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

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

Association of Serum Pentraxin-3 and High-Sensitivity C-Reactive Protein with the Extent of Coronary Stenosis in Patients Undergoing Coronary Angiography

Background: We compared factors of inflammation high sensitivity C-reactive protein (hsCRP) and pentraxin-3 (PTX3), and we explored their relationship with coronary artery disease (CAD). Also, we tested the usefulness of hsCRP and PTX3 in the risk assessment of coronary stenosis development and the diagnostic ability of these biomarkers to detect disease severity. Methods: The study group consisted of 93 CAD patients undergoing coronary angiography. Patients were divided into CAD(0), representing subclinical stenosis, and CAD (1-3), representing significant stenosis in one, two or three vessels. Results: We determined the concentration of lipid status parameters, hsCRP and PTX3. We found significantly lower PTX3 and hsCRP concentrations in CAD(0) than in CAD(1-3) group. Concentration of PTX3 showed an increasing trend with the increasing number of vessels affected. The area under ROC curve (AUC) for the combinations of hsCRP and PTX3 with lipid parameters had useful accuracy for detecting CAD(1-3) patients (AUC=0.770, p lt 0.001). Conclusion: PTX3 is a promising independent diagnostic marker for identifying patients with CAD, and a useful indicator of disease progression. In all the analyses PTX3 showed better performance than hsCRP. A combination of PTX3, hsCRP with the lipid status parameters provides risk stratification of the development of coronary stenosis and better classification than their individual application

Circulating resistin protein and mRNA concentrations and clinical severity of coronary artery disease

Introduction: Previous studies have implicated a strong link between circulating plasma resistin and coronary artery disease (CAD). The aim of this study was to evaluate the differences in peripheral blood mononuclear cells (PBMC) resistin mRNA and its plasma protein concentrations between the patients with CAD of different clinical severity. Material and methods: This study included 33 healthy subjects as the control group (CG) and 77 patients requiring coronary angiography. Of the latter 30 was CAD negative whereas 47 were CAD positive [18 with stable angina pectoris (SAP) and 29 with acute coronary syndrome (ACS)]. Circulating resistin was measured by ELISA; PBMC resistin mRNA was determined by real-time PCR. Results: Resistin protein was significantly higher in the ACS group compared to the CG (P = 0.001) and the CAD negative group (P = 0.018). Resistin mRNA expression did not vary across the study groups, despite the positive correlation seen with plasma resistin (rho = 0.305, P = 0.008). In patients, plasma resistin and PBMC resistin mRNA negatively correlated with HDL-C (rho = -0.404, P lt 0.001 and rho = -0.257, P = 0.032, respectively). Furthermore, the highest plasma resistin tertile showed the lowest HDL-C (P = 0.006). Plasma resistin was positively associated with serum creatinine (rho = 0.353, P = 0.002). Conclusion: Significant increase of plasma resistin in patients with ACS compared to CG and CAD negative patients was observed. Despite no change in PBMC resistin mRNA in different disease conditions a positive association between resistin mRNA and resistin plasma protein was evident. Both plasma resistin and PBMC resistin mRNA were negatively associated with plasma HDL-C, and plasma resistin positively with serum creatinine

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

Serumske vrednosti endokana u odnosu na tradicionalne i netradicionalne antropometrijske parametre u populaciji odraslih

