Liver microsomal Δ6 and Δ5 linoleic acid desaturation in female BB rats

Summary Rat liver microsomal 6 and 5 desaturation are defective in experimental diabetes, but this defect is correctable with insulin treatment. Rat liver fatty acid composition and 6 and 5 desaturation were studied in the spontaneously diabetic adult female Bio-Breeding (BB) rat. Control Wistar rats and BB rats (4 weeks of diabetes), that received insulin (1 IU·100 g body weight^–1·day^–1), were killed 20 h after the last insulin injection. 6 and 5 desaturase activities were estimated from the incubation of liver microsomes with (1-¹⁴C) 18:2, n–6 or (2-¹⁴C) 20:3, n–6, respectively, and the fatty acid composition of the liver and microsomal liver lipids were investigated. Under experimental conditions 6 and 5 desaturase activities were unchanged in the BB rats when compared to the control rats. Impairment of the liver fatty acid composition of diabetic BB rats is not consistent with normal desaturase activity and may be explained by factors other than desaturation disturbance.     

Insulin-like growth factor-I in man enhances lipid mobilization and oxidation induced by a growth hormone pulse

Summary Growth hormone (GH) secretion is suppressed during insulin-like growth factor-I (IGF-I) administration. The aim of the study was to examine whether IGF-I alters the metabolic response to a GH pulse. Seven healthy male subjects (age 27±4 years, BMI 21.8±1.7 kg/m²) were treated with NaCl 0.9% (saline) or IGF-I (8 g · kg^–1 · h^–1) for 5 days by continuous subcutaneous infusion in a randomized, crossover fashion while receiving an isocaloric diet (30 kcal · kg^–1 · day^–1). On the third treatment day an intravenous bolus of 0.5 U GH was administered. Forearm muscle metabolism was examined by measuring arterialized and deep venous blood samples, forearm blood flow by occlusion plethysmography and substrate oxidation by indirect calorimetry. IGF-I treatment significantly reduced insulin concentrations by 80% (p<0.02) and C-peptide levels by 78% (p<0.02), as assessed by area under the curve. Non-esterified fatty acid (NEFA), glycerol and 3-OH-butyrate levels were elevated and alanine concentration decreased. Forearm blood flow rose from 2.10±0.43 (saline) to 2.79±0.37 ml · 100ml^–1 · min^–1 (IGF-I) (p<0.02). GH-pulse: 10 h after i.v. GH injection serum GH peaked at 40.9±7.4 ng/ml. GH did not influence circulating levels of total IGFI, C-peptide, insulin or glucose, but caused a further increase in NEFA, glycerol and 3-OH-butyrate levels, indicating enhanced lipolysis and ketogenesis. This effect of GH was much more pronounced during IGF-I: NEFA rose from 702±267 (saline) and 885±236 (IGF-I) to 963±215 (saline) (p<0.05) and 1815±586 mol/l (IGF-I) (p<0.02), respectively; after 5 h, 3-OH-butyrate rose from 242±234 (saline) and 340±280 (IGF-I) to 678±638 (saline) (p<0.02) and 1115±578 mol/l (IGF-I) (p<0.02) respectively. After injection of GH, forearm uptake of 3-OH-butyrate was markedly elevated only in the subjects treated with IGF-I: from 44±195 to 300±370 after 20 min (p<0.03) and to 287±91 nmol · 100 ml^–1 · min^–1after 120 min (p<0.02). In conclusion, the lipolytic and ketogenic response to GH was grossly enhanced during IGF-I treatment, and utilization of ketone bodies by skeletal muscle was increased.Abbreviations AUC Area under the curve - C-peptide connecting peptide - EE energy expenditure - FFM fat-free mass - GH growth hormone - IGF-I insulin-like growth factor-I - NEFA non-esterified fatty acid - Ra rate of glucose appearance - Rd rate of glucose disposal - FGU forearm glucose uptake - CV coefficient of variation     

Clinical application of wave intensity for the treatment of essential hypertension

