ATP-related specific heterotrophic activity in petroleum contaminated and uncontaminated groundwaters

1989 ◽  
Vol 35 (8) ◽  
pp. 814-818 ◽  
Author(s):  
Bjørn K. Jensen
Keyword(s):  
Conversion Factor ◽  
Thymidine Incorporation ◽  
Specific Activity ◽  
Tritiated Thymidine ◽  
Atp Content ◽  
Heterotrophic Activity ◽  
Cell Counts ◽  
Plate Counts ◽  
Direct Counting ◽  
Activity Indices

Bacterial numbers, ATP content, heterotrophic potentials, and thymidine incorporation were determined for petroleum contaminated and uncontaminated groundwaters. ATP measurements and cell counts gave variations of a factor of 7 in values among the sites. The acridine orange direct counting yielded counts 2 to 3 orders of magnitude higher than the plate counts and counts 1 to 2 orders of magnitude higher than counts given by the ATP measurement using a conversion factor of 10−6 ng ATP/cell. Mean Vmax values for 14C-labled glucose, acetate, and naphthalene were 0.59, 0.17, and 1.14 μg ∙ L−1 ∙ h−1, respectively. Naphthalene was only used as substrate in the contaminated samples. Acetate and naphthalene showed fairly good correlations to ATP contents, 0.94 and 0.95, respectively, as well as with tritiated thymidine incorporation rates, r = 0.94. Vmax specific activity indices (SAI) based on ATP content showed very little variation for the selected groundwaters compared with SAI values reported earlier for groundwaters.Key words: groundwater, heterotrophic potential, tritiated thymidine incorporation, ATP.

Effect of inhibin on rat testicular desoxyribonucleic acid (DNA) synthesis in vivo and in vitro

1981 ◽  
Vol 98 (2) ◽  
pp. 312-320 ◽  
Author(s):  
P. Franchimont ◽  
F. Croze ◽  
A. Demoulin ◽  
R. Bologne ◽  
J. Hustin
Keyword(s):  
Dna Synthesis ◽  
Thymidine Incorporation ◽  
Specific Activity ◽  
Tritiated Thymidine ◽  
Biological Properties ◽  
Rete Testis ◽  
Thymidine Uptake ◽  
Adult Rats

Abstract. When injected in vivo 3 h before sacrifice or when incubated in vitro with testicular fragments for 3 h, tritiated thymidine, a reliable index of DNA synthesis and of mitotic activity, was incorporated into the DNA of differentiated spermatogonia, as shown by autohistoradiography. The maximum DNA specific activity was obtained in pubertal rats aged 42 days, weight 150 g. Two preparations of inhibin extracted from ram rete testis fluid (RTF) of different molecular weight (> 10 000 for RTF1 and < 5000 for RTF3) but which possess the same biological properties were investigated for their effect on thymidine uptake in vivo and in vitro. In vivo both preparations specifically inhibited tritiated thymidine incorporation into testicular DNA of pubertal animals (42 days). No change in thymidine uptake into hepatic DNA was observed. Tritiated thymidine incorporation into testicular DNA was lower in normal adult rats and in hypophysectomized pubertal animals. RTF1 and RTF3 did not affect thymidine incorporation in either case. The reasons for this lack of effect are discussed. In vitro, both preparations induced a dose-dependent decrease in DNA synthesis in testis fragments from rats aged 42 and 49 days. The preparations lost their in vivo and in vitro inhibitory effects when denatured by heating and trypsin digestion. The inhibin preparations probably reduced testicular DNA synthesis and spermatogonial multiplication by reducing FSH secretion in vivo but also had a direct effect on the germ cells as shown by the in vitro experiments. These in vivo and in vitro actions of inhibin preparations are similar to those of the testicular chalones. The relationship which might exist between inhibin and the chalones is discussed.

A STUDY OF BLASTOCYSTS DURING DELAY AND SUBSEQUENT IMPLANTATION IN LACTATING MICE

1968 ◽  
Vol 42 (3) ◽  
pp. 453-463 ◽  
Author(s):  
ANNE McLAREN
Keyword(s):  
Dna Synthesis ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Uterine Horn ◽  
Uterine Epithelium ◽  
Serial Sections ◽  
Uterine Lumen ◽  
Fresh State

