Gastrointestinal complaints in relation to dietary intake in triathletes. This study examined the relationship between gastrointestinal (GI) symptoms and dietary intake in triathletes. Fifty-five male triathletes (age 31 +/- 6 yrs) were surveyed regarding the most recently completed half Iron Man triathlon. Questions were asked regarding GI symptoms and dietary intake. Fifty-two percent complained of eructation and 48% of flatulence. Other symptoms were abdominal bloating, vomiting urge, vomiting, nausea, stomachache, intestinal cramps, and diarrhea. More symptoms occurred while running than at other times. All individuals who had eaten within 30 min of the start vomited while swimming. Fat and protein intake was greater in those who vomited or had the urge to vomit than in those without these symptoms. Of the former, 93% had consumed a hypertonic beverage. Forty percent of those who drank a hypertonic beverage and only 11% of those who drank an iso- or hypotonic beverage had severe complaints. Four of five individuals with stomachache had consumed a strongly hypertonic beverage. All subjects with intestinal cramps had eaten fiber-rich foods in the prerace meal; only 10% of those without cramps had done so.


[The importance of assessing angiotensin-converting activity in silicosis patients]. The macrophage cells have an essential role in the pathogenesis of silicosis. After the endocytosis of silica grains, the lung macrophage demonstrates an active state, associated with high levels of lysosomal enzymes, expressed also in serum. A similar active state of macrophages is evident in sarcoid lung which manifests also an intensive serum activity of angiotensin-converting enzyme. The angiotensin-converting assay in the serum of 116 silicotic patients revealed over 60 per cent higher levels than in normals. Such increased levels were present more frequently among the patients with early studies of silicosis. In the same patients, the serum acid phosphatase activity, a marker of macrophage activity, and also serum lactate dehydrogenase activity, were higher than in normal individuals. On present evidence, it can be appreciated that increased serum levels of angiotensin-converting activity could be the expression of an active progressive state of silicosis.


Effects of coenzyme athletic performance system as an ergogenic aid on endurance performance to exhaustion. This study examined the effects of the Coenzyme Athletic Performance System (CAPS) on endurance performance to exhaustion. CAPS contains 100 mg coenzyme Q10, 500 mg cytochrome C, 100 mg inosine, and 200 IU vitamin E. Eleven highly trained male triathletes were given three daily doses of either CAPS or placebo (dicalcium phosphate) for two 4-week periods using a double-blind crossover design. A 4-week washout period separated the two treatment periods. An exhaustive performance test, consisting of 90 minutes of running on a treadmill (70% VO2max) followed by cycling (70% VO2max) until exhaustion, was conducted after each treatment period. The mean (+/- SEM) time to exhaustion for the subjects using CAPS (223 +/- 17 min) was not significantly different (p = 0.57) from the placebo trial (215 +/- 9 min). Blood glucose, lactate, and free fatty acid concentrations at exhaustion did not differ between treatments (p < 0.05). CAPS had no apparent benefit on exercise to exhaustion.


[Immunohistochemical observation of angiotensin converting enzyme in acute lung injury]. The morphological changes of acute lung injury of rabbits, particularly the ACE content in the vascular endothelium of lung were studied with immunohistochemical methods and Q-970 image analysor. The activity of ACE in serum collected from the arteries was also estimated. The results showed that during the process of acute lung injury, the lung vascular endothelium was severely damaged with an obvious decrease of ACE content and activity. It indicates that decrease of ACE content and activity may play an important role in the pathogenesis of acute lung injury and may also be the sensitive criteria in expressing damage of the vascular endothelium in the lungs.


[The effect of the pressure of metabolically inert gases and temperature on the membrane fluidity of liposomes]. The comparative analysis of pressure effects of helium, nitrogen (to 100 kg/cm2), cryptone (to 50 kg/cm2) and temperature (32-45 degrees C) on the lateral diffusion of pyrene in liposomal membranes of egg phospholipids was performed. The linear additivity of studied effects was demonstrated. The results are discussed from the point of view of the disturbances of cell processes in the hyperbaric conditions.


[Pharmacological activities of musk. IV. Effects of musk on arachidonic acid metabolism in leukocytes from rat inflammatory exudate]. The present work was designed to further study the mechanism of the anti-inflammatory action of musk from the viewpoint of arachidonic acid (AA) metabolism: 1) effect on AA release; 2) action on leukotriene B4 (LTB4) and 5-HETE biosyntheses. Leukocytes obtained from rat inflammatory exudate by i.p. injection of 1% carrageenan were labelled with 14C-AA at 37 degrees C for 2 h. The 14C-AA labelled cells were incubated with the water soluble fraction of musk (MWF) and stimulated with calcium ionophore A23187 to release AA. The radioactivity released from the labelled leukocytes was determined as a criterion of AA release. In order to observe the influence of musk on the biosyntheses of LTB4 and 5-HETE, leukocytes were incubated with MWF and AA, and then stimulated with A23187. The LTB4 and 5-HETE produced were determined by RP-HPLC. The results showed that AA release decreased to 50-70% of the control at MWF concentration of 6.25-37.5 micrograms/ml. The biosyntheses of LTB4 and 5-HETE were not affected. However, AA release increased to 1.5-fold at the MWF concentration of 400 micrograms/ml. At the same time, biosyntheses of LTB4 and 5-HETE decreased by 66.7 and 90%, respectively, as compared with controls. This result indicates that MWF not only inhibited phospholipase A2 (PLA2) activity, but also prevented AA peroxidation and inhibited LTB4 biosynthesis. These results suggest that inhibition of the AA metabolic pathway may play a major role in the mechanism of the anti-inflammatory action of musk.


[The physicochemical properties of hydrogen and its use as an indifferent component in breathing mixtures under high pressure]. Data on a possible use of hydrogen as a part of the breathing mixtures during deep diving are presented; advantages and disadvantages of hydrogen application in the undersea research and diving practice are evaluated. Perspectives and trends with respect to a safe hydrogen use under high pressure in combination with oxygen and other inert gases are discussed.


Ultracytochemical studies on Trichomonas hominis. The results of ultracytochemical studies on Trichomonas hominis showed that ACPase and CMPase were mainly located in the mature face sacs of the primary lysosomes, digestive vacuoles, as well as in the parabasal body. TPPase and NADPase were found in the saccules at the mature face and the intermediate saccules of parabasal body respectively. This study revealed that T. hominis had well-developed parabasal bodies. Negative COase and catalase reactions indicated that T. hominis lacked both mitochondria and microbody. Hydrogenosome was stained well with the Ur-Pb-Cu impregnation technique.


CT-guided stereotactic injection of radionuclide in treatment of brain tumors. 140 patients with brain tumor were treated by CT-guided stereotactic injections of radionuclides, such as Aurum-198 (198 Au), Phosphorus-32 (32 P) and Yttrium-90 (90Y). Of these patients aged from 3 to 67 years (average 37), 64 were male and 76 female. Astrocytoma was found in 75 patients, craniopharyngioma in 46, metastatic carcinoma in 7, meningioma in 5, germinoma in 4 and pituitary adenoma in 3. The tumors were located in the deep part or functionally critical area of the brain. After 267 times of injection of colloidal isotopes, no major adverse effects or complications occurred. Follow-up for 6 to 48 months showed improvement in symptoms in 104 (74.3%) patients and CT scanning showed the diminished tumors.


[Ocular findings in pituitary adenoma].

[Ocular findings in pituitary adenoma]. Of 315 cases (629 eyes) of surgically confirmed pituitary adenoma, 487 eyes (77.4%) suffered diminished visual acuity, 393 eyes (62.5%) primary optic atrophy and 500 eyes (79.5%) visual field defects. Ophthalmoplegia and papilledema were also observed. The ocular changes and visual prognosis were discussed.


In-vitro activity of daptomycin against a worldwide collection of methicillin-resistant Staphylococcus aureus. Minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of daptomycin and of vancomycin have been compared against 80 strains of methicillin-resistant Staphylococcus aureus isolated from many parts of the world. In the presence of 30 mg/l of Ca++, daptomycin at 4 mg/l killed all the strains tested, and was only slightly less active than vancomycin.


A comparative study on the convulsant activity of carbapenems and beta-lactams. Beta-lactams are well known to have potent convulsant activity. Imipenem, the first carbapenem antibiotic introduced in the clinical field, has been reported to induce convulsions. The neurochemical mechanism of the convulsions induced by carbapenems and cephalosporins were studied. Intraventricular injection of cefazolin, cephaloridine and imipenem, and of panipenem (a new carbapenem), induced convulsions in a dose-dependent manner in mice. They inhibited the receptor binding of gamma-aminobutyric acid (GABA), an inhibitory transmitter in the mammalian central nervous system. These in-vitro and in-vivo results suggest that carbapenems and cephalosporins might induce convulsions through the inhibition of GABA receptor binding when they are accumulated in the central nervous system.


Metabolic effects of rectal administration of unmodified eel calcitonin in humans. To evaluate the metabolic effects of unmodified eel calcitonin, nine normal subjects and eleven patients with Paget's disease of the bone entered the study. Eel calcitonin was administered via rectal mucosa at the dosage of 100 and 200 MRCU. Plasma calcium, plasma phosphate and plasma cAMP were measured at a baseline and after 10, 20, 30, 60, 90, 120, 180, 240 min. Rectal administration of 100 MRCU of eel calcitonin decreased plasma calcium and phosphate levels in normal subjects; the hypocalcaemic and hypophosphatemic effects were more marked and statistically significant in Paget's disease patients. Using 200 MRCU of eel calcitonin, statistically significant reductions in plasma calcium levels were observed in controls and in Paget's disease patients. Plasma phosphate decreased after rectal administration of the hormone, but not significantly. A slight but not significant increase in plasma cAMP was observed in normal subjects after the administration of 200 MRCU. These data demonstrate that eel calcitonin administered via rectal mucosa can exert some of the known biologic effects of the peptide.


Detection of a near infra-red absorption band of ferrohaem a3 in cytochrome c oxidase. We have detected weak absorption bands in the near infra-red region of reduced mammalian cytochrome c oxidase, analogous to those that we have recently reported to be present in the bacterial cytochrome o (Ingledew, W.J., Bacon, M. and Rich, P.R. (1992) FEBS Lett. 305, 167-170). The major band is centred at 784 nm and has an sigma mM-1.cm-1 of around 0.1. It is shifted to 760 nm in the carbon monoxide compound and is absent in the reduced cyanide complex. We attribute it to a charge transfer band of ferrohaem a3, equivalent to the 'band III' or 'conformational band' of haemoglobin.


Electron spin resonance study on the permeability of superoxide radicals in lipid bilayers and biological membranes. The permeability of lipid bilayers and biological membranes to superoxide free radicals was examined by using superoxide dismutase (SOD)-loaded lipid vesicles and SOD-loaded erythrocyte ghosts. After exposing SOD lipid vesicles and SOD ghosts to enzymatically produced superoxide radicals and using spin-trapping and electron spin resonance (ESR) techniques, we found that SOD entrapped within erythrocyte ghosts effectively scavenges external O2.- while SOD inside the lipid bilayers has no effect. These results confirm that O2.- is able to cross through a biological plasma membrane but not across a pure lipid bilayer. The data provide instruction as to how and where anti-oxidant therapy is to be approached relative to the site of oxygen free radical production.