Wave intensity analysis is a method of studying intravascular flow wave propagation, calculated as the product of the rate of change in pressure (dP/t) and the rate of change in velocity (dU/dt). The typical pattern of wave intensity seen during the cardiac cycle has two dominant peaks. The larger first peak (FP) occurs during early systole when a forward traveling compression wave is generated by the left ventricle. The second smaller peak (SP) follows a period of relatively little net wave production during mid-systole. Wave reflection is seen as a small backward-traveling compression wave occurring just after the first peak of wave intensity (NP). In this study, we investigated the usefulness of parameters from the wave intensity for estimating the efficacy of the -1 blocker, doxazosin, to reduce blood pressure, by the reduction of peripheral vascular resistance. We examined 20 patients with essential hypertension. Patients were included if their diastolic blood pressure was 95mmHg on at least three separate visits to the clinic. The study consisted of a 2-week baseline phase followed by a 2–4-week dose-adjusted phase with doxazosin. Treatment began with 1mg/day doxazosin, and the dose was doubled fortnightly until the diastolic blood pressure was 90mmHg. Blood–pressure measurements and side effects were recorded at intervals of 2 weeks. Before and after 4 weeks of stable treatment with doxazosin, a comprehensive clinical evaluation was given. Doxazosin reduced systolic and diastolic blood pressure. Both FP and SP increased and NP decreased. MBP (change in mean blood pressure) correlated well with NP before and after the antihypertensive therapy. The efficacy of doxazosin was confirmed by the decreased reflection wave of aortic flow from wave intensity analysis. Thus, patients with a significant reflection wave may be good candidates for antihypertensive treatment by a vasodilator, such as doxazosin.     

α-Adrenergic responses of isolated canine coronary microvessels

Although -adrenergic activation is known to increase coronary microvascular resistance in vivo, the magnitude of its segmental microvascular consequences is not well understood. Quantification of these effects in vivo is hindered by escape mechanisms that minimize the influences of constrictors, and alterations in flow and pressure, which effect microvascular tone by shear stress-dependent and myogenic mechanisms, respectively. To eliminate these confounding influences, we have studied responses in vitro under conditions with these variables controlled. We evaluated the diameter changes of isolated canine coronary arterioles (110±12 m, n=35) and venules (98±7 m, n=9) in response to -adrenergic activation by norepinephrine (10^–10 to 10^–4 M) in the presence of -adrenergic blockade by alprenolol (10^–6 M). In contrast to the situation in vivo, -adrenergic activation did not constrict isolated coronary arterioles, but constricted isolated coronary venules in a dose-dependent manner over a range of 10^–10 to 10^–4 M (–27 ±3% maximum diameter change). Coronary arteriolar -adrenergic constriction was not promoted by 1) subthreshold or vasoactive doses of the vasoconstrictors KCl, angiotensin II, U46619, endothelin-1, neuropeptide Y or arginine vasopressin, 2) inhibition of the presynaptic uptake of norepinephrine by imipramine (10^–6 M), 3) inhibition of EDRF synthesis by N^g-monomethyl-L-arginine (10^–5 M) or 4) inhibition of prostaglandin synthesis by indomethacin (10^–5 M). Furthermore, -adrenergic activation did not modify microvascular dilatation by adenosine (10^–9 to 10^–4 M) or nitroglycerin (10^–9 to 10^–4 M), suggesting that -adrenergic constriction in vivo is not due to attenuation of cAMP or cGMP-dependent mechanisms of coronary dilatation. In contrast to the lack of constriction in coronary arterioles, canine skeletal muscle arterioles exhibited significant -adrenergic constriction (–80±4%), maximum diameter change). The coronary venular -adrenergic constriction was significantly inhibited by both the ₁-and ₂-adrenergic receptor antagonists, prazosin (10^–8 M) and rauwolscine (10^–7 M), indicating a mixed population of ₁-and ₂-adrenergic receptors. These results suggest that coronary arterioles, but not venules, lose -adrenergic responsiveness during isolation and cannulation, or that the primary coronary microvascular response to -adrenergic activation is venular constriction.     

