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.     

Granulocyte Elastase in Cirrhotic Patients with Spontaneous Bacterial Peritonitis

Granulocyte elastase (GE) is a powerfulproteolytic enzyme that is released by PMNs whendegranulated in infectious processes. The aim of thisstudy was to measure GE in ascites and plasma ofcirrhotic patients with spontaneous bacterial peritonitis(SBP). We studied 29 cirrhotic patients, 17 of themhaving SBP (group A). Twelve patients with noninfectedascites formed the control group (group B). At the time of diagnosis of SBP, GE levels inascites (183.17 ± 86.11 g/liter) and plasma(114.6 ± 35.99 g/liter) were higher in groupA than in group B (27.41 ± 11.54 g/liter, P< 0.00001 and 82.54 ± 20.52 g/liter, P = 0.01,respectively). Levels of GE in ascites had a high valuefor discriminating between patients with and withoutSBP. In the patients who responded to the initialantibiotic treatment, these values significantly decreasedin ascites (67.69 ± 54.22 g/liter, P = 0.003)and plasma (67 ± 22.39 g/liter, P = 0.01) 48hr after therapy was started, in parallel with thedecrease of PMN in ascites. In patients who did notrespond, the production of GE remained elevated.Patients who developed renal insufficiency following SBPhad more marked elevation of GE in plasma (144.8± 33.43 g/liter) than those with normal renalfunction (99.5 ± 27.53 g/liter, P = 0.02).These results suggest that the measurement of GE may behelpful for the diagnosis of SBP in patients withcirrhosis and for assessing the efficacy of therapy. Inaddition, the release of GE into plasma may contributeto the impairment of renal function that follows SBP insome patients.     

Alterations in the heart sarcolemmal Ca2+ transport activity by some β-adrenergic antagonists

Summary The effects of some -adrenergic antagonists such as acebutolol, propranolol, pindolol and oxprenolol (1–1000 M) were studied on the rat heart sarcolemmal Ca²⁺ transport activities. Pindolol enhanced sarcolemmal ATP-dependent Ca²⁺ binding and Ca²⁺-stimulated ATPase whereas acebutolol had no effect. Both propranolol and oxprenolol had biphasic actions on the sarcolemmal Ca²⁺ pump activities; the lower concentrations (1 and 10 M) were stimulatory, but the higher concentrations (100 and 1000 M) were inhibitory. None of the drugs used in this study had any effect on Mg²⁺ ATPase and non-specific Ca²⁺ binding activities of heart sarcolemma except that 1000 M propranolol decreased Mg²⁺ ATPase activity significantly. Mitochondrial and microsomal ATP-dependent Ca²⁺ binding activities were unaffected by these drugs (1–1000 M), except that 1000 M propranolol was inhibitory. These results suggest differences among various -adrenergic blocking drugs with respect to their actions on sarcolemmal Ca²⁺ pump in the myocardium.     

Einfluß der Prämedikation mit Gibberellinsäure auf das Wachstum des an Mäusen subcutan transplantierten Ehrlichschen Ascitestumors

Zusammenfassung Nach einer vorherigen, 9 Tage dauernden Prämedikation mit GS von verschiedenen Konzentrationen (1000, 100, 10 und 1 g/0,5 ml) wurden den Mäusen am 10. Tag eine Suspension von Ehrlich-Ascites-Tumorzellen subcutan transplantiert. Nach weiteren 22 Tagen wurden die Tiere getötet, und der subcutan wachsende Tumor gewogen. Die Werte der Durchschnittsgewichte der Tumoren zeigten, daß die niedrigsten Konzentrationen der applizierten GS (10 und 1 g) die höchste biologische Aktivität ausübten (p=0,001).

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Influence of premedication with gibberellic acid on the growth of subcutaneously transplanted Ehrlich ascites tumor

Summary Gibberellic acid in different concentrations (1000 g, 100 g, 10 g and 1 g/0.5 ml) was administered to four groups of experimental mice for nine days. Control mice received only a placebo. On the tenth day a cell suspension of Ehrlich ascites tumor (EAT) with 5×10⁶ cells in 0.1 ml was transplanted subcutaneously in the right inguinal region of all groups of mice. The animals were killed 22 days after the transplantation and the EAT growths were weighed. A comparison of the average weight of tumors grown in the experimental mice with those of the control mice showed the slowest tumor growth was in those groups of mice (p=0.001) that were pretreated with 10 and 1 g of gibberellic acid).

