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Hyperbaric oxygenation is the therapeutic or experimental application of 100% oxygen at a pressure level higher than atmospheric pressure. Treatment with hyperbaric oxygen can be a beneficial adjuvant therapy in various disorders characterized by compromised tissue oxygenation and perfusion. However, the effects of hyperbaric oxygenation cannot be simply explained as a compensation of the oxygen deficit. Hyperbaric oxygenation has a much broader influence and has the ability to alter protein expression, modulate signaling pathways and affect vascular structure and function. In addition to the classical indications for hyperbaric oxygen therapy, such as gas gangrene, carbon monoxide poisoning, decompression sickness etc., hyperbaric oxygenation was also shown to have promising potential in the treatment of chronic diabetic wounds. An increasing number of studies support the benefits of hyperbaric oxygen for enhancing wound healing and decreasing the likelihood of negative events such as amputation. Such clinical and experimental evidence, as well as potential mechanisms of beneficial effects with focus on vascular function will be discussed. Additionally, the lack of complete information and the lack of a sufficient number of well- designed research studies on this topic will also be addressed.

Questions: There is a paucity of data on the effect of high salt diet on cerebral vessels dilation mechanisms in response to various stimuli in humans. Present study aimed to evaluate the effect of HS diet on middle cerebral artery (MCA) reactivity to breath holding test (BHT) in healthy human. Methods: Healthy women (N=6) and men (N=5) age 20±1 were included in the study. 3x measurements were performed: 1) before any diet intervention, 2) after 1 week of low salt (LS) diet (< 2, 3g kitchen salt/day) and 3) after 2 week of HS diet (>11, 2 g kitchen salt/day). Transcranial doppler signal (TCD) of the MCA in response to BHT (in duration of 30 seconds) was recorded. Blood pressure was recorded simultaneously by Finopres. Results: Diastolic cerebral blood flow (dCBF) and velocity time integral (VTI) were significantly reduced after HS diet compared to the first measurement. R-R interval, heart rate, systolic cerebral blood flow, cerebral perfusion pressure, cerebral vasomotor reactivity and cerebral pulsatility index did not show statistically significant difference. HS diet did not significantly affect any of arterial blood pressure values. Increased 24h-urinary Na+ excretion after HS diet confirmed that subjects conformed to diet. Conclusions: One week of HS diet can impair the cerebrovascular reactivity in human, even without affecting blood pressure values. Support: University of Osijek research grant, INGI- 2015-16

Hyperbaric oxygenation (HBO2) has become widely accepted adjuvant therapy for various conditions with deprived tissue oxygenation, such as diabetic foot and ulcers, ischemic injuries, sepsis, skin flap transplantations. Besides its beneficiary effects on the tissue perfusion, HBO2 exhibits high toxicity at higher pressures, due to increased oxidative stress and barotrauma. Mechanisms of beneficiary as well as pathophysiological effects of HBO2 on vascular function are intensively investigated and include studies in macrovessels, as well as in microvessels, in different animal models and in humans. For example, our studies in diabetic rats demonstrated significantly reduced dilation of cerebral resistance vessels in response to acetylcholine, which is restored with HBO2, possibly mediated by effect of CYP450- epoxygenases’ vasodilating metabolites EETs on KATP channels. Similarly, in aortic ring studies, it appears that HBO2 enhances NO production and EETs production while oxidative stress was not increased. Intermittent HBO2 shifts the arachidonic acid metabolites production / sensitivity demonstrated in genetically modified rats, diabetic rats and in human. Several enzymatic pathways, most prominently CYP450 hydroxylases and epoxygenases are affected by HBO2 ; this occurs in healthy as well as in diseased vasculature. HBO2 changes vascular function by changing mRNA and protein expression, and subsequently production of mediators, such as EETs and NO. In the model of experimental stroke in diabetic female rats, HBO2 exhibited beneficial effects by reducing brain infarct size. Contrary to intermittent HBO2, acute hyperbaric oxygenation immediately after exposure impairs endothelial dependent vasorelaxation (ACh), NO- dependent which is temporary, reversible and caused by increased superoxide levels. This has been proved on functional level (restored relaxation after TEMPOL in vitro) and by direct measurements of released superoxide in aortic tissue. There is a delay in antioxidative enzyme expression in acute HBO2 treatment which may contribute to impaired vasorelaxation.

U predavanju je prikazana uloga atorvastatina kao farmakoterapijske mjere u prevenciji kardiovaskularnih bolesti te kliničke smjernice koje su proizašle iz spoznaja dobivenih kliničkim ispitivanjima atorvastatina

