scholarly journals Off-Stoichiometric Reactions at the Cell–Substrate Biomolecular Interface of Biomaterials: In Situ and Ex Situ Monitoring of Cell Proliferation, Differentiation, and Bone Tissue Formation

2019 ◽  
Vol 20 (17) ◽  
pp. 4080 ◽  
Author(s):  
Giuseppe Pezzotti ◽  
Tetsuya Adachi ◽  
Francesco Boschetto ◽  
Wenliang Zhu ◽  
Matteo Zanocco ◽  
...  
Keyword(s):  
Bone Formation ◽  
Cell Metabolism ◽  
Mesenchymal Cell ◽  
Ex Situ ◽  
Spectroscopic Techniques ◽  
Material Interface ◽  
Chemical Equations ◽  
Biomedical Titanium Alloy

The availability of osteoinductive biomaterials has encouraged new therapies in bone regeneration and has potentially triggered paradigmatic shifts in the development of new implants in orthopedics and dentistry. Among several available synthetic biomaterials, bioceramics have gained attention for their ability to induce mesenchymal cell differentiation and successive bone formation when implanted in the human body. However, there is currently a lack of understanding regarding the fundamental biochemical mechanisms by which these materials can induce bone formation. Phenomenological studies of retrievals have clarified the final effect of bone formation, but have left the chemical interactions at the cell–material interface uncharted. Accordingly, the knowledge of the intrinsic material properties relevant for osteoblastogenesis and osteoinduction remains incomplete. Here, we systematically monitored in vitro the chemistry of mesenchymal cell metabolism and the ionic exchanges during osteoblastogenesis on selected substrates through conventional biological assays as well as via in situ and ex situ spectroscopic techniques. Accordingly, the chemical behavior of different bioceramic substrates during their interactions with mesenchymal cells could be unfolded and compared with that of biomedical titanium alloy. Our goal was to clarify the cascade of chemical equations behind the biological processes that govern osteoblastogenic effects on different biomaterial substrates.

Collection and in vitro maturation of Mazama gouazoubira (brown brocket deer) oocytes obtained after ovarian stimulation

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Luciana Diniz Rola ◽  
Eveline dos Santos Zanetti ◽  
Maite del Collado ◽  
Ellen de Fátima Carvalho Peroni ◽  
José Maurício Barbanti Duarte
Keyword(s):  
Oocyte Maturation ◽  
Ovarian Stimulation ◽  
In Vitro Maturation ◽  
Wildlife Conservation ◽  
Estradiol Benzoate ◽  
Ex Situ ◽  
Follicular Aspiration ◽  
Mazama Gouazoubira

Summary In vitro production of embryos has gained prominence as a tool for use in wildlife conservation programmes in situ and ex situ. However, the development of this technique depends on steps that include ovarian stimulation, collection and oocyte maturation. The purpose of this study was to assess the feasibility of an ovarian stimulation protocol for follicular aspiration, the efficiency of videolaparoscopy for follicular aspiration and test a medium for in vitro oocyte maturation for the species Mazama gouazoubira. Five females were submitted to repeated ovarian stimulation (hormone protocol using controlled internal drug release), and estradiol benzoate on D0 and eight injections of follicle-stimulating hormone, once every 12 h, from D4 onwards at 30-day intervals. Fourteen surgical procedures were performed in superstimulated females, resulting in the collection of 94 oocytes and an average of 17.1 ± 9.1 follicles observed, 13.5 ± 6.6 follicles aspirated and 7.2 ± 3.7 oocytes collected per surgery. After collection, the oocytes were submitted to in vitro maturation for 24 h and stained with Hoechst 33342 dye to evaluate their nuclear status; 64.5% of the oocytes reached MII and 16.1% were spontaneously activated by parthenogenesis. The nuclear status of oocytes that did not undergo in vitro maturation was evaluated; 80.9% were found to be immature.

scholarly journals Ex situ conservation of genetic resources of field elm (Ulmus minor Mill) and European white elm (Ulmus laevis Pall)