Activation of nucleoside diphosphate kinase by mastoparan, a peptide isolated from wasp venom. We have previously reported that GDP-bound alpha beta gamma-trimeric GTP-binding (G) proteins can be converted into the active GTP-bound form with nucleoside diphosphate (NDP) kinase and ATP, although its exact activation mechanism still remains to be resolved. In the present study, we investigated whether NDP kinase activity was modified by mastoparan, a wasp venom peptide that is known to activate G proteins as an agonist-receptor complex. The activity of NDP kinase measured by the formation of GTP from ATP and GDP was markedly stimulated, when the kinase was incubated with mastoparan. The concentration of mastoparan required for the activation was much lower than that observed for the peptide-induced activation of G proteins under similar assay conditions. There was also an increase in the phosphorylated intermediate of NDP kinase as well as the catalytic activity upon its incubation with mastoparan. These results suggest that mastoparan not only activates G proteins directly via guanine nucleotide exchange reaction but also stimulates NDP kinase activity.


Bactericidal activities of single or multiple doses of various combinations of new antileprosy drugs and/or rifampin against M. leprae in mice. The bactericidal activities against Mycobacterium leprae of single or multiple doses of various combinations of new antileprosy drugs [minocycline (MINO), clarithromycin (CLARI), ofloxacin (OFLO), and sparfloxacin (SPFX)] and/or rifampin (RMP) were titrated in immunocompetent mice by the proportional bactericidal method. Drugs were administered by gavage at the following dosages (mg/kg) per dose: RMP 10, MINO 25, CLARI 100, OFLO 150, and SPFX 50. All 15 regimens exerted significant bactericidal activities, at least 96% of viables were killed. The activity of a single dose MINO + CLARI was only slightly inferior to that of RMP, and the activities of a single dose OFLO/SPFX + MINO + CLARI were similar to that of RMP. This suggests that either MINO + CLARI or OFLO/SPFX + MINO + CLARI may be administered once monthly together with RMP 600 mg for the treatment of multibacillary (MB) leprosy, and monthly administration of MINO + CLARI or OFLO/SPFX + MINO + CLARI may also be employed for the treatment of RMP-resistant MB leprosy. Because the killing effects of multiple doses of the combinations were so powerful, comparison of the bactericidal activities of these regimens was beyond the sensitivity of the immunocompetent mouse model, and are being tested in the nude mouse model. Although SPFX is more active against M. leprae than OFLO on a weight-to-weight basis, when both drugs were administered in mice at dosages equivalent to clinically tolerated dosages in humans, SPFX did not show more superiority than OFLO, and its real advantage over OFLO in the treatment of leprosy remains unclear.


Effects of dietary magnesium supplementation on diurnal variations of blood pressure and plasma Na+, K(+)-ATPase activity in essential hypertension. To evaluate the antihypertensive and hormonal effects of oral magnesium supplementation, 17 inpatients with untreated, uncomplicated mild-to-moderate essential hypertension (EH) and 8 age-matched normotensive controls (controls) were given MgO orally 3 times a day at a daily dose of 1.0 g (0.6 g per day as Mg) for a period of 2 weeks. Supplementation of MgO elicited a significant fall in averaged mean blood pressure calculated with a 24-h ambulatory blood pressure monitoring system (MBP) in EH from a baseline value of 104.3 +/- 12.2 to 99.5 +/- 11.6 mmHg (p < 0.05), while controls remained unaltered from a baseline value of 85.1 +/- 11.5 to 84.5 +/- 13.3 mmHg. The percentage reductions in systolic and diastolic blood pressures were similar during daytime and nighttime in EH. According to the extent of reduction in MBP with magnesium supplementation, EH patients were divided into 2 groups, responder and nonresponder. The level of plasma renin activity (PRA) in the responder group was significantly higher than that of the nonresponder group (p < 0.05). After 2 weeks of magnesium supplementation, the plasma level of Na+, K(+)-ATPase inhibitory activity (PATPI), defined as equivalency to ouabain, was reduced significantly from 0.75 +/- 0.54 to 0.40 +/- 0.30 mumol ouabain/ml (p < 0.05) in the responder group, while it remained unaltered in controls and the nonresponder group. PRA, plasma aldosterone concentration, urinary epinephrine and norepinephrine excretion, and urinary sodium excretion did not change significantly in either control subjects or EH (responder and nonresponder groups). A significant negative correlation existed between the pretreatment PRA and changes in MBP after magnesium supplementation in EH (r = -0.65, p < 0.01), and there was a significant positive correlation between changes in PATPI and changes in MBP as a whole (r = 0.41, p < 0.05). These results support the view that oral magnesium supplementation is a useful approach to treatment of patients with uncomplicated essential hypertension, especially those with high plasmas renin activity. It appears that magnesium suppresses circulating Na+,K(+)-ATPase inhibitory activity to attenuate vascular tone, and thereby reduces blood pressure in EH.


[Anti-polynuclear neutrophil cytoplasmic antibodies of the perinuclear type or p-ANCA]. Some neutrophil cytoplasmic auto-antibodies produce perinuclear immuno staining of alcohol-fixed neutrophils called P-Anca pattern. These autoantibodies show chiefly reactivity with myeloperoxidase in Elisa but other specificities have been detected (elastase, cathepsin, lactoferrin). These P-Anca anti-MPO are more frequent with renal angiitis but P-Anca (anti-MPO negative) are associated with other diseases without renal failure (rheumatoid arthritis, Gougerot-Sjogren and ulceritis colitis).


Avidity and titer of immunoglobulin G subclasses to Porphyromonas gingivalis in adult periodontitis patients. The relative avidity and titer of antibodies representing the 4 immunoglobulin G (IgG) subclasses (IgG1-4) reactive with Porphyromonas gingivalis, P. gingivalis-lipopolysaccharide (-LPS), streptokinase (SK) and tetanus toxoid (TT) in the sera of patients having adult periodontitis and of healthy controls were measured. Patient antibody titers to P. gingivalis and P. gingivalis-LPS were found to be significantly elevated for IgG, IgG1 (no P. gingivalis-LPS antibodies) and IgG2. The predominant antibody response to P. gingivalis and P. gingivalis-LPS occurred in the IgG2 subclass. When the relative avidity of the antibodies to P. gingivalis and P. gingivalis-LPS were examined, no significant differences between control and patient sera could be identified. However, anti-P. gingivalis and P. gingivalis-LPS antibodies were found to possess significantly lower relative avidity than either SK or TT antibodies. The IgG1 subclass antibodies to P. gingivalis, SK and TT all appeared to be of high relative avidity. In contrast, anti-P. gingivalis and P. gingivalis-LPS of the IgG2 subclass were of significantly lower relative avidity. Since the predominant humoral response to P. gingivalis occurs in the IgG2 subclass, the low relative avidity of these antibodies predominates in measurements of whole serum activity.


Evaluation of a non-radioactive DNA probe for detecting Porphyromonas gingivalis in subgingival specimens. This study compared the ability of a nonradioactive digoxigenin-labeled DNA probe and anaerobic culture to identify subgingival Porphyromonas gingivalis. Total cellular DNA from P. gingivalis ATCC 33277T was labeled using the Genius kit from Boehringer Mannheim Biochemicals. Anaerobic culture was performed using VMGA III transport medium and enriched brucella blood agar. The DNA probe could detect as little as 1000 P. gingivalis cells added to supragingival plaque. Also, the probe could detect P. gingivalis when it was present in proportions too low to be visualized on overgrown bacterial plates. The probe showed no visible reaction with strains of various oral species or with thousands of non-P. gingivalis colonies from plaque samples. VMGA III could maintain the viability of P. gingivalis for up to 6 days, as evidenced by DNA probing of colony blot of subgingival cultures. A total cellular DNA probe for detecting P. gingivalis seems to offer a simple and reliable method of detecting the organism in subgingival specimens.


Expression of Porphyromonas gingivalis proteolytic activity in Escherichia coli. Porphyromonas gingivalis (formerly Bacteroides gingivalis) degrades numerous protein substrates including collagen, fibrinogen, fibronectin, gelatin, casein, immunoglobulins and complement components. In order to clone one or more of these protease genes, a genomic library was constructed with Sau3A1 restriction fragments of chromosomal DNA from P. gingivalis ATCC 33277 ligated into the temperature-regulated vector pCQV2, and expressed in Escherichia coli DH5 alpha mcr. The electro-transformants (3 x 10(4)) were screened for general protease activity on Luria broth agar containing ampicillin (50 mg/l) and sodium caseinate (2%). One casein-hydrolyzing clone was detected and subcultured, and the activity of the cell extracts was characterized. We were able to show that the protease-positive clone, (pTEM1), had broad substrate specificity. Colorimetric assays indicated the hydrolysis of azocoll, azocasein, collagen, elastin-congo red and artificial substrates. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to confirm that collagen, casein, fibrinogen and fibronectin were degraded by the clone.


Interaction of Porphyromonas gingivalis with gingival epithelial cells maintained in culture. Interactions between Porphyromonas gingivalis and gingival epithelial cells were investigated. Gingival epithelial cells were cultured from surgically removed gingival tissue. Electron microscopy demonstrated adherence of P. gingivalis to the cell membranes and microvilli followed by internalization of the bacteria into the epithelial cell cytoplasm. Saliva from healthy and periodontally diseased patients inhibited P. gingivalis association with the epithelial cells. Attachment to and penetration of gingival epithelial cells by P. gingivalis may be important virulence factors in periodontal disease. Salivary molecules may play a role in modulating these interactions.


Identity of amino acid sequences of superoxide dismutase purified from both anaerobically maintained and aerated Porphyromonas gingivalis. Superoxide dismutase (SOD) from anaerobically maintained Porphyromonas gingivalis (anaero-SOD) has the characteristic of Fe-SOD, whereas SOD from aerated cells (aero-SOD) has that of Mn-SOD. However, both types of apoSOD can bind either Fe or Mn. To elucidate the structure relationship between anaero- and aero-SOD, we examined the amino acid sequence of aero-SOD by direct protein analysis. The amino acid sequence of aero-SOD was shown to be identical with that of anaero-SOD determined previously. Our findings support the hypothesis that cambialistic SOD is formed from a single apoprotein in bacterial cells.