Introduction: Our previous study showed that acute exposure to hyperbaric oxygen (HBO2) increase oxidative stress parameter leading to impaired vasorelaxation to acetlycholine (ACh), while it was preserved in 4 days intermittent HBO2 protocols (1). Vasodilation in response to hypoxia is mainly mediated by cyclooxygenase (COX) activation and production of prostacyclin (PGI2) (2, 3). However, cytochrome P450- epoxygenase metabolites may contribute to vasodilation in healthy vessels (4). It is known that changes in pO2 may affect the synthesis of metabolites of arachidonic acid (5). The aim of this study was to examine if acute single HBO2 exposure affects the reactivity of isolated rat aortic rings to hypoxia and to investigate if there are changes in vasorelaxation mechanisms due to HBO2. Methods: Healthy 9-12 weeks old Sprague-Dawley rats were divided in 3 groups. Control group (n=18, no treatment), A-HBO2 (n=16, acute HBO2 group exposed to HBO2 single 120’ session of 100% O2 at 2.0 bars with additionally 15’ for gradual compression and decompression) and A- HBO2+24H (n=19, rats tested 24 h after HBO2 exposure). Prior to decapitation and aorta preparation, rats were anesthetized with ketamin- chlorid (75 mg/kg) and midazolam (0.5 mg/kg). Response to reduced pO2 (bath gas mixture from 95% O2, 5% CO2 switched to bath gas mixture of 95% N2, 5% CO2) were tested in norepinephrine-precontracted aortic rings in the absence or in the presence of the NOS inhibitor L-NAME, COX-1, 2 inhibitor indomethacin (INDO), MS-PPOH and superoxide scavenger TEMPOL in tissue bath. Thiobarbituric Acid Reactive Substances (TBARS) assay was used to determine oxidative stress in arterial blood samples. To test differences among groups One- way ANOVA or student t-test was used when appropriate (SigmaPlot v11.2, Systat Software, Chicago, USA), p

Objective: Hyperbaric oxygen (HBO2) treatment, used for a number of pathologic conditions, has incompletely elucidated mechanisms of action in vasculature. Previously, repetitive treatment with HBO2 was found to potentially affect components of the renin-angiotensin system, with HBO2 increasing aortic vascular reactivity to angiotensin-(1–7) in healthy and diabetic rats, while not changing angiotensin-(1–7) serum concentrations. The aim of this study was to investigate whether HBO2 has an effect on serum angiotensin-converting enzyme (ACE) activity. Design and Method: Healthy male Sprague-Dawley rats and rats with streptozocin-induced diabetes mellitus were divided into control and HBO2 groups. The latter were treated with 100% oxygen at a pressure of 2 bars in a hyperbaric chamber for 2 hours a day (with additional 15 minutes for each compression and decompression) for 4 consecutive days. On the fifth day, the animals were anesthetized (2.5 mg/kg midazolam + 75 mg/kg ketamine intraperitoneally) and decapitated. Serum samples were stored at -80°C and quantitative determination of ACE was performed spectrophotometrically using Thermo Scientific Infinity ACE Liquid Stable Reagent. Student's t-test (the samples showed normal distribution) was used for comparison between control and HBO2 ACE levels. Results: Mean ACE serum activity in healthy control rats (n = 10) was 107.660 ± 6.931 U/L, whereas the HBO2 treated healthy group (n = 10) had a mean serum ACE activity of 125.385 ± 18.348 U/L, the difference was statistically significant (t-test, P = 0.010). Mean ACE serum activity levels were 142.150 ± 22.555 U/L in diabetic control (untreated) rats (n = 10) and 136.670 ± 24.024 U/L in diabetic HBO2 rats (n = 10), with no significant difference between these two groups (t-test, P = 0.605). Conclusions: The results suggest that intermittent exposure to HBO2 leads to an increase in serum ACE activity in healthy rats. Such an effect was not observed in diabetic rats treated with HBO2. This data might significantly contribute to better understanding of circulatory HBO2 mechanisms of action in different conditions.

Introduction: Results of our previous studies suggest that acute exposure to hyperbaric oxygen (HBO2) increases oxidative stress parameters (1). It is known that oxidative stress impairs vascular relaxation mechanisms in high dietary salt intake or hypertension. On the other hand, chronic, intermittent HBO2 exposure has beneficiary effect on vascular relaxation in diabetic animals (2). The synthesis of metabolites of arachidonic acid (AA) may be changed due to oxygen partial pressure alteration. Aim of this study was to examine the acute, single HBO2 exposure effects on the vascular reactivity to acetylcholine (ACh) in healthy male Sprague-Dawley rats. Methods: Fifty healthy Sprague-Dawley 9-12 weeks old rats were divided in two groups (25 rats per group) - control group and A-HBOT (acute HBO2 in a single 120 minutes session of 100% O2 at 2.0 bars with additionally 15 minutes for gradual compression and decompression). Response to ACh was studied in norepinephrine precontracted aortic rings (Isolated Organ Bath (Experimetria LTD) with or without L-NAME (NOS inhibitor), indomethacine (non-selective COX inhibitor, INDO), SOD mimetic TEMPOL or MS-PPOH (selective inhibitor of CYP450-epoxygenase that catalyses EETs production). Spectrophotometric method TBARS (Thiobarbituric Acid Reactive Substances) was used for measuring products of lipid peroxidation as a direct indicator of oxidative stress in arterial blood serum samples. Results: Acute exposure of healthy animals to HBO2 leads to the significantly impaired vasorelaxation response to ACh compared to control group. TEMPOL (10-5M) in vitro leads to recovery of vasorelaxation in response to ACh in A-HBOT group. In tested groups of rats, ACh induced vasorelaxation was mediated mainly by NO, with the contribution of CYP450 vasodilator metabolites in A-HBOT group respectively. Measuring of oxidative stress parameters (TBARS) showed significantly increased oxidative stress in A- HBOT group compared to control. Discussion: These results are in concordance with our previous study related to hyperbaric oxygenation. Increased oxidative stress in acute hyperbaric oxygen exposure and oxygen partial pressure changes were showed by Drenjancevic et.al 2013. Conclusion: Oxidative stress is increased in acute exposure to HBO2, which is probably the cause of dilation impairment. Hyperbaric oxygenation may lead to relaxation mechanism alteration mainly by activating CYP450 epoxygenation reactions and synthesis of EETs.

