Cerebellar granular layer degeneration in small cell lung cancer: paraneoplastic cerebellopathy or artifact? The aim of our study was to ascertain whether granular cell degeneration represents uniquely an artifactual or a supravital event in patients with oat cell carcinoma. The material includes 52 cases of small cell lung cancer (SCLC). Formalin fixed and paraffin embedded representative cerebellar slides were stained routinely (HE, Klüver-Barrera), and some of them served as material for immunohistochemical study. The following antibodies were used: anti-ferritin, anti-GFAP, anti-IgG and anti-C3 complement fraction. Finally 5 cases out of our material could be diagnosed as paraneoplastic cerebellar degeneration (PCD), on the basis of lack of metastases within the CNS and concomitant intensive loss of Purkinje and granule cells. Clinically the cerebellar syndrome was disclosed in 3 cases. In the granular layer prevalence of microglial cell reaction was noted. GFAP-labeled astroglia were not demonstrated in the same intensity. Antisera to the C3 complement fraction showed moderate staining of Purkinje cell cytoplasm and in some cases also of granule cells. IgG immunostaining was disclosed in Purkinje cell cytoplasm and in 4 cases also in granule cell nuclei. The immunopathological changes presently observed and glial cell proliferation could be evidence for a nonartifactual origin of PCD.


Incidence of feline immunodeficiency virus reactive antibodies in free-ranging lions of the Kruger National Park and the Etosha National Park in southern Africa detected by recombinant FIV p24 antigen. Lion sera from the Kruger National Park (KNP) dating back to 1977 and from the Etosha National Park (ENP), obtained from 1989 to 1991, have been analysed by ELISA and Western blot analyses using a genetically engineered antigen representing the p24 structural protein of feline immunodeficiency virus (FIV). It was concluded that some 83% of 98 KNP lion sera reacted with the p24 antigen, while none of 28 ENP lion sera reacted. A few other KNP felids (cheetahs and genets) gave samples that did not react with the FIV p24 antigen. For the KNP lions, apart from a lower prevalence in cubs (50%), no particular trends were demonstrated in terms of age, sex, date or origins of the samples. In Western blot and radio-immunoprecipitation analyses the lion sera reacted with the engineered p24 antigen, as well as with the p15 and p24 gag proteins and the p50 gag precursor protein from FIV, indicating that the agent is probably a lentivirus related to FIV. The ELISA with the engineered p24 antigen required less serum and appears to be more sensitive at detecting FIV-reactive antibodies than assays with available commercial kits.


Bradykinin receptors: pharmacological properties and biological roles. Kinins contribute to the acute inflammatory response and are implicated in the pathophysiology of inflammatory disease. The development of therapeutically viable agents that counteract the effects of kinins is, therefore, potentially very rewarding. Since kinin actions are generally mediated via an interaction with cell-surface receptors, one approach is the development of site-specific receptor antagonists. The emphasis in this review is to outline our current understanding of the properties of bradykinin receptors and the potential therapeutic applications for drugs acting at these sites. As a result of the recent introduction of potent bradykinin receptor antagonists and the cloning of bradykinin receptor genes, considerable advances in kinin research can now be confidently anticipated.


Isolation of Getah virus from mosquitos collected on Hainan Island, China, and results of a serosurvey. An isolate of Getah virus was obtained from Culex mosquitos collected in Mao'an Village, Baoting County, Hainan Province, China, in 1964. The virus (strain M-1) replicated in laboratory-bred Aedes aegypti and Cx. fatigans (= quinquefasciatus), and was transmitted by laboratory-bred Ae. albopictus to healthy newborn albino mice. Skeletal muscles of newborn albino mice experimentally infected with the virus showed degeneration, atrophy, necrosis, and inflammatory changes of muscle fibers. Antibody prevalence in humans and animals ranged from 10.3% by neutralization tests of samples from healthy people in 1979 to 26.4% by CF tests of samples from people with febrile illnesses in 1982. The high prevalence of antibody in pigs, horses, and goats (17.6% to 37.5%) indicated that infection with Getah or a closely related virus is relatively common in domestic animals.


Acute extradural hematoma associated with silastic dural substitute: case report. The author reports the occurrence of an acute extradural hematoma associated with a silastic dural substitute, originally implanted 13 years previously. A recurrent malignant meningioma also arose at the site of the previous surgery.


Dietary factors influencing protein utilization: a review. Utilization of dietary protein depends not only on the amino acid composition and the digestibility of the protein itself, but also upon the level of protein, feed intake, energy, fibre, and other constituents in the feed. On the other hand, the animal species, sex, age and health status may also play an important role. A deeper understanding of these factors helps to utilize feedstuffs as a protein source more efficiently.


[Experimental infection of pregnant gilts with Aujeszky's disease virus strain RC/79]. The RC/79 strain of the Aujeszky's disease virus was able to induce reproductive failure of pregnant gilts intranasally inoculated at different gestation periods. Four gilts 40-46 days pregnant (group A) and 6 gilts 70-73 days pregnant (group B) were instilled with 0.2 ml x 10(5) tissue culture infectious dose 50 (TCID50/0.2 ml) of the RC/79 strain into each nostril. Two gilts 70-73 days pregnant (group C) were used as non exposed controls. The three groups were kept in separated boxes and they were observed for clinical signs of infections and samples were collected for determination of viral shedding every day. Viral isolation was attempted in Vero cells (figure 1). From the 2nd to 7th day after inoculation, groups A and B showed fever anorexia, sneezing, coughing and depression; and viral isolation from nasal swabs was possible in 7 gilts at days 4 to 11, 9 gilts developed neutralizing antibodies. The virus caused fetal reabsorption in swine during the first period of pregnancy (group A), while infection during late pregnancy resulted in still birth or normal pigs and one mummification (group B). The entire a live litter was composed of no more than 8 suckling pigs in both groups. At necropsy virus from turbinates, ovary , placenta, spleen and lung could be isolated only from 3 gilts (group B, table 1). In 5 of 35 stillbirth and alive fetuses virus could be isolated from spleen (100%), lung (80%), liver (60%) and brain (40%) indicating that the virus has the ability to cross the placental barrier thus producing lesions in porcine fetuses and causing reproductive failure in sows (table 2). Tissue specimens from these 35 fetuses were fixed in 10% formalin, included in paraffin sectioned and stained with hematoxylin and eosin. In 13 fetuses microscopic lesions i.e. necrotic foci were found in lung (60%), liver (40%) and spleen (20%), these alterations were coincident with gross lesions in most of them. Inclusion bodies were absent. The gilts organs did not present gross lesions.


Malignant fibrous histiocytoma of the auricle: an immunohistochemical and electronmicroscopic study. Malignant fibrous histiocytoma (MFH) is a soft-tissue tumor of late adult life. This tumor is uncommon in the head and neck region. A case of MFH arising in the auricle is presented with immunohistochemical and electronmicroscopic findings. Histologically the tumor was characterized by numerous spindle cells with abundant eosinophilic cytoplasm and many small capillaries. They were positively stained with vimentin, alpha 1-antitrypsin, and alpha 1-antichymotrypsin. Electronmicroscopically, some tumor cells contained abundant lysosomes and others contained numerous filaments.


[The renin-angiotensin system and anterior pituitary function: endocrine interaction or paracrine modulation?]. The identification of elements of the renin-angiotensin system and ulterior demonstration of specific cellular angiotensin II (Ag II) receptors at hypothalamic-pituitary level has suggested the possibility that these peptide play a modulatory role on anterior pituitary secretion. Recent "in vivo" and "in vitro" studies provided confirmation of a stimulatory effect of the renin-angiotensin system on growth hormone (GH) and ACTH variable. Initial experiments indicate that there is no effect on TSH and gonadotrophins secretion. Treatment with angiotensin-converting enzyme inhibitors leads to a reduction of Ag II levels that results in slight decrease of PRL and GH concentrations. However, the mechanism by which Ag II is able to modulate anterior pituitary secretion still remains obscure. Both, indirect effects mediated through changes in neuropeptide secretion or a direct interaction on pituitary cells by a paracrine or endocrine mechanism emerge as possible hypothesis to explain this regulatory function.


Maximal .OH production is seen upon reoxygenation of viable anoxic cultured cardiomyocytes but not of compromised cells. Hydroxyl radicals (.OH) in isolated cultured cardiomyocytes upon reoxygenation (reoxy) were measured after graded anoxia (A) using high performance liquid chromatography (HPLC). Isolated myocytes were subjected to A for 15, 30, 60, 90 and 120 minutes and reoxy for 120 seconds. Supernatant was collected after reoxy, extracted with ether and injected into HPLC for measuring hydroxylation products of salicylic acid (2,5-DHBA) as an indicator of .OH formation. 2,5-DHBA was detected maximally after 15 minutes of A and 120 seconds of reoxygenation (34.2 +/- 3 pmol/mg protein), at which time 80% of cells had maintained their rod shape and 99% of cells excluded both trypan blue (TB) and horseradish peroxidase (HP). There was significantly less (P < 0.05) 2,5-DHBA in the group subjected to 15 minutes of A only without reoxy (7.95 +/- 1.2 pmol/mg protein). 2,5-DHBA decreased to 13.1 +/- 2 pmol/mg protein at 120 minutes of A/120 seconds reoxy. With increasing anoxic time, the number of rod-shaped cells decreased from 80% at 15 minutes to 30% at 120 minutes, while the number of TB/HP positive cells increased from 0% at 15 minutes to 100% at 120 minutes. The cell membrane blebs were nonexistent at 15 minutes, but at 120 minutes A intense bleb formation was observed. These data suggest that .OH is produced upon reoxygenation of anoxic cultured cardiomyocytes and their production is maximum when majority of myocytes are viable.


Ultrastructural effects of hydrogen peroxide on the sarcolemma of rat heart. Ultrastructural effects of hydrogen peroxide (H2O2) on the sarcolemma of the isolated rat heart were investigated with transmission electron microscopy combined with biochemical, enzyme histochemical, and freeze fracture techniques. Three hundred microM H2O2 were continuously administered to the Langendorff perfused isolated rat hearts. A significant amount of lipid peroxidation associated with depressed Na-K-ATPase activity was observed after 15 minutes of H2O2 perfusion (Group I), and consequently the cell membrane permeability was greatly increased. When 2.5 mM,N'-diphenyl-1,4-phenylenediamine (DPPD), a potent antioxidant, was added to the perfusate, the lipid peroxidation was totally inhibited (Group II). DPPD prevented an increase in the cell membrane permeability. However, Na-K-ATPase activity was not restored by DPPD. Decreased cytochemical staining of Na-K-ATPase was associated with an increase in cell membrane permeability. H2O2 appears to affect, not only lipids but also, intramembranous proteins embedded in the cell membrane. The combined effects of H2O2 on the membrane lipid and proteins result in the formation of membranous blebs.


