Luteinizing hormone response to an intravenous infusion of substance P in normal men. The effect of synthetic substance P (SP), infused intravenously in doses of 0.5, 1.0, or 1.5 pmol/kg-1/min-1 for 60 minutes, on gonadotropin secretion was evaluated in seven healthy men. SP tests and a control test with normal saline were randomly performed at weekly intervals. During the tests, SP infusion did not produce untoward side effects or changes in blood pressure. Plasma testosterone concentrations were normal in all subjects and remained unmodified during all tests, regardless of the infused dose of SP. Plasma luteinizing hormone (LH) levels were not modified when either normal saline or the lowest dose of SP were infused, whereas they were significantly increased in a dose-dependent fashion when larger amounts of SP were administered. In contrast, plasma follicle-stimulating hormone (FSH) concentrations did not change significantly during any test. These data demonstrate for the first time in normal men that the systemic infusion of SP stimulates LH release, without modifications of FSH secretion.


Combination chemotherapy with cisplatin, vindesine and mitomycin-C for advanced, inoperable non-small-cell lung cancer. To assess the activity of chemotherapy with cisplatin, vindesine and mitomycin-C (PVM) in advanced non-small-cell lung cancer (NSCLC) and to test the feasibility of preemptive therapy with PVM. A phase II clinical trial of PVM in patients with NSCLC treated at the Royal Prince Alfred Hospital between June 1987 and July 1990. Forty-one patients with advanced, inoperable or recurrent NSCLC--22 women, 19 men, with a median age of 51 years. Thirteen patients had been treated previously with radiotherapy and/or surgery; 18 had extrathoracic metastases. Four patients previously deemed inoperable were treated preemptively with PVM before proceeding to radical surgery. Cisplatin 100 mg/m2, vindesine 5 mg and mitomycin-C 8 mg/m2, all given intravenously on Day 1, with vigorous hydration and antiemetic therapy. Cycles were repeated every four weeks. Objective tumour response to treatment, patient survival time, time to treatment failure, and treatment toxicity. There was one complete tumour response to PVM and 15 partial responses; 14 patients had stable disease and nine had progressive disease--yielding an objective response rate (complete plus partial responses) of 39% (16/41; 95% confidence interval [CI], 24%-56%). Responses were documented in all histological subgroups, in both locally advanced and disseminated disease, and in recurrent disease. Median survival of the group was six months (95% CI, 5-10 months; range, 0.5-19+ months), and is unchanged by exclusion of the four patients treated before surgery. Seven of the 41 patients (17%) survived 12 months or longer. Median time to treatment failure in patients who had an objective response was six months (95% CI, 5-10 months). Grade 3 or 4 nausea and vomiting occurred in 21 patients (51%). Haematological, renal and neurological toxicity were not major problems; there were no deaths from treatment toxicity. PVM is an active regimen in advanced NSCLC and can produce durable remissions. The potential palliative effects of PVM in incurable disease must be weighed against the risk of subjective toxicity.


Granulocyte colony stimulating factor in the management of chronic neutropenia. To report two cases of chronic neutropenic states successfully treated with granulocyte colony stimulating factor (G-CSF). A 23-year-old man with severe congenital cyclic neutropenia causing lifelong recurrent infections and consequent debilitation presented with intractable infected leg ulcers. A 56-year-old man with acquired idiopathic neutropenia presented with severe perianal infection and sepsis. Both patients were successfully treated with recombinant G-CSF. G-CSF is an agent with major clinical potential for the therapy of primary neutropenic states.


Cholinergic influence of K(+)-evoked release of endogenous histamine from rat hypothalamic slices in vitro. The effects of cholinergic agents on the K(+)-evoked release of endogenous histamine from hypothalamic slices of rats were examined by a superfusion method in vitro. Acetylcholine and carbamylcholine significantly inhibited K(+)-evoked release of histamine from the slices, and the effect of acetylcholine was antagonized by the muscarinic antagonist atropine. On the other hand, nicotine significantly enhanced the K(+)-evoked release and its effect was antagonized by the nicotinic antagonist hexamethonium. The effects of carbamylcholine and nicotine on histamine release from slices of whole hypothalamus and from slices of the anterior hypothalamic region, which do not contain cell bodies of the histaminergic neurons, were the same. Thus, the K(+)-evoked release of endogenous histamine from histaminergic fibers in the hypothalamic slices was inhibited by muscarinic agents and enhanced by nicotinic agents. These results suggest that muscarinic and nicotinic receptors exert antagonistic effects on the release of hypothalamic histamine from in vitro slice preparations. The cholinergic receptors modulating histamine release might be located presynaptically on the histaminergic terminals.


[Therapy of juvenile rheumatoid arthritis]. The correct drug treatment of JRA must consider the course and the subtype of the disease. Nonsteroidal antiinflammatory drugs are the first choice treatment, especially the recent ones which are more active and less toxic. The slow-acting antirheumatic drugs are the second choice treatment and must be employed in the chronically active stages of the disease; good results have been obtained with sulphasalazine and methotrexate both on clinical features and on blood biochemistry with relatively scarce side effects. Thymic hormones, cyclosporin A and intravenous immunoglobulins, though not yet widely experienced, can represent a worthwhile alternative to standard treatment in carefully selected cases. Steroids must be used only in special cases (particularly aggressive systemic JRA, carditis, severe anemia and those patients who fail to respond to usual treatments) and must be withdrawn as soon as possible to avoid adverse effects and steroid-addiction. Intraarticular long-acting steroids are the first choice treatment for rheumatoid monoarthritis.


CfT-I: an LTR-retrotransposon in Cladosporium fulvum, a fungal pathogen of tomato. A retrotransposon from the fungal tomato pathogen Cladosporium fulvum (syn. Fulvia fulva) has been isolated and characterised. It is 6968 bp in length and bounded by identical long terminal repeats of 427 bp; 5 bp target-site duplications were found. Putative first- and second-strand primer binding sites were identified. Three long open reading frames (ORFs) are predicted from the sequence. The first has homology to retroviral gag genes. The second includes sequences homologous to protease, reverse transcriptase, RNAse H and integrase, in that order. Sequence comparisons of the predicted ORFs indicate that this element is closely related to the gypsy class of LTR retrotransposons. Races of the pathogen exhibit polymorphisms in their complement of at least 25 copies of the sequence. Virus-like particles which co-sediment with reverse transcriptase activity were observed in homogenates of the fungus. This is the first report of an LTR retrotransposon in a filamentous fungus.


PMI40, an intron-containing gene required for early steps in yeast mannosylation. We have previously described a temperature-sensitive pmi40-1 mutant of Saccharomyces cerevisiae which is defective in glycosylation and secretion because of a thermolabile phosphomannose isomerase (PMI) activity. Inactivation of PMI at the restrictive temperature of 37 degrees C prevents synthesis of the GDP-mannose and dolichol-phosphate-mannose required for a number of critical mannosyl transfer reactions and results in cell death. Here, we report the isolation of the PMI40 gene by complementation of the corresponding mutation. The PMI40 gene contains an efficiently spliced intron which differs from the majority of those so far identified in S. cerevisiae in that it is short and the branch-forming structure has an AACTAAC motif replacing the highly conserved consensus TACTAAC. The 48.2-kDa protein predicted to be encoded by PMI40 contains amino acid sequences corresponding to those of internal peptides derived from purified S. cerevisiae PMI. Deletion of the PMI40 coding sequence results in a strain requiring D-mannose for growth. The PMI40 gene is located on chromosome V, and its transcription is increased 12-fold when cells are grown on D-mannose as sole carbon source instead of D-glucose. PMI enzyme activity, however, is not increased in D-mannose-grown cells, and PMI protein levels remain constant, suggesting that the PMI40 gene is subject to additional levels of regulation.


Analysis of clastogenic effect of Porto Alegre drinking water supplies on mouse bone marrow cells. Studies were conducted to evaluate the clastogenic activity of drinking water from Porto Alegre and Guaíba (Rio Grande do Sul, Brazil) estuarine waters. Mouse bone marrow was the target organ. C57B1/6 male and female mice received the water samples as their only liquid supply. Bone marrow cells were collected on the 16th day after the beginning of treatment. The analysis of metaphases demonstrated that the water supplies did not increase the structural chromosome aberration frequencies compared to the control groups. Concerning numerical alterations, only one treated female group showed a significant difference (loss of one chromosome) when compared to the control group, but this result is not considered relevant.


Requirement of mosXe protein kinase for meiotic maturation of Xenopus oocytes induced by a cdc2 mutant lacking regulatory phosphorylation sites. The p34cdc2 protein kinase is a component of maturation-promoting factor, the master regulator of the cell cycle in all eukaryotes. The activity of p34cdc2 is itself tightly regulated by phosphorylation and dephosphorylation. Predicted regulatory phosphorylation sites of Xenopus p34cdc2 were mutated in vitro, and in vitro-transcribed RNAs were injected into Xenopus oocytes. The cdc2 single mutants Thr-14----Ala and Tyr-15----Phe did not induce germinal vesicle breakdown (BVBD) upon microinjection into oocytes. In contrast, the cdc2 double mutant Ala-14/Phe-15 did induce GVBD. Both the Ala-14 and Ala-14/Phe-15p34cdc2 mutants were shown to coimmunoprecipitate cyclin B1 and to phosphorylate histone H1 in immune complex kinase assays. Microinjection of antisense oligonucleotides to c-mosXe was used to demonstrate the role of mos protein synthesis in the induction of GVBD by the Ala-14/Phe-15 cdc2 mutant. Thr-161 was also mutated. p34cdc2 single mutants Ala-161 and Glu-161 and triple mutants Ala-14/Phe-15/Ala-161 and Ala-14/Phe-15/Glu-161 failed to induce GVBD in oocytes and showed a decreased binding to cyclin B1 in coimmunoprecipitations. Each of the cdc2 mutants was also assayed by coinjection with cyclin B1 or c-mosXe RNA into oocytes. Several of the cdc2 mutants were found to affect the kinetics of cyclin B1 and/or mos-induced GVBD upon coinjection, although none affected the rate of progesterone-induced maturation. We demonstrate here the significance of Thr-14, Tyr-15, and Thr-161 of p34cdc2 in Xenopus oocyte maturation. In addition, these results suggest a regulatory role for mosXe in induction of oocyte maturation by the cdc2 mutant Ala-14/Phe-15.


