Poor performance of albumin or protein-adjusted plasma calcium to diagnose dyscalcemia in hospitalized patients: A confirmatory study in a general internal medicine department.

INTRODUCTION: Hypo- and hypercalcemia are common and some causes require urgent diagnosis and treatment. Measurement of ionized calcium is the reference test to diagnose calcium disorders but total calcium adjusted for protein or albumin concentration is more often used. METHODS: Patients hospitalised in a general internal medicine department from September 2013 to December 2015 who had a total plasma calcium concentration and a serum albumin or protein concentration measured within 24h of a ionized calcium blood measurement were included. Total calcium was adjusted for protein or albumin concentration using widely used formulas and compared to ionized calcium as the gold standard. RESULTS: Among 210 included patients, 46 (22%) had hypocalcemia, 124 (59%) normocalcemia and 40 (19%) hypercalcemia according to ionized calcium concentration. Total calcium had 50% sensitivity and 95% specificity to diagnose hypocalcemia and a 93% sensitivity and 89% specificity to diagnose hypercalcemia. Adjusting total calcium for protein or albumin concentrations did not increase and sometimes decreased diagnostic accuracy. CONCLUSION: Total calcium, with or without albumin/protein adjustment, is poorly sensitive to screen for hypocalcemia. Unadjusted total calcium is as sensitive as protein- or albumin-adjusted total calcium to screen for hypercalcemia. These data argue against the use of albumin- or protein-adjusted calcium. Ionized calcium measurement should be performed to confirm dyscalcemia in patients with abnormal total calcium concentration and to rule out hypocalcemia in patients with total calcium concentration in the lower range of normal values.

Effect of ZnO Nanoparticle Content on the Structural and Ionic Transport Parameters of Polyvinyl Alcohol Based Proton-Conducting Polymer Electrolyte Membranes.

Proton conducting nanocomposite solid polymer electrolytes (NSPEs) based on polyvinyl alcohol/ammonium nitrate (PVA/NH4NO3) and different contents of zinc oxide nanoparticles (ZnO-NPs) have been prepared using the casting solution method. The XRD analysis revealed that the sample with 2 wt.% ZnO-NPs has a high amorphous content. The ionic conductivity analysis for the prepared membranes has been carried out over a wide range of frequencies at varying temperatures. Impedance analysis shows that sample with 2 wt.% ZnO-NPs has a smaller bulk resistance compared to that of undoped polymer electrolyte. A small amount of ZnO-NPs was found to enhance the proton-conduction significantly; the highest obtainable room-temperature ionic conductivity was 4.71 × 10-4 S/cm. The effect of ZnO-NP content on the transport parameters of the prepared proton-conducting NSPEs was investigated using the Rice-Roth model; the results reveal that the increase in ionic conductivity is due to an increment in the number of proton ions and their mobility.

On-Site Fecal Sludge Treatment with the Anaerobic Digestion Pasteurization Latrine.

The Anaerobic Digestion Pasteurization Latrine (ADPL) is a self-contained and energy neutral on-site sanitation system using anaerobic digestion of fecal sludge to generate biogas and then uses the biogas to pasteurize the digester effluent at 65-75°C to produce a safe effluent that can be reused locally as a fertilizer. Two ADPL systems were installed on residential plots with 17 and 35 residents in a peri-urban area outside of Eldoret, Kenya. Each system comprised three toilets built above a floating dome digester and one heat pasteurization system to sanitize the digested effluent. ADPLs are simple systems, with no moving parts and relying on gravity-induced flows. Adoption at the two sites was successful, and residents reported that the systems had little to no odor or flies. ADPLs were monitored for biogas production and temperatures in the pasteurization system. ADPLs serving 17 and 35 residents produced on average 16 and 11 Lbiogas/person/day (maximum of 20 and 15 Lbiogas/p/d), respectively. The temperature in the sterilization system was greater than 65°C on 58% and 87% of sampling days during the most stable period of operation. Treated effluent was analyzed periodically for chemical oxygen demand (COD), biochemical oxygen demand (BOD), total ammonia nitrogen (TAN), pH, and fecal coliform (FC). On average, the effluent at the two locations contained 4,540 and 6,450 mg COD/L (an 85% or 89% reduction of the estimated input), 2,050 and 3,970 mg BOD/L, and 2,420 and 4,760 mg NH3-N, respectively, and greater than 5 log reductions of FC (nondetectable) in the sterilization tank. Results from this field study show that anaerobic digestion of minimally diluted fecal sludge can provide enough energy to pasteurize digester effluent and that the ADPL may be a suitable option for on-site fecal sludge treatment.

