Background: Vascular calcifications (VC) are increasingly prevalent in patients with chronic kidney disease. This study aimed to assess the incidence of iliac artery calcifications in kidney transplant (KT) patients and explore the relationship between iliac VC burden measured by pelvic calcification score (PCS) and renal transplant outcomes. Methods: This prospective study involved 79 KT recipients. VC quantification, using a pre-transplant computed tomography (CT) scan, was performed by assessing calcifications in the common and external iliac arteries bilaterally, resulting in an overall PCS ranging from 0 (no calcifications) to 44 (extensive calcifications). Based on PCS values, patients were divided into three equal-sized groups: PCS Group 1 (PCS 0-4), PCS Group 2 (PCS 5-19), and PCS Group 3 (PCS > 19). Post-transplant outcomes tracked for at least 1 year were patient and graft survival, graft function (urea, creatinine, MAG-3 clearance), and incidence of MACE during the first post-transplant year. Results: Calcifications were present in at least one arterial segment in 61 patients (77.2%). One-year patient survival was 95%, and one-year graft survival was 92.4%. Patients in PCS Group 3 had significantly lower one-year patient and graft survival compared to those in PCS Group 1 and 2 (p = 0.006 and p = 0.008, respectively). MACE and renal function indicators 1-year post-transplant were similar across all PCS groups. Conclusions: Our study demonstrated that a significant majority of KT recipients exhibited iliac VC during pre-transplant CT assessments. Patients in PCS Group 3 exhibited significantly lower one-year patient and graft survival rates compared to those in PCS Groups 1 and 2, indicating that this subgroup may require more intensive post-transplant monitoring and management.

Tetralogy of Fallot is the most common cyanotic congenital heart disease. This severe disorder of cardiac physiology can impair renal function and lead to the development of cardiorenal syndrome and eventually to end-stage renal disease. Kidney transplantation may be the best option for renal replacement treatment in patients with tetralogy of Fallot, but only after correcting cardiac abnormalities and optimizing cardiac functions, all of which require a multidisciplinary approach. We report the first case of kidney transplantation in an adolescent patient with tetralogy of Fallot. Our findings confirms that kidney transplantation is a valuable treatment option in selected congenital heart disease cases.

Chronic kidney disease (CKD) is among the most significant health problems, with the associated cardiovascular disease and bone metabolism disorders being the leading cause of morbidity and mortality in these patients. The aim of the study was to determine markers of bone turnover in patient sera (phosphates, calcium, alkaline phosphatase, parathyroid hormone and osteoprotegerin (OPG)) in all stages of kidney failure including kidney transplant recipients. We also wanted to determine whether dialysis vintage affects recovery of bone markers one year after transplantation. There were 164 study patients, whereas 30 healthy individuals served as a control group. Serum OPG progressively increased with decline of the glomerular filtration rate. The highest OPG concentration was recorded in dialysis group. We observed a statistically significant OPG increase in stage 2 CKD. In kidney transplant group, there was positive correlation between OPG and dialysis vintage. We also found that serum OPG was lower in patients treated with dialysis for less than 4 years prior to transplantation. We confirmed that CKD-mineral and bone disorder began in stage 3 CKD with parathyroid hormone and OPG elevation, and a statistically significant OPG increase in stage 2 CKD might be an early sign of CKD-mineral and bone disorder. Dialysis vintage longer than 4 years is associated with more significant disturbances in mineral and bone metabolism.

Ishemijsko-reperfuzijska ozljeda jedan je od najvažnijih uzroka oslabljene funkcije transplantiranog bubrega. Dosad su identificirani mnogi čimbenici koji mogu povećati vjerojatnost njezina nastanka, a jedan od najvažnijih je način pohrane bubrega, odnosno duljina trajanja hladne ishemije. Osnovni uzrok oštećenja tkiva u hladnoj ishemiji je nedostatak kisika, koji dovodi do smanjenja koncentracije adenozin trifosfata (engl. adenosine triphosphate, ATP-a) te slabljenja aktivnosti Na/K-ATP-aze (Na/K crpke). Puno kompleksniji i teži oblik oštećenja tkiva nastaje ponovnom uspostavom cirkulacije, odnosno reperfuzijom, prilikom koje dolazi do pojačanog ulaska kisika u stanice te stvaranja velike količine slobodnih kisikovih radikala. Njihovo nakupljanje unutar tkiva uzrokuje oštećenja DNA, proteina i lipida stanične membrane. Ljudski organizam djelomično je zaštićen od djelovanja slobodnih kisikovih radikala pomoću vlastitih antioksidativnih enzimatskih sustava, poput superoksid dismutaze (SOD), glutation peroksidaze (GSH-Px) te katalaze. Kako bi se spriječio nastanak navedenih oštećenja, u kliničkoj se praksi za vrijeme trajanja ishemije bubrega koriste različite otopine za njegovo čuvanje.