SUMMARY Blastocysts were studied on the 5th and 8th day of pregnancy in lactating mice, in the fresh state, flushed from the uterus, in squash preparations and in serial sections. At the earlier period some mitosis was observed. Tritiated thymidine incorporation studies gave some evidence of DNA synthesis on the 5th and 6th days of pregnancy. By the 8th day the blastocysts were longer, contained more cells, and mitosis had ceased. They were located at the anti-mesometrial end of the uterine lumen, closely apposed to the uterine epithelium, and with their long axes parallel to the long axis of the uterine horn. Implantation could be induced, either by the removal of the litter, or by the injection of an appropriate dose of oestrogen on the 5th or 7th (but not the 4th) day of pregnancy. Both treatments were followed by the appearance of W-bodies in the neighbourhood of the blastocysts, the disappearance of the shed zonae, and the appearance of Pontamine Blue reactivity, oedema of the uterine stroma and formation of the primary decidual zone, in that order.

Interleukin 6 stimulates growth of vascular smooth muscle cells in a PDGF-dependent manner

1991 ◽  
Vol 260 (5) ◽  
pp. H1713-H1717 ◽  
Author(s):  
U. Ikeda ◽  
M. Ikeda ◽  
T. Oohara ◽  
A. Oguchi ◽  
T. Kamitani ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Interleukin 6 ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Muscle Cells ◽  
Thymidine Uptake ◽  
Dependent Manner

We have investigated the effect of interleukin 6 (IL-6) on the growth of vascular smooth muscle cells (VSMC) isolated from rat aortas. Murine recombinant IL-6 significantly increased the number of VSMC and stimulated tritiated thymidine incorporation into VSMC in a dose-dependent manner. The IL-6-induced thymidine incorporation into VSMC was totally inhibited by the Ca2+ channel blocker verapamil; however, IL-6 showed no effects on the intracellular Ca2+ level ([Ca2+]i) in VSMC. Antibody against platelet-derived growth factor (PDGF) also totally inhibited the IL-6-induced thymidine uptake. PDGF caused a significant increase in the [Ca2+]i, which was totally inhibited by verapamil. IL-6 mRNA was not detected in unstimulated “quiescent” VSMC, but its expression was stimulated by exposure of VSMC to 10% fetal bovine serum. Immunohistochemical study using anti-PDGF antibody showed that IL-6 stimulated PDGF production in VSMC. These results support the premise that IL-6 is released by VSMC in an autocrine manner and promotes the growth of VSMC via induction of endogenous PDGF production.

Timing of nucleolar DNA replication in Amoeba proteus

1976 ◽  
Vol 22 (3) ◽  
pp. 521-530
Author(s):  
I. Minassian ◽  
L.G. Bell
Keyword(s):  
Electron Microscope ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Central Region ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Amoeba Proteus ◽  
Electron Microscope Autoradiography ◽  
Microscope Autoradiography ◽  
G2 Phase

Light- and electron-microscope autoradiography have been used to follow the incorporation of [3H]thymidine at different stages during the interphase of synchronously growing populations of Amoeba proteus. Two main patterns were found for tritiated thymidine incorporation, i.e. DNA synthesis. The major incorporation was in the central region of the nucleus, but a lesser degree of incorporation occurred in the nucleolar region. The bulk of this nucleolar DNA was found to be late replicating, i.e. it replicated during the G2 phase.

VIP inhibits basal and histamine-stimulated proliferation of human airway smooth muscle cells

1995 ◽  
Vol 268 (6) ◽  
pp. L1047-L1051 ◽  
Author(s):  
K. Maruno ◽  
A. Absood ◽  
S. I. Said
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Thymidine Incorporation ◽  
Inhibitory Effect ◽  
Airway Smooth Muscle Cells ◽  
Dependent Manner ◽  
Cyclic Monophosphate ◽  
Cell Counts ◽  
Dependent Protein ◽  
Camp Levels

Airway smooth muscle (ASM) cell proliferation contributes to increased airway resistance in bronchial asthma. We have examined the modulation of ASM proliferation by vasoactive intestinal peptide (VIP), a cotransmitter of airway relaxation. Human ASM cells were grown in culture as a monolayer. VIP (1.0 nM-1.0 microM) inhibited proliferation in a dose-dependent manner by up to 82% on day 2, but the related peptide glucagon had no effect. Histamine (100 nM-100 microM) increased cell counts by 66%, but in the presence of VIP, cell counts and [3H]thymidine incorporation were reduced by up to 55%. Adenosine 3',5'-cyclic monophosphate (cAMP)-promoting agents, including 3-isobutyl-1-methylxanthine, forskolin, and 8-bromo-adenosine 3',5'-cyclic monophosphate, alone and especially combined with VIP, reduced cell counts and [3H]thymidine incorporation, in correlation with cAMP levels. KT-5720 (1.0 nM-1.0 microM), a selective inhibitor of cAMP-dependent protein kinase A (PKA), abolished the inhibitory effect of VIP. The results show that VIP selectively and potently inhibits human ASM cell growth and multiplication, and nullifies the mitogenic effect of histamine, by a PKA-mediated mechanism. A deficiency of VIP may lead to ASM hyperplasia due to unopposed stimulation by endogenous mitogens.

