Effects of bile salts on plasma concentration of β-endorphin-like immunoreactivity in men

The effects of bile salts on the release of -endorphin-like immunoreactivity ( -END-LI) were investigated in men using a specific radioimmunoassay developed by the authors. Plasma -END-LI was determined after extraction by the acid-acetone method (recovery: 73±5%). Oral administration of 400 mg of sodium taurocholate caused a rise in plasma -END-LI from 9.9±0.5 pmol/liter to 21.3±1.2 pmol/liter after 30 min and 18.1 ±0.5 pmol/liter after 60 min, with return to the initial value after 90 min. Oral administration of chenodeoxycholic acid (CDCA) also increased plasma -END-LI from a basal level of 8.4±0.7 pmol/liter to 18.7±0.8 pmol/liter after 30 min. Oral administration of ursodeoxycholic acid (UDCA) increased plasma -END-LI from 7.3±0.3 pmol/liter to 30.6±0.2 pmol/liter after 30 min. In gel chromatography, the -END-LI released after UDCA administration separated into two components, which eluted in the same positions as human -lipotropin and human -endorphin, respectively. These results suggested that bile salts may participate the release of -END-LI.     

Modulation of IL-1β, IL-6, IL-8, TNF-α, and TGF-β secretions by alveolar macrophages under NO2 exposure

Activated alveolar macrophages (AMs) secrete interleukine (IL)1, IL-6, IL-8, tumor necrosis factor- (TNF-), and transforming growth factor- (TGF-), whose inflammatory and fibroblast-activating characteristics may play a role in the maintenance of pulmonary inflammatory processes and subsequent fibrosis. Human AMs were transferred to a gas cylinder and exposed to NO₂ in concentrations ranging from 0.1 to 0.5 ppm in synthetic air for 30 min at 37°C. AMs were fixed on a polycarbonate membrane and placed on culture medium. A culture was established, with the exposed AM (nonstimulated or stimulated with 1 g/ml lipopolysaccharide [LPS]), and the remaining cells were used to determine the cytokines. IL-1, IL-6, and IL-8 were quantified by commercial enzyme-linked immunosorbent assay kits (ELISA kits). TNF- was determined with a sandwich ELISA, using the biotin-streptavidin system. NO₂ exposure of nonstimulated AM did not result in changes in IL-1, IL-6, TNF-, and TGF- release, compared to the situation with control experiments. Exposure for 30 min to NO₂ induced a significant decrease of LPS-stimulated IL-1, IL-6, IL-8, and TNF- (p < .05). The release of TGF- was not significantly affected by NO₂ exposure. Cytotoxicity of AM was checked by trypan blue exclusion, with values ranging from 1.3 to 3.0%. NO₂ exposure of LPS-stimulated AM resulted in a functional impairment of AM after NO₂ exposure regarding IL-1, IL-6, IL-8, and TNF-. Neither the spontaneous nor the stimulated release of TGF- were influenced by NO₂.     

Serum cytokines in patients with rheumatoid arthritis

Serum cytokines such as interleukin 1 (IL-1), interferon (IFN-), and tumor necrosis factor (TNF) were measured in 40 patients with rheumatoid arthritis (RA). In the 40 patients studied, serum IL-1 was detected in 5 patients, IFN- in 10 patients, and TNF in 20 patients. The IL-1-positive group showed increased values of activity indices compared to the IL-1-negative group. Values of serum IFN- correlated well with the number of peripheral blood lymphocytes and CD3⁺ cells and with the percentage of CD3⁺ CD26⁺ cells. Values of serum TNF correlated positively with the number of peripheral blood monocytes and the percentage of CD3⁺ HLA-DR⁺ and CD3⁺ CD25⁺ cells. These results indicated that serum IL-1 in RA patients reflects the activity of RA, while the serum IFN- and TNF in RA patients may be related to circulating activated lymphocytes and monocytes, respectively.     

