Flow cytometric analysis of the cell cycle in different coconut palm ( Cocos nucifera L.) tissues cultured in vitro

2003 ◽  
Vol 22 (1) ◽  
pp. 25-31 ◽  
Author(s):  
A. Sandoval ◽  
V. Hocher ◽  
J-L. Verdeil
Keyword(s):  
Cell Cycle ◽  
Cocos Nucifera ◽  
Flow Cytometric Analysis ◽  
Coconut Palm ◽  
Flow Cytometric ◽  
Cocos Nucifera L

New thiazacridine agents: Synthesis, physical and chemical characterization, and in vitro anticancer evaluation

2016 ◽  
Vol 36 (10) ◽  
pp. 1059-1070 ◽  
Author(s):  
MBO Chagas ◽  
NCC Cordeiro ◽  
KMR Marques ◽  
MG Rocha Pitta ◽  
MJBM Rêgo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cytotoxic Activity ◽  
Cell Cycle Progression ◽  
Antitumor Agents ◽  
Flow Cytometric Analysis ◽  
Cell Lineage ◽  
Chemical Characterization ◽  
Flow Cytometric ◽  
Physical And Chemical

A series of new thiazacridine agents were synthesized and evaluated as antitumor agents, in terms of not only their cytotoxicity but also their selectivity. The cytotoxicity assay confirmed that all compounds showed cytotoxic activity and selectivity. The new compound, 3-acridin-9-ylmethyl-5-(5-bromo-1 H-indol-3-ylmethylene)-thiazolidine-2,4-dione (LPSF/AA29 – 7a), proved to be the most promising compound as it presents lower half-maximal inhibitory concentration (IC50) values (ranging from 0.25 to 68.03 µM) depending on cell lineage. In HepG2 cells, the lowest IC50 value was exhibited by 3-acridin-9-ylmethyl-5-(4-piperidin-1-yl-benzylidene)-thiazolidine-2,4-dione (LPSF/AA36 – 7b; 46.95 µM). None of the synthesized compounds showed cytotoxic activity against normal cells (IC50 > 100 µM). The mechanism of death induction and cell cycle effects was also evaluated. Flow cytometric analysis revealed that the compounds LPSF/AA29 – 7a and LPSF/AA36 – 7b significantly increased the percentage of apoptotic cells and induced G2/M arrest in the cell cycle progression. Therefore, these new thiazacridine derivatives constitute promising antitumor agents whose cytotoxicity and selectivity properties indicate they have potential to contribute to or serve as a basis for the development of new cancer drugs in the future.

Mesenchymal Stromal Cells Expressing Interferon α Limit the Development of Acute Myeloid Leukemia, Inducing Apoptosis In Vitro and In Vivo

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5216-5216
Author(s):  
Laura M Desbourdes ◽  
Adam J Guess ◽  
Suheyla Hasgur ◽  
Kathleen M Overholt ◽  
Minjun Yu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Acute Myeloid Leukemia ◽  
Stromal Cells ◽  
Myeloid Leukemia ◽  
Flow Cytometric Analysis ◽  
Interferon Α ◽  
Flow Cytometric ◽  
Acute Myeloid

Abstract Introduction The 5-year survival for patients with acute myeloid leukemia (AML) has stagnated for over two decades at about 60% for children, 40% for young adults, and <15% for elderly patients. While most patients achieve remission, approximately 50% will relapse which is generally attributed to the persistence of leukemic stem cells. Interferon α (IFNα) is an effective therapy for patients with AML due to multiple mechanisms of action. However, high serum levels are associated with many adverse effects. In this proof-of-concept study, we used engineered mesenchymal stem/stromal cells (MSC) to deliver high concentrations of IFNα locally to an AML chloroma, potentially diminishing the poorly tolerated systemic side-effects. Methods Bone marrow MSCs from C57BL/6 mouse were isolated and transduced with a lentiviral vector expressing murine IFNα (IFNα-MSCs) and/or GFP (GFP MSCs). After measuring IFNα secretion by ELISA and confirming activity by the induction of the MHC I expression on the transduced cells, the anti-AML activity of these transduced MSCs was assessed by co-culture with the C57BL/6 AML cell line c1498 which expresses DsRed and firefly luciferase (FFluc). Apoptotic cell frequencies and cell cycle phase distributions of leukemia cells with or without MSCs were assessed by flow cytometry. The in vivo validation has been performed by subcutaneous injection of c1498 cells (chloroma) with or without GFP MSCs or IFNα MSCs in C57BL/6 mice. Tumor growth was monitored by bioluminescence imaging every 3 or 4 days. Results Flow cytometric analysis and ELISA confirmed the secretion of bio-active of IFNα by transduced MSCs (41.5 ng/1E06 MSCs/24h). In co-cultures, the presence of IFNα MSCs at the ratio 100:1 (c1498: MSC) significantly decreased the population of c1498 cells mainly by inducing apoptosis compared to MSC-free or GFP MSC co-cultures while no effect was observed on cell cycle distribution. The pro-apoptotic effect of IFNα MSCs was then investigated in vivo by subcutaneous injection of c1498 cells with or without MSCs (ratio 10:1) in C57BL/6 mice.The presence of IFNα MSCs significantly decreased leukemic cell mass as shown by the bioluminescence of the DsRed+ FFLuc+ c1498 cells. This result was confirmed by flow cytometric analysis of the percentage of DsRed + cells in the chloroma. Conclusions This study shows that IFNα MSCs present a strong anti-leukemic effect in vitro and in vivo promoting apoptosis and thus decreasing the leukemic burden. Further experiments will focus on a potential synergetic effect with Cytarabine treatment and a preclinical study using human IFNα MSCs in a xenograft murine model. Disclosures No relevant conflicts of interest to declare.

