scholarly journals Explants selection and conditions of the donor material cultivation for Callistephus Chinensis (L.) Ness. introduced sorts in vitro introduction with a view to its further use in greening

2019 ◽  
pp. 85-90
Author(s):  
S. Turchyna
Keyword(s):  
Vegetative Reproduction ◽  
Parameters Optimization ◽  
High Quality ◽  
Physical Conditions ◽  
Donor Material ◽  
Planting Material ◽  
Research Material ◽  
Literary Sources

The aim. The technology of cultivating high-quality planting material of Callistephus Chinensis (Callistephus Chinensis (L.) Nees) in vitro has been developed on the basis of planting material parameters optimization taking into account its biological characteristics and its elements have been improved. In particular, a set of biotechnological techniques for the production of self-adhesives, regeneration plants and their adaptation to in vivo conditions have been developed. The data of literary sources of Ukrainian and foreign origins on the reproduction of varieties (Callistephus Chinensis (L.) Nees) in vitro and in notional conditions are studied and generalized. Biotechnology methods, microclonal reproduction in particular, along with fundamental researches, have been widely used in the applied directions of experimental biology. First of all, it is about the plants gene pool preservation, creating the high-quality gardening material and accelerated vegetative reproduction. Research material and methods. 20 varieties of Chinensis Callistephus plants with different important characteristics, origin and application direction were used in our studies. Research results. Six genotypes were selected as a result of studies on the seeds germination of the most decorative varieties of Callistephus Chinensis with different economic and valuable characteristics. These were selected for in vitro introduction in order to accelerate their reproduction. Discussion. For this purpose we collected previously selected 100 seeds in laboratory conditions, at a temperature of 18–20 oC and humidity of 75–80 %. The seeds were collected in a phased manner, namely 10 pieces each and placed in a flask with distilled water for a period of up to 20–30 minutes. After a period of time, seeds with an incomplete germ bag floated up to the surface of the flask and made, respectively, 10 to 30 % for different genotypes of. Conclusions. That is, the optimal physical conditions for donor material cultivation have been selected and the mechanism for selecting the filled seed germs of the studied genotypes has been worked out. Key words: source material, Callistephus Chinensis, varieties, introduction, in vitro, economically valuable signs, genotype.

Miscanthus: genetic diversity and a method of ploidy variability identification using fluorescent cytophotometry

2017 ◽  
Vol 4 (3) ◽  
pp. 19-27
Author(s):  
N. Kovalchuk ◽  
M. Roik
Keyword(s):  
Genetic Diversity ◽  
Gene Pool ◽  
Vegetative Reproduction ◽  
Bioenergy Crops ◽  
Miscanthus Sinensis ◽  
Planting Material ◽  
Miscanthus Giganteus ◽  
Genome Ploidy

Aim. Due to the introduction of the Miscanthus species, attributed to the European gene pool, in Ukraine, it is necessary to develop methods for the determination of genome ploidy and adjust them to the foreign methods in order to ensure high purity of the planting material, to study genetic diversity, to produce new polyploid lines and select alternative Miscanthus × giganteus clones (3x). Methods. Cytological, biotechnological, fl uores- cence cytophotometry, fi eld, laboratory. Results. Domestic diploid millet (Panicum) variety of Veselopodilska Research Breeding Station and grain sorghum (Sorghum) variety Dniprovsky, whose number of chromosomes was previously investigated, served as standard genotypes for the ploidy identifi cation with Partec ploidy analyser (Germany). Using the technique, various species of miscanthus, namely Miscanthus × giganteus (3x), Miscanthus sinensis (2x), and Miscanthus saccharifl orus (2x) were selected and multiplied by clones. The heterogeneity of the Miscanthus × giganteus (3x) population of the two ecotypes was determined based on the level of genome ploidy in the vegetative reproduction of rhizomes which originated from Poland and Austria. Conclusions. Due to the complexity of cytological research, the need to involve the representatives of the Miscanthus genus in the development of bioenergy in Ukraine, and to differentiate them both in vivo and in vitro conditions to assimilate the European gene pool, a new methodology for identifi cation of plant material of different miscanthus species using the method of fl uorescence cytophotometry is presented. The ploidy of commercial foreign samples of miscanthus, introduced in the network of research and selection stations of the Institute of Bioenergy Crops and Sugar Beets of NAAS, was identifi ed.

