Influence of exogenous gangliosides on the three-dimensional sprouting of goldfish retinal explants in vitro. To investigate the 3-dimensional outgrowth of ganglion cells of normal and regenerating goldfish retina, retinal explants were cultured in a serum free 3-D fibrin matrix. Daily applications of exogenous gangliosides (GM1), injected either intraocularly (i.o.) or intraperitoneally (i.p.) had no significant effect on the sprouting activity of retinal explants prepared from lesion-activated goldfish whose corresponding optic nerve had been transected. However, in normal, unlesioned animals, a local i.o. injection of GM1 or mixed gangliosides led to a significant enhancement of the basal retinal sprouting activity as compared to controls, which were injected with a 0.9% NaCl solution. This ganglioside related stimulation was maximal after i.o. injection of low concentrations (3 micrograms/eye), didn't occur at high concentrations (30 micrograms/eye) and was similar to the response obtained after i.o. injection of NGF or insulin. I.o. injected phospholipids had no or a slightly inhibitory effect on the sprouting activity as compared to NaCl controls. Daily in vivo i.o. injections of the monoclonal antibody Q211, specifically recognizing c-pathway polysialogangliosides, led to a dose dependent inhibition of the in vitro sprouting of goldfish retina explants. In summary, these data suggest an involvement of gangliosides in the complex process of induction of neuronal sprouting.


beta-Amyloid peptide in vitro toxicity: lot-to-lot variability. beta A4 peptide (beta AP) accumulates in amyloid plaques of Alzheimer's disease and may contribute to neuronal degeneration. Conflicting observations have been reported regarding the direct in vitro and in vivo neurotoxicity of beta AP. We have assessed in vitro beta AP toxicity in high density primary rat hippocampal cultures and found marked lot-to-lot differences in the neurotoxic properties of beta AP. One lot of beta AP from a commercial supplier resulted in significant direct neurotoxicity at 10 microM, while 2 other lots from the same supplier were essentially nontoxic. Three additional lots of beta AP from unrelated sources were also nontoxic at 10 microM. Initial biochemical characterization has not yet revealed any marked differences among the various lots of beta AP. Low levels of endotoxin (ca., 1 EU/ml) were detected in several beta AP preparations but did not correlate with neurotoxicity. Our observation that lot-to-lot variability of beta AP occurred even under identical in vitro culture conditions may account for part of the present controversy in this area.


Susceptibility of myelin proteins to a neutral endoproteinase: the degradation of myelin basic protein (MBP) and P2 protein by purified bovine brain multicatalytic proteinase complex (MPC). Multicatalytic proteinase complex (MPC) was isolated from bovine brain and the susceptibility of myelin basic protein (MBP) and P2 protein of bovine central and peripheral nervous system was examined. SDS-polyacrylamide electrophoretic analysis of purified MPC revealed protein bands of molecular weight ranging from 22-35 kDa. The enzyme is activated by SDS at a concentration less than 0.01%. Upon incubation with MPC, purified MBP and P2 proteins were degraded into smaller fragments. There was a 57% and 100% loss of MBP at 2 and 6 hours of incubation. The P2 protein which is not susceptible to any endogenous non-lysosomal enzyme thus far studied was digested into small peptide fragments only in the presence of SDS (0.01%) and not in its absence. These results indicate that MPC which is active at physiological conditions may have a role in the turnover of myelin proteins and in demyelinating diseases.


[Evaluation of NOR changes in patients with chronic active hepatitis treated with IFN and untreated]. The Authors consider 30 patients with ECA; 15 of these were treated with IFN and 15 were not treated. The Authors think that the statistically significant difference between patient groups, is a parameter of the pharmacological effects on the horx.


[Primary non-Hodgkin's lymphoma of paranasal sinuses. Review of the literature and a case report]. A case of primary extranodal non-Hodgkin's lymphoma arising in paranasal sinuses, in an 89 year-old woman is reported. Histologically it was lymphoplasmacytoid, pleomorphic, diffuse, at intermediate grade of malignancy according to "Working formulation for clinical usage" in I EB stage. Complete resolution was obtained with polychemotherapy according to the scheme ProMACE-CytaBOM. After a wide review of the literature, the Authors emphasize incidence, prevalence, clinical symptoms (especially local), the different beginning points, prognostic and therapeutic factors, and the most frequent associations with other illnesses. Agreeing with other Authors, the necessity of using aggressive polychemotherapy is stressed, also in elderly patients, even if at the beginning of the illness.


[Effects of long-term iloprost therapy on Raynaud's phenomenon in progressive systemic sclerosis]. One of the most appealing current pathogenetic concepts is that progressive systemic sclerosis (PSS) is a reaction to repeated episodes of endothelial cell injury. Injury of small arteries and capillary endothelium initiates reactions which involve increased permeability of the vessels, platelet adherence, myointimal cell proliferation, luminal narrowing and heightened sensitivity of the vessel wall. Clinical evidence of the vessel damage is Raynaud's phenomenon, involving both skin and viscera. The Authors evaluated the effects of iloprost on Raynaud's phenomenon in patients with PSS. This drug provides prolonged vasodilation, reduces platelet aggregation and promotes endothelial lining function repair. This last pattern is of primary importance because it may stop the vicious circle: endothelial injury-platelet hyperaggregation-microangiospasm. Five females were recruited, aged 41-66 years, suffering from well-documented (ARA criteria) PSS, associated with typical Raynaud's phenomenon. The trial provided for intravenous infusion of iloprost at a rate of 1-2 ng/kg/min. First treatment consisted of six-hour infusions on six successive days. After this first treatment, weekly infusions during the winter months were carried on. Drug effectiveness was considered through subjective and objective parameters. All patients showed prominent reduction of number, duration and severity of attacks of Raynaud's phenomenon, improvement of prehensile strength, healing of finger ulcerations and improvement or normalization of digital photoplethysmography. So far, the treatment has been prolonged for years in our patients and still goes on. The side effects of iloprost (headache, flushing, nausea) have been very poor. Therefore, iloprost proved to be a valid drug in the management of Raynaud's phenomenon in patients with PSS, but the inconvenience of intravenous administration may limit its routine use.


Demonstration of transganglionically transported choleragenoid in rat spinal cord by immunofluorescence cytochemistry. Injections of the B-subunit of cholera toxin (CTB) were made into the sciatic nerve of the rat. Following a survival of 2-3 days, the fluorescent antibody technique was used to show that CTB can be utilized as a highly sensitive immunocytochemically detectable transganglionic tracer for primary afferent fibers in the spinal cord. CTB-labeled fibers as well as fibers containing calcitonin gene-related peptide- (CGRP-) or substance P-like immunoreactivity were visualized simultaneously by using different fluorochromes. However, no double labeled fibers were found.


Blockade of nitric oxide synthase inhibits nerve-mediated contraction in the rat small intestine. Inhibitors of nitric oxide (NO) synthase inhibit nerve-mediated non-adrenergic, non-cholinergic (NANC) smooth muscle relaxation in the gastrointestinal tract. In this study, the effect of a NO synthase inhibitor, NG-nitro-L-arginine, was examined on the tetrodotoxin-sensitive NANC contractile response of the rat isolated ileal myenteric plexus-longitudinal muscle evoked by electrical field stimulation. This contraction was concentration-dependently inhibited by NG-nitro-L-arginine (1-3 x 10(-5) M). The inhibition was partly or entirely reversed by L-arginine. The NO synthase inhibitor did not exhibit any non-specific smooth-muscle depressant action or local anaesthetic effect. Sodium nitroprusside, a putative donor of NO also caused a transient contraction of the rat ileal strip. This response was resistant to tetrodotoxin. It is concluded that an NO synthase product is involved in the mechanism of the nerve-mediated NANC primary contraction due to field stimulation in the rat small intestine.


Corticotropin releasing factor and galanin-containing neurons projecting to the median eminence of the rat. To identify corticotropin releasing factor (CRF)- and galanin (GAL)-containing neurons projecting to the median eminence, a retrograde tracing method with True blue was combined with a double-staining method. It was demonstrated that some True blue-labeled CRF neurons in the paraventricular nucleus were also immunoreactive for GAL. These findings support the hypothesis that GAL and CRF are simultaneously involved in regulation of ACTH secretion from the anterior pituitary.


An evaluation of dietary guidance graphic alternatives: the evolution of the eating right pyramid. The pyramid graphic was found to be most effective in conveying the messages of moderation and proportionality. The pyramid design might be further strengthened in conveying the total moderation message by including both FOS pictures and symbols in the same graphic. The pyramid also has considerable strength in promoting the message of variety. More important, the pyramid graphic did not convey misinformation about variety, proportionality, and moderation or about the importance of consuming foods in adequate amounts from all the major food groups to achieve a healthful diet. The bowl design was found to be far less effective in promoting the moderation and proportionality messages, but did illustrate the variety message somewhat better than the pyramid. Although respondents indicated a preference for the bowel shape, it is important to note that even in the groups where preference for the bowl was the strongest (i.e., younger, less educated, ethnic minority, and low-income respondents), the pyramid was more likely to be identified as the graphic that suggested the moderation-of-fat message most clearly. Further, preference for the bowl was not linked to effectiveness of intended messages in those groups. In this study, the shape a respondent preferred was of less importance to policy development than the amount of intended information conveyed by each graphic.


Embryonic stem (ES) cells lacking functional c-jun: consequences for growth and differentiation, AP-1 activity and tumorigenicity. The proto-oncogene c-jun encodes the major component of the transcription factor AP-1 and is thought to have important functions in cell proliferation and differentiation as well as in the cellular response to a variety of external stimuli. To investigate directly the role of c-jun in growth, differentiation and tumorigenicity we generated mouse embryonic stem (ES) cell lines in which both copies of the c-jun gene have been inactivated by homologous recombination. The disruption of both copies of the c-jun gene had no apparent effect on ES cell viability, growth rate and in vitro differentiation potential. Transcriptional activation of the c-jun, junB and c-fos genes following TPA/serum induction was unaffected and efficient transactivation of AP-1 reporter constructs was demonstrated in these cells. Remarkably, subcutaneous injection of ES cells lacking c-Jun into syngeneic mice led to a drastic reduction in the formation of teratocarcinomas. We propose that whereas most of the functions of c-Jun in ES cells appear to be complemented by other Jun proteins in vitro, functional c-Jun protein is essential for efficient tumor growth in vivo.