scholarly journals Effects of Surface Sterilization Protocol and Haustorium Removal on Germination and Growth of Hybrid PB121 Coconut Palm (Cocos nucifera L.) Zygotic Embryos Cultured in vitro

2020 ◽  
pp. 57-64
Author(s):  
Arnaud Agbidinoukoun ◽  
Euloge Rimson Somakpe ◽  
Serge Sètondji Houedjissin ◽  
Florent Engelmann ◽  
Corneille Ahanhanzo
Keyword(s):  
Survival Rate ◽  
Cocos Nucifera ◽  
Coconut Palm ◽  
Zygotic Embryos ◽  
High Survival ◽  
Regeneration Rate ◽  
Surface Sterilization ◽  
Number Of Leaves ◽  
Cocos Nucifera L

Aims: This study aims to identify the best surface sterilization and evaluate the effect of haustorium suppression on in vitro germination of coconut palm (Cocos nucifera L.) zygotic embryos. Study Design: Survival rate and contamination rate of zygotic embryos after different surface sterilization treatments, regeneration rate and organogenesis through the number of leaves and the length of shoots after haustorium suppression were determined. For data processing, the Analysis of Variance was used to compare the means which were separated according to Tukey test            (P = 0.05). Place and Duration of Study: Coconut fruits (hybrid PB121) were collected 12 to 14 months after controlled pollination from CRAPP (Centre de Recherches Agricoles Plantes Pérennes), station of Sèmè-kpodji in Benin. Experiments were done in Central Laboratory of Plant Biotechnology and Plant Improvement, University of Abomey-Calavi and conducted from june to december in 2019. Methodology: For the zygotic embryos surface sterilization, four treatments combining three concentrations (3%, 6% and 15%) of commercial bleach (Javel la Croix© containing 12° active chlorine) and immersion durations (5 min, 10 min and 20 min) were tested and the survival rate were determined for each treatment after two months culture. The zygotic embryos were then divided in two sets (haustorium excised embryos set and the whole embryos set) and cultured in modified Y3 medium supplemented with 7 g L-1 agar, 2.5 g L-1 activated charcoal, 5% sucrose,  6.10-3 mM 2.4 D (2.4-dichlorophonoxyacetic acid), gibberellic acid and 0.3 mM BAP(6-benzylaminopurine). After five months culture, the regeneration rate, the number of leaves and the length of shouts were recorded. Results: The high survival rate (80%) was obtained with 6% of bleach and 20 min for the immersion duration without pre-disinfection. The suppression of haustorium have significantly increased the number of leaves (4.3 ± 0.02) and the length of shoots (16.2 ±0.7cm) compared to the whole zygotic embryos. Conclusion: This protocol can help to ensure better surface sterilization of zygotic embryos before their in vitro culture and the development of vigorous plantlets in order to improve the slow growth of plantlets, when transferred to the greenhouse or field. 

scholarly journals Two-color flow-cytometric analysis of the growth cycle of Plasmodium falciparum in vitro: identification of cell cycle compartments.

