scholarly journals Development of In vitro Tetraploid Plants of Hevea brasiliensis

2019 ◽  
pp. 1-12
Author(s):  
U. K. Divya ◽  
S. Sushama Kumari
Keyword(s):  
Hevea Brasiliensis ◽  
Chromosome Doubling ◽  
Crop Improvement ◽  
Embryo Germination ◽  
Regeneration Frequency ◽  
Immature Inflorescence ◽  
In Vitro Techniques ◽  
Flow Cytometric ◽  
Study Development

Increase in global consumption of natural rubber necessitates crop improvement of Hevea aimed at increased productivity. As conventional breeding of Hevea is very elaborate and time consuming. Hence in the present study development of tetraploids through chromosome doubling of diploid callus obtained from cultured immature inflorescence of Hevea using colchicines were attempted. Chromosome doubling of the diploid callus occurred when treated with 1.25 µM colchicine for 3 days. In higher concentrations as well as at longer exposure periods, the callus texture and viability were affected. 48 % embryo induction and a maturation frequency of 45 % were obtained. Embryo germination and plant regeneration with a germination frequency (30 %) and a regeneration frequency (20 %) were obtained. Cytological and flow cytometric analyses confirmed the tetraploid nature of the colchicines treated callus. In vitro tetraploid plant developed through these in vitro techniques can be further used in Hevea brasiliensis breeding.

scholarly journals Cryopreservation of Arecanut (Areca catechu L.) Embryogenic Calli by V Cryo-plate Method

2021 ◽  
pp. 76-82
Author(s):  
Kilingar Subrahmanya Muralikrishna ◽  
Kalathil Kundanchery Sajini ◽  
Pulikuthi Kavya ◽  
Krishna Prakash ◽  
Abdulla Abdulla Sabana ◽  
...  
Keyword(s):  
Crop Improvement ◽  
Basal Medium ◽  
Start Codon ◽  
Embryogenic Calli ◽  
Immature Inflorescence ◽  
Plate Technique ◽  
Plantlet Production ◽  
Scot Marker ◽  
Palm Species

Aims: Arecanut, a perennial palm species of Arecaceae family, has huge commercial value, and is grown mainly for its masticatory nuts. The ever-increasing demand for uniform quality plantlets from growers necessitates putting in place In vitro mass multiplication and other crop improvement programmes. The present study was carried out to standardize the procedure for cryopreservation of embryogenic calli of arecanut, derived from immature inflorescence cultures, by vitrification based cryo-plate technique. Study Design: Completely randomized design (CRD) with three replications. Place and Duration of Study: ICAR-Central Plantation Crops Research Institute, Kerala, India during 2019. Methodology: The embryogenic calli were precultured in Eeuwen's Y3 basal medium supplemented with sucrose (0.2, 0.3 and 0.4 M) for three days. Explants were affixed on cryo-plates and later dehydrated using plant vitrification solution 3 (PVS3) for 30 min. Cryoplates were inserted in cryovials and cryopreserved. Explants with no cryostorage served as control. Explants were rewarmed quickly in a water bath (40ºC) for 2 min and treated with unloading solution and cultured on recovery medium. Results: The results showed 8-10 % recovery of embryogenic calli that resulted in normal plantlet production. The clonal fidelity studies, using Start Codon Targeted (SCoT) marker, showed no variation of cryopreserved calli in comparison to the original calli. Conclusion: This preliminary study demonstrated the successful use of vitrification (V) cryo-plate technique in cryopreservation of embryogenic calli of arecanut. With better recovery percentage, the optimal concentration of sucrose in the preculture medium was found to be 0.3 M. Desiccation in PVS3 solution for 30 min had no adverse effect.

scholarly journals Development of Embryo Rescue and Shoot Regeneration Techniques in Ilex

1995 ◽  
Vol 13 (4) ◽  
pp. 164-168
Author(s):  
Pamela R. Mattis ◽  
Harry Jan Swartz ◽  
Gene Eisenbeiss
Keyword(s):  
Gibberellic Acid ◽  
Shoot Regeneration ◽  
Embryo Rescue ◽  
Colchicine Treatment ◽  
Benzyl Adenine ◽  
Embryo Germination ◽  
Indole Butyric Acid ◽  
In Vitro Techniques ◽  
Germination And Growth

Abstract Studies were conducted on embryo rescue and shoot regeneration from juvenile leaves to develop a system to improve Ilex using in vitro techniques. Unlike previous reports for Ilex, embryo germination and growth in the eight species tested was not consistently affected by light. Gibberellic acid had little effect on excised embryo germination in August through October; however, embryo germination was completely inhibited by 3 μM GA in November and December. Shoot regeneration was obtained from juvenile leaves of Ilex myrtifolia and I. opaca, the two species tested. Thidiazuron, at concentrations from 5 to 50 μM, consistently resulted in the largest percentage shoot regeneration, when compared to benzyl adenine or a lower rate of thidiazuron. Indole butyric acid or pretreatment of source shoots with cytokinin or auxin did not increase regeneration percentages. Colchicine treatment of source shoots had an effect on regeneration; 100 μM was slightly stimulatory while 5 μM was significantly inhibitory. This protocol has resulted in regeneration rates that appear suitable for production of polyploid or transformed plants.

