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T. Jukić, Ana Jurin Martić, S. Ivanković, M. Antica, Doroteja Pavan Jukić, Cecilija Rotim, M. Jurin

SUMMARY The role of T regulatory lymphocytes (Treg) particularly in cancer is well known. The goal of the present study was to determine the contribution of these lymphocytes in the regulation of anti-tumor immunity of CBA/HZgr mice against MC-2 fibrosarcoma (4th generation of methylcholanthrene induced tumor). The levels of T lymphocytes (CD4+, CD8+ and CD4+CD25+) were determined 8 and 20 days after tumor transplantation. Further, the role of CD4+CD25+ (Tregs) in tumor-host interaction was evaluated in vitro and in vivo by using specific monoclonal antibodies. We found that splenocytes of both control and Treg depleted tumor bearing mice strongly but differently inhibited growth of tumor cells in vitro. While splenocytes of untreated mice exhibited significant decrease of this activity (from 74.4% to 62.6% and 32.95%), the splenocytes of Treg depleted mice showed increase of this activity (from 79.5% to 84.3% and 86.2%) from day 6 to day 13 and day 21 after tumor grafting, respectively. Further, upon i.v. injecting specific monoclonal anti-Treg antibody tumor immediately prior to tumor cell intracutaneous transplantation, the tumor was rejected after initial growth. In treated mice, the incidence of Treg cells was very low initially, reaching normal values two weeks later. These animals were shown to be resistant to tumor transplantation four months later.

C. Garofalo, I. Ferrocino, A. Reale, R. Sabbatini, V. Milanović, Mersiha Alkić-Subašić, F. Boscaino, L. Aquilanti et al.

Armin Kasumović, Ines Matoc, N. Avdagić, Tarik Halimić, Bahrija Voloder, L. Muhamedagić, Seldjana Catovic Delic, Irena Sesar

Introduction: To show the importance of optical coherence tomography (OCT) angiography imaging of superficial and deep capillary network in patients with non-proliferative diabetic retinopathy (NPDR), and to show the correlation between blood glucose level and changes in the foveal microvasculature. Methods: A cross-sectional study was performed on eyes with NPDR and healthy subjects using a highspeed 840-nm-wavelength spectral-domain optical coherence tomography instrument (RTVue XR Avanti; Optovue, Inc, Fremont, California, USA). Blood flow was detected using the split-spectrum amplitude-decorrelation angiography algorithm. A fully automated microstructural analysis of the foveal avascular zone (FAZ) and avascular surfaces was performed. Quantitative values from diabetic patients were compared with those of healthy subjects. Data about blood status in diabetic patients and healthy subjects were taken from patients’ medical history. Results: Size of both, FAZ and vascular dropout are significantly different between healthy patients and patients with NPDR. OCT angiography detected enlargement and distortion of the foveal avascular zone, retinal capillary dropout, and a higher number of vascular loops and microaneurysms. Sizes of FAZ and vascular dropout increase with the duration of disease. Central macular thickness (CMT) is not significantly different between healthy patients and patients with NPDR. A study has proven a positive correlation between the size of FAZ and the size of vascular dropout in superficial vascular plexus in patients that have DM over 10 years. Conclusion: A qualitative and quantitative OCT angiography approach to retinal vascular status can offer objective data on monitoring patients with non-proliferative diabetic retinopathy as well as indicate the progression of the disease.

M. Jovanović, M. Petrović, J. Miočinović, S. Zlatanović, Jovanka Laličić Petronijević, D. Mitić-Ćulafić, S. Gorjanović

To meet the demand for new functional foods in line with the trend of sustainable development, a novel probiotic yogurt fortified with 1%, 3%, and 5% apple pomace flour (APF) added immediately after inoculation with Lactobacillus acidophilus, Streptococcus thermophilus, and Bifidobacterium bifidum was developed. Upon fermentation in the presence of APF, a number of probiotic strains remained within the required range, while the syneresis of enriched yogurts was reduced up to 1.8 times in comparison to the control. Supernatants (i.e., extracted whey) obtained from yogurts with 1%, 3%, and 5% APF respectively had 1.4-, 1.8-, and 2.3-fold higher total phenolic content (TPC) than the control, 3.3-, 4.7-, and 8.0-fold higher radical scavenging (DPPH), and 1.3-, 1.6-, and 1.7-fold higher reducing activity (FRAP). Also, probiotic yogurt supernatants (3% and 5%) inhibited colon cancer cells’ viability (HCT 116, 12% and 17%; SW-620, 13% and 19%, respectively). The highest firmness, cohesiveness, and viscosity index values, and the highest scores for color and taste, were obtained for yogurt with 3% APF, indicating that this is the optimal APF amount for the production of novel yogurt with functional properties.

