Rossella Zenobi

Atherosclerosis, dyslipidemia, and inflammation: the significant role of polyunsaturated Fatty acids.

Phospholipids play an essential role in cell membrane structure and function. The length and number of double bonds of fatty acids in membrane phospholipids are main determinants of fluidity, transport systems, activity of membrane-bound enzymes, and susceptibility to lipid peroxidation. The fatty acid profile of serum lipids, especially the phospholipids, reflects the fatty acid composition of cell membranes. Moreover, long-chain n-3 polyunsatured fatty acids decrease very-low-density lipoprotein assembly and secretion reducing triacylglycerol production. N-6 and n-3 polyunsatured fatty acids are the precursors of signalling molecules, termed “eicosanoids,” which play an important role in the regulation of inflammation. Eicosanoids derived from n-6 polyunsatured fatty acids have proinflammatory actions, while eicosanoids derived from n-3 polyunsatured fatty acids have anti-inflammatory ones. Previous studies showed that inflammation contributes to both the onset and progression of atherosclerosis: actually, atherosclerosis is predominantly a chronic low-grade inflammatory disease of the vessel wall. Several studies suggested the relationship between long-chain n-3 polyunsaturated fatty acids and inflammation, showing that fatty acids may decrease endothelial activation and affect eicosanoid metabolism.

Human endogenous retroviruses and ADHD

Abstract Objectives. Several lines of evidences suggest that human endogenous retroviruses (HERVs) are implicated in the development of many complex diseases with a multifactorial aetiology and a strong heritability, such as neurological and psychiatric diseases. Attention deficit hyperactivity Disorder (ADHD) is a neurodevelopmental disorder that results from a complex interaction of environmental, biological and genetic factors. Our aim was to analyse the expression levels of three HERV families (HERV-H, K and W) in patients with ADHD. Methods. The expression of retroviral mRNAs from the three HERV families was evaluated in peripheral blood mononuclear cells (PBMCs) from 30 patients with ADHD and 30 healthy controls by quantitative RT-PCR. Results. The expression levels of HERV-H are significantly higher in patients with ADHD compared to healthy controls, while there are no differences in the expression levels of HERV-K and W. Conclusions. Since the ADHD aetiology is due to a complex interaction of environmental, biological and genetic factors, HERVs may represent one link among these factors and clinical phenotype of ADHD. A future confirmation of HERV-H overexpression in a larger number of ADHD patients will make possible to identify it as a new parameter for this clinical condition, also contributing to deepen the study on the role of HERVs in the neurodevelopment diseases.

Transcriptional Activity of Human Endogenous Retroviruses in Human Peripheral Blood Mononuclear Cells

Human endogenous retroviruses (HERVs) have been implicated in human physiology and in human pathology. A better knowledge of the retroviral transcriptional activity in the general population and during the life span would greatly help the debate on its pathologic potential. The transcriptional activity of four HERV families (H, K, W, and E) was assessed, by qualitative and quantitative PCR, in PBMCs from 261 individuals aged from 1 to 80 years. Our results show that HERV-H, HERV-K, and HERV-W, but not HERV-E, are transcriptionally active in the test population already in the early childhood. In addition, the transcriptional levels of HERV-H, HERV-K, and HERV-W change significantly during the life span, albeit with distinct patterns. Our results, reinforce the hypothesis of a physiological correlation between HERVs activity and the different stages of life in humans. Studies aiming at identifying the factors, which are responsible for these changes during the individuals life, are still needed. Although the observed phenomena are presumably subjected to great variability, the basal transcriptional activity of each individual, also depending on the different ages of life, must be carefully considered in all the studies involving HERVs as causative agents of disease.

Minimal tumour burden in haematological diseases: a step forward with quantitative assessment of Bence‐Jones in nephelometry?

