Plasticity of neuronal connections in developing brains of mammals. Although mature nervous systems show substantial malleability following various surgical or environmental manipulations, developing brains show far more prominent plasticity, particularly in terms of morphological features. Neuronal circuits, for example, can be dramatically rewired following neonatal but not adult brain lesions. It remains unknown why neuronal circuits in developing brains show such remarkable plasticity. A number of anatomical and physiological studies suggest that there are transient projections in developing brains and they are eliminated by cell death and/or collateral elimination as development proceeds. This raises a possibility that aberrant projections observed following various surgical or environmental manipulations such as partial denervation, results from retention or stabilization of transient projections. However, evidence suggests that cell death does not play an important role in developmental fine-tuning of neuronal projections. Furthermore, although the elimination of axon collaterals takes place, individual neurons appear to elaborate axonal arbors in appropriate target areas, resulting in a net increase in the size of axonal arbor emerging from individual neurons. In accord with these observations, the number of synapses appear to increase during the period when axonal elimination proceeds. Taken together, reinforcement of appropriate projections rather than elimination of excessive connections plays a major role in developmental specification of neuronal connections. Appearance of aberrant projections after partial denervation may not be a consequence of disordered axonal growth, since they form topographic maps which precisely mirrors those for normal projections. They may be induced due to reinforcement of pre-existing neuronal connections rather than to construction of novel pathways. Observations of axonal morphology in denervated areas indicate that lesion-induced enlargement of projections is due to transformation of axonal morphology, from simple and poorly branched to multiply branched. Perhaps such simple and poorly branched axons in inappropriate target areas may represent ones in the course of elimination but they may serve as a source of sprouting when denervated. In other words, after total elimination of axons any surgical or environmental manipulation cannot induce enlargement of projections. The mechanisms underlying such modifiability of neuronal connections remains unclarified but possible participation of an activity-dependent competitive mechanism is discussed.


Spinal cord cells innervating the bilateral parabrachial nuclei in the rat. A retrograde fluorescent double-labeling study. The internal lateral nucleus (IL) of the parabrachial nucleus receives information from the spinal cord. The IL perhaps relays nociceptive signals to the intralaminar nuclei of the thalamus, apparently being implicated in the motivational-affective component of pain reactions. However, cells of origin of spinal fibers to the IL have not been investigated enough. We intended to clarify these cells, as well as their shapes, by retrograde double-labeling techniques. Fast blue and diamidino yellow dyes were injected, respectively, into the left and right ILs. The distribution of double-labeled cells was almost the same as that of single-labeled cells on both sides of the spinal cord. The total number of bilateral double-labeled cells was highest in the dorsolateral part of the lateral funiculus (DL), followed, in order, by lamina I, the dorsomedial part of the lateral funiculus (DM), lamina V and lamina VII. A few double-labeled cells were seen in laminae II-IV, VI, VIII and X. The ratio of the total number of bilateral double-labeled cells to the total number of bilateral single-labeled cells through the spinal cord was 43% in the DL, 37% in the DM, 28% in lamina V and 24% in lamina I. The ratio was 10% or less in the other remaining laminae. No marked differences were observed between the shapes of double- and single-labeled cells.


Depending on the mode of application morphine enhances or depresses somatocardiac sympathetic A- and C-reflexes in anesthetized rats. The effects of morphine on the reflex discharges in sympathetic efferents recorded from branches of the inferior cardiac nerve (ICN) were studied in rats anesthetized with alpha-chloralose and urethane. Somatocardiac sympathetic A- and C-reflexes were elicited by single shock electrical stimulation of myelinated (A) and unmyelinated (C) afferent fibers of the tibial nerve, respectively. Application of morphine either into the femoral vein or into the subarachnoid space of the cisterna magna enhanced both the A- and C-reflexes in a dose-dependent manner, while application of morphine into the intrathecal space of the lumbar spinal cord selectively inhibited C-reflexes. All effects of morphine were antagonized by naloxone. Application of morphine via the internal carotid artery to central nervous structures above the brainstem had no effect on the somatocardiac sympathetic reflexes. It is concluded that in the anesthetized rat morphine in a dose-dependent and naloxone-reversible manner (1) depresses spinal transmission of C-afferent activity, whereas (2) at the brainstem it enhances the transmission of somatocardiac sympathetic A- and C-reflexes.


Projections of vertical eye movement-related neurons in the interstitial nucleus of Cajal to the vestibular nucleus in the cat. In two alert cats, single-unit activity of neurons related to vertical eye movement was recorded in and around the interstitial nucleus of Cajal (INC), and their projections to the ipsilateral vestibular nucleus and response to stimulation of the contralateral vestibular nerve were examined. Of 62 neurons that discharged in relation to vertical eye movement, 41 increased their firing rate for downward positions and 21 for upward positions. About one third of downward-on neurons was antidromically activated by stimulation of the ipsilateral vestibular nucleus with thresholds of 36-220 microA. None of the upward-on neurons were antidromically activated. About 60% of INC neurons (22/36) responded orthodromically to stimulation of the contralateral vestibular nerve. In particular, all the downward-on neurons that projected to the ipsilateral vestibular nucleus exhibited orthodromic responses at disynaptic latencies. The results, together with our previous finding that excitatory secondary vestibular neurons carrying vertical position signals project contralaterally to the INC, suggest that downward-on INC neurons receive direct connection from these secondary vestibular neurons and send the signals back to the ipsilateral vestibular nucleus. Interstitio-vestibular interactions through these pathways may be important in the generation of vertical eye position signals.


The interstitial nucleus of Cajal is involved in generating downward fast eye movement in alert cats. We examined whether or not the interstitial nucleus of Cajal (INC) is involved in generating vertical fast eye movement. We used a gamma-aminobutyric acid (GABA) agonist (muscimol) to deactivate the INC cell bodies bilaterally in alert head-fixed cats. The INC was identified by recording vertical eye position-related burst-tonic neurons. Following muscimol infusion into the bilateral INC, the cats were unable to hold eccentric upward eye position after saccades, and the mean time constant of postsaccadic drift was 0.4 sec. In addition, downward saccades were virtually lost without clear impairment of horizontal saccades. Muscimol infusion into the Forel's field H resulted in loss of both upward and downward fast eye movement as reported previously. Bilateral electrolytic INC lesions produced results similar to those after muscimol infusion into the INC, suggesting that the cutting of passing fibers in the INC alone cannot explain the selective loss of downward fast eye movement. These results indicate that the INC itself is involved in generating downward fast eye movement. We propose that the loss of activity in possible downward burster-driving neurons in the INC can explain the selective loss of downward fast eye movement after bilateral INC lesions.


Ischemia-induced changes in PIP2 levels of gerbil hippocampus. We carried out an immunohistochemical study to detect changes in phosphatidylinositol 4,5-bisphosphate (PIP2) in gerbil hippocampus at various times after transient ischemia, using an anti-PIP2 antibody. About 24 h after transient ischemia for 5 min, an increase in the immunoreactivity was observed which was restricted to the area of CA1 pyramidal neurons. On the other hand, after less severe ischemia lasting 2 min, which did not lead to neuronal death, a decrease in PIP2 immunoreactivity was observed at about 48 h. The results indicate that levels of PIP2 following ischemia reflect dynamic changes in phosphatidylinositol (PI) turnover which may be related to neuronal degeneration.


Differential sensitivity of human fibroblasts and endothelial cells to reactive oxygen metabolites. Reactive oxygen metabolites are implicated in tissue damage, which is often followed by fibrosis. Our aim was to evaluate the sensitivity of human fibroblasts, in comparison with umbilical vein endothelial cells, to two common reactive oxygen metabolites, to superoxide produced by hypoxanthine and xanthine oxidase, and to reagent hydrogen peroxide. Depletion of the prelabeled adenine nucleotide pool, which is a sensitive index of cell damage, was used as the basis for comparison. In the presence of hypoxanthine, xanthine oxidase caused a dose-dependent nucleotide depletion, which was more pronounced in endothelial cells. After 4 h of exposure to 100 microM hypoxanthine and 80 mU/mL xanthine oxidase, fibroblasts retained 73 +/- 2% of their adenine nucleotides but endothelial cells retained only 11 +/- 2%. Hydrogen peroxide also had a larger effect on endothelial cells; after exposure to 100 microM for 30 min, adenine nucleotides retained 36 +/- 26% of their initial radioactivity in endothelial cells but 76 +/- 8% in fibroblasts. We conclude that umbilical vein endothelial cells are inherently more sensitive to the harmful effects of reactive oxygen metabolites than are fetal skin fibroblasts.


The role of dicarboxylic anion transport in the slower Ca2+ uptake in fetal cardiac sarcoplasmic reticulum. Sarcoplasmic reticulum- (SR-)mediated Ca2+ transport is slower in the fetal heart compared with the adult. Virtually all previous studies of cardiac SR Ca2+ transport were performed in the presence of oxalate, a dicarboxylic anion that is cotransported with Ca2+ in skeletal muscle SR. If anion transport is developmentally regulated in cardiac SR, this could explain, in part, the previously reported results. The purposes of this study were to establish the presence of an SR dicarboxylic anion transport process in the rabbit heart and to determine if the perinatal changes in SR Ca2+ transport occur in a dicarboxylic anion-dependent and/or independent manner. In isolated fetal and adult rabbit cardiac SR membranes, we measured Ca2+ ATPase rates and 45Ca2+ uptake in the presence of the dicarboxylic anions maleate and succinate compared with the zwitterionic buffer PIPES, to which cardiac SR is essentially impermeable. We also measured 14C-succinate uptake by fetal and adult SR membranes. Anion-independent Ca2+ ATPase activity and net 45Ca2+ uptake were significantly lower in the fetal SR membranes than in the adult. Maleate and succinate increased the Ca2+ ATPase rates in the fetal and adult SR, but the effect was significantly greater in the adult. Maleate and succinate stimulated earlier attainment of maximal net Ca2+ uptake in the fetal and adult SR, suggesting that these dicarboxylic anions stimulated the rate of Ca2+ accumulation. Maleate and succinate significantly increased the maximal net Ca2+ uptake in the adult SR, but not in the fetus. The percentage of stimulation of Ca2+ uptake by maleate and succinate was similar in the fetal and adult SR.(ABSTRACT TRUNCATED AT 250 WORDS)


Latency in vitro of varicella-zoster virus in cells derived from human fetal dorsal root ganglia. A potential in vitro model of varicella-zoster virus (VZV) latency was developed. Dissociated human dorsal root ganglion cultures were infected with VZV and maintained for 1 wk in the presence of bromovinyl arabinosyl uracil, a potent inhibitor of VZV. Seven to 21 d after removing the inhibitor (> or = 14 d after infection), the cells were trypsinized, passed to monolayers of human embryonic lung fibroblasts, and observed for VZV reactivation as indicated by typical cytopathic effects and the appearance of VZV antigens. VZV reactivated from 56% of the cultures containing both neurons and satellite cells but not from cultures specifically enriched for either neurons, satellite cells, or ganglion-derived fibroblasts. The failure to isolate VZV from cell suspensions that were sonicated before cocultivation with fibroblasts indicated that infectious VZV was not present before reactivation. Moreover, immunohistochemical and immunoprecipitation studies revealed no VZV-specific antigens in any cultures before the reactivation stimulus. VZV antigens were detected after trypsinization and cocultivation. These findings suggest that cultures containing both neurons and satellite cells provide a model system for VZV persistence that possesses many properties of a latent infection.


