Regional differences in expression of molecular markers during formation of the expanded subplate zone in the human fetal cerebral cortex

U doktorskoj disertaciji prikazane su regionalne citoarhitektonske razlike između čeone, tjemene, zatiljne i cingularne moždane kore u fazi ekspanzije i stvaranja subplate zone (od 13. do 15. TNZ). Pomoću imunohistokemijskih metoda na prenatalnom postmortalnom tkivu mozga čovjeka praćena je dinamika ekspresije molekularnih biljega. Rezultati su pokazali kako regionalne razlike između pojedinih izokortikalnih regija fetalne moždane kore čovjeka postaju vidljive rano tijekom prenatalnog razvoja, a postaju najistaknutije upravo tijekom razdoblja formiranja SP-a. Proces stvaranja SP-a iz dubokog dijela KP prikazan je biljegom SP neurona TBR1. Pokazano je i kako je obrazac formiranja SP-a važan kriterij za diferenciranje dorzalne izokortikalne i ventralne mezokortikalne cingularne moždane kore u ranom fetalnom razdoblju. Nadalje, karakteristike rane diferencijacije mezokortikalne cingularne moždane kore su: proširenje MZ, suženje KP i SVZ. Jedna od glavnih karakteristika mezokortikalnog dijela cingularne moždane kore je nepotpuna ekspanzija SP-a, a navedeno je prikazano pomoću biljega projekcijskih neurona. Analiza ranog razvoja cingularne moždane kore važna je radi njezine uloge u stvaranju ranih neuralnih krugova uključenih u ponašanje i emocije. Nadalje, poznavanje regionalnog razvoja bitno je za razumijevanje arealne diferencijacije i kasnije funkcionalne specifikacije moždane kore što je preduvjet za razumijevanje neurorazvojnih poremećaja.In the doctoral thesis, regional cytoarchitectonic differences between the frontal, parietal, occipital, and cingulate cortex were analyzed in the the subplate formation phase (13 to 15 PCW). Immunohistochemical methods were used on prenatal postmortem human brain tissue to analyze the molecular markers` expression pattern dynamics. The results showed that regional differences between isocortical regions of the human fetal cortex become visible early during prenatal development, and are most prominent during the SP formation period. The SP formation process is shown with the SP neuron marker TBR1. Additionally, we showed that the SP formation pattern is an important criterion for differentiating the dorsal isocortical and ventral mesocortical cingulate cortex in the early fetal period. Furthermore, the early mesocortical cingulate cortex is characterized by the widening of the MZ and the narrowing of the CP and SVZ. One of the main characteristics of the mesocortical part of the cingulate cortex is an incomplete SP expansion, and this is shown in the results using projection neuron markers. The early cingulate cortex development analysis is important because of its involvement in the formation of early neural circuits involved in behavior and emotions. Furthermore, analysis of regional differences is essential for understanding areal differentiation and later functional specification of the cerebral cortex, which is a prerequisite for understanding diverse neurodevelopmental disorders