Cilj: Ciljevi ove studije su istražiti ima li omjer serumskih vrijednosti RANKL/OPG bolje prognostičke vrijednosti uspoređen s razinom iPTH-a u serumu te ispitati ovise li razine OPG-a i RANKL-a u serumu o terapiji vitaminom D. Ispitanici i metode: Istraživanje je obuhvatilo 120 pacijenata iz Kliničkog bolničkog centra Rijeka, Hrvatska. Pacijenti su podijeljeni u dvije skupine; skupina pacijenata na hemodijalizi (HD) i skupina primatelja bubrežnog transplantata. Primatelji transplantata bubrega uključeni su u trenutku transplantacije te praćeni 12 mjeseci nakon transplantacije. Rezultati: Prema rezultatima ovog istraživanja, serumske razine RANKL/OPG u skupini pacijenata na hemodijalizi koji su imali iPTH > 33 pmol/l i iPTH < 11 pmol/l bile su značajno veće u odnosu na one koji su imali iPTH 11 – 33 pmol/l. Također, razine RANKL/OPG u skupini primatelja bubrežnog transplantata koji su imali iPTH > 33 pmol/l i iPTH < 11 pmol/l bile su značajno veće, u usporedbi s onima koji su imali iPTH 11 – 33 pmol/l. Pacijenti na hemodijalizi i primatelji bubrežnog transplantata na terapiji vitaminom D imali su značajno više razine OPG-a u serumu i značajno nižu razinu sRANKL-a, od ispitanika bez terapije vitaminom D. Zaključak: Rezultati ovog istraživanja ukazuju na povoljan učinak vitamina D u prevenciji gubitka koštane mase kod pacijenata na hemodijalizi i kod primatelja bubrežnog transplantata. Omjer serumske razine RANKL/OPG korelira s vrijednostima iPTH-a, ali ostaje nejasno zašto je omjer RANKL/OPG nizak u umjereno povišenim vrijednostima iPTH-a te su potrebna dodatna istraživanja kako bismo dobili odgovor na ovo pitanje.

OBJECTIVE:Restenosis is a frequent complication of angioplasty. It consists of a neointimal hyperplasia resulting from progression and migration of vascular smooth muscle cells (VSMC) into the vessel lumen. microRNA miR-223 has recently been shown to be involved in cardiovascular diseases including atherosclerosis, vascular calcification and arterial thrombosis. In this study, our aim was to assess the impact of miR-223 modulation on restenosis in a rat model of carotid artery after balloon injury. METHODS:The over and down-expression of miR-223 was induced by adenoviral vectors, containing either a pre-miR-223 sequence allowing artificial miR-223 expression or a sponge sequence, trapping the native microRNA, respectively. Restenosis was quantified on stained rat carotid sections. RESULTS:In vitro, three mRNA (Myocyte Enhancer Factor 2C (MEF2C), Ras homolog gene family, member B (RhoB) and Nuclear factor 1 A-type (NFIA)) reported as miR-223 direct targets and known to be implicated in VSMC differentiation and contractility were studied by RT-qPCR. Our findings showed that down-expression of miR-223 significantly reduced neointimal hyperplasia by 44% in carotids, and was associated with a 2-3-fold overexpression of MEF2C, RhoB and NFIA in a murine monocyte macrophage cell line, RAW 264.7 cells. CONCLUSION:Down-regulating miR-223 could be a potential therapeutic approach to prevent restenosis after angioplasty.