Intracellular cystine loading causes proximal tubule respiratory dysfunction: effect of glycine. The present study examined proximal tubular respiration in control proximal tubules and proximal tubules loaded with cystine using 2 mmol/L cystine dimethyl ester. Basal oxygen consumption was significantly less in cystine-loaded tubules (20.6 +/- 0.5 versus 12.1 +/- 0.6 nmol O2.min-1.mg protein-1, p < 0.001). In the presence of 10(-4) mol/L ouabain, an inhibitor of the NaK ATPase, oxygen consumption was 10.2 +/- 0.7 nmol O2.min-1.mg protein-1 in control tubules and 11.4 +/- 1.0 nmol O2.min-1.mg protein-1 in cystine-loaded tubules. Thus, proximal tubular intracellular cystine loading specifically inhibits oxygen metabolism directed toward transport. Compared with control proximal tubules, cystine-loaded proximal tubules also had a lower rate of O2 consumption when the cells were permeabilized to sodium with nystatin and when mitochondrial respiration was uncoupled. Glycine, an amino acid that is cytoprotective to hypoxic proximal tubule injury, ameliorated the respiratory dysfunction observed in cystine-loaded tubules.


Glycogen storage disease type IV: inherited deficiency of branching enzyme activity in cats. Glycogen storage disease type IV due to branching enzyme deficiency was found in an inbred family of Norwegian forest cats, an uncommon breed of domestic cats. Skeletal muscle, heart, and CNS degeneration were clinically apparent and histologically evident in affected cats older than 5 mo of age, but cirrhosis and hepatic failure, hallmarks of the human disorder, were absent. Beginning at or before birth, affected cats accumulated an abnormal glycogen in many tissues that was determined by histochemical, enzymatic, and spectral analysis to be a poorly branched alpha-1,4-D-glucan. Branching enzyme activity was less than 0.1 of normal in liver and muscle of affected cats and partially deficient (0.17-0.75 of normal) in muscle and leukocytes of the parents of affected cats. These data and pedigree analysis indicate that branching enzyme deficiency is a simple autosomal recessive trait in this family. This is the first reported animal model of human glycogen storage disease type IV. A breeding colony derived from a relative of the affected cats has been established.


[Autoimmune hepatitis in children].

[Autoimmune hepatitis in children]. Autoimmune hepatitis is one of the major etiologies of chronic hepatitis in children. Chronic hepatitis may present with numerous hepatic clinical manifestations, associated with extrahepatic disorders. The etiology is unknown and the disease is characterized by the auto-antibody pattern and the positive response to immunosuppressive treatment.


[Congenital myotonic dystrophy. Diagnostic difficulties]. Congenital myotonic dystrophy is the neonatal form of Steinert myotonia. It is particularly severe when the mother carries the gene. The diagnosis is made after finding the myotonia in the mother. Prenatal diagnosis can be made by researching the Steinert's disease gene by fetal DNA analysis.


[Parameningeal cervical rhabdomyosarcoma in the neonatal period]. A case of parameningeal cervical rhabdomyosarcoma with severe bone destruction is reported in a 3 month-old infant; symptoms were present at birth. The treatment consisted of exclusive intensive chemotherapy. The outcome was favourable with complete tumor regression and vertebral bone reconstruction. The child was on complete remission without sequellae two years later.


[Acute congenital monoblastic leukemia and 9;11 translocation: a case]. Acute leukemia in the newborn child is a rare event. The clinical and biological characteristics differ from those normally encountered in the older child. Tumoral syndrome and extra-medullar locations are frequently described in the literature. Many authors have noted the difficulty of diagnosis due to the immaturity of the malignant proliferation. While it is generally agreed that therapeutic abstention is justified in the leukemoid reaction in Down's syndrome, the choice is debatable in the phenotypically intact newborn. For this reason, blastic karyotype analysis is essential and may provide guidelines when considering treatment. We report on a case history of acute monoblastic leukemia with translocation 9;11 that was diagnosed at birth in a normal newborn infant. The juxtaposition of c-ets 1 protooncogene and the beta-interferon gene has been associated with this kind of cytogenetic disease and probably constitutes a model for human leukemogenesis.


[Evaluation of a prospective survey on cryptosporidiosis in a population of children hospitalized in Strasbourg]. Cryptosporidium sp is a new enteric pathogenic organism which is responsible for self-limited acute diarrhea in infants. During a prospective study we diagnosed the first pediatric cases of cryptosporidiosis in Strasbourg. This leads us to recommend that cryptosporidiosis be looked for in the etiological diagnosis of infantile diarrhea.


[Evaluation of a three-year (1988-1990) prenatal screening of malformative uropathies in the department of Puy-de-Dôme]. During a 3-year period, 93 prenatal diagnoses of kidney or urinary tract abnormalities were carried out in the French district of Puy-de-Dôme. Sixty-nine mothers were resident in this area giving an incidence of 2.8 out of 1000 births. The pregnancy was interrupted in 10 cases, there were 2 stillbirths and three infants died within two months of life. The most frequent abnormalities were: hydronephrosis (48% of cases), megaureter with or without ureter duplication (19%) and multicystic dysplasia (16%). The prenatal diagnosis was confirmed after birth in 82% of cases. Of the 56 infants with obstructive uropathies, 17 underwent a pyeloplasty within three months of life, 32 had conservative treatment, of whom 4 were operated on afterwards, and seven could not be traced. Of the seven infants who had normal ultrasound scan at birth, three had abnormal scan during the follow-up one of whom was operated on. Nephrectomy was not performed in any of the 11 cases of multicystic dysplasia: one patient was lost to follow up, three had stable lesions and in seven cases, the size of the cysts decreased.


[Iron deficiency in hospitalized infants: study of the incidence of the nutritional factor]. We report an investigation on the iron status of 209 hospitalized 3-month--2-year-old infants over a 6-month period. Hematological parameters and infant feeding practice were determined: a total of 105 infants (50.2%) were found to be iron-depleted, with (24.8%) or without (25.4%) anemia. The mode of lactation appeared to be the main determining factor in iron deficiency, as shown by the fact that during the first months, breast feeding and consumption of an iron-fortified milk formula were 50% less frequent and of shorter duration in infants with iron deficiency than in normal infants. Incorrect diet was also more frequent in iron-deficient infants. It is concluded that iron deficiency in infants could be prevented by better informing mothers in order to encourage breast-feeding and develop the use of an iron-fortified milk formula until the infant reaches the age of one year.


[Monitoring of intracranial pressure in children. Prospective fiber optic study of an intraparenchymatous system]. The authors studied the reliability and tolerance of a new intra-parenchymatous fiberoptic device for intracranial pressure monitoring in 20 neurosurgical children. The system proved to be reliable, and the measures were well correlated with clinical evolution, CT scan lesions, and the abolition of the cerebral perfusion pressure in the case of brain death. There was a minimal shift of the electric 0 after 15 days of monitoring. The tolerance was good and no hemorrhagic, infectious or epileptic complications were observed.


[Cutaneous tumors and nodules in the neonatal period]. A review of neonatal cutaneous tumors and nodules is presented, based on a 10-year experience. Pitfalls in diagnosis are especially emphasized. A biopsy should be performed in most instances when there is a clinical doubt.


Effects of a melanotropic peptide on melanoma cell growth, metastasis, and invasion. Melanocyte stimulating hormone (alpha-MSH, alpha-melanotropin),Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Ly-Pro-Va l-NH2, regulates melanogenesis within epidermal melanocytes of many animals. An MSH analogue ([Nle4,D-Phe7]alpha-MSH) that exhibits superpotency and prolonged biological activity has been synthesized, biologically characterized, and is presently in clinical trials to determine its possible clinical use in tanning of the skin. It also has potential for the diagnosis, localization, and chemotherapy of melanoma. The effects of this analogue on the growth, metastatic behavior, and invasive potential of a melanotic variant of Cloudman S-91 murine melanoma are reported here. In an intracutaneous murine model of melanoma cell tumor growth, the analogue did not increase primary tumor growth (size) after the period of administration of the peptide hormone analogue and did not affect spontaneous lung metastases. Survival times for the control and melanotropin-treated groups were similar, suggesting that overall tumor burden was not affected by treatment with the hormone analogue. Last, melanoma cell invasion through a human amniotic basement membrane in vitro was not enhanced compared to untreated cells.


The dimerization domain of LFB1/HNF1 related transcription factors: a hidden four helix bundle? LFB1/HNF1 alpha and LFB3/HNF1 beta bind DNA as dimers and form heterodimers together in vivo and in vitro. The dimerization domain has been located in both proteins in the 32 N-terminal residues. In previous papers we have described the conformational stability as determined by CD and the secondary structure by NMR studies of a peptide with the amino acid sequence of the dimerization domain of LFB1/HNF1 alpha. This study presents a more complete characterization of similar synthetic peptides spanning the LFB3/HNF1 beta dimerization domain and the alpha/beta heterodimer. The HNF1 peptides represent an example of structures which cannot be determined by NOE data alone because they are not sufficient to define one unique solution. An approach is presented which combines NMR data, the protein structure database and structural analyses according to known principles of protein structure. On this basis we are able to determine possible solutions and to identify a four helix bundle as the structure most consistent with the experimental evidence.


