Fracture discrimination by combined bone mineral density (BMD) and microarchitectural texture analysis.

The use of bone mineral density (BMD) for fracture discrimination may be improved by considering bone microarchitecture. Texture parameters such as trabecular bone score (TBS) or mean Hurst parameter (H) could help to find women who are at high risk of fracture in the non-osteoporotic group. The purpose of this study was to combine BMD and microarchitectural texture parameters (spine TBS and calcaneus H) for the detection of osteoporotic fractures. Two hundred and fifty five women had a lumbar spine (LS), total hip (TH), and femoral neck (FN) DXA. Additionally, texture analyses were performed with TBS on spine DXA and with H on calcaneus radiographs. Seventy-nine women had prevalent fragility fractures. The association with fracture was evaluated by multivariate logistic regressions. The diagnostic value of each parameter alone and together was evaluated by odds ratios (OR). The area under curve (AUC) of the receiver operating characteristics (ROC) were assessed in models including BMD, H, and TBS. Women were also classified above and under the lowest tertile of H or TBS according to their BMD status. Women with prevalent fracture were older and had lower TBS, H, LS-BMD, and TH-BMD than women without fracture. Age-adjusted ORs were 1.66, 1.70, and 1.93 for LS, FN, and TH-BMD, respectively. Both TBS and H remained significantly associated with fracture after adjustment for age and TH-BMD: OR 2.07 [1.43; 3.05] and 1.47 [1.04; 2.11], respectively. The addition of texture parameters in the multivariate models didn't show a significant improvement of the ROC-AUC. However, women with normal or osteopenic BMD in the lowest range of TBS or H had significantly more fractures than women above the TBS or the H threshold. We have shown the potential interest of texture parameters such as TBS and H in addition to BMD to discriminate patients with or without osteoporotic fractures. However, their clinical added values should be evaluated relative to other risk factors.

[Urinary prostaglandins in the hepatorenal syndrome of cirrhotic patients: role of thromboxane A2 and an imbalance of precursor polyunsaturated fatty acids]. / Prostaglandines urinaires dans le syndrome hépatorénal du cirrhotique: rôle du thromboxane A2 et d'un déséquilibre des acides gras polyinsaturés précurseurs.

The hepatorenal syndrome is considered to be a functional renal failure due to active renal vasoconstriction. The purpose of this work was to study the urinary elimination of prostaglandins and the plasmatic polyunsaturated fatty acid precursors of prostaglandins. The urinary elimination of PGE2 was not significantly different in the groups of patients studied: controls, group I (193 +/- 42 ng/24 h), cirrhotic patients without ascites, group II (274 +/- 43 ng/24 h), cirrhotic patients with ascites, group III (269 +/- 41 ng/24 h). The urinary elimination of PGE1 and PGF2 alpha varied in the same way as PGE2. In cirrhotics with hepatorenal syndrome (group IV) the urinary elimination of vasodilating prostaglandins was greatly decreased (p less than 0.001); PGE2 (53 +/- 16 ng/24 h), PGE1 (65 +/- 11 ng/24 h). The urinary elimination of PGF2 alpha was also decreased (293 +/- 75 ng/24 h). On the other hand, the urinary elimination of thromboxane, a vasoconstrictor, increased progressively from group I (287 +/- 75 ng/24 h) to group IV (980 +/- 266 ng/24 h) (p less than 0.05). Plasmatic arachidonic acid was significantly decreased in group IV (5.0 +/- 0.6 p. 100) compared to group I (10.0 +/- 0.6 p. 100) (p less than 0.001), to group II (10.3 +/- 0.5 p. 100) (p less than 0.001), and to group III (8.5 +/- 0.7 p. 100) (p less than 0.05). In conclusion, in hepatorenal patients, urinary excretion of a vasoconstricting prostaglandin (thromboxane) is increased while urinary excretion of vasodilating prostaglandins is decreased. This decrease could be secondary to a lack of plasmatic arachidonic acid, precursor of prostaglandins.

Effects of dietary gamma-linolenate supplementation on serum lipids and platelet function in insulin-dependent diabetic patients.