[Detection of HPV-16-related DNA sequence in laryngeal squamous cell carcinoma]. The gene libraries of laryngeal squamous cell carcinoma (LSCC) and laryngeal papilloma (LP) were screened by hybridization analysis with alpha-32P-labelled HPV-16 DNA under low stringency (Tm = 40 degrees C). It showed that the relative sequences were detected in LSCCs (11/16, 68.8%), but not in LP. The HPV-16-related DNA harbored in negative hybridization lesions were further analysed by polymerase chain reaction to amplify E6/E7 gene of HPV-16-related DNA. The results showed that 1/5 LSCCs and 2/2 LPs, were positive. The data suggest that development of LSCC should be related to the HPV infection, and HPV may be one of the inducers of LSCC.


Spectrum of autoimmune responses in systemic vasculitis. Growing evidence supports the inclusion of primary forms of systemic vasculitis within the category of autoimmune diseases. Thus their association with a family of autoantibodies, related to their specificity or neutrophil cytoplasm antigens, has been identified and the role these autoantibodies play in pathogenesis is now being explored. Different members of the autoantibody family have now been characterised in terms of their molecular specificity and the isotype of immunoglobulin of which they are composed. These properties may be a useful adjunct in the classification of systemic vasculitis (see table I). It is the purpose of this review to put a perspective on the specificity and pathogenicity of autoantibodies found in patients with systemic vasculitis and to outline new strategies for the treatment of these conditions, based on the involvement of autoimmune mechanisms in the development of these disorders.


Calvarial reconstruction in baboons with porous hydroxyapatite. The growth of bone into porous hydroxyapatite implanted in nonhealing calvarial defects of adult baboons is evaluated. Seventy-two anterior and posterior calvarial defects, 25 mm in diameter, were prepared in 24 adult male baboons (Papio ursinus). In each animal, one defect was implanted with a disc of porous hydroxyapatite obtained after hydrothermal conversion of the calcium carbonate exoskeleton of coral (genus Goniopora); pores averaged 600 microns in diameter, pore interconnections averaged 260 microns in diameter, and the void fraction was 70% (Interpore 500). Another defect was grafted with autogenous corticocancellous bone harvested from the iliac crest. The third defect was left ungrafted and used as a control. Histology and histomorphometry on nondecalcified and decalcified sections prepared from specimens harvested at 3, 6, and 9 months after surgery showed that substantial bone growth had occurred in the hydroxyapatite implants (p < 0.01 vs bone grafts at 3 mos), culminating in complete penetration of bone within the tridimensional porous spaces. Anterior untreated defects showed greater amounts of bone when compared to posterior defects. The extent of bone growth in hydroxyapatite implants, however, appeared to be independent of the site of surgical implantation. This finding suggests that bone deposition within the porous spaces was a function of the implanted matrix rather than the healing potential of specific regions within the calvaria. Of concern was the presence of nonunions at the hydroxyapatite/calvarial interfaces, despite the often extensive bone deposition within the center of the hydroxyapatite implants. Further studies are required to identify the reasons for nonunion and to evaluate the mechanical performance of the bone-porous matrix complex.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of reactive oxygen species on the erythrocyte calcium-pump function in protein-energy malnutrition. The presence of detectable amounts of non-heme iron in erythrocyte ghost membranes have been postulated to lead to the initiation of membrane lipid peroxidation and the attendant perturbation of membrane functions. We have investigated the presence of non-heme iron and endogenous products of lipid peroxidation in erythrocyte membranes of normal and kwashiorkor (KWA) subjects and assessed the susceptibility of the membranes to exogenously generated reactive oxygen species. The modulation of the basal and calmodulin-stimulated calcium-pumping activity of these membranes by reactive oxygen species was also assessed. The results show the presence of significant amounts of non-heme iron and endogenous free radical reaction products in the red cell membranes of KWA subjects compared with that of normal children. Estimation of the extent of lipid peroxidation in the presence of exogenously generated reactive oxygen species further revealed that erythrocyte ghost membranes of KWA subjects are more susceptible to oxidative stress than those of normal individuals. Although both the basal and calmodulin-stimulated activities of the membrane-bound Ca(2+)-pump enzyme in normal and KWA subjects were inhibited by oxygen-free radicals, the erythrocyte enzyme in KWA subjects showed higher susceptibility to inhibition by oxygen free radicals than that of normal individuals. We propose that the reduced erythrocyte calcium-pump function in KWA is not unconnected with excessive generation of reactive oxygen species.


Pinealectomy but not melatonin supplementation affects the diurnal variations in 125I-melatonin binding sites in the rat brain. The distribution of melatonin binding sites in synaptosomal preparations from five brain areas of sham-operated and pinealectomized young male rats (maintained in a 14 h light: 10 h darkness cycle; lights on at 5.00 h) was recorded at 10.00, 18.00 and 24.00 h, 18 days after surgery, using 125I-melatonin as a probe. The densities of 125I-melatonin binding sites in the medulla-pons, hippocampus and hypothalamus of the pinealectomized rats, exhibited clear diurnal variations. However, the densities of binding sites in these brain areas at 18.00 h were lower in the pinealectomized animals than at the other times of day tested, whereas in the sham-operated controls, the binding at 18.00 h was higher than at the other times of day. No diurnal variations were evident in the midbrain and cerebellum of the pinealectomized animals. The apparent affinities of the binding sites toward the ligand in the various brain areas were similar in the pinealectomized and sham-operated animals and did not significantly vary at any of the times recorded. Oral supplementation of melatonin to the rats via drinking water had no effect on the diurnal variations in 125I-melatonin binding in the pinealectomized rat brain. The results indicate that the diurnal variations in 125I-melatonin binding sites in the rat brain are not generated by the pineal but are affected by removal of the gland.


[Interaction of hypochlorite with oxyhemoglobin leads to liberation of iron in a catalytically active form]. Interaction of hemoglobin with hypochlorite (OCI-) induces changes in hemoglobin absorption spectra resulting in Soret band decrease and shift similar to those observed under the action of hydrogen peroxide (H2O2). Hemoglobin decomposition is accompanied by free iron release, as estimated by coloured iron-phenanthroline complex formation. The released iron is catalytically active: the incubation of hemoglobin with H2O2, OCl- or activated neutrophils increases the intensity of H2O2-dependent chemiluminescence of hemoglobin. In both reactions OCl- was more efficient than H2O2. These results show that hemoglobin can serve as a source of catalytically active ("free") iron in the reaction with OCl- and with H2O2.


The simultaneous exposition of galactose and mannose-specific receptors on rat liver macrophages is developmentally regulated. We studied the simultaneous binding of galactose and mannose-exposing ligands in sinusoidal rat liver cells during development and aging. The galactose-specific receptors were visualized using 17 nm diameter colloidal gold particles coupled with Lactosylated bovine serum albumine (LacBSA), while mannose-specific receptors were localized by means of 5 nm diameter particles adsorbed with mannan. We observed the presence of four different classes of Kupffer cells in relation to the ligands bound. The percentage of each group of Kupffer cells varied in relation to the age of the subject from which the sample was taken. There were few double-labelled cells in the livers from newborn rats, with numbers increasing with age to adulthood, and decreasing again in the older animals. Cells without labelling were in the majority after birth, but they decreased in number up to adulthood and increased again during subsequent aging. The numbers of single-labelled cells did not change significantly during liver maturation. We hypothesize that the exposition of galactose and mannose-specific receptorial systems is regulated by developmental conditions.


The determination of pentavalent antimony in sodium stibogluconate in a pharmaceutical formulation by flow injection analysis. A flow injection analysis (FIA) procedure is described for the determination of pentavalent antimony (Sb5+) in the drug, sodium stibogluconate, in a parenteral pharmaceutical formulation. The sample solution is injected directly into a carrier stream of iodide ion which is then mixed with an acid stream in situ. Sb5+ is determined by the redox reaction with acidified iodide to liberate iodine, which is monitored spectrophotometrically at 350 nm. The closed conditions prevent interference from atmospheric oxygen and the rapid reaction time assists in minimizing interference from side reactions. The use of tartaric acid as a solvent for sample and standard solutions ensures obedience of Beer's law over the Sb5+ concentration range 0.01-0.2% (w/v). The method is specific for the higher oxidation state in an ionic mixture of Sb5+ and Sb3+, and has been fully validated for use in a pharmaceutical preparation. Assuming absence of matrix interference it is applicable to Sb5+ from other sources and should be applicable to other reducible ionic species.


Determination of disodium clodronate in bulk material and pharmaceuticals by ion chromatography with post-column derivatization. A validated ion chromatographic method for the determination of disodium clodronate in bulk material and pharmaceuticals is described. Separations are performed on a poly(styrene-divinylbenzene) copolymer column using 40 mM nitric acid as mobile phase at a flow rate of 0.5 ml min-1. The analyte is detected by UV absorption at 300 nm after post-column derivatization with acidic iron(III) solution (0.25 ml min-1). The proposed ion chromatographic method is validated in terms of selectivity, precision, linearity, accuracy and ruggedness.


The effects of two air-powder abrasive prophylaxis systems on the surface of machined titanium: a pilot study. This in vitro pilot project compared the effect of two air-abrasive prophylaxis systems on the surface of machined titanium. Single Brånemark titanium abutment cylinders were exposed to the Prophy-Jet and Microprophy systems for 90 seconds each. Both of the test cylinders were compared with an untreated control cylinder by scanning electron microscopy. Machining marks were completely removed by the Prophy-Jet and only partially removed by the Microprophy. Both of the resultant surfaces appeared to be smoother and thus may be more resistant to plaque formation. A rationale for the removal of machining marks is presented, although the reason for the difference in removal by the two systems is unclear. The prophylaxis cleaning powders were also examined by scanning electron microscopy and exhibited similar particle dimensions and morphology. A noncrystalline deposit was observed on the surface of the abutment cylinder exposed to the Microprophy. Energy dispersive spectroscopy analysis revealed that the deposit consisted almost entirely of sodium. Further investigation of the deposit is needed.


[The response of the airways in asthmatic patients to kinin inhalations define a type-B2 receptor profile]. Kinins (i.e. bradykinin, kallidin and [desArg9]-bradykinin) are vasoactive oligopeptides which may contribute as mediators in the pathogenesis of asthma by interacting with specific receptors designated as B1 and B2. The aim of the present study was to investigate the airway response to inhaled histamine, bradykinin, kallidin and [desArg9]-bradykinin in normal and asthmatic subjects. Changes in airway caliber were followed as FEV1 (forced expiratory volume in 1 sec) and as Vp30 (maximum expiratory flow at 30% of the vital capacity). Neither histamine, bradykinin, kallidin nor [desArg9]-bradykinin had any measurable bronchoconstrictor effect in the normal subjects. However, in the asthmatic subjects, histamine, bradykinin and kallidin, but not [desArg9]-bradykinin, produced prompt concentration-related bronchoconstriction. The geometric mean PC20FEV1 (provocation-concentration of inhaled agonist reducing the FEV1 by 20% from baseline) values were 0.027, 0.082 and 0.44 mg/mL for bradykinin, kallidin and histamine respectively. Because bradykinin and kallidin are agonists of B2 receptors and [desArg9]-bradykinin is an agonist for B1 receptors, our data suggest that asthmatic, but not normal, airways are hyperresponsive to kinins and that this potent bronchoconstrictor response is due to a specific pharmacologic effect compatible with the stimulation of B2 receptors.


