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1.
Braz. J. Pharm. Sci. (Online) ; 58: e18637, 2022. tab, graf
Article in English | LILACS | ID: biblio-1364416

ABSTRACT

Abstract The therapeutic drugs to treat Herpes simplex virus (HSV) infections have toxic side effects and there has been an emergence of drug-resistant strains. Therefore, the search for new treatments for HSV infections is mounting. In the present study, semi-solid formulations containing a crude hydroethanolic extract (CHE) from Schinus terebinthifolia were developed. Skin irritation, cutaneous permeation, and in vivo therapeutic efficacy of the formulations were investigated. Treatment with the ointment formulations did not result in any signs of skin irritation while the emulsions increased the thickness of the epidermis in Swiss mice. The cutaneous permeation test indicated that the CHE incorporated in the formulations permeated through the skin layers and was present in the epidermis and dermis even 3 h after topical application. In vivo antiviral activity in BALB/c mice treated with the CHE ointments was better than those treated with the CHE emulsions and did not significantly differ from an acyclovir-treated group. Taken together, this suggests that the incorporation of CHE in the ointment may be a potential candidate for the alternative topical treatment of herpetic lesions.


Subject(s)
Pharmaceutical Preparations/analysis , Simplexvirus/classification , Herpesvirus 1, Human/classification , Anacardiaceae/adverse effects , Antiviral Agents/adverse effects , Acyclovir/antagonists & inhibitors , Efficacy , Emulsions/adverse effects
2.
Dement. neuropsychol ; 15(2): 164-172, Apr.-June 2021. graf
Article in English | LILACS | ID: biblio-1286203

ABSTRACT

ABSTRACT. Considering the variety of mechanisms of Herpes simplex virus (HSV-1) contamination and its broad invasive potential of the nervous system, a life-long latent infection is established. Infected adult individuals may be susceptible to viral reactivation when under the influence of multiple stressors, especially regarding immunocompromised patients. This guides a series of neuroinflammatory events on the cerebral cortex, culminating, rarely, in encephalitis and cytotoxic / vasogenic brain edema. A sum of studies of such processes provides an explanation, even though not yet completely clarified, on how the clinical evolution to cognitive impairment and dementia might be enabled. In addition, it is of extreme importance to recognize the current dementia and cognitive deficit worldwide panorama. The aim of this literature review is to elucidate the available data upon the pathophysiology of HSV-1 infection as well as to describe the clinical panorama of the referred afflictions.


RESUMO. Considerando a variedade de mecanismos de contaminação pelo vírus Herpes simplex (HSV-1) e seu amplo potencial invasivo do sistema nervoso, uma infecção latente por toda a vida é estabelecida. Indivíduos adultos infectados podem ser suscetíveis à reativação viral quando estão sob a influência de múltiplos estressores, principalmente em pacientes imunocomprometidos. Esse fator orienta uma série de eventos neuroinflamatórios no córtex cerebral, culminando, raramente, em encefalite e edema cerebral citotóxico/vasogênico. Um somatório de estudos desses processos fornece uma explanação, embora ainda não totalmente esclarecida, de como a evolução clínica para déficit cognitivo e demência pode ser possibilitada. Além disso, é de extrema importância reconhecer o panorama mundial atual da demência e do déficit cognitivo. O objetivo da presente revisão de literatura é elucidar os dados disponíveis sobre a fisiopatologia da infecção pelo HSV-1, assim como descrever o panorama clínico das referidas afecções.


Subject(s)
Humans , Herpesvirus 1, Human , Central Nervous System Viral Diseases , Encephalitis, Herpes Simplex , Dementia , Cognitive Dysfunction
3.
Article in Chinese | WPRIM | ID: wpr-888082

ABSTRACT

In this study, emotional stress-induced herpes simplex virus type 1(HSV-1) susceptibility model was employed to simu-late the pathological state of " depression-induced liver fire", and the protection effect of Qingre Xiaoyanning(QX) in clearing liver fire was investigated. BALB/c mice were randomly divided into a normal group, a HSV-1 group, a restraint stress + HSV-1 group,low-(0. 658 g·kg~(-1)) and high-dose(1. 316 g·kg~(-1)) QX groups, and an acyclovir group. Except for the normal group and the HSV-1 group, the mice in other groups received daily restraint stress for 6 h from day 3 of medication. On day 9 of medication, mice were anesthetized by isoflurane and infected intranasally with HSV-1. Survival rate, weight change, encephalitis symptoms, and eye injury of mice were recorded for 14 d after virus infection. Hematoxylin-eosin(HE) staining and immunohistochemical staining were used to detect pathological changes and HSV-1 antigen distribution. Plaque assay was performed to detect the titer of HSV-1. The protein ex-pression of ICP27 in the mouse brain was detected by Western blot. The experimental results showed that QX could increase the survival rate of HSV-1-infected mice loaded with emotional stress(P<0. 001), reduce the titer of HSV-1 in the mouse brain(P<0. 01), relieve brain inflammation(P<0. 05) and eye injury(P<0. 05), down-regulate the expression of ICP27 related to HSV-1(P<0. 05), and decrease the distribution of HSV-1 antigen in the mouse brain. The results demonstrated that QX significantly reduced the susceptibility to HSV-1 induced by emotional stress, which is expected to provide a theoretical basis for the treatment and preven-tion of HSV-1 infection and promote the clinical development and application of Chinese medicine effective in clearing liver fire.


