Raquel E. Ocazionez

Industrial University of Santander

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Elena E. Stashenko

Industrial University of Santander

Sergio Yebrail Gómez

University of Santander

Fabián Cortés-Mancera

Instituto Tecnológico Metropolitano

Luis Villar

Industrial University of Santander

Paola Rondón-Villarreal

University of Santander

Elizabeth Quintero-Rueda

Industrial University of Santander

Jorge Luı́s Fuentes

Industrial University of Santander

M. Alarcon

Hospital de Clínicas

Lina Silva-Trujillo

Industrial University of Santander

María C. Flechas

Industrial University of Santander

Cooperative Institutions

Industrial University of Santander

University of Cartagena

Harvard University

Universidad Nacional Autónoma de México

University of Santander

Universidad Autónoma de Bucaramanga

Massachusetts Institute of Technology

Autonomous University of Tlaxcala

Pontificia Universidad Católica de Chile

Universidad de Antioquia

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In vitro Safety Assessment of Extracts and Compounds From Plants as Sunscreen Ingredients

International Journal of Toxicology (2024)

Silvia Juliana Flórez González Elena E. Stashenko Raquel E. Ocazionez M. Pilar Vinardell Jorge Luı́s Fuentes

This work investigated the safety of extracts obtained from plants growing in Colombia, which have previously shown UV-filter/antigenotoxic properties. The compounds in plant extracts obtained by the supercritical fluid (CO 2 ) extraction method were identified using gas chromatography coupled to mass spectrometry (GC/MS) analysis. Cytotoxicity measured as cytotoxic concentration 50% (CC 50 ) and genotoxicity of the plant extracts and some compounds were studied in human fibroblasts using the trypan blue exclusion assay and the Comet assay, respectively. The extracts from Pipper eriopodon and Salvia aratocensis species and the compound trans-β-caryophyllene were clearly cytotoxic to human fibroblasts. Conversely, Achyrocline satureioides, Chromolaena pellia, and Lippia origanoides extracts were relatively less cytotoxic with CC 50 values of 173, 184, and 89 μg/mL, respectively. The C. pellia and L. origanoides extracts produced some degree of DNA breaks at cytotoxic concentrations. The cytotoxicity of the studied compounds was as follows, with lower CC 50 values representing the most cytotoxic compounds: resveratrol (91 μM) > pinocembrin (144 μM) > quercetin (222 μM) > titanium dioxide (704 μM). Quercetin was unique among the compounds assayed in being genotoxic to human fibroblasts. Our work indicates that phytochemicals can be cytotoxic and genotoxic, demonstrating the need to establish safe concentrations of these extracts for their potential use in cosmetics.

Comet Assay

Trypan blue

Isoorientin

10.1177/10915818231225661

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Evaluation of in vitro Antiviral Activity of Essential Oil Compounds Against Dengue Virus

Pharmacognosy Journal (2017)

María C. Flechas Raquel E. Ocazionez Elena E. Stashenko

Introduction: There are not effective drugs available for treatment of dengue fever despite intensive research on synthetic inhibitors. The search for active phytochemicals could serve for the discovery of new drugs. This study aims to evaluate the antiviral activity in vitro of compounds found in essential oils from medicinal plants. Materials and Methods: Nine synthetic-derived essential oil compounds were evaluated. Antiviral effect was screened measuring the reduction of viral NS1 and E proteins in HepG-2 and Vero cells. Results: β-Caryophyllene was identified as the most active compound, it reduced the virus serotype-2 replication in HepG-2 cells at IC50 of 22 ± 5.6 μM, and blocked replication of all four serotypes in Vero cells at IC50 between 8.0 μM and 15.0 μM. The selectivity indexes were between 5.3 and 10. According to results from time-in-addition assays, the antiviral effect of β-caryophyllene appears to be associated with interruption of early steps of the virus life cycle. Citral revealed modest antiviral effect, it reduced the virus serotype-2 (IC50 of 31 ± 4.5 μM) replication but not the other three serotypes. Seven terpenes did not reveal antiviral activity at maximum concentration of 30 μM. Conclusion: Research on compounds found in essential oils can contribute to the drug discovery effort for dengue. β-Caryophyllene could serve as a starting point.

