Protein kinase-ζ interacts with munc18c: role in GLUT4 trafficking

Aims/hypothesis Insulin-stimulated glucose transport requires a signalling cascade through kinases protein kinase (PK) C/ and PKB that leads to movement of GLUT4 vesicles to the plasma membrane. The aim of this study was to identify missing links between the upstream insulin-regulated kinases and the GLUT4 vesicle trafficking system.Materials and methods A yeast two-hybrid screen was conducted, using as bait full-length mouse munc18c, a protein known to be part of the GLUT4 vesicle trafficking machinery.Results The yeast two-hybrid screen identified PKC as a novel interactor with munc18c. Glutathione S transferase (GST) pull-downs with GST-tagged munc18c constructs confirmed the interaction, mapped a key region of munc18c that binds PKC to residues 295–338 and showed that the N-terminal region of PKC was required for the interaction. Endogenous munc18c was shown to associate with endogenous PKC in vivo in various cell types. Importantly, insulin stimulation increased the association by approximately three-fold. Moreover, disruption of PKC binding to munc18c by deletion of residues 295–338 of munc18c or deletion of the N-terminal region of PKC markedly inhibited the ability of insulin to stimulate glucose uptake or GLUT4 translocation.Conclusions/interpretation We have identified a physiological interaction between munc18c and PKC that is insulin-regulated. This establishes a link between a kinase (PKC) involved in the insulin signalling cascade and a known component of the GLUT4 vesicle trafficking pathway (munc18c). The results indicate that PKC regulates munc18c and suggest a model whereby insulin triggers the docking of PKC to munc18c, resulting in enhanced GLUT4 translocation to the plasma membrane.     

Protein Kinase C Modulation of Cyclic GMP in Rat Neonatal Pulmonary Vascular Smooth Muscle

Protein kinase C (PKC) has been implicated in the control of vascular tone and mitogenesis in the adult pulmonary vasculature, but little is known about the role of PKC in the neonatal pulmonary vasculature. In addition, the vasodilator nitric oxide (NO) is important in the transition of the pulmonary circulation from fetal to postnatal life, and it is thought that attenuated production of NO and therefore, cGMP may contribute to the pathophysiology of a variety of forms of neonatal pulmonary vascular disease states. Although evidence exists for an interaction between PKC and NO in the adult pulmonary vasculature, the identification of specific roles for PKC in neonatal pulmonary vascular smooth muscle (NPVSM) has not been determined, and no studies have been done on the modulation of cGMP by PKC in NPVSM. Accordingly, immunoblot analysis revealed the expression of the , , , and PKC isozymes in NPVSM. Treatment of NPVSM with 10nM 4- phorbol myristate acetate (PMA), a PKC activator, induced translocation of PKC, and PKC from the soluble to the particulate fraction, while exposure to 10nM endothelin-1 (ET-1), a potent vasoconstrictor and mitogenic substance, caused translocation of PKC and PKC from the soluble to the particulate fraction. Sodium nitroprusside (SNP) significantly increased intracellular cGMP levels, an effect attenuated by PMA but not by ET-1. In addition, pretreatment with the specific PKC isozyme antagonist Go 6983 blocked the effect of PMA on cGMP levels. Collectively, these data demonstrate the expression and activation of multiple PKC isozymes in NPVSM, and indicate that PKC inhibits SNP-stimulated cGMP production in NPVSM. These data also suggest that complex intracellular signaling pathways by specific PKC isozymes may be important in the development of neonatal pulmonary vascular function.     

Interleukin-1β Inhibits Growth Factor-Stimulated Restoration of Wounded Rat Gastric Epithelial Cell Monolayers

We examined the effect of interleukin-1(IL-1) on spontaneous and enhanced restoration(cell migration and proliferation) using an in vitrowound model comprising a confluent monolayer of ratgastric epithelial RGM1 cells. Repair of an artificialwound in a cell monolayer was found to be time- andconcentration-dependent when the cells were incubatedwith epidermal growth factor (EGF) or transforming growth factor (TGF)- alone for up to 24hr. The growth factors also stimulated DNA synthesissignificantly for 24 hr in a concentration-relatedmanner. IL-1 had no effect on wound restoration in the absence of the growth factors. However, itmarkedly inhibited the restoration enhanced by EGF andTGF-, the inhibition being about 60% and 70%,respectively. In addition, IL-1 significantly reduced the DNA synthesis stimulated by thegrowth factors. The EGF- and TGF--enhancedrestoration was reduced by about 30% by mitomycin C,which potently inhibited the stimulated DNA synthesis.Mitomycin C had no effect on the spontaneous restoration.Even when treated with mitomycin C, the inhibitoryeffect of IL-1 on the enhanced wound repair wasstill observed; however, the extent of the inhibition was decreased. These results indicate thatIL-1 inhibits the migration as well as theproliferation of gastric epithelial cells enhanced byEGF and TGF-, resulting in a failure of woundhealing.     