Branislava Drašković, Tomislav Nikolić, Saša Jacović, D. Petrovic

Abstract Acute kidney damage associated with pregnancy occurs in 1/20.000 pregnancies. In developing countries, the main cause of the development of acute kidney damage is septic abortion, and preeclampsia in the developed countries of the world. Preeclampsia is defined as newly developed hypertension, proteinuria and swelling in pregnant women after the 20th week of gestation. It occurs due to disorders in the development of placenta and systemic disorders of the function of the endothelium of the mother. It is treated with methyldopa, magnesium sulfate and timely delivery. Urgent delivery is indicated if the age of gestation is ≥ 34 weeks. HELLP syndrome is a difficult form of preeclampsia. Its main characteristics are decreased platelet count, microangiopathic hemolysis anemia, increased concentration of aminotransferase in the serum and acute kidney damage. Severe HELLP syndrome is treated with emergency delivery, antihypertensives, magnesium sulfate, and in some cases plasmapheresis and hemodialysis. Acute fatty liver in pregnancy occurs because of decreased activity of the LCHAD enzyme of the fetus. Due to the reduced beta oxidation of fatty acids in the hepatocytes of the fetus, long chain fatty acids that cause damage to the mother’s hepatocytes are released. Swansea criteria are used for diagnosis, and the difficult form of the disease is treated with plasmapheresis and extracorporeal liver support. Atypical HUS is due to a reduced protein activity that regulates the activity of the alternative pathway of the complement system. Its main features are thrombocytopenia, microangiopathic hemolytic anemia and acute kidney damage. It is treated with plasmapheresis, and in case of resistance with eculizumab. Thrombotic thrombocytopenic purpura is due to decreased activity of the ADAMTS13 enzyme. It is characterized by thrombocytopenia, microangiopathic hemolytic anemia, high temperature, nervous system disorders and acute kidney damage. It is treated with plasmapheresis, and severe form of disease with corticosteroids and azathioprine. Early detection and timely treatment of acute kidney damage provides a good outcome for the mother and fetus.

Introduction: It is an undeniable fact that antidepressants can cause side effects. Antidepressants generally have a similar effect but they differ in their application safety, as well as their side effects. Aim: To determine differences in the frequency and intensity of antidepressant induced side effects in patients treated in primary care. Methods: The research was designed as a prospective, cross-sectional study, conducted on a voluntary and anonymous basis, and it included depression patients treated with antidepressant medications during 2013-2015 in Zenica-Doboj Canton using the Hamilton Depression Rating Scale and Toronto Side Effects Scale. Results: The total sample included 508 subjects. As a significant problem, abdominal pain was felt by 14% of subjects, indigestion by 19% of subjects, nausea by 15% of subjects, diarrhea by 9% of subjects, and constipation by 11% of subjects. 29% of subjects suffered from sweating, 20% suffered from a sudden heat stroke, 10% suffered from swelling, and 23% of them reported suffering from dry mouth as a significant problem. The prevalence of side effects in relation to how do they affect life and daily activities of subjects is statistically significant (P <0.000). Statistically significant side effects of SSRI antidepressants correlate with the duration of our subject’s treatment: perception of increased sleep (0.039) as well as decreased sleep (P = 0.009), sweating (P <0.001), sudden heat stroke (P <0.001), being without orgasm (P = 0.004), decreased libido (P <0.001), weight loss (P = 0.045). Conclusion: It is necessary to educate the patients about the nature and features of the depressive disorder, and to notify the patients of the expected course of recovery, as well as the need to adhere to the recommended therapy and the possible side effects of the medication.

