An in vitro alternative to the Draize eye-irritation test: Evaluation of the crystal violet staining method

1991 ◽  
Vol 5 (2) ◽  
pp. 139-143 ◽  
Author(s):  
H. Itagaki ◽  
S. Hagino ◽  
S. Kato ◽  
T. Kobayashi ◽  
M. Umeda
Keyword(s):  
Crystal Violet ◽  
Staining Method ◽  
Test Evaluation ◽  
Crystal Violet Staining ◽  
Eye Irritation ◽  
Irritation Test

Phenotypic and molecular evaluation of biofilm formation in Klebsiella pneumoniae carbapenemase (KPC) isolates obtained from a hospital of Pelotas, RS, Brazil

2021 ◽  
Vol 70 (11) ◽  
Author(s):  
Letícia Roloff Stallbaum ◽  
Beatriz Bohns Pruski ◽  
Suelen Cavalheiro Amaral ◽  
Stella Buchhorn de Freitas ◽  
Daniela Rodriguero Wozeak ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Staining Method ◽  
Multidrug Resistant ◽  
Crystal Violet Staining ◽  
Content Type ◽  
Klebsiella Pneumoniae Carbapenemase

Introduction. A significant cause of mortality in the intensive care unit (ICU) is multidrug-resistant (MDR) Gram-negative bacteria, such as Klebsiella pneumoniae carbapenemase (KPC). Biofilm production is a key factor in KPC colonization and persistence in the host, making the treatment difficult. Gap Statement. The aim of this study was to evaluate the antibiotic resistance, molecular and phenotypic biofilm profiles of 12 KPC isolates associated with nosocomial infection in a hospital in Pelotas, Rio Grande do Sul, Brazil. Methodology. Clinical isolates were obtained from different sources, identified and characterized by antibiotic resistance and carbapenemase synthesis following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Polymerase chain reaction (PCR) was used to evaluate the presence of carbapenemase (blaKPC ) and biofilm formation-associated genes (fimA, fimH, rmpA, ecpA, mrkD and wabG). Additionally, phenotypic evaluation of in vitro biofilm formation capacity was evaluated by Congo red agar (CRA) assay and the crystal violet staining method. Results. The 12 isolates evaluated in this study presented the blaKPC gene and were positive for synthesizing carbapenemases in vitro. In the carbapenem class, 83.3 % isolates were resistant and 16.7 % intermediately resistant to imipenem and meropenem. Molecular analyses found that the fimA and wabG genes were detected in 75 % of isolates, while fimH and ecpA were detected in 42 % and mrkD were detected in 8.3 % (1). The CRA assay demonstrated that all isolates were slime producers and 91.7 % (11) of isolates were classified as strong and 8.3 % (1) as moderate biofilm producers by the crystal violet staining method. The optical density (OD540nm) for strong biofilm formers ranged from 0.80±0.05 to 2.47±0.28 and was 0.55±0.12 for moderate biofilm formers. Conclusion. Our study revealed a high level of antibiotic resistance and biofilm formation in KPC isolates obtained from a hospital in Pelotas, RS, Brazil.

The CTFA evaluation of alternatives program: An evaluation of in vitro alternatives to the draize primary eye irritation test. (Phase II) oil/water emulsions

1994 ◽  
Vol 32 (10) ◽  
pp. 943-976 ◽  
Author(s):  
S.D. Gettings ◽  
L.C. Dipasquale ◽  
D.M. Bagley ◽  
P.L. Casterton ◽  
M. Chudkowski ◽  
...  
Keyword(s):  
Phase Ii ◽  
Test Phase ◽  
Eye Irritation ◽  
Irritation Test ◽  
Oil Water

Me-too validation study for in vitro eye irritation test with 3D-reconstructed human cornea epithelium, MCTT HCETM

2019 ◽  
Vol 55 ◽  
pp. 173-184 ◽  
Author(s):  
Song-E Lim ◽  
Su Jin Ha ◽  
Won-Hee Jang ◽  
Kyung-Mi Jung ◽  
Mi-Sook Jung ◽  
...  
Keyword(s):  
Validation Study ◽  
Human Cornea ◽  
Eye Irritation ◽  
Irritation Test ◽  
Cornea Epithelium

scholarly journals Antibacterial activity and mechanism of sanguinarine against Providencia rettgeri in vitro

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9543
Author(s):  
Qian Zhang ◽  
Yansi Lyu ◽  
Jingkai Huang ◽  
Xiaodong Zhang ◽  
Na Yu ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Crystal Violet ◽  
Membrane Integrity ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Crystal Violet Staining ◽  
Providencia Rettgeri ◽  
Intracellular Atp ◽  
Atp Concentration