Background:Association between endocan and non-traditional anthropometric indices, as distinct cardio -vascular disease risk factors, has not been examined inprevious studies. Endocan is a novel inflammationbiomarker with its higher levels involved in cardiometabolicdiseases development. Taking into consideration thatobesity is an independent risk factor for many cardio -metabolic diseases, we aimed to explore the relationshipbetween endocan levels and novel anthropometric indices i.e., body adiposity index (BAI), cardiometabolic index(CMI), a body shape index, body roundness index, conicityindex, lipid accumulation product index and visceraladiposity index and traditional ones i.e., waist circum -ference, hip circumference, body mass index, waist-to-height ratio and waist-to-hip ratio in adult population.Methods:A total of 177 participants were included.Anthropometric indices and biochemical parametres weremeasured. Results:Univariate regression analysis demonstrated posi-tive correlations of endocan and almost all anthropometricdata. To explore independent associations of endocan andanthropometric parameters, the Model which fulfilled crite-ria for ordinal regression testing was created. Adjusted odds for BAI given in the Model (OR=1.120, 95% CI1.036–1.212, P=0.004), demonstrated that a rise in BAIby 1 unit increased the probability of higher endocan con-centration by 12%. As well, a rise in CMI for 1 unit,increased the probability for higher endocan levels for 2.6times (OR=2.599, 95% CI 1.006–6.712, P=0.049). Atotal of 20.1% of variation in endocan levels could beexplained by this Model.Conclusions:Non-traditional obesity indices, BAI and CMIindependently correlated with higher serum endocan levelsin adult population.Uvod: Povezanost endokana sa netradicionalnim antropometrijskim parametrima, kao različitim faktorima rizika za kardiovaskularne bolesti, nije ispitivana u prethodnim studijama. Endokan je novi biomarker inflamacije, čije su veće vrednosti zabeležene kod kardiometaboličkih poremećaja. Znajući da je gojaznost nezavisan faktor rizika za mnoga kardiometabolička oboljenja, cilj je bio da se ispita povezanost endokana i novih antropometrijskih pokazatelja tj. indeks telesne gojaznosti (BAI), kardiometabolički indeks (CMI), indeks oblika tela, indeks zaokruženosti tela, indeks koniciteta, indeks produkata lipidne akumulacije i indeks visceralne gojaznosti i onih tradicionalnih tj. obim struka, obim kukova, indeks telesne mase, odnos obim struka/telesna visina, odnos obim struka/obim kukova u populaciji odraslih. Metode: Ukupno 177 ispitanika je učestvovalo u istraživanju. Mereni su antropometrijski i biohemijski parametri. Rezultati: Univarijantna regresiona analiza je pokazala pozitivnu korelaciju endokana i skoro svih ispitivanih antropometrijskih parametara. U cilju daljeg ispitivanja postojanja nezavisnih korelacija endokana i antropometrijskih parametara, kreiran je Model koji je zadovoljio kriterijume za ordinalnu regresiju. Prilagođeni Odds-ovi za BAI u Modelu (OR=1,120, 95% CI 1,036-1,212, P=0,004), su pokazali da je porast BAI za 1 jedinicu povećao verovatnoću za porast koncentracije endokana za 12%. Takođe, porast CMI za 1 jedinicu, povećao je verovatnoću za veće vrednosti endokana za 2,6 puta (OR=2,599, 95% CI 1,006-6,712, P=0,049). Ukupno 20,1% varijabiliteta u vrednostima koncentracije endokana može biti objašnjeno ovim Modelom. Zaključak: Netradicionalni pokazatelji gojaznosti, BAI i CMI su nezavisno povezani sa većim vrednostima endokana u populaciji odraslih

Da li je endokan novi potencijalni biomarker za steatozu i fibrozu jetre?

