Molecular Laboratory Research Articles

Development and validation of a test system for determining the polymorphism in the DGKH and PPP1R1C genes associated with body weight of sheep

Relevance. National sheep breeding is lagging behind other branches of animal husbandry considering using of modern DNA technologies. The search for new genes — potential candidates associated with economically significant traits — is relevant for a more complete disclosure of the genetic potential of sheep. An earlier genome-wide association study showed that the DGKH and PPP1R1C genes had a certain effect on body weight of sheep at birth and at the age of 90 days. In this regard, a more detailed study of polymorphism in the DGKH and PPP1R1C genes can deepen understanding of growth and development processes in domestic sheep, so we have chosen these genes as targets for the experiment.Methods. Primers and probes were selected for amplification of a fragment with targeted SNPs in the DGKH and PPP1R1C genes with a length of 68 nucleotide pairs based on the reference DNA sequence on the 10th (NC_056063.1) and the 2nd chromosome (NC_056055.1) sheep represented in NCBI. To determine polymorphism, real-time PCR-based test systems were developed. Genotypes were determined using a multiparametric graph. The test systems were tested on 147 sheep of the southern meat breed.Results. The developed test systems for promising DGKH and PPP1R1C genes allowed to clearly determine sheep genotypes based on using the PCR-RT. All three genotypes (T/T, C/T, and C/C) were identified in the DGKH gene. No homozygous genotype (T/T) was identified in the PPP1R1C gene in the studied population. The test systems are suitable for routine DNA analysis in molecular genetic laboratories.

Implementing Next-Generation Sequencing Process Changes to Increase Capacity and Improve Timeliness of Molecular Biomarker Profiling for Lung Cancer Patients.

Faced with expansion of molecular tumor biomarker profiling, the molecular genetics laboratory at Kingston Health Science Centre experienced significant pressures to maintain the provincially mandated 2-week turnaround time (TAT) for lung cancer (LC) patients. We used quality improvement methodology to identify opportunities for improved efficiencies and report the impact of the initiative. We set a target of reducing average TAT from accessioning to clinical molecular lab report for LC patients. Process measures included percentage of cases reaching TAT within target and number of cases. We developed a value stream map and used lean methodology to identify baseline inefficiencies. Plan-Do-Study-Act cycles were implemented to streamline, standardize, and automate laboratory workflows. Statistical process control (SPC) charts assessed for significance by special cause variation. A total of 257 LC cases were included (39 baseline January-May 2021; 218 post-expansion of testing June 2021). The average time for baseline TAT was 12.8 days, peaking at 23.4 days after expansion of testing, and improved to 13.9 days following improvement interventions, demonstrating statistical significance by special cause variation (nonrandom variation) on SPC charts. The implementation of standardized manual and automated laboratory processes improved timeliness of biomarker reporting despite the increasing volume of testing at our center.

Importance of Standard Molecular and Clinical Microbiology Laboratory: Bangladesh Perspective

Abstract Not Available Bangladesh Journal of Medical Microbiology, July 2023;17(2):48-49

Facilitating taxonomy and phylogenetics: An informative and cost-effective protocol integrating long amplicon PCRs and third-generation sequencing

Phylogenetic inference has become a standard technique in integrative taxonomy and systematics, as well as in biogeography and ecology. DNA barcodes are often used for phylogenetic inference, despite being strongly limited due to their low number of informative sites. Also, because current DNA barcodes are based on a fraction of a single, fast-evolving gene, they are highly unsuitable for resolving deeper phylogenetic relationships due to saturation. In recent years, methods that analyse hundreds and thousands of loci at once have improved the resolution of the Tree of Life, but these methods require resources, experience and molecular laboratories that most taxonomists do not have. This paper introduces a PCR-based protocol that produces long amplicons of both slow- and fast-evolving unlinked mitochondrial and nuclear gene regions, which can be sequenced by the affordable and portable ONT MinION platform with low infrastructure or funding requirements. As a proof of concept, we inferred a phylogeny of a sample of 63 spider species from 20 families using our proposed protocol. The results were overall consistent with the results from approaches based on hundreds and thousands of loci, while requiring just a fraction of the cost and labour of such approaches, making our protocol accessible to taxonomists worldwide.

