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Biomedical subjects

M Marx

Publications and source records attributed to M Marx.

At least 127 records · Page 7Linked to original sources
PubMed1990

Transformation-defective mutants with 5' deletions of the src gene are frequently generated during replication of Rous sarcoma virus in established quail fibroblasts.

Replication of Rous sarcoma virus (RSV) in avian fibroblasts leads to the generation of replication-competent variants that are defective for cell transformation (td virus). These td variants contain deletions affecting various portions of the v-src gene. We compared the rate of td virus production in Q3B cells, a quail cell line established by mutagen treatment, and in normal quail fibroblasts. Twenty-five days after infection with an RSV stock containing only transforming virions, Q3B cells harbor similar amounts of v-src-containing and v-src-deleted proviruses. However, these cells synthesize very low levels of p60v-src and generate large excess of td variants, as determined by biological assays. Unlike Q3B cells, normal quail fibroblasts infected with the same virus stock produce td variants only after multiple passages of undiluted virus on fresh cells. Restriction analysis showed that the td virus produced by Q3B cells is composed of two types of genomes: one lacking the entire v-src gene and the other carrying partial deletions of this gene predominantly located in the amino-terminal portion of the coding region of v-src. To study the mechanisms of these partial deletions, we molecularly cloned and sequenced the v-src genes of several td proviruses. We show that these mutants carry single or multiple v-src deletions of limited size, presumably generated by multiple mechanisms. Two deletions of 170 and 112 bp located in the 5' portion of v-src are frequently generated during RSV replication in Q3B cells and may represent preferential sites for v-src deletion in these cells.

Animals↗
PubMed1990

Additive effects of calcium antagonists on cyclosporin A-induced inhibition of T-cell proliferation.

The purpose of this study was to investigate possible additive effects of calcium antagonists on the cyclosporin A (CsA)-induced inhibition of cellular immunity. Human T-cells were isolated using standard methods and stimulated with phytohaemagglutinin (PHA, n = 8), the monoclonal antibody OKT3 (n = 6), or mixed lymphocyte reaction (MLR, n = 5). Verapamil, nifedipine, nimodipine or diltiazem were added (5 x 10(-7) - 5 x 10(-5) M) to the cultures, either alone, or in combination with CsA (62.5, 125, and 250 ng/ml). 3H-thymidine uptake was measured to estimate the proliferative responses and dose response curves were constructed for the Ca antagonists and their combinations with CsA. A 50% inhibition of T-cell proliferation in the different stimulation assays was achieved with 3.2 x 10(-5) - 5.3 x 10(-5) M verapamil, 2.5 x 10(-5) -4.3 x 10(-5) M nifedipine, 3.7 x 10(-6) - 5 x 10(-6) M nimodipine, and greater than 5 x 10(-5) M diltiazem. In combination with CsA a dose-dependent additive inhibitory effect of the Ca antagonists on T-cell proliferation was observed. This effect was less pronounced in the OKT3 assay, intermediate after PHA stimulation and most pronounced in MLR. Even in low concentrations, which correspond to therapeutic serum concentrations, Ca antagonists have an additive inhibitory effect in MLR. We conclude that Ca antagonists exert a dose-dependent inhibitory effect on T-cell proliferation. A combination of CsA with verapamil, nifedipine, nimodipine, or diltiazem is more effective than each drug given alone. This additive effect of Ca antagonists and CsA may possibly contribute to a better graft survival in clinical transplantation.

Adult↗
PubMed1990

Urinary secretory immunoglobulin A in acute renal allograft rejection.

Periodical determinations of the urinary secretory immunoglobulin A (S-IgA) excretion rate were performed in 12 cadaveric graft recipients. In five patients with primary functioning grafts the S-IgA excretion on the first postoperative day was 4.2 +/- 2.6 mg/g creatinine, decreasing to 1.8 +/- 1.2 mg/g creatinine (P less than 0.05) at the day of discharge. Acute tubular necrosis developed in the seven remaining patients. In this group the initial S-IgA excretion was 12.6 +/- 7.5 mg/g creatinine (P less than 0.05 compared to the former group), decreasing to 2.0 +/- 0.9 mg/g creatinine (P less than 0.05) at discharge. An acute rejection episode was observed in six patients. The S-IgA excretion increased from 3.0 +/- 1.5 mg/g creatinine 3-4 days before rejection to 6.4 +/- 3.1 mg/g creatinine (P less than 0.05) 1-2 days before rejection, and peaked at 14.0 +/- 8.6 mg/g creatinine (P less than 0.05) when the diagnosis of rejection was established and anti-rejection treatment was started. In three patients the initial steroid pulse therapy was not successful and S-IgA excretion further increased to 29.0 +/- 15.6 mg/g creatinine. After successful anti-rejection treatment, using steroids and OKT3, the S-IgA excretion decreased to 3.4 +/- 2.6 mg/g creatinine. In acute graft rejection, the elevated globulin synthesis by infiltrating plasma cells. In the early phase of rejection, dimeric IgA is the only immunoglobulin able to penetrate into the urine by transepithelial transport after binding to secretory component expressed on tubular epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Molecular and biological properties of c-mil transducing retroviruses generated during passage of Rous-associated virus type 1 in chicken neuroretina cells.

