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P.18 Amino acid infusion acutely increases circulating tumor necrosis factor in humans
decreased %PMN apoptosis at 12 hr (0 hr: GH 0 2.1 _+0.5 vs GH 100 2.5 _+ 1.3%, NS; 4hr: 18.9 4- 8.3 vs 10.3 4- 5.5%, NS; 12hr: 55.8 _+6.5 vs 30.5 _+ 7.2%, P < 0.05). %PMN apoptosis in surgical patients (n = 9) are shown in Table.
10 mg/kg, continuous 5 mg/kg.h; prime 30 mg/kg, continuous 5 mg/kg.h). Interleukin-6 (IL-6) was measured by B9-bioassay, hormones by radioimmunoassay and glucose production by primed continuous infusion of [6,6-2H2]glucose. Newman Keuls test was used for statistical analysis. Endotoxin induced fever, tachycardia and increased blood pressure (P < 0.05). Plasma IL-6 levels increased from 0.1 4- 0.1 to 27.4 _+6.6 ng/ml (P < 0.01). Plasma insulin levels increased from 6 4- 1 to 10 4- 2mU/I, glucagon from 29 4- 7 to 172 4- 75 rig/I, epinephrine from 0.42 _+ 0.14 to 0.78 4- 0.04 nmol/I and cortisol from 60 +_ 18 to 712 4- 82 nmol/] (P < 0.01 ). Hepatic glucose production increased from 13.7 _+ 1.6 to 23.6 43.3~mol/kg.min (P < 0.05). I-NMMA administration further increased the endotoxin-induced rise in blood pressure (P < 0.05 vs endotoxin alone). Endotoxin administration during I-NMMA infusion had no influence on the IL-6, neuro-endocrine and metabolic response to endotoxin. There were no differences in the response to endotoxin between the three different INMMAdoses. In conclusion, inhibition of nitric oxide production by high dose I-NMMA stimulates the increase in blood pressure in response to endotoxin, but does not affect the initial neuro-endocrine and metabolic responses to endotoxin. These data indicate that these responses to endotoxin are not merely mediated by nitric oxide induction.
Pre-opGH 0 GH100 POD1GH0 GH100 0hr 4.0_+1.3 3.5_+1.1 0.5-+0.2 3.2-+2.8 4 hr 6.6 -+4.0 4.6 -+2.5 5.8 _+3.7 8.0 _+6.4 12hr 61.1 -+10.4 50.8_+11.9" 21.0-+6.51 25.1 _+7.8t * P < 0.05 vs GH 0-12 hr, 1 P< 0.01 vs Pre-Op GH 0-12 hr, :~P< 0.05 vs Pre-Op GH 100-12 hr, by Student's t-test. Morphological findings also revealed that GH pretreatment resulted in a significant decrease in the percentage of PMN apoptosis in normal volunteers and in the preoperative samples at 12 hr culture (volunteers: GH 0 36.3 _+6.8 vs GH 100 18.0 _+5.5%, patients: 37.6 _+1.7 vs 23.0 +_7.2%, P < 0.01), but not in the samples isolated on POD 1 (21.4 _+8.1 vs 16.8 _+ 7.9%, NS). The percentage of PMN apoptosis at 12 hr culture without GH preincubation on POD 1 showed an inverse correlation with operative time (r = -0.72, P < 0.05) and with serum IL-8 levels on POD 1 (r = -0.61, P = 0.08). Conclusion: The results suggest that GH inhibits PMN apoptosis before surgery, but not in the early postoperative period. Surgical stress may blunt the inhibitory effects of GH on PMN apoptosis.
P.18 Amino acid infusion acutely increases circulating tumor necrosis factor in humans
P.16 lAPP induced anorexia in the rat can be partially reversed by an lAPP antagonist
G. Biolo, N. Toigo, C. Fiotti, C. Giansante, B. Ciocchi, G. Morena, R.
L. G r a n q v i s t 1'2, R. Reidelberger~, U. Amelo 2, T. E. Adrian 1, P.
Situlin and G. Guamieri Istituto di Clinica Medica, University of Trieste, Italy,
Westermark 3, D. Smith 1 and J. Permert 2 Dept of Biomed. Sci., Creighton Univ., Omaha, NE, USA and Depts of 2Surg., Karolinska Institute at Huddinge Univ. Hosp. and 3pathol., Univ. of Linkdping, Sweden.
