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Introduction Chronic allograft injury is the most frequent cause of long-term kidney allograft loss. Cyclosporine (CsA) nephrotoxicity is an important factor contributing to graft loss. Increased fibrosis has been reported in renal transplants under CsA as compared with tacrolimus (Tac). Platelet-derived growth factor (PDGF) is a well-characterized fibrogenic growth factor in renal disease. Objective Here we investigated the effect of Tac on kidney grafts and PDGF expression both early after transplantation as well as during long-term follow-up of rat renal allografts, and compared it to CsA. Tac and CsA were also studied on PDGF secretion in macrophages in vitro. Materials and methods Kidney allografts were immunosuppressed with Tac or CsA. Grafts were analyzed by histology and immunohistochemistry. ELISA was used to measure PDGF-levels in Tac and CsA-treated macrophage cultures. Results Tac ameliorated markedly both acute and chronic changes, whereas moderate to intense histological changes were seen in CsA-treated allografts. Tac also significantly inhibited PDGF expression compared to CsA. At clinically adjusted concentration CsA induced PDGF in macrophages whereas Tac did not. Discussion Tac may be less fibrogenic than CsA by decreasing PDGF expression in kidney grafts as well as in macrophages. Data presented here suggests that decreased PDGF induction by Tac may be beneficial for later outcome of the kidney graft.

13-cis-Retinoic acid treatment causes insulin resistance and disturbances in lipid and glucose metabolism. We studied how 13-cis-retinoic acid affects inflammatory factors and adiponectin. A total of 23 healthy patients (age, 24.9 +/- 0.9 years; body mass index, 22.6 +/- 0.7 kg/m(2)) who received 13-cis-retinoic acid treatment of acne participated in the study. The patients were studied before the treatment, after 3 months of therapy, and 1 month after the treatment. Inflammatory parameters were measured, and a 4-hour oral glucose tolerance test was performed at each visit. Treatment with 13-cis-retinoic acid resulted in a significantly elevated serum adiponectin concentration (from 24.9 +/- 2.5 to 29.4 +/- 3.6 mg/L, P < .05), hemoglobin A(1c) (from 5.27% +/- 0.05% to 5.42% +/- 0.06%, P < .01), C-peptide area under the curve (from 314.2 +/- 16.6 to 350.0 +/- 21.0 (nmol . min)/L, P < .05), and triglycerides (from 0.97 +/- 0.06 to 1.29 +/- 0.10 mmol/L, P < .05), whereas high-density lipoprotein cholesterol decreased (from 1.50 +/- 0.07 to 1.38 +/- 0.08 mmol/L, P < .05). The increase in adiponectin during 13-cis-retinoic acid therapy correlated with baseline triglycerides (r = 0.51, P < .02). Many inflammatory markers, which were nonsignificantly elevated during therapy, decreased significantly after cessation of treatment. These were C-reactive protein (median from 1.78 to 1.23 mg/L, P < .05), soluble intercellular adhesion molecule 1 (from 210 +/- 10 to 204 +/- 10 microg/L, P < .02), ceruloplasmin (256 +/- 17 to 231 +/- 17 microg/L, P < .02), and erythrocyte sedimentation rate (from 6.4 +/- 1.3 to 4.7 +/- 0.9 mm/h, P < .02). Interleukin 6 concentration was unaffected by the therapy, but decreased significantly after the treatment (from 2.18 +/- 0.46 to 1.65 +/- 0.43 ng/L, P < .05). In conclusion, although treatment with 13-cis-retinoic acid results in disturbances in glucose and lipid metabolism, paradoxically serum adiponectin concentration increases. 13-cis-Retinoic acid has profound effects on the regulation of inflammatory markers.

AIMS To investigate how reduction of hyperglycaemia affects acute phase serum proteins in poorly controlled type 1 diabetic patients. METHODS 24 patients (age 31.7 +/- 2.0 years, HbA1c 9.3 +/- 0.2%, BMI 24.2 +/- 0.7 kg/m2, diabetes duration 15.3 +/- 1.7 years) participated in the study. The treatment was optimised for 16 weeks. Blood samples were taken at baseline and at the end of the study. 16 healthy age- and BMI-matched subjects were chosen for a control group. RESULTS At baseline, the patients had higher C-reactive protein (CRP) (1.09, median [range 0.24-18.82] mg/l vs. 0.66 [0.18-2.46] mg/l, p < 0.02), mean adiponectin (16.06 +/- 1.31 vs. 8.85 +/- 0.93 mg/l, p < 0.001), ceruloplasmin (306 +/- 16.1 vs. 205.4 +/- 5.5 mg/l, p < 0.001), fibrinogen (3.41 +/- 0.26 vs. 2.38 +/- 0.07 g/l, p < 0.001), soluble intercellular adhesion molecule-1 (sICAM-1) (255.4 +/- 10.3 vs. 194 +/- 10.6 microg/l, p < 0.001), soluble vascular adhesion molecule-1 (sVCAM-1) (533.4 +/- 21.8 vs. 422.9 +/- 20.7 microg/l, p < 0.01) and interleukin-6 (2.89 +/- 0.49 vs. 1.35 +/- 0.30 ng/l, p < 0.01) concentrations than the controls. During intensified treatment, HbA1c decreased (to 8.5 +/- 0.2%, p < 0.001). This resulted in reduced sE-selectin (from 44.6 +/- 2.6 to 38.8 +/- 2.6 microg/l, p < 0.01), alpha-1-acid-glycoprotein (A1GP) (from 622.9 +/- 47.9 to 525.7 +/- 27.9 mg/l, p < 0.01), sICAM-1 (from 255.4 +/- 10.3 to 240.8 +/- 9.1 microg/l, p < 0.05) and IL-6 (from 2.9 +/- 0.5 to 2.1 +/- 0.4 ng/l, p < 0.01). Serum amyloid A (SAA) and CRP did not change 12.00 (0.7-222.0) vs. 12.00 (1.6-277.0) mg/l for SAA and 1.09 (0.24-18.82) vs. 1.09 (0.18-23.08) mg/l for CRP, baseline vs. treatment, respectively. CONCLUSIONS Poorly controlled type 1 diabetic patients have increased values of adiponectin, CRP, ceruloplasmin, fibrinogen, sICAM-1, sVCAM-1 and IL-6. Reduction of hyperglycaemia results in decreased sE-selectin, A1GP, sICAM-1 and IL6, while other inflammatory factors including CRP, SAA and adiponectin are not affected.

OBJECTIVE—To elucidate whether serum adiponectin is associated with renal function, low-grade inflammatory markers, metabolic control, and insulin resistance in type 1 diabetic patients with and without nephropathy. RESEARCH DESIGN AND METHODS—A total of 189 type 1 diabetic patients from the Finnish Diabetic Nephropathy Study were divided into three groups based on their urinary albumin excretion rate (AER): patients with normal AER (n = 66) had no antihypertensive medication, while patients with microalbuminuria (n = 63) or macroalbuminuria (n = 60) were all treated with an ACE inhibitor. Renal function was estimated with the Cockcroft-Gault formula. Adiponectin was measured by an in-house time-resolved immunofluorometric assay. RESULTS—Adiponectin concentrations were higher in women than in men, but since there was no significant difference in sex distribution between the groups, data were pooled. Adiponectin concentrations were higher in patients with macroalbuminuria (19.8 ± 12.0 mg/l) than in patients with microalbuminuria (13.1 ± 4.8 mg/l) or normoalbuminuria (11.8 ± 4.2 mg/l). In a univariate analysis, adiponectin was positively associated with creatinine (r = 0.41; P < 0.0001), AER (r = 0.33; P < 0.0001), interleukin-6 (r = 0.22; P = 0.002), systolic blood pressure (r = 0.22; P = 0.004), HbA1c (r = 0.17; P = 0.02), total cholesterol (r = 0.16; P = 0.03), and HDL cholesterol (r = 0.16; P = 0.03) and negatively with estimated glomerular filtration rate (GFR; r = −0.52; P < 0.0001) and waist-to-hip ratio (WHR; r = −0.16; P = 0.03). In a multiple linear regression analysis including the above variables, estimated GFR, AER, and WHR were independently associated with adiponectin levels (r2 = 0.32). CONCLUSIONS—Serum adiponectin concentrations are increased in type 1 diabetic patients with nephropathy, and levels are further associated with renal insufficiency.

