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We report the histopathological study of a large, black, crusted lesion with symmetrical distribution in both buttocks and perineum, never described, in a man who has sex with men (MSM) and proctitis associated with Human Monkey Pox Virus (hMPXV) and HIV-AIDS infection never treated. A 39-year-old male, homosexual, HIV-AIDS without Highly Active Antiretroviral Therapy (HAART), was admitted to a hospital in Lima, Peru, with papulopustular lesions on the body and perianal area. Days later, a large, crusty, black lesion with a symmetrical distribution appeared on the buttocks and perineum. The tissue culture was negative. Wedge biopsy of the lesion showed typical MPXV cytopathogenics lesions in addition to fibrin micro thrombosis in the underlying papillary dermis. The histopathological findings of the scabby and black lesion are the classic ones described by Stagles, except for the phenomenon of fibrin micro thrombosis in the papillary dermis, a novel cytopathogenic effect of MPXV with clinical relevance (epidermal-dermal necrosis).
Reportamos el estudio histopatológico de una gran lesión costrosa, negra, de distribución simétrica en ambas nalgas y periné, nunca descrito, en un hombre que tiene sexo con hombres (HSH) y proctitis asociado a Viruela del Mono Humana (hMPXV) e infección por VIH-SIDA nunca tratado. Un varón de 39 años, homosexual, VIH-SIDA sin Terapia Antirretroviral de Gran Actividad (TARGA), ingresó en un hospital de Lima, Perú, con lesiones papulopustulosas en el cuerpo y el area perianal. Días después apareció una gran lesión negra, costrosa de distribución simétrica en las nalgas y periné. El cultivo de tejidos fue negativo. La biopsia en cuña de la lesión mostró lesiones citopatogénicas típicas de MPXV, además de microtrombosis de fibrina en la dermis papilar subyacente. Los hallazgos histopatológicos de la lesión costrosa y negra que reportamos son los clásicos descritos por Stagles, a excepción del fenómeno de microtrombosis de fibrina en la dermis papilar, un efecto citopatogénico novedoso de MPXV con relevancia clínica (necrosis epidermo-dérmico).
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Objective To investigate the effect of Andrographis paniculata Nees(APN)extract on Coxsackievirus A16(CVA16)in vitro.Methods African green monkey kidney-derived Vero cells(Vero cells)were treated with APN extract at the concentration of 500.0,250.0,125.0,60.0,30.0,15.0,7.5 and 3.8 μg/mL,the cytotoxicity was determined with cell counting Kit-8 and the IC50was calculated by Probit unit regression method.Direct inactivating activity on CVA16,blocking of CVA16 adsorbing Vero cells and inhibition of CVA16 replication in Vero cells were determined and compared between Ribavirin(RBV) and APN extract with CVA16-infected Vero cells.SPSS 24.0 software was used to analyze the data.Results The selected concentrations of APN extract and RBV for experiment were 15.0, 7.5 and 3.8 μg/mL according to cytotoxicity test.Both of APN extract and RBV had neither direct inactivation on CVA 16 nor blocking of CVA16 adsorbing at the concentration of 15.0,7.5 and 3.8 μg/mL(F=1.54,1.52 and 0.67, 1.68,all P>0.05).However,both drugs had the capability of inhibiting CVA 16 replication in Vero cells at the concentration of 15.0 and 7.5 μg/mL(t=6.87,11.76 and 7.71,12.84,all P<0.05).Conclusion Experimental result shows that APN extract can effectively inhibit CVA 16 replication in Vero cells in vitro.
