Histochemischer Nachweis der β-Glucuronidase (EC 3.2.1.31) und β-Acetylglucosaminidase (EC 3.2.1.30) in der Rattenleber nach Gabe von 1-Naphthyl-Isothiocyanat und D-Galaktosamin-Hydrochlorid

Zusammenfassung In der vorliegenden Arbeit konnte gezeigt werden, daß die histochemisch nachweisbaren Reaktionsprodukte der-Glucuronidase und-Acetylglucosaminidase der Rattenleber nach Gabe von 1-Naphthyl-isothiocyanat und D-Galaktosamin-HCl zunehmen. Die-Glucuronidase ist vorwiegend in den Kupfferschen Sternzellen nachweisbar.

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Summary In this study the authors, for the first time, could demonstrate by histochemical methods that the activities of-glucuronidase and-acetyl glucosaminidase in rat liver increase in animals which are pretreated with 1-naphthyl-isothiocyanate and galactosamine. Best reaction of -glucuronidase was found in Kupffer cells.

     

Susceptibility ofBacteroides non-fragilis and fusobacteria to amoxicillin,amoxicillin/clavulanate,ticarcillin, ticarcillin/clavulanate,cefoxitin, imipenem and metronidazole

The susceptibility of 234Bacteroides non-fragilis strains and 56 fusobacteria from 12 European centers to amoxicillin, amoxicillin/clavulanate, ticarcillin, ticarcillin/clavulanate, cefoxitin, imipenem and metronidazole was tested and related to -lactamase production. Beta-lactamase production was detected in 42.3 % of theBacteroides strains and 26.8 % of the fusobacteria. The MIC90 of amoxicillin for -lactamase-negative strains was 0.5 µg/ml and the MIC90 of ticarcillin 2.0 µg/ml. In the case of -lactamase-positive strains the MIC90 of amoxicillin (32 µg/ml) and ticarcillin (16 µg/ml) dropped to 1.0 µg/ml upon addition of clavulanate; 65.8 % of these strains were susceptible to amoxicillin and 98.2 % to ticarcillin, but all were susceptible when clavulanate was added. All strains were susceptible to imipenem and metronidazole, and 99.3 % to cefoxitin.     

Mechanism of modulation of single sodium channels from skeletal muscle by the β 1-subunit from rat brain

We studied the molecular mechanism of the rat skeletal muscle -subunit (_I) gating kinetics modulation by the brain ₁-subunit by heterologous expression of single sodium channels from _I and ₁ in Xenopus laevis oocytes. Coexpression of ₁ reduced mean open time at –10 mV to 21% when compared to channels expressed by _I alone. Channels formed by _I exerted multiple openings per depolarization, which occurred in bursts, in contrast to the channels formed by the _I/₁ complex that opened in average only once per depolarizing voltage pulse. Macroscopic current decay (mcd), as evidenced by reconstructed open probability vs. time , was greatly accelerated by ₁, closely resembling mcd of sodium currents from native skeletal muscle. Generally was larger for channels expressed from the pure _I subunit.From our single channel data we conclude that ₁ accelerates the inactivation process of the sodium channel complex.     

Lack of additive effect between mechanisms of resistance to carbapenems and other beta-lactam agents inPseudomonas aeruginosa

Eighty-nine clinical isolates resistant (n=61) or susceptible (n=28) to imipenem and exhibiting the main patterns of susceptibility to other -lactam agents (wild type pattern, penicillinase pattern, constitutive cephalosporinase pattern) were studied in order to investigate (i) the mechanism of resistance involved and (ii) whether resistance to carbapenems affects the level of resistance to other -lactam agents and, conversely, if resistance to other -lactam agents affects the level of resistance to carbapenems. For this purpose, the presence of OprD protein in the cell wall was detected by Western blot and -lactamase activity by spectrophotometric assay and isoelectric focusing. OprD expression was not detectable in the imipenem-resistant (MIC16 g/ml) strains. It was decreased in half the strains for which MICs of imipenem were 2 to 8 g/ml and was close to a normal level in the most susceptible strains (MIC 1 g/ml), thus demonstrating a direct correlation between the level of susceptibility to imipenem and the level of OprD expression. No imipenemase activity was detected in imipenem-resistant strains. Synergy between imipenem or meropenem and BRL42715 was observed for all of the strains, demonstrating the role of cephalosporinase in carbapenem resistance. Within each pattern of susceptibility, the mean MICs of -lactam agents other than carbapenems were similar, whether the strains were susceptible or resistant to imipenem. Conversely, the mean MICs of imipenem or meropenem for either the imipenem-resistant or the imipenem-susceptible strains were similar, regardless of the susceptibility of these strains to the other -lactam agents. Thus, when several mechanisms of resistance to -lactam agents are present in the same strain ofPseudomonas aeruginosa, there is no additive effect between these mechanisms.     

