Neuronal death after trophic factor deprivation. Since the discovery of nerve growth factor (NGF) nearly 40 years ago, extensive studies have elaborated the trophic relationship between a neuron and its end organ. Much evidence has accumulated that depicts the crucial role of neurotrophic factors in determining neuronal survival both during development and after injury. The molecular events by which trophic factors determine whether an individual neuron lives or dies are poorly understood. The function of active protein synthesis as a prerequisite for neuronal death has led to the hypothesis that all neurons contain a mechanism to self-destruct or a phenomenon of programmed cell death. The loss of trophic support may, therefore, lead to the initiation of this so called "suicide" program within individual cells. Recently, multiple other trophic factors have been discovered. At least two of these factors, brain-derived neurotrophic factor and neurotrophin-3, appear related to members of the same gene family as the prototypic neurotrophic factor, nerve growth factor. Other metabolic factors, such as intracellular calcium, may be instrumental in determining the dependence of individual neurons on specific trophic support. A "set-point" hypothesis has been described concerning the relationship between intracellular calcium and the NGF dependency of embryonic neurons in cell culture. Changes in calcium metabolism may be an important determinant in altering trophic dependence and naturally occurring cell death. Neurotrophic factors also influence the reaction of the neuron to axotomy and subsequent axonal regeneration. Exogenously supplied trophic factors may prevent neuronal loss after axotomy when supplied at the site of injury. Both NGF and FGF have been shown to enhance early regeneration across gaps within silicone chambers.(ABSTRACT TRUNCATED AT 250 WORDS)


Uterotropic action in rats of amsonic acid and three of its synthetic precursors. Prompted by reports of sexual impotence among chemical factory workers exposed to amsonic acid (4,4'-diaminostilbene-2,2'-disulfonic acid; CAS 81-11-8) and its synthetic precursors 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNSDSA; CAS 128-42-7), 2-methyl-5-nitrobenzenesulfonic acid (MNBSA; CAS 121-03-09), and 4-nitrotoluene (CAS 99-99-0), the uterine-weight-increasing actions of single intraperitoneal doses of these chemicals were determined at 24 h after treatment in weanling female rats and compared to the results of similar experiments with diethylstilbestrol (DES; CAS 56-53-01), a synthetic estrogen chemically related to amsonic acid and DNSDSA. Doses of 100 mg/kg or less of amsonic acid were either without effects or produced equivocal effects, while uterine weights were increased after doses of 300 and 1000 mg/kg; doses of 3000 mg/kg were clearly toxic. Neither DNSDSA nor MNBSA increased uterine weight after doses that were not overtly toxic. Doses of 10 mg/kg or less of 4-nitrotoluene were without effect, but doses of 30 and 100 mg/kg increased uterine weights without producing overt toxicity; doses of 1000 mg/kg were clearly toxic. While both amsonic acid and 4-nitrotoluene exhibited uterotropic effects, they were both much weaker than DES in this respect. Other experiments indicated that the time course of the effects of approximately equiactive doses of amsonic acid and DES were very similar, and that the responses to oral doses of amsonic acid were not appreciably different from the responses to the same doses given intraperitoneally. Finally, a sample of amsonic acid taken from the workplace of the complaining workers was also found to have uterotropic activity. These experiments suggest that amsonic acid and 4-nitrotoluene have estrogenic activity, and thus provide a possible mechanistic explanation for the complaints of impotency in factory workers exposed to these substances.


[Current concept of the management of rheumatoid arthritis based on the pathogenesis]. A new therapeutic concept for the management of rheumatoid arthritis (RA) is strongly required since most traditional methods have failed to introduce remission. This paper attempted to review over novel therapeutic approaches based on the advanced program especially on early stage of RA. Clinical improvement has been observed with tripple combined therapy using such as bucillamine, methotraxate, and sufphasulzopyridine. However, more basic therapeutic program would be required based on the pathogenesis of RA. Here the future development of therapeutic approach including targetting immunosuppression or gene targetting on proliferated synovium has been discussed.


[Studies on Hanganutziu-Deicher antibodies in renal diseases]. H-D antibodies which were known as antibodies detectable in patients with serum sickness have recently been detected in renal diseases. We attempted to detect IgG, IgA and IgM antibodies to N-glycolyl GM3, among other H-D antibodies, by ELISA in various renal diseases and found increased IgM antibody in mesangial proliferative glomerulonephritis, membranoproliferative glomerulonephritis, IgA nephropathy, minimal change nephrotic syndrome and Henoch-Schönlein purpura nephritis (HSPN), elevated IgA antibody in IgA nephropathy and HSPN and raised IgG antibody in IgA nephropathy. In 2 cases of IgA nephropathy, there was noted a good correlation between clinical course and anti-N-glycolyl GM3 antibody titers. Measurement by ELISA of IgG antibody to cytomegalovirus (CMV) seemed as one of immune pathogenetic factors of IgA nephropathy showed a high positive rate for this antibody of IgA nephropathy patients and a positive correlation between the antibody and anti-N-glycolyl GM3 antibody. The key molecule of H-D antigens is N-glycolyl neuraminic acid (NGNA) and this sialic acid does not normally exist in humans. One can surmise, therefore, that in those renal diseases in which there was noted an elevation of anti-N-glycolyl GM3 antibody, this antigen is formed or generated by some unknown mechanism. In other words, it may be that humans are not entirely negative for H-D antigens but have a minimum inherent antigenicity and a potential capacity to synthesize these antigens.(ABSTRACT TRUNCATED AT 250 WORDS)


Computer simulation of neurone pattern processing. Physiological simulation techniques can provide a very useful additional tool with which to explore the behaviour of neurones, especially for probing those areas that are not easily accessible to experimental monitoring techniques such as activity in the dendrites. This paper explores the information processing capabilities of a cerebral cortical pyramidal neurone using a computer simulation. The computer model simulated two important aspects of pyramidal cell behaviour: the transient distribution of membrane potential over a proportion of the apical dendritic tree of the cell, and the ability of the receptors in the neurones' dendritic spines to undergo plastic changes. The simulation of receptor behaviour modelled the ability of receptor regions to increase their sensitivity if a prior input had occurred when the spine head membrane was depolarized. The model was used to explore the response of the cell to different inputs. It underlined the need for large amounts of local activity before spinal receptors changed their sensitivity. It also demonstrated the ability of pyramidal cells to both recognize patterns of input and to associate one pattern of inputs with another. It is argued that this pattern association property could be a neuronal basis for association learning.


Minimal residual disease in childhood acute lymphoblastic leukemia: analysis of patients in continuous complete remission or with consecutive relapse. Differences in tumor cell burden among acute lymphoblastic leukemia (ALL) patients are largely unexplored, because methods of detecting residual malignant cells have not been sufficiently sensitive. Using the polymerase chain reaction (PCR) amplification of rearranged T-cell receptor delta(TCR delta)-chain junctional sequences for the preparation of clonospecific probes, we performed a retrospective PCR study of remission bone marrow (BM) samples in seven pediatric patients with ALL who subsequently relapsed (the largest series studied so far) and in 10 patients who were in longterm (greater than 39 to greater than 72 months) remission. Following two rounds of PCR primed by nested amplimers, 1 x 10(-4) to 1 x 10(-6) cells could be identified in 16 out of 17 cases. PCR analysis of 39 BM and peripheral blood samples obtained from ALL patients considered to be in complete remission according to morphological criteria revealed the following results. In BM remission specimens of all 10 patients in continuous complete remission for a long time (median 55 months), no residual leukemic cells could be identified in the latest remission sample available for PCR analysis. In three patients the persistence of residual leukemic cells, or the continuous increase of residual blasts to the point of clinical manifestation, were indicative of impending relapse. In three patients PCR analysis failed to identify residual leukemic cells in BM samples obtained 2, 6 and 16 months respectively before clinical relapse. Differences in the duration of minimal residual disease were not associated with distinct clinical-hematological features. In one patient a different pattern of V delta 2 recombination occurred in leukemic cells from diagnosis to relapse, thus preventing the further monitoring of the patient by the initial clonospecific probe.


Depolarization- and agonist-regulated expression of neuronal metabotropic glutamate receptor 1 (mGluR1). In established 8-12-day-old primary cultures of differentiated rat cerebellar granule neurons the level of metabotropic glutamate receptor 1 (mGluR1) mRNA and the sensitivity of cultures to the agonist-stimulated inositol phosphate (IP) formation was reversibly modified by changing the depolarizing properties of the medium, i.e. the medium KCl concentration. The mGluR1 mRNA content was suppressed by increasing the medium KCl content and elevated by decreasing it. The mGluR agonist quisqualate inhibited the mGluR1 expression. This is the first direct demonstration of a differential expression of neuronal mGluR1 in an established neuronal culture. The model can be used to study the molecular mechanism of neuronal plasticity.


Inhibition by angiotensin converting enzyme inhibitors of endothelin secretion from cultured human endothelial cells. We conducted a study to determine whether angiotensin converting enzyme inhibitors (ACEIs) inhibit endothelin secretion from cultured human endothelial cells. Confluent umbilical vein endothelial cells were incubated in multi-well plates with culture medium containing either captopril (10(-6), 10(-5), 10(-4) M) or enalaprilat (10(-7), 10(-6), 10(-5) M) for 6 hours. Immunoreactive endothelin in the medium was measured by radioimmunoassay. Calf serum (CS) stimulated endothelin release in a concentration-dependent manner, and both ACEIs inhibited 5% CS-stimulated endothelin release in a concentration-dependent manner. To explore the mechanisms of ACEI-induced suppression of endothelin release, the effects of angiotensin II (10(-8), 10(-7), 10(-6) M), angiotensin converting enzyme (0.1, 1, 10 mU/ml), bradykinin (10(-8), 10(-7), 10(-6) M), and sodium nitroprusside (10(-6), 10(-5), 10(-4) M) on endothelin release were also examined. Although angiotensin II and angiotensin converting enzyme had no significant effect on endothelin release, concentration-dependent suppression occurred with bradykinin and sodium nitroprusside. These results indicate that ACEIs inhibit the stimulated release of endothelin from human endothelial cells, and provide indirect evidence that ACEI-induced ET suppression may be mediated via potentiation of autacoid formation from the cells.


Recent developments in the classification and functional significance of receptors for ATP and UTP, evidence for nucleotide receptors. The presence of a nucleotide receptor activated with similar potencies by UTP and ATP is suggested by recent data from a variety of different cell types. This receptor type appears distinct from previously described ATP-sensitive P2-purinoceptor subtypes and, probably, from other UTP-sensitive receptors, however further studies using selective antagonists are necessary to provide a definitive characterisation. Although the functional role of endogenous extracellular ATP has already achieved recognition there are also many diverse examples of cells and tissues which respond to UTP at micromolar or sub-micromolar concentrations. Therefore, the possible physiological importance of UTP is a fertile area for further investigation. The functional significance of ATP/UTP receptors is underlined by recent demonstrations that UTP and ATP modulate chloride ion secretion in human airways epithelium, possibly by activation of a nucleotide receptor, an effect which may have potential clinical utility in the treatment of cystic fibrosis.


