scholarly journals ADAPTATION DYNAMICS OF THE RABIES VIRUS OF "RV-97" VACCINE STRAIN TO MONOLAYER VNK-21/13 CELL CULTURE

2021 ◽  
Vol 250 ◽  
pp. 157-164
Author(s):  
A. V. Shishkov ◽  
◽  
A. A. Pyatkina ◽  
B. L. Manin ◽  
◽  
...  
Keyword(s):  
Cell Culture ◽  
Rabies Virus ◽  
Cell Monolayer ◽  
Test System ◽  
Fluorescent Label ◽  
Main Role ◽  
Phenotypic Properties ◽  
Passage Level ◽  
Almost All ◽  
Finite Cell

The problem of rabies as one of the most dangerous zoonoses continues to be relevant almost all over the world. In development of a live vaccine, an important stage is to obtain an active component - a virus that retains the given phenotypic properties, pathogen cultivation system plays the main role. The aim of this work was to adapt the rabies virus of “RV-97” strain to the finite cell line of the Syrian hamster kidney (VNK -21/13) of the Swedish subline, as well as to carry out a comparative analysis of virus accumulation at different passages. The number of passages that need to be carried out for adaptation of RV-97 strain to the monolayer culture of VNK -21/13 cells was determined. We used a 2-day culture of VNK -21/13 cells in the phase of logarithmic growth (80-90% formation of a cell monolayer). VNK -21/13 cell culture grown as a monolayer in the wells of flat-bottomed plastic plates was used as a test system for infectious activity. A fluorescent label was used to indicate infected cells. It was determined that the smallest number of passages at which the rabies virus of “RV-97” strain is adapted to the finite cell culture of VNK -21/13 of the Swedish subline is the 6th passage level. It was found that the titer of infectious activity of attenuated rabies virus of “RV-97” strain at the 6th passage level is 7.33 ± 0.17 lg CCID50 / cm3.

IN VITRO METHODS FOR THE STUDY OF THE ADENOHYPOPHYSIAL FUNCTIONS TO SECRETE GONADOTROPHIN

1971 ◽  
Vol 68 (1_Suppl) ◽  
pp. S27-S40 ◽  
Author(s):  
T. Kobayashi ◽  
T. Kigawa ◽  
M. Mizuno ◽  
T. Watanabe
Keyword(s):  
Cell Culture ◽  
Organ Culture ◽  
Cultured Cells ◽  
Cell Sheet ◽  
Short Term ◽  
Hypothalamic Extract ◽  
Numerous Cell ◽  
Single Cell Culture ◽  
Almost All

ABSTRACT There are several in vitro methods to analyse the function of the adenohypophysis or the mechanisms of its regulation. The present paper deals with single cell culture, organ culture and short term incubation techniques by which the morphology and gonadotrophin-secreting function of the adenohypophysis were studied. In trypsin-dispersed cell culture, the adenohypophysial cells showed extensive propagation to form numerous cell colonies and finally develop into a confluent monolayer cell sheet covering completely the surface of culture vessels. Almost all of the cultured cells, however, became chromophobic, at least at the end of the first week of cultivation, when gonadotrophin was detectable neither in the culture medium nor in the cells themselves. After the addition of the hypothalamic extract, gonadotrophin became detectable again, and basophilic or PAS-positive granules also reappeared within the cells, suggesting that the gonadotrophs were stimulated by the extract to produce gonadotrophin. In organ culture and short term incubation, the incorporation of [3H] leucine into the adenohypophysial cells in relation to the addition of hypothalamic extract was examined. It was obvious that the ability to incorporate [3H] leucine into the gonadotrophs in vitro was highly dependent upon the presence of the hypothalamic extract.

scholarly journals Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Hafiz Muhammad Umer Farooqi ◽  
Bohye Kang ◽  
Muhammad Asad Ullah Khalid ◽  
Abdul Rahim Chethikkattuveli Salih ◽  
Kinam Hyun ◽  
...  
Keyword(s):  
Cell Culture ◽  
Liver Fibrosis ◽  
Real Time ◽  
Hepatic Fibrosis ◽  
Transforming Growth Factor ◽  
Cell Monolayer ◽  
Embedded Sensors ◽  
Matrix Deposition ◽  
Tgf Β1

AbstractHepatic fibrosis is a foreshadowing of future adverse events like liver cirrhosis, liver failure, and cancer. Hepatic stellate cell activation is the main event of liver fibrosis, which results in excessive extracellular matrix deposition and hepatic parenchyma's disintegration. Several biochemical and molecular assays have been introduced for in vitro study of the hepatic fibrosis progression. However, they do not forecast real-time events happening to the in vitro models. Trans-epithelial electrical resistance (TEER) is used in cell culture science to measure cell monolayer barrier integrity. Herein, we explored TEER measurement's utility for monitoring fibrosis development in a dynamic cell culture microphysiological system. Immortal HepG2 cells and fibroblasts were co-cultured, and transforming growth factor β1 (TGF-β1) was used as a fibrosis stimulus to create a liver fibrosis-on-chip model. A glass chip-based embedded TEER and reactive oxygen species (ROS) sensors were employed to gauge the effect of TGF-β1 within the microphysiological system, which promotes a positive feedback response in fibrosis development. Furthermore, albumin, Urea, CYP450 measurements, and immunofluorescent microscopy were performed to correlate the following data with embedded sensors responses. We found that chip embedded electrochemical sensors could be used as a potential substitute for conventional end-point assays for studying fibrosis in microphysiological systems.