Association of systemic and thyroid autoimmune diseases

Objective: There are few large cohort studies available on the association of systemic and thyroid autoimmune diseases. In this study, we wished to determine the association of Hashimotos thyroiditis (HT) and Graves disease (GD) with systemic autoimmune diseases. Methods: One thousand five hundred and seventeen patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), mixed connective tissue disease (MCTD), Sjögrens syndrome (SS) and polymyositis/dermatomyositis (PM/DM) were included in the study. The HT and GD were diagnosed based on thorough clinical evaluation, imaging and fine-needle aspiration cytology (FNAC). The frequency of HT and GD in these diseases was assessed. In addition, 426 patients with HT or GD were assessed and the incidence of SLE, RA, SSc, MCTD, SS and PM/DM among these patients was determined. Prevalence ratios indicating the prevalences of GD or HT among our autoimmune patients in comparison to prevalences of GD or HT in the general population were calculated. Results: Altogether 8.2% of systemic autoimmune patients had either HT or GD. MCTD and SS most frequently overlapped with autoimmune thyroid diseases (24 and 10%, respectively). HT was more common among MCTD, SS and RA patients (21, 7 and 6%, respectively) than GD (2.5, 3 and 1.6%, respectively). The prevalences of HT in SLE, RA, SSc, MCTD, SS and PM/DM were 90–, 160–, 220–, 556–, 176– and 69-fold higher than in the general population, respectively. The prevalences of GD in the same systemic diseases were 68-, 50-, 102-, 76-, 74- and 37-fold higher than in the general population, respectively. Among all thyroid patients, 30% had associated systemic disease. In particular, 51% of HT and only 16% of GD subjects had any of the systemic disorders. MCTD, SS, SLE, RA, SSc and PM/DM were all more common among HT patients (20, 17, 7, 4, 2 and 2%, respectively) than in GD individuals (2, 5, 5, 1, 2 and 1%, respectively). Conclusion: Systemic and thyroid autoimmune diseases often overlap with each other. HT and GD may be most common among MCTD, SSc and SS patients. On the other hand, these systemic diseases are often present in HT subjects. Therefore it is clinically important to screen patients with systemic autoimmune diseases for the co-existence of thyroid disorders.Take home message: Autoimmune thyroid diseases, such as Hashimotos thyroiditis and Graves disease are often associated with systemic autoimmune diseases, most commonly with Sjögrens syndrome and mixed connective tissue disease.     

Low acute hematological toxicity during chemotherapy predicts reduced disease control in advanced Hodgkin’s disease

Chemotherapy-treated patients with advanced Hodgkins disease (HD) differ considerably in acute hematotoxicity. Hematotoxicity may be indicative of pharmacological and metabolic heterogeneity. We hypothesized that low hematotoxicity might correlate with reduced systemic dose and thus reduced disease control. A total of 266 patients with advanced HD treated with cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, vinblastine, and dacarbazine (COPP-ABVD) were analyzed (HD6 trial of the German Hodgkins Lymphoma Study Group). The reported WHO grade of leukocytopenia was averaged over chemotherapy cycles given and weighted with the reciprocal dose intensity of the corresponding cycle. The low and high toxicity groups were defined in retrospect as having had an averaged WHO grade of leukocytopenia 2.1 and >2.1, respectively. The independent impact of low hematological toxicity on freedom from treatment failure (FFTF) was assessed multivariately adjusting for the international prognostic score for advanced HD. The results were validated in two independent cohorts [181 patients treated with COPP-ABVD (HD9-trial) and 250 patients treated with COPP-ABV-ifosfamide, methotrexate, etoposide, and prednisone (IMEP) (HD6 trial)]. The 5-year FFTF rates were 68% for patients with high toxicity vs 47% for patients with low toxicity [multivariate relative risk (RR) 2.0, 95% confidence interval (CI) 1.4–3.0, p=0.0002]. Patients with low toxicity received significantly higher nominal dose (p=0.02) and dose intensity (p<0.0001). This finding was confirmed in both validation cohorts (multivariate RR 2.1, 95% CI 1.2–3.8, p=0.01 and RR 1.5, 95% CI 1.01–2.26, p=0.04, respectively). Patients with low hematotoxicity have significantly higher failure rates despite higher doses and dose intensity. Hematotoxicity is an independent prognostic factor for treatment outcome. This observation suggests a strategy of individualized dosing adapted to hematotoxicity.The authors listed above wrote this contribution on behalf of the German Hodgkins Lymphoma Study Group     

Effect of starvation upon insulin secretion in the rabbit

Summary Plasma insulin and blood glucose levels of normal and fasting rabbits have been studied. The mean plasma insulin of pre-starvation controls was 24 µU/ml; blood glucose was 107 mg/100 ml. After starvation, the insulin levels in the blood decreased to 8 µU/ml and glucose concentration to a mean level of 76 mg/100 ml. The fine structure of - and -cells was studied in fasted animals. Most of the -cells were filled with granules of the pale type and some of these secretory granules were adjacent to the plasmatic membrane, which frequently showed projecting microvilli. Immunoassay figures for pancreatic insulin were obtained. There was a good correlation between the insulin assay results in pancreas on one hand, and light and electron microscopic -cells on the other, provided the assumption is true that pale granules contain immunologically active insulin. Most of the -cells were well granulated, but a few of these cells showed a marked increase in the lamellar ergastoplasm.This work has been supported by a grant from the Seguridad Social del Estado, Madrid.     