[Immunological effects of live attenuated hepatitis A vaccine]. 11,451 health persons were inoculated with live attenuated hepatitis A vaccine (H2 strain) in 3 batches. Among them, 785 were tested for anti-HAV antibody response by ELISA. The results showed that H2-strain vaccine got a quick antibody response (2-5 weeks) and a high seroconversion rate (92.9%). Also, the titers of antibodies to HAV ranged from 1:2 to 1:32. The seroconversion of the first 12 recipients lasted for 3 years. The primary epidemiological studies demonstrated that H2-strain vaccine was effective in preventing hepatitis A. The relation between the positive rate of antibody and different dosage showed that 10(6.5)TCID50 could get a highest seroconversion rate (100%).


Effects of superoxide anions on red cell deformability and membrane proteins. The effect of superoxide anions (O2-) on red blood cells (RBC) deformability and membrane proteins was investigated using hypoxanthine-xanthine oxidase system. Exposure of RBC to O2- caused a marked decrease in RBC deformability with a concomitant increase in cell volume and shape changes. The RBC exposed to O2- also displayed pronounced degradation of membrane proteins such as band 3 protein and spectrin; new bands of low molecular weight products appeared as the original membrane proteins tended to diminish, without the appearance of high molecular weight products. Since the membrane proteins are involved in processes regulating membrane properties such as permeability and viscoelasticity, the decreased deformability induced by O2- may be attributable to changes in membrane proteins. Interestingly, resealed ghosts exposed to O2- did not show any significant change in membrane proteins, which suggests the existence of further generation of O2- and subsequent production of other active oxygen species mediated by O2(-)-initiated autoxidation of hemoglobin in intact RBC. Furthermore, electrophoretic analysis suggested that active oxygens increased the endogenous proteolytic susceptibility of RBC. In conclusion, a close linkage was suggested between RBC deformability and the membrane proteins.


[Circulating autoantibodies against beta-adrenergic receptors in patients with heart diseases]. Patients with different heart diseases, dilated cardiomyopathy, valvular heart disease, hypertension, ischemic heart disease or myocarditis showed manifestations of autoimmunity and down-regulation of beta-adrenergic receptors. Autoantibodies against beta-adrenergic receptors in these patients were detected with radioligand binding inhibition assay. The results suggested that the down-regulation of cardiac beta-adrenergic receptors in these patients may be mediated by autoimmunity. Autoantibodies against beta-adrenergic receptor were not related to any specific heart diseases, but to the severity of heart failure irrespective of its etiology. The significance of these autoantibodies in heart failure was discussed.


[Dot immunobinding assay in detection of IgE-type rheumatoid factor]. The sera from 86 patients with autoimmune diseases, 31 atopic and 65 healthy blood donors were detected for IgE type rheumatoid factor (IgE-RF) by using anti-human IgE monoclonal antibody and dot immunobinding assay. The positive frequency and mean value of IgE-RF in patients with autoimmune diseases were significantly higher than those in the other groups. IgE-RF was positive in 19/37 cases of rheumatoid arthritis (RA), 6/18 systemic lupus erythematosus and 13/31 primary Sjogren's syndrome, with mean values of 23.1, 12.1 and 26.9 ng/ml respectively. 75% RA patients with IgM-RF (+) had IgE-RF, with a mean value of 27.0 ng/ml, whereas 33% of those with IgM-RF (-) had IgE-RF with a mean value of 16.5 ng/ml. In contrast, only 2/31 atopic and 6/65 healthy blood donors had positive IgE-RF with mean values of 3.7 and 6.1 ng/ml respectively. Hence, IgE-RF is helpful not only in raising the sensitivity of RA diagnosis, but also in distinguishing autoimmune diseases from allergy.


[Epidermal growth factor and its receptor in the renal tissue of patients with acute renal failure and normal persons]. Epidermal growth factor (EGF) and its receptor (EGF-R) were identified by immunohistochemical method (4 layer PAP) in the renal tissue specimens obtained from 11 normal kidneys and 17 cases of acute renal failure (ARF). The quantitative EGF and EGF-R in the tissue were expressed as positive tubules per mm2. The amount of EGF and EGF-R in renal tissue was markedly increased in cases of ARF than in normal persons. Positive EGF staining was present only in distal tubules, but absent from glomeruli in normal persons. In ARF patients EGF was identified in both distal and proximal tubules. It was also seen in the glomerular mesangial area in 2 cases complicated by mesangial proliferative GN. However, no obvious difference was noted in the distribution of EGF-R between normal and ARF specimens. The amount of EGF in the renal tissue varied with the morphological changes in tubules and the time when the renal biopsy was being taken. The amount of EGF was large in those with a Scr value less than 2.5mg/dl There was a more profound expression of EGF during the recovery from ARF.


[Survey of bacterial resistance in Shanghai district]. The data on bacterial resistance monitoring of 5,457 clinical isolates in 1990 against 21 antimicrobial agents showed that most isolates were highly resistant to commonly used antimicrobial agents, such as penicillin, ampicillin, erythromycin, chloramphenicol, tetracycline and gentamicin, with a resistant rate of around 50% or more. Strains resistant to some newer agents, such as cefotaxime, ceftazidime, amikacin and norfloxacin, have appeared and are increasing in number. Multi-resistant strains, i.e. strains resistant to five more antimicrobial agents existed in various bacterial species. Vancomycin was the most active antibiotic against methicillin-resistant staphylococci and no resistant strains were found. Strains of Salmonella and Shigella spp. were still very sensitive to the antimicrobial agents tested, with a sensitive rate of more than 90%. 80% of 765 gentamicin-resistant strains of Enterobacteriaceae were still sensitive to amikacin.


[Magnetic stimulation motor evoked potential in patients with Guillain-Barré syndrome]. In 27 patients with Guillain-Barré syndrome (GBS), motor evoked potentials (MEP) were compared with electromyography (EMG), nervous conduction velocity (NCV), F wave and somatosensory evoked potentials (SEP). The positive rate of MEP was 74%, and 83% of EMG. The findings of MEP were consistent with pathological changes of demyelination and clinical distribution of muscle weakness. A good correlation between abnormal MEP and degree of muscle weakness (P < 0.01) was noted. MEP should be regarded as an objective index for diagnosis, differential diagnosis, followup and effect of therapy. MEP is painless and can be done easily and repeated.


[Epidermal growth factor for enhancing DNA synthesis of hepatocytes and its protecting effect on animals with liver injury]. Epidermal growth factor (EGF) was purified chromatographically from mice submaxillary glands, and its activity and electrophoretic pureness were identified. The effect of EGF, glucagon-insulin (G-Ins) and EGF-glucagon-insulin mixture(EGF-G-Ins) on stimulation of DNA synthesis in primary cultures of rat hepatocytes and their protective effect on mice with liver injury were investigated. The results showed that G-Ins had significant effect on DNA synthesis in primary hepatocyte culture; EGF showed more significant effect compared with G-Ins; and EGF-G-Ins had the best effect. EGF-G-Ins could increase the survival rate and improve the repair of injured hepatocytes in mice treated with D-galactosamine. Although EGF and G-Ins could reduce the degree of hepatic injury, they did not elevate the survival rate. The present study provided some information on clinical therapy with EGF-G-Ins in patients with fulminant hepatic failure.


[A research for screening anti-hepatitis B virus drugs with the 2.2.15 cell line]. In this paper, 43 drugs have been screened and evaluated for their ability of anti-hepatitis B virus by way of detecting in the cell culture medium the HBsAG and HBeAG secreted from the 2.2.15 cell line--a cell line derived from the Hep-G2 transfected with cloned HBV DNA. In addition, the MTT colorimetric assay is used in the process for monitoring the cytotoxic effects. The result showed that five drugs, including anti-HBV-I, anti-HBV-II, Radix Astragali Co., Rhizoma Curculiginis Co. and 518BII-A-6 are promising anti-HBV drugs.


[Treponemal pallidum specific 19S-IgM haemagglutination test and its clinical significance]. 74 samples of 53 cases were detected with 19 S-IgM-TPHA. The sensitivity was 97.37% and the specificity was more than 99%. 19S-IgM-TPHA was not interfered by rheumatoid factor. It was used to define congenital syphilis, to determine the necessity of treatment and reinfection, to diagnose syphilis without lesion and detailed case history as well as neurosyphilis. The results of 19S-IgM-TPHA were not consistent with those of routine screen and definite tests (Kappa < 0.4), so it can not be replaced by other methods.


[Effect of placental transferrin receptors on iron nutritional state of normal full-term gravidas and their newborns]. Sixty full-term gravidas were divided into four groups, normal insidious iron deficiency, mild iron deficient anemia and moderate iron deficient anemia, according to their iron nutritional state determined by the measurement of hematologic and iron biochemical indexes. The iron nutritional state of newborns in each group and the level and the affinity of transferrin receptor in placenta were also studied. The iron nutritional state of newborns was found to decrease mildly along with the decrease of their mothers' iron nutritional state, especially the decrease of serum ferritin, but the differences were not statistically significant. It was suggested that the iron nutritional state of newborns was relatively normal although their mothers were in severe iron deficiency. The differences of transferrin receptor levels were significant among the four groups, the mild IDA group had the highest level of transferrin receptor, which was 1.68 times of normal group and 1.77 times of moderate IDA group. The differences between each two groups were significant. The differences of dissociate constant (Kd) of transferrin receptor were not significant among the four groups, indicating that the iron metabolism between mothers and their babies was regulated not by changing the affinity of transferrin binding to its receptor but by changing the numbers of transferrin receptor to maintain the relative stableness of newborns iron nutritional state.


[The inhibitory effect of high energy shock waves on bladder tumor cell line BIU-87 in vitro]. The cytotoxic effect of high energy shock waves (HESW) on bladder tumor cell line BIU-87 in vitro was evaluated by using the lithotripter Dornier HM3 with voltage 15 kV, impulsation 0, 400 and 800 respectively, water bath 35-37 degrees C and impulse rate 60/min. Cell viability and clonogenicity were decreased, but the cell doubling times were prolonged with the increasing number of HESW impulsation. Both cell growth curves and 3H-thymidine incorporation assay showed the temporary inhibition on the growth of tumor cells, which were viable after HESW. After a period of time, these tumor cells would recover their normal growth ability. For the group received 800 HESW impulses, the first passage of cell growth curve decreased temporarily 24 hours after HESW impulsation. These results indicate that HESW do show a cytotoxic effect on tumor cells in vitro, including acute and chronic lethal effect and temporary growth inhibition. The cytotoxic effect of HESW is positively proportional to the number of impulsation at the condition of definite working voltage.


