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Biomedical subjects

R Snyder

Publications and source records attributed to R Snyder.

At least 109 records · Page 6Linked to original sources
PubMed1986

High molecular weight DNA from fixed cytogenetic preparations.

Cell pellets that have been stored in routine clinical cytogenetic fixative were studied for the presence of intact DNA. A method for the isolation of high molecular weight DNA from fixed cytogenetic preparations of human leukocytes, bone marrow, and cell hybrid cultures is presented. DNA preparations from fixed pellets were cleaved with restriction enzymes, transferred to nitrocellulose filters after agarose gel electrophoresis, and hybridized to radiolabeled probes to demonstrate that fixed cell pellets could yield DNA of sufficient quality for Southern blot hybridization analysis. This protocol may be useful for molecular analysis of DNA from fixed cell pellets of patients who are unavailable for additional sampling.

Bone Marrow↗
PubMed1985

Angiotensin-converting enzyme activity. A potential marker of tissue hypothyroidism in critical illness.

We measured serum angiotensin-converting enzyme (ACE) activity radiometrically as a possible indicator of reduced thyroid function in 57 euthyroid controls, 27 patients in a noncardiac intensive care unit (13 with medical and 14 with surgical disorders), and 29 patients having coronary artery bypass grafting. In the last group, blood was obtained preoperatively and one day and one month after surgery (group 1; n = 18) or preoperatively and six hours and one day after surgery (group 2; n = 11). Patients in group 1 had significant reductions in levels of serum thyroxine (T4), triiodothyronine (T3), and thyrotropin response to protirelin one day postoperatively. The ACE activity fell significantly. Patients in group 2 had low levels of T4, T3, thyrotropin, and ACE six hours postoperatively. All these levels remained low the next day, and free T4 and free T3 levels were also reduced; the reverse T3 level became elevated. Changes in ACE significantly paralleled changes in T3. The 27 patients without coronary artery bypass grafting also had significant reductions in serum T4, T3, and ACE levels. Dilution studies and dialysis of serum with low ACE activity failed to demonstrate an inhibitor to explain the reduced enzyme function.

Adult↗
PubMed1985

Inhibition of RNA synthesis by benzene metabolites and their covalent binding to DNA in rabbit bone marrow mitochondria in vitro.

Mitoplasts from rabbit bone marrow cells in vitro incubated with benzene metabolites showed a concentration-dependent inhibition of mitochondrial RNA synthesis. The 50% molar inhibitory concentration for each metabolite was determined to be p-benzoquinone, 2 X 10(-6); phenol, 2.5 X 10(-5); hydroquinone, 5 X 10(-5); catechol, 2 X 10(-3); benzene, 1.6 X 10(-2). Activated metabolites of radiolabeled benzene covalently bound to mitochondrial DNA in vitro. Labeled deoxynucleosides, enzymatically produced from DNA isolated from mitoplasts prelabeled in DNA with [3H]dGTP and exposed to [14C]benzene in vitro, were chromatographed on a Sephadex LH-20 column to obtain deoxynucleoside-adducts. The elution profiles indicated that mtDNA contained seven deoxyguanosine-adducts. A similar experiment in which the mtDNA was prelabeled with [3H]dATP indicated that two deoxyadenosine-adducts of mtDNA were formed from benzene in vitro. Tentative identification of several deoxyguanosine-adducts indicates that p-benzoquinone, hydroquinone, phenol, and 1,2,4-benzenetriol produced from benzene form adducts with guanine.

Animals↗
PubMed1985

A compound arterial prosthesis: the importance of the sterilization procedure on the healing and stability of albuminated polyester grafts.