Luminal dopamine modulates canine ileal water and electrolyte transport

Previous studies have suggested that dopamine stimulates active ileal ion absorption via ₂-adrenergic or dopaminergic receptor activation. Identification of a dopamine 1a receptor on rat enterocytes located in intestinal crypts prompted this investigation of the effect of luminally administered dopamine on water and ion transport in the canine ileum. Absorption studies (n=27) were performed in dogs with 25-cm ileal Thiry-Vella fistulas. Perfusion with [¹⁴C PEG was used to calculate absorption of water and electrolytes from the Thiry-Vella fistula. Experiments consisted of three 1-hr periods: basal, luminal drug infusion at 10^–4 M, and recovery. Agonists used included dopamine (DOP: -adrenergic, D₁ and D₂ receptor) and SKF 38393 (D₁ receptor). Antagonists used included terazosin (TZ: ₁) and yohimbine (YOH: ₂). DOP caused significant increases in water and electrolyte absorption. TZ and YOH prevented the dopamine-induced proabsorptive response. Luminal DOP may serve as a proabsorptive modulator of ileal transport, acting via ₁, ₂, and dopaminergic receptors. The development of more potent proabsorptive dopamine analogs, which maintain the ability to broadly activate mucosal receptors, may be useful in such clinical situations as diabetic diarrhea, short gut syndrome, or following small bowel transplantation.Presented in part as a poster presentation at the Annual Meeting of the American Gastroenterological Association, New Orleans, May 14–18, 1994, and published in abstract form inGastroenterology 106:A432, 1994.Supported in part by National Institutes of Health grant R29-DK 41178 (C.J.Y.).     

Effect of α2-adrenoceptor antagonist on platelet activation during insulin-induced hypoglycaemia in Type 2 (non-insulin-dependent) diabetes mellitus

Summary The role of epinephrine in platelet activation and the effect of an ₂-adrenoceptor antagonist, midaglizole, during insulin-induced hypoglycaemia in Type 2 (noninsulin-dependent) diabetes mellitus were examined. The action of midaglizole as a platelet ₂-antagonist was confirmed by in vitro studies using platelet-rich plasma and washed platelet suspension. Hypoglycaemia was induced by a bolus injection of short-acting insulin in 24 diabetic patients. They were divided into two groups, a control group (n=12) and an ₂-group (n=12), and midaglizole was administered orally 60 min before insulin injection in the latter. Blood glucose and plasma C-peptide levels were significantly decreased (p<0.005) by insulin injection in both groups. Counter-regulatory hormones, including epinephrine, and arginine vasopressin were similarly increased at the hypoglycaemic nadir compared with the levels at 0 min in both groups. Plasma -thromboglobulin was increased at the hypoglycaemic nadir (165.5±12.6ng/ml) compared with the level at 0 min (121.0±11.5, p<0.005) in the control group, whereas no significant increase was demonstrated in the ₂ group. These results suggest that plasma epinephrine plays an important role in platelet activation during hypoglycaemia in Type 2 diabetes mellitus, and that the platelet activation is prevented by ₂-adrenoceptor antagonist.     

Differences of cellular composition and adhesion molecule expression in “leukemic” as compared with “normal” human long-term bone marrow cultures

Summary Human long-term bone marrow cultures (HLTBMCs) were established with bone marrow samples collected from 15 patients with acute myeloid leukemia (AML) and compared with HLTBMCs from eight healthy volunteers. During 6 weeks of culture, the cellular composition of HLTBMCs was quantitatively studied. The cells of the HLTBMCs were divided into three main categories: fibroblasts, macrophages, and other cells (endothelial cells, hematopoietic cells and undefined cells). HLTBMCs derived from healthy volunteers demonstrated a very consistent development. The number of fibroblasts increased during culture and the number of macrophages decreased, resulting in a steady state after 3 weeks of culture. In contrast, HLTBMCs derived from patients with AML showed a strikingly different pattern of irregular development and a steady state was not reached under our conditions. The APAAP technique was used to demonstrate expression of adhesion molecules. VLA2, VLA5, VLA6, LFA1, Mac1, p150/95, ₂-chain, HCAM, ICAM1, NCAM, and VCAM1 were more expressed on normal as compared with leukemic bone marrow stromal cells, although this reached significance only for ₂-chain and NCAM. VLA1, 3, and 4 were expressed in a higher percentage on leukemic stroma (not significant). More expression was seen on normal as opposed to leukemic macrophages for the adhesion molecules tested, except for VLA5. The differences reached significance for the majority of molecules tested. It is concluded that striking differences exist in cellular composition and adhesion molecule expression between HLTBMCs from healthy individuals and those from patients with AML. This may have an impact on the pathogenesis of AML.     