Passive smoking and sister-chromatid exchanges in lymphocytes. The object of this study was to determine whether exposure to environmental tobacco smoke is associated with DNA damage reflected by the frequency of sister-chromatid exchange (SCE) in lymphocytes. Within a cross-sectional design, 106 male non-smoking adults, employees of two administrative companies, were divided on the basis of self-reported exposure into high and low passive smoking groups. The high exposed subjects (passive smokers, n = 50) lived with smokers, worked with smokers and were exposed to tobacco smoke for an average of 70 h/week. The low exposed non-smokers (n = 56) were exposed for an average of 5 h/week. Plasma cotinine levels for the passive smokers ranged between 0.4 and 9.0 ng/ml (median 1.4 ng/ml), and for the low exposed group between 0.0 and 1.9 ng/ml (median 0.4 ng/ml) (p less than 0.0001; Mann-Whitney test). No difference was observed between the two groups in the frequency of SCEs in lymphocytes: 4.66 +/- 0.05 for passive smokers and 4.68 +/- 0.04 for low exposed non-smokers (mean +/- SEM) (p = 0.80; t-test). Reclassification of subjects on the basis of plasma cotinine levels did not change the results substantially. These results are in accordance with observations that the increase in cancer risk due to passive smoking is small in comparison with the increase due to active smoking. The SCE test may be too insensitive to be useful for the evaluation of possible cytogenetic effects related to passive smoking.


Effects of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) on somatic mutation in a soybean test system. The mutagenic activity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) was assayed in heterozygous soybean plants (Y11y11), based on the appearance of mutational spots (yellow, dark green and twin) on the leaves. When soybean seeds were treated with IQ at concentrations of 0.01-0.1 microgram/ml, the frequency of mutational spots per leaf increased significantly in proportion to the concentration of IQ. At higher concentrations IQ was toxic. The mutagenicity of IQ was enhanced by pretreatment with the hepatic S9 fraction from Aroclor-induced rats. The numbers of yellow and dark green spots per leaf increased markedly by the treatment with IQ and S9-activated IQ, but the number of twin spots did not increase.


Genotoxic action of an aqueous extract of Heliotropium curassavicum var. argentinum. Heliotropium curassavicum var. argentinum is widely employed in gout, rheumatism, neuralgias, arteriosclerotic disorders, muscular algias, phlebitis, varix and other illnesses. In order to analyze the genotoxic effect produced in vitro by this medicinal plant, chromosomal aberrations (CA), mitotic index (MI) and anaphase delay (AD) were studied in the CHO cell line, with and without the addition of S9 mix. Prepared according to the Argentine pharmacopeia 0.1, 1, 10 and 100 micrograms/ml plant decoction (aqueous extract) were assayed. One hundred cells per culture were studied for CA and AD, while MI was calculated for 2000 nuclei. The results revealed a significant increase in the percentage of abnormal metaphases (p less than 0.001) and in total aberrations (p less than 0.001). Both the MI and the AD affected the cell cycle. All results were enhanced by the addition of an S9 fraction. The toxic effect could be associated with pyrrolizidine alkaloids and their N-oxides, which through a process of in vitro metabolism become activated by microsomal oxidation and change into pyrrolic derivatives.


Mutagenic potential of toluidine blue evaluated in the Ames test. Toluidine blue is a vital, metachromatic thiazine dye which is used as an adjunct in clinical examination for the early detection of asymptomatic recurrent or secondary primary carcinoma in individuals who are at high risk for developing oral cancer. Because available data on the mutagenicity of toluidine blue was limited and contradictory, this study was conducted to evaluate the mutagenic potential of toluidine blue in the in vitro Ames Salmonella test. Tester strains TA97a, TA98, TA100 and TA102 were used. Toluidine blue was tested at concentrations of 0.1, 1.0, 10, 50, 100, 250 and 500 micrograms/plate, with and without S9 microsomal activation, and positive and negative controls were included. Results from tests without S9 showed a significant increase (p less than 0.05) in number of revertants in TA102 and in TA97a with 50 and 100 micrograms toluidine blue/plate, respectively. In tests with S9 activation, doses of toluidine blue ranging from 10 to 250 micrograms/plate induced dose-related increases in the number of revertants in all 4 strains. The results of this study indicate that toluidine blue has a mutagenic effect in the Ames test.


Sister-chromatid exchange induction produced by in vivo and in vitro exposure to alpha-asarone. The effect of alpha-asarone, a chemical with hypocholesterolemic properties extracted from Guatteria gaumeri, on SCE induction was studied both in human lymphocytes in vitro and in murine bone marrow cells in vivo. A slight but consistent increase in SCE was observed in both biological systems.


Mutagenicity of N,N'-dimethylurea and methylamine hydrochloride in the Ames Salmonella/microsome test: absence of mutagenic response. Methyl isocyanate (MIC) in aqueous solution forms methylamine (MA) and N,N'-dimethylurea (DMU). MA in buffered system further converts into its salt form, methylamine hydrochloride (MAH). Therefore, MAH and DMU were evaluated for their mutagenic activity in the in vitro Ames Salmonella/microsome mutagenicity test. The liquid preincubation protocol was followed, using tester strains TA98, TA100 and TA104 of Salmonella typhimurium, in the presence of 0, 5, 15 and 30% Aroclor 1254-induced rat liver S9 mixture. DMU and MAH did not induce a mutagenic response in any of the tester strains, both in the presence and in the absence of S9 mixture. The results therefore confirm that MIC in its native form or as its unknown metabolites is responsible for the mutagenic activity reported earlier by us in the his tester strains TA100 and TA104 of Salmonella typhimurium (Mutation Res., 204 (1988) 123-129) and not due to its hydrolysis products, MA or DMU.


Suppressing effect of vanillin on chromosome aberrations that occur spontaneously or are induced by mitomycin C in the germ cell line of Drosophila melanogaster. In order to investigate the anticlastogenic effect of vanillin on ring-X loss, D. melanogaster females exposed to different vanillin concentrations were crossed with non-treated, MMC- or MMS-treated males. The results obtained with this in vivo investigation showed a significant inhibition of vanillin in the frequencies of spontaneous ring-X loss--59, 56, 38 and 36%--at the different concentrations used. In addition, vanillin treatment caused a significant suppression of MMC-induced ring-X loss. This decrease was observed only in the first 3 days after the interruption of vanillin treatment and at the concentrations of 0.5 and 1% of this flavoring agent. In contrast, vanillin did not show any effect on chromosome loss provoked by MMS. Therefore, the ring-X loss-decreasing effect of vanillin seemed to depend on the quality of DNA lesions and consequently on a specific enzymatic repair process present in the oocytes of D. melanogaster.


Modulation of the mutagenicity of three dinitropyrene isomers in vitro by rat-liver S9, cytosolic, and microsomal fractions following chronic ethanol ingestion. The effects of chronic ethanol feeding of rats on the ability of liver fractions to modulate the bacterial mutagenicity of three dinitropyrene isomers (1,3-, 1,6- and 1,8-DNP), which require bacterial enzymes but not an exogenous enzyme source for activation, were studied. The mutagenicity of the DNP isomers toward S. typhimurium TA98 and TA100 was attenuated in the presence of post-mitochondrial supernatants (S9) from both ethanol-fed and pair-fed rats albeit, that from the ethanol-fed group was more efficient in lowering the mutagenicity. The cytosolic fraction from ethanol-fed rats enhanced the mutagenicity of all of the DNP isomers in TA100. The most notable enhancement was with 1,3-DNP in which a more than 4-fold enhancement was obtained. Cytosol from pair-fed rats enhanced only the mutagenicity of 1,3-DNP, this by 2.9-fold. Cytosolic NADPH-nitroreductase activity from ethanol-treated rats toward 1,6-, 1,8- and 1,3-DNP was increased 2.8-, 1.7- and 1.3-fold, respectively over pair-fed controls. Cytosolic NADH-nitroreductase from ethanol-fed rats was increased with 1,3-DNP (1.7-fold) and 1,8-DNP (1.4-fold) as substrates, but not with 1,6-DNP. Microsomes decreased the mutagenicity of DNP similarly to S9, i.e., fractions from ethanol-fed rats were more efficient than those of pair-fed rats in deactivating all the DNP isomers. Per mg of protein, detoxification of DNP by S9 was more efficient than with microsomes, thus both cytosolic and microsomal enzymes are required for maximal detoxification. In summary, ethanol feeding modulates both the augmented cytosolic activation of DNP to mutagens and the deactivation of the direct-acting mutagenicity of DNP by microsomes. In combination, as is the case with S9, the microsomal detoxifying activity outcompetes the cytosolic activation.


A Babesia bovis 225-kilodalton protein located on the cytoplasmic side of the erythrocyte membrane has sequence similarity with a region of glycogen phosphorylase. A Babesia bovis gene sequence is described which encodes a geographically conserved epitope (recognized by monoclonal antibody (mAb) 23.8.34) of a 225-kDa protein located on the surface of merozoites and associated with the infected erythrocyte membrane. The gene sequence, derived from both genomic and cDNA copies, is 2044 bp long and has one long open reading frame encoding about one third of the 225-kDa protein. The open reading frame is expressed in an approximately 6,400 nucleotide RNA transcript. A 73-amino acid sequence occurs as 4 complete and 1 partial tandem repeats at the carboxy terminus of the partial protein sequence. The epitope recognized by mAb 23.8.34 was localized to the repeat region. Based on epitope localization with mAb 23.8.34, the repeat was exposed on the surface of merozoites and located near the cytoplasmic face of the erythrocyte membrane. The amino terminus of the protein was non-repetitive and had 21% identity (60% similarity) to glycogen phosphorylase over a region of 151 amino acids. In addition, the corresponding 5' DNA sequence hybridized to as many as 8 restriction fragments on Southern blots of genomic DNA. In contrast, the DNA sequence of the repeat hybridized to a single fragment. Both the repeat and multiple non-repeat DNA sequences were detected in a different geographic strain of B. bovis. These results indicate that the 5' end of the 225-kDa protein gene is related to a larger gene family, independent of the 3' end of the gene encoding the repeat.


Identification of the uncultured bacillus of Whipple's disease. Whipple's disease is a systemic disorder known for 85 years to be associated with an uncultured, and therefore unidentified, bacillus. We used a molecular genetic approach to identify this organism. The bacterial 16S ribosomal RNA (rRNA) sequence was amplified directly from tissues of five unrelated patients with Whipple's disease by means of the polymerase chain reaction, first with broad-range primers and then with specific primers. We determined and analyzed the nucleotide sequence of the amplification products. A unique 1321-base bacterial 16S rRNA sequence was amplified from duodenal tissue of one patient. This sequence indicated the presence of a previously uncharacterized organism. We then detected this sequence in tissues from all 5 patients with Whipple's disease, but in none of those from 10 patients without the disorder. According to phylogenetic analysis, this bacterium is a gram-positive actinomycete that is not closely related to any known genus. We have identified the uncultured bacillus associated with Whipple's disease. The phylogenetic relations of this bacterium, its distinct morphologic characteristics, and the unusual features of the disease are sufficient grounds for naming this bacillus Tropheryma whippelii gen. nov. sp. nov. Our findings also provide a basis for a specific diagnostic test for this organism.


