scholarly journals Use of a trypan blue assay to measure the deoxyribonucleic acid content and radioactive labeling of viable cells.

1980 ◽  
Vol 28 (7) ◽  
pp. 700-703 ◽  
Author(s):  
D C Allison ◽  
P Ridolpho
Keyword(s):  
Trypan Blue ◽  
Isotopic Labeling ◽  
Blue Color ◽  
Simple Method ◽  
Viable Cells ◽  
Cellular Incorporation ◽  
Dark Blue ◽  
Thymidine Label ◽  
Selection Of

A simple method of determining cellular viability, DNA content, and isotopic labeling is described. Cells are sedimented onto slides and fixed in a mixture of trypan blue and paraformaldehyde which stains nonviable cells a dark blue color, whereas viable cells exclude the dye. The staining pattern is stable for at least 6 months and persists after the Feulgen reaction, allowing the selection of viable cells for cytophotometric DNA determinations. We ascertained cellular incorporation of a 3H-thymidine label by photomapping cells prior to cytophotometry and relocating these cells after preparation of autoradiographs. The method gives accurate quantitative DNA and labeling values of mouse thymocytes labeled in vitro and may be of value in studies of tumors containing a large proportion of necrotic cells.

A Quick and Simple Method to Evaluate Anisogramma anomala Ascospore Viability

2013 ◽  
Vol 14 (1) ◽  
pp. 22 ◽  
Author(s):  
Stephanie Heckert ◽  
Jay W. Pscheidt ◽  
Jeff K. Stone
Keyword(s):  
Cell Walls ◽  
Trypan Blue ◽  
Culture Media ◽  
Disease Incidence ◽  
Blue Staining ◽  
Simple Method ◽  
Vital Stain ◽  
Working Solution ◽  
Anisogramma Anomala ◽  
Dark Blue

Viability of ascospores of Anisogramma anomala, cause of eastern filbert blight on European hazelnut, was assessed using the vital stain trypan blue (working solution of 0.05% in lactoglycerol). Viable ascospores only had faint blue staining around their cell walls while non-viable ascospores absorbed the stain and turned dark blue. The number of viable (non-stained) ascospores as determined by trypan blue was similar to the proportion of ascospores germinating on culture media. Viability of field collected ascospores from rainwater spore traps ranged from 41 to 68%. Disease incidence of hazelnut seedlings was more closely related to differences in ascospore abundance than to differences in ascospore viability. Accepted for publication 3 January 2013. Published 9 May 2013.

scholarly journals Antitumoral activity of homeopathic and anthroposophic Viscum album's preparations: an in vitro assay

2021 ◽  
Vol 14 (2) ◽  
pp. 53-54
Author(s):  
Joao Vitor da Costa Batista ◽  
Michelle Nonato de Oliveira Melo ◽  
Adriana Passos Oliveira ◽  
Ezequiel Paulo Viriato ◽  
Stephan Baumgartner ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Anticancer Activity ◽  
Trypan Blue ◽  
Viscum Album ◽  
Mitochondrial Activity ◽  
Dependent Manner ◽  
Cytotoxic Effects ◽  
Vitro Assay ◽  
Viable Cells

Background: Viscum album (mistletoe) has been used for a variety of purposes and is widely used as an alternative treatment for cancer and other diseases. Preparations of fermented extracts from Viscum album (VA), a plant from the Loranthaceae family, have been used, mainly in European countries, with promising results as adjuvant therapies, especially in Anthroposophic Medicine.1,2,3. In vitro studies have demonstrated that various types of VA may have cytotoxic activity in carcinoma cells, being able to activate the apoptotic cascade or leading cells to necrosis. It is possible to prepare VA extracts using different procedures and solvents. The homeopathic preparations involve the use of ethanol as solvent4, while the antroposophic preparations are obtained in aqueous solvent5. Aims: The present study aimed to verify the cytotoxic effects of homeopathic and anthroposophic Viscum album preparations in human leukaemic K562 cell line. Methodology: The cytotoxic effects of anthroposophic (ISCADOR A - IA; ISCADOR P - IP; ISCADOR M - IM; ISCADOR U - IU; ISCADOR Qu - IQu) and homeopathic Viscum album extracts (VA) were evaluated in K562 cells. For this, ten independent experiments of Trypan blue assay and five independent MTT assay were carried out.6 Additionally, cell morphology was analysed by phase inverted contrast microscopy coupled to digital camera (Optika XDS-3).7 The results were expressed as mean±SD, and statistical comparisons were performed by one-way ANOVA. Results: All samples tested were able to interfere with viability and mitochondrial activity, with important morphological damage, when compared to respective controls. Considering the ISCADOR samples all of them presented similar cytotoxic activity, with lower activity attribute to IP (around 25% of viable cells) in comparison to 90% of non-viable cells induced by other samples. The homeopathic preparation originated the best results compared to anthroposophic samples. Morphological analyses supported the MTT and trypan blue assays´ results. Conclusion: The anticancer activity tested in K562 indicate that all Viscum album samples have anticancer activity, in a dose dependent manner. Further tests need to be done in order to contribute for the understanding of the Viscum album antitumoral mechanisms, as well as about differences between homeopathic and antroposophic procedures.