IC1, IC2, and IC3 are novel c-mil transducing retroviruses generated during serial passaging of Rous-associated virus type 1 (RAV-1) in chicken embryo neuroretina cells. They were isolated by their ability to induce proliferation of these nondividing cells. IC2 and IC3 were generated during early passages of RAV-1 in neuroretina cells, whereas IC1 was isolated after six consecutive passages of virus supernatants. We sequenced the transduced genes and the mil-RAV-1 junctions of the three viruses. The 5' RAV-1-mil junction of IC2 and IC3 was formed by a splicing process between the RAV-1 leader sequence and exon 8 of the c-mil gene. The 5' end of IC1 resulted from homologous recombination between gag and mil sequences. Reconstitution experiments showed that serial passaging of IC2 in neuroretina cells also led to the formation of a gag-mil-containing retrovirus. Therefore, constitution of a U5-leader-delta c-mil-delta RAV-1-U3 virus represents early steps in c-mil transduction by RAV-1. This virus further recombined with RAV-1 to generate a gag-mil-containing virus. The three IC viruses transduced the serine/threonine kinase domain of the cellular gene. Hence, amino-terminal truncation is sufficient to activate the mitogenic property of c-mil. Comparison of the transforming properties of IC2 and IC1 showed that the transduced mil gene, expressed as a unique protein independent of gag sequences, was weakly transforming in avian cells. Acquisition of gag sequences by IC1 not only increased the rate of virus replication but also enhanced the transforming capacity of the virus.

Animals↗
PubMed1990

Experience with long-term nifedipine therapy in paediatric cardiological patients.

Clinical experience with long-term nifedipine treatment in 23 patients aged between 1 1/12 and 14 8/12 years is reported. The cardiopulmonary diseases comprised primary pulmonary diseases with pulmonary hypertension (n = 4), congenital heart defects with intracardiac shunts and pulmonary hypertension which either were inoperable as a result of an Eisenmenger reaction (n = 7) or presented a high surgical risk (n = 5), or defects in which pulmonary hypertension did not regress despite corrective (n = 1) or palliative surgery (n = 3), and congenital defects without pulmonary hypertension (n = 3). Subjective improvement with an increase in physical performance was clearly observed in 15 cases. Echocardiography and cardiac catheter examinations showed no progression of the pulmonary arterial diseases, except in 1 patient with severe primary pulmonary hypertension and an 11-year observation period with nifedipine treatment during the last 4 years. No complications occurred during the 4 corrective operations. A patient aged 14 8/12 years with the Down syndrome and atrioventricular septal defect developed easily controllable heart failure during 7-day administration of nifedipine without additional cardiotherapy. 4 children initially suffered from flushed face and scalp, in one case with headache; 2 children reported fatigue. Long-term treatment with nifedipine should begin with strict 7-day supervision in hospital and possibly additional digitalization. Success of the treatment was determined by an improved quality of life in patients with primary pulmonary hypertension and inoperable defects, and by a reduced perioperative risk and postoperative regression of pulmonary hypertension in patients with operable defects.

Adolescent↗
PubMed1989

Healing at graft fixation site under functional conditions in posterior cruciate ligament reconstruction. A morphological study in sheep.

The incorporation course at the fixation sites of a free patellar tendon graft used in posterior cruciate ligament reconstruction was examined in eight adult sheep. After surgery the sheep were functionally treated without any protection of the operated legs. Two, 6, 12, 16, and 26 weeks after surgery the sheep were killed and the character of biological attachment to bone at fixation sites was evaluated using macroradiography, high-resolution radiography, and routine histology. After 6 weeks good osseous incorporation of the bone pegs of free patellar tendon grafts could be demonstrated at the desired position.

Animals↗
PubMed1989

Secretory immunoglobulin A in urinary tract infection, chronic glomerulonephritis, and renal transplantation.