Antibodies, implantation and embryo survival. The diverse strategies adopted among species for the maintenance of luteal function converge to meet the indispensable requirement for this hormone particularly at the time of onset of implantation. What has emerged from immunization studies is the difference that exists between various species in the effects of progesterone depletion. The findings affirm the uterus as a primary site of progesterone action in the preimplantation period of gestation in the mouse, whereas in the hamster the positive feedback by progesterone on pituitary luteotrophin secretion is important in maintaining luteal function and hormone secretion at a level necessary for uterine preparation. The impact of progesterone in the rat seems to be established within 48 h after fertilization, whereas in the ferret the hormone's action ensures a suitable uterine environment in which the early embryo can flourish. Its effects in early pregnancy in the marmoset are ambiguous from the present studies unless it emerges that target organ responses are tuned to low concentrations of active steroid. However, the discoveries in mice of a conserved family of immunoglobulin genes used exclusively by immunogenic forms of progesterone conjugated to proteins to stimulate antibody production, and of antibody binding to the uterine epithelium, reveal systems potentially inimical for embryo survival. The endometrial expression of the proto-oncogene erb-A at a time when the embryo has not yet arrived in the uterus, and of antibody-binding to the uterine epithelium, are early maternal responses whose significance require closer examination. The present findings support the hypothesis that the primary site of action of progesterone during the preimplantation period differs between species, and that it is not only the mother that recognizes, but the embryo that initiates early changes before the onset of implantation 'in the bed of soil that nourishes it', a symbiosis that may yet prove to be a common feature of the adoption of viviparity as a preferred mode of reproduction.


Effect of continuous infusion of oxytocin on ovarian function and uterine oxytocin receptor concentration in the cyclic ewe. Intact cyclic ewes have been used in experiments designed to examine the mechanism by which uterine oxytocin receptor synthesis is controlled during the oestrous cyclic. Previous experiments have shown that the prostaglandin F2 alpha analogue cloprostenol is luteolytic in ewes receiving oxytocin by continuous intra-venous infusion. When ewes receiving oxytocin are given cloprostenol uterine oxytocin receptor concentrations are raised, whereas in animals receiving oxytocin alone, they remain low. To investigate whether inhibition of oxytocin receptor binding activity by oxytocin is either dependent on elevated plasma progesterone concentrations or over-ridden by oestrogens secreted by ovarian follicles maturing as a result of cloprostenol treatment, ewes were given oxytocin by continuous intravenous infusion (3 nmol h-1) between Days 12 and 17 after oestrus and one of the following: no further treatment; cloprostenol [125 micrograms intramuscularly (i.m.)] on Day 15; progesterone, by subcutaneous implant, from Day 14 with cloprostenol on Day 15; medroxyprogesterone acetate (MPA; 6 mg depot i.m.) on Day 14 followed by cloprostenol on Day 15; or oestradiol-17 beta (2.75 mumol i.m.) on Days 14 and 15. Concentrations of oxytocin receptor were measured at autopsy on Day 17 in caruncular endometrium, intercaruncular endometrium and myometrium. Ovarian follicles and corpora lutea were examined to determine the effect of treatment on these tissues. Treatment with oxytocin alone resulted in the maintenance of corpora lutea, reduced follicular development and a low concentration of uterine oxytocin receptor. Cloprostenol initiated luteolysis in oxytocin-treated ewes. This was associated with a high level of oxytocin receptor binding activity in all ewes except those receiving exogenous progesterone.(ABSTRACT TRUNCATED AT 250 WORDS)


[What have we learned from converting enzyme inhibitors on renin-angiotensin system?]. Angiotensin-converting-enzyme (ACE) inhibitors are now widely used to treat patients with high blood pressure or heart failure. The favourable results obtained with these inhibitors of the renin-angiotensin system suggest that angiotensin II has a noxious effect on the development and/or course of these diseases. ACE inhibitors are usually well tolerated. Their most severe side-effects are mostly foreseeable and therefore avoidable. Chronic blockade of the renin-angiotensin system increasingly seems to be a good therapeutic approach to the protection of the vital organs.


[Calcium antagonism of anisodamine on isolated rabbit thoracic aorta]. Anisodamine (Ani) or verapamil (Ver) relaxed the contracted thoracic aortic strips induced by CaCl2 and KCl in a noncompetitive manner with pD2' 3.6 and 5.8, respectively. Ani 50 mumol.L-1 and Ver 1 mumol.L-1 inhibited the intracellular Ca(2+)-dependent component of norepinephrine-induced contraction of aorta, but Ani 0.1 mmol.L-1 did not decrease the extracellular Ca(2+)-dependent component. In the presence of Ca2+, Ani 2.3 mumol.L-1 inhibited the proliferation of aortic smooth muscle cells. In the absence of Ca2+, the effect induced by Ani was biphasic-stimulatory at 2.3 mumol.L-1 concentrations and inhibitory at 61.7 mumol.L-1 concentrations. These results suggest that Ani, similar to Ver, mainly inhibits the potentially operated calcium channels.


[Effects of spermine on phosphorylation of phosphoinositide and 47 K protein in platelet membranes of pig]. In the presence of Mg2+ and exogenous phosphatidylinositol-4-phosphate, swine thrombocytic membranes were incubated with [gamma-32P]ATP at 30 degrees C for 3 min and the incorporations of 32P into phospholipids and proteins were measured. Spermine stimulated the formation of phosphatidylinositol-4,5-bisphosphate (optimal concentration 0.25 mmol.L-1) and inhibited the phosphorylation of 47 K protein (IC50 1 mmol.L-1). This helps to clarify the effect of spermine on cell growth and proliferation.


Molecular bioengineering of biomaterials in the 1990s and beyond: a growing liaison of polymers with molecular biology. An important trend in biomaterials research and development is the synthesis of polymers that combine capabilities of biologic recognition (biomimetic) with special physicochemical properties of the synthetic polymer system. Another important trend in such "molecular bioengineering" is to develop, perhaps via computer-aided molecular design, new artificial biomimetic systems by exact placement of functional groups on rigid polymer backbones, cross-linked structures, or macromolecular assemblies. In this way, biocatalytic functioning or biorecognition similar to enzymes and antibodies can be achieved without the inherent instability often encountered with the native biomolecules or assemblies. Perhaps the most exciting trend in biomaterials research and development is the availability of new biomolecules, e.g., via protein engineering and of hardy cells with specific biofunctions and bioresponses that can be tailored to specific medical or biotechnological needs. The wide variety of ways that such biomolecules and cells can be combined with polymeric biomaterials provides tremendously exciting opportunities for the biomaterials scientists and engineers. In addition to these synthetic approaches, new and exciting analytical tools, such as the scanning tunneling microscope and the atomic force microscope, are permitting study on a molecular scale of individual and small clusters of proteins and other biomolecular assemblies on surfaces. Cell attachments and spreading may also be visualized at various depths within the cell using the confocal laser microscope. Such analytical techniques can lead to important new knowledge about biologic interactions with biomaterials and, therefore, to development of even more biocompatible implants and devices. This paper overviews the present state of polymeric biomaterials and highlights the important and exciting opportunities generated by the liaison of these materials with molecular biology.


Bioartificial organs.

Bioartificial organs. Bioartificial organs combine the physical aspects of implantable prostheses with the biological advantages of organ transplantation. Enclosing live cells in a permselective, synthetic envelope avoids rejection by an immunoincompatible host while the geometric limitation of the closed polymer capsule prevents overgrowth of the transplanted material. As a tenet bioartificial organs widen the range of therapeutics based on the biological activity of cell transplants and open an alternative path to gene therapy.


Artificial cells: 35 years.

Artificial cells: 35 years. The first artificial cells were prepared 35 years ago. They contain biologically active materials. They are now being used in medicine and biotechnology. Artificial cells containing adsorbents are already a routine form of treatment in hemoperfusion. This includes treatment for acute poisoning, high blood aluminum and iron, kidney failure, some types of acute liver failure, and other conditions. Artificial cells are being tested for use as red blood cell substitutes. Artificial cells containing cell culture are being tested in animals for the treatment of diabetes, liver failure, and others. Artificial cells containing enzymes are being tested for treatment in hereditary enzyme deficiency diseases and other diseases. Artificial cells containing complex enzyme system can convert wastes like urea and ammonia into useful amino acids. In biotechnology, artificial cells are being used for the production of monoclonal antibodies, interferons, and other biotechnological products. They are also being investigated for use in other applications in biotechnology, chemical engineering, and medicine.


Positive lusitropic effect and diminished myofibrillar sensitivity to calcium produced by cAMP on toad (Bufo arenarum Hensel) ventricle. In intact ventricular strips from toad heart, we studied the relaxant or positive lusitropic effect of different interventions known to increase intracellular cAMP levels. Isoproterenol increased developed tension (DT), maximal rate of contraction (+T), and maximal velocity of relaxation (-T). From 10(-8) to 10(-4)M isoproterenol, -T increased proportionally more than +T being the ratio +T/-T significantly decreased. A single dose of isoproterenol (3 x 10(-8)M) increased cAMP levels from 0.174 +/- 0.022 to 0.329 +/- 0.039 pmoles/mg ww (P < 0.05), increased contractility by 69 +/- 13% and decreased +T/-T by 18.5 +/- 4.55%. Administration of 10(-3)M of dibutyryl cyclic AMP (dcAMP) significantly increased DT and +T and decreased the ratio +T/-T. Similar effects were obtained with milrinone, a specific cAMP phosphodiesterase inhibitor. Papaverine, a non selective phosphodiesterase inhibitor, failed to increase +T but significantly increased -T. In chemically skinned ventricular trabeculae, calcium sensitivity of the myofibrils was significantly increased by 10(-5)M of the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX). 10(-3)M dcAMP failed to affect calcium sensitivity of chemically skinned ventricular trabeculae when given alone, but produced a decrease in calcium sensitivity of the myofibrils in the presence of 10(-5)M of either IBMX or papaverine. The results would indicate that the relaxant effect of isoproterenol is mediated in toad ventricle by an increase in intracellular cAMP levels. They furthermore suggest that a decrease in myofilament sensitivity to calcium may be a mechanism by which cAMP produces its relaxant effect.


Is the renal (Na + K)-ATPase modulated by intracellular messengers? The basic cellular mechanisms involved in the regulation of (Na + K)-ATPase are discussed. Various ligands seem to be responsible for the short-term modulation of this enzyme activity (intracellular messengers). Cytosolic Ca2+ has a key role in mediating changes induced by hormones or receptor agonist; but, in turn, intracellular Ca(2+)-dependent proteins like calmodulin, calnaktin or others, are also needed for these changes. Phosphorylation of effector proteins, following the activation of PKC, PKA or CaM-kinase II, may result in changes of (Na + K)-ATPase activity either by a direct effect on the catalytic subunit or by modulating the Na(+)-H+ exchanger thereby resulting in an effect on intracellular sodium, whose concentration is known to be rate-limiting for the enzyme activity. Despite the ubiquity of (Na + K)-ATPase in various organs and tissues, its response to modulators partly depends on the heterogeneity of the alpha-subunit that give rise to the existence of different isoforms. The relative abundance of alpha 1, alpha 2, alpha 3 or other isoforms is tissue-specific and represents another way of regulation among different cell types. While these cellular mechanisms occur in various cell types the kidney shows an opposite response respect to other tissues such as liver or brain. The functional relevance of the mechanisms of acute adaptation of (Na + K)-ATPase, discussed in this review, is becoming increasingly recognized for the renal enzyme, what may contribute to stimulate new approaches to the study of the short-term regulation of the pump activity in molecular terms.


