scholarly journals Efficient and reproducible in vitro regeneration of Solanum lycopersicum and assessment genetic uniformity using flow cytometry and SPAR methods

2017 ◽  
Vol 24 (6) ◽  
pp. 1430-1436 ◽  
Author(s):  
Abdulrahman A. Alatar ◽  
Mohammad Faisal ◽  
Eslam M. Abdel-Salam ◽  
Tomas Canto ◽  
Quaiser Saquib ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Solanum Lycopersicum ◽  
In Vitro Regeneration ◽  
Genetic Uniformity

scholarly journals In vitro regeneration and polyploidy induction in Pelargonium × hortorum L. H. Bailey

2009 ◽  
Vol 57 (5) ◽  
pp. 305-312
Author(s):  
Hana Vejsadová ◽  
Petra Kuchtová-Jadrná
Keyword(s):  
Flow Cytometry ◽  
Plant Growth ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Infiltration Method

The objective of this work was to induce in vitro shoot regeneration as influenced by plant growth re­gu­la­tors and ascertain an effective method of polyploidy induction using colchicine and oryzalin in two diploid cultivars ‘Gizela’ and ‘Black Velvet Scarlet F1’ of Pelargonium × hortorum L. H. Bailey. In both cultivars, benzyladenine (BA) significantly improved shoot regeneration compared to zeatin. The infiltration and the dip methods of explant treatment were used for polyploidy induction. Regenerants were analyzed using the flow cytometry (FCM). In ‘Gizela’ and ‘Black Velvet Scarlet F1’, 10 tetraploids on the level of 2n = 4x were found by the infiltration method. The tetraploidy was determined by the dip method in 4 shoots of ‘Gizela’ and 11 tetraploids were detected in ‘Black Velvet Scarlet F1’.

scholarly journals Patlıcanda In vitro Rejenerasyon Ortamında Farklı Orizalin ve Kolhisin Konsantrasyonlarının Tetraploid Bitki Üretimine Etkisi

2019 ◽  
Vol 7 (4) ◽  
pp. 606
Author(s):  
İlknur Çeğil ◽  
Sebahattin Çürük
Keyword(s):  
Flow Cytometry ◽  
In Vitro Regeneration ◽  
Pollen Viability ◽  
Leaf Explants ◽  
Tetraploid Plant ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Ploidy Levels ◽  
Short Shoots

The objective of this study is to obtain tetraploid plants in eggplant cultivars, Faselis (F1) and Karnaz (F1), by applying colchicine and oryzalin in in vitro regeneration medium (RM: MS with 10 µM BA and 1 µM IAA). In the study, leaf explants which have been incubated on the solidified RM for 5 or 7 days were cultured on the same medium with 2.5 or 3.75 mM colchicine for 8, 16 or 32 hours; or with 28.8 or 43.2 µM oryzalin for 12, 24 or 36 hours. Then the explants were transferred to RM without colchicine and oryzalin. Callus, buds or short shoots formed in the RM were transferred to MS medium with 0.5 μM BA to obtain long plants. The ploidy levels of regenerants were determined by flow cytometry. In Karnaz, higher tetraploid plant formation was achieved from 3.75 mM colchicine compared to 2.5 mM colchicine. The highest tetraploid plant rate was obtained by applying of 43.2 µM oryzalin for 24 hours to the explants after incubation on the RM for 7 days. Pollen viability of the tetraploid plants produced by application of colchicine and oryzalin were 76.99% and 81.19% and germination were 19.14% and 17.98%. In Faselis, tetraploid plants were obtained by applying 2.5 mM colchicine to the explants for 8 or 32 hours after incubation on RM for 7 days. However, in the oryzalin experiment, the higher tetraploid plants were produced when the explants were incubated for 5 days in the RM. Pollen viability of the tetraploid plants obtained from applications of colchicine and oryzalin were 86.41% and 95.68% and germination were 26.54% and 28.47%.

