The Culture of Small Aggregates of Amphibian Embryonic Cells in vitro

Development ◽  
1963 ◽  
Vol 11 (1) ◽  
pp. 135-154
Author(s):  
K. W. Jones ◽  
T. R. Elsdale
Keyword(s):  
Pigment Cells ◽  
Two Dimensional ◽  
Embryonic Cells ◽  
Common Procedure ◽  
Large Numbers ◽  
Organ Type ◽  
Typical Cell ◽  
Primary Explant ◽  
Migration Of Cells

A Common procedure in amphibian embryology has been to remove portions from embryos and culture these under conditions in which the large numbers of cells retain a close-knit association, favourable to the differentiation of primitive organs in the explant. It has not, in general, been the aim to employ the primary explant as a source of a two-dimensional outgrowth of cells on the substrate, as in typical cell culture procedures. Because of their inherent migratory tendencies, however, outgrowths of pigment cells are readily obtained from explants of the amphibian neural crest, and these have stimulated the interest of a number of investigators (see Wilde, 1961). Holtfreter (1938, 1946) and Finnegan (1953) have also observed the migration of cells from explants of Urodele embryos. Several investigators have employed cell cultures as opposed to organ type cultures in induction studies, Niu & Twitty (1953), Niu (1958), Barth & Barth (1959) and Becker, Tiedemann & Tiedemann (1959).

scholarly journals Protein changes during the asexual cycle of Neurospora crassa.

1985 ◽  
Vol 5 (4) ◽  
pp. 839-848 ◽  
Author(s):  
V Berlin ◽  
C Yanofsky
Keyword(s):  
Two Dimensional ◽  
Protein Profiles ◽  
Wild Type ◽  
Two Dimensional Gel Electrophoresis ◽  
Large Numbers ◽  
Aerial Hyphae ◽  
Reticulocyte Lysate ◽  
Steady State Levels ◽  
Asexual Cycle

A method for synchronizing conidiation and isolating large numbers of cells at discrete stages of conidia development is described. Using two-dimensional gel electrophoresis, we analyzed the protein profiles of mycelia, aerial hyphae, and conidia and observed that the concentration of 14 polypeptides increase and 38 decrease during the asexual cycle. Twelve polypeptides were present in extracts of aerial hyphae or conidia, but not mycelia, suggesting that they may be conidiation specific. The protein profiles of mutants defective in conidiation were also analyzed. Differences were detected in the two-dimensional profiles of protein extracts from fluffy and wild-type aerial hyphae. Polyadenylated RNA isolated from wild-type mycelia and conidiating cultures was translated in vitro in a rabbit reticulocyte lysate. Differences were detected in the polypeptide products specified by the two RNA populations, suggesting that changes in steady-state levels of polyadenylated RNAs also occur during conidiation.

Protein changes during the asexual cycle of Neurospora crassa

1985 ◽  
Vol 5 (4) ◽  
pp. 839-848
Author(s):  
V Berlin ◽  
C Yanofsky
Keyword(s):  
Two Dimensional ◽  
Protein Profiles ◽  
Wild Type ◽  
Two Dimensional Gel Electrophoresis ◽  
Large Numbers ◽  
Aerial Hyphae ◽  
Reticulocyte Lysate ◽  
Steady State Levels ◽  
Asexual Cycle

A method for synchronizing conidiation and isolating large numbers of cells at discrete stages of conidia development is described. Using two-dimensional gel electrophoresis, we analyzed the protein profiles of mycelia, aerial hyphae, and conidia and observed that the concentration of 14 polypeptides increase and 38 decrease during the asexual cycle. Twelve polypeptides were present in extracts of aerial hyphae or conidia, but not mycelia, suggesting that they may be conidiation specific. The protein profiles of mutants defective in conidiation were also analyzed. Differences were detected in the two-dimensional profiles of protein extracts from fluffy and wild-type aerial hyphae. Polyadenylated RNA isolated from wild-type mycelia and conidiating cultures was translated in vitro in a rabbit reticulocyte lysate. Differences were detected in the polypeptide products specified by the two RNA populations, suggesting that changes in steady-state levels of polyadenylated RNAs also occur during conidiation.

