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The International osteoporosis foundation and the International federation of clinical chemistry (IFCC) Bone marker standards working group have identified N-terminal propeptide of type I procollagen (PINP) and C-terminal telopeptide of type I collagen (CTX-I) in blood to be the reference markers of bone turnover for the fracture risk prediction and monitoring of osteoporosis treatment. Although used in clinical research for many years, bone turnover markers (BTM) have not been widely adopted in clinical practice primarily due to their poor within-subject and between-lab reproducibility. The NBHA bone turnover marker project team aim to reduce pre-analytical variability of CTX-I and PINP measurements through standardized sample handling and patient preparation. Recommendations for sample handling and patient preparations were made based on review of available publications and pragmatic considerations to reduce pre-analytical variability. Controllable and un-controllable patient-related factors were reviewed to facilitate interpretation and sample collection. Samples for CTX-I must be collected consistently in the morning hours in the fasted state. EDTA plasma is preferred for CTX-I for its greater sample stability. Sample collection conditions for PINP are less critical as PINP has minimal circadian variability and is not affected by food intake. Sample stability limits should be observed. The uncontrollable aspects (age, sex, pregnancy, immobility, recent fracture, co-morbidities, anti-osteoporotic drugs, other medications) should be considered in BTM interpretation. Adopting standardized sample handling and patient preparation procedures will significantly reduce controllable pre-analytical variability. The successful adoption of such recommendations necessitates the close collaboration of various stakeholders at the global stage, including the laboratories, the medical community, the reagent manufacturers and the regulatory agencies.

Introduction: Can vitamin D deficiency be predicted by patient questionnaire? Does it lead to secondary hyperparathyroidism that may cause excessive bone resorption? We studied non-osteoporotic subjects in their fifties, in whom vitamin D levels are often tested., Patients and Methods: Patients hospitalised for degenerative osteoarthritis or consulting for assessment of menopause, without renal failure and not treated with vitamin D, completed a questionnaire on sun exposure and underwent measurement of serum calcium, creatinine, 25OH vitamin D, PTH and CTX., Results: Five hundred and twenty-six subjects, mean age 54.6 years (71% women), were investigated throughout the year. 25OH vitamin D levels were correlated with sun exposure and varied according to the month of the year, unlike PTH and CTX levels. From November to May, over 90% of subjects had 25OH vitamin D levels<30ng/mL. Of the subjects who did not expose their face, arms and legs to the sun for at least 20min/day, 94% had 25OH vitamin D levels<30ng/mL. PTH levels were negatively correlated with those of 25OH vitamin D. Serum CTX levels were not correlated with PTH or 25OH vitamin D. Only 13% of subjects presented with secondary hyperparathyroidism, characterised by serum calcium<2.55mmol/L and PTH>65pg/mL, associated with increased CTX levels., Conclusion: Vitamin D deficiency can be predicted by patient questionnaire. It very rarely leads to secondary hyperparathyroidism., (Copyright © 2015. Published by Elsevier Masson SAS.)

Blood testing using the dried blood spot (DBS) is used since the 1960s in clinical analysis, mainly within the framework of the neonatal screening (Guthrie test). Since then numerous analytes such as nucleic acids, small molecules or lipids, were successfully measured on the DBS. While this pre-analytical method represents an interesting alternative to classic blood sampling, its use in routine is still limited. We review here the different clinical applications of the blood sampling on DBS and estimate its future place, supported by the new methods of analysis as the LC-MS mass spectrometry.