Genetika ◽  
2004 ◽  
Vol 36 (3) ◽  
pp. 221-227
Author(s):  
Jelena Aleksic ◽  
Sasa Orlovic
Keyword(s):  
Genetic Resources ◽  
Ex Situ Conservation ◽  
Ex Situ ◽  
In Vitro Production ◽  
Ulmus Minor ◽  
Conservation Of Genetic Resources ◽  
Ulmus Laevis ◽  
Vitro Production

Principles of the conservation of genetic resources of elms (Ulmus spp) do not differ fundamentally from the general principles accepted for the conservation of genetic resources of other common Noble Hardwoods. Efficient conservation can best be achieved through appropriate combination of in situ and ex situ methods, which have distinct advantages. Besides that, ex situ conservation is employed when emergency measures are needed for rare endangered populations and when populations are too small to be managed in situ (e.g. risks of genetic drift and inbreeding). The aim of our research is ex situ conservation of genetic resources of field elm {Ulmus minor Mill) and European white elm (Ulmus laevis Pall) through establishment of field genebanks. Sampling was conducted in one population of field elm and one population of white elm. Plant material (buds) from 8 trees of field elm and 10 trees of white elm was used for in vitro production of clones. Obtained clones will be used for establishment of field genebanks on the experimental estate of the Institute of Lowland Forestry and Environment.

scholarly journals Application of the MSAP Technique to Evaluate Epigenetic Changes in Plant Conservation

2020 ◽  
Vol 21 (20) ◽  
pp. 7459
Author(s):  
María Elena González-Benito ◽  
Miguel Ángel Ibáñez ◽  
Michela Pirredda ◽  
Sara Mira ◽  
Carmen Martín
Keyword(s):  
Dna Methylation ◽  
In Situ Conservation ◽  
Plant Conservation ◽  
Ex Situ ◽  
Conservation Strategies ◽  
Regenerated Plants ◽  
Before And After ◽  
Methylation Patterns

Epigenetic variation, and particularly DNA methylation, is involved in plasticity and responses to changes in the environment. Conservation biology studies have focused on the measurement of this variation to establish demographic parameters, diversity levels and population structure to design the appropriate conservation strategies. However, in ex situ conservation approaches, the main objective is to guarantee the characteristics of the conserved material (phenotype and epi-genetic). We review the use of the Methylation Sensitive Amplified Polymorphism (MSAP) technique to detect changes in the DNA methylation patterns of plant material conserved by the main ex situ plant conservation methods: seed banks, in vitro slow growth and cryopreservation. Comparison of DNA methylation patterns before and after conservation is a useful tool to check the fidelity of the regenerated plants, and, at the same time, may be related with other genetic variations that might appear during the conservation process (i.e., somaclonal variation). Analyses of MSAP profiles can be useful in the management of ex situ plant conservation but differs in the approach used in the in situ conservation. Likewise, an easy-to-use methodology is necessary for a rapid interpretation of data, in order to be readily implemented by conservation managers.

scholarly journals Tetrathiafulvalene: A Gate to the Mechanochemical Mechanisms of Electron Transfer Reactions

Crystals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 482
Author(s):  
Richard Chen ◽  
Mehmet Kerem Gokus ◽  
Silvina Pagola
Keyword(s):  
Mechanochemical Synthesis ◽  
Amorphous Material ◽  
In Situ Monitoring ◽  
Ex Situ ◽  
Mechanochemical Reaction ◽  
Charge Transfer Complexes ◽  
Spectroscopic Techniques ◽  
Diffusion Controlled ◽  
Mechanochemical Reactions

