scholarly journals Reduced Latency in the Metastatic Niche Contributes to the More Aggressive Phenotype of LM8 Compared to Dunn Osteosarcoma Cells

Sarcoma ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Matthias J. E. Arlt ◽  
Ingo J. Banke ◽  
Josefine Bertz ◽  
Ram Mohan Ram Kumar ◽  
Roman Muff ◽  
...  
Keyword(s):  
Time Course ◽  
Kidney Tissue ◽  
Experimental System ◽  
Osteosarcoma Cells ◽  
Primary Tumors ◽  
Liver And Kidney ◽  
Time Course Study ◽  
Cell Dormancy

Metastasis is the major cause of death of osteosarcoma patients and its diagnosis remains difficult. In preclinical studies, however, forced expression of reporter genes in osteosarcoma cells has remarkably improved the detection of micrometastases and, consequently, the quality of the studies. We recently showed that Dunn cells equipped with alacZreporter gene disseminated from subcutaneous primary tumors as frequently as their highly metastatic subline LM8, but only LM8 cells grew to macrometastases. In the present time-course study, tail-vein-injected Dunn and LM8 cells settled within 24 h at the same frequency in the lung, liver, and kidney of mice. Furthermore, Dunn cells also grew to macrometastases, but, compared to LM8, with a delay of two weeks in lung and one week in liver and kidney tissue, consistent with prolonged survival of the mice. Dunn- and LM8-cell-derived ovary and spine metastases occurred less frequently.In vitro, Dunn cells showed less invasiveness and stronger contact inhibition and intercellular adhesion than LM8 cells and several cancer- and dormancy-related genes were differentially expressed. In conclusion, Dunn cells, compared to LM8, have a similar capability but a longer latency to form macrometastases and provide an interesting new experimental system to study tumor cell dormancy.

Suitability of Glomus intraradices in vitro produced spores and root segment inoculum for the establishment of a mycorrhizosphere in an experimental microcosm

2001 ◽  
Vol 79 (8) ◽  
pp. 879-885
Author(s):  
M Filion ◽  
M St-Arnaud ◽  
C Guillon ◽  
C Hamel ◽  
S H Jabaji-Hare
Keyword(s):  
Microbial Ecology ◽  
Time Course ◽  
Glomus Intraradices ◽  
Mycorrhizal Fungus ◽  
Experimental System ◽  
Arbuscular Mycorrhizal ◽  
Root Segment ◽  
Soil Conditions ◽  
Time Course Study

Various experimental systems have been developed to study the mycorrhizosphere. In this study, a microcosm experimental system was constructed and optimized to simulate the environments of the mycorrhizosphere: the rhizosphere, the mycosphere, and the bulk soil, using beans (Phaseolus vulgaris L.) as host plants. We investigated, in a time-course study, the effect of axenically in vitro produced spore inoculum and root segment inoculum of the arbuscular mycorrhizal fungus, Glomus intraradices Schenck & Smith, on extraradical mycelium development, rapidity of mycorrhizal colonization, and plant growth under nonsterile soil conditions. Three concentrations of in vitro produced spores and three concentrations of root segment inoculum produced from open pot cultures were used. The two highest concentrations of spores used as inoculum resulted in faster and more abundant colonization than when root segments were used. A significant correlation was obtained between hyphal densities present in the rhizosphere and mycosphere compartments, and the amount of spore inoculum used. The densities of roots in the rhizosphere compartment and hyphae in the rhizosphere and mycosphere compartments were comparable with field-grown plants; thus, the system realistically mimics a natural mycorrhizosphere. The use of the microcosm described in this study, in combination with the in vitro produced spore inoculum of G. intraradices, represents an experimental approach well adapted for studying the microbial ecology of the mycorrhizosphere.Key words: AMF, microbial ecology, inoculum, mycorrhiza, mycorrhizosphere.

Effects of 2,4-D on Metabolism of 14C-Glucose in Plant Tissues

Weed Science ◽  
1971 ◽  
Vol 19 (3) ◽  
pp. 248-253
Author(s):  
I. Y. Mostafa ◽  
S. C. Fang
Keyword(s):  
Time Course ◽  
Pentose Phosphate ◽  
Insoluble Residue ◽  
Opposite Pattern ◽  
Corn Roots ◽  
Pea Roots ◽  
Time Course Study ◽  
Acid Pathway ◽  
Dichlorophenoxy Acetic Acid

A time course study on the in vitro effect of 10–4 M (2,4-dichlorophenoxy)acetic acid (2,4-D) on the metabolism and incorporation of specific 14C-labeled glucose into pea (Pisum sativum L., var. Alaska) and corn (Zea mays L., var. Golden Cross) tissues showed a preferential release of C-1 as CO2 which was affected by 2,4-D. The glucuronic acid pathway was stimulated greatly in pea roots and slightly in corn stems; it was inhibited in corn roots. The pentose phosphate pathway was affected in an opposite pattern. The incorporation of both labeled carbon atoms into alcohol-insoluble residue was also affected by 2,4-D. The degree of effect, however, varied from pea to corn and from roots to stems. Analysis of the alcohol-soluble fraction revealed different effects of 2,4-D on labeling of some of the amino acids, an increase in fructose content, and an accumulation of malic acid.