Background: Glucose absorbed from conventional peritoneal dialysis (PD) solutions contributes to unfavorable metabolic effects. Its replacement with a glucose-free osmotic agent such as icodextrin (ID) or amino acids (AA) may have some benefit on glucose and lipid metabolism. Methods: Serum lipids, insulin sensitivity and substrate oxidation (calorimetry) were measured before and after 8 weeks use of ID or AA in 22 patients. Calorimetry and blood tests (HbA1c, lipids) were also performed after 8 weeks of simultaneous use of ID and AA in 8 patients. Results: Cholesterol declined during the use of AA (4.8 ± 0.3–4.5 ± 0.3 mmol/l, p = 0.045). Triglycerides decreased during the use of both ID (2.2 ± 0.2–1.9 ± 0.1 mmol/l, p = 0.019) and AA (1.9 ± 0.2–1.6 ± 0.1 mmol/l, p = 0.024). Free fatty acids declined during the use of AA. There were no significant changes in insulin sensitivity. Glucose oxidation decreased and lipid oxidation increased during the use of ID, the changes in substrate oxidation were accentuated during the simultaneous use of ID and AA. Conclusion: Replacement of glucose with ID or AA had a benefit on glucose and lipid metabolism.

Diabetic nephropathy is associated with insulin resistance, and low-grade inflammation and activation of the complement system may contribute to this cascade. Mannan-binding lectin (MBL) activates the complement system, and elevated MBL concentrations have been observed in normoalbuminuric type 1 diabetic patients. The aim of this study was to assess whether MBL is associated with diabetic nephropathy in type 1 diabetes, and whether there is an association between MBL and low-grade inflammatory markers or insulin resistance.A total of 191 type 1 diabetic patients from the Finnish Diabetic Nephropathy Study were divided into three groups based upon their AER. Patients with normal AER (n=67) did not take antihypertensive medication, while patients with microalbuminuria (n=62) or macroalbuminuria (n=62) were all treated with an ACE inhibitor. As a measure of insulin sensitivity we used estimated glucose disposal rate. MBL was measured by an immunofluorometric assay, C-reactive protein by a radioimmunoassay and IL-6 by high-sensitivity enzyme immunoassay.Patients with normal AER (median [interquartile range]: 1,154 microg/l [180-2,202 microg/l]) had lower levels of MBL than patients with microalbuminuria (1,713 microg/l [724-2,760 microg/l]; p=0.029) or macroalbuminuria (1,648 microg/l [568-3,394 microg/l]; p=0.019). There was a significant correlation between MBL and estimated glucose disposal rate, but not between MBL and C-reactive protein or IL-6 levels in univariate analysis. However, in a multiple regression analysis, HbA1c was the single variable independently associated with MBL (beta+/-SEM: 0.26+/-0.08; p=0.003).MBL concentrations are increased in type 1 diabetic patients with diabetic nephropathy. MBL was not associated with low-grade inflammatory markers.

BACKGROUND Albumin and alpha1-microglobulin (alpha1M) are absorbed by two specific receptors in tubular epithelial cells. Any cell injury will disturb the reabsorption of these proteins, The increased urinary excretions of albumin or alpha1M could thus serve as a marker of subclinical graft lesions and as an early indicator of chronic allograft dysfunction. METHODS We measured 24-hour urinary excretions of albumin, alpha1M, and transforming growth factor (TGF)-beta1 at 6 months after transplantation in 79 renal-graft recipients and recorded the changes in 24-hour creatinine clearance an average 51 (range 14-72) posttransplant follow-up months. RESULTS At 6 months from transplantation, 46 of 79 (58%) patients were normoalbuminuric, 25 (32%) microalbuminuric, and 8 (10%) macroalbuminuric. In normoalbuminuric patients, urinary alpha1M/creatinine ratio was 10 times, and TGF-beta1/creatinine ratio approximately 5 times, higher than in the healthy subjects but lower than in albuminuric patients. In all patients, urinary alpha1M correlated with urinary TGF-beta1 (r=0.508, P

Background/Aims: Despite effective antibiotic therapy, peritonitis still remains a major problem in peritoneal dialysis (PD). The aim of the present study was to investigate changes of CRP, dialysate leukocytes and IL-6, hyaluronan (HA) and sICAM-1 in dialysate during and after peritonitis and their association to the outcome of peritonitis. Methods: Dialysate IL-6, HA and sICAM-1 were measured at the onset and on day 4, at the end of the treatment and 2 months after onset of peritonitis. Furthermore, CRP and dialysate leukocytes were measured on days 1–4. Results: All measured soluble factors were higher on the first and fourth day than at the end of the treatment. sICAM-1 and HA were lower at the end of the treatment in patients who later had a relapse/re-infection. IL-6 remained higher 2 months after clinically cured peritonitis. CRP and dialysate leukocytes were higher on day 4 in patients with poor outcome. Conclusions: Peritonitis causes increased excretion of soluble factors. Low concentrations of sICAM-1 and HA at the end of the treatment were negative prognostic indicators. Higher IL-6 levels after peritonitis could be a sign of ongoing inflammation in the peritoneal membrane. Delayed decrease in CRP and dialysate leukocytes may indicate poor outcome.

BACKGROUND IgA nephropathy (IgAN) and Henoch-Schonlein nephritis (HSN) share many clinical, histological and immunological features. It has been postulated that these two conditions have a common pathogenesis and that HSN might be a systemic form of IgAN. Activity of interleukin-1beta (IL-1beta) in urine has been found to be higher in IgAN and HSN patients than in healthy controls. Interaction between IL-1beta and interleukin-1 receptor antagonist (IL-1ra) plays a significant role in the regulation of inflammatory responses. We studied levels of urinary excretion of IL-1beta and IL-1ra in patients with IgAN and HSN. METHODS Amounts of IL-1beta and IL-1ra excreted in 24-h urine samples collected from 241 IgAN, 26 HSN patients and from 33 healthy controls were determined. Results were expressed as cytokine/creatinine (ng/mmol) ratios. RESULTS Urinary IL-1beta excretion by the IgAN and HSN patients was no greater than urinary IL-1beta excretion by healthy controls. Urinary IL-1ra excretion by the IgAN patients was lower than urinary IL-1ra excretion by healthy controls (P < 0.05) and by the HSN patients (P < 0.01). In both patients and controls women had significantly higher IL-1ra, IL-1beta excretion levels and IL-1ra/IL-1beta ratios. The differences in urinary excretions of IL-1ra by the healthy controls and by the IgAN and HSN patients were significant in both sexes. Excretion of IL-1beta or IL-1ra did not correlate with excretion of urinary protein, duration of the disease or any histopathological variable. However, histopathological changes in renal biopsy specimens from patients with IL-1ra/IL-1beta ratios above normal were significantly milder than in renal biopsy specimens from patients with low or normal IL-1ra/IL-1beta ratios. CONCLUSION Urinary IL-1ra levels in IgAN patients were lower than urinary IL-1ra levels in healthy controls or HSN patients, a finding which may indicate that the two diseases have a different pathogenesis. Whether the male predominance in IgAN and HSN and the worse outcomes in males that have been reported previously in IgAN and HSN are connected with the lower excretion of IL-1ra and consequently lower IL-1ra/IL-1beta ratios in male patients than in female patients needs more thorough investigation.

The bioincompatibility of dialysis solutions and recurrent episodes of peritonitis may alter peritoneal function. Cytokines and growth factors may play a role in inflammatory and fibrotic processes. We therefore investigated whether there is a correlation between peritoneal function and excretion of cytokines and other soluble factors in dialysate.A personal dialysis capacity test was performed in 40 stable peritoneal dialysis patients. Tumour necrosis factor-alpha, interleukin-6 (IL-6), hyaluronan, soluble intercellular cell adhesion molecule-1 and transforming growth factor-beta1 were analysed from overnight and 24-h dialysates during the test.We found little evidence for a direct correlation between cytokines and other soluble factors in dialysate and dialysis adequacy. There was, however, a strong correlation between the measured soluble factors and characteristics of the peritoneal membrane. Furthermore, IL-6 correlated with the number of previous episodes of peritonitis.Soluble factors in dialysate may indicate ongoing inflammatory processes in the peritoneal membrane, which may gradually lead to alterations in peritoneal function.

