Biomedical subjects

H P Noteborn

Publications and source records attributed to H P Noteborn.

27 records · Page 2Linked to original sources

PubMed1987

The binding efficiency of polyclonal and monoclonal antibodies to DNA modified with benzo[a]pyrene diol epoxide is dependent on the level of modification. Implications for quantitation of benzo[a]pyrene-DNA adducts in vivo.

A number of polyclonal antibodies specific for DNA modified with (+/-)trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyre ne (BPDE) were obtained from the sera of New Zealand white rabbits immunized with BPDE-DNA, complexed with methylated bovine serum albumin (mBSA). Monoclonal antibodies were developed by fusion of mouse myeloma cells with spleen cells isolated from BALB/c mice immunized with the same complex of BPDE-DNA and mBSA. These antibodies have been characterized for specificity in a highly sensitive, enzyme-linked immunosorbent assay (ELISA). All antibodies showed a very high affinity for single-stranded BPDE-DNA, but had lower affinity towards native BPDE-DNA. The affinity for the free mononucleoside BPDE-dG was at least 100-fold lower than that for BPDE-DNA, and no affinity was detected for BP tetrols or DNA modified with N-acetoxy-N-acetyl-2-aminofluorene. A high cross reactivity was observed with DNA modified with (+/-)-trans-1,2-dihydroxy-anti-3,4-epoxy-1,2,3,4-tetrahydrochrysene++ +. Using five different antibodies, monoclonal or polyclonal, we observed that the antibody affinity for BPDE-DNA was dependent on the level of modification; in the competitive ELISA as little as 4 fmol BPDE-DNA (50 pmol/micrograms) was sufficient for 50% inhibition with our best antisera, but 17 fmol of the adduct was required when [3H]BPDE-DNA of low modification (1-10 fmol/micrograms) was used as inhibitor. When samples of [3H]BP-DNA isolated from the livers of mice, treated i.p. with different doses of [3H]BP were examined by competitive ELISA and calibrated with [3H]BPDE-DNA of low modification (1-10 fmol/micrograms), binding values calculated from the immunoassay were in good agreement with those obtained from radioactivity measurements. In contrast, when this DNA was quantitated in competitive ELISA using highly modified BPDE-DNA as standards, values by ELISA were 20-40% of those obtained by radioactivity. These results indicate that the use of serially diluted BPDE-DNA of high modification as standard competitor in the ELISA will lead to erroneous results in the measurement of adducts in DNAs modified to a low extent (biological samples). The property of antisera specific for BP-DNA, recognizing highly modified DNA more efficiently than DNA modified to a low extent, may be common to all antisera elicited against highly modified DNA immunogens. Therefore we conclude that antibody affinity must be tested also with DNA samples of low modification, obtained either in vitro or in vivo.

2-Acetylaminofluorene↗

PubMed1987

Ultrastructural demonstration of exocytosis in the pineal gland.

Granular vesicles are present in pinealocytes and in rudimentary photoreceptor cells of many vertebrates, sometimes in large amounts. Their dense cores have been shown to store proteinaceous compounds, but the way they are released remains speculative. The aim of this study was to demonstrate whether or not exocytosis is the mechanism by which secretory products stored within granular vesicles are released. Therefore, a method has been used allowing a clear ultrastructural study of secretory products by exocytosis, even in tissues in which this process of secretion is quite rare and/or very slow. Exocytotic figures have been clearly demonstrated in the three species studied: golden hamster, snake, and parakeet. Nevertheless, they were never commonly observed as it was the case in neurohypophysis, even in such animals as the parakeet and snake, in which granular vesicles are very numerous. The possible reasons of this observation are discussed.

Animals↗

PubMed1986

Ultrastructural demonstration of secretion by exocytosis in rat pinealocytes with the use of the tannic acid method.

In the rat pineal gland the mechanism of release of secretory material was studied ultrastructurally after incubating tissues in Ringer solution containing tannic acid. The results indicate that pinealocytes release the contents of secretory vesicles into the extracellular space via exocytosis, a phenomenon that has not been visualized previously in this cell type. This finding may reflect release of polypeptides by the pineal gland.

Animals↗

PubMed1985

The effect of ovine pineal compounds prepared under red or green light on the activity of male rat anterior pituitaries in vitro.

A high molecular weight fraction XM100R (MW A 100,000) was prepared by ultrafiltration from ovine pineals using two different extraction methods under red light conditions (lambda greater than 600 nm). This fraction stimulates the release of radioimmunologically active luteinizing hormone (LH) of anterior pituitaries in vitro. The ultrafiltration fraction PM30R (MW greater than 30,000 and less than 100,000) was found to be radioimmunologically active only when the "Bensinger" extraction procedure was applied. However, when comparable fractions were prepared under green light and incubated with half-pituitaries, all the incubation media of the ultrafiltrated fractions, XM100R, PM30R, PM10R (MW greater than 10,000 and less than 30,000) UM2R (MW greater than 1000 and less than 10,000), UM05R (MW greater than 500 and less than 1000) and UM05F (MW greater than 500), reacted with anti-LH. This may mean that under green light conditions the high molecular weight ovine pineal compounds in XM100R are disintegrated and/or split up into small molecules which can stimulate the release of LH, or crossreact with the anti-LH serum.

Animals↗

PubMed1982

Comparison of some peptidic and proteic ovine pineal fractions with a bovine pineal E5 fraction.

