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Introduction/Objectives: Traumatic brain injury (TBI) occurs as a consequence of a mechanical force to the skull and causes life-long changes in the central nervous system (CNS). Neurodegenerative sequelae of TBI commonly include proteinopathies, such as the one involving the TAR DNA binding protein 43 (TDP-43). TDP-43 proteinopathy results in both gain of function, because of the formation of cytosolic aggregates that inhibit normal cellular mechanisms, and loss of function, with the lack of TDP-43 physiological properties. Repetitive brain traumas, which are common after sport and violent injuries, have been recognized as a risk factor for amyotrophic lateral sclerosis (ALS). There is less evidence of the association of a single brain trauma with TDP-43 proteinopathy. Studies have shown that brain injury can induce changes in the parts of the CNS that are distant from the initial trauma site, e.g., the hippocampus and cerebellum. The aim of this study was to investigate pathophysiological changes, including the ones related to the TDP-43 proteinopathy, of the mice spinal cord after single moderate and repetitive mild TBI. Participants, Materials/Methods: Single moderate brain trauma was induced over the left parietal cortex by the lateral fluid injury (LFPI) apparatus. A separate cohort of mice received repetitive mild TBI (RT) using the weight drop method, twice a day for five days in a row. Animals were sacrificed 14 days after LFPI or the last mild brain trauma and their spinal cords were prepared for immunohistological analyses. For both trauma models, sham-injured mice were used as the control group. Results: Cresyl violet staining did not reveal any significant morphological changes in the spinal cords of both trauma groups compared to their matching controls. Also, differences in the spinal cord white matter were not detected for RT and LFPI trauma groups of mice at this time point. The significant mislocalization of spinal cord TDP-43 was observed after both single moderate and repetitive mild brain injury. Cytoplasmic TDP-43 staining was evident in neurons but not in astrocytes of tested trauma groups. Conclusions: Our preliminary results suggest that both single and repetitive brain trauma can induce subtle pathophysiological changes, such as TDP-43 mislocalization, in CNS regions distant from the initial site of trauma. This work was supported by the University of Rijeka, Croatia, project uniri-biomed-18-199 to K.P. and uniri- biomed-18-204 to P.D.

Pharmacogenomics can help optimize and personalize pharmaceutical treatments by improving drug efficacy and safety and potentially reducing treatment costs. Adjuvant fluoropyrimidine-based chemotherapy is a frequently used in a great number of neoplasms, especially of 5-fluorouracil (5-FU) and capecitabine (CAP) which are metabolized through dihydropyrimidine dehydrogenase (DPD) enzyme in the liver encoded by DPYD gene. Up to 2-8% of the population (depending on ethnicity) have DPYD variations leading to DPD deficiency and an increased risk of severe toxicity when treated with regular doses. Our aim was to investigate the cost-effectiveness of DPYD genotyping considering its impact on clinical outcome and healthcare costs. The analysis of the literature showed that DPYD genotyping in stage 3 colorectal cancer is a cost- effective strategy that improves survival by 0.0038 QALYs at an ICER of $20, 506/QALY. Probabilistic sensitivity analysis showed that DPYD genotyping was preferred to no screening in 96.2% of iterations, at a willingness-to-pay threshold of $50, 000/QALY. In one retrospective study of 31 patients showed that hospitalization costs related to toxicity of five patients who tested positive for DPYD mutations were €155, 083. The cost of prospectively testing all patients starting first-line 5FU-based therapy over the same period would have been €23, 718, representing a cost saving of €131, 365 through the avoidance of these admissions alone. In conclusion, DPYD genotyping presents a considerable cost-effective method prior to fluropyrimidine-based adjuvant chemotherapy. Adjustment of the treatment dose prevents potentially severe and fatal toxicities and is worth implementing in daily clinical practice.

