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Biomedical subjects

B Martin

Publications and source records attributed to B Martin.

At least 289 records · Page 16Linked to original sources
PubMed1989

The difficulty of cloning Streptococcus pneumoniae mal and ami loci in Escherichia coli: toxicity of malX and amiA gene products.

Stability problems are frequently encountered when cloning pneumococcal DNA in Escherichia coli multicopy plasmid vectors such as derivatives of ColE1. In this paper, we report our investigations of these problems using the pneumococcal mal and ami regions. We offer evidence that, in both cases, promoters are not the primary cause of cloning problems. Indeed, successful cloning of mal and ami promoters has been achieved with standard vectors (devoid of transcriptional terminators flanking the insertion site). Moreover, we show that the entire mal fragment can be introduced into an E. coli strain harboring a chromosomal mutation that reduces plasmid copy number. The cause of the cloning problems has been traced to the malX and amiA structural genes. Overexpression of these genes, which probably encode lipoproteins, could have deleterious effects on E. coli hosts, possibly as a result of impairing the protein export machinery.

Cloning, Molecular↗
PubMed1989

Curriculum evaluation.

Curriculum evaluation is an essential phase of curriculum development. Through evaluation a faculty discovers whether a curriculum is fulfilling its purpose and whether students are actually learning. The purpose of this manuscript is to describe the implementation of a formative evaluation used by faculty members of a baccalaureate nursing program. PAIREDAR, a three phase generic model, was utilised to conduct this formative evaluation. The definition and description of this model are included. The process used by the evaluation team within the framework of the model is described in a step by step fashion. Faculty and student interview question sets and a sample course evaluation worksheet are presented.

Curriculum↗
PubMed1989

N-terminal galanin-(1-16) fragment is an agonist at the hippocampal galanin receptor.

The galanin N-terminal fragment [galanin-(1-16)] has been prepared by solid-phase synthesis and by enzymic cleavage of galanin by endoproteinase Asp-N. This peptide fragment displaced 125I-labeled galanin in receptor autoradiography experiments on rat forebrain and spinal cord and in equilibrium binding experiments from high-affinity binding sites in the ventral hippocampus with an IC50 of approximately 3 nM. In tissue slices of the same brain area, galanin-(1-16), similarly to galanin, inhibited the muscarinic agonist-stimulated breakdown of inositol phospholipids. Upon intracerebroventricular administration, galanin-(1-16) (10 micrograms/15 microliters) also inhibited the scopolamine (0.3 mg/kg, s.c.)-evoked release of acetylcholine, as studied in vivo by microdialysis. Substitution of [L-Trp2] for [D-Trp2] resulted in a 500-fold loss in affinity as compared with galanin-(1-16). It is concluded that, in the ventral hippocampus, the N-terminal galanin fragment [galanin-(1-16)] is recognized by the galanin receptors controlling acetylcholine release and muscarinic agonist-stimulated inositol phospholipid breakdown as a high-affinity agonist and that amino acid residue [Trp2] plays an important role in the receptor-ligand interactions.

Animals↗
PubMed1989

Nucleotide sequence of the Streptococcus pneumoniae hexB mismatch repair gene: homology of HexB to MutL of Salmonella typhimurium and to PMS1 of Saccharomyces cerevisiae.

The Hex mismatch repair system of Streptococcus pneumoniae acts both during transformation (a recombination process that directly produces heteroduplex DNA) to correct donor strands and after DNA replication to remove misincorporated nucleotides. The hexB gene product is one of at least two proteins required for mismatch repair in this organism. The nucleotide sequence of a 2.7-kilobase segment from the S. pneumoniae chromosome that includes the 1.95-kilobase hexB gene was determined. The gene encodes a 73.5-kilodalton protein (649 residues). The spontaneous hex Rx chromosomal mutant allele with which a mutator phenotype has been associated is shown to result from a single base substitution (TAC to TAA) leading to a truncated HexB polypeptide (484 residues). The HexB protein is homologous to the MutL protein, which is required for methyl-directed mismatch repair in Salmonella typhimurium and Escherichia coli, and to the PMS1 gene product, which is likely to be involved in a mismatch correction system in Saccharomyces cerevisiae. The conservation of HexB-like proteins among procaryotic and eucaryotic organisms indicates that these proteins play an important common role in the repair process. This finding also suggests that the Hex, Mut, and PMS systems evolved from a common ancestor and that functionally similar mismatch repair systems could be widespread among procaryotic as well as eucaryotic organisms.

Amino Acid Sequence↗
PubMed1989

Apolipoprotein B metabolism in homozygous familial hypercholesterolemia.

