Effect of hypoxia/reoxygenation on the cytokine-induced production of nitric oxide and superoxide anion in cultured osteoarthritic synoviocytes.

OBJECTIVE: Hypoxia/reoxygenation (H/R) is an important feature in the osteoarthritis (OA) physiopathology. Nitric oxide (NO) is a significant proinflammatory mediator in the inflamed synovium. The purpose of this study was to investigate the effects of H/R on inducible NO synthase (iNOS) activity and expression in OA synoviocytes. In addition we studied the relationship between nitrosative stress and NADPH oxidase (NOX) in such conditions. METHODS: Human cultured synoviocytes from OA patients were treated for 24 h with interleukin 1-ß (IL-1ß), tumour necrosis factor α (TNF-α) or neither; for the last 6 h, they were submitted to either normoxia or three periods of 1-h of hypoxia followed by 1-h of reoxygenation. ·NO metabolism (iNOS expression, nitrite and peroxynitrite measurements) was investigated. Furthermore, superoxide anion O2(·-) production, NOX subunit expression and nitrosylation were also assessed. RESULTS: iNOS expression and nitrite (but not peroxynitrite) production were ~0.20 to ~0.12 nmol min(-1) mg proteins(-1) (P < 0.05), while NOXs' subunit expression and p47-phox phosphorylation were increased. NOXs and p47-phox were dramatically nitrosylated under H/R conditions (P < 0.05 vs normoxia). Using NOS inhibitors under H/R conditions, p47-phox nitrosylation was prevented and O2(·-) production was restored at normoxic levels (0.21 nmol min(-1) mg of proteins(-1)). CONCLUSIONS: Our results provide evidence for an up-regulation of iNOS activity in OA synoviocytes under H/R conditions, associated to a down-regulation of NOX activity through nitrosylation. These findings highlight the importance of radical production to OA pathogenesis, and appraise the metabolic modifications of synovial cells under hypoxia.

[Evaluation of analytical and diagnostic performances of the anti-cyclic citrullinated peptides (anti-CCP) automated assay on Elecsys analyzer]. / Dosage automatisé des anticorps anti-peptides cycliques citrullinés (anti-CCP) sur l'analyseur Elecsys: validation analytique et valeur diagnostique.

Anti-cyclic citrullinated peptides (anti-CCP) are highly characteristics of rheumatoid arthritis (RA). Since 2006, anti-CCP assays have been included in both French and European recommendations. We have evaluated the analytical and clinical performances of the anti-CCP assay on the Elecsys analyzer (Roche Diagnostics). Two plasma pools (target values: 17.2 and 363.0 U/mL) and two quality controls (target values: 24.5 and 157.0 U/mL) were tested; we also analyzed three hundred plasma samples from healthy subjects (n = 86) and diseased patients (presenting with RA, non rheumatoid disorders, or undifferentiated arthritis: n = 214). Analytical performances (intra- and inter-assay precisions) and clinical performances (ROC analysis and method comparison) were evaluated. Elecsys assay was compared to Immunoscan RA(R) assay using contingency tables. Intra- and inter-assay precisions showed coefficients of variation less than 5%. ROC analysis showed an area under the curve at 0.886. Considering the value of 17 U/mL as the optimal cut-off, we found sensibility and specificity at 75% and 95%, respectively. Comparison of the Elecsys anti-CCP assay with the Immunoscan RA(R) assay showed an overall agreement of 98,3%. We conclude that the the Elecsys anti-CCP assay displayed a high precision and clinical performances comparable to that of the efficient anti-CCP assay Immunoscan RA(R).

N-Terminal pro B-type natriuretic peptide testing for short-term prognosis in breathless older adults.

BACKGROUND: Amino-terminal pro-brain natriuretic peptide (NT-proBNP) is useful for the triage of patients with dyspnea. Our aim was to determine whether NT-proBNP levels could predict in-hospital outcome in breathless elderly patients. METHODS: At admission, NT-proBNP plasma concentrations were determined in 324 dyspneic patients aged 75 years and older. The association between NT-proBNP values and in-hospital mortality was assessed. RESULTS: Median NT-proBNP concentrations were not different in deceased patients (n = 43, 13%) compared to that of survivors (n = 281, 87%) (4354 vs 2499 pg/mL, respectively; P = .06). To predict in-hospital mortality, the optimum threshold of NT-proBNP was 3855 pg/mL, as defined by the receiver operating characteristic (ROC) curve, with a nonsignificant area under the ROC curve of 0.59. Mortality was significantly higher in patients (n = 139) with NT-proBNP levels 3855 pg/mL or higher (17.9% vs 9.7%, P = .045). After multivariate analysis, NT-proBNP level 3855 pg/mL or higher at admission was predictive of mortality (odds ratio, 2.41; 95% confidence interval, 1.02-5.68; P = .04). CONCLUSION: NT-proBNP higher than 3855 pg/mL is associated with in-hospital mortality in patients aged 75 years and older admitted for dyspnea.