Provision of total parenteral nutrition was associated with enhanced systemic and splanchnic production of cytokines. 1 This effect could be accounted for by alterations of the gastrointestinal integrity because of prolonged bowel rest or by direct effects of the intravenous nutrients on cytokine production. We have evaluated the acute effects of an intravenous amino acid administration on plasma concentrations of tumor necrosis factor-(z (TNF-o0 and of the p60 and p80 TNF-c~ soluble receptors (TNFsRp60 and TNFsRp80) in 8 healthy normal volunteers (male/female: 6/2; age: 38 4- 5 yrs; BM126 _+2 kg/m2). Subjects were studied in the basal postabsorptive state and at the end of a 3 hour primed (0.1 g.kg-1)-continuous (0.1 g.kg-l.h -~) infusion of a balanced amino acid mixture. Amino acid infusion increased (P = 0.01) plasma TNF-c~ concentrations from 5 4- 1 to 30 _+ 9 pg/ml. In contrast plasma TNF-c~ soluble receptors were not affected by amino acid infusion (TNFsRp60: from 598 4- 15 to 581 4- 15 pg/ml; TNFsRp80s from 300 4- 33 to 310 _+29 pg/ml). In conclusion, intravenous amino acid infusion increases TNF-c~ concentrations without affecting the TNF-c~ soluble receptors. This study provides evidence for a direct effect of parenteral amino acid administration on cytokine production.
We have previously reported elevated islet amyloid polypeptide in cachectic pancreatic cancer patients. Also, we have demonstrated that elevated plasma lAPP in the rat results in a marked reduction of food intake. The aim of the present study was to investigate the effect of 3 different lAPP antagonists (AC-187, lAPP 8-37, CGRP 8-37) on lAPPs food inhibitory effect. A specific antagonist should not only be able to block the effect of lAPP but also increase food intake when given alone. Indwelling jug. v. catheters were implanted in 16 male Wistar rats (275 g), individually housed and continuously monitodzed regarding food intake behaviour in an ad libitum fed model. The effect of each antagonist was tested in a cross-over experiment with four treatment groups (vehicle, lAPP, antagonist and lAPP + antagonist). Each rat received each treatment, infused under 3hours starting 15 minutes prior to the dark period = onset of 4-h test period, lAPP was delivered at 5 pmol/kg/min and the antagonists in a 100-fold higher dose. The following parameters were investigated: Cumulative food intake, Meal size, Meal number, Latency, Inter meal interval, Satiety ratio. lAPP caused a - 5 0 % inhibition of cumulative 4-h food intake (P < 0.001), and the effect was consistently reproducible. AC187 stimulated food intake during this period and had a maximal effect at 2 h, 3.5 _+0.6 g vs control 2.1 _+ 0.3 (P < 0.001), it partially blocked the effects of lAPP. lAPP(8.37) and CGRP(8-37) could neither inhibit the effects of lAPP except on latency nor stimulate FI. In this model AC187 was more effective than lAPP(8-37) and CGRP(8-37) and therefore appears to be more specific and is probably a true antagonist since food intake could be stimulated. Even though it is necessary to investigate the effect of higher doses, to try to completely block the effect of lAPP, the results are favouring that lAPP is a physiologic satiety factor.
Reference: 1. Ann Surg 1989; 210: 449-457.
P.19 Relationship between basal plasma leptin and cortisol levels in persons with various body fat content B. Krzyzanowska-~winiarska, M. ~miarowska, K. Pilarska. W. Wieliczko and S. Czekalski Dept of Endocrinology, Pomeranian Medical Academy, Szczecin, Poland.