A study of the Relationship of Polymorphisms of the PPARGC1A Gene with Milk Production and its Components, in Holstein Friesian Cows

This research was done at a Molecular Genetics lab at the College of Agriculture at the University of Basrah. The objective of this study is to determine how Holstein Friesian cows’ PPARGC1A genetic polymorphism affects milk production and its constituent parts. For milk production and quality standards, 59 Holstein Friesian cows from Taj Al-Nahrain station in Al-Diwaniyah are employed. Blood was drawn for genomic DNA, and the PPARGC1A gene was amplified using PCR. The samples were then transferred to the Korean firm Macrogene for DNA sequencing analysis. Morning milk 50ml samples were collected once each week., and after taking the samples during the milking process, they were evaluated for quality using the Julie Z7 milk analyzer. Association analysis was performed using SPSS statistical software. According to the current study’s findings, the stage of milk production had a significant impact on both daily and total production (p ≤0.05). The results also shows that there is a significant differences (p≤0.05) of stage of milk production on the chemical composition of milk, while there are no significant differences between Genetic Polymorphisms in daily and total milk production. The results show that hap1 is superior on hap2 in the chemical contents of milk, except for the percentage of milk fat, which has not reach a significant level.

COVID-19 Pandemic: Total Testing Cost Impact on Neisseria Gonorrhoeae and Chlamydia Trachomatis Testing and Positivity Rate in the Veteran Population

Abstract Introduction/Objective The College of American Pathologists (CAP) checklist includes the quality assurance (QA) monitoring of the rate of diagnosis of Neisseria Gonorrhoeae (NG) and/or Chlamydia trachomatis (CT) by the molecular laboratory. Recently, the societal impacts of the economic lockdowns of the COVID-19 pandemic had a significant effect on the number of tests. However, the impact on the total testing costs as well as the positivity rate has not been well studied in the Veteran population. Methods/Case Report Quarterly quality assurance data from October 1st, 2018, to December 31st, 2022, was reviewed to obtain both the test volume for CT/NG and the number of positive test results at a Veterans Affairs Medical Center (VAMC). In addition, the cost to perform each test in reagents and consumables was also noted. Quarter 1 at the VAMC begins in October of the prior calendar year. The quarters in the period of April 1st, 2020, to December 31st, 2020 (corresponding to Quarter 3 2020 to Quarter 1 2021), was analyzed and compared to both the quarterly data prior to and after this period to note the impact of the COVID-19 pandemic on the quality assurance data. Testing had been performed using the Abbott m2000 RealTime System (Abbott Park, Illinois). Results (if a Case Study enter NA) A total of 18,904 tests for CT/NG had been performed on the Veteran population during the study period. Of these 18,904 tests, 385 were positive for NG and 515 were positive for CT. Quarterly percentage rates on a quarterly basis ranged from 1.58% to 4.53% (CT) and 1.16% to 4.53% (NG) with average rates of 2.78% and 2.13%, for Chlamydia and Gonorrhea respectively. The total reagent and consumable cost for each test was $28.84 per test. The period of Quarter 3 2020 to Quarter 1 2021 was marked by a decrease in total tests (331, 713 and 781 tests respectively compared with 1,049 tests in the quarter just before this period) with a corresponding decreased total test cost expenditure ($9,546.04, $20,592,92, and $22,524.04 respectively compared with $30,253.16 in the quarter just before this period) for performed tests. There was also an increase in the positivity rate (the highest positivity rate for both CT and NG of 4.53% was noted in quarter 3 2020) in this period of Quarter 3 2020 to Quarter 1 2021. Conclusion The initial period of the COVID-19 pandemic was marred by widespread economic lockdowns that impacted the total number of CT and NG tests performed, total test expenditures for tests performed, as well as the positivity rate.

Appetite- and Weight-Regulating Neuroendocrine Circuitry in Hypothalamic Obesity.

Since hypothalamic obesity (HyOb) was first described over 120 years ago by Joseph Babinski and Alfred Fröhlich, advances in molecular genetic laboratory techniques have allowed us to elucidate various components of the intricate neurocircuitry governing appetite and weight regulation connecting the hypothalamus, pituitary gland, brainstem, adipose tissue, pancreas, and gastrointestinal tract. On a background of an increasing prevalence of population-level common obesity, the number of survivors of congenital (eg, septo-optic dysplasia, Prader-Willi syndrome) and acquired (eg, central nervous system tumors) hypothalamic disorders is increasing, thanks to earlier diagnosis and management as well as better oncological therapies. Although to date the discovery of several appetite-regulating peptides has led to the development of a range of targeted molecular therapies for monogenic obesity syndromes, outside of these disorders these discoveries have not translated into the development of efficacious treatments for other forms of HyOb. This review aims to summarize our current understanding of the neuroendocrine physiology of appetite and weight regulation, and explore our current understanding of the pathophysiology of HyOb.