Role of elevated monocyte transforming growth factorβ (TGFβ) production in posttrauma immunosuppression

We previously reported that increased production of prostaglandin E₂ by monocytes is a pivotal mechanism in posttrauma immunopathology. Here we characterize monocyte levels of transforming growth factor and examine the effects of elevated transforming growth factor on prostaglandin E₂ release by patients' monocytes. Trauma patients' and normals' monocyte supernates (± stimulation with muramyl dipeptide) were acid treated and assayed for transforming growth factor using the mink lung-cell bioassay. Alternatively, human transforming growth factor was added to patients' and normals' monocytes and prostaglandin E₂ production assayed. Significantly elevated transforming growth factor levels (median=181.7 pmol/10⁶ monocytes) were detected in immunosuppressed patients' monocytes but not immuno-competent trauma patients' (median=32.0 pM) or normals' (median=20.4 pM) monocytes. Adding transforming growth factor to monocytes resulted in a significant elevation of prostaglandin E₂ levels. Elevated monocyte transforming growth factor levels in trauma patients could be both suppressing T-lymphocyte functions and maintaining elevated monocyte prostaglandin E₂ synthesis.     

Expansion of neopterin and beta2-microglobulin in cerebrospinal fluid reaches maximum levels early and late in the course of human immunodeficiency virus infection

Summary Elevated cerebrospinal fluid (CSF) levels of neopterin and beta₂-microglobulin (\2MG) reflect activation of the cellular immune response in the central nervous system (CNS). In 118 consecutive subjects [15 controls and 103 patients with human immunodeficiency virus (HIV) infection classified according to the Walter Reed staging system (WR)], neopterin and 2MG were determined in paired samples of CSF and serum. The permeability of the blood-CSF barrier and local release of neopterin and 2MG were taken into account: The molecular weight and diameter were used to determine filtration at the blood-CSF barrier. CSF neopterin levels were increased in all stages of HIV infection. 2MG levels were elevated in WR2 and later stages. Neopterin, 2MG, and cell counts similarly showed peaks in WR2, as did neopterin and 2MG also in the later stages WR5 and WR6. Neurologically asymptomatic patients exhibited higher neopterin CSF levels than did controls (12.67 ± 11.6 vs. 2.34 ± 1.05 nmol/l, P < 0.001) and higher CSF 2MG (2.12 ± 1.25 vs. 1.3 ± 0.37 mg/l, P=0.001). Patients with HIV encephalopathy had higher levels of 2MG (3.75 ± 1.83 mg/1) than asymptomatic patients (P < 0.01). CSF levels of neopterin were markedly different in patients with HIV encephalopathy and toxoplasmosis (P < 0.01). A high quantity of local release of the markers neopterin and 2MG may reflect HIV infection of the CNS in early and late stages and additional release upon opportunistic infections.Abbreviations AIDS acquired immunodeficiency syndrome - \2MG beta₂-microglobulin - CNS central nervous system - CSF cerebrospinal fluid - HIV human immunodeficiency virus (type I) - RIA radioimmunoassay - SD standard deviation - SEM standard error of the mean - WR Walter Reed (staging classification) - ELISA enzyme-linked immunosorbent assay     

Regulation of interleukin-1β and tumor necrosis factor secretion by the human monocytic leukemia cell line,THP-1