scholarly journals A Simple Method for Freezing Human Platelets Using 6% Dimethylsulfoxide and Storage at -80°C

Blood ◽  
1974 ◽  
Vol 43 (1) ◽  
pp. 131-136 ◽  
Author(s):  
C. R. Valeri ◽  
H. Feingold ◽  
L. D. Marchionni
Keyword(s):  
Bleeding Time ◽  
Human Platelets ◽  
Bleeding Diathesis ◽  
Simple Method ◽  
Prolonged Bleeding Time ◽  
The Mean ◽  
And Storage ◽  
Selection Of

Abstract Human platelets were preserved by freezing them with 6% DMSO in a mechanical refrigerator maintained at -80° C. The selection of 6% DMSO was arbitrary. After postthaw washing, the 51Cr recovery in vivo was about 45%, the in vitro recovery was about 75%, and the lifespan was about 8.5 days. The mean residual DMSO of 165 mg produced no adverse side effects. The number of platelets in the circulation 2 hr after the infusion of washed freeze-preserved platelets was about half that found with fresh platelets. The prolonged bleeding time induced by aspirin administration to healthy volunteers was corrected within 24 hr of transfusion of a single unit of previously frozen washed platelets; in nine of 12 studies a 50% reduction in the bleeding time occurred within 2 hr of infusion. We do not know though whether washed freeze-preserved platelets will be as effective in correcting the bleeding diathesis in thrombocytopenic patients.

Clone selection of in vitro horseradish hairy root cultures

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
R Bertóti ◽  
Á Alberti ◽  
A Böszörményi ◽  
R Könye ◽  
T Horváth ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Clone Selection ◽  
Selection Of

The Filter Loop Technique as a Method of Measuring Platelet Aggregation in the Flowing Blood of the Rat; the Inhibitory Activity of 5-oxo-l-cyclopentene-l-heptanoic Acid (AY-16,804) on Platelet Aggregation

1973 ◽  
Vol 30 (01) ◽  
pp. 138-147 ◽  
Author(s):  
Christopher R. Muirhead
Keyword(s):  
Platelet Aggregation ◽  
Prostaglandin E1 ◽  
Suitable Model ◽  
Heptanoic Acid ◽  
Simple Method ◽  
Loop Technique ◽  
Flowing Blood ◽  
Filter Loop

SummaryThe filter loop technique which measures platelet aggregation in vivo in the flowing-blood of the rat was compared to the optical density technique of Born which is carried out in vitro with platelet rich plasma. Using these two experimental models the effect on platelet aggregation of three known inhibitors sulfinpyrazone, dipyridamole and prostaglandin E1, and a novel compound 5-oxo-l-cyclopentene-l-heptanoic acid (AY-16, 804) was determined.The effects on platelet aggregation of the known inhibitors were consistent with information in the literature. Prostaglandin E1 was the most potent inhibitor in both techniques; sulfinpyrazone inhibited aggregation in both models but was less potent than prostaglandin E1. AY-16, 804 exhibited activity in vitro and in vivo similar to that of sulfinpyrazone. Dipyridamole did not inhibit platelet aggregation in vivo and did not inhibit aggregation in vitro in concentrations at which it remained soluble.The filter loop technique is a suitable model for measuring platelet aggregation in the flowing blood of the rat. It is a relatively simple method of determining aggregation and easily adapted to other species.