Since vascular calcification is considered a process regulated similar to that of bone tissue mineralization, we investigated the participation of bone formation proteins. We analyzed the correlation of serum circulating bone markers, osteoprotegerin (OPG) and receptor activator of nuclear factor ĸB ligand (RANKL) in chronic kidney disease (CKD) patients, to coronary artery calcification score. We also considered the effect of inorganic phosphate on pro- and anti-calcifying tissue factors. We confirmed that circulating OPG is an independent calcium score predictor with its high serum concentration favoring high coronary artery calcification. In tissue samples of non-diseased human renal arteries, the expression of OPG and receptor activator of nuclear factor ĸB (RANK) was positive, while expression of RANKL was absent. In atherosclerotic specimens and arteries with medial calcification, the most upregulated was expression of bone morphogenetic proteins, BMP-2 and BMP-7, as well as expression of RANK and RANKL. In the diseased arteries, OPG expression was present only in areas where bone structures were formed. In atherosclerotic and medial calcification arteries, loss of alpha-smooth muscle actin (α-SMA) expression was observed. These data suggest a possible regulatory role of the examined proteins, especially OPG and RANKL, in vascular calcification, as well as their possible clinical significance as circulating predictors of vascular calcification.

Learning the basic morphology of the brain and topographic relations between certain parts of the brain is fairly complex task in itself and moreover in the Anatomical atlases there is not enough cross‐sectional view of the brain. Therefore, for a successful study of the brain structure through images in the atlas, a great number of images and schemes of coronal, sagittal and horizontal sections are needed. Through a series of such cross‐sections, the student could reconstruct brain structures and study the mutual relationships of neighboring structures.To enable such a way of learning we have made a section of the brain and we recorded it on a CD along with an explanation of all visible structure.We have made a sections of five 10% Formalin‐fixed brains.Two brains were sectioned to show lateral ventricle located within the cerebral hemisphere, third ventricle and fourth ventricle. Then we have made serial 1 cm thick cross‐sections of one brain in coronal plane. Each side of the section was recorded along with an explanation. One brain was cut in median sagittal plane. After that one cerebral hemisphere was cut in horizontal plane in 2 cm thick serial sections while the second cerebral hemisphere was cut in sagittal plane in 2 cm thick serial sections.Live section of the brain is very well accepted by the students and is very helpful for better understanding of the brain morphology.Support or Funding InformationFunded by Faculty of Medicine University of RijekaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Deceased donor kidneys are exposed to cold ischemic insult which makes them particularly susceptible to the effects of cold ischemic injury during hypothermic preservation resulting in high rates of delayed graft function. Bone morphogenetic protein-7 (BMP-7) is a valuable reagent in the field of tissue regeneration and preservation under ischemic conditions. Following these insights, we investigated the effect of recombinant human BMP-7 (rhBMP-7) on graft preservation during cold ischemia. The study was conducted on an experimental model of kidney cold ischemia in rats. Kidneys were perfused with University of Wisconsin (UW) saline solution, rhBMP-7, or rhBMP-7 + UW, and exposed to cold ischemia for 6, 12, and 24 hours. In tubular epithelial cells of kidneys perfused with rhBMP-7 and rhBMP-7+UW solution, the expression of BMP-7 and E-cadherin was observed after 24 hours of cold ischemia. In kidneys not perfused with rhBMP-7, high expression of transforming growth factor-beta and alpha-smooth muscle actin was found. Also, in kidneys perfused with rhBMP-7 solution, statistically higher levels of Smad1, Smad5, and Smad8 messenger RNA expressions were proven. BMP-7 maintains the morphology of kidney tissue better than UW solution during 24 hours of cold ischemia. BMP-7 prevents epithelial to mesenchymal transformation and consequently maintains epithelial phenotype of tubular cells.

Spinal cord ischemia-reperfusion injury (IRI) results in overproduction of reactive oxygen species leading to tissue oxidative stress which impacts the neuronal network in the spinal cord as well as glial cells. We investigated the expression of Nuclear factor erythroid 2-related factor 2 (Nrf2) in neurons and glial cells after occlusion of the abdominal aorta followed by IRI as well as the time-dependent expression of Nrf2 in the same cells. The experimental method of transient aortic occlusion was carried out on rats by cross-clamping of the abdominal aorta for 45 minutes. The animals used for this study were sacrificed 1 h, 6 h, and 48 h after reperfusion to determine time-related changes of Nrf2 expression, as well as changes of astrocyte activity in the spinal cord. Immunofluorescence results showed an increase in the staining intensity of Nrf2 expression in the neurons following ischemia with highest intensity 48 h post-reperfusion and an increase in a number of reactive astrocytes. Western blot analysis showed that Nrf2 protein expression increased in a cytoplasmic and nuclear fraction as early as 1 h after reperfusion and remained active 48 h after, resulting in increased expression of the main Nrf2 target gene HO-1. In conclusion, substances that enhance expression of Nrf2 may have the potential to prevent cellular damage to the spinal cord caused by IRI.