Recurrence of hepatocellular carcinoma after liver transplantation

Transplantacija jetre predstavlja kurativnu opciju za hepatocelularni karcinom (HCC), no recidiv bolesti i dalje se javlja u 8–20 % bolesnika, osobito u uvjetima trajne imunosupresije. Glavni rizični čimbenici uključuju mikrovaskularnu i makrovaskularnu invaziju, visoko tumorsko opterećenje, povišeni alfa-fetoprotein, transplantaciju izvan prihvaćenih kriterija i nedovoljan odgovor na prethodnu terapiju. Novija istraživanja ukazuju i na potencijalnu ulogu metaboličkih poremećaja u nastanku recidiva. U ovom diplomskom radu provedeno je retrospektivno istraživanje na 231 bolesniku transplantiranom zbog HCC-a u KB Merkur (2016.–2024.). Analizirani su klinički, laboratorijski i patohistološki podaci, uključujući primjenu TACE terapije i razine takrolimusa. Recidiv je zabilježen kod 10,39 % bolesnika. Statistički značajna povezanost s recidivom utvrđena je za veći broj tumorskih čvorova, prisutnost mikrovaskularne i limfovaskularne invazije te viši stadij bolesti. Također, zabilježena je veća učestalost hiperlipoproteinemije u skupini s recidivom, što može upućivati na metaboličku komponentu rizika. Zaključno, broj tumorskih čvorova i mikrovaskularna invazija potvrđeni su kao ključni čimbenici rizika za recidiv HCC-a, dok metabolički poremećaji mogu dodatno doprinijeti procjeni rizika. Potrebna su daljnja prospektivna istraživanja za precizniju stratifikaciju bolesnika nakon transplantacije.Liver transplantation represents a curative treatment option for hepatocellular carcinoma (HCC), but disease recurrence still occurs in 8–20% of patients, particularly in the context of lifelong immunosuppression. Major risk factors include microvascular and macrovascular invasion, high tumor burden, elevated alpha-fetoprotein levels, transplantation beyond accepted criteria, and inadequate response to downstaging therapies. Recent studies also suggest a potential role of metabolic disorders in increasing the risk of recurrence. This thesis presents a retrospective study of 231 patients who underwent liver transplantation for HCC at Merkur University Hospital between 2016 and 2024. Clinical, laboratory, and histopathological data were analyzed, including the use of TACE therapy and tacrolimus levels. Disease recurrence was observed in 10.39% of patients. Statistically significant associations with recurrence were found for a higher number of tumor nodules, the presence of microvascular and lymphovascular invasion, and more advanced tumor stage. Additionally, hyperlipoproteinemia was more frequent in the recurrence group, indicating a possible metabolic contribution to recurrence risk. In conclusion, the number of tumor nodules and microvascular invasion were confirmed as key risk factors for HCC recurrence, while metabolic factors such as hyperlipoproteinemia may have additional prognostic value. Further prospective studies are needed to improve risk stratification after liver transplantation

Comparable outcomes after busulfan- or treosulfan-based conditioning for allo-HSCT in children with ALL: results of FORUM

The superiority of total body irradiation (TBI)-based vs chemotherapy conditioning for allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with acute lymphoblastic leukemia (ALL) has been established in the international, prospective phase-3 FORUM study, randomizing 417 patients aged 4-18 years in complete remission (CR), who received allo-HSCT from HLA-matched sibling or unrelated donors. Because of the unavailability of TBI in some regions and to accommodate individual contraindications, this study reports the prespecified comparison of outcomes of patients receiving busulfan (BU)- or treosulfan (TREO)-based regimens from 2013 to 2018. Overall, 180 and 128 patients received BU/thiotepa (THIO)/fludarabine (FLU) or TREO/THIO/FLU, respectively. Data were analyzed as of February 2023, with a median follow-up of 4.2 years (range, 0.3-9.1). 3-year overall survival was 0.71 (BU, 95% confidence interval [0.64-0.77]) and 0.72 (TREO, [0.63-0.79]) and 3-year event-free survival was 0.60 (BU, [0.53-0.67]) and 0.55 (TREO, [0.46-0.63]). The 3-year cumulative incidence of relapse (BU, 0.31 [0.25-0.38]; TREO, 0.36 [0.27-0.44]); and nonrelapse mortality (BU, 0.08 [0.05-0.13]; TREO, 0.09 [0.05-0.15]) were comparable. One case of fatal veno-occlusive disease occurred in each group. No significant differences in acute and chronic graft-versus-host disease (GVHD) or 3-year GVHD-free and relapse-free survival (BU, 0.48 [0.41-0.55]; TREO, 0.45 [0.37-0.54]) were recorded. Outcomes for patients in first and second CR were similar irrespective of the regimen. In conclusion, BU/THIO/FLU or TREO/THIO/FLU regimens can be an alternative to TBI for patients with ALL aged >4 years with contraindications or lack of access to TB

Quiescent cells maintain active degradation-mediated protein quality control requiring proteasome, autophagy, and nucleus-vacuole junctions

Many cells spend a major part of their life in quiescence, a reversible state characterized by a distinct cellular organization and metabolism. In glucose-depleted quiescent yeast cells, there is a metabolic shift from glycolysis to mitochondrial respiration, and a large fraction of proteasomes are reorganized into cytoplasmic granules containing disassembled particles. Given these changes, the operation of protein quality control (PQC) in quiescent cells, in particular the reliance on degradation-mediated PQC and the specific pathways involved, remains unclear. By examining model misfolded proteins expressed in glucose-depleted quiescent yeast cells, we found that misfolded proteins are targeted for selective degradation requiring functional 26S proteasomes. This indicates that a significant pool of proteasomes remains active in degrading quality control substrates. Misfolded proteins were degraded in a manner dependent on the E3 ubiquitin ligases Ubr1 and San1, with Ubr1 playing a dominant role. In contrast to exponentially growing cells, the efficient clearance of certain misfolded proteins additionally required intact nucleus-vacuole junctions (NVJ) and Cue5-independent selective autophagy. Our findings suggest that proteasome activity, autophagy, and NVJ-dependent degradation operate in parallel. Together, the data demonstrate that quiescent cells maintain active PQC that relies primarily on selective protein degradation. The necessity of multiple degradation pathways for the removal of misfolded proteins during quiescence underscores the importance of misfolded protein clearance in this cellular state