A bovine albumin peptide as a possible trigger of insulin-dependent diabetes mellitus. Cow's milk has been implicated as a possible trigger of the autoimmune response that destroys pancreatic beta cells in genetically susceptible hosts, thus causing diabetes mellitus. Studies in animals have suggested that bovine serum albumin (BSA) is the milk protein responsible, and an albumin peptide containing 17 amino acids (ABBOS) may be the reactive epitope. Antibodies to this peptide react with p69, a beta-cell surface protein that may represent the target antigen for milk-induced beta-cell--specific immunity. We used immunoassays and Western blot analysis to analyze anti-BSA antibodies in the serum of 142 children with insulin-dependent diabetes mellitus, 79 healthy children, and 300 adult blood donors. Anti-ABBOS antibodies were measured in 44 diabetic patients at the time of diagnosis, three to four months later, and one to two years later. All the diabetic patients had elevated serum concentrations of IgG anti-BSA antibodies (but not of antibodies to other milk proteins), the bulk of which were specific for ABBOS: The mean (+/- SE) concentration was 8.5 +/- 0.2 kilofluorescence units (kfU) per microliter, as compared with 1.3 +/- 0.1 kfU per microliter in the healthy children. IgA antibodies were elevated as well, but not IgM antibodies. The antibody concentrations declined after diagnosis, reaching normal levels in most patients within one to two years. The initial decline involved anti-ABBOS--specific antibodies almost exclusively. Much lower serum concentrations of anti-BSA antibodies were found in all 379 control subjects, but only 2.5 percent of them had small amounts of ABBOS-specific IgG. Patients with insulin-dependent diabetes mellitus have immunity to cow's-milk albumin, with antibodies to an albumin peptide that are capable of reacting with a beta-cell--specific surface protein. Such antibodies could participate in the development of islet dysfunction.


Vasorelaxing effect in rat thoracic aorta caused by fraxinellone and dictamine isolated from the Chinese herb Dictamnus dasycarpus Turcz: comparison with cromakalim and Ca2+ channel blockers. The components of Dictamnus dasycarpus Turcz were tested for their vasorelaxing effect on the rat aorta, and fraxinellone and dictamine were shown to be effective vasorelaxants. In high K+ (60 mmol/l) medium, Ca2+ (0.03 to 3 mmol/l)-induced vasoconstriction was inhibited concentration-dependently by both agents. The IC50 for fraxinellone and dictamine were calculated to be about 25 mumol/l and 15 mumol/l (for Ca2+ concentration of 1 mmol/l), respectively. Cromakalim (0.2-10 mumol/l) relaxed aortic rings precontracted with 15 but not 60 mmol/l of K+. Fraxinellone and verapamil were more potent and effective in producing relaxation in 60 mmol/l than in 15 mmol/l K(+)-induced contraction. However, dictamine was more potent in producing relaxation in 15 mmol/l K(+)-induced contraction. Nifedipine (1 mumol/l), dictamine (100 mumol/l) and fraxinellone (100 mumol/l) relaxed the aortic contraction caused by KCl or Bay K 8644. The tonic contraction elicited by noradrenaline (NA, 3 mumol/l) was also relaxed by dictamine (500 mumol/l), but not by fraxinellone (500 mumol/l) in the nifedipine (1 mumol/l)-treated aorta. This relaxing effect of dictamine persisted in endothelium-denuded aorta. Glibenclamide (10 mumol/l) shifted the concentration-relaxation curve of cromakalim, but not that of dictamine, to the right in rat aortic rings precontracted with NA. Dictamine (500 mumol/l) did not affect tonic contraction of NA which are reduced by H-7 (1 mumol/l) in Ca(2+)-depleted medium. In conclusion, fraxinellone is a selective blocker of voltage-dependent Ca2+ channel, while dictamine relaxed the rat aorta by suppressing the Ca2+ influx through both voltage-dependent and receptor-operated Ca2+ channels.


Isolation and characterization of two novel peptide amides originating from myelin basic protein in bovine brain. During a systematic search for peptides that possess the C-terminal amide structure, two novel peptide amides, one with a tyrosine amide and the other with an alanine amide were isolated from bovine brain by acid extraction and sequential steps of reversed phase HPLC. Microsequence, amino acid and mass spectral analyses revealed the structures: Ac-Ala-Ala-Gln-Lys-Arg-Pro-Ser-Gln-Arg-Ser-Lys-Tyr-amide and Ac-Ala-Ala-Gln-Lys-Arg-Pro-Ser-Gln-Arg-Ser-Lys-Tyr-Leu-Ala-Ser-Ala-amide . These 12 and 16 residues peptides had the primary structure identical to the N-terminal fragment of myelin basic protein (MBP). The peptides were therefore designated myelin peptide amide-12 (MPA-12) and -16 (MPA-16). Unlike other amidated peptides, MPA might be generated from MBP by hydroxyl radicals produced via a Fenton reaction in situ. However, this unique amidation seems to occur exclusively to MBP in a site specific manner in the brain.


Neurochemical and histological analysis of motor dysfunction observed in rats with methylnitrosourea-induced experimental cerebellar hypoplasia. Histological and neurochemical changes related to motor dysfunction observed in rats after neonatal treatment with nitrosoureas were examined. Neonatal rats received subcutaneous injections of methylnitrosourea (MNU: 0.125 mmol/kg, s.c.) or ethylnitrosourea (ENU: 0.25 mmol/kg, s.c.) daily at 4,5,6 and 7 days post partum, a period of cerebellar granule cell, stellate cell and basket cell formation. At 14 days and 45 days after birth, MNU-treated rats displayed a lowering in motor coordination skills measured by tests of retainment ability on a rod of 26 mm diameter, chinning-climbing ability on parallel rods or retainment ability on a rotating rod. Histological examination at 14 days after birth showed a cerebellar hypoplasia with reduced cellularity of the internal granule cell layer and a disperse disposition of Purkinje cells in the granule cell layer. Cerebellar growth and cerebellar content and concentration of DNA were remarkably reduced in the MNU-treated rat. The degree of the reduction in cerebellar content of glutamic acid paralleled the degree of the cerebellar hypoplasia at 14 and 45 days after birth. In contrast, the concentrations of gamma-aminobutyric acid, acetylcholine, 5-hydroxytryptamine and norepinephrine were significantly increased by MNU treatment. ENU treatment control did not exert any significant changes in the neurotransmitters and motor coordination. These results suggest that the motor dysfunctions observed in MNU treated rats are induced by unbalanced output activities from Purkinje cells to motor neurons.


Tumor-specific binding of radiolabeled G-22 monoclonal antibody in glioma patients. Iodine-131-labeled G-22 monoclonal antibody F(ab')2 fragment reaching specifically with a glioma-associated surface glycoprotein was administered to 12 glioma patients to investigate its use in radioimaging of intracranial gliomas. No immediate or delayed side effects were attributable to antibody injection. Nine patients received the radiolabeled complex intravenously. The images of low-grade gliomas were generally poor and disappeared within 4 days. High-contrast images were obtained beyond the 7th day in high-grade gliomas except one case in the pineal region. Three patients received intraventricular or intratumoral administration. Clear images of all tumors were demonstrated from the 2nd until later than the 7th day. One patient with cerebrospinal fluid (CSF) dissemination of brainstem glioma demonstrated negative CSF cytology after intraventricular administration.


Brain tumors manifesting as intracranial hemorrhage. The clinical course and computed tomographic (CT) findings of 23 patients with brain tumors manifesting as tumoral hemorrhage were reviewed. The most common symptoms were headache and clouding of consciousness. A CT finding of a lesion located next to a solid or irregular clot indicated intratumoral hemorrhage. Precontrast CT demonstrating an indent on the hematoma surface was a valuable indicator of tumoral hemorrhage. A CT finding of accumulated levels of blood/fluid or a hyperdense mass containing small hematoma indicated intratumoral hemorrhage, and obscure hyperdensity indicated intratumoral hemorrhagic infarction. Such findings were often difficult to distinguish from spontaneous intracerebral hemorrhage due to other factors. The incidence of rebleeding from residual tumors was high, carrying a very poor prognosis, so radical removal of brain tumors with hemorrhage is very important.


Partial resection of the gyrus rectus in pterional approach to anterior communicating artery aneurysms. The effect of partial resection of the gyrus rectus during the unilateral pterional approach on surgical outcome was evaluated in 194 consecutive patients with ruptured anterior communicating artery aneurysms. Resection was performed more frequently in cases with poor clinical grade, in the acute stage, with superiorly directed aneurysms, and with high-positioned aneurysms. The surgical results were graded into three stages, and the follow-up results into five stages using the Glasgow Outcome Scale. Outcomes for 52 patients receiving gyrus rectus resection were compared with those for 142 patients without resection. There were no apparent effects caused by gyrus rectus resection on outcome.


PTA of supra-aortic arteries with temporary balloon occlusion to avoid distal embolism. Percutaneous transluminal angioplasty (PTA) was carried out in eight patients with cervical arterial stenosis; six in the subclavian and brachiocephalic arteries (5 with subclavian steal syndrome), one in the common carotid artery, and one in both the brachiocephalic and common carotid arteries (with subclavian steal syndrome). The PTA balloon catheters were introduced via the femoral artery in seven and brachial artery in one. To prevent distal embolization through the vertebral and internal carotid arteries, the blood flow in these vessels was temporarily occluded with a balloon catheter. The dilation of the stenotic areas was generally satisfactory. Antegrade blood flow was promptly obtained in the vertebral artery even in patients with subclavian steal syndrome. In all patients, the clinical symptoms improved. Two patients underwent repeat PTA because of restenosis.


Long-term follow-up of cerebral blood flow in patients with ruptured cerebral aneurysm. The xenon-133 inhalation technique was used to make three measurements of regional cerebral blood flow (CBF) in 34 patients with ruptured cerebral aneurysm: in the acute period (less than 14 days) after subarachnoid hemorrhage, in the subacute period (15-30 days), and in the chronic period (12-24 months). The hemispheric mean value of initial slope index was used as the mean CBF. The clinical outcomes were classified into good recovery (GR) (24 cases), moderate disability (MD) (5), and severe disability (SD) (5) on the Glasgow Outcome Scale. In all periods, the mean CBF significantly correlated with the outcome. GR patients had the highest mean CBF, MD patients the intermediate mean CBF, and SD patients the lowest mean CBF. GR patients had a near-normal mean CBF by the chronic period, while SD patients showed no significant CBF recovery throughout the course.


Absence of response to hypothalamic stimulation test in brain death. Immunoreactive corticotropin-releasing hormone (CRH) and growth hormone-releasing hormone (GHRH) are present in the plasma of the brain dead patients. These hypothalamic hormones may reflect some residual brain function after brain death. To examine the hypothalamic function, insulin-induced hypoglycemia and arginine infusion were performed in brain dead patients. Plasma CRH and GHRH were present initially, but levels did not increase significantly for 120 minutes after insulin injection. GH, adrenocorticotropic hormone, and cortisol levels did not increase either. Arginine load did not induce GH. These results suggest that hypothalamic hormones in the plasma after whole brain death do not reflect hypothalamic functions. The hormones may originate from extrahypothalamic sources such as the pancreas or adrenal gland.