[High energy shockwave-induced acute changes in renal function]. Attempting to understand the effects of HESW on renal function, we studied prospectively 40 patients with nephrolithiasis in 4 groups, using different number of pulsation and the same voltage to identify different effects. Stone burdens and position were similar in these groups. Each group received 1,500, 2,000, 2,500, or 3,000 pulses respectively at 12.5 kV from JT-3 lithotripter. In all groups, the levels of urinary NAG, beta 2MG, ALB and serum beta 2MG were significantly increased at day 1-3 after ESWL (P < 0.001), and then decreased to the levels of pre-ESWL except serum beta 2MG and urinary NAG levels of group C and D at day 7 after ESWL, which were significantly higher (P < 0.05) than those of pre-ESWL. There was significant correlation between either urinary NAG (r = 0.977, P < 0.05) or urinary beta 2MG (r = 0.933, P < 0.001) and the number of pulses at day 3 post-ESWL. In addition, there was a significant difference in urinary NAG levels between group D and group A, B or C at day 3 post-ESWL, and the same was true in urinary beta 2MG levels between group C or D and group A or B. These findings suggested that shock wave induced acute changes in renal function and transient renal tubular damages, and that the tubular damages might last longer more than 7 days, although these functional changes recovered within one week. The changes were related to the energy levels of shock wave, and the degree of renal damage would increase when the energy level was above 12.5 kV x 2,500 pulses.


[Long-term effect of extracorporeal shockwave lithotripsy on kidney. A preliminary study]. We studied 139 patients with upper urinary tract calculi who were treated by JT-ESWL-I and III type extra corporeal shock wave lithotripter (ESWL) for an average of 4 years and 11 months. These patients aged below 45 had no history or family history of hypertension. Hypertension was found in 1 patient (0.7%) and elevated diastolic pressure in 6 (4.3%). The recurrence rate of calculi was 5%. Before and after ESWL, Bun, Cr, B2-mG (of blood and urine), activity of blood plasma renin AT-II, -GT, NAG, mucoprotein of plasma, etc were also determined. The differences in their values were not significant. We concluded that if the indications for ESWL are strictly considered including patient selection and number of wave pulsation and energy, ESWL is still an ideal method of treatment.


[Transurethral microwave radiation for bladder neoplasm]. 42 patients with bladder neoplasm were treated effectively by transurethral microwave radiation in our hospital from March, 1990 to October 1991. No serious complication was found after this treatment and its long-term result was better than others. The principle of this method is to use the biological availability of microwave to heat the neoplasm and to evoke the cellular immunity system to kill tumor cells, because tumor cells is more likely to be killed than normal tissue cell by heat. Generally, tumor cells would be killed when the temperature is higher than 42 degrees C. The protection against microwave during the treatment was also discussed.


[Lupus erythematosus in pregnancy. A prospective study]. The maternal and fetal outcome in 20 patients with lupus erythematosus in pregnancy was studied prospectively. The conditions of the patients were: remission in 4 patients, inactive (controlled by 10 mg prednisone for at least 6 months) 11, active 5 (active disease while pregnancy 3 and first onset and diagnosis during pregnancy 2). There were no maternal death, fetal loss or still birth in full pregnancy course. But the intrauterine malnutrition, low placenta weight and low body weight of newborns were significantly associated with active maternal disease (P < 0.001, P < 0.05). We conclude that (1) family planning pregnancy should be planned in a period while the active disease has been well controlled with a small dose of corticosteroids for at least 6 months. (2) carefully monitoring of the disease activity at intra- and post-partum is essential to lupus patient for a successful birth.


[Transcatheter hepatic arteriography combined with ultrasound in the diagnosis of space occupying lesions]. Super-selective hepatic arteriography was performed in 40 patients with liver neoplasma by injecting H2O2. The final diagnosis were as follows: 23 hepatic cases of carcinoma, 14 patients had cavernous hemangioma, 1 case of hepato-cirrhosis and 2 cases excluding neoplasma. According the contrast, the entrance of H2O2 into neoplasma could be divided into four types: rapid-filling, middle speed-filling, low speed-filling, enclostic but no entering. The acoustic contrast technique, pathological basis, and qualitative diagnosis were also discussed.


[Antibodies to extractable nuclear antigen polypeptides: detection and its clinical significance]. Using immunoblotting technique (IBT) and saline extracts of rabbit thymus powder as antigen, we detected seven autoantibodies i.e. anti-Sm, anti-RNP, anti-SSA, anti-SSB, anti-Scl-70, anti-Jo-1 and anti-ribosome with only a strip of blot. Comparison of IBT and counter-immunoelectrophoresis (CIE) showed that the antibodies against Sm in SLE could be detected more sensitively by IBT than by CIE (P < 0.01). And the antibodies against ribosome by IBT were much specific for SLE (P < 0.01). So the detection of antibodies against ENA polypeptides by IBT was helpful in the diagnosis of SLE. In rheumatic diseases, the antibodies against SSB could be detected more sensitively by IBT than by CIE (P < 0.05), while the antibodies against SSA could be detected more sensitively by CIE than by IBT (P < 0.01). The antibodies against Scl-70 and Jo-1 by IBT were much specific for PSS and PM/DM respectively (P < 0.01).


[Terminal deoxynucleotidyl transferase in various immunophenotypic cells in leukemia]. The staining results of terminal deoxynucleotidyl transferase (TdT) in leukemic cells of peripheral blood or bone marrow in 55 patients showed that 22 (40%) patients were found positive for TdT staining: 17 patients with acute lymphoblastic leukemia (ALL), 2 acute non-lymphocytic leukemia (ANLL) and 3 hybrid acute leukemia (HAL). The positive rate of ALL (17/21) was significantly higher than that of ANLL (2/21, P < 0.005). The leukemic cells of HLA-DR+CD19+TdT- were demonstrated to express some of myeloid restricted antigens (HI98, HIM 4, HIM 5) (3/3), which suggested that these cases were leukemias of myeloid origin. None of the 8 patients with M 0/M 1 ANLL was positively stained for the enzyme. Our results showed that TdT detection is of great value in the diagnosis and differentiation of both poorly differentiated and hybrid acute leukemia.


[Correlation of intra-erythrocytic Na, K, Ca, Mg contents and ATPase activity with dilated cardiomyopathy]. Intra-erythrocytic contents of Na, K, Ca, Mg and activities of Na-K-ATPase (Na-pump) and Ca-Mg-ATPase (Ca-pump) were measured in 30 cases of dilated cardiomyopathy (DCM). Effects of treatment on these variables were simultaneously observed. The results showed that intra-erythrocytic Na, Ca contents increased and K, Mg contents decreased. That erythrocyte membrane Na-pump and Ca-pump were remarkably lower in DCM than those in the controls. Red cell cations and ATPase activities returned to normal following the recovery of cardiac function. It is suggested that abnormalities of intracellular cations and cell membrane cation transfer may play an important role in the mechanism of DCM.


[Doppler echocardiography for measuring the right ventricular diastolic function]. To noninvasively measure the right ventricular diastolic function, continuous wave Doppler echocardiography was used in 31 patients with tricuspid regurgitation (TR). The right ventricular pressure (RVP) curves were analyzed mathematically, showing that RVP declined exponentially after maximum-dP/dt. The instantaneous RVPs during isovolumetric relaxation were measured from RVP curves and also calculated by using exponential equations. There were high correlations between measured and calculated RVPs (r = 0.83-0.98). The tricuspid regurgitant velocities were recorded by CWD and converted into pressure gradients (PG) using a simplified Bornoulli equation. The instantaneous RVPs were then obtained by adding an assumed right atrial pressure to PGs. The maximum-dP/dt, the instantaneous RVP at the time of -dP/dt max (PO) and the time constant of right ventricular relaxation (T) were calculated from both RVP curves and Doppler spectra. The results showed that there were high correlations of -dP/dt max, PO and T measured by two techniques (r = 0.95, 0.98 and 0.92). We conclude that T can be used to quantitate right ventricular myocardial relaxation, and CWD offers a new technique for noninvasively measuring T in patients with TR.


[Hemodynamic effects of amrinone on cardiopulmonary decompensated chronic cor pulmonale]. Amrinone was given intravenously combined with the routine treatment to 10 patients with late chronic cor pulmonale in exacerbation stage. The initial loading dose was 1.5-2 mg/kg followed by the maintenance dose of 15 micrograms/kg. min for 6 hours. Hemodynamic and blood gas monitoring showed that pulmonary artery pressure, pulmonary vascular resistance index, right atrial pressure and pulmonary capillary wedge pressure decreased significantly and maintained the level markedly below baseline after amrinone administration (the extents decreased were 0.53-1.1 kPa, 80-140 dyn.s.m2.cm-5, 0.27-0.40 kPa and 0.40-0.67 kPa respectively. P < 0.05 for each parameter) and that cardiac index and stroke volume index increased identically (the extents increased were 0.3-0.6 L/min.m2 and 2-4 ml/beat.m2 respectively, P < 0.05 for each parameter). There were no significant changes in systemic artery pressure, arterial blood gas analysis and blood platelet count after amrinone administration. We suggest that amrinone may be effective in the treatment of cardio-pulmonary decompensated chronic cor pulmonale.


[Brain damage in asphyxiated newborn infants: early diagnosis. A follow-up study]. Cranial computerized tomographic (CT) scan and serum creatine phosphokinase-BB(CPK-BB) determination were done in 85 asphyxiated newborn infants, in some of them, long-term follow-up study including neurodevelopmental assessment and CT scan were also carried out. The types of early brain damage after perinatal asphyxia included intraparenchymal hemorrhage (IPH), periventricular-intraventricular hemorrhage (PV-IVH), subarachnoid hemorrhage (SAH) and brain edema (BE). Serum CPK-BB levels were elevated within 24 hours after the events of asphyxia, and correlated to the severity of brain damage. These findings were helpful in the early diagnosis of brain damage and estimating its severity and prognosis. The types and severity of brain damage were obviously related to prognosis. The prognosis of infants after SAH and BE were better than those of infants with PV-IVH, and infants with IPH showed the poor results. Follow-up CT scans were performed in 30 infants with brain damage. Abnormal changes included ventricular sulcus dilation, leukomalacia, cyst formation (pseudocyst) and porencephalic cavitation (porencephaly).