Mahira Jahić, L. Kamerić, A. Hadžimehmedović

Introduction: Infection with human papillomavirus is the main cause of cervical carcinoma. In Bosnia and Herzegovina (BIH) 556 cases of cervical carcinoma are diagnosed annually, and 141 women die from it. Aim: To determine the most common HPV type found in ASCUS and LSIL and progression, regression and persistence of lesions. Methods: In a retrospective study, 11 051 PAP tests, performed in several private gynecological practices located in Tuzla Canton from January 2016 to December 2019, were analyzed. In processing of data , X2 – statistical method was used. Results: 11051 PAP test were inspected. Normal findings were found in 90.48% (N-10002) and pathological findings in 9.49% (N- 049). ASCUS was present in 4.9% (N-544), LSIL in 3.04% (N-337), HSIL in 0.74% (N-84), ASC-H in 0.27% (N-30) and AGC in 0.49% (N-55) of cases. The most common is HPV 16, found in 50.5% (N-44) of ASCUS and LSIL. Monoinfection with HPV 16 was found in 40.9% (N-18) ASCUS (N-3) and LSIL (N-15), and a combination of HPV 16 with other types like HPV 18, 31, 33, 39 in 59% (N-26). Progression of ASCUS lesion in HSIL 1.6% (N-2), and LSIL 9.6% (N-12). Progression of LSIL to HSIL was found in 9.0% (N-10). HSIL progresses significantly more frequent from LSIL (p<0.05) than from ASCUS changes. Lesions that progress into higher grade HSIL are HPV 16 positive. Progression into HSIL is not found in patients with low-risk HPV 6 and 11 infection. Conclusion: Women infected with HPV 16 have more a frequent progression of a lesion into higher grade HSIL. They should be intensively monitored because of the increased risk for development of cervical carcinoma.

S. Rosati, P. Franco, C. Fiandra, F. Arcadipane, P. Silvetti, E. Gallio, J. Panić, U. Ricardi et al.

One of the main problems during in the treatment of anal cancer with chemotherapy and radiation is the occurrence of Hematologic Toxicity (HT). In particular, during radiotherapy it is crucial to spare Bone Marrow (BM), since the radiation dose received by BM in pelvic bones predicts the onset of HT. In this direction, the most popular strategies are based on the identification of the hematopoietically active BM (actBM), that is the part of BM in charge of blood cells generation, using MRI, SPECT or PET, but no approached have been proposed based on CT. In this study we compare four different classifiers in recognizing actBM from CT images using 36 radiomic features. We used Genetic Algorithms (GAs) to simultaneously optimize the feature subsets and the classifier parameters, separately for three pelvic subregions: iliac bone marrow (IBM), lower pelvis bone marrow (LPBM), and lumbosacral bone marrow (LSBM). The obtained classifiers were applied to CT sequences of a cohort of 25 patients affected by carcinoma of the anal canal. Classifiers results were compared with the actBM identified from 18FDG-PET (reference standard, RS). It emerged that the performances of the 4 classifiers are similar and they are satisfactory for IBM and LSBM subregions (Dice > 0.7) whereas they are poor for LPBM (Dice < 0.5).

V. Giannini, Arianna Defeudis, S. Rosati, G. Cappello, L. Vassallo, S. Mazzetti, J. Panić, D. Regge et al.

Predicting response to neo-adjuvant chemotherapy of liver metastases (mts) using CT images is of key importance to provide personalized treatments. However, manual segmentation of mts should be avoid to develop methods that could be integrated into the clinical practice. The aim of this study is to evaluate if and how much automatic segmentation can affect a radiomics-based method to predict response to neoadjuvant chemotherapy of individual liver mts. To this scope, we developed an automatic deep learning method to segment liver mts, based on the U-net architecture, and we compared the classification results of a classifier fed with manual and automatic masks. In the validation set composed of 39 liver mts, the automatic deeplearning algorithm was able to detect 82% of mts, with a median precision of 67%. Using manual and automatic masks, we obtained the same classification in 19/32 mts. In case of mts with largest diameter > 20 mm, the precision of the segmentation does not impact the classification results and we obtained the same classification with both masks. Conversely, with smaller mts, we showed that a Dice coefficient of at least 0.5 should be obtained to extract the same information from the two segmentations. This are very important results in the perspective of using radiomics-based approach to predict response to therapy into clinical practice. Indeed, either precisely manually segment all lesions or refine them after automatic segmentation is a time-consuming task that cannot be performed on a daily basis.