An enzyme-linked immunoassay (ELISA) for the quantitation of secretory IgA (S-IgA) in the urine is described. The assay is sensitive (linear down to 0.02 microgram/ml), specific, and reproducible (intraassay variation 7.8%, interassay variation 18.9%). The assay was used to determine the urinary excretion rates of S-IgA in patients with bladder catheter (n = 12), cystotomy and urinary tract infection (n = 13), urosepsis (n = 5), chronic glomerulonephritis, including IgA-nephritis (n = 31), in renal allograft recipients (n = 30), and in healthy controls (n = 46). An elevated S-IgA urinary excretion (P less than 0.01) was found in all patient groups studied. The highest values were observed in urosepsis. No significant differences between S-IgA levels in IgA-nephritis and other forms of chronic glomerulonephritis were detected. The elevated S-IgA excretion in renal allograft recipients indicates that immunosuppressive therapy does not influence the local S-IgA production in the urinary tract. The susceptibility of these patients to urinary tract infections is therefore not due to a S-IgA deficiency.

Diuresis↗
PubMed1989

Short-duration, high-dose urokinase infusion for recanalization of occluded saphenous aortocoronary bypass grafts.

Thrombolytic recanalization of arterial bypass grafts has been pursued aggressively in the peripheral circulation but not in the coronary circulation. In an attempt to apply peripheral transcatheter thrombolytic techniques to the coronary circulation, nine patients with 10 occluded saphenous aortocoronary bypass grafts underwent recanalization procedures using a short-duration, high-dose urokinase infusion. Urokinase was infused at the occluded graft orifice at a rate of 600 units/min. The average infusion time was 1 hr, 26 min. The average urokinase dose was 435,000 units. Graft recanalization was achieved in eight (80%) of 10 grafts, although only six (60%) of 10 grafts were widely patent at the end of the procedure. All successfully recanalized grafts required balloon angioplasty of underlying stenoses. No complications, specifically myocardial infarction or cerebrovascular accident, were encountered. We have shown that occluded aortocoronary bypass grafts can be recanalized successfully by using a short-duration, high-dose urokinase infusion. It appears that, with attention given to angiographic techniques that minimize clot manipulation, recanalization can be accomplished safely in a majority of cases.

Adult↗
PubMed1988

Stable propagation of the active transcriptional state of an immunoglobulin mu gene requires continuous enhancer function.

Enhancer sequences control the expression of the immunoglobulin mu heavy chain gene. In this study we examined whether the active transcriptional state of the mu gene can be propagated from mother to daughter cells after deletion of its enhancer. A mu gene that contains the enhancer flanked by D and J recombination signals was stably introduced into pre-B cells, resulting in mu gene transcription. Subsequent deletion of the enhancer from the transfected gene by D-to-J joining reproducibly resulted in switching off of mu expression. Moreover, transcriptional inactivation occurred although the mu gene was found to be hypomethylated prior to deletion of the enhancer. We conclude that the enhancer does not confer upon the mu gene "memory" of its active transcriptional state. Thus, the enhancer seems to be required both for establishment and for maintenance of mu gene expression.

Base Sequence↗
PubMed1988

[Plasma thiocyanate determination: a simple method for quantifying the smoking habits of pregnant patients--a preliminary study].

Plasma thiocyanate (SCN) was measured by means of a simple colorimetric method [8] in 328 persons (175 males, 153 females, aged 5-68 years) as part of an ongoing prospective trial on the influence of SCN defined maternal smoking on fetal development. The interassay coefficient of variation for synthetic K thiocyanate solutions was 3.1% (n = 25%); the intrassay coefficient of variation was 1.2% (n = 15). Mean SCN levels were significantly lower in 226 nonsmokers (SCN 41.4 +/- 18.5 mumol/l) than 102 smokers (SCN 81.4 +/- 28.7 mumol/l; p less than 0.01). Of all the variables tested, a direct influence on SCN was found only in the case of diabetes mellitus and haemolytic blood samples (p less than 0.01), whereas age, sex, chronic diseases and nutrition had no effect on SCN. Mean maternal post partum SCN levels (n = 48) were significantly lower in non-smokers (30.2 +/- 11.2) than in smokers (59.3 +/- 23.1). There was a linear relationship between maternal venous plasma and fetal umbilical cord plasma SCN values (r = 0.88; p less than 0.01). Pregnant women had lower SCN levels than non-pregnant women. We conclude that SCN is an easy means of measuring and a reproducible parameter for quantifying smoking behaviour in pregnancy and in the general population.

Adolescent↗
PubMed1988

A novel oncogene related to c-mil is transduced in chicken neuroretina cells induced to proliferate by infection with an avian lymphomatosis virus.