In order to gain insight into the mechanism of platelet dysfunction in insulin-dependent diabetics we studied in 17 patients the influence of a 6-week period with a dietary supplement of gamma-linolenate: 2 g/d (group I, n = 8) or 500 mg/d (group II, n = 9). Serum lipids, plasma beta-thromboglobulin (beta TG), platelet aggregation in vitro, and TxB2 and PGE1 released from platelets during the aggregation process were measured. In group I, serum triglycerides fell from 1.57 +/- 0.28 mmol/l to 0.99 +/- 0.17 mmol/l (P less than 0.01) and cholesterol fell from 5.85 +/- 0.55 mmol/l to 5.08 +/- 0.52 mmol/l (P less than 0.01). In group I plasma beta TG fell from 100.0 +/- 15.7 ng/ml to 73.7 +/- 12.1 ng/ml (P less than 0.025), while the sum of the percentages of the C18:3 omega 6 and its chain elongated (C20:3 omega 6) and desaturated (C20:4 omega 6) metabolites increased in serum triglycerides (P less than 0.05), cholesterol esters (P less than 0.02) and phospholipids (P less than 0.02). No changes were observed on the other parameters in either group. The results show that the lowering effects of gamma-linolenate on serum triglycerides, cholesterol and plasma beta TG occur only with daily intakes of 2 g. The changes in fatty acid composition of serum lipids suggest that the gamma-linolenate intake of 2 g may exert its beneficial effect through an increased incorporation of long-chain polyunsaturated fatty acids, but no firm conclusion can be drawn as membrane platelet fatty acid composition was not evaluated.

Long term treatment with theophylline increases malonyl dialdehyde plasma concentrations.

As theophylline is known to stimulate lipolysis and lipid mobilisation, plasma concentrations of malonyl dialdehyde were studied in asthmatic patients being treated with sustained release theophylline. Malonyl dialdehyde was assessed as malonyl dialdehyde like material (MDA-LM) by the thiobarbituric acid assay. In a kinetic study (eight patients) plasma MDA-LM concentrations were found to be significantly higher after 30 than after eight days' treatment or before treatment, but not significantly different after 90 than after 30 days. In a comparative study (29 patients and 29 healthy volunteers) the mean plasma MDA-LM concentration was shown to be very significantly higher in the asthmatic group. The results are probably explained by an effect of theophylline on lipid peroxidation, and the effect of long term treatment on plasma MDA-LM merits further investigation.

Plasma lipid fatty acids and platelet function in insulin-dependent diabetic patients.

Platelet function, estimated from plasma beta-thromboglobulin (beta-TG, ng/ml), is frequently altered in insulin-dependent diabetics (IDDs). As several factors may affect beta-TG, we studied respectively in 15 IDDs, the roles played by: (i) diabetic control evaluated from glycosylated haemoglobin (HbA1); (ii) plasma C-peptide and pancreatic glucagon; (iii) plasma lipids and the relative percentages of fatty acids in total plasma lipids. Plasma beta-TG did not correlate significantly with the first 3 parameters. However, beta-TG was correlated: (i) positively with plasma triglycerides (P less than 0.01), cholesterol (P less than 0.02), phospholipids (P less than 0.05) and total plasma lipids (P less than 0.01) and the percentage of oleic acid (C18 : 1 omega 9) in plasma lipids (P less than 0.01); (ii) negatively with the percentage of linoleic acid (C18 : 2 omega 6) in plasma lipids (P less than 0.02). No correlation was found between beta-TG and the percentages of the other saturated (C16 : 0, C18 : 0), monounsaturated (C16 : 1 omega 7) and polyunsaturated fatty acids (C18 : 3 omega 6, C20 : 3 omega 6 and C20 : 4 omega 6). The present results indicate that beta-TG in IDDs can be markedly improved by all dietary and therapeutic measures which lower plasma lipids and increase the percentage of the linoleic acid in the body.

Suppressor effect of prostaglandins on T colony formation.