Die β-Laktamase-Inhibitoren und ihre klinische Bedeutung

Zusammenfassung Zu den verschiedenen Möglichkeiten der Überwindung einer -Laktamase-bedingten Resistenz von Mikroorganismen gehört der Einsatz von Enzyminhibitoren, die selbst keine nennenswerte eigene antimikrobielle Aktivität aufweisen, jedoch in Kombination mit einem Breitspektrumantibiotikum vom -Laktamtyp synergistisch wirken. Auf diese Weise gelangen -Laktam-resistente Bakterien erneut in das Wirkungsspektrum von Substanzen wie Penicillin G oder Ampicillin, die aufgrund steigender Resistenzentwicklung in den letzten Jahren ihre therapeutische Effizienz zu verlieren drohen. 6--Bromopenicillansäure und die sogenannten Olivansäuren weisen eine bemerkenswerte Hemmpotenz gegenüber verschiedenen -Laktamasen auf. Die mikrobiologischen und bisher vorliegenden pharmakokinetischen Daten eines Penicillansäuresulfons, das ebenfalls signifikante Hemmeigenschaften verschiedener klinisch relevanter -Laktamasen besitzt, werden diskutiert. Von Clavulansäure, einem Stoffwechselprodukt vonStreptomyces clavuligerus mit -Laktamstruktur konnte ebenfalls gezeigt werden, daß es ein progressiver Hemmstoff der -Laktamasen vom Richmond-Typ II-V ist. Neben den bisher vorliegendenIn-vitro-Untersuchungen werden auch Ergebnisse klinischer Studien mit der Kombination Clavulansäure und Amoxicillin erwähnt.

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Inhibitors of -lactamases and their clinical significance

Summary One of the various possibilities of overcoming bacterial resistance due to -lactamase production is with enzyme inhibitors. These have no remarkable intrinsic antimicrobial activity, but act synergistically in combination with a broad spectrum antibiotic of the -lactam type. Thus -lactam resistant bacteria are once again within the antibacterial spectrum of substances like penicillin G or ampicillin, which have been in danger of losing their therapeutical effectiveness in recent years due to an ever increasing development of resistance. 6--bromopenicillanic acid and the so-called olivanic acids exhibit remarkable inhibitory properties against several -lactamases. Microbiological and pharmacokinetic data published recently on a penicillanic acid sulfone, which also shows significant inhibitory properties against several clinically relevant -lactamases, are discussed in this paper. Clavulanic acid, a recently discovered product ofStreptomyces clavuligerus with a -lactam structure, acts as a progressive inhibitor of Richmond type II-V -lactamases. In addition to microbiological and enzyme-kineticin vitro data, results of clinical studies with the combination clavulanic acid and amoxicillin are summarized.

     

Serum zinc and copper in active rheumatoid arthritis: Correlation with interleukin 1β and tumour necrosis factor α

Serum zinc and copper levels and serum interleukin 1 (IL1) and tumour necrosis factor (TNF) levels were evaluated in 57 female patients with active rheumatoid arthritis (RA) to investigate a possible role of IL1 and TNF on zinc and copper homeostasis in RA. Serum zinc levels were significantly lower and serum copper levels significantly higher in RA patients when compared with osteoarthritis or asymmetrical psoriatic oligoarthritis patients and with normal controls. No differences were observed in serum IgM rheumatoid factor positive and serum IgM rheumatoid factor negative patients as regards serum zinc and copper concentration. In RA patients the erythrocyte sedimentation rate and acute-phase proteins correlated negatively with serum zinc and positively with serum copper. IL1 and TNF were found to correlate negatively with zinc and positively with copper in RA patients. Lower levels of zinc may be due to an accumulation of zinc-containing proteins in the liver and in the inflamed joints in RA. Elevated serum copper levels seem to be linked to the increased synthesis of ceruloplasmin by the liver.     