M. R o b a c z y k ,
Leptin is a protein reflecting the size of adipose stores. Studies in rodents indicate that body fat content as well as leptin secretion is regulated by glucocorticoids, however data on this regulation in humans have been scanty so far. To elucidate the role of glucocorticoids in the regulation of leptin secretion we studied three groups of patients: A-BFD obesity, anorexia nervosa (AN) and Cushing's disease (CD) with different levels of endogenous cortisol and various body fat content. The study was performed in 11 women with ABF-D obesity (BMI 35.3 4- 1.7), 7 subjects with CD (BMI 27.2 4- 1.5) and 7 women with AN (BM114.8 4- 0.4). Basal plasma levels of leptin (LEP) and cortisol (C) were measured by RIA kits and body composition by DEXA. Adiposity-corrected plasma LEP levels were calculated as a ratio of plasma LEP to indices of body fatness: total body fat (BF), % body fat (%BF). Plasma C levels in subjects with AN and CD did not differ, but were significantly higher (P = 0.01 ) than in subjects with ABF-D obesity (727.2 _+ 62.9 and 893.1 4- 114.7 vs 574.9 4- 52.3 nmol/L, respectively). Subjects with AN had extremely low plasma LEP levels (2.95 _+0.4 ng/mL), while persons with ABF-D obesity and CD had comparable absolute plasma LEP levels (22.4 +_ 3.4 vs 23.0 4- 7.0 ng/ml, NS), though the obese had significantly
P,17 Inhibition of nitric oxide production by I-NMMA does not affect the initial neuro-endocrine and metabolic responses to endotoxin in dogs H. S. M o e n i r a l a m 1, F. Sprangers 1, E. Endert~, M. T. Ackermans ~, J. J. B. van Lanscho~, H. P. Sauerwein 1 and J. A. Romijn 1 Depts of ~Internal Medicine and 2Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. Ne-monomethyI-L-arginine (I-NMMA) inhibits endotoxin-induced nitric oxide production by inhibition of nitric oxide synthase. To investigate the influence of nitric oxide on neuro-endocrine and metabolic responses to endotoxin (E. coil, 1.0 #g/kg, i.v.), we studied six awake dogs on four different occasions: endotoxin alone, versus endotoxin during three different doses of I-NMMA (prime 10mg/kg, continuous 1 mg/kg.h; prime 27
10 mg/kg, continuous 5 mg/kg.h; prime 30 mg/kg, continuous 5 mg/kg.h). Interleukin-6 (IL-6) was measured by B9-bioassay, hormones by radioimmunoassay and glucose production by primed continuous infusion of [6,6-2H2]glucose. Newman Keuls test was used for statistical analysis. Endotoxin induced fever, tachycardia and increased blood pressure (P < 0.05). Plasma IL-6 levels increased from 0.1 4- 0.1 to 27.4 _+6.6 ng/ml (P < 0.01). Plasma insulin levels increased from 6 4- 1 to 10 4- 2mU/I, glucagon from 29 4- 7 to 172 4- 75 rig/I, epinephrine from 0.42 _+ 0.14 to 0.78 4- 0.04 nmol/I and cortisol from 60 +_ 18 to 712 4- 82 nmol/] (P < 0.01 ). Hepatic glucose production increased from 13.7 _+ 1.6 to 23.6 43.3~mol/kg.min (P < 0.05). I-NMMA administration further increased the endotoxin-induced rise in blood pressure (P < 0.05 vs endotoxin alone). Endotoxin administration during I-NMMA infusion had no influence on the IL-6, neuro-endocrine and metabolic response to endotoxin. There were no differences in the response to endotoxin between the three different INMMAdoses. In conclusion, inhibition of nitric oxide production by high dose I-NMMA stimulates the increase in blood pressure in response to endotoxin, but does not affect the initial neuro-endocrine and metabolic responses to endotoxin. These data indicate that these responses to endotoxin are not merely mediated by nitric oxide induction.