Rhenium-188 labeled recombinant human plasminogen kringle5 (rhk5) and preliminary biodistribution

2011 ◽  
Vol 50 (06) ◽  
pp. 234-239 ◽  
Author(s):  
R. Guo ◽  
Y. Ma ◽  
R. Zhang ◽  
S. Liang ◽  
H. Shen ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Human Serum ◽  
Critical Role ◽  
Angiogenesis Inhibitor ◽  
Simple Method ◽  
Tumour Formation ◽  
Human Plasminogen ◽  
Specificity Of Binding ◽  
A549 Lung

Summary Aim: Angiogenesis plays a critical role in tumour formation and metastasis. Suitable radiolabeled angiogenesis inhibitor can be used for noninvasive imaging of angiogenesis and radionuclide therapy. Here we prepare rhenium-188 labeled recombinant human plasminogen kringle5 (188Re-rhk5) in a convenient manner than evaluate its properties in A549 lung adenocarcinoma. Methods: 188Rerhk5 was obtained by conjugating His group at the C end of rhk5 with fac- [188Re(H2O)3(CO)3]+. Chelating efficiency of fac-[188Re(H2O)3(CO)3]+ and radiolabeling efficiency of 188Re-rhk5 were measured by radio thin-layer chromatography (RTLC). In vitro stability of 188Re-rhk5 was determined in human serum at 37°C and analyzed by RTLC. Competition test was also performed to verify the specificity of binding. A biodistribution study was carried out in nude mice bearing A549 lung adenocarcinoma. Results: 188Rerhk5 was obtained with a radiolabel efficiency of 66.1%, the radiochemical purity (RCP) can marreach 95.2% after purification. 188Re-rhk5 showed high stability in human serum, the RCP was more than 80% even 12 h after incubation. Competition test showed a high binding specificity. Furthermore, this radio-complex was excreted mainly through kidneys and showed specific tumour uptake in mice bearing A549 tumours. Conclusion: 188Re-rhk5 was prepared by a simple method. Preliminary biodistribution results showed its potential as an agent for possible tumour imaging, therapy and encouraged further investigation.

The Effects of Shock Vancomycin Concentrations on the Formation of Heteroresistance in Staphylococcus aureus

2020 ◽  
Vol 65 (9-10) ◽  
pp. 3-7
Author(s):  
V. V. Gostev ◽  
Yu. V. Sopova ◽  
O. S. Kalinogorskaya ◽  
M. E. Velizhanina ◽  
I. V. Lazareva ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
In Vitro Selection ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
High Concentration ◽  
Short Term ◽  
High Concentrations ◽  
Selection Of ◽  
Selection Of Resistance ◽  
Test Cultures

Glycopeptides are the basis of the treatment of infections caused by MRSA (Methicillin-Resistant Staphylococcus aureus). Previously, it was demonstrated that antibiotic tolerant phenotypes are formed during selection of resistance under the influence of high concentrations of antibiotics. The present study uses a similar in vitro selection model with vancomycin. Clinical isolates of MRSA belonging to genetic lines ST8 and ST239, as well as the MSSA (ATCC29213) strain, were included in the experiment. Test isolates were incubated for five hours in a medium with a high concentration of vancomycin (50 μg/ml). Test cultures were grown on the medium without antibiotic for 18 hours after each exposure. A total of ten exposure cycles were performed. Vancomycin was characterized by bacteriostatic action; the proportion of surviving cells after exposure was 70–100%. After selection, there was a slight increase in the MIC to vancomycin (MIC 2 μg/ml), teicoplanin (MIC 1.5–3 μg/ml) and daptomycin (MIC 0.25–2 μg/ml). According to the results of PAP analysis, all strains showed an increase in the area under curve depending on the concentration of vancomycin after selection, while a heteroresistant phenotype (with PAP/AUC 0.9) was detected in three isolates. All isolates showed walK mutations (T188S, D235N, E261V, V380I, and G223D). Exposure to short-term shock concentrations of vancomycin promotes the formation of heteroresistance in both MRSA and MSSA. Formation of VISA phenotypes is possible during therapy with vancomycin.

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