155 QUALITY OF PORCINE EMBRYO PRODUCED IN TWO EMBRYO CULTURE MEDIA AS ASSESSED BY TOTAL CELL NUMBER AND TIME COURSE OF BLASTOCYST HATCHING

2006 ◽  
Vol 18 (2) ◽  
pp. 185 ◽  
Author(s):  
Y. Agca ◽  
H. Men ◽  
S. F. Mullen ◽  
L. K. Riley ◽  
R. S. Prather ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Time Course ◽  
Culture Media ◽  
Cell Number ◽  
Total Cell ◽  
Total Cell Number ◽  
Blastocyst Hatching ◽  
Hatching Rates

The ability to produce porcine embryos of good quality will have a significant impact on a number of porcine assisted reproductive technologies, such as cloning, intracytoplasmic sperm injection, and embryo cryopreservation. However, porcine embryos resulting from current serum-free embryo culture systems differ significantly both structurally and functionally from those derived in vivo (Wang et al. 1999 Mol. Reprod. Dev. 53, 99-107). In this experiment, the quality of porcine embryos produced by North Carolina State University (NCSU)-23 medium (Petters and Wells 1993 J. Reprod. Fertil. Suppl. 1993, 48, 61-73) and porcine zygote medium (PZM)-1 (Yoshioka et al. 2002 Biol. Reprod. 66, 112-119) were compared by assessing the total cell number and the time course of in vitro blastocyst hatching. Porcine embryos were produced by in vitro maturation and fertilization using serum-free systems. After fertilization, presumptive zygotes were randomly allocated to either PZM-1 or NCSU-23 for subsequent development. On Day 4 of culture, the embryo culture media were supplemented with 10% fetal bovine serum (FBS). Day 6 blastocysts from each group were counted and the blastocysts were subsequently fixed in 4% formalin for counting the total cell number. The cell number in each embryo was determined by counting the nuclei after staining with bisbenzimide (Hoechst 33342). To assess the hatching ability of blastocysts, Day 6 blastocysts were cultured until Day 9 and hatched blastocysts were counted daily. Day 6 blastocyst rates (ratio of blastocysts to oocytes) and total cell number count were replicated three times. The time course of blastocyst hatching experiment was repeated four times. The data were analyzed using a chi-square test, Fisher's exact test, or Student's t-test. The blastocyst rate from culture in PZM-3 was 19.4 � 0.96% (mean � SEM), which was similar to that (16.7 � 3.2%) resulting from culture in NCSU-23 (P > 0.05). However, the total cell number in Day 6 blastocysts cultured in PZM-3 was significantly higher than for blastocysts cultured in NCSU-23 (57 � 3.1 vs. 46 � 1.7; P < 0.01). The total hatching rates (ratio of hatched blastocysts to total blastocysts) by Day 9 were similar between the two culture systems (50.1 � 9.1% vs. 50.7 � 4.1%; P > 0.05). However, on Day 6, 2.1% of blastocysts from PZM-3 culture hatched whereas no blastocysts from NCSU-23 culture hatched. The cumulative hatching rates from PZM-3 culture on Day 7 were significantly higher than those from NCSU-23 culture (15.1 � 3.8% vs. 2.6 � 1.1%; P < 0.01). In conclusion, these data suggest that blastocysts produced in PZM-3 medium have better quality than blastocysts produced in the NCSU-23 culture system as assessed by the total cell number and the time course of blastocyst hatching. This project was supported by a grant from the National Institutes of Health (U42 RR 018877).

Time course of the meiotic arrest in sheep cumulus–oocyte complexes treated with roscovitine

Zygote ◽  
2015 ◽  
Vol 24 (2) ◽  
pp. 310-318 ◽  
Author(s):  
Letícia Ferrari Crocomo ◽  
Wolff Camargo Marques Filho ◽  
Camila Louise Ackermann ◽  
Daniela Martins Paschoal ◽  
Midyan Daroz Guastali ◽  
...  
Keyword(s):  
Exposure Time ◽  
Cell Expansion ◽  
Time Course ◽  
Cumulus Cell ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Nuclear Maturation ◽  
Maturation Medium