Background. Intercellular adhesion molecule-1 (ICAM-1) binds to leukocyte adhesion receptors LFA-1 and MAC-1, and mediates leukocyte adhesion to target structures. During acute rejection there is increased expression of ICAM-1 in vascular and tubulointestial cells, and consequently accumulation of inflammatory leukocytes. Soluble ICAM-1 (sICAM-1) is released from ICAM-1 expressing cells and excreted into the surrounding fluid. Increased serum sICAM-1 levels are found in patients with acute rejections of various allografts, and high urinary levels in steroid resistant acute kidney allograft rejection. Methods. Urinary excretion of sICAM-1 was measured by EIA in 136 kidney allograft recipients during the first 1‐ 6 post transplant weeks: 30 patients developed acute rejection, and 106 patients had stable graft function. The molecular weight, binding to hyaluronan, and the origin of urinary sICAM-1 were studied. Results. We show that urinary sICAM-1 circulates as a monomer with a molecular weight between 50 and 100 kD. It binds to immobilized, but not to circulating hyaluronan. About one week after transplantation the mean sICAM-1/creatinine ratio (306 ng/mmol) in transplanted patients was higher than in the healthy controls (167 ng/mmol, P

Type 2 diabetes is characterized by increased acute phase serum proteins. We wanted to study how these proteins are related to complement activation in type 2 diabetes and how improvement of glycemic control affects them or complement activation. A total of 29 type 2 diabetic patients (age, 55.2 +/- 1.8 years, glycosylated hemoglobin [HbA(1c)] 8.9% +/- 0.2%, body mass index [BMI] 30.9 +/- 0.8 kg/m(2), duration 5.9 +/- 1.3 years) participated in the study. They were previously treated either with diet alone or in combination with 1 oral antihyperglycemic medication. After a period of at least 4 weeks run-in on diet only, the patients were randomized to pioglitazone, glibenclamide, or placebo. Blood samples were taken before the treatments and at the end of the 6-month therapy. Basal C-reactive protein (CRP) level was related to acylation-stimulating protein (ASP) concentration (r =.55, P

To analyze whether complement C3a anaphylatoxin, other C3 fragments, interleukin-1beta (IL-1beta), or tumor necrosis factor-alpha (TNF-alpha) contributes to inflammation in chronic otitis media with effusion (OME).The amount of C3a was measured by enzyme-linked immunoassay. Further breakdown of C3 was analyzed by Western blotting. IL-1beta and TNF-alpha concentrations were measured by radioimmunoassay. Bacteria were analyzed by culture and polymerase chain reaction.Highly elevated levels of C3a and other C3 cleavage fragments were found in all middle ear effusion (MEE) samples. The mean values (+/- SEM, n = 26) for C3a, IL-1beta, and TNF-alpha were 5,973 +/- 1,124 ng/mL, 1,043 +/- 490 pg/mL, and 79 +/- 14.3 pg/mL, respectively. Comparison to an average C3 level of 555 (+/-108) microg/mL indicated that at least 40.5% +/- 6% of total C3 had become activated within the MEE. C3a concentrations were higher in the group in which the effusion had been present in the middle ear for a prolonged period (or =4 mo) (P = .04). Children with multiple tube insertions had higher C3 (P = .006) and TNF-alpha (P = .04) concentrations in their MEE samples than those receiving their first tubes. C3 and C3a concentrations in MEE correlated to each other (correlation coefficient [r] = 0.513, P = .0056), as did concentrations of IL-1beta and TNF-alpha (r = 0.7016, P.0001). No significant correlation was found between complement C3 or C3a levels and IL-1beta, TNF-alpha, or bacterial growth.Highly elevated levels of C3a in MEE indicate ongoing complement activation, which is stronger than in almost any other disease demonstrated previously. Elevated C3a levels contribute to chemotactic and inflammatory potential in the MEE and correlate with the chronicity of the disease.

Background: Erythropoietic protoporphyria (EPP) is an inherited disease caused by deficient activity of ferrochelatase in the heme biosynthetic pathway. Accumulation of protoporphyrins and light exposure results in acute phototoxic skin reactions. The histopathologic findings of the light-exposed skin are thickening of the superficial dermal vessel walls and amorphous deposits around the vessels, but the origin and detailed composition of the perivascular material have been unclear. Objective: The vascular morphology and composition of the perivascular material were studied in the skin samples of patients with EPP. Methods: Skin biopsy specimens of 8 patients with EPP representing 7 Finnish EPP families with different genotypes were studied by means of light and electron microscopy and immunohistochemical methods. Results: The characteristic finding was thickened, periodic acid-Schiff-positive vessel walls caused by concentric reduplication of basal lamina and excess of fine granular material at the basal membrane zone in the superficial dermis. The perivascular deposits in the vicinity of vessel walls had a homogeneous or fine granular appearance without filaments. Direct immunofluorescence showed constant IgG deposits together with IgA, IgM, and C3 in the vessel walls. In immunohistochemistry, collagen IV and laminin could be demonstrated at the vascular basal membrane together with serum amyloid P protein, κ and λ light chains, and a 90-kd glycoprotein. Conclusion: The vascular involvement indicates that the blood vessel walls in the papillary dermis are the primary tissues affected during an acute photoreaction. The repeated acute damage and repair processes in the basement membrane zone result in thickening of the vessel walls. Perivascular deposits are a secondary and irreversible phenomenon resulting from the leakage and accumulation of different serum components. These changes were not found in the nonexposed skin, indicating that an increased level of erythrocyte protoporphyrin per se is not responsible for the cutaneous manifestations, but the interaction of solar radiation is mandatory. Amorphous deposits distinguish EPP from variegate porphyria and porphyria cutanea tarda; a histopathologic examination may be a helpful tool in differentiating porphyric and nonporphyric photosensitivity. (J Am Acad Dermatol 2000;43:489–97.)

Aims/hypothesis. Inflammation could play a part in insulin resistance. Thiazolidinediones, new antidiabetic drugs, possess anti-inflammatory effects in vitro. We investigated if acute-phase serum proteins are increased in patients with Type II (non-insulindependent) diabetes mellitus who had been treated with insulin and whether troglitazone has anti-inflammatory effects in vivo. Methods. A total of 27 patients (age 63.0 ‐ 1.7 years, HbA 1c 8.8 ‐ 0.3 %, BMI 32.7 ‐ 0.8 kg/m 2 , duration 15.2 ‐ 1.4 years, insulin dose 73.3 ‐ 7.0 U/day) participated in the study. The patients received daily either 400 mg troglitazone or placebo for 16 weeks. Blood samples were taken at baseline, at the end of therapy and after a follow-up time of 23 ‐ 4 days. Results. The concentrations of serum amyloid A (6.2 ‐ 1.1 mg/l) and C-reactive protein (6.1 ‐ 1.1 mg/ l) were increased (p < 0.001) and complement protein C3 (1.69 ‐ 0.05 g/l) was also above the reference range for healthy subjects. Placebo treatment had no effect on glucose or inflammation, whereas troglitazone reduced fasting glucose (from 10.4 ‐ 0.6 mmol/l to 8.1 ‐ 0.5 mmol/l, p < 0.01), HbA 1c (from 8.7 ‐ 0.3 % to 7.5 ‐ 0.3 %, p < 0.01), insulin requirements (from 75 ‐ 10 U/day to 63 ‐ 10 U/day, p < 0.05), serum amyloid A (from 6.3 ‐ 1.5 mg/l to 4.0 ‐ 1.3 mg/l, p = 0.001), a-1-acid glycoprotein (from 906 ‐ 51 mg/l to 729 ‐ 52 mg/l, p = 0.001) and C3 (from 1.72 ‐ 0.07 g/l to 1.66 ‐ 0.06 g/l, p < 0.05) but not a-1-antitrypsin, ceruloplasmin, C-reactive protein or haptoglobin significantly. Concentrations of glucose and acute-phase reactants had returned to those before treatment at the follow-up visit. Conclusion/interpretation. In Type II diabetic patients serum amyloid A and complement protein C3 are raised. Troglitazone exerts a selective reversible action on some acute-phase proteins and C3 but not on others in conjunction with the improvement in glucose metabolism. [Diabetologia (1999) 42: 1433‐ 1438]