Vero cell

IC50

Antiviral drug

10.5530/pj.2018.1.11

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Comparative Study on In Vitro Activities of Citral, Limonene and Essential Oils from Lippia citriodora and L. alba on Yellow Fever Virus

Natural Product Communications (2013)

Luz Ángela Gómez Elena E. Stashenko Raquel E. Ocazionez

The aim of this study was to compare the antiviral activities in vitro of citral, limonene and essential oils (EOs) from Lippia citriodora and L. alba on the replication of yellow fever virus (YFV). Citral and EOs were active before and after virus adsorption on cells; IC50 values were between 4.3 and 25 microg/mL and SI ranged from 1.1 to 10.8. Results indicate that citral could contribute to the antiviral activity of the L. citriodora EO. Limonene was not active and seemed to play an insignificant role in the antiviral activity of the examined EOs.

Citral

Lippia

Citronellal

10.1177/1934578x1300800230

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Anticuerpos Neutralizantes contra el Virus de la Fiebre Amarilla 17 D en Colombianos Vacunados y no Vacunados con Inmunidad a Dengue Yellow fever virus 17D neutralising antibodies in vaccinated Colombian people and unvaccinated ones having immunity against dengue

Sergio Yebrail Gómez Raquel E. Ocazionez

RESUMENObjetivo Determinar la frecuencia de titulo protector de anticuerpos neutralizantescontra el virus de la fiebre amarilla (AN-VFA a titulo >1:10) en colombianos vacuna-dos con la cepa 17 D y conocer la magnitud de neutralizacion del VFA por anticuerposcontra dengue. Metodologia Se colecto suero de 100 individuos con vacuna documentada porcarne y de 116 residentes en municipios de Norte de Santander afectados por elbrote en 2002-2003, quienes informaron haber sido vacunados. Se incluyeron sue-ros de individuos no vacunados con (n=61) y sin (n=16) anticuerpos contra dengue.Todos los sueros se analizaron por la prueba de neutralizacion para VFA por 75 %de reduccion de placa. Resultados AN-VFA a titulo >1:10 se encontraron en 90 % de vacunados con carney sin variacion aparente en relacion con edad. Al contrario, hubo correlacion entredisminucion de la frecuencia de titulo protector de anticuerpos e incremento deltiempo de inmunizacion (r=0,95; p=0,04). En residentes de Norte de Santander, AN-VFA a titulo >1:10 se encontraron en 92,6 % adultos y 69 % ninos. El VFA fueneutralizado (52 -100 %) por sueros de inmunes a dengue mas eficientemente quepor sueros de no inmunes (p<0.001).

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Surveillance of the Dengue based on the laboratory: differences in the number of cases and virus isolated according to the gathering of the serum and serologic tests

Colombia medica (2005)

Raquel E. Ocazionez Fabián Cortés-Mancera Luis Villar

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Virucidal activity of Colombian Lippia essential oils on dengue virus replication in vitro

Memórias do Instituto Oswaldo Cruz (2010)

Raquel E. Ocazionez Rocío Meneses Flor Ángela Torres Elena E. Stashenko

The inhibitory effect of Lippia alba and Lippia citriodora essential oils on dengue virus serotypes replication in vitro was investigated. The cytotoxicity (CC50) was evaluated by the MTT assay and the mode of viral inhibitory effect was investigated with a plaque reduction assay. The virus was treated with the essential oil for 2 h at 37ºC before cell adsorption and experiments were conducted to evaluate inhibition of untreated-virus replication in the presence of oil. Antiviral activity was defined as the concentration of essential oil that caused 50% reduction of the virus plaque number (IC50). L. alba oil resulted in less cytotoxicity than L. citriodora oil (CC50: 139.5 vs. 57.6 μg/mL). Virus plaque reduction for all four dengue serotypes was observed by treatment of the virus before adsorption on cell. The IC50 values for L. alba oil were between 0.4-32.6 μg/mL and between 1.9-33.7 μg/mL for L. citriodora oil. No viral inhibitory effect was observed by addition of the essential oil after virus adsorption. The inhibitory effect of the essential oil seems to cause direct virus inactivation before adsorption on host cell.