HBO kao terapijska metoda ima važnu primarnu ulogu za određena specifična patološka stanja (kao što su dekompresijska bolest, plinska embolija, otrovanje ugljikovim monoksidom i dr.), dok su novija istraživanja posljednjih desetljeća ukazala na potencijalno koristan učinak HBO u drugim bolestima u kojima pak HBO može imati ulogu kao dodatna (adjuvantna) terapijska mjera i ne zamjenjuje osnovne etablirane terapijske modalitete za pojedine bolesti.

Cilj istraživanja bio je istražiti modulira li hiperbarična oksigenacija (HBO) vaskularnu reaktivnost na angiotenzin II (ANG II) i angiotenzin-(1-7) (ANG-(1-7)) u zdravih i dijabetičnih štakora te ispitati ulogu epoksieikozatrienoičnih kiselina (EETs) i CYP enzima u modulaciji vaskularne reaktivnosti putem HBO. Također, cilj je bio provjeriti imaju li eventualne promjene arterijskog tlaka ili oksidativnog stresa ulogu u modulaciji vaskularnih odgovora. Istraživanje je provođeno ispitivanjem vaskularnih odgovora izoliranih torakalnih aortalnih prstenova (funkcionalne studije) na ANG II i ANG-(1-7) u zdravih ili dijabetičnih štakora (tretiranih protokolom s intermitentnom HBO u odnosu na netretirane), te uporabom visoko selektivnog inhibitora sinteze EETs (MS-PPOH) radi funkcionalnog ispitivanja uloge EETs. Nadalje, provedene su in vivo studije utjecaja HBO na arterijski tlak, te molekularne studije utjecaja HBO na pokazatelje oksidativnog stresa, serumske koncentracije ANG-(1-7) (ELISA), mRNA ekspresije CYP enzima (kvantitativna PCR) i proteinske ekspresije CYP enzima (Western blot) u zdravih i dijabetičnih štakora. Pomoću streptozocina izazvan je animalni model dijabetesa tipa 1. u Sprague-Dawley štakora. HBO nije značajno mijenjala aortalnu reaktivnost na sam ANG II u zdravih i dijabetičnih štakora, ali je vaskularni odgovor na kombinaciju ANG II+ANG-(1-7) bio značajno niži u odnosu n ANG II kontrakciju u zdravih HBO štakora, dok takve razlike nije bilo u netretiranih štakora. HBO je značajno pojačavao aortalni odgovor na sam ANG-(1-7) (nakon prekontrakcije noradrenalinom) u zdravih i dijabetičnih životinja. MS-PPOH poništio je sve učinke HBO na facilitaciju vaskularnih odgovora na ANG-(1-7). Nije bilo značajnih promjena arterijskog tlaka, oksidativnog stresa ili serumske razine ANG-(1-7). HBO je imao tendenciju povećavati ekspresiju CYP enzima, a značajno povećanje mRNA ekspresije izmjereno je za CYP4A1 u zdravih i CYP2J3 u dijabetičnih. Proteinska ekspresija bila značajno povećana za CYP2C11 u dijabetičnih štakora. HBO značajno pojačava vaskularni odgovor na ANG-(1-7), što može objasniti njezine pozitivne učinke u patološkim stanjima. EETs imaju važnu ulogu u posredovanju ove modulacije. HBO povećava ekspresiju pojedinih CYP izoformi, a moguće je da povećava i osjetljivost žile na EETs. Promjene vaskularne reaktivnosti nisu posljedica promjena arterijskog tlaka, oksidativnog stresa ili razine ANG-(1-7). HBO ne mijenja značajno reaktivnost na ANG II u zdravih niti u dijabetičnih štakora.

Objective To investigate whether hyperbaric oxygenation (HBO) modulates vascular reactivity to angiotensin II (ANG II) and angiotensin-(1- 7) (ANG-(1-7)) in healthy and diabetic rats and to assess the role of epoxyeicosatrienoic acids (EETs) and CYP enzymes in the modulation of vascular reactivity by HBO. Design and Methods Isolated aortic ring preparations were used to test vascular reactivity (functional studies) to ANG II and ANG-(1-7) in healthy and diabetic rats (treated with an intermittent HBO protocol compared to nontreated), and a selective inhibitor of EETs synthesis (MS-PPOH) was used to assess the functional role of EETs. In vivo studies of HBO effects on arterial blood pressure were conducted, as well as molecular studies of the effects of HBO on oxidative stress indicators, serum concentration of ANG- (1-7), mRNA expression (qPCR) and protein expression (Western blot) of CYP enzymes in healthy and diabetic rats. Results HBO did not significantly change aortic reactivity to ANG II in healthy and diabetic rats. Vascular responses to the combination of ANG II + ANG- (1-7) were lower compared to ANG II contractions in healthy HBO rats, whereas such a difference was not present in untreated rats. HBO significantly increased aortic responses to ANG-(1-7) alone in healthy and diabetic animals. MS-PPOH reversed all effects of HBO on facilitation of vascular responses to ANG-(1- 7). There were no significant changes in arterial pressure, oxidative stress and serum ANG-(1-7) levels. HBO significantly increased mRNA expression of CY4A1 in healthy and CYP2J3 in diabetic rats, while CYP2C11 protein expression was significantly increased by HBO in diabetic rats. Conclusion HBO facilitates responses to ANG-(1- 7), while EETs seem to play an important role in the mechanism of this modulation. HBO increases the expression of specific CYP isoforms. The changes in vascular reactivity were not a consequence of changes in arterial pressure, oxidative stress or ANG-(1- 7) blood levels.