Detection of anti-bovine β2-glycoprotein I antibody in sera from patients with antiphospholipid syndrome

We studied and characterized anti-bovine ₂ I antibodies (aB₂-GPI) in sera from patients with antiphospholipid syndrome (APS) by ELISA. Bovine ₂-glycoprotein I ₂-GPI was purified by heparin affinity and DEAE ion-exchange chromatography, and identified on immunoblots using a monoclonal antibody against human ₂-GPI and by amino acid sequence analysis. aB₂-GPI levels in the sera from 36 APS patients were measured by ELISA using purified bovine ₂-GPI as an antigen. The mean±standard deviation level of aB₂-GPI was 17.4±22.0 units in the 58% of APS patients who were positive. There was a significant correlation (P=0.003) between aB₂-GPI and anticardiolipin antibody (aCL) levels. aB₂-GPI from the sera of patients with APS was inhibited by bovine ₂-GPI itself. Purified IgG from the sera of patients with APS showed that bovine ₂-GPI was capable of acting as a cofactor for aCL. Purified bovine ₂-GPI was useful antigen for conventional ELISA. aB₂-GPI may contribute to the further development of aCL analysis and to the understanding of the pathogenesis of APS.     

Oligosaccharides accumulated in the bovineβ-mannosidosis kidney

Summary The phenotype of bovine-mannosidosis (-mannosidase deficiency), recently identified in Salers cattle, is similar to the caprine form of the disease (Abbittet al., 1991). This investigation was designed to characterize accumulated kidney oligosaccharides in bovine-mannosidosis. Oligosaccharides were extracted from the kidney of an affected Salers calf and purified by chromatographic techniques. The amount of accumulating oligosaccharides in 1 g of wet tissue was about 21µmol. Structures of derivatized oligosaccharides were characterized by high-performance liquid chromatography, mass spectrometry, methylation analysis and sequential exoglycosidase digestions. The major accumulating oligosaccharides were Man1-4GlcNAc and Man1-4GlcNAc1-4GlcNAc. Oligosaccharides accumulating in minor amounts were Man1-4GlcNAc1-4Man1-4GlcNAc, Man1-6Man1-4GlcNAc1-4GlcNAc and Man1-4GlcNAc1-4Man1-4GlcNAc1-4GlcNAc. As in caprine-mannosidosis, oligosaccharides with terminal-mannose residues and cleaved as well as uncleaved chitobiose linkages were identified in bovine-mannosidosis kidney. The accumulating oligosaccharides in tissue were thus identical in bovine and caprine-mannosidosis; however, the source of the novel oligosaccharides remains to be determined.     

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.     

IL-1β and IL-6 in community-acquired pneumonia: Bacteremic pneumococcal pneumonia versusMycoplasma pneumoniae pneumonia

Summary Interleukin-1 (IL-1) and interleukin-6 (IL-6) levels in 20 patients with bacteremicStreptococcus pneumoniae community-acquired pneumonia (CAP) were compared with these cytokine levels in 20 patients withMycoplasma pneumoniae CAP. All 40 patients survived hospitalization and underwent a follow-up examination one month later. Serum IL-1 and IL-6 levels were determined by the enzyme immunoassay (EIA) method using commercial kits. In the acute phase of CAP, IL-6 levels were significantly higher in theS. pneumoniae group (p=0.014), while IL-1 levels were higher in theM. pneumoniae group (p=0.046). In the convalescence phase, the two cytokines were detected in a considerable number of patients in both groups. In this phase, only the level of IL-1 was significantly higher in theM. pneumoniae group than in theS. pneumoniae group (p=0.03). We conclude that the levels of IL-1 and IL-6 are different between patients withS. pneumoniae-CAP andM. pneumoniae-CAP during the acute phase. In the convalescence phase, cytokine levels remain high in some of the CAP patients, but a significant difference between the groups exists only for IL-1. Further studies are required.     

Increased levels of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) in tracheal aspirates of newborns with pneumonia

Summary Pneumonia is one of the major sites of infection in ventilated newborns. We investigated whether the cytokines IL-1 and TNF- are detectable in tracheal aspirates of newborns with pneumonia as a diagnostic marker. All 12 infants with pneumonia had elevated levels of IL-1 (range 30–300 pg/ml) and TNF- (range 60–680 pg/ml), whereas control infants (n=21; respiratory distress syndrome, very low birth weight or infants intubated preoperatively) had no detectable levels of IL-1 or TNF-.In vitro investigations with mononuclear cells of umbilical cord blood were performed to rule out that exogenously added surfactant influences IL-1 and TNF- production. It is concluded that IL-1 and TNF- are important and specific mediators of neonatal pneumonia which may be of diagnostic importance.