scholarly journals Optimization of X-ray Investigations in Dentistry Using Optical Coherence Tomography

Sensors ◽  
2021 ◽  
Vol 21 (13) ◽  
pp. 4554
Author(s):  
Ralph-Alexandru Erdelyi ◽  
Virgil-Florin Duma ◽  
Cosmin Sinescu ◽  
George Mihai Dobre ◽  
Adrian Bradu ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Radiation Dose ◽  
Imaging Techniques ◽  
Image Resolution ◽  
Optical Coherence ◽  
X Rays ◽  
High Quality ◽  
X Ray

The most common imaging technique for dental diagnoses and treatment monitoring is X-ray imaging, which evolved from the first intraoral radiographs to high-quality three-dimensional (3D) Cone Beam Computed Tomography (CBCT). Other imaging techniques have shown potential, such as Optical Coherence Tomography (OCT). We have recently reported on the boundaries of these two types of techniques, regarding. the dental fields where each one is more appropriate or where they should be both used. The aim of the present study is to explore the unique capabilities of the OCT technique to optimize X-ray units imaging (i.e., in terms of image resolution, radiation dose, or contrast). Two types of commercially available and widely used X-ray units are considered. To adjust their parameters, a protocol is developed to employ OCT images of dental conditions that are documented on high (i.e., less than 10 μm) resolution OCT images (both B-scans/cross sections and 3D reconstructions) but are hardly identified on the 200 to 75 μm resolution panoramic or CBCT radiographs. The optimized calibration of the X-ray unit includes choosing appropriate values for the anode voltage and current intensity of the X-ray tube, as well as the patient’s positioning, in order to reach the highest possible X-rays resolution at a radiation dose that is safe for the patient. The optimization protocol is developed in vitro on OCT images of extracted teeth and is further applied in vivo for each type of dental investigation. Optimized radiographic results are compared with un-optimized previously performed radiographs. Also, we show that OCT can permit a rigorous comparison between two (types of) X-ray units. In conclusion, high-quality dental images are possible using low radiation doses if an optimized protocol, developed using OCT, is applied for each type of dental investigation. Also, there are situations when the X-ray technology has drawbacks for dental diagnosis or treatment assessment. In such situations, OCT proves capable to provide qualitative images.

scholarly journals Investigating the Intrafollicular Factors Affecting Oocyte Developmental Competency

2021 ◽  
Author(s):  
◽  
Zaramasina Clark
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Significant Finding ◽  
High Quality ◽  
Final Maturation