Inflammatory bowel disease (IBD), encompassing ulcerative colitis (UC) and Crohn's disease (CD), is an uncontrolled chronic inflammation of the gastrointestinal tract caused by an interaction of diverse genes and environmental factors. There is growing evidence that cytokine production plays an important role in IBD. One of the key roles in signaling pathway in development of IBD is performed by interleukin 6 (IL-6), although molecular mechanism of this pathway is not yet fully understood. In order to assess the clinical relevance of IL-6 serum concentration in patients with CD and UC we performed cross-sectional, case-control study of IL-6 levels in patients' and healthy blood donors' sera. A total of 100 CD and UC patients and 71 healthy blood donors were investigated. Clinical activity of CD and UC was evaluated using the Crohn's disease activity index and Truelove-Witt's criteria, respectively. Quantitative assessment of serum IL-6 was performed with solid-phase, enzyme-labeled, chemiluminescent sequential immunometric assay. Our results indicate that serum IL-6 is a clinically relevant parameter for CD and UC that strongly correlates with inflammatory activity of disease. We confirmed and extended the role of cytokine production patterns for IBD presentation in Croatian population.

OBJECTIVE: In previous studies, we found that hyperbaric oxygenation (HBO) influences vascular reactivity, possibly through upregulation of epoxyeicosatrienoic acids (EETs). EETs, produced by specific epoxygenase enzymes, have a protective role in many conditions including atherosclerosis and hypertension. Impairment of this role of EETs contributes to endothelial dysfunction in hypertension and increase in EETs production attenuates abnormal renal function. Infusion of EETs decreases mean arterial pressure (MAP) in rats. The aim in this study was to evaluate the effect of HBO on epoxygenase protein expression and blood pressure in healthy or streptozocin- induced diabetic male Sprague-Dawley rats. DESIGN AND METHOD: Rats were divided into control and HBO groups. The HBO group was exposed to 100%O2 for 2 hours (with additional 15 minutes for compression/decompression) daily for 4 consecutive days. On the fifth day, anesthetized animals were sacrificed and aorta samples stored at -80ºC. Western blot analysis was performed with specific antibodies for rat epoxygenases (cytochrome P450 isoforms CYP2J3 and CYP2C11). Epoxygenase expression was quantified relatively to ß-actin expression. MAP was measured in separate anesthethized control and HBO animals after insertion of a femoral artery catheter. T-test or Mann Whitney U test was used to compare HBO and control values, depending on distribution normality. RESULTS: Relative CYP2J3/ß-actin expression was similar between the HBO and control group both in healthy (1.488±1.840 [n=11] vs.1.160±0.785 [n=9]) and in diabetic (1.119±0.495 [n=9] vs. 0.945±0.506 [n=11]) rats. Relative CYP2C11/ß-actin expression was similar between healthy HBO and control animals (1.295±1.031 [n=9] vs.1.425±1.174 [n=8]), but it was statistically significantly higher in the HBO group of diabetic rats compared to control (1.395±0.448 [n=7] vs. 0.796±0.534 [n=7] ; P

The Interleukin-23 signalling pathway, important for the differentiation of TH17 lymphocytes, is involved in the pathogenesis of Inflammatory bowel disease (IBD). Polymorphisms in the IL-23 receptor gene were previously found to be associated with IBD in various populations. The specific rs11209026 and rs7530511 single-nucleotide polymorphisms in the Interleukin-23 receptor gene were determined and associations with Crohn's disease (CD) and ulcerative colitis (UC) were assessed in a Croatian patient population. A total of 50 patients with CD and 93 patients with UC, as well as 99 healthy control subjects were included in the study. In the combined population (IBD, UC+CD), the rs11209026 A allele was differentially distributed between cases and controls (2.1 vs 7.1%, P=0.014), indicating a significant reduction in IBD risk for carriers (allelic OR 0.28 ; 95% CI 0.11-0.75). With respect to each of the populations, the prevalence of the uncommon rs11209026 p.381Gln variant was lower in CD, but not UC patients compared to healthy controls, suggesting a protective effect against CD. Genotype distributions showed that a significantly reduced risk of IBD (OR 0.27, 95% CI 0.1-0.72), limited most likely to the CD phenotype only (MUE 0.09, 95% CI 0-0.54), was observed for heterozygous A/G carriers as compared with the homozygous G/G genotype. No minor allele homozygotes or heterozygotes were observed for CD cases, nor were rare AA homozygotes observed in any other study group. There was no significant association between the rs7530511 polymorphism with either UC or CD. The results suggest a protective effect of the rs11209026 polymorphism for IBD (limited primarily to CD). The investigated associations give potentially important insight into the roles of specific Interleukin-23 receptor polymorphisms in CD pathogenesis in the Croatian population.

