scholarly journals METHODS FOR PRODUCING GRAFT CHIMERAS IN VITRO

HortScience ◽  
1991 ◽  
Vol 26 (6) ◽  
pp. 756G-757 ◽  
Author(s):  
Huey-Jen Chen ◽  
Harry Jan Swartz
Keyword(s):  
Shoot Regeneration ◽  
Shoot Organogenesis ◽  
Woody Species ◽  
In Vitro Techniques ◽  
Golden Delicious ◽  
Periclinal Chimeras ◽  
Organogenesis In Vitro ◽  
Solanaceous Plants

Several authors report the synthesis of periclinal chimeras generated from graft unions of Solanaceous plants grown in the greenhouse. As this technique requires shoot organogenesis, in vitro conditions are necessary to adapt this technique to woody species. We now report several in vitro techniques necessary to mimic the in vivo graft chimera process. These include rootstock/scion preparation, micrografting and shoot organogenesis from graft unions. Zeatin and auxins have been helpful in preparing graftable material and for increasing the percentage successful grafts. A shorter exposure to organogenic medium containing thidiazuron resulted in greater percentage shoot regeneration from graft unions. Thorny/thornless Rubus and 'Liberty'/'Golden Delicious' or 'Gala' Malus (color) markers are being used to determine the percentage of these regenerants which are chimeral.

scholarly journals The effects of β-lactam antibiotics and hygromycin B on de novo shoot organogenesis in apple cv. Golden Delicious

2018 ◽  
Vol 70 (1) ◽  
pp. 179-190 ◽  
Author(s):  
Mariana Stanisic ◽  
Slavica Ninkovic ◽  
Jelena Savic ◽  
Tatjana Cosic ◽  
Nevena Mitic
Keyword(s):  
Shoot Regeneration ◽  
De Novo ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Selection Procedure ◽  
Transformation Process ◽  
Hygromycin B ◽  
Golden Delicious

Since the genetic transformation of the apple is strongly genotype-dependent and generally inefficient, the evaluation of factors affecting shoot regeneration are crucial for the establishment of a successful transformation process. In this report, we evaluated the effects of the ?-lactam antibiotics meropenem and timentin on in vitro regeneration via de novo shoot organogenesis from leaf explants of apple cv. Golden Delicious, as well as on the growth of the Agrobacterium tumefaciens strain EHA 105, and compared them with the commonly used ?-lactam cefotaxime. Also, we report for the first time the effect of hygromycin B as a selective agent in the domesticated apple, as regards shoot regeneration and shoot multiplication efficiency. We observed that cefotaxime and timentin at concentrations higher than 100 mg L-1 were sufficient to prevent Agrobacterium growth during a two-week period, while meropenem exhibited an inhibitory effect on bacterial growth at all tested concentrations (25-150 mg L-1). Cefotaxime at a concentration of 300 mg L-1 increased the number of regenerated shoots per explant (9.39) in comparison with the control (7.67). In contrast to cefotaxime, meropenem and timentin caused a decrease in shoot regeneration efficiency, but larger and more developed shoots were obtained on meropenem (25-125 mg L-1) after the same period of cultivation. Hygromycin B at a concentration of 5 mg L-1 or higher completely inhibited shoot regeneration and induced explant tissue necrosis. Therefore, the selection procedure with a final concentration of 4 mg L-1 throughout organogenesis and 10 mg L-1 for further shoot growth and multiplication is recommended for an efficient transformation process in apple cv. Golden Delicious.

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

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.

The Use of the Chorioallantoic Membrane (CAM) of the Embryonic Chick for the Direct Assessment of Implant Toxicity

1985 ◽  
Vol 13 (4) ◽  
pp. 261-266
Author(s):  
P.P. Monro ◽  
D.P. Knight ◽  
W.S. Pringle ◽  
D.M. Fyfe ◽  
J.R. Shearer
Keyword(s):  
Chorioallantoic Membrane ◽  
In Vitro Techniques ◽  
Implant Materials ◽  
Metal Foils ◽  
Complex Interactions ◽  
Cobalt Nickel ◽  
Histological Criteria ◽  
Fluid Environment

The toxicity of implant materials requires investigation prior to clinical use. We have developed a method where materials are directly applied to the chorioallantoic membrane (CAM) of 9-day-old chick embryos and toxicity is assessed using histological criteria. We evaluated the method using metal foils. The number and organisation of fibroblasts seemed to be the most useful criteria for assessing metal toxicity. Differences were greatest after 10 days of culture on the CAM. The method is sensitive enough to enable us to discriminate between the less toxic aluminium and titanium and the highly toxic cobalt, nickel and tungsten. The proposed method has advantages over in vitro techniques which provide an abnormal fluid environment and in which the more complex interactions that are possible between implant materials and tissue in vivo cannot be modelled.

scholarly journals Adipose and Muscle Cell Co-Culture System: A Novel In Vitro Tool to Mimic the In Vivo Cellular Environment