Cerebral paragonimiasis--report of five cases. Five cases of cerebral paragonimiasis presenting with hemianopsia, convulsion, and gait disturbance are discussed. The cases were all in the chronic stage. The intradermal paragonimiasis reaction, complement fixation, and Ouchterlony tests were not useful for diagnosis. Computed tomography demonstrated calcifications in all cases in sites consistent with the foci of symptoms. Surgical treatment in two cases failed to improve symptoms.


Intracranial germinoma and Down's syndrome--case report. A germinoma in the basal ganglia developed in a 9-year-old boy with Down's syndrome, presenting as left hemiparesis. An initial computed tomographic (CT) scan demonstrated no notable abnormalities, but serial CT scans followed the entire course of tumor growth. Subtotal removal and irradiation achieved tumor remission. This is the first case reported of intracranial germinoma associated with Down's syndrome.


Unusual cranial metastasis from hepatoma presenting as isolated unilateral hypoglossal nerve paresis--case report. A 45-year-old male presented with a rare right hypoglossal nerve paresis due to an unusual cranial metastatic tumor from hepatoma. Despite multi-drug chemotherapy and arterial embolization, he died about 1 year later. Such metastasis should be considered as the cause of isolated unilateral hypoglossal nerve paresis.


Cystic pineocytoma--case report.

Cystic pineocytoma--case report. Pineocytoma and pineoblastoma, originating from pineal parenchyma, are rare and usually solid. An unusual case of totally cystic pineocytoma in a 37-year-old female is reported. The tumor showed neuronal differentiation and had a good outcome. Prominent calcification associated with pineocytoma and pineoblastoma is an useful finding to differentiate these from benign pineal cysts.


Steady state analysis of hypothalamic GnRH mRNA levels in male Syrian hamsters: influences of photoperiod and androgen. An in situ hybridization assay, utilizing a free floating technique was used to estimate the steady state levels of hypothalamic luteinizing hormone-releasing hormone (GnRH) mRNA levels in the brains of male golden hamsters maintained in different photoperiods. In situ histochemistry was performed using a 32P-labelled 66-nucleotide long oligomer complementary to the sequence of the human GnRH mRNA coding region. The oligonucleotide hybridized specifically to mRNA encoding the GnRH precursor as suggested by the distribution of labelled neurons and as shown by an RNAse protection assay on septal and preoptic-hypothalamic mRNA from gonadally regressed hamsters. To test the hypothesis that short-day photoperiods reduce GnRH synthesis, intact male hamsters or castrated males bearing subcutaneously inserted testosterone implants were exposed to long-day (14 h light:10 h dark) or short-day (10 h light 14 h dark) photoperiods for 4 weeks. Exposure to short day lengths never caused a decrease in GnRH expressing neurons and actually was associated with an increase in the number of radiolabelled cells specifically in the diagonal band of Broca/medial septum in the gonadally intact group. The mean number of grains per labelled cell for the short day animals similarly was not reduced from that seen in long day animals. The results are consistent with previous studies on photoperiod and GnRH content in the same brain regions and support the notion that the suppression of the synthesis of GnRH does not accompany the low levels of LH secretion observed during the early stages of reproductive quiescence in this species.


Epitope analysis of senile plaque components in the hippocampus of patients with Parkinson's disease. We conducted an epitope analysis of senile plaque (SP) proteins on hippocampal SPs in patients with Parkinson's disease (PD), using a library of antibodies to proteins implicated in the genesis of hippocampal SPs in Alzheimer's disease (AD). The library included antibodies to the beta-amyloid protein (beta-AP), domains outside the beta-AP in beta-amyloid precursor proteins (beta-APPs), ubiquitin, diverse neuronal cytoskeletal proteins, and polypeptides located mainly in axon terminals. We obtained samples of hippocampus at autopsy from 14 PD patients, 10 of whom were demented. As in the AD hippocampus, the SPs detected by conventional stains in five of the 10 demented subjects contained the beta-AP and flanking domains in beta-APPs as well as epitopes in tau, neurofilament proteins, and synaptophysin. Further, with the exception of the beta-AP, epitopes in the other proteins were confined to the coronas of SPs, while clathrin light chain, microtubule-associated protein 5, and neural cell adhesion molecules were almost undetectable or absent in the neuropil occupied by SPs. The same group of antibodies rarely labeled SPs in the other five demented PD subjects or in the four nondemented PD subjects, and conventional stains for amyloid and neurofibrillary pathology revealed rare SPs in these cases. Hence, when conventional stains reveal lesions diagnostic of AD in PD patients, the molecular features of the hippocampal SPs in these patients are the same as those in SPs of the AD hippocampus.


[Comparison of crystalloids and hydroxyethyl starch during normovolemic hemodilution. A study of hemodynamics and saturation measurement]. Normovolemic hemodilution at two different hematocrit values was performed in ten patients undergoing major surgery to evaluate changes of DO2, VO2 and CI. A Hct of 38% (low hemodilution) was reached by plasma replacement with ringer lactate infusion. A further hemodilution, a Hct of 30% (high hemodilution), was obtained by hydroxyethyl starch plus ringer lactate (1:2 ratio) infusion. A significant VO2 increase (p less than 0.01) occurred when hydroxyethyl starch plus ringer lactate infusions were employed as compared to ringer lactate alone. No changes in DO2 and CI were observed, the increase in VO2 measured during colloid infusion could suggest a better tissue perfusion and metabolic activity.


[Endoscopic laser therapy in the palliative treatment of cancer of the esophagus]. Frequently when dysphagia first becomes manifest, esophageal carcinoma is already unresectable. Endoscopic laser therapy is a recently introduced method which can be used for the palliative treatment of esophageal cancer. By using the method the tumour is destroyed and the esophagus is recanalized, there is low incidence of complications, and the patient can be fed naturally thus improving quality of life. The method has been perfected over the past years and now offers excellent results.


T gamma/delta lymphocytes in renal transplant recipients. T gamma/delta lymphocytes are able to perform allospecific cytotoxicity and natural killer cytotoxicity in vitro. However, very little is known about their function in vivo. To investigate the possible involvement of T gamma/delta lymphocytes in the immune response to renal allografts, fine-needle aspiration biopsies and peripheral blood of 15 renal transplant recipients were studied during the first 4 weeks after transplantation. In addition peripheral blood of patients before transplantation, half a year and one year after transplantation was studied. No increase in the percentage of T gamma/delta lymphocytes in the fine-needle aspiration biopsies, including those taken during acute rejection episodes, was found. A significant decrease in the percentage of T gamma/delta lymphocytes was observed in peripheral blood after transplantation. We conclude that T gamma/delta lymphocytes seem to play no major role in the immune response to renal allografts.


Staining of semithin tissue sections embedded in HPMA, quetol 523 and MMA. Various tissues fixed in a mixture of formaldehyde and glutaraldehyde, and embedded in an improved 2-hydroxypropyl methacrylate mixture were employed for studying the fine structures of cells and tissues by light microscopy. The embedding mixture contained Quetol 523 and methyl methacrylate as a plasticizer without a cross-linker. The catalyst was QCU-1. The mixture had a low viscosity, was easy to handle and penetrated readily and completely into the specimen, producing a homogeneous block from which it was easy to cut sections of 1-2 microns in thickness. A wide variety of stains have been employed with such sections and those reported here are hematoxylin-eosin, Azan and PAS. There was excellent preservation of alkaline phosphatase activity. A method of poststaining immunoperoxidase labeling was also applied to the mouse pancreas and examples of staining with insulin are included.


Cloning and structural analysis of the human c-kit gene. The recent identification of the mouse White spotting and Steel loci as genes encoding the c-kit receptor and its ligand, respectively, has shed light on the importance of this ligand and receptor in embryogenesis, melanogenesis and hematopoiesis. In order to determine if the c-kit proto-oncogene is involved in human disease, we isolated seven overlapping lambda recombinants, using a fetal brain cDNA, and characterized the normal human gene (KIT). The longest mapped transcript is 5230 bp, is alternatively spliced and includes 21 exons that span more than 70 kb of DNA. From the exon-intron structure, we have localized an alternative splice site to the 3' end of exon 9. The overall c-kit gene structure closely resembles that found in the CSF-1R gene (c-fms). This similarity includes a large first intron, the same number of exons containing translated sequence and very similar exon-intron boundaries. Using pulsed-field gel electrophoresis, we have linked KIT to the platelet-derived growth factor receptor A gene, with both residing on a 700-kb BssHI fragment. These data will allow investigation into the control of KIT expression and the potential to identify mutations or altered expression of this gene in human disease.


Expression of the c-erbB-3 protein in normal human adult and fetal tissues. c-erbB-3 is a member of the type I family of growth factor receptors which includes the epidermal growth factor (EGF) receptor and c-erbB-2. Whereas for EGF receptor and c-erbB-2 the expression patterns in normal tissues are well documented, there is currently little information available about the sites of c-erbB-3 expression. In order to examine the normal tissue distribution of c-erbB-3, polyclonal antibodies were raised against eight synthetic peptides corresponding to distinct sites on the intracellular domain of c-erbB-3. Of these, three produced antibodies which reacted with a 160-kDa protein on immunoblots of human embryonal cells (293 cells) transfected with the cDNA encoding c-erbB-3, and two of the three antibodies immunoprecipitated a protein of similar size from the same cells. These antibodies were used for immunochemical staining of a wide variety of normal human adult and fetal tissues employing formalin-fixed, paraffin-embedded material. The c-erbB-3 protein was identified in cells of the gastrointestinal, reproductive, respiratory and urinary tracts as well as the skin, endocrine and nervous system in a distribution distinctly different from that observed for EGF receptor and c-erbB-2. The level of expression of the mRNA for c-erbB-3 was also examined in extracts of a selection of fetal tissues. In general the sites of mRNA and protein expression were concordant.


Clonal characteristics of acute lymphoblastic cells derived from BCR/ABL p190 transgenic mice. The clonal and immunophenotypic characteristics of blood leukemic cells from BCR/ABL p190 transgenic mice were investigated. All cell populations evaluated in vivo and in vitro had B-lymphocyte progenitor immunophenotypes. Immunoglobulin (JH) rearrangement patterns provided evidence for clonal diversification at different sites in vivo. Multiple clones were established in vitro from two of these mice (nos. 730 and 753). These cells expressed BCR/ABL p190 protein tyrosine kinase (PTK) and were highly malignant on transfer to secondary recipients. Cells independently cloned in vitro shared identical immunophenotypes and clonal IgH rearrangements, but these were distinct from those of the dominant clones in the mouse from which they were derived. Nevertheless, in vitro clones from mouse no. 753 had an abnormal karyotype (chromosome 14 trisomy) in common with the dominant clone in blood, providing evidence for a hierarchy or clonal selection in vivo and in vitro. Two sets of in vitro clones proliferated independently of exogenous growth factors and stroma and released autocrine interleukin 7 growth factor activity. These data provide evidence for rapid divergent clonal evolution and selection of B-cell progenitors initiated by BCR/ABL p190, followed by other, secondary genetic events mirroring similar changes in the equivalent, highly malignant human leukemia Philadelphia (Ph)-positive/B-precursor acute lymphoblastic leukemia (ALL).