In response to the demand for a vascular prosthesis which achieves reliable haemostasis without preclotting, a new compound albumin/polyester prosthesis has been developed. In order to optimize the sterilization procedures for this device, two series of implantations in the thoracic aorta of dogs were undertaken to compare the effects of ethylene oxide (EtO) and gamma-radiation. Preclotted polyester prostheses were implanted in two additional control series. Pathological analysis of the explanted grafts indicated that gamma-radiation is to be preferred over EtO because it results in faster rates of healing. While the albumin coating delayed the thrombotic response and fibrinolytic activity, the extent of healing of the radiation sterilized graft was equivalent to that achieved by preclotted polyester prostheses in the medium and long term. Measurements of the strength and dimensional changes of the graft demonstrated that, in addition to reducing the risks of acute thrombosis and postoperative haemorrhage, the albumin coating improves the dimensional stability of the knitted structure.

Albumins↗
PubMed1985

Inhibition of RNA synthesis and interleukin-2 production in lymphocytes in vitro by benzene and its metabolites, hydroquinone and p-benzoquinone.

The effects of benzene and its metabolites, hydroquinone and p-benzoquinone (PBQ) on RNA synthesis in mouse spleen lymphocytes in vitro were studied. Benzene and the quinones were shown to inhibit RNA synthesis in a dose-dependent manner at concentrations which had no significant effect on lymphocyte viability. Furthermore, 5 microM PBQ, the putative toxic metabolite of benzene, was shown to inhibit the formation of the T-cell growth factor IL-2. These results suggest that inhibition of RNA synthesis in lymphocytes by benzene may prevent the production of factors required for hemopoiesis and thus contribute to the aplastic anemia caused by benzene.

Animals↗
PubMed1985

The inhibition of mitochondrial DNA replication in vitro by the metabolites of benzene, hydroquinone and p-benzoquinone.

Rat liver mitochondria incubated with the metabolites of benzene, p-benzoquinone or 1,2,4-benzenetriol, showed a dose-dependent inhibition of [3H]dTTP incorporation into mtDNA with median inhibitory concentrations of 1 mM for each compound. Benzene and the metabolites phenol, catechol and hydroquinone did not inhibit at concentrations up to 10 mM. Similarly, incubation of p-benzoquinone or hydroquinone with rabbit bone marrow mitochondria showed a dose-dependent inhibition of mtDNA synthesis with 50% inhibition at 1 mM and 10 mM, respectively. That these metabolites inhibit mitochondrial replication was evidenced by the fact that [3H]dTTP incorporation into characteristic 38S, 27S and 7S mitochondrial replication intermediates was decreased by the quinones, as analyzed on 5-20% neutral sucrose velocity gradients. p-Benzoquinone, hydroquinone and 1,2,4-benzenetriol inhibited the activity of partially purified rat liver mtDNA polymerase gamma using either activated calf thymus DNA or poly(rA) X p(dT)12-18 as primer/template, with 50% inhibitory concentrations of 25 microM, 25 microM and 180 microM, respectively. Preincubation of the metabolites with polymerase gamma or primer/template, followed by removal of the unreacted metabolite by gel filtration, indicated that inhibition resulted from interaction of the metabolites with the enzyme, rather than with the template. Binding appeared to involve a sulfhydryl residue on the enzyme since the binding of [14C]hydroquinone was prevented by N-ethylmaleimide. The ability of hydroquinone or p-benzoquinone to inhibit binding of [14C]hydroquinone to the enzyme suggests that the compounds bind to a common site or are converted to a common intermediate. Inhibition of, or changes in, replication in mitochondria of bone marrow cells by hydroquinone and p-benzoquinone may explain the changes in the mitochondrial genome observed in marrow stem cells in acute myelogenous leukemia and may suggest a mechanism for benzene leukemogenesis.

Animals↗
PubMed1985

Assessment of calcium homeostasis in the critically ill surgical patient. The diagnostic pitfalls of the McLean-Hastings nomogram.