Simplifying molecular testing through electronic order entry

Abstract Our solid-tumor molecular laboratory receives more than 3,000 orders each year, with about two-thirds coming from internal providers. Before this project, molecular tests were ordered using one of two different paper forms, which the care team then faxed or e-mailed to the Genetics Preanalytical Service (GPS). These requests led to frequent modifications and cancellations due to the complexity of molecular testing. A small fraction of requisitions were lost due to transmission errors. We undertook a project to design, develop, and implement computerized physician order entry (CPOE) in Epic (Epic Systems Corp., Verona, WI) and Sunquest (Clinisys, Tucson, AZ). We conducted extensive interviews with the directors of the molecular lab, laboratory medicine IT, GPS, anatomic pathology staff, laboratory scientists, laboratory genetic counselors, oncologists, clinicians, and nurse coordinators. The order itself and other technical changes underwent iterative review and testing by a subset of interviewees. Five oncologists participated in formal beta-testing. The user interviews allowed us to produce a detailed process diagram that drew attention to specific areas of repetition or risk. We proposed a new workflow that would eliminate repetition, provide redundant order-tracking, and automate some lab processes. Once approved, we built the electronic order in Epic and modified existing laboratory programs and interfaces as needed. The process of placing an order, which previously typically involved both nurses and doctors over several calendar days, was now being completed in fewer than 5 minutes by a single practitioner. Requisition status events were no longer created manually in Sunquest by GPS. Certain data-entry tasks were eliminated through the incorporation of label printers. Within one month of launch, more than 50% of somatic molecular orders arrived via CPOE, increasing to 90% within six months. At a rate of 125 CPOE orders per month, time savings range from 20 hours per month for GPS staff, 50 hours per month for nursing, and 20 hours per month for physicians. The success of this project demonstrates the feasibility of replacing a paper-based process for a complex test menu with CPOE. Due to the intricacies of molecular testing, such a change will undoubtedly involve multiple divisions. We attribute the success of this project to early involvement by key stakeholder groups, including developers; an extended testing phase to ensure reliability at roll-out; creation of straightforward reference documents; and rapid response to user questions, especially during the first weeks post-launch.

Prevalence of Covid-19 and Associated Co-Morbidities at a Tertiary Hospital in Eastern Nepal

Introduction: COVID-19 (coronavirus disease) manifests as asymptomatic, mild, or severe pneumonia-like symptoms. COVID-19 patients with diabetes, chronic obstructive pulmonary disease (COPD), cardiovascular disease (CVD), hypertension, malignancies, HIV, and other comorbidities could develop a life-threatening condition.
 Objectives: To identify the various comorbid illnesses in COVID-19-positive patients along with its baseline characteristics in a tertiary hospital in eastern Nepal.
 Methodology: A one-month observational study carried out in RT-PCR positive COVID-19 patients in Birat Medical College and Hospital (BMCTH) Molecular Laboratory from June to July 2021. All the data was entered in a pre-structured proforma. The data was then further entered and analyzed by using Microsoft Excel 2014.
 Result: There were 354 (22.77%) RT-PCR positive COVID-19 cases and 1200 (77.22%) RT-PCR negative COVID-19 cases. More than half of the COVID-19 positive patients were males 211(59.60%). The most common symptoms were fever (92.3%), cough (83.5%) and tiredness (81.23%). Psycho-social stress such as anxiety, depression and insomnia 123 (34.74%) were common comorbidities followed by diabetes 91(25.70%) and hypertension 72(20.33%).
 Conclusion: COVID-19 patients were mostly between the 20-30 years of age group. Fever, cough, and throat pain were the commonest presenting symptoms.

Yeast recombinational cloning for heterologous biosynthesis of polyketides: a molecular microbiology laboratory module for undergraduate students.

