scholarly journals Cryopreservation of 12 Vitis Species Using Apical Shoot Tips Derived from Plants Grown In Vitro

HortScience ◽  
2019 ◽  
Vol 54 (6) ◽  
pp. 976-981 ◽  
Author(s):  
Jean Carlos Bettoni ◽  
Aike Anneliese Kretzschmar ◽  
Remi Bonnart ◽  
Ashley Shepherd ◽  
Gayle M. Volk
Keyword(s):  
Cost Effective ◽  
Shoot Tips ◽  
Easy Access ◽  
Biotic And Abiotic Stresses ◽  
Breeding Programs ◽  
Vitis Species ◽  
Droplet Vitrification ◽  
Effective Manner ◽  
Ideal Method

The availability of and easy access to diverse Vitis species are prerequisites for advances in breeding programs. Plant genebanks usually maintain collections of Vitis taxa as field collections that are vulnerable to biotic and abiotic stresses. Cryopreservation has been considered an ideal method of preserving these collections as safety back-ups in a cost-effective manner. We report a droplet vitrification method used to cryopreserve 12 Vitis species (Vitis vinifera cvs. Chardonnay and ‘Riesling, V. actinifolia, V. aestivalis, V. jacquemontii, V. flexuosa, V. palmata, V. riparia, V. rupestris, V. sylvestris, V. ficifolia, V. treleasi, and V. ×novae angeliae) using shoot tips excised from plants grown in vitro. Our results demonstrated wide applicability of this technique, with regrowth levels at least 43% for 13 genotypes representing 12 Vitis species. We demonstrated that the droplet vitrification procedure can be successfully replicated by technical staff, thus suggesting that this method is ready for implementation.

scholarly journals An Artificial-Intelligence-Discovered Functional Ingredient, NRT_N0G5IJ, Derived from Pisum sativum, Decreases HbA1c in a Prediabetic Population

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1635
Author(s):  
Sweeny Chauhan ◽  
Alish Kerr ◽  
Brian Keogh ◽  
Stephanie Nolan ◽  
Rory Casey ◽  
...  
Keyword(s):  
Artificial Intelligence ◽  
Pisum Sativum ◽  
Cost Effective ◽  
Glycated Haemoglobin ◽  
Glucose Regulation ◽  
Functional Ingredient ◽  
Effective Manner ◽  
Human Skeletal Muscle Cells ◽  
Increased Risk

The prevalence of prediabetes is rapidly increasing, and this can lead to an increased risk for individuals to develop type 2 diabetes and associated diseases. Therefore, it is necessary to develop nutritional strategies to maintain healthy glucose levels and prevent glucose metabolism dysregulation in the general population. Functional ingredients offer great potential for the prevention of various health conditions, including blood glucose regulation, in a cost-effective manner. Using an artificial intelligence (AI) approach, a functional ingredient, NRT_N0G5IJ, was predicted and produced from Pisum sativum (pea) protein by hydrolysis and then validated. Treatment of human skeletal muscle cells with NRT_N0G5IJ significantly increased glucose uptake, indicating efficacy of this ingredient in vitro. When db/db diabetic mice were treated with NRT_N0G5IJ, we observed a significant reduction in glycated haemoglobin (HbA1c) levels and a concomitant benefit on fasting glucose. A pilot double-blinded, placebo controlled human trial in a population of healthy individuals with elevated HbA1c (5.6% to 6.4%) showed that HbA1c percentage was significantly reduced when NRT_N0G5IJ was supplemented in the diet over a 12-week period. Here, we provide evidence of an AI approach to discovery and demonstrate that a functional ingredient identified using this technology could be used as a supplement to maintain healthy glucose regulation.

scholarly journals In vitro Conservation and Cryopreservation of Nandina domestica, an Outdoor Ornamental Shrub

2013 ◽  
Vol 41 (2) ◽  
pp. 638 ◽  
Author(s):  
Aylin OZUDOGRU ◽  
Diogo Pedrosa Corrêa Da SILVA ◽  
Ergun KAYA ◽  
Giuliano DRADI ◽  
Renato PAIVA ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Storage Temperature ◽  
Sucrose Concentration ◽  
Aluminum Foil ◽  
Shoot Tips ◽  
In Vitro Conservation ◽  
Shoot Cultures ◽  
Storage Medium ◽  
Droplet Vitrification