European general practitioners’ attitudes towards person-centred care and factors that influence its implementation in everyday practice: The protocol of the cross-sectional PACE GP/FP study in 24 European countries

Background: Person-centred care (PCC) is a fundamental principle in general practice, emphasising practices tailored to individual patient preferences, needs, and values. Despite the importance of PCC, general practitioners (GPs) face obstacles in effectively implementing it, with associated factors remaining unclear. Objectives: The PACE GP/FP study aims to explore GPs' attitudes towards PCC and the factors facilitating or hindering its implementation in daily practice across European countries. This paper outlines the PACE GP/FP study protocol. Methods: The cross-sectional design with data collection via an online survey distribution to GPs in 24 European countries. Study instruments include two validated questionnaires (Perceived Stress Scale (PSS) and Patient Physician Orientation Scale (PPOS)) and additional items covering general information about the doctor and their practice, as well as facilitators and barriers to PCC. These additional items were specifically developed for the study, translated using the forward-backward method, evaluated through cognitive debriefing, and integrated into the REDCap platform to create language and country-specific survey links. The STROBE checklist guides the reporting of the manuscript. Conclusion: The PACE GP/FP study will provide a comprehensive exploration of GPs' attitudes towards PCC and the factors shaping its practice in Europe. The findings from the PACE GP/FP study will provide evidence for designing future implementation strategies and guide targeted interventions to promote PCC in primary care across Europe

Signs of arterial and respiratory function impairment in children with inflammatory bowel disease

Uvod: Cilj ovog istraživanja je ispitati promjene na arterijama i dišnom sustavu djece s upalnom bolečću crijeva (IBD), njihovu međusobnu povezanost te povezanost s upalnim parametrima, trajanjem i aktivnošću bolesti. Ispitanici i metode: Radi se o presječnom istraživanju parova, a uključeno je 55-tero bolesnika s novootkrivenom bolešću, 53-je u remisiji te 53-je zdravih ispitanika. Debljina intime-medije karotidnih arterija mjerena je ultrazvučno, a ostali arterijski parametri oscilometrijski. Plućna funkcija procijenjena je spirometrijom, a frakcija izdahnutog dušičnog oksida (FeNO) kemiluminiscentnim analizatorom. Rezultati: Nismo uočili statistički značajnu razliku među skupinama u brzini pulsnog vala kroz aortu (PWVao)(P=0.515), markeru krutosti arterija. Bolesnici s IBD-om imaju značajno niži augmentacijski indeks (Aix)( P<0.001) te višu srčanu frekvenciju (HR)(P=0.001), što korelira s trajanjem bolesti (ρ(159)=0.22, P=0.006). Nismo uočili značajnu razliku u parametrima spirometrije, dok je FeNO parametar viši kod aktivne bolesti (P=0.025). Zaključak: Kod djece s IBD-om još nema strukturnih promjena na arterijama, no izmijenjeni oblik pulsnog vala te povišen HR parametar, koji korelira s trajanjem bolesti, reflektiraju rane funkcionalne promjene. Kod aktivne bolesti upalom je zahvaćen i dišni sustav, premda je respiratorna funkcija, mjerena spirometrijom, još uvijek normalna.Introduction: The aim was to examine the arterial and respiratory changes in children with inflammatory bowel disease (IBD), their interrelationship and the relationship with inflammatory parameters, disease duration and activity. Subjects and methods: This cross-sectional case-control study included 55 patients with the newly diagnosed disease, 53 in remission and 53 healthy subjects. The carotid intima-media thickness was measured by ultrasound, and the other arterial parameters were measured by oscillometry. Lung function was assessed by spirometry, and fraction of exhaled nitric oxide (FeNO) by chemiluminescence analyser. Results: There is no statistically significant difference in the pulse wave velocity (PWVao)(P=0.515), an arterial stiffness marker. IBD patients have a lower augmentation index (Aix)(P<0.001). The heart rate correlates with the disease duration (ρ(159)=0.22, P=0.006). Spirometry is normal, while FeNO is higher in active disease (P=0.025). Conclusion: In children with IBD, the arteries have no structural changes. However, the altered pulse wave and the heart rate that increases with the disease duration, reflect early functional changes. In active disease, inflammation also affects the respiratory system, although respiratory function measured by spirometry is still normal