Cytomegalic inclusion disease presenting acute intrahepatic cholestasis. An 83-year-old man suffering from pulmonary emphysema was admitted to our hospital because of jaundice. He was diagnosed as acute intrahepatic cholestasis but the etiology could not be determined during the treatment period. In spite of treatment, the jaundice worsened progressively without any elevation in serum transaminase, and he died of respiratory failure 58 days later. An autopsy revealed a generalized cytomegalic inclusion disease, predominantly in the biliary tracts, liver and lungs. This is a rare case of cytomegalic inclusion disease presenting acute intrahepatic cholestasis without any elevation of transaminase during the clinical course.


An adult case of severe chronic active Epstein-Barr virus infection syndrome. A 23-year-old man with persisting high fever developed hepatosplenomegaly, lymphadenopathy and massive pericardial effusion. Immunological examination revealed a marked elevation of anti-Epstein-Barr virus antibodies (anti-viral capsid antigens IgG-antibody 1:10,240, anti-early antigens-DR IgG-antibody 1:5,120), decreased activities of Epstein-Barr virus specific cytotoxic T lymphocytes, natural killer cells and lymphokine activated killer cells. A liver biopsy showed moderate sinusoidal lymphocytosis with punched-out lesions. These findings suggested severe chronic active Epstein-Barr virus infection syndrome. The patient was treated with recombinant human interleukin-2, but it was discontinued because of an adverse reaction. Twelve months later, he died of suspected pulmonary infection.


Parallel distributed processing and neuropsychology: a neural network model of Wisconsin Card Sorting and verbal fluency. Neural networks can be used as a tool in the explanation of neuropsychological data. Using the Hebbian Learning Rule and other such principles as competition and modifiable interlevel feedback, researchers have successfully modeled a widely used neuropsychological test, the Wisconsin Card Sorting Test. One of these models is reviewed here and extended to a qualitative analysis of how verbal fluency might be modeled, which demonstrates the importance of accounting for the attentional components of both tests. Difficulties remain in programming sequential cognitive processes within a parallel distributed processing (PDP) framework and integrating exceedingly complex neuropsychological tests such as Proverbs. PDP neural network methodology offers neuropsychologists co-validation procedures within narrowly defined areas of reliability and validity.


[Treatment of experimental liver tumors by in vivo suicide gene transfer in rats]. We investigated whether the transduction of a suicide gene might induce the elimination of malignant solid tumours. BDIX male rats were given an intra-hepatic injection of a mixture containing DHDK12 tumor cells and xenogeneic fibroblasts. The latter were producing either the herpes simplex virus type 1 thymidine kinase HSV1-TK- or nls-lac Z(control)-expressing recombinant retroviral particles. 5 days later, a time at which the tumor is macroscopic, all the rats were treated with ganciclovir, a nucleoside analog that is metabolized by HSV1-TK into a toxic compound. After 5 days of treatment, a dramatic reduction in tumour volume was noted in the HSV1-TK group. These results delineate a new therapeutical strategy for the treatment of disseminated liver metastases or of a large variety of solid malignant tumours.


[Quench effect of metallothionein on free radical signal in post-ischemic reperfusion of isolated rat hearts]. Signals of N-tert-butyl-phenyl-nitrone (PBN) spin adducts were observed during post-ischemic reperfusion in isolated rat hearts using electron spin resonance (ESR) spectroscopy and ESR spin-trap PBN. The intensity of signals was related to the time course of ischemia. The intensity of the signals produced during reperfusion after 40 minutes of ischemia was remarkably higher than that of 20 minutes of ischemia. Metallothionein (MT) did quench the adduct signals and the extent of quenching of the signals by this protein was dose-dependent. The signal intensity was decreased to 40% of corresponding control at the concentration of 5 x 10(-6) mol/L. It is a kind of endogenetic protein rich in cysteine residue and divalent metal ions. It is believed to be a promising cytoprotective drug in clinical medicine.


[Adenovirus types 11 and 21: genome types and clinic features]. DNA restriction endonuclease analysis was made on 26 adenovirus types 11 and 21 (Adv 11, Adv 21)--uncommon adenovirus causing infantile pneumonia with the enzymes BamHI and HindIII in Changchun area from 1975 to 1982. Adv 11 and Adv 21 represented the two new genome types during their prevalence for 8 years, i.e. D2 (11), D3(11), D2(21) and D3(21). The decisive factors of viral virulence and pathogenicity were analysed by comparison of the clinical features of pneumonia caused by different types Adv. The study of DNA structures revealed the similarities and differences in the clinical features of the disease caused by the serotypes and genome types Adv.


[A comparison of the effectiveness of mass immunization campaign of trivalent oral poliovirus vaccine (TOPV) by epidemiological serosurvey. Shandong Collaborating Group for Poliomyelitis Control and Surveillance, Jinan, Shandong]. In order to evaluate the effect of TOPV mass immunization campaign, 902 serum samples from two counties of Shandong province were assayed by micro-neutralization test for the neutralization antibodies against poliovirus. The results showed that among children of 0 to 9 year-old groups, the antibody-positive rates and geometric mean titres (GMTs) with poliovirus type 1 to 3 after mass immunization campaign were significantly higher than those before the campaign, respectively. The author considers that it is necessary for poliomyelitis eradication to carry out TOPV mass immunization campaign on the basis of EPI routine immunization.


[A case of distal myopathy with rimmed vacuole formation, progresses into proximal-dominant muscle involvement]. A case of 55-year-old male with distal myopathy with rimmed vacuole formation is reported. He first noticed dragging of his legs at the age of forty-three. Two years later, he was evaluated to have muscle wasting and weakness in lower legs. In another ten years, he became unable to stand or walk unaided. On physical examination, proximal limb muscles were more severely affected than distal limb muscles. Notably, muscle strength of the quadriceps femoris muscles were weak (MRC Scale 3/5), compared to hamstrings, tibialis anterior muscle and gastrocnemius muscle (4/5). Serum creatine kinase, electromyography, nerve conduction velocities were all compatible with this diagnosis. A computed tomography of the musculoskeletal system was consistent with physical findings. Muscle biopsy revealed many fibers with typical rimmed vacuoles (approximately 6% of fibers). Additionally, small amount of ragged-red fibers (0.5%) was noted. Histochemical reaction showed a focal deficiency of cytochrome c oxidase. This case suggests that during the longstanding course of the illness, proximal limb muscles may be more severely affected, and quadriceps femoris muscle may be predominantly involved.


[Investigation of infection with human papillomavirus in areas of high and low incidences of cervical carcinoma in Sichuan Province]. From Jan. to Oct. 1989, we investigated the role of various life style and dietary factors in areas of high and low incidences of cervical carcinoma, as well as infection with types 16 and 33 of human papillomavirus (HPV) in populations. The polymerase chain reaction (PCR) technique was used to detect the infection with HPV 16/33. The result showed that the infection rate of HPV was about 17.9% (Guangyuan, area of high incidence of cervical cancer), and the infection rate about 3.4% (Mian Zhu, area of low incidence of cervical cancer). It was clear that Guangyuan was 5-fold as the infection rate of HPV 16/33 as Mian Zhu. Strong and statistically significant associations were found to between infection rate of Guangyuan and Mian Zhu (P < 0.001). Other risk factors of cervical carcinoma, such as cigarette smoker, Vegetable intake, genital hygiene, personal hygiene etc, were also discussed and analysed in the article.


[Effects of cytomegalovirus infection in pregnant women to fetuses: study with DNA-DNA hybridization method]. DNA of cytomegalovirus (CMV) was examined in 131 placentae and 28 umbilical blood specimens by DNA-DNA hybridization. The result revealed that CMV DNA was detected in 4 of 50 placentae from abnormal fetuses (31 fetal deaths, 19 fetal deformities). 77 placentae from normal fetuses showed negative results. One of 2 cord blood samples from fetal deformities showed CMV DNA positive. 25 umbilical blood samples from normal term newborns showed negative. 4 placentae and 1 cord blood sample from premature infants showed negative results. The results indicate that CMV may play a great role in fetal death and fetal deformity through the infected maternal-fetal circulation.


Unusual presentation of lacrimal gland tumours. The diagnosis of orbital tumour is a challenge to the ophthalmologist. The aim of this paper is to highlight the unusual presentation of lacrimal gland tumours.


Actions of parathyroid hormone and parathyroid hormone-related protein. By interacting with a structurally identical receptor, parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP) display a common spectrum of action on the transport of mineral elements in bone and kidney. In vivo, PTH/PTHrP similarly reduce the renal tubular reabsorption of inorganic phosphate (Pi) and increase that of calcium. The hypercalcemic effect of PTHrP is due to an increase in both bone resorption and renal calcium reabsorption, the latter through a sodium-independent mechanism. The PTHrP-stimulated bone resorption can be totally inhibited by bisphosphonate therapy. Despite that, the fall in calcemia is moderate, indicating that the PTHrP main hypercalcemic action is due to the stimulation of the renal transport of calcium. For identical effects on renal ionic transports, PTHrP appears to less stimulate bone formation than PTH. These experimental findings are similar to clinical observations in patients with primary hyperparathyroidism or with solid malignant tumors. In vitro, the effects of PTH(1-34), PTHrP(1-34) and PTHrP(1-141) on cAMP production and sodium-dependent phosphate transport (NaPiT) are similar in kidney cells, where NaPiT is specifically inhibited by either peptide. This effect is attenuated by the competitive inhibitor [D-Trp12,Tyr34]bPTH(7-34)amide. Transforming growth factor-alpha similarly modulates the cAMP and NaPiT responses to PTH/PTHrP. In cultured mammary cells isolated from lactating rats, PTHrP elicits a 2-fold increase of cAMP production. Various products of bone and stromal cells, and of leukocytes, such as Interleukin-6 or Tumor necrosis factor-alpha, as well as high extracellular calcium concentration enhance PTHrP production by cultured lung squamous cell carcinoma and Leydig tumor cells, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Aniridia-Wilms' tumour syndrome--a case report. Wilms' tumour is rarely associated with sporadic non-familial congenital aniridia. A child with sporadic aniridia has a 25% chance of subsequently developing Wilms' tumour. Unawareness of this association can lead to a delayed diagnosis of Wilms' tumour. One such case in a 2 year old is reported. Wilms' tumour, one of the common childhood malignancies, is associated with other congenital anomalies in about 15% of cases. These include hemihypertrophy, genitourinary abnormalities, mental retardation, aniridia etc. Sporadic non-familial aniridia was noted in only 1.1% of 547 children with Wilms' tumours evaluated by the National Wilms' Tumour study group. Unawareness on the part of a clinician about these associated anomalies can lead to an avoidable delay in diagnosing Wilms' tumour. One such case in a two year old girl is being reported.