This report describes aspects of our previous studies of the mechanochemical synthesis of charge transfer complexes of the electron donor tetrathiafulvalene, which are relevant to the use of laboratory X-ray powder diffraction for ex situ monitoring of mechanochemical reactions toward investigating their mechanisms. In particular, the reaction of tetrathiafulvalene and chloranil was studied under neat mechanochemical conditions and liquid-assisted grinding with diethyl ether (1 μL/mg). The product in both cases is the green tetrathiafulvalene chloranil polymorph and the mechanism of the redox reaction is presumably the same. However, while the kinetic profile of the neat mechanochemical synthesis was fitted with a second-order rate law, that of the overall faster liquid-assisted grinding reaction was fitted with the Ginstling-Brounshtein 3D diffusion-controlled model. Hence, the diffusional processes and mass transfer bringing the reactants together and separating them from products must be different. Diffraction measurements sensitive to crystalline phases and amorphous material, combined with in situ monitoring by spectroscopic techniques, will ultimately afford a better understanding of mechanochemical reaction mechanisms, a hot topic in mechanochemistry.

scholarly journals Combined in situ Physical and ex-situ Biochemical Approaches to Investigate in vitro Deconstruction of Destarched Wheat Bran by Enzymes Cocktail Used in Animal Nutrition

2019 ◽  
Vol 7 ◽  
Author(s):  
Marine Deshors ◽  
Olivier Guais ◽  
Virginie Neugnot-Roux ◽  
Xavier Cameleyre ◽  
Luc Fillaudeau ◽  
...  
Keyword(s):  
Wheat Bran ◽  
Ex Situ ◽  
Animal Nutrition

NMR methods for in-situ biofilm metabolism studies: spatial and temporal resolved measurements

2005 ◽  
Vol 52 (7) ◽  
pp. 7-12 ◽  
Author(s):  
P.D. Majors ◽  
J.S. McLean ◽  
J.K. Fredrickson ◽  
R.A. Wind
Keyword(s):  
Magnetic Resonance ◽  
Cell Metabolism ◽  
Shewanella Oneidensis ◽  
Growth Conditions ◽  
Magnetic Resonance Images ◽  
Ex Situ ◽  
Adherent Cell ◽  
Growth Environment ◽  
Environment Control

We are developing novel nuclear magnetic resonance (NMR) microscopy, spectroscopy and combined NMR/optical techniques for the study of biofilms under known, controlled growth conditions. Objectives include: time and depth-resolved metabolite concentrations with isotropic spatial resolution on the order of 10 microns, metabolic pathways and flux rates, mass transport and ultimately their correlation with gene expression by optical microscopy in biofilms. We describe the implementation of ex-situ grown biofilms to improve growth environment control and NMR analysis. In-situ NMR depth resolved metabolite profiling techniques are introduced and demonstrated for a Shewanella oneidensis strain MR-1 biofilm. Finally, initial combined confocal fluorescence and magnetic resonance images are shown for a GFP-labeled Shewanella biofilm. These methods are equally applicable to other biofilm systems of interest; thus they may provide a significant contribution toward the understanding of adherent cell metabolism.

scholarly journals Long-term conservation of potato genetic resources: Methods and status of conservation

2019 ◽  
pp. 717-725
Author(s):  
Jane Muthoni ◽  
Hussein Shimelis ◽  
Rob Melis
Keyword(s):  
Genetic Resources ◽  
Slow Growth ◽  
Plant Genetic Resources ◽  
Ex Situ ◽  
Growth Media ◽  
Medium Term ◽  
Natural Habitats