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Introducción. Los virus entéricos se han visto implicados en brotes de enfermedad diarreica aguda, enfermedades transmitidas por alimentos, hepatitis A y meningitis aséptica, en los que el vehículo de transmisión del agente ha sido el agua.Objetivo. Estandarizar un método de concentración para la detección de virus entéricos en aguas de consumo.Materiales y métodos. Se concentraron 20 litros de agua a un volumen de 6 ml mediante filtración y ultrafiltración tangencial. Como controles positivos se prepararon soluciones de 20 litros a concentraciones virales de 1, 10, 50 y 100 TCID50 (Tissue Culture Infectious Dose 50%) de Poliovirus Sabin de tipo 1. Las partículas virales fueron recuperadas por cultivo en células sensibles a la infección e identificadas por amplificación del genoma viral mediante reacción en cadena de la polimerasa, siguiendo los estándares internacionales de los Centers for Disease Control and Prevention (CDC) de Atlanta. Resultados. Todos los controles positivos causaron efecto citopático en células de rabdomiosarcoma y L20B y fueron detectados por RT- PCR (Real Time- PCR) convencional, directamente de las muestras. Los controles negativos no mostraron efecto citopático ni amplificación viral por RT-PCR.Conclusiones. La ultrafiltración tangencial mostró ser un método rápido y eficaz al recuperar virus desde una TCID50, además de ser reproducible y sencillo. Tiene la ventaja de permitir la detección de su capacidad de contagiosidad viral por el cultivo celular, y la identificación por RT-PCR.
Introduction. Enteric viruses have been implicated in acute diarrheal disease, food-borne disease, hepatitis A and meningitis outbreaks, in which water was the vehicle of transmission.Objective. A concentration method was standardized for the detection of enteric viruses in drinking water. Materials and methods. Twenty liters of water were concentrated to 6 ml by filtration and tangential ultrafiltration. Viral solutions of 20 L each were prepared at 1, 10, 50 and 100 TCID50 of Sabin poliovirus type 1 as positive controls. Viral particles were recovered by tissue culture and detected by conventional polymerase chain reaction (PCR), according to the international standards recommended by the Enterovirus Laboratory at the Centers for Disease Control and Prevention, Atlanta, GA. Results. All positive controls showed cytopathic effect on L20B and RD cells and were amplified by conventional PCR directly from samples. Negative controls did not show any amplification or viral cytopathic effect. Conclusions. Tangential ultrafiltration for concentrating viruses proved to be a fast, efficient recovery and reproducible. It has the advantage of allowing the detection (at the 1 TCID50 level) and identification of viruses by RT-PCR and the demonstration of viral infectivity by tissue culture.
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Cytopathogenic Effect, Viral , Enterovirus , Poliovirus , Reverse Transcriptase Polymerase Chain Reaction , Water QualityABSTRACT
Objective To understand the sensitivity of cytopathogenic effect (CPE) in MadinDarby canine kidney cells(MDCK) that cultured influenza A pharyngeal swab specimens of patients for one,two and three passages. Methods Influenza A pharyngeal swab specimens of patients were inoculated in MDCK for three blind passages. The presence of CPE of every passage was observed by inverted microscope. Results Of the 279 influenza A pharyngeal swab specimens of patients tested by colloidal gold, the presence of CPE in MDCK for one,two and three passages was 65.9%(184/279),91.4%(255/279) and 96.4%(269/279), respectively. Two hundred and seventy-one of 279specimens were identified as influenza A by multiplex reverse transcription-polymerase chain reaction (RT-PCR). Conclusion The positive separation rate can reach more than 95% by inoculating influenza A pharyngeal swab specimens of patients in MDCK for three blind passages.
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AIM: To explore the relationship between human cytomegalovirus(HCMV) gene activity and cytopathogenesis in HCMV infection. METHODS: HCMV echelin-infected cell model was set up in vitro by coincubating passage cultured HEL and HCMV with different titers (group A: 10-5; group B: 10-3; and group C: 10-1). FQ-PCR was performed to evaluate the number of HCMV DNA copies. MCP mRNA was measured by RT-PCR. Meanwhile the development of cytopathologic effects (CPE) was observed under microscope and ultrastructural changes determined by TEM.RESULTS: Compared with group B and C in high HCMV titer, group A with low viral titer showed low HCMV DNA load (P