Platelet activation leads to increased c-src kinase activity and association of c-src with an 85-kDa tyrosine phosphoprotein. We have used platelets as a model system to study the function of c-src in signal transduction and cell adhesion. Numerous proteins were found to be phosphorylated on tyrosine in response to thrombin-induced platelet activation and aggregation. Two phases of phosphorylation were observed, with the second phase, but not the first, being inhibited by blocking platelet aggregation with an Arg-Gly-Asp-Ser tetrapeptide. As a first step towards identifying those proteins phosphorylated on tyrosine and to determine the specific role of p60src during platelet activation, we looked for changes in p60src kinase activity and for associations of p60src with other tyrosine phosphoproteins. The data presented here demonstrate an increase in p60src kinase activity within 1 min of thrombin-induced activation. Furthermore, p60src transiently associates with a tyrosine phosphoprotein during platelet activation and aggregation. This tyrosine phosphoprotein, p80/85, is a previously characterized cytoskeletal substrate for v-src in transformed cells. The data presented here suggest a model in which p60src functions in platelets to link upstream events, such as cell-surface adhesive interactions, with changes in platelet shape and cytoskeletal organization.


Identification and characterization of a cytoskeleton-associated, epidermal growth factor sensitive pp60c-src substrate. In studies aimed at identifying and characterizing pp60c-src substrates that participate in the enhanced mitogenic response to epidermal growth factor (EGF) observed in murine C3H10T1/2 fibroblasts overexpressing c-src, we have identified a 75-kDa protein (p75) whose properties are consistent with those expected of such a substrate. We present evidence to show that p75 is immunologically related to a recently described, cytoskeleton-associated, pp60v-src substrate [Wu et al. (1991). Mol. Cell. Biol., 11, 5113-5124), and that its phosphotyrosine content is increased cooperatively by c-src overexpression and EGF stimulation. p75 is rapidly (within 2 min) phosphorylated on tyrosine upon EGF treatment and undergoes a second, prolonged phase of tyrosyl phosphorylation from 7 to 21 h after EGF addition, suggesting that tyrosyl phosphorylation of p75 is important for late as well as early events following EGF receptor activation. Enhanced tyrosyl phosphorylation of p75 is also seen when cells overexpressing c-src are treated with platelet-derived growth factor (PDGF), but significantly less phosphorylation is observed with insulin and fibroblast growth factor (FGF). Both basal and EGF-induced tyrosyl phosphorylation of p75 are reduced in cells overexpressing mutated forms of c-src (unmyristylated, or kinase deficient) as compared with wild-type c-src overexpressers, indicating the dependence of the enhanced tyrosyl phosphorylation on membrane-associated, enzymatically active pp60c-src. In cellular fractionation experiments p75 partitions with the cytosol, while immunofluorescence studies reveal a striking colocalization with pp60c-src at the plasma membrane and in the perinuclear region. Partial co-staining of p75 and actin occurs at the cell's periphery. These data provide evidence for p75 being a direct substrate of pp60c-src. The possible role of p75 in the enhanced response to EGF seen in c-src overexpressers is discussed.


Overexpressed Drosophila src 64B is phosphorylated at its carboxy-terminal tyrosine, but is not catalytically repressed, in cultured Drosophila cells. Little is known about the regulation of non-receptor tyrosine kinases in invertebrates. We have studied the relationship between the phosphorylation state of the Drosophila src 64B (Dsrc) gene product, p62D, and its tyrosine kinase activity in Drosophila Schneider 2 cells, using wild-type and mutated Dsrc constructs that were overexpressed by transient transfection. Phosphopeptide mapping showed that the putative regulatory C-terminal tyrosine (Tyr-547) of p62D was phosphorylated in vivo. In contrast to vertebrate src family kinases overexpressed in fibroblasts, wild-type p62D overexpressed in Schneider 2 cells was phosphorylated at additional tyrosines outside of the C-terminus. These tyrosines corresponded to the major in vitro autophosphorylation sites. Overexpression of wild-type p62D or several catalytically active p62D mutants significantly increased the phosphorylation of numerous Schneider cell proteins on tyrosine, while expression of catalytically inactive mutants of p62D had no such effect. Thus, in contrast to the repression of src family kinase activity in fibroblasts, p62D is catalytically active when overexpressed in Drosophila cells, perhaps because of substoichiometric C-terminal tyrosine phosphorylation. These results raise the possibility that fly development will be sensitive to ectopic expression of p62D.


Molecular cloning of a family of protein kinase genes expressed in the avian embryo. We have used the polymerase chain reaction (PCR) technique to clone kinase-related sequences from avian blastula, neural crest and neural tube mRNA. Twenty-three distinct protein kinase (PK) sequences were amplified, of which eight are identical to previously described PK genes. The cloned molecules fall into three classes: growth factor receptor tyrosine kinases (RTKs), cytosolic tyrosine kinases and serine/threonine kinases. Among the cloned RTKs were the insulin-like growth factor type I receptor, platelet-derived growth factor receptor alpha, the CEK1 fibroblast growth factor (FGF) receptor as well as the avian homolog of a recently cloned PCR fragment related to the eph/elk/eck family, tyro-5. Furthermore, we cloned a novel FGF receptor-like molecule as well as two novel putative RTKs related to the vascular endothelial growth factor (VEGF) receptor. The pattern of expression of the PCR clones was examined by Northern blot analysis of adult tissues: each molecule recognized one or more transcripts of various sizes, suggesting that PK genes may play regulatory roles both in early development and in adult regulation of tissue function. Together with recent studies, this survey confirms the hypothesis that PKs may play important roles in early vertebrate development.


An Eph-related receptor protein tyrosine kinase gene segmentally expressed in the developing mouse hindbrain. In search of genes possibly involved in the regulation of hindbrain segmentation, we have isolated mouse cDNA clones corresponding to putative protein kinase genes by polymerase chain reaction amplification of cDNA from 9.5-day-old embryo hindbrains. In situ hybridization analysis revealed that one of these genes, Sek, was expressed in an alternating segment-restricted pattern in the developing hindbrain. Isolation and analysis of Sek cDNAs covering the entire coding sequence indicated that Sek encoded a putative receptor protein tyrosine kinase, belonging to the Eph family. These data are consistent with a role of the Sek gene product in a signal transduction process involved in pattern formation in the hindbrain.


Varicella does not appear to be a cofactor for human immunodeficiency virus infection in children. We performed a retrospective analysis of longitudinal clinical and immunologic data obtained from 22 children in the early stages of infection with human immunodeficiency virus (HIV) when they developed varicella. We studied the course of HIV infection to determine whether clinical deterioration occurred after chickenpox. We examined the following indices: growth and development; neurologic status; helper T lymphocyte counts; blood values of core (p24) antigen of HIV; changes in the stage of HIV infection; and need for administration of zidovudine. We studied children for a mean of 2.8 years and for as long as 9.8 years after onset of varicella. There was little evidence that chickenpox affected HIV infection. Three (14%) children developed clinical zoster, 2 of whom (9%) had evidence of chronic infection with varicella-zoster virus. One additional child (5%) had 2 episodes of chickenpox. These observations suggest that children with early HIV infection could be considered for immunization with live attenuated varicella vaccine, which would be predicted to decrease their morbidity from varicella-zoster virus.


Small-conductance chloride channels induced by cAMP, Ca2+, and hypotonicity in HT29 cells: ion selectivity, additivity and stilbene sensitivity. Previous studies in HT29 cells utilizing the cell-attached nystatin (CAN) method [Greger R, Kunzelmann K (1991) Pflügers Arch 419:209-211] have revealed that the Cl- channels induced by cAMP or by increasing cytosolic Ca2+, e.g. by addition of ATP, and by hypotonic cell swelling share in common their conductance, which was so small in our studies [Kunzelmann et al. (1992) Pflügers Arch (in press)] that we could not resolve it at the single-channel level. This prompted the question whether these Cl- conductances can be distinguished in terms of their ion selectivity and sensitivity towards inhibitors. Whether these pathways are additive or not was also examined. The present study utilized the whole-cell patch-clamp and the CAN methods. A total of 160 patches were studied. In whole-cell patches 8-(4-chlorophenylthio)-cAMP (cAMP, 0.1 +/- 1 mmol/l) induced a significant depolarization by 5 mV and a twofold increase in conductance (G) from 6.2 +/- 1.5 nS to 11.7 +/- 3.2 nS (n = 15). Total replacement of Cl- by Br- and I- in cAMP-treated cells hyperpolarized the membrane voltage (V) significantly from -35 +/- 2.8 to -39 +/- 3.4 and -45 +/- 3.3 mV respectively, but had no detectable effect on G, which was 11.9 +/- 3.3 nS in the case of Br- and 11.8 +/- 3.3 nS in the case of I-. Hence, the permselectivity of the cAMP pathway was I- > Br- > Cl-, but the conductances for these anions were all indistinguishable.(ABSTRACT TRUNCATED AT 250 WORDS)


Measurement of cell impedance in frequency domain using discontinuous current clamp and white-noise-modulated current injection. A method is described for the determination of cellular input impedance of non-spiking neurones. The input impedance is important when cellular geometry and the effects of voltage-dependent channels are considered. Cells are impaled with a single glass microelectrod and current is injected using a time-sharing technique. The cell's impedance is measured by randomly modulating the injected current and calculating the impedance as a transfer function between current and recorded membrane voltage. Corresponding coherence functions can also be calculated for estimating the signal-to-noise ratio, and also linearity (i.e. possible activation of voltage-dependent conductances) of the membrane.


Nuclear ion channels in cardiac myocytes. The paradigm that nucleocytoplasmic transport of ions occurs without a diffusional barrier has been challenged by the recent demonstration with patch-clamp techniques of the existence of ion channels in the nuclear envelope of murine zygotes and hepatocytes. This report demonstrates the existence of nuclear ion channels (NIC) in murine ventricular cardiac myocytes. NIC conductance (gamma), calculated from current histogram peaks, was 106-532 pS at 22-36 degrees C. In nucleus-attached patches, replacement of cytoplasmic K+ with Na+ reduced NIC activity within 30 s, suggesting that intranuclear-delimited mechanisms mediate this phenomenon. In excised, inside-out patches K+ was as permeable as Na+ through NIC. NIC activity was observed in 0-4 mM Mg2+ and/or ATP2-, with or without 0-1 mM Ca2+, indicating a minor direct role of these ions. However, in non-responsive excised inside-out patches, NIC activity appeared when the catalytic subunit of the cAMP-dependent protein kinase was applied to the nucleoplasmic side of the patch, in the presence of Mg2+ and ATP2-, indicating an important role for phosphorylation-dependent process(es) in NIC function--an observation supported by the depressing effects of protein kinase inhibitor on responsive NIC. The concept that nucleopore complexes are solely responsible for nucleocytoplamic transport leads to the speculation that these structures are the physical substrate for NIC.