Plant genetic resources (PGRs) play an important role in agriculture, environment protection, cultural property and trade; they need to be conserved. There are two fundamental approaches for the conservation of PGRs: in situ and ex situ. In situ conservation is the conservation of ecosystems and natural habitats and the maintenance and recovery of viable populations of species in their natural surroundings. Ex situ preservation is the storage of seeds or plant materials under artificial conditions to maintain their long term viability and availability for use. Genebanks employ seed storage, field collections of living plants and in vitro storage (tissue culture or cryopreservation) for ex situ preservation of PGR. Storage of orthodox seeds, which are tolerant to low moisture content and low temperatures at appropriate temperature and humidity, is the most convenient ex situ conservation method. Plants that produce recalcitrant seeds or non-viable seeds are conserved in field genebanks as well as in-vitro in slow growth media for short-to-medium term and cryopreservation in liquid nitrogen at -1960C for long-term periods. Cryopreservation is very expensive and needs trained personnel; this could explain why this method is rarely used for conservation of plant genetic resources in most developing countries. Potato tubers are bulky and highly perishable; the crop is generally conserved as clones either in field genebanks (with annual replanting), in-vitro conservation in slow growth media for short-to-medium term and cryopreservation for long term. Field genebanks are expensive to maintain and the crop is exposed to many dangers; hence, cryopreservation is the only feasible method for long term conservation. However, given the high cost of cryopreservation, long-term conservation of potato genetic resources is poorly developed in most resource-poor countries leading to high rates of genetic erosion. This paper looks into the various methods that that can be applied to conserve potato genetic resources and the status of conservation of potatoes in major genebanks and some countries.

Applications of Synchrotron Surface X-Ray Scattering Studies of Electrochemical Interfaces

MRS Bulletin ◽  
1999 ◽  
Vol 24 (1) ◽  
pp. 36-40 ◽  
Author(s):  
Hoydoo You ◽  
Zoltán Nagy
Keyword(s):  
Surface Science ◽  
Region Of Interest ◽  
Atomic Layer ◽  
Living Environment ◽  
Ex Situ ◽  
Spectroscopic Techniques ◽  
Engineering Research ◽  
Significant Intensity ◽  
Electrochemical Interfaces

Aqueous-solution/solid interfaces are ubiquitous in modern manufacturing environments as well as in our living environment, and studies of such interfaces are an active area of science and engineering research. An important area is the study of liquid/solid interfaces under active electrochemical control, which has many immediate technological implications, for example, corrosion/passivation of metals and energy storage in batteries and ultracapacitors. The central phenomenon of electrochemistry is the charge transfer at the interface, and the region of interest is usually wider than a single atomic layer, ranging from a monolayer to thousands of angstroms, extending into both phases.Despite the technological and environmental importance of liquid/solid interfaces, the atomic level understanding of such interfaces had been very much hampered by the absence of nondestructive, in situ experimental techniques. The situation has changed somewhat in recent decades with the development of the largely ex situ ultrahigh vacuum (UHV) surface science, modern spectroscopic techniques, and modern surface microscopy.However in situ experiments of electrochemical interfaces are difficult, stemming from the special nature of these interfaces. These are so-called buried interfaces in which the solid electrode surface is covered by a relatively thick liquid layer. For this reason, the probe we use in the structural investigation must satisfy simultaneously two conditions: (1) the technique must be surface/interface sensitive, and (2) absorption of the probe in the liquid phase must be sufficiently small for penetration to and from the interface of interest without significant intensity loss.

scholarly journals Preserving grapevine variety Fioletoviy Ranniy in the collection in vitro

2021 ◽  
Vol 273 ◽  
pp. 01007
Author(s):  
Valentina Puzirnova ◽  
Natalia Doroshenko
Keyword(s):  
Biodiversity Conservation ◽  
Sustainable Management ◽  
Future Generations ◽  
Ex Situ ◽  
Grapevine Variety ◽  
The World ◽  
History Of ◽  
Slow Growing

The paper is devoted to the problem of plant biodiversity conservation. This problem is acute all over the world. Lower Don Region has a centuries old history of viticulture and winemaking. There are many valuable vine varieties which are worthy of preservation for future generations. Classical methods no longer cope with this task. Applying advances of biotechnology in addition to traditional methods of ex situ and in situ biodiversity conservation allows sustainable management of genetic resources. This article summarizes the study of methods for creation slow growing collection for grapevine variety Fioletoviy Ranniy. Keeping plants in a slow-growing collection is one of the best ways to preserve biodiversity. This study analyzed the effect of various media compounds on vigor of vine in order to elongate the time between replantings.

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