SummaryTemporary meiosis arrest with cyclin-dependent kinases inhibitors has been proposed in order to improve the quality of in vitro matured oocytes. In sheep, however, this phenomenon has been rarely investigated. Therefore, the present study aimed to evaluate the effect of different incubation times with roscovitine on nuclear maturation and cumulus cell expansion of sheep cumulus–oocyte complexes (COCs). For this, COCs were cultured for 0, 6, 12 or 20 h in basic maturation medium (Control) containing 75 μM roscovitine (Rosco). After, they were in vitro matured (IVM) for 18 h in the presence of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). At the end of each treatment, cumulus cell expansion and nuclear maturation were assessed under a stereomicroscope and by Hoechst 33342 staining, respectively. In the Control and Rosco groups, the absence of cumulus cell expansion prevailed at 0, 6, 12 and 20 h. After IVM for 18 h, total cumulus cell expansion in the Rosco treatments was dependent on the exposure time to roscovitine. A significantly high percentage of oocytes treated with roscovitine for 6 h (87%), 12 h or 20 h (65%) were arrested at the germinal vesicle (GV) stage. In contrast, 23% GVBD, 54% metaphase I (MI) and 61% MII oocytes were observed in the Control groups at 6, 12 and 20 h, respectively. In all treatments, a significant percentage of oocytes reached MII after IVM for 18 h. Therefore, roscovitine reversibly arrested the meiosis of sheep oocytes during different culture times with the maximal efficiency of meiotic inhibition reached at 6 h. In addition, reversibility of its inhibitory action on cumulus cells was exposure-time dependent.

scholarly journals The regulation of lipogenesis in vivo in the lactating mammary gland of the rat during the starved-refed transition. Studies wtih acarbose, a glucosidase inhibitor

1987 ◽  
Vol 242 (1) ◽  
pp. 235-243 ◽  
Author(s):  
S W Mercer ◽  
D H Williamson
Keyword(s):  
Mammary Gland ◽  
Time Course ◽  
Chow Diet ◽  
Glucosidase Inhibitor ◽  
Lactating Mammary Gland ◽  
Phosphofructokinase Activity ◽  
Time Course Study ◽  
Fructose 1,6 Bisphosphate

Depression of carbohydrate digestion by oral administration of acarbose, a glucosidase inhibitor, led to a 75% inhibition of the re-activation of lipogenesis in vivo in the mammary gland of 18 h-starved lactating rats refed with 5 g of chow diet. Rates of [1-14C]glucose incorporation in vitro into lipid and CO2 in mammary-gland acini isolated from refed animals were elevated compared with acini from starved rats, but acarbose treatment completely prevented this stimulation. Gastric intubation of glucose led to a large stimulation of lipogenesis in the mammary gland of starved lactating rats, similar to that induced by refeeding with chow diet; this was dependent on the amount of glucose given and the time elapsed between glucose administration and injection of 3H2O for the measurement of lipogenesis. The switch-on of lipogenesis in the mammary gland of starved lactating rats, by refeeding or by intubation of glucose, was associated with a decrease in the ratio of [glucose 6-phosphate]/[fructose 1,6-bisphosphate] in the gland, indicative of an increase in phosphofructokinase activity. A time-course study revealed that the ratio decreased rapidly over the first 30 min of chow refeeding, after which a large surge in lipogenesis was seen. Acarbose, given 25 min after the onset of refeeding, led to a stepwise increase in the ratio, in parallel with the observed decrease in lipogenic activity. It is concluded that the control of lipogenesis in the mammary gland is closely linked to the availability of dietary carbohydrate. An important site of regulation of lipogenesis in the gland appears to be at the level of phosphofructokinase. A possible role of insulin in the regulation of phosphofructokinase activity, and the acute modulation of insulin-sensitivity in the gland during the starved-refed transition, are discussed.

AVP and dDAVP in rabbit cortical collecting tubule: a comparative time-course study

1990 ◽  
Vol 258 (1) ◽  
pp. R99-R103
Author(s):  
M. Leite ◽  
W. N. Suki
Keyword(s):  
Time Course ◽  
Cell Types ◽  
Receptor Activation ◽  
Osmotic Water ◽  
Significant Difference ◽  
Time Course Study ◽  
Time Required ◽  
Collecting Tubules ◽  
Different Cell Types

The V2-selective analogue of arginine vasopressin (AVP), dDAVP, has been used to distinguish between the effects of V1- and V2-receptor activation by AVP in different cell types of the kidney. Based on studies showing different effects of AVP and dDAVP on prostaglandin secretion, and also on cytosolic Ca2+, we designed a comparative time-course study of both agonists on rabbit microdissected cortical collecting tubules (CCT) microperfused in vitro at 38 degrees C. Plots of the effects of AVP (10 microU/ml or 2.2 x 10(-11) M and 100 microU/ml or 2.2 x 10(-10) M) and dDAVP (10 microU/ml or 0.8 x 10(-11) M) on osmotic water permeability (Pf) at comparable antidiuretic activities, revealed an increase of Pf that was maintained for as long as 170 min of hormone exposure. Also the magnitude of increase in Pf and the time required to achieve the more sustained phase of response were comparable, with no significant difference between the two agonists. These results clearly demonstrate a stable response of rabbit CCT to AVP and dDAVP at physiological temperature, and they reveal no evidence for a difference between the native hormone AVP and its V2 selective analogue on the net hydrosmotic response of the CCT.

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