Effects of pentobarbital tolerance and dependence on convulsant and GABAA receptor antagonist binding. Experiments were performed which examined the effects of pentobarbital tolerance and dependence on GABAA receptor antagonist binding. In rats implanted with pentobarbital pellets for 7 days, followed by 24 hours of withdrawal, there was a significant decrease in the latency of TBPS-induced seizures and an increase in [35S]TBPS binding in the frontal cortex. The pentobarbital tolerant rats had a significant increase in the low affinity KD of [3H]SR95531 binding. Removal of the pellets for 24 hours caused a reversal of the effect on the low affinity KD and caused a decrease in the number of low affinity binding sites. In vitro addition of pentobarbital to binding assays produced a decrease in the number of high affinity [3H]SR95531 binding sites without changing low affinity binding. In the cerebellum, the binding in none of the treatment groups was significantly different from placebo. These observations suggest that pentobarbital tolerance and withdrawal cause changes in the properties of the GABAA receptor antagonist binding site which are different from those caused by in vitro exposure to the drug.


2[125I]iodomelatonin binding sites in spleens of guinea pigs. 2-[125I]Iodomelatonin was found to bind specifically to the membrane preparations of the spleens of guinea pigs with high affinity. The binding was rapid, stable, saturable and reversible. Scatchard analysis of the binding assays revealed an equilibrium dissociation constant (Kd) of 49.8 +/- 4.12 pmol/l and binding site density (Bmax) of 0.69 +/- 0.082 fmol/mg protein at mid-light (n = 10). There was no significant change in the Kd (41.8 +/- 3.16 pmol/l) or the Bmax (0.58 +/- 0.070 fmol/mg protein) at mid-dark (n = 10). Kinetic analysis showed a Kd of 23.13 +/- 4.81 pmol/l (mean +/- SE, n = 4), in agreement to that derived from the saturation studies. The 2-[125I]iodomelatonin binding sites have the following order of potency: 2-iodomelatonin greater than melatonin greater than 6-chloromelatonin much greater than N-acetylserotonin, 6-hydroxymelatonin greater than 5-methoxytryptamine, 5 methoxytryptophol greater than serotonin, 5-methoxyindole-3-acetic acid greater than 5-hydroxytryptophol, 3-acetylindole, 1-acetylindole-3-carboxyaldehyde, L-tryptophan greater than tryptamine, 5-hydroxyindole-3-acetic acid. Differential centrifugation studies showed that the binding sites are localized mainly in the nuclear fraction (65.5%), the rest are distributed in the microsomal fraction (17.4%), mitochondrial fraction (14.7%) and cytosolic fraction (0.3%). The demonstration of 2-[125I]iodomelatonin binding sites in the spleen suggests the presence of melatonin receptors and a direct mechanism of action of melatonin on the immune system.


Organization and function of the mcrBC genes of Escherichia coli K-12. Many natural DNA sequences are restricted in Escherichia coli K-12, not only by the classic Type I restriction system EcoK, but also by one of three modification-specific restriction systems found in K-12. The McrBC system is the best studied of these. We infer from the base composition of the mcrBC genes that they were imported from an evolutionarily distant source. The genes are located in a hypervariable cluster of restriction genes that may play a significant role in generation of species identity in enteric bacteria. Restriction activity requires the products of two genes for activity both in vivo and in vitro. The mcrB gene elaborates two protein products, only one of which is required for activity in vitro, but both of which contain a conserved amino acid sequence motif identified as a possible GTP-binding site. The mcrC gene product contains a leucine heptad repeat that could play a role in protein-protein interactions. McrBC activity in vivo and in vitro depends on the presence of modified cytosine in a specific sequence context; three different modifications are recognized. The in vitro activity of this novel multi-subunit restriction enzyme displays an absolute requirement for GTP as a cofactor.


Characterization of cat messenger RNA decay suggests that turnover occurs by endonucleolytic cleavage in a 3' to 5' direction. Turnover of the chloramphenicol acetyltransferase (cat) messenger RNA in Escherichia coli was investigated by analysis of cellular mRNA decay intermediates and the transcript sequence. Analysis of the sequence has revealed the presence of a repetitive extragenic palindromic (REP) element at the 3' end of the transcript as well as several 5'-UUAU-3' sequences, two elements which have roles in modulating turnover of other E. coli mRNAs. For cat mRNA, however, removal of the REP sequence has no effect on the half-life of the transcript, indicating that the REP sequence does not stabilize the upstream region of this message. Results from mapping of the mRNA decay products by several techniques suggest that the message is instead subject to endonucleolytic attack at five sites 5' of the REP element. The sequence UUAU is present at three of these five sites. It also appears that the cat mRNA is sequentially cleaved in a 3' to 5' direction during turnover of this mRNA in vivo. A model for cat mRNA turnover is discussed.


Biochemical characterization of Escherichia coli DNA helicase I. The gene product of F tral is a bifunctional protein which nicks and unwinds the F plasmid during conjugal DNA transfer. Further biochemical characterization of the Tral protein reveals that it has a second, much lower, Km for ATP hydrolysis, in addition to that previously identified. Measurement of the single-stranded DNA-stimulated ATPase rate indicates that there is co-operative interaction between the enzyme monomers for maximal activity. Furthermore, 18O-exchange experiments indicate that Tral protein hydrolyses ATP with, at most, a low-level reversal of the hydrolytic step during each turnover.


Ca2+ potentiates corticotropin-induced, but not isoproterenol-induced, [3H]guanosine diphosphate release in rat adipocyte membranes. EGTA abolished corticotropin (ACTH)-stimulated adenylate cyclase in rat adipocyte membranes. In contrast, the potency of guanosine triphosphate (GTP) stimulation of adenylate cyclase activated with ACTH was greater in the presence of Ca2+ (1 mmol/L). EGTA (1 mmol/L) powerfully inhibited ACTH-stimulated [3H]guanosine diphosphate (GDP) release from membranes prelabeled with [3H]GTP in the presence of isoproterenol (ISO) or ACTH, whereas Ca2+ significantly increased it. In contrast, neither EGTA nor Ca2+ affected ISO-stimulated [3H]GDP release. These data clearly show that Ca2+ is necessary for the binding of ACTH to its receptor, and that Ca2+ stimulates the interaction of the ACTH-occupied receptor with GTP-binding proteins.


Growth factor receptor regulation in the Minn-1 leprechaun: defects in both insulin receptor and epidermal growth factor receptor gene expression. Leprechaunism is a disorder characterized by intrauterine growth retardation, distinctive dysmorphology, and extreme insulin resistance due to structural abnormalities of the insulin receptor (IR). In addition to the IR, it has been suggested that abnormalities of the other growth factor receptors may occur in this syndrome. Using fibroblasts from the Minn-1 leprechaun, we have now investigated the expression of three different growth factor receptor genes: the IR, the insulin-like growth factor-I receptor (IGF-IR), and the epidermal growth factor receptor (EGFR). In agreement with previous studies, we found decreased insulin binding to fibroblasts from the Minn-1 leprechaun. In these cells, the IR transcription rate was not decreased, and sequence analysis of the IR promoter region of the patient showed no abnormalities. Both single-stranded conformational polymorphism analysis (SSCP) and DNA sequencing confirmed a previously reported nonsense mutation in one of the patient's two IR alleles at exon 14. mRNA levels for the IR were markedly decreased, suggesting that IR mRNA turnover was enhanced. We then studied the expression of the closely related IGF-IR Ligand binding, mRNA content, and transcription rate were all normal. In contrast to the IGF-IR, when the EGFR was studied, ligand binding and mRNA content were markedly decreased. These studies therefore raise the possibility that the phenotypic expression of leprechaunism results from defects in the expression of both the IR and the EGFR.


[Antibody levels against coxsackie B viruses in patients with type 1 diabetes mellitus]. In order to detect if the presence of interrelationship between Coxsackie B Viruses (CBV) and etiology of type 1 diabetes mellitus, antibodies to CBV by microneutralization test serotypes 1, 2, 3, 4, 5 and 6 were investigated in 37 sera obtained from patients with type 1 diabetes mellitus and 100 sera from healthy individuals as control group. No significant difference could found that neutralizing antibody levels to CBV serotypes between in sera obtained from patient with type 1 diabetes mellitus and sera obtained from control group. As a result, interrelationship was not detected between CBV infection and having type 1 diabetes mellitus.


Effects of professional and media warnings about the association between aspirin use in children and Reye's syndrome. The media played a central role in changing the use of aspirin among children with viral illness following reports of its association with a rare but deadly disease, Reye's syndrome (RS). It did so by alerting health professionals and parents about ways to prevent RS in children. Indeed, by the time aspirin product labeling was required by the FDA in 1986, most of the decline in RS incidence had already occurred. In the past, media-only health education campaigns have been relatively unsuccessful in achieving long-term changes in complex health behaviors. This article supports the theory that media warnings about the hazards of common products may successfully change consumer behaviors when the illness is devastating, the behavioral message is simple, acceptable and inexpensive alternatives are available, and the campaign is comprehensive, involving multiple professional groups to reinforce direct appeals to consumers.


HIV-infected health care professionals: public threat or public sacrifice? The ethical controversy surrounding the Centers for Disease Control (CDC) and American Medical Association (AMA) guidelines for restricting the practice of HIV-infected health professionals appears to hinge on whether we give priority to the rights of infected workers or patients. We cannot simply dismiss the concerns of patients as irrational, despite the low risks of transmission. Nor can we avoid the dispute about rights by claiming with the AMA that professionals have obligations to refrain from imposing "identifiable risks," however low, on patients. Nevertheless, allowing the full exercise of patient rights, either by giving patients the opportunity to know the risks they face and to switch providers, or by removing infected providers (compulsory switching), would make each of us worse off. This gives us adequate reason to reject these guidelines and to emphasize other infection control measures.


Risky business: setting public health policy for HIV-infected health care professionals. An analysis of the restrictive proposals provoked by the case of Kimberly Bergalis and four other patients apparently infected with HIV during the course of dental treatment reveals that they resulted from an inability to evaluate appropriately the infinitesimal risk of HIV transmission from practitioner to patient. The proposals also resulted from an effort to create risk prevention policy without appreciating the distinction between regulating things or procedures, which have no human rights, and regulating people, who have rights that should not be infringed without serious justification. This analysis demonstrates that the proposed restrictive policies are not justified because they do nothing to prevent the spread of HIV, and they cause unnecessary and substantial harm to health care practitioners.