[The N-terminal pro-brain natriuretic peptide (NT-proBNP) assay with the Stratus CS analyzer]. / Evaluation des performances analytiques du dosage du NT-proBNP sur analyseur semi-automatisé Stratus CS.

BACKGROUND: NT-proBNP is an efficient biomarker for the evaluation, management and prognosis of patients with heart failure. METHODS: We evaluated the analytical performance of the NT-proBNP immunoenzymatic assay with the Stratus CS semi-automated analyzer in two hospital laboratories. The characteristics assessed included imprecision, functional sensitivity, linearity/recovery, interferences study, high-dose hook effect and a comparison of Acute Care(TM) pPBNP (on Stratus)CS) versus PBNP (on Dimension HM) results on patient heparinized plasma samples. RESULTS: Total imprecision reached < 5% coefficient of variation at NT-proBNP concentrations of 186-19,649 ng/L; recovery values for diluted samples were between 89.0 and 110.0 %; functional sensitivity reached 21 ng/L; there was no high-dose hook effect at concentrations up to 400,000 ng/L; hemaoglobin affected negatively but <10% the NT-proBNP assay, while bilirubin and triglycerides did not affected it more than 5%; Stratus CS results were strongly correlated with Dimension HM results (R(2)=1,00). CONCLUSION: The Stratus CS Acute Care pPBNP assay demonstrated excellent analytical performance which agreed with the Dimension HM PBNP assay. This analyzer is therefore suitable for use by low NT-proBNP test volume laboratories, and also by Emergency departments and Intensive care units.

Serum protein oxidation in patients with rheumatoid arthritis and effects of infliximab therapy.

OBJECTIVE: To examine protein oxidation in rheumatoid arthritis (RA) and evaluate its evolution after infliximab therapy in a subgroup of patients. METHODS: Seventy-one consecutive patients with RA were included. Among them, 30 patients refractory to conventional therapy were treated with infliximab. Serum markers of oxidative stress were determined at baseline and before the infusions of infliximab at weeks 6 and 30. Baseline values were compared with those in 30 healthy volunteers. RESULTS: Mean levels of serum carbonyl groups were significantly higher in RA patients than in controls (1.29+/-0.76 versus 0.58+/-0.39 nmol/mg of protein, p<0.0001), whereas thiol levels were found to be lower (238.3+/-61.6 versus 316.5+/-54.8 micromol/L, p<0.0001). Thiol levels inversely correlated with the disease activity score (r=-0.42, p=0.004), and with CRP values (r=-0.45, p=0.001). Immunoblots showed that albumin and heavy chain immunoglobulin were oxidized more markedly than in healthy volunteers. Significantly lower levels of thiol groups were detected in patients with refractory RA disease (208.9+/-66.8 versus 264.2+/-43.0 micromol/L, p<0.0004) but concentrations of carbonyl groups were similar. Short-term treatment with infliximab significantly decreased carbonyl groups (0.97+/-0.47 nmol/mg protein, p=0.02) and increased thiol (231.2+/-48.7 micromol/L, p=0.02) levels. CONCLUSION: Our results highlight free radical protein damage in RA and a link with inflammation, as underlined by the beneficial effects of infliximab.

Modulation of insulin action on 2-deoxyglucose uptake by chloroquine in chick embryo fibroblasts.

Blazar et al. recently found that chloroquine therapy decreased intravenous insulin requirements in a case of extreme insulin resistance. However, no relationship has been shown to exist between insulin degradation and the stimulation of glucose uptake. In this study we investigate the action of insulin on glucose uptake by the ability of this hormone to stimulate 2-deoxyglucose. The effect on alpha-aminoisobutyrate uptake, which is known to be insulin sensitive, is also investigated. Cell-associated 125I-labeled insulin and trichloroacetic acid-soluble and -precipitable substances were measured in parallel. Chloroquine increased insulin-stimulated uptake of 2-deoxyglucose and alpha-aminoisobutyrate. Three hours were required for this effect to appear, and it did not depend on DNA synthesis. Chloroquine also increased cell-associated insulin and slightly decreased the percentage of trichloroacetic acid-soluble products. Methylamine affected neither nutrient uptake processes nor insulin binding and degradation; however, it did abolish the effect of chloroquine on these parameters. These data suggest that in chick embryo fibroblasts a relationship may exist between the increase in undegraded cell-associated insulin and the ability of the hormone to stimulate sugar and amino acid uptake.

