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R Davies

Publications and source records attributed to R Davies.

At least 199 records · Page 11Linked to original sources

Sensitivity of the cell cycle to TGF beta 1 does not correlate with transformation of a rat liver epithelial cell line.

The effects of TGF beta 1 on cell cycle events in a rat liver derived epithelial cell line (BL9) and in two in vitro transformants of this line were studied by flow cytometry. Using either ethidium bromide staining or the incorporation of bromodeoxyuridine to evaluate DNA synthesis it was shown that TGF beta 1 prevented the entry of G0/G1 phase BL9 cells into S phase. TGF beta 1 did not exert its inhibitory effect(s) on DNA synthesis by the modulation of early events in the cell cycle. The tumorigenic transformed BL9 cell lines gave contrasting responses to the effects of TGF beta 1. DNA synthesis in a BL9 cell line derived by transfection with an active N-ras oncogene was unaffected by TFG beta 1 and thus appeared refractory to its growth controlling effects. On the other hand cells from a BL9 cell line derived by in vitro transformation with activated aflatoxin B1 retained their sensitivity to the effects of TGF beta 1. Thus the loss of the inhibitory effect of TGF beta 1 on DNA synthesis is not obligatory for the malignant transformation of rat liver epithelial cells.

Animals↗

A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis.

The mutation causing cystic fibrosis (CF) has been localized to the DNA sequence of 700 kb bounded by the loci identified by the markers pMP6d-9 (D7S399) and pJ3.11 (D7S8). A 560-kb fragment obtained after SacII digestion of DNA from a cell line containing this region of human chromosome 7 in a mouse background was separated using pulse-field gel electrophoresis and isolated from the gel. The DNA was digested with BamHI prior to cloning into lambda EMBL3. Approximately 0.1% of the resulting clones contained human repetitive sequences, and 24 such recombinants were studied. Of these, 23 are on chromosome 7; 8 clones were duplicated, and of the 15 different recombinants, 7 are between MET and INT1L1, and a further 7 are between INT1L1 and pMP6d-9, leaving a single marker, pG2, which is between pMP6d-9 and pJ3.11. pG2 recognizes an RFLP with XbaI. A cosmid walk from pG2 has generated a further marker, H80, which recognizes an RFLP with PstI. This new locus (D7S411) divides the remaining region between the CF flanking markers, thereby making it more accessible to fine pulse-field mapping and allowing the precise localization of further clones to this region. Although it is not possible to position the CF locus unequivocally with respect to D7S411, both polymorphic markers at this locus exhibit low but significant linkage disequilibrium with CF, placing the emphasis for the search for the gene on the D7S399 to D7S411 interval of 250 kb.

Blotting, Southern↗

Thyroid function in choriocarcinoma: demonstration of a thyroid stimulating activity in serum using FRTL-5 and human thyroid cells.

Hyperthyroidism is a well recognized complication of gestational trophoblastic tumours (GTT) and may be due to high circulating concentrations of human chorionic gonadotrophin (hCG) or its variants. We have studied 24 clinically euthyroid women with GTT. Eight were biochemically hyperthyroid with low or undetectable serum thyrotrophin (TSH) and had a mean serum hCG of 361.2 x 10(3) IU/l compared to 76.2 x 10(3) IU/l in the other patients (P less than 0.01). Purified hCG stimulated iodide uptake into FRTL-5 cells with 25 x 10(3) IU/l being equivalent in potency to 1 mU/l of thyrotrophin (TSH). Sixteen out of the 24 sera (67%) stimulated iodide uptake when applied to the cells at a 1:10 dilution. Sera from all eight hyperthyroid patients contained thyroid stimulating activity. The mean hCG concentration in the 16 stimulatory sera was 238.2 x 10(3) IU/l compared to 37.1 x 10(3) IU/l in the other eight sera (P less than 0.01). Six men with hCG-secreting testicular tumours were biochemically euthyroid although three of their sera stimulated iodide uptake into FRTL-5 cells. In human thyroid cells the mean cAMP production over 4 h with sera from five healthy controls was 54.2 +/- 1.81 pmol/mg cell protein compared to 67.0 +/- 3.8 pmol/mg protein with sera from five choriocarcinoma patients (P less than 0.02). Serum from patients with gestational trophoblastic tumours contains a thyroid stimulating activity which may be hCG and whose presence correlates with hyperthyroidism.

Adolescent↗

Effect of the DNA replication velocity on the response of Salmonella typhimurium to mild heating.

