Effect of estradiol on regulation of endothelial and inducible nitric oxide synthase in the heart of obese rats

Estradiol ostvaruje pozitivan efekat na kardiovaskularni sistem (KVS), sprečavanjem nastanka ateroskleroze i endotelne i/ili vaskularne disfunkcije, a deluje i direktno na srce smanjujući njegovu hipertrofiju. Estradiol ostvaruje pozitivne efekte u KVS regulacijom azot-monoksid (NO)-sintaza (NOS), endotelne (eNOS) i inducibilne (iNOS) forme, koje su odgovorne za sintezu NO, aktivacijom sloţene mreţe unutarćelijskih signalnih puteva, u koje su uključeni signalni molekuli supstrat receptora za insulin 1 (IRS-1), fosfatidil-inozitol-3-kinaza (PI3K) i protein kinaza B (Akt). Estradiol utiče na signalnu transdukciju angiotenzina II (Ang II), delujući na ekspresiju receptora za Ang II tipa 1 (AT1R) i tipa 2 (AT2R), kao i na nivo RhoA proteina. Estradiol utiče i na transport energetskih supstrata regulišući translokazu masnih kiselina (CD36) i transportere za glukozu tipa 1 (GLUT1) i tipa 4 (GLUT4). Kontrolni i gojazni muţjaci pacova soja Wistar tretirani su estradiolom (40μg/kg) ili 1% etanolom 24 sata pre ţrtvovanja. Spektrofotometrijskim metodama određivane su koncentracije NO u plazmi i koncentracije L-Arginina (L-Arg), NO i slobodnih masnih kiselina (SMK) u lizatu srca. Western blot metodom određivani su nivoi proteina eNOS, iNOS, NF-κB-p65, pIRS-1/IRS-1, p85 i p110 subjedinice PI3K, pAkt/Akt, RhoA, AT1R, AT2R, GLUT1, GLUT4 i CD36 proteina, kao i asocijacija IRS-1 sa p85-PI3K proteinom u srcu pacova, dok je imunohistohemijskom metodom određivana ekspresija i lokalizacija eNOS i iNOS. Metodom qRT-PCR određivan je nivo iRNK za eNOS i iNOS u srcu pacova. Rezultati su analizirani korišćenjem Studentovog t-testa. Rezultati prikazani u ovoj doktorskoj disertaciji pokazuju da tretman estradiolom kod kontrolnih pacova povećava nivo p85 subjedinice PI3K i fosforilaciju Akt na Thr308, dok smanjuje ekspresiju gena za iNOS...Estradiol has a positive effect on the cardiovascular system (CVS) by preventing the development of atherosclerosis and endothelial and/or vascular dysfunction. Estradiol exerts many of its effects on the CVS by regulation of nitric oxide synthase (NOS), endothelial (eNOS) and inducible (iNOS) forms that are responsible for the synthesis of nitric oxide (NO), through activation of the complex network of intercellular signal pathways that include signaling molecules: insulin receptor substrate 1 (IRS-1), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt.) Estradiol also affects the signal transduction of angiotensin II (Ang II) by affecting the expression of Ang II receptors: type 1 (AT1R) and type 2 (AT2R), as well as the level of RhoA protein. Finally, estradiol indirectly regulates the transport of energy substrates in heart, by regulating transporters of free fatty acids (FFA) and glucose, CD36 and glucose transporters type 1 (GLUT1) and type 4 (GLUT4). Control and obese male Wistar rats were treated with one dose of estradiol (40 μg/kg) or with the same amount of 1% ethanol in saline 24 hours before sacrificed. The concentration of NO was measured in the plasma and in lysates, while the concentrations of L-Arginine and FFA were determined in the lysates of the rats‘ hearts. The Western blot method was used to determine the level of eNOS, iNOS, NF-kB p65, IRS-1, p85 and p110 subunits of PI3K, Akt, AT1R, AT2R, RhoA, CD36, GLUT1 and GLUT4 in the rats‘ heart, as well as association of IRS-1 with p85 subunit of PI3K. Immunohistochemical analysis was used for localization and expression of eNOS and iNOS proteins in heart. qRT-PCR was used to determen the level of iRNA of eNOS and iNOS in rats‘ heart. Results were analyzed using Student‘s t test..