Accumbens-caudate-septal circuit as a system for hippocampal regulation: involvement of a GABAergic neurotransmission. Hippocampal-based epileptiform activity may reach the basal ganglia via the nucleus accumbens. Previous data suggested that caudate nucleus is able to influence hippocampal epilepsy, probably sending a projection to the septum. In order to test the hypothesis of a retrograde activation of accumbens-caudate pathway in hippocampal regulation, we electrically stimulated both caudate nucleus and nucleus accumbens and studied modifications of hippocampal EEG in the feline focal epilepsy model. We also performed bilateral electrolytic lesion of nucleus accumbens and repeated caudate stimulation. Results showed that nucleus accumbens stimulation was ineffective in modifying hippocampal epilepsy; on the contrary, caudate stimulation caused a statistically significant decrease of hippocampal spike frequency and amplitude. On the other hand, in accumbens-lesioned animals caudate activation consistently reduced hippocampal epilepsy to a significant degree. As the caudate nucleus influences hippocampal activity and the septum may constitute a relay station of this functional relation, a possibility was tested concerning a GABAergic mediation. To this end, after a stable caudate-induced effect was reached, an intraseptal microinjection of picrotoxin (GABA receptor antagonist) was made and caudate stimulation repeated at the same parameters. Such a study showed that after intraseptal picrotoxin, caudate stimulation failed to elicit any type of modification of hippocampal activity. Experimental findings support the notion that the striatal modulation on hippocampus is mediated by an anterograde rather than a retrograde pathway, and underline the possibility of a GABAergic caudate-septal influence.


Prevention and control of influenza. Recommendations of the Immunization Practices Advisory Committee (ACIP). These recommendations update information on the vaccine and antiviral agents available for controlling influenza during the 1992-1993 influenza season (superseding the MMWR 1991; 40(no. RR-6): 1-15.) The primary changes include statements about vaccination of persons with known hypersensitivity to eggs or other components of the influenza vaccine, the optimal timing of influenza vaccination, and the influenza strains in the trivalent vaccine for 1992-1993.


Cold-sensitive mutants of p34cdc2 that suppress a mitotic catastrophe phenotype in fission yeast. The p34cdc2 protein kinase plays a central role in the regulation of the eukaryotic cell cycle, being required both in late G1 for the commitment to S-phase and in late G2 for the initiation of mitosis. p34cdc2 also determines the precise timing of entry into mitosis in fission yeast, where a number of gene products that regulate p34cdc2 activity have been identified and characterised. To investigate further the mitotic role of p34cdc2 in this organism we have isolated new cold-sensitive p34cdc2 mutants. These are defective only in their G2 function and are extragenic suppressors of the lethal premature entry into mitosis brought about by mutating the mitotic inhibitor p107wee1 and overproducing the mitotic activator p80cdc25. One of the mutant proteins p34cdc2-E8 is only functional in the absence of p107wee1, and all the mutant strains have reduced histone H1 kinase activity in vitro. Each mutant allele has been cloned and sequenced, and the lesions responsible for the cold-sensitive phenotypes identified. All the mutations were found to map to regions that are conserved between the fission yeast p34cdc2 and functional homologues from higher eukaryotes.


The Streptomyces coelicolor whiB gene encodes a small transcription factor-like protein dispensable for growth but essential for sporulation. A non-sporulating mutant (whiB218) of Streptomyces coelicolor A3(2), which proved to contain a deletion of more than 5 kb of DNA including whiB, was complemented by a small cloned DNA fragment, deduced from DNA sequencing to encode a protein of only 87 amino acids. This protein would bear some similarities to transcription factors, including an acidic, somewhat amphipathic alpha-helical region predicted near its N-terminus, and a basic alpha-helical region predicted at its C-terminus. A point mutation (whiB70) giving a phenotype indistinguishable from that of the whiB218 deletion mutant would cause a leucine to proline change at the start of the latter region. The whiB homologue from the closely related species S. lividans differed at only one base from its S. coelicolor counterpart, and would specify an identical polypeptide.


Expression of yeast cytochrome c1 is controlled at the transcriptional level by glucose, oxygen and haem. The nuclear gene for cytochrome c1 in Saccharomyces cerevisiae (CYT1) was localized on chromosome XV. Its upstream region was identified by functional complementation. Fusion to the lacZ reporter gene on a CEN plasmid allowed study of the effect of carbon sources and of specific deletion mutations on expression of the gene in yeast transformants. Detailed promoter analysis combined with expression studies in recipient strains defective in regulatory genes identified cis-acting sites and transcription factors involved in the regulated expression of the cytochrome c1 gene. These analyses showed that, in the presence of glucose, transcription of CYT1 is positively controlled by oxygen, presumably through the haem signal, and mediated by the HAP1-encoded transactivator. It is additionally regulated by the HAP2/3/4 complex which mediates gene activation mainly under glucose-free conditions. Basal transcription is, in part, effected by CPF1, a centromere and promoter-binding factor.


Neuropeptide Y1 subtype pharmacology of a recombinantly expressed neuropeptide receptor. Neuropeptide Y (NPY) is an important central and peripheral modulator of neural and endocrine functions. This neuropeptide interacts with at least two pharmacologically distinct receptors, termed Y1 and Y2. At Y1 receptors, the NPY analog [Leu31,Pro34] NPY, but not the carboxyl-terminal fragment NPY-(18-36), displaces radiolabeled NPY and the sequence-related peptide YY, whereas Y2 receptors exhibit the opposite selectivity. We have used cultured mammalian 293 cells for the high level transient expression of a previously cloned putative neuropeptide receptor of rat brain. We report that this receptor displays the ligand binding properties and selectivity of a Y1 receptor, with a single high affinity site for 125I-NPY (Kd, 0.7 +/- 0.2 nM). The functionality of the recombinantly expressed receptor was demonstrated by an inhibition of adenylyl cyclase and a concomitant mobilization of intracellular Ca2+.


Identification of three separate guanine nucleotide-binding proteins that interact with the delta-opioid receptor in NG108-15 neuroblastoma x glioma hybrid cells. Five separate guanine nucleotide-binding proteins (G proteins) were immunologically identified in membranes from neuroblastoma x glioma NG108-15 hybrid cells. These alpha subunit proteins were Gi2 alpha, two isoforms of Gi3 alpha, and two isoforms of Go alpha. The G proteins that interacted with delta-opioid receptors in these membranes were identified using cholera toxin (CTX)-induced ADP-ribosylation and antisera selective for various G protein alpha subunits. In the presence of delta-opioid agonists, CTX induced the incorporation of [32P]ADP-ribose into three pertussis toxin substrates. Using antisera generated against peptide sequences from G alpha subunits, these three pertussis toxin substrates were identified as Gi2 alpha, Go2 alpha, and one isoform of Gi3 alpha, which has yet to be identified. This CTX-induced labeling was demonstrated to be mediated via the delta-opioid receptor in these hybrid cells by the observation that delta agonists D-Ala2-D-Leu5-enkephalin (DA-DLE) and D-Pen2-D-Pen5-enkephalin, as well as the nonselective agonists etorphine and bremazocine, were active, but the mu agonist PL017 and the kappa agonist U-50-488H did not show this activity. This incorporation into all three substrates induced by DADLE was dose dependent, with EC50 (95% confidence interval) values ranging from 12 (3-52) to 183 (65-520) nM, which compared with the Kd value of 10 +/- 1.5 nM for this agonist, a dose that produces maximal inhibition of adenylate cyclase activity. Furthermore, pretreatment of the cells with pertussis toxin or treatment of the membranes with the antagonist naloxone blocked the incorporation induced by DADLE. Incorporation of [32P]ADP-ribose into all three substrates decreased 35-83% in membranes in which the receptors had been down-regulated by chronic treatment of the cells with DADLE. Thus, a single opioid receptor type can interact with three separate G proteins.


Differences in effects of forskolin and an analog on calcium currents in cardiac myocytes suggest intra- and extracellular sites of action. The effects of forskolin (FO) and a water-soluble derivative of FO, L858051 (7 beta-desacetyl-7 beta-[gamma-(N-methylpiperazino)-butyryl] forskolin), were compared on calcium currents (ICa) studied by the whole-cell patch-clamp technique in frog ventricular cardiac myocytes. Both FO and L858051 increased ICa, with half-times of 160 +/- 20 sec and 343 +/- 22 sec, respectively. The stimulation was blocked by internal perfusion with inhibitors of protein kinase A. The EC50 for stimulation of ICa was 0.3 microM for FO and 1.0 microM for L858051. The maximal stimulated current was the same for both drugs, 20.3 microA/cm2 and 23.1 microA/cm2, respectively. Internal perfusion with 30-500 microM guanylyl 5'-imidodiphosphate [Gpp(NH)p] suppressed ICa stimulation by low concentrations of FO or L858051. This suppression was due to a rightward shift in the concentration-response curve, with increases in the EC50 values to 11.4 microM for FO and 28.4 microM for L858051. Isoproterenol (ISO) was ineffective in increasing ICa after the FO-stimulated ICa had been reduced by Gpp(NH)p and FO had been washed out. In contrast, after the L858051-stimulated current had been reduced by Gpp(NH)p, ISO stimulated ICa significantly. This stimulation was blocked by inhibitors of protein kinase A and was due to a positive effect of L858051 not shared by FO. A brief application of L858051 after Gpp(NH)p had blocked the ISO response restored the ISO response for at least 30 min. This effect was mimicked by internal perfusion with low concentrations of L858051. We conclude that the ability of brief exposure of L858051, but not FO, to restore the response to ISO after Gpp(NH)p is due to the accumulation of L858051 intracellularly, due to its hydrophilicity. Because internal L858051 and FO are very ineffective in stimulating adenylyl cyclase, whereas internal L858051 can restore the ISO response blocked by Gpp(NH)p, we propose that FO compounds can affect adenylyl cyclase at two sites, one site that is accessible only from the extracellular side that stimulates catalytic activity and another that is accessible from the intracellular side that increases beta-agonist efficacy in the presence of Gpp(NH)p.