Subject(s)
Animals , Capsules , Herpes Simplex , Herpesvirus 1, Human , Mice , Mice, Inbred BALB C , Psychological Distress
4.
Rev. Hosp. Ital. B. Aires (2004) ; 40(4): 219-222, dic. 2020. ilus
Article in Spanish | LILACS | ID: biblio-1145550

ABSTRACT

La erupción variceliforme de Kaposi es una infección cutánea diseminada, causada en la mayor parte de los casos por el virus Herpes simple tipo 1. Se suele presentar en pacientes con alteraciones preexistentes de la barrera cutánea, especialmente en niños con dermatitis atópica. Se comunica el caso de un paciente de 84 años, quien negaba enfermedades cutáneas previas, que consultó por lesiones dolorosas y pruriginosas, en la piel del tórax y el abdomen, de 3 semanas de evolución. Con sospecha de una enfermedad infecciosa viral, bacteriana, ampollar o neutrofílica, se realizó inmunofluorescencia directa para herpes, cultivo y biopsia de piel para estudio histológico. La inmunofluorescencia fue positiva para Herpes simple tipo 1 y el estudio histopatológico mostró cambios compatibles con infección herpética y enfermedad de Darier. La enfermedad de Darier es una genodermatosis infrecuente que se suele manifestar en la adolescencia. Si bien su diagnóstico en la ancianidad es excepcional, este caso ilustra que se debe considerar en todos los pacientes que presenten erupción variceliforme. (AU)


Kaposi's varicelliform rash is a disseminated cutaneous infection, caused by Herpes virus 1. It usually presents in patients with pre-existing skin barrier disorders, especially in children with atopic dermatitis. We report the case of an 84-year-old patient, who reported having no previous skin diseases, who consulted for painful, itchy, 3-week-old skin lesions. As we suspected viral, bacterial, bullous or neutrophilic disease, direct immunofluorescence, culture, and skin biopsy for histological study were performed. Immunofluorescence was positive for Herpes simplex type 1 and the histopathological study showed changes compatible with herpetic infection and Darier's disease. Darier's disease is a rare genodermatosis that usually manifests in adolescence. Although its diagnosis in old age is anecdotal, it should be considered in patients with a varicelliform rash. (AU)


Subject(s)
Humans , Male , Aged, 80 and over , Kaposi Varicelliform Eruption/diagnosis , Darier Disease/diagnosis , Acyclovir/administration & dosage , Foscarnet/therapeutic use , Herpesvirus 1, Human/pathogenicity , Fluorescent Antibody Technique, Direct , Herpes Simplex/complications , Kaposi Varicelliform Eruption/etiology , Kaposi Varicelliform Eruption/pathology , Kaposi Varicelliform Eruption/drug therapy , Darier Disease/etiology
5.
Medisan ; 24(1)ene.-feb. 2020. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1091162

ABSTRACT

Introducción: Las enfermedades oftalmológicas originadas por virus ocupan un lugar importante en el quehacer diario de la especialidad. Objetivo: Caracterizar pacientes con infecciones oculares por herpesvirus según variables clínicas y epidemiológicas de interés. Métodos: Se realizó un estudio descriptivo y transversal de 29 pacientes adultos con infecciones por herpesvirus, atendidos en la consulta oftalmológica del Hospital General Docente Dr. Juan Bruno Zayas Alfonso de Santiago de Cuba, desde septiembre de 2012 hasta igual mes de 2013. Resultados: En la casuística predominaron los pacientes de piel blanca (55,1 %), el sexo femenino (69,0 %) y el grupo etario de 57 años y más (48,3 %). El ojo rojo y la inyección cilioconjuntival constituyeron los síntomas y signos más comunes (75,9 %). Las lesiones más frecuentes estuvieron localizadas en los párpados, la piel y la córnea (31,0 %). La afectación de la agudeza visual por debajo de 0,3 estuvo relacionada con la ocurrencia de afectación estromal y la localización de las lesiones en el área pupilar. Ningún paciente presentó inflamación del segmento posterior. Conclusiones: No se encontraron argumentos referidos a la relación existente entre la localización de las lesiones y la afectación de la agudeza visual, lo cual impidió realizar comparaciones.


Introduction: Eye diseases originated due to virus occupy an important place in daily routine of the specialty. Objective: To characterize patients with eye infections due to herpesvirus according to clinical and epidemiological variables of interest. Methods: A descriptive and cross-sectional study of 29 adult patients with infections due to herpesvirus was carried out, they were assisted in the ophthalmologic service of Dr. Juan Bruno Zayas Alfonso Teaching General Hospital in Santiago de Cuba, from September, 2012 to the same month in 2013. Results: In the case material white skin patients (55.1 %), female sex (69.0 %) and 57 years and over age group (48.3 %) prevailed. Red eye and ciliumconjuntival injection constituted the most common symptoms and signs (75.9 %). The most frequent lesions were located in the eyelids, the skin and cornea (31.0 %). The affectation of the visual acuteness below 0.3 was related to the occurrence of stromal affectation and the localization of lesions in the pupilar area. No patient presented inflammation of the later segment. Conclusions: Arguments about the existent relationship between the localization of lesions and the affectation of the visual acuteness were not found, which prevented to carry out comparisons.