Investigation of the causes facilitating formation of microflora in atomic reactor pool water. In the course of investigations realized by us earlier it was found that there was no difference between radioresistance of microbes taken from various water sources. As a matter of fact quality of the microflora clearly reflects a unique phenomenon called selection which causes disappearance of all radiosensitive and survival of radioresistant types of microbes. There is indeed an increased number of radioresistant types of microbes with intensified activity of catalase and nuclease in pool water of atomic reactors.


Hemothorax from gestational trophoblastic disease: a case report. A 19-year-old woman who presented with right-sided hemothorax was diagnosed to have gestational trophoblastic disease with pleural metastasis. A CT scan of the chest revealed a pleural mass and serum Beta-subunit of HCG was high. After treatment the hormonal level became normal and the patient remains asymptomatic.


Human cytomegalovirus in immunoglobulin A nephropathy: detection by polymerase chain reaction. Human cytomegalovirus (HCMV) has been suspected to participate in the pathogenesis of IgA nephropathy (IgAN). However, with regard to the presence of HCMV in the renal tissue of IgAN, conflicting results have been reported using a variety of different techniques. Renal biopsies of 29 patients with IgAN, of 7 with focal segmental glomerulosclerosis (FSGS) and of 11 normal kidneys were analyzed for the presence of HCMV-DNA using the polymerase chain reaction. HCMV-DNA was detected by hybridization with digoxigenin-labelled probes in 14 of 19 analyzed frozen renal biopsies from patients with IgAN. However, only 1 of 17 renal biopsies of IgAN embedded in paraffin was positive for HCMV-DNA. Furthermore, HCMV-DNA was detected in 4 of 18 frozen normal kidneys but in none of the tissues from patients with FSGS. The present results provide further evidence of an association between the presence of HCMV in renal tissue and IgAN.


Peripheral adrenoceptors in hypotension of hemodialyzed uremic patients. Hypotension is a common problem in patients on hemodialysis. To further investigate this problem, the number of platelet alpha 2-adrenoceptors and the activity of lymphomonocyte beta 2-adrenoceptors were measured in 10 hemodialyzed patients with normal blood pressure and in 10 sex- and age-matched persistently hypotensive hemodialyzed patients. Density of alpha 2-adrenoceptors was assessed by the specific binding of 3H-yohimbine to intact platelets, while the function of beta 2-adrenoceptors was estimated by the production of cAMP after the exposure of lymphomonocytes to isoprenaline. The maximal number of alpha 2-adrenoceptors was increased in the hypotensive compared to the normotensive group (262.13 vs. 77.21 fmol/mg protein; p < 0.01). Plasma norepinephrine was higher in the hypotensive than in the normotensive uremic patients (640 +/- 195 vs. 344 +/- 156 pg/ml; p < 0.01). Plasma epinephrine did not differ in the two groups (90 +/- 30 vs. 94 +/- 24 pg/ml). The amount of cAMP, produced by stimulation of lymphomonocytes, was lower in the hypotensive than that in the normotensive uremic patients (7.7 +/- 2.4 vs. 15.6 +/- 5.4 pmol/10(6) cells; p < 0.002). The increased number of alpha 2-adrenoceptors together with a high level of norepinephrine and reduced activity of adenylate cyclase (coupled with beta 2-adrenoceptors) support the hypothesis that hypotension in the hemodialyzed uremic patients may be related to a defect in adrenoceptor coupling mechanisms.


Correlation between reduction of polymorphonuclear leucocytes in glomeruli injected with a newly developed monoclonal antineutrophil antibody and proteinuria in Masugi nephritis. The effects of the reduction of neutrophils in glomeruli on the improvement of proteinuria and glomerular injuries were determined in the first (heterologous) phase of Masugi (nephrotoxic) nephritis. Male (6-week-old) WKA/Hkm rats were initially injected with 2.0 ml of a newly developed monoclonal antineutrophil antibody and then injected with 1.0 ml of nephrotoxins. This monoclonal anti-neutrophil antibody was found to have selectively reduced the number of neutrophils in the glomerular capillary lumen in cases of Masugi nephritis by light microscopy. Malondialdehyde (MDA) levels or superoxide dismutase (SOD) activities in renal tissues of such rats were also examined. However, there were no significant differences in the levels of proteinuria and the number of glomerular cells containing resident cells and infiltrated mononuclear cells in the first phase of Masugi nephritis with or without pretreatment with antineutrophil antibody. No significant differences were observed in the levels of MDA or SOD activities in renal tissues of Masugi nephritis with or without pretreatment with such an antibody either. It appeared that infiltration of neutrophils in the glomeruli might not be related to proteinuria and glomerular injuries in the first phase of Masugi nephritis. It was postulated that the massive proteinuria in the first phase of Masugi nephritis might be correlated with the activities of reactive oxygen species induced by the glomerular cells, i.e. glomerular resident cells and infiltrated mononuclear cells.


Colocalization of receptors for vasoactive peptides on astrocytes of cultured rat spinal cord and brain stem: electrophysiological effects of atrial and brain natriuretic peptide, neuropeptide Y and bradykinin. Electrophysiological studies have been made of the actions of the vasoactive peptides atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), neuropeptide Y (NPY) and bradykinin (BK) on astrocytes in explant cultures of rat brain stem and spinal cord. Addition of ANP, BNP and NPY to the bathing solution at concentrations of 10(-8) and 10(-7) M caused depolarizations of the majority of astrocytes studied, some cells were hyperpolarized and on a small number of cells the peptides had no action. BK (10(-8) M) only induced depolarizations of almost all astrocytes studied. Our electrophysiological data provide strong evidence for the presence of receptors for these vasoactive peptides on astrocytes. When ANP, BNP, NPY and BK were tested on the same cell, all peptides were effective, suggesting a coexistence of peptidergic receptors on astrocytes.


Coexistence of paired helical filaments and glial filaments in astrocytic processes within ghost tangles. Ultrastructural examination of ghost tangles in an autopsy case of long-term Alzheimer's disease revealed, in addition to degenerate neurites containing paired helical filaments (PHF), astrocytic processes which included PHF. This finding suggests either that astrocytes in ghost tangles possess the capacity to produce PHF or that PHF are incorporated into astrocytes by endocytosis.


Fast sodium channel dependency of the somatodendritic release of dopamine in the rat's brain. Somatodendritic release of dopamine (DA) was studied by microdialysis. The basal release of endogenous extracellular DA in the rat's substantia nigra was 1.0 +/- 0.14 fmol/min (mean +/- S.E.M.; n = 6). Compared with the basal extracellular level of DA found in the striatum (12.9 +/- 0.94; n = 12), the former value was about 13 times smaller than the latter value. The addition to the Ringer solution of the fast sodium channel inhibitor, tetrodotoxin (TTX), at a concentration of 1 microM, produced the total disappearance of DA in the substantia nigra. When TTX was removed from the Ringer solution, the DA extracellular level came back slowly to the control value. This result suggests that the somatodendritic basal endogenous DA release measured by microdialysis is dependent on the sodium channel conductance.


Modulation by GABAB receptors of spontaneous synchronous activities induced by 4-aminopyridine in the rat hippocampus. Field potential recordings were made in the CA3 and/or CA1 subfields of rat hippocampal slices maintained in vitro during perfusion with medium containing the convulsant drug 4-aminopyridine (4AP, 50 microM). In this experimental condition, spontaneous interictal epileptiform discharges caused by the activation of excitatory amino acid receptors and synchronous, GABA-mediated potentials occurred regularly in both areas. Interictal discharges and GABA-mediated potentials were reduced and eventually abolished in both subfields by bath application of baclofen (0.5-100 microM), which is a selective agonist for the GABAB receptor. However, interictal epileptiform events disappeared in the presence of baclofen concentrations (i.e., 4.75 +/- 0.7 microM; IC50 = 3.4 microM; n = 9) that were lower than those required for abolishing the GABA-mediated potential (i.e., 96.1 +/- 19.4 microM; IC50 = 9.8 microM; n = 12). The effects of baclofen were antagonized by the GABAB receptor antagonists saclofen (1 mM; n = 3 slices) or CGP 35348 (0.2-1 mM; IC50 = 240 microM; n = 12 slices). Our findings indicate that activation of GABAB receptors by baclofen inhibits both types of 4AP-induced activities in the rat hippocampus although epileptiform discharges appear to be more sensitive than GABA-mediated potentials to this pharmacological procedure. Although our data do not indicate whether the action of baclofen is pre- or postsynaptic, they demonstrate that this mechanism is sensitive to available GABAB receptor antagonists.


Astrocytic localization of the messenger RNA encoding the isoenzyme B of inositol (1,4,5) 3-kinase in the human brain. The calcium-mobilizing second messenger inositol 1,4,5-trisphosphate (InsP3) is phosphorylated to inositol 1,3,4,5-tetrakisphosphate, another putative second messenger, through the activity of the enzyme InsP3 3-kinase. The cDNAs encoding two such isozymes have been recently isolated from a human hippocampal cDNAs library. We have previously reported the neuronal localization of the A form (Mailleux et al., Neurosci. Lett., 137 (1992) 69-71) and we here demonstrate the presence of the messenger RNA for the B form in human astrocytes.


Comparison of binding at strychnine-sensitive (inhibitory glycine receptor) and strychnine-insensitive (N-methyl-D-aspartate receptor) glycine binding sites. We compared, for a number of ligands to the two receptors, the displacement of [3H]strychnine binding to the glycine-gated chloride channel of spinal cord and brainstem synaptic membranes to the displacement of [3H]glycine binding to the NMDA receptor complex of hippocampal and cortex synaptic membranes. Glycine and beta-alanine are recognized by both receptors. In the NMDA receptor glycine antagonists, the kynurenic acids, most of the quinoxalinediones, and the (R)-enantiomer of HA-966 had little affinity at the strychnine-sensitive site. Surprisingly, the quinoxalinedione widely used as an AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) receptor antagonist, NBQX (2,3-dihydro-6-nitro-sulfamoylbenzo[f]quinoxaline-2,3-dione) displaced [3H]strychnine binding (IC50 = 11 microM) and to a lesser extent [3H]glycine binding (IC50 = 119 microM). Of the compounds tested, only strychnine, brucine, taurine and (S)-HA-966 were more potent displacers of [3H]strychnine than of glycine binding. Generally, the two glycine recognition sites appear to have remarkably different structural requirements.


Effects of Ca2+ antagonists and antiepileptics on tetrodotoxin-sensitive Ca(2+)-conducting channels in isolated rat hippocampal CA1 neurons. All Ca2+ antagonists blocked tetrodotoxin-sensitive Ca2+ current (TTX-ICa) more potently than Na+ current (INa). Phenytoin and MK-801, at concentrations which had no effect on INa, could block TTX-ICa concentration-dependently. Valproic acid and phenobarbital had no effect on both TTX-ICa and INa. In particular, flunarizine and phenytoin have more potent inhibitory effects on TTX-ICa than other test drugs. These results suggest that the abnormal excess-excitation of TTX-sensitive Ca(2+)-conducting channels may be one of the trigger factors generating epilepsy.