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Erhöhte Spiegel von TNF- und IL-1 im Trachealsekret von Neugeborenen mit Pneumonie

Zusammenfassung Die Pneumonie ist eine der Hauptlokalisationen von Infektionen bei Neugeborenen. Wir untersuchten, inwieweit die Zytokine IL-1 und TNF- im Trachealsekret von Neugeborenen mit Pneumonie als diagnostischer Marker nachgewiesen werden können. Alle 12 Kinder mit Pneumonie hatten erhöhte Spiegel für IL-1 (30–300 pg/ml) und TNF- (60–680 pg/ml), während die Kontrollen (n=21; Atemnotsyndrom, sehr untergewichtige Neugeborene, präoperativ intubierte Kinder) keine nachweisbaren Spiegel für IL-1 und TNF- hatten. Um auszuschließen, daß exogen appliziertes Surfactant die IL-1 und TNF--Produktion beeinflußt, wurdenIn vitro-Untersuchungen mit mononukleären Zellen von Nabelschnurblut durchgeführt. Wir schließen aus den Ergebnissen, daß IL-1 und TNF- wichtige und spezifische Mediatoren der Neugeborenenpneumonie sind, die von diagnostischer Bedeutung sein können.

     

Distribution of β-Adrenoceptor Subtypes in Gastrointestinal Tract of Nondiabetic and Diabetic BB Rats A Longitudinal Study

The effects of aging and diabetes on thedistribution of -adrenoceptor subtypes in the gutwere investigated in the BB rat.[¹²⁵I]Cyanopindolol binding to 10-msections was evaluated using film autoradiography. Cyanopindolol binding to -,₁-, and₂-adrenoceptors was displaced by 1M propranolol, 50 nM ICI-89-406, and 100 nMICI-118-551, respectively. -Adrenoceptor bindingwas highest in the circular muscle of proximal colon and lowest in thepylorus of 4- to 5-month-old rats. Aging (8- to10-month-old vs. 4- to 5-month-old rats) was associatedwith increased -adrenoceptor binding in thepylorus and reduced binding in the proximal colon.Diabetes had a time-dependent effect on the level of-adrenoceptor binding. It was increased in theantral and pyloric stomach but longer periods ofdiabetes caused a reduction in -adrenoceptorbinding in the pylorus. Those in the intestine werereduced time-dependently and involved₁- or ₂-adrenoceptorsor both.     

Inhibitory effects of adhesion oligopeptides on the invasion of squamous carcinoma cells with special reference to implication of αv integrins

We studied invasion-related adhesion events in vitro using three squamous carcinoma cell lines (HSC-3, poorly differentiated type; OSC-19, well-differentiated type; and KB cells, undifferentiated type). An in vitro invasion assay through matrigel in the transwell chamber revealed that HSC-3 cells were most invasive, OSC-19 cells moderately invasive and KB cells least invasive. Inhibition assay of invasion using synthetic peptides RGD, RGDV, RGDS, RGDT, IKVAV and YIGSR, showed that invasion of the three cell lines was significantly inhibited by RGDV. There were other peptides that inhibited invasion significantly including IKVAV for HSC-3, and RGDS and YIGSR for OSC-19. HSC-3 cells and OSC-19 cells adhered to fibronectin, laminin, vitronectin, and type IV collagen, and KB cells did not adhere to laminin but did to fibronectin, vitronectin and collagen type IV. Pretreatment of cells with RGDV peptide in the attachment assay reduced the ability of these cells to bind to vitronectin and fibronectin more efficiently than pretreatment with RGDS. Anti-v antibodies inhibited adhesion of HSC-3, OSC-19 and KB cells to vitronectin, but anti-1 antibodies did not inhibit adhesion. Immunofluorescent microscopic examinations showed that all cell lines were positive for anti-5 and anti-v antibodies, and only HSC-3 cells were positive for anti-3 antibody. 51 was not clearly demonstrated in any of the cell lines. RGDV was the most effective inhibitor of squamous cell carcinoma invasion among the synthetic oligopeptides used in this experiment, and it is suggested that it affects v3-and/or v5-mediated carcinoma cell invasion.Abbreviations BSA bovine serum albumin - MEM Eagle's minimal essential medium - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide - PBS phosphatebuffered saline - FITC fluorescein isothiocyanate This work supported in part by a grant from the Osaka Cancer Research Foundation     