Effects of atrionatriuretic factor on Ca2+ current and Cai-independent transient outward K+ current in human atrial cells. The effect of 10 nM atrial natriuretic peptide (ANF) on macroscopic L-type calcium current, ICa, and calcium-independent outward potassium current, Ilo, were studied in myocytes isolated from human atrial trabeculae using the whole-cell-recording patch-clamp technique. When cells were dialysed with pipette media containing 0.2 mM GTP, ANF reduced ICa by 37.81% +/- 5.4% at +20 mV and Ilo by 21.72% +/- 3.68% at +60 mV in a reversible manner. When ICa was increased by beta-adrenoreceptor stimulation (0.1 microM isoproterenol) or by the phosphodiesterase inhibitor isobutylmethylxanthine (10 microM) ANF reduced ICa by 24.99 +/- 3.4% and by 39.9 +/- 6.3% respectively. In cells dialysed with GTP-free pipette media, ANF increased ICa markedly (39.8% +/- 7%) and reversibly, whereas it still depressed Ilo (18.92% +/- 2%). Addition of 0.2 mM GTP[gamma S] to the pipette solution in the absence of GTP increased ICa, decreased Ilo and suppressed the effect of ANF on both ICa and Ilo. It is suggested that activation of the ANF receptor in human atrial cells reduces ICa via guanylate-cyclase-dependent cGMP production, increases ICa via Gs protein activation and decreases Ilo via Gi protein activation.


The effect of hyperosmotic challenge upon ion transport in cultured renal epithelial layers (MDCK). Exposure of the basal-lateral surfaces of MDCK epithelia, mounted in Ussing chambers, to medium made hyperosmotic by the non-electrolyte mannitol, resulted in a marked inhibition of the adrenaline-stimulated inward short-circuit current (Cl- secretion). This inhibition was unaccompanied by a reversal of the adrenaline-stimulated increment in tissue conductance, indicating that the inhibition was due to modulation of ion transport at the basal-lateral membranes. Loop-diuretic-sensitive 86Rb(K+) efflux mediated by the Na+ - K+ -2 Cl- cotransporter at the basal-lateral membranes was markedly stimulated by hypertonic exposure. A diuretic-sensitive K+ (Cl-) loss was observed in shrunken cells upon prolonged exposure (20 min), showing that the net direction of "cotransport" flux was outward. 86Rb(K+) efflux stimulated by adrenaline (100 microM), exogenous ATP (100 microM) and A23187 (10 microM) was attenuated in shrunken cells, suggesting that basal-lateral K+ conductance is reduced in hyperosmotic media. "Cotransport" stimulation by hyperosmotic medium was asymmetric, apical bathing hypertonicity being ineffective. These data are consistent with a low hydraulic permeability of the apical membranes.


Evolutionary conserved nucleotides within the E.coli 4.5S RNA are required for association with P48 in vitro and for optimal function in vivo. E.coli 4.5S RNA is homologous to domain IV of eukaryotic SPR7S RNA, the RNA component of the signal recognition particle. The 4.5S RNA is associated in vivo with a 48kD protein (P48), which is homologous to a protein component of the signal recognition particle, SRP54. In addition to secondary structural features, a number of nucleotides are conserved between the 4.5S RNA and domain IV of all other characterised SRP-like RNAs from eubacteria, arachaebacteria and eukaryotes. This domain consists of an extended stem-loop structure; conserved nucleotides lie within the terminal loop and within single-stranded regions bulged from the stem immediately preceding the loop. This conserved region is a candidate for the SRP54/P48 binding site. To determine the functional importance of this region within the 4.5S RNA, mutations were introduced into the 4.5S RNA coding sequence. Mutated alleles were tested for their function in vivo and for the ability of the corresponding RNAs to bind P48 in vitro. Single point mutations in conserved nucleotides within the terminal tetranucleotide loop do not affect P48 binding in vitro and produce only slight growth defects. This suggests that the sequence of the loop may be important for the structure of the molecule rather than for specific interactions with P48. On the other hand, nucleotides within the single-stranded regions bulged from the stem were found to be important both for the binding of P48 to the RNA and for optimal function of the RNA in vivo.


Cleavage efficiencies of model substrates for ribonuclease P from Escherichia coli and Thermus thermophilus. We compared cleavage efficiencies of mono-molecular and bipartite model RNAs as substrates for RNase P RNAs (M1 RNAs) and holoenzymes from E. coli and Thermus thermophilus, an extreme thermophilic eubacterium. Acceptor stem and T arm of pre-tRNA substrates are essential recognition elements for both enzymes. Impairing coaxial stacking of acceptor and T stems and omitting the T loop led to reduced cleavage efficiencies. Small model substrates were less efficiently cleaved by M1 RNA and RNase P from T. thermophilus than by the corresponding E. coli activities. Competition kinetics and gel retardation studies showed that truncated tRNA substrates are less tightly bound by RNase P and M1 RNA from both bacteria. Our data further indicate that (pre-)tRNA interacts stronger with E. coli than T. thermophilus M1 RNA. Thus, low cleavage efficiencies of truncated model substrates by T. thermophilus RNase P or M1 RNA could be explained by a critical loss of important contact points between enzyme and substrate. In addition, acceptor stem--T arm substrates, composed of two synthetic RNA fragments, have been designed to mimic internal cleavage of any target RNA molecule available for base pairing.


Amplification of protein expression in a cell free system. Large quantities of a catalytically active protein have been produced in a cell free system. More than 10(9) copies of protein were produced from each DNA plasmid containing DNAfol, the bacterial gene encoding dihydrofolate reductase (DHFR). The strategy employed, denoted gene amplification with transcription/translation (GATT), involves sequential coupling of (i) DNA amplification by the polymerase chain reaction (PCR) and (ii) in vitro RNA transcription by T7 RNA polymerase, followed by (iii) translation of the run-off transcripts in a rabbit reticulocyte system. The protein product had the expected size (18 kDa) and catalyzed the NADPH-dependent reduction of 7,8-dihydrofolic acid to 5,6,7,8-tetrahydrofolic acid as efficiently as authentic DHFR. Potential applications of the strategy include large scale production of enzymes containing synthetic amino acids and facilitation of the characterization of the function of genes encountered in genomic mapping studies.


In vitro effect of antisense oligonucleotides on human immunodeficiency virus type 1 reverse transcription. The molecular events involved in antisense-mediated inhibition of retroviral transcription were studied by analyzing the in vitro effect of antisense oligodeoxynucleotides on reverse transcription by Human Immunodeficiency Virus type 1 (HIV-1) reverse transcriptase (RT). Oligonucleotides have been designed to be complementary to three targets located in the 5' region of the HIV-1 RNA genome: the transactivating response element (TAR), the U5 region and a sequence contiguous to the primer binding site (PrePBS). Antisense oligodeoxynucleotides were used with their 3'-OH end either free or blocked by a dideoxynucleotide in order to avoid cDNA synthesis. Experiments with two recombinant forms of HIV RT, carrying or not RNase H activity, showed that antisense oligonucleotides can arrest reverse transcription by an RNase H-independent mechanism. The AntiTAR oligonucleotide did not affect reverse transcription. In contrast, the AntiU5 and AntiPrePBS oligonucleotides led to an efficient inhibition of both forms of HIV RT. In the case of the AntiU5, the inhibition obtained in the absence of the RNase H activity indicates that this effect can be related to features of the RNA secondary structure. The AntiPrePBS oligonucleotide did bind to its target only in the presence of PBS primer. Use of shifted oligonucleotides showed that the AntiPrePBS inhibitory effect depends on a cooperative annealing with the AntiPBS primer on the template.


Inhibition of 3 alpha-hydroxysteroid oxidoreductase and 5 alpha-reductase activity by antiandrogens and indomethacin in the rat prostate. We studied the effects of antiandrogens in vitro on inhibition of 3 alpha-hydroxysteroid oxidoreductase (3 alpha-HSOR) and 5 alpha-reductase activities in the rat prostate. Kinetic and inhibition experiments were analyzed by thin layer chromatography. Cyproterone acetate (CA), chlormadinone acetate (CMA), and TZP-4238 (TZP), which is more potent than other antiandrogens, were used as inhibitors and were compared with indomethacin IND, which is a recognized 3 alpha-HSOR inhibitor. The IC50S of CA, CMA, IND, and TZP for 3 alpha-HSOR reductase in cytosol were about 5, 10, 10, and 100 microM, respectively, and inhibition was competitive. The IC50 of IND for 3 alpha-HSOR reductase in microsomes was 20 microM. The IC50S of other inhibitors were > 100 microM, and inhibition was noncompetitive. The IC50S of CA, CMA, IND, and TZP for 3 alpha-HSOR oxidase in cytosol were > 100 microM, and inhibition was competitive or noncompetitive. Inhibition of 3 alpha-HSOR oxidation was not observed in microsomes. The difference between these inhibition patterns suggests that there may be 4 isoenzymes in rat prostatic tissue. The IC50S of MK-906, CMA, and TZP for 5 alpha-reductase in prostate homogenate were about 0.01, 200, and 200 microM, respectively.


Measurement of androgen sensitivity in the human prostate in in vitro three-dimensional histoculture. We have adopted an in vitro three-dimensional histoculture technique for assay of androgen sensitivity in explants of human benign prostatic tissue. The assay is based on the uptake of 3H-thymidine/micrograms protein in explants of prostate incubated in parallel with dihydrotestosterone (DHT) and hydroxyflutamide (HF) controls. The ratio of 3H-thymidine/micrograms protein in DHT treated samples per 3H-thymidine/micrograms protein in HF treated samples provides an index of androgen sensitivity. The DHT/HF index measured in 24 BPH specimens averaged 3.6. To determine the specificity of the HF effect, we measured the DHT/HF index in a single prostate at different concentrations of HF in the presence of fixed concentrations of DHT (2 x 10(-8) M) and noted a dose-response relationship. In addition we noted no effects of HF on 3H-thymidine incorporation over a range of 2 x 10(-4)M compared to 2 x 10(-7)M, except at the highest concentration. Of surprise was the finding of an average DHT/HF index in 5 different nonprostate tissues, including breast, uterus, colon, kidney, and thyroid, that was similar to the index found in prostates. We plan to adapt this androgen sensitivity assay to measure the DHT/HF index in biopsy-size samples of prostate, since such an assay could then be utilized to determine androgen sensitivity in individual patients with prostate cancer.