Role of Central Interleukin-1β in Gastrointestinal Motor Disturbances Induced by Lipopolysaccharide in Sheep

Cytokines are involved in the symptoms of theacute phase response induced by infectious diseases inhumans as well as in animals, and interleukin-1(IL-1 ) has a pivotal role in these changes. The role of central IL-1 in the gastrointestinalhypomotility and fever evoked by intravenousadministration of lipopolysaccharide (LPS) and themechanisms involved, were investigated in sheep as anexperimental model. LPS (0.1 g/kg, intravenously)induced gastrointestinal hypomotility and fever thatwere significantly reduced by priorintracerebroventricular administration of IL-1receptor antagonist protein (IL-1ra, 2 g/kg). The effects of LPS were mimickedby intracerebroventricular IL-1 (50 ng/kg),whereas IL-1 injected intravenously at the samedose only caused a slight and transient fever withoutmodifying the gastrointestinal motility. Priorintracerebroventricular administration of thecyclooxygenase inhibitor indomethacin (100 g/kg) butnot the corticotropin-releasing factor (CRF) receptorantagonist -helical CRF_9-41 (5 g/kg) blocked alleffects evoked by both LPS and IL-1. These resultssuggest that in sheep, LPS induces digestive motordisturbances through a central release of IL-1 andprostaglandins.     

Prinzipien der TNM-Klassifizierung

Zusammenfassung Grundprinzip des TNM-Systems ist Deskription statt Interpretation. Begriffe wie Früh- und Spätstadium sind radikal zu streichen. Bezugsbasis für die interdisziplinäre Vergleichsplanung therapeutischer Ergebnisse ist der prätherapeutisch erhobene Befund. Ausnahmen bestätigen diese Regel, nämlich dort, wo es sich um nicht zugängliche Tumoren handelt. Hier hat die chirurgisch-pathologische Befunderhebung bzw. Klassifizierung grundlegende und nicht mehr wie ursprünglich ergänzende Bedeutung, wie am Beispiel Magenkarzinom ersichtlich.Was die Symbole TNM im einzelnen bedeuten, wird beschrieben. Ebenso wird kurz auf die Bedeutung des Grading (G) und Staging eingegangen.

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Principles of TNM classification

Summary The basic principle of the TNM system is to describe and not to interpret. Terms such as early case and late case should be avoided. The basis for the interdisciplinary comparison of therapeutic results are the pretherapeutic findings. Exceptions to that rule are those cases in which the origin of tumours remains unknown. Then the surgico-pathological diagnoses and classifications are decisive and lose their original ancillary character, as in carcinoma of the stomach.The significance of the symbols TNM is described. The importance of Grading (G) and Staging is mentioned.

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Referat gehalten auf der 11. wissenschaftlichen Tagung der Deutschen Krebsgesellschaft Hannover 1971.     

Comparative and combined effects of transforming growth factors α and β, interleukin-1 and interferon-γ on rheumatoid synovial cell proliferation,glycolysis and prostaglandin E production

Summary Enhanced cell proliferation, glycolysis and prostaglandin E production are all characteristic features of rheumatoid synovial tissue. The interrelationships of these three cellular parameters have been examined using rheumatoid synovial fibroblasts and their responses to specific cytokines in vitro. Transforming growth factor (TGF) caused a more than threefold increase in synovial cell proliferation whilst transforming growth factor (TGF), interleukin-1 (IL-1) and interferon- (IFN-) produced only marginal changes. The combined addition of IL-1 with TGF resulted in an enhanced proliferative response comparable with that produced by TGF. Glycolysis, estimated by glucose utilisation and measurements of the glycolytic regulatory metabolite fructose 2,6-bisphosphate was significantly stimulated by TGF, IL-1 and IFN-, but less so by TGF. Prostaglandin E production was significantly increased by IL-1 to an extent much greater than that produced by TGF or TGF, although the combined addition of IL-1 with either TGF or resulted in a synergistic increase in PGE production, a response partly diminished by the addition of IFN-. These findings suggest that the extent to which a cytokine stimulates glycolysis is not consistently related to its mitogenicity, and that cytokine combinations which stimulate high levels of PGE production (a growth inhibitor) will not necessarily be associated with a reduced rate of cellular proliferation in cultured, adherent, rheumatoid synovial fibroblasts.     

