Biochimica Clinica: VOL.45 N.3

Overexpression of cyclin D1 (CCND1) occurs often in tumoral plasma cells, mainly – but not exclusively – as a result of the presence of the t(11;14) translocation. Of note, CCND1 mRNA overexpression or the presence of the t(11;14) translocation was shown to impact on the response to anti-plasma cell drugs and influence prognosis, both in patients with multiple myeloma and with immunoglobulin light chain (AL) amyloidosis. In this study, we performed a systematic revision of published literature on molecular assays to measure CCND1 transcript levels in plasma cell dyscrasias, in order to describe currently available assays, their technical characteristics and main applications. Relevant scientific articles were search on PubMed as of October 2020 using combinations of appropriate key words. Of 165 unique studies retrieved, 11 articles fulfilled the inclusion criteria and were further analyzed. Overall, 8 different molecular assays were described and characterized. Most of the studies focused on multiple myeloma, with some studies including also MGUS, plasma cell leukemia and/or AL amyloidosis. Assay design, technical validation and field of application of each assay were systematically reviewed. As more knowledge is gained about the impact of CCND1 expression levels on the biology of tumoral plasma cells and their response to anti-plasma cell drugs, including novel agents specifically targeting t(11;14)-positive clones, molecular assays to quantify CCND1 expression levels in tumoral plasma cells may become a useful complementation to molecular cytogenetics, towards a precision medicine approach to diagnose and treat plasma cell disorders which is based on laboratory medicine.

Introduction: in the last decade, cancer immunotherapy has delivered impressive results in clinical settings. However, its efficacy has not been consistent probably because of several environmental and genetic factors influencing the outcome. Many studies have indicated that intestinal microbiota could affect the outcome of immune checkpoint inhibitors-based immunotherapy, both in animal models and patients. In particular, the Bifidobacterium genus seems to have a role as a positive regulator of in vivo antitumor immunity by promoting proinflammatory signals in innate immune cells. According to the considerable evidence that demonstrated its crucial role in the carcinogenesis and, overall, in the response to immunotherapy, we decided to use a commercial probiotic and grow its principal strain, the Bifidobacterium longum BB-536, in order to test its capability to affect antitumoral immune responses.
Methods: prior to in vivo studies, we carried out a feasibility evaluation study to test in vitro, antitumoral effects of the isolated probiotic strain. Tumor cell viability was used as parameter to determine Bifidobacterium longum BB-536 anti-proliferative ability before or after heat inactivation.
Results: interestingly, we found that B. longum inhibits cell growth, both in mouse melanoma B16-OVA and colorectal CT26 cells, showing a more pronounced effect on the latter ones.
Conclusion: this preliminary evaluation of live and heat-inactivated probiotic in tumor cell lines indicates a potential cell growth inhibitory effect of these bacterial strains and encourage further studies in mouse models.

Introduction: glucose measurement is pivotal in the management of subjects with diabetes mellitus. Laboratories should provide the most useful information to the clinician in order to ensure the best patient outcome.
Methods: in October 2019 a survey has been conducted by SIBioC and SIPMeL Study Groups on “Diabetes Mellitus” and by the Italian Diabetes Society to verify if their recommendations on preanalytical phase have had an impact on Italian laboratory procedures. Fifteen questions were submitted to all SIBioC and SIPMeL members and 190 complete responses were collected, corresponding to around 5% of all Italian laboratories.
Results: 74% of the laboratories (n=46) are aware of the recommendations of the “Diabetes Mellitus” Study Groups, but only 24% apply them. 61% of the first group centrifuges the collection tubes immediately, providing a rapid plasma separation; 9% place the tubes immediately in an ice-water slurry and separate the plasma within 30 minutes. Only 14 of the responders use citrate buffer/sodium EDTA/sodium fluoride (NaF) tube in lyophilic formulation.
The survey allowed to determine also which kind of tube is commonly used: for outpatients, 52 (30%) laboratories use serum or heparinized tubes with separator gel, 46 (24%) of the remaining of laboratories use NaF plasma and heparinized plasma tubes. For hospitalized patients, 99 (52%) laboratories utilizes tubes with separator gel or coagulation activator, while 19 (10%) and 15 (8%) respectively use NaF plasma and heparinized plasma. To perform the Oral Glucose Tolerance Test (OGTT), 84 (43%) laboratories uses tubes without glycolysis inhibitors or separator gel, and only 12 laboratories uses tubes with acidified ternary mixture.
Conclusion: considering these non-completely satisfactory findings, the educational activity of SIBioC working groups on “Diabetes Mellitus” and “Variability of Extra-analytical phase” should be continued, since they appear to be essential for the improvement of the laboratory procedures.