Morphometrical estimation of fetal heart growth at different gestational ages by In vivo and In vitro techniques

2011 ◽  
Vol 3 (5) ◽  
pp. 491-494
Author(s):  
Dr. Haritha Kumari Nimmagadda ◽  
◽  
Pooja Pant Pooja Pant ◽  
Rajeev Mukhia ◽  
Dr. Aruna Mukherjee
Keyword(s):  
Fetal Heart ◽  
In Vitro Techniques

Role of 4-O Galloylchlorogenic Acid in Lung Cancer- An Insilico Approach

2017 ◽  
Vol 9 (5) ◽  
Author(s):  
Jaynthy C. ◽  
N. Premjanu ◽  
Abhinav Srivastava
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
In Silico ◽  
Computational Study ◽  
Regulation Mechanism ◽  
Down Regulation ◽  
Fruit Extract ◽  
In Vitro Techniques ◽  
Discovery Studio

Cancer is a major disease with millions of patients diagnosed each year with high mortality around the world. Various studies are still going on to study the further mechanisms and pathways of the cancer cell proliferation. Fucosylation is one of the most important oligosaccharide modifications involved in cancer and inflammation. In cancer development increased core fucosylation by FUT8 play an important role in cell proliferation. Down regulation of FUT8 expression may help cure lung cancer. Therefore the computational study based on the down regulation mechanism of FUT8 was mechanised. Sapota fruit extract, containing 4-Ogalloylchlorogenic acid was used as the inhibitor against FUT-8 as target and docking was performed using in-silico tool, Accelrys Discovery Studio. There were several conformations of the docked result, and conformation 1 showed 80% dock score between the ligand and the target. Further the amino acids of the inhibitor involved in docking were studied using another tool, Ligplot. Thus, in-silico analysis based on drug designing parameters shows that the fruit extract can be studied further using in-vitro techniques to know its pharmacokinetics.

In Vitro Screening of Azalea for Resistance to Azalea Lace Bug

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 506b-506
Author(s):  
Carol D. Robacker ◽  
S.K. Braman
Keyword(s):  
Leaf Damage ◽  
In Vitro Screening ◽  
Screening Assay ◽  
Delaware Valley ◽  
In Vitro Techniques ◽  
Culture Tube ◽  
Lace Bug ◽  
Lace Bugs ◽  
Laboratory Bioassays

Azalea lace bug (Stephanitis pyrioides) is the most serious pest on azalea. Results of laboratory bioassays and field evaluations of 17 deciduous azalea taxa have identified three resistant taxa: R. canescens, R. periclymenoides, and R. prunifolium. Highly susceptible taxa are `Buttercup', `My Mary', R. oblongifolium, and the evergreen cultivar `Delaware Valley White'. To determine whether in vitro techniques would have potential value in screening or selecting for resistance, or for the identification of morphological or chemical factors related to resistance, an in-vitro screening assay was developed. In-vitro shoot proliferation was obtained using the medium and procedures of Economou and Read (1984). Shoots used in the bioassays were grown in culture tubes. Two assays were developed: one for nymphs and one for adult lace bugs. To assay for resistance to nymphs, `Delaware Valley White' leaves containing lace bug eggs were disinfested with 70% alcohol and 20% commercial bleach, and incubated in sterile petri plates with moistened filter paper until the nymphs hatched. Five nymphs were placed in each culture tube, and cultures were incubated for about 2 weeks, or until adults were observed. To assay for resistance to adults, five female lace bugs were placed in each culture tube and allowed to feed for 5 days. Data collected on survival and leaf damage was generally supportive of laboratory bioassays and field results. Adult lace bugs had a low rate of survival on resistant taxa. Survival of nymphs was somewhat reduced on resistant taxa.

Proteomics-Based Characterization of the Effects of MMP14 on the Protein Content of Exosomes from Corneal Fibroblasts

2020 ◽  
Vol 27 (10) ◽  
pp. 979-988
Author(s):  
Kyu-Yeon Han ◽  
Jin-Hong Chang ◽  
Dimitri T. Azar
Keyword(s):  
Protein Content ◽  
Catalytic Domain ◽  
Protein Composition ◽  
Proteomics Analysis ◽  
Knock Out ◽  
Corneal Fibroblast ◽  
Flow Cytometric ◽  
Corneal Fibroblasts ◽  
Liquid Chromatography Tandem Mass