Insulin-like growth factor-1 reduces cardiac autosis through decreasing AMPK/FOXO1 signaling and Na+/K+-ATPase-Beclin-1 interaction

Introduction: Insulin-like growth factor-1 (IGF-1) promotes survival and inhibits cardiac autophagy disruption. Methods: Male Wistar rats were treated with IGF-1 (50 μg/kg), and 24 h after injection hearts were excised. The level of interaction between Beclin-1 and the α1 subunit of sodium/potassium-adenosine triphosphates (Na+/K+-ATPase), and phosphorylated forms of IGF-1 receptor/insulin receptor (IGF-1R/IR), forkhead box protein O1 (FOXO1) and AMP-activated protein kinase (AMPK) were measured. Results: The results indicate that IGF-1 decreased Beclin-1’s association with Na+/K+-ATPase (p < 0.05), increased IGF-1R/IR and FOXO1 phosphorylation (p < 0.05), and decreased AMPK phosphorylation (p < 0.01) in rats’ hearts. Conclusions: The new IGF-1 therapy may control autosis and minimize cardiomyocyte mortality

Subacute thyroiditis following COVID-19 vaccination: Case presentation

Background:Subacute thyroiditis (SAT) is an organ-specific disease that various drugs, including COVID-19vaccines, can trigger. COVID-19 infection has been associated with thyroid gland damage and disease SARS-CoV-2direct action, euthyroid sick syndrome, and immune-mediated mechanisms are all potential mechanisms of thyroiddamage. It denotes thyroid gland inflammation, most commonly of viral origin, and belongs to the transitory, self-limiting thyroid gland diseases group, causing complications in approximately 15% of patients in the formof permanent hypothyroidism. Some authors say SAT is the most common thyroid disease associated withCOVID-19.Purpose:The occurrence of SAT many weeks after administering the second COVID-19 vaccine is rare and has limiteddocumentation in academic literature. This study aims to present the occurrence of SAT after administering the COVID-19vaccine. We present the case of a 37-year-old man who developed SAT 23 days after receiving the second dose of PfizerBioNTech’s COVID-19 mRNA vaccine.Research design and study sample:Due to neck pain and an elevated body temperature (up to 38.2°C), a 37-year-old male subject presented for examination 23 days after receiving the second Pfizer BioNTech mRNA vaccineagainst SARS-CoV-2 viral infection. The patient deniedever having an autoimmune disease or any other disease.Painful neck palpation and afirm, slightly enlarged thyroid gland with no surrounding lymphadenopathy wereidentified during the exam. The heart rate was 104 beatsper minute. All of the remaining physicalfindings werenormal.Data collection and/or Analysis:Data collected during the disease are integral to the medical record.Results:Hematology and biochemistry analyses at the initial and follow-up visits revealed minor leukocytosis, normocyticanaemia, and thrombocytosis, followed by a mild increase in lactate dehydrogenase and decreased iron levels. The patient’sthyroid function and morphology had recovered entirely from post-vaccine SAT.Conclusions: Results from this study emphasise the need for healthcare professionals to promptly report any case of SATrelated to COVID-19 vaccination. Further investigation is warranted to understand the immunopathogenesis of COVID-19-associated thyroiditis and the impact of COVID-19 immunization on this condition

Effects of IGF-1 on the IGFB proteins level in the serum of obese male rats

Insulin-like growth factor binding (IGFB) proteins are a group of six highly conserved proteins that bind to insulin-like growth factors (IGFs) and transport them through the bloodstream to the target cells. The IGFB proteins play an important role in regulating IGF signaling pathways by lengthening their half-life. Obesity and associated disorders are linked to abnormal levels of IGFB proteins. Thus, this study aimed to evaluate how IGF-1 affects the levels of IGFB proteins in the serum of obese rats

Parametar oksidativnog stresa i enzimi antioksidativne zaštite kod gojaznih osoba u Srbiji