Background Sanguinarine (SAG), a benzophenanthridine alkaloid, occurs in Papaveraceas, Berberidaceae and Ranunculaceae families. Studies have found that SAG has antioxidant, anti-inflammatory, and antiproliferative activities in several malignancies and that it exhibits robust antibacterial activities. However, information reported on the action of SAG against Providencia rettgeri is limited in the literature. Therefore, the present study aimed to evaluate the antimicrobial and antibiofilm activities of SAG against P. rettgeri in vitro. Methods The agar dilution method was used to determine the minimum inhibitory concentration (MIC) of SAG against P. rettgeri. The intracellular ATP concentration, intracellular pH (pHin), and cell membrane integrity and potential were measured. Confocal laser scanning microscopy (CLSM), field emission scanning electron microscopy (FESEM), and crystal violet staining were used to measure the antibiofilm formation of SAG. Results The MIC of SAG against P. rettgeri was 7.8 μg/mL. SAG inhibited the growth of P. rettgeri and destroyed the integrity of P. rettgeri cell membrane, as reflected mainly through the decreases in the intracellular ATP concentration, pHin and cell membrane potential and significant changes in cellular morphology. The findings of CLSM, FESEM and crystal violet staining indicated that SAG exhibited strong inhibitory effects on the biofilm formation of P. rettgeri and led to the inactivity of biofilm-related P. rettgeri cells.

scholarly journals GREEN TEA EXTRACT IN AN EYELASH GROWTH ENHANCER GEL FORMULATION: STABILITY TEST, EYE IRRITATION TEST, AND HUMAN EYELASH GROWTH ACTIVITY

2017 ◽  
Vol 10 (6) ◽  
pp. 243
Author(s):  
Indra Syahputra Roes Lie ◽  
Joshita Djajadisastra ◽  
Fadlina Chany Saputri
Keyword(s):  
Green Tea ◽  
Chorioallantoic Membrane ◽  
Test Group ◽  
Green Tea Extract ◽  
Stability Test ◽  
Eye Irritation ◽  
Before And After ◽  
Irritation Test ◽  
Tea Extract

Objective: To formulate a green tea extract (GTE), which is often used as a hair growth product, to produce an eyelash gel with good stability, effectiveness, and safety for growing eyelashes.Methods: GTE was formulated into a gel. A stability test was performed at a high temperature (40±2°C), room temperature (25±2°C), low temperature (4±2°C), and a cycling temperature. An in vitro hen’s egg test-chorioallantoic membrane assay was performed to evaluate potential eye irritation. An eyelash growth test was conducted by length measurement using an eyelash ruler before and after 2 mo of application in human volunteers. Results: The GTE gel was stable in storage at high, room, and low temperatures and at cycling temperatures and did not cause eye irritation. Eyelashes grew significantly more in the test group than in the placebo group after 2 mo of application (p<0.05). Conclusion: GTE gel provides a new, safe, and effective option for growing natural eyelashes.

Development of the short time exposure (STE) test: An in vitro eye irritation test using SIRC cells

2008 ◽  
Vol 22 (3) ◽  
pp. 760-770 ◽  
Author(s):  
Yutaka Takahashi ◽  
Mirei Koike ◽  
Hiroshi Honda ◽  
Yuichi Ito ◽  
Hitoshi Sakaguchi ◽  
...  
Keyword(s):  
Eye Irritation ◽  
Time Exposure ◽  
Irritation Test ◽  
Short Time

Evaluation of the bovine corneal opacity-permeability assay as an in vitro alternative to the Draize eye irritation test

1993 ◽  
Vol 7 (4) ◽  
pp. 471-476 ◽  
Author(s):  
Ph. Vanparys ◽  
Gh. Deknudt ◽  
M. Sysmans ◽  
G. Teuns ◽  
W. Coussement ◽  
...  
Keyword(s):  
Corneal Opacity ◽  
Eye Irritation ◽  
Irritation Test ◽  
Permeability Assay

Toxicity of Detergent-based Commercial Products on Cells of a Mouse Line in Suspension Culture as a Possible Screen for the Draize Rabbit Eye Irritation Test

1983 ◽  
Vol 11 (1) ◽  
pp. 15-21 ◽  
Author(s):  
R.B. Kemp ◽  
R.W.J. Meredith ◽  
S. Gamble ◽  
M. Frost
Keyword(s):  
In Vitro Test ◽  
Commercial Products ◽  
Eye Irritation ◽  
Rabbit Eye ◽  
Irritation Test ◽  
Vitro Test ◽  
Mild Irritants ◽  
Cells Cultured

Summary The Draize rabbit eye irritation test has several disadvantages and inadequacies when used as an indicator for potential irritancy of detergent-based commercial products. As a possible in vitro screen, it was decided to use mouse LS cells cultured in suspension, taking 50% cell death (CD50) after exposure to the product for 4h as the endpoint. This figure for 11 formulations was compared with eye irritation data ranked using an arbitrary classification of mild, moderate and severe. All samples with a CD50 less than 0.5mg/ml were severe eye irritants, while most of those with a CD50 greater than 1.0mg/ml were mild irritants. Samples between these two cytotoxicity levels were, in general, moderately irritant to the rabbit eye. It would appear that this in vitro test is a possible screen for the irritancy of detergent-based products.

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