B cell adrenoceptors and sulphonylurea-induced insulin release in mouse islets

Summary Interactions of tolbutamide and glibenclamide with B cell adrenoceptors have been reported. This study evaluated the possible role of such interactions in the stimulation of insulin release. Mouse islets were incubated in the presence of 10 mmol/l glucose alone or with tolbutamide (10 mol/l) or glibenclamide (0.02 mol/l). At 0.01–10 mol/l, blockers of ₂-adrenoceptors (yohimbine, idazoxan) or ₁-adrenoceptors (prazosin) had practically no effect on glucose-induced insulin release and did not affect its potentiation by sulphonylureas, except for a slight increase by 10 mol/l prazosin and idazoxan. Nonspecific -blockers (phentolamine, dihydroergotamine) increased control release at 10 mol/l, but only the latter amplified the response to tolbutamide. Blockers of -adrenoceptors were tested at 0.1–100 mol/l: propranolol (₁, ₂), metoprolol (₁) and compound ICI 118-551 (₂). They increased glucose-induced insulin release at 100 mol/l but variably altered the effect of sulphonylureas. Blockers of adrenoceptors have, thus, no effect on insulin release in vitro at therapeutic concentrations. At high concentrations, they non-specifically affect the action of sulphonylureas. We conclude that an interaction with B cell adrenoceptors is not involved in the insulinotropic action of sulphonylureas.     

Selective A2A adenosine agonist ATL-146e attenuates acute lethal liver injury in mice

Background d-Galactosamine (GalN)/lipopolysaccharide (LPS)-induced liver injury is an experimental model of fulminant hepatic failure in which tumor necrosis factor- (TNF-) plays a pivotal role. We examined the effects of a highly selective adenosine A_2A receptor agonist (ATL-146e) on GalN/LPS-induced fulminant hepatic failure.Methods Mice were given an intraperitoneal dose of GalN (800mg/g body weight)/LPS (100ng/g body weight) with and without ATL-146e (0.01µg/kg) treatment. Liver injury was assessed biochemically and histologically. Also, TNF- levels in the serum were determined.Results The serum liver enzyme (ALT) level in vehicle-treated mice was 20960 ± 2800IU/ml and was reduced by 63% to 7800 ± 1670IU/ml by treatment with 0.01µg/kg per minute ATL146e, P < 0.05. Treatment with ATL-146e significantly reduced serum TNF- and greatly reduced inflammation assessed by histopathologic examination compared with control mice treated with GalN/LPS. ATL-146e also reduced lethality at 12h from 65% to 13%.Conclusion The present findings suggest that the highly selective adenosine A_2A receptor agonist (ATL-146e) prevents endotoxin-induced lethal liver injury by suppression of TNF- secretion.     

A Three-dimensional Analysis of Blood Vessels in Bronchioloalveolar Carcinoma

The three-dimensional architecture of blood vessels within lung adenocarcinomas has not been well studied. In 19 cases with bronchioloalveolar carcinoma with central fibrosis, we three-dimensionally examined blood vessel architecture in 150 m thick sections stained with elastin staining and anti-CD34 antibody. We examined four regions: normal alveoli and three regions within the tumor including an area adjacent to the normal alveoli (external area), an area in which tumor cells were replacing epithelial cells (replacement area), and a central fibrotic area (fibrotic area). Elastin staining showed that elastic fibers formed the framework of the alveoli, and the alveolar structure shrank more strongly to the center of the tumor due to folding of alveolar walls invaded by adenocarcinoma cells. We also measured three vessel parameters in these four regions. The vessel diameters were 4.08±1.10 m, 3.95±1.02 m, 5.04±1.56 m, and 6.11±2.23 m, the circumferences of those vessels seen as complete circles were 43.11±12.78 m, 43.71±12.87 m, 95.21±39.32 m, and 126.77±54.65 m; the lengths between vessel bifurcations were 13.28±3.08 m, 13.47±4.58 m, 24.91±9.66 m, and 41.82±28.08 m in the normal alveoli, and the external, replacement, and fibrotic areas, respectively. Blood vessel architecture changed such that the vessels became larger and coarser towards the center of the tumor. Our three-dimensional analysis suggests continuous remodeling of alveolar capillaries rather than angiogenesis within bronchioloalveolar carcinoma.     

