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Serum levels of alpha-1-antitrypsin were measured by radial immunodiffusion, and phenotypes were determined by electrofocusing in acrylamide gel in 160 subjects who were used as controls in a case-control study of hepatocellular carcinoma (HCC). The results were studied in relation to age, sex, diagnostic category, tobacco smoking, consumption of alcoholic beverages, presence of hepatitis B surface antigen (HBsAg), and serum levels of alphafetoprotein (AFP) by modeling the data through multiple regression. There was no relation of serum alpha-1-antitrypsin values with sex, HBsAg, AFP, consumption of alcoholic beverages, and diagnostic category (p |is greater than~ 0.25). By contrast, there were statistically significant dose-dependent positive associations of serum alpha-1-antitrypsin with age and tobacco smoking (p |is less than~ 0.01 in both instances). The positive association of serum alpha-1-antitrypsin with tobacco smoking and the previously reported excessive elevation of serum alpha-1-antitrypsin in hepatitis B-negative tobacco-related cases of HCC suggest that alpha-1-antitrypsin is intimately related to the pathogenetic process linking tobacco smoking to HCC., Alpha-1-antitrypsin is one of at least nine plasma proteins that have been characterized as protease inhibitors (Harpel 1983). Alpha-1-antitrypsin is under genetic control, and more than 50 codominant alleles at [...]

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The IgG pattern in CSF was studied in 11 patients with multiple sclerosis who exhibited an oligoclonal banding upon thin-layer polyacrylamide gel isoelectric focusing followed by silver stain of unconcentrated CSF. Each patient received beta-interferon intrathecally during a 2 month period. No modification was observed over a 6 month period. In addition, the oligoclonal pattern was remarkably unique for each individual representing a typical "fingerprint" which allowed the identification of any single CSF. [ABSTRACT FROM AUTHOR]

We report 12 cases of lymphomas which occurred among 1670 patients with kidney or combined renal and pancreatic transplantation. Group 1 comprised nine patients presenting with the diffuse form of the disease where immunoblasts or mature plasma cells massively infiltrated all organs. The first symptom was a viral syndrome, associated with a restriction of heterogeneity of immunoglobulins; oligoclonal to monoclonal peaks of immunoglobulins appeared about 50 days after transplantation. All patients received antilymphocyte globulins (ALG), and seven were treated with cyclosporin. EBV infection could be demonstrated in almost all patients; three EBV lymphoblastoïd cell lines were established, their HLA phenotype being the same as the recipient of the graft. All patients finally died with renal and hepatic failure. Group 2 comprises three patients who presented solid B cell tumours of tonsils, lungs, and spleen at onset, extending to liver, kidney graft, lymph nodes, and brain. All received cyclosporin; two patients were treated with ALG, and one with OKT3. Immunoglobulins were polyclonal, oligoclonal, or decreased. Cell surface immunoglobulins were monoclonal on two tumours. EBV-DNA was positive within two tumours. Two patients presented EBV and CMV primary infection. CD4+T lymphocytes subsets were diminished at onset, and increased after cessation of immunosuppressive therapy. One patient died because of brain involvement; the two others are alive, one with perfect graft function. Therapy consisted of stopping immunosuppressive treatment, Acyclovir, and in two patients of group 2, monoclonal antibodies to pan-B and EBV receptor antigens. [ABSTRACT FROM PUBLISHER]

The two major protease inhibitors in human serum, alpha1-antitrypsin (α1AT) and alpha2- macroglobulin (α 2m), were analysed for their effects on antibody-dependent cell-mediated cytotoxocity (ADCC) and natural killing (NK). The results indicate that both α 1AT and α 2m decrease ADCC and NK in a dose-responsive pattern. [ABSTRACT FROM AUTHOR]

