Cervicitis necrótica por co-infección de virus herpes simplex 2 y Mycoplasma genitalium / Necrotic cervicitis for co-infection of herpes simplex virus 2 and Mycoplasma genitalium

Resumen Las cervicitis es una condición frecuente causada principalmente por agentes de transmisión sexual. Su presentación clínica varía desde cuadros asintomáticos hasta procesos inflamatorios extensos, que incluso asemejan un tumor maligno. Presentamos el caso de una adolescente que presentó úlceras genitales, síntomas generales y cérvix necrótico con aspecto tumoral. Los estudios de laboratorio confirmaron una co-infección por virus herpes simplex 2 (HSV-2) y Mycoplasma genitalium. El estudio histológico descartó una neo- plasia. Evolucionó favorablemente al tratamiento antimicrobiano, con recuperación progresiva del aspecto del cérvix. La cervicitis en raras ocasiones se presenta con compromiso necrótico. La co-infección por HSV-2 y M. genitalium, en este caso, pudo ser el determinante del daño cervical y la necrosis. Una evaluación acuciosa y estudio con exámenes diagnósticos de alta sensibilidad y especificidad permitieron hacer un diagnóstico y tratamiento adecuado.

Abstract Cervicitis is a frequent condition caused mainly by sexually trans- mitted agents. The clinical spectrum varies from absence of symptoms to extensive inflammatory processes that may simulate a malignant neoplasm. We present a clinical case of an adolescent with genital ulcers and systemic disease. Speculoscopy revealed a tumoral-looking cervix. Laboratory studies confirm infection with herpes simplex virus 2 (HSV-2) and Mycoplasma genitalium, together with a histological study that ruled out neoplasia. It progresses favorably to antimicrobial treatment, with recovery of the appearance of the cervix. Cervicitis rarely presents with necrotic involvement. Co-infection with HSV-2 and M. genitalium infection may have been the determinant of cervical damage and the necrotic appearance. A thorough evaluation and study with highly sensitive and specific diagnostic tests allowed an adequate diagnosis and treatment.

Diagnóstico molecular de infección por virus herpes simplex tipo 1 en tejido aterosclerótico humano / Molecular diagnosis of infection by herpes simplex type 1 virus in human aterosclerótico tissue

Introducción:Las enfermedades del sistema circulatorio representan uno de los mayores problemas de salud pública a nivel mundial, nacional y regional. Elmecanismo patogénico que subyace esta patología es la aterosclerosis. Existenvarios factores que favorecen la etiopatogeniade la lesión aterosclerótica.Las infecciones, juegan un papel importante.La infección por el Virus del Herpes Simplexse ha considerado como un factor de riesgo emergente. Objetivo:Realizar diagnósticomolecular de infección porVirus Herpes Simplex tipo 1 y tipo 2en tejido aterosclerótico humano.Método:Se realizó extracción de ADN viral a partir de ateromas usando el kit comercial PureGenomeTM Tissue DNA Extraction.La amplificación delmaterial genético viralse realizó porPCR en tiempo real (qPCR) con el kit comercial "Human Herpes Virus 2 (Herpes simplex type 2)UL36 region genesig Standard Kit y Human Herpes Virus 1 (Herpes simplex type 1) Capsid assembly and DNA maturation gene. Genesig Standard Kit".Resultados:En total se obtuvieron 102 muestras de ateromas, extraídas de diferentes fuentes anatómicas. Tresmuestras resultaronpositivas para VHS tipo 1(3/102).Ninguna muestra evidenció material genético para VHS tipo 2 (0/102). Conclusión:La etiopatogenia de la aterosclerosis es un proceso altamente complejo.Los virus juegan un papel importante, en especial la infección por Virus del herpes simplex tipo 1. La infección por estevirus genera cambios a nivel de las células vasculares y no vasculares, favoreciendo el acumulo de lipoproteínas de baja densidad químicamente oxidadas, importantespara la aterogénesis