The ability of heat stress and metabolic preconditioning to protect primary rat cardiac myocytes

Primary rat cardiocytes were subjected to either thermal preconditioning for 30 min at 43°C or 20 min metabolic preconditioning (10 mM deoxyglucose, 20 mM lactate, pH 6.5). Eighteen hours later cells were analysed either for hsp 70i expression or subjected to a subsequent lethal heat stress or simulated ischaemia (10 mM deoxyglucose, 20 mM lactate, 0.75 mM sodium dithionite, 12 mM potassium chloride, pH 6.5) for 2 hours and assessed for survival by trypan blue exclusion.Hsp 70i was induced over 100 fold by thermal preconditioning and 30 fold by metabolic preconditioning (p<0.001, p<0.05), hsp 90 was induced 2.71 fold and 2.24 fold (p<0.001, p<0.001) by thermal and metabolic preconditioning respectively, while hsp 60 was not induced by either treatment. Preconditioned cultures had improved survival against subsequent lethal heat stress or simulated ischaemia: Thermal preconditioning reduced death from 69.22% to 52.46% upon subsequent lethal heat stress and from 49.13% to 36.66% upon subsequent lethal simulated ischaemia. Metabolic preconditioning reduced cell death from 51.29% to 33.8% against subsequent lethal heat stress, and from 69.09% to 55.61% upon subsequent lethal simulated ischaemia. A second marker of cell death, the release of lactate dehydrogenase activity into the culture media, was reduced to 65% and 60% of control values for thermally preconditioned cells subjected to lethal heat or lethal simulated ischaemia respectively. Metabolically preconditioned cells demonstrated lactate dehydrogenase activity of 59% and 51% that of control values, when subjected to lethal heat or lethal simulated ischaemia respectively.Abbreviations hsp heat stress protein - hsp 70i inducible 70 kDa heat stress protein - LDH lactate dehydrogenase - PBS phosphate buffered saline     

Analysis of lymphocyte subsets in patients with aplastic anemia before and during immunosuppressive therapy

Summary To define the contribution of T-lymphocyte subsets in the development of aplastic anemia (AA), T-cell subpopulations including T cells, T cells, and TCS1-positive T cells, were analyzed by cytophotometry in the peripheral blood (PB) and bone marrow (BM) of patients with AA before and after 6 weeks of therapy with anti-lymphocyte globulin (ALG), methylprednisolone, and cyclosporin A (CSA). In nine patients with AA a significant decrease of PB- and BM-derived T cells was observed after 6 weeks of therapy as compared with normal controls. At diagnosis, the CD4/CD8 ratio in PB and BM of the patients did not differ from the ratio in the control population; however, a reversed ratio (< 1) was present in PB as well as in BM after weeks of therapy. Interestingly, lymphocytes expressing the T-cell receptor (TCR) were significantly decreased both before (PB 1.2±0.1%; BM 0.8±0.1%) and after 6 weeks of therapy (PB 0.7±0.1%; BM 0.7±0.1%) as compared with healthy controls (PB 2.4±0.2%; BM 2.3±0.2%). However, the proportion of the -T-cell subpopulation expressing the TCS1 phenotype was markedly increased before (PB 42±3.5%; BM 31±3%) and especially after 42 days of therapy (PB 77±12%; BM 45±2%) as compared with that in normal subjects (PB 19±2%; BM 9.7±0.8%). At present, follow-up is under evaluation to correlate these findings with hematological response. The pathophysiological significance of the observed alterations within the T-cell subsets and especially the T-cell populations will require further functional analyses, in particular since TCS1-positive T cells exhibit autoimmunological capacity.Presented at the annual meeting of the German Society for Hematology and Oncology, 4–7 October 1992, Berlin     