Adhesion molecules and urinary tumor necrosis factor-α in idiopathic membranous glomerulonephritis. Adhesion molecules are required in several physiological processes, but their altered function/expression is associated with the pathogenesis of inflammatory diseases. In the present study on idiopathic membranous glomerulonephritis (MGN) the expression of adhesion molecules (ICAM-1, VCAM-1, PECAM-1, E-selectin, LFA-1, Mac-1) was analyzed in different cellular compartments of the kidney using an indirect immunoperoxidase technique and monoclonal antibodies. Relationships between the expression of these molecules and the clinical and morphological activity of the disease and the urinary excretion of tumor necrosis factor α (TNF-α) were studied in 20 patients. The results were compared with the findings in ten normal kidneys and urinary TNF-α in 17 healthy subjects. The expression of adhesion molecules in glomeruli and tubules was unchanged apart from a diminished expression of VCAM-1 (P = 0.014) in glomerular parietal epithelial cells and PECAM-1 in glomerular endothelial cells (P < 0.01). Interstitial peritubular capillaries expressed significantly (P = 0.009) more E-selectin compared with the controls. The interstitial compartment had a highly increased number of cells expressing ICAM-1 in MGN (32.4 ± 4.6 cells/high power field) compared with the controls (9.4 ± 1.2; P < 0.001). Also, cells expressing VCAM-1 (10.2 ± 1.6 vs. 2.8 ± 1.9; P = 0.005), PECAM-1 (25.9 ± 5.3 vs. 7.4 ± 2.1; P = 0.006), and LFA-1 (20.4 ± 3.6 vs. 8.3 ± 1.5; P = 0.041) were increased in the interstitium. Proteinuria correlated particularly with the expression of E-selectin in peritubular capillaries (r = 0.63, P = 0.004). The number of LFA-1 expressing inflammatory cells in the interstitium correlated with peritubular capillary E-selectin (r = 0.8, P < 0.001) and interstitial ICAM-1 (r = 0.61, P = 0.009) expression, but histological alterations did not correlate with the expression of adhesion molecules. Tumor necrosis factor-α excretion was significantly increased in MGN (41 ± 8 pg/mg creatinine) compared with the controls (13 ± 2; P = 0.001), and in particular, it correlated with the interstitial expression of LFA-1 (r = 0.71, P = 0.002). This study suggests that active MGN leads not only to proteinuria but also to increased urinary TNF-α excretion. These may serve as triggers for the up-regulation of adhesion molecules in the peritubular capillaries and interstitial cells thus enhancing the development of the interstitial injury.

To measure the pre- and postoperative tear fluid tumour necrosis factor-alpha (TNF-alpha) concentration and release in patients undergoing excimer laser photorefractive keratectomy (PRK).Tear fluid samples from 18 PRK patients were collected with scaled microcapillary tubes preoperatively (day 0), on the second (day 2), and on the seventh (day 7) postoperative days. The TNF-alpha concentration was measured using a double antibody radioimmunoassay, and the TNF-alpha release was calculated by multiplying the concentration by the tear fluid flow in the collection capillary.The mean tear fluid flow in the capillary was 22.5 microliters/min (range 1.5-93.2) on day 0, 80.7 microliters/min (3.0-219, p = 0.0002) on day 2, and 14.6 microliters/min (1.8-41.7, NS) on day 7. The mean TNF-alpha concentration and release values were: day 0, 358 ng/I (110-680) and 9.5 pg/min (0.2-37.5, NS); day 2, 417 ng/l (5-750, NS) and 28.6 pg/min (0.6-81.5; p = 0.003); and day 7, 320 ng/I (5-735, NS) and 4.8 pg/min (0-25.4, NS), respectively.TNF-alpha appears to be a component of normal tear fluid. In spite of hypersecretion caused by the corneal wound, TNF-alpha concentrations remain constant during wound healing. TNF-alpha release increases significantly during the 2 postoperative days following PRK, suggesting a role in corneal wound healing.

To study the presence and release of tear fluid transforming growth factor-beta 1 (TGF-beta 1) and vasoendothelial growth factor (VEGF) of 19 patients undergoing excimer laser photorefractive keratectomy.Tear fluid samples were collected with scaled microcapillary tubes preoperatively (Day 0) and on the first or second (Day 1-2) and seventh (Day 7) postoperative days. Since postoperative tear fluid flow was significantly higher, we calculated the release of both growth factors by multiplying the concentration by the tear fluid flow in the capillary.The median tear fluid flow in the capillary was 8.1 microliters/ min (range 0.7-20.8) on Day 0, 46.3 microliters/min (3.1-135.1) on Day 1-2, and 10.2 microliters/min (1.8-65) on Day 7. The median TGF-beta 1 concentration and release values were: Day 0: 4.5 ng/ ml (0-23) and 24 pg/min (0-257), Day 1-2: 9 ng/ml (0-32) and 446 pg/min (0-4602; p = 0.0014), and Day 7: 6.5 ng/ml (0-44) and 53 pg/min (0-470), respectively. The median VEGF concentrations and releases were: Day 0: 19 ng/ml (14-23) and 128 pg/min (11-458), Day 1-2: 15 ng/ml (11-20) and 695 pg/ min (14-2567; p = 0.0007), and Day 7: 17.5 ng/ml (15-22) and 167 pg/min (27-1235), respectively.TGF-beta 1 and VEGF are components of normal tear fluid. The significantly increased release of both growth factors after exeimer laser photorefractive keratectomy may influence the corneal wound healing process.

Urinary excretion of protectin (CD59), complement SC5b-9 and cytokines in membranous glomerulonephritis. Protectin (CD59) is a low molecular weight glycophosphoinositol-anchored inhibitor of the membrane attack complex of complement (MAC) that is present, for example, on the membranes of endothelial cells and on epithelial cells of glomeruli and distal tubuli. To examine for the possibility that CD59 becomes detached from cell surfaces following cell injury, this study evaluated renal excretion of CD59 in patients with idiopathic membranous glomerulonephritis (MGN; N = 21), diabetic nephropathy (DNP; N = 15) and in healthy control subjects (N = 13). CD59 in human urine was quantitated by a competitive solid-phase radioimmunoassay having ≈13 kDa soluble urinary CD59 as a standard. Immunofluorescence microscopy demonstrated a decreased expression of CD59 in the glomeruli of MGN patients. Using a Triton X-114 phase separation method 91 to 97% of urinary CD59 was found to be in a soluble form without anchor-associated phospholipid. The mean (±sem) level of urinary CD59 was 5.6 ± 0.2 µg/ml in MGN patients, 3.7 ± 0.4 µg/ml in healthy controls (P < 0.001) and 2.6 ± 0.1 in DNP patients (P < 0.001). When related to urinary creatinine (UCr) the corresponding values were 11.9 ± 5.6, 4.8 ± 0.3 (P = 0.021) and 4.4 ± 0.2 (P < 0.002), respectively. The amount of CD59 in urine correlated with the urinary excretion of soluble terminal complement complexes, SC5b-9 (r = 0.594, P < 0.006) in MGN patients. The excretion of CD59 also correlated with the excretion of the inflammatory mediator IL-1β (r = 0.671, P = 0.001) but not with TNF-α (r = 0.314, P = 0.178). No correlation of CD59 excretion was observed with duration of the disease, level of proteinuria, serum albumin concentration or serum creatinine level. Based on these findings we speculate that the increased excretion of CD59 into urine in MGN patients is due to complement activation and inflammation induced shedding of CD59 from glomerular cells.

The serum transferrin receptor (TfR) level reflects iron status and the rate of erythropoiesis. This study was undertaken to assess the role of serum TfR in the iron status and erythropoiesis in very-low-birth-weight infants under conditions in which erythropoiesis is stimulated by large doses of recombinant human erythropoietin (rHuEPO) and oral iron. The first 34 infants were followed from the 3rd to 11th wk of life or until discharged. They received iron at a rate of 3 mg/kg/d. The subsequent 21 infants were given rHuEPO (300 U/kg three times a week s.c.) and iron at a rate of 6 mg/kg/d from the 3rd or 4th wk of life for a mean of 3.4 wk. With this treatment, the need for transfusion was reduced from 1.4 ± 0.4 to 0.1 ± 0.1 transfusions per infant (p = 0.02). The serum TfR concentrations in the rHuEPO-treated infants increased gradually to values several-fold higher than those in the untreated infants. This increase was not related to intrauterine or postnatal growth, protein intake, or serum albumin concentration. Neither was an association observed between Hb and TfR concentration. In the treated infants, the serum ferritin concentration was lower at the 4th, 5th, and 7th wk of life than in the untreated infants. The very-low-birth-weight infants who were given large doses of rHuEPO and iron had a marked rise in serum TfR concentration and a small decline in serum ferritin concentration. These events have been related to iron deficiency.