1986 ◽  
Vol 34 (12) ◽  
pp. 1651-1658 ◽  
Author(s):  
J D Hare
Keyword(s):  
Cell Cycle ◽  
Plasmodium Falciparum ◽  
Dna Synthesis ◽  
Flow Cytometric Analysis ◽  
Growth Cycle ◽  
Green Fluorescence ◽  
Red Fluorescence ◽  
Color Display ◽  
Flow Cytometric

A previous study (Hare JD, Bahler DW: J Histochem Cytochem 34:215, 1986) has shown that the flow cytometric analysis of acridine-orange-stained Plasmodium falciparum growing in vitro generates a complex two-color display, regions of which correlate with the major morphological stages. In this report, four cell cycle compartments (A-D) are defined by characteristic ratios of red and green fluorescence of cells distributed throughout the erythrocytic cycle as well as by the differential effects of several metabolic inhibitors. The primary characteristic of cells in compartment A is the significant increase in red fluorescence. Inhibition of DNA synthesis by either aphidicolin or hydroxyurea causes the accumulation of cells at the interface between compartments A and B, whereas n-butyrate prevents cells in compartment A from reaching the A-B interface. Cells in compartment A display a small increase in green fluorescence which is independent of DNA synthesis but is enhanced by n-butyrate treatment. Cells in compartment B display a continued increase in red fluorescence coupled with a significant increase in green fluorescence, reflecting the onset of DNA synthesis in compartment B. The transition to compartment C is more abrupt and is associated with a marked increase in green fluorescence and little increase in red fluorescence. Compartment D is characterized by an increase in red fluorescence and a continued rise in green fluorescence. It is postulated that these discontinuities in the two-color display reflect not only changes in the rates of RNA and DNA synthesis but also decondensation of parasite chromatin in compartment A as the organism prepares for DNA synthesis, and re-condensation in compartment D as the newly replicated chromatin prepares for segregation into merozoites. The method described promises to provide a sensitive and rapid technique to study the effects of various factors on the growth cycle of the parasite.

scholarly journals Molecular diagnosis of phytoplasma transmission from zygotic embryos to in vitro regenerated plants of coconut palm (Cocos nucifera L.)

2018 ◽  
Vol 17 (27) ◽  
pp. 862-869 ◽  
Author(s):  
Wentoin Alimata Marie Pierre DARAMCOUM ◽  
Konan Jean-Louis KONAN ◽  
Saraka Didier Martial YAO ◽  
Arocha Rosete YAIMA ◽  
Eric-Blanchard Zadjéhi KOFFI ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
Cocos Nucifera ◽  
Coconut Palm ◽  
Zygotic Embryos ◽  
Regenerated Plants ◽  
Cocos Nucifera L

scholarly journals Flow cytometric analysis of in vitro bluetongue virus infection of bovine blood mononuclear cells

1992 ◽  
Vol 73 (8) ◽  
pp. 1953-1960 ◽  
Author(s):  
S. M. Barratt-Boyes ◽  
P. V. Rossitto ◽  
J. L. Stott ◽  
N. J. MacLachlan
Keyword(s):  
Virus Infection ◽  
Bluetongue Virus ◽  
Mononuclear Cells ◽  
Flow Cytometric Analysis ◽  
Bovine Blood ◽  
Flow Cytometric ◽  
Blood Mononuclear Cells

Subsitusi Fitohormon Dengan Air Kelapa (Cocos nucifera L.) pada Medium Vacin and Went Terhadap Pertumbuhan Eksplan Anggrek Dendrobium sp Secara in Vitro

2021 ◽  
Vol 3 (2) ◽  
Author(s):  
Wulan Dari Neng Gumiwang ◽  
Tintrim Rahayu ◽  
Ari Hayati
Keyword(s):  
Root Length ◽  
Cocos Nucifera ◽  
Water Concentration ◽  
Coconut Water ◽  
Culture Bottle ◽  
Randomized Design ◽  
Number Of Leaves ◽  
Completely Randomized Design ◽  
Cocos Nucifera L