The study focused on an economically-important ornamental outdoor shrub, Nandina domestica, with the aims to (i) optimize an effective in vitro conservation method, and (ii) develop a cryopreservation protocol for shoot tips by the PVS2 vitrification and droplet-vitrification techniques. For in vitro conservation of shoot cultures, the tested parameters were sucrose content in the storage medium (30, 45, 60 g/L) and storage temperature (4 °C or 8 °C). Cryopreservation was performed by applying the PVS2 vitrification solution, in 2-ml cryovials or in drops over aluminum foil strips, for 15, 30, 60 or 90 min at 0 °C, followed by the direct immersion in liquid nitrogen of shoot tips. Results show that N. domestica shoots can be conserved successfully for 6 months at both the temperatures tested, especially when 60 g/L sucrose is used in the storage medium. However, conservation at 4 °C showed to be more appropriate, as hyperhydricity was observed in post-conservation of shoots coming from storage at 8 °C. As for cryopreservation, a daily gradual increase of sucrose concentration (from 0.25 to 1.0 M) produced better protection to the samples that were stored in liquid nitrogen. Indeed, with this sucrose treatment method, a 30-min PVS2 incubation time was enough to produce, 60 days after thawing, the best recovery (47% and 50%) of shoot tips, cryopreserved with PVS2 vitrification and droplet-vitrification, respectively.

Rapid and Scalable Production of Functional Anti-Coronavirus Monoclonal Antibody CR3022 in Plants

2020 ◽  
Author(s):  
Kaewta Rattanapisit ◽  
Balamurugan Shanmugaraj ◽  
Suwimon Manopwisedjaroen ◽  
Priyo Budi Purwono ◽  
Konlavat Siriwattananon ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Developing Economies ◽  
Functional Characterization ◽  
Cost Effective ◽  
Technology Platforms ◽  
Effective Manner ◽  
Rapid Spread ◽  
Significant Public Health

Abstract Severe acute respiratory syndrome coronavirus-2 is responsible for an ongoing global outbreak of coronavirus disease (COVID-19) and represents a significant public health threat. The rapid spread of COVID-19 necessitates the development of cost-effective technology platforms for the production of diagnostic reagents/biopharmaceuticals for COVID-19. We explored the possibility of producing an anti-SARS-CoV monoclonal antibody (mAb) CR3022 and the receptor binding domain (RBD) of SARS-CoV-2 in Nicotiana benthamiana. Both RBD and the mAb were transiently expressed with the expression of 8μg/g and 130μg/g leaf fresh weight respectively. The plant-purified mAb binds to SARS-CoV-2, but fails to neutralize it in vitro. This is the first report showing the functional characterization of an anti- SARS-CoV mAb CR3022 in plants. Overall these findings showed that plants are a promising platform to produce anti-SARS-CoV mAb to use as a research reagent or a biotherapeutic in a cost-effective manner, which is especially important to developing economies during epidemics.

scholarly journals An Improved In Vitro Pyrogen Test To Detect The Presence of Endotoxin Containing Bacteria Using Limulus Amoebocyte Lysate Assay From Pharmaceutical Raw Product

1970 ◽  
Vol 1 (1) ◽  
pp. 76-79
Author(s):  
Tanvir Bashar ◽  
Khondokar Mahzebin Shurovi ◽  
Sanjida Dilshad
Keyword(s):  
Pathogenic Bacteria ◽  
Raw Materials ◽  
Cost Effective ◽  
Drug Preparation ◽  
Effective Manner ◽  
Key Word ◽  
Pseudomonas Species ◽  
Lal Assay

Most of the environmental organism possess endotoxin which presence indicates the organisms are much lethal and for the purposes bacteriological quality of pharmaceutical raw products is much important. In these current study, 2 raw materials out of 10 were found to containing pathogenic bacteria Escherichia coli and Pseudomonas species in the culture medium, that indicates the raw materials were contaminated with the deadly organism. These two raw materials checked for the presence of Endotoxin and both provided positive gel clot by Limulas amoebocyte lysate (LAL) assay. Quality maintenance and assurance is the essential need of Drug preparation in pharmaceutical sector. The result ensure that Pharmaceutical industry should need to follow GMP and HACCP to minimize the contamination for improving the biological safety of the product in a cost-effective manner. Key Word: Limulas amoebocyte lysate (LAL) assay, Gram negative bacteria, endotoxin. doi:10.3329/sjps.v1i1.1812    S. J. Pharm. Sci. 1(1&2): 76-79

scholarly journals Cryopreservation of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens) shoot tips by droplet-vitrification technique

