Vitrification in human and domestic animal embryology: work in progress

2013 ◽  
Vol 25 (5) ◽  
pp. 719 ◽  
Author(s):  
Gábor Vajta

According to the analysis of papers published in major international journals, rapidly increasing application of vitrification is one of the greatest achievements in domestic animal and especially human embryology during the first decade of our century. This review highlights factors supporting or hampering this progress, summarises results achieved with vitrification and outlines future tasks to fully exploit the benefits of this amazing approach that has changed or will change many aspects of laboratory (and also clinical) embryology. Supporting factors include the simplicity, cost efficiency and convincing success of vitrification compared with other approaches in all species and developmental stages in mammalian embryology, while causes that slow down the progress are mostly of human origin: inadequate tools and solutions, superficial teaching, improper application and unjustified concerns resulting in legal restrictions. Elimination of these hindrances seems to be a slower process and more demanding task than meeting the biological challenge. A key element of future progress will be to pass the pioneer age, establish a consensus regarding biosafety requirements, outline the indispensable features of a standard approach and design fully-automated vitrification machines executing all phases of the procedure, including equilibration, cooling, warming and dilution steps.

2019 ◽  
Author(s):  
Wenqing Cai ◽  
Jialiang Huang ◽  
Qian Zhu ◽  
Bin E. Li ◽  
Davide Seruggia ◽  
...  

SummaryHow overall principles of gene regulation (the “logic”) may change during ontogeny is largely unexplored. We compared transcriptomic, epigenomic and topological profiles in embryonic (EryP) and adult (EryD) erythroblasts. Despite reduced chromatin accessibility compared to EryP, distal chromatin of EryD is enriched in H3K27ac, Gata1 and Myb occupancy. In contrast to EryP-specific genes, which exhibit promoter-centric regulation through Gata1, EryD-specific genes employ distal enhancers for long-range regulation through enhancer-promoter looping, confirmed by Gata1 HiChIP. Genome editing demonstrated distal enhancers are required for gene expression in EryD but not in EryP. Applying a metric for enhancer-dependence of transcription, we observed a progressive reliance on enhancer control with increasing age of ontogeny among diverse primary cells and tissues of mouse and human origin. Our findings highlight fundamental and conserved differences in regulatory logic at distinct developmental stages, characterized by simpler promoter-centric regulation in embryonic cells and combinatorial enhancer-driven control in adult cells.HighlightsRegulation of embryonic-specific erythroid genes is promoter-centric through Gata1Adult-specific control is combinatorial enhancer-driven and requires MybAdult specific genes have increased enhancer-promoter chromatin interactionsEnhancer-dependence increases progressively with increasing developmental age


Author(s):  
Tarana Siddika ◽  
Ilka U. Heinemann

MiRNAs are small non-coding RNAs that interact with their target mRNAs for posttranscriptional gene regulation. Finely controlled miRNA biogenesis, target recognition and degradation indicate that maintaining miRNA homeostasis is essential for regulating cell proliferation, growth, differentiation and apoptosis. Increasingly, miRNAs have been recognized as a potential biomarker for disease diagnosis. MiRNAs can be found in blood, plasma, and tissues, and miRNA expression and activity differ in developmental stages, tissues and in response to external stimuli. MiRNA transcripts are matured from pri-miRNA over pre-miRNA to mature miRNA, a process that includes multiple steps and enzymes. Many tools are available to identify and quantify specific miRNAs, ranging from measuring total miRNA, specific miRNA activity, miRNA arrays and miRNA localization. The various miRNA assays differ in accuracy, cost, efficiency and convenience of monitoring miRNA dynamics. To acknowledge the significance and increasing research interest in miRNAs, we summarize the traditional as well as novel methods of miRNA quantification with strengths and limitations of various techniques in biochemical and medical research.


Author(s):  
Simon Grant ◽  
Rowin Young

This paper reviews terminology, motivation, history and current work in areas relating to skill or competence. Many useful services, clarifying pathways within and from education to employment, self-assessment, and selection would be facilitated by better standardization of the format in which related definitions are represented, and also by a standard approach to representing the structured sets often called frameworks. To be effective, information models underlying interoperability specifications must be based on common conceptual models; the authors propose one such model as a work in progress. The authors see the way forward as reaching greater consensus about the components of competence, including intended learning outcomes, agreement on a model for frameworks allowing reuse of and comparison between components in and between frameworks, and investigation of how requirements and claims for skill and competence can be coordinated in the light of common practice in recruitment.