Local and regional immunotherapy of cancer with interleukin 2

Conclusion The results obtained in preclinical systems as well as the first clinical trials suggest that local IL-2 immunotherapy may represent a novel approach to the treatment of some neoplasms. However, the experimental results should be confirmed and substantially extended before definitive conclusions can be drawn. We hope that the data and considerations discussed in this article will facilitate thoughts concerned with future clinical IL-2 trials and be instrumental in the optimization of IL-2 cancer immunotherapy.The Journal of Cancer Research and Clinical Oncology publishes in loose succession Editorials and Guest editorials on current and/or controversial problems in experimental and clinical oncology. These contributions represent exclusively the personal opinion of the author     

“Bound” insulin: in vivo and in vitro biologic activity

Summary Partially purified preparations of human serum bound insulin were injected intraperitoneally or intravenously into intact fed, fasted or fasted-refed rats, or incubatedin vitro with their isolated epididymal adipose tissue. The biologic activity of bound insulin on muscle and adipose tissue,in vivo andin vitro, was compared with the activity of crystalline insulin standards. Bound and crystalline insulin injected intraperitoneally into rats stimulated the incorporation of glucose-u-¹⁴C into the glycogen of muscle and into the glycogen and fat of adipose tissue. The activities of both bound and crystalline insulin were affected by the nutritional state of the animals. Intravenous administration of partially purified bound insulin or crystalline insulin stimulated the incorporation of glucose-u-¹⁴C into the muscle glycogen of intact fed, fasted or fasted-refed rats and into the fat of the adipose tissue of fed or fasted-refed rats. Bound insulin, but not crystalline insulin stimulated fat synthesis in the adipose tissue of fasted rats. Bound or crystalline insulin, at the concentration used, failed to stimulate glycogen synthesis in adipose tissue. Bound insulin stimulated the oxidation of glucose into CO₂ in isolated rat adipose tissue and the incorporation of glucose-u-¹⁴C into the glycogen and fat of the adipose tissue. The effect of bound or crystalline insulin on isolated adipose tissue was also affected by the nutritional state of the animal.

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Zusammenfassung Teilweise gereinigte Präparate von gebundenem Insulin aus menschlichem Serum wurden normalen gefütterten, hungernden oder nach Hungern wiederaufgefütterten Ratten intraperitoneal oder intravenös injiziert oderin vitro mit dem isolierten epididymalen Fettgewebe dieser Ratten inkubiert. Die biologische Aktivität des gebundenen Insulins am Muskel und Fettgewebe,in vivo undin vitro, wurde mit der Aktivität von Standardlösungen kristallinen Insulins verglichen. Wurde gebundenes und kristallines Insulin Ratten intraperitoneal injiziert, kam es zu einer vermehrten Einlagerung von Glukose-U-¹⁴C in Muskelglykogen und in das Glykogen und Fett des Fettgewebes. Sowohl die Aktivität des gebundenen als auch des kristallinen Insulins wurden durch den Ernährungszustand der Tiere beeinflußt. Die intravenöse Verabreichung des teilweise gereinigten gebundenen Insulins oder des Kristallinsulins führte zu einer verstärkten Einlagerung von Glukose-U-¹⁴C in das Muskelglykogen der gefütterten, hungernden oder der nach Hungern wiederaufgefütterten Ratten und auch in das Fett des Fettgewebes der gefütterten und nach Hungern wiederaufgefütterten Ratten. Nur das gebundene Insulin, nicht das kristalline Insulin steigerte die Fettsynthese des Fettgewebes der hungernden Ratten. Weder das gebundene noch das kristalline Insulin führten bei den verwendeten Konzentrationen zu einer Steigerung der Glykogensynthese des Fettgewebes. Das gebundene Insulin stimulierte die Oxydation von Glukose zu CO₂ am isolierten Fettgewebe der Ratte und die Einlagerung von Glukose-U-¹⁴C in das Glykogen und Fett des Fettgewebes. Die Wirkung des gebundenen und des kristallinen Insulins auf das isolierte Fettgewebe wurde ebenfalls durch den Ernährungszustand der Tiere beeinflußt.