Lymphatic drainage of the prostate gland in canines. Lymphography of the penis was performed in 3 canines, and direct intraprostatic injection of India ink was carried out in 8 canines to visualize lymphatic drainage of the prostate gland. In penile lymphography, the contrast medium drained in 2 directions: (1) lymphatic drainage into the external and common iliac lymph nodes through the superficial inguinal lymph nodes; (2) lymphatic drainage into the presacral lymph nodes along the urethra and posterior wall of the pelvis. Lymphatics of the prostate gland drained in 3 directions: (1) lymphatics from the prostate gland mainly drained along the prostatic and internal iliac vessels into the internal and common iliac lymph nodes, (2) lymphatics from the dorsal region near the urinary bladder drained along the ureter into the common iliac lymph nodes, and (3) lymphatics from the apex of the prostate gland drained along the posterior wall of the pelvis into the presacral lymph nodes. Prostate cancer developed at the apex is estimated to preferentially metastasize to the presacral lymph nodes, and drug injection into the penile lymphatics is considered to be a good route to treat metastases of lymph nodes in the pelvis.


The response to alpha blockade in benign prostatic hyperplasia is related to the percent area density of prostate smooth muscle. The objective of the present study was to determine whether the smooth muscle content of the prostate adenoma is related to the clinical response to terazosin, a long-acting selective alpha 1 blocker. Multiple random biopsies of the prostate were obtained from 26 male subjects with symptomatic benign prostatic hyperplasia (BPH) prior to initiating therapy with terazosin. Double immunoenzymatic staining and computer-assisted quantitative color image analysis were utilized to quantify the area density of smooth muscle, connective tissue, glandular epithelium, and glandular lumen. The clinical response to alpha blockade was based upon changes in peak urinary flow rate and the Boyarsky symptom score. A significant direct relationship was observed between the percent area density of smooth muscle and the percent change in peak urinary flow rate. A statistically significant correlation between the percent area density of smooth muscle and the percent change in Boyarsky symptom score was not observed. The percent area density of prostate smooth muscle in the subjects exhibiting a favorable clinical response was 38% greater than the nonresponders (P = 0.068). The clinical response to alpha blockade in BPH is related to the area density of prostate smooth muscle.


Immunohistochemical evidence of the existence and localization of aromatase in human prostatic tissues. Estrogens may be involved in normal growth of the prostate and the development of benign prostatic hyperplasia (BPH). The location of estrogen production is still unclear, and there has never been a direct evidence for the existence of the aromatase system, which converts androgens to estrogens, in the prostate. Using an avidin-biotin technique with a polyclonal anti-human placental aromatase, we demonstrated the existence of aromatase in normal prostates of young men and BPH tissue from elderly men. The staining is more pronounced in the stroma. However, positive stains were also seen in the glandular epithelium. While evidence of the existence of an enzyme system does not equal demonstration of its activity in a specific tissue site, our findings suggest that local estrogen production in the stroma and/or epithelium of the prostate may play a role in the maintenance of normal growth and development of BPH.


Effects of the new steroidal antiandrogen TZP-4238 on hormone-induced canine prostatic hyperplasia. The effects of the new steroidal antiandrogen TZP-4238 on hormone-induced canine prostatic hyperplasia (BPH) were studied in comparison with those of chlormadinone acetate (CMA), a steroidal antiandrogen used in Japan. One- to 2-year-old beagle dogs were castrated and administered 75 mg/week of androstanediol (A-diol) plus 0.75 mg/week of estradiol (E2) for 25 weeks. These dogs were treated orally with placebo, 0.5 mg/kg/day of TZP-4238, 0.1 mg/kg/day of TZP-4238, and 2.5 mg/kg/day of CMA, respectively, for 21 weeks after 4 weeks treatment with A-diol plus E2. Treatment with 0.5 mg/kg/day of TZP-4238 or 2.5 mg/kg/day of CMA suppressed prostatic growth, and treatment with 0.1 mg/kg/day of TZP-4238 suppressed prostatic growth slightly. Treatment with 0.5 mg/kg/day of TZP-4238 decreased 5 alpha-reductase activity, DHT content, and nuclear androgen receptor (AR) content in the prostate, and treatment with 0.1 mg/kg/day of TZP-4238 or 2.5 mg/kg/day of CMA also decreased or tended to decrease these parameters. In conclusion, TZP-4238 and CMA were effective in inhibiting the growth of hormone-induced canine BPH, and TZP-4238 was at least 5 times more potent than CMA. TZP-4238 inhibited prostatic growth by decreasing prostatic androgen content and the androgen-AR complex. TZP-4238 decreased 5 alpha-reductase activity by prevention of the androgen action described above.


Pore size and negative charge as structural determinants of permeability in the Torpedo nicotinic acetylcholine receptor channel. To gain an insight into the molecular basis of ion permeation mechanism through the nicotinic acetylcholine receptor (AChR) channel, we have determined permeability ratios of organic cations relative to Na+ of specifically mutated Torpedo californica AChR channels expressed in Xenopus oocytes. The mutations involved mainly the side chains of the amino acid residues in the intermediate ring, where mutations have been found to exert strong effects on single-channel conductance and ion selectivity among alkali metal cations. The results obtained reveal that both the size and the net charge of the side chains of the intermediate ring are involved in determining the permeability, and provide experimental evidence that the pore size at the intermediate ring is a critical determinant of permeability. Our findings further suggest that changes in net charge exert effects on permeability by affecting the pore size of the channel.


Involvement of prostanoids in the regulation of angiogenesis by polypeptide growth factors. Polypeptide growth factors (PGFs), mainly those of the fibroblast growth factor (FGF) family, have been shown to be capable of regulating angiogenesis. Although many data have been accumulated during this last year on the mechanism of action of PGF, little is known about a possible identification of second messengers signalling to the cell the occupancy of the receptor by its ligand. We have previously proposed that arachidonic acid or its derivatives may play a role as PGF second messengers. In the present paper we described a modification of the chorioallanthoic membrane (CAM) technique, involving the use of labelled sulphate to follow the angiogenic process. Thus we have been able to correlate morphological observation of CAMs development with incorporation of labelled sulphate in a stable form. Here we show that, as expected, PGF as endothelial cell growth factor (ECGS) or basic fibroblast growth factor (bFGF) potentiate the incorporation of radioactivity into CAMs at concentrations which for bFGF are of the order of 1.5 micrograms/egg. This effect can be correlated to the generation of prostanoids by two kinds of approach: A) PGE1 injected into eggs was capable of strongly increasing labelling of CAMs; B) Indomethacin had a dramatic effect on embryo survival as well as on CAM development, decreasing both at very low concentration (50 survival rate observable at 2 micrograms/egg). Finally vanadate, which is known to inhibit tyrosine phosphatase, was capable of potentiating the effect of PGF on angiogenesis. Thus it appears that products of the prostaglandin H synthase pathway behave as mediators of PGF control of angiogenesis.


Convergent and overlapping transcripts of the Chlamydia trachomatis 7.5-kb plasmid. Transcription of the 7.5-kb cryptic plasmid of Chlamydia trachomatis serovar L2 was investigated. Faint, diffuse transcripts of about 1.6 and 2.2 kb and intense short transcripts of about 250 and 430 bases were identified by Northern blot analysis. The short transcripts were found to have a common 5' end corresponding to bp 501 relative to the unique BamHI site of the plasmid and to terminate at different downstream sites. Putative promoter sequences of TTGCCA and TATATT, which closely resemble the consensus recognition site of Escherichia coli sigma 70, were identified at the -35 and -10 positions upstream from the 5' end of the short transcripts made in chlamydia. Transcripts of similar sizes were also expressed from this promoter in E. coli harboring a recombinant plasmid encoding the short transcripts. The short transcripts encode a common open reading frame (ORF) of 34 codons; however, a strong ribosome binding site was not found in the vicinity of the initiator codon, and it is not known whether the transcripts are translated in vivo. A large ORF of 330 codons, which has been shown to encode a hypothetical protein containing conserved domains of recombinase-like proteins, is antisense to the short transcripts. Transcripts encoding the large ORF could not be detected directly by Northern blot or primer extension analysis. However, transcripts were detected by polymerase chain reaction amplification of the large ORF cDNA and when Southern blots of single-stranded antisense DNA for the large ORF were probed with radiolabeled RNA synthesized by host-free chlamydial reticulate bodies. Thus, both strands of the chlamydial plasmid are transcribed in the region encoding the short transcripts. We propose that the short transcripts play a regulatory role as antisense RNAs.


[The epidemiology of adverse reactions to nonsteroidal anti-inflammatory drugs. Nimesulide is tolerated by patients with adverse reactions to NSAIDs and modulates in man the skin response to histamine and codeine]. We evaluated the tolerance to NIM in patients with adverse reactions to NSAIDs, expressed by urticaria-angioedema, bronchial asthma or erythema polymorphous on 92 patients among which 32 atopics affected by asthma, rhinitis or contact dermatitis. Challenge test has been performed with increasing amounts of NIM per os every 2 hrs to a maximum of 100 mg/dose. The last dosage was repeated every 12 hrs for 4 times more. No reactions were observed in 86 patients (93.4%). Moreover we evaluated the efficacy of NIM in modulating pomphoid cutaneous reaction to histamine (HIS) and codeine (COD). Three subjects were prick tested with HIS and COD solutions at different concentrations (0.1 to 10 mg/ml), before and after a 5 days therapy with NIM at different dosages/kg/die. At NIM dosages of 3.7 and 2.7 mg/kg/die we observed a strong reduction of pomphoid activity of both HIS and COD. No clear effects were detected at 2.2 mg/kg/die dosage. We assume a dose-related in vivo inhibitory effect of NIM on cutaneous mast cells releasability or a direct anti-histaminic effect. We point out the possible therapeutic role of NIM in the treatment of allergic flogosis at steroid and cromon's side.


Metabolism of FK 506 in differentially induced rat liver microsomes. The in vitro hepatic metabolism of FK 506 was studied in microsomes prepared from control rats as well as in microsomes prepared from rats treated with the selective cytochrome P-450 isozyme inducers 3-methylcholanthrene (IA), phenobarbital (IIB), and dexamethasone (IIIA). The metabolism of FK 506 was similar for control microsomes and for microsomes prepared from phenobarbital and 3-methylcholanthrene induced animals. The percentage of FK 506 metabolized by these tissue preparations ranged from 21.7 to 32.7%. In contrast, the percentage of FK 506 metabolized by dexamethasone induced microsomes was 86.4%. The metabolism of FK 506 was not effected when the selective IA and IIB isozyme inhibitors alpha-naphthoflavone and orphenadrine were added to the incubations. However, the metabolism of FK 506 decreased by approximately 44% when the IIIA specific isozyme inhibitor troleandomycin was added to the dexamethasone induced microsomes. Therefore, the metabolism of FK 506 is apparently mediated primarily by the steroid inducible cytochrome P-450 IIIA isozyme.