BackgroundSpinal cord ischemia and paralysis of lower extremities are devastating postoperative complications that accompany surgery of aortic aneurysm. Spinal cord ischemia ‐ reperfusion injury (I/R) results in overproduction of reactive species leading to tissue oxidative stress, which impact the neuronal network in spinal cord as well as supporting white matter cells. Here, we report the longitudinal analysis of activity and number of neurologic cells in the area of aortic clamping.MethodsTransient aortic occlusion was produced in rats by cross – clamping of the abdominal aorta for 45 minutes. Animals were sacrificed at several time points after reperfusion; 1, 6 and 48h; to determine time correlated changes in activity and number of neurologic cells in spinal cord. Spinal cord tissue was analyzed by immunofluorescence using specific primary antibodies, glial fibrillary acid protein (GFAP) and NeuN. Also, we wondered whether survival of neurons was related to their expression of nuclear factor erythroid 2‐related factor 2 (Nrf2), an oxidative stress protective protein.ResultsIn the present study, ischemia – reperfusion injury of spinal cord was manifested by moderate loss of neurons in dorsal horn after 6h of reperfusion, and significant loss of neurons in dorsal horn after 48h of reperfusion compared to sham animals. Neurons located ventrally and laterally indicated no change in number. Surviving neuron cells, howerver, showed expression of Nrf2 with its nuclear translocation escpecially pronounced in neurons located in ventral horn. Longitudinal analysis of neuronal number showed that loss of neurons in dorsal horn was limited to lumbar part of spinal cord. Analysis of GFAP positive cells revealed moderate increase in number 6h post reperfusion in lateral part of spinal cord, while 48h post reperfusion this increase was even more pronounced compared to sham animals. Increase of number of GFAP positive cells was followed with astrocyte activation in lateral part of spinal cord presented as increased GFAP positive area in the same part of spinal cord. Longitudinal analysis of spinal cord revealed that astrocyte activation is present even in thoracic parts of spinal cord, higher than segments which were targeted by aortal clamping.ConclusionOur data indicate that I/R spinal cord injury induces changes in number of GFAP and NeuN positive cells and these changes correlate to duration of reperfusion time. The surviving neurons express Nrf2 in their nucleus, which shows the involvement of Nrf2 pathway in neuron survival.Support or Funding InformationGrant approved and financed by Faculty of Medicine University of Rijeka

The discovery of small RNAs has shed new light on microRNA (mRNA) regulation and a range of biological processes. The recognition that miRNAs-221 and -222 are sensitive regulators in the endothelium may enable the identification of novel biomarkers and therapeutic targets. Given that endothelial dysfunction precedes the development of atherosclerosis and contributes to the development of cardiovascular damage, circulating miRNAs produced by Endothelial Cells (ECs) are putative biomarkers for a wide range of cardiovascular diseases. Furthermore, EC proliferation and migration have a critical role in the formation of new blood vessels (angiogenesis), an important component of physiological processes and tumour growth. Hence, the use of anti-angiogenic miRNAs might constitute a novel therapeutic strategy. Along with endothelial angiogenesis, several other processes involving ECs (such as neointimal lesion formation, vascular inflammation, lipoprotein metabolism and hypertension) are critical factors in atherogenesis and atherosclerosis. The fact that human blood-derived progenitor cells, endothelial progenitor cells, umbilical vein ECs and quiescent ECs express high levels of miR-221/222 suggests an important role for this miRNA cluster in endothelial (patho) physiology. The present article reviews current knowledge on miRNAs-221/222's regulation roles in endothelial function and disease in general and angiogenesis in particular.