Introduction: Calciphylaxis is a rare, but serious, kidney complication. Calciphylaxia is a vasculopathy of small blood vessels characterized by the deposition of calcium deposits in intimal arterioles with the consequent proliferation of intima, fibrosis and thrombosis. Aim: The aim was to show the significance of recognition of calciphylaxis relies on heightened clinical awareness of the presence of atypical skin nodules or ulcers that occur in patients with hemodialysis dependence and to characterize features of calciphylaxis or components of treatment that may lead to improved outcome. Case report: We present the case of 84-year-old woman with chronic kidney disease and diabetes mellitus as well as severely painful, firm, indurated plaques on the lower extremities. The plaques progressed to involve larger areas with associated local ulceration and necrosis. Laboratory testing revealed hyperparathyroidism and incisional skin biopsy confirmed calciphylaxis. Wound microbiology confirmed Staphylococcus aureus. Conclusion: The diagnosis can be based on clinical grounds, supported by histological analysis if possible. The laboratory workout must cover all the possible implications of chronic kidney disease with special attention to Ca+ and P+ values and evidence of skin or systemic infection. Calciphylaxis must be known by dermatologist as early diagnosis and proper management can be decisive for better prognosis.

The occurance of mid- and longterm uremic complications is related to the low clearance rate of middle and large molecule uremic toxins when hemodialysis (HD) alone is adopted. As the uremic toxins and their corresponding biological effects become increasingly clear, blood purification treatment that aims to remove these toxins, has developed from a stage of life-sustaining to improving the quality of life. The objective of this study was to evaluated demographic, clinical and laboratory data in patients who underwent the combination of maintenance hemodialysis with hemoperfusion (HP) and in those who recieved HD alone and to investigate whether this combination could improve the clearance rate of middle and large molecule uremic toxins. A total of 26 patients, who underwent routine hemodialysis, were assessed in this study. Those patients were randomly divided into three groups: Group 1 (7 patients) received combined treatment of HD with HP biweekly (HD 2 times a week with HD+HP once a week), whereas Group 2 (10 patients) was given HD with high flux dialyzer and Group 3 (9 patients) was given HD with low flux dialyzer 3 times a week. This study was followed for 4 months. Before and after the observational period demographic and clinical data were taken from the medical history and blood samples were taken for hemoglobin (Hb), iron (Fe), total iron binding capacity (TIBC), albumin (Alb), calcium (Ca), phosphorus (P04) and parathyroid hormone (PTH). This study included 13 female and 13 male patients with a mean age of 41, 62 + 11.12 and a mean dialysis duration of 62, 78+53, 33 months. When it comes to baseline characteristics, patients of the group 3 were significantly older than patients in other groups (p=0.001). At the end of the four months observation period, the same difference according to age was noticed (p=0.01). Also, HD+HP group had significantly higher values of TIBC (p=0.006) and significantly lower serum levels of P04 (p=0.001). EPO doses were very similar in group 1 and 2, but in group 3 there were noticeably lower than in those two groups but without a significant difference. The serum levels of albumin were higher in group 3 compared to the other two groups but also without statistical difference. No statistical difference between groups after the follow up period was observed in terms of Hb, Fe, PTH, Ca, BMI, duration of dialysis treatment and vascular access. When groups are viewed individually, in the HD+HP group serum P04 levels were significantly lower after the 4 months off the follow up period than it was at the beginning (p=0.031) and also TIBC was significantly higher (p=0.018). In group 2 the values of TIBC were significantly lower after the follow up period than it was at the beginning (p=0.025). No significant difference was noticed in group 3 but serum PTH levels tends to decrease after 4 months compared to baseline measurement. This combination treatment of HD with HP was superior to HD in reducing levels of phosphorus. These findings suggests a potential role of reducing the risk of cardiovascular events in this population especially when it is known that hyperphosphatemia has been pointed out as the primary culprit in the process of cardiovascular calcification. Also, patients who underwent the combined treatment showed higher values of TIBC but unfortunately no difference was noticed between Hb levels and EPO doses. These results eventually demonstrates their role in the improvement of renal disease anemia, which opens up the possibility of further research on a larger sample and over a longer period of time.