Skin necrosis due to fluindione treatment: a rare but serious complication.

In the setting of protein C deficiency, skin necrosis, which occurs most often at the initial phase of oral anticoagulants therapy, is a rare side effect. Six cases have previously been reported in the literature. In this case report, we present a protein C deficient 42-year-old woman who was being treated for venous thrombosis. Five days after the initiation of oral anticoagulant treatment, she developed extensive skin necrosis on her left calf, followed by a painful leg ulcer. The pathogenesis underlying skin necrosis caused by anticoagulation therapy is still not clear. Despite only a few cases being reported in the literature, it is important to recognise this complication since adequate therapeutic approaches leading to a stable anticoagulation state may prevent it.

[Skin necrosis during long-term fluindione treatment revealing protein C deficiency]. / Nécroses cutanées tardives sous fluindione révélant un déficit en protéine C.

BACKGROUND: Cutaneous necrosis is a rare complication of vitamin K antagonist therapy. It presents as cutaneous hemorrhagic necrosis and usually occurs at the start of treatment. We describe an atypical case of recurrent skin necrosis after two years of treatment with fluindione. CASE REPORT: A 70-year old woman with a history of venous thromboembolism and obesity presented with a large haemorrhagic necrosis of the abdominal wall. She had been treated with fluindione for two years. Genetic protein C deficiency was discovered. Resumption of vitamin K antagonist therapy was followed by recurrence of skin necrosis despite concomitant administration of heparin. Treatment with vitamin K antagonists could not be continued. DISCUSSION: This observation is unusual due to the late onset of skin necrosis. The condition usually begins shortly after initiation of vitamin K antagonist therapy, generally between the third and the sixth day of treatment. It is due to a transient hypercoagulable state in patients with protein C deficiency or, in rare cases, protein S deficiency. This late-onset skin necrosis, occurring many years after initiation of anticoagulant therapy, may be explained by a sudden worsening of pre-existing protein C deficiency due to infectious and iatrogenic factors.

Protein S inherited qualitative deficiency: novel mutations and phenotypic influence.

BACKGROUND: Only a few mutations associated with qualitative protein S deficiency have already been described. Sensitivity and specificity for type II PROS1 mutations of commercially available reagents for measuring Protein S (PS) activity are not well established. Whether these mutations are significant risk factors for thrombosis remains an unresolved question. METHODS: In order to address the first point, we present and discuss the results of PROS1 analysis performed in the 30 probands with type II PS-inherited deficiency suspicion and 35 relatives, studied in our laboratory between 2000 and 2008. In order to investigate the influence of type II mutations on the coagulability level, thrombin generation tests were performed on plasma from 102 PROS1 type II, type I/III or PS Herleen mutation heterozygous carriers and controls. RESULTS: Mutations (12 novel, six already described) which probably explain the qualitative phenotype, were found in 27 (90%) out of the 30 probands studied. In relatives, 78% of heterozygotes presented with a type II phenotype. An APC resistance phenotype was documented in type II and type I/III defects heterozygous carriers; however, the effect of type II was milder than the effect of type I/III PS mutations. CONCLUSIONS: A PS functional assay (Staclot PS, Stago) was efficient in screening for PROS1 type II defects, particularly in probands. A significant positive influence of type II mutations on ex vivo thrombin generation was demonstrated. However, whether these mutations increase the risk of venous thromboembolism requires further investigation.

Gross deletions/duplications in PROS1 are relatively common in point mutation-negative hereditary protein S deficiency.

Hereditary protein S (PS) deficiency is an autosomal disorder caused by mutations in the PS gene (PROS1). Conventional PCR-based mutation detection identifies PROS1 point mutations in approximately 50% of the cases. To verify if gross copy number variations (CNVs) are often present in point mutation-negative hereditary PS deficiency we used multiplex ligation-dependent probe amplification (MLPA) as a detection tool in samples from individuals with a high probability of having true PS deficiency. To this end, DNA samples from nine PS deficient probands with family members (seven type I and two type III) and nine isolated probands (three type I and six type III), in whom PROS1 mutations were not found by DNA sequencing, were evaluated. An independent quantitative PCR (qPCR) was performed to confirm the findings of the MLPA assay. Family members were also tested when DNA was available. Gross abnormalities of PROS1 were found in six out of eighteen probands. In three probands complete deletion of the gene was detected. Two probands had a partial deletion involving different parts of the gene (one from exon 4 through 9 and another from exon 9 through 11). One family showed a duplication of part of PROS1. qPCR analysis was in accordance with these results. In conclusion, this study substantiates that gross gene abnormalities in PROS1 are relatively common in hereditary PS deficient patients and that MLPA is a useful tool for direct screening of CNVs in PROS1 point mutation-negative individuals.