Recombinant plasmids are essential tools in molecular biotechnology, and reliable plasmid assembly methods have, therefore, become a prerequisite for the successful cloning and transfer of genes. Among the multitude of available plasmid assembly strategies, in vivo homologous recombinational cloning in yeast has emerged as a cost-effective and relatively simple method. Since we use this method routinely in our group for assembling large plasmids with secondary metabolite gene clusters and for direct heterologous production of polyketides in Saccharomyces cerevisiae, we developed an exercise module for undergraduate students where they would get hands-on experience with these molecular practices. The exercises target several molecular techniques, including PCR, restriction enzyme digestion, and yeast recombinational cloning. The students will learn about plasmid assembly and yeast transformation methods by performing these experiments while inherently acquiring new skills valuable for their subsequent laboratory work or projects.

Babesia negevi infection in dogs and response to treatment

Canine babesiosis is an important protozoan tick-borne disease associated with anemia and thrombocytopenia and caused by several different Babesia spp. Babesia negevi was first reported to infect dogs in the Middle East in 2020. This study describes the presentation, clinical signs, parasitemia levels quantified by molecular techniques, laboratory findings and treatment of dogs infected with B. negevi following the first description of this species. Clinical findings in the infected dogs, a 3-year old female and two 8-week old male and female pups, included extreme lethargy and pale mucous membranes, anemia and thrombocytopenia found in all three animals. Fever was present in the older female and icterus in the female pup. Babesia parasites resembling B. negevi were detected by microscopy of blood smears from the dogs. PCR of blood targeting the 18S rRNA and cox1 genes confirmed that babesiosis was caused by B. negevi and PCR targeting the Borrelia flagellin gene indicated co-infection with Borrelia persica in two dogs. Treatment of the dogs with imidocarb dipropionate resulted in clinical improvement and initial decrease in the B. negevi parasite load as detected by quantitative PCR in two dogs, however the female pup continued to deteriorate and died. The parasite load in the 3-year old female decreased from 43,451 parasites/µl blood pre-imidocarb dipropionate treatment to 803 parasites/µl within two weeks. In the surviving pup, it decreased from 3,293,538 parasites/µl pre-treatment to 20,092 parasites/µl after two weeks. Babesia negevi DNA was still recovered from blood samples by PCR despite repeated treatment with imidocarb dipropionate one-month post-treatment in the surviving pup and up to seven months post-treatment in the 3-year old female. Only treatment with atovaquone and azithromycin for ten days eliminated B. negevi in both dogs as confirmed by negative PCR two weeks later. In conclusion, treatment with imidocarb dipropionate was helpful for recovery from clinical disease but did not facilitate parasite elimination, and it is therefore recommended to treat canine B. negevi infection with the combination of atovaquone and azithromycin.

A Maltese landscape of cardiac genetics

Abstract Introduction Advances in human genetics have improved our understanding of various inherited cardiovascular conditions (ICC). Genetic testing is now an important pillar in the management of patients suffering from cardiomyopathies, channelopathies and aortic and other vascular disorders. Existing population databases may not cater for the local gene pool. The local genetic yield of ICCs has never been explored. Objective The aim of this study was to evaluate the population referred for genetic testing and to assess the genetic yield according to the referring phenotype. Methodology Probands referred for genetic testing between 2018-2022 were included. Data was retrospectively reviewed in close liaison with the molecular genetics lab at a university. Variants classified as pathogenic or likely pathogenic by ACMG criteria were labelled as ‘gene positive’ for the purpose of this study. Results 328 patients were referred for genetic testing (63.7% Male, 51.3±18.7 years). The majority were referred because of a cardiomyopathy phenotype (64.02%) followed by arrhythmic disorders (20.7%), aortic/vascular disorders (12.5%), familial hyperlipidaemia (1.5%) and complex congenital heart disease (0.6%). Two (0.6%) did not have a reason specified. 94 (28.7%) probands were gene positive, 5 patients (1.5%) had two mutations, 41.4% of all mutations were novel. The highest genetic yield was seen in cardiomyopathy patients (34.8%), followed by aortic/vascular disorders (24.4%) and arrhythmic disorders (14.7%). A separate analysis was also performed for each individual phenotype (Table 1). The dominant genes for each condition: MYH7 (24.4%) and MYBPC3 (22.0%) for hypertrophic cardiomyopathy; TTN (52.9%) and FLNC (23.5%) for dilated cardiomyopathy; DSG2 (45.5%) and FLNC (18.2%) for arrhythmogenic cardiomyopathy; TTN (40.0%) and MYH7 (40.0%) for left ventricular non-compaction cardiomyopathy; TTR (100%) for cardiac amyloid; SCN5A (37.5%) for non-specific cardiomyopathy; KCNH2 (50.0%) and KCNQ1 (50.0%) for long QT syndrome; SCN5A (37.5%) for Brugada Syndrome; FBN1 (40.0%) for skeletal phenotype; FBN1 (33.3%) for aortic phenotype; and LDL-R (100.0%) for familial hyperlipidaemia Conclusion The overall ICC genetic yield in Malta is 28.7%. A substantial proportion were novel mutations. The highest genetic yield is in the cardiomyopathy group. The yield for most phenotypes is comparable to other labs. The proportion of gene positive LQTS patients is surprisingly very small, a phenotype with an established high genetic yield in various studies (up to 80%). Clinicians caring for patients with ICCs now have local diagnostic yield data to better counsel patients when referred for genetic testing.

