PI and OPG/RANKL levels in human osteoblast cells

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Spectrum of mutations in Italian patients with familial hypercholesterolemia: New results from the LIPIGEN study

Background Familial hypercholesterolemia (FH) is an autosomal dominant disease characterized by elevated plasma levels of LDL-cholesterol that confers an increased risk of premature atherosclerotic cardiovascular disease. Early identification and treatment of FH patients can improve prognosis and reduce the burden of cardiovascular mortality. Aim of this study was to perform the mutational analysis of FH patients identified through a collaboration of 20 Lipid Clinics in Italy (LIPIGEN Study). Methods We recruited 1592 individuals with a clinical diagnosis of definite or probable FH according to the Dutch Lipid Clinic Network criteria. We performed a parallel sequencing of the major candidate genes for monogenic hypercholesterolemia (LDLR, APOB, PCSK9, APOE, LDLRAP1, STAP1). Results A total of 213 variants were detected in 1076 subjects. About 90% of them had a pathogenic or likely pathogenic variants. More than 94% of patients carried pathogenic variants in LDLR gene, 27 of which were novel. Pathogenic variants in APOB and PCSK9 were exceedingly rare. We found 4 true homozygotes and 5 putative compound heterozygotes for pathogenic variants in LDLR gene, as well as 5 double heterozygotes for LDLR/APOB pathogenic variants. Two patients were homozygous for pathogenic variants in LDLRAP1 gene resulting in autosomal recessive hypercholesterolemia. One patient was found to be heterozygous for the ApoE variant p.(Leu167del), known to confer an FH phenotype. Conclusions This study shows the molecular characteristics of the FH patients identified in Italy over the last two years. Full phenotypic characterization of these patients and cascade screening of family members is now in progress

Familial hypercholesterolemia: The Italian Atherosclerosis Society Network (LIPIGEN)

BACKGROUND AND AIMS: Primary dyslipidemias are a heterogeneous group of disorders characterized by abnormal levels of circulating lipoproteins. Among them, familial hypercholesterolemia is the most common lipid disorder that predisposes for premature cardiovascular disease. We set up an Italian nationwide network aimed at facilitating the clinical and genetic diagnosis of genetic dyslipidemias named LIPIGEN (LIpid TransPort Disorders Italian GEnetic Network). METHODS: Observational, multicenter, retrospective and prospective study involving about 40 Italian clinical centers. Genetic testing of the appropriate candidate genes at one of six molecular diagnostic laboratories serving as nationwide DNA diagnostic centers. RESULTS AND CONCLUSIONS: From 2012 to October 2016, available biochemical and clinical information of 3480 subjects with familial hypercholesterolemia identified according to the Dutch Lipid Clinic Network (DLCN) score were included in the database and genetic analysis was performed in 97.8% of subjects, with a mutation detection rate of 92.0% in patients with DLCN score 656. The establishment of the LIPIGEN network will have important effects on clinical management and it will improve the overall identification and treatment of primary dyslipidemias in Italy

Semi-Continuous Adsorption Processes with Multi-Walled Carbon Nanotubes for the Treatment of Water Contaminated by an Organic Textile Dye

Adsorbent columns, containing different amounts of multi-walled carbon nanotubes (MWCNTs), in a semicontinuous process were studied. The optimal conditions for the discoloration of water contaminated by an azoic organic textile dye were investigated. In particular, as representative of contaminated water, a highly concentrated solution of Reactive Black 5 (RB5) equal to 37 mg/L was utilized. A predetermined volume of dye solution, equal to 100 mL, was subjected to repeated cycles of adsorption until the eluted solution became colorless. This adsorption operation was carried out for different types of columns. Adsorbent performances as a function of characteristics of each column were investigated. For each column, the optimum quantity of MWCNTs, maximum volume of treatable solution, carbon usage rate (CUR), empty bed contact time (EBCT), and adsorption capacity were determined. The permeate was characterized by UV-VIS analysis and TOC analysis, while adsorbent material (MWCNTs) was characterized by thermogravimetric TG-DTA analysis. The column containing 2.5 g of carbon nanotubes was revealed to be the best one for the total amount of Reactive Black 5 adsorbed, i.e., 55 mg/g(MWCNTs) The research has shown the high adsorption efficiency of carbon nanotubes toward RB5 dye, highlighting the degradation of the dye molecule and the stratification, inside the columns, of the adsorbed compound