Rad je koncipiran, dizajniran i urađen u Laborastoriji za radiobiologiju i molekularnu genetiku Instituta za nuklearne nauke “Vinča” – Institutom od nacionalnog značaja – Univerziteta u Beogradu u saradnji sa Medicinskim fakultetom Univerziteta u Novom Sadu i Univerzitetskim Kliničkim CentromVojvodine. Uvod i cilj. Reaktivne vrste kiseonika (ROS) imaju mogućnost da reaguju sa biomolekulima ćelija i telesnih tečnosti i da ih menjaju. Oksidativni stres (OxS) nastaje u ćelijskim sistemima u uslovima narušene ravnoteže između stepena produkcije i otklanjanja visokoreaktivnih molekula, odnosno, kada produkcija ROS prevazilazi antioksidativne kapacitete datih sistema. Jedan od parametara OxS je 4-hidroksi 2-nonelan (4-HNE), čija koncentracija predstavlja stepen lipidne peroksidacije. Antioksidativni zaštitni sistem (AOS) nastao je tokom procesa evolucije u aerobnim uslovima kao odgovor na toksično delovanje kiseonika. Postoji više nivoa AOS, kao što su enzimski antioksidanti i neenzimski antioksidanti. Među najznačajnijim komponentama enzimskog AOS su superoksid dismutaza (SOD), katalaza (CAT), glutation peroksidaza (GPx) glutation reduktaza (GR). Cilj rada je bio utvrđivanje promena koncentracije parametra OxS i aktivnosti enzima AOS kod gojaznih ispitanika u odnosu na normalno uhranjene ispitanike. Metode. U studiju je bilo uključeno 67 osoba oba pola, od čega 36 normalno uhranjenih osoba (kontrole) i 31 gojazna osoba. Ukupni antioksidativni status (TAS) test je meren primenom Randox TAS kita sa Troloksom kao ekvivalentnim standardom na novoj generaciji Daytona (RX) automatskom hemijskom analizatoru prema uputstvu Randox Co. Za određivanje aktivnosti SOD, GPx i GR u serumu ispitanika korišćene su kolorimetrijske metode i komercijalno dostupni Randox kitovi (Randox Labs, Crumlin, UK). Za određivanje aktivnosti CAT i za određivanje koncentracije 4-HNE u serumu ispitanika korišćeni su komercijalno dostupni OxiSelect kitovi (Cell Biolabs, Inc., San Diego, USA). Rezultati. Dobijeni rezultati pokazuju da je koncentracija 4-HNE kod gojaznih osoba bila statistički značajno viša za 36% (p<0,001) u odnosu na nivo 4-HNE merenog kod kontrola. Nivo TAS kod gojaznih ispitanika bio je statistički značajno smanjen (p<0,001) u poređenju sa vrednostima za TAS kod kontrolnih ispitanika. Procenat smanjenja aktivnosti enzima AOS kod gojaznih osoba bio je 35% za SOD (p<0,001), 21% za GR (p<0,001) i 29% za GPx (p<0,001). Nije uočena statistički značajna promena za aktivnost CAT između dve ispitivane grupe. Zaključak. Dobijeni rezultati jasno pokazuju da stanje gojaznosti dovodi do smanjenja aktivnosti AOS, kao i povećanja koncentracije markera lipidne peroksidacije. Takođe, dobijeni rezultati sugerišu da bi određivanje enzima AOS i markera lipidne peroksidacije mogli biti jedni od biomarkera predikcije nastanka gojaznosti.KES2022 : 8. Kongres endokrinologa Srbije sa međunarodnim učešćem, Nov 30 - Dec 3, 2022, Beogra

The X Files: “The Mystery of X Chromosome Instability in Alzheimer’s Disease”

Alzheimer’s disease (AD) is a neurodegenerative disease that affects millions of individuals worldwide and can occur relatively early or later in life. It is well known that genetic components, such as the amyloid precursor protein gene on chromosome 21, are fundamental in early-onset AD (EOAD). To date, however, only the apolipoprotein E4 (ApoE4) gene has been proved to be a genetic risk factor for late-onset AD (LOAD). In recent years, despite the hypothesis that many additional unidentified genes are likely to play a role in AD development, it is surprising that additional gene polymorphisms associated with LOAD have failed to come to light. In this review, we examine the role of X chromosome epigenetics and, based upon GWAS studies, the PCDHX11 gene. Furthermore, we explore other genetic risk factors of AD that involve X-chromosome epigenetics