Subject(s)
Eye Infections, Viral , Eye Infections, Viral/epidemiology , Herpesvirus 2, Human , Herpesvirus 1, Human , Eye Diseases
6.
Rev. bras. ginecol. obstet ; 42(1): 5-11, Jan. 2020. tab
Article in English | LILACS | ID: biblio-1092632

ABSTRACT

Abstract Objective Estimate the prevalence of human herpesvirus type 1 HSV-1 DNA in placental samples, its incidence in umbilical cord blood of newborns and the associated risk factors. Methods Placental biopsies and umbilical cord blood were analyzed, totaling 480 samples, from asymptomatic parturients and their newborns at a University Hospital. Nested polymerase chain reaction (PCR) and gene sequencingwere used to identify the virus; odds ratio (OR) and relative risk (RR) were performed to compare risk factors associated with this condition. Results The prevalence of HSV-1 DNA in placental samples was 37.5%, and the incidence in cord blood was 27.5%. Hematogenous transplacental route was identified in 61.4% from HSV-1+ samples of umbilical cord blood paired with the placental tissue. No evidence of the virus was observed in the remaining 38.6% of placental tissues, suggesting an ascendant infection from the genital tract, without replication in the placental tissue, resulting in intra-amniotic infection and vertical transmission, seen by the virus in the cord blood. The lack of condom use increased the risk of finding HSV-1 in the placenta and umbilical cord blood. Conclusion The occurrence of HSV-1 DNA in the placenta and in cord blood found suggests vertical transmission from asymptomatic pregnant women to the fetus.


Resumo Objetivo Estimar a prevalência do DNA do vírus herpes humano 1 (HSV-1) em amostras de placenta, sua incidência no sangue do cordão umbilical de recém-nascidos e fatores de risco associados. Métodos Biópsias de placenta e de sangue de cordão umbilical foram analisadas, totalizando 480 amostras de parturientes assintomáticas e seus recém-nascidos emum hospital universitário. Reação de cadeia de polimerase (RCP) nested e sequenciamento gênico foram usados para identificar o vírus; odds ratio (OR) e risco relativo (RR) foram realizados para comparar os fatores de risco associados à essa condição. Resultados A prevalência do DNA do HSV-1 em amostras de placenta foi de 37,5%, e a incidência no sangue do cordão foi de 27,5%. A via transplacentária hematogênica foi identificada em 61,4% das amostras de HSV-1+do sangue do cordão umbilical, pareadas com o tecido placentário. Nenhuma evidência do vírus foi observada nos restantes 38,6% dos tecidos placentários, sugerindo uma infecção ascendente do trato genital. A falta de uso do preservativo aumentou o risco de encontrar o HSV-1 na placenta e no sangue do cordão umbilical. Conclusão A ocorrência de DNA do HSV-1 na placenta e no sangue do cordão umbilical sugere uma transmissão vertical de gestantes assintomáticas para o feto.


Subject(s)
Humans , Female , Pregnancy , Infant, Newborn , Adult , Young Adult , Pregnancy Complications, Infectious/epidemiology , Herpesvirus 1, Human/isolation & purification , Herpes Simplex/epidemiology , Placenta/virology , Pregnancy Complications, Infectious/blood , Prenatal Care , Socioeconomic Factors , Brazil/epidemiology , DNA, Viral/analysis , Polymerase Chain Reaction , Incidence , Prevalence , Risk Factors , Infectious Disease Transmission, Vertical , Fetal Blood/virology , Herpes Simplex/blood , Herpes Simplex/transmission
7.
Article in Korean | WPRIM | ID: wpr-738588

ABSTRACT

PURPOSE: We report a case of herpes simplex keratitis after Descemet membrane endothelial keratoplasty (DMEK). CASE SUMMARY: A 67-year-old male underwent DMEK in his left eye due to pseudophakic bullous keratopathy. One week after DMEK, re-bubbling was performed due to partial detachment of Descemet's membrane at the corneal periphery. After re-bubbling, the cornea remained clear and the patient's visual acuity gradually improved. Two months after DMEK, the patient presented with mild discomfort and decreased visual acuity. The cornea showed an irregular, narrow dendrite with an epithelial defect and surrounding opacity. After confirming that Descemet's membrane was attached, the patient was started on oral valacyclovir for suspected herpes keratitis. Herpes simplex virus type 1 was eventually identified by polymerase chain reaction. The corneal lesion resolved after three weeks of antiviral treatment. CONCLUSIONS: Similar to penetrating keratoplasty, DMEK can trigger outbreaks of herpes simplex keratitis. Herpes simplex keratitis should remain on the clinician's differential diagnosis for patients who present with a corneal epithelial irregularity and decreased visual acuity following DMEK.


Subject(s)
Aged , Cornea , Corneal Transplantation , Dendrites , Descemet Membrane , Diagnosis, Differential , Disease Outbreaks , Herpes Simplex , Herpesvirus 1, Human , Humans , Keratitis , Keratitis, Herpetic , Keratoplasty, Penetrating , Male , Polymerase Chain Reaction , Visual Acuity
8.
Article in Chinese | WPRIM | ID: wpr-775249

ABSTRACT

Herpes simplex virus (HSV), including HSV-1 and HSV-2, is an important pathogen that can cause many diseases. Usually these diseases are recurrent and incurable. After lytic infection on the surface of peripheral mucosa, HSV can enter sensory neurons and establish latent infection during which viral replication ceases. Moreover, latent virus can re-enter the replication cycle by reactivation and return to peripheral tissues to start recurrent infection. This ability to escape host immune surveillance during latent infection and to spread during reactivation is a viral survival strategy and the fundamental reason why no drug can completely eradicate the virus at present. Although there are many studies on latency and reactivation of HSV, and much progress has been made, many specific mechanisms of the process remain obscure or even controversial due to the complexity of this process and the limitations of research models. This paper reviews the major results of research on HSV latency and reactivation, and discusses future research directions in this field.