Introduction: the Italian Interdisciplinary Urinalysis Group (GIAU) is a study group aimed to harmonize the standard chemical and morphological urine examination in Italy. The aim of this paper is to present the results of a survey recently released. The survey has been prepared to evaluate the state of the art of the routine standard urine examination in Italy after the issuing by GIAU of the recommendations related to the pre-analytical, analytical and post-analytical phases of the Urine Chemical Morphological Examination (ECMU).
Methods: the questionnaire (47 questions) was made available through the web-based Survey Monkey platform to the members of the two Italian Laboratory Medicine scientific societies (SIBioC and SIPMel).
Results: 553 replies were collected. Regarding the pre-analytical phase, the most critical issue is the use of a monitored (time and temperature) sample transportation to the clinical laboratory only by a small percentage of the laboratories (30%). More than 80% of the laboratories recommends the collection of the mid-stream urine. Regarding the analytical phase, the majority of the participants utilizes dedicated automated analyzers for urine sediment. Only 15.5% of the participants includes the albumin to creatinine ratio in the standard urine examination.
Discussion: the results of the survey give a picture of the state of art of the standard urine examination in Italy. From the comparison with the GIAU previous surveys, it can be concluded that the urinalysis practice in Italy is nowadays more compliant to the indications of the available recommendations and that a general improvement of this laboratory test is observed. Some unsatisfactory answers have been collected for a number of issues, so the role of the GIAU is and will remain fundamental to guide all the laboratory professionals who operate in this laboratory medicine field, towards further advancements.

Introduction: the abuse of New Psychoactive Substances (NPS) has been a major issue since the 1990s, due to the significant impact this phenomenon has on the society and public health. The Italian legislation, in line with the European legislation, is aimed to limit the NPS use in the Country through different intervention strategies.
Methods: the Department for Anti-Drug Policies (DPA) of the Presidency of the Council of Ministers, has activated in Italy the National Early Warning System (SNAP) on NSP reporting, aimed at the early detection of potentially health-threatening phenomena related to the appearance of NPSs in the market. A second aim of the SNAP is the promotion of the technical-analytical expertise of laboratories for the detection of these substances.
Since June 2016, the DPA, where the National Focal Point (NFP) for the verification of the phenomenon of drug dependence is established, has entrusted the coordination of SNAP with the National Centre for Addiction and Doping (NCAD) of the “Istituto Superiore di Sanità”. The Centre works with several partners, including the Pavia Poison Control Centre for clinical toxicology, the Unit of Forensic Toxicology of Sapienza University of Rome for bio-toxicological aspects, and the Central Directorate for Anti-Drug Services for national coordination of seizures operated by law enforcement agencies. To implement this network of collaboration, the NCAD has developed and activated a strategic 'web application' that simplifies the collection of information across the Country, optimizing incoming/outgoing reports between all the involved authorities.
Results: the analysis of the information collected on NPS during 2020 through the SNAP platform, highlighted the circulation of 47 NSPs in Europe, including two synthetic cannabinoids with potential public health risks and the emergence of NPS belonging to the class of benzodiazepines and the benzimidazole family.
Conclusions: The SNAP is an important tool for identifying and notifying the spread of NSPs in our Country very rapidly.

Introduction: The aim of the paper is to present the results from the process of external validation of a number of machine learning (ML) models that had been previously developed to detect SARS-CoV-2 virus positivity on both symptomatic and asymptomatic patients on the basis of the complete blood count (CBC) test.
Methods: Briefly, models were trained using a dataset of 816 COVID-19 positive and 920 negative cases collected at the emergency departments of IRCCS Hospital San Raffaele and IRCCS Istituto Ortopedico Galeazzi. 21 parameters, including the results of the CBC analysis, age [60.9 (0.9) years], gender (57% males) and the presence of COVID-19 related symptoms were used. The validation regarded the evaluation of the error rate (through different metrics, including accuracy, sensitivity, specificity and the area under the curve (AUC)) of the models considered. This external validation was conducted on two well balanced datasets coming from two different hospitals in Northern Italy: Desio hospital and Bergamo Papa Giovanni XXIII hospital. 163 positive and 174 true negative patients from Desio, and 104 positive and 145 true negative from Bergamo were included in the validation.
Results: The performance of the predictive models is satisfactory as we can report an average AUC of 95% for both external datasets.
Conclusion: ML models have been applied to hematological parameters for a more rapid and cost-effective detection of the COVID-19 disease. We make the point that validated models may be useful in the management and early detection of potential COVID-19 patients.

