Aims: This study was aimed to evaluate in vitro antiviral activity of topical formulations incorporated with a styrylpyrone derivative (SPD) against Herpes Simplex Virus type 1 (HSV-1). Methodology and results: Two types of SPD-incorporated formulations (ointment and gel) were tested for their antiviral activity against HSV-1 clinical strain using plaque reduction assay on Vero cells. The antiviral activity was determined based on the percentage of plaque reduction occurred between treatment and control (non-treated infected cells). In this study, 10% SPD-gel (SPD = 0.004 mg) and 20% SPD-ointment (SPD = 0.003 mg) showed plaque reduction percentage of 87% and 79% respectively. Further evaluation on the ointment base, gel base (formulation without SPD) demonstrated less than 10% of antiviral activity while pure SPD at 0.0025 mg showed 81% of plaque reduction. These results indicated that the antiviral activity observed in both SPD-incorporated ointment and gel was mainly due to SPD regardless of formulation components. Furthermore, the antiviral activities observed in both SPD-incorporated products were also in agreement with the antiviral activity observed in pure SPD. Conclusion, significance and impact study: SPD-incorporated products retained the antiviral activity and can further be tested in animal model.
Herpesvirus 1, Human

Aims: The present study is aimed at determining the antiviral activity of Eleusine indica whole plant methanol extract. Methodology and results: Whole dried plants were extracted with methanol and the solvent was evaporated using a rotary evaporator. The crude methanol extract was previously shown to have antiviral activity towards herpes simplex virus type 1 (HSV-1) with selective index (SI = CC50 / EC50) of 12.2. The extract was further studied for the possible mode of action including pretreatment, attachment, penetration or virucidal activity. The observations suggested that E. indica crude methanol extract protects cells from HSV-1 infection, inhibits virus from docking to the surface of the cells and penetrating into the cells, as well as modifying virus through the virucidal effect. Conclusion, significance and impact of study: Methanol extract of E. indica is safe with antiviral potential as a prophylactic agent, inhibits viral attachment, penetration and virucidal effect.
Herpesvirus 1, Human

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.
Animals ; Antiviral Agents ; Arsenicals ; Chlorocebus aethiops ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Sulfides ; Vero Cells

No abstract available.
Antiviral Agents* ; Herpes Simplex* ; Herpesvirus 1, Human* ; Simplexvirus*

This experiment was performed to elucidate whether the cultured rabbit corneal cells were infected with HSV-1. The direct immunofluorescent method was used to detect herpes simplex antigen in HSV-1 infected corneal epithelial cell, keratocyte and endothelial cell in vitro with fluorescein-labeled antiserum to HSV-1. Immunofluorescent localization of HSV-1 antigen was seen in all three types of corneal cells.
Endothelial Cells ; Epithelial Cells ; Herpes Simplex ; Herpesvirus 1, Human*

PURPOSE: 5-iododeoxyuridine analogues have been exclusively developed for the potential antiviral and antitumor therapeutic agents. In this study, we synthesized carbocyclic radioiododeoxyuridineanalogue (ddIVDU) and carbocyclic intermediate as efficient carbocyclic radiopharmaceuticals. MATERIALS AND METHODS: The synthesis is LAH reduction, hetero Diels-Alder reaction as key reactions including Pd(0)-catalyzed coupling reaction together with organotin. MCA-RH7777 (MCA) and MCA-tk (HSV1-tk positive) cells were treated with various concentration of carbocyclic ddIVDU, and GCV. Cytotoxicity was measured by the MTS methods. For in vitro uptake study, MCA and MCA-tk cells were incubated with 1uCi of [(125)I]carbocyclic ddIVDU. Accumulated radioactivity was measured after various incubation times. RESULTS: The synthesis of ddIVDU and precursor for radioiodination were achieved from cyclopentadiene in good overall yield, respectively. The radioiododemetallation for radiolabeling gave more than 80% yield with > 95% radiochemical purity. GCV was more toxic than carbocyclic ddIVDU in MCA-tk cells. Accumulation of [(125)I]carbocyclic ddIVDU was higher in MCA-tk cells than MCA cells. CONCLUSION: Biological data reveal that ddIVDU is stable in vitro, less toxic than ganciclovir (GCV), and selective in HSV1-tk expressed cells. Thus, this new carbocyclic nucleoside, referred to in this paper as carbocyclic 2',3'-didehydro-2',3'-dideoxy-5- iodovinyluridine (carbocyclic ddIVDU), is a potential imaging probe for HSV1-tk.
Cycloaddition Reaction ; Ganciclovir ; Herpesvirus 1, Human ; Idoxuridine ; Radioactivity ; Radiopharmaceuticals