Discordant changes in plasma ACTH and beta-lipotropin/beta-endorphin levels in Cushing's disease patients with depression. Cushing's Disease is often associated with a depressive syndrome, with mood, vegetative, and cognitive abnormalities of variable severity. In 11 patients with (pituitary ACTH-dependent) Cushing's disease (10 women, 1 man), we studied the relationship between severity of the depressive syndrome and concordance of changes in ACTH and beta-lipotropin/beta-endorphin (beta-LPH/beta-E) levels at baseline and in response to metyrapone and dexamethasone. For each condition, blood samples were drawn at 0800h, 1200h, 1600h, and 2200h. Six patients were categorized as mildly depressed (mean [+/- SD] depressed mood score = 0.17 +/- 0.4; modified Hamilton Depression scale score = 7.6 +/- 4.5) and five as severely depressed (mean depressed mood score = 2.4 +/- 0.5; modified Hamilton Depression scale score = 15 +/- 5.6) (p < 0.05). ACTH and beta-LPH/beta-E were measured by radioimmunoassay. For each experimental condition, changes in levels were scored as concordant if the two peptides moved in parallel between sampling points. There was a relationship between greater severity of depression and more frequent discordant changes in ACTH and beta-LPH/beta-E levels: The six patients with mild depression exhibited 23 concordant and 3 discordant change patterns, while the five patients with severe depression showed 8 concordant and 15 discordant patterns. The mean percentage of concordant patterns per patient differed significantly between the two groups (mildly depressed = 90.0 +/- 16.7; severely depressed = 34.6 +/- 8.7 (p < 0.001). When each study condition was examined separately, differences in the frequency of concordance between the groups reached significance during the post-metyrapone phase and with 8.0 mg dexamethasone administration.(ABSTRACT TRUNCATED AT 250 WORDS)


Different effects of baclofen on LH and cortisol responses to naloxone in normal men. The possible involvement of GABAergic B receptors in the control of LH and ACTH/cortisol secretion in response to naloxone was evaluated in seven normal men. Subjects were tested with naloxone (4 mg IV bolus plus 10 mg infused over 2 hr) with or without previous treatment with the gamma-aminobutyric acid (GABA)-ergic B receptor agonist, baclofen (5, 10 or 15 mg PO 30 min before naloxone). In additional experiments, six normal men were tested with 15 mg baclofen or placebo 30 min before a 2-hr infusion of normal saline. Plasma cortisol levels rose 70% in response to naloxone. The naloxone-induced cortisol rise was not modified by pretreatment with baclofen (5, 10 or 15 mg). Plasma LH concentrations rose 66% in response to naloxone. When the lowest dose of baclofen (5 mg) was administered, the LH response to naloxone remained unchanged. In contrast, 10 mg baclofen produced a significant reduction, and 15 mg baclofen completely abolished the naloxone-induced LH rise. The administration of baclofen or placebo alone did not change basal plasma levels of cortisol and LH. These data suggest that, in normal men, GABA B receptors participate in the endogenous opioidergic control of LH secretion, but not of ACTH/cortisol secretion.


Influence of curing time and distance on microhardness of eight light-cured liners. Eight visible-light-activated liners were evaluated to assess the degree of polymerization by microhardness comparison. Knoop hardness number values were measured on 1.0 mm-thick specimens with varied exposure times (20, 40, 60 seconds) and distances from the curing source (0, 3, 6 mm). Statistical analysis of the nine groups within each material revealed significant differences for time and distance (P < 0.05). Application of the light for at least 40 seconds resulted in significantly higher Knoop hardness number values than specimens cured for 20 seconds. The highest Knoop hardness values were obtained if the tip of the light source was 3 mm away from the light-activated liner.


Purification of porcine heart latent protein phosphatase Fc.M. Latent protein phosphatase, Fc.M, was purified from porcine heart extracts by a procedure involving precipitation at pH 5.0, DEAE-Sephacel chromatography, ammonium sulfate fractionation, chromatography on phenyl-Sepharose, Biogel-A 0.5m and poly-L-lysine-agarose. The purified enzyme had a specific activity of 12,200 nanomoles of phosphate released from phosphorylase a/mg protein when assayed following activation by pretreatment with Mn++ and trypsin in the presence of 0.2 M NaCl. The enzyme is a heterodimer of 66 kDa composed of a catalytic (37 kDa) and a modulator (31 kDa) subunit.


Purification and characterization of porcine heart type 2A protein phosphatases. Protein phosphatases 2A1 and 2A2 were isolated from porcine heart tissue extracts by precipitation at pH 5.0 and separated by chromatography on DEAE-Sephacel. Phosphatase 2A1 was then purified to apparent homogeneity by chromatography on phenyl-Sepharose, aminohexyl-Sepharose, Sephacryl S-300, and L-tyrosine-agarose. Phosphatase 2A2 was purified to apparent homogeneity by chromatography on phenyl-Sepharose, DEAE-Sephacel, aminohexyl-Sepharose and L-tyrosine-agarose. Purified phosphatases 2A1 and 2A2 had specific activities of 2200 and 2710 nanomoles of phosphate released from phosphorylase a/mg protein, respectively. The apparent molecular weights of phosphatases 2A1 and 2A2 on gel filtration were 155 and 105 kDa, respectively. Both enzymes contain 70 and 37 kDa subunits and 2A1 also contains a 57 kDa subunit. The 37 kDa catalytic subunit (2Ac) was obtained from the purified phosphatases by treatment with room temperature ethanol followed by sucrose density gradient centrifugation or gel filtration chromatography.


[Coping with illness in bronchial cancer from the viewpoint of the patients, their relatives and caregivers. A five-level assessment]. The investigation presented in this paper applies multi-level-assessment to emotional state and coping in patients with newly diagnosed bronchogenic carcinoma. Ratings are obtained by the patients themselves, their spouses/children, their physicians, their nurses, and interviewers, independently. Differences seem to override concordances. Patients generally present themselves in a more positive way, i. e. emotionally less disturbed, better adapted, and more successfully coping, than generally perceived by their associates. Discrepancies regarding specific aspects of illness recognition and coping give information about conflicts in both doctor-patient-relationship (e. g. greater amount of trust from the patients' than from the doctors' point of view) and nurse-patient-relationship (nurses claim patients being more aggressive then patients see themselves), as well as social support. Patients and relatives agree in their ratings of certain criteria of adaptation (getting along with the disease, quality of life) and in effective treatment possibilities. On the basis of factor analyses which are computed separately for each rating level, the perceptual structures of the different raters can be elaborated on.


Lomefloxacin versus norfloxacin in the treatment of uncomplicated urinary tract infections: three-day versus seven-day treatment. The South Swedish Lolex Study Group. This randomised, double-blind, multicenter study compared the safety and efficacy of lomefloxacin and norfloxacin in adult female outpatients with uncomplicated urinary tract infections. Patients were randomly assigned to one of 3 treatment groups: 400 mg lomefloxacin once daily for 3 days (L3), 400 mg lomefloxacin once daily for 7 days (L7), or 400 mg norfloxacin twice daily for 7 days (N7). A total of 703 patients (age 17-75 years) were enrolled at 21 investigative sites in southern Sweden. Clinical and microbiological evaluations were conducted at the start, 5-9 days and 3-4 weeks post therapy. Patients with quantitative urine cultures of > or = 10(4) CFU/ml of a susceptible pathogen were considered evaluable for efficacy. Escherichia coli and Staphylococcus saprophyticus were the most commonly isolated pathogens. In both L3 and L7 groups, 196 patients and in the N7 group 195 patients met the criteria for efficacy evaluation. At the 5-9 day post-treatment evaluation, 88% of the pathogens were eradicated in the L3 group, 93% in the L7 group and 93% in the N7 group. At the 3-4 week post-treatment evaluation, 81%, 82%, and 85% of urine cultures remained negative in the L3, L7, and N7 groups, respectively. No statistically significant differences between the 3 treatment groups were noted with the exception of eradication of S. saprophyticus, for which the 7 day courses were more effective at 4-9 days post treatment. No persistent pathogen developed resistance to the study drugs. All 3 treatment regimens were equally well tolerated, except for photosensitivity reactions, which were more frequently reported in patients in the lomefloxacin groups.


Dynamic MR studies of hepatocellular carcinoma with portal vein tumor thrombosis. Gd-DTPA enhanced dynamic MR studies were evaluated in six patients with hepatocellular carcinoma associated with portal vein tumor thrombus. The portal vein tumor thrombus was clearly visualized as a low intensity lesion. The hepatic segment supplied by the portal vein showed high intensity, probably due to decreased portal blood flow and compensatory arterial blood flow. Tumors located in this segment were shown as areas of relatively low intensity compared with segmental high intensity areas. Thus dynamic MR study was found to be a useful method for differentiating the tumor from the surrounding nontumorous tissue with decreased portal blood flow.


Morbidity risk factors in human cardiac transplantation. Histoincompatibility and protracted graft ischemia entail high risk of rejection and infection. Data from the first 103 human heart transplantations performed on 100 recipients (aged 14-62 years) at a single center from November 1983 to January 1990 were analyzed in order to detect morbidity-causing risk factors. Cumulative one- and five-year graft survival was 82% and 68%. Multivariate analysis revealed three independent risk factors for early rejection, viz. HLA-DR and HLA-B mismatches and no prior cardiac surgery. Graft ischemic time exceeding 71 min was an independent risk factor for rejection, especially for moderate or severe events, and for infection. HLA-DR mismatch was an independent risk factor for moderate and severe rejection events and for infections. Finally, patients operated on because of end-stage ischemic heart disease were at significantly higher risk of rejection than those with other cardiac disorders. The study has several implications: Prospective tissue typing for cardiac transplantation and selection of donors may have an impact on graft function: Damage to the graft by prolonged ischemia may be reduced by improved organ preservation.


Useful interventional thrombolytic and anticoagulant therapy for thrombosis due to embolization (TAE) in Budd-Chiari syndrome. Transarterial embolization given for hepatoma in a patient with Budd-Chiari syndrome resulted in hepatic infarction and inferior vena cava thrombosis. Transarterial membranotomy and repeated infusion of thrombolytic agents and anticoagulants directly in the thrombus brought about improvement of the circulation surrounding the liver and IVC, and recovery from hepatic failure.


Direct formation of microcells from mitotic cells for use in chromosome transfer. Microcells, cytoplasmic fragments that contain micronuclei composed of one or a few chromosomes, can be generated directly from mitotic cells. Cytochalasin B, which causes nuclear extrusion in interphase cells, has a similar effect on the chromosomes of colcemid-blocked mitotic cells. The forces generated during centrifugation in a Percoll gradient are sufficient to separate the extruded microcells from the parent cell. The chromosomes contained in an extruded microcell form micronuclei during the process, and in all respects are comparable to microcells generated from micronucleated cells except that they are uniformly in the G1 phase of the cell replication cycle. The procedure is probably applicable to all mammalian cells that grow in culture and can be employed to make microcells for the transfer of both intact and fragmented chromosomes.


MRI of symptomatic sacral perineural cyst. Sacral perineural cyst is a relatively rare condition. To our knowledge, reports of MR findings associated with sacral perineural cyst have been limited to only six cases. We present for the first time high field MR findings in a case of sacral perineural cyst. The cyst appeared as a cystic lesion in the sacral spinal canal and had intermediate signal intensity on T1W images and high signal intensity on T2*W images compared with CSF. Slight erosion remodeling of the sacrum was also seen anteriorly. Our case was symptomatic and present with radiculopathy (sciatic pain). Surgical treatment was done to result in dramatic improvement of the sciatic pain.


[Bilateral mental neuropathy revealing prolymphocytic transformation of chronic lymphocytic leukemia]. Authors report an original case of mental neuropathy which reveal prolymphocytoid transformation of Chronic lymphocytic leukaemia. They discuss various pathogenic hypotheses which can join these two events; these are evolutive markers of bad prognosis concerning this hemopathy. Clinical, etiopathological and prognostic features of mental neuropathy associated with hemopathies are studied through this case.