Structural organization of the murine c-kit proto-oncogene. The murine Kit receptor gene on chromosome 5 has been found to be frequently involved in germline mutations and rearrangements, leading to a characteristic set of severe developmental defects, known as the W phenotype. Here we describe the structure of the murine c-kit gene, based on restriction analysis of genomic phage clones and sequence determination of exon-intron boundaries. The Kit-coding region is distributed over 21 exons, most of which have sizes that range between 100 and 200 base pairs. The 3' non-translated sequence and the 3' end of the coding region form a single large exon, which encompasses 2.3 kb and is flanked by polyadenylation signals. The entire region spans a genomic distance of at least 70 kb. Though the exonic demarcations of c-kit show remarkable similarity to those of the human c-fms gene (which encodes the highly related colony-stimulating factor 1 receptor), no correlation could be found between the sizes of introns that separate homologous exon pairs. The data suggest that evolutionary pressures were confined to the conservation of structures of coding exons, whereas flanking regions were subject to large changes, owing to insertions and deletions. Finally, the analysis of the Kit genomic structure reveals that the inherited mutations of the Kit gene that have been reported thus far occur at various dispersed positions within the gene. Hence, the entire gene appears to have as yet unknown features which cause it to be frequently subject to mutations in murine germline tissues.


Voltage-gated Ca entry in isolated bovine capillary endothelial cells: evidence of a new type of BAY K 8644-sensitive channel. Isolated bovine capillary endothelial cells have been examined for voltage-dependent Ca entry. All cells displayed a low threshold activity, with the main characteristics of a T-type transient current, when examined using whole-cell recording for activation and inactivation and cell-attached conditions or inside-out patches for the elementary conductance (8 pS). 25% of the cells displayed an additional sustained current in 5 mM CaCl2 above -40 mV, which was enhanced by application of BAY K 8644, but almost insensitive to superfusion with nicardipine. Two types of channels (2.8 and 21 pS, in 110 mM BaCl2) were shown to have a BAY K 8644 sensitivity. The large conductance channels were L-type channels. The smaller events were elicited at more hyperpolarized potentials (by some 30 mV). Their mean open time was 16 ms in control conditions. In presence of BAY K 8644, additional long open times were observed (up to 100 ms as compared to 7.8 ms for the time constants of the slow mode of the L-type channel). We refer to these channels as SB channels: of small conductance and sensitive to BAY K 8644. In the presence of nicardipine, SB channels are not noticeably modified, in contrast to the L-type openings which are abolished. Also, SB open times are close to control values when nicardipine is added after a BAY K 8644 application. We suggest that, at physiological concentrations of divalent ions, an SB-type activity is elicited above -40 mV which generates the low threshold sustained current.


Arginine-vasopressin induces mode-2 gating in L-type Ca2+ channels (smooth muscle cells of the urinary bladder of the guinea-pig). The effect of arginine-vasopressin (AVP, 0.1 microM) on elementary Ca2+ channel currents (L-type) was studied in cell-attached patches with 10 mM BaCl2 as the charge carrier. At a constant potential of -30 mV, bath applied AVP increased the channel openness (NPo) by a factor of 4.7 +/- 3.0 (mean +/- SD, n = 9), the effect resulted from an increase in the frequency of opening (factor 2.5 +/- 0.8) and from a longer mean open time. Under control, openings longer than 5 ms contributed only 4% of the total, however, with the application of AVP this contribution increased to 29%. Under control, the open times were distributed along a single exponential (tau o1 = 0.8 +/- 0.4 ms), a double exponential distribution was obtained during AVP (tau o1 = 0.8 +/- 0.5 ms, tau o2 = 7.5 +/- 0.7 ms). The Ca2+ agonist BAYk8644 (1 microM) changed the open time distribution similarly to AVP (tau o1 = 1.0 +/- 0.5 ms, tau o2 = 9 +/- 2.8 ms). With 1 microM BAYk8644 in the bath, AVP did not significantly increase the relative contribution of long openings, however, AVP increased the frequency of openings by a factor of 2.0 +/- 1 (n = 6). The results are compatible with the idea that AVP can change the gating of L-type Ca2+ channels from mode 1 to mode 2.


Artiodactyl retroposons: association with microsatellites and use in SINEmorph detection by PCR. During a search of polymorphic microsatellites for bovine genome mapping, we found that microsatellites often occur as tails of artiodactyl C-A retroposon elements. In this element, C (85bp) is a tRNA derivative, while A (117bp) is of unknown origin. The A element also occurs as dimer element with a connecting 27bp linker sequence comprising hexanucleotide CACTTT repeats. In 10 clones (45% of those selected deliberately for dinucleotide repeats), the microsatellite motif is associated with the C-A retroposon. In 50% of 44 database artiodactyl C-A sequences, the element also has a microsatellite tail. The microsatellite is usually a simple (CA)n repeat, but in some cases it is an apparent derivative of the linker sequence CACTTT. All but one of 33 database dimer elements have trinucleotide repeat tails (AGC)n, n = 1-9. Microsatellites, retroposons, and their truncated versions (C and/or A) often occur as clusters. We derived the consensus sequence (202bp) of the C-A element, and designed four primers for inter-SINE amplification with the aim of finding SINEmorph polymorphisms. The method is potentially powerful for rapidly producing polymorphic markers for artiodactyl genome mapping.


Molecular cloning of two C/EBP-related proteins that bind to the promoter and the enhancer of the alpha 1-fetoprotein gene. Further analysis of C/EBP beta and C/EBP gamma. In an attempt to identify proteins that may regulate alpha 1-fetoprotein (AFP) gene expression, we screened a cDNA expression library from neonatal rat liver with two essential cis-elements of the AFP promoter and enhancer. We isolated two cDNAs which were found to correspond to leucine zipper proteins of the CC-AAT/enhancer binding protein (C/EBP) family: C/EBP beta and C/EBP gamma. The three related proteins C/EBP alpha, beta and gamma bind with indistinguishable specificity to multiple DNA sites in the promoter and the enhancer of the AFP gene. In addition, C/EBP beta and C/EBP gamma readily heterodimerize with each other as well as with C/EBP alpha. The mRNAs coding for C/EBP beta and C/EBP gamma are expressed in a wider variety of rat tissues than C/EBP alpha mRNA, including yolk sac and fetal liver. The steady-state levels of C/EBP alpha, beta and gamma mRNAs increase during liver development, in parallel with their respective gene transcriptional rates. The high levels of C/EBP beta and gamma mRNAs in rat yolk sac and fetal liver, where C/EBP alpha is poorly expressed, suggest that C/EBP beta and/or gamma could be preponderant or early regulators of the AFP gene in these tissues.


Novel mutants of 23S RNA: characterization of functional properties. Single point mutations corresponding to the positions G2505 and G2583 have been constructed in the gene encoding E.coli 23S rRNA. These mutations were linked to the second mutation A1067 to T, known to confer resistance to thiostrepton (1). Mutant ribosomes were analyzed in vitro for their ability to direct poly(U) dependent translation, their missence error frequency and in addition their sensitivity to peptidyltransferase inhibitors. It was evident that the mutated ribosomes had an altered dependence on [Mg2+] and an increased sensitivity to chloramphenicol during poly(U) directed poly(Phe) synthesis. In a transpeptidation assay mutated ribosomes were as sensitive to chloramphenicol as wild-type ribosomes. However, the mutant ribosomes exhibited an increased sensitivity to lincomycin. An increase in translational accuracy was attributed to the mutations at the position 2583: accuracy increased in the order G less than A less than U less than C.


Expression of the human cystic fibrosis transmembrane conductance regulator gene in the mouse lung after in vivo intratracheal plasmid-mediated gene transfer. As an approach to gene therapy for the respiratory manifestations of cystic fibrosis (CF), in vivo plasmid-mediated direct transfer of the normal CF transmembrane conductance regulator (CFTR) gene to the airway epithelium was investigated in mice. To evaluate the feasibility of this strategy, pRSVL, a plasmid composed of a firefly luciferase gene driven by the Rous sarcoma virus long terminal repeat (RSV-LTR), along with cationic liposomes was instilled into the trachea of C57BI/6NCR mice. With administration of 200-400 micrograms plasmid DNA, luciferase expression could be detected in the mouse lung homogenates for at least 4 wk. With this background, a CFTR expression plasmid vector (pRSVCFTR) constructed by replacing the luciferase cDNA from pRSVL with the normal human CFTR cDNA was evaluated in vivo in mice. Intratracheal instillation of pRSVCFTR with cationic liposomes followed by analysis of mouse lung RNA by polymerase chain reaction amplification (after conversion of mRNA to cDNA) using a RSV-LTR specific sense primer and a human CFTR-specific antisense primer demonstrated human CFTR mRNA transcripts from one day to 4 wk after instillation. Further, in vivo evaluation of beta-galactosidase activity after intratracheal administration of an E. coli lacZ gene expression plasmid vector directed by the cytomegalovirus promoter (pCMV beta) demonstrated that the airway epithelium was the major target of transfer and expression of the exogenous gene. These observations demonstrate successful plasmid-mediated gene transfer to the airway epithelium in vivo. This strategy may be feasible as a form of gene therapy to prevent the pulmonary manifestations of CF.


Isolation, purification and biochemical characterization of human placental interferons by tandem high-performance affinity chromatography. Human placental trophoblasts, fibroblasts and the trophoblast-derived malignant cell JAR are potent producers of interferons (IFNs) when stimulated with Sendai virus. The three cell lines produced different levels and compositions of IFN-alpha subtypes and IFN-beta. Anti-IFN globulins, Cibacron Blue F3GA and Concanavalin A were covalently immobilized on pressure-stable, macroporous polymeric matrices derivatized with vinyl sulphone (HEMA-BIO 1000 VS and HEMA 1000 VS). These supports were packed in biocompatible PEEK columns and were coupled with switching valves, to develop a tandem high-performance affinity chromatographic (HPAC) method for the isolation, purification and biochemical characterization of the IFNs produced in Sendai virus-stimulated human placental trophoblasts, fibroblasts and trophoblast-derived malignant cell, JAR, cultures. Silver-stained SDS-PAGE and gel densitometric analysis revealed the purity of the purified proteins to be between 94 and 98%. Specific activities of the purified IFNs ranged between 0.37-2.76 x 10(8) IU/mg of protein with cumulative recoveries between 90 and 92.2%. The purified IFN components exhibited quantitatively different antiviral activities in human and bovine cell lines. The utility of the tandem method for the purification and characterization of human type 1 IFNs produced from other cell lines are also discussed.