Pre-opGH 0 GH100 POD1GH0 GH100 0hr 4.0_+1.3 3.5_+1.1 0.5-+0.2 3.2-+2.8 4 hr 6.6 -+4.0 4.6 -+2.5 5.8 _+3.7 8.0 _+6.4 12hr 61.1 -+10.4 50.8_+11.9" 21.0-+6.51 25.1 _+7.8t * P < 0.05 vs GH 0-12 hr, 1 P< 0.01 vs Pre-Op GH 0-12 hr, :~P< 0.05 vs Pre-Op GH 100-12 hr, by Student's t-test. Morphological findings also revealed that GH pretreatment resulted in a significant decrease in the percentage of PMN apoptosis in normal volunteers and in the preoperative samples at 12 hr culture (volunteers: GH 0 36.3 _+6.8 vs GH 100 18.0 _+5.5%, patients: 37.6 _+1.7 vs 23.0 +_7.2%, P < 0.01), but not in the samples isolated on POD 1 (21.4 _+8.1 vs 16.8 _+ 7.9%, NS). The percentage of PMN apoptosis at 12 hr culture without GH preincubation on POD 1 showed an inverse correlation with operative time (r = -0.72, P < 0.05) and with serum IL-8 levels on POD 1 (r = -0.61, P = 0.08). Conclusion: The results suggest that GH inhibits PMN apoptosis before surgery, but not in the early postoperative period. Surgical stress may blunt the inhibitory effects of GH on PMN apoptosis.
P.18 Amino acid infusion acutely increases circulating tumor necrosis factor in humans
P.16 lAPP induced anorexia in the rat can be partially reversed by an lAPP antagonist
G. Biolo, N. Toigo, C. Fiotti, C. Giansante, B. Ciocchi, G. Morena, R.
L. G r a n q v i s t 1'2, R. Reidelberger~, U. Amelo 2, T. E. Adrian 1, P.
Situlin and G. Guamieri Istituto di Clinica Medica, University of Trieste, Italy,
Westermark 3, D. Smith 1 and J. Permert 2 Dept of Biomed. Sci., Creighton Univ., Omaha, NE, USA and Depts of 2Surg., Karolinska Institute at Huddinge Univ. Hosp. and 3pathol., Univ. of Linkdping, Sweden.
Provision of total parenteral nutrition was associated with enhanced systemic and splanchnic production of cytokines. 1 This effect could be accounted for by alterations of the gastrointestinal integrity because of prolonged bowel rest or by direct effects of the intravenous nutrients on cytokine production. We have evaluated the acute effects of an intravenous amino acid administration on plasma concentrations of tumor necrosis factor-(z (TNF-o0 and of the p60 and p80 TNF-c~ soluble receptors (TNFsRp60 and TNFsRp80) in 8 healthy normal volunteers (male/female: 6/2; age: 38 4- 5 yrs; BM126 _+2 kg/m2). Subjects were studied in the basal postabsorptive state and at the end of a 3 hour primed (0.1 g.kg-1)-continuous (0.1 g.kg-l.h -~) infusion of a balanced amino acid mixture. Amino acid infusion increased (P = 0.01) plasma TNF-c~ concentrations from 5 4- 1 to 30 _+ 9 pg/ml. In contrast plasma TNF-c~ soluble receptors were not affected by amino acid infusion (TNFsRp60: from 598 4- 15 to 581 4- 15 pg/ml; TNFsRp80s from 300 4- 33 to 310 _+29 pg/ml). In conclusion, intravenous amino acid infusion increases TNF-c~ concentrations without affecting the TNF-c~ soluble receptors. This study provides evidence for a direct effect of parenteral amino acid administration on cytokine production.
