scholarly journals Pattern and frequency of nocodazole induced meiotic nondisjunction in oocytes of mice carrying the ‘tobacco mouse’ metacentric Rb(16.17)7Bnr

1995 ◽  
Vol 66 (1) ◽  
pp. 35-43 ◽  
Author(s):  
C. A. Everett ◽  
J. B. Searle
Keyword(s):  
High Frequency ◽  
Specific Effect ◽  
Specific Pattern ◽  
Similar Response ◽  
Biphasic Response ◽  
Meiotic Nondisjunction ◽  
High Level ◽  
Dose Dependent Increase ◽  
Dose Dependent

SummaryOocytes from (C3H/HeH × 101/H)F1 and Rb(16.17)7Bnr homozygous females were exposed to a range of doses of nocodazole in vitro. The spindle poison caused a dose dependent increase in metaphase I (MI) arrest and hyperploidy. A concentration of 0·03 μg/ml was found to induce a maximum hyperploid frequency of 3·1% and 11·6% respectively without a high level of MI arrest. Between 0·03 and 0·05 μg/ml MI arrest increased substantially and reached a frequency of approximately 90%. In a further experiment oocytes from Rb7 homozygous, heterozygous and 3H1 females were exposed to 0·03 μg/ml nocodazole 4, 6 or 8 h after the onset of maturation. The phase at which the spindle was inhibited resulted in a specific pattern of nondisjunction which in turn was dependent on whether the female carried an Rb metacentric. 3H1 oocytes gave a normally distributed pattern of increase in aneuploid frequency (over the spontaneous value) centering around a 6 h application. This was thought to be due to the interaction of chromosomes with the microtubules of the spindle during attachment and/or alignment. In contrast both Rb homozygotes and heterozygotes gave the same biphasic response, with a high frequency of aneuploidy in the oocytes when nocodazole was applied 4 and 8 h after the onset of maturation. In Rb homozygotes we demonstrated that the Rb bivalent underwent nondisjunction more frequently than the average acrocentric, when nocodazole was administered early. It can be assumed that the Rb trivalent in Rb heterozygotes showed a similar response. This early Rb specific effect, in combination with a delayed-version of the acrocentric effect found in the 3H1 mice was thought to generate the biphasic pattern. We discuss the implications of (a) the different meiotic behaviours of metacentrics and acrocentrics and (b) the meiotic delay in Rb mice.

Stimulation of HCO3- transport in isolated proximal bullfrog duodenum by prostaglandins

1980 ◽  
Vol 239 (3) ◽  
pp. G198-G203 ◽  
Author(s):  
G. Flemstrom
Keyword(s):  
Electrical Potential ◽  
Potential Difference ◽  
Electrical Potential Difference ◽  
Morphological Examination ◽  
Minimal Concentration ◽  
Dependent Transport ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Stimulation Of

An in vitro preparation of proximal duodenum from the bullfrog transported alkali into the luminal solution (approximately 1 mueq x h-1 x cm-2) and generated a transepithelial electrical potential difference (5-10 mV, lumen negative). Transport was inhibited by 2,4-dinitrophenol (10(-5) M), CN- (5 X 10(-3) M), indomethacin (5 X 10(-5) M), and acetazolamide (5 X 10(-3) M) indicating that metabolism is required. Both alkali transport and the electrical potential difference showed a dose-dependent increase on administration of the prostaglandins E2, 16,16-dimethyl E2, and F2 alpha. The minimal concentration stimulating transport was lower with the E-type prostaglandins (10(-8) M than with F2 alpha (10(-6) M), and the former also produced greater maximal responses. In addition to metabolic-dependent transport of alkali, there was passive transmucosal migration of HCO3-, amounting to approximately 40% of basal (unstimulated) transport and sensitive to variation of the transmucosal hydrostatic pressure. Morphological examination showed that the preparation is devoid of Brunner glands. Stimulation of duodenal epithelial HCO3- transport by prostaglandins may contribute to their previously demonstrated ability to prevent duodenal ulceration.