Biomarkers of cardiovascular diseases

Biomarkeri predstavljaju indikatore normalnih bioloških procesa, patogenih procesa ili farmakoloških odgovora na terapijske intervencije. Interleukin-6 (IL6, engl. Interleukin-6) je biomarker, čija sinteza može biti aktivirana različitim stimulusima, kao što su: interferon-g (IFN-g, engl. Interferon-g), faktor tumorske nekroze (TNF, engl. Tumor Necrosis Factor) i/ili interleukin-1 (IL-1, engl. Interleukin-1). IL-6 svoje efekte ostvaruje preko IL-6 receptora (IL-6R, engl. IL-6 Receptor). Pokazano je da kod transgenih miševa, kod kojih je indukovana ekspresija IL-6 i IL-6R, dolazi do hipertrofije miokarda. U mehanizmu hipertrofije miokarda bitnu ulogu ima i novootkriveni kardiotrofin1 (CT-1, engl. Cardiotrophin-1) koji je jedan od članova IL-6 familije. Aktivnost IL-6 vezuje se za razvoj aneurizme abdominalne aorte (AAA, engl. Abdominal Aortic Aneurysm), zapravo, pokazano je da su aneurizme mesta odakle cirkuliše IL-6, a takođe se smatra da je koncentracija IL-6 u pozitivnoj korelaciji sa dijametrim AAA. C-reaktivni protein (CRP, engl. CReactive Protein) je jedan od mnogobrojnih biomarkera kardiovaskularnih bolesti. Uloga CRP-a je u nastanku i progresiji kardiovaskularnih bolesti. Lokalna produkcija CRP-a od strane glatkih mišićnih i endotelnih ćelija krvnog suda, u velikoj meri utiče na razvoj procesa ateroskleroze. Važnu ulogu u nastanku ateroskleroze, osim CRP-a, ima i oksidovani lipoprotein male gustine (ox-LDL, engl. Oxidized Low Density Lipoprotein). Oksidaciju LDL-a vrše različiti enzimi. Ox-LDL nakon što prođe u intimu krvnog suda indukuje sakupljanje monocita, tj. monociti se prevode u makrofage koji vezuju ox-LDL. Kada se makrofagi napune ox-LDL-om, dolazi do pokretanja signala ćelijske smrti i stvaraju se forme penušavih ćelija koje čine početni deo aterosklerotičnog plaka. Nova saznanja o mehamizmu delovanja kao i uloge biomerkera u nastanku kardiovaskularnih bolesti, svakako će pružiti jednu od mogućnosti prevencije nastanka ovih poremećaja, a takođe i adekvatnu terapiju u lecenju kardiovaskularnih oboljenja, što i jeste jedan od glavnih ciljeva intezivnih istraživanja u oblasti biomarkera. U ovom preglednom članku, opisana su tri biomarkera kardiovaskularnih bolesti: IL-6, CRP i LDL.Biomarkers are indicators of normal biological processes, pathogenic processes or pharmacologic responses to therapeutic interventions. Interleukin-6 (IL - 6) is a biomarker whose synthesis could be activated by various stimuli, such as interferon-g (IFN - g), tumor necrosis factor (TNF) and/or interleukin - 1 (IL - 1). IL - 6 achieves its effects through the IL-6 receptor (IL - 6R). It has been shown that transgenic mice, which have induced expression of IL - 6 and IL - 6R develop myocardial hypertrophy. In myocardial hypertrophy, an important role is played by a newly discovered cardiotrophin-1, a member of the IL - 6 family. The activity of IL - 6 is associated with the development of abdominal aortic aneurysm (AAA); in fact, it has been shown that the concentration of IL - 6 positively correlates with AAA diameters. C-reactive protein (CRP) is one of the biomarkers of cardiovascular diseases. Local production of CRP by the smooth muscular and endothelial cells of the vessel leads to the development of atherosclerosis to a large extent. Oxidized low-density lipoprotein (ox - LDL) also has an important role in the development of atherosclerosis. After penetrating the intima of the vessel, ox - LDL induces monocyte collection, i.e. monocytes are translated into macrophages that bind ox - LDL. Having filled the macrophages with ox - LDL, the signals of cell death are activated, which leads to the creation of foamy cells that make up the initial part of the atherosclerotic plaque. New knowledge about the mechanism of action and the role of biomarkers in the development of cardiovascular diseases will certainly provide an opportunity to prevent the onset of these disorders, as well as an adequate therapy in the treatment of cardiovascular diseases, which is one of the main goals of intensive research in the field of biomarkers

Regulation of nitric oxide production in hypothyroidism

Hypothyroidism is a common endocrine disorder that predominantly occurs in females. It is associated with an increased risk of cardiovascular diseases (CVD), but the molecular mechanism is not known. Disturbance in lipid metabolism, the regulation of oxidative stress, and inflammation characterize the progression of subclinical hypothyroidism. The initiation and progression of endothelial dysfunction also exhibit these changes, which is the initial step in developing CVD. Animal and human studies highlight the critical role of nitric oxide (NO) as a reliable biomarker for cardiovascular risk in subclinical and clinical hypothyroidism. In this review, we summarize the recent literature findings associated with NO production by the thyroid hormones in both physiological and pathophysiological conditions. We also discuss the levothyroxine treatment effect on serum NO levels in hypothyroid patients. © 2020 The Author