Free radical metabolism of halothane in vivo: radical adducts detected in bile. Two radical adduct species have been detected in the bile of living rats treated with halothane and phenyl-N-t-butylnitrone (PBN). The treatment of rats with 12% oxygen was required for radical adduct detection. Analysis of the corresponding EPR spectra obtained when deuterated PBN and deuterated halothane or [2-13C]halothane was used shows that these two species result from the spin trapping of two halothane-derived free radicals. Coupling constants were aN = 15.72 G, a beta H = 2.09 G, a gamma H = 0.79 G, and aF = 0.63 G(3F) and aN = 15.16 G, a beta H = 4.14 G, a gamma H = 0.48 G, and aF = 0.3 G(3F) for the two species. Two radical adducts with similar coupling constants were detected when halothane was reduced by zinc dust in the presence of PBN, suggesting that the formation of these two distinct species from halothane can be attributed to the one-electron reduction of halothane and the formation of diastereomeric radical adducts. The identification of both radical adducts as halothane-derived species indicates that there is no in vivo EPR evidence for lipid radical formation during halothane intoxication, as had previously been reported.


Singlet oxygen: a potential culprit in myocardial injury? The purpose of this study was to explore the role of singlet oxygen in cardiovascular injury. To accomplish this objective, we investigated the effect of singlet oxygen [generated from photoactivation of rose-bengal] on the calcium transport and Ca(2+)-ATPase activity of cardiac sarcoplasmic reticulum and compared these results with those obtained by superoxide radical, hydrogen peroxide and hydroxyl radical. Isolated cardiac SR exposed to rose bengal (10 nM) irradiated at (560 nm) produced a significant inhibition of Ca2+ uptake; from 2.27 +/- 0.05 to 0.62 +/- 0.05 mumol Ca2+/mg.min (mean +/- SE) (P less than 0.01) and Ca(2+)-ATPase activity from 2.08 +/- 0.05 mumol Pi/min.mg to 0.28 +/- 0.04 mumol Pi/min.mg (mean +/- SE) (P less than 0.01). The inhibition of calcium uptake and Ca(2+)-ATPase activity by rose bengal derived activated oxygen (singlet oxygen) was dependent on the duration of exposure and intensity of light. The singlet oxygen scavengers ascorbic acid and histidine significantly protected SR Ca(2+)-ATPase against rose bengal derived activated oxygen species but superoxide dismutase and catalase did not attenuate the inhibition. SDS-polyacrylamide gel electrophoresis of SR exposed to photoactivated rose bengal up to 14 min, demonstrated complete loss of Ca(2+)-ATPase monomer band which was significantly protected by histidine. Irradiation of rose bengal also caused an 18% loss of total sulfhydryl groups of SR. On the other hand, superoxide (generated from xanthine oxidase action on xanthine) and hydroxyl radical (0.5 mM H2O2 + Fe(2+)-EDTA) as well as H2O2 (12 mM) were without any effect on the 97,000 dalton Ca(2+)-ATPase band of sarcoplasmic reticulum.(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of oxygen free radicals generated during vanadate-stimulated NADH oxidation. The oxidation of NADH and accompanying reduction of oxygen to H2O2 stimulated by polyvanadate was markedly inhibited by SOD and cytochrome c. The presence of decavanadate, the polymeric form, is necessary for obtaining the microsomal enzyme-catalyzed activity. The accompanying activity of reduction of cytochrome c was found to be SOD-insensitive and therefore does not represent superoxide formation. The reduction of cytochrome c by vanadyl sulfate was also SOD-insensitive. In the presence of H2O2, all the forms of vanadate were able to oxidize reduced cytochrome c, which was sensitive to mannitol, tris and also catalase, indicating H2O2-dependent generation of hydroxyl radicals. Using ESR and spin trapping technique only hydroxyl radicals, but not superoxide anion radicals, were detected during polyvanadate-dependent NADH oxidation.


Specific transcription factors stimulate simian virus 40 and polyomavirus origins of DNA replication. The origins of DNA replication (ori) in simian virus 40 (SV40) and polyomavirus (Py) contain an auxiliary component (aux-2) composed of multiple transcription factor binding sites. To determine whether this component stimulated replication by binding specific transcription factors, aux-2 was replaced by synthetic oligonucleotides that bound a single transcription factor. Sp1 and T-antigen (T-ag) sites, which exist in the natural SV40 aux-2 sequence, provided approximately 75 and approximately 20%, respectively, of aux-2 activity when transfected into monkey cells. In cell extracts, only T-ag sites were active. AP1 binding sites could replace completely either SV40 or Py aux-2. Mutations that eliminated AP1 binding also eliminated AP1 stimulation of replication. Yeast GAL4 binding sites that strongly stimulated transcription in the presence of GAL4 proteins failed to stimulate SV40 DNA replication, although they did partially replace Py aux-2. Stimulation required the presence of proteins consisting of the GAL4 DNA binding domain fused to specific activation domains such as VP16 or c-Jun. These data demonstrate a clear role for transcription factors with specific activation domains in activating both SV40 and Py ori. However, no correlation was observed between the ability of specific proteins to stimulate promoter activity and their ability to stimulate origin activity. We propose that only transcription factors whose specific activation domains can interact with the T-ag initiation complex can stimulate SV40 and Py ori-core activity.


The 19-kilodalton adenovirus E1B transforming protein inhibits programmed cell death and prevents cytolysis by tumor necrosis factor alpha. The adenovirus E1A and E1B proteins are required for transformation of primary rodent cells. When expressed in the absence of the 19,000-dalton (19K) E1B protein, however, the E1A proteins are acutely cytotoxic and induce host cell chromosomal DNA fragmentation and cytolysis, analogous to cells undergoing programmed cell death (apoptosis). E1A alone can efficiently initiate the formation of foci which subsequently undergo abortive transformation whereby stimulation of cell growth is counteracted by continual cell death. Cell lines with an immortalized growth potential eventually arise with low frequency. Coexpression of the E1B 19K protein with E1A is sufficient to overcome abortive transformation to produce high-frequency transformation. Like E1A, the tumoricidal cytokine tumor necrosis factor alpha (TNF-alpha) evokes a programmed cell death response in many tumor cell lines by inducing DNA fragmentation and cytolysis. Expression of the E1B 19K protein by viral infection, by transient expression, or in transformed cells completely and specifically blocks this TNF-alpha-induced DNA fragmentation and cell death. Cosegregation of 19K protein transforming activity with protection from TNF-alpha-mediated cytolysis demonstrates that both activities are likely the consequence of the same function of the protein. Therefore, we propose that by suppressing an intrinsic cell death mechanism activated by TNF-alpha or E1A, the E1B 19K protein enhances the transforming activity of E1A and enables adenovirus to evade TNF-alpha-dependent immune surveillance.


Overproduction of the GAL1 or GAL3 protein causes galactose-independent activation of the GAL4 protein: evidence for a new model of induction for the yeast GAL/MEL regulon. The transcriptional activation function of the Saccharomyces cerevisiae GAL4 protein is modulated by the GAL80 and GAL3 proteins. In the absence of galactose, GAL80 inhibits the function of GAL4, presumably by direct binding to the GAL4 protein. The presence of galactose triggers the relief of the GAL80 block. The key to this relief is the GAL3 protein. How GAL3 and galactose activate GAL4 is not understood, but the long-standing notion has been that a galactose derivative formed by catalytic activity of GAL3 is the inducer that interacts with GAL80 or the GAL80-GAL4 complex. Here we report that overproduction of the GAL3 protein causes constitutive expression of GAL/MEL genes in the absence of exogenous galactose. Overproduction of the GAL1 protein (galactokinase) also causes constitutivity, consistent with the observations that GAL1 is strikingly similar in amino acid sequence to GAL3 and has GAL3-like induction activity. Cells lacking the GAL10-encoded UDP-galactose-UDP-glucose epimerase retained the constitutivity response to overproduction of GAL3, making it unlikely that constitutivity is due to endogenously produced galactose. A galactose-independent mechanism of constitutivity is further indicated by the inducing properties of two newly created galactokinaseless alleles of GAL1. On the basis of these data, we propose a new model for galactose-induced activation of the GAL4 protein. This model invokes galactose-activation of the GAL3 and GAL1 proteins which in turn elicit an alteration of the GAL80-GAL4 complex to activate GAL4. This model is consistent with all the known features of the system and has important implications for manipulating GAL4-dependent transcriptional activation in vitro.


Posttranslational control of Ty1 retrotransposition occurs at the level of protein processing. High-level expression of a transpositionally competent Ty1 element fused to the inducible GAL1 promoter on a 2 microns plasmid (pGTy1) overcomes transpositional dormancy in Saccharomyces cerevisiae. To investigate the mechanisms controlling the rate of Ty1 retrotransposition, we quantitated transposition and Ty1 gene products in cells induced and uninduced for expression of pGTy1. The increase in Ty1 transposition was 45- to 125-fold greater than the increase in Ty1 RNA effected by pGTy1 induction. Translational efficiency of Ty1 RNA was not altered in transposition-induced cells, since p190TYA1-TYB1 protein synthesis increased in proportion to steady-state Ty1 RNA levels. Therefore, expression of a pGTy1 element increases the efficiency of Ty1 transposition at a posttranslational level. Galactose induction of pGTy1 enhanced TYA1 protein processing and allowed detection of processed TYB1 proteins, which are normally present at very low levels in uninduced cells. When the ability of genomic Ty1 elements to complement defined mutations in HIS3-marked pGTy1 elements was examined, mutations in the protease domain or certain mutations in the integrase domain failed to be complemented, but mutations in the reverse transcriptase domain were partially complemented by genomic Ty1 elements. Therefore, the activity of Ty1 elements in yeast cells may be limited by the availability of Ty1 protease and possibly integrase. These results suggest that Ty1 transposition is regulated at the level of protein processing and that this regulation is overcome by expression of a pGTy1 element.


Global regulation of mitochondrial biogenesis in Saccharomyces cerevisiae: ABF1 and CPF1 play opposite roles in regulating expression of the QCR8 gene, which encodes subunit VIII of the mitochondrial ubiquinol-cytochrome c oxidoreductase. The multifunctional DNA-binding proteins ABF1 and CPF1 bind in a mutually exclusive manner to the promoter region of the QCR8 gene, which encodes 11-kDa subunit VIII of the Saccharomyces cerevisiae mitochondrial ubiquinol-cytochrome c oxidoreductase (QCR). We investigated the roles that the two factors play in transcriptional regulation of this gene. To this end, the overlapping binding sites for ABF1 and CPF1 were mutated and placed in the chromosomal context of the QCR8 promoter. The effects on transcription of the QCR8 gene were analyzed both under steady-state conditions and during nutritional shifts. We found that ABF1 is required for repressed and derepressed transcription levels and for efficient induction of transcription upon escape from catabolite repression, independently of DNA replication. CPF1 acts as a negative regulator, modulating the overall induction response. Alleviation of repression through CPF1 requires passage through the S phase. Implications of these findings for the roles played by ABF1 and CPF1 in global regulation of mitochondrial biogenesis are discussed.