2013 ◽  
Vol 21 (2) ◽  
pp. 79-85 ◽  
Author(s):  
Djurdjina Ružić ◽  
Tatjana Vujović ◽  
Radosav Cerović
Keyword(s):  
Liquid Medium ◽  
Room Temperature ◽  
Sucrose Concentration ◽  
Shoot Multiplication ◽  
Shoot Tips ◽  
Prunus Cerasus ◽  
Droplet Vitrification ◽  
Vitrification Solution ◽  
Gisela 5

ABSTRACT The droplet-vitrification technique was applied to in vitro shoot tips of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens). Explants were precultured in the dark at 23 °C, in liquid MS medium with a progressively increasing sucrose concentration (0.3 M for 15 h, then 0.7 M for 5 h). Loading involved a 30 min incubation of explants in a solution comprising 1.9 M glycerol and 0.5 M sucrose. Explants were dehydrated at room temperature using a solution PVS A3 [Murashige and Skoog (MS) liquid medium, 22.5% (w/v) sucrose, 37.5% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethylsulfoxide] for 30, 40 and 50 min and the PVS3 solution [MS liquid medium, 50% (w/v) sucrose, 50% (w/v) glycerol] for 60, 90 and 120 min. Explants were cooled by direct immersion in liquid nitrogen (LN) in 10 μl droplets of vitrification solution placed on aluminum foil strips. The foil strips were retrieved from LN and immersed in preheated (37 °C) unloading solution (0.8 M sucrose) for 30 s, and an equal volume of unloading solution at room temperature was added for further incubation for 30 min. Shoot tips were transferred onto the regrowth medium, cultivated in the dark for 7 days before being incubated under standard conditions. Three weeks after transferring the shoot tips onto the regrowth medium, the survival rate of control and cryopreserved explants of Gisela 5 dehydrated with PVS A3 was 100%, regardless of the treatment duration. After dehydration with solution PVS3, the survival varied between 70 and 100% for control explants and 78 and 95% for cryopreserved shoot tips. Gisela 5 shoot tips dehydrated for 40 min with PVS A3 vitrification solution demonstrated the best regrowth (38%). When using the PVS3 solution, survival of cryopreserved shoot tips was the highest (95%) after 60 min treatment followed by 40% regrowth. After three successive subcultures on shoot multiplication, medium shoots recovered viability, multiplication ability and morphology equal of that prior to cryopreservation.

A survey of current trends in near-surface electrical and electromagnetic methods

Geophysics ◽  
2006 ◽  
Vol 71 (5) ◽  
pp. G249-G260 ◽  
Author(s):  
Esben Auken ◽  
Louise Pellerin ◽  
Niels B. Christensen ◽  
Kurt Sørensen
Keyword(s):  
Precision Agriculture ◽  
Cost Effective ◽  
Geological Mapping ◽  
Easy Access ◽  
Data Sets ◽  
Near Surface ◽  
Shallow Subsurface ◽  
Effective Manner ◽  
Geotechnical Investigations ◽  
New Interpretation

Electrical and electromagnetic (E&EM) methods for near-surface investigations have undergone rapid improvements over the past few decades. Besides the traditional applications in groundwater investigations, natural-resource exploration, and geological mapping, a number of new applications have appeared. These include hazardous-waste characterization studies, precision-agriculture applications, archeological surveys, and geotechnical investigations. The inclu-sion of microprocessors in survey instruments, development of new interpretation algorithms, and easy access to powerful computers have supported innovation throughout the geophysical community and the E&EM community is no exception. Most notable are development of continuous-measurement systems that generate large, dense data sets efficiently. These have contributed significantly to the usefulness of E&EM methods by allowing measurements over wide areas without sacrificing lateral resolution. The availability of these luxuriant data sets in turn spurred development of interpretation algorithms, including: Laterally constrained 1D inversion as well as innovative 2D- and 3D-inversion methods. Taken together, these developments can be expected to improve the resolution and usefulness of E&EM methods and permit them to be applied economically. The trend is clearly toward dense surveying over larger areas, followed by highly automated, post-acquisition processing and interpretation to provide improved resolution of the shallow subsurface in a cost-effective manner.