Using rather simple and mild extraction and separation methods, three ovine pineal fractions (XM 300 R-PP7.2' and PP7.2 S) were obtained, which contain peptidic/proteic substances and which show fluorescence characteristics of indoles. The ovine fractions were compared with the bovine pineal E-5 fraction. The ovine fractions are chemically sensitive to normal laboratory light and stable in red light (lambda greater than 600 nm). Immunologically, these fractions and the bovine E 5 fraction are stable. From the results of radioimmunological experiments it was concluded that the bovine pineal E 5 fraction as well as the ovine pineal fraction XM 300 R-PP7.2 and PP7.2S may contain (a) peptide(s) ending by the same carboxy terminal tripeptide Pro-Arg-Gly(NH2).

Animals↗

PubMed1981

Selective utilization of palmitoyl lysophosphatidylcholine in this synthesis of disaturated phosphatidylcholine in rat lung: a combined in vitro and in vivo approach.

1. The acyl-CoA:lysophosphatidylcholine acyltransferase system in rat lung microsomes was found to utilize selectively 1-[1-14C]palmitoyl-sn-glycero-3-phosphocholine when compared with 1-[9,10-3H2]stearoyl-sn-glycero-3-phosphocholine. This result was found with either palmitoyl-CoA, linoleoyl-CoA or an equimolar mixture of these acyl donors and confirms recent data reported by Holub, Piekarski and Possmayer (Can. J. Biochem. 58 (1980) 434-439). 2. The selective utilization of palmitoyl lysophosphatidylcholine from a mixture of lysophosphatidylcholine species may cause an increased isotopic ratio in phosphatidylcholine when compared with that of total lysophosphatidylcholine. Thus, when rats were injected with a single doubly labelled species, i.e. 1-[9,10-3H2]palmitoyl-sn-glycero-3-phospho[methyl-14C]choline, the isotopic ratio in both total and disaturated phosphatidylcholine from lung was nearly identical to that of the injected substrate. This suggested a direct acylation by lung acyl-CoA:lysophosphatidylcholine acyltransferases. By contrast, when a mixture of 1-[9,10-3H2]palmitoyl-sn-glycero-3-phospho[methyl-14C]choline and 1-stearoyl-sn-glycero-3-phospho[methyl-14C]choline was injected, the 3H/14C ratio in disaturated lung phosphatidylcholine increased to about 1.4-fold that of the injected substrate. 3. These data indicate that increased isotopic ratios in disaturated phosphatidylcholine of lung tissue, after intravenous injection of lysophosphatidylcholine, do not necessarily point to the involvement of lysophosphatidylcholine:lysophosphatidylcholine transacylase in disaturated phosphatidylcholine formation.

1-Acylglycerophosphocholine O-Acyltransferase↗

PubMed1979

Effects of sheep pineal fractions on the activity of male rat hypothalami in vitro.

High molecular weight substances could be isolated from sheep pineals with the "Bensinger" extraction method, followed by ultrafiltration of the waterlayer through different diaflomembranes. Two of the pineal fractions, XM100R and PM30R, stimulate the gonadotropin releasing activity of the medial basal hypothalamus (MBH). In experiments in which comparable pineal fractions were incubated without MBH and without pituitary and injected in immature mice no effect was detectable. All experiments in which a similar amount of rat cerebral cortex was used for incubation with pineal fractions did not show any activity.

Animals↗

PubMedDate not supplied

Presence of immunoreactive neurophysins of a higher molecular weight in addition to the 10.000 form in the ovine pineal gland.

Using an aqueous extraction followed by ultrafiltration through Amicon Diaflo membranes, two ovine pineal fractions were obtained, which contain immunoreactive neurophysin. The presence of neurophysin was monitored by radioimmunoassay, employing an antiserum raised against pituitary bovine neurophysin and selected because it reacts with neurophysins of many other mammals. From 50 g of wet ovine pineal glands 552 micrograms of immunoreactive neurophysins were obtained. About 5% of these immunoreactive neurophysins are eluted from three different Sephadex columns with an elution volume corresponding to Mr above 10,000 between bovine serum albumin and pituitary neurophysin. The remaining 95% of ovine immunoreactive pineal neurophysin (Mr 10,000) shares immunological and physico-chemical properties with highly purified bovine pituitary neurophysin used as a reference. From the results of gel filtration and affinity chromatography on LVP-Sepharose it was concluded that ovine pineal gland may contain a neurophysin precursor molecule in addition to the neurophysin Mr 10,000.

Animals↗

PubMedDate not supplied

Neurohypophyseal hormone-like peptides in the ovine pineal gland using reverse-phase liquid chromatography and radioimmunoassay.

A method is described for the determination of the neurohormone contents of ovine pineal tissue by radioimmunoassay (RIA) after successive fractionation on gel filtration in formic acid and reverse-phase liquid chromatography (HPLC). This method gives a good resolution for the neurohormones vasopressin, vasotocin and oxytocin, without a significant interference of aspecific cross-reacting of peptides with the RIA. An acid extract from ovine pineal tissue was found to contain amounts of immunoreactive AVP- and OXT-like peptides, whereas an AVT-like peptide was not detectable over background levels after HPLC with post-column RIA. It is concluded from our results that an AVT-like peptide is not present in ovine pineal tissue, and the pineal AVP- and OXT-like peptides appeared to be associated to neurophysin molecules.

Animals↗