[Clinical trial on termination of early pregnancy by using dl-15 methyl prostaglandin F2 alpha methyl ester in combination with testosterone propionate. Report of 244 cases]. 244 subjects with amenorrhea < or = 56 days were given intramuscular injection of testosterone propionate 100 mg per day for 3 days, and in the morning of day 4, 1 mg dl-15 methyl prostaglandin F2 alpha methyl ester (PG05) suppository was inserted in the posterior fornix of the vagina every 2 hours up to the pass-out of the villi and sac for utmost 5 mg. At the same time 1-2 tablets of diphenyloxylate Co. were taken. The complete-abortion rate was 83% (191/231), the incomplete abortion rate was 13% (30/231), the total efficacy rate was 96% (221/231), and the failure rate was 4% (10/231).


[Demethyl-vancomycin in treatment of resistant staphylococcal infections. A clinical evaluation]. 77 patients with serious resistant staphylococcal infections, including septicemia, lower respiratory infection, intraabdominal infection, skin and soft tissue infections and urinary tract infection, were treated with demethyl-vancomycin. 82% of the organisms were methicillin-resistant. Most of the patients had severe underlying diseases and were immunocompromised hosts. The infections were serious. The clinical efficacy rate was 73% and the bacteria clearance rate 68%. Mild adverse reactions happened in 11% of the patients and no obvious nephrotoxicity was noted. MIC90 of demethyl-vancomycin against staph. aureus was 2 micrograms/ml. All isolates in this study were highly susceptible to the drug. Demethyl-vancomycin was found more active against staphylococcus than the other 16 antibacterial agents, which are commonly used in this country. The indication and the use of the drug were discussed.


[Effect of captopril on human myocardial ACE-inhibition: neurohumoral variations during coronary sinus cardioplegia. Cardioprotective effect during aortocoronary bypass]. The present study is aimed at checking effects exerted by captopril on human myocardial ACE system (ACE-T) as well as the role played by tissue ACE-inhibition in reducing reperfusion damage. A human experimental model was used during cardioplegia due to aortocoronary bypass graft (CABG): 54 patients with coronary artery disease affecting 3 vessels having suffered from acute anterior myocardial infarction, homogeneous as far as ejection fraction (35-55%), number of grafts (3), clamping time, age and sex, were randomised in a double blind experiment, and were given captopril or placebo: 4 mg/l captopril were mixed into the cardioplegic solution with blood according to Buckberg, 8 blood samples were obtained from each patient and norepinephrine, epinephrine were assayed using an HPLC technique. Angiotensin 1 was assayed by RIA. CK was assayed as well (U/L). Blood samples were taken during CABG: pre-pump sample; pump sample; pump preclamping sample; coronary sinus sample; warm reperfusion sample; coronary sinus during warm reperfusion; after clamping sample; after cannulation. captopril group (29 patients): angiotensin 1:8.15; 6.9; 7.45; 8.66; 8.93; 8.70; 9.07; 9.40 versus placebo: 7.09; 7.43; 7.80; 9.31; 9.01; 8.35; 8.85; 8.07 mcg (all NS). Noradrenaline: captopril group: 359; 404; 329; 282; 263; 216; 310; 337 versus placebo: 439; 520; 499; 469; 526; 566; 501; 443 pg (p < 0.001). CK, captopril group: 79.9; 95.1; 100.8; 94.3; 104.2; 94.7; 108.4; 108 versus placebo: 76.2; 120.2; 135.5; 203; 225; 272; 247; 228.7 U/L (p < 0.01). Epinephrine values showed no significant difference between the 2 groups. Norepinephrine and CK decrease as well as angiotensin I increase in treated patients as compared to controls suggest some effect exerted by captopril ACE-T and its capability of reducing reperfusion damage and recommend its use for heart protection during CABG.


Angioedema complicating lisinopril therapy. A case of angioedema induced by antihypertensive therapy with lisinopril is presented. The patient was a 70 year old black woman, with a history of hypertension for 15 years. The patient presented with acute onset of swelling involving the oro-facial region and respiratory distress after ingestion of three doses of lisinopril over a three day period. A clinical diagnosis of drug induced angioedema was made based on clinical presentation. The patient was treated with diphenhydramine, 50 mg intravenously, and hydrocortisone 100 mg every eight hours with resolution of her symptoms over a 24 hour period. Angioedema should be recognized as a possible life threatening complication of therapy with lisinopril, and other angiotensin converting enzyme (ACE) inhibitors. This usually responds to therapy with antihistamines and steroids if recognized early.


[Sister chromatid exchanges in lymphocytes of patients with gynecological malignancies]. The frequency of SCE in the peripheral blood lymphocytes of 30 patients with gynecological malignancies was investigated. The frequency of SCE was found to be 5.37 +/- 0.44, 5.30 +/- 0.35 and 3.76 +/- 0.38 respectively in the patients before and after chemotherapy and in the control cases. The SCE values deviated significantly from those of control, and were not related to the staging of the tumor. The study suggested the possibility of utilizing SCE as a cytogenetic indicator in some malignant tumors.


[Insight into the mechanism of torsade de pointes (TDP) induced by cesium chloride]. Standard intracellular potential recording technique was used in isolated guinea pig ventricular muscle and monophasic action potential (MAP) recording technique was performed on both the epicardium and endocardium of dog hearts in vivo. The results showed that CsCl lengthened action potential durations of ventricular muscle as well as QT intervals of dog hearts. The incidence of early after depolarization (EAD) induction was 71.4%. Ventricular tachycardia induced by CsCl manifested as typical TdP in 44.4% with EAD shown on epicardial MAP recording. This suggested that EAD and triggered activity might be an important mechanism of TdP. As verapamil could depress the induction of EAD, we believe that Ca++ inward currents was responsible for the induction of EAD and TdP by CsCl.


[Study of toxication of toxoflavin from Pseudomonas cocovenenans to immunocyte and detoxication]. In order to study the toxication of toxoflavin produced by P. cocovenenans to immunocytes and detoxication, the effects of toxoflavin on rabbit blood culture in the presence of PHA and detoxication function of seven drugs towards this system have been observed by 3H-TdR incorporation assay. The more toxoflavin used, the more reduction of cpm value. The results indicated that toxoflavin in 0.5 microgram/ml or more suppressed the growth of cells to even still lower than that of control in which PHA was absent; and when 0.1 microgram/ml of toxoflavin were used, the cpm value began to rise again. The effects of toxoflavin on lymphocyte and other blood cells were studied under the microscope and the naked-eye. The detoxication effects of the seven drugs on toxoflavin were observed and shown that SOD, GSH, Cyt. C and VC were most effective in protecting the lymphocytes from poisoning.


[The mechanism of the intestinal phase of pancreatic secretion]. Literature references and author's own results on the role of the pancreatic secret in the intestinal phase of pancreatic secretion, are reviewed. Particular attention was paid to formation of correcting effects of the duodenum upon pancreatic secretion in respect to the ratio of solubilised and absorbed pancreatic enzymes, enzymes, substrates and the products of their hydrolysis in the duodenal chyme.


[Peptides in the regulation of digestive functions in fish]. The effect of exogenous peptides (angiotensin II, neurotensin, thyroliberin, pentagastrin, cholecystokinin, insulin) on the motor and myoelectrical activity of the sea fish digestive tract and their food behaviour was found to depend on the technique of administration and the fish species. Some peptides could change the intensity of consumption in forage fish. Specific receptors for peptides seem to exist in the fish.


[The effect of heat exposure on the circadian rhythm of the enzymatic activity in different sections of the rat small intestine]. Two-hour heat exposure (40-41 degrees C) was shown to change the circadian rhythms of enteral gamma-amylase, maltase and sucrase activities. The character and obviousness of the changes depended on the type of enzyme activity and the portion of the small intestine. The heat exposure did not change daily mean levels of enzymatic activities and their distribution along the small intestine.


[The role of lysosomal enzymes in the formation of destructive processes in the gastroduodenal area]. The paper reviews works by the author and his colleagues on the problem of pathogenesis of ulcer formation within the gastroduodenal area. The stomach enzymes active in weakly acid pH zone 4.0-7.0, were shown to play a major part in the mechanisms of ulcer formation. These enzymes are of lysosomal origin and the ulcer formation starts within the mucous-submucous layer's tissues. Destabilizing of the lysosomes and release of autoaggressive lysosomal enzymes occurs under the effect of excessive production of the biogenic amines histamine and serotonin. Adrenal hormones are also involved in the process of ulcer formation.


[The general functional characteristics of endogenous regulatory oligopeptides]. A number of physicochemical factors of the endogenous regulatory oligopeptides are considered to be the basis of their general functional properties as indicated by the phenomena of cascade regulation, morphogenesis, behaviour and biological evolution. Functional characteristics and chemical features of classical transmitters and, probably, other endogenous substances allow to consider them as the integrity of endogenous molecular regulators.


[The reactions of the viscerosensory neurons of the nucleus solitarius to stimulation of the posterior and anterior hypothalamus]. The "vagal" neurons of the solitary tract nucleus responded to stimulation of anterior and posterior hypothalamus in 78.7 and 27.6 per cent, resp., the responses being mainly of the activation type. The descending effect from the hypothalamus involved primary as well as secondary "vagal" neurons. The responses were of mono-, oligo- and polysynaptic nature.


[The previously unknown biological characteristics of the neurons of the paraventricular hypothalamic nucleus]. Structural sex-depended differences were revealed in 7 out of 10 hypothalamic paraventricular nuclei under study in DB/DB C57 BL-K56 mice. Sex-dependent differences were also found in the responses to diabetes mellitus in 6 of these nuclei. The data obtained seem to corroborate the sex dependence of the paraventricular-vagal neuronal system involved in the control of carbohydrate homeostasis.


[The mechanisms of the influence of the thalamic nuclei on the heart]. A major part of the ventromedial hypothalamus and the solitary tract nucleus in realisation of the thalamic effects on the heart, was revealed in rabbits. The polychemosensitive organisation of the thalamic neurons prompts specific functional shifts in the myocardium.


[The digestive enzymes in the gastrointestinal tract, kidney, liver and spleen in different functional states]. A wide range of digestive enzymes with different cellular location (membrane, intracellular, Lysosomal) was determined in gastrointestinal organs (stomach, duodenum, jejunum, ileum, colon) as well as in undigestive organs (liver, kidney, spleen) in normal conditions and in altered functional states (fasting, refeeding). High levels of peptidase activity was noted in undigestive organs and the colon as compared to that in the small intestine. Adaptive responses were (revealed not only for a number of membrane enzymes but also for intracellular ones including those in undigestive organs.


[The place of the gastrointestinal tract in the regulation of feeding behavior]. The paper reviews the role of neural and humoral signals associated with the gastro-intestinal tract's activity, in organisation of motivational and emotional behaviour. The author rejects the idea of insignificance of the digestive channel's role in formation of the states of hunger and satiety. Conditioned readjustment of initiating and inhibitory afferents from mechano-, osmo- and chemoreceptors of the stomach and upper portions of the intestine, are shown to play a part in formation of general and specialised appetites according to the body needs.