Delorme’s Procedure for Rectal Prolapse

PURPOSE: Clinical and physiological results of Delormes procedure were assessed retrospectively in patients undergoing this procedure for rectal prolapse. METHODS: A consecutive series of 31 patients (7 males, 24 females; age, 14–93, mean 70 years) with full-thickness, rectal prolapse were treated by Delormes procedure between 1994 and 2002. Median follow-up was 39 (range, 6–96) months. RESULTS: Good results were achieved in 27 patients (87 percent), prolapse recurrence was observed in 4 (13 percent), and mean recurrence time was 14 (range, 3–25) months. There were no postoperative deaths. Minor complications occurred in four patients. The median changes in preoperative and postoperative physiologic patterns in 16 patients were as follows: resting pressure from 21.0 (range, 5–48) to 23.5 (range, 12–76) cm H2O (P = 0.030), squeeze pressure from 64.0 (range, 27–248) to 108.0 (range, 32–264) cm H2O (P = 0.041), volume at first sensation from 100 (range, 70–180) to 70 (range, 40–130) ml (P = 0.002), maximum tolerated volume from 260 (range, 120–400) to 160 (range, 70–400) ml (P = 0.001). Incontinence improved in 63 percent. No patient became constipated, and 38 percent of those constipated preoperatively improved. The preoperative incontinence score improved from 11.5 (range, 1–20) to 6.0 (range, 0–20) after operation (P < 0.0001). CONCLUSION: Delormes procedure had a low morbidity, did not lead to constipation, improved anal continence, and had a reasonably low recurrence rate. Improved anal sphincter and rectal sensation were associated with a reduced incidence of defecatory problems after Delormes procedure.     

Thalassemia intermedia: compound heterozygous β∘/β+-thalassemia and co-inherited heterozygous α+-thalassemia

Summary The relative excess of - over -globin chains in the erythroid precursors is the chief pathophysiological factor of homozygous -thalassemia. The clinical picture is usually characterized by a transfusion-dependent dyserythropoietic anemia (thalassemia major). However, some patients present with moderate anemia that does not require regular blood transfusions (thalassemia intermedia). The molecular heterogeneity of -thalassemia mutations and changes of - and -globin gene expression play an important role in modifying the clinical phenotype. We report here on a female Greek patient with homozygous -thalassemia but normal growth and development, excellent exercise tolerance, and no need of blood transfusions. She is thus mildly affected clinically, although there is marked pallor, jaundice, and hepatosplenomegaly. These signs correspond to her marked hypochromic, microcytic anemia with erythroid hyperplasia of the bone marrow. -Globin genotyping shows her to be compound heterozygous for the codon 39 C T -nonsense mutation and for the T C ⁺-mutation at position 6 of the splice consensus at the exon 1/intron 1 junction (CD39 C T/IVS 1–6 T C). -Globin gene mapping demonstrates the presence of a 3.7-kb ⁺-thalassemia deletion on one allele (–^3.7/). Taken together, this study identifies a complex interaction of genetic factors that do not significantly alter the clinical phenotype when present alone but ameliorate the course of homozygous -thalassemia when inherited in combination.Abbreviations Hb hemoglobin - Hct hematocrit - HPFH hereditary persistence of fetal hemoglobin - IVS intervening sequence - MCH mean corpuscular hemoglobin - MCV mean corpuscular volume - PCR polymerase chain reaction     

Effects of glucocorticoids and insulin on 3′,5′-AMP phosphodiesterase activity in adrenalectomized rats

Summary Glucocorticoids stimulate the rate of lipolysis which is reduced in adrenalectomized animals. This hormonal action is antagonized by insulin. The antilipolytic action of insulin appears to be mediated by a reduced intracellular concentration of 3,5-AMP. This reduction can partly be attributed to an insulin-induced acceleration of 3,5-AMP degradation. — It is shown that the stimulatory influence of glucocorticoids on lipolysis is due to a reduction of 3,5-AMP phosphodiesterase (PDE) activity, which is increased by adrenalectomy. PDE activity was also increased in liver, skeletal muscle and kidney of adrenalectomized rats; treatment with a glucoeorticoid prevented this increase.In vitro, PDE purified from beef heart was inhibited by glucocorticoids in high concentrations (K_i=1.1 · 10^–3 M for 6 -methylprednisolone hemisuccinate,K _i=1.6 · 10^–3 M for prednisolone succinate).In vivo, the glucocorticoid-induced decrease of PDE activity (with retarded onset as shown in liver), may essentially be attributed to a decreased enzyme synthesis. — Studies on the interaction of insulin and glucocorticoids on PDE activity were performed in the liver. In adrenalectomized, alloxan diabetic rats insulin stimulated PDE activity suppressed by treatment with a glucoeorticoid, unsuppressed PDE activity was not increased by insulin. In contrast, the action of glucocorticoids on PDE activity was independent of the presence or the effectiveness of insulin.