Clinical implications of PNA‑sequencing as a complementary test for EGFR mutation analysis in human lung cancer.

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are the first-line regimen for the treatment of non-small cell lung cancer (NSCLC) patients with EGFR mutations. However, false-negative results are occasionally observed, even with FDA-approved molecular tests. Such examples in have been reported in our pilot study showing a slightly upward-shifted amplification curve using commercial reverse transcription-quantitative (RT-q)PCR. Verification using peptide nucleic acid (PNA) clamping-sequencing, which has a sensitivity of ~0.1%, may allow better prediction of which patients will benefit from EGFR-TKI therapy. To confirm this hypothesis, samples were prospectively collected from 1,783 lung cancer cases diagnosed in National Cheng Kung University Hospital between 2012-2018. An independent lung cancer cohort of 1,944 cases was also recruited from other hospitals. The clinical significance of mutant-enriched PCR with PNA-sequencing was analyzed and patient outcomes were followed. A total of 17 of 34 cases (50%) were found to harbor EGFR mutations by PNA-sequencing. A total of 22 cases were discovered in the independent lung cancer cohort, and 14 of these (63.6%) cases had EGFR mutations. TKIs were administered to 14 of the 17 mutation-positive patients, and a partial response was observed in 4 cases and stable disease in 10 cases. Patients with EGFR mutations receiving a TKI regimen had a longer overall survival (OS) (median: 40.0 vs. 10.0 months) compared with those without treatment. The difference in OS was not significant. Based on the results of the present study, combining RT-qPCR with PNA-sequencing may be a practical supplementary technology in a clinical molecular laboratory for a subset of lung cancer patients in selection of EGFR TKI therapy.

PIK3CA testing in HR+/HER2− Metastatic Breast Cancer: Real-world data from referral molecular pathology labs

PIK3CA testing in HR+/HER2− Metastatic Breast Cancer: Real-world data from referral molecular pathology labs

A Novel Approach on the Sensitivity of Preserved Plasmodium falciparum Positive RDT kits for the Molecular Detection of Pfmdr 1 and Pfcrt Genes in Pregnant Women

Storing blood samples for molecular screening have become increasingly difficult due to unstable electricity and high cost of generators fuels. Samples often deteriorate giving false negative upon molecular analysis. This research was aimed at determining the sensitivity of preserved Plasmodium falciparum positive RDT kits for the molecular detection of Pfmdr 1 and Pfcrt genes among pregnant women in From February 2020 to May 2020, Sokoto, Nigeria. 200 pregnant patients who visited the hospital for ANC had their blood samples taken, and a questionnaire was given out. To detect Plasmodium falciparum, RDT kits tailored to its monoclonal antibody were utilized. P. falciparum positive Rapid Diagnostic Test Kits were air tightened with nylon and silica-preserved for three months in a closed cabinet. The preserved positive samples were utilized to extract DNA, and the Pfmdr1 and Pfcrt genes' existence was then checked using a PCR procedure. 4 samples tested positive for both genes, suggesting a 21.05 % prevalence, according to the results of the inquiry into the Pfcrt and Pfmdr1 genes. This study has revealed the sensitivity of preserved RDT kits (with silica) for DNA extraction. Researchers and molecular laboratories should be preserving positive RDT kits meant for DNA extraction and other molecular analysis as it is much easier to preserve and there is no cost involved.