[The spatial distribution of mitochondrial respiration and oxidative phosphorylation along the small intestine in rats of different ages]. Growing rats were shown to have the maximum of respiratory and ATP-synthetase activities in the duodenum and proximal part of the small intestine, the minimum--in its distal part, whereas the medial part of the intestinal tube was in intermediate position. These parameters were equally distributed along the small intestine in adult rats. In old rats, the highest rate of oxygen consumption was revealed in the mitochondria of the medial part, the lower one--in the proximal part, and the lowest level was found in the duodenum and in distal part of the small intestine.


[Apolipoprotein C III binding sites (receptor) on non-parenchymal cells from rat liver]. Binding sites of apolipoprotein (apo) C III on non-parenchymal cells (NPC) isolated from rat liver were found. Apo C III was purified from delipided human plasma very low density lipoproteins. 125I-labeled-apo C III was prepared by chloramine-T method. Non-parenchymal cells were isolated from rat liver by collagenase method. Freshly isolated rat liver NPC bound 125I-labeled apo C III in a saturable way with Kd 0.1-0.55 mumol/L and Bmax 18.1-42.0 ng/ml cell protein. There were about 1.0-4.5 x 10(5) specific binding sites of apo C III on each NPC. Unlabeled apo C III, but not apo AI and B100, could inhibit these binding activities. The results demonstrate the presence of saturable and specific apo C III receptors (binding sites) on rat liver NPC, but their nature and biologic properties remain to be investigated.


[erb-B oncogene in human oral squamous cell carcinomas]. erb-B gene is an oncogene from avian leukosis virus. The carcinogenic capability of erb-B is due to the close similarity of expressional proteins to the epidermal growth factor receptor (EGFr). In this study the amplification and the over-expression of erb-B were found in human oral squamous (cell carcinomas by DNA and RNA dot blot hybridization). Results showed that erb-B was correlated with the oral epidermoid tumors but not with group of non-epidermoid tumors in the head and neck region. Southern blotting analysis showed that there were rearrangement of erb-B gene in the tumor genosomes, which is perhaps one of the ways to activate the proto-oncogene of erb-B.


Mutant of Salmonella typhimurium lacking the inhibitory function for phagosome-lysosome fusion in murine macrophages. It has recently been described that Salmonella typhimurium is capable of inhibiting phagosome-lysosome fusion in murine macrophages after ingestion. We selected a mutant of S. typhimurium lacking the phagosome-lysosome fusion inhibitory function from a collection of Tn5-insertion mutants and examined its relevance to the pathogenesis in mice. The Tn5 insertion mutant which has a defect in fusion inhibitory function was found to be significantly sensitive to the intracellular killing by murine macrophages in vitro. However, the loss of the fusion inhibitory function did not reduce the level of virulence for mice in vivo. These results demonstrated that fusion inhibition did not play a critical role in the pathogenesis of S. typhimurium although it might contribute to at least a part of the resistance against macrophage killing mechanisms.


[Serum activity of angiotensin-converting enzyme and osteocalcin levels in hyperthyroidism]. This study was carried out in order to investigate serum changes of osteocalcin (BGP) and angiotensin converting enzyme (SACE) activity in a group of patients with hyperthyroidism. We studied 20 hyperthyroid patients (F 14, M 6; age mean 37.5 +/- 16.8 years) and 13 control subjects (F 11, M 2; age mean 40.3 +/- 7.5 years). In both patients and controls we measured: FT3, FT4, T3, T4, TSH, BGP, SACE. Finally, in patients with hyperthyroidism a TRH test and functional investigations were also performed. We observed that mean SACE levels were significantly increased in hyperthyroid patients (32.06 +/- 10.3 nmol/ml/min) in respect to control subjects (14.66 +/- 3.88 nmol/ml/min) (p = 2.02 E-6). Similarly serum BGP levels were significantly increased in hyperthyroid patients (5.94 +/- 2.55 ng/ml) than in control subjects (2.89 +/- 1.58 ng/ml) (p = 5.66 E-4). There was a significant linear correlation between SACE and T4 levels (r = 0.48; p < 0.05), between serum BGP and T4 (r = 0.50; p < 0.02) and furthermore between SACE and BGP (r = 0.57; p < 0.01). In conclusion both serum BGP and SACE levels are increased in patients with hyperthyroidism and are directly correlated between than and with indexes of thyroid function; therefore, they may be regarded as peripheral indexes of hyperthyroidism.


[Morphology and delta-endotoxin proteins of Bacillus thuringiensis from soils and their toxicities to insects]. 94 strains of Bacillus thuringiensis were isolated from soils in southwest and northwest of China. The morphology of cells, spores and parasporal crystals of these strains was investigated under transmission and scanning electro-microscope. Proteins of delta-endotoxins from all strains were analysed by rapid SDS-PAGE. 9 species of insects in Lepidoptera, Coleoptera and Diptera were tested for assay of delta-endotoxins. Some kinds of parasporal crystals were quite different in form and in composition of protein from those reported before. Most of strains were nontoxic to all of 9 species used in bio-assay. Some strains were very effective in species of Coleoptera or Noctuidae.


[Isolated ACTH deficiency and transient GH deficiency. Presentation of a case]. In a 46 year old man, who arrived at our observation suffering for three months from considerable increasing weakness and progressive impairment of libido, we documented a condition of secondary hypocorticism due to an isolated ACTH deficiency associated with a reduced somatotropin reserve, the last improved after treatment with corticosteroids. We found low serum levels of ACTH and cortisol, good response of adrenal glands to corticotropin depot, normalization of the clinical board during glucocorticoid replacement. Stimulating test with CRH (corticotropin releasing hormone) did not cause a response in ACTH, suggesting the presence of primitive damage of the hypophyseal corticotroph cells.


[Plasmid localization and cloning of restriction modification genes from Citrobacter freundii 4111 strain]. Over 60 producing strains of restriction endonucleases type II have been found among 500 different strains, mostly Enterobacteriaceae. The strain Citrobacter freundii 4111 produces restriction endonuclease CfrBI, a new isoschisomer of StyI. The genes of the restriction-modification system CfrBI were located on the multicopy plasmid pZE8 containing the Co1E1-type replicon and cloned to E. coli K802. The deletion variant of 3.2-kb pZE8 which contains intact restriction-modification and a DNA fragment responsible for autonomous plasmid replication was selected among the recombinant plasmids. The strain with higher R. CfrBI production (at least 10,000,000 U/g cells, which is 500-fold higher than the wild strain) was constructed.


Evolution, immune responses and stress: studies on molluscan cells. Using a variety of techniques we found the presence of ACTH and opioid-like molecules in the phagocytic spreading hemocytes (SH) and the serum of the mollusc Planorbarius corneus. In vitro experiments have shown that ACTH and beta-endorphin exert chemotactic activity on SH, while ACTH, but not beta-endorphin increases the phagocytic activity of SH. Moreover, ACTH and CRF provoked the release of biogenic amines from the hemocytes, mimicking a proto-stress type of response. Thus, the same mobile cell is capable of immune and proto-stress responses by using as mediators neuropeptides which remained fundamentally similar throughout evolution.


[Use of non-radioactive biotin-labelled probes for detecting hepatitis A virus]. The biotin-labeled DNA probes were constructed on the basis of the hybrid bacteriophage M13nip 9 single-stranded DNA containing the fragments of the hepatitis A viral cDNA. The probes were biotin treated by chemical modification of the DNA by the peraminating reagent or photochemically. The labeled DNA probes were used in molecular hybridization experiments with the nuclear acids fixed on the nitrocellulose filters. The biotin treated DNA was determined by the avidin-gold colloid conjugate with the subsequent physical silver amplification or by the streptavidin-alkaline phosphatase conjugate. The sensitivity of both probes was identical and permitted the determination of 5 x 10(-11)-5 x 10(-12) g of the control DNA and 10(-9) g of the hepatitis A virus. The developed test systems were used for detection of the viral RNA in blood from patients.


Bidirectional transmission in the cerebrobuccal connective of Aplysia during feeding. The neuronal activity of the cerebrobuccal connective (CBC) of Aplysia was recorded, using 2 implanted electrodes, under three conditions; 1) in the absence of feeding behaviour, 2) during appetitive feeding behaviour and 3) during consummatory feeding behaviour. Cross-correlation analysis of the recordings was then performed to subdivide spikes on the basis of their direction and speed of propagation. This revealed differences in the neuronal activity during the 3 conditions. There was little activity in the CBC when animals were not feeding. During appetitive and consummatory feeding behaviour the activity in the CBC increased. Units travelling in each direction were present, but with differential activity during the 2 behavioural patterns.


[Anti-CMV antibody affinity in renal transplant recipients]. Variations in anti-CMV antibody affinity have been studied in 106 renal transplant patients. Maturation of immune response has been followed during two years after transplantation evaluating antibody affinity by ELISA before and after urea denaturation treatment. In primary infections while the antibody titer rises, the resistance to denaturing treatment rises as well, indicating an increased antibody affinity. In patients already seropositive at transplantation, the increase of antibody affinity has also been found: comparing the affinity index (AI) at transplantation and one year later, only 16.5% of patients showed an AI value between 80 and 100 at transplantation, whereas after one year 62.3% of patients reached such AI values (p = 0.0001).


Regulation of transmitter release by presynaptic receptors at a cholinergic neuro-neuronal synapse. The modulation of evoked transmitter release by presynaptic receptors was studied at an identified cholinergic synapse in the buccal ganglion of Aplysia. Two auto-receptors affecting acetylcholine release in opposite ways were identified. Additionally acetylcholine (ACh) release was found to be facilitated by the peptide FLRFamide and inhibited by histamine. Ca2+ channels appeared as the final effectors controlled by these non-cholinergic presynaptic receptors whereas the activation of cholinergic presynaptic receptors did not affect the Ca2+ influx. The intracellular pathway activated by FLRFamide receptors was investigated in detail. The facilitation of transmitter release induced by this peptide was prevented by bath application of H-7, a protein kinase C inhibitor. Moreover, a diacylglycerol analog mimicked the action of FLRFamide. These results suggest that activation of protein kinase C leading to the phosphorylation of Ca2+ channels could be the mechanism through which presynaptic FLRFamide receptors increase evoked quantal release of acetylcholine at this synapse.


Pharmacological profiles of the GABA and acetylcholine receptors from the nematode, Ascaris suum. The pharmacological profiles of the acetylcholine and GABA receptors of Ascaris muscle have been investigated. Acetylcholine excites the muscle through activation of a nicotinic receptor which resembles the mammalian ganglionic receptor. DMPP is a potent agonist and alpha-bungarotoxin is a weak antagonist. GABA inhibits the muscle through activation of a chloride-linked receptor which in terms of agonist profile resembles the mammalian GABA-A receptor. However, bicuculline is inactive and picrotoxin is a very weak antagonist. Avermectin acts as a non-competitive antagonist at this GABA receptor with an IC-50 value in the low microM range.