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Wirkungen von Olucocorticoiden und Insulin auf die 3,5-AMP-Phosphodiesterase-Aktivität bei adrenalektomierten Tieren

Zusammenfassung Die Lipolyse, die bei adrenalektomierten Tieren vermindert ist, wird durch Glucocorticoide verstärkt. Die gegenüber Glucocorticoiden antagonistische Wirkung von Insulin, das die Lipolyse vermindert, kann durch eine Abnahme der intracellulären 3,5-AMP-Konzentration erklärt werden. Diese ist teilweise auf einen beschleunigten Abbau des Nucleotids zurückzuführen. — Die Lipolysesteigerung durch Glucocorticoide ist durch eine Verminderung der 3,5-AMP-Phosphodiesterase (PDE)-Aktivität bedingt, die bei adrenalektomierten Ratten erhöht ist. Die PDE-Aktivität adrenalektomierter Tiere ist auch in Leber, Skeletmuskulatur und Niere erhöht, die Gabe eines Glucocorticoids verhindert diesen Anstieg. Glucocorticoide hemmen aus Rinderherz isolierte PDEin vitro, jedoch sind hohe Steroidkonzentrationen erforderlich (K _i=1.1 · 10^–3 M für 6 -Methylprednisolon-Hemisuceinat, (K _i=1,6 · 10^–3 M für Prednisolon-Succinat). Die einige Stunden nach Gabe eines Glucocorticoids einsetzende Abnahme der PDE-Aktivität kann im wesentlichen auf eine verminderte Enzymsynthese zurückgeführt werden. — Insulin steigert bei adrenalektomierten, alloxandiabetischen Ratten die PDE-Aktivität in der Leber nur, wenn die Tiere mit einem Glucocorticoid behandelt sind, nicht jedoch die erhöhte PDE-Aktivität bei Fehlen von Glucocorticoiden. Der Einfluß von Glucocorticoiden auf die PDE-Aktivität ist dagegen nicht an die Wirkung von Insulin gebunden.

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Effets des glucocorticoïdes et de l'insuline sur l'activité de la 35-AMP-phosphodiestérase chez des rats surrénalectomisés

Résumé Les glucocorticoïdes stimulent la lipolyse qui est réduite chez les animaux surrénalectomisés. Cette action hormonale est contrecarrée par l'insuline. L'action antilipolytique de l'insuline semble être due à une réduction de la concentration intracellulaire de 35-AMP. Cette réduction peut être attribuée en partie à une accélération due à l'insuline de la dégradation du 35-AMP. On a montré que l'influence stimulatrice des glucocorticoïdes sur la lipolyse est due à une réduction de l'activité de la 35-AMP-phosphodiesterase (PDE), qui est accrue par la surrénalectomie. L'activité de la phosphodiesterase est également augmentée dans le foie, le muscle strié et les reins des rats surrénalectomisés, le traitement par un glucocorticoïde prévient cette augmentation.In vitro, la phosphodiesterase purifiée du coeur de boeuf est inhibée par les glucocorticoïdes à fortes concentrations (K _i=1.1 · 10^–3 M pour l'hémisuccinate de 6 -méthylprednisolone, (K _i=1.6 · 10^–3 M pour le succinate de prednisolone).In vivo, la diminution provoquée par les glucocorticoïdes de l'activité de la phosphodiesterase qui se produit au bout de quelque temps, comme on l'a montré dans le foie, peut essentiellement être attribuée à une synthèse diminuée de l'enzyme. Des études sur l'interaction de l'insuline et des glucocorticoïdes sur l'activité de la phosphodiesterase ont été effectuées sur le foie. Chez les rats surrénalectomisés, rendus diabétiques par l'alloxane, l'insuline stimule l'activité de la phosphodiesterase qui a été supprimée par un traitement avec un glucocorticoïde, l'activité de la phosphodiesterase qui n'a pas été supprimée n'est pas augmentée par l'insuline. Par contre, l'action des glucocorticoïdes sur l'activité de la phosphodiesterase est indépendante de l'action de l'insuline.

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Non-Standard Abbreviations G 6 P Glucose-6-phosphate - FFA non-esterified, free fatty acids - 3,5-AMP cyclic adenosine-3,5-monophosphate - PDE 3,5-AMP phosphodiesterase This study was supported by the Deutsche Forschungsgemeinschaft.Deceased October 31, 1967.     