Influence of renal function on N-terminal pro-brain natriuretic peptide (NT-proBNP) in patients admitted for dyspnoea in the Emergency Department: comparison with brain natriuretic peptide (BNP).

BACKGROUND: Renal dysfunction influences the optimum brain natriuretic peptide (BNP) threshold for a diagnosis of cardiac-related dyspnoea, but this has not been demonstrated for N-terminal pro-brain natriuretic peptide (NT-proBNP). We studied the influence of renal function on NT proBNP and BNP concentrations in dyspnoeic patients admitted by night to the Emergency Department (ED). METHODS: NT-proBNP, BNP, and creatinine levels were measured in blood samples collected routinely from 381 patients; estimated glomerular filtration rate (eGFR) was calculated. RESULTS: Cardiac-related dyspnoea was found in 115 patients (30.2%). NT-proBNP and BNP values were elevated in patients with cardiac-related dyspnoea (6823+/-6569 vs. 2716+/-4838 pg/ml, and 642+/-329 vs. 243+/-267 pg/ml, p<0.0001, respectively). Log-transformed NT-proBNP and BNP values were correlated to eGFR values. Mean NT-proBNP and BNP values stratified by ED diagnosis increased in line with eGFR categories, but in each category both peptide concentrations remained elevated in cardiac-related dyspnoea when compared with non-cardiac-related dyspnoea (p<0.05). NT-proBNP (and BNP) cut-off points rose as a function of eGFR categories: from 1360 (and 290) pg/ml in patients with eGFR 60-89 ml/min/1.73 m2, to 6550 (and 515) pg/ml in patients with eGFR 15-29 ml/min/1.73 m2. CONCLUSION: Renal function influences the optimal cut-off points of NT-proBNP and BNP for the diagnosis of cardiac-related dyspnoea.

Effect of insulin on aminoisobutyric acid uptake by human non-rheumatoid and rheumatoid synovial cells.

The biological sensitivity of cultured non-rheumatoid human synovial cells (NRSCs) and rheumatoid synovial cells (RSCs) was examined in terms of the ability of insulin to stimulate the uptake of alpha-aminoisobutyrate (AIB). NRSCs, like numerous fibroblastic lines, were sensitive to physiological concentrations of the hormone: half-maximal stimulation was obtained with (4 X 10(-10) M) insulin, while maximum transport was found with a 60-90 min association time. On the contrary, although the basal transport was similar in RSCs, insulin was totally unable to accelerate AIB transport in these cells. Inflammatory processes lead to an insulin resistance which most likely involves a post-receptor step at the cellular level.

IL-1 beta, a strong mediator for glucose uptake by rheumatoid and non-rheumatoid cultured human synoviocytes.

Higher basal 2-deoxy-D-glucose uptake in rheumatoid synovial cells than in non-rheumatoid synovial cells, was found to be associated with an increased interleukin-1 beta (IL-1 beta) secretion (respectively 850 +/- 238 vs. 8.3 +/- 2.4 pg/24 h/10(5) cells, mean +/- S.E.M.). When exogenous human recombinant IL-1 beta was added to cultures, a marked stimulation of 2-deoxy-D-glucose uptake was performed by both human synovial cultured cells, in a time-dependent and dose-dependent manner (IL-1 beta 0-100 ng/ml). In non-rheumatoid synoviocytes, stimulation occurred 1-3 h following the addition of 1 ng/ml interleukin-1 beta and increased up to 24 hours (respectively +150% and +261.4% after 6 and 24 hours association time). Rheumatoid synovial cells were less sensitive to 1 ng/ml IL-1 beta (respectively +80% and +146.4%). IL-1 beta increased significantly the Vmax for 2-deoxy-D-glucose uptake by synovial cells, with no change in the Km. This effect was protein synthesis-dependent, and not secondary to prostaglandin E2 synthesis or cell growth. IL-1 beta possesses an important effect on glucose homeostasis in synovial cells, which could be indirect and/or regulated by the presence of natural inhibitors.