Detection of local structures in reduced unfolded bovine pancreatic trypsin inhibitor. The structure of BPTI and reduced BPTI in concentrated guanidinium HCl (GUHCl) in the presence of glycerol has been probed by measurements of dynamic nonradiative excitation energy transfer between probes attached to its amino groups. Interprobe distance distributions were obtained from analysis of donor fluorescence decay curves and used to characterize local structures in unordered states of the protein. Site specifically fluorescently labeled BPTI derivatives (1-n)BPTI (n = 15, 20, 41, 46) were used, each carrying a 2-methoxy-naphthyl-1-methylenyl group (MNA) at the N-terminal amino group of arg1 and 7-(dimethylamino)-coumarin-4-yl-acetyl residue (DA-coum) at one of its epsilon-NH2 groups of the lysine side chains. Analysis of donor fluorescence decay kinetics gave the interprobe distance distributions in the native and denatured states. The N-terminal-segment, residues 1-15, is in an extended conformation (with an average interprobe distance of 34 +/- 2 A) in the native state. Upon unfolding by reduction with DTT or beta-mercapto ethanol in 6 M GUHCl/glycerol mixture, the conformation of this segment relaxed to a state characterized by a reduced average interprobe distance and a larger width of the distances distribution. The average distance between residues 1 and 26, i.e., between the N-terminus and the turn of the twisted beta sheet element (residues 18-35), increased upon unfolding. At -30 degrees C in the above solvent, the distribution between these two sites was probably composed of two conformational subpopulations. About 45 +/- 20% of the molecules were characterized by a short interprobe distance (like the native state) representing a compact conformation, and 55 +/- 20% of the molecules showed large interprobe distances representing an expanded (unfolded) conformation. Thus local structures seem to exist in reduced denatured BPTI even under denaturing conditions in 6 M GUHCl/glycerol mixtures. Some of those structures are unstable in guanidinium isothiocyanate (GUSCN). The method introduced here is suitable for probing local structures and very long range interactions in unfolded proteins and for search for folding initiation sites (FISs) and early folding intermediates.


Mutating the charged residues in the binding pocket of cellular retinoic acid-binding protein simultaneously reduces its binding affinity to retinoic acid and increases its thermostability. Three-dimensional modeling of the complex between retinoic acid-binding protein (CRABP) and retinoic acid suggests that binding of the ligand is mediated by interaction between the carboxyl group of retinoic acid and two charged amino acids (Arg-111 and Arg-131) whose side chains project into the barrel of the protein. To assess the contribution of these amino acids to protein-ligand interaction, amino acid substitutions were made by oligonucleotide-directed, site-specific mutagenesis. The wild-type and mutant proteins were expressed in E. coli and subsequently purified. Like wild-type CRABP, the mutant proteins are composed mainly of beta-strands as determined by circular dichroism in the presence and absence of ligand, and thus presumably are folded into the same compact barrel structure as the wild-type protein. Mutants in which Arg-111 and Arg-131 are replaced by glutamine bind retinoic acid with significantly lower affinity than the wild-type protein, arguing that these two residues indeed interact with the ligand. The mutant proteins are more resistant to thermal denaturation than wild-type CRABP in the absence of retinoic acid, but they are not as thermostable as the CRABP-retinoic acid complex. These data suggest a model for CRABP-retinoic acid interaction in which the repulsive forces between the positively-charged arginine residues provide conformational flexibility to the native protein for retinoic acid to enter the binding pocket. Elimination of the positively-charged pair of amino acids produces a protein that is more thermostable than wild-type CRABP but less effective at ligand-binding.


Effects of nimodipine on biogenic amines in discrete brain areas. Discrete brain sections were obtained from rats after once or repeatedly given intraperitoneal nimodipine at doses ranging from 2.5 to 40 mg/kg. The single or the last treatment was carried out at 2-8 h before killing. The biogenic amine and metabolite content of the tissue samples were determined by high-performance liquid chromatography with electrochemical detection. The nimodipine-induced effects, chiefly regarding the brainstem, the thalamus-midbrain and the striatum, consisted of both an increase in 5-hydroxyindoleacetic acid (5-HIAA) levels and a decrease in dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels. Since the 5-HIAA/5-hydroxytryptamine (5-HT) ratio was increased, whereas the DOPAC/dopamine (DA) and HVA/DA ratios were decreased, it was argued that nimodipine activated the serotonergic and inhibited the dopaminergic systems. The first effect was enhanced by fasting and, likewise, by drug administration repeated for 5 days, while the second was not affected by fasting, but disappeared after 5 days' treatment. The data obtained appeared to substantiate the usefulness of nimodipine to treat some disorders of the central nervous system.


Release of calcitonin gene-related peptide and tachykinins from the rat trachea. The release of calcitonin gene-related peptide (CGRP), neurokinin A (NKA) and substance P (SP) from intralumenally perfused rat trachea was examined in vitro. In accord with the relative tissue levels of the respective peptides, capsaicin (10(-8) to 10(-5) M) and K+ (120 mM) added to the perfusate resulted in a concentration-dependent increase in the levels of CGRP and NKA, and to a minor extent SP, in the perfusates. Sequential exposure of the trachea to capsaicin revealed a concentration-dependent tachyphylaxis of CGRP release. Thus, 40 min after the application with capsaicin 10(-5) M, a second exposure to capsaicin at the same concentration, or K+ 120 mM, did not evoke CGRP release. In contrast, prior stimulation with K+ 120 mM significantly enhanced the CGRP release induced by a second stimulation with K+ 120 mM or capsaicin 10(-5) M. Capsaicin- and K(+)-induced peptide release was diminished or abolished in the absence of Ca2+. HPLC analysis of CGRP in release materials revealed that there was a single peak which eluted in the same fraction as synthetic rat CGRP. These data demonstrate that CGRP, NKA and SP exist in releasable, capsaicin-sensitive pools in terminals which lie within the proximal lumen of the trachea.


Substance P inactivation by transglutaminase in vitro. Gamma(glutamyl5)spermine derivative of substance P (Spm-SP) was synthesized in vitro in the presence of purified guinea pig liver transglutaminase and Ca2+. The spermine adduct of the neuropeptide was purified by HPLC on a reversed-phase column and characterized by fast atom bombardment mass spectrometry. The biological activities of Spm-SP were tested by assaying, in comparison with substance P, its ability to induce both the contractions of smooth muscle in vitro and the edema formation in vivo. Spm-SP was shown not to elicit contractile responses in the isolated rat stomach strip and duodenum and not to antagonize the spasmogenic effect evoked by the native neuropeptide. Furthermore, Spm-SP was unable, when administered into rats by plantar injection, either to provoke an acute inflammatory response in the hind limb or to antagonize the edema formation induced by a concurrent administration of substance P. These results indicate that the introduction of a large size hydrophilic moiety at the glutamine5 level negatively affects the ability of the neuropeptide to bind to its receptor(s), thus supporting the view that the hydrophobic middle portion of substance P plays a key role in receptor recognition.


Ataxia telangiectasia in the British Isles: the clinical and laboratory features of 70 affected individuals. Seventy individuals with ataxia telangiectasia were studied: 29 females and 41 males with an age range of 2 to 42 years. The majority (43/68) presented by 3 years of age with truncal ataxia. All had progressive, handicapping neurological symptoms exhibiting ataxia (70/70), ocular motor apraxia (70/70), an impassive face (70/70), dysarthria (70/70), chorea (68/70), dystonia (55/70) and peripheral neuropathy (50/70). Clinical immune deficiency was present in 43 of 70 patients. Ocular telangiectasia were seen in all but one case and excessive thinness in 54 of 70. The mean age of loss of walking was 10 years and of writing 8 years. All 60 tested showed increased sensitivity to ionizing irradiation, 43 of 48 had an elevated alpha-fetoprotein level and 14 of 21 had an immunoglobulin deficiency. Although there was a marked variation in disease findings sibs were always similar. The heterogeneity seen seems at odds with the unilocus linkage of ataxia telangiectasia to 11q23.


Factors predictive of completion of treatment and survival after palliative radiation therapy. Ninety-seven patients who underwent radiation treatment for metastatic carcinomas and sarcomas were evaluated to define prognostic factors that may reliably help determine survival and probability of completing a course of palliative radiation therapy. Actuarial and logistic models were used for analysis. Predictor variables included age; sex; symptoms; primary site of disease; pathologic diagnosis; prior metastatic disease, treatment, and response; solitary versus multiple metastatic sites; location of metastasis; status of primary lesion; interval between initial diagnosis and treatment for metastatic disease; and Karnofsky performance score (KPS). The interval between primary diagnosis and metastatic treatment, and KPS, were significant variables for 22 patients (23%) who failed to complete their planned radiation treatment. KPS was consistently significant for probability of survival at 2-, 4-, 8-, and 16-month intervals. At 8 months and 16 months, site of primary disease was significant, and at 16 months, solitary site of metastasis was also significant. Conventional factors, especially KPS, are useful in predicting the likelihood of completing radiation therapy and of subsequent survival for patients undergoing palliative treatment.


Tissue distribution and innervation pattern of peptide immunoreactivities in the rat pancreas. The distribution of calcitonin gene-related peptide (CGRP), substance P/tachykinin (SP/TK), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and gastrin-releasing peptide (GRP) immunreactivities (IR) in the rat pancreas was investigated using radioimmunoassay and immunohistochemistry. CGRP, NPY and VIP tissue contents are much higher than GRP and SP/TK concentrations. Peptide-containing nerves are distributed to both the exocrine and endocrine pancreas. However, differences exist in terms of density and targets of innervation for each peptidergic system. In the acini and through the stroma, fibers IR for CGRP, NPY and VIP are greater than GRP- and SP/TK-containing processes. The vasculature is supplied by a prominent NPY, CGRP and, to a lesser extent, SP/TK innervation. VIP-IR is found occasionally, and GRP-IR is never detected, in fibers associated with blood vessels. Around ducts, CGRP- and NPY-positive neurites are greater than SP/TK- greater than or equal to VIP-IR fibers, whereas GRP-containing nerves are not visualized. In the islets, the density of peptidergic nerves is: VIP-, GRP- greater than or equal to CGRP-IR greater than NPY or SP/TK. In intrapancreatic ganglia. VIP- and, to a lesser extent, NPY-IRs are found in numerous neuronal cell bodies and in nerve fibers; GRP-IR is present in numerous nerve processes and in few cell bodies; CGRP- and SP/TK-IRs are detected only in fibers wrapping around unlabeled ganglion cells. The majority of CGRP-IR fibers contain SP/TK-IR. The existence of differential patterns of peptidergic nerves suggests that peptides exert their effects on pancreatic functions via different pathways.