Multiple elements regulate phosphoenolpyruvate carboxykinase gene expression in hepatoma hybrid cells. The phosphoenolpyruvate carboxykinase (PEPCK) gene is highly expressed in cultured rat hepatoma cells, but extinguished in hepatoma x fibroblast hybrids. Extinction of PEPCK gene expression in hybrids is a polygenic process that involves several fibroblast loci, only one of which (tissue-specific extinguisher-1, TSE1) has been characterized to date. To identify sequence elements of the PEPCK gene that are involved both in TSE1-mediated extinction and in TSE1-independent processes, we assayed expression of chimeric PEPCK transgenes in transiently and stably transfected hybrid cells. We report that TSE1 responsiveness mapped to the PEPCK CRE (cAMP response element), as shown previously for the tyrosine aminotransferase gene. This was expected from the recent identification of the TSE1 gene product as a regulatory subunit of protein kinase A. However, none of the transgenes we assayed were responsive to TSE1-independent extinction mechanisms, suggesting that these controls require DNA sequences and/or chromatin structures that were not present in the transfected reporters. The implications of these findings are discussed.


Use of low molecular weight heparin in pregnancy. In a controlled study of 15 pregnant patients undergoing therapeutic termination of pregnancy, seven received subcutaneously 5,000 anti-FXa units of low molecular weight (LMW) heparin 15 and 3 h prior to the termination, and eight patients acted as controls. Paired maternal and fetal blood samples were taken (before or immediately after the termination) for assay of heparin activity by a chromogenic anti-FXa method sensitive to levels of 0.02 anti-FXa U/ml. LMW heparin was detected in all maternal samples of the test patients but was not detected in any of the fetal samples. The use of LMW heparin as a thromboprophylactic agent was then evaluated in 11 patients who were known to have a severe thromboembolic tendency, had suffered recurrent miscarriages and had responded poorly to conventional anticoagulation (oral anticoagulant, conventional heparin). All patients receiving LMW heparin in thromboprophylactic doses completed uneventful pregnancies and gave birth to healthy babies (three for the first time) without complication. Bone density scans performed in all patients shortly after the delivery showed normal mineral mass. We conclude that LMW heparin does not cross the placental barrier, and in addition offers satisfactory antithrombotic protection for both maternal and placental circulation. In addition, this study provides preliminary data from 11 patients suggesting LMWH may not give rise to maternal osteoporosis, a finding that now needs further investigation.


Expression of urokinase and its receptor in invasive and non-invasive prostate cancer cell lines. We previously reported that extracellular matrix invasion by the prostate cancer cell lines, PC-3 and DU-145 was contingent on endogenous urokinase being bound to a specific cell surface receptor. The present study was undertaken to characterize the expression of both urokinase and its receptor in the non-invasive LNCaP and the invasive PC-3 and DU-145 prostate cells. Northern blotting indicated that the invasive PC-3 cells, which secreted 10 times more urokinase (680 ng/ml per 10(6) cells per 48 h) than DU-145 cells (63 ng/ml per 10(6) cells per 48 h), had the most abundant transcript for the plasminogen activator. This, at least, partly reflected a 3 fold amplification of the urokinase gene in the PC-3 cells. In contrast, urokinase-specific transcript could not be detected in the non-invasive LNCaP cells previously characterized as being negative for urokinase protein. Southern blotting indicated that this was not a consequence of deletion of the urokinase gene. Crosslinking of radiolabelled aminoterminal fragment of urokinase to the cell surface indicated the presence of a 51 kDa receptor in extracts of the invasive PC-3 and DU-145 cells but not in extracts of the non-invasive LNCaP cells. The amount of binding protein correlated well with binding capacities calculated by Scatchard analysis. In contrast, the steady state level of urokinase receptor transcript was a poor predictor of receptor display. PC-3 cells, which were equipped with 25,000 receptors per cell had 2.5 fold more steady state transcript than DU-145 cells which displayed 93,000 binding sites per cell.


Significance of platelet beta-adrenoceptors for platelet responses in vivo and in vitro. The significance of platelet beta-adrenoceptors for platelet responses to adrenergic stimuli in vivo and in vitro was studied in healthy volunteers. Low dose infusion of the beta-adrenoceptor agonist isoprenaline decreased platelet aggregability in vivo as measured by ex vivo filtragometry. Infusion of adrenaline, a mixed alpha- and beta-adrenoceptor agonist, increased platelet aggregability in vivo markedly, as measured by ex vivo filtragometry and plasma beta-thromboglobulin levels. Adrenaline levels were 3-4 nM in venous plasma during infusion. Both adrenaline and high dose isoprenaline elevated plasma von Willebrand factor antigen levels. beta-Blockade by propranolol did not alter our measures of platelet aggregability at rest or during adrenaline infusions, but inhibited adrenaline-induced increases in vWf:ag. In a model using filtragometry to assess platelet aggregability in whole blood in vitro, propranolol enhanced the proaggregatory actions of 5 nM, but not of 10 nM adrenaline. The present data suggest that beta-adrenoceptor stimulation can inhibit platelet function in vivo but that effects of adrenaline at high physiological concentrations are dominated by an alpha-adrenoceptor mediated proaggregatory action.


Pentamidine: a non-peptide GPIIb/IIIa antagonist--in vitro studies on platelets from humans and other species. In this paper we show that the non-peptide anti-parasite agent pentamidine is a broad spectrum anti-platelet agent with an IC50 of 1.1 microM in ADP-induced platelet aggregation in human platelet rich plasma (PRP). It had similar activity when collagen, arachidonic acid, platelet activating factor, thrombin and epinephrine were used. It had no effect on platelet intracellular cAMP levels. It inhibited 125I-fibrinogen, 125I-fibronectin and 125I-von Willebrand factor binding to ADP-activated fixed platelets with IC50 values of 160, 160 and 60 nM respectively. Pentamidine showed a high degree of species selectivity with slightly less activity in monkey and dog PRP and little activity in guinea pig, rabbit, rat and mouse PRP compared with human. This was similar to the other RGD analogues tested. This species specificity was shown to be dependent on the species of platelets and independent of the species of fibrinogen. Thus, pentamidine is a potent non-peptide inhibitor of fibrinogen binding to GPIIb/IIIa.


Antibodies to thrombomodulin induce receptor-mediated endocytosis in human saphenous vein endothelial cells. The membrane glycoprotein thrombomodulin (TM) is an essential endothelial cell (EC) cofactor, which forms a 1:1 stoichiometric complex with thrombin. Binding of thrombin to the high affinity TM receptor transforms its procoagulant activity into an anticoagulant potential, by activating protein C. The fate of TM in the presence of thrombin is still unclear: some authors claim that the thrombin-TM complex is internalized in EC, while others find this complex to be stable for at least 2 h at 37 degrees C on the EC surface. In the present study, we investigated the interactions of thrombin and Fab-fragments of anti-TM antibodies, coupled to 5 or 15 nm gold particles with saphenous vein endothelial cells. Our results demonstrate that TM can be observed both on the plasma membrane and in coated structures only in the presence of anti-TM antibodies. Addition of thrombin decreased the extent of this labeling, while in double labeling experiments, where cells were incubated with 5 nm gold coupled thrombin and 15 nm gold coupled Fab fragments of anti-TM antibodies, thrombin was cointernalized only when anti-TM antibodies were present. These results show that thrombin-TM complex is not significantly internalized in EC. The internalization of this complex induced by anti-TM antibodies could play an important role in the thrombotic complications induced by anti-EC autoantibodies.


The transplacental effect of lead compounds on inorganic pyrophosphatase in brain, liver and kidneys of newborn rats. The transplacental effect of tetraethyl lead or lead acetate on the activity of inorganic pyrophosphatase in brain, liver and kidneys of newborn rats varied with the organ, the lead compound, the dose, and the route and time of administration. Enzyme activity was usually decreased in brain and liver, suggesting adverse effects of lead on metabolism in these organs. The inorganic pyrophosphatase activity was generally increased in kidneys.


Dopamine decreases receptive field size of rod-driven horizontal cells in carp retina. Receptive field size of rod-driven horizontal cells (HCs) in the carp retina was measured by the spread of responses to the slit of light stimulus with changing the distance from the recording electrode and it was found to decay with a single exponential function. By perfusing 10 microM dopamine (DA) the length constant of rod-driven HCs was reduced to half and the response amplitude in the centre increased approximately two-fold, and the input resistance was markedly increased. This suggests that DA as a neuromodulator released from interplexiform cells could decouple the rod-driven HCs which had no direct synaptic contact with the interplexiform cells.


Gap junctions between the pedicles of macaque foveal cones. Cone terminals ("pedicles") in the fovea of macaque retina were studied in electron micrographs of serial sections. Pedicles were sheathed in glia except for small (0.2 microns 2) fenestrations, 4.8 +/- 1.7 per pedicle. At each fenestration the membranes of adjacent pedicles were contiguous and marked by an adherent junction, which in turn was invariably associated with gap junctions. There were 3.2 +/- 1.4 gap junctions per adherent junction and thus, about fifteen gap junctions per pedicle. The gap junctions were small, 1.6 x 10(-3) +/- 1.8 x 10(-3) microns 2 (mean +/- SD) and were formed indiscriminately with all neighboring pedicles. An upper bound was estimated of 170 connexons per pedicle and thus a coupling conductance of 1.7 x 10(4) pS. Available psychophysical data suggest that the junctions are uncoupled at high luminance. They may couple at lower luminance where spatial averaging would improve contrast sensitivity without cost to spatial acuity.


Treatment of visceral leishmaniasis (kala-azar) with aminosidine (= paromomycin)-antimonial combinations, a pilot study in Bihar, India. A 20 d drug regimen of aminosidine (= paromomycin) at 12 mg/kg/d in combination with sodium stibogluconate at 20 mg/kg/d proved efficacious and well-tolerated in patients with visceral leishmaniasis in the State of Bihar, India. Eighteen of 22 evaluable patients achieved an ultimate cure. The remaining 4 patients, although not cleared of parasites, had their parasite grade reduced and also improved clinically. This confirms prior findings in Kenyan patients with kala-azar, and indicates that this regimen is a valid alternative to antimonial compounds alone in the State of Bihar, where cases of kala-azar not responding to antimonial drugs and intolerant of pentamidine are increasingly recorded.


[Kala-azar acquired in Croatia].