Introduction:Diseases of the circulatory system represent one of the greatest public health problems worldwide, nationally and regionally. The pathogenic mechanism that underlies this pathology is atherosclerosis. There are several factors that favor the etiopathogeny of the atherosclerotic lesion. Infections play an important role. Infection with Herpes Simplex Virus has been considered as an emerging risk factor. Objective: To perform molecular diagnosis of infection by Herpes Simplex virus type 1 and type 2 in human atherosclerotic tissue. Method:Viral DNA extraction was performed from atheromas using the commercial PureGenomeTM Tissue DNA Extraction kit. The amplification of the viral genetic material was performed by real-time PCR (qPCR) with the commercial kit "Human Herpes Virus 2 (Herpes simplex type 2) UL36 region genesig Standard Kit and Human Herpes Virus 1 (Herpes simplex type 1) Capsid assembly and DNA maturation gene Genesig Standard Kit ". Results:A total of 102 samples of atheromas were obtained, extracted from different anatomicalsources. Three samples were positive for HSV type 1 (3/102). No sample showed genetic material for HSV type 2 (0/102). Conclusion:The etiopathogenesis of atherosclerosis is a highly complex process. Viruses play an important role, especially the infection by Herpes simplex virus type 1. The infection by this virus generates changes at the level of vascular and non-vascular cells, favoring the accumulation of chemically oxidized low density lipoproteins, important for the atherogenesis

Eritema multiforme asociado a herpes virus: reporte de caso / Erythema multiform associated with herpes virus: case report

El eritema multiforme (EM) es una reacción de hipersensibilidad que afecta a la piel y / oa las membranas mucosas, en respuesta a infecciones, fármacos u otras comorbilidades. La presentación de EM puede ser Menor (EM leve, caracterizada por presentar lesiones diana) o Mayor (EM severa, que también ocurre con afectación de las membranas mucosas). Se presenta el caso de un paciente varón de 36 años de edad que presenta una emergencia de HNAL con fiebre, malestar, prurito y lesiones dolorosas en la cavidad oral, lesiones múltiples en miembros superiores e inferiores, pene y región perianal. El diagnóstico fue EM Major. Las lesiones ulcerosas en la cavidad oral y las lesiones diana en las manos fueron las más destacadas.

Erythema multiforme (EM) is a hypersensitivity reaction that affects the skin and/or mucous membranes, in response to infections, drugs, or other comorbidities. Presentation of EM can be Minor (Mild EM, characterized for presenting target lesions) or Major (Severe EM, that occurs also with mucous membranes involvement). The case of a 36 year old male patient presenting to emergency of HNAL with fever, malaise, itching and painful ulcer type lesions in oral cavity, multiple target lesions in upper and lower limbs, penis, and perianal region is reported. The diagnosis was EM Major. Ulcerous lesions in oral cavity and target lesions in hands were the most salient.

Infecciones virales en neonatos febriles con sepsis clínica. Hospital del Niño. Panamá Diciembre 2013-2014

Introducción: La fiebre en neonatos está comúnmente asociada a infecciones virales. La infección por Virus Herpes Simplex (VHS) en neonatos conlleva una alta morbi-mortalidad y secuelas importantes a largo plazo. En los últimos años esta infección ha presentado un aumento en su incidencia. El enterovirus, es una causa bien conocida de fiebre y meningitis aséptica en neonatos. La infección por citomegalovirus en neonatos es la infección viral congénita más común, afectando 0,2% a 2,5% de todos los nacidos vivos. La prueba de PCR ha sido utilizada con éxito para la identificación de ADN de CMV, virus Herpes y Enterovirus en recién nacidos. El objetivo de nuestro estudio fue determinar la frecuencia de infección virales en neonatos febriles, con sospecha de sepsis en el período de estudio en el Hospital del Niño. Materiales y métodos: Se evaluó todo neonato febril que ingreso a la sala de Neonatología u otra sala de hospitalización con diagnóstico de observacion por sepsis desde diciembre del 2013 hasta junio 2014. Se les realizó reacción en cadena de Polimerasa (PCR) de suero, mucosas y líquido cefalorraquídeo, las cuales fueron enviadas Departamento de Virología del Instituto Conmemorativo Gorgas de estudios de la Salud (ICGES) para ser procesadas por Enterovirus, virus Herpes y Citomegalovirus. Resultados: Ingresaron al Hospital del Niño un total de 345 neonatos con diagnóstico de Observación por sepsis en el periodo Dic 2013 a Junio 2014. Cumplieron los criterios de inclusión a nuestro estudio 60 neonatos. De estos un 15 % de casos de infección por enterovirus, 2% de casos de infección por citomegalovirus y ningún caso de infección por virus Herpes. Conclusión: El 4.7% de los neonatos nacidos en el HST son admitidos por observación por sepsis en nuestra institución. La frecuencia de infecciones virales en neonato febriles con sepsis clinica fue de 17%. El realizar de rutina la prueba de PCR viral en neonatos con sepsis clínica no solo permite el manejo adecuado y oportuno de estos casos sino que además podría representar menos gastos hospitalarios (menor estancia intrahospitalaria) y uso racional de antibióticos.