Subject(s)
Herpes Simplex , Virology , Herpesvirus 1, Human , Physiology , Humans , Virus Activation , Physiology , Virus Latency , Physiology , Virus Replication
9.
Clinical Endoscopy ; : 606-611, 2019.
Article in English | WPRIM | ID: wpr-785663

ABSTRACT

Eosinophilic esophagitis is a rare disease in Asian countries, but its incidence is growing rapidly in Western countries. The main pathophysiology of eosinophilic esophagitis is esophageal epithelial barrier dysfunction; disruption of the esophageal epithelial barrier easily induces antigen sensitization to foods and aeroallergens, which leads to subsequent esophageal inflammation as a result of eosinophil recruitment. Here we report a case of an 11-year-old Korean boy who suffered from fever, odynophagia, dysphagia, and chest pain. His upper endoscopic findings showed longitudinal ulcers with a volcano-like appearance at the distal esophagus. Polymerase chain reaction test results and biopsy specimens were positive for herpes simplex virus type 1. He was treated with acyclovir and a proton pump inhibitor, but his follow-up endoscopy showed typical patterns of eosinophilic esophagitis, and the biopsy specimens were compatible with the diagnostic criteria for eosinophilic esophagitis. Therefore, we report a very rare case of eosinophilic esophagitis after herpes esophagitis in a Korean child with normal immunity.


Subject(s)
Acyclovir , Asian Continental Ancestry Group , Biopsy , Chest Pain , Child , Deglutition Disorders , Endoscopy , Eosinophilic Esophagitis , Eosinophils , Esophagitis , Esophagus , Fever , Follow-Up Studies , Herpesvirus 1, Human , Humans , Incidence , Inflammation , Male , Polymerase Chain Reaction , Proton Pumps , Rare Diseases , Simplexvirus , Ulcer
10.
Article in Chinese | WPRIM | ID: wpr-813038

ABSTRACT

To explore the antiviral activity of nano-realgar against herpes simplex virus Type II (HSV-2) in vitro.
 Methods: Acyclovir (ACV) as a positive control, the cytotoxicity of nano-realgar at different concentrations (including 200.00, 150.00, 100.00, 50.00, 25.00, 12.50, 6.25, 3.13, 1.54, 0.78, 0.39 and 0 mg/L) on normal Vero cells were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. HSV-2 virus titer was determined by plaque assay, and the Vero cells model of HSV-2 infection was established. Subsequently, the antiviral effects of nano-realgar at different concentrations (including 20.00, 10.00, 5.00, 2.50, 1.25, 0.63, 0.31, 0.15, 0.08, 0.04 and 0 mg/L) on infected cells model were evaluated by the observation of cytopathic effect (CPE) and MTT method under the 3 modes including pre-treatment, treatment and direct inactivation.
 Results: The 50% cytotoxic concentration (CC50) of nano-realgar on Vero cells was 37.15 mg/L. The titer of HSV-2 was 7.30 log PFUs/mL. In the 3 modes, the half-maximal effective concentration (EC50) of nano-realgar on HSV-2 infected Vero cells were 0.13, 1.80 and 0.52 mg/L, and the corresponding therapeutic index (TI) were 285.77, 20.64, 71.44, respectively. The TI value of nano-realgar on pre-treatment mode was higher than that of nano-realgar on treatment and direct inactivation modes.
 Conclusion: Nano-realgar can play a good anti-HSV-2 activity in the 3 modes (pre-treatment, treatment and direct inactivation), and the anti-HSV-2 efficacy of nano-realgar on pre-treatment mode is better than that of nano-realagr on other 2 modes.


Subject(s)
Animals , Antiviral Agents , Arsenicals , Chlorocebus aethiops , Herpesvirus 1, Human , Herpesvirus 2, Human , Sulfides , Vero Cells
11.
São Paulo; s.n; s.n; 2019. 88 p. graf.
Thesis in Portuguese | LILACS | ID: biblio-1015357