Liu J, Kahri AI, Teppo A-M, Voutilainen R. Regulation of parathyroid hormone gene expression and peptide secretion in human parathyroid cells. Eur J Endocrinol 1994;130:394–401. ISSN 0804–4643 In cell cultures prepared from human parathyroid adenomas, parathyroid hormone (PTH) mRNA expression decreased slowly. During short-term incubations (less than 24), a low calcium concentration (0.5 mmol/l) and protein kinase C activator TPA (12-O-tetradecanoyl phorbol 13-acetate) (160 nmol/l) increased PTH secretion (60%; p Raimo Voutilainen, Department of Pathology, PO Box 21 (Haartmaninkatu 3), SF-00014 University of Helsinki, Finland

We examined the acute phase reaction in myocardial infarction after thrombolytic treatment by streptokinase or tissue plasminogen activator. The magnitude of the acute phase reaction as determined by measurements of serum C-reactive protein and amyloid-A protein did not correlate with infarct size (determined by serial measurements of creatine kinase-MB) in this patient population. On the other hand, the development of acute cardiac failure was more closely associated with the magnitude of the acute phase reaction than with infarct size. The peak serum values of C-reactive protein in patients with and without acute cardiac failure were 128 mg/l (95% confidence intervals 85-170) and 60 mg/l (30-89); P less than 0.01 and concentration time integrals 578 mg/l x days (368-787) and 205 mg/l x days (62-350); P less than 0.01. The corresponding creatine kinase-MB values were 310 U/l (191-429) and 207 U/l (125-289) not significant; and 319 U/l x days (201-437) and 204 U/l x days (124-286) not significant; respectively. Patients requiring medication for cardiac failure on discharge from hospital had higher C-reactive protein and serum amyloid A protein values than those who did not, although the difference did not quite reach statistical significance. The infarct sizes were similar whether the patients needed medication for cardiac failure at discharge or not. Subjectively felt morbidity due to myocardial infarction was linearly associated with serum C-reactive protein peak values (P less than 0.05) and concentration time integrals (P less than 0.05), but not with infarct size. We conclude that thrombolytic treatment of myocardial infarction may reduce hospital inpatient morbidity independently of the limitation of infarct size. This diminished morbidity seems to be associated with modest or low acute phase reaction.

Urinary excretion of kappa light chains in patients with diabetes mellitus. The urinary excretion of kappa light chains,β 2 -microglobulin and albumin was examined in patients with newly diagnosed and long-standing insulin-dependent (IDDM) and non-insulin-dependent (NIDDM) diabetes mellitus, and compared to age-matched control subjects. Patients with IDDM diagnosed within two months, presented with normal albumin excretion, whereas the concentrations of β 2 -microglobulin and kappa light chain in urine were higher than in control subjects. The initiation of insulin therapy reduced, but did not completely normalize, the elevated rate of kappa light chain excretion. Patients with IDDM of long duration showed increased urine excretion of kappa light chains and albumin. In keeping with the findings in IDDM, patients with newly diagnosed NIDDM (within one year) showed increased urinary excretion of kappa light chains compared with control subjects. There was, however, no further increase in light chain excretion with longer duration of NIDDM. To study the effect of short-term hyperglycemia on urinary protein excretion, 12 normal subjects participated in a three-step hyperglycemic clamp study, during which their plasma glucose concentration was raised by +50, +125 and +300 mg/dl. The urine excretion of albumin and β; 2 -microglobulin rose progressively with each hyperglycemic clamp step, whereas that of kappa light chain excretion was unaffected by hyperglycemia. We conclude that increased urinary excretion of kappa light chain is a consistent finding in all types of diabetes mellitus, and can be observed even when the albumin excretion is normal. Since the serum concentration of kappa light chain is normal in diabetes, the increased urinary excretion of kappa light chains must be of renal origin. Although the underlying mechanism(s) responsible for the kappa light chainuria are unknown, it cannot be ascribed to the effect of acute hyperglycemia on glomerular and tubular function.

Background: Type 2 diabetes is characterized by increased acute phase serum proteins. They are also risk factors for cardiovascular disease. We wanted to study how improvement of glycemic control with pioglitazone or glibenclamide affects their serum concentrations. Materials and Methods: A total of 59 patients with Type 2 diabetes (age 57.3±1.2 yr, glycosylated hemoglobin (HbA1c) 8.3±0.7%, body mass index (BMI) 31.4±0.8 kg/m2) participated in the study. They were previously treated either with diet alone or in combination with one oral antihyperglycemic medicine. After a 1-week lead-in period on diet only, the patients were randomized to pioglitazone or glibenclamide. Blood samples for α-1-acid glycoprotein (A1GP), C-reactive protein (CRP) and serum amyloid A (SAA) were taken before the treatments and during the therapy after 20 and 52 weeks. Results: Baseline A1GP correlated with CRP (r=0.70, p

Background Inflammation is an established contributor in atherosclerosis and several other common diseases including diabetes. Therefore the study was to investigate how inflammatory factors affect lipid metabolism during recovery from hyperinsulinaemia in healthy individuals. Materials and methods A total of 22 healthy subjects [aged 27·7 ± 1·8 years; body mass index (BMI) 24·1 ± 0·8 kg m−2] participated in the study. After a 4-h euglycaemic hyperinsulinaemia (55·9 ± 2·2 mU L−1) insulin infusion was stopped and baseline blood samples were taken. Glucose infusion at a decreasing rate continued for 120 min to maintain euglycaemia throughout the study. Results The free fatty acid (FFA) concentration at the 120-min time-point was associated with baseline alpha-1-acid glycoprotein (A1GP) (r = 0·57, P

Diminishing ultrafiltration and dialysis adequacy may limit the long-term use of peritoneal dialysis (PD). Inflammation may play a role in changes in peritoneal function. This study was designed to evaluate alterations in peritoneal function and soluble factors in dialysate during a 1-year follow-up period.A personal dialysis capacity test was performed at the start of the study and after 6 and 12 months in 20 patients in order to determine dialysis adequacy and membrane characteristics. Dialysate was collected during the test days for analyses of interleukin-6 (IL-6), soluble intercellular adhesion molecule-1, hyaluronan and cancer antigen 125 (CA125).There were no significant changes in dialysis adequacy or membrane characteristics during the 1-year follow-up period. The appearance rate of IL-6 in dialysate increased significantly (419.8+/-63.3 at the start, 784.1+/-136.4 after 6 months and 1149.3+/-252.2 ng/24 h after 12 months; p=0.006) during follow-up. Furthermore, the appearance rate of CA125 increased throughout the study in patients using icodextrin, but decreased slightly in patients using only conventional dialysis solutions.There were no major changes in dialysis adequacy or membrane characteristics during the follow-up period, but increased IL-6 in dialysate may reflect peritoneal inflammation, which may lead to long-term alterations in the peritoneal membrane. Icodextrin may have a preventive effect on the longevity of the peritoneal membrane.

Aim: It has been shown that atherosclerosis is an inflammatory disease. Recent data suggest that inflammation precedes type 2 diabetes. Hence, we wanted to study the interrelationship between IL-6, insulin sensitivity, lipids and numerous acute-phase proteins. Methods: Twenty-one healthy individuals [16 males/5 females, age 27.9 ± 1.8 years, body mass index (BMI) 24.1 ± 0.8 kg/m2] participated in the study. Each patient went through a 4-h hyperinsulinaemic (40 mU/m2/min) euglycaemic clamp and 4-h saline infusion. Blood samples were taken before and at the end of the infusions. Results: Plasma interleukin (IL)-6 concentration correlated inversely with insulin sensitivity (M-value) (r = −0.49, p

Cytomegalovirus (CMV) is a suggested risk factor for chronic allograft nephropathy, and transforming growth factor-beta (TGF-beta) is a key fibrogenic molecule in this process. CMV has been shown to induce the expression of TGF-beta and several cytokines. We analyzed the impact of CMV on urinary excretion of TGF-beta, ICAM-1, TNF-alpha and correlated findings with biopsy histology.Urine samples from 46 renal transplant recipients were available for the study. Urine samples were taken when CMV infection was suspected, or for controlling of proteinuria or bacteriuria.CMV was diagnosed by antigenemia and viral cultures. Patients with previous CMV infection were excluded from the analysis. Urine samples were analyzed by ELISA-method to detect the levels of TNF-alpha, ICAM-1 and TGF-beta(1). Banff '97 criteria were used for scoring of protocol biopsies taken 6 months after transplantation.At the time of the urine collection, 13/46 patients had CMV infection. Eight patients with no CMV infection were used as controls. TGF-beta(1) was significantly increased in the CMV group (samples taken mean 137+/-79 days post-transplantation) compared to controls (samples 139+/-64 days post-transplantation) (51.1+/-28.0 vs. 13.3+/-6.7 ng/mmol crea, p0.001). No differences in the levels of other molecules were recorded. In the biopsies, interstitial fibrosis was significantly increased in the CMV group compared to controls.Urinary excretion of TGF-beta(1) was increased in patients during CMV infection. This was associated with increased fibrosis in the biopsies.