[Kala-azar acquired in Croatia]. Six weeks after his return from a two-week vacation in Croatia a 52 year-old janitor from Graz complained of loss of appetite, fever, headache, and a 9-kg weight loss. The spleen was enlarged to 16cm as measured by sonography. Laboratory tests revealed pancytopenia, a prolonged prothrombin time and elevation of serum LDH concentration. While repeated bone marrow biopsy showed no signs of leishmaniasis, high antibody titers against leishmania antigen led to the diagnosis of kala-azar. The indirect immunofluorescent antibody test (1:128) and a haemagglutination-inhibition test (1:512) showed diagnostic elevations of titers. Therapy with pentostam led to prompt defervescence and resulted in a full recovery of the patient. After six weeks a marked decrease of antibody titers in the haemagglutination-inhibition test (1:16) could be observed. Leishmaniasis has to be considered in patients with fever of unknown origin who return from Mediterranean countries. Despite a negative bone marrow biopsy a diagnosis is possible on the basis of serological tests. This is important because effective therapy is available as illustrated by this patient and because of the fact that the disease runs a lethal course if the diagnosis is missed.


Cytophotometric assay of cytochrome oxidase, lactate dehydrogenase and glucose-6-phosphate dehydrogenase activities in human peroxidized spermatozoa. Human spermatozoa contain appreciable amounts of intracellular glutathione, which has a protective function against peroxidative degradation of spermatozoal polyunsaturated fatty acids by the NADPH-dependent glutathione peroxidase/reductase enzymatic system. The glutathione system provides a basic defense against peroxidative damage, without which the superoxide dismutase system would dominate. Since oxidative damage is said to include enzyme leakage and changes in metabolism, cytochrome oxidase and lactate dehydrogenase activities were used as indicators of the energy metabolism in unwashed and washed human spermatozoa during lipid peroxidation. Lipid peroxidation was induced by aerobic incubation of sperms in the presence of sodium ascorbate and ferrous sulphate. In addition, since NADPH concentrations influence the concentration of reduced glutathione, we studied glucose-6-phosphate dehydrogenase activity as an indicator of pentose phosphate shunt activity, the main source of NADPH. Microdensitometric measurements of the three enzymes were made by a Vickers M85a scanning microdensitometer. We found that the lipid peroxidation process greatly affects the 3 enzymatic activities examined and that seminal plasma protects against the extensive deleterious effects of lipid peroxidation.


Vimentin expression in sweat gland tumours. Immunohistochemical localization of vimentin was studied in 93 cases of sweat gland tumours using a monoclonal anti-vimentin antibody. A strong immunoreactivity of vimentin was observed in modified myoepithelial or neoplastic myoepithelial cells of mixed tumour of the skin, syringoma, and sweat gland adenoma. Tumour cells in outer layers of tubular, ductal, and duct-like structures usually showed positive staining for vimentin, which coincided with modified myoepithelial cells. All tumour cells of clear cell hydroadenoma showed positive vimentin staining. Tumour cells of the luminal border of tubulo-ductal structures of mixed tumours were rarely immunoreactive for vimentin. Positive vimentin staining of tumour cells in the outer zone of tubulo-ductal structures in sweat gland tumours may be related to reactive proliferation of modified myoepithelial cells and simultaneous growth of luminal tumour cells.


Time course of changes in long-term potentiation of synaptic transmission following subcortical deafferentation on the rat hippocampus. Brief tetanic stimulation potentiates synaptic transmission both in the CA1 and dentate area of slices cut from normal rats. This long-term potentiation (LTP) was assayed in slices made at various times from rats subjected to complete bilateral sectioning of all subcortical afferents which enter the hippocampus. Over about one week survival time, LTP is present in the CA1 region of all and also in the fascia dentata of about 50% of slices. We found no signs of LTP in the dentate area of slices cut over 8 weeks after deafferentation, while the responses were clearly potentiated in the CA1 area of the same slices. Four week was the longest period when a somewhat modified version of LTP could be produced in the subcortically deafferented dentate area. The results confirm previous reports that subcortical afferents mediate some unknown factors essential for maintenance of long-term plasticity of intrinsic synapses in the fascia dentata. This unidentified, perhaps trophic influence diminishes in about 4 weeks after severing the subcortical fibers. In contrast, maintenance of subcortical inputs are apparently not required for the LTP in the intrinsic CA1 synapses.


Postnatal laminar development of cholinergic receptors, protein kinase C and dihydropyridine-sensitive calcium antagonist binding in rat visual cortex. Effect of visual deprivation. The postnatal ontogeny of 3H-pirenzepine and 3H-oxotremorine-M binding to M1-and M2-muscarinic acetylcholine receptors, respectively, as well as 3H-nicotine binding to neuronal nicotinic acetylcholine receptors, 3H-phorbol-12,13-dibutyrate binding to protein kinase C and 3H-PN200-110 binding to dihydropyridine-sensitive calcium channels was studied in individual layers of the visual cortex in both normally raised and monocularly deprived rats (one eyelid sutured at the age of 11 days) using quantitative receptor autoradiography. Postnatal ontogeny of M1-muscarinic receptors is similar in each visual cortical layer reaching the highest receptor density at the age of 15 days, whereas M2-muscarinic binding sites increase gradually from day 7 up to day 34. Highest 3H-nicotine binding is reached in all visual cortical layers at postnatal day 15 followed by a considerable decrease in binding sites until day 25. Phorbol ester binding rises considerably from birth until the age of 15 days reaching nearly the adult value in the upper layers, whereas in layers V and VI a marked decrease in binding levels until adulthood can be observed. The developmental course of 3H-PN200-110 binding sites is similar in all visual cortical layers and exhibits a moderate rise in binding sites between postnatal days 7 and 15. Monocular deprivation results in permanent changes in the developmental profiles of phorbol ester as well as calcium antagonist binding sites, whereas the alterations in muscarinic and nicotinic cholinergic receptors following monocular deprivation are only of transient nature. The data presented suggest that acetylcholine plays a modulatory role during a certain period of early postnatal maturation of the visual cortex by affecting both cholinergic receptors and associated second messenger cascades.


Study of the usefulness and the limits of ultrasonic diagnostic criteria for diagnosis of liver tumours. We studied the usefulness and the limits of the ultrasonic diagnostic criteria for liver tumours formulated by the Japan Society of Ultrasonics in Medicine (JSUM). 226 cases with liver mass lesions were enrolled in this study. At least one of the criteria reached a value of over 80% for one of the items: sensitivity, specificity, positive predictive value, negative predictive value or overall accuracy. In the differentiation of liver tumours, sharp & smooth boundary, presence of marginal hypoechoic zone, mosaic pattern, starry anechoic area, posterior echo enhancement and lateral shadows were important for HCC. For metastatic liver cancer, potato shape, coarsely irregular boundary, presence of marginal hypoechoic zone, internal target like anecho were important features. The liver pathology of the false negative cases corresponded to: a) liver tissue completely replaced or infiltrated by HCC or metastasis. b) the non-tumour tissue and tumour tissue were isoechoic but also without marginal hypoechoic zone. c) the ultrasonograms of non tumoural areas were modified by calcification of eggs of schistosomiasis and severe fibrosis. It can be concluded that most HCC and metastatic liver cancers over 3 cm in diameter can be diagnosed correctly by the JSUM's criteria. However, complimentary image diagnosis and fine needle biopsy are important to assure the highest diagnostic score in cases with US negative and small tumours.


The Haldane effect of rabbit blood under different acid-base conditions. The Haldane effect (HE), i.e. the difference in plasma pH of oxygenated and deoxygenated blood, was investigated in rabbits over a wide range of respiratory (PCO2 2.7 to 8.0 kPa) and metabolic (BE +5 to -15 mM) acid-base conditions and compared to cats, dogs and humans. Even under standard conditions (PCO2 = 5.3 kPa, BE = O mM) and normalized to the same Hb-concentration, the HE-induced pH-difference was distinctly greater in rabbits, cats and dogs (about 0.045) than in humans (0.034 - 0.039). During respiratory and metabolic acid-base changes, the HE-induced delta pH was inversely related to PCO2 and BE. The dependency of the Haldane effect on the acid-base status can be estimated by means of a linear inverse relationship between delta pH and lgHCO3-, as originally proposed for human blood by v. Mengden et al., 1969. Correspondingly, the regression analysis of the present experimental Haldane effect data of rabbits, cats and dogs, yielded highly linear correlations as well as characteristic species-related regression coefficients. Additionally considering the influence of the Hb-concentration, a most useful tool for quantitative estimation of the Haldane effect is thus available. This is of practical importance, e.g. when determining the arterial PCO2 indirectly by the Astrup method. However, Haldane corrections based only on human blood data available so far, would lead to an underestimation of the Haldane effect in common laboratory animals. If the appropriate Haldane shift of pH for a species is not considered, the resulting PCO2 may be erroneous by up to several 100 Pa, particularly in the range of metabolic acidosis.


[Familial adenomatous polyposis: what is new for the clinician?]. Familial adenomatous polyposis (FAP) is a rare autosomal-dominant disease characterized by the development of more than 100 colorectal adenomatous polyps in young adults. In the absence of surgical intervention, colorectal cancer ineluctably develops in all affected patients. Recent progress in the isolation of the gene responsible for the disease allows to detect gene carriers before they present with symptoms attributable to polyps. Moreover, the presence of four or more lesions of congenital hypertrophy of the retinal pigment epithelium is an extracolonic manifestation of FAP allowing presymptomatic screening of this disease. An effective screening programme combined with the elaboration of a registry for FAP and prophylactic colectomy should reduce mortality related to colorectal cancer. Two other extracolonic manifestations of FAP remain major causes of death: abdominal desmoid tumors and duodenal adenocarcinoma. At this time, no effective medical or surgical therapy has been found to cure these lesions. Restorative proctocolectomy with ileal reservoir is another major advance. This procedure is now regarded as the treatment of choice for patients with FAP because radical removal of all premalignant colorectal mucosa eliminates the risk of subsequent development of a colorectal adenocarcinoma.


Brain oxygenation state: preparation of isolated perfused rat brain and near-infrared spectrophotometry. The method of blood-free isolated perfused rat brain was described, of which EEG was within the normal range. The simultaneous measurement of heme a+a3 and copper in cytochrome oxidase could be performed under the various conditions such as graded hypoxia and seizure. The relationship between these two chromophores in the brain was same to that of isolated mitochondria.


The simultaneous measurement of the redox state of cytochrome oxidase in heart and brain of rat in vivo by NIR. The mutual relationship of the redox state of cytochrome oxidase in both cardiac and brain tissues were examined by the simultaneous optical measurement. The oxygenation states of cardiac and brain tissues behaved in a same manner during progressive hypoxia. We concluded that the oxygenation state of brain completely depends on the oxygenation state of cardiac tissue which determines the cardiac output.