The purpose of this research is to determine the concentration of young coconut water that is appropriate for the growth of orchid plantlets (Dendrobium sp.) In vitro. This study used an experimental method, descriptive data analysis to compare several different concentrations of coconut water. The design of this study uses a completely randomized design (CRD). The treatments consist of 0% coconut water concentration (as a control), 15%, 30% and 60%. Each concentration was carried out 5 replications and each repetition consisted of 5 Dendrobium sp plantlets in each culture bottle conducted for 40 HST, for observing the root length carried out for 50 HST. The highest number of shoots and leaves were produced at the same concentration, namely 150 ml / L coconut water treatment (15% concentration) with an average of 2.8 shoots and the average number of leaves 10.8 leaves. The average number of roots and the longest root length was produced at a concentration of 600 ml / L coconut water (60% concentration) with an average of 6 roots, and the longest root length was 0.5 cm.Keywords: Young coconut water, (Cocos nucifera L.), Dendrobium sp., in vitro, growth.ABSTRAKTujuan penelitian ini ialah menentukan konsentrasi air kelapa muda yang tepat untuk pertumbuhan planlet anggrek (Dendrobium sp.) secara in vitro. Penelitian ini menggunakan metode eksperimen, analisis data secara deskriptif untuk membandingan beberapa konsentrasi air kelapa yang berbeda. Rancangan penelitian ini menggunakan Rancangan Acak Lengkap (RAL). Perlakukan terdiri dari konsentrasi air kelapa 0 % (sebagai kontrol), 15% , 30% dan 60%. Masing-masing konsentrasi dilakukan 5 kali ulangan dan setiap ulangan terdiri dari 5 planlet Dendrobium sp dalam setiap botol kultur yang dilakukan selama 40 HST, untuk pengamatan panjang akar dilakukan selama 50 HST. Jumlah tunas dan jumlah daun terbanyak dihasilkan pada konsentrasi yang sama, yaitu perlakuan air kelapa 150 ml/L (konsentrasi 15%)  dengan rata-rata jumlah tunas terbanyak 2,8 tunas dan rata-rata jumlah daun terbanyak 10,8 helai daun. Rata-rata jumlah akar terbanyak dan panjang akar terpanjang dihasilkan pada konsentrasi air kelapa 600 ml/L (Konsentrasi 60%) dengan rata-rata jumlah akar terbanyak sebanyak 6 akar, dan rata-rata panjang akar terpanjang 0,5 cm.Kata kunci : Air kelapa Muda (Cocos nucifera L.), Dendrobium sp., in vitro, pertumbuhan 

Exogenous sucrose can decrease in vitro photosynthesis but improve field survival and growth of coconut (Cocos nucifera L.) in vitro plantlets

2005 ◽  
Vol 41 (1) ◽  
pp. 69-76 ◽  
Author(s):  
Gabriela Fuentes ◽  
Carlos Talavera ◽  
Carlos Oropeza ◽  
Yves Desjardins ◽  
Jorge M. Santamaria
Keyword(s):  
Cocos Nucifera ◽  
In Vitro Plantlets ◽  
Survival And Growth ◽  
Cocos Nucifera L

scholarly journals UJI DAYA HAMBAT ASAP CAIR TEMPURUNG KELAPA (Cocos nucifera L.) TERHADAP PERTUMBUHAN Escherichia coli SECARA In Vitro

2021 ◽  
Vol 9 (2) ◽  
pp. 81
Author(s):  
Rachel Daniella Dinda Maria Lumban Tobing ◽  
Made Ria Defiani ◽  
Ni Made Susun Parwanayoni
Keyword(s):  
Escherichia Coli ◽  
Cocos Nucifera ◽  
E Coli ◽  
Cocos Nucifera L

Daging yang terkontaminasi bakteri berpotensi menimbulkan penyakit yang berbahaya apabila dikonsumsi manusia. Salah  satu  kuman  khususnya bakteri  yang  mencemari  daging  baik  yang  mentah  atau  daging dengan  proses  pematangan  yang  kurang  sempurna  adalah Escherichia coli. Oleh sebab itu, E. coli perlu diminimalisir dengan cara menghambat pertumbuhannya. Salah satu cara alami untuk menghambat pertumbuhan E. coli adalah dengan uji antibakteri menggunakan asap cair. Asap cair dapat diperoleh melalui proses pirolisis dari berbagai biomassa yang mengandung selulosa, hemiselulosa, dan lignin seperti pada tempurung kelapa (Cocos nucifera L.). Asap cair mengandung senyawa fenol dan asam yang bersifat antimikroba dan antioksidan. Penelitian ini menggunakan metode sumur difusi. Analisis data menggunakan Rancangan Acak Lengkap (RAL) dengan uji ragam (ANOVA) apabila data memiliki beda nyata pada taraf uji 5% (P?5) maka dilanjutjan uji Duncan. Asap cair tempurung kelapa mampu menghambat pertumbuhan E.coli yang ditandai dengan terbentuknya zona hambat. Konsentrasi asap cair tempurung kelapa yang efektif menghambat pertumbuhan E. coli adalah konsentrasi 50% dengan diameter zona hambat 16,66 mm dan MIC pada konsentrasi 10% dengan diameter zona hambat 9,33 mm. Hasil uji fitokimia asap cair tempurung kelapa positif mengandung senyawa fenol, flavonoid, triterpenoid, dan saponin serta kadar fenol 2,403% dan kadar saponin 5,50%.

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