Introduction La thrombopénie auto-immune idiopathique (TAI) est une maladie d’évolution imprévisible, avec un passage à la chronicité dans plus de 80 % des cas. Il n’existe à ce jour aucun biomarqueur prédictif d’une rechute. Nous avons étudié les chaînes légères libres (CLL) sériques Kappa et Lambda comme facteur pronostique dans la TAI. Patients et méthodes Dans une étude rétrospective transversale, nous avons testé les sérums de patients recueillis en sérothèque entre 2006 et 2013 et conservés à −20 °C. Les patients inclus dans l’étude avaient plus de 18 ans et une TAI idiopathique. Ont été exclus les patients insuffisants rénaux et les thrombopénies secondaires (hémopathies malignes, lupus, VIH, VHC ou VHB). Les CLL kappa (K), et lambda (L), ont été dosées par le test FREELITE™ (The Binding Site, Birmingham, Royaume-Uni). La somme (K + L) ainsi que le rapport K/L ont été calculés puis comparés aux valeurs obtenues dans une population européenne saine de 282 individus. La rechute de la TAI était définie par une concentration plaquettaire < 30 G/L. Le seuil pronostique optimal des CLL était déterminé par courbe ROC. Les comparaisons de moyenne ont été faites par les tests de Student ou de Mann-Whitney, les comparaisons de proportion avec le test exact de Fisher. Les analyses de survie ont été réalisées selon les modèles de Kaplan-Meier et de Cox. A été considéré comme statistiquement significative une valeur de p < 0,05. Résultats Parmi les 66 patients (âge 45,2 ± 19,9, 41 femmes) inclus, 55 patients avaient un dosage plaquettaire concomitant du dosage des CLL (moyenne 34 G/L ± 23 ; min 5–max 99 G/L). Les plaquettes étaient entre 30–100 G/L chez 26 (47 %) patients. Aucun n’avait un taux de plaquettes > 100 G/L. Quarante et un dosages de CLL (62 %) étaient réalisés au diagnostic, 15 patients avaient été traités précédemment (corticoïdes n = 15, splénectomie n = 5, rituximab = 3, azathioprine = 2). Cinquante-deux (83 %) patients ont été suivis (médiane 2,8 ans, min 0,1–max 13,1) parmi lesquels 40 (76 %)ont reçu un traitement (en monothérapie : corticoïdes n = 9, immunoglobulines n = 1, dapsone n = 1 ; en association avec les corticoïdes, immunoglobulines, n = 24, rituximab n = 4, ou splénectomie n = 1). La durée médiane de la maladie avant la réalisation du dosage des CLL était de 0 année (min 0,0, max 60,1). Par rapport aux témoins, les patients avaient une augmentation significative ( p < 0,0001) des concentrations en CLL K (15,1 ± 7,35 versus 8,4 ± 4,5), et L (15,9 ± 5,7 versus 13,4 ± 5,2) et une augmentation significative de la somme (K + L) (31,1 ± 12,0 versus 21,8 ± 9,0), et du ratio K/L (0,95 ± 0,25 versus 0,63 ±0,23). Un ratio K/L > 1 a été retrouvé chez 42 % des patients et 6 % des témoins ( p < 0,0001). Aucune corrélation n’a été retrouvée entre la concentration des gammaglobulines (moyenne 15,5 ± 0,8 g/L) et celle des CLL K ( p = 0,41) et L ( p = 0,64). De même, aucune corrélation n’a été observée entre les concentrations des CLL K ( p = 0,88), L ( p = 0,59), la somme K + L ( p = 0,71), et le ratio K/L ( p = 0,42) et la concentration des plaquettes lors du dosage initial. Un ratio K/L > 1 était associé à un sur-risque de rechute de la TAI (18 % versus 50 % de rechutes, délai de rechute moyen médian 85 versus 39 mois ; HR 4,1, IC 1,48–11,4, p = 0,02) pour l’ensemble des pa [ABSTRACT FROM AUTHOR]