Summary. The carbohydrate portion of proteins and lipids mediates a variety of biological processes. Revealing their underlying principles is a challenging task that could contribute to a better understanding of many patho/physiological conditions. On the other hand, the interest in extracellular vesicles (EVs) has increased in recent years due to their involvement in intercellular communication leading to an array of functional and structural changes in recipient cells. Their characterization uncovered an exceptional diversity in size, morphology, as well as in membrane and cargo content. Monitoring/analysis of surface glycosylation of EVs originating from the prostate, termed prostasomes, revealed their substantial contribution to the complexity of seminal plasma (SP) glycome. Heterogeneity of surface glycans confirm the existence of several prostasome subpopulations. Presentation of surface glycans on prostasomal membrane is strongly affected by co-localized membrane-associated glycoproteins and tetraspanins. They appear to be organized in established/regular distribution patterns on membrane domains. Surface glycans are a component of EVs membrane that affects its functionality and potentially a distinction marker of prostasome subpopulations. Further understanding of the complex composition of glycans on EVs might explain the relation of their structure with functional alterations in distinct patho/physiological conditions.

Kakovost zraka je eden izmed najpomebnejših dejavnikov notranjega okolja saj ima slaba kakovost številne negativne kratkotrajne ali dolgotrajne učinke na ljudi v prostoru. Ker pa ljudje v današnjem času preživimo skoraj 90 % časa v zaprtih prostorih in je zrak tam v večini primerov bolj onesnažen kot zunaj, je potrebno za ustrezno kakovost zagotoviti učinkovite ukrepe. V diplomskem delu sem naredila pregled standardov iz omenjenega področja, raziskala škodljive vplive glavnih onesnaževal na zdravje učencev in zaposlenih v dvanajstih osnovnih šolah, kjer so bile meritve izvedene in podala ukrepe za izboljšanje. Air quality is one of the most important factors of the indoor environment, as poor quality has high negative short-term or long-term effects on the people in the room. However, since people today spend almost 90% of their time indoors and the air there is in most cases more polluted than outside, it is necessary to provide effective measures for adequate quality. In my thesis, I made an overview of the standards in the mentioned field, investigated the harmful effects of the main pollutants on the health of students and employees in twelve elementary schools, where the measurements were carried out, and gave measures for improvement.