Detection of β-ureidopropionase deficiency with HPLC–electrospray tandem mass spectrometry and confirmation of the defect at the enzyme level

The pyrimidine bases uracil and thymine are degraded via the consecutive action of three enzymes to -alanine and -aminoisobutyric acid, respectively. To date, a number of patients have been described with a deficiency of dihydropyrimidine dehydrogenase and dihydropyrimidinase, the first two enzymes of the pyrimidine degradation pathway. In this study, we demonstrate that the first patient presenting with N-carbamyl--amino aciduria, due to a deficiency of -ureidopropionase, was easily diagnosed at the metabolite level using HPLC–tandem mass spectrometry. Urinary analysis showed strongly elevated levels of N-carbamyl--alanine and N-carbamyl--aminoisobutyric acid, with normal or moderately increased levels of the pyrimidine bases and the dihydropyrimidines, respectively. The deficiency of -ureidopropionase was confirmed by measuring all three enzymes of the pyrimidine degradation pathway. No activity of -ureidopropionase could be detected in a liver biopsy of the patient, while a normal activity of dihydropyrimidine dehydrogenase and dihydropyrimidinase was present. Thus, HPLC–tandem mass specrometry proved to be a powerful tool for the initial diagnosis of patients with deficiency of -ureidopropionase.     

Urine- and serumβ2-microglobulin in patients with rheumatoid arthritis: A study of 101 patients without signs of kidney disease

Summary Serum levels and 24-hour urinary excretion of 2-Microglobulin (2-M) was investigated in a group of 101 patients with rheumatoid arthritis (RA), without any other signs of renal disease in past or present, andin a comparable control group. Elevated 24-hour urinary 2-M excretion, due to renal proximal tubules dysfunction, was observed in 19% of the patients and not in the control group. There was a significant correlation with clinical signs of extra-articular RA. It is postulated that the observed increase may be an early symptom of renal involvement in RA. Elevated serum 2 levels, corrected for glomerular filtration rate, were observed in 44% of the RA patients and only in 3% of the control group and correlated with clinical signs of a more severe RA, as well as with increased 24-hour urinary 2-M excretion.     

Interleukin-1β gene polymorphisms in Taiwanese patients with gout

The purpose of this study was to examine whether interleukin-1 beta (IL-1) promoter and exon 5 gene polymorphisms are markers of susceptibility or clinical manifestations in Taiwanese patients with gout. The study included 196 patients in addition to 103 unrelated healthy control subjects living in central Taiwan. From genomic DNA, polymorphisms of the gene for IL-1 promoter and IL-1 exon 5 were typed. Allelic frequencies were compared between the two groups, and the relationship between allelic frequencies and clinical manifestations of gout was evaluated. No significant differences were observed in the allelic frequencies of the IL-1 promoter between patients with gout and healthy control subjects. Additionally, we did not detect any association of the IL-1 promoter genotype with the clinical and laboratory profiles of gout patients. However, there was a significant difference between the two groups in terms of hypertriglyceridemia (P=0.0004, ²=12.52, OR 7.14, 95%CI 0.012–0.22). There was also a significant difference in the genotype of IL-1 exon 5 polymorphism between patients with and without hypertriglyceridemia. Results of the present study suggest that polymorphisms of the IL-1 promoter and IL-1 exon 5 are not related to gout patients in central Taiwan.     