A multicenter evaluation of the Abbott RealTime HCV genotype II assay

Viral genotype is an important determinant of the therapeutical outcome of the chronic hepatitis C and is useful in clinical practice to determine the duration of treatment1.While the viral type shows a clear association with therapeutic success, there is currently no evidence to that effect for HCV subtype, whose value is thus confined to epidemiological studies.The Abbott RealTime HCV Genotype II assay, through the use of Minor Groove Binder probes (MGB) is able to distinguish genotypes 1 to 6 (target 5’-UTR region) and subtypes 1a and 1b (NS5B region). In four different Italian centers a comparison between the Abbott RealTime HCV Genotype II assay and the Versant HCV Genotype 2.0 (LIPA) has been performed.A total of 143 non selected samples with the request of HCV genotyping have been analysed. 141/143 samples (98.6%) have provided reportable results with both tests (2 indeterminates with LIPA). Concordance at the type level was 96.5% (136/141). Considering the 136 concordant samples, the distribution was as follows: type 1 = 61 (44.9%), 2 = 36 (26.5%), 3 = 21 (15.4%), 4 = 17 (12.5% ) 5 = 1 (0.7%). Both assays assigned subtype in 56/61 (91.8%) samples of genotype 1 (3 and 2 samples only provided the type for LIPA and Abbott, respectively) and 50/56 (89.3%) had concordant subtype. It is worth to note that 4 of the 5 samples with discordant subtype Abbott 1a/LiPA 1b came from the only center that used for LIPA the 5-UTR amplicon, loosing the benefit of the core region which has been introduced in the version 2 of the test to improve the accuracy in distinguishing between 1a and 1b.There was only one discordant sample at type level (Abbott 4, LIPA 1b) which after sequencing and phylogenetic analysis was resolved as type 4. Four mixed infections were detected, 3 with the Abbott test (two 1a+4 and one 1b+3) and 1 with the LIPA test (1a+3). In all cases the comparison test showed a single genotype 1 infection. The new Abbott RealTime HCV Genotype II assay showed a high correlation with the Versant HCV Genotype 2.0 assay (LIPA).The automation platform m2000 system (Abbott), together with objective interpretation and digital archiving results may be particularly advantageous for the laboratory

Standardization of nucleic acid amplification tests in diagnostic molecular microbiology

Nucleic acid detection by amplification (NAT) assays began to be used in diagnostic microbiology about twenty years ago. Since then, the progress of molecular methods has been continuous and rapid, aimed to obtain more and more sensitive and specific results and automation.To actually achieve such objectives, microbiology laboratories have to use suited quality controls and to participate to external quality control programs. Quality controls for NAT assays need to be adjusted to follow the evolution of the molecular methods.The need of standardization and suited controls for NAT assays became evident approximately by the middle of ’90 years.Thus International Standards (IS) were established by the World Health Organization, for the main blood borne viruses. Such preparations had a well known concentration expressed as International Units, a new measure unit for nucleic acid quantitation. Then, IS were used in order to calibrate working reagents and synthetic calibrators. Another tool for standardization are multicentre studies aimed to verify the performances of participating laboratories using different molecular methods, and allow the comparison of inter-laboratory results.The CoSBio of AMCLI, as other European organizations, in the last years promoted, some multicentre studies on NAT assays. The results of such studies demonstrated a good level of diagnostic performances of participating laboratories. However, with the introduction of the real time PCR (RQ-PCR) a new problem emerged: the variability of results obtained by molecular methods due to different coefficient of linear amplification. Solutions suggested by Literature to this and similar problems are examined, mainly focusing on the assessment of the Calibration curves in the RQ-PCR. Finally, the Authors signal some important fields in the molecular diagnostics, such as the detection of HPV and the detection of BKV, still lacking of sufficient standardization