ABSTRACT

O câncer cervical é um dos tipos de câncer mais comuns entre as mulheres, e a infecção persistente pelos HPV-16 e HPV-18 é responsável por 70% dos casos. As vacinas profiláticas disponíveis possuem alta eficácia na prevenção da infecção pelos tipos mais prevalentes de HPV. No entanto, este tipo de abordagem não beneficia mulheres que já apresentam lesões precursoras ou tumores cervicais avançados, e a busca por abordagens terapêuticas para esse tipo de câncer é considerada uma necessidade. A qualidade do antígeno representa um aspecto fundamental para o sucesso de vacinas terapêuticas baseadas em proteínas recombinantes. Neste sentido, os sistemas de expressão em células eucarióticas, como leveduras e células de mamíferos são considerados adequados para a produção de proteínas com aplicação biotecnológica. O objetivo principal deste trabalho contemplou a expressão das proteínas de fusão gDE7E6 do HPV-16 e do HPV-18 e a oncoproteína E7 do HPV-16 em células da levedura Pichia pastoris e expressão da gDE7E6 do HPV-16 e do HPV-18 em células de mamífero HEK293T e CHODG-44 para obtenção de antígenos purificados com futura aplicação em vacinas terapêuticas contra tumores associados ao HPV-16 e HPV-18. Os genes que codificam as proteínas gDE7E6 dos HPV-16 e HPV-18 e da E7 do HPV-16 foram clonados no vetor pPIC9K, os quais foram linearizados por digestão enzimática e utilizados na transformação da P. pastoris. A expressão das proteínas foi analisada nos tempos de 24, 48, 72 e 96 horas, no entanto, não foi observada a produção das proteínas no sobrenadante e nem no lisado celular. Diante desta constatação, iniciamos a expressão das proteínas gDE7E6 do HPV-16 e gDE7E6 do HPV-18 em células de mamíferos HEK293T e CHODG-44. As sequências genéticas das proteínas gDE7E6 do HPV-16 e do HPV-18 foram clonadas no vetor de expressão pNU1 e analisadas por digestão enzimática. Análises de SDS-PAGE e western blot demonstraram a expressão das proteínas gDE7E6 do HPV-16 e do HPV-18 em até 96 horas em células HEK293T. Em paralelo, realizamos a transfecção estável dos plasmídeos contendo as sequencias da gDE7E6 do HPV-16 e gDE7E6 do HPV-18 em células CHO-DG44. Com o intuito de aumentar a expressão das proteínas de interesse na população mista de CHODG-44, realizamos amplificação genômica com metotrexato (MTX), sendo possível observar aumento da expressão das proteínas, conforme aumento gradativo nas concentrações de MTX. Posteriormente, foram feitas tentativas para isolar um clone produtor das proteínas gDE7E6 HPV-16 e HPV-18, através de clonagem por diluição limitante e sistema automatizado, sendo possível isolar um clone para cada construção através de matriz semisólida, confirmado por western blot e citometria de fluxo. Apesar de demonstrar a expressão das proteínas de interesse em sistema de expressão baseado em células de mamífero, o rendimento obtido após a purificação por afinidade ao níquel foi extremamente baixo, o que dificulta a obtenção dos antígenos para fins vacinais


Cervical cancer is one of the most common cancers among women, and persistent infection with HPV-16 and HPV-18 accounts for 70% of the cases. Available prophylactic vaccines are highly effective in preventing infection by the most prevalent types of HPV. However, this type of approach does not benefit women who already have precursor lesions or advanced cervical tumors, and the search for therapeutic approaches to this type of cancer is considered a necessity. Antigen quality represents a key aspect for the success of therapeutic vaccines based on recombinant proteins. In this sense, expression systems based in eukaryotic cells such as yeast and mammalian cells are considered suitable for the production of proteins with biotechnological applications. The main objective of this work was to express the gDE7E6 fusion proteins HPV-16 and HPV-18 and the E7 oncoprotein HPV-16 in Pichia pastoris and expression of gDE7E6 HPV-16 and HPV-18 in mammalian cells HEK293T and CHODG-44 to obtain purified antigens with future applications in therapeutic vaccines against HPV-16 and HPV-18 associated tumors. The genes encoding the gDE7E6 proteins HPV-16 and HPV-18 and E7 HPV-16 were cloned into the pPIC9K vector, which were linearized by enzymatic digestion and used in the transformation of P. pastoris. Expression of the proteins was analyzed at 24, 48, 72 and 96 hours, however, the production of the proteins in the supernatant and in the cell lysate was not observed. In light of this finding, we initiated the expression of gDE7E6 proteins HPV-16 and HPV-18 in mammalian cells HEK293T and CHODG-44. The genetic sequences of gDE7E6 proteins HPV-16 and HPV-18 were cloned into the pNU1 expression vector and analyzed by enzymatic digestion. SDSPAGE and western blot analyzes demonstrated expression of gDE7E6 proteins HPV-16 and HPV-18 within 96 hours in HEK293T cells. In parallel, we performed stable transfection of plasmids containing gDE7E6 HPV-16 and HPV-18 sequences into CHODG44 cells. In order to increase the expression of the proteins in the mixed population of CHODG-44, we performed genomic amplification with methotrexate (MTX), and it was possible to observe an increase in protein expression, as a gradual increase in MTX concentrations. Therefore, attempts were made to isolate a clone producing gDE7E6 proteins HPV-16 and HPV-18 by limiting dilution and automated system, being possible to isolate one clone for each construct through a semisolid matrix, confirmed by western blot and flow cytometry. Despite observing protein expression in mammalian cell-based expression system, the yield obtained after nickel affinity purification was extremely low, which makes it difficult to obtain the antigens for vaccine purposes


Subject(s)
Oncogene Proteins/classification , Human papillomavirus 16 , Human papillomavirus 18 , Pichia , Uterine Cervical Neoplasms/physiopathology , Herpesvirus 1, Human , Eukaryota , Antigens/analysis
13.
Arch. argent. pediatr ; 116(3): 468-470, jun. 2018. ilus
Article in Spanish | LILACS, BINACIS | ID: biblio-950029

ABSTRACT

En los neonatos, la parálisis facial es muy infrecuente y, por lo general, diagnosticada al nacer. Se presenta el primer caso de parálisis facial neonatal con identificación del virus del herpes simple 1 en el líquido cefalorraquídeo. Un varón de 35 días de vida acudió a Urgencias por la desviación de la comisura bucal hacia la izquierda y la ausencia de cierre del ojo derecho, sin sintomatología infecciosa ni antecedentes relevantes. La exploración física fue compatible con parálisis facial periférica. Las exploraciones complementarias de urgencia (hemograma, bioquímica, coagulación y citoquímica de líquido cefalorraquídeo) fueron normales. Fue ingresado con prednisolona oral y aciclovir intravenoso. La resonancia magnética craneal fue normal. A las 48 horas, se recibió el resultado positivo de la reacción en cadena de la polimerasa para el virus del herpes simple 1 en el líquido cefalorraquídeo. Con evolución favorable, completó 7 días de prednisolona oral y fue dado de alta tras 21 días de aciclovir intravenoso, con exploración neurológica previa normal.


