Association of Adiponectin Receptors with Metabolic and Immune Homeostasis Parameters in Colorectal Cancer: In Silico Analysis and Observational Findings

Adiponectin (ADIPOQ) as both a regulator of metabolic homeostasis and a protein involved in immune response might be of particular interest to contemporary laboratory medicine, especially in terms of minimally invasive diagnostics. The diverse roles of ADIPOQ with regard to the immune and metabolic aspects of colorectal carcinogenesis have been proposed. However, the expression of its receptors ADIPOR1 and ADIPOR2 is scarcely explored in peripheral blood mononuclear cells (PBMCs). Moreover, ADIPORs’ relationships with the immune response mediator TNF-α have not been previously investigated in the PBMCs of CRC patients. This study used both in silico and observational case–control analyses with the aim of exploring the association of ADIPOR gene expression and ADIPOQ single nucleotide polymorphisms (SNPs) with the inflammatory marker TNF-α and lipid status parameters in patients with CRC. Publicly available transcriptomic datasets (GSE47756, GSE44076) obtained from analyses of monocytes and CRC tissue samples were employed for the in silico evaluation of ADIPORs’ specific genetic traits. GSE47756 and GSE44076 datasets were processed with GSEA software to provide a genetic fingertip of different signaling pathways associated with ADIPORs’ mRNA levels. The case–control aspect of the study included the PBMC samples of 73 patients diagnosed with CRC and 80 healthy volunteers. The PCR method was carried out for the PBMC gene expression analysis (ADIPOR1, ADIPOR2, TNF-α mRNA levels) and for the subjects’ genotyping (ADIPOQ rs266729, ADIPOR1 rs7539542). GSEA showed significant associations of ADIPOR mRNA expression with gene sets related to metabolic and immune homeostasis in both datasets. The case–control study revealed the association of ADIPOR1 rs7539542 with reduced lipid status parameters in CRC. In addition, PBMC ADIPOR1 mRNA levels decreased in CRC (p < 0.001), whereas ADIPOR2 mRNA did not differ between the groups (p = 0.442). A reduction in PBMC TNF-α mRNA levels was noted in CRC (p < 0.05). Our results indicate that ADIPOR1 and ADIPOR2 play a significant role in the alteration of both metabolic and immune homeostasis during the progression of CRC. For the first time, ADIPOR1 is shown to be a specific receptor for mediating ADIPOQ’s effects in the PBMCs of CRC patients

Hashimoto Thyroiditis and Dyslipidemia in Childhood: A Review

Hashimoto autoimmune thyroiditis (AIT) is the most common cause of acquired hypothyroidism in the pediatric population. Development of AIT is mediated mainly by cellular immune response directed toward thyroid autoantigens, leading to inflammation and impaired function of thyroid gland. Both thyroid dysfunction and inflammation affect the metabolism of plasma lipoproteins. The alterations in lipid profile worsen with the advancement of hypothyroidism, ranging from discrete changes in euthyroid AIT patients, to atherogenic dyslipidemia in the overt hypothyroidism. In this review, characteristics of dyslipidemia in pediatric AIT patients, and the consequences in respect to the risk for cardiovascular disease (CVD) development are discussed. Additionally, benefit of L-thyroxine treatment on serum lipid profile in pediatric AIT patients is addressed. Finally, potential usefulness of novel lipid biomarkers, such as proprotein convertase subtilisin/kexin type 9 (PCSK9), non-cholesterol sterols, low-density lipoprotein particle size and number, and high-density lipoprotein structure and functionality in AIT patients is also covered. Further longitudinal studies are needed in order to elucidate the long-term cardiovascular outcomes of dyslipidemia in pediatric patients with Hashimoto AIT.articl

Antioksidativna svojstva lipoproteina visoke gustine: više od kardiovaskularne zaštite