Effects of Levothyroxine Replacement Therapy on Parameters of Metabolic Syndrome and Atherosclerosis in Hypothyroid Patients: A Prospective Pilot Study

The aim of this study was to investigate the effect of levothyroxine (LT4) replacement therapy during three months on some parameters of metabolic syndrome and atherosclerosis in patients with increased thyroid-stimulating hormone (TSH) level. This study included a group of 30 female patients with TSH level GT 4mIU/L and 15 matched healthy controls. Intima media complex thickness (IMCT) and peak systolic flow velocity (PSFV) of superficial femoral artery were determined by Color Doppler scan. In hypothyroid subjects, BMI, SBP, DBP, and TSH were significantly increased versus controls and decreased after LT4 administration. FT4 was significantly lower in hypothyroid subjects compared with controls and significantly higher by treatment. TC, Tg, HDLC, and LDL-C were similar to controls at baseline but TC and LDL-C were significantly decreased by LH4 treatment. IMCT was significantly increased versus controls at baseline and significantly reduced by treatment. PSFV was similar to controls at baseline and significantly decreased on treatment. In this study, we have demonstrated the effects of LT4 replacement therapy during three months of treatment on correction of risk factors of metabolic syndrome and atherosclerosis

Fundamentals of apoptosis

Apoptoza je evolutivno očuvan mehanizam programirane ćelijske smrti, koji ima važnu ulogu u fiziološkim procesima, kao što su embrionalno razviće, hromonima regulisana ćelijska smrt, funkcionisanje imunog sistema i uklanjanje oštećenih ćelija. Dva osnovna puta indukcije apoptoze su unutrašnji i spoljašnji. Dok unutrašnji put apoptoze može pokrenuti unutarćeljska akumulacija Ca 2+ jona, koja je praćena permeabilizacijom membrane mitohondrija i oslobađanjem pro-apoptotskih proteina iz mitohondrija u citoplazmu, spoljašnji put uključuje aktivaciju membransih receptora za TNF-a (engl. Tumor Necrosis Factor-a), kao što su TNFR-1 (engl. Tumor Necrosis Factor receptor 1), Fas, i TRAIL-R (engl. TNF-related apoptosis-inducing ligand receptors). Pored toga, postoji i perforin-granzim put koji uključuje aktivaciju citotoksičnih T-limfocita. Sva tri puta rezultiraju fragmentacijom DNK, degradacijom citoskeleta i nuklearnih proteina, unakrsnim povezivanjem proteina, formiranjem apoptotskih tela, ekspresijom liganada za receptore na fagocitima i na kraju fagocitozom. U ovoj reviji su objedinjeni podaci iz nedavno objavljenih studija koje su fokusirane na proteine uključene u proces apoptoze i molekularne mehanizme apoptoze. Razumevanje mehanizama apoptoze može da pruži korisne informacije i nove pristupe u prevenciji i razvoju novih terapija za različite bolesti.Apoptosis is evolutionary conserved, programmed pattern of cell death with an essential role in various physiological processes, such as normal cell turnover and embryonic development, hormone-regulated cell demise, aging, immune system functioning and development and removal of defective and harmful cells. There are two general pathways for activation of apoptosis: the intrinsic and extrinsic pathways. While the intrinsic apoptotic pathway can be triggered by a cytotoxic accumulation of intracellular Ca 2+ , followed permeabilization of mitochondrial membrane and release of pro-apoptotic proteins into the cytosol from mitochondria, the extrinsic mechanisms of apoptosis include the participation of death receptors of tumor necrosis factor-a (TNF-a), receptor superfamily such as TNFR-1, Fas, and TNF-related apoptosis-inducing ligand receptors (TRAIL-R) located on the plasma membrane. There is also the perforin-granzyme pathway that involves T-cell mediated cytotoxicity. All three pathways converge on the same execution pathway, resulting in DNA fragmentation, degradation of cytoskeletal and nuclear proteins, cross-linking of proteins, formation of apoptotic bodies, expression of ligands for phagocytic cell receptors and finally uptake by phagocytic cells. In this review we summarize data from recent studies focusing on apoptotic proteins that have been identified and molecular mechanisms of apoptosis. Understanding apoptotic mechanism might provide useful information and a new approach to prevention and development of new therapies for variety of diseases