Long-term potentiation is associated with increases in quantal content and quantal amplitude. Long-term potentiation (LTP) of synaptic transmission in CA1 neurons of the hippocampus, elicited by the conjunction of presynaptic firing and postsynaptic depolarization, is an important model of plasticity, which may underlie memory storage. Although induction of LTP takes place in the postsynaptic cell, it is not clear whether it is expressed through an enhancement of transmitter release or through an increased postsynaptic response to the same amount of transmitter. Analysis of the trial-to-trial amplitude fluctuations of synaptic signals, that is quantal analysis, gives an important insight into the probabilistic mechanisms of transmission, although attempts to apply it to the mode of expression of LTP have so far yielded inconsistent results, at least in part because they have relied on models of transmitter release that have not been confirmed experimentally. Here we report clear evidence for quantal fluctuation in a subset of cells. Induction of LTP in these cells causes abrupt increases in either quantal content or quantal amplitude, or both. This shows that two different mechanisms can underlie the maintenance of LTP.


Unusual HLA-B alleles in two tribes of Brazilian Indians. The Kaingang and Guarani are culturally and linguistically distinct tribes of southern Brazil. Like all Amerindian groups they show limited HLA polymorphism, which probably reflects the small founder populations that colonized America by overland migration from Asia 11,000-40,000 years ago. We find the nucleotide sequences of HLA-B alleles from the Kaingang and Guarani to be distinct from those characterized in caucasian, oriental and other populations. By comparison, the HLA-A and C alleles are familiar. These results and those reported in the accompanying paper on the Waorani of Ecuador reveal that a marked evolution of HLA-B has occurred since humans first entered South America. New alleles have been formed through recombination between pre-existing alleles, not by point mutation, giving rise to distinctive diversification of HLA-B in different South American Indian tribes.


[Ubiquitin-dependent degradation and modification of proteins]. A large part of cellular proteins is in a dynamic state of turnover. Protein breakdown is responsible for essential cellular functions like modulation of key enzyme levels or removal of abnormal proteins. A major pathway for this selective proteolysis is mediated by the ubiquitin system, in which proteins are committed to degradation by their ligation to ubiquitin, a highly conserved 76 amino acid polypeptide. Recent evidence indicates that ubiquitination serves other functions besides marking proteins for destruction. As originally described for histones, the activities of several cellular proteins are reversibly regulated by ubiquitination and a successive de-ubiquitination step mediated by the activity of one or more isopeptidases.


[Otitis media with effusion as initial symptom of a parapharyngeal tumor]. Two patients are described who had unilateral otitis media with effusion (OME) as the first and only symptom of a parapharyngeal tumour. The first patient, a 77-year-old woman, was operated 14 months after the onset of the OME. A polymorphous adenocarcinoma was resected from the left parapharyngeal space. In the second patient, a 59-year-old man, the diagnosis was established 7 months after the first symptom. A pleomorphic adenoma was removed surgically from the right parapharyngeal space. In case of a persistent unilateral OME in an adult, a parapharyngeal tumour should be considered as one of the possible causes.


[Axonal polyneuropathy caused by acute multiple organ disease]. Complications involving the peripheral nervous system known as CIP have been reported during the course of prolonged sepsis and MOFS. These may be differentiated from Guillain-Barré's syndrome by their electrophysiological characteristics of axonal neuronal damage and the normality of spinal fluid tests. The Authors report and discuss a case which was brought to their attention.


[Prevalence of hepatitis C virus infection in a population of drug addicts and their heterosexual partners]. This study reports data concerning the prevalence of anti HCV in a population of 166 drug addicts and 25 partners (14 drug addicts and 11 with no history of substance abuse) of subjects proven anti HCV positive. The study was carried out in order to evaluate the importance of the haematic and sexual modes of transmission of HCV. The results obtained showed a prevalence of HCV infection in drug addicts with substantial statistic significance when compared with the prevalence in a population of donors. There is no evidence of statistically significant differences in prevalence between non-addicted partners and blood donors or between addicted partners and the general population of drug addicts. From these comparisons it is clear that haematic contact is an important source of contagion, whereas sexual contact appears to be of minor importance.


[A case of herpetic meningoencephalitis. Serological and imaging diagnosis]. The paper reports a case of HSVE observed in a 46-year-old patient with a negative anamnesis for previous HSV infection and with no objective signs of herpetic muco-cutaneous infection at time of hospitalisation. Due to the impossibility of carrying out bioptic and cultural tests, the diagnosis was based on the increased levels of specific anti-HSV1 antibodies in the liquor; these findings were compared to those found in serum, taking into account the increased antibody titres and in particular the ratio between the level of liquor and serum albumin and that of specific antibodies in both liquor and serum. The diagnostic importance of cerebral CT was confirmed by the early revelation of morphostructural alterations during the acute phase. RMN also allowed a more detailed imaging diagnosis to be made. From a therapeutic point of view, the paper confirms the value of acyclovir treatment.


Vanadate amplifies receptor-mediated accumulation of inositol trisphosphates and inhibits inositol tris- and tetrakis-phosphatase activities. Lithium ion, which inhibits hydrolytic degradation of inositol monophosphates, is the most common therapeutic agent used in the control of bipolar disorder. There exists evidence that elevated elemental vanadium levels may play an etiological role in at least some forms of manic-depression. Here we demonstrate that vanadate treatment of intact cells from several different clonal lines synergistically induces substantial augmentation in neurotransmitter receptor-mediated or growth factor receptor-triggered inositol trisphosphate accumulation in situ. Furthermore, studies done using cellular extracts indicate that effects of vanadate treatment in situ may be due to its ability to inhibit hydrolysis of inositol 1,4,5-trisphosphate inositol 1,3,4-trisphosphate, and inositol 1,3,4,5-tetrakisphosphate in vitro. These results suggest that vanadate treatment may facilitate characterization of inositol phosphate metabolism and intracellular signaling.


[Leu31, Pro34] NPY, a selective functional postjunctional agonist at neuropeptide-Y receptors in anaesthetised rats. Neuropeptide Y (NPY), a sympathetic cotransmitter, has both prejunctional and postjunctional actions in the cardiovascular system. In the bioassay system used here NPY attenuates cardiac vagal action (an indicator of prejunctional or Y2 action) and increases blood pressure (an indicator of postjunctional or Y1 action). [Leu31,Pro34]NPY has little or no prejunctional activity i.e. does not attenuate cardiac vagal action, but increases blood pressure as effectively as NPY. [Leu31,Pro34]NPY can therefore be used in functional experiments to distinguish between the two types of NPY receptor.


Alterations in the water intake caused by central inhibition of angiotensin-converting enzyme in the rat. In the present study we investigated the effects of central (i.c.v.) and subcutaneous (s.c.) injections of a 2 micrograms dose of lisinopryl, an inhibitor of angiotensin I(ANGI)-converting enzyme (CE), on water intake. I.c.v. but not s.c. injection of lisinopryl abolished drinking in response to s.c. isoprenaline (100 micrograms/kg) and significantly reduced drinking in response to 24 h water deprivation or s.c. polyethylene glycol (30% w/v, 10 ml/kg). Lisinopryl had no effect on water intake induced by cellular dehydration (s.c. injection of hypertonic saline (2 M NaCl)). These results are consistent with the hypothesis that lisinopryl acts as a CE blocking agent in the brain. The thrist challenge induced by hypotension using isoprenaline acts primarily by generating ANGII systemically and centrally. The other thirst challenges such as cellular dehydration are independent of the ANGII in the brain. This conclusion was made possible by utilizing a new CE blocking agent at a smaller dose than normally used for other ANG I-CE inhibitors.


Opioid peptides reduce synaptic transmission in the nucleus accumbens. Behavioral studies implicate the nucleus accumbens (NAcc) as a brain area pivotal for the rewarding effects of opiates like heroine and morphine. Therefore, we studied the effect of a variety of opioids on membrane properties and responses to synaptic stimulation in a slice preparation of the NAcc using intracellular recording. Superfusion of opioid peptides did not affect the membrane potential or input resistance of NAcc neurons, but significantly reduced both depolarizing and hyperpolarizing synaptic potentials. Naloxone superfusion significantly reversed the depressant effects of the mu and delta receptor agonists (but not those of the kappa agonist) on synaptic transmission, suggesting involvement of opiate receptors. These results imply that the predominant effect of opiates in NAcc is a reduction of synaptic transmission.


Kappa-opioid receptor stimulation abolishes mu- but not delta-mediated inhibitory control of spinal Met-enkephalin release. The possible opioid control through delta, mu and kappa receptors of the spinal release of Met-enkephalin-like material (MELM) was investigated in halothane-anaesthetized rats. The intrathecal perfusion of the delta agonist DTLET (10 microM) or the mu agonist DAGO (10 microM) resulted in a marked inhibition of MELM release, which could be prevented by the selective antagonists naltrindole and naloxone, respectively. Although the kappa agonist U 50488 H (10 microM) was inactive per se, it completely suppressed the inhibitory effect of DAGO, without affecting that of DTLET. As the selective kappa antagonist norbinaltorphimine blocked the action of U 50488 H, it can be concluded that kappa receptors modulate the mu- (but not the delta-) mediated feed back control of spinal enkephalinergic neurones.


Thyroid hormone affects the hydrolysis of inositol phospholipids in the rat hypothalamus. We have attempted to elucidate the effect of thyroid hormone on phospholipase C-linked inositol phospholipid hydrolysis in the rat hypothalamus. Hypothalamic slices of each animal, euthyroid control, hypothyroid, and thyroxine (T4)-supplemented hypothyroid rats were labeled with [3H]myoinositol in the presence of 5 mM LiCl, and then incubated for 60 min in KHG buffer containing either vehicle or 1 mM ouabain, a Na-K ATPase inhibitor. Hypothyroidism caused a significant increase in both basal and ouabain-stimulated accumulation of [3H]inositol phosphate ([3H]IP) in hypothalamic slices, whereas supplement with T4 to hypothyroid rats resulted in a complete restoration of hypothalamic [3H]IP formation to the value of euthyroid control. The present results indicate that thyroid hormone affects phospholipase C-linked inositol phospholipid hydrolysis in the hypothalamus, suggesting that negative feedback action of thyroid hormone may occur at a post-receptor site in the hypothalamus.


A possible role of free radicals in the transplantation of retinal pigment epithelial cells. Oxygen-derived free radicals have been implicated in tissue injury following ischemia and reperfusion (reoxygenation). It has been hypothesized that the radicals are produced during the early reperfusion stage. Recently, submacular implantation of retinal pigment epithelium cells has been reported. It is probable that during the procedure, the transplant and the 180-degree folded outer retina underwent a period of ischemia, followed by reoxygenation. We, therefore, infer that free radicals were produced during the reoxygenation stage of the procedure, injuring both tissues. We suggest that these hypotheses be investigated with the aim of improving the surgical outcome in eyes with age-related macular degeneration.