Biology ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 6
Author(s):  
Palaniselvam Kuppusamy ◽  
Dahye Kim ◽  
Ilavenil Soundharrajan ◽  
Inho Hwang ◽  
Ki Choon Choi
Keyword(s):  
Drug Development ◽  
Culture System ◽  
Cell Types ◽  
Culture Cell ◽  
In Vitro Techniques ◽  
Culture Systems ◽  
Complex Interactions ◽  
Cellular Environment

A co-culture system allows researchers to investigate the complex interactions between two cell types under various environments, such as those that promote differentiation and growth as well as those that mimic healthy and diseased states, in vitro. In this paper, we review the most common co-culture systems for myocytes and adipocytes. The in vitro techniques mimic the in vivo environment and are used to investigate the causal relationships between different cell lines. Here, we briefly discuss mono-culture and co-culture cell systems and their applicability to the study of communication between two or more cell types, including adipocytes and myocytes. Also, we provide details about the different types of co-culture systems and their applicability to the study of metabolic disease, drug development, and the role of secretory factors in cell signaling cascades. Therefore, this review provides details about the co-culture systems used to study the complex interactions between adipose and muscle cells in various environments, such as those that promote cell differentiation and growth and those used for drug development.

scholarly journals REGULATION OF CEREAL EMBRYONIC ORGANOGENESIS IN VITRO CONDITIONS

ÈKOBIOTEH ◽  
2021 ◽  
Vol 4 (1) ◽  
pp. 11-23
Author(s):  
N.N. Kruglova ◽  
Keyword(s):  
Somatic Embryogenesis ◽  
Hormonal Regulation ◽  
Zygotic Embryogenesis ◽  
Review Of The Literature ◽  
Biological Phenomenon ◽  
Organogenesis In Vitro ◽  
Cereal Embryos ◽  
Plant Embryogenesis

The article provides the brief review of the literature and own works devoted to the peculiarities of the cereal embryonic organogenesis at the early stages of ontogenesis in the conditions of in vitro culture (the so-called somatic embryogenesis, or embryoidogenesis in vitro). Particular attention is paid to the issues of hormonal regulation of the development of somatic cereal embryos from initial cells to mature structures in vitro. A comparison of somatic embryogenesis in vitro with similar events in zygotic embryogenesis in vivo confirms the validity of the principle of universality of morphogenesis processes in vivo and in vitro (Batygina, 2014). The prospects of using somatic embryogenesis in vitro as a model for studying the most complex biological phenomenon – zygotic plant embryogenesis in vivo – are discussed.

In situ and in vitro techniques for estimating degradation parameters and digestibility of diets based on maize or sorghum

2020 ◽  
Vol 158 (1-2) ◽  
pp. 150-158 ◽  
Author(s):  
B. C. Silva ◽  
M. V. C. Pacheco ◽  
L. A. Godoi ◽  
F. A. S. Silva ◽  
D. Zanetti ◽  
...  
Keyword(s):  
Dry Matter ◽  
Latin Square ◽  
In Vitro Techniques ◽  
Sorghum Grains ◽  
Sorghum Silage ◽  
The Individual ◽  
Individual Grains

AbstractAn experiment was conducted to evaluate: (1) the effects of ensiling maize or sorghum grains after reconstitution on readily soluble fraction (a), potentially degradable fraction in the rumen (b) and rate constant for degradation of b (c) of dry matter (DM), organic matter (OM) and starch (STA); and (2) an appropriate incubation time for in situ or in vitro procedures to estimate in vivo digestibility. Four rumen-cannulated Nellore bulls (body weight = 262 ± 19.6 kg) distributed in a 4 × 4 Latin square were used. Diets were based on dry ground maize (DGM); or dry ground sorghum (DGS); or reconstituted ground maize silage; or reconstituted ground sorghum silage. In vitro and in situ incubations of the individual grains and diets were simultaneously performed with in vivo digestibility. In general, reconstituted grains and diets based on reconstituted grains presented greater (P < 0.05) fraction a and lower (P < 0.05) fraction b of DM, OM and STA compared to dry grains and diets based on dry grain. However, the magnitude of response of the reconstitution and ensiling process on DM and OM degradability parameter was greater for maize than that for sorghum. Moreover, no differences (P > 0.05) were observed between DGM- and DGS-based diets for c estimates. The results suggest that the reconstitution process promotes grains protein matrix breakdown increasing STA availability. The incubation times required for in vivo digestibility estimations of DM, OM and STA are 24 h for in situ and 36 h for in vitro procedures.

scholarly journals Mycotoxins: Biotransformation and Bioavailability Assessment Using Caco-2 Cell Monolayer

Toxins ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 628
Author(s):  
Van Nguyen Tran ◽  
Jitka Viktorová ◽  
Tomáš Ruml
Keyword(s):  
Cell Monolayer ◽  
Intestinal Barrier ◽  
Intestinal Transport ◽  
Human Colon ◽  
In Vitro Models ◽  
Biologically Active ◽  
In Vitro Techniques ◽  
In Vivo Cytotoxicity