Activated macrophages synthesize and release the potent polypeptides, interleukin-1 (IL-1) and tumor necrosis factor (TNF). In an effort to identify the cellular signals which control cytokine production by activated macrophages, we have developed anin vitro model employing the human THP-1 cell line. In the present study, THP-1 cells primed by 1.6 M phorbol 12-myristate-13-acetate (TPA) for 4 hr demonstrated a dose-and time-dependent release of IL-1 and TNF upon activation by 20 g/ml LPS. BSA/anti-BSA-coated latex beads were also a potent stimulus for IL-1 secretion. Moreover, the combination of a suboptimal concentration of LPS (200 ng/ml) plus interferon- (0.03–333 U/ml) greatly enhanced IL-1 production. Resting THP-1 monocytes not primed by TPA did not secrete IL-1 or TNF. These distinct patterns of cytokine production may be related to the developmental stages of macrophage activation.     

Interleukin 1β (IL-1β) release from fresh and cultured colonic mucosa in patients with ulcerative colitis (UC)

Interleukin-1, a cytokine produced by macrophages and other tissue cells, has a major role in inflammatory and immunological responses. Increased levels of IL-1 activity have been reported in experimental colitis and in patients with active Crohn's disease (CD) and ulcerative colitis (UC). IL-1 release from fresh and cultured colonic biopsies and IL-1 plasma concentrations was determined in 15 patients with active UC, 16 with UC in remission and 10 normal control subjects. Biopsies, taken at colonoscopy were weighed, washed in 1 ml of 0.9% sodium chloride solution and then cultured for 24 h in 10% fetal calf serum/RPMI. IL-1 activity was determined by ELISA KIT (Cystron Biotechnology) in plasma samples, washing solution and the incubation medium. Very low levels of IL-1 were detected only in 3 plasma samples, all from active patients. Significantly more IL-1 was released from fresh and cultured colonic mucosa obtained from patients with UC in remission compared to normal mucosa (p<0.01). Furthermore, specimens from active UC released significantly more IL-1 than those from patients in remission (p<0.01). In conclusion, IL-1 may play an important role in mediating the inflammatory response in UC.     

Veränderungen des cAMP-Adenylatcyclasesystems am insuffizienten menschlichen Herzen. Konsequenzen für die Therapie mit positiv inotropen Pharmaka

Zusammenfassung Aufgrund experimenteller Untersuchungen an explantierten menschlichen Herzen lassen sich zwei biochemische Veränderungen charakterisieren: In Abhängigkeit vom Schweregrad der Herzinsuffizienz entsteht eine Abnahme von kardialen ₁-Adrenozeptoren, wohingegen die ₂-Adrenozeptordichte unbeeinflußt bleibt. Zusätzlich kommt es bei der dilatativen Kardiomyopathie zu einer Zunahme von G_i. Diese Veränderungen im-Adrenozeptor-cAMP-Adenylatcyclasesystem führen zu einer Verminderung der Wirkung cAMP-abhängig positiv inotrop wirksamer Substanzen wie-Adrenozeptor-Agonisten und Phosphodiesterasehemmstoffen. Der positive inotrope Effekt von cAMP-unabhängig wirksamen positiv inotropen Maßnahmen wie die Erhöhung der extrazellulären Calciumkonzentration, Aktivierung des Natriumeinwärtsstromes und Hemmung der Natrium-Kalium-ATPase durch Herzglykoside ist bei der Herzinsuffizienz unvermindert. Das bedeutet, daß am insuffizienten menschlichen Herzen die kontraktile Reserve des Myokards im wesentlichen erhalten bleibt, aber ihre Ausschöpfung durch endogene Katecholamine und exogene, cAMP-abhängige positiv inotrop wirkende Pharmaka vermindert ist. Die Konsequenz aus diesen Beobachtungen ist, daß eine pharmakologische Erhöhung der zellulären cAMP-Spiegel durch-Adrenozeptoragonisten oder Phosphodiesterasehemmstoffe bei Herzinsuffizienz nicht sinnvoll erscheint. Eine Kontraktionskraftsteigerung kann am besten durch cAMP-unabhängig wirksame positiv inotrope Substanzen erreicht werden.