The effect of ruthenium red during Ca2+ depletion and repletion in the isolated perfused rat liver. Perfusion of rat liver with Ca(2+)-depleted buffer induces oxidative stress and liver damage, which can be prevented by Ca2+ repletion (Okuda et al. J Lab Clin Med). In the present study, we investigated the action of ruthenium red on acute Ca2+ loading after Ca2+ depletion in the isolated perfused rat liver. The major findings of this study are that 1) Ca2+ depletion-induced liver damage was related to mitochondrial disfunction; 2) ruthenium red inhibited the oxidative stress and liver damage normally seen during Ca2+ depletion; 3) ruthenium red inhibited the Ca2+ depletion-induced mitochondrial disfunction. These observations suggest that mitochondrial Ca2+ cycling is responsible for Ca2+ depletion-induced oxidative stress and liver damage.


Restricted expression of immunoglobulin light chain mRNA and of the adhesion molecule CD11b on circulating monoclonal B lineage cells in peripheral blood of myeloma patients. Circulating monoclonal B cells in peripheral blood from patients with multiple myeloma or with monoclonal gammopathy of undetermined significance (MGUS) have previously been shown to express CD19, CD20, and PCA-1 and are predominantly CD45R0+, characterizing them as very late stage B cells. This work shows that the abnormal B cells are monoclonal as defined by their exclusive expression of either kappa or lambda light chain mRNA, and that the same type of light chain mRNA is expressed in both bone marrow plasma cells and blood B cells. These abnormal tumour-related circulating B cells express high densities of CD11b, a beta 2-integrin, which is expressed in a conformationally active state as defined by reactivity with monoclonal antibody 7E3. Normal peripheral blood B cells which do not bear CD11b acquire a high density after stimulation with pokeweed mitogen (PWM). At day 4 of culture, the expression of CD11b on normal CD19+ B cells was nearly comparable to that of the circulating myeloma late stage B cells. After PWM stimulation of circulating myeloma B cells the expression of CD11b was gradually lost during 4 days of culture, suggesting that its expression is dynamically regulated. Two patients with no phenotypically abnormal B cells in their blood at diagnosis acquired a large subset of CD11b+ B cells 4 weeks after initiation of chemotherapy. In most patients, a subset of the circulating myeloma B cells express a low density of CD5. The proportion of CD19+ B cells in the bone marrow expressing CD11b was much reduced compared with peripheral blood B cells, and CD11b was not detectable on plasma cells in the bone marrow, suggesting a sequential relationship of the B-cell subsets detected in our population of patients, involving gradual loss of CD11b concurrent with the loss of CD19 during B lineage differentiation. These cells appear to represent a continuously differentiating monoclonal B lineage culminating in the CD11b- plasma cell entrenched in the bone marrow. We speculate that the expression of conformationally active CD11b on the abnormal B cells in peripheral blood mononuclear cells of myeloma patients facilitates transendothelial migration of circulating myeloma B cells to the bone marrow.


Complement receptor type 2 mediates infection of the human CD4-negative Raji B-cell line with opsonized HIV. Opsonization of the HTLV-RF and HTLV-IIIB strains of HIV-1 with normal human HIV seronegative serum under conditions that allow complement activation resulted in the productive infection of cells of the Raji B lymphoblastoid cell line. Under the same experimental conditions, no infection of Raji cells was observed with unopsonized virus. Infection of Raji cells with complement-opsonized HIV-1 was totally suppressed by preblocking the function of CR2 (the C3dg receptor, CD21) on the cells with a monoclonal anti-CR2 antibody cross-linked with rabbit F(ab')2 anti-mouse immunoglobulin antibodies. Infection of Raji cells occurred independently of CD4 since the cells lacked the expression of CD4 antigen and of CD4 transcripts. Thus, Raji cells may be infected with complement-opsonized HIV independently of CD4 and in the absence of antibodies. By mediating and/or enhancing HIV infection, complement and complement receptors contribute to extend the range of target cells to the virus and may increase infection in patients with a low viral load.


Visualizing cells in three dimensions using confocal microscopy, image reconstruction and isosurface rendering: application to glial cells in mouse central nervous system. This paper describes a general method for visualizing individual cells in intact tissue in three dimensions. The method involves immunostaining intact tissue to label specific cells, "optical sectioning" the stained tissue by laser scanning confocal microscopy, computationally reconstructing a three dimensional image data set from the digitized confocal optical sections, delineating isosurfaces of specific intensity within the reconstructed image by a "marching cubes" algorithm to generate polygon meshes defining boundaries of cells, and displaying individual cells, identified as three dimensional objects enclosed by contiguous polygon meshes, using computer graphics techniques. Each of the components of this method has been described previously in conjunction with other applications. However the combination of these techniques to visualize a variety of different individual cell types in three dimensions in intact tissue represents a new approach. To illustrate the application of this method, we have visualized three different glial cell types in mouse CNS tissue. Oligodendrocytes, specifically stained with antibody to myelin basic protein, were used as an example of cells labelled with an internal membrane antigen. Astrocytes, specifically stained with antibody to glial fibrillary acidic protein, were used as an example of cells labelled with a cytoplasmic antigen. Microglia, specifically stained with Mac.1 antibody, were used as an example of cells labelled with an external membrane antigen. The images that are generated contain remarkably detailed volumetric and textural information that is not obtainable by conventional imaging techniques.


[Retrospective survey of pharmacologic tolerance in the prevention of neoplastic recurrence of superficial urothelioma]. As from June 1989 all superficial bladder tumours (regardless whether treated by surgery or endoscopy) have been included in 5 different chemo, immuno or chemoimmunoprophylactic protocols of therapy against neoplastic recurrence. The protocols considered: mitomycin alone and in association with alpha 2a IFN; epirubicin alone and in association with alpha 2b IFN; immunomodulating alpha 2a IFN. The administration procedures adhered to those set down in the literature and late treatment modalities were used for the associations. The aim of the study was to perform a retrospective evaluation of the intravesical therapies regarding local and systemic tolerability to various drugs and disregarding any evaluation of drug efficacy, i.e. the percentage of patients free from recurrence. We evaluated 92 patients and a total of 1028 instillations carried out prior to June 1991. The number of patients in each protocol was: 31 on IFN; 13 on mitomycin; 24 on epirubicin; 14 on the immunomodulator plus with mitomycin; 10 on the immunomodulator associated with epirubicin. Treatment was suspended in 11/92 patients (11.9%). The highest percentage of interruptions (21.4%) was observed in the mitomycin plus IFN protocol, while the lowest (6.4%) was seen in the IFN monoprotocol. The percentage of patients who presented no side effects during prophylaxis was similar in all protocols studied (50% of patients on the two associations and 54.8% on IFN alone) with the exception of the epirubicin protocol patients who showed lower tolerance (41.6%). The number of "irritable" instillations in each protocol group was analyzed. The lowest number (9/162 = 5.5%) was conserved in the epirubicin plus IFN protocol, while the highest (19/108 17.5%) was seen in the mitomycin group.(ABSTRACT TRUNCATED AT 250 WORDS)


[New methods for the evaluation of cervico-prostatic obstruction]. To study outflow obstructions Schafer (1989) has recently proposed a simplified method based on a graph derived from over 1000 pre and post-op. P/F studies. 42 patients with BPH were evaluated according to this method. 11 patients had no outlet obstruction (26%). Surprisingly, only 15 had a normal detrusor, while 27 had a weak detrusor. The second part of the study concerns 24 patients with post-prostatectomy frequency-dysuria syndrome. Only 21% of these patients showed an outlet obstruction; while 58% showed a weak detrusor as responsible for their symptoms.


Posterior decompression and stabilization for multiple metastatic tumors of the spine. Although the value of surgical decompression and stabilization for solitary spinal metastasis is well documented, indication for surgery for advanced multiple metastatic tumors of the spine is controversial. In this study, the clinical effect of posterior decompression and stabilization was investigated in 11 patients with advanced multiple spinal metastases with unfavorable conditions. Mean blood loss during surgery was 3000 g. Disseminated intravascular coagulation occurred in three patients. Neurologic improvement was observed in nine patients. There was no neurologic deterioration due to surgery in any patients. A measure of pain relief was obtained in all patients. However, the postoperative longevity was short and the patients died 2.5 months (on average) after operation, except in cases of breast cancer. The effect of the posterior surgery on multiple spinal metastases depended on primary diseases. In cases of short life expectancy, the effect of the surgery was limited only to the short duration of neurologic improvement, pain relief, and ease of nursing care while confronted with grave surgical morbidity. In cases of long life expectancy with tumors like breast cancer, however, posterior decompression and stabilization were expected to exert long-term therapeutic effect. Therefore, the posterior surgery for multiple spinal metastases is cautiously indicated considering the nature of the primary tumor.


[Half-body irradiation. An effective palliative treatment of widespread skeletal metastases]. In a large number of cancer patients, extensive skeletal metastases or myelomatosis induce vast suffering, such as intolerable pain and local complications of neoplastic bone destruction. Analgetic drugs frequently do not yield sufficient palliation. Irradiation of local fields often has to be repeated, because of tumour growth outside previously irradiated volumes. Wide field irradiation of the lower or upper half of the body causes significant relief of pain in most patients. Adequate pretreatment handling of patients, method of irradiation, and follow-up are of marked importance to reduce side effects, and are described as they are carried out at the Department of Oncology, Haukeland Hospital.


Subacute ethanol consumption reverses p-xylene-induced decreases in axonal transport. Human exposure to organic solvents is often complicated by ethanol ingestion and the literature is replete with demonstrations of metabolic interactions between ethanol and organic solvents at a pharmacokinetic level. Because of the possible modulation of xylene toxicity by ethanol consumption, the present group of studies characterizes the effect of ethanol on the p-xylene-induced decrease in axonal transport in the rat optic system previously reported by our laboratory. Long-Evans, hooded, male rats were divided randomly into two groups: those receiving 10% ethanol in their drinking water and those receiving water only. These two groups were further subdivided into two groups which were either exposed by inhalation to 1600 ppm p-xylene for 6 h/day, 5 days/week for 8 exposure-days or were treated identically except that they were exposed to air while in the inhalation chambers. The ethanol-drinking rats were given ethanol 6 days prior to and on the days of the inhalation exposure. Immediately after removal from the inhalation chambers on the last exposure day, the animals were injected intraocularly with [35S]methionine and [3H]fucose to measure the synthesis and rapid axonal transport of proteins and glycoproteins, respectively, in the retinal ganglion cells. The animals were sacrificed 20 h later, and the amount of radioactivity in different areas of the retinal ganglion cells was determined by liquid scintillation counting. As in previous experiments, the xylene exposure group showed a significant reduction in axonal transport of proteins and glycoproteins, whereas the ethanol exposure alone produced no significant reductions in the transport of either proteins or glycoproteins. In the animals receiving both ethanol and xylene, however, the ethanol treatment prevented the decreased transport characteristic of the xylene only animals, i.e. in all areas of the optic projections the level of transport were similar to the level present in the control groups. These data suggest that the xylene-induced reduction in rapid axonal transport was reversed (or prevented) by subacute ethanol consumption.