scholarly journals Changes in adipose tissue of the rat due to early undernutrition followed by rehabilitation

1980 ◽  
Vol 43 (1) ◽  
pp. 33-43 ◽  
Author(s):  
Janet Kirtland ◽  
Patricia M. Harris
Keyword(s):  
Adipose Tissue ◽  
Stromal Cell ◽  
Specific Activity ◽  
Tritiated Thymidine ◽  
Cell Replication ◽  
Fat Cell ◽  
Finite Period ◽  
Cell Fractions ◽  
Specific Radio ◽  
Deep Body

1. Well-nourished rats were injected with tritiated thymidine at 15, 22, 28 or 84 d of age. At 1, 6, 11 and 16 d after injection animals from each group were killed, samples of adipose tissue were removed from two subcutanecus sites (abdominal and scapular) and separated, using collagenase (EC 3.4.24.3), into ‘fat cell’ and ‘stromal cell’ fractions. The specific (radio)activity of DNA isolated from each fraction was measured. The specific activity of DNA isolated from two ‘deep body’ sites (perirenal and epididymal) was measured only in the animals injected at 84 d of age.2. Animals undernourished from birth up to 84 d of age were injected with tritiated thymidine at 22, 28 or 84 d of age. Animals were killed 1 and 11 d after injection, adipose tissue removed, and the specific activity of DNA measured. Other undernourished animals were rehabilitated from 84 to 107 d and injected at 91 d of age with tritiated thymidine. The animals were killed 1, 6, 11 and 16 d after injection, adipose tissue was removed from the subcutaneous and deep body sites and the specific activity of DNA determined as before.3. In well-nourished animals fat cell replication had largely ceased by 12 weeks of age in the subcutaneous depots. There were differences between the various sites of adipose tissue regarding the period of hyperplastic growth, its timing or rate of replication or both.4. In undernourished animals replication was slow in the subcutaneous depots compared with well-nourished animals of the same age. Rehabilitation from undernutrition stimulated replication which resulted in higher rates in all four depots examined compared with those in well-nourished animals.5. The findings are discussed in relation to the concept of a finite period of hyperplasia for adipose tissue.

scholarly journals Acidic isoferritins and E-type prostaglandins in sources of colony stimulatory factors mask detection of cycling granulocyte-macrophage progenitor cells

Blood ◽  
1982 ◽  
Vol 60 (4) ◽  
pp. 1042-1045 ◽  
Author(s):  
HE Broxmeyer
Keyword(s):  
Progenitor Cells ◽  
Conditioned Medium ◽  
Bone Marrow Cells ◽  
Specific Activity ◽  
Tritiated Thymidine ◽  
High Specific Activity ◽  
Prostaglandin E ◽  
Blood Leukocytes ◽  
Gm Csf ◽  
Feeder Layers

Abstract The effect of granulocyte-macrophage colony stimulatory factors (GM- CSF), acidic isoferritins, and E-type prostaglandins on the detection of the cycle status of human granulocyte-macrophage progenitor cells (CFU-GM) was investigated. Bone marrow cells were pulse-treated with control medium or high specific activity tritiated thymidine [3HTdr) and subsequently plated over feeder layers containing mononuclear blood leukocytes prepared in the absence or presence of anti-acidic isoferritins and/or indomethacin, or plated in the presence of medium conditioned by placental cell or GCT-conditioned media free of acidic isoferritins and prostaglandin-E. The presence of anti-acidic isoferritins and/or indomethacin in the blood leukocyte feeder layers increased the detectable stimulatory capacity of these cells and permitted detection of a larger proportion of marrow CFU-GM in cycle than in control cultures. The cycle status was not influenced by GM-CSF in conditioned medium regardless of the dilution of conditioned medium used to stimulate colony formation. This suggests that GM-CSF, supplied to the cells after treatment with 3HTdr, does not itself influence the detection of CFU-GM in cycle, but using sources of GM-CSF that contain acidic isoferritins or prostaglandin-E will underestimate the actual number of CFU-GM in S-phase.

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