Introduction L’élévation des immunoglobulines (Ig) G4, sous classe d’IgG, est associée à différentes situations pathologiques. La maladie associée aux IgG4 est caractérisée une infiltration tissulaire lymphoplasmocytaire évoluant le plus souvent vers la fibrose et une élévation des IgG4 sériques > 1350 mg/dL. Ce taux n’est pas spécifique de la maladie liée aux IgG4, et Ebbo et al. ont décrit une augmentation dans de nombreuses autres situations notamment la M (3,4 % dans cette étude) [1] . En 2014, Mavragani et al. Note un taux élevé d’IgG4 chez 7,5 % des patients présentant un syndrome de Gougerot-Sjögren primaire, et identifiait ce sous-groupe comme à risque d’atteintes viscérales biliaires, pancréatiques et rénales [2] . Dans la M, plusieurs études ont retrouvé une élévation des IgG4 sériques chez les patients infectés ou colonisés à Pseudomonas aeruginosa avec défaut d’opsonisation bactérienne et lors des manifestations d’hypersensibilité immédiate [3] . Ces études soulevaient la question du rôle de l’infection chronique sur cette élévation des IgG4. L’objectif de notre travail est d’évaluer la prévalence d’une augmentation des IgG4 dans la M, et sa relation avec la situation clinique, microbiologique et pronostique. Patients et méthodes Il s’agit d’une étude de cohorte prospective, monocentrique, et transversale, incluant les patients de plus de 18 ans suivis pour une M dans le centre ressource et de compétence (CRCM) du CHU de Lyon au cours de l’année 2013. Les patients greffés ont été exclus. Les données cliniques, biologiques et microbiologiques ont été rétrospectivement recueillies, en particulier le genre, l’âge, l’IMC, l’atteinte des fonctions respiratoires, pancréatiques et endocriniennes ainsi que les manifestations d’hypersensibilité. Les patients ont été analysés en deux groupes, ceux présentant un taux IgG4 (immunoturbidimétrie) supérieur à 1350 mg/dL (IgG4+), et les patients présentant un taux IgG4 inférieur à 1350 mg/dL (IgG4−). Les comparaisons ont été réalisées avec le test de Fisher ou du Chi 2 pour les proportions, et par le test de t -Student ou de Mann-Whitney pour les moyennes avec un seuil à p < 0,05. Résultats Cent soixante-cinq patients ont été inclus dans l’étude et 39 patients greffés exclus. Quarante-trois (26 %) patients présentaient un taux d’IgG4 supérieur à 1350 mg/dL, 28 (65,1 %) étaient des hommes, d’âge moyen 29,7 ans et d’IMC moyen 20,6. Les caractéristiques démographiques ne différaient pas entre les groupes IgG4+ et IgG4−. Sur le plan respiratoire, il n’y avait pas de différence statistique entre les deux groupes avec un VEMS moyen à 63 % dans le groupe IgG4+ versus 62,7 % dans le groupe IgG4− ( p = 0,96). Nous n’observions pas de différence également sur la colonisation chronique ( p = 0,28) à Staphylococcus aureus , P. aeruginosa , autres germes fermentants ou mycobactéries atypiques. La prévalence de S. aureus méticilline résistant était de 18,6 % dans le groupe IgG4+ contre 9,8 % ( p = 0,17). Le nombre d’exacerbation moyen annuel dans le groupe IgG4+ est de 1,19 ± 1,4 contre 1 ± 1,35 dans le groupe témoin ( p = 0,65) ; 97,7 % patients IgG4+ présentaient une insuffisance pancréatique exocrine contre 89,3 % dans le groupe IgG4− ( p = 0,12). 16,5 % des patients présentaient un syndrome d’obstruction intestinale [ABSTRACT FROM AUTHOR]

The existence of alpha-1 antitrypsin variants with apparently unremarkable phenotypes and serum concentrations, contrasting with a clinical picture suggestive of a severe deficiency, led us to investigate whether in these cases there was a reduction or even suppression of the capacity of alpha-1 antitrypsin to inhibit elastase. To this end, in two different laboratories, we adapted and validated a method for measuring the functional activity of alpha-1 antitrypsin, based on spectrophotometric kinetic analysis of the inhibition by serum alpha-1 antitrypsin of the hydrolytic activity of porcine pancreatic elastase on a chromogenic substrate. This method has proved to be robust, reproducible and transferable and made possible to define, on the basis of an analysis of a hospital population, a functionality index with a confidence interval comprised between 0.87 and 1.2, allowing to identify subjects likely to have a functional deficiency of alpha-1 antitrypsin, whether this deficiency being of a genetic origin without any quantitative or phenotypic translation, or whether being acquired under the effect of external agents (cigarette smoke or viruses).

Introduction: Alpha1-antitrypsin deficiency is a predisposing factor for pulmonary disease and under-diagnosis is a significant problem. The results of a targeted screening in patients with respiratory symptoms possibly indicative of severe deficiency are reported here., Methods: Data were collected from March 2016 to October 2017 on patients who had a capillary blood sample collected during a consultation with a pulmonologist and sent to the laboratory for processing to determine alpha1-antitrypsin concentration, phenotype and possibly genotype., Results: In 20 months, 3728 test kits were requested by 566 pulmonologists and 718 (19 %) specimens sent: among these, 708 were analyzable and 613 were accompanied by clinical information. Of the 708 samples, 70 % had no phenotype associated with quantitative alpha1- antitrypsin deficiency, 7 % had a phenotype associated with a severe deficiency and 23 % had a phenotype associated with an intermediate deficiency. One hundred and eight patients carried at least one PI*Z allele which is considered to be a risk factor for liver disease., Conclusions: The results of this targeted screening program for alpha1- antitrypsin deficiency using a dried capillary blood sample reflect improvement in early diagnosis of this deficiency in lung disease with good adherence of the pulmonologists to this awareness campaign., (Copyright © 2020 SPLF. Published by Elsevier Masson SAS. All rights reserved.)