Hydroxylapatite ossicular replacement prostheses: a four-year experience. Between 1987 and 1991, I have used 215 hydroxylapatite middle ear implants, in various styles, for hearing reconstruction. The first such implants were composed entirely of hydroxylapatite. Because of intraoperative difficulties in shaping and trimming these prostheses, hybrid prostheses using Plasti-Pore were developed. For each of four implant designs (incus, incus-stapes, PORP, and TORP), the head is constructed from hydroxylapatite and the shaft from Plasti-Pore. Extrusion rate for the hybrid prostheses is low (4.3%). Hearing results from 47 patients with the hybrid hydroxylapatite prostheses, 140 patients with total hydroxylapatite prostheses, and 75 control group patients with homograft bone or Plasti-Pore prostheses were compared. A "successful" hearing result was achieved in 51.1%, 51.4%, and 60.0% of the three groups, respectively. Surgical technique for use of the new hybrid hydroxylapatite prostheses is described.


Treatment of large juvenile nasopharyngeal angiofibroma. The management of large juvenile nasopharyngeal angiofibromas with intracranial extension is controversial. We review our experience since 1980 with eighteen patients with juvenile nasopharyngeal angiofibroma. A diagnostic and treatment approach consisting of preoperative magnetic resonance imaging, embolization of feeding branches from the external carotid artery, and attempted complete resection was used in seven patients with intracranial disease since 1987. Serial magnetic resonance images were used for followup. Intracranial disease that was persistent or recurrent and demonstrated subsequent growth was irradiated (35 to 45 cGy). Extracranial tumor recurrences were reexcised. We advocate this approach as a safe and effective alternative to primary irradiation and its sequelae.


Cytomegalovirus infection of the larynx in the acquired immunodeficiency syndrome. CMV is the most important viral opportunist in patients with AIDS. Nearly all patients with AIDS have been exposed to CMV and the virus can be isolated from many. When CMV-related disease occurs in AIDS, it has usually involved the eye or gastrointestinal tract. In lesions in which CMV is the suspected etiology, evidence of direct pathogenicity of the virus should be obtained, usually histologically. Accurate diagnosis is essential because long-term therapy with a relatively toxic drug (ganciclovir) is indicated. We have presented a case of CMV infection of the larynx that responded, albeit slowly, to ganciclovir. Additional cases may arise as the prevalence of AIDS increases. Because of the potential for airway compromise and the tendency toward chronicity, CMV infection of the larynx should be of a significant concern both for the patient and the otolaryngologist.


Leishmaniasis epidemiology: all down to the DNA. Application of quantitative methods to the study of leishmaniasis epidemiology has allowed Dye (1992) to pinpoint important biological parameters which, if they could be accurately measured in the field, would contribute most to our knowledge of the spread of disease and key targets for control. Three areas in which laboratory-based research could impact most on leishmaniasis epidemiology were highlighted by Dye (1992): (i) the development of accurate diagnostic tools which can distinguish between current and past infection; (ii) to determine the underlying molecular/genetic basis to virulence polymorphisms in the parasite and study these in the context of field epidemiological studies; and (iii) to provide the molecular tools to measure genetic variation in resistance to infection in humans and in reservoir hosts of disease. This paper describes current progress in attaining these goals, highlighting first the work on isolation and field application of genomic and kDNA probes for species-specific diagnosis, and the development of PCR-based assays which can be performed under field conditions. At a more preliminary stage, studies are described in which variability in the major molecular determinants of virulence (lipophosphoglycan, GP63, and members of the HSP70 family of stress proteins) identified through studies of laboratory models of infection, is being measured in primary field isolates of Leishmania peruviana. To complete the picture, current progress in identifying and cloning the genes which control host resistance to leishmanial infection is described, along with field studies of multicase families of human disease in which linkage analysis using marker genes from the chromosomal regions bearing these genes can be used to find evidence for their role in determining disease phenotypes in man. The projected view from these studies is that the future of leishmaniasis epidemiology will be all down to the DNA.


A simple and selective method for visualizing the kinetoplast of trypanosomes by means of phosphomolybdic acid and a photographic developer. Trypanosoma cruzi epimastigotes treated with phosphomolybdic acid and the photographic developer Rodinal display bright kinetoplasts when unmounted smears are observed using low-magnification phase-contrast objectives. This simple method enables the rapid and accurate identification of kinetoplasts in trypanosomes.


Inhibition of lipid peroxidation by prostaglandin oligomeric derivatives. The inhibition of lipid peroxidation by oligomeric derivatives synthesized from prostaglandin E1 (PGE1) and PGB2 was studied using two rat models. In an in vitro model, the brain was exposed to decapitation-ischemia, the cortex was removed and homogenized, and the formation of thiobarbituric acid reactive substances (TBAR) was measured after exposing the homogenate to in vitro reoxygenation either in the presence or absence of oligomers. It was found that these oligomers could inhibit lipid peroxidation, and that their activities were higher than that of superoxide dismutase (SOD). In an in vivo administration model, either the oligomer or the vehicle was injected i.p. 30 min before decapitation. The brain was exposed to decapitation-ischemia, the cortex was homogenized and exposed to 'in vitro' reoxygenation, after which TBAR value was determined. Ester-type compounds had a greater activity than free-acid type compounds in inhibiting lipid peroxidation. A possible mechanism of the protective effect of these oligomers in ischemia/reperfusion injury may be to scavenge oxygen free radicals.


Water calorimeter dosimetry for 160 MeV protons. The absorbed dose to water from a 160 MeV proton beam as determined by a flexible, temperature regulated, sealed glass core, water calorimeter was compared to that determined from ionization chambers used in accordance with AAPM Report 16. The ratios of these doses as obtained from two experiments done over four months apart, are 0.992 +/- 0.004 and 0.990 +/- 0.004. As there are no radiation dependent parameters required for the water calorimeter, these data add to the growing body of evidence which supports the use of the calorimeter as a reliable absorbed dose standard. They also support the use of 60Co-calibrated ionization chambers used in accordance with AAPM Report 16 for the dosimetry of proton beams.


Radon and thoron associated dose to the basal layer of the skin. The radon related alpha-particle annual dose equivalent to the basal layer of the epidermis has been calculated theoretically and is estimated as 2.5 (range 1.7 to 17) mSv y-1, for the exposed, uncovered skin of the face and neck, at the UK average domestic radon exposure of 20 Bq m-3. The thoron-related annual dose equivalent is estimated as 0.3 (range 0 to 1.9) mSv y-1 at the equilibrium equivalent concentration of 0.3 Bq m-3 estimated for UK exposures. Considerably lower dose equivalents are received by regions of the skin which are habitually covered in clothing. The wide range in the dose estimates reflects the wide range in quoted plateout rates for radon and thoron daughters, and uncertainties in the magnitude of the electrostatic charge of the individual. To improve the dose estimates experimental measurements are needed of radon and thoron daughter deposition on the skin surface in situations corresponding to domestic exposure.


Fluoxetine: a review of receptor and functional effects and their clinical implications. Downregulation of serotonin 5-HT1 receptors is the most frequently reported central nervous system neural effect of subchronic exposure to fluoxetine in rodents. However, downregulation of these receptors has not been universally demonstrated. Effects of subchronic exposure on 5-HT2 receptors are mixed. Fluoxetine exposure appears to have no effect on cholinergic muscarinic receptors. Effects on beta-adrenergic receptors are controversial, as only one laboratory has reported downregulation. The majority of studies have failed to show an effect on beta-adrenergic-receptor-stimulated cAMP generation. Electrophysiologic studies support the concept that fluoxetine facilitates net serotonergic transmission through downregulation of presynaptic inhibitory autoreceptors. Data suggest that its subchronic specificity and selectivity distinguish fluoxetine from members of other classes of available antidepressants, making it a distinct therapeutic option.


Discriminative stimulus properties of the benzodiazepine receptor antagonist flumazenil. Rats were trained to discriminate the stimulus properties of the benzodiazepine (BZ) receptor antagonist flumazenil using a conditioned taste aversion procedure. On drug trials, fluid-restricted rats were injected with flumazenil (32 mg/kg), given access to a 0.25% saccharin solution for 30 min, and injected with LiCl (1.8 mEq/kg IP). On saline trials, injections of saline bracketed the period of saccharin consumption. Acquisition of the discriminated taste aversion, as measured by differential effects on drinking between saline and drug trials, developed after only five pairings of flumazenil with the LiCl injections. Flumazenil did not alter saccharin consumption in unconditioned controls (N = 9) that never received LiCl. The discrimination was also measured by flumazenil's ability to reduce the preference for saccharin over tap water using two-bottle choice tests. Flumazenil demonstrated dose-dependent generalization upon decreasing the training dose as low as 1 mg/kg. Two other BZ receptor antagonists of different chemical structure, CGS 8216 and ZK 93426, substituted completely for the flumazenil stimulus. Partial generalization was exhibited to the partial inverse agonists FG 7142 and beta-CCE, while the full inverse agonists DMCM and PTZ failed to substitute for the flumazenil stimulus. The BZ receptor agonists diazepam and alprazolam failed to substitute for the flumazenil stimulus, although partial generalization was shown with CDP. The results suggest that the BZ receptor antagonist flumazenil may produce intrinsic discriminative stimulus effects that are independent from those of BZ receptor agonists or inverse agonists.


Evidence for separate neuronal mechanisms for the discriminative stimulus and catalepsy induced by delta 9-THC in the rat. The cataleptogenic effect of delta 9-THC was compared to its discriminative stimulus effects in rats. The ED50s for the discriminative stimulus and catalepsy were 0.8 and 4.0 mg/kg, respectively, while their time courses were very similar. The ED50 of delta 9-THC for catalepsy in experimentally naive rats was not different from that in rats trained with the drug discrimination procedure, indicating that the cataleptogenic effect was not appreciably attenuated by long-term exposure to low doses of delta 9-THC. Pharmacologically, the catalepsy produced by delta 9-THC more closely resembled that of haloperidol than of morphine, since anticholinergic pretreatment eliminated the delta 9-THC-induced catalepsy while pre-treatment with naloxone had no effect. Although the cataleptogenic effect of delta 9-THC could be pharmacologically manipulated by anticholinergic pre-treatment, its discriminative stimulus effects were not changed in the same animals. These results demonstrate that distinctive mechanisms of action exist for these cannabinoid-induced behaviors.