Membranous glomerulonephritis (MGN) has a variable clinical course, and factors that determine the prognosis are unknown. Our previous study suggested that urinary excretion of vascular endothelial growth factor (VEGF) is decreased in active MGN, but normalizes in remission. In the present study, VEGF protein and messenger RNA (mRNA) expression were investigated in this disease.Twelve patients with clinically active and/or progressive MGN were studied by using urinary assays for VEGF and soluble VEGF receptor-1 (sVEGF-R1), semiquantitative scoring of serial renal biopsy specimens by using immunohistochemical staining for VEGF protein, and in situ hybridization for VEGF mRNA. Results were compared with healthy controls and normal parts of nephrectomized kidneys.Urinary VEGF excretion was decreased significantly (P0.001 versus controls) in MGN, but there was no difference in sVEGF-R1 excretion compared with healthy subjects. VEGF protein expression was diminished significantly in glomerular podocytes (P = 0.008), as well as in extraglomerular small arteries (P = 0.03) and arterioles (P = 0.008) in MGN. Total kidney VEGF score (the sum of scores of individual kidney compartments) also was decreased in MGN (P0.05) and remained low in repeated biopsies. Expression of VEGF mRNA localized predominantly to podocytes in normal kidneys was greatly reduced in MGN.Clinically active MGN is associated with diminished expression of VEGF protein and mRNA, mainly in podocytes, and expression remains depressed in persistently active and/or progressive disease. This is reflected by decreased urinary VEGF excretion. These findings point to potentially reversible podocyte injury and, together with our previous study, suggest that VEGF may have a protective role during the evolution of MGN.

Aim: Since the elevated concentration of serum C-reactive protein (CRP) is a sensitive indicator of underlying inflammation, we investigated the association between serum CRP during the initial 6 post-transplantation months and histopathological changes in the 6-month protocol biopsies in 79 patients. We stained the biopsies for CRP and C3 to elucidate a possible role of CRP in renal injuries. Results: Forty patients showed no or minimal (Grade 0 - 1) tubular atrophy or interstitial fibrosis and 39 patients mild to moderate (Grade ≥ 2) chronic histopathological changes. The latter group had had higher concentration of CRP during the first 6 post-transplant months. Because the histopathological changes predict poor long-term prognosis, we followed-from 6th month onwards - 40 patients who had no or minimal histopathologic changes, and analyzed the association between CRP elevation and development of chronic allograft dysfunction. During this follow-up period (mean 51, range 14 - 72 months), 23 of 40 patients retained normal CRP level (Group A, mean CRP 1.12 mg/I), and 17 patients had elevated CRP concentrations (Group B, mean CRP 4.16 mg/l); 24-hour creatinine clearance improved or remained the same in all Group A patients, whereas it decreased in 7 of 17 (41%) of Group B patients (p < 0.001). In Group B patients, the annual change of creatinine clearance correlated inversely with the mean CRP concentration (r = -0.682, p < 0.01). Conclusion: Our results show that histological changes in 6-month biopsies were more prominent in patients with more transplantation-associated complications, infections and frequently higher CRP levels during the initial 6 post-transplant months than in those with lower CRP levels. During post-biopsy follow-up, we found low-grade systemic inflammation - measured as elevated CRP - to associate with impairment of graft function in patients with no or minimal histological findings in 6-month biopsies, and permanently low CRP to rule out chronic allograft dysfunction.

BACKGROUND Interstitial fibrosis in the protocol biopsy specimens of transplanted kidneys is regarded as the most reliable predictor of future impaired renal function. Type I and III collagens are the main components of renal fibrosis. During the synthesis and deposition of type III collagen, an amino-terminal propeptide (PIIINP) of a molecular weight of 44 kDa is degraded from the collagen and secreted into surroundings. Increased circulating PIIINP has been shown to reflect ongoing fibrotic processes. METHODS The extent of interstitial fibrosis in 6-month protocol biopsy specimens was recorded, and the urinary excretion of PIIINP in 24-hr urine specimens was measured in 79 graft patients. We also measured the urinary excretion of transforming growth factor (TGF)-beta 1, alpha(1)-microglobulin (alpha(1)M), and albumin and recorded the changes in creatinine clearance during 0.5 to 6 (mean, 4.3) posttransplant follow-up years. RESULTS The urinary excretion of PIIINP was significantly lower in patients with no interstitial fibrosis compared with patients with mild or moderate interstitial fibrosis (P

The aim was to determine the association of tear fluid cytokine levels and post-PRK corneal haze evaluated by in vivo confocal microscopy. In addition, the possible association between subbasal neural regeneration and haze formation, or epithelial regeneration were investigated. Twenty eyes of 20 patients (16 women and four men, age 30.7 +/- 7.5 years, range 21-48 years) underwent a myopic PRK. The spherical equivalent (SE) of the intended correction was -4.7 +/- 1.5 D (range -2.75 to -9.00 D). ELISA-methods were used to assess tear fluid concentrations of TGF-beta1, PDGF-BB and TNF-alpha pre-operatively, and post-operatively on day 2 and at 3 months. Tear fluid flow in the collection capillary was recorded, and rates of cytokine release (= tear fluid flow-corrected concentrations) were calculated. In vivo confocal microscopy was performed at 3 months to evaluate the corneal morphology and to determine numerical haze estimate. There was wide interindividual variation between pre-operative and post-operative concentrations and rates of release of TGF-beta1, PDGF-BB and TNF-alpha. Subepithelial haze was observed in all corneas and the mean haze estimate was 506 +/- 401 U (100-1410 U). However, no association was found between tear fluid cytokine levels and post-PRK haze. Regenerating subbasal nerve plexus was found in 18 out of 20 corneas; in two corneas it was absent or could not be visualized due to subepithelial haze. The density of the subbasal nerve fiber bundles had a positive correlation with the epithelial thickness (Pearson correlation, r = 0.56, P = 0.011), but not with the haze estimate or the thickness of the haze area. At 3 months post-PRK, haze could be observed in all patients. The results suggest that tear fluid cytokine analysis, as measured, may not be suitable for screening the potential candidates for haze formation. We did not find any correlation between haze and regeneration of subbasal nerve plexus, but we demonstrated that the regeneration of subbasal nerve plexus might have significant influence on regulation of epithelial healing.

BACKGROUND/AIMS—The Fas-Fas ligand system is thought to be involved in stromal cell apoptosis after corneal wounding. The aim was to measure changes in human tear fluid levels of soluble Fas (sFas) and Fas ligand (sFasL) following myopic photorefractive keratectomy (PRK). METHODS—Tear samples of 59 patients were collected preoperatively, and 1 or 2 days after PRK. Tear fluid sFas or sFasL concentrations were determined using sandwich ELISAs. Subsequently, tear flow corrected concentrations (releases) were calculated to compensate for the postoperative tear hypersecretion. RESULTS—The preoperative tear fluid flows (TFF) were 6.4 (1.7) µl/min (mean (SEM)) in sFas group (n = 18), and 7.5 (1.5) µl/min in sFasL group (n = 39). Postoperatively TFFs increased to 37.9 (10.9) µl/min (p = 0.003) and 58.3 (7.0) µl/min (p = 0.000), respectively. The mean preoperative sFas concentration (24.4 (11.6) U/ml) decreased to 9.7 (4.1) U/ml (p = 0.001) postoperatively, and the mean sFasL concentration (299.1 (28.8) ng/l) to 118.7 (15.9) ng/l (p = 0.000). However, the release of both substances increased significantly: sFas from 87.3 (29.4) mU/min to 229.4 (82.9) mU/min (p = 0.002) and sFasL from 1620.6 (226.4) fg/min to 4777.1 (596.1) fg/min (p = 0.000). CONCLUSIONS—Both sFas and sFasL are normal constituents of human tears. Despite a decrease in concentrations related to reflex tears, the release of sFas and sFasL increases significantly after excimer laser photorefractive keratectomy, which suggests that they are involved in corneal healing after PRK in humans.