This study evaluated the action of prostaglandins on T colony formation. A single step culture process was used which involved direct seeding of freshly isolated peripheral blood mononuclear cells (MC) in semi-solid agar culture medium containing phytohaemagglutinin (PHA). Suppression of endogenous production of prostaglandins with 10(-6) M indomethacin increased T colony formation by up to 100%. Similarly, addition of synthetic prostaglandin E (PGE) to the culture system demonstrated a dose-dependent reduction of T colony formation by PHA-stimulated non-adherent cells. The 50% inhibitory dose (ID 50) was 10(-7) M for PGE2 and 1.3 x 10(-7) M for PGE1. Prostaglandin F had no effect on T colony formation. The synthesis of PGE by adherent cells can be increased two- to three-fold in the presence of T colony promoting activity released by PHA-stimulated lymphocytes. We conclude that monocyte produced PGE is responsible for the suppressor effect exerted by these cells on T colony formation. The PGE inhibitory role is interpreted as a feedback mechanism, modulated by lymphokines released by PHA-activated lymphocytes.

Responses to interferon of LM cells growing continuously in the absence of serum and exogenous lipids.

Mouse fibroblast LM cells growing continuously in the absence of serum and exogenous lipids are proposed as a model for studying interferon action. Several of its activities, namely the inhibition of virus and cell growth, and the increased cytotoxicity of poly(rI) X poly(rC), are shown to be potentiated in such conditions. Since cells cultured under these conditions are devoid of precursors of prostaglandin biosynthesis, the data suggest that none of the three effects of interferon depends on active prostaglandin synthesis.

[Effect of the hormonal environment on the human oocyte and its later development in vitro]. / Influence de l'environnement hormonal de l'ovocyte humain sur son évolution ultérieure in vitro.

The biosynthesis of progesterone by the granulosa cells that come from follicles from which the oocyte has been fertilized and has divided in vitro is far superior to that from the cells coming from follicles in which the oocyte has failed to develop. This result is in accordance with the raised levels of progesterone and the influence of androgens on lower levels of oestrogens in follicular fluid that evolved favourably. In the same follicles the ratio of concentrations of prostaglandins PGE2 to PGF2 alpha is twice as high as in the liquid from other follicles.

[In vitro development of human ovocytes collected during spontaneous ovulation. Effect of the biochemical composition of the follicular fluids]. / Evolution in vitro d'ovocytes humains recueillis lors d'ovulations spontanées. Influence de la composition biochimique des liquides folliculaires.

Development of the in vitro cultured and fertilized ovocyte appears to be improved when progesterone and estrogen concentrations in the follicular fluid are above 2 000 and 1 000 ng/ml respectively, with a A/E ratio under 1 and a PGE/PGF ratio above 0.8. Such criteria are only infrequently fulfilled: once in seven cases. They were found in one of the two patients who had a pregnancy after in vitro fertilization during induced ovulation. Division of ovocytes from spontaneous ovulation was recorded in several instances in spite of a less propitious composition of the follicular fluid.

Functional assessment of alveolar macrophages: comparison of cells from asthmatics and normal subjects.

Alveolar macrophages (AM) were obtained by bronchoalveolar lavage (BAL) from seven healthy nonallergic, nonasthmatic donors, 15 patients with allergic bronchial asthma, and six patients with aspirin-sensitive asthma. AM were purified by adherence over 2 hr and cultured for an additional 24 hr. Functional assessment of viable cells was carried out for zymosan phagocytosis and for prostaglandin (PG) E2-PGF2 alpha and thromboxane (Tx) B2 release by resting and zymosan-stimulated AM. The eosinophil count in BAL fluid from allergic asthmatics was higher than that from control subjects (3.9% +/- 1.6% vs 0.4% +/- 0.3%, p less than 0.05) and still greater in BAL from patients with aspirin-sensitive asthma (21.7% +/- 9.0%, p less than 0.01). After the 24 hr of incubation, the AM viability was inversely correlated to the percentage of eosinophils in BAL fluid (r = -0.54, n = 21, p less than 0.02). Zymosan phagocytosis was significantly lower by viable cells from both allergic asthmatics and aspirin-sensitive patients as compared with cells from normal donors (p less than 0.05). Zymosan phagocytosis induced a twofold to threefold increase in the release of PGE2, PGF2 alpha, and TxB2 from AM of normal subjects (p less than 0.01) but only a onefold to twofold increase from AM of allergic asthmatic patients. The stimulated AM from aspirin-sensitive patients released smaller quantities of each product than AM from normal subjects or allergic asthmatic patients (p less than 0.05). We conclude that the viability and functional activity of AM are impaired in asthmatic patients and that these deficits correlate with the percent eosinophilia in the BAL; it is therefore suggested that they may be due to an interaction between eosinophils and AM in the bronchoalveolar lumen.