High-density lipoprotein (HDL) is the most complex lipoprotein particle, containing lipids and dozens of various functional proteins. Such sophisticated composition enables numerous activities of HDL; from the reverse cholesterol transport, to antioxidative, anti- inflammatory, anti-aggregation, antiadhesive, and vasodilatory effects. Accordingly, the significance of HDL goes far beyond its cardioprotective properties and novel research points towards its role in etiopathogenesis of various other diseases. Antioxidative properties of HDL are primarily attributed to the enzyme paraoxonase 1 (PON1), whose principal role is to protect low-density lipoprotein (LDL) and cell membranes against harmful effects of reactive oxygen species (1). PON1 is located on HDL particles and its activity largely depends on HDL structure. Our investigations have shown that PON1 is not equally distributed across the entire population of serum HDL subfractions. Namely, our results suggest that the allocation of PON1 on specific HDL subclasses changes in pathophysiological conditions, such as chronic kidney disease, polycystic ovary syndrome, or hypertension in pregnancy. Similarly, a shift in HDL subclasses distribution toward smaller, dysfunctional particles is reported in diabetes, metabolic syndrome, obstructive sleep apnea, sarcoidosis, but also in malignant diseases, such as colorectal cancer. In parallel, antioxidative defense mechanisms were diminished in all these categories of patients, which was evident as decreased PON1 activity and rise of oxidative stress. Structural modifications of HDL particles affect their functions, thus antioxidative capability of PON1 depends on qualitative properties of its lipoprotein carrier. Such complex interaction is highly significant for the initiation and progression of numerous diseases.Lipoprotein visoke gustine (high‐density lipoprotein, HDL) je najkompleksnija lipoproteinska čestica koja, pored lipidnih komponenti, sadrži i desetine različitih funkcionalnih proteina. Ovakav složen sastav omogućava brojne funkcije HDL; od reverznog transporta holesterola, do antioksidativnih, antiinflamatornih, antiagregacijskih, antiadhezivnih i vazodilatatornih svojstava. U skladu s tim, značaj HDL u mnogome prevazilazi kardioprotektivne efekte, a savremena otkrića ukazuju na ulogu ovih čestica u etiopatogenezi različitih oboljenja. Antioksidativna svojstva HDL prevashodno su vezana za enzim paraoksonazu 1 (PON1), čija je osnovna funkcija zaštita lipoproteina niske gustine (low‐density lipoprotein, LDL) i ćelijskih membrana od oksidativnih oštećenja (1). PON1 je locirana na HDL česticama i uočeno je da aktivnost ovog enzima u velikoj meri zavisi od strukture samog HDL. U našim istraživanjima pokazali smo da PON1 nije ravnomerno zastupljena na svim subfrakcijama HDL, te da se ova zastupljenost menja u različitim patofiziološkim stanjima, kao što su hronične bubrežne bolesti, sindrom policističnih jajnika, ili hipertenzija u trudnoći. Slično tome, pomeranje raspodele HDL subfrakcija ka manjim, disfunkcionalnim česticama uočeno je kod pedijatrijskih i odraslih pacijenata sa dijabetesom, metaboličkim sindromom, opstruktivnom apnejom u toku spavanja, sarkoidozom, ali i sa malignim bolestima, kao što je kolorektalni karcinom. Istovremeno, nivo antioksidativne zaštite kod ovih pacijenata je bio snižen, što je bilo vidljivo i kao smanjenje aktivnosti PON1, te sledstveno povećanje nivoa oksidativnog stresa. Strukturne modifikacije HDL čestica utiču na njihovu funkciju, pa tako i antioksidativni kapacitet enzima PON1 zavisi od kvalitativnih svojstava njegovog lipoproteinskog nosača. Ovakva složena interakcija između HDL i pridruženih funkcionalnih proteina značajna je za nastanak i progresiju brojnih oboljenja.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Indirect reference intervals for haematological parameters in capillary blood of pre-school children