Production of interleukin-1β and tumour necrosis factor-α in patients with benign or malignant ovarian tumours

Summary To assess the role of interleukin-1 (IL-1) and tumour necrosis factor (TNF) in the physiological host defence mechanisms against malignancies, the production of these cytokines in sera, ascitic and cyst fluids and in the tumour tissues of patients with benign or malignant ovarian tumours was studied. IL-1 was found neither in the sera nor in the ascitic fluids of these patients. It was also virtually absent from the cyst fluid samples. However, a mean value of 790 pg IL-1/g tumour was found. Like IL-1, TNF was virtually absent in the serum samples. It was, however, detectable in the ascitic and cyst fluids and tumour tissues. The TNF concentrations were highest in the tumour tissues, with a mean level of 328 pg/g tumour. When comparing the level of IL-1 and TNF in patients with benign tumours to that seen in patients with malignant tumours, no differences in production were observed, regardless of the origin of the test samples. Our results indicate the production of IL-1 and TNF in patients with ovarian tumours. More importantly, the finding that the production of these cytokines in patients with benign tumours is similar to that in patients with malignant tumours supports the conclusion that the production of these cytokines is more a nonspecific indicator of an inflammatory process than a specific response to a malignant process.Abbreviations IL interleukin - TNF tumour necrosis factor     

Presence of Two Signaling TGF-β Receptors in Human Pancreatic Cancer Correlates with Advanced Tumor Stage

Transforming growth factor- (TGF-)signal transduction is mediated via specific cellsurface signaling TGF- receptors, most notably thetype I ALK5 (TR-I_ALK5)and the type II(TR-II). We evaluated TR-I_ALK5 andTR-II expression in 41 human pancreatic cancertissue samples and correlated these findings withclinical data of the patients. Northern blot analysisindicated that, in comparison with the normal pancreas,pancreatic adenocarcinomas exhibited 8.0-fold and4.5-fold increases (P < 0.01), respectively, in mRNAlevels encoding TR-I_ALK5 andTR-II. In situ hybridization showed that both TR-I_ALK5 mRNAwere highly expressed in the majority of pancreaticcancer cells. Immunohistochemical analysis ofTR-I_ALK5 and TR-II revealedpositive immunostaining in 73% and 56% of the tumors, respectively. Both receptorswere concomitantly present in 54% of the pancreaticcancer samples. The presence ofTR-I_ALK5 or TR-II and theconcomitant presence of TR-I_ALK5 and TR-II in the cancer cells was associatedwith advanced tumor stage (P < 0.01). These findingsshow that in many human pancreatic cancers, increasedlevels of the two signaling TRs are present. The presence of the signaling TRs inadvanced tumor stages indicates a role in diseaseprogression.     

The antigenicity of human hemoglobins

Summary The correlation of the antigenicities among native hemoglobins and their subunit chains were investigated by the absorption of antisera and the combination of urea added immunoelectrophoresis with double diffusion. Alphachain showed identity with Hb-F but partial identity with -chain and Hb-A. Beta-chain showed identity with Hb-A but -chain and Hb-F showed partial identity with this chain. Gamma-chain showed identity only with Hb-F and its antigenicity was considered as being different from those of - or -chains.The lines of -, -and -chains were reconfirmed from the facts that the appearance of them depended always on the existence of anti-Hb-A or anti-Hb-F antibodies in the absorbed antisera and the minor component lines of

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Zusammenfassung Die Zusammenhänge der Antigenität zwischen nativen Hämoglobinen und deren Unterketten wurden mit der Absorption der Antiseren und der Kombination der Harnstoff-Immunelektrophorese und Doppeldiffusion untersucht. Die -Kette zeigte Identität mit Hb-F, aber nur partielle Identität mit der -Kette und Hb-A. Die -Kette war in ihrer Antigenität mit Hb-A identisch, die -Kette und Hb-F waren teilweise identisch mit der -Kette. Die -Kette zeigte die Identität mit Hb-F; es wird angenommen, daß ihre Antigenität verschieden von der -oder -Ketten ist.Für das Auftreten der Linien der -, - und -Ketten müssen Anti-Hb-A-oder Anti-Hb-F-Antikörper in den absorbierten Antiseren vorhanden sein, außerdem fusionieren die schwächeren Linien der Doppeldiffusion nicht mit irgendwelchen Linien der Unterketten. Auch gereinigte - oder -Ketten wurden zur Feststellung ihrer Linien benutzt.