Neonatal facial palsy is very uncommon and is generally diagnosed at birth. We present the first published case of neonatal facial palsy with identification of herpes simplex virus 1 in cerebrospinal fluid. A 35-day-old male was presented at the Emergency Department with mouth deviation to the left and impossibility of full closure of the right eye. There were no symptoms of infection or relevant medical history. Physical examination was compatible with peripheral facial palsy. Studies performed at admission were normal (blood count, biochemical analysis and coagulation blood tests and cerebrospinal fluid analysis). The patient was admitted on oral prednisolone and intravenous aciclovir. Cranial magnetic resonance was normal. Polymerase chain reaction test for herpes simplex virus 1 in cerebrospinal fluid was reported positive after 48 hours of admission. Patient followed good evolution and received prednisolone for 7 days and acyclovir for 21 days. At discharge, neurological examination was normal.


Subject(s)
Humans , Male , Infant , Herpesvirus 1, Human/isolation & purification , Facial Paralysis/diagnosis , Herpes Simplex/diagnosis , Antiviral Agents/administration & dosage , Acyclovir/administration & dosage , Prednisolone/administration & dosage , Cerebrospinal Fluid/virology , Treatment Outcome , Facial Paralysis/drug therapy , Facial Paralysis/virology , Glucocorticoids/administration & dosage , Herpes Simplex/drug therapy
14.
Article in English | WPRIM | ID: wpr-717056

ABSTRACT

BACKGROUND: Human herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) are responsible for a plethora of human diseases, of which cutaneous and mucocutaneous infections are the most prevalent. In its most severe form, HSV infection can cause meningitis/encephalitis. We compared the Luminex ARIES HSV 1&2 assay (Luminex Corp., Austin, TX, USA), an automated sample-to-result molecular solution, to two non-automated HSV DNA assays. METHODS: A total of 116 artificial controls were used to determine the analytical performance of the ARIES assay. Controls were prepared by spiking universal transport medium (UTM) and cerebrospinal fluid (CSF) samples from patients who tested negative for HSV by an in-house HSV-1 and -2 DNA assay with reference materials (SeraCare Life Sciences, MA, USA; ZeptoMetrix Corp., MA, USA). Another 117 clinical samples were then used to compare the clinical performance of the ARIES assay with those of an in-house assay and the FTD Neuro 9 assay (Fast Track Diagnostics, Junglinster, Luxembourg). RESULTS: The analytical sensitivity (95% limit of detection) of the ARIES assay was 318 copies/mL (UTM samples) and 935 copies/mL (CSF samples) for HSV-1 strain 96 and 253 copies/mL (UTM samples) and 821 copies/mL (CSF samples) for HSV-2 strain 09. No cross-reactivity was observed in samples spiked with 14 non-HSV microorganisms. Compared with the reference result (agreement between the in-house and FTD Neuro 9 results), the ARIES assay had overall concordance rates of 98.2% (111/113) and 100% (113/113) for HSV-1 and HSV-2, respectively. CONCLUSIONS: The ARIES assay appears to be an excellent alternative for rapid detection and differentiation of HSV in skin and genital infections, meningitis, and encephalitis.


Subject(s)
Biological Science Disciplines , Cerebrospinal Fluid , DNA , Encephalitis , Herpes Simplex , Herpesvirus 1, Human , Herpesvirus 2, Human , Humans , Meningitis , Real-Time Polymerase Chain Reaction , Simplexvirus , Skin
15.
Arq. bras. oftalmol ; 80(2): 84-87, Mar.-Apr. 2017. tab
Article in English | LILACS | ID: biblio-838786

ABSTRACT

ABSTRACT Objective: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. Methods: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. Results: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. Conclusions: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection.


