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Biomedical subjects

G B Weiss

Publications and source records attributed to G B Weiss.

At least 109 records · Page 6Linked to original sources

Early idiopathic hemochromatosis with absent stainable bone marrow iron stores.

Early idiopathic hemochromatosis was diagnosed in a 52-year-old woman during the course of a hospitalization for another disorder. Quantitative studies revealed massive hepatic parenchymal siderosis and absent stainable marrow iron. Additionally, intestinal absorption and hepatic sequestration of radiolabeled iron were elevated. These studies in a patient with very early hemochromatosis, before organ damage, complement observations previously made in patients with advanced disease and provide further support for the concept that this disorder may result from impaired reticuloendothelial handling of iron.

Bloodletting↗

The problem of nuclease activity in nucleic acid hybridization reactions. Theoretical considerations.

The presence of nuclease during DNA-DNA or DNA-RNA hybridization reactions alters the kinetics of hybrid formation. Unfortunately, while the effect (even of small amounts of nucleas) on the Cot curve may be large, it may not be readily detectable. The effect of various types of nuclease is shown. As many nucleases cause a shift in the position rather than a change in shape of the curve, all studies involving nucleic acid hybridization should assay for the presence of nucleases and care must be taken to avoid their presence as contaminants.

DNA↗

Importance of full size complementary DNA in nucleic acid hybridization.

The size of the DNA product synthesized by RNA-directed DNA polymerase (isolated from avian myeloblastosis virus) was found to be important for complementary DNA (cDNA)-mRNA hybridization reactions. Incomplete cDNA to rabbit reticulocyte globin mRNA formed poor hybrids and presumably lacked sequences needed for hybridization. The size of the cDNA synthesized was influenced by the reaction conditions used. The complementary DNA product contained 10 S material when synthesis was done at high deoxynucleoside triphosphate concentrations (greater than 50 muM) while the product was smaller than the template when synthesis was at lower concentrations. The concentration and size (oligo(dT)6 to (dT)10) of primer had little or no effect on the product size. Increasing the concentration of 10 S globin mRNA caused the cDNA product to contain more small material. The cDNA synthesized at high deoxynucleoside triphosphate concentrations was fractionated into heavy, medium, and light fractions by alkaline sucrose density centrifugation. All hybridized to globin mRNA. The larger cDNAs had a higher TM when hybridized to globin mRNA, a lower dTMP/dCMP ratio (indicating that the poly(dT) region constituted a smaller fraction of the molecule), and gave increased protection of 125I-labeled mRNA from nuclease digestion. The full size cDNA was especially useful for studying the RNA transcribed from chromatin by RNA polymerase. The complement of the 5' end of the mRNA is contained only in full size cDNA; the 5' end is the part of the mRNA first transcribed by the RNA polymerase assuming correct transcription. Thus, full size cDNA can hybridize more effectively to the short RNA transcripts that are obtained than partial cDNA. RNA transcribed from rabbit bone marrow chromatin by Escherichia coli RNA polymerase hybridized twice as efficiently to complete cDNA as it did to partial cDNA demonstrating the usefulness of full size cDNA.

Animals↗

Transcription of single base oligonucleotides by ribonucleic acid-directed deoxyribonucleic acid polymerase.

The synthesis of DNA products complementary to artificial templates by the enzyme RNA-directed DNA polymerase isolated from avian myeloblastosis virus has been studied. Of the single base polyribonucleotides, poly (rC), poly(rA), and poly(rI) were active while poly (rG) and poly (rU) were almost inactive. The minimum length showing activity for an oligo (rC) template was 9; the minimum primer length of oligo(dG) was 3 or 4. In order to examine the fidelity of transcription, single base oligoribonucleotides of defined length were studied. Using (rC)13 as template and (dG)8as primer, the oligo (dG) product coelectrophoresed with the template. However, using (rA)-20 as template and (dT)10 as primer, a large (10-16s) product was formed. Similarly, using oligo (rI) (2.5S) as template and (dC)10 as primer, a large (greater than 22s) product was formed. No significant activity was obtained with oligo (rU) templates. RNA-directed DNA polymerase transcribes the various oligonucleotides differently: slippage with oligo (rA) and oligo (rI), faithful transcription with oligo (rC), and poor transcription with oligo (rU).

Avian Myeloblastosis Virus↗

Distribution of a lanthanide (147 Pm) in vascular smooth muscle.