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Alterations of the cAMP-adenylate cyclase system in the failing human heart. Consequences for therapy with positive inotropic agents

Summary In heart failure, an increase in the activity of the sympathetic nervous system takes place to maintain perfusion pressure to vital organs, resulting in increased levels of noradrenaline in the blood of these patients. This permanent stimulation produces a down-regulation of cardiac-adrenoceptors. Since noradrenaline acts primarily on the cardiac ₁-adrenoceptor subtype, ₁-adrenoceptors decrease in number, whereas the ₂-adrenoceptor subpopulation remains unchanged in most instances. Consequently, the positive inotropic response to-adrenoceptor agonists is diminished. However, there is also a decrease in the positive inotropic effect of ₂-adrenoceptor agonists, histamine and cAMP-phosphodiesterase inhibitors such as milrinone, whereas the positive inotropic effect of cAMP-independent Na⁺ -channel activators such as DPI 206-106 and the effects of cardiac glycosides are not diminished. These observations suggest a more generalised alteration of the cAMP-adenylate cyclase system in the failing heart. Stimulatory guanine nucleotide-binding protein (G_s) couples receptors to adenylate cyclase that stimulate cAMP formation, such as-adrenoceptors, histamine receptors and glucagon receptors. In the failing human heart, G_s content has been reported to remain unchanged as compared with that in nonfailing myocardium. However, there is a 35%–40% increase in inhibitory guanine nucleotide-binding proteins (G_i), which are involved in the receptor-mediated inhibition of adenylate cyclase. Taken together, two defects of the cAMP-adenylate cyclase system have been identified: an increase in G_i content and a decrease in the number of-adrenoceptors. Both alterations might contribute to the reduced effectiveness of cAMP-dependent positive inotropic agents. cAMP-independent agents might be more efficacious in mobilizing the contractile reserve of the failing myocardium. Reduction of the permanent-adrenergic stimulation of the failing heart, e.g. by-blockers, might be a promising approach for reversing these alterations of the cAMP-adenylate cyclase system.

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Abkürzungen mN Millinewton - cAMP cyclisches Adenosin-35-monophosphat - ATP Adenosintriphosphat Die eigenen experimentellen Untersuchungen wurden mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt.     

Distinct mechanism of human neuroblastoma cell adhesion to fibronectin

We investigated the adhesion of three morphologically distinct human neuroblastoma cell lines (NCG, GOTO and SK-N-DZ) to intact fibronectin, central cell binding domain fragment (CBF) and CS peptide-IgG conjugates in the fibronectin molecule. Each cell line was found to express different integrin fibronectin receptors ( _{3 1}, _{4 1} and ₅₁), although similarly attached on intact fibronectin. To CBF, NCG attached well, while GOTO moderately and SK-N-DZ poorly attached. Only GOTO adhered to CS1-IgG. RGDS inhibited the spreading of NCG and SK-N-DZ on intact fibronectin, but it barely inhibited that of GOTO. The analysis by fluorescence-activated cell sorting (FACS) revealed that NCG expressed abundant ₃₁ and ₅₁, but little ₄₁, while GOTO expressed a large amount of ₄₁ as well as ₅₁. SK-N-DZ was undetectable in any of these molecules, but expressed _v1, which was identified by immunoprecipitation and immunoblotting. Polyclonal antibody to _v3 inhibited the adhesion of SK-N-DZ but not that of NCG or GOTO on intact fibronectin. These results suggest the existence of a distinct mechanism of cell adhesion to fibronectin among human neuroblastoma cell lines. It remains to be determined if such heterogeneous adhesion properties are related to the unique metastatic character of human neuroblastoma.     

Effects of serine protease inhibitor,tame, on IL-1β in LPS-stimulated human monocytes: Relationship between synthesis and release of a 33-kDa precursor and the 17-kDa biologically active species