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Résumé Cette étude porte sur l'injection intrapéritonéale ou intraveineuse de préparations partiellement purifiées d'insuline liée de sérum humain. Des rats nourris ad libitum, mis à jeun, ou renourris après une période de jeûne, ont servi pour ces injections, et leur tissu adipeux épididymaire a servi à l'étude de l'effet in vitro de ces préparations. L'activité biologique de cette insuline liée a été comparée avec celle d'une insuline cristalline standard. L'insuline liée tout comme l'insuline cristalline stimule l'incorporation de glucose-u-C¹⁴ dans le glycogène du muscle et dansle glycogène et la graisse du tissu adipeux, à la suite de leur injection intrapéritonéale. L'état de nutrition des animaux injectés modifie les activités et de l'insuline liée et de l'insuline cristalline. L'injection intraveineuse d'insuline liée partiellement purifiée, ou d'insuline cristalline, stimule l'incorporation de glucoseu-C¹⁴ dans le glycogène musculaire de rats nourris ad libitum, à jeun, ou renourris après un jeûne prolongé. Elle stimule également l'incorporation du glucose dans la graisse du tissu adipeux de rats nourris ou renourris après jeûne. L'insuline liée seulement, non l'insuline cristalline, stimule la lipogénèse du tissu adipeux de rats à jeun. Tant l'insuline cristalline que l'insuline liée est restée sans effet, pour les dosages utilisés, sur la synthèse du glycogène dans le tissu adipeux. L'insuline liée stimule l'oxydation du glucose par le tissu adipeux isolé ainsi que son incorporation dans le glycogène et la graisse de ce tissu. L'action de l'insuline liée ou cristalline est également modifiée par l'état de nutrition de l'animal dont le tissu adipeux est utilisé in vitro.

     

Prevalence of anti-HCV and risk factors for hepatitis C virus infection in healthy pregnant women

Summary The prevalence of anti-HCV antibodies and the risk factors for HCV infection were assessed in 5,672 pregnant women living in North Italy. All reactive sera were confirmed by RIBA-2 test. Anti-HCV positive pregnant women together with an anti-HCV negative control group, were interviewed by standardised questionnaire to identify known or potential risk factors for HCV infection. The anti-HCV prevalence was 0.7% (40/5,672), higher than that observed among blood donors in the same geographical area (0.2%). The RIBA-2 assay was positive in 60% (24/40) of cases, indeterminate in 10% (4/40) and negative in 30% (12/40). As for known risk factors, considering RIBA-2 positivity, intravenous drug use was by far the main risk factor for HCV infection, resulting in a significantly higher risk than in the control group (50% versus 5.9% [O. R. 15.8, CI 5.4–45.5]). The ten RIBA-2 positive women without histories of transfusion or IV drug use had a significantly higher frequency of sexual contacts with IV drug users compared to controls (50% vs 4.9% [O. R. 19.0, CI 3.6–94.0]). In conclusion, our study provides evidence that in our geographical area the anti-HCV antibody prevalence is higher in pregnant women than in blood donors and that IV drug use and sexual contacts with IV drug users represent the most important risk factors for HCV infection among young women in North Italy.

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Prävalenz von anti-HCV und Risikofaktoren für eine Hepatitis C Virus-Infektion bei gesunden Schwangeren

Zusammenfassung Bei 5672 Schwangeren, die in Norditalien leben, wurden anti-HCV Prävalenz und Risikofaktoren für eine HCV-Infektion bestimmt. Alle reaktiven Seren wurden mit dem RIBA-2-Test nachgetestet. Anti-HCV positive Frauen und eine anti-HCV-negative Kontrollgruppe wurden mit einem Standardfragebogen interviewt, um bekannte oder mögliche Risikofaktoren für eine HCV-Infektion zu identifizieren. Die Prävalenz von anti-HCV-Antikörpern lag bei 0,7% (40/5672) und war höher als bei Blutspendern aus derselben Gegend (0,2%). Bei 60% (24/40) war der RIBA-2-Test positiv, bei 10% (4/40) unbestimmt und bei 30% (12/40) negativ. Bezogen auf RIBA-2 positive Fälle fand sich unter den bekannten Riskofaktoren intravenöser Drogengebrauch als Hauptrisikofaktor, wobei das Risiko signifikant höher war als in der Kontrollgruppe (50% versus 5,9%; OR 15,8; CI 5,4–45,4). Die zehn RIBA-2 positiven Frauen ohne Vorgeschichte einer Transfusion oder i.v. Drogenmißbrauchs hatten signifikant häufiger sexuelle Kontakte mit i.v. Drogenabhängigen als Kontrollen (50% gegenüber 4,9%; OR 19,0, CI 3,6–94,0). Aus den Daten ist zu schließen, daß die Prävalenz von anti-HCV-Antikörpern bei schwangeren Frauen höher ist als bei Blutspendern und daß intravenöser Drogengebrauch und sexuelle Kontakte mit intravenös Drogenabhängigen die wichtigsten Risikofaktoren für eine HCV-Infektion bei jungen Frauen in Norditalien sind.