Author(s):  
Simon Grant ◽  
Rowin Young

This paper reviews terminology, motivation, history and current work in areas relating to skill or competence. Many useful services, clarifying pathways within and from education to employment, self-assessment, and selection would be facilitated by better standardization of the format in which related definitions are represented, and also by a standard approach to representing the structured sets often called frameworks. To be effective, information models underlying interoperability specifications must be based on common conceptual models; the authors propose one such model as a work in progress. The authors see the way forward as reaching greater consensus about the components of competence, including intended learning outcomes, agreement on a model for frameworks allowing reuse of and comparison between components in and between frameworks, and investigation of how requirements and claims for skill and competence can be coordinated in the light of common practice in recruitment.


1976 ◽  
Vol 32 ◽  
pp. 109-116 ◽  
Author(s):  
S. Vauclair

This paper gives the first results of a work in progress, in collaboration with G. Michaud and G. Vauclair. It is a first attempt to compute the effects of meridional circulation and turbulence on diffusion processes in stellar envelopes. Computations have been made for a 2 Mʘstar, which lies in the Am - δ Scuti region of the HR diagram.Let us recall that in Am stars diffusion cannot occur between the two outer convection zones, contrary to what was assumed by Watson (1970, 1971) and Smith (1971), since they are linked by overshooting (Latour, 1972; Toomre et al., 1975). But diffusion may occur at the bottom of the second convection zone. According to Vauclair et al. (1974), the second convection zone, due to He II ionization, disappears after a time equal to the helium diffusion time, and then diffusion may happen at the bottom of the first convection zone, so that the arguments by Watson and Smith are preserved.


Author(s):  
J. P. Revel

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.


Author(s):  
J. R. Adams ◽  
G. J Tompkins ◽  
A. M. Heimpel ◽  
E. Dougherty

As part of a continual search for potential pathogens of insects for use in biological control or on an integrated pest management program, two bacilliform virus-like particles (VLP) of similar morphology have been found in the Mexican bean beetle Epilachna varivestis Mulsant and the house cricket, Acheta domesticus (L. ).Tissues of diseased larvae and adults of E. varivestis and all developmental stages of A. domesticus were fixed according to procedures previously described. While the bean beetles displayed no external symptoms, the diseased crickets displayed a twitching and shaking of the metathoracic legs and a lowered rate of activity.Examinations of larvae and adult Mexican bean beetles collected in the field in 1976 and 1977 in Maryland and field collected specimens brought into the lab in the fall and reared through several generations revealed that specimens from each collection contained vesicles in the cytoplasm of the midgut filled with hundreds of these VLP's which were enveloped and measured approximately 16-25 nm x 55-110 nm, the shorter VLP's generally having the greater width (Fig. 1).


Author(s):  
Regina Birchem

Spheroids of the green colonial alga Volvox consist of biflagellate Chlamydomonad-like cells embedded in a transparent sheath. The sheath, important as a substance through which metabolic materials, light, and the sexual inducer must pass to and from the cells, has been shown to have an ordered structure (1,2). It is composed of both protein and carbohydrate (3); studies of V. rousseletii indicate an outside layer of sulfated polysaccharides (4).Ultrastructural studies of the sheath material in developmental stages of V. carteri f. weismannia were undertaken employing variations in the standard fixation procedure, ruthenium red, diaminobenzidine, and high voltage electron microscopy. Sheath formation begins after the completion of cell division and inversion of the daughter spheroids. Golgi, rough ER, and plasma membrane are actively involved in phases of sheath synthesis (Fig. 1). Six layers of ultrastructurally differentiated sheath material have been identified.


Author(s):  
Y. R. Chen ◽  
Y. F. Huang ◽  
W. S. Chen

Acid phosphatases are widely distributed in different tisssues of various plants. Studies on subcellular localization of acid phosphatases show they might be present in cell wall, plasma lemma, mitochondria, plastid, vacuole and nucleus. However, their localization in rice cell varies with developmental stages of cells and plant tissues. In present study, acid phosphatases occurring in root cap are examined.Sliced root tips of ten-day-old rice(Oryza sativa) seedlings were fixed in 0.1M cacodylate buffer containing 2.5% glutaraldehyde for 2h, washed overnight in same buffer solution, incubated in Gomori's solution at 37° C for 90min, post-fixed in OsO4, dehydrated in ethanol series and finally embeded in Spurr's resin. Sections were doubly stained with uranyl acetate and lead citrate, and observed under Hitachi H-600 at 75 KV.


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