LPS stimulation of human monocytes in vitro induced release of the 17-kDa mature IL-1 (mIL-1) but did not result in release of precursor IL-1 (pIL-1). In contrast, the presence of a serine protease inhibitor, N-(p-toluene sulfonyl)-L-arginine methyl ester (TAME; 10 mM) for 6 or 18 h was associated with the LPS-stimulated release of the 33-kDa pIL-1 as well. These effects were initially discerned from observations that the fraction of the total IL-1 produced (as detected by ELISA) that was released from monocytes increased in the presence of TAME, and immunoblot assays confirmed that this fraction was predominantly 33-kDa IL-1. A global decrease in monocyte protein synthesis was also observed after prolonged (18-h) exposure to TAME and was associated with a decrease in IL-1 synthesis, predominantly affecting 31-kDa pIL-1, and a dose-dependent inhibition of TNF- production. Parallel examination of lactate dehydrogenase (LDH) release indicated thatpIL-1 release was unrelated to cell lysis. These results demonstrate that TAME-inhibitable serine proteases are probably involved in the production and eventual proteolysis of the 33-kDa pIL-1 in situ but are probably not mechanistically related to either maturation of the IL-1 molecule or signaling of IL-1 release. IL-1 release appears to be dependent on the amount of total IL-1 synthesized. Serine proteolysis may constitute a degradative pathway for excess precursor, which, if interfered with, could result in release of the higher-molecular-weight forms of IL-1.     

Effects of recombinant human IL-Iβ on production of prostaglandin E2, leukotriene B4, NAG,and superoxide by human synovial cells and chondrocytes

The effects of recombinant human IL-1 on the production of prostaglandin E₂ (PGE₂), leukotriene B₄ (LTB₄),N-acetyl--D-glucosaminidase (NAG), and superoxide by synovial cells and chondrocytes derived from osteoarthritis patients were determined. IL-1 markedly enhanced PGE₂ production in chondrocytes and, to the lesser extent, in synovial cells. Synovial cells and chondrocytes spontaneously released LTB₄ into culture medium and IL-1 significantly inhibited LTB₄ production by these cells. IL-1 significantly suppressed the release of NAG and superoxide by synovial cells, whereas it significantly enhanced the production of NAG and superoxide by chondrocytes. Production of intracellular superoxide dismutase by synovial cells was significantly enhanced on incubation with IL-1, but that of chondrocytes was not altered. IL-6, unlike IL-1, significantly suppressed the production of NAG and superoxide by synovial cells and chondrocytes.These results suggest that IL-1 has differing effects on the release of mediators by synovial cells and chondrocytes and that these cells also vary in their responses to IL-1 and IL-6.     

Phenotypic characteristics of lewis lung carcinoma cells

We studied adhesive properties and physiological activity in vivo of cells from Lewis lung carcinoma and its metastases. These cells differed in tumorogenic activity and metastatic potential in the syngeneic system. In vivo non-metastasizing cells are characterized by a lower content of surface lectins to tetrasaccharides SiaLe^x [Neu5Ac2-3Gal1-4(Fuc1-3) GlcNAc] and SiaLe^a [Neu5Ac2-3Gal1-3(Fuc1-4)GlcNAc] and trisaccharide HSO₃Le^x [HSO₃2-3Gal1-4(Fuc1-3)GlcNAc] compared to cells forming metastases in the syngeneic system. Metastatic cells with low tumorogenic activity weakly expressed lectins to disaccharide ligands 6-SiaLac [Neu5Ac2-6Gal1-4Glc], 6-HSO₃LacNAc, and A-di [GalNAc 1-3Gal] and trisaccharides H-type 1 [Fuc1-2Gal1-3GlcNAc and Le^x [Fuc1-3(Gal 1-4)GlcNAc] compared to cells that initiated tumor formation in the syngeneic system (similarly to transplanted tumors). We hypothesized that cell receptors to these carbohydrate determinates are involved in the development and growth of primary tumors, while lectins to SiaLe^x, SiaLe^a, and HSO₃Le^x play a role in the progress of tumor process and metastasizing.     

In vitro antibacterial activity and beta-lactamase stability of the new carbapenem SM-7338