Metabolic and Pharmacologic Interaction of Ethanol and Metronidazole in the Rat

1972 ◽  
Vol 50 (6) ◽  
pp. 476-484 ◽  
Author(s):  
H. Kalant ◽  
A. E. LeBlanc ◽  
M. Guttman ◽  
J. M. Khanna
Keyword(s):  
Incidental Finding ◽  
Repeated Administration ◽  
Volatile Material ◽  
Alcohol Dehydrogenase Activity ◽  
Liver Homogenates ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Concentration Curves ◽  
Central Depressant

Metronidazole, added in vitro, did not act either as an inhibitor or as a substrate for the alcohol dehydrogenase activity of rat liver homogenates. Concentration curves of ethanol and acetaldehyde in the blood after an oral dose of ethanol were not altered by pretreatment with metronidazole; in contrast, disulfiram caused marked elevation of acetaldehyde levels. When given once only, metronidazole (or possibly a metabolite of it) exerted a mild central depressant effect of its own and produced a dose-dependent increase in the intoxicant effect of ethanol. After repeated administration of metronidazole, synergism with ethanol was not seen. An incidental finding was the production of a volatile material during incubation of solutions containing NAD, which gives an acetone-like peak in gas-liquid chromatograms.

scholarly journals Effect of glucose on fermentation heat in sheep rumen fluid in vitro

1986 ◽  
Vol 56 (1) ◽  
pp. 305-311 ◽  
Author(s):  
A. Arieli
Keyword(s):  
Heat Production ◽  
High Glucose ◽  
Infinite Dilution ◽  
Rumen Fluid ◽  
Heat Production Rate ◽  
Sheep Rumen ◽  
Effect Of Glucose ◽  
Dose Dependent Increase ◽  
Dose Dependent

1. Heat production rate (H) of rumen fluid was measured in a direct calorimeter, Basal H of samples of 15 ml rumen fluid mixed with 45 ml buffer was 0.4 mW/ml rumen fluid.2. Addition of glucose (0.4–6.4 mg/sample) was followed by a dose-dependent increase in H. Maximal H was 1.1 rnW/ml and lasted up to 5 min, returning thereafter to the basal level.3. Expression of fermentation heat (Hf; kJ/mol substrate added) against glucose dose indicated an asymptotic dose response.4. Maximal Hf(at infinite dilution) agreed with stoichiometric calculations whereas minimal Hfsuggested a partial fermentation of the substrate at a high-glucose dose in the rumen environment.

scholarly journals Establishment and characterization of human B-cell lymphoma cell lines using B-cell growth factor

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1311-1318 ◽  
Author(s):  
RJ Ford ◽  
A Goodacre ◽  
I Ramirez ◽  
SR Mehta ◽  
F Cabanillas
Keyword(s):  
Growth Factor ◽  
Cell Growth ◽  
B Cell ◽  
Cell Lines ◽  
Lymphoma Cells ◽  
B Cell Growth Factor ◽  
Hodgkin's Lymphomas ◽  
Dose Dependent Increase ◽  
Dose Dependent

B-cell non-Hodgkin's lymphomas (NHL-B) have been difficult to establish in long-term cell culture using standard techniques. We report the establishment of five representative cell lines from high grade NHL-B using B-cell growth factor (BCGF). The five NHL-B cell lines display the morphologic, immunophenotypic, genotypic, and biologic characteristics of the lymphoma cells present in the original diagnostic specimen. The cell lines showed at least a sevenfold dose- dependent increase in proliferation in vitro over background in the presence of BCGF. Other putative B-cell growth-stimulating cytokines showed no significant proliferative activity or were inhibitory in some cases. NHL-B cell lines secreted growth factor(s) into culture supernatants that mediated at least a fivefold dose-dependent increase in cell proliferation in autochthonous lymphoma cells and a 10-fold or greater stimulation in growth factor-dependent normal B cell lines in vitro. The cell lines show monoclonal rearrangements of IgH genes and nonrandom chromosomal abnormalities characteristic of NHL-B, while the expression of Epstein-Barr virus associated antigen (EBNA-I) is present in two of the five cell lines. The studies show that lineage-specific growth factors may be used to establish neoplastic B cell lines in vitro, which are important experimental systems for cellular and molecular studies in the NHL-B.