Communication is becoming more important than ever for health care and health care professionals, as the recent COVID-19 pandemic has dramatically highlighted. The fast evolution of the mass and social media and the continuous development of new health-related platforms and applications are imposing new challenges that involve also laboratory medicine and that need to be carefully considered. In fact, these novel, fast and effective strategies of communication are inherently prone to the risk of publishing misleading, incorrect or fake information which can spread uncontrollably and diffuse all over the world in a very short time. However, social media are undoubtedly a great opportunity to communicate, in a responsible and credible way, health-related data and scientific updates and discoveries. As for the therapeutic alliance, it is now required to establish an “information alliance” between different healthcare professionals which, based on a trustworthy relationship, will allow the correct diffusion of health-related information and will contribute to citizens’ education.

Research using biological specimens and health data can accelerate the understanding of many diseases and the effectiveness of preventive, diagnostic and therapeutic interventions. However, researchers have to make a great effort to guarantee respect for transparency and protection of sensitive data of patients, or more generally, of donors whose biological material is collected and stored for future research purposes. Recently, the Committee of Ministers of the Council of Europe proposed new recommendations for the use of biological materials of human origin for future research purposes. The Recommendation CM/Rec(2016)6 focuses on the safeguarding of fundamental rights of the persons, whose biological materials are stored for biomedical research, with particular regard to frail people, unable to provide consent, to guarantee their dignity, integrity and privacy. Furthermore, attention is paid to the transfer of material and data, to the transparency of access policies and to the security in the governance of the collections, in a dimension of globalization and internationalization of research that is a powerful tool to improve human health and healthcare systems.

In recent years, the field of medicine has considerably changed; an important, widespread trend is the reorganization of medical laboratories. These facilities made an effort to improve their efficiency through a process of consolidation and decentralization. In this context, Point-of-Care Testing (POCT) devices offer numerous advantages, but also pose a clinical risk, particularly in Italy, which is limited by its varied, often inadequate, regional regulations. Some examples are reported, representing scenarios of frequent or bothersome POCT use. This document outlines the SIBioC Working Group recommendations for the safe use of POCT instruments. It defines how these have been updated to the state of technologies and medical requirements. Particular attention has been paid to clinical governance, connectivity, the role of the laboratory director and staff, quality control, education, risk management and the role of the in vitro diagnostic companies. Most of the Italian medical practice is based on hospital experience, but the recent pandemic outbreak of SARS-CoV-2 forces us to re-think about the use of POCT in settings that are outside of the hospital such as local clinics, nursing homes and primary care facilities. These facilities will be the next important field of application for those indispensable tools. Networks of instruments, professionals and competence should be carefully tailored to meet local needs and specific clinical settings.

In September 2018 a previously healthy 6-years-old female was admitted for unspecific visual impairment. Physical examination revealed bilateral papilledema with haemorrhage signs, torpid photomotor reflex and slow eye-blink reflex. No other physical abnormalities or alterations in routine laboratory workup were found.
Brain imaging showed alterations in cortical white matter; both optic nerves and cervical spinal cordanti aquaporin-4 antibodies (AQP4-Ab) and myelin oligodendrocyte glycoprotein antibodies (MOG-Ab) testing was then ordered suspecting a neuromyelitis optica spectrum disorder (NMOSD).
MOG-Ab positivity was found at >1:160 and at 1:40 titre in serum and in cerebrospinal fluid respectively. After steroid treatment, the titre in serum was still positive at 1:160, with mild clinical recovery. Clinical and laboratory follow up was maintained for 1 year, highlighting a fluctuating antibody titre related to both the clinical course and the immunosuppressive treatment.
The need of MOG-Ab laboratory follow-up is still debated, nevertheless it might be useful in predicting clinical relapse, as the presentation of this case report illustrates.

Hemoglobin variants are abnormal forms of hemoglobin that may cause hematological disorders. The variants can be (or not) associated with normal hematology parameters. We were able to identify a hemoglobin variant in a 56-year-old Italian Caucasian male using a hematology analyzer.
A routine complete blood count was performed on the Sysmex XN-9000 analyzer for a patient presenting at the Emergency Department with dizziness. Unusual morphological alarms were reported by the analyzer. The microscopy observation of the peripheral blood smear showed a significant number of contracted ed cells and target red cells. Differential count cytograms showed changes in the distribution of the cell clusters and a reduction in the fluorescence signal. In the hypothesis that these alterations were caused by a hemoglobin variant, a HPLC test was performed and a variant in the D/E zone was detected. Molecular analysis confirmed the presence of a variant identified as Hb Leiden. We were thus able to demonstrate the importance to correlate instrumental alarms with validation rules to detect variants in cases where clinical evidence was lacking. The introduction of a new validation rule that automatically activates reticulocyte count and peripheral blood smear in case of specific instrumental alarms is highly advisable.