Cerebral oxygenation state in chemically-induced seizures in the rat--study by near infrared spectrophotometry. The hemoglobin oxygenation state and the redox state of Cyt.ox. in the rat brain during and after seizure induced by PTZ were measured by using near-infrared spectrophotometry. PTZ administration caused transient reduction of Cyt.ox. in the brain, which might be a trigger for the increase of CBF during seizure. In postictal phase, although BP remained high, Cyt.ox. was in the certain reduced state, which might be due to A-V shunt. Hypoxic loading during seizure caused more reduction of Cyt.ox. than under non-epileptic conditions, which meant that seizure even under mild hypoxic conditions could cause severe hypoxic brain damage.


Nuclear magnetic resonance spectroscopy and the study of tissue oxygen metabolism: a review. Nuclear magnetic resonance (NMR) techniques are being increasingly utilised as an in vivo method to monitor tissue oxygen concentration in various organs. In muscle and heart, proton NMR spectroscopy of myoglobin has been used to calculate local oxygen tension through the oxygen sensitivity of the histidine group signal intensity. Similarly, spin lattice relaxation times of perfluorocarbon emulsions are oxygen sensitive, and this property has been taken advantage of to produce oxygen maps of brain by fluorine NMR imaging. Phosphorus NMR spectroscopy has also been extensively used to monitor bioenergetic state, which under some conditions, is directly related to tissue oxygen tension. This review will focus on these NMR techniques for oxygen determination, and will critically assess their utility for further studies.


Influence of anti-inflammatory drugs and free radical scavengers on intestinal ischemia induced oxidative tissue damage. The influence of oxygen free radical scavengers and anti-inflammatory drugs on postischemic lipid peroxidation and myeloperoxidase activity was shown. The best results were obtained from vitamin E and the antiinflammatory treatment with CP and SUL, whereas an iron elimination only showed slight effects on myeloperoxidase activity above all. In experiments without therapy a linear increase of lipid peroxides dependent on reperfusion durance was found, whereas myeloperoxidase already showed a remarkable increase during ischemia and early reperfusion. This difference can be interpreted by scavenging mechanisms, which are overcharged after an appointed durance of reperfusion.


Influence of free radical scavengers on myeloperoxidase activity and lipid peroxidation in acute skin grafts. Activated neutrophil granulocytes are an important source of oxygen free radicals in acute skin grafts. Lipid peroxidation and immigration of PMNs, indicating inflammatory mechanisms, affect each other to a variable extent. The effect of the scavengers allopurinol and alpha-D-tocopherol on both lipid peroxidation and neutrophilic infiltration was investigated. A possible mechanism for the superior effect of allopurinol compared to vitamin E was discussed.


Acute respiratory failure during pneumonia induced by Sendai virus. In this study a model of acute respiratory failure due to viral pneumonia in rats, closely resembling ARDS, is presented. Severe respiratory failure with lethal outcome in four days was induced by infection concentrated Sendai virus aerosol. This model permits evaluation of different therapeutical approaches for improving gas exchange during ARDS. Furthermore, preliminary results of surfactant substitution therapy in this model are presented.


Monitoring of redox-state of respiratory enzymes and myoglobin oxygenation in the working rat heart in normoxia and oxygen deficiency. The cellular oxygen supply in the isolated, hemoglobin-free perfused, working rat heart can be determined by measurements of myoglobin oxygenation. However, for a precise analysis of mitochondrial hypoxia and anoxia (pO2 < 0.01 Torr) redox-state of respiratory enzymes must be known. By use of the EMPHO (Frank et al. 1989) it is possible to perform a high speed spectrometry within very small tissue volumes. Because of the characteristic absorption spectra of oxygenated and deoxygenated myoglobin and of the oxidized and reduced cytochrome aa3 within the wavelength interval from 500 to 630 nm it is possible to isolate these two pigments from the remission spectra and to determine the oxygenation state of myoglobin and the redox-state of cytochrom aa3.


[Superficial and pedunculated signet ring cell carcinoma of the urinary bladder: a case report]. A case of primary signet ring cell carcinoma of the urinary bladder is described. A 63-year-old man presenting with difficulty of urination and miction pain had a pedunculated soybean-size tumor on the left lateral wall of the bladder. Specimens of the tumor were obtained by transurethral resection and the pathological diagnosis was signet ring cell carcinoma. There was no evidence of bladder metastasis from other organs. The patient then had intraoperative radiotherapy and he is alive without recurrence 20 months after the operation. We briefly discuss 73 cases of signet ring cell carcinoma of the urinary bladder collected from the English and Japanese literature. The tumor in this patient was the smallest of all cases reported previously.


Calmodulin antagonism and inhibition of erythrocyte plasma membrane Ca(2+)-pump by nifedipine, a calcium channel blocker. Nifedipine, a 1,4-dihydropyridine Ca(2+)-channel blocker inhibits calmodulin-stimulated activity of erythrocyte plasma membrane (Ca(2+) + Mg2+)-ATPase. Its inhibition of the basal activity of the enzyme increases on pre-incubation with membranes. The maximum velocity, affinity for ATP and Ca(2+)-sensitivity of pump are reduced by micromolar amounts of the antihypertensive drug. These results suggest that 1,4-dihydropyridines may interact with calcium-binding proteins and calcium-dependent enzymes at their calcium-binding sites.


Defective mitochondrial cation transport during dietary protein deficiency in rats. Mitochondria were isolated from the livers of male weanling and adult Wistar strain albino rats fed low-protein diet ad libitum for 30 and 84 days, respectively. Control animals consisted of adult and weanling rats fed purina chow for the same periods. Spectrophotometric estimations of mitochondrial passive transport of protons by following changes in matrix volume at 520 nm revealed that the mitochondria of protein-deficient rats are more permeable to protons than those of control rats; the mitochondria of protein-deficient weanling rats being more permeable than mitochondria of protein of malnourished adult rats. Similar results were obtained when a sensitive pH-glass electrode was used to monitor mitochondrial proton translocation. Although protons were slowly ejected from the mitochondrial matrix of protein-deficient rats, there was a significantly high rate of influx of the ion into the matrix of these mitochondria when compared with controls. Furthermore, FCCP, a classical protonophore carried protons less rapidly across the mitochondrial membrane of malnourished animals thus indicating that a small pH difference probably exists across the mitochondrial energy-coupling membrane of protein-deficient rats. Consequently, the rates of mitochondrial Ca(2+)-translocation were lower in protein-deficient animals than in normal rats. These defects are probably due to diet-induced looseness of mitochondrial bi-layer structure.


Acute hemorrhagic conjunctivitis due to enterovirus 70 in American Samoa: serum-neutralizing antibodies and sex-specific protection. Acute hemorrhagic conjunctivitis due to enterovirus 70 has caused extensive outbreaks in tropical areas since 1969. Between December 1, 1990, and March 4, 1991, an outbreak of acute hemorrhagic conjunctivitis due to enterovirus 70 occurred in American Samoa, where an outbreak due to the same agent had occurred in 1981. A survey of 5% of the households (134 households, 1,095 individuals) was conducted throughout the island of Tutuila. The outbreak affected 58% of the population, with age-specific attack rates greater than 50% for all age groups except children younger than 2 years. Attack rates were significantly higher for children 2-10 years old (65%) than in the remainder of the population. Women aged 21-40 years had higher rates than did men the same age (66 vs. 49%), possibly because of the close association of women and young children. At higher preepidemic titers, there was evidence of protection from clinical disease among males but not among females. Enterovirus 70 can cause large outbreaks even in a population already exposed in a previous large outbreak; protection due to previous infection is only partial.


Treatment with gallium nitrate: evidence for interference with iron metabolism in vivo. Gallium, when bound to transferrin, has been previously shown to cause tumor cell cytotoxicity by preventing cellular uptake of transferrin bound iron in vitro. Patients treated with constant infusion gallium nitrate for carcinoma show a rise in serum iron within 6 hr of the start of treatment. Serum iron returns to baseline by 24 hr post-infusion. Atomic analysis of iron and gallium content of Sephadex G-150 fractions of treatment sera indicate that about an equimolar amount of gallium and iron are associated with transferrin. These gallium and iron concentrations result in inhibition of transferrin mediated iron uptake in vitro, and in vivo allow for > 90% saturation of transferrin with metal. All seven patients who completed two courses of gallium therapy exhibited hypochromic microcytic anemia (mean fall in hemoglobin 3.5 grams %). Evidence for red cell iron depletion was confirmed by an increase (mean 3.3-fold) in zinc protoporphyrin levels. Since transferrin receptor increases on gallium treated iron requiring cells in vitro, we assessed cell surface transferrin receptor on peripheral blood lymphocytes by measuring fluorescent transferrin receptor antibody binding. A population of highly transferrin receptor positive cells peaks at 48 hr into the infusion. DNA analysis as well as double staining indicate the majority of transferrin receptor positive cells are unstimulated B lymphocytes. These studies provide the first documentation that constant infusion gallium treatment results in significant interference with iron metabolism and evidence for tissue iron depletion in vivo. These changes may correlate with therapeutic effects of gallium such as tumor response.


Is persistent polyclonal B lymphocytosis caused by Epstein-Barr virus? A study with polymerase chain reaction and in situ hybridization. Previous reports of patients with persistent polyclonal B lymphocytosis have found associations with female sex, cigarette smoking, HLA-DR7 phenotype, and moderate elevation of peripheral blood polyclonal B lymphocytes. The presence of binucleated cells and atypical lymphocytes in the peripheral blood of these patients was highly suggestive of a viral infection, such as with the Epstein-Barr virus. We report a 47-year-old asymptomatic woman who was incidentally found to have persistent polyclonal B lymphocytosis and serum IgG against virus capsid antigen (VCA) and Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) of EBV. The presence of EBV was investigated in the peripheral blood lymphocytes from this patient by in situ hybridization and polymerase chain reaction methods. EBV DNA was demonstrated in the lymphocyte fraction by polymerase chain reaction, and it was further located in lymphoid cells by in situ hybridization. These results indicate that persistent polyclonal B lymphocytosis is strongly associated with EBV.


The beta subunit modulates potassium activation of the Na-K pump. We recently cloned the alpha 1 and the beta 1 and beta 3 subunits of the Na,K-ATPase of the toad Bufo marinus. To investigate possible functional differences between beta 1 and beta 3, we studied the potassium activation of Na-K pumps expressed in the oocyte of Xenopus laevis. Na-K pump activity was measured as K(+)-induced current in voltage-clamped oocytes. We could take advantage of the relative resistance to ouabain conferred by the Bufo alpha subunit to study specifically the exogenously expressed Na-K pumps after inhibition of the ouabain-sensitive endogenous Xenopus Na-K pumps. Coinjection of Bufo alpha 1 subunit cRNA with either beta 1 or beta 3 cRNAs results in the expression of functional Na-K pumps that share similar low ouabain sensitivity but differ in their K+ half activation constant (K1/2). Similar results were obtained with Xenopus alpha 1 and beta 1 or beta 3 subunits and with Bufo/Xenopus heterodimers. We conclude that some specific sequence of the beta subunit can influence the activation of the Na,K pump by extracellular K+ ions.