Changes in serum and salivary amylase during radiotherapy for head and neck cancer: a comparison of conventionally fractionated radiotherapy with CHART. The changes in serum amylase that occur when radiotherapy is given in the treatment of head and neck cancer has been studied in 41 patients, 29 treated by CHART and 12 by conventionally fractionated radiotherapy. The peak rise in serum amylase following the start of treatment is seen earlier and is greater in the patients receiving continuous hyperfractionated accelerated radiotherapy (CHART). The serum amylase returns to normal earlier in the CHART patients so that the area under the curve is the same for both groups. The difference probably reflects the more rapid delivery of treatment to the patients receiving CHART. A close correlation between the peak rise in serum amylase and the amount of parotid tissue in the treatment volume is demonstrated. For six patients the total amount of amylase secreted by the parotid gland during CHART was measured and found to decline rapidly within a few days of the start of radiotherapy. The rise in serum amylase that results from the irradiation of salivary tissue provides a unique biochemical measure of an early radiation effect in a normal tissue. This probably reflects the interphase cell death of serous salivary cells. Although these immediate changes are of considerable interest they may not relate to the late effects of radiation on salivary gland function.


The significance of age of onset for schizophrenia. Age of onset is the single most important characteristic of schizophrenia that could yield clues to its origin. To identify the age of onset, however, the onset of the pathological process must be determined. This process may have more than one component occurring at distinctly different times in the life of an individual. Nevertheless, many studies, using either the first appearance of psychosis or the age of first hospitalization, have found gender, familial, and other "age of onset" differences among patients with schizophrenia. These differences may aid in examining genetic mechanisms for schizophrenia.


A neurodevelopmental approach to the classification of schizophrenia. The conventional distinction between schizophrenia and manic depression has received little objective support from recent studies of phenomenology, outcome, or familial homotypy. Instead, much clinical, epidemiological, and morphological evidence suggests that within the broad range of Schneiderian schizophrenia there exists one form (congenital schizophrenia) that can be distinguished from other types, the manifestations of which are confined to adult life. We hypothesize that congenital schizophrenia is a consequence of aberrant brain development during fetal and neonatal life. Such patients show structural brain changes and cognitive impairment, and in their male predominance, early onset, and poor outcome, they reflect Kraepelin's original description of dementia praecox. We contend that adult-onset schizophrenia is itself heterogeneous. One important component is a relapsing and remitting disorder that is more frequent in females than in males, exhibits positive but not negative symptoms, and has much in common etiologically with affective psychosis. There also exists a very-late-onset group in which degenerative brain disorder is implicated.


[Cation channels formed in lipid bilayer by Pinellia ternata lectin]. Pinellia ternata lectin (PTL), a protein exhibiting hemagglutination activity and carbohydrate binding specificity to mannan was purified from rhizome of Pinellia ternata. In this work the actions of PTL on artificial lipid bilayer were investigated by means of the two-compartment system of Mueller and Rudin. The lipid bilayer with resistance more than 10G omega was formed by a solution of lecithin and cholesterol (20 mg/ml and 5 mg/ml respectively) in N-decane. The electrical properties of the lipid bilayer were investigated in voltage clamp mode. Several minutes after the addition of PTL (2 micrograms/ml) in one compartment the channel-like noise as well as a decrease of the resistance of the bilayer were observed. These actions were inhibited by mannan significantly. The resistance increase of the bilayer with PTL-channels could be observed from 2G omega to control level (greater than or equal to 10 G omega) immediately after addition of 40 micrograms/ml mannan. The discrete conduction steps were recorded at low concentration of PTL and at low holding potential. The predominant unit conductance was 35pS in symmetric KCl solution of 100 mmol/L. The selectivity of PTL-channel was estimated from Goldman-Hodgkin-Katz equation by measurement of the reversal potential in an asymmetrical salt solution. The results showed that PTL-channels were cation selective.


[On the relation between substance P and physostigmine on cardiovascular control in rabbit brain]. Using an electromagnetic flowmeter technique, the cardiac output as measured by blood flow in the aortic arch was measured during intracerebroventricular injection (icv.) of Substance P (SP) and physostigmine in 47 anesthetized rabbits. Carotid artery blood pressure and heart rate were recorded. After icv of SP (20 micrograms/20 microliters) or physostigmine (60 micrograms/20 microliters), both cardiac output and mean artery pressure were increased. Pretreatment with icv. of atropine (150 micrograms/20 microliters) did not alter the effect of SP, but the effect of physostigmine was blocked by pretreatment with SP blocker (25 micrograms/20 microliters). These findings suggest that cerebral SP is involved in cholinergic mechanisms on the central control of blood pressure.


Enteral nutrition with supplemental arginine, RNA, and omega-3 fatty acids in patients after operation: immunologic, metabolic, and clinical outcome. The individual nutrients arginine, RNA, and omega-3 fatty acids improve immune function, but prospective trials have not demonstrated their effects on clinical outcome. Patients (n = 85) who underwent operation for upper gastrointestinal malignancies were randomized to receive the supplemental diet or a standard enteral diet after surgery. Clinical patient characteristics were similar between the two groups. Mean caloric intakes (1421 vs 1285 kcal/day) were similar between groups. Mean nitrogen intakes (15.6 vs 9.0 gm/day) and nitrogen balances (-2.2 vs -6.6 gm/day) measured in the first 20 patients were significantly greater in the supplemented group than in the standard group (p = 0.05). In vitro lymphocyte mitogenesis was measured in the first 31 patients and was decreased on postoperative day 1 in both groups, but normal levels were regained only in the supplemented group. In the cohort of 77 eligible patients, infectious and wound complications occurred significantly less often (11% vs 37%) in the supplemented group than in the standard group (p = 0.02). Linear logistic models for infectious/wound complications with control for the amount of nitrogen suggested (p = 0.10) dietary treatment as the major factor. Mean length of stay in the hospital was significantly shorter (p = 0.01) for the supplemented group (15.8 +/- 5.1 days) than for the standard group (20.2 +/- 9.4 days). These results suggest that postoperative enteral nutrition with supplemental arginine, RNA, and omega-3 fatty acids instead of a standard enteral diet significantly improved immunologic, metabolic, and clinical outcomes in patients with upper gastrointestinal malignancies who were undergoing major elective surgery.


[Hepatitis serology: use and interpretation]. Presently five viruses causing hepatitis are known, the hepatitis viruses A (HAV), B (HBV), C (HCV), D (HDV) and E (HEV). The genomic structure is known of most of all these viruses as well as some of their structural and regulatory gene products. Using radio- and enzyme immunoassays viral antigens can be detected for HBV and HDV as well as specific antibodies against all the five viruses. The results of these tests are the basis for the diagnosis and the follow-up of these infections but differ in their accuracy for each given virus. Concerning HIV one can differentiate between an ongoing or recently passed infection and immunity. Concerning HBV (and HDV), an ongoing infection at various stages can be distinguished from immunity. Such distinctions are not possible with respect to HCV except when also applying the expensive and cumbersome method of the polymerase chain reaction. In this paper the most important characteristics of the hepatitis viruses are given and the behaviour of the various viral markers during the infections and their consequences are described.


[Interferon treatment of chronic viral hepatitis]. The interferon treatment of chronic viral hepatitis leads to a measurable inhibition of replication of the various hepatitis viruses. In adult patients with chronic HBe-Ag-positive hepatitis B, the seroconversion rate is tripled by therapy when compared with untreated controls. In chronic hepatitis C, biochemical remission is achieved in about 50% of patients, but there is a high tendency to relapse. Interferon treatment has little effect in hepatitis B virus infection transmitted perinatally and in chronic delta hepatitis. Better knowledge of prognostic variables and deeper insights into the mechanisms of chronic hepatitis C virus infection should help both to increase rates and duration of remissions and to reduce the rates of relapse in the future.


[Palliative treatment of liver tumors].

[Palliative treatment of liver tumors]. Metastases of extrahepatic primaries are the most prevalent cause of malignant cells found in the liver. This overview is focusing on different regional and systemic palliative treatment modalities according to the histology of the primary tumor with special emphasis on metastastic colonic cancer. Furthermore indications and contraindications of the various treatment options are discussed.


Cutaneous ureterostomy in adults.

Cutaneous ureterostomy in adults. In the patient with metastatic carcinoma, urinary diversion is usually achieved with indwelling ureteral stents or placement of a percutaneous nephrostomy tube. Most forms of surgical diversion carry an unacceptable morbidity rate, especially in the debilitated patient. Over a fifteen-year period (1974-1989), 29 adult patients with pelvic malignancy (32 ureters) underwent palliative cutaneous ureterostomy. This previously reported technique involves transverse nephropexy and construction of a stoma using a small skin flap. Indications included ureteral obstruction or severe urinary tract symptoms. Hydroureter, often considered a precondition for this procedure, was not present in several patients and was not a prerequisite to success. Complications related to the procedure included one postoperative death due to stroke, one death due to uremia and sepsis, and one instance of severe renal arterial stenosis resulting in renal failure. Preservation of renal function was possible in the 10 patients known to have survived from one to thirteen years postoperatively; only 3 patients eventually required stomal revision. By adherence to the surgical techniques described, the usually high incidence of stomal stenosis was avoided. Our experience reveals that although the indications for cutaneous ureterostomy are limited, this procedure can provide an alternative to permanent nephrostomy drainage or to a higher risk intestinal urinary diversion in carefully selected patients with a reasonable life expectancy.


Guided brachytherapy for treatment of confined prostate cancer. A total of 133 patients underwent transperineal ultrasound-guided iodine 125 seed implantation for Stages A and B prostate cancer with a twenty-seven-month follow-up. There has been no mortality and our morbidity is no more than experienced after transurethral resection of the prostate. By using a Mick applicator our operating time is well under one hour, and our patients go home the same day without a Foley catheter. Our results indicate that patients with PSA values of less than 20 ng/mL (Yang method) and/or Gleason scores of 6 or less are excellent candidates for brachytherapy. By subdividing the percentage of normal PSA values in the follow-up periods according to the patient's original PSA value, further credence is given to the PSA value as a strong aid in staging when the Gleason score is 6 or less. Although the follow-up at twenty-seven months is small, our preliminary results indicate that brachytherapy is a viable option to radical surgery in those patients who are not good candidates for surgery or who prefer nonsurgical treatment.


Pulmonary embolism from left subclavian vein thrombus following suprapubic prostatectomy. Deep venous thrombosis (DVT) of the axillary and subclavian veins accounts for approximately 1-2 percent of all recorded deep venous thrombosis. Pulmonary embolism from an upper extremity DVT has been reported to vary between 2 percent and 35.7 percent. We report the occurrence of a left subclavian vein DVT with subsequent nonfatal pulmonary embolism in a sixty-two-year-old patient twenty-four hours following suprapubic prostatectomy. A review of the literature is presented, along with pathophysiology, diagnosis, and treatment.