Virus quantification

Lippia

MTT assay

IC50

10.1590/s0074-02762010000300010

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Comparative Anti-dengue Activities of Ethanolic and Supercritical Extracts of Lippia origanoides Kunth: in-vitro and in-silico Analyses

Records of Natural Products (2023)

Elizabeth Quintero-Rueda Sindi Alejandra Velandia Raquel E. Ocazionez Elena E. Stashenko Paola Rondón-Villarreal

Despite intense research efforts, no approved effective therapies are available for dengue treatment.Standardized plant extracts could serve as potential candidates for developing plant-based treatments for dengue.We have compared the anti-dengue potential of ethanolic (EtOH) and supercritical (SCO2) extracts of Lippia origanoides Kunth (Verbenacea).The mode of antiviral action and the effect on the release of IL-β from DENV-1-stimulated macrophages (PMA/THP-1) were evaluated.In silico analyses were performed to predict molecular interactions between flavonoids identified in the UHPLC-ESI+-Orbitrap-MS analysis and target proteins.The EtOH extract showed a strong antiviral effect (IC50, 22 and 17 µg/mL; SI, 20 and 23) before and during DENV-1 adsorption to liver (HepG-2) cells.SCO2 showed a weak antiviral effect.EtOH, but not SCO2, reduced the level of IL-1β released from PMA/THP-1 cells by 45-55%.Flavonoid glycosides were identified in EtOH but not in SCO2.Seventeen flavonoids were predicted to bind to DENV E, GRP78, CLEC5A, and NLRP3 proteins involved in DENV replication and IL-1β production.The data provide a first step towards defining the potential of L. origanoides extract as a candidate for developing phytotherapeutics for dengue.

Lippia

10.25135/rnp.401.2303.2720

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Dengue in Santander State, Colombia: fluctuations in the prevalence of virus serotypes are linked to dengue incidence and genetic diversity of the circulating viruses

Tropical Medicine & International Health (2019)

María Fernanda Carreño Cinthy L. Jiménez-Silva Luz Aída Rey-Caro Sergio Conde‐Ocazionez M. Alarcon

To investigate the link between fluctuations in the prevalence of dengue virus (DENV) serotypes and the number of dengue cases in the metropolitan area of Bucaramanga, Santander State, Colombia, in the 2007-2010 and 2014-2017 periods.Viruses were isolated from febrile patient samples by direct application to C6/36-HT cells and typed using monoclonal antibodies. We performed autocorrelation and cross-correlation analyses to determine whether fluctuations in the prevalence of DENV serotypes and dengue cases were correlated. Full envelope (E) gene sequences were employed to examine the genetic diversity of serotypes circulating by using a phylogenetic approach.All four dengue virus serotypes were detected. DENV-1 was the dominant serotype in both periods followed by DENV-3 or DENV-2 depending on the period; DENV-4 was the least prevalent virus in both periods. Cross-correlation analyses suggest a temporal relation between the fluctuations in the prevalence of DENV serotypes, which were almost simultaneous (lag = 0) or related to recent past fluctuations (lag > 1.0) in the number of dengue cases. Data suggest that a sustained predominance of DENV-1, an increase of the DENV-4 prevalence, and a switch from DENV-3 to DENV-2 could be linked to an outbreak. Circulating viruses were grouped into Genotype V, Asia/American III and II for DENV-1, -2, -3 and -4, respectively; intragenotypic diversity was detected.The present work highlights the need of comprehensive studies on dynamics of DENV in Colombia to understand transmission of dengue and evaluate the effectiveness of a vaccination programme.Etudier le lien entre les fluctuations de la prévalence des sérotypes du virus de la dengue (DENV) et le nombre de cas de dengue dans la région métropolitaine de Bucaramanga, dans l’Etat de Santander, en Colombie, au cours des périodes 2007-2010 et 2014-2017. MÉTHODE: Les virus ont été isolés à partir d'échantillons de patients fébriles par application directe sur des cellules C6/36-HT et typés à l'aide d'anticorps monoclonaux. Nous avons effectué des analyses d'autocorrélation et de corrélation croisée afin de déterminer si les fluctuations de la prévalence des sérotypes du DENV et des cas de dengue étaient corrélées. Des séquences de gènes d’enveloppe complète (E) ont été utilisées pour examiner la diversité génétique des sérotypes en circulation en utilisant une approche phylogénétique. RÉSULTATS: Tous les quatre sérotypes du virus de la dengue ont été détectés. DENV-1 était le sérotype dominant dans les deux périodes, suivi de DENV-3 ou DENV-2, selon la période; le virus DENV-4 était le moins prévalent au cours des deux périodes. Les analyses de corrélation croisée suggèrent une relation temporelle entre les fluctuations de la prévalence des sérotypes de DENV, qui étaient presque simultanées (lag = 0) ou liées aux fluctuations passées récentes (lag > 1,0) du nombre de cas de dengue. Les données suggèrent qu'une prédominance durable de DENV-1, qu’une augmentation de la prévalence de DENV-4 et qu’un passage de DENV-3 à DENV-2 pourraient être liés à une éclosion. Les virus en circulation ont été regroupés dans les génotypes V, Asie/Amérique III et II pour DENV-1, -2, -3 et -4, respectivement; une diversité intra-génotypique a été détectée.Le présent travail souligne la nécessité d’études approfondies sur la dynamique du DENV en Colombie afin de comprendre la transmission de la dengue et évaluer l’efficacité d’un programme de vaccination.