The Interleukin-23 signalling pathway is important for the differentiation of TH17 lymphocytes and is involved in the pathogenesis of Inflammatory bowel disease. Polymorphisms in the IL-23 receptor gene were previously found to be associated with Inflammatory bowel disease in various populations. The aim of this study was to determine whether the specific rs11209026 and rs7530511 single-nucleotide polymorphisms in the Interleukin-23 receptor gene are associated with Crohn's disease and ulcerative colitis in a Croatian patient population. A total of 50 patients with Crohn's disease and 93 patients with ulcerative colitis, as well as 99 healthy control subjects were included in the study. The results determined a significantly higher occurrence of rs11209026 in control group compared to patients with inflammatory bowel disease, suggesting a protective effect of this polymorphism. The rs11209026 variant was strongly associated with Crohn's disease, but it was absent in ulcerative colitis. However, there was no significant association between the rs7530511 polymorphism with either ulcerative colitis or Crohn's disease. Associations presented in this study give potentially important insight into the roles of specific Interleukin-23 receptor polymorphisms in Crohn's disease pathogenesis in the Croatian population.

Rad se bavi razvojem simptoma od metaboličkog sindroma do inzulinoma te pristupom bolesniku s metaboličkim sindromom

Metabolic syndrome is most commonly caused by improper, irregular diet, and physical inactivity. However, sometimes the treatment approach to metabolic syndrome is not easy, and if a patient with a low calorie diet and physical activity does not lose weight, other underlying diseases need to be suspected. A sixty-seven year old patient with metabolic syndrome is presented (obesity, accumulation of fat in the abdomen, glucose intolerance, dyslipidemia, and hypertension), who’s attempts at reduction diets and physical activity caused a strong feeling of hunger, general weakness and nausea ; these symptoms were reduced after food intake. Having had a haematemesis (vomiting blood) the patient was admitted to the department of gastroenterology. Examination indicated hiatal hernia (a shift of the upper part of the stomach into the chest through the diaphragm) and chronic gastritis, and for established hypoglycemia with glucose 1.5 to 2.6 mmol/l. The patient was transferred to the department of endocrinology. Additional endocrinologic investigations found insulinoma, a pancreatic tumor sized 0.9 cm. The patient refused surgery and was treated with medication, receiving diazoxide tablets. After two years she had a proper body weight and had no signs of metabolic disorders, and computed tomography of the abdomen showed a significant decrease in pancreatic tumor size. Therefore, in patients with metabolic syndrome in which low calorie diet and physical activity does not reduce body weight, or if they have associated symptoms, endocrinological treatment should be conducted.

Obesity is most commonly caused by improper, irregular diet, and physical inactivity. However, sometimes the treatment of obesity is not easy, and other underlying diseases should be suspected. We present a forty-six year old patient who previously had a proper weight, and with stressful situations at work and in the family had increased appetite resulting in significant weight gain. Once divorced, and after her daughter was placed in the care of the father, the patient was depressed and sought help from a psychiatrist who involved antidepressant therapy. Through the patient’s evaluation, with the obesity and increased waist circumference, it was determined the patient also had diabetes mellitus, dyslipidemia, dominant hypercholesterolemia and hypertension. With the results of the evaluation, the patient was diagnosed with metabolic syndrome. The patient’s neat glycemia, lipids and blood pressure suggested her therapy. With an extreme low calorie diet and physical activity the patient was not losing weight. Endocrinological workup determined hormonal treatment for the presence of hypothyroidism, and a substitution therapy was introduced. After three years, she has a proper body weight, waist circumference, lipids and blood pressure. Medications for diabetes have been significantly reduced. Therefore, in obese patients with metabolic syndrome who have hypercholesterolemia, it is necessary to consider thyroid hormones because using substitution therapy also corrected lipid values and often other components of metabolic syndrome.

The Signal Transducers and Activators of Transcription(STATS) are intracellular effector molecules of cytokine-modulated signalling, which play important role in the development of the human immune system and haematopoiesis, and are involved in the regulation of T-cell survival. STAT3, a master regulator of Th17 and FoxP3+Treg polarization, was recently associated with increased risk of ulcerative colitis (UC) and Chron’s disease (CD). Aim was to investigate whether a functional single nucleotide polymorphism (SNP) in the STAT3 gene is likely to be important for UC and CD risk in a Croatian population, and whether its phenotypic relationship could provide useful clinical predictions. A total of 50 CD patients and 91 UC patients, as well as 95 healthy control subjects were included in the study. The intronic variant in the STAT3 gene (GenBank: NM 213662, rs744166) was genotyped using fluorescence resonance energy transfer technology and melting curve analysis of polymerase chain reaction products. The observed population allele frequencies in the controls were similar to those reported in other Caucasian populations (36.8%, G allele). No significant difference was observed in genotype and allele frequencies between the cases and controls (CD: odds ratio 1.29, 95% confidence interval 0.78–2.11, P = 0.322 ; UC: 0.86 (0.56–1.32), P = 0.502, allelic contrasts, G allele). No further associations were uncovered by inspection of secondary traits. Here, we have demonstrated that the STAT3 rs744166 variant is not associated with CD, UC susceptibility or disease severity in the Croatian population, but pathogenetic mechanisms remain to be further evaluated. The STAT signalling pathway remains a potential therapeutic target for the development of new treatment options for UC and CD.