Immunolocalization, binding, and biological activity of endothelin in rabbit uterus: effect of ovarian steroids

1991 ◽  
Vol 260 (2) ◽  
pp. E292-E305
Author(s):  
M. Maggi ◽  
G. B. Vannelli ◽  
A. Peri ◽  
M. L. Brandi ◽  
G. Fantoni ◽  
...  
Keyword(s):  
Endothelin Receptors ◽  
Binding Studies ◽  
Stromal Compartment ◽  
Ovarian Steroids ◽  
Specific Receptors ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Two Populations ◽  
Dissociation Constant Kd

Specific immunostaining for endothelin 1 (ET-1) was observed in the endometrium but not myometrium of rabbits. The staining was dramatically affected by subacute treatment with ovarian steroids: epithelial cells were predominantly positive in immature rabbits, whereas, in sex steroid-primed rabbits, ET-1 was mainly localized in the stromal compartment. Binding studies were performed in myometrium of estrogen-treated rabbits using labeled ET-1 and ET-3, the corresponding unlabeled peptides, and sarafotoxin b (SRTX). Mathematical modeling of experimental results indicates that two populations of sites are present in myometrium. One site (R1 = 1 pmol/mg protein) shows approximately the same affinity for ET-1, ET-3, and SRTX [dissociation constant (Kd) 100 pM], whereas the second site (R2 = 10 pmol/mg protein) selectively binds ET-1 (Kd 400 pM). According to binding studies, ET-1 was more potent than SRTX in stimulating uterine contraction "in vitro." The subacute administration of increasing concentrations of 17 beta-estradiol (0.2-200 micrograms/kg for 4 days), but not 17 beta-estradiol (200 micrograms/kg for 4 days) plus progesterone (5 mg/kg for 4 days), stimulates a dose-dependent increase in endothelin receptors in myometrium (half-maximal effective dose = 0.7 micrograms/kg for 4 days). However, estrogen treatment does not affect the concentration of endothelin receptors in myometrial cells in primary culture. Conversely, divalent ions like calcium and magnesium enhance the binding of ET-1 to both uterine membranes and cells. Our results indicate that in rabbit uterus endothelin is present in the endometrium, whereas specific receptors are located in myometrium.

355 DEVELOPMENTAL COMPETENCE OF IMMATURE BUBALINE CUMULUS - OOCYTE COMPLEXES VITRIFIED BY THE OPEN PULLED STRAW AND CONVENTIONAL STRAW METHODS

2007 ◽  
Vol 19 (1) ◽  
pp. 293
Author(s):  
A. Sharma ◽  
G. N. Purohit
Keyword(s):  
Morphological Changes ◽  
Developmental Competence ◽  
Control Group ◽  
Equal Concentration ◽  
Nuclear Maturation ◽  
Developmental Capacity ◽  
Phosphate Buffered Saline ◽  
Dose Dependent Increase ◽  
Dose Dependent

