A colorimetric assay for the measurement of the sensitivity of herpes simplex viruses to antiviral agents

McLaren, Colin; Ellis, M N; Hunter, G.A.

doi:10.1016/0166-3542(83)90001-3

Cited by 140 publications

(62 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Antiviral activities were also determined by monitoring the ability of ACV, A111OU, or both to prevent the cytopathic effects of HSV-1 and HSV-2. Cell viability was assessed by the dye-uptake method (25). In these experiments, submicromolar (noninhibitory) concentrations of A111OU significantly decreased the IC50 of ACV for both types of virus (J. Hill, personal communication).…”

Section: Methodsmentioning

confidence: 99%

2-Acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (A1110U), a potent inactivator of ribonucleotide reductases of herpes simplex and varicella-zoster viruses and a potentiator of acyclovir.

Spector

Harrington²,

Morrison³

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

(9) and to potentiate the antiherpetic activities of ACV. In addition to producing the expected decrease in the dGTP pool, A723U also caused the pool of ACVTP to increase by 10-fold (17).In the present study, the properties of 2-acetylpyridine 5-[(dimethylamino)thiocarbonyl]thiocarbonohydrazone (A111OU) were investigated. This compound was found to be considerably more potent than A723U. At submicromolar concentrations, A111OU inactivated the ribonucleotide reductases of these three herpes viruses. It also markedly potentiated the activity of ACV against the viruses in vitro. Furthermore, as reported elsewhere ( §, ¶), A111OU and ACV produced significantly synergistic therapy as topical treatment of HSV-infected animals. MATERIALS AND METHODSReagents. All reagents not described below were obtained as previously described (11).Synthesis of A111OU. A mixture of 24.2 g (0.203 mol) of 4,4-dimethylthiosemicarbazide, 26.6 g (0.220 mol) of 2-acetylpyridine, 2.5 ml of glacial acetic acid, and 100 ml of 95% (vol/vol) ethanol was refluxed for 1.5 hr and then incubated overnight at room temperature. Thick orange needles contaminated with a small amount of a light-colored solid were collected, washed with 20 ml of ethanol, and, while still damp, added to 450 ml of boiling methanol. After 10 min, the mixture was filtered. The undissolved solid was re-incubated with 50 ml of boiling methanol for 3 min and then filtered. After 15 min at room temperature, the yellow product recrystallized from the combined filtrates and was collected by filtration.

show abstract

Section: Methodsmentioning

confidence: 99%

2-Acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (A1110U), a potent inactivator of ribonucleotide reductases of herpes simplex and varicella-zoster viruses and a potentiator of acyclovir.

Spector

Harrington²,

Morrison³

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Cytotoxicity evaluation by cell viability was performed by the neutral red dye method (McLaren et al 1983) on 3T3 as described in Bazes et al (2006). Cellular suspensions (3,5× 10 5 3T3 cells mL −1 purchased from Eurobio) were incubated with various concentrations of algae extracts and biocides (10-300 μg mL −1 , 4 wells per concentration) in 96-well plates (72 h, 37°C, 5%CO 2 ) in Eagle's MEM 10% FCS.…”

Section: Toxicitymentioning

confidence: 99%

Investigation of the antifouling constituents from the brown alga Sargassum muticum (Yendo) Fensholt

et al. 2008

View full text Add to dashboard Cite

One of the most promising alternatives to toxic heavy metal-based paints is offered by the development of antifouling coatings in which the active ingredients are compounds naturally occurring in marine organisms and operating as natural antisettlement agents. Sessile marine macroalgae are remarkably free from settlement by fouling organisms. They produce a wide variety of chemically active metabolites in their surroundings, potentially as an aid to protect themselves against other settling organisms. In this study, a dichloromethane extract from the brown seaweed Sargassum muticum was tested in situ and, after 2 months of immersion, showed less fouling organisms on paints in which the extract was included, compared to paints containing only copper after 2 months of immersion. No barnacles or mussels have been observed on the test rack. Identification by NMR and GC/MS of the effective compound revealed the abundance of palmitic acid, a commonly found fatty acid. Pure palmitic acid showed antibacterial activity at 44 µg mL −1, and also inhibited the growth of the diatom Cylindrotheca closterium at low concentration (EC 50 =45.5 µg mL −1 ), and the germination of Ulva lactuca spores at 3 µg mL −1 . No cytotoxicity was highlighted, which is promising in the aim of the development of an environmentally friendly antifouling paint.

