In vitro cytotoxicity of crustacean immunostimulants for lobster (Homarus gammarus) granulocytes demonstrated using the neutral red uptake assay

2004 ◽  
Vol 17 (1) ◽  
pp. 65-73 ◽  
Author(s):  
C Hauton
Author(s):  
E Roemer ◽  
S Wittke ◽  
E TrellesSticken ◽  
JJ Piade ◽  
T Bonk ◽  
...  

AbstractThe cigarette ingredients cocoa powder, glycerol, and saccharose were investigated regarding their potential effect on the resulting mainstream smoke, i.e., smoke chemistry (Hoffmann analytes), mammalian cell cytotoxicity (Neutral Red Uptake assay), and bacterial mutagenicity (Ames assay). Each ingredient was added at three concentrations to the tobacco of a 6 mg and 10 mg ‘tar’ yield experimental American blend filter cigarette (obtained under ISO/FTC smoking regime). The lowest application concentration was equivalent to the normal approximate use level of the ingredients; the highest application level was up to 5-fold higher. The resulting data were compared with the respective control cigarettes without addition of the ingredients. The addition of cocoa powder did not lead to any consistent effects on the measured mainstream smoke analytes. Neither the in vitro cytotoxicity nor the in vitro mutagenicity was affected by cocoa addition. The addition of glycerol resulted in a decrease in the delivery of several smoke constituents (generally around 20%), e.g. aldehydes, phenolics, and N-nitrosamines. Water in the particulate phase (TPM) was distinctly increased (up to +150%). The cytotoxicity of the TPM was decreased (approx. !15%). Mutagenicity was not affected. Saccharose addition consistently increased formaldehyde delivery in smoke by up to 40% and decreased tobacco-specific N-nitrosamines by up to approximately 20%. The increase in formaldehyde is discussed in the context of the human smoker. The cytotoxicity was not affected by the addition of saccharose, while the mutagenicity of the TPM was decreased in tester strain TA98 with metabolic activation (!15%). The results are in agreement with currently available literature. Some investigations summarized in this publication are novel and have not yet been reported in the literature. Based on the total evidence, it can be concluded that the three ingredients added at their current use levels do not increase the inherent toxicity of the cigarette smoke.


2008 ◽  
Vol 3 (7) ◽  
pp. 1125-1131 ◽  
Author(s):  
Guillermo Repetto ◽  
Ana del Peso ◽  
Jorge L Zurita

Planta Medica ◽  
2018 ◽  
Vol 84 (09/10) ◽  
pp. 638-644 ◽  
Author(s):  
Sianne Schwikkard ◽  
Hannah Whitmore ◽  
Timothy Corson ◽  
Kamakshi Sishtla ◽  
Moses Langat ◽  
...  

AbstractThe Hyacinthaceae family (sensu APGII), with approximately 900 species in around 70 genera, plays a significant role in traditional medicine in Africa as well as across Europe and the Middle and Far East. The dichloromethane extract of the bulbs of Massonia pustulata (Hyacinthaceae sensu APGII) yielded two known homoisoflavonoids, (R)-5-hydroxy-3-(4-hydroxybenzyl)-7-methoxy-4-chromanone 1 and 5-hydroxy-3-(4-hydroxybenzyl)-7-methoxy-4-chromone 2 and four spirocyclic nortriterpenoids, eucosterol 3, 28-hydroxyeucosterol 4 and two previously unreported triterpenoid derivatives, (17S,23S)-17α,23-epoxy-3β,22β,29-trihydroxylanost-8-en-27,23-olide 5, and (17S, 23S)-17α,23-epoxy-28,29-dihydroxylanost-8-en-3-on-27,23-olide 6. Compounds 1, 2, 3, and 5 were assessed for cytotoxicity against CaCo-2 cells using a neutral red uptake assay. Compounds 1, 2, and 5 reduced cell viability by 70% at concentrations of 30, 100, and 100 µM, respectively. Massonia bifolia yielded three known homoisoflavonoids, (R)-(4′-hydroxy)-5-hydroxy-7-methoxy-4-chromanone 1, (R)-(4′-hydroxy)-5,7-dihydroxy-4-chromanone 7 and (R)-(3′-hydroxy-4′-methoxy)-5,7-dihydroxy-4-chromanone 9, two previously unreported homoisoflavonoids, (E)-3-benzylidene-(3′,4′-dihydroxy)-5-hydroxy-7-methoxy-4-chromanone 8 and (R)-(3′,4′-dihydroxy)-5-hydroxy-7-methoxy-4-chromanone 10, and a spirocyclic nortriterpenoid, 15-deoxoeucosterol 11. Compounds 1, 1Ac, 7, 8, 9, and 10 were screened for antiangiogenic activity against human retinal microvascular endothelial cells. Some compounds showed dose-dependent antiproliferative activity and blocked endothelial tube formation, suggestive of antiangiogenic activity.