Inhibitory effect of opiates on male rat sexual behavior may be mediated by opiate receptors outside the central nervous system. The importance of opiate receptors outside the central nervous system for the inhibitory actions of morphine on male rat sexual behavior was evaluated. Morphine (10 mg/kg) produced an almost complete inhibition of sexual behavior. This inhibition was antagonized by naloxone at a dose of 1 mg/kg but not at a dose of 0.25 mg/kg. The quaternary opioid antagonist methylnaloxone effectively blocked the inhibitory actions of morphine at a dose of 20 mg/kg but not at a dose of 5 mg/kg. Since the affinity of methylnaloxone for opiate receptors is about 5% of that of naloxone, it may be concluded that both antagonists were about equally effective in inhibiting the effects of morphine. Furthermore, the opiate-like drug loperamide was found to inhibit sexual behavior. This drug acts mainly outside the central nervous system. Its effect was blocked by both naloxone and methylnaloxone, suggesting that opiate receptors are involved. It was also shown that methylnaloxone is unable to block the reinforcing effects of morphine in the conditioned place preference procedure. Because the reinforcing effects of opiates seem to be localized to the central nervous system, it may be proposed that methylnaloxone does not antagonize morphine's central effects. Moreover, loperamide had no effect in the place preference procedure, suggesting that this drug does not act at central opioid receptors. Taken together, these data show that peripheral opioid receptors are responsible for at least some of the inhibitory actions of morphine on male sexual behavior. After treatment with morphine + methylnaloxone, ejaculatory mechanisms were facilitated, reflected in a reduced number of preejaculatory intromissions and a shortened ejaculation latency.(ABSTRACT TRUNCATED AT 250 WORDS)


Muscarinic receptors, phosphoinositide metabolism and intracellular calcium in neuronal cells. 1. We have utilised SH-SY5Y human neuroblastoma cells and primary cultures of rat neonatal cerebellar granule cells, both expressing M3 muscarinic receptors, to examine agonist driven polyphosphoinositide hydrolysis and alterations in intracellular calcium. 2. Stimulation of SH-SY5Y cells leads to a biphasic increase in intracellular calcium, the initial peak being due to the release of calcium from an intracellular store and the second maintained phase being due to calcium entry across the plasma membrane. The channel involved does not appear to be voltage sensitive, to involve a pertussis toxin sensitive G protein, or be opened by inositol polyphosphates. 3. Muscarinic receptor stimulation also leads to increased inositol polyphosphate formation in SH-SY5Y cells. Ins(1,4,5)P3 mass formation was biphasic in profile whereas Ins(1,3,4,5)P4 mass formation was slower and monophasic in profile. These data are consistent with substantial activity of 5-phosphatase (dephosphorylating Ins(1,4,5)P3 to Ins(1,4)P2) and 3-kinase (phosphorylating Ins(1,4,5)P3 to Ins(1,3,4,5)P4) in SH-SY5Y cells. 4. In order to better understand the role of Ins(1,4,5)P3 and its metabolites in calcium homeostasis we have examined the ability of a variety of natural and synthetic analogues to release intracellular sequestered calcium. The Ins(1,4,5)P3 calcium mobilizing receptor displays a remarkable degree of stereo- and positional selectivity with the most potent agonist to date being Ins(1,4,5)P3 (EC50 = 0.09 microM). 5. As an alternative to the continuous SH-SY5Y neuroblastoma (tumour derived) cell line we have used the primary cultured cerebellar granule cell. These cells also display a biphasic increase in Ins(1,4,5)P3 mass and a subsequent release of intracellular stored calcium. In our hands carbachol appears to increase calcium influx, a response which is only visible in the absence of magnesium.


Age-related differences of cholecystokinin receptor binding in the rat brain. 1. Cholecystokinin and benzodiazepine receptor binding was evaluated in 2-, 9- and 18-month old rats in the brain regions where cholecystokinin octapeptide and gamma-aminobutyric acid are known to coexist in common nerve cells (frontal cortex, hippocampus). 2. There was a regionally selective alteration of hippocampal 3H-CCK-8 binding in the oldest age group, if compared to both young and adult animals. Non-linear regression analysis of binding data revealed significantly lower apparent number of binding sites (Bmax), and twofold (but not statistically significantly) higher binding affinity for the radiolabelled ligand. No differences between any age groups in 3H-flunitrazepam binding to benzodiazepine receptors were found. 3. The results suggest that changes in cholecystokinin receptor characteristics might contribute to the behavioural impairments in aged rats.


Effect of 80 kDa protein of porcine follicular fluid on gonadotropin stimulated progesterone production in rat granulosa cells in vitro. We reported the presence of a 80 kDa polypeptide in porcine follicular fluid that inhibited the binding of 125I-radiolabelled hFSH as well as hCG to the rat ovarian gonadotropin receptors. In the present study, the biological activity of the receptor binding inhibitor is determined using an in vitro bioassay procedure. Granulosa cells isolated from PMSG primed immature rat ovaries respond to exogenously added gonadotropins in terms of progesterone production. Addition of fractions containing the gonadotropin receptor binding inhibitory activity inhibited progesterone production stimulated by the gonadotropins in a dose-dependent fashion. The receptor binding inhibitory activity was also capable of inhibiting progesterone production stimulated by PMSG, which has both FSH- and LH-like activities in rats. In contrast, progesterone production stimulated by dbcAMP was not inhibited by the receptor binding inhibitor. This result indicates that the site of action of the inhibitor is proximal to the formation of the cAMP. The above observations point out to a possible role for this factor in modulating gonadotropin activity at the ovarian level.


Alcohol and cocaine abuse. A comparison of epidemiology and clinical characteristics. Increasing numbers of persons presenting for substance abuse treatment are multiple-substance users and their predominant drugs of choice are cocaine and alcohol. There are similarities in the treatment of alcohol and cocaine abuse, but important clinical differences need to be acknowledged. This chapter compares and contrasts the most recent epidemiology and clinical characteristics of both alcohol and cocaine abusers. First, use and abuse trends in the United States for both substances are described. The clinical characteristics of dependence on either substance are contrasted in detail, and finally, treatments are briefly described. Treatment for both disorders can be accomplished in the same setting if important demographic and pharmacological differences are addressed. In either disorder, comorbid psychopathology is frequent and appropriate treatment matching based on diagnosis is imperative.


Alcohol and cocaine. Clinical and pharmacological interactions. Both clinical experience and epidemiological studies in community and specialized (e.g., treatment) populations indicate that the prevalence of co-use of alcohol and cocaine, and the comorbidity of alcoholism and cocaine addiction, are greater than would be expected from the chance occurrence of two independent conditions. Alcohol and cocaine have pharmacokinetic and pharmacodynamic interactions that may account for some of this co-use. While their reinforcing properties have neuropharmacological and behavioral differences, a unified theory of reinforcement by alcohol and cocaine has been proposed, involving dopamine activity in the ventral tegmental area-nucleus accumbens circuit. Regardless of their pharmacology, the prevalent co-use of alcohol and cocaine has important implications for drug abuse treatment and indicates the need for future research on this topic.


Hypothalamic-pituitary function during alcohol exposure and withdrawal and cocaine exposure. This chapter examines the neuroendocrine effects of acute exposure to and withdrawal from alcohol and cocaine, with special emphasis on the hypothalamic-pituitary-adrenal (HPA) axis. We present the results from two preliminary controlled inpatient studies that document HPA dysfunction during acute exposure to alcohol and cocaine and during withdrawal from alcohol. We discuss the methodological approach of these studies in comparison to related attempts in the literature to use measures of thyroid and prolactin regulation to predict risk of relapse to alcohol and cocaine use, respectively. Our data and the results of related studies are presented in the context of a proposed index of HPA axis dysfunction that may provide a useful clinical measure of susceptibility to relapse during protracted abstinence from alcohol or cocaine.


The role of alcohol in cocaine dependence. The relationships between the patterns of alcohol and cocaine use are examined using information derived from the authors' research data and from the literature. Excessive alcohol drinking was very prevalent among males seeking treatment for cocaine dependence at a Veterans Administration hospital in West Los Angeles. Fifty percent met DSM-III-R criteria for alcohol dependence. The drug-related behaviors of patients with the diagnosis of dependence on cocaine only were compared with those of patients dependent on cocaine and alcohol. The progression of the cocaine addiction was not significantly different in those two groups. However, patients with dependence on cocaine and alcohol were more likely to be users of other drugs of abuse. Concerning psychosocial behaviors, both groups maintained employment and stable interpersonal relationships throughout significant periods of their addictive career. One year after drug treatment, both subsamples showed significant improvement in terms of cocaine and alcohol use and social adjustment. Research implications and the possibility that both dependent conditions may share neurobehavioral reinforcing mechanisms are discussed.


[Effects of Tilade in asthma].

[Effects of Tilade in asthma]. When administered in a single dose, nedocromil can inhibit the effect of various stimuli: adenosine, SO2, cold air, smog, tachykinins, metabisulfite, exercise and antigen. When administered in a chronic way, this preparation diminishes bronchial hyperresponsiveness. In a clinical study done in England, nedocromil has improved asthma symptoms, PEFR and the need for inhaled beta-2-sympathomimetics. In a study carried out in Israel, nedocromil given for 6 weeks has resulted in an improvement in FEV1 of 0.5 l. In an international study, nedocromil has been preferred to placebo by asthmatic subjects and physicians. Nedocromil can therefore be proposed in: 1) mild asthma, as a long term treatment, with or without allergic causes; 2) in moderate asthma to improve symptoms; 3) in severe asthma to reduce the need for oral steroids (?) There is a need for widening the delivery system of nedocromil, which is only available in a metered-dose inhaler and to find more powerful formulations.


[Respiratory infection and bronchial cancer]. Respiratory infections are very frequent in the course of lung cancer. The diagnosis is often difficult because of the existence of a chronic inflammatory process and of radiologic abnormalities related to lung cancer. Clinical symptomatology is atypical: no clear distinction between fever due to inflammation or infection, productive cough linked with chronic bronchitis often associated.... Radiological abnormalities are difficult to interpret: infectious lobar infiltrate or atelectasis, nature of a pleural effusion.... Biological data bear little interest because of their lack of specificity. Depending on their evolution, respiratory infections have particular characteristics. Before treatment, the problem is more often related to a fever and/or a radiological abnormality. During the post-operative period, fever generally corresponds to infectious phenomena, not necessarily originating from the lung. Among patients treated by radiotherapy and/or chemotherapy, it is sometimes difficult to conclude among several diagnosis possibilities: infectious lung condition, secondary infectious sites, iatrogenic interventions. The various possibilities that can be encountered can be divided into three main categories: in the case of lung infection in a non-neutropenic patient the organisms are those of a common community-acquired respiratory infection and it is logical to suggest a treatment based on penicillin A possibly associated with an inhibitor of beta-lactamases. In the case of a localised lung infection in a neutropenic subject, it is essential to cover gram negative bacilli by adding a beta-lactam and an aminoside. Finally, in the case of interstitial lung involvement, opportunistic bacteria are likely and it is essential to propose an endoscopic examination before initiating a therapeutic scheme.(ABSTRACT TRUNCATED AT 250 WORDS)


[Basic research on nedocromil sodium].