[Multiple osteonecroses and venous thrombosis: one case of patient with a novel mutation of protein C (N102S) and heterozygous for FV Leiden]. / Ostéonécroses et thromboses veineuses multiples : un cas de patient porteur d'une nouvelle mutation de la protéine C (N102S) et hétérozygote pour le facteur V Leiden.

The association of a thrombo-embolic venous disease and multiple osteonecroses occurring in the presence of biological risk factors for thrombosis is rarely described in the literature. We report here the case of a 35-year old patient with such clinical manifestations. This patient is heterozygous for a novel mutation of the protein C gene (N102S) and for FV Leiden polymorphism. The clinical history is characterized by numerous thrombo-embolic venous episodes associated with several episodes of epiphysis osteonecrosis requiring two hip total prostheses and two knee total prostheses. The particular clinical features here are the multiple osteonecroses and the unusual localisation of brain and genital thromboses. The absence of both venous thromboembolic and osteonecrosis events in the relatives presenting the same genetic pattern suggests broad phenotype variations in the clinical expression of these genetic abnormalities. In osteonecrosis associated with thrombophilia, some authors have proposed treatment with stanazolol, which increase circulating protein C concentration. The effectiveness of this drug among such patients should be evaluated by clinical studies.

Diagnostic score for heparin-induced thrombocytopenia after cardiopulmonary bypass.

Heparin-induced thrombocytopenia (HIT) occurs in nearly 3% of patients treated with heparin after cardiopulmonary bypass (CPB). HIT carries a risk of severe thrombotic complications, and must be diagnosed rapidly. To identify simple criteria for estimating the probability of HIT after CPB, we retrospectively analyzed the files of 84 patients with suspected HIT after CPB and we analyzed the usefulness of several variables collected at the time of HIT suspicion to estimate HIT probability. HIT was confirmed in 35 cases and ruled out in 49 cases, on the basis of a platelet increment after heparin withdrawal, detection of heparin-dependent antibodies, and absence of other clear cause of thrombocytopenia. A biphasic platelet count from CPB to the first day of suspected HIT, an interval of >/= 5 days from CPB to the first day of suspected HIT, and a CPB duration of

Impact of progestagens on activated protein C (APC) resistance among users of oral contraceptives.

Oral contraceptive (OC) use is associated with an increased risk of venous thromboembolism. Previous data reported higher thrombotic risk in women using third-generation combined OC than in those using second generation OC. The difference could be explained by differential effects of progestagens on plasma sensitivity to activated protein C (APC). The main purpose of this cross-sectional study was to assess the influence of a progestagen-only OC (chlormadinone acetate) as well as the effect of several combined OC with different progestagen components on APC resistance. The effect of APC on endogenous thrombin potential (ETP) was investigated in the plasma of healthy women using either combined OC (n=82) or progestagen-only OC (n=28), and in non-users (n=64). Carriers of factor V Leiden were excluded. Compared with non-users, there was no significant change in APC resistance in women using progestagen-only OC. Women who used combined OC were less sensitive to APC than non-users (P < 0.001) and the difference was significantly more pronounced in women using third-generation OC (n=41) than in those who used second-generation OC containing levonorgestrel (n=22) (P < 0.05). Compared with OC containing levonorgestrel, use of norethisterone-containing OC (n = 9) was associated with an increased resistance to APC (P < 0.05). Women who used cyproterone-containing OC (n = 10) were less sensitive to APC than those using third-generation OC (P < 0.05) or second-generation OC containing levonorgestrel (P < 0.05). Protein S, factor II and FVIII levels explained in part the OC-related changes in APC sensitivity variations. ETP-based APC resistance may contribute to explain why different brands of OC can be associated with different levels of thrombogenicity.

The factor II G20210A gene polymorphism, but not factor V Arg506Gln, is associated with peripheral arterial disease: results of a case-control study.