S. Hodžić, Hana Paleka

Abstract In a dynamic market, the city has become a main source of competitiveness, along with financial and economic benefits. Due to the processes of digitalization, a new concept has been developed, namely smart cities. This concept delivers economic and financial potential, not only to cities, but also to urban and local economic development. Therefore, to ensure the establishment of this concept, local government units, i.e., cities, need to have enough financial resources. In addition, the fiscal capacity of their local budgets should be sufficient. The objective of this paper is to evaluate the fiscal capacity of large cities in Croatia over the 2016-2018 period, as well as to present the financial support through funding schemes for the establishment of smart cities. After calculating the level of fiscal capacity of large cities, the analysis revealed interesting results. Only four large cities (Split, Rijeka, Zadar and Pula) achieved positive fiscal capacity in the observed period. This provides evidence of fiscal performance and fiscal capacity for the establishment of smart cities. This concept will enhance the quality of life, stimulate economic growth, sustain local government budgets and create new value for both investors and the local population.

P. Marešová, M. Prochazka, Sabina Baraković, Jasmina Baraković Husić, K. Kuča

A lack of nurses in the Czech Republic is an issue that has been under discussion for several years. The aim of this paper is to analyze the lack and need of general nurses and midwives in the Hradec Kralove region where the shortage is higher than the national average. The used methods are quantitative research and structured interviews, to determine the number of nurses in healthcare institutions. The study uses data obtained from publicly available sources, i.e., Czech Statistical Office (CSO) and the National Institute of Education (NIE). The shortage of nurses in the Hradec Kralove region can be expected by 2030 to be in the range between 647.6 and 667.1 nurses while maintaining the existing conditions, that is, five times more than at present. In addition to the commonly considered measures that appear in the country’s strategies—such as improving the quality of conditions during studies and during employment, specifying or adjusting the role and competency of nurses and midwives in the healthcare system, or unifying employment standards— a focus on promoting the nursing profession can be recommended. Schools and ministries should be encouraged to focus on and invest in the promotion of this profession, so as to play a key role in recruiting new students for the nursing field of study at a time when the nursing profession is perceived positively, as an embodiment of solidarity and selflessness.

B. Stanetic, M. Ostojić, T. Kovacevic-Preradovic, L. Kos, Kosana Stanetić, A. Nikolic, M. Bojić, K. Huber

Introduction Results of currently available trials have shown divergent outcomes in diabetic patients undergoing percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). Current guidelines do not recommend PCI in patients with diabetes and a SYNTAX score ≥ 23. Aim To compare all-cause 4-year mortality after revascularization for complex coronary artery disease (CAD) in diabetics. Material and methods The study group comprised consecutive patients with three-vessel CAD and/or unprotected left main CAD (≥ 50% diameter stenosis) without major hemodynamic instability, who were treated in two institutions with PCI or referred for CABG. Results Out of 342 diabetics, 177 patients underwent PCI and 165 patients were referred for CABG. The incidence of all-cause death was different between diabetics treated with PCI or CABG at 4 years (16/177, 9.0% vs. 26/165, 15.8%, respectively, p = 0.03). The difference was not evident in non-diabetics (PCI: 41/450, 9.1% vs. CABG: 19/249, 7.6%, p = 0.173). In diabetics, there was a higher incidence of all-cause mortality in PCI patients with intermediate-high (≥ 23) SYNTAX scores compared with those with low (0–22) SYNTAX scores (10/56, 17.9% vs. 6/121, 5.0%, respectively, p < 0.01). On the other hand, diabetics who underwent CABG showed similar mortality rates irrespective of the SYNTAX scores (SYNTAX 0–22: 3/29, 10.3%; SYNTAX ≥ 23: 23/136, 11.9%, p = 0.46). In the subgroup analysis, there was no interaction according to presence or absence of left main CAD (p for interaction = 0.12) as well as according to diabetes status (p for interaction = 0.38), whereas gender and SYNTAX scores were differentiators between PCI and CABG with a p for interaction < 0.1. Conclusions Our analysis supports recent evidence that diabetes is not a differentiator between PCI and CABG.

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