[Basic research on nedocromil sodium]. Nedocromil sodium is a new chemical entity. This compound is very hydrophilic and is well absorbed by tissues such as the lung but not by tissues with tight junctions such as the gut. This product is chemically different from all drugs currently used for the treatment of airway diseases. The in vitro effects of nedocromil sodium are reviewed. Nedocromil sodium is capable of blocking: 1) the chemotaxis of neutrophils; 2) the activation of macrophages and monocytes by IgE; 3) the release of histamine from mast cells; 4) the cytotoxicity of platelets; 5) the release of LTC4 from eosinophils. Nedocromil sodium thus seems to have an effect on each of the cells which are implicated in the allergic reactions. In animals, nedocromil sodium can block the immediate bronchoconstriction induced by an antigen, adenosine and neurokinin A. Nedocromil sodium can also block the increase in bronchial responsiveness induced by antigen exposure. Moreover, vascular permeability induced by ovalbumin is reduced by nedocromil sodium. In summary, nedocromil sodium demonstrated a significant inhibitory effect of inflammation in both in vivo and in vitro models.


[Review of North American studies on nedocromil sodium]. Nedocromil sodium is a new prophylactic antiasthmatic medication. Its therapeutic profile is close to sodium cromoglycate but its antiinflammatory properties seem to be superior. North-American studies on this medication have shown that this preparation can inhibit the bronchial response to exercise, inhalation of cold air and to antigens. It can replace theophyllines and is a useful preparation in asthmatic subjects requiring several medications to control asthma in improving the control of asthma and reducing the need for bronchodilators. Nedocromil sodium allows for a reduction in the doses of oral steroids in some severe asthmatic subjects. Side effects are infrequent, minor and mainly consist in a feeling of bad taste.


A study of biological behavior based on the expression of a proliferating antigen in neuroendocrine tumors of the digestive system. The expression of a proliferating antigen by Ki-67 immunohistochemistry was evaluated in 32 gastrointestinal carcinoids and in 5 pancreatic islet cell tumors. In the tissue sections the number of labelled nuclei was calculated per tumor area. The tumors were classified as low proliferating (less than 0.3 labelled cells/mm2), medium proliferating (0.3-1 labelled cells/mm2), and high proliferating (greater than 1 labelled cell/mm2). In 26 tumors obtained from patients receiving antitumor therapy (alpha-interferon) the proliferative activity was decreased. In treated midgut carcinoids the proliferative activity in metastatic tissue was significantly reduced (p less than 0.05). Though not statistically significant, primary midgut carcinoids collected from untreated patients displayed a lower proliferative activity than liver metastases. A survival analysis revealed that patients with tumors displaying low proliferative activity had a better survival than those with high proliferative activity (p less than 0.05). Single cell cytofluorometric DNA analyses showed regular diploid stem cell lines in the majority of tumors from untreated patients (9/11 cases). No correlation was found between the calculated proliferative activity and the DNA profile. The obtained results indicate that the expression of a proliferation antigen by Ki-67 immunohistochemistry can be used to evaluate the biological behavior of neuroendocrine tumors of the digestive system and predict survival.


Pre- and post-synaptic effects of spiradoline and U-50488H, selective kappa opioid receptor agonists, in isolated ileum. In guinea pig ileum, spiradoline (2 x 10(-6) M or greater) and U-50488H (3 x 10(-6) M or greater) suppressed contractile responses to acetylcholine (ACh), histamine, and BaCl2. Inhibition by spiradoline (2 x 10(-5) M) of ACh-induced contractions was not antagonized by pretreatment with naloxone (3 x 10(-4) M). Spiradoline (2 x 10(-8) M or greater) and U-50488H (3 x 10(-8) M or greater) caused dose-dependent inhibition of the contractile response of guinea pig ileum to transmural electric stimulation. The inhibitory effect of spiradoline or of U-50488H at low concentrations was reduced by a high concentration of naloxone (3 x 10(-4) M). Spiradoline at low concentrations ranging from 2 x 10(-9) to 2 x 10(-7) M reduced spontaneous contractions in rabbit ileum. Naloxone (3 x 10(-4) g/ml) antagonized the spiradoline-induced inhibition, but marked inhibition by spiradoline at 10(-4) g/ml was not restored by naloxone. These results suggest that both kappa agonists exert presynaptic inhibitory action on cholinergic nerve endings in the myenteric plexus at a low concentration range of 10(-9) to 10(-7) M and directly inhibit the smooth-muscle motility of the gut at greater concentrations.


Implication of GAP in Ras-dependent transactivation of a polyoma enhancer sequence. Controversy exists as to whether the interaction of a guanosine triphosphatase activating protein (GAP) with Ras proteins functions both to initiate and to terminate Ras-dependent signaling events or only to terminate them. GAP-C, a carboxyl-terminal fragment of GAP that is sufficient to stimulate GTPase activity, inhibited the stimulation of transcription produced by some oncoproteins (v-Src, polyoma middle T, wild-type Ras, and oncogenic Ras) but not that produced by v-Mos. Wild-type GAP did not affect transcription induced by oncogenic Ras but reversed the inhibitory effect of GAP-C on transcription induced by oncogenic Ras. These results indicate that GAP is a negative regulator of wild-type Ras and elicits a downstream signal by interacting with Ras-GTP (guanosine triphosphate).


Acquisition of myogenic specificity by replacement of three amino acid residues from MyoD into E12. The basic helix-loop-helix (bHLH) protein MyoD is a transcription factor that is important for the induction of the myogenic phenotype. The DNA binding basic region (13 amino acids) is necessary for recognition of the consensus MyoD binding site, for transcriptional activation, and for conversion of fibroblasts to muscle. In contrast, the non-tissue-specific bHLH protein E12 can bind to the MyoD binding site but does not induce myogenesis. Here, it is shown that only two amino acids in the MyoD basic region and a single amino acid from the junction, which separates the basic region and helix 1, are sufficient for myogenic specificity when substituted into the corresponding region of E12. These findings suggest that the recognition of particular determinants in the basic region is required for conversion of fibroblasts to muscle.


A site on rod G protein alpha subunit that mediates effector activation. The heterotrimeric guanine nucleotide binding proteins (G proteins) are activated by sensory or hormone receptors. In turn, the G proteins activate effector proteins such as adenylyl cyclase, cyclic guanosine 3',5'-monophosphate phosphodiesterase (cGMP PDE), phospholipase C, and potassium and calcium ion channels by mechanisms that are poorly understood. A site on the alpha subunit of the G protein transducin (alpha t) has been identified that interacts with and activates cGMP phosphodiesterase, the effector enzyme in rod photoreceptors. A 22-amino acid peptide, corresponding to residues 293 to 314 from the COOH-terminal region of alpha t, fully mimicked alpha t and potently activated PDE. This region is adjacent to the receptor activation domain; thus, the alpha subunit of this G protein has a site for interaction with both its effector and receptor that maps near the COOH-terminus.


The oxidizing capacity of the earth's atmosphere: probable past and future changes. The principal oxidants in the lower atmosphere are ozone (O3) and two by-products of O3 photodissociation, the hydroxyl radical (OH) and hydrogen peroxide (H2O2). A number of critical atmospheric chemical problems depend on the earth's "oxidizing capacity," which is essentially the global burden of these oxidants. There is limited direct evidence for changes in the earth's oxidizing capacity since recent preindustrial times when, because of industrial and population growth, increasing amounts of O3 precursor trace gases (carbon monoxide, nitrogen oxides, and hydrocarbons) have been released into the atmosphere. The concentrations of O3 and possibly H2O2 have increased over large regions. Models predict that tropospheric O3 will increase approximately 0.3 to 1% per year over the next 50 years with both positive and negative trends possible for OH and H2O2. Models and the observational network for oxidants are improving, but validation of global models is still at an early stage.


Identification of the Ah receptor nuclear translocator protein (Arnt) as a component of the DNA binding form of the Ah receptor. The Ah (dioxin) receptor binds a number of widely disseminated environmental pollutants, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and polycyclic aromatic hydrocarbons, and mediates their carcinogenic effects. The ligand-bound receptor activates Cyp 1a1 gene transcription through interaction with specific DNA sequences, termed xenobiotic responsive elements (XREs). The Ah receptor nuclear translocator protein (Arnt) is required for Ah receptor function. Arnt is now shown to be a structural component of the XRE binding form of the Ah receptor. Furthermore, Arnt and the ligand-binding subunit of the receptor were extracted as a complex from the nuclei of cells treated with ligand. Arnt contains a basic helix-loop-helix motif, which may be responsible for interacting with both the XRE and the ligand-binding subunit.


Calmodulin trapping by calcium-calmodulin-dependent protein kinase. Multifunctional calcium-calmodulin-dependent protein kinase (CaM kinase) transduces transient elevations in intracellular calcium into changes in the phosphorylation state and activity of target proteins. By fluorescence emission anisotropy, the affinity of CaM kinase for dansylated calmodulin was measured and found to increase 1000 times after autophosphorylation of the threonine at position 286 of the protein. Autophosphorylation markedly slowed the release of bound calcium-calmodulin; the release time increased from less than a second to several hundred seconds. In essence, calmodulin is trapped by autophosphorylation. The shift in affinity does not occur in a site-directed mutant in which threonine at position 286 has been replaced by a non-phosphorylatable amino acid. These experiments demonstrate the existence of a new state in which calmodulin is bound to CaM kinase even though the concentration of calcium is basal. Calmodulin trapping provides for molecular potentiation of calcium transients and may enable detection of their frequency.


Renal single photon emission computed tomography: should we do it? Although single photon emission computed tomography (SPECT) imaging has established a place for itself in clinical nuclear medicine for heart and brain studies, its place in renal imaging is not yet clear. Renal SPECT has been subject to limitations imposed by the efficiency of imaging equipment, and has been confined to use with static imaging agents such as technetium-99m (99mTc) dimercaptosuccinate (DMSA). SPECT has been used to investigate space-occupying lesions and anatomical abnormalities, and for quantitative studies of renal uptake and volume. In these areas, it has provided little advantage over conventional imaging, but it has been helpful in individual cases. High-resolution SPECT is a promising new development, which may have applications in detecting and classifying renal scarring. It deserves careful evaluation.