Introduction: Fever in infants is commonly associated with viral infections. Infection with herpes simplex virus (HSV) in neonates is associated with high morbidity and mortality and signi cant long-term sequelae. In recent years this infection has been an increase in incidence. The Enterovirus, is a well-known fever and cause aseptic meningitis in neonates. Cytomegalovirus infection in newborns is the most common congenital viral infection, a ecting 0.2% to 2.5% of all live births. The PCR test has been successfully used to identify CMV DNA, Herpes virus and Enterovirus in newborns. Materials and methods: All febrile neonate admission in Neonatal or another ward with a diagnosis of sepsis observation from December 2013 to June 2014. Patients underwent polymerase chain reaction (PCR) of serum, mucous evaluated and cerebrospinal fluid, which were sent Department of Virology of the Gorgas Memorial Institute for Health Studies (ICGES) to be processed by Enterovirus, Herpes virus and Cytomegalovirus. Results: Children admitted to hospital a total of 345 infants with sepsis diagnosis. Observing the period December 2013 to June 2014 met the criteria inclusion our study 60 infants. Of these 15% of cases of enterovirus infection, 2% of cases of cytomegalovirus infection and no cases of Herpes virus infection. Conclusion: 4.7% of infants born in the Hospital Santo Tomas are admitted for neonatal sepsis in our institution. The frequency of viral infections in febrile neonates with clinical sepsis was 17%. Performing routine viral PCR test in neonates with clinical sepsis not only allows adequate and timely handling of these cases but could also represent less hospital expenditures (less hospital stay) and rational use of antibiotics.

Variables clínicas asociadas al diagnóstico de meningo-encefalitis herpética en pacientes adultos: un estudio anidado de casos y controles / Clinical characteristics of adult patients with herpetic meningoencephalitis: a nested case control study

Introduction: Herpes simplex virus (HSV) is the most common etiology of sporadic encephalitis and presents with an estimated mortality of 50-70%. Objective: To describe baseline characteristics of patients with herpetic encephalitis admitted to a tertiary teaching hospital and their difference with patients with non herpetic encephalitis. Materials and Methods: Nested case control study using a retrospective cohort of patients with suspected encephalitis admitted to the Hospital Italiano de Buenos Aires (2006-2013). Adult patients included had a lumbar puncture with a positive or negative polimerase reaction for HSV. A case of herpetic encephalitis was defined as a positive polimerase reaction in spinal fluid. For each case, 5 controls were randomly selected. Results: There were no baseline differences present between cases and controls. The only covariate associated with herpetic encephalitis was an abnormal brain magnetic resonance imaging (MRI) (OR: 5.37, IC 95% 1.42-20.38, p < 0.01). The most frecuent alterations in the MRI were extratemporal lesions or temporal ones with or without haemorrhage. Discussion: There are no apparent baseline clinical differences between patients with or without herpetic encephalitis. A positive finding in a brain MRI should be taken into account during clinical workup.

Introducción: El virus herpes simplex (VHS) es la causa reportada más común de encefalitis esporádica con una mortalidad estimada de 50 a 70%. Objetivo: Describir las características de los pacientes con encefalitis herpética (EH) en nuestro medio y sus variables clínicas asociadas. Materiales y Métodos: Estudio anidado de casos y controles sobre una cohorte retrospectiva en el Hospital Italiano de Buenos Aires (2006-2013). Se incluyeron pacientes adultos con sospecha de encefalitis a quienes se les realizó una punción lumbar con posterior reacción de polimerasa en cadena para VHS en líquido cefalorraquídeo. Por cada caso (reacción positiva para VHS) se tomaron cinco controles aleatoriamente seleccionados. Resultados: No se observaron diferencias significativas en las características demográficas y clínicas entre los casos de EH y los controles. La única variable clínica asociada al diagnóstico de EH fue la alteración en la resonancia magnética (RM) de cerebro (OR: 5,37, IC 95% 1,4220,38; p < 0,01). Los patrones más comunes de alteración en la RM fueron las lesiones extra-temporales o lesiones temporales con o sin hemorragia. Discusión: Este hallazgo nos hace jerarquizar el hallazgo de un resultado positivo en la RM durante la valoración inicial de un paciente con clínica compatible de EH.