BACKGROUND: The FIIG20210A polymorphism has been associated with arterial wall thickness and atherothrombotic diseases in selected subgroups. The FVArg506Gln polymorphism does not seem to be associated with arterial diseases. Few data are available on these polymorphisms and the risk of peripheral arterial disease (PAD). OBJECTIVES: To study the association between the FIIG20210A and FVArg506Gln polymorphisms and PAD and its clinical severity. To examine the potential interactions with traditional vascular risk factors. PATIENTS AND METHODS: We studied 184 consecutive male patients under 70 years of age with symptomatic PAD and 330 age-matched male controls free of symptomatic PAD and with no cardiovascular history. We evaluated the FIIG20210A and FVArg506Gln polymorphisms in all subjects. RESULTS: Mean age was 57.1 +/- 7.2 years (cases) and 56.7 +/- 7.6 years (controls). The FII20210A allele was more frequent in PAD patients with odds ratios (OR) of 3.77 (1.39-10.2) in univariate analysis and 4.30 (1.3-14.7) after adjustment for diabetes, smoking, hypertension and hypercholesterolemia. In smokers or past smokers the magnitude of the association was markedly increased but there was no evidence of an interaction between tobacco exposure and FIIG20210A. In case subjects, the FII20210A allele was also associated with critical ischemia [OR = 4.1 (1.1-15.7), P = 0.039 in multivariate analysis]. FVArg506Gln was not associated with PAD [OR = 0.65 (0.27-1.54) and 0.77 (0.28-2.1) in univariate and multivariate analyses, respectively]. CONCLUSIONS: The FIIG20210A gene polymorphism may be a risk factor for PAD and its severity. In contrast, the FVArg506Gln polymorphism is not associated with PAD.

[Genetics of venous thromboembolism]. / Génétique de la maladie thrombo-embolique veineuse.

Venous thrombo-embolic disease is a multifactorial disease which arises in situations of thrombosis risk with poorly understood genetic and/or acquired risk factors. Constitutional anomalies concerning five coagulation proteins (antithrombin, protein C, protein S, factor V and prothrombin) are established risk factors for which the harmful role has been demonstrated in thrombophilic families and/or in case-control studies. New studies (twin studies, GAIT study) which use joint subjects and appropriate statistical methods are aimed at a better understanding of the genetic component and the environmental component of thrombotic risk. They have allowed demonstration of the importance of the contribution of heredity to the variability of the coagulation parameter concentration and the variability of susceptibility to thrombosis, and have implied the influence of genes with pleiotrophic effect. The identification of these new loci will allow a better evaluation of the components of thrombotic risk and perhaps lead to individualised preventive therapeutic management.

Familial thrombophilia is an oligogenetic disease: involvement of the prothrombin G20210A, PROC and PROS gene mutations.

Population-based case-control studies and cases previously published suggest that the prothrombin G20210A mutation is a weak risk factor for thrombosis, leading to clinical expression mainly in the presence of other risk factors. We report the results of plasma and genetic analyses performed in a 13-year-old symptomatic boy homozygous for the 20210A allele and in his family, which are in accordance with this suggestion. These analyses demonstrated the presence of several PROC (R-5W, R87H) and PROS (R60C, T103N) gene mutations in this family. These additional mutations have modulating effects on clinical expression of the G20210A mutation. The present family study illustrates the concept of 'mild' mutation and the hypothesis that familial thrombophilia is a multifactorial disease.

[Analytical choices for biological tests in thrombotic pathology]. / Choix raisonné des examens biologiques dans la pathologie thrombotique.

The knowledge of genetic and occurred risk factors allows us to propose laboratory investigations in patients with thromboembolism (TE) to evaluate the risk for recurrence. Beside coagulation inhibitor deficiencies accounting for less than 10% of cases, factor V Leiden and prothrombin G20210A mutations are found in around 30% and 10% respectively. Increased factor VIII and hyperhomocysteinemia resulting from interaction of genetic factors and environmental factor are also risk factors for TE. Finally, antiphospholipid antibodies, a complex family of antibodies recognizing various phospholipid or phospholipid binding proteins, are also associated with TE.

Intracerebral hemorrhage associated with a novel antithrombin gene mutation in a neonate.

Fatal cerebral hemorrhage involving the left thalamus in a neonate was attributed to deep cerebral vein thrombosis. Although antithrombin levels were at the lower end of the normal range, family and genetic studies showed constitutional type I antithrombin deficiency related to a novel missense mutation in the antithrombin gene.