Analysis of the viscosity and self-healing performance of warm-mix recycled asphalt: A method combining molecular dynamics simulations and laboratory tests

The technology of warm-mix regeneration has been frequently used in the production of asphalt pavement; however, the viscosity and self-healing properties of warm-mix recycled asphalt (WMRA) need further study. Therefore, the viscosity and self-healing properties of WMRA were analyzed by combining laboratory tests and molecular dynamics (MD) simulations. First, WMRA with different SWA and reclaimed asphalt pavement (RAP) contents were prepared in the laboratory. Then, different simulations, including viscosity simulation and self-healing simulation and Brookfield Viscosity test, were developed to analyze the performance variation rule. Experimental results showed that the Sasobit warm-mix agent (SWA) significantly reduced asphalt viscosity, and the recycled asphalt’s viscosity increased with asphalt aging deeply. In contrast, the addition of 2% SWA made the viscosity of 100% RAP similar to that of new asphalt. Considering the viscosity reduction effect, asphalt performance and economy, a 3% SWA content is recommended. Both SWA and the degree of aging can reduce the healing performance of WMRA. There is little difference between the two factors in reducing self-healing performance. In general, the study's findings serve as a useful guide for the performance analysis of WMRA.

Comparative diagnostic performance of a Cas13-based assay for detecting COVID-19 cases in Al-Dewaniyah province, Iraq.

Effective, accurate, and rapid diagnostic tools for detecting SARS-Covid2 infection are urgently needed to prevent and control the pandemic. While RT-qPCR is a gold standard test for diagnosing SARS-Covid2 infections, this method is limited by the requirement for expensive equipment, experienced staff and specialized molecular laboratory. Recently, the CRISPR cas13 platform was used for the detection of COVID-19. This study evaluated the clinical performance of the CRISPRbased cas13a diagnostic assay for SARS-Covid2. In the current study, the diagnostic performance of the Cas13-based SHERLOCK (Specific High Sensitivity Enzymatic Reporter Unlocking) assay was compared with RT-qPCR in terms of sensitivity, specificity and time consumption. Keywords: Crispr-Cas13; Covid-19; Sherlock

Hemp Abnormal Growth Is Attributed to Mono-, Di-, or Tri-Infections of Spiroplasma citri, ‘Candidatus Phytoplasma trifolii’, and Beet Curly Top Virus

As hemp ( Cannabis sativa) emerges as a commercial crop in Nevada, mild to severe abnormal growth has been observed from many plants in commercial fields throughout the growing season. Affected plants exhibited a group of symptoms typically including stunting, leaf yellowing, excessive apical branching, clusters of witches’ broom, leaf rolling upwards, and leaf mottling and mosaic. These symptoms mostly showed up together in a plant or a crop and are defined here as abnormal growth syndrome (AGS). Between 2017 and 2019, the Nevada State Plant Pathology and Molecular Diagnostic Lab received 67 symptomatic hemp samples from Nevada for diagnosis, and ‘ Candidatus Phytoplasma trifolii’ was detected in 14 samples (21%). To investigate additional biotic agents associated with AGS, PCR products generated by primers P1/P7 were cloned into a pGEM-T vector and sequenced, and Spiroplasma citri DNA was found in two samples (3%). All 67 DNA samples were further tested for beet curly top virus (BCTV), and 57 samples (85%) were found to be infected by BCTV. Twelve samples (18%) were coinfected by both ‘ Ca. Phytoplasma trifolii’ and BCTV, and two (3%) were coinfected by S. citri, ‘ Ca. Phytoplasma trifolii’, and BCTV. The findings suggest that BCTV is the most prevalent pathogen causing the hemp abnormal growth in Nevada, but ‘ Ca. Phytoplasma trifolii’ and S. citri may also contribute to the severity and complexity of symptoms. Thus, hemp abnormal growth can be attributed to single, dual, or triple infections of these three leafhopper-vectored mollicutes and virus. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Platelets as Potential Non-Traditional Cardiovascular Risk Factor-Analysis Performed in Healthy Donors.