Infecciones de transmisión sexual silentes: la muestra de orina permite una adecuada detección / Non symptomatic sexually transmitted infections: urine sample allows an adequate detection

Detection of non-symptomatic sexually transmitted infections (NSSTD) has taken great relevance, primarily due to global increase. This has led to implement various laboratory techniques with the aim of early detection of these silent infections to decrease the incidence. Techniques usually used for the detection and identification of NSSTD require invasive samples (blood, citobrush, etc.), so the urine could be a simpler option and noninvasive sample when the patient be subjected to test for some of these infections.

La detección de infecciones de transmisión sexual silentes (ITSS) ha tomado gran relevancia, debido principalmente a su incremento en el mundo. Esto ha llevado a implementar diversas técnicas de laboratorio con la finalidad de la detección precoz de estas infecciones silentes para disminuir su incidencia. Las técnicas que habitualmente se utilizan para la detección e identificación de ITSS requieren de una muestra invasora (sangre, citobrush, entre otras), por lo que la orina podria ser una opción de muestra más simple y no invasora al momento que el paciente se deba someter a un examen para detectar alguna de estas infecciones.

Role of IL-4 in an experimental model of encephalitis induced by intracranial inoculation of herpes simplex virus-1 (HSV-1) / Papel da IL-4 em modelo experimental de encefalite induzida pela inoculação intracraniana do herpes simplex vírus-1 (HSV-1)

Herpes simplex virus-1 (HSV-1) is a pathogen that may cause severe encephalitis in humans. In this study, we aimed to investigate the role of interleukin-4 (IL-4) in a model of HSV-1 brain infection. IL-4 knockout (IL-4-/-) and wild type (WT) C57BL/6 mice were inoculated with 10(4) plaque-forming units of HSV-1 by the intracranial route. Histopathologic analysis revealed a distinct profile of infiltrating cells at 3 days post-infection (dpi). Infected WT mice presented mononuclear inflammatory cells while IL-4-/- mice developed meningoencephalitis with predominance of neutrophils. IL-4-/- mice had diminished leukocyte adhesion at 3 dpi when compared to infected WT animals in intravital microscopy study. Conversely no differences were found in cerebral levels of CXCL1, CXCL9, CCL3, CCL5 and TNF-α between WT and IL-4-/- infected mice. IL-4 may play a role in the recruitment of cells into central nervous system in this acute model of severe encephalitis caused by HSV-1.

O vírus herpes simplex-1 (HSV-1) é um patógeno que pode causar encefalite grave em humanos. Neste estudo, buscamos investigar o papel da interleucina-4 (IL-4) no modelo de infecção intracerebral por HSV-1. Camundongos C57BL/6 selvagens (WT) e deficientes no gene IL-4 (IL-4-/-) foram inoculados com 10(4) unidades formadoras de placas de HSV-1 por via intracraniana. A análise histopatológica revelou um padrão distinto de infiltrado leucocitário. Camundongos WT infectados apresentaram infiltrado de células mononucleares, enquanto camundongos IL-4-/- desenvolveram meningoencefalite com predomínio de neutrófilos 3 dias pós-infecção (dpi). Animais IL-4-/- tiveram menor adesão de leucócitos 3 dpi quando comparados aos animais WT infectados à microscopia intravital. Em contrapartida, não foram encontradas diferenças nos níveis cerebrais de CXCL1, CXCL9, CCL3, CCL5 e TNF-α entre camundongos WT e IL-4-/- infectados. Esses resultados sugerem que IL-4 pode desempenhar um papel no recrutamento de células no sistema nervoso central neste modelo agudo de encefalite grave causada pelo HSV-1.

Diagnóstico microbiológico de infecciones de transmisión Sexual: Parte II. ITS virales / Microbiologic diagnosis of sexually transmitted diseases: Part II. STD of viral origin

Sexually transmitted diseases are highly prevalent and a public health concern worldwide. Herpes simplex virus (HSV) and human papilloma virus (HPV) are described. The development of rapid, sensible and specific diagnostic assays has been difficult because of their pathogenic features. In the last years, molecular biology based techniques allowed a better and wider range of diagnosis, as in the HPV-cancer association. In this article, standardized diagnostic methodologies for HSV and HPV are reviewed.