Introduction: Indirect estimation of reference intervals (RIs) is straightforward and inexpensive procedure for determination of intra-laboratory RIs. We applied the indirect approach to assess RIs for haematological parameters in capillary blood of pre-school children, using results stored in our laboratory database. Materials and methods: We extracted data from laboratory information system, for the results obtained by automatic haematology analyser in capillary blood of 154 boys and 146 girls during pre-school medical examination. Data distribution was tested, and logarithmic transformation was applied if needed. Reference intervals were calculated by the nonparametric percentile method. Results: Reference intervals were calculated for: RBC count (4.2-5.4 x1012/L), haemoglobin (114-146 g/L), MCH (25.0-29.4 pg), MCHC (321-368 g/L), RDW-SD (36.1-43.5 fL), WBC count (4.5-12.3 x109/L), neutrophils count (1.7-6.9 x109/L) and percentage (29.0-69.0%), lymphocytes count (1.6-4.4 x109/L) and percentage (21.9-60.7%), PLT (165-459 x109/L), MPV (8.1-11.4 fL) and PDW (9.2-14.4%). Gender specific RIs were calculated for mo-nocytes count (male (M): 0.2-1.6 x109/L; female (F): 0.1-1.4 x109/L) and percentage (M: 2.5-18.3%; F: 1.8-16.7%), haematocrit (M: 0.34-0.42 L/L; F: 0.34-0.43 L/L), MCV (M: 73.4-84.6 fL; F: 75.5-84.2 fL) and RDW (M: 12.1-14.3%; F: 11.7-13.9%), due to observed gender differences in these parameters (P = 0.031, 0.028, 0.020, 0.012 and 0.001; respectively). Estimated RIs markedly varied from the literature based RIs that are used in the labora-tory. Conclusions: Indirect method employed in this study enables straightforward assessment of RIs in pre-school children. Herein derived RIs differed from the literature-based ones, indicating the need for intra-laboratory determination of RIs for specific populations and sample types

Obstructive sleep apnea and cardiometabolic risk

Opstruktivna apneja u snu (OSA) je hronično progresivno oboljenje sa visokom prevalencom u populaciji koje, bez pravovremene dijagnoze i terapije, može dovesti do značajnih posledica po kvalitet života pacijenata. OSA je čest komorbiditet kod pacijenata sa metaboličkim sindromom (MS) i kardiovaskularnim bolestima (KVB) i predstavlja važan faktor rizika za nastanak ovih oboljenja, a prisustvo nelečenog, teškog oblika OSA povezano je sa porastom ukupnog i mortaliteta usled koronarnih događaja. Brojne studije su ukazale na vezu između MS i OSA, te je ovaj fenomen opisan kao poseban poremećaj - sindrom Z. Istraživanje uzročno-posledične veze između OSA i KVB je u velikoj meri otežano kompleksnom prirodom samog oboljenja. Smatra se da je kardiometabolički rizik u OSA udružen sa arterijskom hipertenzijom, insulinskom rezistencijom, endotelnom disfunkcijom, inflamacijom, dislipidemijom i oksidativnim stresom. Lečenje OSA se danas najefikasnije sprovodi neinvazivnom ventilacijom, pomoću uređaja koji obezbeđuje pozitivan pritisak u gornjim disajnim putevima (eng. continuous positive airway pressure, CPAP) i na taj način sprečava pojavu apneja tokom spavanja. Rezultati kliničkih studija su pokazali da CPAP terapija značajno poboljšava hemodinamske parametre, reguliše hipertenziju, povećava osetljivost na insulin i koriguje dislipidemiju. Buduća istraživanja bi trebalo da rasvetle da li je apneja u snu faktor rizika za KVB per se ili je ta veza posledica šireg patofiziološkog procesa, čiji je deo i OSA.Obstructive sleep apnea (OSA) is a chronic, progressive disorder with a high prevalence in the population. Without timely diagnosis and therapy OSA can significantly affect the quality of life of the patients. OSA is a common co-morbidity in patients with metabolic syndrome (MS) and cardiovascular disease (CVD) and is an important risk factor for their development. The presence of untreated, severe OSA is associated with an increase in total and cardiovascular mortality. Numerous studies have pointed to the relationship between MS and OSA, and this phenomenon was described as syndrome Z. Investigation of the causal relationship between OSA and CVD has been greatly confounded by the complex nature of the disease itself. Cardiometabolic risk in OSA is associated with arterial hypertension, insulin resistance, endothelial dysfunction, inflammation, dyslipidemia, and oxidative stress. The treatment of OSA is now most effectively performed by continuous positive airway pressure (CPAP), a type of non-invasive ventilation which prevents the onset of sleep apnea. The results of clinical studies have shown that CPAP therapy significantly improves haemodynamic parameters, regulates hypertension, increases insulin sensitivity, and corrects dyslipidemia. Future investigations should clarify whether sleep apnea is a risk factor for CVD per se or is a consequence of a broader pathophysiological process, of which OSA is part