Background: Currently, for the diagnosis of osteoporosis, we search risk factors and measure bone mineral density (BMD) by DXA method. However, bone turnover markers, unused still in practice, have shown an interest especially in the prediction of fracture risk. aim: To determine the relationship between bone markers, BMD and osteoporotic fracture. methods: Prospective study of 65 women referred for measure of BMD during the period between May and August 2010. Each patient had a dosage of serum bone formation markers: osteocalcin (OC) and N-terminal propeptide of type I collagen (P1NP) and bone resorption markers: serum and urinary C-terminal telopeptide of type I collagen (β-CTX or CrossLaps) as well as parathyroid hormone and calcium. Risk factors of osteoporosis were identified in each case. results: Our 65 women had a mean age of 58.6 ± 12.1 years. The majority (83%) were menopausal women. Osteoporosis was found in 52%, osteopenia 26% and normal BMD 22% of cases. An increase in bone turnover markers was correlated with menopause (p = 0. 001 for the OC, p = 0.016 for urinary CTX), a low body mass index (p = 0.015 for OC, p = 0.042 for serum CTX) and osteoporosis (p <0.001 for P1NP, p <0.001 for serum and urinary CTX). Corticosteroid therapy was correlated with a decrease in bone formation markers (p = 0.002 for P1NP). The presence of fracture was only associated with increased urinary CTX (p = 0.05)., Conclusion: Bone turnover markers increase in menopausal women and in case of low BMD. However, their contribution in the diagnosis of osteoporosis is low. They are rather an interest in the prediction of fracture risk.

Background: These twenty last years, the metabolic syndrome was accused in various human pathologies including android obesity and type 2 diabetes. In obesity, increased body weight is frequently associated with excessive caloric food and sedentary activities. The Behaviour Eating Disorders (BED) is involved in over-consumption alimentary. In Algeria, we observed increasingly deviations in life-style alimentary, from the Mediterranean-Cretan model to American fast-food model., Aim: To study interactions between the BED, hormonal secretions and metabolic syndrome parameters., Methods: The present study was undertaken on Algerian population cohorts composed of 30 obese subjects, 70 type 2 diabetic patients and 30 healthy subjects. The BED was evaluated by TFEQ and DEBQ tests. Anthropometric parameters (waist circumference, IMC), metabolic parameters (glucose, triglycerides, HDL-C, LDL-C) and hormones (insulin, peptide C, ACTH, cortisol, GH) were determinates by biometrics, spectrophotometry and radioimmunology methods, respectively., Results: Multivariate analyses showed the high correlation between the BED and the metabolic syndrome, particularly a critical insulinoresistance state (IR). This IR generates in periphery a whole of metabolic disorders: dyslipidemia, hyperglycemia and hypertrophy of adipose tissue. In diabetic and obese patients, cortisol, ACTH and GH secretions are insidiously altered and to lead metabolic disorders., Conclusion: In this study, the role of the BED in obesity and diabetes genesis seems to be confirmed. In response to nutritional stress, the BED generates a hyperactivity of endocrine pancreas, adrenal gland, and pituitary gland. It appeared that Algerian population is not adapted to fast-food American model.

Abdominal -- and not peripheral -- obesity induces insulin resistance. Morbid obesity is not always accompanied by either diabetes mellitus or metabolic syndrome. Development of morbid obesity can require appropriate insulin secretion and recruitment of small insulin-sensitive adipocytes, able to store fatty acids. These fatty acids are therefore not stored in ectopic sites (muscle, liver, islets of Langerhans), and neither insulin resistance nor glucolipid toxicity develops and causes insulin deficiency. This explains the relative rarity of diabetes in morbid obesity. Patients with morbid obesity are at greater risk of developing mechanical complications (e.g. cardiac, pulmonary, or locomotor system, or sleep apnea) than metabolic complications or cardiovascular heart disease.

Aims: To evaluate the effect of the physical activity on bone turnover in young male soccer players at the Tanner's stage of 1-2., Material and Methods: 61 young soccer players (13,4 +/- 0,3 years old) who actively participated in soccer since 3,7 +/- 0,7 years were compared to 60 age and sex- matched non active subjects. Bone mineral density (BMD) of whole body, and in specific skeleton sites, fatty body mass (FBM) and lean body mass (LBM) were determined by a dual energy X-ray absorptiometry (DXA). Total plasma alkaline phosphatase (ALP) and plasma bone alkaline phosphatase (BALP), plasma osteocalcin (OC) and plasma collagen type I cross-linked C-telopeptide (CTX) were measured., Results: BMD of the whole body and at the lumbar spine (L2-L4), femoral, lower limbs and LBM were significantly higher in young soccer players than in controls. The biochemical markers of bone turnover: ALP (6,7%), BALP (8,9%), OC (3%) and CTX (3,1%) were not significantly higher in sportsmen than in controls. The calcium was significantly higher in sportsmen than in controls., Conclusion: These results suggest that soccer practice induced an increase of bone mass in boys. The increase in the level of bone turnover evaluated by the new biochemical markers was not significant in the sportsmen.