Background: Exosomes secreted by corneal fibroblasts contain matrix metalloproteinase (MMP) 14, which is known to influence pro-MMP2 accumulation on exosomes. Accordingly, we hypothesized that the enzymatic activity of MMP14 may alter the protein content of corneal fibroblast- secreted exosomes. Objective: The aim of this study was to investigate the effects of MMP14 on the composition and biological activity of corneal fibroblast-derived exosomes. Methods: Knock out of the catalytic domain (ΔExon4) of MMP14 in corneal fibroblasts was used to determine the effect of MMP14 expression on the characteristics of fibroblast-secreted exosomes. The amount of secreted proteins and their size distribution were measured using Nano Tracking Analysis. Proteins within exosomes from wild-type (WT) and ΔExon4-deficient fibroblasts were identified by liquid chromatography-tandem mass spectrometry (MS/MS) proteomics analysis. The proteolytic effects of MMP14 were evaluated in vitro via MS identification of eliminated proteins. The biological functions of MMP14-carrying exosomes were investigated via fusion to endothelial cells and flow cytometric assays. Results: Exosomes isolated from WT and ΔExon4-deficient fibroblasts exhibited similar size distributions and morphologies, although WT fibroblasts secreted a greater amount of exosomes. The protein content, however, was higher in ΔExon4-deficient fibroblast-derived exosomes than in WT fibroblast-derived exosomes. Proteomics analysis revealed that WT-derived exosomes included proteins that regulated cell migration, and ΔExon4 fibroblast-derived exosomes contained additional proteins that were cleaved by MMP14. Conclusion: Our findings suggest that MMP14 expression influences the protein composition of exosomes secreted by corneal fibroblasts, and through those biological components, MMP14 in corneal fibroblasts derived-exosomes may regulate corneal angiogenesis.

Hemostatic wound dressings: Predicting their effects by in vitro tests

2019 ◽  
Vol 33 (9) ◽  
pp. 1285-1297 ◽  
Author(s):  
Cornelia Wiegand ◽  
Martin Abel ◽  
Uta-Christina Hipler ◽  
Peter Elsner ◽  
Michael Zieger ◽  
...  
Keyword(s):  
Blood Clot ◽  
Wound Dressings ◽  
In Vitro Tests ◽  
Regenerated Cellulose ◽  
Oxidized Regenerated Cellulose ◽  
Inflammatory Reactions ◽  
In Vitro Techniques ◽  
Cotton Gauze

Background Application of controlled in vitro techniques can be used as a screening tool for the development of new hemostatic agents allowing quantitative assessment of overall hemostatic potential. Materials and methods Several tests were selected to evaluate the efficacy of cotton gauze, collagen, and oxidized regenerated cellulose for enhancing blood clotting, coagulation, and platelet activation. Results Visual inspection of dressings after blood contact proved the formation of blood clots. Scanning electron microscopy demonstrated the adsorption of blood cells and plasma proteins. Significantly enhanced blood clot formation was observed for collagen together with β-thromboglobulin increase and platelet count reduction. Oxidized regenerated cellulose demonstrated slower clotting rates not yielding any thrombin generation; yet, led to significantly increased thrombin-anti-thrombin-III complex levels compared to the other dressings. As hemostyptica ought to function without triggering any adverse events, induction of hemolysis, instigation of inflammatory reactions, and initiation of the innate complement system were also tested. Here, cotton gauze provoked high PMN elastase and elevated SC5b-9 concentrations. Conclusions A range of tests for desired and undesired effects of materials need to be combined to gain some degree of predictability of the in vivo situation. Collagen-based dressings demonstrated the highest hemostyptic properties with lowest adverse reactions whereas gauze did not induce high coagulation activation but rather activated leukocytes and complement.

scholarly journals Interleukin 8 Elicits Rapid Physiological Changes in Neutrophils That Are Altered by Inflammatory Conditions

2021 ◽  
pp. 1-17
Author(s):  
Stefan Bernhard ◽  
Stefan Hug ◽  
Alexander Elias Paul Stratmann ◽  
Maike Erber ◽  
Laura Vidoni ◽  
...  
Keyword(s):  
Systemic Inflammation ◽  
Severe Trauma ◽  
Induced Response ◽  
Neutrophil Granulocytes ◽  
Inflammatory Conditions ◽  
Flow Cytometric ◽  
Time Flow ◽  
Flow Cytometric Measurement ◽  
Detailed Kinetics

A sufficient response of neutrophil granulocytes stimulated by interleukin (IL)-8 is vital during systemic inflammation, for example, in sepsis or severe trauma. Moreover, IL-8 is clinically used as biomarker of inflammatory processes. However, the effects of IL-8 on cellular key regulators of neutrophil properties such as the intracellular pH (pH<sub>i</sub>) in dependence of ion transport proteins and during inflammation remain to be elucidated. Therefore, we investigated in detail the fundamental changes in pH<sub>i</sub>, cellular shape, and chemotactic activity elicited by IL-8. Using flow cytometric methods, we determined that the IL-8-induced cellular activity was largely dependent on specific ion channels and transporters, such as the sodium-proton exchanger 1 (NHE1) and non-NHE1-dependent sodium flux. Exposing neutrophils in vitro to a proinflammatory micromilieu with N-formyl-Met-Leu-Phe, LPS, or IL-8 resulted in a diminished response regarding the increase in cellular size and pH. The detailed kinetics of the reduced reactivity of the neutrophil granulocytes could be illustrated in a near-real-time flow cytometric measurement. Last, the LPS-mediated impairment of the IL-8-induced response in neutrophils was confirmed in a translational, animal-free human whole blood model. Overall, we provide novel mechanistic insights for the interaction of IL-8 with neutrophil granulocytes and report in detail about its alteration during systemic inflammation.

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