares, Joel

; Gutiérrez Acosta, Keren-Hapuc

; Duarte Moller, Alberto

; Apolinar Iribe, Alejandro

; Álvarez R., Mario Enrique

; Castillo, Santos Jesús

Superficies y vacío

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2007, Volume 20 N. 4 Pages 10 - 13

Abstract: En Es | Text: En Es | PDF: En | PDF: Es

We present a study of the optical properties of glass matrices with Erbium trivalent impurities, which were synthesized by Sol-Gel technology. The experiments of optical absorption, emission and excitation were carried out at room temperature between range 300 to 1000 nm. It identifies and discusses the existence of a of radiative energy transfer process between the network and the Er3+ ions. It also presents the characteristics of thermo-optic on condition of resonant absorption of the ion Er3+, for different concentrations of Erbium. The effect of Thermal Lens noticed is discussed in terms of theoretical model.

Se presenta un estudio de las propiedades ópticas de matrices vítreas con impurezas de erbio trivalente, que fueron sintetizadas mediante la técnica sol-gel. Los experimentos de absorción óptica, de emisión y de excitación se realizaron a temperatura ambiente en el intervalo de 300 a 1000 nm. Se identifica y discute la existencia de un proceso de transferencia de energía radiativa entre la red → Er3+ y las características de termo-ópticas bajo la condición resonante a la absorción del ion Er3+, para diferentes concentraciones de erbio. Se observo el efecto de Lente Térmica, misma que fue analizada en términos de un modelo teórico sencillo que permite explicar los resultados experimentales.

1055 downloads

O desafio da prevenção da transmissão vertical do HIV no Município de Campos dos Goytacazes, Rio de Janeiro, Brasil
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Fernandes, Regina Célia de Souza Campos

; Araújo, Luciana Cordeiro de

; Medina-Acosta, Enrique

Cadernos de Saúde Pública

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aug 2005, Volume 21 N. 4 Pages 1153 - 1159

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

A transmissão vertical do HIV continua sendo um importante desafio para a Saúde Pública. A partir de 1996 se iniciou no Brasil o uso da profilaxia com Zidovudina durante a gestação, no momento do parto e para o recém-nascido, com redução da transmissão para menos de 5,0% em muitos serviços. Desde 1999, o Programa Municipal de DST/AIDS de Campos dos Goytacazes (população: 416.441), Rio de Janeiro, atua na prevenção da transmissão perinatal do HIV. Merecem destaque o moderado aumento da captação de gestantes HIV positivas do município (de 28,0% para 36,0% no período 2001-2003), por meio da maior oferta do teste de HIV no pré-natal e o maior conhecimento que as gestantes demonstram sobre a transmissão materno-infantil do HIV. Persistem como desafios a extensão da testagem de HIV a toda a população de gestantes e, no caso das não testadas, o uso do teste rápido para diagnóstico no momento do parto. Com isso, se conseguiria modificar o panorama atual em que apenas 6,8% das crianças infectadas e em acompanhamento tiveram acesso à profilaxia da transmissão materno-infantil do HIV.

Prevention of mother-to-child HIV transmission is an emerging public health challenge. In Brazil, preventive prophylaxis with AZT during pregnancy, at delivery, and for newborns was introduced in 1996, with a marked reduction in transmission (less than 5.0% nationwide). Since 1999 the Municipal STD/AIDS Program in Campos dos Goytacazes, Rio de Janeiro State (population = 416,441) has acted in the prevention of vertical transmission. In 2001/2003, there was a moderately increased implementation (from 28.0% to 36.0%) of HIV screening services during pregnancy, and more pregnant women were knowledgeable about vertical HIV transmission. Remaining challenges are to expand HIV screening to pregnant women at large and to effectively introduce rapid diagnostic tests for HIV in the delivery room. Such actions are expected to change the current situation, in which only 6.8% of infected children under observation had access to prophylaxis for vertical HIV transmission.

5772 downloads

O desafio da prevenção da transmissão vertical do HIV no Município de Campos dos Goytacazes, Rio de Janeiro, Brasil
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Fernandes, Regina Célia de Souza Campos

; Araújo, Luciana Cordeiro de

; Medina-Acosta, Enrique

Cadernos de Saúde Pública

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aug 2005, Volume 21 N. 4 Pages 1153 - 1159

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

6121 downloads

10.

Genetic Data Showing Evolutionary Links between Leishmania and Endotrypanum
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Cupolillo, Elisa

; Pereira, Luiza OR

; Fernandes, Octávio

; Catanho, Marcos P

; Pereira, Júlio C

; Medina-Acosta, Enrique

; Grimaldi Jr, Gabriel

Memórias do Instituto Oswaldo Cruz

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sep 1998, Volume 93 N. 5 Pages 677 - 683

Abstract: En | Text: En | PDF: En

Striking similarities at the morphological, molecular and biological levels exist between many trypanosomatids isolated from sylvatic insects and/or vertebrate reservoir hosts that make the identification of medically important parasites demanding. Some molecular data have pointed to the relationship between some Leishmania species and Endotrypanum, which has an important epidemiological significance and can be helpful to understand the evolution of those parasites. In this study, we have demonstrated a close genetic relationship between Endotrypanum and two new leishmanial species, L. (V.) colombiensis and L. (V.) equatorensis. We have used (a) numerical zymotaxonomy and (b) the variability of the internal transcribed spacers of the rRNA genes to examine relationships in this group. The evolutionary trees obtained revealed high genetic similarity between L. (V.) colombiensis, L. (V.) equatorensis and Endotrypanum, forming a tight cluster of parasites. Based on further results of (c) minicircle kDNA heterogeneity analysis and (d) measurement of the sialidase activity these parasites were also grouped together.

1893 downloads Cited 1 time in SciELO

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