The recent development of new biochemical markers has considerably improved the non invasive exploration of bone turnover. Currently, the most sensitive markers to access bone formation are osteocalcin, bone alkaline phosphatase and N-terminal propeptide of type I procollagen. Blood or urinary immunoassays of pyridinoline, deoxy-pyridinoline and terminal telopeptides type I of collagen are currently the best indices to evaluate the bone resorption. The principal application fields of biochemical markers in postmenopausal osteoporosis, in combination with the measurement of the bone mineral mass, are primarily the monitoring of anti-resorptive therapy response and prediction of bone loss and risk of fractures. In fact, treatment with anti-resorptive drugs is followed by rapid decrease of bone markers levels (3 months for the markers of resorption and 6 months for those of osteo-formation ), whereas the bone mineral density measurements requires at least two years to change significantly.

Heparin-induced thrombocytopenia remains a topical subject for at least two reasons. The first reason is the increasing prescription of low molecular weight heparins (LMWH) rather than unfractionated heparins, with limited laboratory surveillance, raising the question concerning the need for twice-weekly platelet counts, according to the recommendations of the Vidal drug directory. The second reason is the recent release onto the market of two products, danaparoid (Orgaran) and lepirudin (Refludin) for this precise indication of heparin-induced thrombocytopenia. These products greatly facilitate the management of this complication. Many basic research teams are trying to optimize the detection of heparin-dependent antibodies and to more clearly elucidate the mechanism of this particular thrombocytopenia, which carries a risk of very severe thrombotic complications when the diagnosis is delayed.

This study concerns 45 patients group one suffering from broncho-pulmonary cancer, the diagnosis was obtained by bronchial biopsies or by transparietal puncture using a scanner: there were 35 non-small cell bronchial carcinomas (CNPC) and 10 small cell bronchial cancers (CPC). The control patients (99 patients) were divided up as follows: 44 pleuro-pulmonary infections (group two) and 55 with respiratory failure of various causes other than infectious episodes (group three). In group one the level for TPA was positive in 30 cases (the threshold value was 90 units per litre), 9 for CA 19.9, 7 for ACE and 9 for NSE. The overall sensitivity was thus better for TPA. There was no correlation between TPA and type of tumour histology nor between the different markers. Their association did not improve the sensitivity. The NSE however, remained the most sensitive test for the diagnosis of CPC with six positive tests out of ten. In the control population, the specificity of TPA (66%) was less than that of ACE (100%) or of CA 19.9 (94%) and the false positives were significantly more numerous in group two: 21 patients had a positive test compared to only 12 in group three. Finally we noticed an increase in the level of TPA contrary to other markers, as a function of the extent of the disease from the carcinoma (CNPC unique). The TPA is thus the most sensitive and it turns out to be better reflector to the extent of the tumour disease than either ACE, CA 19.9 or NSE but this applies uniquely to non-small cell carcinoma.

During pathologies such as arteriosclerosis and emphysema, degradation of elastin by elastases occurs and elastin peptides are produced. In order to evaluate elastin degradation, measurements of elastin peptide concentration in human blood were carried out. According to elastin peptides used for obtention of antibodies and for ELISA, the measured values are different. Elastin peptides have several biological effects: they are chemotactic, modify ion fluxes and several intracellular mechanisms.

Parallel time courses of clinical and behavioural parameters and levels of plasma substance P-like immunoreactivity (SPLI-PI) were studied in arthritic rats (adjuvant induced arthritis, AIA, a chronic pain model). Acute (14 and 21 post-inoculation days,PI) and post-acute (42 days PI) phases of the syndrome were investigated. These data were compared with those obtained in a control situation (inoculation day). In a second experimental series, levels of substance P-like immunoreactivity in cerebrospinal fluid (SPLI-CSF) were determined at the same stages of AIA. In arthritic rats SPLI-PI was strongly enhanced (X4) as early as 14 days PI and remained increased (X4) at all stages studied, whereas SPLI-LCR was significantly increased (X2) only 21 days PI and returned to control levels at 42 days PI. These data suggest that SP could be distributed in two different pools, a peripheral one of inflammatory origin, and a central one which could be more specific to the chronic pain situation.

