In vitro and in vivo association and subcellular distribution of toxic and experimentally modified endotoxin

1982 ◽  
Vol 28 (7) ◽  
pp. 822-829
Author(s):  
Hassan Tavakoli ◽  
Robert J. Moon
Keyword(s):  
Tissue Culture ◽  
Nuclear Transfer ◽  
Experimental System ◽  
Reticuloendothelial System ◽  
Organ Distribution ◽  
Culture Cells ◽  
Cell Association ◽  
Htc Cells

Cell association and organ distribution of toxic and experimentally modified endotoxin were compared in whole animals and hepatoma tissue culture (HTC) cells. For both toxic and poly-L-α-ornithine mixed endotoxin in vivo, most of the endotoxin becomes associated with the reticuloendothelial system (RES) rich organs. Organ distribution does not change from 1 to 5 h. Significantly less detoxified endotoxin becomes associated with RES-rich organs.Association and nuclear transfer of toxic endotoxin in HTC cells are gradual and time-dependent processes. Plasma treatment increased association of endotoxin with HTC cells. Poly-L-α-ornithine (4 μg/mL) also significantly increases HTC cell association of endotoxin, and nuclear transfer of endotoxin was similar in principle to the toxic material. Association of detoxified endotoxin with HTC cells is significantly higher than toxic endotoxin and increases with time. In contrast with toxic and poly-L-α-ornithine mixed endotoxin, nuclear association of alkaline-treated detoxified endotoxin did not increase significantly during 5 h of incubation.Cumulatively, these observations indicate that while tissue culture cells could provide a more controllable experimental system by which to study the fate and pathogenic mechanism of endotoxin at the cellular and subcellular level, HTC cells under the conditions employed herein do not yield binding data which compare favorably with in vivo results. Caution must be exercised when extrapolating in vitro data to the actual in vivo action of endotoxin.

scholarly journals ELECTRON MICROSCOPY OF PLASMA-CELL TUMORS OF THE MOUSE

1961 ◽  
Vol 9 (2) ◽  
pp. 369-381 ◽  
Author(s):  
D. F. Parsons ◽  
M. A. Bender ◽  
E. B. Darden ◽  
Guthrie T. Pratt ◽  
D. L. Lindsley
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Complex Structure ◽  
Granular Body ◽  
Virus Particles ◽  
Culture Cells ◽  
Large Numbers ◽  
Tumor Agent

The X5563 tumor has been grown in tissue culture. Cells similar to those of the original tumor migrated from the explant and attached to the glass walls of the culture vessels. Electron microscopy showed that large numbers of particles, similar in morphology to virus particles, were associated with these cells after 7 days of culture. The two principal types of particles found in the tumor in vivo appear to be present in vitro. Many more of these particles, however, were larger and showed a more complex structure. Whereas the particles were mainly localized inside endoplasmic reticulum or the Golgi zone in the tumors in vivo, in the tissue culture the majority of the particles were associated with the plasma membrane and were found outside of the cells. The relation of the particles to the granular body is discussed as well as a possible relation to the mammary tumor agent.

In Vitro and In Vivo Models of Spinal Muscular Atrophy

2017 ◽  
Author(s):  
W. David Arnold ◽  
Arthur H. M. Burghes
Keyword(s):  
Tissue Culture ◽  
Spinal Muscular Atrophy ◽  
Muscular Atrophy ◽  
In Vivo Models ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Smn Protein

Spinal muscular Atrophy (SMA) is caused by reduced levels of the SMN protein. In humans this is caused by loss of SMN1 and retention of SMN2. The challenge in modelling SMA, in either tissue culture cells or animals, is first to obtain the desired SMN levels equivalent to what is observed in SMA. Various models of SMA in tissue culture cells, invertebrates, and mammals have been created have been developed. The targets of SMN reduction that are most relevant for the pathogenesis of SMA and how the phenotype of SMA can be modified independent of SMN levels are two important questions that remain unanswered. Here the current in vitro and in vivo models of SMA are summarized.