Hyperbaric oxygenation (HBO2) is widely used clinical treatment for various ischemic injuries. Recently, cellular and molecular mechanisms of HBO2 affecting vascular function are being extensively studied. For example, inhibition of 20-HETE and/or increasing of epoxyeicosatrioneic acids (EETs) levels may decrease cerebral damage following stroke, while levels of these mediators may be modified by HBO2. Our recent studies in rat healthy conductive vessels suggest the role for EETs in modulating relaxation response to ANG-(1- 7) with HBO2 that is mediated via potassium channels other than KATP channels. HBO2 increased the responses to ANG-(1- 7). Furthermore, our studies in diabetic rats showed significantly reduced dilation of cerebral resistance vessels in response to reduced pO2 and ACh, which is restored with HBO2, possibly mediated by effect of CYP450- epoxygenases' vasodilating metabolites of EETs on KATP channels. In addition, our data on mRNA expression showed up-regulation of CYP4A3 and CYP450-epoxygenase genes in cerebral blood vessels in diabetic rats treated with HBO2, and increased protein expression of CYP4A1, CYP4A2 and CYP4A3. The latest two share 97%homology, and both are known to catalyze arachidonic acid ω-hydroxylation and 11, 12-epoxidation thus promoting not only vasoconstriction, but also vasodilation. taken all together, these results demonstrate that HBO2 promotes protection of vascular function in macro- and microcirculation by changeing mRNA and protein expression, and subsequently, restoring dilation responses of vessels in various beds. This improves vessels' function and tissue perfusion by affecting several enzymatic pathways, most prominently CYP450 hydoxylases and epoxygenases, both in healthy and diabetic circulation.

Poster is explaining complexity of approach to patients with Addison's disease or life of Addison's disease

Hipoglikemija je najčešća komplikacija u liječenju bolesnika s dijabetesom. Međutim, ponekad pristup hipoglikemiji nije jednostavan, i potrebno je posumnjati na druge bolesti u podlozi. Prikazan je tridesetogodišnji bolesnik s tipom 1 šećerne bolesti koji je u hitnoj medicinskoj pomoći pregledan zbog teške hipoglikemije i općeg lošeg stanja, te je primio koncentriranu glukozu i infuziju 5% glukoze. Pri dolasku u kliniku, bio je somnolentan, blijed, oznojen, hipotenzivan i bradikardan. Unatrag dva dana imao je mučninu, povraćao je, glavobolju, omaglicu i nedostatak zraka. Ranije je obrađivan kod neurologa zbog kronične demijelinizirajuće polineuropatije. U EKG-u bio je prisutan AV-blok 2. stupnja i inverzni T valovi u inferiornim odvodima. Snimanjem desnih prekordijalnih odvoda i određivanjem kardioselektivnih enzima utvrđen je infarkt desne klijetke. Koronarografijom je potvrđena višežilna bolest sa stenozom desne koronarne arterije te je nakon tri tjedna učinjena kardiokirurška revaskularizacija. Budući da dijabetes uzrokuje neuropatiju, važno je imati na umu da bol u prsima može biti odsutna u akutnom koronarnom sindromu. Prema tome, u dijabetičara s hipoglikemijom kojima se opće stanje ne poboljšava nakon regulacije glikemije ili imaju dodatne tegobe, treba učiniti dodatnu obradu ; u onih koji imaju hipotenziju i bradikardiju svakako treba učiniti i kardiološku obradu.

Poster opisuje pristup bolesniku s debljinom koji u podlozi boluje od hipotireoze.

U radu se kroz prikaz slučaja i teorijsku podlogu prikazuje razvoj kliničke slike od postporođajnog metastatskog tumora do adenoma paratireoidne žlijezde.

Poster is presenting algoritm of approaching hypoglycemia in type 1 diabetes: patient and family education about blood glucose regulation for the prention of acute coronary syndrome

Primary hyperparathyroidism usually presents as hypocalcaemia in an otherwise asymptomatic patient, or alternatively with complications including nephrocalcinosis, osteoporosis or psychoneurologic disorders. However, primary hyperparathyroidism and parathyroid gland adenoma can represent a diagnostic and therapeutic challenge because it is often not recognized at an appropriate time point. A 36 year old female patient is presented who was examined in the Department of Orthopedic Surgery Outpatient Clinic because of pain in her right hip area and lumbar region. Following initial radiologic diagnostics, postpartum advanced metastatic cancer with multiple osteolytic lesions was suspected, due to which she was referred to the Internal medicine clinic for further diagnostic evaluation. After extensive investigation, it was determined that the underlying condition was a parathyroid gland adenoma and the patient was treated surgically with success. The report illustrates the necessity of taking this disease into consideration in differential diagnosis and workup of suspected oncologic patients and describes one of the many faces of parathyroid gland adenoma. Clinicians should note that parathyroid adenoma can mimic metastatic malignant disease, and should be familiar with appropriate diagnostic algorithms and tests that will lead to a timely diagnosis.