<p>The number of cycles of assisted reproductive technologies (ART) performed increased by ~9.5 % globally between 2008 and 2010. In spite of this, the success rate in terms of delivery was only ~19.0 % (Dyer et al., 2016). This discrepancy between the demand for, and success of, these technologies necessitates the development of tools to improve ART efficiency. To facilitate this, a better understanding of how the microenvironment changes within the developing follicle to culminate in a mature, developmentally-competent oocyte is required. This study employed an in vivo and in vitro ovine model to investigate the relationship between the surrounding microenvironment and oocyte maturation, and in particular, the attainment of oocyte developmental competency and high-quality embryos.  The first objective of this PhD study was to comprehensively investigate the changing microenvironment of in vivo matured, presumptive preovulatory (PPOV) follicles from wild-type (++) and high ovulation rate (OR; I+B+) ewes. The high OR ewes were heterozygous carriers of mutations in BMP15 (I+) and BMPRIB (B+). Functional differences in follicular somatic (granulosa and cumulus) cells between these genotypes, including differential gonadotropin responsiveness of granulosa cells, composition of follicular fluid and gene expression profiles in cumulus cells were evident. These differences emerged as part of a compensatory mechanism by which oocytes from smaller follicles, containing fewer granulosa cells, achieved developmental competency in I+B+ ewes.  The second objective of this PhD study was to develop new approaches for improving current in vitro maturation (IVM) strategies. The first approach utilised in this study focused on developing biomarkers that could be used to improve prediction of developmental competency in oocytes and in vitro produced embryos. This involved interrogating the hypothesis that a combination of molecular and morphokinetic biomarkers would better predict the developmental competency of oocytes and embryos compared to using these biomarkers alone. The second approach utilised in this PhD study tested the effects of modulating IVM conditions to better mimic the follicular microenvironment of a high, compared to a low, OR species on oocyte developmental competency and embryo quality. This involved supplementing IVM media with different ratios of two oocyte-secreted growth factors, i.e. GDF9:BMP15, that were representative of low or high OR species. These approaches demonstrated significant potential and warrant further investigation.  The most significant finding of this study was that despite variances in the surrounding microenvironment during in vivo and in vitro oocyte maturation that culminated in differential gene expression patterns in cumulus cells, and divergent gonadotropin-responsiveness of granulosa cells, the gene expression signatures of developmentally-competent oocytes and the morphokinetics of high-quality embryos were unaltered. This confirms the value of developing such biomarkers for oocyte development competency and embryo quality that remain unaltered despite a changing surrounding environment. Interestingly, simulating the ratio of GDF9:BMP15 that oocytes from high OR species are exposed to during maturation improved developmental competency in oocytes as demonstrated by increased blastocyst rates. Furthermore, this study has demonstrated that combinations of molecular (cumulus cell gene expression) and morphokinetic biomarkers improved the ability to predict developmental competency in oocytes and embryos. Overall, this study revealed novel information regarding the follicular microenvironment during final maturation and identified several novel approaches to improving the efficiency of ART.</p>

scholarly journals Piglets produced by transfer of embryos obtained by in vitro fertilization of oocytes matured in vitro with thymosin: A case report

2020 ◽  
Vol 76 (03) ◽  
pp. 6356-2020 ◽  
Author(s):  
KATARZYNA PONIEDZIAŁEK-KEMPNY ◽  
BARBARA GAJDA ◽  
IWONA RAJSKA ◽  
LECHOSŁAW GAJDA ◽  
ZDZISŁAW SMORĄG
Keyword(s):  
Case Report ◽  
In Vitro Fertilization ◽  
Control Group ◽  
First Birth ◽  
Vitro Fertilization ◽  
Research Material ◽  
Preliminary Study ◽  
Experimental Group

The aim of the study was to examine the in vivo viability of in vitro-produced (IVP) porcine embryos obtained from oocytes matured with thymosin. The research material for this study consisted of immature pig oocytes obtained from ovaries after slaughter and ejaculated semen obtained from one boar. The immature oocytes were cultured in vitro until the metaphase II stage in a medium supplemented with thymosin (TMS). The presumptive zygotes obtained were cultured in vitro for 4-40 hours. The presumptive zygotes and 2-4-cell embryos were evaluated in vivo after transferring them to synchronized recipients. After the transfer of embryos from the experimental group into 2 recipients (50 embryos into each gilt) and the transfer of 50 embryos from the control group into 1 recipient, both gilts that had received embryos obtained by in vitro fertilization of oocytes matured with TMS became pregnant and delivered a total of 16 live piglets. After the transfer of embryos from the control group, no pregnancy was achieved. In conclusion, the results of our preliminary study suggest that the maturation of pig oocytes with thymosin supports the in vivo survival of in vitro produced embryos. It is important to note, that this was the first birth of piglets obtained after transfer of IVP embryos in Poland.

scholarly journals Features of the preparation of biological material for genome editing in cattle