Background & Aims: To identify prognostic factors for liver disease in children with alpha-1 antitrypsin deficiency, irrespective of phenotype, using the DEFI-ALPHA cohort., Methods: Retrospective, then prospective from 2010, multicentre study including children known to have alpha-1 antitrypsin blood concentration below 0.8 g/L, born in France since 1989. Clinical and biological data were collected. Liver disease was classified as "severe" (portal hypertension, liver failure, liver transplantation or death); "moderate" (persistent abnormal liver biology without portal hypertension); and "mild/none" (normal or almost normal liver biology and native liver). Prognostic factors for severe liver disease were evaluated using a Cox semiparametric model., Results: In January 2017, 153 patients from 19 centres had been included; genotypes were PIZZ in 81.9%, PISZ in 8.1%, other in 10.0%. Mean ± SD follow-up was 4.7 ± 2.1 years. Half of patients had moderate liver disease. Twenty-eight children (18.3%) had severe liver disease (mean age 2.5 years, range: 0-11.6): diagnosis of alpha-1 antitrypsin deficiency was made before two months of age in 65.4%, genotypes were PIZZ in 25 (89.3%), PISZ in 2, PIM like Z in 1, 15 children underwent liver transplantation, 1 child died at 3 years of age. Neonatal cholestasis was significantly associated with severe liver disease (P = 0.007)., Conclusion: Alpha-1 antitrypsin-deficient patients presenting with neonatal cholestasis were likely to develop severe liver disease. Some patients with non-homozygous ZZ genotype can develop severe liver disease, such as PISZ and M variants, when associated with predisposing factors. Further genetic studies will help to identify other factors involved in the development of liver complications., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Alpha-1-antitrypsin deficiency (A1ATD) is a common genetic condition which predisposes to emphysema and liver disorders. It is estimated that 10-15% of homozygous individuals for the Z allele (PiZZ) may develop liver fibrosis. The optimal modalities to detect liver disease in PiZZ adult patients need to be defined. The aim of this prospective study was to perform a systematic non-invasive evaluation of the liver fibrosis by elastometry using Fibroscan ® in a cohort of A1ATD patients with emphysema., Methods: Patients followed in our respiratory unit were enrolled in this prospective study and underwent on the same day a physical examination, a biochemical profiling, an abdominal ultrasound (US) and a Fibroscan ® ., Results: Twenty-nine PiZZ adults (19 male) were included. Median age was 50.4 yrs (21.5-67.2). Median serum A1AT level was 0.20 g/L (0.15-0.33). Liver Function Tests (LFT) were not normal in 2 patients and US was abnormal in 6 patients. Two patients had both abdnormal LFT and US. Fibroscan ® was technically feasible in 28/29 (97%) patients. Median liver stiffness was 4.5 kPa (2.8-32.8), and was > 7.2 kPa in 5/28 (18%) and > 14 kPa in 2/28 (7%) patients. Liver stiffness was increased in 2/2 (100%) patients with abnormal LFT and US, in 1/4 (25%) with abnormal LFT or US and in 2/22 (10%) patients with normal LFT and US., Conclusions: Fibroscan ® is an easy and repeatable tool which can be used in PiZZ patients to screen for the presence of significant liver fibrosis and to identify patients at higher risk to develop liver complications in the future and who may benefit from a closer surveillance., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Novel anti-myeloma agents have improved patient response rates, which are historically based on reductions of the M-protein. These methods can be inaccurate for quantifying M-proteins at low concentrations. We compared the consistency and clinical impact of response assignment by electrophoretic and heavy+light chain (HLC) immunoassays post-consolidation in 463 newly diagnosed patients. The two methods gave similar assignments in patients with partial (PR; 79% agreement) or complete response (⩾CR; 92%). However, in patients achieving very good PR (VGPR) there was poor concordance between methods (45%). Median progression-free survival (PFS) for standard VGPR patients was 34.5 months; HLC responses stratified these patients further into PR, VGPR and ⩾CR, with median PFS of 21.3, 28.9 months and not reached, respectively; P<0.001. At this time, abnormal HLC ratios had better concordance with multiparametric flow cytometry (sensitivity 10 -4 ) (37 and 34% positive, respectively), compared to immunofixation (62% positive). In addition, HLC-pair suppression was identified in 38% of patients and associated with shorter PFS (30.6 months vs not reached; P<0.001). We conclude that HLC monitoring could augment electrophoretic assessments in patients achieving VGPR. The prognostic significance of HLC responses might partly depend on the patients' ability to recover their immune system, as determined by normalisation of HLC measurements.