     

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.     

Klinisch-chemische Ultramikromethoden

Zusammenfassung Es werden Probleme und Möglichkeiten bei Arbeiten im Größenordnungsbereich von Mikrolitern (1 l=0.001 ml) dargestellt und eine Grundausrüstung beschrieben, welche es gestattet, klinisch-chemische Analysen ausgehend von kleinsten Probenmengen (5–20 l) einfach, rasch und zuverlässig auszuführen. Die Arbeitsweise von halbautomatischen Pipettenflaschen, einer Ultramikrobürette, eines speziellen Spektro-Kolorimeters und anderer apparativer Hilfsmittel wird dargestellt.

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Summary The problems and practicability of working with microliters (1 l=0.001 ml.) are presented. A basic apparatus is described, which permits the simple, rapid and reliable clinical-chemical analysis of very small samples (5–20 l.). Directions are given for the operation of semiautomatic pipettes, an ultramicroburette, a special spectrocolorimeter and other auxilliary apparatus.

     

Effects of vanadate on the c-AMP system of the heart

Summary 300 M vanadate (NH₄VO₃) caused an about 65% increase in force of contraction in isolated electrically driven cat papillary muscles. The positive inotropic effect (PIE) was accompanied by only a small (about 20%) but significant increase in c-AMP levels. 0.3 M isoprenaline (ISO) produced a much greater increase in c-AMP levels (about 80%) while the PIE of ISO was as great as the PIE observed with 300 M NH₄VO₃. In cat right ventricular adenylate cyclase preparations NH₄VO₃ and ISO stimulated adenylate cyclase activity by nearly the same extent. Propranolol (1 M and 5 M) prevented the ISO-induced adenylate cyclase stimulation but not the stimulation due to NH₄VO₃. Phosphodiesterase activity was not affected by NH₄VO₃ (up to 1000 M). It is concluded that the small vanadate-induced increase in c-AMP levels, which is due to adenylate cyclase stimulation, inintact papillary muscles maycontribute to its PIE, but other mechanisms may be at least equally important.

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Über die Wirkung von Vanadat auf das c-AMP-System des Herzens

Zusammenfassung 300 M Vanadat (NH₄VO₃) steigerte an isolierten, elektrisch gereizten Katzenpapillarmuskeln die Kontraktionskraft um ca. 65%. Der positiv inotrope Effekt (PIE) ging einher mit einem nur geringen (ca. 20%), aber signifikanten Anstieg des c-AMP-Gehaltes. 0.3 M Isoprenalin (ISO) verursachte einen sehr viel größeren Anstieg des myokardialen c-AMP-Gehaltes (ca. 80%), während der PIE von ISO gleich groß war wie der PIE von NH₄VO₃. In einer Adenylatcyclasepräparation von rechten Ventrikeln des Katzenherzens stimulierten NH₄VO₃ und ISO die Adenylatcyclaseaktivität in nahezu gleichem Ausmaß. Propranolol (1 M und 5 M) verhinderte die Aktivierung der Adenylatcyclase durch ISO, aber nicht die Aktivierung durch NH₄VO₃. Die Phosphodiesteraseaktivität wurde durch NH₄VO₃ (bis zu einer Konzentration von 1000 M) nicht beeinflußt. Aus den Ergebnissen wird geschlossen, daß der geringe vanadatinduzierte c-AMP-Anstieg, der über eine Stimulation der Adenylatcyclase zustande kommt, zwar am Zustandekommen des PIE beteiligt sein mag, daß aber andere Mechanismen von mindestens ebenso großer physiologischer Bedeutung sind.

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The data have been presented at the Symposium Cardiac Effects of Vanadate, Munich, October 26–27, 1979.

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With 1 figure and 1 table

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This work was supported by the Deutsche Forschungsgemeinschaft.     