Hypocalcemia is a common problem in critically ill surgical patients. We prospectively evaluated whether measurement of the total serum calcium (Ca) concentration or calculation of the serum ionized Ca level (by the McLean-Hastings nomogram) accurately reflects the measured serum ionized Ca level. Although 71% and 58% of 156 predominantly surgical intensive care unit (ICU) patients were hypocalcemic by the total serum Ca or calculated ionized Ca level, respectively, only 12% were hypocalcemic by directly measured serum ionized Ca measurement. The total serum Ca and calculated ionized Ca concentrations were sensitive (95% and 89%, respectively) but lacked specificity (32% and 46%, respectively) in predicting ionized hypocalcemia. Analyses of Ca binding to albumin in the serum of surgical ICU patients and normal subjects suggested that there is a circulating factor in critically ill patients that increases the binding of Ca to albumin. These observations may explain why the McLean-Hastings nomogram underestimates the protein-induced changes in serum Ca in critically ill surgical subjects. We conclude that: total serum Ca and calculated ionized Ca concentrations are poor indicators of the true serum ionized Ca status in critically ill surgical patients, and we recommend direct measurement of serum ionized Ca levels in these patients; and variability in the affinity of Ca for binding proteins in critical illness may explain the poor correlation between serum total and ionized Ca measurements.

Blood Proteins↗
PubMed1984

Covalent binding of benzene and its metabolites to DNA in rabbit bone marrow mitochondria in vitro.

Rabbit bone marrow mitochondria, stripped of their outer membrane (mitoplasts), have been shown to carry out the NADPH-dependent bioactivation of radiolabelled benzene in vitro to metabolites capable of covalently binding to mtDNA, thereby inhibiting transcription. The metabolites of benzene produced in bone marrow cells by the microsomal cytochrome P-450 are thought to be phenol, catechol, hydroquinone and p-benzoquinone (Andrews et al., Life Sci., 25 (1979) 567; Irons et al., Chem.-Biol. Interact., 30 (1980) 241). Incubation of mitoplasts from rabbit bone marrow cells in vitro with varying concentrations of the putative microsomal metabolites showed a concentration-dependent inhibition of RNA synthesis. The 50% inhibitory molar concentration (IC50) for each metabolite was determined to be: 1,2,4- benzenetriol , 6.3 X 10(-7); p-benzo-quinone, 2 X 10(-6); phenol, 2.5 X 10(-5); hydroquinone, 5 X 10(-5); catechol, 2 X 10(-3); benzene, 1.6 X 10(-2). DNA, isolated from rabbit bone marrow cell or rat liver mitoplasts prelabelled in DNA with [3H]dGTP and exposed to [14C]benzene in vitro, was enzymatically hydrolyzed to nucleosides which were chromatographed on a Sephadex LH-20 column to separate free nucleosides from nucleoside-adducts. The elution profiles indicated that rat liver mtDNA contained six guanine nucleoside-adducts and rabbit bone marrow cell mtDNA contained seven guanine nucleoside-adducts. Incubation of bone marrow mitoplasts in vitro in the presence of benzene and the hydroxyl radical scavenger, mannitol, resulted in the inhibition of formation of four of the guanosine-adducts. When [3H]dATP was substituted as the prelabelled precursor nucleotide, the LH-20 column profile indicated that two adenine nucleoside-adducts were also formed from benzene in vitro. Furthermore, a comparison of the Sephadex LH-20 column profiles of purine adducts derived from [14C]benzene- and [3H]dGMP-labelled mtDNA with profiles generated by individually incubating each of the putative unlabelled metabolites with bone marrow mitoplasts in vitro has indicated that p-benzoquinone, phenol, hydroquinone and 1,2,4- benzenetriol form adducts with guanine. One of the two adenosine-adducts may arise from hydroquinone; the compound forming the other adduct is unknown at the present time. Exposure of mitoplasts to catechol in vitro resulted in the formation of a guanine nucleoside-adduct that was present in rat liver mtDNA but absent from the DNA isolated from rabbit bone marrow cell mitoplasts exposed to [14C]benzene in vitro. This suggests that catechol is probably not a major metabolite of benzene formed in bone marrow cell mitochondria.