Las Infecciones virales de transmisión sexual son altamente prevalentes y constituyen un problema de salud pública en el mundo. Entre los agentes que se contagian por esta vía, se describe acerca de virus herpes simplex (HSV) y virus papiloma humano (HPV). Las características patogénicas de estas infecciones han dificultado la implementación de técnicas diagnósticas rápidas, sensibles y específicas para el diagnóstico clínico habitual. En los últimos años las metodologías diagnósticas sustentadas en la biología molecular han permitido mej orar y ampliar el rango de diagnóstico posible para estos agentes infecciosos y relacionarlos con otras patologías, como es el caso de HPV y cáncer. En el presente artículo se revisan metodologías diagnósticas implementadas para el diagnóstico microbiológico de HSV y de HPV.

Eritema multiforme ampollar extenso asociado a infección por virus herpes simplex / Extended Bullous Erythema Multiforme Associated To Herpes Simplex Virus Infection

El Eritema Multiforme (EM) es una reacción cutánea aguda generalmente benigna y autolimitada, asociada a la infección por Virus Herpes Simplex (HSV). Se caracteriza por lesiones polimorfas y tipo diana en extremidades y mucosas. Presentamos un paciente de 22 años con pápulas, vesículas y ampollas, que evoluciona con un 90% de la superficie corporal comprometida en tres semanas. Se realizó una reacción de polimerasa en cadena para HSV, resultando positiva en una costra. La biopsia de piel y la tinción de inmunohistoquímica positiva para linfocitos T CD4, fueron compatibles con EM ampollar asociado a HSV. Destacamos la importancia de la correlación clínico patológica, apoyada por el estudio virológico, en el diagnóstico de este caso de presentación atípica. Los hallazgos de laboratorio confirmaron lo descrito en la literatura respecto de la patogenia del EM asociado a HSV.

Erythema Multiforme (EM) is a generally benign and self-limited acute cutaneous reaction, associated with Herpes Simplex Virus (HSV) infection. It is characterized by polymorphic "target" lesions in extremities and mucosal tissues. We report a 22-year old patient with papules, vesicles and blisters, which evolved to cover 90% of the body in three weeks. We performed a PCR study for HSV, which was positive in a crust. A skin biopsy and positive immunohistochemical stain for LT CD4+ were compatible with bullous EM associated with HSV. We underline the importance of pathological clinical correlation, reinforced by a virological study, in the diagnosis of this case with atypical symptoms. The laboratory findings confirmed literature descriptions with respect to the pathogenicity of EM associated with HSV.

Eccema herpético por virus herpes simplex tipo 2: revisión de la literatura a propósito de un caso / Eczema herpeticum caused by herpes simplex virus type 2: review of the literature about one case

Eczema herpeticum is an extensive vesicular eruption caused by cutaneous dissemination of herpes simplex virus in patients who have underlying skin conditions or epidermal barrier disruption. Herpes simplex virus type 1 (HSV 1) is the most common etiology. This is the first report in Chile of a case of eczema herpeticum caused by HSV 2 in a patient with Darier' s disease, We review the literature and emphasize the use of sensitive and specific assays as real time polimerase chain reaction, that allow to initiate timely the correct treatment.

El eccema herpético es una erupción vesicular extensa causada por la diseminación cutánea del virus herpes simplex, en pacientes con trastornos cutáneos previos o daño de la barrera epidérmica. La etiología más frecuente es virus herpes simplex tipo 1 (VHS 1), aunque se han reportado casos de virus herpes simplex tipo 2 (VHS 2). Presentamos el primer reporte en Chile de eccema herpético por VHS 2 en un paciente con enfermedad de Darier. Realizamos una revisión de la literatura médica y destacamos la importancia del uso de técnicas sensibles y específicas como la reacción de polimerasa en cadena (RPC) en tiempo real, que posibilitan el inicio precoz de la terapéutica correcta.

Prevalência e fatores de risco para a infecção pelo herpes simples vírus tipo-2 em uma coorte de homens que fazem sexo com homens no Rio de Janeiro / Prevalence and risk factors for herpes simplex type-2 virus in a cohort of men who have sex with men in Rio de Janeiro