Background & Aims: Fifteen to twenty percent of alpha-1 antitrypsin deficiency patients (A1ATD) have a severe liver outcome (portal hypertension - PHT) during childhood. Since they all share the same ZZSERPINA1 genotype and that environmental factors such as alcohol cannot be advanced, the presence of modifier genes is now well recognized. SNPs located on the SERPINA1 and MAN1B1 genes have already been tested in very few studies with contradictory or not replicated results., Methods: Our genotype-phenotype correlation study, performed on 92 ZZ children, aimed at determining once and for all if SERPINA1 and MAN1B1 polymorphisms may be implied in the onset of PHT. To do so, we also performed for the first time a complete haplotype reconstruction for data analysis., Results: The two genetic associations with severe liver disease that had been suspected previously (one SNP for SERPINA1 and another for MAN1B1) were not confirmed in our cohort. Moreover, the haplotype analysis identified only one major genetic background for the SERPINA1 Z-allele, allowing us to exclude the presence of a frequent modifier SNP within. For MAN1B1, four major haplotypes were identified but the prevalence of PHT did not significantly differ between them., Conclusion: We conclude that genetic polymorphisms in these two genes probably do not influence the onset of severe liver disease in A1ATD., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: Serum immunoglobulin (Ig) G4 elevation has been associated with several pathological conditions other than IgG4-related disease (IgG4-RD). In cystic fibrosis (CF), an elevation of specific IgG4 has been associated with colonization and infection by Pseudomonas aeruginosa. IgG4 elevation may be a marker of chronic infection or inflammatory stimulation. The aim of this study was to explore the prevalence of elevated IgG4 levels in CF and its correlation with the major clinical and microbiological features found in CF patients., Methods: In a cross-sectional study, we analyzed data from a large cohort of adult CF patients attending the CF center of Lyon University Hospital. An elevated IgG4 level was defined as being above the cut-off value of 135 mg/dL., Results: One hundred and sixty-five CF patients were analyzed. An IgG4 elevation was detected in 43 patients (26%). Compared with the control group (≤ 135 mg/dL), high IgG4 patients exhibited a greater prevalence of Staphylococcus aureus colonization and higher IgG, IgG1, IgG2 and IgE levels. No significant differences were observed in terms of pulmonary function, colonization with Pseudomonas aeruginosa, or the annual rate of bronchial exacerbations., Conclusion: An elevated IgG4 serum level was frequently detected in adult CF patients and did not appear to be associated with poor lung function. We suggest that IgG4 elevation is a marker of the activation of tolerance. Its clinical significance remains to be demonstrated.

Background: Alpha 1 antitrypsin (A1AT) deficiency (A1ATD) is potentially associated with a high degree of liver and/or lung disease. Apart from the most frequent deficiency alleles, Pi S and Pi Z, some A1AT alleles of clinical significance may be easily misdiagnosed. This is typically the case of the Pi Mmalton variant which shares the same 'gain-of-function' liver toxicity than Pi Z and the same 'loss of function' lung disease as well., Methods: The biological diagnosis of A1ATD typically relies on a low serum concentration associated with an abnormal isoelectric focusing (IEF) pattern of migration. However, Sanger direct DNA sequencing may be required for deficiency alleles without biochemical expression (Null alleles) or for A1AT variants whose IEF profiles resemble the wild-type and sub-types M allele but with a low concentration., Results: We report four cases of A1ATD involving the deficient Pi Mmalton allele with very different clinical expressions: (i) one Mmalton/Mmalton with liver fibrosis and cirrhosis, (ii) two Mmalton/Z with chronic pulmonary obstructive disease in one case and (iii) one M/Mmalton without liver or lung disease. In both cases, the correct diagnosis has necessitated a genetic analysis., Conclusions: Our study provides another example of Pi Mmalton homozygosity associated with a severe liver disease that emphasizes the necessity of a not delayed diagnosis. The great clinical heterogeneity of the other genotypes (which is in agreement with the literature data) questions about the role of environmental and other modifier genes in the pathogenicity of A1ATD.

Introduction: The alpha-1 antitrypsin (α1-AT) deficiency, most frequently caused by homozygosity for the Z variant (SERPINA1: c.1096 G>A; Glu342Lys), can give rise to two clinical patterns: (i) respiratory impairment with emphysema (mainly in adulthood) because of a pulmonary quantitative defect in anti-elastase activity; (ii) hepatic impairment (mainly in childhood) due to the misfolding of the PiZ protein which accumulates in hepatocytes thus providing cytotoxicity., Current Knowledge: To date, the clinical and genetic factors responsible for the development of major hepatic injuries (fibrosis and portal hypertension) during childhood in PiZ patients are not known., Methods: The DEFI-ALPHA cohort, created in 2008, aims to inventory and prospectively study all α1-AT deficient children diagnosed and included after occurrence of a hepatic sign. The POLYGEN DEFI-ALPHA PHRC has recently (2013) been added to the project to identify modifiers genes by two complementary approaches: (i) the candidate genes strategy with the SERPINA1, CFTR (cystic fibrosis gene), MAN1B1 and SORL1 genes, these two latter being implied in the degradation of misfolding proteins; (ii) the whole exome sequencing (WES) strategy in families in which the PiZ proband has a PiZ brother or sister free of any hepatic sign., Expected Results: The clinical parameter we want to explain is the apparition of a portal hypertension in PiZ children. In the DEFI-ALPHA project, three criteria will be tested: (i) age of inclusion in the cohort, (ii) the way of inclusion (neo-natal icterus or later hepatic impairment) and (iii) treatment or not with ursodesoxycholic acid and, if so, its duration. Genetically, polymorphisms on the SERPINA1 and MAN1B1 genes have already been associated in the literature with different clinical evolutions of the A1ATD but very inconstantly. Our study thus aims to confirm or not this association. The CFTR and SORL1 genes have never been studied in the α1-AT deficiency. Finally, the whole exome sequencing strategy could allow the discovery of new unexpected modifiers genes in this disease., (Copyright © 2015 SPLF. Published by Elsevier Masson SAS. All rights reserved.)