Insulin-like growth factor I/somatomedin C action on 2-deoxyglucose and alpha-amino isobutyrate uptake in chick embryo fibroblasts.

Maximum 125I-IGF-I/Sm-C total binding to chick embryo fibroblasts was 3% at +37 degrees C and decreased to less than 1% in presence of 2.8 X 10(-9) M unlabelled IGF-I/Sm-C. Insulin did not compete with IGF-I/Sm-C for the binding to cells. Biological action of IGF-I/Sm-C was evaluated on 2-deoxyglucose and alpha-aminoisobutyrate uptake. Results are compared with those obtained with insulin. Maximal peptide effects on the two transport processes were obtained at a 0.65 X 10(-7) M concentration and for a 120 minute association time, whereas cells were markedly less sensitive to insulin and time response curves were different. These results suggest that insulin action on nutrient uptake in chick embryo fibroblasts is not mediated by the binding of the hormone to IGF-I/Sm-C receptors.

Action of anti-inflammatory drugs on interleukin-1 beta-mediated glucose uptake by synoviocytes.

Synovial cell cultures prepared from samples taken from osteoarthritic and rheumatoid patients were treated with different anti-inflammatory agents (cortisol, indomethacin, ibuprofen and piroxicam) to determine their 'anti-interleukin-1 beta' action, using inhibition of interleukin-1 beta-mediated glucose uptake stimulation as a biological test. Confluent cells were treated for 24 h with different concentrations of these drugs (10(-5), 10(-6) and 10(-7) mol/l) to study their effect on the inflammation process. 6 h before glucose uptake studies, interleukin-1 beta (1 ng/ml) was added. Whereas non-steroid anti-inflammatory agents were inefficient, cortisol inhibited the action of interleukin-1 beta on glucose uptake. In osteoarthritic cells, cortisol, 10(-5) mol/l, reduced interleukin-1 beta-mediated glucose uptake by 27% after a 24-h incubation. In rheumatoid cells, stimulated 2-deoxy-D-glucose uptake was reduced by 40.6%. Results were similar when interleukin-1 beta and cortisol were added simultaneously, 6 h before glucose uptake was measured. This rapid effect of cortisol was protein synthesis-dependent (inhibited by cycloheximide). Cortisol decreased glucose uptake by synoviocytes by acting on basal and interleukin-1 beta-mediated glucose uptake. This effect was more pronounced in rheumatoid synovial cells. The inhibition of interleukin-1 beta-mediated glucose uptake could be proposed as a new model for studying the anti-interleukin-1 beta effects of anti-rheumatic drugs.

Variability in urinary excretion of bone resorption markers: limitations of a single determination in clinical practice.

In this study we assessed the within and between-subject variability of the concentrations of two urinary markers, free deoxypyridinoline (DPD) and C telopeptide (CTX-I), in healthy patients with the aim of setting reliable thresholds to enable physicians to take decisions about individual patients with confidence.Between-subject variability for the women was 25.4% for DPD and 38.2% for CTX-I, and for the men was 12.9% for DPD and 23.8% for CTX-I. The coefficients of variation were similar for daily, weekly and monthly determinations, giving means of 13.8 and 28.1% for DPD and CTX-I respectively. Critical difference (CD) was lower for DPD than for CTX-I (about 44 and 80% respectively). The number of samples required to determine the true mean with a CD at the 5% level was 29 for DPD and more than 113 for CTX-I.DPD was the least biologically variable. One determination was not sufficient to determine bone resorption status and a 44% decrease in DPD levels and an 80% decrease in CTX-I levels were required to demonstrate the efficacy of antiresorptive therapy in individual patients.

Stimulation of alpha-(methylamino) isobutyric acid uptake by interleukin-1 in human synovial cells. Involvement of a cAMP dependent pathway.

After one hour incubation with interleukin-1 beta (IL-1 beta), the uptake of alpha-(methylamino) isobutyric acid (MeAIB) by human osteoarthritic synovial cells appeared significantly increased. This effect, observed with 0.1 to 5 ng/ml of cytokine, was inhibited by cycloheximide, indicating that protein synthesis is involved. In addition, this effect seems mediated by a pertussis toxin-sensitive G protein. Finally, intracellular cAMP concentration measurements, the use of a phorbol ester, protein kinase inhibitors and forskolin+3-isobutyl-1-methylxantine (IBMX) provided evidence that a cAMP-dependent protein kinase is associated with interleukin-1 beta-mediated alpha-(methylamino) isobutyric acid uptake.