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with HSV-1 and HSV-2 standard stain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of HSV-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and HSV-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, B and BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum from respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 42.6%) HSV was detected singly or mixed infection and 19 of the cases were HSV-2 and 1 case was HSV-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, B and BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.
Adult ; Arthritis ; Child ; Coinfection ; Diagnosis ; Digestion ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Herpesvirus 2, Human ; Herpesvirus 3, Human* ; Humans ; Joints* ; Mental Competency ; Polymerase Chain Reaction* ; Simplexvirus*

BACKGROUND: Herpes simplex virus type 1 (HSV-1) is linked specifically to oral and lip infections while HSV-2 is involved in genital infections. We evaluated a recently reported nested-multiplex ploymerase chain reaction (NM-PCR) for the detection and typing of HSV and compared the results with indirect immunofluorescence (IF) after cell culture. METHODS: One hundred thirty three specimens were received from patients suspected of having clinical HSV infections. HSV was cultured by the shell vial method and stained with type specific monoclonal antibodies. NM-PCR was performed using crude samples. RESULTS: HSV was detected in 45 (33.8%) and 46 (34.6%) of the 133 specimens by IF and NMPCR, respectively. All of the HSV IF positive specimens were also positive by NM-PCR. Typing by the two methods concurred in all but two of the 45 specimens; the two specimens were typed as HSV-1 and HSN-2, respectivey, by IF, and both as HSV-1 and HSV-2 by NM-PCR. CONCLUSIONS: Our results suggest that NM-PCR is a rapid and sensitive method for the detection and typing of HSV.
Antibodies, Monoclonal ; Cell Culture Techniques ; Fluorescent Antibody Technique, Indirect* ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Lip ; Simplexvirus*

The development of a multiplex polymerase chain reaction (PCR) method for rapid and accurate detection and typing of herpes simplex virus type 1 (HSV-1), and type-2 (HSV-2), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) is very important for clinical diagnosis to allow the deliver of therapy as early as possible. Large scale amplifications by multiplex PCR of viral DNA can lower the cost and time for viral diagnosis. In this study, therefore sensitive quadruplex PCR was achieved by optimizing parameters such as primers, and 1.5 mM magnesium and 200 uM dNTPs concentrations. The concentrations of HSV-1, HSV-2, CMV and EBV primers were 0.5, 0.3, 0.25 and 0.25 pmoles, respectively. Optimal annealing temperature was 54 degrees C. Employing these conditions, we could detect 10 copies of reconstructed template plasmid DNA, which were cloned to vectors containing target sequences of viral DNA. PCR products of 271 bp for HSV-1, 231 bp for HSV-2, 368 bp for CMV, and 326 bp for EBV were separated on 5.0% polyacrylamide gel electrophoresis and confirmed by direct sequencing. The present study showed that the quadruplex PCR assay described herein has potential application in clinical diagnosis, when rapid, accurate detection and typing of viruses HSV-1, HSV-2, CMV or EBV are necessary.
Cytomegalovirus/classification/*isolation & purification ; Herpesvirus 1, Human/classification/*isolation & purification ; Herpesvirus 2, Human/classification/*isolation & purification ; Herpesvirus 4, Human/classification/*isolation & purification ; Human ; *Polymerase Chain Reaction/methods ; Sensitivity and Specificity ; Support, Non-U.S. Gov't

Herpes simplex virus (HSV) is a common causative agent of genital ulceration and can lead to subsequent neurological disease in some cases. Here, using a genital infection model, we tested the efficacy of vinegar-processed flos of Daphne genkwa (vp-genkwa) to modulate vaginal inflammation caused by HSV-1 infection. Our data revealed that treatment with optimal doses of vp-genkwa after, but not before, HSV-1 infection provided enhanced resistance against HSV-1 infection, as corroborated by reduced mortality and clinical signs. Consistent with these results, treatment with vp-genkwa after HSV-1 infection reduced viral replication in the vaginal tract. Furthermore, somewhat intriguingly, treatment of vp-genkwa after HSV-1 infection increased the frequency and absolute number of CD3-NK1.1+NKp46+ natural killer (NK) cells producing interferon (IFN)-gamma and granyzme B, which indicates that vp-genkwa treatment induces the activation of NK cells. Supportively, secreted IFN-gamma was detected at an increased level in vaginal lavages of mice treated with vp-genkwa after HSV-1 infection. These results indicate that enhanced resistance to HSV-1 infection by treatment with vp-genkwa is associated with NK cell activation. Therefore, our data provide a valuable insight into the use of vp-genkwa to control clinical severity in HSV infection through NK cell activation.
Animals ; Daphne* ; Herpesvirus 1, Human ; Inflammation ; Interferons ; Killer Cells, Natural* ; Mice ; Mortality ; Simplexvirus ; Therapeutic Irrigation ; Ulcer