     

Local and Systemic Liberation of Proinflammatory Cytokines in Ulcerative Colitis

Determination of plasma and tissue cytokinelevels in inflammatory bowel disease have frequentlyresulted in conflicting data. In the present study wedetermined in patients with ulcerative colitis (UC), the levels of the proinflammatory cytokinesinterleukin (IL)-1, IL-6, interferon(IFN)-, and tumor-necrosis factor (TNF)-liberated by peripheral blood mononuclear cells (PBMC)and lamina propria mononuclear cells (LPMC) after 48-hrculture with pokeweed mitogen (PWM). IL-1, IL-6,IFN- and TNF- in the supernatant weredetected by ELISA. Results show low basal levels ofIL-1 secretion by PBMC and LPMC, and a considerableincrease after mitogen stimulation. Basal IL-6production by PBMC was higher in UC patients than incontrols [2029 pg/ml, CI₉ (–165 to4223) vs 572 pg/ml (–383 to 1527) respectively, P = 0.05] and also afterPWM activation [14,995 pg/ml (7759 -22230) vs 6598 pg/ml(3240-9956), respectively, P = 0.05]. In LPMC, nodifferences in IL-6 secretion were observed. TNF- in activated PBMC of patients with UC was notsignificantly increased in relation to control (P =0.09). No constitutive secretion of IFN- wasobserved in mononuclear cells. IFN- levelssecreted by activated LPMC were lower in patients withUC than in controls [1571 pg/ml (–108 to 3251) vs7953 pg/ml (3851-12,055), respectively, P = 0.03]. Theseresults suggest that IL-6, IL-1, and TNF- participate as mediators in the inflammatoryphenomena observed in UC. Further studies are necessaryto evaluate the role of IFN- in thiscondition.     

Sensitivity of Ph1+ CFU-GM to human recombinant interferon α and γ alone and in combination

Summary The in vitro effect of human recombinant interferon (IFN) alone and in combination were studied on granulomonocytic colony forming units (CFU-GM) from the peripheral blood of 10 Ph1+ chronic myeloid leukemia (CML) patients and from the marrow of 5 normal or non-leukemic subjects. - and -IFN alone determined a slight inhibition on colony growth with a preferential effect on pure macrophagic colonies. At maximum concentration (10⁴ U/ml) leukemic colony inhibition was 46±34% for IFN and 43±19% for IFN. Culture growth with + IFN in combination were significantly inhibited (up to 96±4%) with a concentration-related effect. Similar results were obtained with normal CFU-GM. The synergism that was found in vitro is probably relevant for the in vivo therapeutic effects of these compounds in CML and suggest that the combination is worth testing in vivo.This work was supported by CNR, Oncology Finalized Project, contract n. 8701169.44. and n. 8600603.44. and by AIRC     

Protein kinase C expression in salivary gland acinar epithelial cells in non-obese diabetic mice,an experimental model for Sjögren''s syndrome

We planned to investigate the expression of protein kinase C (PKC) isoforms in acinar epithelial cells of salivary glands in the non-obese diabetic (NOD) mouse to find out if they develop changes of the PKC system like those seen in the human counterpart, i.e. in Sjögren's syndrome. Parotid, submandibular, and sublingual glands from NOD and control BALB/c mice were stained with a panel of monoclonal antibodies directed against conventional (, , and ), novel (, , and ), and atypical ( and ) PKC isoforms using the streptavidin/HRP method. Similarly to human labial salivary glands, acinar epithelial cells of the healthy control BALB/c mice contained two of the conventional PKC isoforms, and . Acinar and ductal epithelial cells also contained the atypical PKC isoforms and . PKC isoforms , , , and were not found. NOD mice which displayed focal sialadenitis contained the same conventional and atypical PKC isoforms. The acinar cells in NOD mice, in contrast to the Sjögren's syndrome patients, did not lack PKC or . On the contrary, PKC and staining was stronger than in the control BALB/c mice. The present results demonstrate that both conventional and atypical PKC isoforms participate in the salivary epithelial cell biology and that there are mouse strain-associated and/or disease state-associated changes in their expression. The lack of PKC and isoforms found in Sjögren's syndrome was not reproduced in NOD mice, which discloses one more difference between the human disease and its NOD mouse model.     