A Simple and Rapid Method to Determine GPIIb/IIIa-Receptor Inhibitor Activity in Whole Blood

1999 ◽  
Vol 19 (03) ◽  
pp. 134-138
Author(s):  
Gitta Kühnel ◽  
A. C. Matzdorff
Keyword(s):  
Flow Cytometry ◽  
Platelet Count ◽  
Blood Sample ◽  
Platelet Activation ◽  
Whole Blood ◽  
Receptor Occupancy ◽  
Aggregate Formation ◽  
Inhibitor Activity ◽  
Receptor Inhibitor

SummaryWe studied the effect of GPIIb/IIIa-inhibitors on platelet activation with flow cytometry in vitro. Citrated whole blood was incubated with increasing concentrations of three different GPIIb/IIIa-inhibitors (c7E3, DMP728, XJ757), then thrombin or ADP were added and after 1 min the sample was fixed. Samples without c7E3 but with 0.1 U/ml thrombin had a decrease in platelet count. Samples with increasing concentrations of c7E3 had a lesser or no decrease in platelet count. The two other inhibitors (DMP 725, XJ757) gave similar results. GPIIb/IIIa-inhibitors prevent aggregate formation and more single platelets remain in the blood sample. The agonist-induced decrease in platelet count correlates closely with the concentration of the GPIIb/IIIa inhibitor and receptor occupancy. This correlation may be used as a simple measure for inhibitor activity in whole blood.

In vitro regeneration of watermelon seed segments

2015 ◽  
Vol 4 (8) ◽  
Author(s):  
M. Ameri ◽  
M. Lahouti ◽  
A. Bagheri ◽  
A. Sharifi ◽  
F. Keykha Akhar
Keyword(s):  
In Vitro Regeneration ◽  
Watermelon Seed

Synthesis of a Novel Curcumin Derivative as a Potential Imaging Probe in Alzheimer’s Disease Imaging

2019 ◽  
Vol 16 (8) ◽  
pp. 723-731 ◽  
Author(s):  
Alexander Sturzu ◽  
Sumbla Sheikh ◽  
Hubert Kalbacher ◽  
Thomas Nägele ◽  
Christopher Weidenmaier ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Flow Cytometry ◽  
Transgenic Mice ◽  
Ex Vivo ◽  
Amyloid Plaques ◽  
Laser Scanning Microscopy ◽  
Β Amyloid ◽  
Curcumin Derivative

Background: Curcumin has been of interest in the field of Alzheimer’s disease. Early studies on transgenic mice showed promising results in the reduction of amyloid plaques.However, curcumin is very poorly soluble in aqueous solutions and not easily accessible to coupling as it contains only phenolic groups as potential coupling sites. For these reasons only few imaging studies using curcumin bound as an ester were performed and curcumin is mainly used as nutritional supplement. Methods: In the present study we produced an aminoethyl ether derivative of curcumin using a nucleophilic substitution reaction. This is a small modification and should not impact the properties of curcumin while introducing an easily accessible reactive amino group. This novel compound could be used to couple curcumin to other molecules using the standard methods of peptide synthesis. We studied the aminoethyl-curcumin compound and a tripeptide carrying this aminoethyl-curcumin and the fluorescent dye fluorescein (FITC-curcumin) in vitro on cell culture using confocal laser scanning microscopy and flow cytometry. Then these two substances were tested ex vivo on brain sections prepared from transgenic mice depicting Alzheimer-like β-amyloid plaques. Results: In the in vitro CLSM microscopy and flow cytometry experiments we found dot-like unspecific uptake and only slight cytotoxicity correlating with this uptake. As these measurements were optimized for the use of fluorescein as dye we found that the curcumin at 488nm fluorescence excitation was not strong enough to use it as a fluorescence marker in these applications. In the ex vivo sections CLSM experiments both the aminoethyl-curcumin and the FITC-curcumin peptide bound specifically to β- amyloid plaques. Conclusion: In conclusion we successfully produced a novel curcumin derivative which could easily be coupled to other imaging or therapeutic molecules as a sensor for amyloid plaques.