To test the hypothesis that interaction between genetic, immunological, clinical and metabolic risk factors influences the outcome of Type II (non-insulin-dependent) diabetes mellitus, we examined which of the above factors present at baseline were associated with mortality in 134 Type II diabetic patients followed for 9 years. Thirty-eight patients (29 %) died during the follow-up period; the majority of whom (68 %) died from cardiovascular disease. At baseline, the deceased patients had higher HbA1 c values (p = 0.002), higher LDL-triglycerides (p = 0.007), lower HDL-cholesterol (p = 0.007), higher non-esterified fatty acid (NEFA) concentrations (p = 0.014), and higher albumin excretion rate (p < 0.0001) than the patients who survived. In addition, the frequency of HLA-DR4 (21 vs 39 %, p = 0.048) and of parietal cell antibodies (5 vs 14 %, p = 0.016) were decreased in the deceased as compared to the living patients. Patients who died during follow-up also had more retinopathy (42 vs 16 %, p = 0.002), neuropathy (57 vs 23 %, p < 0.001), microalbuminuria (45 vs 6 %, p < 0.0001), coronary heart disease (50 vs 13 %, p < 0.0001), and peripheral vascular disease (27 vs 9 %, p = 0.005) at baseline than patients who survived. In a multiple logistic regression analysis macroangiopathy (p = 0.004), neuropathy (p = 0.007), HbA1 c (p = 0.018) and albumin excretion rate (p = 0.016) were independent risk factors for death. In patients free of cardiovascular disease at baseline, conventional risk factors such as LDL-cholesterol (p = 0.005) and age (p = 0.003) were associated with subsequent development of cardiovascular disease. In conclusion, in addition to coexisting macroangiopathy, increased albumin excretion rate, poor glycaemic control and neuropathy are risk factors for cardiovascular mortality in patients with Type II diabetes. The presence of HLA-DR4 and signs of autoimmunity may be associated with decreased risk of cardiovascular disease. [Diabetologia (1998) 41: 1253–1262]

Abstract The basic feature in the pathogenesis of abdominal aortic aneurysm (AAA) is the degradation of extracellular matrix components. This process is induced partly by cytokines secreted from inflammatory and mesenchymal cells. Circulating levels of inflammatory cytokines were studied in AAA patients and compared with subjects suffering from atherosclerotic disease only. Furthermore, the predictive value of cytokine concentrations was evaluated for aneurysm expansion rate. Circulating levels of interleukin 1β (IL-1β), interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) were measured in 50 AAA patients (40 men, 10 women), 42 patients with coronary heart disease (CHD) (23 men, 19 women), and 38 controls whose angiogram was normal (17 men, 21 women). No differences in cytokine concentrations were found between the CHD patients and the controls. AAA disease was found to be associated with significantly higher IL- 1β and IL-6 concentrations in both male patients (median concentrations of 19.40 pmol/L and 6.45 pmol/L, respectively) and female patients (19.26 pmol/L and 7.99 pmol/L) than in either the CHD patients or the controls ( P P P P R =.37, P R =−.42, P

To measure the pre- and postoperative tear fluid platelet-derived growth factor-BB (PDGF-BB) concentrations of patients undergoing excimer laser photorefractive keratectomy (PRK).Tear fluid samples from PRK patients were collected with scaled microcapillary tubes preoperatively (Day 0, N = 23), on the second (Day 2, N = 24) and on the seventh (Day 7, N = 14) postoperative days. The PDGF-BB concentration was measured using a sandwich enzyme immunoassay, and the PDGF-BB release was calculated by multiplying the concentration by the tear fluid flow in the collection capillary.The mean tear fluid flow in the capillary was 17.4 microliters/ min (range 1.4-55.0) on Day 0, 62.6 microliters/min (4.3-125.0, p = 0.0000) on Day 2, and 15.5 microliters/min (1.2-50.0, NS) on Day 7. Seventeen percent (4/23) of the Day 0 samples contained detectable concentrations of PDGF-BB (range 95-1330 ng/l). On Day 2 all (24/24) tear samples showed significantly increased concentrations (mean 507 ng/l, range 45-2280 ng, p = 0.0001) and releases (mean 27.6 pg/min, range 1.0-98.0; p = 0.0000). On Day 7 PDGF-B was still measurable in 9/14 samples (mean concentration 194 ng/l, range5-925 ng/l, and mean release 1.9 pg/min, range 0.04-6.8 pg/min).PDGF-BB is an inducible component of tear fluid. In spite of hypersecretion caused by the corneal wound, PDGF-BB concentrations raised significantly during the initial wound healing. Increased PDGF-BB release during the two postoperative days following PRK suggests a role in corneal wound healing.

Neutral endopeptidase (EC 3.4.24.11; NEP), originally isolated from renal tubular brush border, is a cell surface peptidase identical to the CD10 antigen (or CALLA; common acute lymphoblastic leukemia antigen) in lymphoid cells. We studied the serum NEP levels daily after transplantation (Tx) in 19 renal allograft recipients. The NEP activity was determined with a two-step enzymatic assay utilizing a fluorogenic substrate (Suc-Ala-Ala-Phe-AMC; see text) and related to clinical signs of graft rejection, to signs of immunoactivation in transplant fine-needle aspiration biopsy (FNAB) specimens, to renal function, and to serum levels of C-reactive protein. The serum NEP levels remained normal (peak level 10.3 +/- 1.8 micrograms/l on days 6-9 after Tx, initial level after Tx 7.3 +/- 1.4 micrograms/1 on day 2; mean values +/- SEM) in patients who neither showed clinical signs of rejection nor had findings of immunoactivation in FNAB samples. On the contrary, the serum NEP levels rose clearly in patients developing acute rejection verified clinically and in FNAB samples (peak value 90.4 +/- 18.7 micrograms/l on days 6-9 post-Tx; p < 0.001 compared with patients without sings of immunoactivation) and even in patients having immunoactivation in FNAB without clinical evidence of rejection (108.2 +/- 22.4 micrograms/l, p < 0.001). Serum NEP peak appeared 2-3 days before clinical diagnosis of rejection and a positive findings in FNAB samples. Serum NEP increments did not correlate with changes in serum creatinine, delayed onset of renal excretory function, blood leukocyte count, C-reactive protein level, or infections. Thus, the serum NEP activity was shown to increase after renal allotransplantation associated with early phases of immunoactivation and development of acute graft rejection. Because of the limited number of patients studied, the clinical implications of these preliminary observations for kidney transplant monitoring clearly need confirmation in larger studies.

Microalbuminuria has recently been associated with insulin resistance in both insulin-dependent and non-insulin-dependent (NIDDM) diabetes mellitus. To establish whether microalbuminuria in non-diabetic subjects as well is associated with insulin resistance and associated abnormalities in glucose and lipid metabolism, oral glucose tolerance tests were performed with measurement of urinary albumin excretion rate, lipids and lipoproteins in 582 male non-diabetic first-degree relatives of patients with NIDDM. In addition, insulin sensitivity was assessed in 20 of these subjects with the euglycaemic hyperinsulinaemic clamp technique. Abnormal albumin excretion rate (AER), defined as AER 15-200 micrograms/min, was associated with higher systolic blood pressure (p0.05), higher fasting glucose values (p0.05), lower HDL-cholesterol (p0.05) and lower apolipoprotein A-I (p0.05) concentrations than observed in subjects with normal AER. The rate of glucose metabolism was lower in subjects with abnormal compared to subjects with normal albumin excretion rate (38.0 +/- 2.8 vs 47.3 +/- 2.4 mumol.kg lean body mass-1.min-1; p = 0.028). This difference was almost completely accounted for by a reduction in non-oxidative glucose metabolism (17.7 +/- 1.9 vs 27.4 +/- 2.7 mumol.kg lean body mass-1.min-1; p = 0.010), which correlated inversely with the AER (r = -0.543; p = 0.013). These results suggest that in non-diabetic individuals genetically predisposed to NIDDM, abnormal AER is associated with insulin resistance and abnormalities in glucose and lipid metabolism.

Thirty-five children with acute lymphoblastic leukemia were monitored weekly during the first 12 weeks of chemotherapy. The transferrin receptor (TfR) concentration was 2.8 ± 0.2 mg/I (mean ± S.E.M.) at diagnosis, decreased up to 3 weeks, and then increased reaching a maximal level at 8 weeks. The mean values for reticulocyte counts followed a similar pattern. In contrast, serum erythropoietin and ferritin levels were generally high. Those patients whose erythropoiesis was more accelerated had higher serum TfR concentrations. We conclude that among these patients the TfR level reflected the rate of erythropoiesis and was independent of the level of erythropoietin.