Physiologic rationale for multiple sodium pump isoforms. Differential regulation of alpha 1 vs alpha 2 by ionic stimuli. A number of important themes emerge from our compartmental analyses of Na,K-ATPase biosynthesis in response to ionic stimuli. The ubiquitous alpha 1 beta 1 type sodium pump evolved to generate and maintain transmembrane Na+ and K+ gradients, and there are cell-type specific mechanisms of increasing synthesis and decreasing degradation to control surface expression of this important "housekeeping" enzyme. Expression of alpha 2 beta-type sodium pumps may have evolved in cells designated as K+ storehouses to facilitate maintenance of extracellular K+ in the presence of K+ restriction. Finally, the specialized distribution of Na,K-ATPase (and related E1-E2 type pumps) along the renal epithelia allows for monitoring and fine control of extracellular K+ and Na+ (volume). Many interesting questions remain to be answered, and we now have the probes and techniques needed to answer them.


Topology and sites in the H,K-ATPase.

Topology and sites in the H,K-ATPase. Understanding the membrane topology of the EP-type pumps has been approached largely by analysis of hydrophobicity plots, which are confusing in the COOH-terminal third of the proteins. Each pair of predicted membrane-spanning segments with the extracytoplasmic loop contains at least one cysteine, allowing fluorescent labeling of these regions of the enzymes by cysteine reagents once the cytoplasmic domain has been removed. The membrane segment arrangement of the H,K and sr Ca ATPases was investigated by tryptic cleavage of intact cytoplasmic face-out vesicles. This was followed by fluorescein or coumarin maleimide labeling of the SDS solubilized residual membrane fragments, tricine gradient gel separation, and sequencing. The presence of four membrane-spanning pairs was demonstrated for the alpha subunit of the H,K-ATPase, with no membrane retention of H9 and H10, although H9 has four cysteines based on cDNA sequencing. A similar observation was made for the Ca pump, except that fluorescein-labeled H9 was detected in the membrane with a molecular weight of 4 kD, showing that cleavage had occurred at lys958 predicted to be extracytoplasmic in a 10 membrane segment model. It seems likely that for both these enzymes the membrane domain contains only 8 alpha helical spanning segments. Cleavage at ala236 in the beta subunit was found only in leaky, not in ion-tight vesicles, arguing for a single membrane segment in this subunit. In the H,K-ATPase additional evidence for the presence and arrangement of the first, third, and fourth pair of segments was obtained by labeling the intact enzyme with extracytoplasmic inhibitory reagents. The K competitive reagent, an imidazopyridine, MeDAZIP+, labeled the first pair of membrane segments. The acid-activated SH reagent class, the pyridinyl methyl sulfinyl benzimidazoles, labeled cysteines 813 and 822 in the M5/M6 region as well as cysteine 892 in the extracytoplasmic loop between M7 and M8. No labeling of the beta subunit was found, indicating the presence of three disulfide bonds in the extracytoplasmic domain of this subunit. Both sets of extracytoplasmic reagents are predicted to bind close to the fatty acid/phospholipid head group interface. Inhibition by these reagents shows that conformational changes are transmitted between cytoplasmic and extracytoplasmic domains.


Ca(2+)-transport ATPases and their regulation in muscle and brain. Eukaryotic cells express one or more isoforms of a sarco(endo)plasmic reticulum (SERCA) and of a plasma membrane (PMCA) Ca2+ pump. Both the SERCA and PMCA gene transcripts are subject to alternative processing in a differentiation stage-dependent and tissue-dependent manner. The Ca2+ pump isoforms thus generated may present different functional properties. This is exemplified by the SERCA2a and SERCA2b isoforms which differ in their Ca2+ sensitivity. Analysis of the cDNA structures for PMCA1 predicts protein isoforms with variant calmodulin- and phospholipid-binding domains. A comparative study of the tissue-specific mechanisms governing SERCA-PMCA transcript processing and a more detailed study of the functional implication of the PMCA pumps isoform diversity will be challenging subjects for future studies.


Modulation of cellular signals by calpain. Calpain, an inactive proenzyme, translocates from the cytosol to the membrane upon binding calcium, and is activated at the membrane in the presence of calcium and PIP2. Activated calpain is very unstable and presumably used only once. Thus the primary targets of calpain are considered to be membrane or membrane-associated proteins. Activation of protein kinase C (PKC) occurs concomitantly with calpain at the membrane. Calpain hydrolyzes only the active PKC species leading to downregulation. Calpain participates in the transcriptional regulation by controlling the levels of transcription factors, c-Jun and c-Fos. The calpain gene is a TPA-responsive gene and its expression is stimulated by activation of PKC. Modulation of cellular signal transduction by controlling the levels of the component proteins, such as PKC, c-Jun and c-Fos is one of the important physiological roles of calpain.


[Microcrystal deposit arthropathies of the wrist]. Microcrystalline arthropathies of the wrist include three main forms of microcrystalline pathology: gout, chondrocalcinosis and apatite calcifications. Gout of the wrist is now rare and may be missed, particularly in its chronic form, occasionally associated with tophi and large, often asymmetrical erosions on the X-rays. Chondrocalcinosis is frequent and easy to diagnose in the wrist and may be responsible for a very typical arthropathy predominantly involving the radiocarpal joint, while the frequency and specificity of scapho-trapezoid involvement are controversial. Apatite deposits in the wrist involve various tendons and ligaments, especially the tendon of flexor carpi ulnaris, but may also be responsible for intra-articular pathology.


The expression of functional MSH receptors on cultured human melanocytes. Although alpha-MSH increases skin darkening in humans, there are several reports that it fails to have melanogenic effects on human melanocytes in vitro. The purpose of this study was to see whether cultured human melanocytes express MSH receptors. Human melanocytes were grown in the absence of artificial mitogens such as 12-O-tetradecanoyl phorbol-13-acetate (TPA) and cholera toxin (CT) and incubated for 2 h at room temperature with increasing amounts of 125I-labelled Nle4DPhe7-alpha-MSH with and without excess cold peptide. Binding was saturable and specific: Scatchard analysis gave a Kd of 4.9 x 10(-11) M and approximately 700 binding sites/cell. Human keratinocytes and fibroblasts showed no specific binding. The addition of 1 mM dibutyryl cAMP to the culture medium caused a 62% increase in MSH binding to human melanocytes. A smaller increase (25%) was seen with 10(-9) M CT while 25 mM TPA caused a 24% decrease. These results show that human melanocytes in culture express MSH receptors and that this expression can be modulated by mitogens.


[Diagnostic and therapeutic problems posed by malignant non Hodgkin lymphoma of renal origin in children. Apropos of 7 cases]. The kidneys of non Hodgkin lymphoma patients frequently contain lymphoma cells, but these tumors rarely arise in the renal tissue and are rarely located there. A diagnosis of non Hodgkin lymphoma of renal origin or predominantly located in kidneys was made in 7 patients aged 2-14 years old. These patients formed part of a total of 450 patients with non Hodgkin lymphomas seen from 1974 to 1987. The first manifestation in these 7 children was an abdominal mass associated with hypertension in 3 cases; 2 of whom presented with acute kidney failure. Ultrasonography showed hypoechogenous masses in one (2 cases) or both kidneys (5 cases). The diagnosis of malignant lymphoma was made directly in only one patient who also presented with mediastinal and abdominal lymph node enlargement. In the others, the first diagnosis was Wilms tumor (5 cases) and polycystic disease of the kidney (1 case); the correct diagnosis of malignant lymphoma was later made from biopsies of renal (4 patients) or extrarenal involved tissue (2 patients). Six of the 7 cases were Burkitt lymphomas, and all 7 were in stage III (3 patients) and IV (4 patients). 6 patients (3 stage III and 3 stage IV) were successfully treated by chemotherapy with a median follow-up of 9 years. Differential diagnosis between non Hodgkin lymphomas of renal origin and a Wilms tumor may be difficult even though kidney failure is more frequent in lymphoma. Sonography is the best method for diagnosis showing typical hypoechogenous masses and diffuse homogeneous infiltration of the kidneys. In the absence of extrarenal (meningeal, testis, bone marrow) dissemination, definite evidence of lymphoma depends on the histological examination of tumoral tissue obtained by surgical biopsy.


Antibody responses to avian encephalomyelitis virus vaccines when administered by different routes. Antibody responses to a commercial avian encephalomyelitis virus (AEV) vaccine administered by different routes were measured by an enzyme-linked immunosorbent assay (ELISA). Responses to single doses of vaccine administered by the ocular route to 10% of a flock were comparable with those obtained when all birds received a single dose in the drinking water. However, ocular vaccination of 5% of the flock resulted in significantly lower responses than those obtained when 10% were vaccinated. Maternal antibody was shown by the ELISA to persist in chickens from vaccinated flocks for up to 21 days after hatching. Day-old chickens with serum absorbances of < 0.3 at 492 nm, as determined by the ELISA, were shown to be susceptible to intracerebral challenge with the neurotropic Van Roekel strain of AEV.


The early triiodothyronine-induced changes in state IV respiration is not regulated by the proton permeability of the mitochondrial inner membrane. The time course of changes in mitochondrial respiration, alpha-glycerophosphate dehydrogenase activity, and inner membrane proton permeability and the effects of protein synthesis inhibition was determined in hypothyroid rats treated with triiodothyronine. Respiratory rates, alpha-glycerophosphate dehydrogenase activity and proton permeability were decreased in hypothyroid rats and rose to euthyroid levels 9-12 hours after triiodothyronine treatment. Some rats received actinomycin D with the hormone to inhibit protein synthesis. Sixteen hours after actinomycin D and hormone treatment, the increases in respiratory rates and in alpha-glycerophosphate dehydrogenase activity, were completely inhibited. However, actinomycin D did not inhibit the increase in the membrane's permeability.


Effect of rebamipide on mucus secretion by endogenous prostaglandin-independent mechanism in rat gastric mucosa. The effect of rebamipide (2-(4-chlorobenzoylamino)-3-[2 (1H)-quinolinon-4-yl]-propionic acid, OPC-12759, CAS 11911-87-6), an anti-ulcer agent developed to enhance defensive factors in the gastric mucosa, on gastric mucus secretion was studied by a newly developed biochemical method to measure the gastric mucus glycoprotein content in rats. 1 h after intraperitoneal administration, rebamipide did not produce a significant change in the intramucosal mucus contents (surface mucosal layer and deep mucosal layers of corpus and antral regions) but significantly increased the content of soluble mucus, which recovered from the gastric contents, to about 160% of the control value. Since rebamipide has been shown to increase the biosynthesis of prostaglandins, indometacin was administered to the rats prior to rebamipide administration to examine whether the increase in prostaglandin biosynthesis contributes to the rebamipide-induced increase in gastric mucus secretion. The increase in the soluble mucus was not altered by the pretreatment with indometacin, thus indicating that rebamipide per se has a potential to increase the gastric mucus secretion by a mechanism that is not mediated by the endogenous prostaglandins. The effect of rebamipide on the gastric mucus secretion might contribute to heal and to prevent the recurrence of peptic ulcer diseases as well as to maintain the homeostasis of the gastric mucosa.