In Vitro Evaluation of Chitosan Induced Cytotoxicity against Cancerous Cell lines: MCF-7, Saos-2 and HeLa

2019 ◽  
Vol 19 ◽  
Author(s):  
Zeinab Abedian ◽  
Niloofar Jenabian ◽  
Ali Akbar Moghadamnia ◽  
Ebrahim Zabihi ◽  
Roghayeh Pourbagher ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Lines ◽  
Anticancer Agents ◽  
Fibroblast Cells ◽  
Apoptosis Induction ◽  
Cytotoxic Effects ◽  
Cancerous Cell ◽  
Significant Concentration ◽  
Mcf 7

Objective/ Background: Cancer is still the most common cause of morbidity in world and new powerful anticancer agents without severe side effects from natural sources is important. Methods: The evaluation of cytotoxicity and apoptosis induction was carried out in MCF-7,HeLa and Saos-2 as cancerous cell lines with different histological origin and human fibroblast served as control normal cell. The cells were treated with different concentrations of chitosan and the cytotoxicity was determined using MTT assay after 24, 48 and 72 h .The mode of death was evaluated by flow cytometry . Results: While both types of chitosan showed significant concentration-dependently cytotoxic effects against the three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. On the other hand, there were no significant differences between LMWC and HMWC cytotoxicity in all cell lines. The flow cytometry results showed the apoptosis pattern of death more in Saos-2 and HeLa while necrosis was more observable with MCF7. Also higher viability with both types of chitosan was seen in fibroblast as normal cells Conclusion: Chitosan shows anticancerous effect against 3 cancerous cell lines, while it is compatible with normal diploid fibroblast cells. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.

A Novel Camptothecin Derivative 3j Inhibits Nsclc Proliferation Via Induction of Cell Cycle Arrest By Topo I-Mediated DNA Damage

2019 ◽  
Vol 19 (3) ◽  
pp. 365-374 ◽  
Author(s):  
Yang Liu ◽  
Jingyin Zhang ◽  
Shuyun Feng ◽  
Tingli Zhao ◽  
Zhengzheng Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Dna Damage ◽  
Cyclin D ◽  
Dna Relaxation ◽  
Cycle Arrest ◽  
H460 Cell ◽  
H1975 Cell

Objective: The aim of this study is to investigate the inhibitory effect of camptothecin derivative 3j on Non-Small Cell Lung Cancer (NSCLCs) cells and the potential anti-tumor mechanisms. Background: Camptothecin compounds are considered as the third largest natural drugs which are widely investigated in the world and they suffered restriction because of serious toxicity, such as hemorrhagic cystitis and bone marrow suppression. Methods: Using cell proliferation assay and S180 tumor mice model, a series of 20(S)-O-substituted benzoyl 7- ethylcamptothecin compounds were screened and evaluated the antitumor activities in vitro and in vivo. Camptothecin derivative 3j was selected for further study using flow cytometry in NSCLCs cells. Cell cycle related protein cyclin A2, CDK2, cyclin D and cyclin E were detected by Western Blot. Then, computer molecular docking was used to confirm the interaction between 3j and Topo I. Also, DNA relaxation assay and alkaline comet assay were used to investigate the mechanism of 3j on DNA damage. Results: Our results demonstrated that camptothecin derivative 3j showed a greater antitumor effect in eleven 20(S)-O-substituted benzoyl 7-ethylcamptothecin compounds in vitro and in vivo. The IC50 of 3j was 1.54± 0.41 µM lower than irinotecan with an IC50 of 13.86±0.80 µM in NCI-H460 cell, which was reduced by 8 fold. In NCI-H1975 cell, the IC50 of 3j was 1.87±0.23 µM lower than irinotecan (IC50±SD, 5.35±0.38 µM), dropped by 1.8 fold. Flow cytometry analysis revealed that 3j induced significant accumulation in a dose-dependent manner. After 24h of 3j (10 µM) treatment, the percentage of NCI-H460 cell in S-phase significantly increased (to 93.54 ± 4.4%) compared with control cells (31.67 ± 3.4%). Similarly, the percentage of NCI-H1975 cell in Sphase significantly increased (to 83.99 ± 2.4%) compared with control cells (34.45 ± 3.9%) after treatment with 10µM of 3j. Moreover, increased levels of cyclin A2, CDK2, and decreased levels of cyclin D, cyclin E further confirmed that cell cycle arrest was induced by 3j. Furthermore, molecular docking studies suggested that 3j interacted with Topo I-DNA and DNA-relaxation assay simultaneously confirmed that 3j suppressed the activity of Topo I. Research on the mechanism showed that 3j exhibited anti-tumour activity via activating the DNA damage response pathway and suppressing the repair pathway in NSCLC cells. Conclusion: Novel camptothecin derivative 3j has been demonstrated as a promising antitumor agent and remains to be assessed in further studies.

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