Idiopathic membranous glomerulonephritis (iMGN) has previously been shown to be associated with urinary excretion of terminal complement complexes while increased urinary levels of cytokines have been reported in mesangial proliferative glomerulonephritis. In the present cross-sectional study urinary excretion of IL-1 beta, TNF-alpha, IL-6, and soluble C5b-9 (SC5b-9) was examined for 23 patients with iMGN, 16 patients with diabetic nephropathy (DNP), and 17 healthy subjects. IL-1 beta excretion (pg/mg crea) was significantly higher in iMGN patients (375, range 162-11,000) than in DNP patients (39, range 22-59, P < 0.001) or healthy controls (151, range 23-481, P < 0.001). TNF-alpha excretion rate (pg/mg crea) was clearly higher (38, range 21-700) in iMGN patients than in DNP patients (14, range 8-52, P < 0.001) or healthy subjects (11, range 7-26, P < 0.001). Median IL-6 excretion (pg/mg crea) was only marginally higher in iMGN patients (73, range 0-850) than in healthy subjects (64, range 3-158, P = 0.02) but significantly higher than in DNP patients (29, range 17-47, P < 0.001). No significant correlation with corresponding serum values was observed for urinary IL-6 or TNF-alpha excretion. Urinary IL-1 beta and TNF-alpha correlated with decreased renal function. Five of 23 patients showed progression of iMGN over a follow-up of 6 months. The excretion of all cytokines, TNF-alpha in particular, was significantly higher in patients with a progressive disease than in the other patients.(ABSTRACT TRUNCATED AT 250 WORDS)

We performed a 24-week open clinical study in which 12 patients with rheumatoid arthritis (RA) and anemia (mean hemoglobin (Hb) value 102 g/l, range 90–109 g/l) were treated with recombinant human erythropoietin (rHuEPO). rHuEPO was given as a subcutaneous injection twice weekly with an initial dose of 300 U/kg/week. Nine of the II patients who completed the study responded with an increase in Hb value of 15 g/l or more within 3 to 17 weeks. Three months after treatment the Hb levels were significantly lower than the highest Hb levels (p < 0.0001). There was an inverse correlation between the response rate md the mean scrum concentrations of C-reactive protein and serum amyloid A protein (p

Serum immunoreactive interleukin-6 (ir-IL-6) concentration was measured by radioimmunoassay in nine patients with chronic lymphocytic leukaemia (CLL), 16 patients with multiple myeloma (MM), 12 patients with monoclonal gammapathy of undetermined significance (MGUS), 22 patients with primary Sjogren's syndrome (SS), and in 32 control subjects. Measurable quantities of ir-IL-6 were detected in every sample studied. Patients with MGUS and SS had significantly higher serum ir-IL-6 (mean +/- SD) concentrations (337 +/- 92 ng l-1 and 299 +/- 100 ng l-1, respectively) than controls (92 +/- 77 ng l-1) and patients with CLL and MM (120 +/- 32 ng l-1 and 113 +/- 58 ng l-1, respectively). Longitudinal studies of ir-IL-6 concentration in a few patients with MM showed a decrease before remission and an increase prior to relapse, but no consistent pattern was detected. In conclusion, we found that serum ir-IL-6 levels are higher in patients with benign hypergammaglobulinaemic states than in patients with malignant neoplastic B-cell disorders.

In order to study the role of interleukin-6 (IL-6) in inflammatory disease we monitored plasma levels of IL-6 and acute phase proteins such as C-reactive protein (CRP) and renin substrate (RS) in patients with reactive arthritis (ReA), rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE). Venous plasma samples were collected: (1) during the acute phase or exacerbation of the disease, and (2) several months latter during convalescence. Increased mean [95% confidence intervals (CI)] levels of plasma IL-6 were observed in patients with ReA both in the acute phase and later, 229 (177 to 280) ng/l and 197 (134 to 260) ng/l respectively (P less than 0.001 as compared to controls). The corresponding plasma IL-6 levels in RA patients were 283 (223 to 340) ng/l and 183 (151 to 226) ng/l, respectively (P less than 0.001 as compared to controls). Plasma IL-6 levels in SLE patients were not increased. Plasma RS levels were increased in all patient groups, but no significant correlation to IL-6 or CRP levels was observed, whereas plasma IL-6 and CRP levels showed a positive correlation in ReA and RA patients.

Tumor necrosis factor-alpha (TNF alpha) concentrations were measured by radioimmunoassay in sera of 118 children (median age, 1.7 years; range, 2 months-15 years) hospitalized for acute lower respiratory tract infection (ALRI). Both viral and bacterial ALRI were associated with elevated concentrations of TNF alpha. Concentrations greater than 40 ng/l were seen in children with bacterial or mixed ALRI in 64% and with viral ALRI in 50% of cases. Elevated concentrations were associated with longer duration of fever before admission (P less than .05) and with a higher serum C-reactive protein concentration (P less than .05). There were no significant differences in TNF alpha concentrations between gram-positive and gram-negative infections, nor was there an association with clinical severity of ALRI. TNF alpha concentrations decreased in most patients to normal within 5 days of hospitalization, irrespective of the etiology of the infection.

This study examined the connection between serum tumor necrosis factor (TNF) concentration and the development of cachexia in 12 children with acute lymphoblastic leukemia (ALL). The changes in muscle thickness were used as criteria for malnutrition, estimated by an ultrasound method during the 16 weeks of chemotherapy subsequent to diagnosis. Serum TNF concentrations were elevated at diagnosis and gradually decreased toward the reference limits by week 16. There was no correlation between TNF and muscle thickness. The results were also compared to those obtained from 8 children with other malignancies in whom the mean relative weight remained below normal whereas in those with ALL it gradually increased to + 15%. Thus, we found no evidence of the association between elevated serum TNF concentrations and cachexia in man.

Using commercially available antigens, enzyme-linked immunosorbent assays (ELISAs) were set up to demonstrate antibodies of IgG class against Sm and SS-A. Anti-Sm antibodies were demonstrated in 40% of patients with systemic lupus erythematosus (SLE), in 12% of patients with Sjogren's syndrome, in 6% of patients with rheumatoid arthritis (RA) and in 12% of patients with miscellaneous rheumatic disorders. Anti-SS-A antibodies were seen in 63% of the SLE patients, in 37% of the patients with Sjogren's syndrome and in 23% of the patients with RA. In the patients with SLE, high levels of anti-Sm antibodies were related to the presence of Raynaud's phenomenon, whereas patients with a malar rash tended to have high levels of anti-SS-A antibodies. In 17 SLE patients followed over a period of time a correlation was seen between the levels of the anti-Sm antibodies and the disease activity. We concluded that it is useful to include ELISAs for the demonstration of anti-Sm and anti-SS-A antibodies in determining the serological profile and in the follow-up of patients with SLE.

The effects of hemodialysis (HD) on the levels of serum amyloid A (SAA), C-reactive protein (CRP) and interleukin-1 beta (IL-1 beta) were studied in 8 patients. Bicarbonate dialysate was used exclusively, and three different membranes, Cuprophan (CU), cellulose acetate (CA), and polymethylmetachrylate (PMMA) were compared. The SAA levels increased significantly with each membrane. With CU, they rose from 4.0 +/- 2.0 (mg/l, mean +/- SEM) to 9.6 +/- 2.8 at 60 min and to 15.0 +/- 4.9 at 240 min. The values with CA were 3.8 +/- 2.1, 15.3 +/- 5.6, and 23.8 +/- 3.9; and with PMMA 2.4 +/- 1.3, 12.1 +/- 5.6, and 12.1 +/- 5.9, respectively. The alterations of SAA neither correlated with the weight loss nor the increase of serum albumin during dialysis. The CRP values showed insignificant changes. The IL-1 beta levels rose with CU from 87 +/-18 (ng/l) to 155 +/- 33 at 60 min and to 172 +/- 47 at 240 min. With CA, the values were 67 +/- 14, 198 +/- 46, and 121 +/- 23, and with PMMA 63 +/- 13, 246 +/- 93, and 211 +/- 86, respectively. These results did not correlate with the effects of the membranes on complement activation. It is concluded that the release of cytokines during HD apparently leads to a rapid synthesis of acute-phase proteins as a sign of inflammation. Thus, SAA may be used as one indicator of the biocompatibility of HD treatment.