Studies that evaluated endocan levels in nonalcoholic fatty liver disease (NAFLD) and liver fibrosis are scarce. We aimed to explore endocan levels in relation to different stages of liver diseases, such as NAFLD, as determined with fatty liver index (FLI) and liver fibrosis, as assessed with BARD score. A total of 147 participants with FLI≥60 were compared with 64 participants with FLI <30. An FLI score was calculated using waist circumference, body mass index, gamma-glutamyl transferase and triglycerides. Patients with FLI≥60 were further divided into those with no/mild fibrosis (BARD score 0-1 point; n=23) and advanced fibrosis (BARD score 2-4 points; n=124). BARD score was calculated as follows: diabetes mellitus (1 point) + body mass index≥28 kg/m2 (1 point) + aspartate amino transferase/alanine aminotransferase ratio≥0.8 (2 points). Endocan was independent predictor for FLI and BARD score, both in univariate [OR=1.255 (95% CI= 1.104-1.426), P=0.001; OR=1.208 (95% CI=1.029- 1.419), P=0.021, respectively] and multivariate binary logistic regression analysis [OR=1.287 (95% CI=1.055- 1.570), P=0.013; OR=1.226 (95% CI=1.022-1.470), P=0.028, respectively]. Endocan as a single predictor showed poor discriminatory capability for steatosis/fibrosis [AUC=0.648; (95% CI=0.568-0.727), P=0.002; AUC= 0.667 (95% CI=0.555-0.778), P=0.013, respectively], whereas in a Model, endocan showed an excellent clinical accuracy [AUC=0.930; (95% CI=0.886-0.975), P<0.001, AUC=0.840 (95% CI=0.763-0.918), P<0.001, respectively]. Endocan independently correlated with both FLI and BARD score. However, when tested in models (with other biomarkers), endocan showed better discriminatory ability for liver steatosis/fibrosis, instead of its usage as a single biomarkerUvod: Nema mnogo studija koje su ispitivale vrednosti endokana kod obolelih od nealkoholne steatoze i fibroze jetre. Naš cilj je bio da se ispita nivo endokana u različitim stadijumima oboljenja jetre, kao što su nealkoholna steatoza jetre, predstavljena indeksom masne jetre (FLI) i fibroza jetre, predstavljena BARD skorom. Metode: Ukupno 147 učesnika sa FLI≥60 poređeno je sa 64 učesnika sa FLI <30. FLI skor je izračunat koriste i vrednosti obim struka, indeksa telesne mase, aktivnosti gama-glutamil transferaze i vrednosti triglicerida. Ispitanici sa FLI≥60 su dalje podeljeni u 2 grupe: bez fibroze/blaga fibroza (BARD skor 0–1 poen; n=23) i uznapredovala fibroza (BARD skor 2–4 poena; n=124). BARD skor je računat na sledeći način: e erna bolest (1 poen) + indeks telesne mase≥28 kg/m2 (1 poen) + odnos aspartat aminotransferaza/alanin aminotransferaza≥0,8 (2 poena). Rezultati: Endokan je nezavisan prediktor FLI i BARD skora, kako u univarijantnoj [OR=1,255 (95% CI=1,104– 1,426), P=0,001; odnosno OR=1,208 (95% CI=1,029– 1,419), P=0,021], tako i u multivarijantnoj binarnoj logističkoj regresionoj analizi [OR=1.287 (95% CI=1,055– 1,570), P=0,013; odnosno OR=1,226 (95% CI=1,022– 1,470), P=0,028]. Endokan kao samostalan prediktor pokazao je slabu diskriminatornu mo za steatozu/fibrozu jetre [AUC=0,648; (95% CI=0,568–0,727), P=0,002; odnosno AUC=0,667 (95% CI=0,555–0,778), P=0,013], ali je u Modelu pokazao odličnu kliničku tačnost [AUC=0,930; (95% CI=0,886–0,975), P<0,001; odnosno AUC=0,840 (95% CI=0,763–0,918), P<0,001]. Zaključak: Endokan je nezavisno povezan kako sa FLI, tako i sa BARD skorom. Ipak, u modelu (sa drugim biomarkerima), endokan je pokazao bolju diskriminatornu sposobnost za steatozu/fibrozu jetre