Low concentrations of neuroactive peptides modulate cholinergic transmission and cyclic AMP levels in Helix aspersa. 1. FMRFamide and the Catch relaxing peptide (CARP) at 0.01-0.1 nM modulate acetylcholine (ACh) induced currents of identified neurons of Helix aspersa, but have no direct effect on membrane current and conductivity. 2. Both FMRFamide and CARP noncompetitively inhibit ACh Cl- responses while having either no effect or increasing ACh Na+/K+ response. 3. The inhibitory effect of FMRFamide and CARP on the ACh Cl- response was eliminated following pretreatment with forskolin (20 microM), an activator of adenylate cyclase. 4. Ascaris peptide (ASC) and a synthetic CARP analogue NORL-CARP, in which the amino acid methionine is replaced by either leucine or norleucine, at 1-10 microM, showed no effect on responses to ACh. 5. FMRFamide and CARP, at 10 nM, increased cAMP levels to 240% and 148% respectively above resting basal cAMP levels, while ASC and NORL-CARP had no significant effective. 6. Our results suggest that FMRFamide and CARP, in low concentrations, modulate ACh responses of Helix neurons, possibly through changes in cAMP levels. They also indicate the importance of the presence of methionine in these neuroactive peptides.


Role of protein kinase system in the signal transduction of stretch-mediated myocyte growth. To examine the molecular mechanisms by which mechanical stimuli induce protooncogene expression, we cultured rat neonatal cardiocytes in deformable dishes and imposed an in vitro mechanical load by stretching the adherent cells. Myocyte stretching increased total cell RNA content and mRNA levels of c-fos and skeletal alpha-actin followed by activation of protein synthesis. CAT assay indicated that sequences containing a serum response element were required for efficient transcription of c-fos gene by stretching. This accumulation of c-fos mRNA was suppressed by protein kinase C inhibitors at the transcriptional level and was inhibited markedly by down-regulation of protein kinase C. Moreover, myocyte stretching increased inositol phosphate levels. These findings suggest that mechanical stimuli might directly induce protooncogene expression, possibly, via protein kinase C activation. Furthermore, we observed the activation of mitogen activated protein (MAP) kinase by myocyte stretching. This result suggest that MAP kinase activation might increase the efficiency of protein synthesis in ribosomes induced by mechanical stimuli.


Diastolic dysfunction in pressure-overload hypertrophy and its modification by angiotensin II: current concepts. Cardiac hypertrophy is an adaptive response to an increased load imposed on the myocyte which allows the heart to perform increased work while maintaining normal myocardial fiber stress and shortening in systole. A deleterious consequence of pressure-overload hypertrophy is the prolongation of Ca(2+)-sensitive force inactivation (impaired myocardial relaxation) which is related to intrinsic alterations in cytosolic Ca2+ transport and reuptake in diastole. Additional factors appear to adversely modify myocardial relaxation in the hypertrophied heart, including the imposition of ischemia. There is also evidence that the expression and activity of the cardiac tissue renin angiotensin system (RAS) may be modified in the hypertrophied heart and contribute to diastolic dysfunction. Recent studies have demonstrated the presence of increased cardiac angiotensin converting enzyme (ACE) mRNA expression and activity in animal models of hypertrophy, including the aortic-banded rat with compensatory pressure-overload hypertrophy and rats with post-infarction remodeling. In the beating, isovolumic aortic-banded rat heart, the increased intracardiac activation of angiotensin I to II has been shown to be associated with a dose-dependent depression of diastolic relaxation. Preliminary studies suggest that the depression of diastolic function by angiotensin II in the hypertrophied heart can be prevented by the specific inhibition of cardiac ACE. In addition, the well-recognized susceptibility of the hypertrophied heart to severe ischemic diastolic dysfunction also appears to be favorably modified by the inhibition of cardiac ACE activity. The mechanisms responsible for the adverse effects of angiotensin II on diastolic relaxation in the hypertrophied heart are likely to be complex.(ABSTRACT TRUNCATED AT 250 WORDS)


[Cancer of the uterine cervix in Togo. Epidemiological observations and anatomopathological characteristics]. Fifty consecutive cases of cervical cancer from the Afagnan Hospital in Togo have been reviewed in their clinical features, and in ten out of these and five additional ones from the year 1979 the histological specimens were reexamined. The high rate of late diagnosis was the most striking clinical data observed, while the histopathological study revealed a high quota of undifferentiated or rare patterns, very different from the types commonly observed in Europe. In spite of the very poor public health level of the population, an almost complete absence of histological signs of HPV infection in the neoplastic tissue should be remarked.


Sympathetic modulation of the cardiac myocyte phenotype: studies with a cell-culture model of myocardial hypertrophy. Myocardial hypertrophy is the common endpoint of many cardiovascular stimuli such as hypertension, myocardial infarction, valvular disease, and congestive failure. Catecholamines have long been implicated in the pathogenesis of myocardial hypertrophy, however, it is very difficult to sort out catecholamine mechanisms in vivo. We have developed a cell-culture model which excludes hemodynamic effects and allows the assignment of receptor specificity to catecholamine effects. Utilizing this system, we have shown that stimulation of the alpha 1 adrenergic receptor leads to the development of myocardial hypertrophy and results in the selective up-regulation of the fetal/neonatal mRNAs encoding skeletal alpha-actin and beta-MHC, a pattern similar to that seen with hypertrophy in-vivo. Utilizing a co-transfection assay, we have also obtained data that suggest that the beta-PKC isozyme is in a pathway regulating transcription of the beta-MHC isogene. Beta adrenergic stimulation of the cultured cardiac myocytes also results in a modest degree of hypertrophy, however, this effect may be dependent upon myocyte contractile activity and may involve, at least in part, the non-muscle cells present in the culture system.


Modulation of myocardial sarcoplasmic reticulum Ca(++)-ATPase in cardiac hypertrophy by angiotensin converting enzyme? Myocardial hypertrophy in response to hemodynamic overload is an established risk factor for cardiovascular morbidity and mortality. Partially, this may be due to alterations in cardiac gene expression, resulting in a more fetal-like myocyte phenotype with a fragile Ca(++)-homeostasis. Depressed expression of the sarcoplasmic reticulum Ca(++)-ATPase is the hallmark of this overload phenotype, contributing to prolonged cytosolic Ca(++)-transients, disturbed diastolic relaxation, altered force-frequency relation, and probably, electrophysiologic instability with susceptibility to malignant arrhythmias. Since angiotensin II is a growth-promoting factor in several cellular systems, the local formation of angiotensin II within the myocardium might contribute to the trophic response and the phenotype shift of overloaded myocardium. Several observations are consistent with this hypothesis: the cardiac expression of ACE and angiotensinogen is enhanced in experimental myocardial overload and in human endstage congestive heart failure; prolonged observations of experimental cardiac overload with hypertrophy-induced putative normalisation of myocardial systolic wall stress demonstrated a renormalization of ventricular tissue ACE activity and of ventricular sarcoplasmic Ca(++)-ATPase expression and activity; normalizing ventricular tissue ACE activity in experimental cardiac overload by chronic nonhypotensive ACE inhibitor therapy caused a parallel partial normalization of hypertrophy and underexpression of sarcoplasmic CA(++)-ATPase. This partial normalization of myocyte Ca(++)-homeostasis in overload hypertrophy by non-hypotensive chronic ACE-inhibition is attenuated by concomitant chronic application of bradykinin-2 receptor blockade, indicating an involvement of altered bradykinin metabolism in the phenotype modulation due to chronic ACE inhibition. While these observations are consistent with a direct influence of local ACE activity on the sarcoplasmic reticulum, the cell type contributing to the enhanced ACE expression in overload and the specific mechanism of this influence are unknown.


The regulation of calcium cycling in stressed hearts. Myothermal measurements of tension-independent heat are used to calculate the quantity of calcium released during isometric contraction and the rate at which it is removed in control, thyrotoxic and pressure-overloaded rabbit hearts. Experiments were carried out at 30 degrees C. In control rabbit hearts 41.0 +/- 7.0 nmoles/g Ca++ was released into the cytosol for each beat, while the rate at which the Ca++ was removed from the cytosol was 24.4 +/- 4.4 nmoles/g sec. In the presence-overloaded preparations, the amount of calcium released and the rate of calcium removal were 41% and 40% of control values. This reduction was correlated with the mRNA levels for the sarcoplasmic reticulum (SR) Ca++ ATPase, phospholamban and the ryanodine receptor. The depression was also correlated with a reduction in SR Ca++ ATPase protein expression. In thyrotoxic hearts compared with controls, with each activation there is an increase in the amount of calcium liberated into the cytosol (39%) and the rate of calcium removal (31%). This increase is correlated with an increase in the mRNA and protein expression for the SR Ca++ ATPase as well as the mRNA for the ryanodine receptor. Calsequestrin mRNA was unchanged in all of the experimental preparations. It is suggested that the alteration in the calcium cycling proteins offers at least a partial explanation for the changes in calcium cycling measured in response to the stresses applied.(ABSTRACT TRUNCATED AT 250 WORDS)


A processing strategy for automated Papanicolaou smear screening. A multilayer processing strategy was developed for the automatic screening of conventionally prepared Papanicolaou smears. The processing stages include image segmentation, feature extraction, object classification and slide classification. Mathematical morphology functions were implemented in hardware with custom-built gate array processors for image segmentation. There were 68 features used for classifier training. In object classification we combined the evidential supports of a binary decision tree classifier and a multilayer perceptron classifier to achieve an integrated decision. In this feasibility study, 449 conventionally prepared cervical Papanicolaou smears were tested in a prototype research system between January and May 1991. The 95% confidence interval for the slide false-negative rate was 1-9%, and the 95% confidence interval for the slide sort rate was 45-55%. The estimated sort rate for clearly normal slides is within the range required for a cost-efficient screening system, and the estimated false-negative rate for premalignant and malignant smears is an improvement over published false-negative rates for human performance. Several performance improvement efforts are still under way. We expect that they will result in a vastly reduced slide false-negative rate.