Abnormal lipid profile, increased glucose level, and elevated body weight are traditional cardiometabolic risk factors; however, the role of platelets in the development of cardiovascular disease (CVD) is increasingly being highlighted. The aim of this study was to select platelet-related parameters (non-genetic molecular and routine laboratory measurements) that may be associated with increased cardiovascular risk among healthy populations. We evaluated the level of platelet indices, platelet-based inflammatory markers, platelet reactivity parameters, and platelet reactive oxygen species (ROS) generation in relation to selected cardiometabolic risk factors. We noted the association between total cholesterol and LDL cholesterol with platelet aggregation and platelet ROS generation. We found the relationship between triglycerides, glucose, and body mass index with the relatively new multi-inflammatory indices (MII-1 and MII-3). Moreover, we noticed that the mean platelet volume-to-lymphocyte ratio in healthy subjects is not a good source of information about platelets and inflammation. We also highlighted that platelet-to-HDL-cholesterol ratio may be a promising prognostic cardiometabolic indicator. The association between platelet-related (especially molecular) and cardiometabolic parameters requires further research. However, the goal of this study was to shed light on the consideration of platelets as a non-traditional cardiovascular risk factor and a crucial element in identifying individuals at high-risk of developing CVD in the future.

Platelet distribution width is associated with cardiovascular mortality in an adult general population

Platelet distribution width (PDW), a marker of platelet size heterogeneity used as a readout of processes leading to platelet production and destruction, was recently reported to tag platelet activation variability. As platelets participate in the pathogenesis of many acute and chronic diseases, we evaluated PDW as a predictor of all-cause and cause-specific mortality. Longitudinal analysis was performed on 17,334 participants (52% women, mean age 55.6±12 years) in the Moli-sani study cohort, without a history of hematological diseases. Baseline PDW measurements were categorized in tertiles, the lowest acting as the reference. A multivariable Cox-proportional hazard model was used to estimate the association between PDW and mortality. Over a median follow-up of 11.6 years (interquartile range 10.7-12.5), 1,535 deaths [37.7% cardiovascular disease (CVD) and 36.5% cancer] were ascertained. As compared to those in the first PDW tertile (14.6-16.0 fL), individuals within the highest tertile (16.6-20.4 fL) had an increased risk of all-cause [hazard ratios (HR):1.20; 95% CI: 1.04-1.37] and CVD mortality (HR:1.29; 1.03-1.62). No association between PDW and cancer mortality was found in the whole sample. Subgroup analyses by two age classes (35-65y, ≥65y) showed that the association of PDW with both all-cause and cancer mortality was more apparent in the elderly (HR:1.34; 1.14-1.58, P for interaction =0.028 and HR:1.37; 1.01-1.85, P for interaction =0.020, respectively). We conclude that PDW-associated increase in CVD mortality risk could be related to accelerated/altered activation, production, or destruction of platelets, leading to several clinical conditions and death. In the elderly, PDW involvement in all-cause and cancer mortality should be further investigated. *Moli-sani investigatorsSteering committee: Licia Iacoviello, Giovanni de Gaetano, Maria Benedetta Donati. Scientific secretariat: Marialaura Bonaccio, Americo Bonanni, Chiara Cerletti, Simona Costanzo, Amalia De Curtis, Augusto Di Castelnuovo, Alessandro Gialluisi, Francesco Gianfagna, Mariarosaria Persichillo, Teresa Di Prospero. Safety and ethical committee: Jos Vermylen, Renzo Pegoraro, Antonio Spagnolo. External event adjudicating committee: Deodato Assanelli, Livia Rago. Baseline and follow-up data management: Simona Costanzo, Marco Olivieri, Teresa Panzera. Data analysis: Augusto Di Castelnuovo, Marialaura Bonaccio, Simona Costanzo, Simona Esposito, Alessandro Gialluisi, Francesco Gianfagna, Sabatino Orlandi, Emilia Ruggiero, Alfonsina Tirozzi. Biobank, molecular and genetic laboratory: Amalia De Curtis, Sara Magnacca, Fabrizia Noro, Alfonsina Tirozzi. Recruitment staff: Mariarosaria Persichillo, Francesca Bracone, Teresa Panzera. Communication and press office: Americo Bonanni. Regional institutions: Direzione Generale per la Salute - Regione Molise; Azienda Sanitaria Regionale del Molise; Agenzia Regionale per la Protezione Ambientale del Molise; Molise Dati Spa; Offices of vital statistics of the Molise region. Hospitals: Presidi Ospedalieri ASReM: Ospedale A. Cardarelli, Campobasso; Ospedale F. Veneziale, Isernia; Ospedale San Timoteo, Termoli (CB); Ospedale Ss. Rosario, Venafro (IS); Ospedale Vietri, Larino (CB); Ospedale San Francesco Caracciolo, Agnone (IS); Casa di Cura Villa Maria, Campobasso; Ospedale Gemelli Molise, Campobasso; IRCCS Neuromed, Pozzilli (IS), Italy.