We have previously reported elevated islet amyloid polypeptide in cachectic pancreatic cancer patients. Also, we have demonstrated that elevated plasma lAPP in the rat results in a marked reduction of food intake. The aim of the present study was to investigate the effect of 3 different lAPP antagonists (AC-187, lAPP 8-37, CGRP 8-37) on lAPPs food inhibitory effect. A specific antagonist should not only be able to block the effect of lAPP but also increase food intake when given alone. Indwelling jug. v. catheters were implanted in 16 male Wistar rats (275 g), individually housed and continuously monitodzed regarding food intake behaviour in an ad libitum fed model. The effect of each antagonist was tested in a cross-over experiment with four treatment groups (vehicle, lAPP, antagonist and lAPP + antagonist). Each rat received each treatment, infused under 3hours starting 15 minutes prior to the dark period = onset of 4-h test period, lAPP was delivered at 5 pmol/kg/min and the antagonists in a 100-fold higher dose. The following parameters were investigated: Cumulative food intake, Meal size, Meal number, Latency, Inter meal interval, Satiety ratio. lAPP caused a - 5 0 % inhibition of cumulative 4-h food intake (P < 0.001), and the effect was consistently reproducible. AC187 stimulated food intake during this period and had a maximal effect at 2 h, 3.5 _+0.6 g vs control 2.1 _+ 0.3 (P < 0.001), it partially blocked the effects of lAPP. lAPP(8.37) and CGRP(8-37) could neither inhibit the effects of lAPP except on latency nor stimulate FI. In this model AC187 was more effective than lAPP(8-37) and CGRP(8-37) and therefore appears to be more specific and is probably a true antagonist since food intake could be stimulated. Even though it is necessary to investigate the effect of higher doses, to try to completely block the effect of lAPP, the results are favouring that lAPP is a physiologic satiety factor.
Reference: 1. Ann Surg 1989; 210: 449-457.
P.19 Relationship between basal plasma leptin and cortisol levels in persons with various body fat content B. Krzyzanowska-~winiarska, M. ~miarowska, K. Pilarska. W. Wieliczko and S. Czekalski Dept of Endocrinology, Pomeranian Medical Academy, Szczecin, Poland.
M. R o b a c z y k ,
Leptin is a protein reflecting the size of adipose stores. Studies in rodents indicate that body fat content as well as leptin secretion is regulated by glucocorticoids, however data on this regulation in humans have been scanty so far. To elucidate the role of glucocorticoids in the regulation of leptin secretion we studied three groups of patients: A-BFD obesity, anorexia nervosa (AN) and Cushing's disease (CD) with different levels of endogenous cortisol and various body fat content. The study was performed in 11 women with ABF-D obesity (BMI 35.3 4- 1.7), 7 subjects with CD (BMI 27.2 4- 1.5) and 7 women with AN (BM114.8 4- 0.4). Basal plasma levels of leptin (LEP) and cortisol (C) were measured by RIA kits and body composition by DEXA. Adiposity-corrected plasma LEP levels were calculated as a ratio of plasma LEP to indices of body fatness: total body fat (BF), % body fat (%BF). Plasma C levels in subjects with AN and CD did not differ, but were significantly higher (P = 0.01 ) than in subjects with ABF-D obesity (727.2 _+ 62.9 and 893.1 4- 114.7 vs 574.9 4- 52.3 nmol/L, respectively). Subjects with AN had extremely low plasma LEP levels (2.95 _+0.4 ng/mL), while persons with ABF-D obesity and CD had comparable absolute plasma LEP levels (22.4 +_ 3.4 vs 23.0 4- 7.0 ng/ml, NS), though the obese had significantly
P,17 Inhibition of nitric oxide production by I-NMMA does not affect the initial neuro-endocrine and metabolic responses to endotoxin in dogs H. S. M o e n i r a l a m 1, F. Sprangers 1, E. Endert~, M. T. Ackermans ~, J. J. B. van Lanscho~, H. P. Sauerwein 1 and J. A. Romijn 1 Depts of ~Internal Medicine and 2Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. Ne-monomethyI-L-arginine (I-NMMA) inhibits endotoxin-induced nitric oxide production by inhibition of nitric oxide synthase. To investigate the influence of nitric oxide on neuro-endocrine and metabolic responses to endotoxin (E. coil, 1.0 #g/kg, i.v.), we studied six awake dogs on four different occasions: endotoxin alone, versus endotoxin during three different doses of I-NMMA (prime 10mg/kg, continuous 1 mg/kg.h; prime 27