The in vitro activity of the new carbapenem SM-7338 was tested in comparison with imipenem, ceftazidime, cefotaxime, flomoxef, cefuzonam and cefmetazole against 2850 clinical bacterial isolates. SM-7338 showed good activity against a broad spectrum of grampositive and gram-negative bacteria. SM-7338 was very active against gram-negative bacteria, inhibiting allEnterobacteriaceae, except 25 % ofSerratia marcescens isolates, at a concentration of 0.78 mg/l. SM-7338 inhibited the majority ofPseudomonas spp. at concentrations of 3.13 mg/l, its activity being twofold higher than that of imipenem. However, the activity of SM-7338 against gram-positive cocci was about one-fourth that of imipenem. Against anaerobes, SM-7338 also had the best activity of the -lactams tested. The compound was inactive against methicillin-resistant staphylococci,Enterococcus faecium, Xanthomonas maltophilia andFlavobacterium spp., as were the other -lactams. SM-7338 was quite stable in the presence of various types of -lactamase, but was hydrolyzed byXanthomonas maltophilia -lactamase, as was imipenem. This high degree of stability was responsible for the potent activity of SM-7388 against -lactamase-producing species such asEnterobacter cloacae, Citrobacter freundii andProteus vulgaris. SM-7338 also showed bactericidal activity against clinical isolates at the MIC or at concentrations slightly above the MIC.     

In vitro activity of biapenem against beta-lactamase producingEnterobacteriaceae

The activity of biapenem was compared with that of imipenem and cefotaxime against 108 strains of -lactamase producingEnterobacteriaceae. Biapenem and imipenem were very active, inhibiting 90 % of the strains at a concentration of 0.5 µg/ml. Both carbapenems were very active against plasmidic -lactamase producers, with MIC90s below 1 µg/ml. However, the MIC90 of biapenem for cephalosporinase producers was 1 µg/ml. Against strains producing extended-spectrum -lactamases, biapenem exhibited better activity against TEM-type producers (MIC90 0.25 µg/ml) than against SHV-type producers (MIC90 0.5 µg/ml). Overall, the in vitro antibacterial activity of biapenem is similar to that of imipenem.     

Mapping thyrotropin β subunit gene in man and mouse

Thyrotropin (TSH) is composed of two subunits: and . Previously, we have mapped the TSH gene to human chromosome 6 and mouse chromosome 4. In this study we have located the human TSH gene on chromosome 1 and the mouse TSH gene to chromosome 3. These data suggest that the TSH gene lies in a conserved linkage group with the genes for amylase 1 and 2, nerve growth factor, and the protooncogene Nras.     

Harnsteroidprofile bei Cushing Syndrom und Tumoren der Nebennierenrinde

Summary The analysis of 24-h excretion profiles of urinary steroids in 18 patients suffering from Cushing's syndrome or adrenocortical tumors revealed typical patterns when compared to 37 healthy control persons, 24 patients with obesity, and 6 patients with hirsutism. The validation of eight criteria — increased excretion of free cortisol, 6-hydroxycortisol, 20-dihydrocortisol, 11-hydroxyandrosterone, and 3-hydroxy-5-en steroids, decreased ratio of tetrahydrocortisone (THE) to tetrahydrocortisol (THF), and increased ratios of THF to allotetrahydrocortisol (a-THF) and metabolites of androgens (AM) to metabolites of cortisol (CM) — afforded reliable detection of disorders in steroid biosynthesis. The analysis of urinary steroid profiles can therefore be recommended as a screening procedure in patients with clinical symptoms of disorders in steroid production and/or metabolism.

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Abkürzungen An Androsteron - Aet Aetiocholanolon - AM Androgenmetaboliten: An plus Aet - 11-O-An 11-Ketoandrosteron - 11-OH-An 11-Hydroxyandrosteron - 11-OH-Aet 11-Hydroxyaetiocholanolon - DHEA Dehydroepiandrosteron - 16-OH-DHEA 16-Hydroxydehydroepiandrosteron - 16-OH-DHEA 16-Hydroxydehydroepiandrosteron - A⁵T-16 Androsten-3,16,17-triol - P⁵T Pregnentriol-3-hydroxy-5-en Steroide: DHEA plus Androstendiole plus 16-OH-DHEA plus 16-OH-DHEA plus - P⁵D Pregnendiol plus A⁵T-16 plus 16-Hydroxypregnenolon plus P⁵T - THE Tetrahydrocortison - THF Tetrahydrocortisol - a-THF Allotetrahydrocortisol - CM Cortisolmetaboliten: THE plus THF plus a-THF. - -C,-C Cortole - -CL,-CL Cortolone - 6-OH-F 6-Hydroxycortisol - 20-OH-F 20-Dihydrocortisol - NNR Nebennierenrinde - CRF Corticotropin Releasing Hormon - ACTH Adrenocorticotropes Hormon - CPB Competitive Protein Binding - RIA Radioimmunoassay - HPLC High Pressure Liquid Chromatography - DHEAS Dehydroepiandrosteron-Sulfat. Mit Unterstützung der Deutschen Forschungsgemeinschaft, Ho-471/5-1     