Deleterious effects of digitalis on reperfusion-induced arrhythmias and myocardial injury in ischemic rat hearts: possible involvements of myocardial Na+ and Ca2+ imbalance

Summary Isolated rat hearts were made ischemic for 25 min after an initial recirculating perfusion, followed by 30 min of reperfusion. In some hearts, interventions including administration of ouabain and/or high [K⁺] in the buffer were performed during the first 10 min of reperfusion.During ischemia, intracellular Na⁺ (Na_i) increased from 15 to 64 [mol/g dry weight (dwt). During reperfusion, Na_i declined rapidly (at 10 min of reperfusion: 48 nol/g dwt, at 30 min: 25 mol/g dwt) and regular rhythm was recovered within 10 min in hearts without any intervention during reperfusion.⁴⁵Ca²⁺ uptake increased from 0.8 to 7.5 mol/g dwt after 30 min of reperfusion. Ventricular function recovered by 45 %.A 10-min perfusion with 10 or 50 M of ouabain increased Na_i (17 to 21 or 27 mol/g dwt) with increased left-ventricular (LV) contractile function, but these effects were reversed by combination of high perfusate [K⁺] (20 mM) in non-ischemic hearts.A 10-min reperfusion with ouabain retarded or stopped the decline in Na_i (at 10 min of reperfusion: 54 or 63 mol/g dwt, at 30 min: 32 or 40 mol/g dwt). These amounts of ouabain also increased the incidence of ventricular tachyarrhythmias during reperfusion to 30 % or 50 %, and increased the duration of ventricular fibrillation from 6.5 to 11.5 or 18.0 min.⁴⁵Ca²⁺ uptake reached to 8.8 or 10.0 mol/g dwt, and function recovered only 35 % or 28 %. When high perfusate [K⁺] was combined with ouabain during reperfusion, the retarded decline in Nai, augmented⁴⁵Ca²⁺ uptake, and reduced recovery of function caused by ouabain alone were attenuated. These results suggest that digitalis has toxic effects on reperfused ischemic hearts by inhibition of rapid active outward transport of previously elevated Na_i and potentiation of Ca²⁺ overload.The work was supported in part by grant HL 37936 from the National Heart, Lung and Blood Institute. J. R. Neely was deceased on November 29, 1988     

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.     

Serum sialic acid in malignant tumors,bacterial infections,and chronic liver diseases

Summary The total serum sialic acid concentration was determined in 2,264 persons with various malignant tumors, bacterial infections, rheumatic diseases, and chronic liver diseases, and in a control group. The thiobarbiturate method according to Warren was used [34].The upper limit (95% percentile) in the control group was 2.23 mol/ml. Higher values were found in the groups with neoplasms (mean: 3.04 mol/ml), inflammatory diseases (e.g., pneumonia: 3.02 mol/ml), and active rheumatoid arthritis (3.05 mol/ml). In the group with malignant diseases, the sialic acid concentration at the time of diagnosis was highest for bronchial carcinoma (3.29 mol/ml) and lowest for breast cancer (2.58 mol/ml). In chronic liver diseases the mean sialic acid level was lower than in a heterogeneous group of noninflammatory and nonneoplastic diseases.The estimation of the serum sialic acid concentration could be useful in the detection of tumor burden and metastases, and in the evaluation of the later course and prognosis of malignant neoplasms if bacterial/inflammatory and active rheumatoid processes can be excluded.     

C-reactive protein (CRP) levels in systemic lupus erythematosus (SLE)

Summary CRP levels in 194 serum samples from 43 SLE patients were measured. Patients with inactive disease have levels below 10 g/ml; patients with active SLE have higher levels, but never over 50 g/ml. In the presence of infection or inflammatory processes, regardless of the activity of SLE, the levels are significantly higher (p<0.05), and well over 50 g/ml. Both active SLE patients and inactive SLE patients with local infections have levels between 10 g/ml and 50 g/ml. In this situation, the presence of anti-DNA antibodies strongly suggests disease activity (82% versus 9%, p<0.05). The clinical and physiopathological meaning of these findings is discussed.     