Alpha- 1-antitrypsin (A1AT) deficiency is a hereditary autosomal codominant genetic disorder resulting in low circulating levels of A1AT and leading to lung and/or liver disease. It remains underdiagnosed and only 5 to 10% of PIZZ patients, the most common form of severe A1AT deficiency, would be actually identified in France. Facilitating early diagnosis of A1AT deficiency would allow a better management of this disease; therefore we have developed and standardized in three laboratories involved in this study, a diagnostic test on dried blood spots (DBS) including quantitative A1AT measurement, phenotyping by IEF electrophoresis and, if necessary, genotyping by SERPINA1 gene sequencing. We performed a quantitative assay on 90 DBS samples by immunoturbidimetric or immunonephelometric methods. We demonstrated that both methods were suitable for this type of sampling and the results obtained were highly correlated (R(2)>0.9) between the three laboratories: for a target value of 1.00 g/L, the results obtained from the three laboratories were between 1.00 and 1.02 g/L. Phenotyping and genotyping were performed under redefined operating conditions and adapted to the analysis of DBS samples. The results were comparable with those obtained for venous blood samples. Following this work, it becomes possible to provide pulmonologists with a reliable kit to perform a capillary blood sampling on filter paper which would allow a large-scale screening of A1AT deficiency in the population particularly affected by this genetic condition.

Unlabelled: Renal dysfunction is frequent in liver cirrhosis and is a strong prognostic predictor of orthotopic liver transplantation (OLT) outcome. Therefore, an accurate evaluation of the glomerular filtration rate (GFR) is crucial in pre-OLT patients. However, in these patients plasma creatinine (Pcr) is inaccurate and the place of serum cystatine C (CystC) is still debated. New GFR-predicting equations, based on standardized assays of Pcr and/or CystC, have been recently recommended in the general population but their performance in cirrhosis patients has been rarely studied. We evaluated the performance of the recently published Chronic Kidney Disease Epidemiology Collaboration equations (CKD-EPI-Pcr, CKD-EPI-CystC, and CKD-EPI-Pcr-CystC) and the more classical ones (4- and 6-variable MDRD and Hoek formulas) in cirrhosis patients referred for renal evaluation before OLT. Inulin clearance was performed in 202 consecutive patients together with the determination of Pcr and CystC with assays traceable to primary reference materials. The performance of the GFR-predicting equations was evaluated according to ascites severity (no, moderate, or refractory) and to hepatic and renal dysfunctions (MELD score  ≤ or >15 and KDOQI stages, respectively). In the whole population, CystC-based equations showed a better performance than Pcr-based ones (lower bias and higher 10% and 30% accuracies). CKD-EPI-CystC equation showed the best performance whatever the ascites severity and in presence of a significant renal dysfunction (GFR <60 mL/min/1.73 m(2))., Conclusion: Pcr-based GFR predicting equations are not reliable in pre-OLT patients even when an IDMS-traceable enzymatic Pcr assay is used. Whenever a CystC-assay traceable to primary reference materials is performed and when a true measurement of GFR is not possible, CystC-based equations, especially CKD-EPI-CystC, may be recommended to evaluate renal function and for KDOQI staging., (© 2013 by the American Association for the Study of Liver Diseases.)

The diagnosis of deficiency of alpha-1 antitrypsin (A1AT) is based on isoelectric focusing of serum proteins and the extent of serum. However, the focusing is technically difficult and a greatly reduced concentration in abnormal A1AT tapeless does not differentiate an unstable variant of a variant called 'null' (that is to say without any phenotypic expression) to 'heterozygous' state. In this study, we compared the results of the assay, the phenotype and genotype of A1AT in 50 patients. Normal A1AT alleles (Pi*M1 to Pi*M4) or loss of the most common (Pi*S and Pi*Z) were clearly identified in phenotyping. However, genotyping was necessary to characterize: (i) certain alleles rarer A1AT (S-Munich, X-Christchurch); (ii) a null allele and; (iii) two new alleles A1AT not yet described in the literature. In conclusion, although the A1AT genotyping is generally not necessary, it is necessary to resolve complex cases and to obtain witnesses validated for isoelectric focusing.

