Abundant Binding Sites Research Articles

Absence of Abundant Saturable Binding Sites for Halothane or Isoflurane in Rabbit Brain

We tested whether the existence of saturable binding sites for anesthetics causes the solubility of halothane or isoflurane in rabbit brain not to obey Henry's law. For each anesthetic, we measured brain/gas partition coefficients (paired samples) at approximately 0.05 MAC and 5 MAC at 38.5 degrees C. In addition, for halothane, brain/gas partition coefficients (paired samples) were determined at 0.05 MAC and 2 MAC. The values for halothane at 0.05 MAC, 2 MAC, and 5 MAC did not differ; values for isoflurane at 0.05 MAC and 5 MAC did not differ. Over the range of anesthetic partial pressures studied, no evidence for saturable binding was found. We conclude that the solubility of halothane and isoflurane in brain is independent of the partial pressure applied; inhaled anesthetics obey Henry's law.

Identification within the simian virus 40 genome of a chromosomal loop attachment site that contains topoisomerase II cleavage sites

We demonstrate that the simian virus 40 genome contains a single MAR (matrix association region) that maps within a large T-antigen coding region (nucleotides 4071 to 4377). This region contains topoisomerase II cleavage sites, exhibits sequence similarity with cellular MARs, and recognizes the same evolutionarily conserved, abundant nuclear binding sites seen by cellular MARs.

Neurotransmitters and modulators in the cerebellum

It is well-known that anatomically, the cerebellum has a very uniform and stereotyped structure with geometrical precision. The Purkinje cell, which provides the unique output from the cerebellum, receives two excitatory inputs: one is from parallel fibers, axons of the granule cells, and the other is from climbing fibers which originate at the inferior live. In order to characterize the neurotransmitter of the climbing fibers, the electrophysiological properties of Purkinje cells were first studied by intradendritic as well as intrasomatic recordings from Purkinje cells of guinea pig cerebellar slices. Comparison of the reversal properties, ionic dependency and the sensitivity to various antagonists of the climbing fiber responses with those of responses to L-glutamate and L-aspartate supports the previous suggestion based on the bio- and histochemical observations that L-aspartate is more likely than L-glutamate to be the neurotransmitter of the climbing fiber synapses. The Purkinje cell is also reported to contain a relatively high activity of NADP-linked prostaglandin (PG)D2 dehydrogenase and abundant PGD2 binding sites. PGs potentiate the actions of both excitatory and inhibitory neurotransmitter candidates in our experimental system. In this article, the role of prostaglandins in the neuronal transmission in the cerebellum is also described.

Stopped-flow kinetic studies on the interaction between echinomycin and DNA.

The kinetics of association between the quinoxaline antitumor antibiotic echinomycin and DNA have been studied by stopped-flow methods. With natural DNAs, the reaction profile is completely described by a single exponential, the time constant for which varies linearly with the DNA concentration. This bimolecular rate constant is similar for both calf thymus and Micrococcus lysodeikticus DNA (k = 6 X 10(4) M-1 s-1 at 25 degrees C, I = 0.01) and is probably dominated by interaction with relatively weak but abundant binding sites from which the antibiotic dissociates fairly quickly. The observed single exponential suggests a molecular mechanism of binding in which both chromophores of the antibiotic become intercalated simultaneously rather than sequentially; no transition from a mono-intercalated state to a bis-intercalated state could be detected. The reaction is slowed by a factor of about 3 on raising the salt concentration from I = 0.01 to I = 0.5. Binding to poly(dA-dT) is also described by a single exponential, the time constant for which is about 3 times faster than that seen with natural DNAs. By contrast, the interaction with poly-(dG-dC) requires two exponentials for a proper description, the faster of which is similar to that seen with natural DNAs. This may reflect the initial interaction of the antibiotic with two types of sequences, tentatively identified as GpC and CpG, from which it dissociates at very different rates. The differences in kinetic behavior may be explicable on the basis of an alternating B structure for poly(dA-dT) and a more classical B form for poly(dG-dC).

Germinal center B cells lack homing receptors necessary for normal lymphocyte recirculation.

Germinal center B cells (GCLC) are a discrete population of antigen-activated lymphoblasts that lack surface IgD and express abundant cell surface binding sites for peanut agglutinin (PNA). These phenotypic features render GCLC easily distinguishable from nearly all plasma cells, T cells, and unstimulated B cells, and have enabled us to identify and isolate GCLC from antigen-stimulated murine lymphoid organs. We have examined the migratory properties of these lymphoblasts in (a) short-term in vivo homing studies, and (b) an in vitro assay of lymphocyte binding to post-capillary, high endothelial venules (HEV) in frozen sections of Peyer's patches and peripheral lymph nodes. In the in vivo experiments, intravenously injected GCLC failed to migrate in significant numbers to peripheral lymphoid organs in comparison with T cells or IgD+ B cells. In the in vitro binding assay, GCLC did not adhere to HEV in either Peyer's patch or peripheral node sections. A variety of factors, such as preferential sequestration in the liver, may operate in vivo to influence the localization of these cells. However, their nearly total failure to migrate into lymphoid organs can best be explained by their inability to recognize and adhere to the specialized HEV which normally mediate the emigration of recirculating lymphocytes from the blood into these sites. The concept that GCLC fail to express functional homing receptors for HEV has been further supported by studies using MEL-14, a monoclonal antibody that appears to recognize the lymphocyte surface receptor for peripheral node HEV: In contrast to most peripheral lymphocytes, GCLC fail to bind MEL-14. These migratory and endothelial-recognition properties of GCLC, when viewed in the context of the possible role of these cells as precursors of plasma cells and/or memory B cells, have led us to propose that the inability of GCLC to recognize HEV may be transient and related to a phase of sessile B cell differentiation.

Chicken tissue binding sites for a purified chicken lectin.

A lactose-binding lectin previously purified from embryonic chicken muscle and adult chicken liver, and here referred to as chicken-lactose-lectin-I (CLL-I), was added to sections of various adult chicken tissues to detect available binding sites. Both the sites of binding of added CLL-I as well as the tissue distribution of endogenous CLL-I were determined by indirect immunofluorescence using a rabbit antibody to CLL-I followed by fluorescent goat anti-rabbit IgG. Some tissues such as intestine and kidney showed abundant extracellular binding sites for the lectin, primarily between cells, in basement membrane, and in material on the luminal surface. In contrast, adult heart showed no significant binding sites for CLL-I. Adult pancreas showed considerable endogenous CLL-I in an extracellular site surrounding exocrine lobules, but added CLL-I did not bind substantially. The distribution of CLL-I binding sites in intestine were mimicked by those of purpurin, another lactose-binding lectin. CLL-I binding sites were also detected on the surface of cultured chick embryo skin fibroblasts. The factors controlling the specific distribution of occupied and unoccupied CLL-I binding sites are not known.