     

Role of β-Adrenergic Receptor Subtypes in Lipolysis

In vitro lipolysis stimulated by low (-)-isopre-naline concentrations (30 nM) in epididymal white adipo-cytes from Sprague-Dawley rats was inhibited at least 60–80% by the specific ₁-antagonists LK 204-545 and CGP 20712A (1 M), suggesting that at these low (10 nM) concentrations of (-)-isoprenaline lipolysis was primarily (80%) but not solely mediated via ₁-adrenergic receptors. Low concentrations (100 nM) of (-)-noradrenaline and formoterol also confirmed a role for ₁-adrenergic receptors in mediating lipolysis at low concentrations of these agonists. At higher agonist concentrations, ₃-adrenergic receptors were fully activated and were the dominant -adrenergic receptor subtype mediating the maximum lipolytic response, and the maximum response was not affected by the ₁-antagonists, demonstrating that the ₃-receptor is capable of inducing maximum lipolysis on its own. Studies of lipolysis induced by the relatively ₂-selective agonist formoterol in the presence of ₁-blockade (1 M CGP 20712A) demonstrated the inability of the ₂-selective antagonist ICI 118-551 to inhibit the residual lipolysis at concentrations of ICI 118-551 1 M. Higher concentrations of ICI 118-551 inhibited the residual formoterol-induced lipolysis competetively, but with low affinity (500-fold lower than its ₂-adrenergic receptor pA ₂, 7.80 ± 0.21), suggesting that formoterol was not acting via ₂-adrenergic receptors. These data are consistent with ₁-adrenergic receptors playing an important role in lipolysis at physiological but not pharmacological concentrations of catecholamines and that ₂-adrenergic receptors play no obvious direct role in mediating -adrenergic receptor agonist-induced lipolysis in vitro. Finally, racemic-SR 59230A, unlike the pure (S, S)-isomer (a ₃-selective antagonist), was found to be a non-selective antagonist at the three -adrenergic receptor subtypes, showing that the other enantiomers have different selectivity.     

β-Endorphin-like immunoreactivity in normal mucosa,muscle layer,adenocarcinoma, and polyp of the colon

-Endorphin-like immunoreactivity was detected in the mucosa and muscle layer of normal colon, adenocarcinomas derived from the colon mucosa, and colon polyps which were histologically confirmed to be adenoma without a focus of carcinoma or with in situ carcinoma. The contents of -endorphin-like immunoreactivity in adenocarcinomatous tissue (11.94± 1.77 pmollg wet wt) and colon polyps without focus of carcinoma (10.71 ±1.50 pmollg wet wt) were found to be significantly higher than those in the mucosal layer (6.86± 0.64 pmollg wet wt) and muscle layer (8.30 ±0.68 pmollg wet wt) of normal colon. These data suggest that the production of -endorphin-like immunoreactivity is specifically increased in some adenocarcinomas and adenomatous polyps and may be related to the alteration of bowel habits. Gel exclusion chromatography of - endorphinlike immunoreactivity revealed three peaks corresponding to -endorphin, -lipotropin, and an immunoreactive form between the two. In the mucosal layer and muscle layer of the colon, a broad major peak was eluted at the position of -endorphin, and minor peaks were eluted at the position of -lipotropin and between -endorphin and -lipotropin. In adenocarcinoma and polyp, the peak size corresponding to authentic -lipotropin was greater than that of -endorphin. This study demonstrated that -endorphin-like immunoreactivity existed at a high concentration in some colon adenocarcinomas and polyps whose elution patterns were different from those of normal colon tissue.