Treatment of patients with ovarian cancer after neoadjuvant chemotherapy

Zbog nedostatka dovoljno pouzdanih probirnih metoda i učestalog postavljanja dijagnoze u kasnom stadiju, karcinom jajnika predstavlja veliki dijagnostički izazov. Simptomi bolesti često su nespecifični – bolesnice često pripisuju simptome stresu, promjenama u prehrani ili poremećajima peristaltike, što u većini slučajeva rezultira postavljanjem dijagnoze u uznapredovalom stadiju bolesti, najčešće stadiju III ili IV. Povijesno gledano, primarna citoredukcija (PDS, engl. primary debulking surgery) je bio temelj liječenja karcinoma jajnika, s ciljem maksimalnog smanjenja tumorske mase prije početka kemoterapije. Citoredukcija ima osim terapijske i dijagnostičku vrijednost jer omogućuje procjenu opsega proširenosti bolesti i mogućnosti potpune resekcije. Optimalna citoredukcija, definirana kao rezidualna bolest karcinoma jajnika jednaka ili manja od 1 cm, je najvažniji prognostički čimbenik. Manji rezidualni tumori ili kompletna citoredukcija povezani su s duljim preživljenjem i boljim terapijskim ishodom. Sve više studija danas zagovara primjenu neoadjuvantne kemoterapije (NACT, engl. Neoadjuvant chemotherapy) kao terapijsku opciju kod bolesnica koje zbog lošeg općeg stanja, opsežne tumorske mase ili komorbiditeta nevezanih uz bolest nisu kandidati za primarni kirurški zahvat. Uvođenje NACT u takvih bolesnica je poboljšalo stopu optimalne citoredukcije i smanjilo učestalost postoperativnih komplikacija te ujedno poboljšao prognozu i preživljenje. Odluka između PDS i NACT mora biti individualizirana, uzimajući u obzir biologiju tumora, funkcionalni status i komorbiditete bolesnice i mogućnost optimalne resekcije.Because of a lack of effective screening methods and diagnosis often in advanced stages, ovarian cancer is a diagnostic challenge. Symptoms are often unspecific –usually written off to stress, dietary changes or digestive disturbances. That results in diagnosis in stages III or IV. Historically speaking, primary debulking surgery (PDS) was the foundation of ovarian cancer treatment, aiming for maximal tumor resection before chemotherapy. Cytoreduction has not only a therapeutic value, but also a diagnostic one, for it assesses the likelihood of achieving a complete resection. Optimal cytoreduction, defined as residual disease equal to or less than one centimeter, is the most important prognostic factor. A complete resection or smaller residual tumors are connected with prolonged survival and better outcome. More and more studies advocate for neoadjuvant chemotherapy (NACT) as an alternative approach for patients with low performance status, large tumor masses and comorbidities that enable them to receive PDS. The emergence of NACT has enhanced the ability to achieve optimal cytoreduction and has reduced the rate of postoperative complications It has also improved overall survival and outcomes in those patients, which proves it´s noninferiority to PDS. The choice between the two approaches must be made on an individual level, taking tumor biology, patient´s performance status and comorbidities into account, while also assessing the possibility of optimal cytoreduction

Each Indicator of Socioeconomic Status (Education, Occupation, Income, and Household Size) Is Differently Associated with Children’s Diets: Results from a Cross-Sectional CroCOSI Study