Structure and shape of the circulatory system differ substantially among animal species, and particularly in some aspects of comparative anatomy they are unrecognizable. Since the circulatory system ensures the functional and morphological integrity of all parts of the organism, its structure depends essentially on the shape of the organism and its biological properties. This paper, in the form of a discussion, presents some aspects of comparative anatomy related to the basic principles of the vertebrate circulatory system.

To verify that co-application of propofol preconditioning and postconditioning protects spinal cord from ischemia/reperfusion injury by enhancing the different subtypes of SOD activity, which is related to PI3K/AKT signal pathway.60 rabbits were randomly equally assigned to 3 groups: Group S, sham-operation group; Group I/R., ischemia/reperfusion group; Group P, ischemia/reperfusion group with propofol treatment. Four rabbits per group were randomly executed at the time-points: days 1, 2, 3, 5, and 7 post-surgery. Spinal cord tissues at L3 to L4 levels were harvested. The bioactivities of SOD1 and SOD2, and the mRNA expression levels of SOD1, SOD2, PI3K, and AKT were detected.On day 1, the bioactivity of SOD1 increased significantly in Group I/R or Group P compared with Group S (P < 0.05). On day 2, compared with Group S, the bioactivity of SOD1 increased significantly in Group P (P < 0.05). On days 3, 5, and 7, the bioactivity of SOD1 decreased significantly respectively in Group I/R compared with Group S (P < 0.05). On all timepoints, the bioactivity of SOD2 decreased significantly in Group I/R compared with Group S (P < 0.05). There was a positive correlation between the SOD1 activity and the respective mRNA expression of SOD1, PI3K, and AKT.Co-application of propofol preconditioning and postconditioning resulted in potent protective effects against spinal cord ischemia/reperfusion injury, which was associated with the increased expression of SOD1 in spinal cord tissues by activating PI3K/AKT signal pathway.

Structure and shape of the circulatory system differ substantially among animal species, and particularly in some aspects of comparative anatomy they are unrecognizable. Since the circulatory system ensures the functional and morphological integrity of all parts of the organism, its structure depends essentially on the shape of the organism and its biological properties. This paper, in the form of a discussion, presents some aspects of comparative anatomy related to the basic principles of the vertebrate circulatory system.

Abstract Herein we have demonstrated that both superoxide dismutase (SOD) mimic, cationic Mn(III) meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin (MnTnHex-2-PyP5+), and non-SOD mimic, anionic Mn(III) meso-tetrakis(4-carboxylatophenyl)porphyrin (MnTBAP3−), protect against oxidative stress caused by spinal cord ischemia/reperfusion via suppression of nuclear factor kappa B (NF-κB) pro-inflammatory pathways. Earlier reports showed that Mn(III) N-alkylpyridylporphyrins were able to prevent the DNA binding of NF-κB in an aqueous system, whereas MnTBAP3− was not. Here, for the first time, in a complex in vivo system—animal model of spinal cord injury—a similar impact of MnTBAP3−, at a dose identical to that of MnTnHex-2-PyP5+, was demonstrated in NF-κB downregulation. Rats were treated subcutaneously at 1.5 mg/kg starting at 30 min before ischemia/reperfusion, and then every 12 h afterward for either 48 h or 7 days. The anti-inflammatory effects of both Mn porphyrins (MnPs) were demonstrated in the spinal cord tissue at both 48 h and 7 days. The downregulation of NF-κB, a major pro-inflammatory signaling protein regulating astrocyte activation, was detected and found to correlate well with the suppression of astrogliosis (as glial fibrillary acidic protein) by both MnPs. The markers of oxidative stress, lipid peroxidation and protein carbonyl formation, were significantly reduced by MnPs. The favorable impact of both MnPs on motor neurons (Tarlov score and inclined plane test) was assessed. No major changes in glutathione peroxidase- and SOD-like activities were demonstrated, which implies that none of the MnPs acted as SOD mimic. Increasing amount of data on the reactivity of MnTBAP3− with reactive nitrogen species (RNS) (.NO/HNO/ONOO−) suggests that RNS/MnTBAP3−-driven modification of NF-κB protein cysteines may be involved in its therapeutic effects. This differs from the therapeutic efficacy of MnTnHex-2-PyP5+ which presumably occurs via reactive oxygen species and relates to NF-κB thiol oxidation; the role of RNS cannot be excluded.