Enkephalin induces Ca2+ mobilization in single cells of bradykinin-sensitized differentiated neuroblastoma hybridoma (NG108-15) cells. A study of the intracellular Ca2+ ([Ca2+]i) response of differentiated neuroblastoma x glioma hybrid cells (NG108-15 cell) to enkephalin (EK) was carried out by fura-2 video-imaging. EK alone did not influence [Ca2+]i in single cells. The opioid did, however, induce a marked [Ca2+]i rise, when the cells were incubated with bradykinin (BK) prior to the EK treatment. Such BK-assisted stimulation of the differentiated hybridoma cells by EK was completely abolished by pertussis toxin treatment. These results suggest that in single NG108-15 cells, EK induces Ca2+ mobilization which is assisted by cross-talk between the EK and BK receptor systems via a pertussis toxin-sensitive G protein.


Influence of fixation on human papillomavirus DNA detection in frozen and embedded paraffin lesions by in situ hybridization with biotinylated probes. Series of frozen or paraffin-embedded tissues from various body sites, taken from non-immunosuppressed or immunosuppressed patients with persistent papilloma lesions were examined for the presence of group specific antigen from human papillomavirus (HPV) by indirect immunofluorescence or HPV DNA by in situ hybridization with biotinylated probes. We have shown that it is possible to detect HPV DNA after fixation of tissues in neutral formalin, Bouin's or Baker's solution. However, the sensitivity was reduced as compared to frozen tissues. The HPV DNA was detected in nuclei of heavily infected epithelial cells such as plantar or hand warts or in dispersed cells containing high copy numbers of HPV DNA from lesions such as squamous cell carcinomas or keratoacanthomas. In premalignant or malignant lesions of both immunosuppressed or non-immunosuppressed patients, HPV DNA was rarely detected after fixation. HPV types commonly described for skin and genital samples were identified in non-immunosuppressed patients, whereas in transplant recipients oncogenic HPV type 16 was identified in benign warts as well as in premalignant or malignant lesions. Positive reactions with several HPV types were more frequent in lesions from grafted patients than from the normal population. Virus antigen was detectable more frequently in frozen sections than in fixed tissues. Our findings indicate that in situ hybridization is an appropriate and rapid technique to study the presence of HPV infection. However, numerous controls are needed to avoid misinterpretations.


Signet-ring cell carcinoma of the stomach. A quantitative analysis on its submucosal invasion using a computer image analyzer. As has been indicated, signet-ring cell carcinoma often shows a three-layered pattern of mucus-containing cells in the mucosa, reproducing an organized differentiation in the normal mucosa. In 110 lesions of early stage signet-ring cell carcinoma of the stomach, the proportion between the area with the three-layered pattern and the non-layered area without such layered pattern was measured using a computer image analyzer. As the result, a positive correlation could be demonstrated between the extent of the non-layered area and the tumor size expressed in the largest diameter (r = 0.2777, P < 0.01). The incidence of submucosal invasion significantly increased when the non-layered area exceeded 50% of the total area of signet-ring cell carcinoma. The sites of submucosal invasion were almost always limited to within the non-layered area bearing another positive correlation between the area of submucosal invasion and that of non-layered intramucosal pattern (r = 0.4427, P < 0.01). Thus, it is indicated that the incidence of submucosal invasion in early signet-ring cell carcinoma was significantly related to a dissolution of the three-layered pattern of carcinoma within the mucosa. In addition, the presence or absence of fibrous and lymphoid stromal reactions to the carcinoma was evaluated in relation to the incidence of submucosal invasion.


Transformation of human colostrum lymphocytes with Epstein-Barr virus. Lymphocytes from human colostrum were transformed with EB virus to obtain immunoglobulin secreting cells. Equal proportions of IgG, IgA and IgM were detected in wells containing transformed colostrum cells. Pretreatment of colostrum cells with BCGF prior to EB virus infection reduced the transformation slightly. We discussed the possibility of establishing cell lines that produced human monoclonal antibody using colostrum cells.


Anti-HCV antibodies and hepatocellular carcinoma. Relationship in a medium-risk population. By means of an accurate immunoenzymatic assay, the prevalence was studied of antibodies to hepatitis C virus (HCV) in three different populations: 74 patients affected with hepatocellular carcinoma (HCC) on preexisting cirrhosis, 82 patients with liver cirrhosis but with no apparent neoplasm, and 70 control subjects, hospitalized for various conditions, of internal medicine or geriatric interest. 70.2% of HCC patients exhibited anti-HBC antibodies, versus 47.5% of cirrhotic subjects with no tumor and 7.1% of controls. Such results suggest the possible role of HCV in the etiopathogenesis of HCC, and its possible synergy with other agents-e.g., hepatitis B virus, alcohol--in causing chronically injured hepatocytes to become neoplastic.


[Diagnostic difficulties in a case of intracardiac tumor]. 64 years old female operated on for right auricular tumor, which manifested as right ventricular heart failure is presented. Although full preoperative diagnostics was performed the tumor appeared to be metastatic one with the primary focus situated in the liver (histologically ca hepatocellular). Necessity of cavography in case of right auricular tumor is stressed in the paper.


Reproductive failure associated with porcine parvovirus in an enzootically infected pig herd. Two outbreaks of porcine parvovirus occurred in a unit of 380 sows, with a subsequent decrease in the size of gilts' litters. Of the 203 gilts and 64 primiparous sows which were seronegative at the time of insemination, 134 gilts and 55 primiparous sows seroconverted during pregnancy. Of the second parity sows nine of 271 were still seronegative at the time of insemination but all nine seroconverted during their third gestation. The gilts that seroconverted during their first pregnancy produced on average 0.9 fewer live piglets than the gilts that did not seroconvert. Second litter sows which seroconverted produced 0.3 fewer piglets than those which did not seroconvert. There were no significant differences between sows which seroconverted or did not seroconvert in the numbers of returns to service, or in the percentages of piglets which were born dead or died before weaning.


[The effect of suloctidil and acetylsalicylic acid on eicosanoid synthesis in human platelets]. The study aimed at comparing an effect of suloctidil and acetylsalicylic acid on malonyldialdehyde levels resulting from the arachidonic acid metabolism in blood platelets. It was shown that inhibitory effect of suloctidil is more potent than that of acetylsalicylic acid. Therefore the first is more inhibitor of an enzymatic metabolism of arachidonate in blood platelets than the latter.


[The dynamics of the cerebral circulation during the surgical treatment of paragangliomas of the neck]. Clinical and rheoencephalographic studies ot 17 cases of paraganglioma of the neck revealed insufficient collateral blood circulation after compression of the common carotid artery on the affected side. In such cases, surgery carries a risk of brain ischemia. Improvement of collateral blood circulation was achieved by rheoencephalographically controlled compression of the common carotid artery performed daily with increasing duration. A patient was considered eligible for surgery if no signs of brain hemisphere ischemia were apparent following a 40% or less decrease in rheoencephalographic anacrotism amplitude from baseline. Common or internal carotid arteries were resected in 6 out of 17 cases of neck paraganglioma dissection. Adequate preoperative preparation prevented cerebral ischemia development in the postoperative period.


[Morphological diagnosis and prognosis in early ductal breast cancer]. One hundred and ten cases of precancer and early ductal breast cancer were analysed. Patterns of non-invasive ductal cancer were identified. Multicentric growth was observed in 55.8% of patients with intraductal cancer and invasive ductal cancer with predominant intraductal component. Early ductal cancer proved to have favorable prognosis, with only 2.2% of patients with disseminated tumor and 4.4% of those with recurrence identified during the mean follow-up period of 53.5 months.


[The possibilities of using the free polyamines of the peripheral blood as biochemical tumor markers in nephroblastoma in children]. Thin-layer chromatography was used to measure free polyamine level in blood plasma and peripheral blood cells in 38 pediatric nephroblastoma patients and 30 apparently healthy children. Thirty-six nephroblastoma patients were examined in the course of treatment of whom 25-during 1-36 months following basic treatment course as well. A significant increase in the initial free polyamine level in blood cells (92.4% of cases) and plasma (63%) of nephroblastoma patients was observed. A correlation was established between changes in free polyamine level, on the one hand, and response to treatment and subsequent course of disease, on the other. At 8-10 days following radical surgery, a decrease in polyamine level was observed in 58% of operated cases and an increase-in 41.8%. The latter proved prognostically unfavorable. Recurrence and metastases development was associated with a rise in polyamine concentration in plasma and, particularly, in blood cells which in some cases preceded the appearance of other signs of tumor progression. The polyamine test can be recommended for monitoring pediatric nephroblastoma patients.


[New aspects of adoptive immunochemotherapy in disseminated forms of cancer]. A method of autolymphochemotherapy was developed based on administration of a "therapeutic" course of chemotherapy following cytotoxic treatment of a large amount (1.5-2 l) of central lymph obtained by external drainage of the thoracic duct. Immediate results of treatment of 15 patients with advanced breast, lung (non-small-cell) and ovarian cancer with various degree of chemoresistance are presented. Marked response was observed in 14 cases. It seems justified to use the method for the treatment of locally advanced cancer.


[The morphological characteristics of primary breast cancers with different growth rates]. Morphologic peculiarities of primary breast cancer versus growth rate were studied in 46 cases. Growth rate (actual doubling time) was established by mammography carried out at certain time intervals. Morphologic investigation included assessment of tumor histology, degree of invasion, growth pattern, presence of pathologic mitoses, degree of blood vessel invasion, degree of necrosis, presence of microcalcinates, type of desmoplastic reaction, peculiarities of cell infiltration, background and regional lymph node reaction, and grade of malignancy. None of the above parameters appeared to correlate with tumor growth rate. It is concluded that standard morphologic examination of tumor be supplemented with assessment of its proliferative activity.


[The combined chemotherapy with platinum of patients with inoperable non-small cell lung cancer]. Combination chemotherapy with cyclophosphamide, 5-fluorouracil, adriamycin and platinum was performed in 122 patients with stage III-IV squamous-cell cancer and adenocarcinoma of the lung. Control group included 84 cases of non-small-cell lung cancer treated with the same drugs except platinum. Ten patients with stage III disease received platinum-based combination chemotherapy and split-course radiotherapy in the total dose of 65-70 Gy. Complete or partial remission was obtained in 48.6 +/- 11.74% of stage III patients following three cycles of chemotherapy, with mean survival reaching 8.6 +/- 3.2 months. Survival in responders was 10.6 +/- 2.1 months. Chemoradiation treatment was associated with even higher survival (14.6 +/- 2.16 months). It is concluded that platinum be used in combination chemotherapy and chemoradiation treatment of the above categories of patients.


[The effect of mefenamic acid on the immunity indices and hemostatic system in cancer patients and on the cathepsin D-like protease activity in the tissues in cancer of the large intestine]. The study of the effect of sodium mefenaminate on radiation resistance of rats yielded positive results. Clinical investigations showed mefenamic acid to decrease the activity of cathepsin D-like protease in colonic cancer tissue. The acid failed to affect the proteolytic activity of the normal mucosa. It revealed an immunomodulating activity and influenced the hemostatic system which usually manifested itself in amelioration of hypercoagulation.