10.1111/tmi.13311

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Actividad antiproliferativa de aceites esenciales de plantas cultivadas en Colombia

Acta Biológica Colombiana (2018)

Sindi Alejandra Velandia Elizabeth Quintero Elena E. Stashenko Raquel E. Ocazionez

Colombia posee gran diversidad de plantas medicinales, pero pocas han sido objeto de investigación. En este trabajo se evaluó la actividad antiproliferativa de aceites esenciales obtenidos por hidrodestilación asistida por microondas. Se analizaron 15 muestras de 11 especies en ensayos del MTT en células cancerosas MCF-7, HeLa y HepG-2 y se incluyeron células normales humanas (HEK293) y de animales (Vero y BF16F10) para evaluar selectividad. La composición química de muestras activas se determinó por cromatografía de gases acoplada a espectrometría de masas (GC-MS). Aceites esenciales de cuatro especies mostraron actividad antiproliferativa (CI50: 50 μg/mL) en células HeLa y/o MCF-7, en el siguiente rango (índice de selectividad en paréntesis): Piper cumanense H.B.K. (4,7) > Piper subflavum var. espejuelanum C.DC (3,9) > Salvia officinalis L. (3,6) > Piper eriopodom (Miq.) C. DC. (3,1). Ninguna muestra fue activa en células HepG-2. El análisis por CG-MS identificó β-cariofileno, α-copaeno, β-pineno, α-pineno y linalol como componentes mayoritarios. Los aceites esenciales activos pueden ser punto de partida para desarrollo de medicamentos herbales para cuidado paliativo del cáncer.

10.15446/abc.v23n2.67394

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Citations (8)

Comparative Virucidal Activities of Essential Oils and Alcohol-Based Solutions against Enveloped Virus Surrogates: In Vitro and In Silico Analyses

Molecules (2023)

Valentina Parra-Acevedo Raquel E. Ocazionez Elena E. Stashenko Lina Silva-Trujillo Paola Rondón-Villarreal

The large-scale use of alcohol (OH)-based disinfectants to control pathogenic viruses is of great concern because of their side effects on humans and harmful impact on the environment. There is an urgent need to develop safe and environmentally friendly disinfectants. Essential oils (EOs) are generally recognized as safe (GRAS) by the FDA, and many exhibit strong antiviral efficacy against pathogenic human enveloped viruses. The present study investigated the virucidal disinfectant activity of solutions containing EO and OH against DENV-2 and CHIKV, which were used as surrogate viruses for human pathogenic enveloped viruses. The quantitative suspension test was used. A solution containing 12% EO + 10% OH reduced > 4.0 log10 TCID50 (100% reduction) of both viruses within 1 min of exposure. In addition, solutions containing 12% EO and 3% EO without OH reduced > 4.0 log10 TCID50 of both viruses after 10 min and 30 min of exposure, respectively. The binding affinities of 42 EO compounds and viral envelope proteins were investigated through docking analyses. Sesquiterpene showed the highest binding affinities (from −6.7 to −8.0 kcal/mol) with DENV-2 E and CHIKV E1-E2-E3 proteins. The data provide a first step toward defining the potential of EOs as disinfectants.

Disinfectant

Binding affinities

10.3390/molecules28104156

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Citations (4)