Accumulation of Macromolecular Particles in the Reticuloendothelial System (RES) Mediated by Platelet Aggregation and Disaggregation

1969 ◽  
Vol 22 (03) ◽  
pp. 496-507 ◽  
Author(s):  
W.G van Aken ◽  
J Vreeken
Keyword(s):  
Platelet Aggregation ◽  
Degradation Products ◽  
Reticuloendothelial System ◽  
Caproic Acid ◽  
Carbon Particles ◽  
Carbon Clearance ◽  
Aggregation And Disaggregation ◽  
Platelet Aggregates

SummaryCarbon particles cause platelet aggregation in vitro and in vivo. Prior studies established that substances which modify thrombocyte aggregation also influence the rate at which carbon is cleared from the blood.This study was performed in order to elucidate the mechanism by which the carbon-platelet aggregates specifically accumulate in the RES.Activation of fibrinolysis by urokinase or streptokinase reduced the carbon clearance rate, probably due to generated fibrinogen degradation products (FDP). Isolated FDP decreased the carbon clearance and caused disaggregation of platelets and particles in vitro. Inhibition of fibrinolysis by epsilon-amino-caproic acid (EACA), initially accelerated the disappearance of carbon and caused particle accumulation outside the RES, predominantly in the lungs. It is supposed that platelet aggregation and locally activated fibrinolysis act together in the clearance of particles. In the normal situation the RES with its well known low fibrinolytic activity, becomes the receptor of the particles.

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

In vitro and in vivo pathologic effects of Vibrio parahaemolyticus on human epithelial cells

1984 ◽  
Vol 30 (3) ◽  
pp. 381-388 ◽  
Author(s):  
B. R. Merrell ◽  
R. I. Walker ◽  
S. W. Joseph
Keyword(s):  
Epithelial Cells ◽  
Epithelial Tissue ◽  
Ileal Mucosa ◽  
Cytotoxic Factor ◽  
Culture Cells ◽  
Vibrio Parahemolyticus ◽  
Buccal Epithelial Cells ◽  
Culture Supernate

The initial interaction and adherence of Vibrio parahemolyticus to epithelial tissue culture cells, human buccal epithelial cells, and the ileal mucosa of mice were studied. Using scanning electron microscopy, adherent bacteria were observed only on degenerating human embryonic intestinal, HeLa, and buccal cells; healthy normal cells were devoid of bacteria. Sheared V. parahaemolyticus, i.e., lacking flagella, did not adhere to either normal or degenerating tissue cells. Neither ultraviolet-inactivated organisms nor cell-free culture supernate affected the epithelial cells. Similar findings were observed on the mucosa of the ileum in mice inoculated with V. parahaemolyticus. It appears that V. parahaemolyticus possesses a cytotoxic factor which alters epithelial cells. This factor appears to be closely associated with viable organisms and may be a functional element in the adherence process of flagellated V. parahaemolyticus to mammalian epithelial cells.

In Vivo Interactions with (Tissue Culture Pretreated) Dermal Sheep Collagen

1991 ◽  
Vol 252 ◽  
Author(s):  
P. B. van Wachem ◽  
P. B. van Wachem ◽  
L. H. H. Olde Damink ◽  
P. J. Dijkstra ◽  
J. Feijen ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Cell Death ◽  
Marked Reduction ◽  
Giant Cells ◽  
In Vitro Cytotoxicity ◽  
Cell Infiltration ◽  
Cytotoxic Effects ◽  
Lipid Formation

ABSTRACTPretreatment in tissue culture (TC) was previously found to markedly reduce the in vitro cytotoxicity of two types of crosslinked dermal sheep collagens (DSC's). This in vivo study confirms our in vitro results, in that TC-pretreatment of crosslinked DSC's resulted in the marked reduction or elimination of cytotoxic effects, such as increased cell infiltration, a deviant neutrophil-morphology, lipid formation and cell death. TC-pretreatment affected the crosslinked state of both DSC's in a different way, which could be deduced from the differences in gelatin-formation and presence of giant cells from macrophage- or fibroblast-origin. The results are explained in view of the differences in crosslinking.

scholarly journals 64COMPARISON OF DEVELOPMENTAL POTENTIAL OF IN VIVO AND IN VITRO RECIPIENT OOCYTES AFTER NUCLEAR TRANSFER IN GOAT