Obesity and dyslipidemia

Obesity, a pandemic of the modern world, is intimately associated with dyslipidemia, which is mainly driven by the effects of insulin resistance and pro-inflammatory adipokines. However, recent evidence suggests that obesity-induced dyslipidemia is not a unique pathophysiological entity, but rather has distinct characteristics depending on many individual factors. In line with that, in a subgroup of metabolically healthy obese (MHO) individuals, dyslipidemia is less prominent or even absent. In this review, we will address the main characteristics of dyslipidemia and mechanisms that induce its development in obesity. The fields, which should be further investigated to expand our knowledge on obesity-related dyslipidemia and potentially yield new strategies for prevention and management of cardiometabolic risk, will be highlighted. Also, we will discuss recent findings on novel lipid biomarkers in obesity, in particular proprotein convertase subtilisin/kexin type 9 (PCSK9), as the key molecule that regulates metabolism of low-density lipoproteins (LDL), and sphingosine-1-phosphate (S1P), as one of the most important mediators of high-density lipoprotein (HDL) partides function. Special attention will be given to microRNAs and their potential use as biomarkers of obesity-associated dyslipidemia

Dislipidemija u dijabetes melitusu tip 2

Type 2 diabetes mellitus is a chronic high-prevalence metabolic disease, which is characterized by hyperglycaemia, but also with lipid and protein metabolism disorders. Patients with type 2 diabetes have a high risk for cardiovascular disease (CVD) development and dyslipidemia is considered as a key marker of this increased risk. Hypertriglyceridemia, reduced high density lipoprotein cholesterol (HDL-c) concentrations, and a shift in low-density lipoprotein particles (LDL) distribution toward the small, triglycerides-rich particles, are the most important changes in the lipid profile in diabetes.Type 2 diabetes is a metabolic disorder associated with low grade inflammation and oxidative stress, so in this condition high density lipoprotein particles (HDL) also undergo structural and functional changes and, as a consequence, lose their atheroprotective role. Dyslipidemia treatment in type 2 diabetes patients younger than 40 and without any other risk factor for CVD development starts with changes in a lifestyle, but in patients older than 40 years, first line medications are statins.Glycemic and lipid control in type 2 diabetes patients significantly reduces CVD risk.Dijabetes melitus tip 2 je hronično oboljenje sa visokom prevalencom, koje se karakteriše hiperglikemijom, ali i poremećajima u metabolizmu lipida i proteina. Pacijenti sa tipom 2 dijabetesa imaju visok rizik za razvoj kardiovaskularnih bolesti (KVB) i upravo se dislipidemija smatra ključnim uzročnikom ovog povećanog rizika. Hipertrigliceridemija, snižena koncentracija holesterola u lipoproteinskim česticama visoke gustine (HDL-h) i promena u raspodeli lipoproteinskih čestica niske gustine (LDL) u smeru većeg udela malih čestica, bogatih trigliceridima, predstavljaju najvažnije promene lipidnog profila koje se odnose na dislipidemiju u dijabetesu. Kako je dijabetes stanje kontinuirane blage inflamacije niskog stepena i stalne produkcije slobodnih radikala, HDL lipoproteinske čestice takođe podležu strukturnim i funkcionalnim promenama, usled čega gube svoje ateroprotektivne osobine. Terapija dislipidemije kod pacijenata mlađih od 40 godina bez prisutnih drugih faktora rizika za razvoj KVB počinje promenom životnog stila, a kod pacijenata starijih od 40 godina lekovi izbora u terapiji su statini. Glikemijska i lipidna kontrola kod pacijenata sa tipom 2 dijabetesa značajno umanjuje rizik od nastanka KVB

Cholesterol homeostasis is dysregulated in women with preeclampsia

Introduction The link between preeclampsia and dyslipidemia has been established. Even though lipid profile parameters have been intensively investigated in the pathology of preeclampsia, their accurate molecular mechanisms of action have not been fully decoded. Objectives We aimed to identify the specifics of cholesterol metabolism in women affected by late‑onset preeclampsia and single out potential biomarkers associated with late‑onset syndrome. Patient s and methods A total of 90 pregnant women with a priori risk for preeclampsia were monitored at 4 time points during gestation and, based on the outcome of pregnancy, they were classified into the high‑risk group (70 women) and the preeclampsia group (20 women). Cholesterol metabolic profiling was done using liquid chromatography‑tandem mass spectrometry. Result s The only significant change in the preeclampsia group was an increase in the lathosterol level (P = 0.001). The first‑trimester lathosterol level was higher in the preeclampsia group compared with the high‑risk group (P = 0.02). Further, in the preeclampsia group, positive correlations were found between desmosterol and β‑sitosterol (ρ = 0.474; P = 0.03) in the third trimester, desmosterol and campesterol changes between the second and the first (ρ = 0.546; P = 0.02), and the third and first trimesters (ρ = 0.754; P <0.001), as well as between the desmosterol and β‑sitosterol differences between the third and first trimesters (ρ = 0.568; P = 0.01). No similar correlations were found in the high‑risk group. Conclusions Late‑onset preeclampsia could be associated with an altered lipid profile. By studying the quantitative metabolic signatures of cholesterol, we might assume that both cholesterol synthesis and absorption are increased, that is, there is an imbalance in the cholesterol homeostasis regulation in women affected by the disease