2019 ◽  
Vol 191 (12) ◽  
pp. 40-44
Author(s):  
A. Barkova ◽  
M. Modorov ◽  
G. Isaeva ◽  
A. Krivonogova
Keyword(s):  
Genome Editing ◽  
Biological Material ◽  
Genetic Material ◽  
Cumulus Cells ◽  
Vitro Fertilization ◽  
Expected Time ◽  
Donor Material ◽  
Material Transportation

Abstract. To carry out genome editing in cattle, an effective and well-functioning system for obtaining gametes, fertilizing eggs and their cryopreservation is necessary. Aim of the work: review and research of present-day existing methods of obtaining, insemination and cryopreservation of donor material, in order to provide genome editing in cows. Methods and materials. The work is completed according to the theme No. 0532-2019-0001 “Development of complex technology of marker-based genome selection of agricultural animals” within State Order of Ministry of Education and Science of the Russian Federation. The analysis of open scientific literature on the issues of in vitro fertilization in animals, cryopreservation of oocytes and embryons, sperm preparation and methods of insemination of cows’ oocytes, and cryopreservation of oocytes and embryons of animals is done. Features of the preparation of biological material of cattle for genome editing by microinjection into ooplasm are described. Results of research and duscussion. At present time there are two ways to obtain donor material from cattle: from live animals and taking ovaries after slaughtering cows. Material transportation is carried out at a temperature of 30–37 °C depending on the distance to the laboratory and expected time period of transportation. Oocyte-cumulus complexes can be removed by ovarian dissection and aspiration of visible follicles. In both cases, immature eggs are predominantly obtained. Subsequent ripening is carried out in vitro using special media in a CO2 incubator. The culture medium for oocyte maturation should contain hormones that mimic the peak of LH (luteinizing hormone), which occurs in vivo during the maturation of oocytes before ovulation. To accumulate a certain number of eggs at the stage of MII, it is recommended to carry out their cryopreservation by the method of vitrification, having previously released the oocyte from the cumulus cells. After thawing, oocytes need to be incubated for 2–3 hours 38.5 °C in 5–6.5% CO2 to restore the spindle. In order to make editing more effective, the introduction of genetic material is recommended to be carried out in parallel with the fertilization method “icsi”. In humans, mice and rabbits, an injection of sperm into the cytoplasm is sufficient to activate the oocyte, however, in cattle, just micro-injection of the sperm is not enough and often the male pronucleus does not form. To solve the problem, various methods are used, including freezing-thawing of sperm, resulting in damage of a membrane, or addition of heparin-glutathione into the medium that increases decondensation of the sperm DNA.

scholarly journals Clonal Micropropagation of Cranberry (Oxycoccus palustris Pers.)

2021 ◽  
Vol 51 (1) ◽  
pp. 67-76
Author(s):  
Sergey Makarov ◽  
Irina Kuznetsova ◽  
Mikhail Upadyshev ◽  
Sergey Rodin ◽  
Anton Chudetsky
Keyword(s):  
Survival Rate ◽  
Nutrient Medium ◽  
Average Length ◽  
Forest Products ◽  
Hybrid Form ◽  
Clonal Micropropagation ◽  
Planting Material ◽  
Total Growth

Introduction. The last decade saw a considerable increase in the demand for European cranberry planting material (Oxyccocus palustris Pers.) among consumers of non-timber forest products. Cranberry possesses high nutritional and medicinal value. Cultivars and hybrids of European cranberry prove extremely productive for plantation growth using the method of clonal micropropagation with revitalized planting material. Study objects and methods. The research featured European cranberry plants of the Dar Kostromy cultivar and its hybrid form 1-15-635. The study focused on the effect of various medications and growth regulators on the biometric profile of European cranberry and its adaptation to non-sterile conditions at all stages of in vivo clonal micropropagation. Results and discussion. During the introduction stage, the highest viability belonged to the explants treated with AgNO3 (95–96%) and Lizoformin 3000 (5%) as the main sterilizing solutions at a 10-min exposure and a 5% solution of Ecosterilizer (1:1) at a 20-min exposure (90–95%). During the micropropagation proper, the number, average length, and total growth of shoots increased as the concentration of cytokinin 2ip in the WPM 1/4 nutrient medium rose from 1.0 to 5.0 mg/L. At the stage of in vitro rooting, the maximal number, average length, and total growth of roots in regenerated plants for both cultivars were observed when Kornerost 5.0 mg/L was added to the WPM 1/4 nutrient medium. At the stage of adaptation to in vivo conditions, Micogel 0.2 mg/L contributed to the highest survival rate (94–100%). Conclusion. During clonal micropropagation in vitro, the biometric profile of European cranberry (Oxyccocus palustris Pers.) and its survival rate under non-sterile conditions in vivo proved to depend on various growth-regulating substances and their concentrations.