show abstract

“…Cells were examined daily under a phase-contrast microscope to determine the minimum concentration that induced alterations in cell morphology, including swelling, shrinkage, granularity and detachment. Cytotoxicity was tested using the neutral red dye method (McLaren et al 1983). After shaking, optical density (OD) was measured at 540 nm using a spectrophotometer (SpectraCount Anti-HSV-1 assays by cell viability.…”

Section: Methodsmentioning

confidence: 99%

“…After incubation, antiviral activity was evaluated by the neutral red dye method (McLaren et al 1983). Cell and virus controls were carried out simultaneously, and 3 replicates were performed using 3 different oyster homogenates.…”

Section: Methodsmentioning

confidence: 99%

In vitro research of anti-HSV-1 activity in different extracts from Pacific oysters Crassostrea gigas

Olicard¹,

Didier²,

Marty³

et al. 2005

Dis. Aquat. Org.

View full text Add to dashboard Cite

Mortalities related to the detection of Ostreid Herpesvirus 1 (OsHV-1) have been previously reported in France among larvae and spat of the Pacific oyster Crassostrea gigas. Adult oysters appear less sensitive to herpesvirus infections, although OsHV-1 has been detected in adults without signs of disease or mortality. This suggests that the virus is able to persist in its host and that adult oysters may be able to control OsHV-1 infection. Little is known about antiviral substances in invertebrates. The present work concerns the research of antiviral substances in adult oyster C. gigas, where putative antiviral activities were monitored using 3 strategies: (1) in metabolites with variable polarity, (2) in peptidic extracts and (3) in crude haemolymph. In vitro antiviral assays were based on inhibition of Herpes simplex virus type 1 (HSV-1) replication in Vero cell monolayers. All extracts presented no cytotoxicity. Antiviral activity was detected in the fresh filtered haemolymph (EC 50 :425 µg ml -1) and seasonal variation of the haemolymph antiviral activity was monitored.KEY WORDS: Oyster · Crassostrea gigas · Antiviral defence · OsHV-1 · HSV-1 · Antiviral peptides · Haemolymph Resale or republication not permitted without written consent of the publisherDis Aquat Org 67: [141][142][143][144][145][146][147] 2005 reported (Le Deuff et al. 1996). Adult oysters could be more resistant to viral infections in developing effective immunity against OsHV-1.Despite the impact that viral diseases have on shellfish cultures, little is known about how shellfish farmers can prevent and treat viral infections or about how shellfish control viral diseases. Although invertebrates do not produce specific antibodies (Shapiro 1975), they may rely on their innate defence for host protection against viruses. Reports on viruses infecting bivalves are available today, but those focusing on antiviral defence mechanisms are poorly documented. Antiviral substances have rarely been reported in bivalves and are particularly rare for Crassostrea gigas (Prescott et al. 1966, Li & Traxler 1972, Bachère et al. 1989, Lee & Maruyama 1998.The main objective of the present study was the detection of antiviral substances in adult Crassostrea gigas oysters. Antiviral and cytotoxicity activities of oyster aqueous and alcoholic extracts were evaluated in vitro against Herpes simplex virus type 1 (HSV-1) infecting Vero cells. Cytotoxicity activity assays were performed in order to demonstrate that oyster extracts were not causing Vero cell mortality. The protocol used in the present study to prepare aqueous and alcoholic oyster extracts was previously developed for extraction of marine bioactive substances (Hellio et al. 2000). Antiviral activity of oyster peptide extracts and crude haemolymph was also monitored. Peptide extraction from oyster tissues was developed on the basis of previous works on peptidic antimicrobial substances from frogs (Amiche et al. 2000, Matutte et al. 2000, lobsters (Nousiainen et al. 1998), and mussels (Mitta ...

show abstract

A colorimetric assay for the measurement of the sensitivity of herpes simplex viruses to antiviral agents

Cited by 140 publications

References 14 publications

2-Acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (A1110U), a potent inactivator of ribonucleotide reductases of herpes simplex and varicella-zoster viruses and a potentiator of acyclovir.

2-Acetylpyridine 5-[(dimethylamino)thiocarbonyl]-thiocarbonohydrazone (A1110U), a potent inactivator of ribonucleotide reductases of herpes simplex and varicella-zoster viruses and a potentiator of acyclovir.

Investigation of the antifouling constituents from the brown alga Sargassum muticum (Yendo) Fensholt

In vitro research of anti-HSV-1 activity in different extracts from Pacific oysters Crassostrea gigas

Contact Info

Product

Resources

About