Hyperbaric oxygenation (HBO) is the medical use of pure oxygen at a level higher than atmospheric pressure. There are different protocols of HBO being used for research purposes ; however, data on the changes in blood pressure, oxidative stress and acid-base and gas status induced by some of the possible oxygenation patterns/protocols are scarce and conflicting. In this work we aimed to investigate the immediate influence of oxygenation at a pressure of 2 bar during 2 hours on blood pressure, acid- base and blood gas status and parameters of oxidative stress in healthy rats. Healthy male 12-15 week old Sprague-Dawley rats were anesthetized with ketamine (75 mg/kg) and midazolam (2.5 mg/kg) i.p. Femoral artery was cannulated for blood pressure measurements (AP) and blood sampling prior and immediately after single time hyperbaric oxygenation (HBO) in a hyperbaric oxygen chamber (100% oxygen at a pressure of 2 bar for 2 hours, with additional 15 minutes for compression and decompression). Arterial blood samples were collected to analyze acid-base and blood gas status and to determine Ferric reducing ability of plasma (FRAP) and Thiobarbituric Acid Reactive Substances (TBARS ; based on reaction of malonilaldehyde (MDA) with thiobarbituric acid). Paired t-test for AP, blood gases and acid-base status (N of rats = 12) and t- test for FRAP and TBARs (N of rats = 12) were used ; p< ; 0.05 was considered significant. Data are expressed as mean± SD. The study was approved by Ethical Committee of Faculty of Medicine University of Osijek. After HBO there was significant decrease in systolic and diastolic AP (control 138± 14/ 103± 13 vs. 113± 12/ 72± 16 kPa after HBO), although in normal AP range. pH significantly decreased after HBO (control 7.34± 0.05 vs. 7.28± 0.05 after HBO). After HBO, pCO2 was significantly decreased (7.07± 0.89 vs. 5.76± 0.50 kPa) and pO2 significantly increased (12.48± 0.88 vs. 13.68± 2.4 kPa). Plasma bicarbonate significantly decreased (control 27.04± 3.25, vs. 20.52± 3.02 after HBO). Exposure to HBO significantly increased MDA concentration (from 0.173± 0.092 to 21.79± 1.05 microM/MDA), while FRAP was significantly decreased compared to control condition (0.152± 0.035 to 0.117± 0.013 mM/L TE). Control values are in agreement with previously published data on anesthetized rats (1). Acute exposure to HBO had significant effects on all observed parameters. Acidosis may be contributed to prolonged anesthesia (2). However, there is immediate improvement in pCO2 and pO2, although with increase in oxidative stress products and acute decrease in plasma antioxidative capacity, after single time HBO exposure (3).

We previously reported a facilitating effect of hyperbaric oxygen therapy (HBOT) on responses of rat aortic rings to angiotensin-(1-7) (ANG-(1-7)) in healthy rats (‘ Physiology 2011')[1]. It was suggested that this influence was mediated by changes in production or sensitivity to epoxyeicosatrienoic acids (EETs). The aim was to assess whether HBOT exerts similar effects in diabetic rats and to further explore the role of cytochrome P450 (CYP) enzymes in changes induced by HBOT. Diabetes was induced by streptozocin in 6 weeks old male Sprague-Dawley rats. After 8 weeks, they were divided into control and HBOT group (exposed to 100% O2(2 bars) 2 hours/day for 4 consecutive days). Before decapitation, rats were anesthetized with 75 mg/kg ketamine with 2.5 mg/kg midazolam. Thoracic aortic rings were used to test responses to 1 μ M ANG-(1-7) after precontraction with noradrenaline for 5 minutes (tension was read 3 minutes after ANG-(1-7) addition and the response expressed as percentage of precontraction decrease). Intactness of endothelium was previously tested. A separate series of HBOT rats was used to test ANG-(1-7) responses after the epoxygenases inhibitor MS-PPOH (10 μ M) - inhibitor of the CYP4A2 AND CYP4A3 epoxygenation reactions - was added for 15 minutes to the tissue bath. CYP 4A1, 4A2, 4A3 and 2J3 mRNA expression was determined with Quantitative real time PCR and was normalized to the expression of two housekeeping genes -HPRT and 18S. The study was approved by Ethical Committee of Faculty of Medicine University of Osijek. The route of administration of both drugs is intraperitoneal (i.p.). Mean percentage of precontraction decrease after ANG-(1-7) addition was significantly higher in the HBOT group (19.2% ± 7.3 [STDEV]) (n=18) than in the control group (12.1% ± 6.4)(n=14), P= 0.007 (t-test) . The ANG-(1-7) ring response of HBOT rats after MS- PPOH addition was 2.3% ± 3.3 (n=8), significantly less pronounced compared to HBOT rings when no MS-PPOH was used, P< ; 0.001 (Mann Whitney U). Median CYP2J3/18s expression was 12.2 (control)(n=7) and 94.8 (HBOT)(n=6), significantly different (P=0.008, Mann Whitney U). Median CYP2J3/HPRT expression was 7.5 (control)(n=9) and 49.7(HBOT)(n=9), with significant difference (P= 0.042). CYP4A1, CYP4A2 and CYP4A3 expression was similar between groups. HBOT increases the reactivity of thoracic aortic rings to ANG-(1- 7) in diabetic rats. The epoxygenases inhibitor MS-PPOH reverses these changes. HBOT increases mRNA expression of the epoxygenase CYP2J3 in diabetic rats, further suggesting an important role of EETs in HBOT-induced modulation of vascular function.