A New Look at Novel Cardiovascular Risk Biomarkers: The Role of Atherogenic Lipoproteins and Innovative Antidiabetic Therapies

The presence of residual cardiovascular disease (CVD) risk is a current dilemma in clinical practice; indeed, despite optimal management and treatment, a considerable proportion of patients still undergo major CV events. Novel lipoprotein biomarkers are suggested as possible targets for improving the outcomes of patients at higher risk for CVD, and their impact on major CV events and mortality have previously been investigated. Innovative antidiabetic therapies have recently shown a significant reduction in atherogenic lipoproteins, beyond their effects on glucose parameters; it has also been suggested that such anti-atherogenic effect may represent a valuable mechanistic explanation for the cardiovascular benefit of, at least, some of the novel antidiabetic agents, such as glucagon-like peptide-1 receptor agonists. This emphasizes the need for further research in the field in order to clearly assess the effects of innovative treatments on different novel biomarkers, including atherogenic lipoproteins, such as small dense low-density lipoprotein (LDL), lipoprotein(a) (Lp(a)) and dysfunctional high-density lipoprotein (HDL). The current article discusses the clinical importance of novel lipid biomarkers for better management of patients in order to overcome residual cardiovascular risk

Izazovi primene hromatografskih tehnika u postavljanju dijagnoze SARS-CoV-2 infekcije

Preventivne mere, rano otkrivanje i potvrda novih slučajeva infekcije, predstavljaju osnov u sprečavanju širenja i suzbijanju ininfektivne bolesti COVID-19. Primena visoko osetljive, spe- cifične real-time RT-PCR metode predstavlja zlatni standard u detekciji SARS-CoV-2. Brza i pouzdana dijanoza su neophodni za efikasno praćenje bolesti, ali veliki broj lažno negativnih slučajeva omogućio je nekontrolisanu transmisiju infekcije. Nove metode za detekciju SARS-CoV-2 iz nazofaringealnog brisa zasnovane na principu tečne hromatografije sa masenom spektrometrijom (HPLC-MS/MS) isključivo se koriste u istraživačke svrhe. Hromatografski testovi omogućavaju istovremenu detekciju više različitih, specifičnih peptidnih markera za identifikaciju SARS-CoV-2. Na ovaj način, moguće mutacije u genskoj sekvenci virusa, lako mogu biti prevaziđene. Upotreba gasne hromatografije sa spektrometrijom pokretljivosti jona (GC-IMS) za detekciju odabranih molekula u izdahnutom vazduhu pacijenata sa COVID-19 može omogućiti neinvazivnu, brzu i tačnu, „point of care” potvrdu dijagnoze bolesti. Uprkos superiornim analitičkim performansama hromatografskih tehnika, njihova primena u rutinskoj laboratorijskoj praksi je retka. Pored opreme, njihova primena zahteva obučeno osoblje i „in house” procedure validacije i verifikacije metoda. Protokoli validacije hromato- grafskih metoda se oslanjaju na preporuke date u naučnim publikacijama i različitim smer- nicama, te su istraživački orijentisani. Najčešće korišćene su EMA, FDA i CLSI smernice za postupke i procedure validacije metoda. Međutim, ove smernice dozvoljavaju različita tuma- čenja i ostavljaju analitičaru da odluči koji od parametara validacije su neophodni. Različiti preanalitički i analitički aspekti hromatografskih metoda diktiraju složenost kriterijuma vali- dacije. Zbog toga je neophodno izdvojiti najvažnije postupke validacije hromatografskih teh- nika (određivanje linearnosti, LOQ i LOD vrednosti, tačnosti i preciznosti metode) i primeniti dostupnu opremu i naučna saznanja