Effects of feeding and fasting on hepatolobular distribution of glutathione and cadmium-induced hepatotoxicity. Relationship between hepatolobular distribution profile of glutathione (GSH) and cadmium (Cd)-induced hepatotoxicity was examined in both fed and fasted rats by computerized densitometry of histochemically stained GSH in the liver sections using an image analyzer system. In fed rats, density gradient distribution of hepatolobular GSH, which was higher in the periportal region than in the perivenous one, was always observed even at a diurnally minimal concentration of GSH. This heterogeneous distribution of GSH, however, disappeared in fasted rats, even though the hepatic GSH concentration recovered to 81% of the control level in rats fasted for 48 h. In histopathological examination on livers 24 h after oral treatment of fed and fasted rats with 60 mg Cd/kg, zonal necrotic changes were observed from the perivenous to midlobular region but not in the periportal one in fed rats even at a diurnally minimal concentration of hepatic GSH. On the other hand, necrotic changes in the liver extended to the panlobular region including the periportal one in fasted rats. These necrotic changes were greater with a longer duration of fasting. These results suggest that the density gradient distribution of hepatic GSH but not the actual concentration of the compound plays an important role in protecting rats against acute hepatotoxicity of Cd.


Evaluation of surfactant cytotoxicity potential by primary cultures of ocular tissues: I. Characterization of rabbit corneal epithelial cells and initial injury and delayed toxicity studies. This investigation was undertaken to develop cytotoxicity assay systems using primary cultures of rabbit corneal epithelial cells as an experimental model to evaluate oculotoxic agents and the ability of these in vitro assay systems to predict irritancy potential and delayed toxicity. We have characterized the epithelial nature of the cultures by identifying keratins with antikeratin antibodies (AE1/AE3) and by demonstrating metabolic enzymes important to the integrity of the cells: lactate dehydrogenase, glucose 6-phosphate dehydrogenase and aldolase. Eight surfactants were compared and ranked according to their cytotoxic potential. We evaluated cytotoxicity by measuring leakage of the cytosolic enzyme, lactate dehydrogenase, into the medium, by making morphological observations and by assessing lysosomal neutral red uptake and mitochondrial 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction. The cells were treated for 1 h with the surfactants and the possibility of delayed toxicity was evaluated 24 h after removal of the surfactant. The cytotoxicity of the different types of surfactants as shown by all the tests was cationic > anionic = amphoteric > non-ionic. Triton X-100, a non-ionic surfactant but a severe irritant, had a ranking similar to anionic surfactants. The in vitro rankings corresponded well to reported in vivo Draize rabbit eye test data. The 24-h test for lactate dehydrogenase leakage showed that mild and non-irritating surfactants did not demonstrate any subsequent damage after a 1-h exposure, but the extreme and severe surfactants continued to show further damage after the 1-h exposure. These in vitro findings were similar to reported in vivo results. The neutral red and MTT tests did not adequately predict the prolonged toxicity of the more irritating surfactants, as was demonstrated by the lactate dehydrogenase leakage test. We conclude that in vitro cytotoxicity assays using primary cultures of rabbit corneal epithelial cells may be used to rank the cytotoxic potential of surfactants, but only the lactate dehydrogenase leakage test was able to assess prolonged cell injury.


Differences in alpha 2u-globulins increased in male rat kidneys following treatment with several alpha 2u-globulin accumulating agents: cystein protease(s) play(s) an important role in production of kidney-type-alpha 2u-globulin. Effects of alpha 2u-globulin accumulating agents on alpha 2u-globulins in rat kidneys were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting analysis. Treatment of male animals with decalin (150 mg/kg), 2,2,4-trimethylpentane (50 mg/kg), isophorone (150 mg/kg), d-limonene (150 mg/kg) or 1,4-dichlorobenzene (150 mg/kg) by gavage for 14 consecutive days in each case resulted in a marked intensification of a protein band corresponding to kidney-type-alpha 2u-globulin, with a molecular mass calculated to be approximately 16 kDa. However, intraperitoneal treatment with leupeptin and E-64 (two times 0.07 mmol/kg, for each), well known cystein protease inhibitors, while only slightly increasing this kidney-type-alpha 2u-globulin band, caused the intensification of a approximately 19-kDa molecular mass protein band which was revealed to be a native-type-alpha 2u-globulin by SDS-PAGE and immunoblotting. These results indicated that at least two types of alpha 2u-globulin can be increased in male rat kidney by chemical treatment. Moreover, cystein protease(s) appear(s) to play an important role in the degradation of alpha 2u-globulin and particularly in the conversion of native-type-alpha 2u-globulin to kidney-type-alpha 2u-globulin in rat kidneys.


Single neurone models: oversimple, complex and reduced. The single neurone stands in the midst of a controversy among modelers. Some believe that its details are functionally superfluous when the neurone operates in a large network, and very simple models can be used to represent the input/output characteristics of neurones. Others claim that the unique morphology and electrical properties of neurones do play an important role. Complicated models of neurones are developed to reveal how the various kinds of 'neurone-ware' (dendrites, spines, axons, membrane channels and synapses) create a computationally powerful unit. Various models are discussed, including new carefully reduced models that retain essential features of more complex models. Such intermediate models will play a central role in our efforts to understand information processing in large neuronal networks.


Reconstruction of neuronal networks in culture. Since the 1960s, the large neurones of some invertebrates have been exploited in attempts to define the neural circuits that underlie simple behaviours. Even in the relatively 'simple' nervous systems of these animals, it is often difficult to study individual synaptic connections in detail and to rule out involvement of unidentified neurones. These limitations have been overcome by reconstruction of partial circuits of identified neurones in cell culture. This approach has provided opportunities to examine the function of small neuronal circuits in a manner that is unapproachable in the intact nervous system.


Reliability and information transmission in spiking neurons. Spiking neurons encode continuous, time-varying signals in sequences of identical action potentials. Relatively simple algorithms allow one to 'decode' this neural representation of sensory data to estimate the input signals. Decoding experiments provide a quantitative characterization of information transmission and computational reliability under real-time conditions. The results of these studies show that neural coding and computation in several systems approach fundamental physical and informational theoretic limits to performance.


Modelling of intersegmental coordination in the lamprey central pattern generator for locomotion. Rhythmic motor activity requires coordination of different muscles or muscle groups so that they are all active with the same cycle duration and appropriate phase relationships. The neural mechanisms for such phase coupling in vertebrate locomotion are not known. Swimming in the lamprey is accomplished by the generation of a travelling wave of body curvature in which the phase coupling between segments is so controlled as to give approximately one full wavelength on the body at any swimming speed. This article reviews work that has combined mathematical analysis, biological experimentation and computer simulation to provide a conceptual framework within which intersegmental coordination can be investigated. Evidence is provided to suggest that in the lamprey, ascending coupling is dominant over descending coupling and controls the intersegmental phase lag during locomotion. The significance of long-range intersegmental coupling is also discussed.


Motor-pattern-generating networks in invertebrates: modeling our way toward understanding. Motor-pattern-generating networks in invertebrates have been the objects of intensive study to determine the origin and modulation of rhythmic neural activity. In some pattern generators, intrinsically bursting neurons drive activity throughout the network. In most pattern generators, however, rhythmicity arises from the interplay between intrinsic membrane properties and synaptic interaction. Reciprocal inhibitory synapses between neurons are thought to be crucial for generating oscillation in these networks, but a fundamental understanding of how such network oscillators work remains elusive. Progress towards this goal has come from attempts to combine computational modeling approaches with conventional physiological analysis.


The cerebellum and VOR/OKR learning models. Although one particular model of the cerebellum, as proposed by Marr and Albus, provides a formal framework for understanding how heterosynaptic plasticity of Purkinje cells might be used for motor learning, the physiological details remain largely an engima. Developments in computational neuroscience and artificial neural networks applied to real control problems are essential to understand fully how workspace errors associated with movement performances can be converted into motor-command errors, and how these errors can then be used as one kind of synaptic input by motor-learning algorithms that are based on biologically plausible rules involving heterosynaptic plasticity. These developments, as well as recent advances in the study of cellular mechanisms of synaptic plasticity, form the basis for the detailed computational models of cerebellar motor learning that have been proposed. These models provide hints toward resolving a long-standing controversy in the oculomotor literature regarding the sites of adaptive changes in the vestibuloocular reflex (VOR) and the optokinetic eye movement response (OKR), and suggest new experiments to elucidate general mechanisms of sensory motor learning.


Rallpacks: a set of benchmarks for neuronal simulators. The field of computational neurobiology has advanced to the point where there are several general-purpose simulators to choose from. These cater to various niches in the world of realistic neuronal models, which range from the molecular level to descriptions of entire sensory modalities. In addition, there are numerous custom-designed simulations, adaptations of electrical circuit simulators, and other specific implementations of neurobiological models. As a first step towards evaluating this disparate set of simulators and simulations, and towards establishing standards for comparisons of speed and accuracy, we describe a set of benchmarks. These have been given the name 'Rallpacks' in honor of Wilfrid Rall, who pioneered the study of neuronal systems through analytical and numerical techniques.


Experimentalists and modelers: can we all just get along? How can the interaction between theoretical neuroscientists and their experimental counterparts be improved? This article discusses a number of suggestions relating to the presentation of data in experimental studies. In particular, published data should account for the diversity of response properties encountered, rather than concentrating on the 'representative' response, as well as emphasizing the stochastic nature of neurons by routinely including raw, unprocessed data from individual trials, which show the degree of variability prior to averaging.


[Immunoglobin-binding proteins in human placenta]. Phenomena of the binding of poor-soluble placenta proteins (PSPP) with pregnant women sera IgG as well as placenta blood IgG were studied. PSPP were extracted from the placenta tissue, washed out from soluble proteins, by the use of 3M KCl solution containing 0.005 M PMSF. PSPP were separated by the use of two-dimensional isoelectrofocusing and SDS-PAG electrophoresis and more than 30 different polypeptides were visualized. Having used various ELISA procedures with pregnant women sera IgG, placenta blood IgG as well as its Fab and Fc-fragments we have shown that both the receptor-type and the antigen-antibody-like interaction of PSPP took place. Both the polypeptide compositions and the isoelectrofocusing points ranges of the antigen-antibody-like interacting IgG-binding PSPP were determined by the use of the peroxidase conjugated Fab-fragments of the placenta blood IgG.