Fifty nine insulin dependant diabetics were hospitalised for a trial withdrawal of insulin: 17 patients rapidly showed signs of lack of insulin, 18 did not develop cetoacidosis but could not be stabilised on diet and oral hypoglycemic agents, 24 were stabilised without insulin. A statistical study (multifactorial analysis of correlations, plotting of ROC graphs) validated the classification of these diabetics into 3 groups. It also showed that in patients with hypoglycemia, the values of C-protein, and after intravenous injection of tolbutamide, were good predictive factors for insulin-dependance: all patients with basal C-protein less than 1,9 ng/ml could not be stabilised without insulin; when the basal C-protein greater than or equal to 1,9 ng/ml and the amplitude of response at the 5th min was greater than or equal to 0,4 ng/ml, the diabetes could be stabilised by diet and oral hypoglycemic agents in 90 p. 100 of cases. This institutes an easy, reliable and economic method of detecting abusive insulin therapy.

Platelets carry, and release after stimulation, mitotic activities for connective tissue cells in culture. One of these activities has been studied carefully: the PDGF, it is a peptide, molecular weight 30,000, pHi 9.8-10, heat stable. Beside this mitotic activity, PDGF bears migrative, chemotactic and metabolic functions. Hypothetical theories have implicated PDGF in different pathological conditions (atherosclerosis, myelofibrosis, ...) but there is no direct proof that PDGF is active in vivo. Recently there has been experimental data suggesting that the mitotic activity of PDGF in vitro could be a technical artefact.

A new method of intraperitoneal transplantation of the whole pancreas as donor-pancreaticoduodeno/recipient-cholecystostomy is described. The pancreas with duodenal segment is placed into the dorsal peritoneal cavity and the portal vein is anastomosed with the subhepatic vena cava and the long aortic segment of the graft with the distal aorta of the recipient. An anastomosis between the donor's duodenal segment and the recipient's gallbladder ensures exocrine pancreatic secretion drainage.

Insulin secretion is assessed by simultaneous radioimmunological assay of insulin (R.I.I.) and C-peptide (R.I.C.P.) levels under basal conditions, and after stimulation by oral or intravenous glucose administration. Subjects with abnormal glucose tolerance demonstrate an increase in the ratios R.I.C.P. divided by glucose and R.I.C.P. divided by R.I.I. after fasting, increase in R.I.I. and R.I.C.P. occurring later and lasting longer after glucose loading. These anomalies are observed in both obese subjects and those with normal body weights. Simultaneous determination of R.I.I. and R.I.C.P. levels appears of value in a limited number of cases where glucose loading tests do not supply precise information on the quality of glucose tolerance. Obese subjects, in whom abnormal hyperglycemia levels provoked by oral glucose are observed, but in whom the peripheral glucose assimilation coefficient is normal, can be considered to be non-diabetic as shown by the levels of R.I.I. and R.I.C.P. and more particularly by the molar ratio R.I.C.P. divided by R.I.I.

SDS-polyacrylamide gel electrophoresis of solubilized whole platelets and isolated platelet membranes from patients with chronic myeloid leukemia (CML) as compared to the normal platelet-Schiff-stained glycoproteins (GP) (noted GP I, II, III of app. MW 155, 130 and 100.10(3) daltons, respectively), showed the following modifications: the decrease of GP I; the presence of the GP II and GP IIx doublet; the moderate and variable decrease of the GP III; the increase of the platelet glycolipid. No significant differences were detectable on the polypeptide patterns from CML platelets in respect to the normal 20-25 protein constituents. Using a radio-isotopic method it has been measured as an elevated catalytic transfer of the galactosyl and N-acetylgalactosaminyl residues from nucléotide 14C sugar derivatives onto the endogenous acceptors of platelets from CML. The results obtained suggested that CML platelets with recognized abnormal functions, have modifications at the level of the macromolecular surface glycoprotein constituents and/or changes of the glycoconjugates reactive structures involved in the devopment of the periodic acid-Schiff reaction as also in the transfer reaction of the sugar residues.