Serum bile salts as an aid to oral cholecystogram interpretation

A radioimmunoassay for glycocholate was used as an estimate of serum bile salts in patients with serum bilirubin levels less than 5 mg/dl undergoing oral cholecystography. Most subjects also had a percutaneous liver biopsy. Intravenous cholangiography was performed in most subjects who had a nonvisualized gallbladder after receiving single doses of iopanoic acid on two consecutive days. The pathologic status of nonvisualized gallbladders was ascertained by surgery, prospective follow-up, necropsy and/or ultrasound. In 20 subjects with well-visualized gallbladders, the serum cholate conjugates (CC) were lower than 3.5 M in all but two subjects, both of whom had inflammatory liver disease (chronic active hepatitis, alcoholic hepatitis). Those with faintly or poorly visualized gallbladders, but with no evidence of gallbladder or gallstone disease by other criteria, also had levels less than 3.5 M, except for one subject with inflammatory liver disease. By contrast, 11 subjects with nonvisualized, although normal gallbladders by the above clinical criteria, exhibited serum CC greater than 3.5 M. Six subjects had diminished gallbladder visualization, but normal gallbladders on clinicopathological grounds; all but one with chronic active hepatitis had serum CC greater than 3.5 M. In conclusion, when the serum cholate conjugates are less than 3.5 M, nonvisualization of the gallbladder with oral cholecystography supports a diagnosis of a diseased gallbladder. In addition, a serum CC level greater than 3.5 M may predict insufficient hepatic capacity to secrete iopanoic acid adequate for gallbladder visualization in noninflammatory liver disease. The serum cholate conjugates appear to provide greater information on this hepatic function than the serum bilirubin or bromsulfophthalein retention.     

Kinetic and survival response of the M14 cell line to lonidamine associated with adriamycin or hyperthermia

Summary Lonidamine (LND), an indazole-carboxylic acid derivative, was delivered alone and together with adriamycin (ADM) or hyperthermia to the human melanoma cell line M14, and cell survival was assessed. Cell cycle-specific effects were investigated by analyzing sequences of DNA content histograms by means of a suitable mathematical procedure. LND delivered for 1 h at a dose of 50 g/ml did not affect proliferation and survival of the cells. Exposure of the cells for 1 h to ADM (1.0 g/ml) followed by LND for 1 h (50 g/ml) produced the highest effect on the survival. Kinetic parameters were affected by the combined treatment slightly more than by ADM exposure alone. Simultaneous delivery of LND (50 g/ml) with hyperthermia (42°C, 1 h) reduced the survival and enhanced the block of the cells in the G2M phase, as compared with the heat treatment alone. The effect of the treatments on cell survival appeared to be related to the perturbation of the G2M phase of the cycle.This research was supported by PF Oncologia CNR, grants No. 85.02422.44 and No. 102312-104348, and by AIRC, grant No. 3/85     

GlutathioneS-transferase expression in malignant mesothelioma and non-neoplastic mesothelium: an immunohistochemical study

Expression of the glutathioneS-transferase (GST) subclasses , and was investigated immunohistochemically in 20 normal or hyperplastic mesothelium and in 57 malignant mesothelioma cases. These results were correlated with survival and also with P-170 glycoprotein expression. Nearly all the non-neoplastic mesothelium cases were positive for GST and . About half of the non-neoplastic cases were positive for . Twenty-nine (51%) malignant mesotheliomas were positive for at least one of the GST species; 21 (37%) showed immunoreactivity for , 18 (31.5%) for and 21 (37%) for . A total of 54 mesothelioma cases displayed immunoreactivity for the P-170 glycoprotein. For GST and GST, a statistical significance between expression and increased survival was found (respectivelyP=0.012 and 0.024) while for GST no significance was found. The results of this study demonstrate that expression of GST correlates positively with increased survival in malignant mesothelioma. It is also concluded that, in mesothelioma, GST and P-170 glycoprotein may contribute to the resistance to cytotoxic drugs frequently observed in these tumours. No correlation between GST and P-170 expression was demonstrated.Abbreviation GST glutathioneS-transferase     

Results of a multicenter placebo-controlled double-blind randomized phase III clinical study of treatment of rheumatoid arthritis with recombinant interferon-gamma

Summary In a multicenter placebo-controlled double-blind randomized clinical study, 91 patients with rheumatoid arthritis were given 28 days' treatment with recombinant interferon-gamma (50 g daily for 20 days, then 50 g each second day up to day 28, given by subcutaneous injection). The aim of the study was to provide a methodologically clear demonstration of the efficacy of treatment with interferon-gamma, using criteria that could be handled by statistical tests. Evaluatable documentation was available for 79 patients, of whom 40 were treated with the active compound. The principal criterion for the statistical evaluation of the therapeutic success was improvement of the Ritchie joint pain index or Lansbury joint pain index by at least 30% within 28 days. The chi-square test showed superiority of the interferon arm over the placebo arm with an error probability of a<1%. In addition, efficacy of interferon-gamma was demonstrated in respect of practically all parameters investigated. The frequency of side-effects, including febrile reactions, was the same for the active compound and the placebo. During interferon treatment the daily maximum body temperature was raised by 0.3°C on average, but was below 37.2°C at all times.     