RESUMO Objetivo: Ceratites bacterianas ocorrem mundialmente e apesar dos novos desenvolvimentos permanece como uma condição que pode levar à cegueira. Avaliar a presença de herpes simples (-1 e -2) e vírus varicella zoster (VZV) por reação em cadeia quantitativa de polimerase em tempo real (qPCR) em raspados corneanos de pacientes com ceratite bacteriana. Métodos: Sessenta e cinco pacientes com ceratite infecciosa foram submetidos a raspados corneanos estudados para gram, Giemsa, cultura e qPCR (grupo de estudo). Foram avaliados fatores de risco e epidemiológicos. O grupo controle foi composto por 25 casos de úlcera dendrítica típica por herpes analisados por qPCR. Resultados: Do grupo de estudo (n=65), nove pacientes (13,8%) apresentaram cultura, qPCR e raspado negativos. Cinquenta e seis (86,2%) pacientes apresentaram cultura positiva, 51 para bacteria, 4 para fungo e 1 para ameba. A qPCR identificou 10 pacientes do grupo de cultura positiva para bactéria que também foram positivos para vírus, um VZV e 9 para HSV-1. Dos 25 pacientes que compunham o grupo controle, 21 apresentaram qPCR positivo para HSV-1. Conclusão: Herpes pode estar presente em pacientes com úlceras de córnea bacterianas e a qPCR pode ser útil na sua detecção.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Keratitis, Dendritic/microbiology , Herpesvirus 2, Human/isolation & purification , Herpesvirus 1, Human/isolation & purification , Herpesvirus 3, Human/isolation & purification , Cornea/virology , Real-Time Polymerase Chain Reaction/methods , Keratitis/microbiology , DNA Probes , Eye Infections, Bacterial/microbiology , Keratitis, Dendritic/diagnosis , Keratitis, Dendritic/virology , Prospective Studies , Keratitis/diagnosis , Keratitis/virology
16.
Mem. Inst. Oswaldo Cruz ; 112(3): 220-223, Mar. 2017. tab, graf
Article in English | LILACS | ID: biblio-841773

ABSTRACT

The use of quantitative real time polymerase chain reaction (qPCR) for herpesvirus detection has improved the sensitivity and specificity of diagnosis, as it is able to detect shedding episodes in the absence of clinical lesions and diagnose clinical specimens that have low viral loads. With an aim to improve the detection and quantification of herpesvirus by qPCR, synthetic standard curves for human herpesvirus 1 and 2 (HHV-1 and HHV-2) targeting regions gD and gG, respectively, were designed and evaluated. The results show that synthetic curves can replace DNA standard curves in diagnostic herpes qPCR.


Subject(s)
Humans , Herpesvirus 2, Human/genetics , Herpesvirus 1, Human/genetics , Herpes Simplex/virology , DNA, Viral/genetics , Sensitivity and Specificity , Viral Load , Real-Time Polymerase Chain Reaction , Herpes Simplex/diagnosis
17.
Acta odontol. latinoam ; 30(3): 109-112, 2017. ilus
Article in English | LILACS | ID: biblio-904928

ABSTRACT

In this work, we established an in vivo murine model of herpes simplex virus type 1 (HSV1) infection involving inoculation by scarification of the oral mucosain order to study its dissemination towards the trigeminal ganglion (TG). Both viral DNA and infectious virions were detected on the third day postinfection (p.i.). Viral proteins revealed by immunohistochemistry were mainly found at seven days p.i., when the latencyassociated transcript (LAT) was also detected. This model simulated the dissemination process of HSV1, which could be used to study herpes pathogenesis starting in the oral mucosa (AU)


Con el propósito de estudiar la dispersión de del Herpes Simplex Virus tipo 1 (HSV1) desde la mucosa oral hasta los ganglios trigeminales, en el presente trabajo se estableció un modelo de infección en ratones, haciendo inoculación por escarificación en la mucosa oral. Tanto ADN viral como viriones infecciosos se detectaron en los ganglios trigeminales al dia 3 postinfección (p.i.). Las proteínas virales se detectaron principalmente al día 7 p.i. cuando los transcritos asociados a latencia también fueron encontrados. El modelo de infección simula adecuadamente el proceso de dispersión del HSV1 y puede ser usado para el estudio de la patogénesis por herpes después de la infección primaria en la mucosa oral (AU)


Subject(s)
Animals , Mice , Herpesvirus 1, Human , Mouth Mucosa , Trigeminal Ganglion , Colombia , DNA, Viral , Virus Latency
18.
Rev. ecuat. pediatr ; 17(2): 30-31, 12-2016.
Article in Spanish | LILACS | ID: biblio-996581

ABSTRACT

Las enfermedades intrauterinas representan una carga importante de morbilidad para el recién nacido como es la infección por Herpes simple, al adquirir este tipo de infección puede traer diversas afecciones del sistema nervioso central. El diagnóstico diferencial de las calcificaciones en el cerebro no siempre es fácil, existen múltiples causas que pueden originar calcificaciones intracraneales, dentro de ellas causas fisiológicas congénitas, infecciosas, metabólica, vasculares y por neoplasias. Con alta mortalidad y morbilidad, la incidencia varia de 1/3.000 a 1/20.000 nacimientos. Se requiere de una sospecha clínica precoz para establecer un tratamiento adecuado, presentamos un caso de una recién nacida de 21 días de vida que presenta un cuadro febril cuyo diagnóstico fue confirmado por la técnica de reacción de la polimerasa en cadena, la misma certifico la presencia del virus del herpes simple tipo 1 en liquido céfalo raquídeo, además evidencia de calcificaciones intracraneales en tomografía axial computarizada.


Intrauterina diseases represent an important burden of morbidity for the newborn as is the infection with Herpes simplex, when aquiring this type of infeccion can bring diverse affections of the central nervous system.The differential diagnosis of calcifications in the brain is not always easy, there are multiple causes that can causse intracranial calcifications,within them congenital,infectious,metabolic,vascular and neoplastic,vascular and neoplastic causes,with high mortality and morbidity, the incidence varies from 1/3000 to 1 /20.000 briths,Early clinical suspicion is required to establish an adequate treatment,we present a case of a newborn of 21 days of life that presents a febrile condition whose diagnosis was confirmed by the polymerase chain reaction technique,it certifies the presence of herpes simplex virus type 1 in cerebrospinal fluid,in addition to evidence of intracranial calcifications in computed tomography.