Pharmacological properties of the novel highly potent diuretic 7-chloro-2,3-dihydro-1-(2-methylbenzoyl)-4(1H)-quinolinone 4-oxime-O-sulfonic acid potassium salt. 7-Chloro-2,3-dihydro-1-(2-methylbenzoyl)-4(1H)-quinolinone 4-oxime-O-sulfonic acid potassium salt (M17055, CAS 114417-20-8) showed potent diuretic and saluretic effects dose-dependently, in rats (p.o.), mice (p.o.) and dogs (i.v.), at doses of 0.1-100 mg/kg, 0.3-100 mg/kg and 0.01-30 mg/kg, respectively. The efficacy of M17055 for diuresis, natriuresis and chloruresis was much higher than that of hydrochlorothiazide and almost the same as that of furosemide. These results indicate that this compound may be classified as a "high ceiling diuretic". The potencies of M17055 for natriuresis in rats (p.o.), mice (p.o.) and dogs (i.v.) calculated with ED50 values were 38, 34 and 24 times, respectively, more potent than those of furosemide. Urinary excretions of sodium, chloride and potassium increased in parallel with urinary volume with the administration of M17055 or furosemide, whereas an apparent dissociation with urinary calcium and sodium excretion was observed with M17055 alone. In rats, the increase of urinary calcium excretion with M17055 was significantly lower than that with furosemide under comparable conditions of natriuresis. Moreover, in mice, M17055 decreased urinary calcium excretion at doses with low effectiveness. In clearance studies using anesthetized dogs, M17055 suppressed negative free water clearance (CH2O) under saline loaded conditions, and it decreased positive CH2O under water diuretic conditions. These changes in the effects on CH2O induced by M17055 resemble those of loop diuretics. However, M17055 could not shift negative CH2O to positive, while furosemide was able to do so. Moreover, positive CH2O decreased to nearly zero with M17055, while urine remained dilute with furosemide even at 30 mg/kg.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of praziquantel treatment on brain and kidney ATPase activities in healthy and schistosoma mansoni-infected mice. Na(+)-K(+)-dependent ouabain-sensitive ATPase and Mg(2+)-ATPase activities have been assayed in brain and kidney of healthy and Schistosoma mansoni-infected mice before and after praziquantel (EMBAY 8440, CAS 5526874-1) treatment. Schistosoma mansoni infection caused a moderate decrease of brain Na(+)-K(+)-ATPase activity with a marked inhibition of its Mg-stimulated ATPase. Meanwhile, a marked inhibition in both renal ATPase activities was observed in infected mice. Treatment of the infected mice with praziquantel (2 x 500 mg/kg b.wt.) reversed the inhibitory effect of infection on brain and kidney ATPase, so that the activity of both renal ATPase and that of brain Mg-ATPase were nearly corrected while the activity of brain Na(+)-K(+)-ATPase was increased to an extent greater than normal values. However, treatment of normal mice with praziquantel did not affect any of the measured parameters. Possible explanations of these findings are given.


[The receptor binding of 3H-corticosterone by rat hepatocytes]. The binding of 3H-corticosterone was studied on rat hepatocytes both in presence of unlabeled corticosterone, obsidan and their absence at 0 degrees-4 degrees C. The analysis of binding by the method of Scatchard showed that there are two types of specific binding sites for 3H-corticosterone. Possible existence of proper glucocorticoid receptors (Ka = 4 x 10(9)M-1, n = 0.52 x 10(-14) mol/mg prot.) has been shown, as well as possibility of 3H-corticosterone interaction with beta-adrenoreceptors (Ka = 1.2 x 10(9)M-1, n = 0.9 x 10(-14) mol/mg prot.) have been demonstrated on hepatocytes.


[The role of myeloperoxidase, a product of the polymorphonuclear leukocytes, in the pathogenesis of cataract]. The capacity of myeloperoxidase which is the product of polymorphonuclear leucocytes to induce the lens opacity was studied in young and old rabbits. It was found that the injection of myeloperoxidase solution into anterior chamber of the eye causes the irreversible lens opacification in old rabbits, not in young ones. Light microscopy of the lens section has shown the following alterations: the local thickening of the anterior capsule, disorderly accumulation of epithelial cells, formation of so-called "bladder cells" under the lens epithelium. Changes in experimental eyes are typical for cataract.


[Ganglioside protection of the erythrocyte membranes in myocardial ischemia]. Myocardial ischemia was shown to lead to modification of structural and functional organization of rat erythrocyte membranes. Thus, it was found that the activity of Na+, K+-ATP-ase markedly decreased, while accumulation of LPO products and of lysophosphatidylcholine (lyso--PC) took place in erythrocyte membranes of rats subjected to myocardial ischemia. Using nonpenetrating modifier trinitrobenzosulfonic acid, an increase in the content of modified phosphatidylethanolamine in erythrocyte membranes of ischemic rats was revealed as compared to the membranes of control animals. The intravenous administration of gangliosides (30 mg/kg) resulted in partial normalization of Na+, K+(-)ATPase activity, of LPO product and lysoPC content and of transbilayer distribution of lipids.


Inhibition of membrane redox activity by rhein and adriamycin in human glioma cells. The effect of the combination of adriamycin (ADM) with the anti-inflammatory drug rhein (RH) on the membrane redox activity in human glioma cells was investigated. RH, although less effective than ADM, inhibits ferricyanide reduction by human glioma cells in a dose-dependent manner as well as ferricyanide-induced proton release. The inhibition of the plasma membrane redox system might represent a further mechanism by which RH, other than ATP depletion, affects cell survival. The analysis of the interaction between ADM and RH, performed with the isobolar method, demonstrates a strong synergic response, probably due to an effect on different sites of action. The synergism of the ADM-RH association allows us to achieve a pre-established extent of inhibition with ADM concentrations much lower than with ADM alone. RH might, therefore, represent a very useful tool to improve the therapeutic index of ADM and to lower its general toxicity.


Effect of a hypoxic tumor cell cytotoxic disulfide on the membrane and DNA of tumor cells in culture. A disulfide, n-butyl 2-imidazolyl disulfide (III-2), recently reported to be more toxic to EMT6 tumor cells under hypoxia has also been shown to be preferentially toxic to KHT/iv cells under hypoxia with an IC90 value 3-fold lower versus that measured in air. The IC90 values for both cell lines were markedly affected when the pHe was decreased. Uptake studies revealed that the disulfide is metabolized at the cell membrane with only one of the metabolites, n-butanethiol, being taken up into the cell. There were no differences in the amount of uptake under oxia or hypoxia. Investigation of potential membrane damage revealed that III-2 was a mixed inhibitor of the enzyme Na+,K(+)-ATPase, from isolated plasma membrane. DNA damage, when examined using plasmid DNA in the absence or presence of glutathione, was absent. Minor single-strand breaks to EMT6 DNA could only be observed following exposure to III-2 at the highest concentrations tested, with no differences observed between treatments in air or under hypoxia.


Mechanisms of glucagon-induced increases in rates of cytochrome P450-dependent pentoxyphenoxazone O-depentylation in cultured rat conceptuses. Inclusions of glucagon (1.0 or 2.0 microM, final concentrations) in the media of cultured whole rat conceptuses resulted in concentration-dependent increases in measured rates of O-depentylation of pentoxyphenoxazone in cell-free preparations of conceptal tissues. Enzymic activities were assayed 24 h after initial exposure of the conceptuses to glucagon on day 10 of gestation. Glucagon elicited increases in tissue levels of cAMP that were parallel to those produced by 3-isobutyl-1-methylxanthine over the same time period. Tissue cAMP levels were maximal after 2 h, rapidly returned to control levels and were also equal to background levels in controls after the 24 h culture period. Dibutyryl cAMP, 3-isobutyl-1-methylxanthine, theophylline, and RO201724, a cAMP-selective phosphodiesterase inhibitor, each produced 75 to 100% increases in O-dealkylase activity. Dibutyryl cGMP and two phosphodiesterase inhibitors, enoximone (cGMP-inhibited) and zaprinast (cGMP-specific), each failed to produce statistically significant increases in O-depentylase activity. The O-depentylase was tentatively identified as a conceptus-specific P450 cytochrome that is synthesized predominantly in tissues of the visceral yolk sac. The results indicated that glucagon may upregulate a unique, xenobiotic-biotransforming P450(s) via a long-term mechanism(s) specifically involving tissue cAMP.


[Treatment of climacteric syndrome using Estraderm TTS and premarin in a comparative study]. In an open-label, multicentric randomized trial the efficacy and tolerability of Estraderm TTS, a 17 beta-estradiol, and Premarin, consisting of conjugated estrogens, were compared in the treatment of the climacteric syndrome. 84 patients with manifest menopausal complaints were randomized into two groups of 42 women each. The duration of treatment was 11 weeks or 3 cycles, each with 3 weeks of estrogen treatment followed by a therapy-free week. Therapeutic efficacy was assessed with a questionnaire recording frequency and intensity of hot flushes and of sweating episodes during the night, changes in psychic well-being, frequency of micturition and dryness of the vagina. Systemic and local tolerability was also evaluated. Both substances proved almost equivalent in the treatment of menopausal complaints, although Estraderm TTS was markedly superior in suppressing vasomotor symptoms.


[False rubella diagnosis in pregnancy by IgM enzyme immunoassay]. On the basis of a case report of a false-positive rubella diagnosis in early pregnancy the interpretation of positive IgM is discussed. A primigravida at 14 weeks of gestation showed rubella-specific IgM in an enzyme immunoassay. This result could not be confirmed by an alternative method (hemagglutination-inhibiting test after separating the serum by density gradients into different immunoglobulin classes and removal of the IgG with protein A). The immunological status corresponded with a past rubella infection or with the status after vaccination (IgG-positive). It is therefore recommended to use an additional method in the presence of IgM, especially if an interruption of the pregnancy has to be considered. In the course of the examination a recent cytomegalovirus infection could be detected.


[Alcoholic mitochondrial cardiomyopathy]. The effect of alcohol on oxidative phosphorylation, activity of cytochrome oxidase and the content of cytochromes in mitochondria of the heart muscle was studied in rabbits after 14- and 21-day subcutaneous administration of 20% alcohol solution. Alcohol was found to exert an inhibitory effect on oxygen consumption and on the rate of energy production in mitochondria of the heart muscle, to decrease the specific activity of cytochrome oxidase and to increase the cytochrome content in the mitochondria. The authors classify these metabolic derangements under the concept of alcoholic mitochondrial cardiomyopathy and discuss broader aspects of mitochondrial diseases in medicine.