Iron stores in 1359, 30- to 60-year-old Danish women: Evaluation by serum ferritin and hemoglobin

Summary Iron status, including serum (S-)ferritin and hemoglobin (Hb), was assessed in a population survey comprising 1359 nonpregnant Danish women in age cohorts of 30, 40, 50, and 60 years. S-ferritin levels were similar in 30- and 40-year-old women; they displayed a significant increase in 50-year-old women and a further significant increase in 60-year-old women. In the 30- and 40-year-old women, median S-ferritin was 38g/l, 5–95 percentile 6–135g/l; 17.2% had values < 15,g/l (i.e., depleted iron stores), 22.7% values from 15 to 30g/l (i.e., small iron stores), and 60.1% values > 30g/l (i.e., replete iron stores). In the 50-year-old women, median S-ferritin was 54g/l, 5–95 percentile 10–164g/l; 10.3% had values < 15g/l, 16.5% values from 15 to 30g/l, and 73.2% values > 30g/l. For the 60-year-old women, median S-ferritin was 84g/l, 5–95 percentile 25–249g/l; 1.6% had values < 15g/l, 8.6% values from 15 to 30g/l, and 89.8% values > 30g/l. Blood donors (n=180) had lower S-ferritin than nondonors in all age-groups (p<0.001). In the entire series, Hb levels were similar in 30- and 40-year-old women, median 137 g/l (8.5 mmol/l), 5–95 percentile 121–152 g/1 (7.5–9.4 mmol/l), and higher in 50- and 60-year-old women, median 140 g/l (8.7 mmol/l), 5–95 percentile 123-158 g/l (7.6–9.8 mmol/l) (p<0.0001). Hb values < 121 g/l (7.5 mmol/l) were observed in 3.8% of the women. Women with S-ferritin < 15 g/l (n=161) had lower Hb, median 134 g/l (8.3 mmol/l), than those with S-ferritin > 15 g/l, median 139 g/l (8.6 mmol/l) (p<0.001). Iron deficiency anemia (S-ferritin < 15 g/l and Hb < 121 g/l) was seen in 2.3% of 30- and 40-year-old women, and in 1.1% of 50- and 60-year-old women.     

Coronary heart disease and urinary albumin excretion rate in Type 2 (non-insulin-dependent) diabetic patients

Summary Associations between overnight urinary albumin excretion rate and prevalent coronary heart disease and its major risk factors were examined in a cross-sectional study of 141 Type 2 (non-insulin-dependent) diabetic patients. Mean albumin excretion rate was higher in men (geometric mean 13.5 g/min; 95% confidence interval 10.3–17.6) than women (7.5 g/min; 5.7–9.8, p<0.01). In diabetic men and women mean albumin excretion rate was higher in those with electrocardiographic and/or symptomatic evidence of coronary heart disease than in those without (men, 23.1 g/ min; 95% confidence interval 13.7–39.0 versus 10.6 g/min; 7.9–14.2, p<0.01, women, 13.7 g/min; 8.0–23.5 versus 5.4 g/min; 4.2–6.8, p<0.01). Multiple logistic regression analysis was used to allow for confounding between variables. In the diabetic group as a whole, raised albumin excretion rate (p<0.001), gender (p<0.05) and systolic blood pressure (p=0.06) entered the best model for coronary heart disease prediction. In women, albumin excretion rate alone (p<0.01) and in men albumin excretion rate (p<0.01) and age (p=0.05) entered the best models. We conclude that albumin excretion rate is significantly associated with coronary heart disease morbidity after taking into account the confounding effects of raised blood pressure and other cardiovascular risk factors.     