Background: There has yet to be an agreement on which specific socioeconomic status (SES) indicator most effectively reflects disparities in children’s diets. However, children from lower SES backgrounds are particularly vulnerable, as research in other countries indicates that their diets contain fewer fruits and vegetables and more sweetened beverages. This paper aims to evaluate the associations between dietary habits and various SES indicators (education, occupation, income, and household size) among a representative sample of children in Croatia aged 7–10. Methods: Parents of children were asked to complete a questionnaire that contained indicators of their children’s dietary habits and socioeconomic status (n = 5608). Associations between SES and children’s dietary habits were assessed using logistic regression models. Results: The mother and father’s educational attainment were strongly positively associated with breakfast consumption. Children of parents with a lower educational level consumed sweetened beverages, sweet snacks, and fast food slightly more often than children in families with a higher educational background. The mother’s education was inversely associated with vegetable and cereal consumption, while the father’s education was inversely associated with fruit and bakery product consumption. Meanwhile, household income per unit had a significant influence on the consumption of soft drinks and bakery products. Household size had a significant influence solely on sweet snack consumption. Conclusions: Each SES indicator showed an independent association with at least one particular dietary habit, except for the parent’s employment status

Small intestinal transplantation

Transplantacija tankog crijeva je složeni kirurški zahvat namijenjen pacijentima s kroničnim zatajenjem crijeva nakon iscrpljenih svih drugih terapijskih mogućnosti. Najčešći uzrok zatajenja je sindrom kratkog crijeva, a transplantacija se razmatra kad dugotrajna parenteralna prehrana dovede do ozbiljnih komplikacija poput zatajenja jetre, tromboza ili infekcija. Indikacije i kontraindikacije su strogo definirane, a odluku o transplantaciji donosi multidisciplinarni tim. Transplantacija tankog crijeva može biti izolirana, simultana transplantacija jetre i tankog crijeva te multivisceralna. Davatelji su najčešće preminule osobe, a glavni koraci operacije su uklanjanje primateljevog crijeva, stvaranje krvožilnih i crijevnih anastomoza te postavljanje privremene ileostome radi olakšanog postoperativnog praćenja funkcije presatka. Imunosupresivna terapija neophodna je za sprječavanje odbacivanja, ali povećava rizik od infekcija i drugih komplikacija. Odbacivanje presatka i infekcije najčešće su komplikacije, osobito u prvim mjesecima nakon zahvata. Praćenje uključuje redovite endoskopske preglede i biopsije, dok se biomarkeri i slikovne metode koriste kao dopunski alati. Postoperativna skrb naglašava što ranije uvođenje enteralne prehrane, uz postupno ukidanje parenteralne prehrane. Pacijenti se dugoročno prate zbog mogućih metaboličkih i onkoloških komplikacija. Infekcije i bolest presatka protiv primatelja vodeći su uzroci smrtnosti i zahtijevaju brzo prepoznavanje i liječenje. U Hrvatskoj se transplantacija tankog crijeva izvodi samo u Kliničkoj bolnici Merkur, a broj zahvata je malen zbog rijetkih indikacija. Transplantacija tankog crijeva danas je određenim pacijentima jedina šansa za preživljenje i poboljšanje kvalitete života, ali zahtijeva stručan i multidisciplinaran pristup te dugotrajno praćenje zbog visokog rizika od komplikacija.Small intestinal transplantation is a complex surgical procedure performed in patients with chronic intestinal failure when all other therapeutic options have been exhausted. The most common cause of intestinal failure is short bowel syndrome, typically resulting from extensive intestinal resections or congenital disorders. Transplantation is warranted when prolonged parenteral nutrition leads to life-threatening complications such as liver failure, venous thrombosis or recurrent infections. Indications and contraindications are strictly defined, and the decision to proceed with transplantation is made by a multidisciplinary team. Types of intestinal transplantation include isolated intestine, combined liver and intestine and multivisceral transplantation. Donors are usually deceased individuals, and the main steps of the procedure include removal of the recipient’s intestine, creation of vascular and intestinal anastomoses and placement of a temporary ileostomy for postoperative monitoring of graft function. Immunosuppressive therapy is necessary to prevent rejection, but it increases the risk of infections and other complications. Graft rejection and infections are the most common complications, especially in the first months after the procedure. Monitoring includes regular endoscopic evaluation and biopsies, while biomarkers and imaging methods are used as supplementary tools. Postoperative care emphasizes early introduction of enteral nutrition with gradual discontinuation of parenteral nutrition. Patients are monitored long-term for possible metabolic and oncological complications. Infections and graft-versus-host disease are the leading causes of mortality and require prompt recognition and treatment. In Croatia, intestinal transplantation is performed only at Clinical Hospital Merkur, and the number of procedures is small due to rare indications. Intestinal transplantation is the only chance for survival and improved quality of life for certain patients, but requires expert and multidisciplinary management and long-term monitoring due to the high risk of complications