Overproduction of inhibitory hematopoietic cytokines by lipopolysaccharide-activated peripheral blood mononuclear cells in patients with aplastic anemia

The aim of this study was to measure the level of cytokines produced by peripheral blood mononuclear cells (PBMNC) in patients with aplastic anemia (AA) and to determine their effect on the clonal growth of normal bone marrow (BM) cells. Twenty-one patients with AA and 11 normal controls were enrolled in this study. Medium conditioned by PBMNC of AA patients in the presence of lipopolysaccharide (LPS) was found to be suppressive to the colony growth of normal BM cells. Thus, we further determined the presence in the PBMNC-conditioned medium (CM) of both inhibitory cytokines: macrophage inflammatory protein-1 (MIP-1), tumor necrosis factor- (TNF-), transforming growth factor-2 (TGF-2), and interferon- (IFN-), and stimulatory cytokines: interleukin-3 (IL-3) and stem cell factor (SCF). Spontaneous production of MlP-1 was higher in the AA patients than the normal controls (1887±174 pg/ml vs 1643±93 pg/ml), but the difference was not significant. After LPS stimulation, the production of MIP-1 was markedly increased in the AA patients, and its level was significantly higher than that of the normal controls (2360±149 pg/ml vs 1517±92 pg/ml, p=0.0022). The level of TNF was also higher in the AA patients. However, IFN-, TGF-2, SCF, and IL-3 were not detectable in the PBMNC-CM of either AA patients or normals. The myelopoietic suppressing effect of AA-PBMNC-CM from each AA patient was significantly blocked by pretreatment with anti-TNF-, resulting in a colony-forming enhancement of 174%±12%. A similar effect was noted in six of 11 AA patients by pretreatment with anti-MIP-1. We conclude that TNF and MIP-1 can be overproduced by the PBMNC of some AA patients, which may play a role in the progression of AA.     

Biliary Interleukin-6 and Tumor Necrosis Factor-α in Patients Undergoing Endoscopic Retrograde Cholangiopancreatography

Cytokines are low-molecular-weight proteinmediators that possess a wide spectrum of inflammatory,metabolic, and immunomodulatory properties. Cytokineshave been shown to be produced by monocytes/macrophages, lymphocytes, fibroblasts, endothelial cells,and more recently, hepatocytes and biliary epithelium.The aim of this study was to define biliary levels ofinterleukin-6 (IL-6) and tumor necrosis factor- (TNF-) in patients undergoing endoscopicretrograde cholangiopancreatography (ERCP) in variousdisease states. Fifty-four patients undergoing ERCPcomprised the study group. IL-6 and TNF- were measured in aspirated bile using an ELISAtechnique. Levels of both TNF- and IL-6 weresignificantly higher in patients with cholangitis (P< 0.00001). Moreover, IL-6 was 100% specific forcholangitis since none of the patients without bacterialcholangitis — including patients with biliaryobstruction secondary to cholangiocarcinoma orpancreatic carcinoma — had measurable IL-6 intheir bile. Low levels of biliary TNF- were detectable in fivepatients without cholangitis; the sensitivity andspecificity of TNF- for cholangitis were 100% and82%, respectively. There was a strong statisticalcorrelation between biliary IL-6 and TNF- levels (r= 0.819, P < 0.0001). In contrast, the correlationsbetween biliary cytokines and serum biochemicalparameters were weak. These results suggest that IL-6and TNF- are sensitive markers for cholangitis and maydifferentiate it from other types of biliary tractdisease.     

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.     

Involvement of Luminal Bacteria, Heat Shock Protein 60, Macrophages and γδ T Cells in Dextran Sulfate Sodium-Induced Colitis in Rats

The in vivo immunological events in dextran sulfate sodium (DSS) -induced colitis were evaluated. Rats were fed water (control) or 5% DSS. Colonic sections were assessed by light microscopy, Gram stain, immunohistochemistry, and electron microscopy. A progressive decline in number and increase in fragmentation of bacteria in the colonic lumen was observed over time. Luminal bacteria were the first to show heat shock protein 60 (HSP60) staining (day 3). Macrophages in close proximity to these bacteria were next to show such staining (day 6), and finally the damaged epithelial cells when colitis became severe (day 15). Ultrastructural assessment showed cell–cell contact interactions between macrophages and dendritic T cells. An increase in the number of T cells and ED1-positive macrophages in the affected colonic tissue over time was documented. These results suggest colonic bacteria, host macrophages, and T cells play specific roles in the immunological reactions in DSS-induced colitis, possibly via an HSP60-mediated mechanism.     