Animals↗
PubMed1984

Inhibition of mRNA synthesis in rabbit bone marrow nuclei in vitro by quinone metabolites of benzene.

mRNA synthesis by rabbit bone marrow nuclei has been shown to be inhibited by the quinone metabolites of benzene, hydroquinone and p-benzoquinone, in a concentration-dependent manner with 50% inhibitory concentration (IC50[M]) for both compounds of 6 X 10(-6) M. Catechol and 1,2,4-benzenetriol also showed a concentration-dependent inhibition of synthesis, however, 50% inhibition was not reached by 10(-4) M. Phenol did not inhibit mRNA synthesis even at 10(-3) M. It is possible that myelotoxicity from benzene might result from such an inhibition of mRNA synthesis by quinone metabolites in pluripotent and/or committed bone marrow stem cells.

Animals↗
PubMed1984

Albumin coating of a knitted polyester arterial prosthesis: an alternative to preclotting.

Coating a knitted polyester arterial prosthesis with cross-linked albumin fills the interstices of the graft and relieves the surgeon of the necessity to preclot . This is of particular value in patients whose blood clotting properties are hypercoagulable, or hypocoagulable . In addition, such prostheses require less handling, which can lower the risk of bacteremic colonization and shorten the operative time. The in vivo behavior of the implanted albuminated prosthesis in the thoracic aorta of dogs is similar to that of preclotted grafts, although the sequences of early healing are different. The preclotted graft develops a continuous, thick thrombotic matrix on its luminal surface during the first 4 hours of implantation. Following the initiation of the fibrinolytic mechanism 24 to 48 hours postoperatively, this thrombotic deposit quickly recedes , leaving blood cells and platelets adhering here and there to the prosthetic surface. In comparison, the albuminated coating is not associated with major early thrombotic deposits. The albumin remains visible between the filaments during the first 2 weeks of implantation. Both treated and control grafts contain numerous thrombi on their inner surface after 1 to 2 weeks. After 1, 3, and 6 months, both implants are well encapsulated and present a glistening and continuous luminal surface. This excellent healing, however, can be compromised should the graft adhere too closely to the animal's lungs.+2

Albumins↗
PubMed1984

Correlation of benzene metabolism and histological lesions in rainbow trout (Salmo gairdneri).

Rainbow trout metabolized benzene to phenol and catechol, and excreted them primarily as glucuronide conjugates. Treatment with benzene or toluene resulted in spleenomegaly that may be a more generalized solvent effect on the circulating RBCs. Anemia was observed only in the benzene-treated animals. This may be a result of decreased erythropoiesis in the head kidney or a direct effect of the principal metabolite (phenol) or its reactive intermediate. Both the spleen and liver metabolized benzene in vitro, while no metabolism was observed in the head kidney or trunk kidney. This fact, along with the observed "covalently" bound material in the liver, spleen, and kidney in vivo, can be interpreted as indicating that a reactive metabolite is formed outside the head kidney and is transported to the head kidney where it results in the observed hematopoietic damage. A similar hypothesis has been suggested for the observed protection by partial hepatectomy against bone marrow effects in mammalian species. The effects observed in trout are similar to those reported for higher mammalian species, but the metabolic profile is simpler, and studies in these animals may shed light on the mechanism of benzene toxicity. The techniques developed for monitoring benzene toxicity in trout can also be used for examining other environmental pollutants that may affect the hematopoietic system of fish.

Animals↗
PubMed1984

The forensic pediatrician as a child advocate.

This paper focuses on the medicolegal activities of a group of six pediatricians from the Winnipeg Children's Hospital. Increasing legal demands resulted in 93 court appearances for the group, during a 12-month period ending in December 1982. The educational processes and the individual court room issues confronted by both the courts and the six pediatricians, dealing with problems of child protection, are described. Major pediatric roles and principles of advocacy resulting from these experiences are delineated. Society's accountability is discussed with specific reference to the pediatrician's role.

Adult↗