This report describes the metabolism of apolipoprotein B-containing lipoproteins in seven familial hypercholesterolemic (FH) homozygotes and compares the results to the values obtained from five healthy control subjects. The concentration, composition, and metabolism of large, triglyceride-rich very low density lipoproteins (VLDL1, Sf 60-400) were the same in the control and FH groups, indicating that this component of the VLDL delipidation cascade ws unaffected by the absence of receptors. In contrast, familial hypercholesterolemic small VLDL2 (Sf 20-60) was enriched with cholesterol and depleted in triglyceride. Moreover, its plasma concentration was elevated as a result of an increase in its synthesis and a defect in the removal of a remnant population within this density interval. The latter accounted for up to 50% of the total mass of the fraction. Onward transfer of apolipoprotein B (apoB) from small VLDL through intermediate density lipoprotein (IDL) to low density lipoprotein (LDL) was retarded, suggesting that receptors were involved in this supposedly lipase-mediated event. IDL and LDL concentrations increased up to fourfold above normal in the plasma of the FH patients due partly to the delay in maturation and partly to defective direct catabolism. We conclude that the LDL receptor plays multiple and important roles in the metabolism and transformation of apoB-containing particles in the Sf 0-400 flotation interval.

Adult↗
PubMed1988

Purification and characterization of an endogenous protein modulator of radioligand binding to "peripheral-type" benzodiazepine receptors and dihydropyridine Ca2+-channel antagonist binding sites.

Acidified extracts of rat antral stomach chromatographed on octadecylsilane cartridges contained material that inhibited the binding of [3H]Ro 5-4864 (4'-chlorodiazepam) and [3H]nitrenidipine to "peripheral-type" benzodiazepine receptors and dihydropyridine Ca2+-channel antagonist binding sites respectively. This material reduced the apparent affinities of both radioligands without significantly affecting the maximum number of binding sites. In contrast, the binding of [3H]diazepam, [3H]Ro 15-1788 (ethyl-8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo [1,5a][1,4] benzodiazepine-3-carboxylate), and [3H]3-carbomethoxy-beta-carboline to "brain-type" benzodiazepine receptors and [3H]dihydroalprenolol binding to beta-adrenergic receptors were unaffected by this material. Subsequent column chromatography on hydroxylapatite purified this material by greater than 2000-fold. This semi-purified substance was resolved by reverse phase HPLC as one u.v. adsorbing peak that inhibited both [3H]Ro 5-4864 and [3H]nitrendipine binding. The activity of this 16,000 dalton substance was destroyed completely by both heat treatment and pronase and partially reduced by trypsin. Furthermore, the inhibitory activity of this substance was enhanced by Ca2+ in a concentration-dependent fashion (0.1 to 10 mM). Comparison of TLC scans of 2-9,10[3H]dipalmitoyl-phosphatidylcholine incubated with either the HPLC purified material or authentic phospholipase A2(PLA2) (Naja naja) revealed that this substance has enzymatic properties indistinguishable from PLA2. These findings suggest that this endogenous protein may be a PLA2 isoenzyme which may modify both "peripheral-type" benzodiazepine receptors and dihydropyridine Ca2+-channel antagonist binding sites.

Animals↗
PubMed1988

Simultaneous demonstration of glia- and glioma-associated antigens in human astrocytomas.

Glial fibrillary acidic protein (GFAP) and glioma-associated antigens (GAA) defined by monoclonal antibodies (MAbs) were demonstrated simultaneously in human astrocytoma tissue. GFAP was stained by PAP-method, GAA were visualized by avidin-biotin-technique using alkaline phosphatase. In primary and secondary tumors as well as in tissue culture heterogeneity of GFAP- and GAA-expression is obvious. GFAP is mostly restricted to cell processes and less marked in the perinuclear space. Depending on the individual antibody, MAbs-positive material is located either in the tumor cell plasma (MUC 8-22) or on cell surface membranes (MUC 2-63). There is remarkable expression of GAA in cell clusters which fail to express GFAP. At higher magnification, 3 types of cellular reactivity are detectable: (a) cells which react only with anti-GFAP, (b) cells which react only with anti-GAA and (c) cells which express both, GFAP and GAA, especially those of protoplasmic astrocyte type. These cells also occur in subcutaneous tumor grafts, and may thus represent not only a reactive event, but be part of tumor cell populations.

Animals↗
PubMed1988

Significance of scrotal calcifications detected by sonography.

A total of 468 patients were evaluated using a high-resolution scanner with a high-frequency transducer. Scrotal calcifications were diagnosed by sonographic criteria in 22 patients. Size, number, arrangement, and especially location (testicular or extratesticular) were precisely determined. Calcifications, located within testicular parenchyma, particularly if multiple and grouped, were considered highly suspicious for malignancy, and surgery was mandatory. Based on these sonographic features, 9 patients underwent surgery with pathological examination. In all, calcifications were confirmed and found in 4 necrotic germ-cell tumors, in a germ-cell tumor scar but with intratubular occult seminoma, in 1 epidermoid testicular cyst, in 1 tuberculous epididymo-orchitis, in 1 granulomatous orchitis, and at last, in 1 ruptured testis with an old hematoma. In the 13 remaining patients, calcification was usually defined as solitary, large, and extratesticular by sonography and considered benign. No changes were seen in follow-up examinations.