In order to ascertain whether trivalent rare earth ions such as lanthanum (La+++) penetrate the cell membrane under physiological conditions, the extracellular and cellular distribution of promethium (147 Pm), a carrier-free rare earth radioisotope, was examined in rabbit aortic smooth muscle. As the duration of incubation was lengthened, uptake of 147Pm continued to increase; it was inhibited by La+++ and other rare earth ions (Nd+++, Lu+++) only when the 147 Pm/rare earth concentration ratio exceeded 1:10(6). However, equally high concentrations of Ca++ had no effect on 147Pm uptake. Efflux of 147Pm was only transiently increased by 1.5 mM La+++, and exposure to 0.05 mM EDTA elicited an increased 147Pm efflux with both transient and maintained components. The magnitude of the EDTA-induced increase in 147 Pm efflux was similar over a 30-fold range of EDTA concentration (0.05-1.5 mM); the limiting factor for 147Pm efflux is the rate of 147Pm desorption from the tissue rather than the extracellular concentration of EDTA. Loss of 147Pm in the presence of 0.05 mM EDTA could be described in terms of two specific washout components (the more rapid of which included 147Pm within the extracellular space and the slower of which had half-times of washout of approximately 7-10 minutes). Uptake of 147Pm was inhibited by lowering the incubation solution temperature to 0 degrees C or by procaine. However, concentrations of metabolic inhibitors (iodoacetate and dinitrophenol) which diminish loss of Ca++ from the cell did not decrease either the uptake or efflux of 147Pm. Thus, significant quantities of 147Pm do not appear to be accumulated within the cell or transported out of the cell; distribution of 147Pm can be most simply described in terms of a binding at and desorption from surface acessible fiber sites.

Animals↗

Alteration by phosphatidyl serine of tension responses and 45Ca distribution in aortic smooth muscle.

The effects of phosphatidyl serine (PS) on 45Ca distribution, 45Ca movements and contractions were examined in rabbit aortic smooth muscle. Contractile responses to submaximal concentrations of norepinephrine and histamine were potentiated by prior exposure to PS, but equivalent responses to potassium were unaffected. Addition of PS to the incubation solution decreased 45Ca uptake; exposure of aortic strips to PS during washout of either 45Ca or promethium (147Pm) resulted in maintained increases in efflux. These PS-induced alterations in net loss of 45Ca or 147Pm can be attributed to a decreased membrane reuptake and/or rebinding. However, the presence of PS during the washout significantly reduced the increases in 45Ca efflux rate elicited with either 0.05 mM concentrations of Ca++ or ethylenediamine tetraacetic acid. Thus, in rabbit aortic smooth muscle, exogenous PS can alter the availability and/or exchangeability of a membrane-bound Ca++ fraction. By specifically increasing the affinity for Ca++ at relevant membrane sites or stores. PS may enhance the ability of vascular smooth muscle to respond to stimulatory agents that mobilize Ca++ from these sites and, in this manner, potentiate contractile responses.

Animals↗

Effects of neomycin on 45Ca binding and distribution in canine arteries.

The effects of neomycin (7.0 mM) on 45Ca movements and distribution were investigated in canine aortae and in canine carotid and terminal mesenteric arteries. Uptake of 45Ca was measured in calcium-free solution; the 45Ca tissue spaces in the carotid and terminal mesenteric arteries were 2--4 times greater than those observed in the aorta. Exposure of the aortae and the terminal mesenteric arteries to 1.5 mM Ca++ during the washout elicited large increases in 45Ca efflux in both preparations (increase in terminal mesenteric greater than aorta). Moreover, in all three arterial preparations, neomycin reduced 45Ca uptake and induced a sustained increase in 45Ca efflux (effects on terminal mesenteric larger than or equal to carotid greater than aorta). The terminal mesenteric and carotid arteries may accumulate and bind 45Ca at superficial membrane sites (readily exchangeable 45Ca) to a greater degree than does the aorta. If Ca++ located at these membrane sites contributes directly to the maintenance of mechanical responsiveness, then agents which alter membrane binding of Ca++ (e.g. neomycin) may exert a stronger action on these highly reactive vessels. Thus, contractile responsiveness in peripheral arteries may depend upon depots of superficially bound Ca++ to a greater degree than in the more centrally located aorta.

Animals↗

Interactions between several rare earth ions and calcium ion in vascular smooth muscle.