2004 ◽  
Vol 16 (2) ◽  
pp. 154
Author(s):  
H.S. Park ◽  
M.Y. Lee ◽  
S.P. Hong ◽  
J.I. Jin ◽  
J.K. Park ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Electric Pulse ◽  
Serum Starvation ◽  
Cleavage Rate ◽  
Developmental Potential ◽  
In Vitro Development ◽  
Significant Difference ◽  
Vitro Development

Recent techniques in somatic cell nuclear transfer (SCNT) have been widely used for animal research. In addition, SCNT techniques may allow for the rescue of endangered species. Despite efforts for wildlife preservation, however, some threatened or endangered wild animal species will likely become extinct. As a preliminary experiment of a series in wildlife research, we tried to identify an improved method for the production of more transferable NT embryos in goats. Mature donor animals of Korean native goats (20–25kg) were synchronized with a CIDR (type G; InterAg, New Zealand) vaginal implant for 10 days followed by a total of 8 twice daily injections of 70mg of FSH (Folltropine, London, Ontario, Canada) and 400IU of hCG (Chorulon, Intervet, Moxmeer, The Netherlands). Oocytes were then collected surgically by retograde oviduct flush or direct aspiration from ovarian follicles in vivo at 29–34h after hCG. Oocytes collected from follicles were matured in TCM-199 containing 10% FBS and hormones. Prepared ear skin cells from the goat were cultured in TCM-199 containing 10% FBS at 39°C, 5% CO2 in air, and confluent monolayers were obtained. Oocytes were enucleated and donor cells from serum starvation (0.5%) culture were fused through a single electric pulse (DC 2.36kvcm−1, 17μs), and then activated by a single electric pulse (AC 5vmm−1, 5s+DC 1.56kvcm−1, 30μs) or chemical treatment (5μgmL−1 ionomycin 5min−1, 1.9mM 6-DMAP/4h). Reconstructed oocytes were cultured in M16 medium with 10% goat serum (GS) for 6–7 days. Data were analyzed by chi-square test. In in vitro development, significantly (P<0.05) more oocytes were cleaved (24/30, 80.0%) and developed (7/24, 29.2%) to morula or blastocyst stage, respectively, in NT oocytes activated by Iono + DMAP compared to electric stimulated oocytes (2/21, 40.0%; 0/2, 0%). There was a significant difference in in vitro development of NT embryos by the method of oocyte collection. Cleavage rate was higher (P<0.05) in NT embryos from in vivo oocytes (23/28, 82.1%) than in in vitro matured oocytes (19/35, 54.3%), and further development to morula or blastocyst was also significantly (P<0.05%) higher in NT embryos from in vivo oocytes (7/23, 30.4%) than in NT embryos from in vitro matured oocytes (0/19, 0%). When we compared NT embryos to parthenotes, developmental rate was not significantly different between NT embryos and parthenotes. These results strongly suggest that the in vivo oocytes will have superior developmental potential to oocytes matured in vitro. Table 1 Effect of different oocyte source on in vitro development following caprine SCNT

scholarly journals Effect of China berry crude extracts on broad mites in vivo and in vitro

2008 ◽  
Vol 2 (1) ◽  
pp. 12-18
Author(s):  
Assma Gatta ◽  
Luaay K. Al – ani ◽  
Nabeel Al - ani
Keyword(s):  
Biological Control ◽  
Tissue Culture ◽  
Melia Azedarach ◽  
Concentration Increase ◽  
Water Extracts ◽  
Crude Extracts ◽  
Number Increase ◽  
Ethanol Extracts

Tissue culture were established from leaf and stem of china berry (Melia azedarach ) tree . Using MS media the best regulator to form callus were 6mg/l BAP, all other concentrations did not give callus . The crude extracts from leaves and callus established from leaves were extracted with water and ethanol with different concentrations. In ethanol extracts the least concentration 0.0001 half of the treated parasites were killed in 24 hours while the number increase as the concentration increase . However in callus the ethanol extracts were much higher about 8.5 were killed in the above concentration . In water extracts the least concentration 0.0001 killed half of the treated parasites in 24 hours .This number was increased 8 or 9 in 48 and 72 hours respectively . These results give us preliminary idea about the biological control of this dangerous parasite.

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