[Escherichia coli ribosomes having peptidyl-tRNA and deacylated tRNA at the A- and P-sites, respectively, may not be competent in translocation]. Ribosomes can have two states at 0 degree C: competent and noncompetent in translocation. In both states poly(U)-programmed ribosomes bind phenylalanyl-tRNA to A and P sites and form peptide bond. Elongation factor G promotes fast translocation in competent ribosomes and makes them noncompetent ones. Initial correlation between competent and noncompetent ribosomes is 2:1. Addition of deacylated tRNA does not influence phenomenon described as well as thermal reactivation of the ribosomes before beginning of the experiments. The possibility of deacylated tRNA translocation is shown. The translocation does not occurred provided that at least one of the ribosome sites is filled with shortened tRNA analog (tRNA with truncated CCA-end or tRNA anticodon arm).


[Imaging and functional parameters in diagnosis of obstructive nephropathy]. Obstruction of the kidney leads to terminal kidney failure within a few years. Therefore, early recognition of such obstruction is of importance. Non-invasive diagnostic ultrasound examination now allows intrauterine visualization of a suspected obstruction. However, the implications of such a dilated ureteral pelvic system are obscure. Whether there is obstruction or dilatation can only be evaluated postnatally by a nuclear technique. The aim of our study was to measure the recovery of kidney function. We investigated 13 kidneys of 9 newborns or small infants (up to 2 years). The follow-up was continuous for up to 29 days. The parameters were: urine output (24-h clearance), glomerular filtration rate, fractional excretion of sodium and potassium, free water clearance, total protein excretion, albumin and alpha 1 microglobulin excretion. The urine output fell from 0.3 to 0.12 ml/min within 14 days after relief of obstruction. The glomerular filtration rate rose from nearly 30 ml/min to about 50 ml/min within a week. The fractional excretion of sodium and potassium indicated recovery of the proximal tubli. The fractional sodium excretion fell below 1% within 4 days. The free water clearance reflects the concentrating ability of the kidney, and in kidneys from newborns it had only positive values, while in kidneys of children older than 6 months there were also negative values. The protein excretion and the albuminuria showed recovery of the glomerular as well as the tubular system.(ABSTRACT TRUNCATED AT 250 WORDS)


[Urethral stricture after TURP and transvesical prostatectomy]. In an attempt to elucidate factors predisposing to the occurrence of urethral stricture after transurethral resection of the prostate, we performed a prospective follow-up of 178 patients over 12-20 months. We took account of 11 factors that we considered important. Urethral strictures developed in 14.04% of the patients. The resection operations were carried out by five different surgeons, who had different rates of stricture. The only one of the 11 factors studied that was found to involve a statistically significant risk was the presence of an indwelling catheter for more than 3 days. No other factor influenced the result. This patient group was compared with a group of 73 patients followed up for 12-60 months following transvesical prostatectomy. In this group only one stricture (1.36% incidence rate) was observed retrospectively. It seems that urethral ischaemia might increase the risk of urethral stricture. Urethral injuries are considerably less frequent with open prostatectomy. Therefore, we recommend transvesical prostatectomy for pronounced prostatic hyperplasias.


[Effect of ecdysterone on the metabolism of purine and pyrimidine nucleotides in chicken tissues]. Hormonal amounts (10(-7) M) of ecdysterone phytoecdysteroid have been studied for their effect on the metabolic processes in the chicken tissues. Changes in the blood serum parameters which characterize the state of the integral systems of mineral, lipid, protein and nucleotide metabolism as well as the dynamics of metabolism of nucleic acids and their precursors--purine and pyrimidine nucleotides in the tissues with actively functioning protein-synthesizing system prove that the biological effect of ecdysterone after its introduction to the bird organism manifests both in the non-genome early phase of the action (0-1 h) and in the late genome phase (24-72 h), i.e. in analogy to the time of response to the introduction of classical steroid hormones. It is supposed that polyfunctional early effect of ecdysterone is mediated by its action on one of universal transmembrane signal systems of the cells--adenylate cyclase or polyphosphoinosite one, while the presence of the expressed changes in metabolism of nucleic acids in the late phase unambiguously indicate to its action in the bird organism as that of a typical steroid hormone.


[2 cases of anencephaly in one family].

[2 cases of anencephaly in one family]. Two births of anencephalic fetuses occurred in the second and the fourth pregnancies in a family. The children born after the 1st and 3rd pregnancies were normal. Anencephaly was not associated with spina bifida, and both fetuses were female. The study failed to demonstrate the cause of the abnormality.


The endemic treponematoses: not yet eradicated. The endemic treponematoses which comprise yaws, endemic syphilis (bejel) and pinta constitute a group of potentially disabling and disfiguring infections which primarily afflict children in tropical and subtropical areas. Foci where these diseases are now endemic have a patchy distribution and are typically confined to underprivileged communities living in remote rural areas, with little or no access to health services and removed from the mainstream of socioeconomic development. A drastic decline in the prevalence of these infections was brought about by the implementation of mass treatment campaigns with penicillin under the technical guidance of WHO and with material support from UNICEF in the 1950s and 1960s. These worldwide campaigns against the endemic treponematoses halted disease transmission in many areas and held the promise of complete eradication if intensive surveillance could be continued for some time with the increasing involvement of the basic health services. National campaigns were so successful that relatively low priority was given to the preparation of the rural health services for this new task. The failure of many countries to integrate active control measures into the functions of the rural health services led to the gradual build-up and extension of treponemal reservoirs and the resurgence of foci of increased disease transmission particularly in communities where standards of hygiene and health care had remained low. In a number of former endemic foci only low-level transmission persisted; in a few areas disease prevalence increased dramatically to reach pre-campaign levels. The lack of technical and financial resources limited the success of renewed national control activities in the most affected areas. Today, with waning interest in these diseases confined to remote, and thus silent, population groups, and a decreasing ability of health staff to identify cases, data collected by countries need to be supplemented by information from other sources in order to arrive at a more valid assessment of the situation concerning the endemic treponematoses. Central and West Africa are most severely affected by the resurgence of the endemic treponematoses. In recent years a number of countries (e.g. Ghana, Côte d'Ivoire and Mali) have launched renewed control efforts, often combining yaws or endemic syphilis control with other public health programmes. In Central Africa itinerant pygmy groups are still highly affected by yaws and are an important source of infection for the sedentary population with which they come into contact. In Chad, Sudan and Ethiopia, there is some evidence of persistent foci of endemic treponematoses; the epidemiological situation in Southern Africa is not well established.(ABSTRACT TRUNCATED AT 400 WORDS)


World malaria situation 1990. Division of Control of Tropical Diseases. World Health Organization, Geneva. Malaria risk of varying degree exists in 99 countries or areas. However, falciparum malaria does not exist or its relative prevalence is less than 1% in 13 of these countries. Accurate information on the global incidence of malaria is difficult to obtain because reporting is particularly incomplete in areas known to be highly endemic. The global incidence of malaria is estimated to be nearly 120 million clinical cases each year, with nearly 300 million people carrying the parasite. 90% of the total number of cases reported annually to WHO are from 19 countries only. This does not include the WHO African Region where reporting of cases remains fragmentary and irregular despite improvements in recent years. Some 75% of cases are concentrated in 9 countries (in decreasing order): India, Brazil, Afghanistan, Sri Lanka, Thailand, Indonesia, Viet Nam, Cambodia and China. Furthermore, within these countries malaria is concentrated in certain areas. Of a total world population of about 5.3 billion people, 3.1 billion (59%) live in areas free of malaria (it never existed, disappeared or was eliminated by antimalaria campaigns and the malaria-free status has been maintained). 1.7 billion people (32%) live in areas where endemic malaria was considerably reduced or even eliminated but transmission was reinstated and the situation is unstable or deteriorating. These latter areas include zones with the most severe malaria problems which developed following major ecological or social changes, such as agricultural or other economic exploitation of jungle areas, sociopolitical unrest, etc.; these zones comprise only about 1% of the world population. Areas where endemic malaria remains basically unchanged, and no national antimalaria programme was ever implemented, are inhabited by 500 million people (9%), mainly in tropical Africa. Severe malaria and mortality are caused by Plasmodium falciparum which is the predominant species of malaria in tropical Africa. In the rest of the world it is far less common. WHO receives very limited and irregular reports on malaria deaths. The vast majority of malaria deaths occur in Africa; estimates vary greatly: a figure of 800,000 deaths per year in African children has been quoted in 1991 by the WHO African Region. There are indications that mortality in children has fallen in some areas because of the widespread use of antimalarials, of social development and of better education. Countries in tropical Africa are estimated to have more than 80% of all clinical cases and more than 90% of all parasite carriers.(ABSTRACT TRUNCATED AT 400 WORDS)


Prevention of deep vein thrombosis in patients with hip fractures: low molecular weight heparin versus dextran. A randomized open trial was undertaken to compare the antithrombotic efficacy of a low molecular weight heparin (LMWH; Sandoparin) with that of dextran 70 in patients undergoing surgery for hip fracture. One hundred thirteen patients received LMWH once daily subcutaneously at a fixed dosage while 103 patients received intravenous dextran 70. Postoperative deep vein thrombosis (DVT) was assessed by a diagnostic algorithm using the 125Iodine fibrinogen uptake test as screening and Duplex ultrasonography and/or ascending venography as confirming techniques for suspected DVT. The frequency of DVT was significantly lower in the LMWH group than in the dextran group (15.5 versus 32.6%, p less than 0.005). Proximal DVT was rare in both groups (LMWH: 2%, Dextran: 1%). Only one case of fatal fat pulmonary embolism was observed during the 10 day prophylaxis period in a patient receiving Dextran. Three cases of pulmonary embolism occurred later; one fatal event in the dextran group on day 14, and two cases in the LMWH group (one fatal and one non-fatal event) on day 14 and 17, respectively. There was no major bleeding complication in either group. We conclude that the LMWH we used is safe, was well tolerated, and has a significantly better thromboprophylactic effect than dextran 70.


[In vitro effects of low molecular weight dextran on the surface properties of LDL]. The relative number of free amino groups on the surface of low density lipoproteins (LDL) was determined after the incubation with low molecular weight dextran (MW 40,000) in several concentrations by two different assays (fluorescamine, TNBS). A decrease of the detectable free amino groups of 40% was shown by both assays. After the incubation of lysine with dextran these changes could not be observed. These results are explained with the aggregation of LDL particles influenced by dextran.