[The degree of tumor vascularization as a prognostic factor of the efficacy of neoadjuvant polychemotherapy in breast cancer]. A procedure for obtaining microcirculation coefficient is described. It is based on the phenomenon of formation of a focus of radionuclide hyperfixation on scintigrams of tumor area due to embolization of its arterioles and precapillaries with radionuclide. The method was used in 26 breast cancer patients before neoadjuvant chemotherapy. A direct correlation between the microcirculation coefficient and degree of tumor-induced pathomorphosis of tumor cells was established. The coefficient values proved more or less predictive of tumor response to chemotherapy.


[The clinical assessment of the results of predicting individual sensitivity to adjuvant chemotherapy in lung tumors]. The value of subrenal capsule assay for predicting individual chemoresistance of tumor was evaluated using samples from 32 lung carcinomas. Treatment with drugs which had proved effective in the test was followed by a recurrence-free period of 15.5 +/- 3.7 months, as compared to 8.0 +/- 1.0 months for patients receiving ineffective drugs.


[Role of ACE in the gastric cytoprotection. Dopaminergic and peripheral alpha 2 adrenergic mechanisms]. In Wistar rats, four experiments were carried out, studying specific inhibitors for ACE (angiotensin-converting enzyme), such as captopril, enalapril, lisinopril, ramipril and cilazapril, in presence of 20% and 95% ethanol induced gastric lesions. The effect of lisinopril and angiotensin I on the same injury was also studied. Subsequently, drugs with known role in gastric mucosa cytoprotection, such as enprostil, paracetamol, ketotiphen, levamisole, diazepam, bromocriptine, dopamine and clonidine, before and after absolute ethanol gastric injury were also studied. Finally, to examine the potential role of ACE in gastric cytoprotection, pretreatment with enalapril, followed by cytoprotective drugs, was carried out. We conclude that all ACE blockers aggravate the gastric lesion induced by 20% and 95% ethanol and are similar to the findings with angiotensin I. The gastric ACE probably plays an important role in the gastric mucosa defense. On the other hand, gastric ACE, as a physiologic regulator of microcirculation, acts by a double mechanism. a) converting the vasopressor amine, angiotensin I in angiotensin II. b) activating vasoactive peripheric receptors such as DA 2 dopaminergic and alfa 2 adrenoreceptors.


[Re-examination of the role of mast cells, mediators of inflammation and heparin]. Mast cells contain proteoglycans, e.g. heparin, which manifest potent anti-inflammatory features. Mast cell degranulation could eventually result in self limiting inflammation with shortened course due to heparin release. The recent availability of low molecular weight heparins will allow for clinical studies in allergic/inflammatory syndromes, e.g. asthma.


[Comparative study of the therapeutic effect of the glycopeptide antibiotics eremomycin, vancomycin and ristomycin in a model of antibiotic-associated colitis in golden hamsters]. Experiments with a model of intraperitoneal or intragastric lincomycin-induced fatal colitis indicated that eremomycin, vancomycin and ristomycin administered orally in daily doses of 100, 100 and 200 mg/kg, respectively, for 5 days protected the animals from development of antibiotic-associated colitis (AAC), which was evident from prolongation of their life-span to 10-23 days against 3-9 days in the controls. Eremomycin administered intraperitoneally according to an analogous scheme protected the animals from development of AAC, prevented 45 per cent of the animals from death and prolonged the life-span of the other animals to 15-28 days against 3-9 days in the controls. Vancomycin administered intraperitoneally was somewhat more efficient. Still, unlike eremomycin it had a local irritating effect. The protective effect of ristomycin administered intraperitoneally was much lower than that of vancomycin and eremomycin.


[Antiviral properties of various pharmacologic groups of drugs]. Currently used cardiovascular drugs such as nicotinamide, strophanthin, corglycon, curantyl, cavinton, papaverin hydrochloride, nicotinic acid, xantinole nicotinate, isoptin, parmidin and halidor were studied by the program of antiviral drug screening. The majority of them (9 out of 11) were shown to have antiviral activity which was rather individual by its specificity and level. Laboratory strains of 9 viruses inducing the most common infections in man and animals, i.e. Herpes simplex, poxvaccine, influenza, vesicular stomatitis, respiratory syncytial infection, VEE, ECHO. Lassa fever and rotavirus infection were tested. The characteristic feature of the drugs was their high specific activity against the DNA-containing viruses and rotavirus. The three drugs papaverin hydrochloride, strophanthin and corglycon proved to be the most promising. Their antiviral activity was confirmed on a model of herpes infection in mice. The paper discusses whether the phenomenon discovered in the official drugs is important in the therapy of somatic patients.


Synthesis and immunological activity of dialkyloamine- and N-piperidinederivatives of dithiocarbamic acid. A series of aminocarbamic acid derivatives, containing fragments of substituted hydrazine and dithiocarbamic acid, was synthesized. The immunopharmacologic studies showed these that derivatives exerted immunotropic, mainly suppressive effects (PFC, circulating immunoglobulins, E-RFC, A-RFC). Some of these compounds, however, caused significant increase of weight of the popliteal lymph nodes. The immunotropic activity of the newly synthesized preparations is comparable with the action of sodium diethyldithiocarbamate (imuthiol).


The cytokines effect on EBV-immortalized human B cells producing antisperm antibody. We have analyzed the antisperm antibody production of autoimmunized male subjects using Epstein-Barr virus (EBV) immortalization of B lymphocytes. We evaluated the influence of several in vitro culture variants applied prior to EBV infection on the frequence of antibody-producing cells and affinity of secreted antibodies. The following variants were applied: a) polyclonal antigenic stimulation of lymphocytes with PWM, b) PWM (pokeweed mitogen) + IL-2 + interferon gamma and c) PWM + IL-2 + interferon gamma + sperm antigenic extract. The variants where the cytokines were added did not increase the frequency of EBV-infected antibody-producing cells as comparing to EBV infection previously amplified by the use of polyclonal activator. Furthermore the cytokine activation either in combination with mitogen or in vitro secondary antigenic sensitization (prior to EBV transformation) did not seem to have beneficial effect on affinity of antibodies produced by EBV-infected cells in comparison to straight EBV infection. On the other hand, the attempt to promote an immunoglobulin secretion (IgM) by previously obtained human-human antisperm hybridomas by adding of IL-2 was quite successful.


[Structural studies of cDNA and the gene for the beta-subunit of cGMP phosphodiesterase from human retina]. cDNA clones encoding the beta-subunit of the photoreceptor cGMP phosphodiesterase-(PDE) were isolated from a human retinal library. The encoded polypeptide has 854 amino acid residues with calculated molecular mass of 98416 Da. Alignment of the deduced amino acid sequence with the previously analysed alpha-, beta- and alpha'-subunits of the bovine and mouse PDEs demonstrates highly significant similarities. We have also isolated, from a genomic library, two overlapping recombinant lambda phage clones containing 26 kb of the human PDE beta-subunit gene. The cloning of the human gene and the knowledge of its genomic organization will allow the rapid assessment of the role of this gene in the causation of human retinopathies.


Connexin expression and gap junction communication compartments in the developing mouse limb. Fundamental to the understanding of mouse limb morphogenesis and pattern formation is the need to elucidate the spatial and temporal distribution of gap junction proteins (connexins, Cx) and cell-cell communication compartments. To this end, we used immunofluorescence and confocal microscopy together with 3-dimensional reconstruction software to map the distribution of Cx43 and Cx32 in 11-14.5 days postcoitum (dpc) mouse limbs. Cx43 was strictly localized to the apical ectodermal ridge (AER) and nonridge ectoderm throughout all stages of mouse limb development studied. Cx32, on the other hand, was abundant in the mesenchyme with only low levels of expression in the 11-13.5 dpc ectoderm. However, at 14-14.5 dpc there was a clear increase in Cx32 expression in the ectoderm. Double labeling for connexins and confocal microscopy revealed Cx43 and Cx32 in the same optical section of the basal cells of the ectoderm but in separate plaques. Lucifer yellow dye injections showed that the cells of the AER were in direct communication with the nonridge ectoderm but dye was never observed to spread to the mesenchyme. Cells of the mesenchyme were coupled to each other but to a much lesser extent than cells of the ectoderm. Finally, although there was an increase in Cx32 expression in the ectoderm at 14-14.5 dpc, this was not correlated with any detectable change in communication compartments. Thus, the lack of dye transfer between the ectoderm and underlying mesenchyme from the peak of AER height through its decline suggests that bulk transfer of morphogens between these two layers is not necessary for mouse limb development.


Somatic mutation of the APC gene in gastric cancer: frequent mutations in very well differentiated adenocarcinoma and signet-ring cell carcinoma. We searched for somatic mutations of the adenomatous polyposis coli (APC) gene in DNA samples isolated from 57 sporadic gastric cancers, by means of a ribonuclease (RNase) protection analysis coupled with DNA amplification by the polymerase chain reaction (PCR). Examining 30% of the APC coding region, including a region where somatic mutations in colorectal tumors are known to be clustered, we detected somatic mutations in 12 tumors; seven in 17 very well differentiated adenocarcinomas, two in 19 well or moderately differentiated adenocarcinomas, and three in ten signet-ring cell carcinomas. So far, no somatic mutations have been identified in 11 poorly differentiated adenocarcinomas. Eight of the 17 somatic mutations found in 12 tumors caused truncation of the gene product due to a nonsense mutation and a 1-, 2- or 5-bp deletion; nine others were point mutations that altered amino acids. Our results suggest that inactivation of APC plays a role in development of some gastric cancers, particularly very well differentiated adenocarcinomas and signet-ring cell carcinomas.


[Equivalent dose of cosmic rays at representative points in models of the human body]. Equivalent doses of cosmic ray protons were studied in a wide range of spectra realized in solar cosmic rays and Earth radiation belts, for three standardized models of the human body. To solve this task, a problem was created for calculating the function of shielding of representative points within an anthropomorphic model. Equivalent dose values in various models were compared. It was shown that representative points of central nervous and hemopoietic systems in a spherical model of the human body did not yield coordinated values of equivalent dose as compared to stricter models. It is recommended to clarify the standard position of the mentioned points.