Hyperbaric oxygen treatment (HBOT) involves exposure to 100% oxygen under high pressure (higher than 1 bar). It is well established treatment in many conditions including wound healing, burnings, multiply sclerosis, gangrene, sepsis and others. The mechanism of hyperbaric oxygen therapy is yet unknown. The aim of this pilot study was to determine the influence of HBOT on expression of several genes important in vasomotor function in aorta and cerebral blood vessels of 14 weeks old healthy male Sprague Dawley rats. Rats were not treated (control) or treated with 100% oxygen pressurized to 2 bars, for two hours consecutively for 4 days. On 5th day rats were sacrificed. The relative expression of mRNA was determined by real time PCR. Results showed significant upregulation of CYP4A1 gene in aortas, and CYP4A3 gene in cerebral blood vessels in treated group, compared to controls. Also, the tendency of increased expression in iNOS, eNOS, COX and CYP450-epoxygenase and other CYP450 hydroxylase genes in aortas and cerebral blood vessels were noticed. The tendency of decreased expression in aortal PGI2 and cerebral blood vessel TA2 mRNA was present. These results suggest that HBOT led to upregulation of genes that are important in production of vasodilators and vasoconstrictors.

Introduction: Hyperbaric oxygen therapy (HBOT) has various effects on perfusion and vascular function and it has been hypothesized that HBOT may change vascular sensitivity to different dilators and constrictors [1]. Angotensin-(1-7) (ANG-(1-7))has been demonstrated to have dilating activity, in contrast to angiotensin II (ANG II) [2] ; both peptides having complex mechanisms of action. The aim of our study was to assess whether there are effects of HBOT on vascular reactivity to ANG II and ANG-(1-7). Methods: Thoracic aortic ring preparations from Sprague-Dawley rats, divided into an HBOT and a control group, were used to test responses to ANG II, ANG II+ ANG-(1-7) or ANG-(1-7) after precontraction with noradrenaline. HBOT rats were treated in a hyperbaric chamber with 100% O2(2 bar) 2 hours/day for 4 consecutive days. After intactness of endothelium was tested with ACh, each ring was subjected to maximal contraction (using 60mM KCl + 0.1μM noradrenaline), and after washing/equilibration treated with either 1 μM ANG II (n[HBOT]=17 ; n[control]=16), 1 μM ANG II+1 μM ANG-(1-7) (n[HBOT]=14 ; n[control]=17), or noradrenaline 0.1 μM - for 5 minutes, after which 1 μM ANG-(1-7) was added and the ring tension read after 3 minutes (n[HBOT]=12 ; n[control]=17). The peak contraction force after ANG II and ANGII + ANG-(1-7) was expressed as percentage of maximal contraction, the effect of ANG-(1-7) addition was expressed as percentage of precontraction decrease after 3 minutes. Results: Mean percentage of maximal contraction for ANGII was 21%±11 (HBOT) and 20%±9 (control) and was similar between groups (Mann-Whitney U test, P=1, 000). The mean percentage for ANGII + ANG-(1-7) was 15% ±10 (HBOT) and 20%±9 (control) ; it tended to be decreased in the HBOT group, but without statistical significance (Mann-Whitney U test, P=0, 054). There was a statistical significance when the mean percentage of maximal contraction of ANGII (HBOT) was compared to ANGII + ANG-(1-7) in the HBOT group (P=0, 029(Mann-Whitney U)), without such a difference within the control group (P=0, 953, t-test). Mean percentage of noradrenaline precontraction decrease with ANG(1-7) was 10%±9 (control) and 19%±11 (HBOT) - significantly different between groups (t-test, P=0, 017). Conclusion: There was no change in reactivity to ANG II after HBOT. There may be an increase of reactivity to ANG-(1-7) after precontraction with noradrenaline, but since there was no significant effect of HBOT on the ANG II + ANG-(1-7) peak compared to control, at least at 1 μM concentrations, further studies are needed to conclusively interpret these results. Interestingly, there is a difference between the peak of ANGII in the HBOT group and ANGII + ANG-(1-7) in the HBOT group, whereas this difference was not present in the control group, suggesting an influence of HBOT on vascular reactivity.

Infection and inflammation are common causes of reactive thrombocytosis. We investigated thrombocytosis in selected patients with inflammatory bowel disease.

One of the possible ways to inhibit development of a tumor is blocking the growth of new blood vessels that provide oxygen and nutrients to the malignant cells. A new approach is to use the high specificity of monoclonal antibodies to target essential angiogenic molecules. These antibodies are derived from cell lines consisting of identical clones, and recognize the same part of a macromolecule. The most relevant are bevacizumab and vitaxin. The first one is approved for the first-line treatment of metastatic colorectal cancer, whereas vitaxin is still in clinical trials. Preliminary results are optimistic, and the drugs are generally well tolerated by the patients.

Transferin je glikoprotein plazme kojeg izlučuju različite stanice u organizmu, prije svega hepatociti. Glavna mu je zadaća održavanje homeostaze željeza, a djeluje i kao čimbenik rasta tijekom embrionalnog razvoja, ali i kod tumora. Veže se s transferinskim receptorima i procesom endocitoze ulazi u stanicu, što se nastoji iskoristiti za dostavu lijekova određenim stanicama. Sustav transferin/transferinski receptor povezan je s poremećajima proizvodnje transferina (atransferinemija, hipotransferinemija) i regulacije željeza (nasljedna hemokromatoza), a vjerojatno i s nekim neurodegenerativnim bolestima.