Oligodendrocytes from optic nerves subjected to long term Wallerian degeneration retain the capacity to myelinate. It has previously been shown in the adult rat optic nerve that cells with many features of oligodendrocytes are capable of surviving for extended periods of time in the absence of axons. This is in contrast to the situation in the developing nervous system, where removal of axons leads to the failure of differentiation and to the death of oligodendrocytes. In the adult, these surviving oligodendrocytes were not typical in their appearance, and could only be identified with certainty using cell specific markers. In the present experiments, the functional capacity of these long-term quiescent cells to regenerate and myelinate was tested using the Shiverer mouse, a mutant lacking the gene for myelin basic protein (MBP), as a host animal. Fragments of optic nerve from adult rats which had been enucleated up to 2 years previously, were implanted into neonatal Shiverer mice. Four weeks later, the brains were removed and the formation of myelin investigated with antibodies to MBP, to ensure that this was of donor origin. Axons were demonstrated to have grown into the implants, and may have provided the stimulus for the production of MBP by the oligodendrocytes, which were stained positively within the implant. Myelin was demonstrated both within and adjacent to the implant. This study indicates that in the adult central nervous system, cells can survive for extended periods of time in the absence of axons, albeit in an inactive state, and are then capable of functional regeneration when placed in contact with unmyelinated axons. The origin of these cells, either from surviving oligodendrocytes which had previously myelinated the axons, or from progenitors lying within the adult nerve is unclear. The implications of these results are of importance in the further investigation of the potential for central nervous system regeneration.


[The production of autoantibodies to nerve tissue glycolipid antigens in patients with traumatic spinal cord injuries]. The production of autoantibodies (AAB) to the specific antigen of myelin-containing components of the nervous tissue galactocerebroside (GalC) was revealed in 117 patients with traumatic injuries to the spinal cord of various severity in periods of 18 months to 46 years after the injury. The degree of nervous system destruction, the type of the course of posttraumatic myelopathy, and the level of production of AAB to GalC were found interrelated. The existence of AAB provides evidence of sensitization of the immune system to nervous tissue antigens in spinal cord injury, while the level of AAB to GalC reflects the activity of the continuing destructive process due to the injuring effect of AAB under conditions of impaired permeability of the blood-brain barrier and shows the desirability of desensitization therapy.


Bradykinin stimulates Ca2+ entry via nitrendipine-sensitive Ca2+ channels in cultured human fibroblasts. Bradykinin stimulates Ca2+ entry in cultured human fibroblasts via nitrendipine-sensitive Ca2+ channels in cultured human fibroblasts. The Ca2+ entering via this pathway does not contribute directly to the elevation of cytosolic Ca2+, but rather appears to be specifically taken up into an internal Ca2+ pool.


[The dynamics of the quantitative changes in neurospecific proteins in the blood of neural cancer patients]. The authors studied the effect of a neurosurgical intervention on the dynamics of quantitative changes of neurospecific proteins (NSP(--alpha 1 and alpha 2 globulins of the brain (alpha GM1, alpha GM2), L2 glycoprotein (alpha GP2), and glyofibril acid protein (GFA)--in blood of neuro-oncological patients. High background NSP values were revealed in most of the 14 examined patients for the first time. A significant increase of the amount of proteins in the blood during anesthesia and in the course or the operation was found. Deviation of values from the average statistical parameters was encountered in patients with an unfavorable outcome. The results of the study point to marked changes of the blood-brain barrier in neuro-oncological patients.


Kallikrein-kinin system in the angiogenesis. Using the bioassay to investigate the presence of mediators in the angiogenesis exudate to explain its property to cause vascular permeability and fall in the blood pressure we found the presence of histamine, bradykinin, prostaglandin E2 and angiotensin in the exudate. However, the role of these mediators in the angiogenesis process needs to be investigated.


The epidermal growth factor precursor in the rat kidney seems to be processed by an aprotinin sensitive proteinase. Epidermal growth factor (EGF) is synthesized as a membrane bound precursor in the rat kidney. The precursor seems to be processed by an aprotinin sensitive proteinase. Intravenous infusion of aprotinin reduces the urinary excretion of EGF by 85% and increases the amount of renal EGF. Kidney membranes incubated at 37 degrees C release EGF and this release is inhibited by aprotinin.


Vasoactive effects of aprotinin.

Vasoactive effects of aprotinin. The protease inhibitor aprotinin was given a) in experimental septic shock, and b) in patients with hepatic cirrhosis and ascites, since in both conditions, activation of the plasma kallikrein-kinin system is associated with pathological systemic vasodilatation, which may trigger reflex neuroendocrine activation and renal solute retention. Given early in experimental sepsis, aprotinin maintained the arterial pressure, systemic vascular resistance (SVR), creatinine clearance and sodium excretion, all of which fell in controls. Aprotinin also blocked increases in pulmonary artery pressure and plasma renin activity (PRA). Given late in sepsis, aprotinin caused a rapid rise in arterial pressure and SVR towards baseline levels. In cirrhosis, aprotinin increased SVR in patients with low baseline values, and improved glomerular filtration rate, renal plasma flow and sodium excretion in all subjects; PRA was suppressed by aprotinin. Aprotinin reverses pathological systemic vasodilatation in these two conditions, and this is associated with a reduction in renin release and improved renal function.


Activation of the contact system in ascites from patients with gastrointestinal cancer. Our observations indicates that the plasma contact system is activated in ascites from patients with gastrointestinal cancer: Factor XII is activated, plasma kallikrein is present in complex with the protease inhibitor alpha 2-macroglobulin, and the plasma kallikrein substrate high molecular weight kininogen, is highly degraded. Contact activation seems to take place in spite of a high level of inhibition. Activation of the contact system generates mediators, which may play a role in the accumulation of ascites.


The effects of fractionated and unfractionated heparins with and without aprotinin on plasma inhibition of alpha and beta FX11a. Chromogenic peptide substrate assays were used to compare the effects of fractionated and unfractionated heparins on plasma inhibition of alpha and beta FX11a, with and without various concentrations of aprotinin. All of the heparins reduced beta FX11a inhibition at 1 or 2U/ml. Four heparins increased alpha FX11a inhibition. Aprotinin counteracted the reduction in beta FX11a inhibition and augmented the heparin potentiation of alpha FX11a inhibition.


Lack of significant unspecific effects of HOE 140 and other novel bradykinin antagonists in vitro and in vivo. The novel, potent and long-acting bradykinin (BK) antagonists, HOE 140, compound II and compound III, slightly decreased blood pressure, but did not affect heart rate and respiration of rats. The antagonists did not cause bronchoconstriction in guinea-pigs. Neither HOE 140 nor BK released histamine from isolated perfused hindlegs of rats. The lack of significant unspecific side effects of the novel antagonists of effective doses will further increase the usefulness of these compounds for experimental and therapeutic purposes.


Effects of bradykinin on ion conductances in NG108-15 neuroblastoma x glioma hybrid cells recorded with patch-clamp electrodes. Under whole-cell recording, bradykinin (BK) produced an initial outward membrane current followed by an inward current in voltage-clamped NG108-15 cells. The initial outward current was associated with a rise in intracellular Ca2+ and was accompanied by the opening of Ca(2+)-dependent K(+)-channels recorded with a cell-attached patch electrode. This current was inhibited by intracellular Mg2+. The inward current was associated with inhibition of the voltage-dependent K(+)-current IK(M). These effects accord with those previously observed in microelectrode-impaled cells, with the difference that BK produced much more pronounced and long-lasting desensitization in the patch-clamped cells.


Renal hyperfiltration states: relationship to kallikrein and kinins. Animal models and humans with glomerular hyperfiltration and hyperperfusion secondary to diabetes or high protein diet show increased renal production of kallikrein and kinins. Acute aminoacid infusion or ingestion also raises GFR, RPF and urinary kinins. Treatment with aprotinin or a kinin receptor antagonist reverses or prevents hyperfiltration in these rat models.


Stimulation of endothelial autacoid formation by inhibitors of angiotensin-converting enzyme. We have investigated in human endothelial cells in culture the effects of angiotensin-converting enzyme (ACE) inhibitors on the concentration of intracellular free Ca2+ ([Ca2+]i) and the formation of nitric oxide (NO) and prostacyclin (PGI2). Enalaprilat, moexiprilat and ramiprilat similarly potentiated the increase in [Ca2+]i elicited by bradykinin and caused an increase in resting [Ca2+]i when given alone. The latter effect was long-lasting and accompanied by an increased formation of NO and PGI2. All of these effects were inhibited by the B2-kinin receptor antagonist Hoe 140, suggesting that the endogenous synthesis/release of bradykinin represents an autocrine mechanism for the stimulation of endothelial autacoid formation. Thus these findings strongly support the concept that ACE inhibitors promote local vasodilation by increasing the level of bradykinin generated in subthreshold concentrations by the endothelium.


Essential role of kallikrein-kinin system in suppression of blood pressure rise during the developmental stage of hypertension induced by deoxycorticosterone acetate-salt in rats. Deoxycorticosterone acetate (DOCA)-salt hypertension was induced in Brown Norway (BN) kininogen-deficient rats (BN-Ka) and normal rats from the same strain (BN-Ki) after nephrectomy. Systolic blood pressure, which was determined by the tail-cuff method, of BN-Ki increased gradually during this treatment. In contrast, the blood pressure of mutant BN-Ka increased rapidly 2 weeks after the onset of the treatment. Urinary excretion of active kallikrein and prokallikrein increased at the same degree in rats of both strains during this treatment. Significant increase in urinary sodium excretion was observed with a tendency to increase in urine volume during the treatment in normal BN-Ki rats, whereas both parameters were essentially not increased in mutant BN-Ka rats, which could not generate urinary kinin. Aprotinin infusion by osmotic minipump to normal BN-Ki rats during the DOCA-salt treatment resulted in significant further increase in the systolic blood pressure.


Studies on components of the plasma kallikrein system as a risk factor in diabetics undergoing cardiopulmonary bypass. Components of the FX11-kallikrein systems were determined in blood samples from non diabetics and diabetics undergoing cardiopulmonary bypass (CPB). FX11 and prekallikrein levels fell in both groups with the largest falls in the diabetic group. Kallikrein inhibition was also lower in the diabetic group. Alpha-2-macroglobulin levels were lower in the diabetic group before operation and were markedly lower throughout CPB. Kallikrein like activities were also lower in the diabetic group. Beta FX11a inhibition values were higher in the diabetic group and fell in both groups during CPB. From the results obtained we concluded that the lower levels of FX11, prekallikrein, kallikrein inhibition and alpha-2-macroglobulin in the diabetic patients during CPB reflect enhanced activation of the FX11-plasma kallikrein systems. Blood loss in the diabetic group was higher than the for non diabetic group.