Adult↗
PubMed1988

Local anti-inflammatory activities of tixocortol 21-pivalate, inhibition of prostaglandins and leukotrienes synthesis, in carrageenin-induced pleurisy. Reversion of effects by RU 486.

Since a direct effect of tixocortol pivalate (TP) has been described on cyclooxygenase pathway, local anti-inflammatory activities of some 21 thiol derivatives of steroids were investigated on the carrageenin-induced pleurisy model in comparison with dexamethasone (Dex) or other anti-inflammatory drugs. LTC4/D4 contents in pleural fluid were assayed by RIA as well as PGE2 levels to characterize the effects on arachidonate pathways. After oral administration, TP was inactive up to 1 g/kg on exudate volume and leukocyte migration as expected for this strict local anti-inflammatory steroid contrary to Dex (ID50 = 0.05-0.41 mg/kg). When administered locally, TP and tixocortol (T) exerted a dose dependent inhibitory activity on exudate volume (ID30 = 12.4 micrograms or 13.1 micrograms/pleural cavity) and leukocyte count (ID30 = 83 or 230 micrograms); in the same conditions. Dex was more active (ID30 = 0.7 and 2.6 micrograms). All these steroids decreased PGE2 and LTC4/D4 contents in exudate fluids, respectively TP (50 micrograms/pleural cavity) by 28 and 63%; T (100 micrograms) by 33 and 31%; Dex (5 micrograms) by 43 and 40%. Local co-administration of RU 486 (50 micrograms) with either TP, T or Dex reversed the anti-inflammatory effects of all steroids, indicating in these conditions a local activity through glucosteroid receptor occupancy.

Administration, Topical↗
PubMed1988

Direct inhibition of cyclooxygenase pathway in platelets by tixocortol 21-pivalate: comparison with related structures, steroidal and non steroidal anti-inflammatory drugs.

Direct activity of the local anti-inflammatory steroid, tixocortol 21-pivalate (21 thioester derivative of cortisol) on metabolism of exogenous arachidonate by rabbit platelets was investigated in vitro. Tixocortol 21-pivalate inhibited generation of prostanoids from cyclooxygenase with an IC50 value of 19.6 microM without affecting the 12-lipoxygenase pathway. In this model, reference anti-inflammatory glucosteroids were ineffective, whereas indomethacin and aspirin inhibited the cyclooxygenase activity. Among tixocortol 21-pivalate related structures, tixocortol and its disulfide dimere or several other tixocortol esters exhibited a quite similar efficacy while S-methylation and subsequent S-oxydations of tixocortol abolished inhibitory activity. These results indicate that tixocortol 21-pivalate in contrast to other glucosteroids is able to act directly on cyclooxygenase pathway and that some specific chemical environment of the 21 thiol side chain are required for this inhibition.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1988

Long-term preservation of dried phosphofructokinase by sugars and sugar/zinc mixtures.

We have demonstrated that sugars and suger/zinc mixtures can be used to preserve the activity of dried phosphofructokinase (PFK) during long-term storage over CaSO4. After 9 weeks in the presence of either 200 mM sucrose or 200 mM trehalose little loss of PFK activity was noted, with almost 60% of the original prefreeze-dry activity recovered when samples were rehydrated. Even reducing sugars protected the dried enzyme throughout the entire storage period. Of the sugars tested, 200 mM lactose provided the most stability to PFK; at the end of the dry storage, over 80% of the initial activity was recovered. With either 200 mM maltose or 400 mM glucose, about 40% of the initial activity was recovered at the end of the experiment. With all the sugars tested, the addition of 0.6 mM Zn2+ to sugar/PFK mixtures enhanced the stability of the enzyme, and no long-term adverse effects of the metal ion on enzyme activity were noted.

Animals↗
PubMed1988

A preliminary behavioral investigation of PMMA, the 4-methoxy analog of methamphetamine.

The controlled-substance analog N-monomethyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA) may be viewed as being either the 4-methoxy analog of methamphetamine or the N-methyl analog of 1-(4-methoxyphenyl)-2-aminopropane (PMA). Because of its abuse potential, PMMA was examined with regard to (a) its stimulus properties in rats trained to discriminate either 1.0 mg/kg of (+)amphetamine or (+/-)DOM from saline, (b) its toxicity (isolated and aggregated) in mice relative to (+/-)PMA, and (c) its locomotor stimulant activity in mice relative to (+/-)amphetamine, (+/-)methamphetamine, and (+/-)PMA. Racemic PMMA produced neither DOM-like nor, unlike PMA, amphetamine-like stimulus effects. There was no significant difference between the 24-hr isolated (LD50 = 63 mg/kg) and aggregated (LD50 = 53 mg/kg) toxicity, and PMMA did not produce significant locomotor stimulation at doses of up to 30 mg/kg. The present results suggest that while PMMA may produce central effects it does not appear to behave as a simple amphetamine-like agent.

Amphetamine↗