Changes in the DNA replication velocity of Salmonella typhimurium following mild heat stress (52 degrees C) were studied independently of the major physiological parameter of growth rate, using thymine-requiring mutant strains derived from Salm. typhimurium LT2. The isolated mutant strains BM1 or BM2, grown either as batch or chemostat cultures, showed a greater sensitivity to 52 degrees C heat stress when grown on a minimal medium containing near-limiting concentrations of thymine, compared with growth in the presence of excess thymine. Radiolabelling experiments provided evidence for alterations in the velocity of DNA replication upon growth on different thymine concentrations, independent of the growth rate. Thus, replicating DNA was implicated as the major site of injury after mild moist heat stress.

Culture Media↗

Survival of dehydrated cells of Salmonella typhimurium LT2 at high temperatures.

Cells of Salmonella typhimurium LT2 were dehydrated on hydrophobic membranes (Millipore FGLP2500) placed in a controlled atmosphere chamber held at 57% equilibrium relative humidity (ERH) and 37 degrees C. Dehydration for 48 h under the above conditions increased the heat resistance of Salm. typhimurium LT2 when measured as the surviving fraction after a heat challenge of 135 degrees C for 30 min. Results also showed that little or no death occurred during heat challenges of 1 h at temperatures of up to 100 degrees C. The survival of Salm. typhimurium LT2 was measured as the ability to form colonies on solid media tryptone soy broth plus 1.2% agar (TSBA) after 24 h at 37 degrees C. Incorporation of sodium pyruvate, at a concentration of (TSBA) after 24 h at 37 degrees C. Incorporation of sodium pyruvate, at a concentration of 0.2% into the recovery medium, did not enhance the recovery of heated Salm. typhimurium LT2. Dehydrated cells of S. typhimurium LT2 showed a triphasic death curve. Increasing the period of dehydration from 48 h to 34 d, reduced initial numbers due to die off but did not alter the shape of the subsequent survival curve.

Colony Count, Microbial↗

HLA-DR RFLP distributions in two groups of aboriginal Australians.

HLA-DR and -DQ typing by restriction fragment length polymorphism (RFLP) in Aboriginal Australians from the Kimberleys and from Coen shows a restricted number of HLA-DR types in these populations. The polymorphism is essentially limited to DR2, DR4, DRw14 and DRw8. The most common DR.DQ RFLP haplotype in Aborigines shows a novel arrangement of DR and DQ alleles that has important implications for histocompatibility matching if the RFLP patterns reflect functional variation in HLA class II molecules.

Australia↗

Thyroid-stimulatory effects of human chorionic gonadotrophin in early pregnancy. In vivo and in vitro studies.

Human chorionic gonadotrophin (hCG) shares structural similarity with pituitary thyrotrophin (TSH) and may act as a thyroid stimulator. We have studied serum hCG levels, thyroid function tests and the ability of serum to stimulate cultured thyroid cells in 40 subjects between 6 and 12 weeks of pregnancy. Serum free tri-iodothyronine was increased and serum TSH reduced in pregnancy samples (both p less than 0.05). hCG was detectable in all pregnancy sera with a mean level of 105.6 X 10(3) U/l. Serum from 24 of the 40 (60%) patients stimulated iodide uptake into cultured FRTL-5 thyroid cells. The potency of sera in stimulating cells correlated with the hCG level (r = 0.710, p less than 0.01). The stimulatory activity in some, but not all, sera could be specifically neutralized with antiserum to hCG. Partially purified hCG stimulated iodide uptake and growth of thyroid cells at concentrations of 50 X 10(3) U/l and above. In these experiments, 25 X 10(3) U/l of hCG produced equivalent stimulation to 1 mU/l of TSH. In 8 patients tested before and after termination of pregnancy, the thyroid-cell-stimulatory activity of serum declined rapidly in parallel with serum hCG. hCG may stimulate the thyroid gland at concentrations which prevail in normal pregnancy. Its potential as a physiological regulator of the thyroid gland is not widely appreciated and requires further study.

Adolescent↗

Transfection of cytochrome P450 cDNAs into mammalian cells used in mutation and transformation assays.

The present work demonstrates that cDNAs coding for cytochrome P450 enzymes can be transfected into mammalian cells and expressed. In the present studies, two different cell systems were used for transfection: 10T1/2 cells which can be used to study initiation and promotion (Diamond, 1984) and AHH-1 cells which can be used to study mutation and clastogenesis (Crespi and Thilly, 1984, Crespi and Penman, 1989). Thus, a diversity of endpoints can be studied in cells which have increased metabolic capability. By increasing the metabolic capability of the target cell, the effects of nongenotoxic as well as genotoxic chemicals, can be examined in the appropriate in vitro systems. For example, the 10T1/2 cells can be treated with a nontransforming dose of an initiator followed by continuous treatment with a second chemical that requires cytochrome P450 specific metabolism to manifest its promoting activity. By this approach, greater insight into the role of chemical metabolism in the promotion process (and presumably other nongenotoxic effects) can be obtained. Additionally, the role of specific cytochrome P450s in the metabolism of different classes of carcinogens/drugs can be elucidated. A major advantage of having the metabolizing enzymes actually present in the target cell is that effects of chemicals can be studied in long-term, low-dose exposure protocols which will eliminate the acute toxic effects which are associated with many current protocols. Thus, more realistic environmental exposure conditions can be achieved by using these in vitro systems containing endogenous metabolism systems.