The in vitro maturation (IVM), fertilization (IVF), and morphological changes in buffalo cumulus–oocyte complexes (COCs) cryopreserved by ultrarapid freezing using conventional (CON) and open pulled staw (OPS) methods were tested. COCs were cryopreserved using a vitrification solution comprised of Dulbecco's phosphate-buffered saline+0.5 M sucrose+0.4% BSA and two concentrations (4.5 or 5.5 M) of each cryoprotectant ethylene glycol (EG) and dimethylsulfoxide (DMSO) by either the CON or the OPS method. Vitrified COCs were stored in LN for 7 days and then thawed; morphologically normal COCs were used for IVM (n = 1070) and IVF (n = 933) in 2 separate experiments to record morphological damage of COCs due to vitrification, nuclear maturation 24 h after culture (9 replicates), and fertilization 24 h after insemination (10 replicates). The COCs were matured in vitro in TCM-199 media with hormone supplements and fertilized using TALP-BSA as described previously (Purohit et al. 2005 Anim. Reprod. Sci. 87, 229–239). Freshly collected COCs were separately used for IVM (n = 110) and IVF (n = 130) and kept as controls. The arcsin transformed data of the proportions of oocytes matured or fertilized was compared by Duncan's new multiple range test. The highest proportion of morphologically normal oocytes was seen in 5.5 M EG with the CON method (94.5%) and the lowest was seen in 4.5 M DMSO with the OPS method (82.4%). At the end of experiment 1, it was apparent that IVM in all vitrification groups was significantly lower (P < 0.05) compared to the control group (66.4%). Among the various vitrification treatments, the highest IVM occurred in 5.5 M EG with the OPS method (39.2%) and the lowest in 4.5 DMSO with the CON method (19.3%). Comparison of both concentrations of EG and DMSO showed that the proportion of COCs attaining Metaphase-II (M-II) increased with increasing concentration of both of the cryoprotectants. However, at equal concentration of EG and DMSO, the proportion of COCs attaining M-II was significantly higher in the OPS method compared to the CON method. In experiment 2, a significantly higher (P < 0.05) IVF was seen for fresh COCs (45.4%) compared to vitrified COCs. Among the vitrification treatments, the highest fertilization was seen in 5.5 M EG with the OPS method (33.6%) and the lowest in 4.5 M DMSO with the CON method (15.17%). A dose-dependent increase in the proportion of oocytes fertilized was seen with increasing concentration of both EG and DMSO [CON: 4.5 M (15.2%), 5.5 M (25.6%); OPS: 4.5 M (21.3%), 5.5 M (27.5%)] in both CON and OPS methods. Comparison of the 2 cryoprotectants revealed that EG was better compared to DMSO.At equal concentrations of EG or DMSO, a significantly higher (P < 0.05) proportion of fertilized oocytes was seen in the OPS method compared to the CON method. It was concluded that vitrification results in some damage to oocytes, with decrease in their subsequent IVM and IVF. Developmental capacity of vitrified buffalo oocytes can be improved by using OPS instead of conventional straws.

Effect of Tumor Necrosis Factor-α on in vitro Prostaglandin Production in Buffalo Corpus Luteum

2021 ◽  
Author(s):  
M.K. Tripathi ◽  
S. Mondal ◽  
I.J. Reddy ◽  
A. Mor
Keyword(s):  
Mrna Expression ◽  
Corpus Luteum ◽  
Prostaglandin E ◽  
Prostaglandin F ◽  
Important Regulator ◽  
Factor Α ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
Necrosis Factor

Background: Corpus luteum plays key role in embryonic survival. Prostaglandins are the important regulator controlling the life span of corpus luteum. The present study investigated the effect of various doses of TNFα on in vitro PGF2α and PGE2 production and expression profiling of PGFS and PGES mRNA in buffalo Corpus Luteum (CL).Methods: Buffalo ovaries with mid-luteal phase CL were collected from the abattoir and CL were enucleated from surrounding tissues. Corpus luteum were finely chopped, rinsed with HBSS (Hanks Balanced Salt Solution) medium; supplemented with gentamycin and 0.1% BSA and incubated at 37°C for 1 hr in HBSS containing 0.1% collagenase. The cell suspension following filtration was washed by HBBS supplemented with gentamycin and 0.1% BSA (bovine serum albumin) and was treated with increasing doses of TNFα (0.1, 0.5 and 1.0 nM) and cultured at 38.5°C, 5% CO2 level for 24 hr. Result: There was dose dependent increase in concentrations of PGF2α and PGE2 with increasing doses of TNFα. The PGFS (prostaglandin F synthase) mRNA expression increased with increasing doses of TNFα. However, there was decrease in PGES (prostaglandin E synthase) mRNA expression at 0.1 nM and 0.5 nM TNFα but PGES mRNA expression increased at 1.0 nM TNFα as compared to control. It can be concluded that TNFα may alter PGES and PGFS mRNA expression and prostaglandin secretion in buffalo CL. 