Cross-activation: overriding cAMP/cGMP selectivities of protein kinases in tissues. cAMP- and cGMP-dependent protein kinases are homologous proteins and are predicted to exhibit very similar three-dimensional structures. Their cyclic nucleotide binding domains share a high degree of amino acid sequence identity. cAMP- and cGMP-dependent protein kinases are activated relatively specifically by cAMP and cGMP, respectively; and a single alanine-threonine difference between cAMP- and cGMP-binding domains partially accounts for this specificity. Thus, it would be expected that cAMP and cGMP mediate separate physiological effects. However, owing in part to the lack of absolute specificity of either enzyme and to the relatively high level of cAMP or cGMP in certain tissues, it is also possible that either cyclic nucleotide could cross-activate the other kinase. Increases in either cAMP or cGMP cause pig coronary artery relaxation. However, only cGMP-dependent protein kinase specific cyclic nucleotide analogues are very effective in causing relaxation, and cAMP elevation in arteries treated with isoproterenol or forskolin activates cGMP-dependent protein kinase, in addition to cAMP-dependent protein kinase. Conversely, increases in either cAMP or cGMP cause Cl- secretion in T-84 colon carcinoma cells, and the cGMP level in T-84 cells can be elevated sufficiently by bacterial enterotoxin to activate cAMP-dependent protein kinase. These results imply specific regulation of cAMP- and cGMP-dependent protein kinases by the respective cyclic nucleotides, but either cyclic nucleotide is able to cross-activate the other kinase in certain tissues.


Insulin modulation of human apolipoprotein B mRNA translation: studies in an in vitro cell-free system from HepG2 cells. Insulin modulation of apolipoprotein B gene expression was studied at the translational level by the use of a cell-free translation system from a hepatoma cell-line, HepG2. Extracts of HepG2 cells lysed with lysolecithin were found to have high in vitro protein synthesizing activity utilizing endogenous mRNA. The level of peptide chain initiation was high, as suggested by a significant inhibition of translation by edeine. The translation products of endogenous mRNA in HepG2 cell-free lysate were probed with anti-apolipoprotein B antibodies to investigate its synthesis. A 550 kilodalton (kDa) polypeptide was selected by a polyclonal antibody, as well as a monoclonal antibody, against the C-terminal end of apolipoprotein B molecule. This in vitro synthesized polypeptide was also found to compare well in size with the in vivo product. The HepG2 lysate was also shown to efficiently synthesize in vitro a number of other proteins including albumin, apolipoprotein E, apolipoprotein A1, and actin. The in vitro synthesis of polypeptides as large as 500 kDa was unexpected and has not previously been demonstrated in a cell-free system. The HepG2 translation system was used to investigate the effect of insulin on the in vitro translation of apolipoprotein B. Lysates prepared from HepG2 cells treated with insulin were found to have lower translational activity (by an average of 52.3%) for apolipoprotein B compared with lysates from control untreated cells. In vitro synthesis of actin and apolipoprotein E were unaffected under these conditions. The insulin-stimulated decline in in vitro apolipoprotein B synthesis was not due to a change in apolipoprotein B mRNA levels as determined by slot- and Northern-blot analyses, suggesting that the inhibitory effect of insulin may be exerted partly at the level of apolipoprotein B mRNA translation.


Apolipoprotein synthesis and secretion in Hep G2 cells: effects of monensin and cycloheximide. Hep G2 cells were used to study the relationship between apolipoprotein synthesis and secretion, as revealed by their interaction with agents modulating these processes. Cycloheximide inhibited the secretion of both apolipoproteins (apo) AI and B, but the reduction in apo AI secretion was evident at earlier times. Monensin also inhibited secretion of apo AI and apo B, but only apo AI accumulated intracellularly. Pulse-chase studies showed that, at concentrations of monensin that had no effect on total protein synthesis, apo B synthesis was specifically inhibited. Triacylglycerol synthesis was inhibited to the same extent as apo B synthesis, but this preceded the latter inhibition and unlike apo B there was an accumulation of intracellular triglyceride. These results suggest that distinctive mechanisms modulate the synthesis and secretion of apo AI and apo B, and that apo B synthesis can be specifically inhibited by mechanisms that initially block triglyceride production.


Dephosphorylation of the retinoblastoma protein during differentiation of HL60 cells. Immunoprecipitated retinoblastoma protein from HL60 cells migrated as a series of bands during electrophoresis. The heterogeneity appeared to be generated by phosphorylation of the retinoblastoma protein. Treatment of the cells with the phorbol ester, tetradecanoyl phorbol acetate (TPA), resulted in both a loss of the heterogeneity of the pRB species and a significant decrease in the level of pRB phosphorylation. These changes accompanied differentiation of the HL60 cells into macrophages. Treatment of the cells with dibutyryl cAMP also resulted in dephosphorylation of pRB as well as cell cycle arrest, although no recognizable differentiation occurred. These results are consistent with a model in which TPA and dibutyryl cAMP dependent pathways can activate pRB by altering its phosphorylation.


Tumour necrosis factor and cancer.

Tumour necrosis factor and cancer. TNF is a cytokine whose diverse actions are dependent on the local microenvironment. As a member of the cytokine network, TNF plays an important role in infection and inflammation, but excessive and deregulated production can contribute to disease processes. Likewise in malignant disease, TNF may have a role in cancer therapy and contribute to host response against tumours, but it may also be involved in the progression and spread of the cancer. In experimental models, recombinant TNF can induce significant haemorrhagic necrosis, localised to the tumour vasculature and specific tumour immunity. Although the historical background and preclinical data are promising, systemic therapy with TNF in human cancer has proved highly toxic and is inactive against all tumour types so far tested. Local therapy, particularly isolated limb perfusion, has resulted in complete and long lasting tumour regressions with necrotic activity confined solely to the tumour vascular bed. However, in several animal models, TNF contributes to malignant progression and there is evidence that TNF may have autocrine or paracrine actions in human ovarian cancer.


Ciliary neurotrophic factor and its receptor complex. Ciliary neurotrophic factor (CNTF), originally identified for its ability to promote survival of neurons of the ciliary ganglion, is now known to have additional survival and differentiative actions on cells of the nervous system. CNTF is, however, unrelated in structure to the nerve growth factor family of neurotrophic factors. Instead, CNTF is distantly related to, and in fact shares receptor components with, a number of hemopoietic cytokines. This review focuses on the biological actions of CNTF, the shared and unique features of the CNTF receptor complex and signaling pathways, and the distribution of CNTF and its receptor during development, in the adult and in response to injury.


Oncostatin M.

Oncostatin M. Oncostatin M (OSM) was initially identified as a polypeptide cytokine which inhibited the in vitro growth of cells from melanoma and other solid tumors. OSM shows significant similarities in primary amino acid sequence and predicted secondary structure to leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), granulocyte colony-stimulating factor (G-CSF), interleukin 6 (IL-6), and interleukin 11 (IL-11). Analysis of the genes encoding these proteins reveals a shared exon organization suggesting evolutionary descent from a common ancestral gene. Recent data indicates that OSM also shares a number of in vitro activities with other members of this cytokine family. The overlapping biological effects appear to be explained by the sharing of receptor subunits.


Regulation of phospholipase C isozymes.

Regulation of phospholipase C isozymes. Phosphatidylinositol bisphosphate hydrolysis is an immediate response to many hormones, including growth factors. The hydrolysis of phosphatidylinositol bisphosphate is catalyzed by phosphatidylinositol-specific phospholipase C. A number of phospholipase C isozymes have been identified. Different isozymes are activated by different receptor classes. This review will summarize the different isozymes of phospholipase C, and the current knowledge of the mechanisms by which phospholipase C activity is modulated by growth factors.


Effectiveness of reciprocal-action instrumentation for polishing composite resin: an in vitro study. This in vitro study investigates the use of reciprocal-action instrumentation for polishing composite resin restorative materials. Electron photomicrographs were made of surfaces of a microfilled and a hybrid composite resin restorative material polished by various reciprocal-action polishing procedures, including several types of polishing pastes used with nonabrasive plastic tips. The best polish on the microfilled composite resin restorative material tested was produced by ending with 3-mu diamond polishing paste. The best polish on the hybrid composite resin restorative material tested was produced by ending with 0.5-mu diamond polishing paste. The surfaces produced were comparable to those produced by polishing discs.


The relationship of gap junctions and compaction in the preimplantation mouse embryo. In the mouse embryo, gap junctions first appear at the 8-cell stage as compaction is about to take place. Compaction of the embryo is important for the differentiation of the first two cell types; the inner cell mass and the trophectoderm. Our studies examine the contribution of gap junctional communication at this stage of development. We have characterised the normal sequence of appearance of gap junction protein and its distribution. The extent of communication as shown by the passage of dye between cells has been recorded in both normal embryos and embryos treated with drugs that influence gap junctional communication. Comparisons have been made with embryos that express a lethal gap junction defect and attempts were made to rescue such embryos by increasing their gap junction communication.


Camptothecin hyper-resistant P388 cells: drug-dependent reduction in topoisomerase I content. A subline of P388 leukemia made 10-fold resistant to camptothecin (CPT) by serial passage in drug-treated mice was adapted to growth in tissue culture and made hyper-resistant to CPT by passage in the presence of increasing concentrations of the drug. Cells were obtained that were 1,000-fold resistant to CPT, compared to wild-type P388 cells. Neither topoisomerase I mRNA nor 100 kDa topoisomerase I enzyme was detectable in these cells, and topoisomerase I activity extracted from nuclei was less than 4% of that extracted from nuclei of wild-type cells. An immunoreactive 130 kDa protein that could be an altered, inactive form of topoisomerase I was evident in the hyper-resistant cells. In addition, the cells deficient in topoisomerase I contained enhanced topoisomerase II activity. Maintenance of the hyper-resistant phenotype required continued exposure to CPT; growth in its absence led to loss of hyper-resistance, increased topoisomerase I content and activity, and decreased topoisomerase II activity. The sensitivity of the cells to killing by a number of inhibitors of topoisomerases I and II was consistent with these observations. Thus, P388 cells have the potential to become highly resistant to CPT by severely curtailing topoisomerase I expression; in these circumstances, topoisomerase I and II activities are regulated coordinately.


Uptake of exogenous human papilloma virus L1 DNA by oocytes and detection by the polymerase chain reaction. The purpose of this study was to determine if oocytes were capable of taking up exogenous DNA such as human papillomaviral (HPV) DNA and evaluate the zona pellucida as a barrier to the entry of foreign DNA into the oocyte. The experiment consisted of four groups of hamster oocytes exposed to HPV DNA fragments: Group A, zona-free oocytes (n = 5); Group B, oocytes with an intact zona pellucida (n = 5); Group C, oocytes fixed in 4% buffered formalin solution for 20 min (n = 5); and Group D, zona-free oocytes (n = 4). Group C oocytes served as an internal control to ensure adequate washing of the oocytes after incubation. The zona pellucida was not a barrier to foreign DNA molecules. The PCR did not detect L1-HPV and beta-globin gene sequences in the untreated hamster oocyte. Uptake of the smaller DNA fragments such as that amplified from the beta-globin region was independent of active oocyte cell processes. Oocytes cultured in vitro can passively take up exogenous DNA fragments. The results suggest a possible role of oocytes as vectors for foreign DNA.