Response to second-line hormonal manipulation monitored by serum PSA in stage D2 prostate carcinoma. Changes in prostate-specific antigen (PSA) have been demonstrated to accurately assess response to initial hormone deprivation in metastatic prostate cancer patients. The role of PSA in monitoring response to second-line hormonal treatment has not been documented. In a group of 20 patients with an initial response to androgen deprivation and subsequent relapse, we monitored PSA levels before and after second-line therapy. Ten patients had a clinical response. Four had a more than 90 percent decrease in serum PSA compared with the level at initial progression. This clinical response was maintained for a mean of eighteen months. Six patients had a PSA decrease less than 90 percent; their clinical response was of a mean 5.5 months. Ten patients had no change or increase in PSA. Seven had no clinical response, and 3 responded for an average of four months. Although production of PSA might be under endocrine control, changes in PSA are useful for monitoring response to second-line hormonal therapy.


Intrathecal administration of substance P in the rat: the effect on bladder and urethral sphincteric activity. At the lumbosacral spinal cord level in the rat, substance P-positive neurons are present in dense concentration in the dorsal horn and the sacral parasympathetic nucleus. We undertook the present study to investigate the effect of intrathecal substance P (10 micrograms at the L6-S1 level) on urinary bladder and urethral sphincteric activity and to compare these effects with those of intravenous and intra-arterial administration. Three different bladder pressure responses were triggered by intrathecal substance P: (A) an immediate, strong bladder contraction (n = 5); (B) augmentation of the micturition reflex, as indicated by strong detrusor contractions in response to intravesical saline perfusion (n = 4); and (C) a slow, gradual increase to a high, steady peak (n = 8). The sphincteric electromyographic (EMG) activity was consistently increased. When substance P was given intravenously (n = 10) and intra-arterially (n = 3), the form, duration, and maximal amplitude of bladder contractions (owing to a direct smooth-muscle action) were comparable with those of Group A. The effects in intrathecal Groups B and C suggest that substance P provides a tonic influence on motor horn cells and on the preganglionic neurons in the sacral parasympathetic nucleus at the lumbosacral spinal cord level, where neuronal circuits controlling bladder and sphincteric activity are located.


[Combined surgical-interventional procedure in congenital heart defects with postoperative, left ventricular dysfunction]. Creation of a communication between the left and right atrium to decompress the left ventricle can be life-saving after corrective surgery in some patients with congenital heart disease and small left ventricle. After adaptation of the left ventricle, surgical closure of this anastomosis becomes mandatory. We report four patients (2.9 to 8.2 kg) where non-surgical, transcatheter-closure was performed in the first year of life using the Rashkind-PDA-Occluder System (USCI). In a newborn with d-transposition of the great arteries (2.9 kg), in a patient with atrial septal defect (5.1 kg), and in a patient with tetralogy of Fallot with atrial septal defect (4.9 kg) atrial septal defects were not completely closed during correction because of left-ventricular dysfunction. Five to 12 days postoperatively transcatheter closure was performed utilizing the 8F-Rashkind-PDA-Occluder System in the first two patients (2.9 kg and 4.9 kg) and by use of the 11F-Rashkind-PDA-Occluder System in the third patient (4.9 kg). In the patient with supracardiac total anomalous pulmonary venous drainage via a vertical vein, the vertical vein was not closed during surgical correction at the age of 2 months. Transcatheter closure was performed 6 months later using the 11F-Rashkind-PDA-Occluder-System. It is concluded that small, defined atrial septal defects can be closed, even in newborns, using the Rashkind-PDA-Occluder System (USCI) and, furthermore, that large vessels, even without luminal narrowing can be occluded completely. Thus, this combined "surgical-interventional approach" is not limited by age and could reduce the risk of surgical correction in patients with congenital heart disease and small left ventricle.


Effect of adjuvant formulations on the selection of B-cell epitopes expressed by a malaria peptide vaccine. Because subunit vaccines may create artificial epitopes, the goal of this study was to concentrate the immune response toward protective epitopes contained in a peptide, [(NAGG)5]Y. This peptide consisted of five repeat sequences of the immunodominant epitope of the circumsporozoite protein of the simian malaria parasite Plasmodium cynomolgi and a terminal tyrosine. It was conjugated to bovine serum albumin and mixed with various adjuvant formulations, including block copolymers L121, L141, L180.5 and a lipopolysaccharide (LPS). Outbred mice were vaccinated with seven vaccine formulations. Postvaccination sera from each group of mice were then pooled, and antibody responses against peptide [(NAGG)5]Y or (NAGG)5 were tested in an enzyme-linked immunosorbent assay and against sporozoites of P. cynomolgi in an indirect fluorescent antibody assay. Although all groups elicited a high antibody response to the peptide [(NAGG)5Y], the presence of the tyrosine induced a different antibody response to the peptide (NAGG)5, and formulations containing LPS alone did not induce an antibody response to either (NAGG)5 or P. cynomolgi sporozoites. The formulation including both LPS and the copolymer L121 was the only one to inhibit the development of P. cynomolgi sporozoites in rhesus monkey hepatocytes by 50%. These results suggest that vaccine formulations influence B-cell epitope selection.


[Neo-angiogenesis after transplantation of a free skeletal muscle flap to the myocardium of the dog]. For the study of the process of neovascularization the effects of the transplantation of a free skeletal muscle flap on the heart of the dog were studied. For this purpose, in 14 mongrel dogs a myocardial infarction of the anterior wall of the heart was produced by the selective injection of microspheres into the left anterior descending artery. In 12 of those, on the area of infarction 4 weeks later a free pectoralis muscle flap was transplanted with its pars anterior in contact with the myocardium (group A). The arterial anastomosis was achieved with the internal mammary artery, the venous flow directed into the right atrium. In 2 other dogs (group B) the pectoralis flap was transplanted with irt pars anterior on a healthy myocardium. In 2 further dogs (group C) the pectoralis flap was transplanted on a infarcted myocardium with its pars posterior, i.e. the muscle fascia interponed. 16 weeks later a microcorrosion-cast-preparation and histological examinations showed in group A and B a prominent capillary network penetrating from the muscle into the myocardium, whereas in group C the muscle fascia inhibited this process. Thus, for the process of neovascularization the "bloody" contact between a skeletal muscle graft and the myocardium is necessary. Results are discussed with regard to possible factors inducing neoangiogenesis.


Insulin-like growth factor binding protein-3 in normal pubertal girls. IGFBP-3 concentrations rise in the second decade of life. To test the hypothesis that the stage of pubertal development, independent of chronological age, was associated with these increases we measured serum IGFBP-3 concentrations by radioimmunoassay in 324 sixth and seventh grade girls (12.3 +/- 0.7 years) at the beginning of a multisite school-based health curriculum. The mean (+/- SD) serum IGFBP-3 among the 242 girls with complete data was 4.0 +/- 0.7 mg/l. Pubertal stage was significantly associated with IGFBP-3 (p less than 0.0001, ANOVA). Mean concentrations rose from 3.5 +/- 0.7 mg/l among those with the earliest pubertal stages to 4.2 +/- 0.7 mg/l among the mature girls. IGF-I and IGFBP-3 concentrations were significantly correlated (Spearman's r = 0.43, p less than 0.0001). After controlling for the association between pubertal development and IGFBP-3 concentrations, only the waist/hip ratio, among the various measures of body composition, was significantly associated with IGFBP-3 concentration (Spearman's r = -0.23, p = 0.0002). Likewise, none of the measures of nutrition: intake of total calories, protein, fat and carbohydrate; serum iron; red cell mean corpuscular volume; or cholesterol; were significantly associated with IGFBP-3 concentrations. There was, however, a small, but significant association between IGFBP-3 concentrations and both serum transferrin and blood hemoglobin concentrations. Pubertal stage has a significant impact on IGFBP-3 concentrations and those attempting to utilize IGFBP-3 concentrations during adolescence should be cognizant of the subject's pubertal stage.


Triiodothyronine (T3) stimulates insulin-like growth factor (IGF)-1 and IGF binding protein (IGFBP)-2 production by rat osteoblasts in vitro. Osteoblast-like cells prepared from neonatal rat calvariae and grown under serum-free conditions produce IGF-1 and IGFBPs. In contrast to growth hormone, T3 and PTH increased both IGF-1 mRNA expression and net IGF-1 release in calvaria cells. In addition, they stimulated net production of IGFBP-3 and of an IGFBP with an apparent molecular weight of 32 kDa which was recognized by an antiserum against rat IGFBP-2. Bone cells expressed remarkably high levels of mRNA for IGFBP-2, the predominant IGFBP in serum of newborn rats. T3 at low physiological concentrations but not growth hormone stimulated IGFBP-2 mRNA expression and IGFBP-2 production in bone cells in vitro. Thus, IGFBPs are differentially regulated by these hormones and may play an autocrine/paracrine regulatory role in bone.


Amyloid beta protein precursors with kunitz-type inhibitor domains and acetylcholinesterase in cerebrospinal fluid from patients with dementia of the Alzheimer type. We used the ELISA to measure the concentration of amyloid protein precursor with Kunitz type trypsin inhibitor domains (APPI) in CSF of dementia of the Alzheimer type (DAT) and examined the correlation of APPI with acetylcholinesterase (AChE) and somatostatin (SRIF). We found the APPI concentration in CSF of DAT to be significantly elevated compared with that of multi-infarct dementia and controls. We could significantly correlate APPI with AChE, but not correlate APPI with SRIF. The present results suggest that measurement of CSF APPI levels may be useful for diagnosis of DAT and the change of APPI may closely be associated with abnormality of acetylcholine system in DAT that has been reported.


Immunocytochemical demonstration of interphotoreceptor retinoid-binding protein in cerebellar medulloblastoma. Previously, immunoreactive rod-opsin and S-antigen (arrestin), two highly characteristic markers of retinal photoreceptors and pinealocytes, were shown to be present in certain medulloblastoma cells. It, thus, has been suggested that such cells differentiate along the photoreceptor lineage. This is corroborated in the present immunocytochemical investigation using antibodies against another photoreceptor-cell marker, the interphotoreceptor retinoid-binding protein (IRBP). As shown in preparations of human retina and pineal organ, IRBP can be successfully demonstrated in formalin-fixed and paraffin-embedded tissue: the IRBP immunoreaction is located to the outer and inner segments of retinal photoreceptor cells and to perikarya of certain pinealocytes. Examination of formalin-fixed, paraffin-embedded biopsy specimens of 66 cerebellar medullo-blastomas revealed varying numbers of IRBP-immuno-reactive tumor cells in 19 cases that were formerly shown to contain rod-opsin and S-antigen immunoreaction. IRBP-immunoreactive tumor cells were also found in a retinoblastoma and a pineocytoma, but not in neuroblastoma, ganglioneuroblastoma, glioblastoma, oligodendroglioma and astrocytoma. The results indicate: (1) cerebellar medulloblastomas are heterogeneous in their differentiation potential; (2) one type of medulloblastoma displays photoreceptor characteristics; (3) this type appears to be closely related to retinoblastoma and pineal cell tumors; and (4) all three types of tumors may display additional common features to be explored in future studies.