scholarly journals Radiosynthesis, in vitro and preliminary in vivo evaluation of the novel glutamine derived PET tracers [18F]fluorophenylglutamine and [18F]fluorobiphenylglutamine

2020 ◽  
Author(s):  
Tristan Baguet ◽  
Jeroen Verhoeven ◽  
Glenn Pauwelyn ◽  
Jiyun Hu ◽  
Patricia Lambe ◽  
...  
Keyword(s):  
Phenyl Group ◽  
Tumor Model ◽  
High Quality ◽  
In Vivo Evaluation ◽  
Pet Tracers ◽  
Imaging Tool ◽  
Starting Point ◽  
Inhibition Experiment

AbstractIntroductionGlucose has been deemed the driving force of tumor growth for decades. However, research has shown that several tumors metabolically shift towards glutaminolysis. The development of radiolabeled glutamine derivatives could be a useful molecular imaging tool for visualizing these tumors. We elaborated on the glutamine-derived PET tracers by developing two novel probes, namely [18F]fluorophenylglutamine and [18F]fluorobiphenylglutamineMaterials and methodsBoth tracers were labelled with fluorine-18 using our recently reported ruthenium-based direct aromatic fluorination method. Their affinity was evaluated with a [3H]glutamine inhibition experiment in a human PC-3 and a rat F98 cell line. The imaging potential of [18F]fluorophenylglutamine and [18F]fluorobiphenylglutamine was tested using a mouse PC-3 and a rat F98 tumor model.ResultsThe radiosynthesis of both tracers was successful with overall non-decay corrected yields of 18.46 ± 4.18 % (n=10) ([18F]fluorophenylglutamine) and 8.05 ± 3.25 % (n=5) ([18F]fluorobiphenylglutamine). In vitro inhibition experiments showed a moderate and low affinity of fluorophenylglutamine and fluorobiphenylglutamine, respectively, towards the human ASCT-2 transporter. Both compounds had a low affinity towards the rat ASCT-2 transporter. These results were endorsed by the in vivo experiments with low uptake of both tracers in the F98 rat xenograft, low uptake of [18F]FBPG in the mice PC-3 xenograft and a moderate uptake of [18F]FPG in the PC-3 tumors.ConclusionWe investigated the imaging potential of two novel PET radiotracers [18F]FPG and [18F]FBPG. [18F]FPG is the first example of a glutamine radiotracer derivatized with a phenyl group which enables the exploration of further derivatization of the phenyl group to increase the affinity and imaging qualities. We hypothesize that increasing the affinity of [18F]FPG by optimizing the substituents of the arene ring can result in a high-quality glutamine-based PET radiotracer.Advances in Knowledge and Implications for patient careWe hereby report novel glutamine-based PET-tracers. These tracers are tagged on the arene group with fluorine-18, hereby preventing in vivo defluorination, which can occur with alkyl labelled tracers (e.g. (2S,4R)4-[18F]fluoroglutamine). [18F]FPG shows clear tumor uptake in vivo, has no in vivo defluorination and has a straightforward production. We believe this tracer is a good starting point for the development of a high-quality tracer which is useful for the clinical visualization of the glutamine transport.

scholarly journals Sanitization of potato varieties from the VIR collection against viruses