Effect of five fungicides on growth and ochratoxin A production by two Aspergillus carbonarius and Aspergillus niger isolated from Moroccan grapes

2014 ◽  
Vol 4 (3) ◽  
pp. 118-126
Author(s):  
Souad Zouhair ◽  
Souad Qjidaa Qjidaa ◽  
Atar Selouane ◽  
Driss Bouya ◽  
Cony Decock ◽  
...  
Keyword(s):  
Growth Rate ◽  
Ochratoxin A ◽  
Fungal Growth ◽  
Inhibitory Effect ◽  
Aspergillus Carbonarius ◽  
Total Inhibition ◽  
Germination And Growth ◽  
Dose Dependent Increase ◽  
Dose Dependent

Five fungicides azoxystrobin (ortiva), benomyl (benlate), hexaconazole (hexa), pyrimethanil (scala) and thiabendazole (tectocal) were tested sepa-rately in vitro for their ability to inhibit the growth of two ochratoxigenic strains of A. niger and A. carbonarius previously isolated from grapes. All fungicides effectively reduced the growth rate of A. carbonarius and A. niger from 34 to 100% at the recommended dose (RD). Thiabendazole caused total inhibition of spore germination and growth of the two strains, regardless of the doses assayed. Benomyl completely inhibited growth of A. niger whereas for A. carbonarius, concentrations above 0.02xRD were required to prevent the growth. The inhibitory effect of hexaconazole, azoxystrobin and pyrime-thanil was dose-dependent. At sub-lethal concentrations of three fungicides, a dose-dependent increase in in ochratoxin A biosynthesis by two strains was observed. The use of fungicide should be checked for its ability to inhibit fungal growth as well as for their effect in terms of mycotoxins biosynthesis.

Carcinoembryonic Antigen (CEA) Regulates Tumor-Angiogenesis in Colon Carcinomas.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1897-1897
Author(s):  
Kira Braemswig ◽  
Marina Poettler ◽  
Wazlawa Kalinowska ◽  
Christoph Zielinski ◽  
Gerald W Prager
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Tumor Angiogenesis ◽  
Endothelial Cell Proliferation ◽  
Erk Phosphorylation ◽  
Dose Dependent Increase ◽  
Dose Dependent

Abstract Human carcinoembryonic antigen (CEA) is a cell surface adhesion molecule member of the Immunoglobulin Superfamily (IgSF). Aberrant upregulation and secretion of soluble CEA is a common feature found in a wide variety of human cancers such as colon, breast and lung. Previous in vitro and in vivo results have demonstrated that CEA can affect tumor cell behavior including the inhibition of cell differentiation and apoptosis. However, any functional effects on angiogenic endothelial cell behavior are so far unknown. In the present work we found that in endothelial cells exogenous CEA led to a time and dose dependent increase in ERK phosphorylation, which was inhibited by the specific MEK inhibitor U0126. Thereby, the observed CEA effect was comparable in time and intense with the canonical angiogenic growth factor VEGF. The CEA-induced ERK phosphorylation was not affected by the blockage of VEGFR-2 / flk-1 using a specific inhibiting peptide (CBO-P11), which indicates a VEGF-independent mechanism. Furthermore, co-stimulation of endothelial cells with VEGF and CEA shows synergistic effects on ERK phosphorylation. While in endothelial cells no endogenous expression of CEA is detected, its putative receptor, the CEA receptor (CEAR), is highly expressed as shown by immunohistochemical staining of paraffin-embedded colon carcinoma sections as well as in biochemical analyses. When an activating antibody against CEAR was used, CEA-induced ERK phosphorylation was mimicked, while downregulation of CEAR by siRNA diminished CEA-induced signal transduction, significantly. To test a biological relevance of our findings, we first measured endothelial cell proliferation: CEA led to a dose dependent increase in endothelial cell proliferation in vitro, which again revealed a synergistic effect with VEGF. Thereby, CEA-induced endothelial cell proliferation was again independent of VEGFR-2 / flk-1. A biological role of CEA in tumor-angiogenesis was reflected by an in vivo model using CEA Mimotope immunized BALB/c mice, which were transplanted with MethA/CEA overexpressing tumor cells. Immunohistological analyses of these tumors revealed a significantly reduced vascular density, which was accompanied with diminished tumor growth. Our data provide first evidence of CEA as a novel pro-angiogenic activator of endothelial cells, which results in an increase in endothelial cell proliferation, independent of VEGFR-2. Furthermore, by targeting CEA in an in vivo mouse model, tumor-angiogenesis was markley reduced, indicating a potential therapeutic target in cancer.

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