The peptide C, polypeptide secreted by the pancreas at the same time as insulin, presents a great interest in the evaluation of diabetic patients. First it allows a differentiation between insulin dependent diabetes (IDD) and non insulin dependent (NDD). A low and non stimulated levels of peptide C signifies an insulin dependence. Within the group of IDD patients the peptide C was low when the diabetes was discovered at a younger age and its secretion diminished as the diabetes progresses. The peptide C has also a prognostic interest in IDD. Low and non stimulable levels of peptide C signifies a difficult control of diabetes which needs two injections per day while high and stimulable levels will be seen in diabetes easy to control with one injection of insulin. Finally, values of peptide C does not permit to predict the onset of diabetic complications (retinopathy, acidocetosis) as well as the control of patients.

Combined continuous subcutaneous insulin infusion (CSII) and metformin (M) was tested in 10 overweighted insulin requiring diabetic patients (body mass index 27.9 +/- 4.9 kg/m2). They were still poorly controlled (HbA1 9.6 +/- 0.8%) despite large doses of lente insulin (Novo) (51.7 +/- 19.6 IU/24 h) injected once daily. With CSII after two weeks on placebo (P) they were enrolled in a randomized double blind cross-over trial with two successive one month periods of M (2550 mg/day) or P. At the end of the two-weeks period CSII--initial P, the daily regular insulin requirement decreased significantly (40.1 +/- 18.1). During M and P body weight and HbA1 were unchanged (respectively 28.6 +/- 6.0 vs 29.0 +/- 59 kg/m2 and 7.7 +/- 1.1 vs 7.7 +/- 0.8%). With M daily insulin requirements decreased significantly (32.0 +/- 16.8 vs 38.4 +/- 18.2 IU, p less than 0.05). During test-meals, M compared to placebo also reduced peripheral free insulin concentrations (-24.9 +/- 26.0%) while plasma glucose and C peptide remained comparable. In the conditions of this study, combined CSII and M reduced the insulin resistance observed in overweighted insulin requiring diabetic patients.

Plasma levels of insulin, C-peptide and glucose were monitored for 24 hours after a single injection of a mixture of regular and protamine zinc insulin in 22 diabetic children. Results were compared with those in 7 control children. They indicate that plasma insulin in diabetic children is similar or higher than that in controls over the entire time of survey. Plasma insulin is usually high in the first part of the day. However, important variations were observed from one patient to another and, in the same patient, from one day to another. These results stress the necessity of a daily individual adaptation of the doses in diabetic children and adolescents treated with one or several daily injections of insulin.

Basal and reactive peripheral hyperinsulinism recorded in alcoholic hepatic disease may result from decreased hepatic breakdown or pancreatic hypersecretion. C-peptide (CPR) and insulin (IRI) concentrations were measured in 3 groups of 8 alcoholic patients--steatosis, compensated and decompensated cirrhosis--and compared with 8 normal subjects in order to determine the importance of these two possibilities. At basal state, the molar ratio CPR/IRI was near the normal (8.7 +/- 0.9) but is diminished in the 8 hyperinsulinaemic patients (5.9 +/- 0.6). After i.v. glucose tolerance test and tolbutamide stimulations, an hyperreactivity of IRI and CPR may be noted in cirrhotics. A relative insensitivity of the B-cell to glucose appeared after comparison with the effect of tolbutamide. Thus basal hyperinsulinism resulted of decreased hepatic breakdown and stimulated hyperinsulinism resulted of hypersecretion. Glucose intolerance and anomalies of the insulin secretion were more apparent with severe hepatic disease.

The C-peptide is a polypeptide generated from enzymatic cleavage of proinsulin into insulin in pancreatic B. cell. C-peptide and insulin are secreted in an equimolar ratio. For this reason. C-peptide radio-immunoassay in blood or urine reflects the insulin secretion when direct insulin determination cannot be done (insulin treatment, circulating insulin antibodies). This assay is mainly used in clinical and investigational studies of insulin dependent diabets. It enabled demonstation of residual secretion of insulin in numerous insulin dependent patients (70% during the first year and 15% after the 15th year of the disease). Persistence of insulin secretion may play a role in the natural history of diabetes since patients with detectable C-peptide immunoreactivity have more stable diabetes, are controlled by lower insulin dose and are less ketosis prone than the others. Factors which influence persistence of B. cell activity in some IDD remain unknown.