2021 ◽  
Vol 181 (4) ◽  
pp. 164-172
Author(s):  
E. S. Bespalova ◽  
M. M. Agakhanov ◽  
S. B. Arkhimandritova ◽  
M. V. Erastenkova ◽  
Yu. V. Uhatova
Keyword(s):  
Solanum Tuberosum ◽  
Apical Meristem ◽  
Multiple Infections ◽  
Meristem Culture ◽  
Close Monitoring ◽  
High Quality ◽  
Potato Plants ◽  
Planting Material ◽  
Potato Collection

Background. VIR’s potato collection is one of the oldest and richest; however, it is constantly exposed to viruses that negatively affect useful agronomic properties of tubers. Close monitoring of the phytosanitary state of potato accessions helps to select the most effective method of therapy for subsequent healing of infected plants and obtaining high-quality planting material.Materials and methods. The research was aimed at improving the health of 18 varieties of Solanum tuberosum L. from the VIR collection. Testing for the presence of viruses was based on the ICA and RTPCR techniques, and the consequent healing was performed using the methods of meristem culture and cryotherapy.Results and conclusions. During the field test of potato plants, PVX, PVS and PVA were found to be the most common viruses. PSTVd was completely absent in all tested accessions. The effectiveness of in vitro healing of potato plants from viruses was assesses using meristem culture. The percentage of healed plants was 0% for PVS, 0% for PVX, 33.4% for PVA, 50% for PLRV, 72.3% for PVY, and 83.4% for PVM. Healing with meristem culture was shown to be the most effective against PVY and PVM. While assessing the effectiveness of post-cryogenic restoration of potato microplants, the level of post-cryogenic regeneration of the shoot tips in potato microplants was determined at 22.3% on average for a sample. The minimum was observed in k-16762 ‘Sagita N’ (5%), and the maximum in k-1378 ‘Marta’ (41.7%). Analysis of the effectiveness of potato recovery from viruses by in vitro cryotherapy showed that the percentage of recovered plants was 100% for PVY, 100% for PVA, 88.9% for PVM, 77.8% for PVS, 44.4% for PVX. Thus, the techniques of apical meristem culture and cryotherapy proved to be effective against PVY, PVA and PVM viruses. However, in the case of multiple infections, it is necessary to combine elements of different healing protocols to increase the effectiveness of the healing procedure. 

scholarly journals A PMMA-Based Microfluidic Device for Human Sperm Evaluation and Screening on Swimming Capability and Swimming Persistence

Micromachines ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 793
Author(s):  
Yimo Yan ◽  
Haoran Liu ◽  
Boxuan Zhang ◽  
Ran Liu
Keyword(s):  
Human Sperm ◽  
High Viscosity ◽  
Control Group ◽  
High Quality ◽  
Vitro Fertilization ◽  
Step Procedure ◽  
Motion Characteristics ◽  
Selection Of

The selection of high-quality sperm is essential to the success of in vitro fertilization (IVF). As human cervical mucus has a high viscosity, without enough swimming persistence, human sperm clouds cannot arrive at the ampulla to fertilize the egg. In this study, we used swimming capability and motion characteristics that are known to be associated with fertilization ability to evaluate the quality of sperm. Here, a clinically applicable polymethyl methacrylate (PMMA)-based microdevice was designed and fabricated for sperm evaluation and screening for swimming capability and persistence in a viscous environment. In this study, we applied methylcellulose (MC) to mimic the natural properties of mucus in vivo to achieve the selection of motile sperm. Sperm motion was recorded by an inverted microscope. The statistical features were extracted and analyzed. Hundreds of sperm in two treated groups with different concentrations of MC and one control group with human tubal fluid (HTF) media were video recorded. This device can achieve a one-step procedure of high-quality sperm selection and achieve the quantitative evaluation of sperm swimming capability and persistence. Sperm with good swimming capability and persistence may be more suitable for fertilization in a viscous environment. This microdevice and methods could be used to guide the evaluation of sperm motility and screening in the future.

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