The effects of lutetium (Lu+++), europium (Eu+++) and neodymium (Nd+++) on 45Ca distribution, 45Ca movements and contractions were examined in rabbit aortic smooth muscle. Tension responses to norepinephrine were markedly inhibited by all three rare earth ions; K+-induced responses and those to histamine were partially blocked. Addition of 1.5 mM Lu+++, Eu+++ or Nd+++ to the bathing solution reduced 45Ca uptake (Nd+++ greater than Eu+++ greater than greater than Lu+++), produced a maintained increase in 45Ca efflux from strips of media-intimal layer washed out in a 0-Ca++ solution (Nd+++ greater than Eu+++ greater than Lu+++), and, when present during 45Ca incubation (before 45Ca washout), altered the release of 45Ca elicited by addition of 1.5 mM Ca++ during the 45Ca washout (inhibition by Nd+++ greater than Eu+++ greater than Lu+++). The 3-fold greater uptake of 45Ca in a 0-Ca++ and 1.5 mM Lu+++ solution than in an equivalent Sr++ solution results from a decreased effect of Lu+++ upon the slower 45Ca washout component. The actions of Lu+++ were quantitatively similar to those of low La+++ concentrations (0.05-0.10 mM), whereas those of Nd+++ and Eu+++ resembled effects obtained with equimolar concentrations of La+++. Thus, the three rare earth ions produce qualitatively similar alterations in 45Ca uptake and superficial binding. The difference observed can be attributed to variations in their affinity for superficial Ca++ binding sites and in their ability to block Ca++ uptake.

Animals↗

Effects of metiamide on distribution of 14C-histamine in aortic smooth muscle.

The effects of metiamide on 14C-histamine movements and on contractions induced by histamine were studied in rabbit aortic smooth muscle. Low temperature (0 degrees C) and pretreatment with aminoguanidine or chloroquine markedly reduced the uptake of 14C. Component analysis of 14C efflux curves indicated that this decrease occurred mainly within the slower of two washout components. Addition of metiamide (but not pyrilamine) to the bathing solution decreased 14C-histamine uptake, and exposure to metiamide during the washout of 14C-histamine decreased the rate of efflux. These effects of metiamide on 14C-histamine movements can be explained in terms of an altered ability of rabbit aortic smooth muscle to accumulate histamine at some cellular site or store.

Animals↗

Ionic basis for intracellular 14C-nicotine accumulation in slices from different rat brain areas.

The effects of ionic alterations on the accumulation, distribution and movements of 14C-nicotine in slices from rat brain striatum, hypothalamus, cortex and cerebellum were studied. The uptake of 14C-nicotone is not dependent upon Na+ present in the incubation fluid because a K+-substituted (O-Na+) solution increased the 14C-nicotine tissue space, a tris-substituted (O-Na+) solution decreased the 14C-nicotine tissue space and a sucrose-substituted (O-Na+) solution did not change the amount of 14C-nicotine taken up when compared to the 14C-nicotine tissue space obtained in a normal incubation solution. However, all three Na+-free solutions elicited a sustained decrease in 14C-nicotine efflux. The increase in 14C-nicotine space produced in a K+-substituted (O-Na+) solution was present primarily in the slower component of a two-component washout, whereas the decrease produced in a tris-substituted (O-Na+) solution produced an equal percentage decrease in the size of both components. Most of the observed effects could be attributed to a linear relationship between the logarithm of the intracellular K+ concentration and the 14C-nicotine tissue space. In conclusion, it appears that there is an intracellular binding site for nicotine and that the degree of binding is dependent upon the concentration of K+.

Animals↗

Alteration of contractile function and calcium ion movements in vascular smooth muscle by gentamicin and other aminoglycoside antibiotics.

Experiments were conducted to examine the effects of certain aminoglycoside antibiotics on contractile responses and related calcium ion (Ca(2+)) movements in isolated vascular smooth muscle. Gentamicin, kanamycin, and streptomycin decreased contractile responses produced by norepinephrine, histamine, and high K(+) in rabbit aortic strips. The inhibitory action of these antibiotics on mechanical function was more pronounced when the Ca(2+) concentration of the bathing solution was decreased from 1.5 mM (normal Ca(2+) solution) to 0.05 mM (low Ca(2+) solution). The uptake of radiocalcium ((45)Ca) into the isolated media-intimal layer of rabbit aortae was decreased in a maintained manner by each antibiotic. With gentamicin, the inhibitory effect on (45)Ca uptake was shown to be dependent upon the concentration of gentamicin employed and to be more evident in a 0.1 mM Ca(2+) solution than in a normal Ca(2+) solution. In addition, the rate of (45)Ca efflux from the rabbit aortic media-intimal layer was increased in a sustained manner by gentamicin, streptomycin, and kanamycin. Furthermore, contractile responses induced by high K(+) and norepinephrine in canine carotid arterial strips were inhibited by gentamicin. Present findings indicate that aminoglycoside antibiotics interfere with Ca(2+)-linked events leading to activation of the contractile mechanism of vascular smooth muscle. These in vitro findings may partially explain the occurrence of in vivo cardiovascular depression that has occasionally been observed after the administration of chemically related antimicrobial agents.

Aminoglycosides↗