Cytoplasmic Granules in Lymphocytes from Rats Dosed with SK&F 14336-D

1971 ◽  
Vol 29 ◽  
pp. 556-557
Author(s):  
T. G. Merrill ◽  
B. J. Payne ◽  
A. J. Tousimis
Keyword(s):  
Lipid Storage ◽  
Pokeweed Mitogen ◽  
Electron Microscopic ◽  
Cytoplasmic Granules ◽  
Storage Diseases ◽  
Tay Sachs Disease ◽  
Large Numbers ◽  
Microscopic Studies ◽  
Neurovisceral Lipidosis

Rats given SK&F 14336-D (9-[3-Dimethylamino propyl]-2-chloroacridane), a tranquilizing drug, developed an increased number of vacuolated lymphocytes as observed by light microscopy. Vacuoles in peripheral blood of rats and humans apparently are rare and are not usually reported in differential counts. Transforming agents such as phytohemagglutinin and pokeweed mitogen induce similar vacuoles in in vitro cultures of lymphocytes. These vacuoles have also been reported in some of the lipid-storage diseases of humans such as amaurotic familial idiocy, familial neurovisceral lipidosis, lipomucopolysaccharidosis and sphingomyelinosis. Electron microscopic studies of Tay-Sachs' disease and of chloroquine treated swine have demonstrated large numbers of “membranous cytoplasmic granules” in the cytoplasm of neurons, in addition to lymphocytes. The present study was undertaken with the purpose of characterizing the membranous inclusions and developing an experimental animal model which may be used for the study of lipid storage diseases.

Specific Labeling of Protein Domains with Antibody Fragments

1981 ◽  
Vol 39 ◽  
pp. 416-417
Author(s):  
U. Aebi ◽  
L.E. Buhle ◽  
W.E. Fowler
Keyword(s):  
Image Processing ◽  
Specific Protein ◽  
Antibody Fragments ◽  
Supramolecular Organization ◽  
Two Dimensional ◽  
Ordered Structures ◽  
Virus Capsids ◽  
Processing Techniques

Many important supramolecular structures such as filaments, microtubules, virus capsids and certain membrane proteins and bacterial cell walls exist as ordered polymers or two-dimensional crystalline arrays in vivo. In several instances it has been possible to induce soluble proteins to form ordered polymers or two-dimensional crystalline arrays in vitro. In both cases a combination of electron microscopy of negatively stained specimens with analog or digital image processing techniques has proven extremely useful for elucidating the molecular and supramolecular organization of the constituent proteins. However from the reconstructed stain exclusion patterns it is often difficult to identify distinct stain excluding regions with specific protein subunits. To this end it has been demonstrated that in some cases this ambiguity can be resolved by a combination of stoichiometric labeling of the ordered structures with subunit-specific antibody fragments (e.g. Fab) and image processing of the electron micrographs recorded from labeled and unlabeled structures.

Biomechanical Evaluation of a New Fixation Method for Stabilization of Sagittal Split Ramus Osteotomy After Mandibular Advancement

2021 ◽  
Vol 6 ◽  
pp. 247275122110192
Author(s):  
Karel Kuik ◽  
Jean Pierre T. F. Ho ◽  
Cornelis Klop ◽  
Maurits H. T. de Ruiter ◽  
Cornelis J. Kleverlaan ◽  
...  
Keyword(s):  
Hybrid Method ◽  
In Vitro Study ◽  
Mandibular Advancement ◽  
Fixation Method ◽  
Common Procedure ◽  
Sagittal Split Ramus Osteotomy ◽  
Fixation Methods ◽  
Biomechanical Evaluation ◽  
Monocortical Screws

Study Design: Biomechanical in vitro study. Mandibular advancement after sagittal split ramus osteotomy (SSRO) is a common procedure in orthognathic surgery. Several fixation methods are used for stabilization of SSRO. Objective: The aim of this study was to compare a new fixation method (gridplate) with more contemporary applied methods of fixation. Material and Methods: In this study, 50 polyurethane hemimandibles with a prefabricated SSRO were used as specimens. All hemimandibles were advanced by 8 mm and divided into 5 groups with different fixation methods: (A) one 4-hole miniplate with 4 monocortical screws; (B) two 4-hole miniplates with 8 monocortical screws; (C) one 4-hole miniplate with 4 monocortical screws and 1 bicortical screw in the retromolar area (hybrid method); (D) 3 bicortical screws in an inverted-L pattern; and (E) one 8-hole gridplate with 8 monocortical screws. Loads in newtons were recorded at displacements of the mandibular incisive edge at 1 mm, 3 mm and 5 mm. Results: Fixation with 3 bicortical screws and the gridplate presented the most stability, followed by two 4-hole miniplates. Fixation with the hybrid method or the single miniplate showed the least stability. Conclusion: According to the results of this study, the 8-hole gridplate design appears a sufficient fixation method regarding stabilization of SSRO with larger mandibular advancement.

Animal Models and Alternatives in the Quality Control of Vaccines: Are In Vitro Methods or In Vivo Methods the Scientific Equivalent of the Emperor's New Clothes?