Određivanje holesterola u membrani eritrocita

Quantification of cholesterol in biological membranes is an important step toward understanding of metabolism of intracellular cholesterol, composition of cell membrane and plasma lipid profile. The aim of our study was to optimize the method for determination of cholesterol in erythrocyte membrane and then to use this method in the determination of cholesterol concentration in erythrocyte membrane in the studied group of blood samples. Cholesterol in erythrocyte membrane was determined in dry lipid extract of erythrocyte membrane by the enzymatic manual CHOD-PAP method. Cholesterol in erythrocyte membrane and plasma lipid parameters (total cholesterol HDL-cholesterol, LDL-cholesterol and triglycerides) were determined in blood samples of 58 females, obtained by routine health control. Lipid parameters were determined by standard biochemical methods. We examined the relationship between cholesterol in erythrocyte membrane and other plasma lipid parameters as well as other atherogenic risk factors (BMI, blood pressure). Optimization of the method for determination of cholesterol in erythrocyte membrane was based on the observation that primary cholesterol standards prepared by dissolving crystal cholesterol cannot be used due to the interference of the dissolved dry extract in organic solvents with the enzymatic reagent. Commercial standard solutions of cholesterol were used for calibration because they contain detergents for solubilisation of the dry extract in enzymatic reagent. The obtained mean value for cholesterol in erythrocyte membrane, as mmol/L erythrocyte is 4.44 ± 1.019; median 4.65 5-th percentile is 2.70, and 95-th percentile is 6.26. In the examined female group we tested cholesterol concentration in erythrocyte membrane according to age. Two groups were formed (females below and above 50 years) using nonparametric t-test no statistically significant difference was found between these two age groups (p>0.05), while plasma lipid parameters of total cholesterol, triglycerides and LDL-cholesterol were different in the examined groups (p lt 0.05). By Spearmen nonparametric correlation method we found no statistically significant correlation between cholesterol in erythrocyte membrane and other atherogenic risk factors.Određivanje membranskog holesterola je važan stepen u utvrđivanju povezanosti intracelularnog metabolizma holesterola, sastava ćelijske membrane i plazmatskog lipidnog profila. Prvi cilj ovog rada je bio optimizacija metode za određivanje membranskog holesterola što je podrazumevalo izbor standarda i primenu enzimskog testa. Holesterol u membrani eritrocita određivan je ručno u suvom lipidnom ekstraktu membrane, enzimskom CHOD-PAP metodom firme Randox. Optimizirana metoda je primenjena za određivanje koncentracija holesterola u membrani eritrocita kod 58 žena kojima je krv uzeta na rutinskom sistematskom pregledu. U ispitivanim uzorcima određen je i serumski lipidni profil standardnim biohemijskim metodama. Ispitivane žene su podeljene u dve starosne grupe: na mlađe od 50 godina i na starije od 50 godina. Ispitan je i stepen povezanosti između membranskog holesterola sa jedne strane i plazmatskih lipida i drugih faktora rizika za aterosklerozu (starosti, BMI, sistolni i dijastolni pritisak) sa druge strane. Optimizacijom metode za određivanje membranskog holesterola došlo se do zaključka da se ne mogu koristiti primarni standardni rastvori holesterola dobijeni rastvaranjem kristalnog holesterola u organskom rastvaraču zbog slabe rastvorljivosti suvog ekstrakta u enzimskom reagensu i uticaja pojedinih rastvarača na bojenu reakciju. Za kalibraciju treba koristiti komercijalne standardne rastvore holesterola jer sadrže deterdžente koji omogućavaju rastvorljivost suvog ekstrakta. U ispitivanoj populaciji dobijena srednja vrednosti za holesterol u membrani eritrocita (izražene kao mmol/L eritrocita) iznosi 4,44 ± 1,019, mediana 4,65, 5-ti procenat iznosi 2,70, a 95-ti procenat je 6,26. Poređenjem ispitivanih parametara između dve starosne grupe za holesterol u eritrocitnoj membrani nije bilo značajne razlike (p>0,05) dok je za ukupan holesterol, LDL-holesterol i trigliceride ova razlika bila statistički značajna (p lt 0,05). Primenom Spearmanove neparametarske korelacije nije nađena statistički značajna korelacija između membranskog holesterola i drugih faktora rizika za aterosklerozu

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