Effect of oestradiol 17 beta on prostaglandin biosynthesis by the uterus of ovariectomized rat and guinea pig.

In vitro prostaglandin biosynthesis by uteri of ovariectomized rats and guinea pigs treated or untreated with oestradiol 17 beta, administered subcutaneously, was measured by R.I.A. of PGF2 alpha and PGE2. Incubations with [1-14C] arachidonic acid were also performed and labelled metabolites were analyzed by TLC. The main metabolite in rats was 6 keto PGF1 alpha and in decreasing order of magnitude, PGF2 alpha and PGE2. In guinea pig PGF2 alpha was the main product. Ovariectomy in rats completely changed the pattern of synthetized prostanoids : PGI2 production was doubled when compared to cycling rats and PGE2 increased 10 fold. PGF2 alpha values were similar to the mean value measured during the cycle. OE2 treatment almost completely inhibited PGI2 synthesis and reduced PGE2 by half. Total PG synthesis in OE2 treated animals was decreased by 5 fold when compared to spayed rats. Endogenous PGF2 alpha synthesis was slightly stimulated. In the guinea pig OE2 treatment of ovariectomized animals increased the total synthesis from 50 per cent. PGF2 alpha was always the main metabolite. In conclusion OE2 regulation of uterine PG synthesis is depending on the animal species and cannot be explained by a unique effect on the cyclooxygenase, but rather by an interplay on the various enzymes of the arachidonic acid cascade.

Serum bile acids in newborns: evidence for an hepatic dysfunction in low-birth-weight infants.

The post-prandial pattern of total serum bile acids was studied in 47 newborns: 12 prematures (less than 36 weeks), 17 term low-birth-weight infants (less than the 3rd percentile), 18 term normals. The study was made at the end of the first month. Blood was collected in a peripheral vein using a microcatheter. Samples were taken at fasting time and 30, 60, 120, 180 min after a test meal intake (40 ml/kg of "humanized" milk based formula). Bile acids were assayed using an original enzymatic micromethod which needed only 50 microliter of serum and showed a sensitivity of 0.3 pmol in 200 microliter of reaction medium. The response of serum bile acids after the test meal was very similar in normal term newborns and in adults. Prematures exhibited bile acid levels slightly higher than normals, but this difference was significant only at 0 and 180 min. Low-birth-weight infants showed very high values of serum bile acids at all times during the test, compared to normal and premature infants. Serum levels of total bilirubin and alkaline phosphatase were similar in all 3 groups. These results are not consistent with cholestasis but rather indicate a specific dysfunction in bile acid metabolism in low-birth-weight infants.

[Mononucleated phagocytes from rheumatoid synovial fluid. Synthesis of prostanoids and first pharmacological applications]. / Phagocytes mononucléés du liquide synovial rhumatoïde. Synthèse de prostanoïdes et premières applications pharmacologiques.

An original and easily reproducible technique for isolating and culturating mononucleated phagocytes from rheumatoid synovial fluid was used by the authors to study the synthesis of PGE2, PGF alpha 2 and TxB2 by macrophages taken from 16 patients with rheumatoid arthritis. The functional importance of these macrophages was evaluated by comparing their ratios of prostaglandin synthesis. PGE2 and TxB2 are released in large quantities and are increased after stimulation with zymosan. Two distinct cell populations having different biochemical characteristics were identified from these macrophages based on their capacity to synthesize prostaglandins. Prostaglandin synthesis was correlated with the clinical progression of rheumatoid arthritic disease as evaluated by Lee's index and the time required to loosen up morning joint stiffness in these patients. No correlation was found between prostaglandin synthesis and the quantity of synovial fluid in the affected joint. The first results of a pharmacological study evaluating the activity of non steroid anti-inflammatory drugs using this experimental model are presented here. The addition of indomethacin to culture medium produces dose-dependent inhibition of prostaglandin synthesis.