Subject(s)
Humans , Female , Infant, Newborn , Acyclovir , Tomography, X-Ray Computed , Herpesvirus 1, Human , Nervous System Malformations , Congenital Abnormalities , Cerebrospinal Fluid , Herpes Simplex
19.
J. oral res. (Impresa) ; 5(4): 168-174, June 2016. tab
Article in English | LILACS | ID: biblio-982703

ABSTRACT

Abstract: objective: to detect the presence of infection by EBV (Epstein-Barr Virus), CMV (Cytomegalovirus) and HSV-1 (Herpes Simplex Virus type 1) in subgingival samples from HIV- positive patients under HAART (High Activity Antiretroviral Therapy), HIV- positive patients without HAART, HIV-negative patients with chronic periodontitis and healthy controls. Methodology: Crevicular fluid samples of 11 HIV+ patients on therapy were evaluated, 6 without antiretroviral therapy, 7 HIV- negative subjects with chronic periodontitis and 7 periodontally-healthy controls. PI (Plaque index), GI (Gingival Index), PD (probing depth) and CAL (Clinical Attachment Loss) were registered at six sites per each tooth in all teeth and subgingival plaque samples of a tooth were collected per quadrant. Nested PCR was used to detect EBV and endpoint PCR to detect infection by CMV and HSV-1. Results: Clinical parameters showed statistically significant differences between HIV-positive patients and subjects with chronic periodontitis compared with the control group (p<0.05). DNA of EBV was detected mainly in HIV-positive patients under HAART, 91 percent (10/11). DNA of CMV was detected mainly in patients without HAART, 67 percent (4/6), while HSV-1 was observed in 27 percent (3/11) of patients under HAART. In the control group no virus was detected. Coinfection was observed in 50 percent of HIV patients without HAART, 36 percent of HIV patients with HAART and 14 percent of HIV-negative with chronic periodontitis. Conclusion: Viral infection was prevalent in HIV patients under HAART and EBV was the primary viral infection detected in HIV-positive patients with chronic periodontitis.


Resumen: detectar la presencia de infección por VEB (Virus Epstein-Barr), CMV (Citomegalovirus) y VHS-1 (Virus Herpes simple tipo 1) en muestras subgingivales de pacientes VIH-positivos bajo HAART (Terapia Anti Retroviral de Alta Actividad), VIH-positivos sin HAART, pacientes VIH-negativos con periodontitis crónica y controles sanos. Metodología: Se evaluaron muestras de fluido crevicular de 11 pacientes VIH+ bajo terapia, 6 sin terapia antiretroviral, 7 sujetos VIH–negativo con periodontitis crónica y 7 controles periodontalmente sanos. Se registró el IP (Índice de placa), IG (Índice Gingival), PS (Profundidad del Sondaje) y NIC (Nivel de Inserción Clínica) en seis sitios por diente en todos los dientes y se recolectaron muestras de placa subgingival de un diente por cuadrante. Se empleó PCR anidada para detectar VEB y PCR punto final para identificar la infección con CMV y VHS-1. Resultados: Los parámetros clínicos mostraron diferencias estadísticamente significativas entre pacientes VIH-positivos y sujetos con periodontitis crónica comparados con el grupo control (p<0.05). El ADN de EBV fue detectado principalmente en pacientes VIH-positivos bajo HAART con 91 por ciento (10/11). El ADN de CMV se detectó principalmente en pacientes sin HAART, 67 por ciento (4/6), mientras que VHS-1 se observó en 27 por ciento (3/11) de los pacientes bajo HAART. En el grupo control no se detectó ningún virus. La coinfección fue observada en 50 por ciento de los pacientes VIH sin HAART, 36 por ciento de los VIH con HAART y 14 por ciento de los VIH negativos con periodontitis crónica. Conclusión: La infección viral fue predominante en los pacientes VIH bajo HAART y VEB fue la principal infección viral detectada en los pacientes VIH positivos y con periodontitis crónica.


Subject(s)
Humans , Chronic Periodontitis/virology , Cytomegalovirus/isolation & purification , Gingiva/virology , Herpesvirus 1, Human/isolation & purification , /isolation & purification
20.
Mem. Inst. Oswaldo Cruz ; 111(2): 106-113, Feb. 2016. tab, graf
Article in English | LILACS | ID: lil-772612

ABSTRACT

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.


Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Cervix Uteri/microbiology , Chlamydia trachomatis/isolation & purification , DNA, Viral/isolation & purification , Papillomaviridae/isolation & purification , Squamous Intraepithelial Lesions of the Cervix/microbiology , Uterine Cervical Neoplasms/prevention & control , Carcinogenesis , Coinfection , Cross-Sectional Studies , Cytopathogenic Effect, Viral , Cervix Uteri/pathology , Chlamydia Infections/complications , Chlamydia Infections/epidemiology , Early Detection of Cancer/methods , Epithelium/virology , Genotype , Genotyping Techniques , Herpesvirus 1, Human/isolation & purification , /isolation & purification , Molecular Typing , Mycoplasma genitalium/isolation & purification , Neisseria gonorrhoeae/isolation & purification , Papillomaviridae/classification , Papillomavirus Infections/complications , Papillomavirus Infections/epidemiology , Risk Factors , Treponema pallidum/isolation & purification , Trichomonas vaginalis/isolation & purification , Uterine Cervical Neoplasms/microbiology
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