Non-dividing neuroretina cells from chicken embryos are induced to proliferate after a long latency, following infection with Rous associated virus type 1, an avian retrovirus which does not carry a transforming gene. We have isolated from these proliferating cells an acutely mitogenic retrovirus, designated IC10, which contains a novel oncogene. Nucleotide sequencing showed that the IC10 virus has transduced 1101 nucleotides of cellular origin inserted between the gag and env genes of RAV-1. This oncogene, designated v-Rmil, is 70.1% homologous to v-mil. v-Rmil encodes a protein of 40,976 daltons sharing 83.8% homology with the catalytic domain of the v-mil protein. Divergence with the v-mil gene product is observed at the NH2- and COOH-terminal portions of the v-Rmil protein. Restriction analysis of normal chicken DNA indicated that v-Rmil is derived from a cellular gene distinct from c-mil. The c-Rmil gene is transcribed through a major mRNA, greater than 10 kb in length, that is detected at much higher levels in neuroretinas, as compared to other embryonic tissues.

Amino Acid Sequence↗
PubMed1988

The effects of various indwelling ureteral catheter materials on the normal canine ureter.

In an attempt to objectively evaluate the biocompatibility of materials commonly used for ureteral stenting, stent-catheters made of four different materials were placed randomly in 31 ureters of 19 mongrel dogs. Animals underwent urography at four weeks and were sacrificed at six weeks. Mild hydronephrosis was noted in eight instances, essentially unrelated to specific material. Silicone, C-flex and polyurethane stents caused a similar, mild degree of ureteral edema, but ureters stented with Silitek demonstrated fairly marked edema. Epithelial ulceration and erosion, often severe, occurred with all polyurethane stents, and rarely with the three other materials. All materials differed statistically from controls, and C-flex caused less reaction overall than polyurethane, indicating differences in biocompatibility of the various materials which may be relevant to their use in patients undergoing long-term ureteral stenting. In practical terms, these suggest that certain materials, notably silicone and C-flex, are more suitable for ureteral stenting than others.

Animals↗
PubMed1988

Activation and transduction of c-mil sequences in chicken neuroretina cells induced to proliferate by infection with avian lymphomatosis virus.

We report that nondividing neuroretina cells from chicken embryos can be induced to proliferate following infection with Rous-associated virus type 1 (RAV-1), an avian lymphomatosis retrovirus lacking transforming genes. Multiplication of RAV-1-infected neuroretina cells is observed after a long latency period and takes place initially in a small number of cells. We also show that serial virus passaging onto fresh neuroretina cultures leads to the generation of novel mitogenic viruses containing the mil oncogene. DNA analysis indicated that RAV-1 is the only provirus detected in cells infected at virus passage 1, whereas neuroretina cells infected at subsequent virus passages harbor mil-containing proviruses. Three viruses, designated IC1, IC2, and IC3, were molecularly cloned. Restriction mapping indicated that in each virus, truncated c-mil sequences were inserted within different portions of the RAV-1 genome. In addition, IC1 and IC2 viruses have transduced novel sequences that belong to the 3' noncoding portion of the c-mil locus. All three viruses induce neuroretina cell multiplication and direct the synthesis of mil-specific proteins. Proliferation of neuroretina cells infected at passage 1 of RAV-1 was not associated with any detectable rearrangement of c-mil, when a v-mil probe was used. However, these cells expressed high levels of an aberrant 2.8-kilobase mRNA hybridizing to mil but not to a long terminal repeat probe. Therefore, transcriptional activation of a portion of c-mil could represent the initial events induced by RAV-1 infection and lead to retroviral transduction of activated c-mil sequences.

Animals↗
PubMed1987

N-terminal deletion in the src gene of Rous sarcoma virus results in synthesis of a 45,000-Mr protein with mitogenic activity.

Expression of the v-src gene of Rous sarcoma virus in avian embryo neuroretina cells results in transformation and sustained proliferation of these normally resting cells. Transformed neuroretina cells are also tumorigenic upon inoculation into immunodeficient hosts. We have previously described conditional mutants of Rous sarcoma virus encoding p60v-src proteins which induce proliferation of neuroretina cells in the absence of transformation and tumorigenicity. These results suggest that p60v-src is composed of functionally distinct domains which may interact with multiple cellular targets. In this study, we describe a spontaneous variant of Rous sarcoma virus, subgroup E, which carries a deletion of 278 base pairs in the 5' portion of the v-src gene but which has retained the ability to induce proliferation of quail neuroretina cells. The deleted v-src gene encodes a 45,000-molecular-weight phosphoprotein which contains both phosphoserine and phosphotyrosine, is myristylated, and possesses tyrosine kinase activity indistinguishable from that of wild-type p60v-src. Molecular cloning and sequence analysis of the mutant v-src gene have shown that this deletion extends from amino acid 33 to 126 of the wild-type p60v-src. Therefore, this portion of the v-src protein is dispensable for the mitogenic activity of Rous sarcoma virus in neuroretina cells.

Animals↗