Specific immunotherapy-induced Sjögren’s syndrome

Background: Allergen-specific immunotherapy (SIT) is a well-documented treatment for allergic rhinitis, asthma, and allergy to bee venoms. Side-effects of SIT in long-term have not been well documented yet. Herein, we report a case of Sjögrens syndrome following SIT. Case: The patient, a 25-year-old Caucasian woman, was started on subcutaneous grass-pollen immunotherapy. The patients autoantibodies before the SIT screening tests were negative. We determined that anti-extractable nuclear antigen (ENA) was positive (ENA = 98.4, normal range 0–25 U) on routine screening tests at 44 weeks of her treatment, and then SIT was discontinued. The patient complained of burning and itching in her eyes for 6 months. Schirmers and salivary flow tests were positive. Although antinuclear antigen and rheumatoid factor were negative, anti-SS-A/Ro was positive. Viral hepatitis markers were negative. Minor salivary-gland biopsy was performed, which showed grade 4 sialoadenitis. The HLA type of the patient was B55 (B22), Bw6, Cw1 for class I and DR11, DR52, DQ7 (DQ3) for class II. After the immunotherapy had been stopped, there were no changes in the symptoms and laboratory findings of the patient during the 1st year of follow-up. Conclusion: This is the first case to be reported of Sjögrens syndrome following SIT. Patients undergoing SIT must be carefully followed up for the development of autoimmunity and an autoimmune disease.     

Influence of TNF gene polymorphism in patients with acute and fulminant hepatitis

Background Tumor necrosis factor (TNF) is involved in liver damage, especially in fulminant hepatitis (FH). Our previous data showed that the serum level of TNF- was markedly increased in FH. To investigate the mechanism of the overproduction of TNF in FH patients, polymorphism of the TNF gene was studied.Methods We analyzed 120 healthy subjects (controls), 63 patients with acute hepatitis (AH), and 32 patients with FH. Of the 32 FH patients, 21 died or received liver transplantation (FH-D), and 11 survived with intensive therapy (FH-S). The TNF- promoter region at –1031, –863, –857, –308, and –238, and TNF- Nco1 polymorphism sites were studied.Results (1) The four groups showed no differences in polymorphisms of positions –857, –308, and –238. The allelic frequencies of positions –1031C and –863A in the FH-D patients were significantly higher compared to findings in control subjects. (2) The allelic frequency of B2 in the TNF- gene was significantly higher in FH patients, and particularly in the FH-D patients, compared to control subjects. (3) When the patients were divided into four groups by etiology, hepatitis A virus (HAV), HBV, HCV, and non-A non-B non-C, the allelic frequencies of positions –863A and TNF- B2 in FH patients were increased in the non-A non-B non-C group compared to controls.Conclusions FH patients with a poor prognosis had higher frequencies of positions –1031C and –863A in the TNF- promoter region, and higher frequencies of the B2 allele of the TNF- gene. These data suggest that the genomic background may be associated with the prognosis of acute liver failure.     

Increased interleukin 6 production by bronchoalveolar lavage cells in patients with active sarcoidosis

Alveolitis of sarcoidosis is characterized by activated alveolar macrophages (AMs) and T cells. The mediators interleukin-1 (IL-1) and interleukin 6 (IL-6) released by AMs represent essential factors for the progression of the T cells in the cell cycle. The role of IL-1 in pulmonary sarcoidosis has previously been studied; however, the relevance of other mediators (i.e. IL-6) has not yet been evaluated. We measured the spontaneous and lipopolysaccharide (LPS)-induced release of IL-6 and tumor necrosis factor a (TNF) by bronchoalveolar lavage cells (BAL) and peripheral blood mononuclear cells (PBMNC) in 6 control subjects (group A) and in 15 patients with sarcoidosis, 10 with active (group B), 5 with inactive disease (group C). IL-6 as well as TNF were spontaneously released by BAL cells of the active group in significantly greater amounts compared to both other groups; IL-6: A, 165.5 pg/ml/24 hr/10⁶ cells (range, 0–604), B, 946 (0–2467), C, 16.6 (0–83); TNF: A, 162 pg/ml/24 hr/10⁶ cells (0–523), B, 803 (100–17352), C, 100 (0–379). In all groups autologous PBMNC proved to be quiescent, releasing only baseline levels of the cytokines tested. After stimulation with LPS all these cells released great quantities of IL-6 and TNF. In active disease a positive correlation between IL-6 and TNF release was observed (r = 0.77, p < 0.02). The present study documents that in active sarcoidosis the spontaneous release of IL-6 by BAL cells parallels the spontaneous release of TNF. IL-6 is capable of initiating the proliferation and activation of T cells in the lung. Offprint requests to: J. Müller-Quernheim