Cryopreservation of Grapevine Shoot Tips from In Vitro Plants Using Droplet Vitrification and V Cryo-plate Techniques

2019 ◽  
pp. 469-482
Author(s):  
Jean Carlos Bettoni ◽  
Ranjith Pathirana ◽  
Remi Bonnart ◽  
Ashley Shepherd ◽  
Gayle Volk
Keyword(s):  
Shoot Tips ◽  
Droplet Vitrification ◽  
In Vitro Plants

In Vitro Toxicology and the Test Guidelines

1996 ◽  
Vol 24 (3) ◽  
pp. 435-438
Author(s):  
Kimmo Louekari
Keyword(s):  
Cost Effective ◽  
Test Methods ◽  
In Vitro Tests ◽  
In Vitro Test ◽  
In Vitro Methods ◽  
Effective Manner ◽  
Genotoxic Carcinogens ◽  
Vitro Test

Ethical, economical and scientific considerations should encourage the development of alternative and in vitro test methods. Before their adoption, in vitro methods need to be validated and scientifically justified. Demand for rigorous validation schemes for in vitro tests must be emphasised, even more than in the case of in vivo tests. The OECD has adopted in vitro guidelines for testing genotoxicity; several endpoints and mechanisms can be studied in a cost-effective manner in vitro. Similar advantages could be afforded if acute irritation and corrosion, as well as the non-genotoxic carcinogenic effects of chemicals, could be studied in vitro. Evaluation of the validation status of various methods used to study non-genotoxic carcinogens was begun by the Nordic Working Group on In Vitro Methods for Non-genotoxic Mechanisms in 1996. In some established OECD test guidelines (for example, the dermal irritation/corrosion test), there is already room for the application of in vitro methods which have not been formally validated. In January 1996, the OECD Workshop on Harmonisation of Validation and Acceptance Criteria for Alternative Toxicological Test Methods set the basis for internationally acceptable principles to be followed in the validation of in vitro test methods.

scholarly journals Effect of the Duration of Liquid Nitrogen Storage on the Regrowth of Blackberry Cryopreserved by Droplet Vitrification

2017 ◽  
Vol 66 (1-2) ◽  
pp. 44-50
Author(s):  
Tatjana Vujović ◽  
Đurđina Ružić ◽  
Radosav Cerović
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Shoot Tips ◽  
Lower Survival Rate ◽  
Droplet Vitrification ◽  
Vitrification Solution ◽  
Long Term Preservation ◽  
Direct Immersion

SummaryIn vitro shoot tips of the blackberry cultivar ‘Čačanska Bestrna’ were cryopreserved using the droplet vitrification technique. Upon loading (30 min) in a solution of 1.9 M glycerol and 0.5 M sucrose, the explants were dehydrated for 40 min on ice with the PVS A3 vitrification solution (glycerol 37.5%, dimethyl sulfoxide 15%, ethylene glycol 15% and sucrose 22.5%) and for 40 min at room temperature with the PVS3 solution (glycerol 50% and sucrose 50%). They were subsequently frozen in individual microdroplets of vitrification solution, by direct immersion in liquid nitrogen (LN), and kept therein for 2, 4, 8 and 24 h. The explant rewarming was performed in an unloading solution (0.8 M sucrose) for 30 min at room temperature. The duration of LN exposure did not exert significant effects on the survival and regrowth of explants in both types of vitrification solutions. The survival and regrowth of cryopreserved shoot tips dehydrated with PVS3 solution ranged between 90–95% and 80–90%, respectively. However, dehydration with PVS A3 resulted in a lower survival rate (80–90%) and a considerably lower regrowth rate (55–65%) of explants. Monitoring the shoots regenerated in the in vitro culture revealed their normal capacity for multiplication and rooting in comparison with the controls, which fully confirms the purpose of cryopreservation in the long-term preservation of plant material.

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