1995 ◽  
Vol 23 (1) ◽  
pp. 61-73
Author(s):  
Coenraad Hendriksen ◽  
Johan van der Gun
Keyword(s):  
Quality Control ◽  
Test Methods ◽  
In Vitro Test ◽  
Large Numbers ◽  
Alternative Approach ◽  
Inactivated Vaccines ◽  
Vitro Test

In the quality control of vaccine batches, the potency testing of inactivated vaccines is one of the areas requiring very large numbers of animals, which usually suffer significant distress as a result of the experimental procedures employed. This article deals with the potency testing of diphtheria and tetanus toxoids, two vaccines which are used extensively throughout the world. The relevance of the potency test prescribed by the European Pharmacopoeia monographs is questioned. The validity of the potency test as a model for the human response, the ability of the test to be standardised, and the relevance of the test in relation to the quality of the product are discussed. It is concluded that the potency test has only limited predictive value for the antitoxin responses to be expected in recipients of these toxoids. An alternative approach for estimating the potency of toxoid batches is discussed, in which a distinction is made between estimation of the immunogenic potency of the first few batches obtained from a seed lot and monitoring the consistency of the quality of subsequent batches. The use of animals is limited to the first few batches. Monitoring the consistency of the quality of subsequent batches is based on in vitro test methods. Factors which hamper the introduction and acceptance of the alternative approach are considered. Finally, proposals are made for replacement, reduction and/or refinement (the Three Rs) in the use of animals in the routine potency testing of toxoids.

Activation of the peptidergic neurosecretory system in Diphyllobothrium dendriticum (Cestoda: Pseudophyllidea)

Parasitology ◽  
1981 ◽  
Vol 83 (2) ◽  
pp. 243-247 ◽  
Author(s):  
Margaretha K. S. Gustafsson ◽  
Marianne C. Wikgren
Keyword(s):  
Neurosecretory Cells ◽  
Final Host ◽  
Fish Host ◽  
Neurosecretory System ◽  
Weak Reaction ◽  
Diphyllobothrium Dendriticum ◽  
Large Numbers ◽  
Short Times

SUMMARYThe activation of the peptidergic neurosecretory system in Diphyllobothrium dendriticum was studied following cultivation of plerocercoids for short times in vitro and in vivo. In the plerocercoid the neurosecretory cells gave a very weak reaction with paraldehyde fuchsin (PAF). After cultivation for 1 h large numbers of neurosecretory cells filled with PAF-positive granules were evident. The significance of the activation of the neurosecretory system during the transfer of the worm from the cold-blooded fish host to the warm-blooded final host is discussed.

scholarly journals Studies on the autophosphorylation of the insulin receptor from human placenta. Analysis of the sites phosphorylated by two-dimensional peptide mapping

1988 ◽  
Vol 252 (2) ◽  
pp. 607-615 ◽  
Author(s):  
J M Tavaré ◽  
R M Denton
Keyword(s):  
Thin Layer ◽  
Insulin Receptor ◽  
Peptide Mapping ◽  
Beta Subunit ◽  
Two Dimensional ◽  
Intact Cells ◽  
Tyrosine Residues ◽  
Domain 3 ◽  
Receptor Beta

1. A partially purified preparation of human placental insulin receptors was incubated with [gamma-32P]ATP in the presence or absence of insulin. The 32P-labelled insulin-receptor beta-subunits were then isolated, cleaved with trypsin followed by protease V8 and the [32P]phosphopeptides generated were analysed by thin layer electrophoresis and chromatography. This approach revealed that insulin stimulates autophosphorylation of the insulin-receptor beta-subunit in vitro on at least seven tyrosine residues distributed among three distinct domains. 2. One domain (domain 2), containing tyrosine residues 1146, 1150 and 1151 was the most rapidly phosphorylated and could be recovered as mono-, di- and triphosphorylated peptides cleaved by trypsin at Arg-1143 and either Lys-1153 or Lys-1156. Multiple phosphorylation of this domain appears to partially inhibit the cleavage at Lys-1153 by trypsin. 3. In a second domain (domain 3) containing two phosphorylated tyrosine residues at positions 1316 and 1322 the tyrosines were phosphorylated more slowly than those in domain 2. This domain is close to the C-terminus of the beta-subunit polypeptide chain. 4. At least two further tyrosine residues appeared to be phosphorylated after those in domains 2 and 3. These residues probably residue within a domain lying in close proximity to the inner face of the plasma membrane containing tyrosines 953, 960 and 972, but conclusive evidence is still required. 5. The two-dimensional thin-layer analysis employed in this study to investigate insulin-receptor phosphorylation has several advantages over previous methods based on reverse-phase chromatography. It allows greater resolution of 32P-labelled tryptic peptides and, when coupled to radioautography, is considerably more sensitive. The approach can be readily adapted to study phosphorylation of the insulin receptor within intact cells.

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