Neural influences on human esophageal and salivary alkali secretion

Esophageal secretion of HCO ₃ ^– ions occurs in opossum and man and may contribute to mucosal defense. Using a perfusion technique, neuroregulatory influences on esophageal and salivary HCO ₃ ^– secretion were investigated in 24 healthy human subjects. The sight and smell of food increased median salivary HCO ₃ ^– output from 424 to 573 mol/15 min (P=0.014), without significantly altering esophageal HCO ₃ ^– secretion (74–105 mol/15 min,P=0.24). Atropine reduced both salivary (610 to 68, 17, 10, and 3 mol/15 min in successive periods;P<0.028) and esophageal HCO ₃ ^– output (108 to 78, 35, 18, and 7 mol/10 cm/15 min;P<0.028), respectively. Following atropinization, cholinergic stimulation failed to increase salivary secretion but did unmask a small rise in esophageal alkali output (7 to 27 mol/10 cm/15 min,P=0.036), implicating a noncholinergic mechanism. Cold-induced pain activated sympathetic reflexes and reduced esophageal HCO ₃ ^– output (91 to 64 mol/10 cm/15 min,P=0.041) without influencing salivary secretion. These observations support a role for the autonomic nervous system in modulating human esophageal and salivary HCO ₃ ^– secretion.     

Influence of phorbol esters on contractile force,action potential and calcium current of isolated guinea-pig heart tissues

Phorbol-12,13-dibutyrate (PDB) reduced concentration-dependently the contractile force of guineapig papillary muscles (EC₅₀ 1.07 mol/l) while phorbol-12-myristate-13-acetate (PMA) was ineffective. The protein kinase C inhibitors staurosporine (0.1 mol/l) and polymyxin B (70 mol/l) did not antagonize the negative inotropic effect of PDB. Neither PMA nor PDB, in concentrations up to 30 mol/l caused significant changes of the membrane resting potential, the maximum depolarization velocity, the action potential duration or the functional refractory period in intact papillary muscles. In isolated ventricular cardiomyocytes the inward calcium current was halved by either 1 mol/l PDB or 10 mol/l PMA. PKC inhibitors attenuated, but could not completely abolish this effect of the phorboles. It is concluded that the negative inotropic effect of PDB is caused by a reduction of the slow inward calcium current and that this inhibition is, for the greater part, not mediated by an activation of protein kinase C.     

Synovial nitric oxide concentrations are increased and correlated with serum levels in patients with active Behçet’s disease: a pilot study

Behçets disease (BD) is a relapsing immunoinflammatory vasculitis of unknown etiology characterized by endothelial dysfunction. Articular symptoms and signs are present in about 75% of cases and characterized by seronegative arthritis and nonspecific synovitis. We demonstrated that both serum and erythrocyte nitric oxide (NO^·) levels, the most abundant free radical in the body, were elevated in BD and associated with disease activity. This study further investigated NO^· levels in the synovial fluid and serum from patients with active and inactive BD. A total of 23 BD patients with articular involvement (14 men and 9 women) satisfying International Study Group criteria and 15 age- and sex-matched healthy control subjects (9 men and 6 women) undergoing elective arthroscopy were included in this case-control investigation. The synovial fluid and serum were obtained from BD patients and controls. Clinical and laboratory findings including neutrophil count and erythrocyte sedimentation rate (ESR) were used to classify BD patients as active (n=11) or inactive (n=12). Synovial as well as serum NO^· levels were compared between the groups and correlation analysis was performed. Acute phase reactant levels were significantly higher (for each, p<0.01) in BD patients than control subjects in the active period. The mean synovial NO^· level in active Behçets patients (mean±SD 76.61±11.95 mol/l) was significantly higher than in inactive patients (46.16±8.89 mol/l, p<0.001) and healthy control subjects (39.60±8.03 mol/l, p<0.001). The difference between inactive patients and controls was not significant (p>0.05). Active BD patients had significantly higher serum NO^· levels (38.84±9.15 mol/l) than inactive patients (30.91±5.88 mol/l, p=0.018) and control subjects (28.86±5.91 mol/l, p=0.002). In addition, synovial NO^· levels were positively correlated with serum levels (r²=0.621, p<0.001). Increased synovial NO^· levels in active BD patients probably reflect a nonspecific inflammatory process of the synovium and, therefore, arthralgia and arthritis as a common finding of BD.