Pregnancy specific β1-glycoprotein and α2-pregnancy associated globulin in tumours of the female genital tract

Summary Tumour tissues obtained from 35 patients with malignancies of the female genital tract were investigated for production of pregnancy specific ₁-glycoprotein (PS₁G) and ₂-pregnancy associated globulin (₂-PAG). PS₁G was found in all five trophoblastic tumours studied and in 10 of the 30 non-trophoblastic cancers. ₂-PAG was not detected in any of the neoplastic tissues.In 18 of the patients with non-trophoblastic tumours PS₁G and ₂-PAG serum concentrations were also determined. No correlation between serum and tissue findings were noted. Thus, PS₁G does not seem to be a valuable serum indicator for monitoring the extent or variation of tumour mass in non-trophoblastic gynecological malignancies. Likewise, ₂-PAG is unlikely to serve as a clinical useful tumour marker in various gynecological cancers.     

Immunohistochemical distribution of S-100 protein and glial fibrillary acidic protein in normal and neoplastic salivary glands

Summary Immunohistochemical localization of S-100 protein, its and subunits, and glial fibrillary acidic protein (GFAP) in normal and neoplastic salivary glands was studied by the peroxidase-antiperoxidase method and immunoblot analysis. Positive immunostaining for S-100 protein was observed in pleomorphic adenoma, adenolymphoma, tubular adenoma, adenoid cystic carcinoma, acinic cell tumour, adenocarcinoma and carcinoma in pleomorphic adenoma. S-100 protein was localized in myoepithelial cells, epithelial cells of intercalated ducts and serous acinar cells of normal salivary gland. Both and subunits of S-100 protein showed almost identical distribution in normal and neoplastic salivary glands, but skeletal muscle cells were -positive/-negative whereas Schwann cells and fat cells were -negative/-positive in the stroma and neighbouring tissue. GFAP was only found in pleomorphic adenoma and its malignant counterpart. Immunoblot analysis showed that the GFAP-related antigen consisted of several polypeptide bands with a molecular weight ranging between 35,000 to 50,000 daltons.     

Neutralization of Transforming Growth Factor β1 Augments Hepatitis C Virus-Specific Cytotoxic T Lymphocyte Induction in Vitro

In hepatitis C virus (HCV) infection, TGF-1 is upregulated in the liver and may be involved in the pathogenesis of chronic liver disease. TGF-1 is also produced by activated T cells and acts as a potent immunosuppressor. The aim of this study was to investigate the roles of TGF-1 in HCV-specific cytotoxic T lymphocyte (CTL) induction and enhance their killer activity by TGF-1 modulation. We generated anti-HCV CTL from peripheral blood mononuclear cells from HLA-A2 patients under stimulation with the HCV-core peptide having the HLA-A2.1 binding motif. The lytic activities of CTL or precursor frequency (CTLpf) generated with or without anti-TGF-p antibody were compared. To optimize the IL-2 dose for CTL induction, low (50 U/ml) and high (500 U/ml) doses were tested and the lytic activities were compared. TGF-1 amounts in the supernatants were assessed by enzyme-linked immunosorbent assay and by their growth inhibitory effect on mink lung epithelial cells. CTL activity was enhanced by anti-TGF- antibody in a dose-dependent manner but CTLpf did not significantly change. A high dose of IL-2 reduced the activity to 45% of that observed with a low dose, whereas TGF-1 increased as the dose of IL-2 increased. Exogenous IL-10 reversed the inhibitory effect of a high dose of IL-2 on the killing activity by reducing TGF-1 mRNA expression in T cells and its production. These results demonstrated that endogenous TGF-1 is an autocrine suppressor in CTL induction in vitro. Therefore, the blockade of endogenous TGF-1 could enhance the killing potential of anti-HCV CTL.