O HSV-2 é a principal causa de herpes genital, sendo a maior parte destas infecções assintomática. A infecção pelo HSV-2 pode ser um fator de risco para a aquisição e transmissão de outras DST, inclusive da infecção causada pelo HIV. Objetivo: Estimar a prevalência e identificar os fatores de risco associados à infecção pelo HSV-2 em uma coorte de homens que fazem sexo com homens da Fundação Oswaldo Cruz, Rio de Janeiro, Brasil. Métodos: Estudo transversal, com avaliação de dados sócio-demográficos, comportamentais e laboratoriais referentes à linha de base de 403 voluntários incluídos na referida coorte, no período de 1994-1998. As amostras foram testadas através do ELISA para detecção de anticorpos tipo-específicos contra a gG2 do HSV-2 (HerpeSelect - Focus Technologies)Resultados: A prevalência para anticorpos anti-HSV-2 encontrada nesta população foi de 65,2 por cento. Após análise multivariada, os fatores que se mostraram independentemente associados à infecção pelo HSV-2 foram idade mais elevada (maior ou igual a 26 anos), raça não branca, ter sorologia positiva para sífilis (TPHA), ter feito sexo desprotegido com parceria sexual masculina e ter feito sexo desprotegido com parceria sexual feminina nos seis meses que antecederam a visita. Conclusão: Este estudo mostra uma elevada prevalência da infecção pelo HSV-2 neste grupo e associação dessa infecção com outras DST, em concordância com outros estudos. Estes dados podem ter uma importante contribuição no sentido de avaliar estratégias que possam contribuir para a prevenção desta infecção na população de homens que fazem sexo com homens, que é tida como uma população vulnerável e central à dinâmica da disseminação do HSV-2 em todo o mundo...

HSV-2 is the primary cause of genital herpes and most of the infections are asymptomatic. The infection by HSV-2 may be a risk factor to acquisition and transmission of other sexually transmitted diseases (STD), including infection by HIV virus. Objective: Estimate the prevalence and identify the risk factors associated of infection by HSV-2 in a cohort of men who have sex with men (MSM) in Fundação Oswaldo Cruz [Oswaldo Cruz Foundation], Rio de Janeiro, Brazil.Methods: Cross-sectional study, evaluating sociodemographic, behavioral, and laboratorialdata from baseline of 403 volunteers included in this cohort, in the period of 1994-1998. The samples were tested by ELISA to detect type-specific antibodies against HSV-2 gG2 (HerpeSelect - Focus Technologies). Results: The prevalence to antibodies anti-HV2-2 observed in this population was 65.2 percent.After multivariate analysis, the factors independently associated to HSV-2 infection were older age (> 26 years), non-white race, positive serology for syphilis (TPHA), history of unprotected sex with male partners and history of unprotected sex with female partners inthe last six months before the visit.Conclusion: This study shows a high prevalence of HSV-2 infection in this group and association with other STD, consistent with other studies. This data may have a significant contribution to evaluate strategies that may contribute to prevent this infection amongMSM, as this is considered a vulnerable core-population to the dynamics of HSV-2 spreading all over the world...

In vitro antiviral activity of Chamaecrista nictitans (Fabaceae) against herpes simplex virus: biological characterization of mechanisms of action

We have previously identified a crude extract of the plant Chamaecrista nictitans (Fabaceae) with antiviral activity against herpes simplex virus. The main objectives of this research were to identify the step of the replication cycle of herpes simplex inhibited by the extract, and to attempt to characterize the chemical characteristics of this extract. The crude extract from--Chamaecrista nictitans (Fabaceae) was extracted with a mixture of diclorometane/methanol, and further fractionated following a bioassay-guided protocol using a combination of preparative thin layer and column chromatography. Toxicity and bioassay experiments were carried out in monolayers of Vero cells. The antiviral activity of the extract was assessed by total inhibition of cytopathic effect after three-day incubation. The highest concentration of the extract which was not toxic to the cells was 200 ptg/ml. Western blot and immunofluorescence techniques were used to elucidate the antiviral mechanism of the extract by infecting Vero cells with the virus at different times and monitoring the synthesis of viral proteins. A 60 kDa protein was detected at 2 hr and 8 hr post-infection but no additional proteins were synthesized at later time intervals, and cytopathic effect was not observed after 24 hr. This result indicates that the extract acts at the intracellular level in order to inhibit late transcription. However, it does not inhibit transcription/translation of early viral proteins. These results were confirmed by immunofluorescence experiments. A strong fluorescent signal was observed in control cell monolayers at 24 hr post infection, accompanied with a clear cytopathic effect. In contrast, in the presence of acyclovir or the extract, cells showed very discrete immunofluorescence, characterized by a punctuated pattern, and no cytopathic effect was observed. Neutralization assays were performed using pre-incubation of virus with either specific herpes simplex-1 antiserum, 200...