Is alpha-ketoisocaproyl-glutamine a suitable glutamine precursor to sustain fibroblast growth?

BACKGROUND: Glutamine is considered an essential nutrient for cellular growth. AIM: To test the suitability of alpha-ketoisocaproyl-Gln (Kic-Gln) as a new glutamine (Gln) precursor to sustain human fibroblast growth. METHODS: [3H] thymidine uptake into cellular DNA of human fibroblasts. Extracellular and intracellular amino acid patterns were determined with peptides and acylated compounds. RESULTS: L-alanyl-L-glutamine (used here as a recognized Gln precursor) promoted DNA synthesis, while N-acetyl-L-glutamine (used here as a negative control since it is known to be a poor Gln precursor) and alpha-ketoisocaproyl-glutamine had no effect. Alanyl-glutamine progressively gave rise to free glutamine in the growth medium. In contrast, glutamine supplied in acylated form was poorly available and did not appear in free form in the medium. In addition, only alanyl-glutamine increased intracellular glutamine and glutamate levels. In contrast, Kic-Gln was able to sustain net protein synthesis as judged by total protein content and reduced intracellular levels of most essential amino acids. CONCLUSION: Kic-Gln appears to be a poor extra-cellular precursor of Gln to sustain cell growth.

Type IV hyperlipoproteinemia in patients with algodystrophy.

The present work studies lipid metabolism in patients with algodystrophy (AD). A correct positive correlation (r = 0.75) between the triglyceride levels and low density lipoprotein (LDL)/very low density lipoprotein (VLDL) ratio and the VLDL increase observed by gel disk electrophoresis confirm that a type IV hyperlipoproteinemia is associated with AD. In contrast, the degree of high density lipoprotein (HDL) saturation in cholesterol (HDL-cholesterol/HDL-phospholipids) and the classical atherogenous index (cholesterol/HDL-cholesterol) were not modified. The decrease of plasma post-heparin lipolytic activities (PHLA) was not significant but further studies should be performed to correlate PHLA with a reduced activity of the adipose tissue lipoprotein lipase.

Fate of enterally administered ornithine in healthy animals: interactions with alpha-ketoglutarate.

In order to improve our understanding of the metabolic interactions between alpha-ketoglutarate (alpha KG) and ornithine (Orn), which constitute the two parts of ornithine alpha-ketoglutarate (OKG) used as an adjuvant in enteral nutrition, we have investigated the plasma appearance and tissue distribution (qualitative and quantitative) of enterally administered 14C-Orn and 14C-alpha KG in healthy mice and rats. The influence of unlabelled alpha KG or Orn on 14C-Orn or 14C-alpha KG metabolism, respectively, was also studied. Unlabelled alpha KG was able to reduce strongly the rate of intestinal absorption of 14C-Orn, whereas the inverse was not true. This alpha KG-induced loss in plasma radioactivity after a load of Orn was associated with a decrease of radioactivity in tissue with no modification of the qualitative distribution in organs. In this study, a direct interaction between alpha KG and Orn was demonstrated at the intestinal level. The mechanisms involved in this phenomenon probably involve the regulation of metabolic conversions among alpha KG, Glu, pyrroline-5-carboxylate, and Orn. This is of importance in the therapeutic use of ornithine salts in clinical nutrition.

Action of enterally administered ornithine alpha-ketoglutarate on protein breakdown in skeletal muscle and liver of the burned rat.

Several studies concerning burn patients have shown that supplementation of enteral nutrition with ornithine alpha-ketoglutarate (OKG) favorably modifies protein metabolism. Therefore, the effect of OKG administration on muscular and hepatic protein catabolism was evaluated in burned rats. Four groups of six rats were used. Two groups were scalded by immersion of the dorsum in water at 90 degrees C for 10 seconds and then starved for 24 hours. Controlled enteral nutrition was then administered in three boluses daily (Osmolite, 210 kcal/kg/d, 1.2 g N/kg/d); one group was supplemented with OKG (5 g/kg/d, ie, 0.68 g N/kg/d), while the other group received an equivalent amount of nitrogen in the form of glycine. One group of healthy control rats received Osmolite supplemented with glycine and the last group was fed ad libitum. The animals were killed after 2 days of nutrition. Protein catabolism was assessed in vitro by measuring the amount of valine (liver catabolism) and phenylalanine (muscle catabolism) released into the incubation medium of isolated tissues. Tissular and serum glutamine were also assayed. Burn injury induced muscle hypercatabolism without affecting hepatic catabolism. The administration of OKG limited both muscle weight loss and muscle protein hypercatabolism and significantly improved the muscle glutamine pool. These results demonstrate the nitrogen-sparing effect of OKG in muscle in hypercatabolic states.