scholarly journals Building constructions of stone cladded roofs in contemporary architecture

2011 ◽  
Vol 9 (1) ◽  
pp. 35-56 ◽  
Author(s):  
Gergely Dobszay
Keyword(s):  
Work Experience ◽  
Building Envelope ◽  
Underlying Structure ◽  
Main Role ◽  
Visual Appearance ◽  
Expected Performance ◽  
Time Requirements ◽  
Under Construction ◽  
The Individual ◽  
Almost All

Stone roof coverings have existed for hundreds of years now, but their application in contemporary representative architecture is quite different than it was in former times, and accordingly the underlying constructional details are much more complicated. The main role of the roof in today's architecture is to emphasize the abstract building volume and its homogeneity. The design (visual appearance) has priority over constructional considerations, while at the same time requirements for thermal insulation and the watertight buildup of the building envelope are rising. This is a big challenge for the design of the constructional details, since traditional methods do not apply anymore: almost all of the water gets under this exterior cladding where a completely waterproof underlayer is needed. Therefore a new type of constructions is borne. In this article I try to present innovative constructional alternatives for stone cladded roofs and try to define guidelines for their planning. These alternatives were created and refined based on analysis of historical and contemporary examples, the examination of the specific conditions and requirements that affect cladded roofs, their expected performance over their life-span, the difficulties that arise under construction and my work experience. I created seven groups, according to the method for fixing the individual stone elements, since this is the detail that most affects the buildup of the underlying structure. A new approach is needed with these unconventional constructions, with a much more detailed inventory of every little feature of the building in question. But since these features vary violently with the architectural conception the exact constructional solutions are usually unique as well.

scholarly journals Galaxies hosting an active galactic nucleus: a view from the CALIFA survey

2020 ◽  
Vol 492 (3) ◽  
pp. 3073-3090 ◽  
Author(s):  
Eduardo A D Lacerda ◽  
Sebastián F Sánchez ◽  
R Cid Fernandes ◽  
Carlos López-Cobá ◽  
Carlos Espinosa-Ponce ◽  
...  
Keyword(s):  
Star Formation ◽  
Morphological Type ◽  
Molecular Gas ◽  
Main Role ◽  
Type I ◽  
Massive Galaxies ◽  
Star Forming ◽  
Integral Field Spectroscopy ◽  
Almost All ◽  
Formation Efficiency

ABSTRACT We study the presence of optically-selected active galactic nuclei (AGNs) within a sample of 867 galaxies extracted from the extended Calar-Alto Legacy Integral Field spectroscopy Area (eCALIFA) spanning all morphological classes. We identify 10 Type-I and 24 Type-II AGNs, amounting to ∼4 per cent of our sample, similar to the fraction reported by previous explorations in the same redshift range. We compare the integrated properties of the ionized and molecular gas, and stellar population of AGN hosts and their non-active counterparts, combining them with morphological information. The AGN hosts are found in transitory parts (i.e. green-valley) in almost all analysed properties which present bimodal distributions (i.e. a region where reside star-forming galaxies and another with quiescent/retired ones). Regarding morphology, we find AGN hosts among the most massive galaxies, with enhanced central stellar-mass surface density in comparison to the average population at each morphological type. Moreover, their distribution peaks at the Sab-Sb classes and none are found among very late-type galaxies (>Scd). Finally, we inspect how the AGN could act in their hosts regarding the quenching of star-formation. The main role of the AGN in the quenching process appears to be the removal (or heating) of molecular gas, rather than an additional suppression of the already observed decrease of the star-formation efficiency from late-to-early type galaxies.

CHARACTERIZATION OF DISORDERS OF PLATELET-VESSEL WALL INTERACTION IN AN AGGREGOMETER INCORPORATING BLOOD FLOW PAST AN ENDOTHELIAL CELL MONOLAYER

1987 ◽  
Author(s):  
E F Grabowski ◽  
K McKenny
Keyword(s):  
Endothelial Cell ◽  
Cell Monolayer ◽  
Fold Increase ◽  
Flow Chamber ◽  
Fluorescent Label ◽  
Endothelial Cell Monolayer ◽  
Flowing Blood ◽  
Normal States ◽  
Wall Interaction