Lavage Administration of Dilute Surfactant in a Piglet Model of Meconium Aspiration

Maldistribution of exogenous surfactant may preclude any clinical response in acute lung injury associated with surfactant dysfunction. Our previous studies have shown the effectiveness of surfactant lavage after homogenous lung injury. The present study utilizes a histologically confirmed non-homogeneous lung injury model induced by saline lung-lavage followed by meconium injected into a mainstem bronchus. Piglets were then treated with Infasurf® or Exosurf® by lavage (I-LAVAGE, n=7; E-LAVAGE, n=5) or bolus (I-BOLUS, n=8; E-BOLUS, n=5), or went untreated (CONTROL, n=4). Lavage administration utilized a dilute surfactant (35 ml/kg; 4 mg phospholipid/ml) instilled into the lung, followed by gravity drainage. The retained doses of the respective surfactant in the lavage and bolus groups were similar. Results showed that the surfactant distribution was more uniform in the lavage groups compared to the bolus groups. Significant and consistent increases in PaO₂ were observed in the lavage groups compared to the bolus groups and the controls. PaO₂ (mmHg) at 240 min posttreatment: I-LAVAGE=297±54, E-LAVAGE= 280±57; I-BOLUS=139±31; E-BOLUS=152±29; C=119±73 (mean± SEM). Other improved pulmonary function parameters favored lavage administration. We conclude that better surfactant distribution achieved by lavage administration can be more effective than bolus administration in this type of non-homogeneous lung injury.     

Zur Rolle der enzymatischen DNA-Reparatur bei Carcinogenese und Krebschemotherapie

Ohne ZusammenfassungCancer Research and Clinical Oncology publishes in loose succession Editorials and Guest Editorials on actual and/or controversial problems in experimental and clinical oncology. These contributions represent exclusively the personal opinion of the author. The EditorsDie Zeitschrift für Krebsforschung und Klinische Onkologie bringt in zwangloser Folge Editorials zu aktuellen und/oder kontroversen Problemen der experimentellen und klinischen Onkologie. Diese Beiträge geben ausschließlich die persönliche Meinung des Autos wieder. Die HerausgeberHerrn Prof. Harold P. Rusch, Madison, Wisc., in aufrichtiger Verehrung zum 70. Geburtstag gewidmet     

Inhibition of protein kinase C in multidrug-resistant cells by modulators of multidrug resistance

We have evaluated the protein kinase C (PKC) activity in two series of cultured cell lines presenting the multidrug-resistance (MDR) phenotype and in the corresponding wild-type cells: the human KB 3.1, KB A1 and KB 8.5 cell lines, and the rat C6, C6 0.5 and C6 1V cell lines. We have observed an increase in PKC activity in the MDR cell lines of the KB cell lineage, proportional to their degree of resistance to doxorubicin. In contrast, the MDR cell lines of the C6 cell lineage presented no change (C6 0.5) or even decrease (C6 1V) in PKC activity; the basal level of PKC activity in C6 cells was, however, 50-fold higher than in KB 3.1 cells. We have tested, in these lines, the effect of four modulators of MDR: verapamil, cyclosporin A, quinine and S-9788, on doxorubicin acytotoxicity and on PKC activity. We observed that cyclosporin A and S-9788, which were the most active on MDR reversal, were able to inhibit PKC activity in the KB resistant lines as well as in all C6 lines, whereas verapamil and quinine had only marginal effects on PKC activity. The distribution of PKC isoenzymes was studied by Western blots. The PKC , and isoforms were increased in the KB resistant lines as compared to wild-type cells, which could account for the increase PKC activity we observed. In contrast, PKC and were decreased in C6 1V cells, as expected from the results obtained for total PKC activity, but we also noticed an important decrease in PKC in the C6 0.5 line. Our results suggest that an increase in PKC activity is not an absolute requirement for expression of MDR, provided that the basal level be high enough; and that some modulators may act on MDR, not only through direct P-glycoprotein interaction, but also through P-glycoprotein phosphorylation or expression. The distribution of PKC isoenzymes revealed that the modifications encountered between sensitive and resistant cells mainly concerned , and isoenzymes of PKC.Abbreviations PKC protein kinase C - MDR multidrug resistance