Ornithine alpha-ketoglutarate and glutamine supplementation during refeeding of food-deprived rats.

The aim of this study was to compare the efficiency of ornithine alpha-ketoglutarate (OKG) and glutamine supplementation in an experimental model of denutrition that provides well-characterized disturbances of amino acid patterns. Male Wistar rats (187 +/- 11 g; five in each group) were starved for 3 days and then refed for 7 days with an oral diet (192 kcal kg-1.day-1 and 2.25 g of nitrogen kg-1.day-1), supplemented with 0.19 g of nitrogen kg-1.day-1 in the form of OKG, glutamine, or casein (control group). Food deprivation induced a fall in most tissue amino acids, with the notable exception of muscle leucine and liver glutamate, which increased by 43% (p < .01), and 11% (p < .05), respectively. The main effect of OKG was seen in the viscera, with a normalization of most amino acid pools (including proline and branched-chain amino acids) in the small bowel and liver. The main effect of glutamine was observed in the muscle, with a normalization of the glutamine and leucine pools. We conclude that, in this model and with the doses used, OKG and glutamine act in different target tissues, ie, splanchnic areas and muscle, respectively.

Relationship between acid neutralization capacity of saliva and gastro-oesophageal reflux.

Saliva is an important factor in neutralization of oesophageal acid exposure, clinically manifested as gastrooesophageal reflux (GOR). The aim of this study is to compare the composition and the "capacity in acid neutralization" (CAN) of saliva in controls and patients suffering of GOR. We compared the composition of saliva from 56 patients who had symptoms of GOR with that of saliva from 20 healthy control subjects. After a standardized 24-hour period of pH-monitoring, 39 patients had normal pH reflux scores (normal acid score: NAc-GOR) and 17 had abnormal pH reflux scores (pathological acid score: PAc-GOR). Then, following a 10-h fast, total saliva was collected during ten minutes in all patients and in the healthy control subjects. The composition of the saliva samples was analysed and the titration curve was determined on 200 microliters aliquots by successive addition of 5 microliters volumes of 0.1 N Hcl. The GOR patients had significantly lower salivary concentrations of Na+ and of both free and bound sialic acids and had higher salivary concentrations of inorganic phosphates than the controls. These disorders were more marked in PAc-GOR patients. Initial pH was 7.43 +/- 0.43 in controls, 7.35 +/- 0.45 in NAc-GOR patients, and 6.91 +/- 0.53 in PAc-GOR patients. In the beginning of the titration curve, PAc-GOR patients were significantly different from NAc-GOR patients and from controls. Saliva of both groups of patients presented significant differences in the acidic portion of the titrations curves, at high volumes of added HCl. These data show that the composition of saliva was modified in patients with GOR disease compared to that of normal subjects. A difference in titration curves was also observed with a higher acidic buffering capacity in these GOR patients. The modifications in saliva composition suggest a role for inorganic phosophates in the acid neutralization capacity observed in GOR, perhaps linked with a adaptation to chronic acid exposure.

Fibroblast cultures in experimental disturbed metabolic conditions. effects of anti-inflammatory drugs.

Fibroblasts in normal metabolic conditions constitute a simple cellular model for the study of anti-inflammatory drugs in culture, but do not take into account the factors related to the acute inflammatory reaction. A new approach was tried in submitting the fibroblasts to the action of a mouse peritoneal exudate or selected activated macrophages. Disrupted peritoneal material was inoculated to cultures in a volume representing a number of cells corresponding to 25 or 50% of fibroblast concentration at day 0 of the experimental time. Three anti-inflammatory drugs were retained to study their effects on proliferation and metabolic parameters: hydrocortisone, indomethacin and a newly-described molecule, the methoxybenzalthiosemicarbazone (MBT). At day 0, 1, and 2, fibroblasts were counted and total cellular protein and cellular glycolytic activity were determined. 4.5 X 10(-5) M hydrocortisone and MBT protected the fibroblast from the antiproliferative action of peritoneal exudate whereas in experiments using activated macrophage homogenate, all drugs exerted a protective and stimulatory effect.