2004 ◽  
Vol 48 (5) ◽  
pp. 1766-1772 ◽  
Author(s):  
D. L. Barnard ◽  
V. D. Hubbard ◽  
D. F. Smee ◽  
R. W. Sidwell ◽  
K. G. W. Watson ◽  
...  

ABSTRACT Picornaviruses (PV) include human rhinovirus (HRV), the primary cause of the common cold, and the enteroviruses (EV), which cause serious diseases such as poliomyelitis, meningoencephalitis, and systemic neonatal disease. Although no compounds for PV infections have been approved in the United States, pirodavir was one of the most promising capsid-binding compounds to show efficacy in human clinical trials for chemoprophylaxis of the common cold. Susceptibility to hydrolysis precluded its use as an oral agent. We have developed orally bioavailable pyridazinyl oxime ethers that are as potent as pirodavir. Compounds BTA39 and BTA188 inhibited a total of 56 HRV laboratory strains and three clinical isolates as determined by neutral red uptake assay. At concentrations of <100 nM, BTA39 inhibited 69% of the HRV serotypes and isolates evaluated, BTA188 inhibited 75%, and pirodavir inhibited 59% of the serotypes and isolates. The 50% inhibitory concentrations (IC50s) for the two compounds ranged from 0.5 nM to 6,701 nM. The compounds also inhibited EV, including coxsackie A and B viruses (IC50 = 773 to 3,608 nM) and echoviruses (IC50 = 193 to 5,155 nM). BTA39 only inhibited poliovirus strain WM-1 at 204 nM, and BTA188 only inhibited poliovirus strain Chat at 82 nM. EV 71 was inhibited by BTA39 and BTA188, with IC50s of 1 and 82 nM, respectively. Both compounds were relatively nontoxic in actively growing cells (50% cytotoxic doses, ≥4,588 nM). These data suggest that these oxime ethers warrant further investigation as potential agents for treating selected PV infections.


2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
C. M. Mattana ◽  
M. A. Cangiano ◽  
L. E. Alcaráz ◽  
A. Sosa ◽  
F. Escobar ◽  
...  

Acacia aroma, native plant from San Luis, Argentina, is commonly used as antiseptic and for healing of wounds. The present study was conducted to investigate thein vitrocytotoxicity and genotoxicity of hot aqueous extract (HAE) and ethanolic extract (EE) ofA. aroma. The cytotoxic activity was assayed by neutral red uptake assay on Vero cell. Cell treatment with a range from 100 to 5000 μg/mL of HAE and EE showed that 500 μg/mL and 100 μg/mL were the maximum noncytotoxic concentrations, respectively. The CC50was 658 μg/mL for EE and 1020 μg/mL for HAE. The genotoxicity was tested by the single-cell gel electrophoresis comet assay. The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1–20 mg/mL. The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2. At concentrations below 5 mg/mL, the results of cytotoxicity and genotoxicity of aqueous and ethanolic extracts ofAcacia aromaguarantee the safety at cell and genomic level. However further studies are needed for longer periods including animal models to confirm the findings.


1998 ◽  
Vol 26 (1) ◽  
pp. 61-68
Author(s):  
Annalaura Stammati ◽  
Franco Zampaglioni ◽  
Cristiana Zanetti

The neutral red uptake (NRU) assay was included, among others, in a validation study sponsored by the European Commission/British Home Office (EC/HO) study, for its reliability as an in vitro alternative to the Draize eye irritancy test. The test was performed in parallel by four laboratories (Istituto Superiore di Sanità [ISS], Microbiological Associates, Hatano Research Institute and Kurabo Industries) on 60 selected chemicals. The results obtained by the ISS are reported in this paper. A poor rank correlation was obtained between the in vivo endpoint and the ISS in vitro results for the full set of chemicals and for the subsets, with the exception of surfactants, by an independent statistics group. The same unsatisfactory results were obtained by the ISS group when the rank correlation was calculated for compounds divided into chemical groups. The performance of the NRU assay, as an alternative to the Draize eye irritancy test, is discussed.


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