Mononuclear phagocytes, prostanoids and rheumatoid arthritis.

We describe a new, simple procedure for obtaining and studying mononuclear phagocytes from inflammatory synovial fluid. The viability and functional characteristics of these macrophages have been carefully checked. This technique has been used to evaluate the macrophagic synthesis of prostaglandins in 16 patients with evolutive rheumatoid arthritis. Significant amounts of prostaglandins, mainly PGE2 and TxB2, are released and increased after zymosan administration. The three tested prostaglandins (PGE2, PGF2 alpha and TxB2) responded in a similar manner. The in vitro PG biosynthesis and the number of harvested adherent cells are negatively correlated with the synovial fluid volume but positively correlated with the disease activity. In fact, we found an heterogeneity among the analyzed macrophages; these can be divided into two populations with marked differences in their PG biosynthesis. The one with the highest synthesis corresponded to the more severe disease.

Plasma lipoperoxides and aging. Critical assessment of the thiobarbituric acid method for the measurement of lipoperoxides and malondialdehyde.

Thiobarbituric acid reactive products were measured in plasma of 66 normal subjects ranging from 19 to 92 years of age. A significant correlation (r = 0.54, p less than 10(-6) was formed between the level of these metabolites and the age of the subjects. As a matter of fact this method analyzes two kinds of substances from two different origins, free malondialdehyde or lipid peroxides. These two fractions were identified by extracting lipids using Folch's method and leaving all free malondialdehyde in the plasma (95.6 +/- 11.8% recovery for a known amount added to the plasma). Plasma analysis prior to and after extraction showed that thiobarbituric acid reactive products were lipid peroxides for 74.8 +/- 4.3%, the remainder being free malondialdehyde. The origin of these two fractions is discussed.

[Alveolar macrophages in asthmatic patients: study of their phagocytic function]. / Macrophages alvéolaires de l'asthmatique: étude de leur fonction phagocytaire.

Alveolar macrophages (AM) form the first line of defense against aggression in the lungs. The goal of the present study was to evaluate AM phagocytic function in two types of asthmatic patients: allergic asthma (14 patients) and asthma following ingestion of aspirin (6 patients). Macrophages obtained from bronchiolo-alveolar washings were isolated as a result of their adherence to glass and cultured for 24 hours. The results show that AM viability and phagocytosis of zymosan are reduced in the asthmatic patient. Decreased viability in patients sensitive to aspirin may be the result of alveolar eosinophilia, a quasiconstant finding in asthmatics. Although impaired phagocytosis of zymosan may be explained by the presence of surface IgE on AM of the allergic asthma patient, this is not the case for the aspirin sensitive patient.

[Technic for isolation and culture of macrophages from human synovial fluid]. / Technique d'isolement et de culture des macrophages du liquide synovial humain.

Macrophages from synovial fluid were obtained by joint puncture of patients with rheumatoid arthritis or non-rheumatoid exudative arthropathies. This fluid was subsequently centrifuged and the resulting cell pellet was recovered and distributed for in vitro culture (Falcon Petri dishes: 60 x 15 or 35 x 10). Cells were washed after 4 hours in culture to eliminate non-macrophagic cells. A second cell wash was effected 24 hours later to eliminate non-adherent cells. The culture medium employed was fetal calf serum MMP 119 (Gibco). The macrophagic nature of the cultured cells was ascertained by non specific esterase determinations (Burstone method) and estimation of the phagocytic index (using zymosan). This technique permits the conservation of viable monocytes in culture thus facilitating investigation of their metabolic activity (for example: prostaglandins, prostanoids or fatty acids) and the effects of anti-inflammatory agents.