Epi-fluorescence videomicroscopy permits real-time imaging of platelet (plt) adhesion-aggregation to a defined microinjury site of an endothelial cell monolayer (ECM) exposed to flowing blood. The fluorescent label is the TAB murine monoclonal antibody (courtesy of Dr. R.P. McEver) directed against human pit cp HB, together with a fluorescein-conjugated goat F(ab')2 against murine immunoglobulin. The combination assures specificity for pit membranes, yet leaves pit function intact. Bovine aortic ECM, grown on rectangular cover glasses, comprise one wall of a flow chamber mounted on a vertical microscope stage. A 6-0 sterile suture, drawn across the ECM in a direction transverse to flow, creates microinjuries of width 70 ± 15 (mean ± SD). Pit deposition is virtually absent upon intact and confluent regions of the ECM. On microinjury sites and at a shear rate of 270 sec-1, however, computer-enhanced images show pit adherence, aggregation, and embolization. Pretreatment of the ECM with 1.0 mMFC lysine acetyl salicylate, further, leads to a three-fold increase in aggregate length. ECM products inhibitable by aspirin, therefore, modulate adhesion-aggregation in disease and normal states under physiologic flow conditions. The Table shows that nercent coverage of the injury area, and mean aggregate length readily discriminate normal, post-aspirin, and von Willebrand's (vWD's) bloods. Aggregate length is reduced in vWD's blood to a greater degree (p<0.01) than by oral aspirin, while the latter is associated with a paradoxic increase (p<0.01) in single plt adhesion.

scholarly journals armadillo, bazooka, and stardust are critical for early stages in formation of the zonula adherens and maintenance of the polarized blastoderm epithelium in Drosophila.

1996 ◽  
Vol 134 (1) ◽  
pp. 149-163 ◽  
Author(s):  
H A Müller ◽  
E Wieschaus
Keyword(s):  
Plasma Membranes ◽  
Basolateral Membrane ◽  
Cell Sheet ◽  
Cell Monolayer ◽  
Test System ◽  
Sem Analysis ◽  
Drosophila Embryo ◽  
Early Stages ◽  
A Cell ◽  
The Stability

Cellularization of the Drosophila embryo results in the formation of a cell monolayer with many characteristics of a polarized epithelium. We have used antibodies specific to cellular junctions and nascent plasma membranes to study the formation of the zonula adherens (ZA) in relation to the establishment of basolateral membrane polarity. The same approach was then used as a test system to identify X-linked zygotically active genes required for ZA formation. We show that ZA formation begins during cellularization and that the basolateral membrane domain is established at mid-gastrulation. By creating deficiencies for defined regions of the X chromosome, we have identified genes that are required for the formation of the ZA and the generation of basolateral membrane polarity. We show that embryos mutant for both stardust (sdt) and bazooka (baz) fail to form a ZA. In addition to the failure to establish the ZA, the formation of the monolayered epithelium is disrupted after cellularization, resulting in formation of a multilayered cell sheet by mid-gastrulation. SEM analysis of mutant embryos revealed a conversion of cells exhibiting epithelial characteristics into cells exhibiting mesenchymal characteristics. To investigate how mutations that affect an integral component of the ZA itself influence ZA formation, we examined embryos with reduced maternal and zygotic supply of wild-type Arm protein. These embryos, like embryos mutant for both sdt and baz, exhibit an early disruption of ZA formation. These results suggest that early stages in the assembly of the ZA are critical for the stability of the polarized blastoderm epithelium.

Studies on pathogenic, immunogenic, and protective efficiency of fox rabies virus before and after adaptation to cell culture: application to vaccination against rabies

1983 ◽  
Vol 29 (1) ◽  
pp. 77-83 ◽  
Author(s):  
J. Blancou ◽  
M. F. A. Aubert ◽  
L. Andral
Keyword(s):  
Cell Culture ◽  
Rabies Virus ◽  
Oral Route ◽  
Submandibular Salivary Gland ◽  
Protective Efficiency ◽  
Safety Testing ◽  
Lethal Infection ◽  
Erroneous Diagnosis ◽  
Before And After ◽  
High Doses

Rabies virus from the submandibular salivary gland of a naturally infected fox was adapted to growth in BHK-21 cells. The pathogenicity of the original isolate and the cell culture adapted virus were compared by the intramuscular and oral routes in mice and foxes. Animals surviving exposure were tested for serum rabies antibodies (immunogenic efficiency) and for their ability to survive a second challenge with rabies virus (protective efficiency). In mice, ratios between lethal and protective doses of the two strains via the oral or muscular route were similar. In foxes, however, pathogenic efficiency was modified after cell culture adaptation, although good immunogenic and protective efficiency was maintained. Inoculation via the oral route resulted in lethal infection with the wild-type strain and survival (with some subsequent immunity) with the cell-adapted strain. Via the intramuscular route, foxes given high doses of cell culture adapted virus survived whereas several foxes given lower doses died and virus could not be reisolated. It is concluded that safety testing of modified strains should be done with different dilutions of virus in the target species. "Autosterilizing" infections may result in erroneous diagnosis.

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