Animals↗

Gd-DOTA. Pharmacokinetics and tolerability after intravenous injection into healthy volunteers.

The pharmacokinetics of Gd-DOTA meglumine in humans were evaluated in six healthy male volunteers. The agent was injected intravenously at 0.1 mmol/kg over approximately 2 minutes. Its behavior was found to be similar to that of urographic and angiographic iodinated contrast media with a plasma elimination half-life of 91 +/- 14 minutes (mean +/- standard deviation [SD]), a small distribution volume of 171.0 +/- 19.7 mL/kg and rapid urinary excretion. The results suggest rapid passive extravascular diffusion of gadolinium (Gd)-DOTA in the interstitial space without intracellular penetration, followed by a rapid urinary excretion via glomerular filtration. Furthermore, the results are consistent with animal data that showed that the compound does not cross the normal blood brain barrier. Its plasma pharmacokinetics appeared to be similar to those reported for Gd-DTPA. No relevant biological effects were seen with Gd-DOTA, especially in regard to serum iron and bilirubin levels.

Adult↗

Allergy.

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Adult↗

Cutting Körners.

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Appointments and Schedules↗

Persisting nystagmus following vestibular nerve section for Menière's disease.

Unilateral vestibular nerve section (VNS) creates a state of acute dysequilibrium which resolves by a process of central compensation. This disturbance resolves quickly and central compensation is complete generally within a month with resolution of symptoms and signs. The course of central compensation following VNS will be similar to that seen after labyrinthectomy because the detachment of hemi-labyrinthine input that is achieved by both will be identical. Six patients are presented who have undergone VNS at least 2.7 years ago (Average 3.5 years); all of them have persisting spontaneous peripheral type horizontal jerk nystagmus, present with optic fixation in five. This is obvious clinically and was confirmed in each case by agreement of three independent observers and has been recorded by electronystagmography (ENG). They are free from marked vestibular symptoms. The explanations of mechanisms involved in central compensation are discussed with respect to this previously unrecorded clinical observation.

Adult↗

Localization of a 23,000 MW antigen of Cryptosporidium by immunoelectron microscopy.

Rabbit antiserum was raised against a 23,000 molecular weight (MW) antigen prepared from Cryptosporidium oocysts by electro-elution from polyacrylamide gels. The antiserum was tested for specificity by immunoblotting against solubilized oocyst preparations. Several antigens including the 23,000 MW antigen were recognized suggesting that it shared common epitopes with higher MW proteins. The antiserum was then used in conjunction with a protein A-colloidal gold conjugate to locate antigenic sites within exogenous and endogenous developmental stages of Cryptosporidium. The pellicles of both sporozoites and merozoites exhibited specific labelling, particularly around their anterior ends. No specific labelling was observed for any other membrane determinants or organelles in these or other life cycle stages.

Adult↗

Oral and dermal pharmacokinetics of triclopyr in human volunteers.

Blood levels and urinary excretion of triclopyr, the active ingredient in Garlon herbicides, were followed in six volunteers given single oral doses of 0.1 and 0.5 mg/kg body weight. Five of these volunteers later received dermal applications of Garlon 4 herbicide formulation equivalent to 3.7 mg triclopyr/kg body weight applied to the forearm. Following oral administration blood levels peaked at 2-3 h and declined to undetectable levels within 48 h; more than 80% of the dose was found as unchanged triclopyr in the urine. A two-compartment pharmacokinetic model was used to describe the time-course of triclopyr clearance; half-lives for the rapid initial and slower terminal phases were 1.3 h and 5.1 h respectively, and were independent of dose. Due to the slow half-life for dermal absorption (t1/2 = 16.8 h) the rapid initial elimination phase was obscured and the pharmacokinetics could be simplified by a one-compartment model. An average of 1.37% of the applied dose was recovered in the urine; when corrected for recovery after oral administration this was equivalent to an absorption of 1.65%. Triclopyr is slowly absorbed through skin and is rapidly eliminated. It has very low potential to accumulate in man or to be absorbed through the skin in acutely toxic amounts.

Administration, Oral↗