Localization of the synovial sarcoma t(X;18)(p11.2;q11.2) breakpoint by fluorescence in situ hybridization. A high proportion of synovial sarcomas contain a chromosome translocation t(X;18)(p11.2;q11.2). We have previously used somatic cell hybrids derived from an established cell line, SS255, to map the X chromosome breakpoint to the interval flanked by the markers DXS14 and DXS146. In this study we have examined these hybrids with thirteen additional markers located at Xp11.3-Xcen, by Southern hybridization. Based on these results we have delimited the breakpoint as follows Xpter-DXS228-(UBE1-OATL1-TIMP-DXS226 )-(DXS255-TFE3-ELK1-DXS146)-OATL2- X;18-(DXS14-DXS422-DXS423-DXS674-DXS679)-+ ++Xcen. Confirmation of the breakpoint location has been obtained by analysis of two synovial sarcoma cell lines, SS255 and HA2243, using fluorescence in situ hybridization. A 350kb YAC probe spanning the DXS423 locus hybridized only to the derivative X chromosome, showing that it maps proximal to the breakpoint. Two YAC probes of 300kb and 450kb, containing the OATL2 locus, hybridized to both derivative chromosomes, indicating that these YACs span the translocation breakpoint. Similar results were obtained with both cell lines. The identification of YACs that span the t(X;18) breakpoint now facilitates a strategy for cloning candidate genes from this precisely defined region.


Expression of a cytomegalovirus IE-1-factor VIII cDNA hybrid gene in transgenic mice. A construct containing the 5' end of the human cytomegalovirus major immediate early gene fused to the human coagulation factor VIII cDNA was used to produce transgenic mice. Two out of five transgenic lines transcribed the construct. The expression was consistently seen in a limited number of tissues and was highest in muscle tissues. This is in contrast to the almost ubiquitous activity demonstrated in earlier studies with the IE-1 enhancer/promoter. Human factor VIII protein was detected immunochemically in muscle tissues at levels several times higher than in human plasma.


Inherited haemolytic anaemia created by insertional inactivation of the alpha-spectrin gene. In the process of generating transgenic mice, inserted foreign DNA can cause insertional inactivation of the flanking genetic locus and simultaneously provide a molecular tag for localizing and cloning the inactivated gene. We describe the case of an insertional mutation leading, in animals homozygous for the insertion, to severe anaemia that was lethal within a few days after birth. The haemolytic anaemia and microspherocytosis of the red cells strongly suggested membrane abnormalities of the erythrocytes. By in situ localization of the integration site, protein analysis of the red cell membranes, northern and Southern blot analyses, we were able to demonstrate that the integrated transgene had affected the alpha-spectrin gene locus.


Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Progress in plant molecular biology has depended heavily on the availability of effective vectors for plant cell transformation and heterologous expression. In this paper we describe the structures of a wide array of plasmids which have proved extremely effective in (a) plant transformation, (b) expression of heterologous genes and (c) assaying the activity of transposons in transgenic plants. Constructs that confer resistance to kanamycin, hygromycin, streptomycin, spectinomycin and phosphinotricin, or that confer beta-glucuronidase (GUS) gene expression are presented. Binary vector constructs that carry polylinkers of the pUC and Bluescript types are also described. Plasmids that permit the expression of any heterologous reading frame from either nopaline synthase (nos) or octopine synthase (ocs) promoters, as well as the cauliflower mosaic virus 35S promoter, using either the nopaline synthase or octopine synthase 3' polyadenylation sequences, are presented. These constructs permit a choice of orientation of the resulting transgene of interest, relative to the orientation of the selection marker gene. Most of the plasmids described here are publicly available.


[Na+/H+ exchangers, alpha-2-adrenergic receptors, sodium sensitivity and arterial hypertension]. Existing evidences indicate that a crossed regulation between alpha 2-adrenergic receptors and Na+/H+ exchanger(s) exists, that Na decreases the affinity of alpha 2-adrenergic receptors for agonists and antagonists, that intracellular Na+ and H+ ion concentrations regulate Na+/H+ exchanger activity, that intracellular pH controls the affinity of the alpha 2-adrenergic receptors for their agonists and antagonists. Alterations of alpha 2-adrenergic receptor densities and allosteric regulation by sodium have been demonstrated in sodium-dependent hypertension in rats. Increased Na+/H+ exchanger activity has been reported in genetic hypertension. Nevertheless, cosegregation experiments and human genetic polymorphism suggest that the exchanger could not be related to hypertension. We propose the following hypothesis: the increased Na+/H+ exchanger characteristic of hypertension could be secondary to the abnormalities of the alpha 2-adrenergic receptors found in hypertension, probably through the alteration of the sodium allosteric effect on these receptors.


[Nuclear receptor(s) regulating peroxisome genes]. Peroxisomes are becoming more and more attractive. This interest has grown up due to some key metabolic functions (i.e. very long chain fatty acid specific beta-oxidation, prostaglandins and polyamines catabolism, and first reactions in plasmalogen synthesis, as well as biliary salts) and to their strong response to peroxisome proliferators which can promote hepatocarcinogenesis in rodents. This mini-review comments recent breakthrough which is the discovery of peroxisome proliferator activated nuclear receptors (PPAR's) from the steroid receptor superfamily. Mouse PPAR and Xenopus PPAR's (alpha, beta and upsilon) activate the rat peroxisomal acyl CoA oxidase gene by recognition of specific responsive elements containing the 5'flanking "TGACCT" repeated sequence in the -560 upstream region. Such mechanism is involved in the control of peroxisomal beta-oxidation pathway.


[Anti-HBs-F(ab)'2 to the seroconversion of HBsAg carriers]. The exogenous anti-HBs-F(ab)'2 was given to the HBsAg carriers for the study of its effect of seroconversion in vivo. 30 HBsAg carriers, with a duration of 2-15 years (HBeAg positive 18 and anti-HBe positive 12) were studied after intramuscular infection of anti-HBs-F(ab)'2. In the HBeAg group, the cut off value of HBeAg decreased significantly (P < 0.01) in 18 carriers, of which 6 seroconversed to anti-HBe, all the HBe-Ag carriers had a high S/N ratio of anti-HBs-anti-Id (RIA) but after 12-week treatment the ratio decreased significantly (P < 0.01). Six of 18 HBV DNA positive carriers conversed to negative (33%) and 8 of 13 DNA-P positive conversed to negative (61%). 12 anti-HBe positive carriers showed no seroconversion.


[Hepatitis B virus infection in monocytes and their functional changes in patients with chronic hepatitis B]. We detected the HBV DNA in peripheral blood monocytes of 70 patients with chronic hepatitis B and 20 healthy controls by polymerase chain reaction (PCR) and Southern transfer hybridization technique, the HLA-DR expression on monocytic membrane and tumor necrosis factor-alpha (TNF alpha) content and interleukin 1 (IL 1) activity in their monocytic culture supernatant were also measured respectively by indirect immunofluorescence technique, ELISA, and thymocytic multiplication assay. HBV DNA sequences were detected in monocytes of 42(60%) of the 70 patients. The percentage of monocyte-expressing HLADR and the TNF alpha content in monocyte culture supernatant were significantly elevated in patients with HBV DNA-positive monocytes compared with those patients with HBV-DNA-negative monocytes. The percentage of monocyte-expressing HLA-DA and the level of TNF alpha produced by monocytes correlated markedly with the intensity of HBV DNA-positive signal in monocytes. These data suggested that HBV infection in monocytes might cause a series of functional changes in these cells.


[Experimental infection with hepatitis E virus in rhesus monkeys]. An experimental model of hepatitis E was established in Rhesus Monkeys. Four animals were inoculated with stool suspensions that obtained from patients with hepatitis E. Two animals injected with PBS were taken as negative controls. After 4-6 weeks the ALT of the four animals infected HEV raised 3-4 times and the live biopsy showed acidophilic degeneration of hepatocytes and coagulative liver cell necrosis. Acidophilic bodies were frequently observed. Foci of lytic single-cell necrosis were also observed. Degenerative and microinflammatory changes in liver parenchyma in zones 1-2 of liver lobules were accompanied by inconspicuous infiltrations in portal tracts. 27-34 nm virus-like particles were recovered from the stools of infection animals by IEM.


[Dynamic changes of striatal monoamines with in vivo microdialysis in ischemic rats]. Using intracerebral microdialysis, high performance liquid chromatography and electrochemical detection (HPLC-ECD), we determined the contents of extracellular dopamine (DA), serotonin (5-HT) and their metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid(HVA) and 5-hydroxy-indole acetic acid (5-HIAA) in the striatum of rats after the occlusion of middle cerebral artery (MCAO). The dialysis tube was implanted stereotaxically in the lateral 2/3 of the striatum. No significant effects of stressors were noted on the concentration of neurotransmitter mentioned above. A step rise of extracellular DA and 5-HT accompanied by significant reduction of DOPAC, HVA and 5-HIAA was found 10 minutes after MCAO. From then on, the enhanced concentrations of DA and 5-HT decreased gradually. At 3 hours after MCAO, however, DA and 5-HT were still high with respect to the basal levels. Their metabolites were kept at low levels at 3 hours of MCAO. In conclusion, the massive leakage of DA and its over production of oxygen free radicals are important for the development of ischemic cell damage in the striatum.


[Interleukin 6 production and interleukin 6 mRNA gene expression by mouse glomerular mesangial cells]. To investigate the relationship between Interleukin 6(IL 6) and glomerular mesangial cells (MC), the MC of BALB/c mouse were cultured and verified subsequently by anti-keratin and antidesmin antibodies. The cultured MC were divided into two parts those with Mac-1 marker and those without by flow cytometry. The IL 6 activity of cultured supernatants of MC was measured using the IL 6-dependent cell line subclone (MH 60). The results show that the mouse MC can produce IL6 and the level of IL 6 produced by MC with Mac-1 marker was higher than that without Mac-1. The RNA from the mouse glomerular mesangial cells was hybridized in dot blots with a-32pdCTP labeled IL6 cDNA probe, the dot blot autoradiograph showed the expression of IL6 mRNA gene in mouse mesangial cells.


[Effects of prostaglandin E2 on growth and function of osteoblasts in human cell culture in vitro]. The effects of prostaglandin E 2 (PGE2) on the differentiation and proliferation of osteoblasts (human fetal bone-cells) cultured in serum-free medium were investigated by assays of alkaline phosphatase (ALP) activity, intracellular cyclic AMP level and collagen synthesis in the cells. The results suggested that PGE2 in physiologic concentration stimulated the differentiation of osteoblasts in vitro, and might be involved in bone formation in vivo.


[Frequencies of 6 known mutations in phenylalanine hydroxylase gene and their application in prenatal gene diagnosis]. The known mutant alleles of human phenylalanine hydroxylase gene were analyzed in 25 phenylketonuria (PKU) families from North China by polymerase chain reaction combined with allele-specific oligonucleotide dot hybridization techniques. The results showed that the six mutation accounted for 62% of all PKU genes. The three most frequent mutations were Arg243-->Gln in exon 7, Arg413-->Pro in exon 12 and Tyr204-->Cys in exon 6. They accounted for 24%, 18% and 16% respectively of all PKU alleles. The frequencies of Arg111-->Ter mutation in exon 3, Tyr356-->Ter mutation in exon 11 and Trp326-->Ter mutation in exon 10 were somewhat lower in North China. They accounted for 4% of all PKU alleles. Among these 25 PKU families, prenatal gene diagnosis was possible in 10 families. In 11 families, 50% exclusive diagnosis was possible. Prenatal gene diagnosis was performed and confirmed in 2 families.