Background: It has been reported that first morning specimens are more reliable than random spot specimens to assess 24-hour urinary albumin excretion rate (UAER), especially if albuminuria is expressed as albuminuria to creatininuria ratio. We aimed to investigate the influence of (a) posture and activity and (b) the units to best estimate 24-hour albuminuria., Methods: In this retrospective study, 24-hour UAER was compared to 60 min 'supine' and 90 min 'activity' albuminuria in 124 patients tested for resistant hypertension. The ability to adjust urinary albumin concentration (UAC) to creatininuria (ACR) or to collection duration (tAER) values in order to increase the reliability of albuminuria values was also analyzed., Results: Compared to 24-hour UAER, UAC (mg/l), tAER (μg/min) and ACR (mg/mmol) during the supine period had a similar concordance rate in normo-, micro- and macroalbuminuric patients. The UAC in the supine period was well related to 24-hour UAER. However, UAC almost doubled during activity. Adjustment to creatininuria improved the correlation between albuminuria during both periods and 24-hour UAER, but mainly during the activity period., Conclusions: Our results confirm that UAC is dependent on physical activity. Correction of UAC by creatininuria (ACR) provides a satisfactory estimation of 24-hour UAER. Thus, for practical reasons, it is advisable to use ACR, where no differences appear to exist, whether a supine urine sample or an activity urine sample is obtained., (Copyright © 2010 S. Karger AG, Basel.)

Objective: Cast nephropathy, due to free light chain (FLC) toxicity, is the main cause of acute kidney injury in multiple myeloma, with about 10% of patients requiring dialysis. In these patients, in addition to chemotherapy that prevents FLC production, daily hemodialysis using high cutoff or adsorptive membranes, showed promising results by decreasing quickly toxic serum FLC concentrations., Case History: We report here the case of 2 patients presenting with acute kidney injury and high FLC serum concentration and M-components one with IgG Kappa and the other with IgD lambda. Both were treated with bortezomib and dexamethasone and received a 24-h continuous hemodialysis using a high and sharp cutoff (around 35,000 Daltons) polysulfone membrane (ultraflux® HD 1000, Fresenius Medical Care GmbH, Bad Homburg, Germany) with citrate regional anticoagulation using a safe and dedicated device (multi filtrate Ci-Ca®)., Conclusion: Despite similar range of depuration, serum plasma FLC decreased importantly in the patient with the kappa type who recovered but was unchanged in the lambda type patient who remained under maintenance dialysis. Further studies are needed to confirm this new approach therapy.

We sought a possible correlation between the clinical manifestations of staphylococcal scalded-skin syndrome (SSSS) and the serotype of exfoliative toxins (ET) by PCR screening of the eta and etb genes in Staphylococcus aureus strains isolated from 103 patients with generalized SSSS and 95 patients with bullous impetigo. The eta gene and the etb gene were detected in, respectively, 31 (30%) and 20 (19%) episodes of generalized SSSS and 57 (60%) and 5 (5%) episodes of bullous impetigo. Both genes were detected in 52 (50%) episodes of generalized SSS and 33 (35%) episodes of bullous impetigo. To explain this link between etb and generalized SSSS, we examined the distribution of ETA- and ETB-specific antibodies in the healthy population (n = 175) and found that the anti-ETB antibody titer was lower than the anti-ETA titer. Thus, ETA is associated with bullous impetigo and ETB is associated with generalized SSSS, possibly owing to a lower titer of anti-ETB neutralizing antibodies in the general population.

Since the introduction of fully automated nephelometric systems simultaneous measurements of immunoglobulin light chains kappa (kappa) and lambda (lambda) and IgG, IgA and IgM have become increasingly used for the routine assessment of humoral immunity. From these data two ratios were calculated, the kappa/lambda ratio and the heavy chains to light chains ratio. As changes in these ratios might have some predictive clinical value besides reflecting a monoclonal component, it is necessary to know mean and reference limits of these ratios. On account of differences in the calibration method of the light chains measurements (either free light chains or light chains bound to a complete molecule) and of differences in the calculation method of the heavy chains to light chains ratio we were led to conduct our own investigation. IgG, IgA and IgM and kappa and lambda light chains were immunonephelometrically measured in the sera of 84 blood donors. For each sample theoretical values for kappa + lambda, kappa and lambda, and kappa/lambda were calculated using the existing relation between the concentration of a given immunoglobulin and the concentration of bound light chains. Using the Valtec Protocole and the t test we were able to evidence highly significant differences (p < 10(-4) between theoretical and experimental values of kappa, lambda and kappa + lambda; those differences could be proved to be directly linked to the nephelometric technique itself. However the experimental kappa/lambda ratio did not appear to differ from the theoretical one nor the standardization method to have an effect on the reference values of this ratio, our values (mean and reference limits, 1.81, 1.29-2.53) being very similar to previously published results. Concerning the so called heavy chains to light chains ratio two methods were used to express it, one consisting in the ratio of the theoretical kappa + lambda value to the experimental one with the following results, 1.05 and 0.93-1.18 for the mean and reference limits and the other one using the raw data. The results were as follows: mean 3.50, reference limits 3.11-3.94.