The determination of mycotoxins content in food is not sufficient for the prediction of their potential in vivo cytotoxicity because it does not reflect their bioavailability and mutual interactions within complex matrices, which may significantly alter the toxic effects. Moreover, many mycotoxins undergo biotransformation and metabolization during the intestinal absorption process. Biotransformation is predominantly the conversion of mycotoxins meditated by cytochrome P450 and other enzymes. This should transform the toxins to nontoxic metabolites but it may possibly result in unexpectedly high toxicity. Therefore, the verification of biotransformation and bioavailability provides valuable information to correctly interpret occurrence data and biomonitoring results. Among all of the methods available, the in vitro models using monolayer formed by epithelial cells from the human colon (Caco-2 cell) have been extensively used for evaluating the permeability, bioavailability, intestinal transport, and metabolism of toxic and biologically active compounds. Here, the strengths and limitations of both in vivo and in vitro techniques used to determine bioavailability are reviewed, along with current detailed data about biotransformation of mycotoxins. Furthermore, the molecular mechanism of mycotoxin effects is also discussed regarding the disorder of intestinal barrier integrity induced by mycotoxins.

scholarly journals The role of retinal photoisomerase in the visual cycle of the honeybee.

1991 ◽  
Vol 97 (1) ◽  
pp. 143-165 ◽  
Author(s):  
W C Smith ◽  
T H Goldsmith
Keyword(s):  
Visual Pigment ◽  
Compound Eye ◽  
Rod Outer Segments ◽  
Visual Cycle ◽  
In Vitro Techniques ◽  
The Third ◽  
Isomerization Processes

The compound eye of the honeybee has previously been shown to contain a soluble retinal photoisomerase which, in vitro, is able to catalyze stereospecifically the photoconversion of all-trans retinal to 11-cis retinal. In this study we combine in vivo and in vitro techniques to demonstrate how the retinal photoisomerase is involved in the visual cycle, creating 11-cis retinal for the generation of visual pigment. Honeybees have approximately 2.5 pmol/eye of retinal associated with visual pigments, but larger amounts (4-12 pmol/eye) of both retinal and retinol bound to soluble proteins. When bees are dark adapted for 24 h or longer, greater than 80% of the endogenous retinal, mostly in the all-trans configuration, is associated with the retinal photoisomerase. On exposure to blue light the retinal is isomerized to 11-cis, which makes it available to an alcohol dehydrogenase. Most of it is then reduced to 11-cis retinol. The retinol is not esterified and remains associated with a soluble protein, serving as a reservoir of 11-cis retinoid available for renewal of visual pigment. Alternatively, 11-cis retinal can be transferred directly to opsin to regenerate rhodopsin, as shown by synthesis of rhodopsin in bleached frog rod outer segments. This retinaldehyde cycle from the honeybee is the third to be described. It appears very similar to the system in another group of arthropods, flies, and differs from the isomerization processes in vertebrates and cephalopod mollusks.

Differences between the in vitro digestibility of extrusa collected from oesophageal fistulated steers and the forage consumed

2009 ◽  
Vol 49 (7) ◽  
pp. 563 ◽  
Author(s):  
David B. Coates ◽  
Robert J. Mayer
Keyword(s):  
Dry Matter ◽  
Rumen Fluid ◽  
In Vitro Digestibility ◽  
In Vitro Techniques ◽  
Pasture Legumes ◽  
Dry Matter Digestibility ◽  
Significant Difference ◽  
C3 Grasses

In a study that included C4 tropical grasses, C3 temperate grasses and C3 pasture legumes, in vitro dry matter digestibility of extrusa, measured as in vitro dry matter loss (IVDML) during incubation, compared with that of the forage consumed, was greater for grass extrusa but not for legume extrusa. The increase in digestibility was not caused by mastication or by the freezing of extrusa samples during storage but by the action of saliva. Comparable increases in IVDML were achieved merely by mixing bovine saliva with ground forage samples. Differences were greater than could be explained by increases due to completely digestible salivary DM. There was no significant difference between animals in relation to the saliva effect on IVDML and, except for some minor differences, similar saliva effects on IVDML were measured using either the pepsin–cellulase or rumen fluid–pepsin in vitro techniques. For both C4 and C3 grasses the magnitude of the differences were inversely related to IVDML of the feed and there was little or no difference between extrusa and feed at high digestibilities (>70%) whereas differences of more than 10 percentage units were measured on low quality grass forages. The data did not suggest that the extrusa or saliva effect on digestibility was different for C3 grasses than for C4 grasses but data on C3 grasses were limited to few species and to high digestibility samples. For legume forages there was no saliva effect when the pepsin–cellulase method was used but there was a small but significant positive effect using the rumen fluid–pepsin method. It was concluded that when samples of extrusa are analysed using in vitro techniques, predicted in vivo digestibility of the feed consumed will often be overestimated, especially for low quality grass diets. The implications of overestimating in vivo digestibility and suggestions for overcoming such errors are discussed.

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