Serum and urine free light chains (FLCs) monoclonal immunoglobulin (k and k) are important markers in the diagnosis and monitoring of B cell proliferative disorders, such as Multiple Myeloma (MM), Monoclonal Gammopathy (MG), Monoclonal Gammopathy of Undetermined Significance (MGUS), Amyloidosis (AL) and related disorders (Bird et al, 2009; Kyle et al, 2010). MM is a progressive neoplastic disease characterized by bone marrow plasmacytosis (plasma cell tumour) and overproduction of an intact monoclonal immunoglobulin (IgG, IgA, IgM, IgD or IgE) and/or Bence Jones protein. The presence of one of the three following criteria allows the diagnosis of myeloma in affected patients: layers or clusters of plasma cells in the bone, osteolytic lesions (without the presence of metastatic cancer or granulomatous disease) or Bence Jones proteinuria (Durie et al, 2006). Determination of the Bence Jones protein (BJP) is essential when clinical suspicion of MG persists and serum tests are negative; moreover it is useful to establish a diagnosis of MG (Bird et al, 2009; Dimopoulos et al, 2011) and to monitor response to therapy in MM. It has been suggested that the FLCs assay can replace the traditional 24-h urine protein electrophoresis and immunofixation as part of a diagnostic screen (International Myeloma Working Group 2003; Eslick & Talaulikar, 2013). The excess production of FLCs (over 10–30 g/day) by a neoplastic clone of plasma cells saturates their absorption in the proximal renal tubules, causing damage and proteinuria, with large amounts of FLCs detectable in the urine. Urine FLCs (BJP) analysis can probably still be useful for initiating therapy and to highlight minimal tumour burden in patients with MM (Eslick & Talaulikar, 2013; Paiva et al, 2015a) and related disorders. There is growing interest in Minimal Residual Disease (MRD) monitoring, which can be used as a prognostic factor and to predict patients’ outcomes in MM (Paiva et al, 2015b). Guidelines recommend that immunofixation (IFE) of urine should be used to detect BJP (Durie et al, 2006; Bird et al, 2009). Currently, BJP quantification must be carried out on spot urine sample by densitometry of the corresponding immunoelectrophoretic band. This method is complicated by the different affinity for used dyes, the coincidental migration of other proteins and is operator dependent; therefore, quantitative analysis could be performed by nephelometric/turbidimetric assay. Although IFE is the ‘gold standard’ for the determination of BJP, a new nephelometric method, based on monoclonal antibodies, has been developed (Pretorius et al, 2012). The monoclonal antibodies provide a more accurate measurement reducing a possible overestimation when polyclonal antibodies are used. The aim of the present study was to compare the performance of nephelometric assay to IFE agarose gel in urine samples and to assess the correlation between the results of these two tests and the clinical diagnosis (negative or positive for B cell proliferative disorders). Serum and urine samples were collected from 378 patients (186 female and 192 male; one sample for each patient: no follow-up sample needed for this type of analysis) who presented to “Tor Vergata University Hospital”. Patients were evaluated with serum electrophoresis (SPE), urine immunofixation (IFE) and nephelometric assays for urine FLCs and additional routine analyses. Exclusion criteria were history and/or signs of chronic disease as primary renal failure or patients undergoing dialysis. FLCs measurement was performed using the N Latex FLC kit based on a mixture of monoclonal antibodies for use on the BN ProSpec System analyser (Siemens GmbH, Marburg, Germany) (Pretorius et al, 2012). BJP was determined by immunofixation on a semi-automated agarose electrophoresis system (Hydragel 4 BJ and Hydrasys; Sebia, Florence, Italy). The gels were evaluated for the presence of FLCs by three independent operators and reported as positive or negative. The evaluation was performed without knowledge of the outcome from the nephelometric assays. Data obtained show a significant correlation between nephelometric and IFE assays (Spearman correlation r = 0 82, P < 0 001). We analysed the Receiver-Operator Characteristic (ROC) curves for kappa and lambda nephelometric FLCs assay with respect to diagnosis (Fig 1A and B). The areas under ROC curves were 1 for both kappa and lambda BJP. We found 99% and 100% sensitivity and 100% and 99% specificity, for kappa and lambda FLCs, respectively, and the optimal cutoff for kappa FLCs was 0 25 mg/l and 0 11 g/l for lambda FLCs. We made a new correlation of the nephelometric data with respect to diagnosis that was re-assessed and shown to be more significant (r = 0 98, P < 0 001; Spearman correlacorrespondence

Multiple Sclerosis: kFLC index values related to gender

BACKGROUND Multiple sclerosis (MS) is a chronic multifactorial inflammatory and neurodegenerative disease of the central nervous system (CNS). The identification of biomarkers with good diagnostic and prognostic power is of great importance for monitoring and treating MS patients. METHODS We analyzed serum and cerebrospinal fluid of 228 patients, with different neurological disorders and with MS to confirm our previous results and determine a possible gender difference of kFLC Index cut-off. RESULTS We have obtained a kFLC Index cut-off of 12.5 (100% specificity and 90.4% sensitivity) and 11 (100% specificity and 97.5% sensitivity) for women and men with MS respectively. CONCLUSIONS This study reinforces the importance that kFLC Index could have as a diagnostic aid to detect MS. Our data highlight a difference in the cut-off of the kFLC Index calculated by gender; male patients with a kFLC Index value greater than 11 are at higher risk to develop MS respect females having the same result.

Reference intervals for HbA1c partitioned for gender and age: a multicenter study.

Most recent estimates indicate that 8.8 % of adults (415 million people) have diabetes, and the number of people with the disease is set to rise beyond 642 million in \25 years. This disease is often asymptomatic in its early stages and can remain undetected for several years [1], thus causing the onset of micro-vascular and macro-vascular complications, such as retinopathy, kidney failure, limb amputation and cardiovascular diseases [2], which are the most common cause of death and disability among people with diabetes. In order to minimize the risk of these complications and associated healthcare costs, the early detection and management of diabetes and prediabetes is imperative [1]. According to the WHO, glycated haemoglobin (HbA1c) values between 6.0 and 6.5 % (42–47 mmol/mol) indicate a high risk for the onset of diabetes. Moreover, the International Expert Committee (IEC) recommends that subjects with an HbA1c value within this range should be given interventions; while the American Diabetes Association (ADA) recommends lower HbA1c levels (5.6–6.5 %; 38–47 mmol/mol), together with other tests, to define prediabetes. We currently use the reference ranges recommended by the ADA, but these may be unsuitable for the European population. Furthermore, we recently showed that there is a significant difference in mean HbA1c values measured in healthy donors based on gender [3]. In this context, it seemed interesting to investigate whether there are significant differences in HbA1c values measured in non-diabetic donors not only by gender but also by age.