Purpose: We investigated the place of direct plasma cell proliferation analysis beside other biologic data in monoclonal gammopathy, particularly the serum level of C-reactive protein (C-RP) PATIENTS: Eighty patients were studied at the time of their diagnosis. Patients with a serum creatinine level greater than 200 mumol/L were excluded., Methods: Plasma cell proliferation analysis was performed after bromodeoxyuridine incorporation and double immunoenzymatic labeling on cytological smears, making determination of the plasma cell labeling index (LI) possible. The other biologic variables studied were related to tumor burden (plasmacytosis, hemoglobin, serum levels of monoclonal immunoglobulin, albumin) or to kidney function (creatinine). beta 2-microglobulin (beta 2-M), C-RP, lactic dehydrogenase (LDH), and calcemia were also assessed., Results: No correlation was found between LI and serum C-RP. LDH was the sole variable significantly correlated to C-RP (P < 0.01). Besides the biologic parameters used for the staging according to the Durie and Salmon classification, beta 2-M, albumin and LI were significantly different between stages (P < 0.0002, < 0.0004, < 0.00001). LDH and C-RP showed no significant difference. Results were similar when patients with and without bone lesions were analyzed separately. Multivariate analysis ranked these variables as follows with respect to prognostic value: beta 2-M, LI, and age., Conclusion: Among the variables analyzed, LI is the sole true reflector of cell proliferation. We confirm its diagnostic and prognostic value.

Among 998 children with recurrent respiratory diseases 26 children with selective IgA deficiency were found. Three groups were considered according to IgA level in serum: group I with IgA under 0.05 g per litre; group II with IgA between 0.05 and 0.3 g per litre; group III with IgA above 0.3 and under 1 g per litre. Non specific immunity was studied in these patients including immunoglobulin levels, alpha-1-antitrypsin (A.A.T.) phenotypes, phagocytosis of staphylococcus aureus by PMN, lysozyme level, complement system. Cellular immunity was evaluated by IDR tests and rosette forming cells (RE). Only non specific immune systems were disturbed in some patients and appeared as aggravating factors in IgA deficient patients. We found: Abnormal phenotypes of ATT in 11 cases; deficiencies of engulfment in 6 cases, of bactericidal activities of PMN in 7 cases out of 16 studied; decrease of lysozyme level in 4 cases out of 17 studied; increase of IgE level in 9 cases with atopic symptoms in 7 patients. In our experience the chief aggravating factor in IgA deficient patients is abnormal phenotype of AAT.

Group-specific component (vitamin D-binding protein) was purified to homogeneity from human plasma by a three-step procedure involving pseudo-ligand affinity chromatography on immobilized Cibacron blue F3-GA followed by gel filtration and ion-exchange chromatography. Upon pseudo-ligand chromatography, Gc globulin was separated into two peaks. The first, which represented approx. 4% of the total Gc globulin, was eluted together with other alpha-globulins of similar Mr and/or pI, and the second (96% of Gc globulin) was clearly retarded. Collection of the latter provided a fraction 10-fold enriched in Gc globulin, with yields higher than 90%. Incubation of plasma with trace amounts of radioactively-labeled 25-OH vitamin D3 showed that the radioactivity coeluted with the first peak. In addition, after saturation with 25-OH vitamin D3, all the Gc globulin was eluted in the first peak. This indicates that the two peaks correspond to the holo and the apo forms of the protein, respectively, and suggests that either the interaction of the apo form with the Cibacron blue dye involves the binding site for vitamin D metabolites, or that the holo-protein undergoes a conformational change as a consequence of formation of the complex.

The microheterogeneity of the I allele of the Pi system of APi (alpha-1-antitrypsin) was studied in 43 individuals with the new PAGIF technique. The unique aspect of the I allele product (unequal distribution of bands 4 and 6), previously demonstrated with acid-starch gel, was confirmed. In addition, two subtypes of the Pi I allele--I1 and I2--were clearly distinguished. Serum concentrations of APi associated with the expression of the I allele were significantly decreased (68% of normal values) and thus very similar to those associated with the expression of the S allele. This indicates that the I allele can be considered as a "deficient" allele of the Pi system.