Ciljevi istraživanja: U modelu jedne umjerene traumatske ozljede mozga (engl. traumatic brain injury, TBI) odrediti pojavnost i opseg izražaja TDP-43 u ipsilateralnom i kontralateralnom korteksu i hipokampusu, usporediti TDP-43 subcelularni izražaj nakon jedne umjerene TBI i blage repetitivne TBI (engl. repetitive mild traumatic brain injury, rmTBI), te odrediti povezanost TDP-43 izražaja s biljezima sinaptogeneze i upale nakon TBI. Materijal i metode: Metodom lateralne ozljede tlakom tekućine (engl. lateral fluid percussion injury, LFPI) izazvana je jedna TBI umjerene jakosti u miša, dok je metodom pada utega izazvana rmTBI. Izražaj proteina od interesa analiziran je u citoplazmatskim i nuklearnim frakcijama ipsilateralnog i kontralateralnog korteksa i hipokampusa u životinja LFPI skupina, te frontalnom korteksu i hipokampusima rmTBI miševa. Histološkim tehnikama praćene su moždane promjene, a imunofluorescentnim tehnikama subcelularna lokalizacija TDP-43 te povezanost s biljezima upale i sinaptogeneze. Rezultati: Ipsilateralni korteks LFPI životinja najoštećenija je moždana struktura koju prati dezintegracija bijele tvari. U ipsilateralnom korteksu pronađena je translokacija TDP-43 u citoplazmu prvog i trećeg dana nakon LFPI u neuronima i mikrogliji. U ipsilateralnom hipokampusu uočen je pad izražaja nuklearnog TDP-43 trećeg dana od jedne umjerene TBI. Četrnaest dana nakon jedne umjerene LFPI, ali i rmTBI uočen je porast izražaja nuklearnog TDP-43 u ipsilateralnom, odnosno frontalnom korteksu. Detektiran je porast izražaja fragmentiranog TDP-35 te značajan porast izražaja fosforiliranih oblika P-TDP-43 i P-TDP-35 u ipsilateralnom hipokampusu četrnaest dana nakon jedne LFPI, ali ne i u hipokampusu rmTBI životinja. U ipsilateralnom korteksu LFPI životinja uočena je povezanost biljega upale NF-κB i TDP-43 trećeg dana u području uz najveće oštećenje te povećan izražaj biljega iNOS prvog dana uz povezanost s TDP-43 u području najvećeg oštećenja. Pad izražaja PSD-95 uz gubitak neurona ipsilateralnog korteksa praćen je pozitivnom korelacijom s TDP-43 prvog, trećeg i četrnaestog dana od LFPI. Porast izražaja proteina APP uočen je subakutno samo u rmTBI skupini životinja, a proteina P-tau samo u LFPI miševa. Zaključak: Jedna umjerena LFPI uzrokuje deregulaciju TDP-43 u ipsilateralnom korteksu i hipokampusu koja utječe na pojedine biljega upale i sinaptogeneze. Navedene promjene ovise o zadobivenoj traumi mozga i od potencijalne su važnosti u farmakoterapiji TBI., Objectives: To determine the incidence and extent of TDP-43 expression of the ipsilateral and contralateral cortex and hippocampus after a single moderate traumatic brain injury (TBI), compare TDP-43 subcellular localization after single moderate TBI and repetitive mild traumatic brain injury (rmTBI), and determine the association of TDP-43 expression with markers of synaptogenesis and inflammation after TBI. Material and methods: Lateral fluid percussion injury (LFPI) was used to induce the single moderate TBI, while the weight drop method was used to cause the rmTBI. The levels of expression of proteins of interest were analyzed in the cytoplasmic and nuclear fractions of the ipsilateral and contralateral cortex and hippocampus in the animals of the LFPI and in the frontal cortex and the hippocampi of rmTBI mice. Histological techniques were used to determine brain changes, and immunofluorescent techniques were used to monitor the subcellular localization of TDP-43 and its association with some markers of inflammation and synaptogenesis. Results: The ipsilateral cortex of the LFPI group is the most damaged brain structure accompanied by white matter disintegration. Also, a translocation of TDP-43 into the cytoplasm was detected on the first and third day after LFPI in the neurons and microglia of the ipsilateral cortex. In the ipsilateral hippocampus, a decrease in the expression of nuclear TDP-43 was found on the third day after one moderate TBI. Fourteen days after LFPI and rmTBI, an increase in nuclear TDP-43 expression was observed in the ipsilateral and frontal cortex, respectively. Significant increases in the expression of TDP-35, P-TDP-43 and P-TDP-35 were detected subacutely only in the ipsilateral hippocampus of the LFPI animals. The association with inflammation marker NF-κB and TDP-43 was found around the area of the greatest cortical damage. Also, significant iNOS expression with the interconnection with TDP-43 was evident in the ipsilateral cortex on the first day after LFPI. The decline in PSD-95 expression with loss of ipsilateral cortical neurons was followed by a positive correlation with TDP-43 on the first, third and fourteenth day after LFPI. An increase in APP protein expression was observed subacutely only in the rmTBI group of animals, and P-tau protein only in the LFPI mice. Conclusion: One moderate LFPI causes deregulation of TDP-43 in the ipsilateral cortex and hippocampus and it is related to some markers of inflammation and synaptogenesis. These changes depend on the type of the acquired brain trauma and are of potential importance in TBI pharmacotherapy.

Introduction: Traumatic brain injury (TBI) occurs as a consequence of mechanical injury to the brain and is considered to be the leading cause of death and disability in young individuals. Inflammation, one of the secondary injury pathophysiological processes, contributes to posttraumatic brain damage and lifelong cognition and memory impairments. Inducible nitric oxide synthase (iNOS) is a cytoplasmic enzyme that forms the signalling molecule nitric oxide involved in neuroinflammation. In TBI, it has been also shown that mislocalization of TAR DNA-binding protein 43 (TDP-43) contributes to post-injury pathology. We investigated acute changes in the iNOS expression following single moderate TBI in wild-type mice in different brain regions and its colocalization with TDP-43. Materials/Methods: Single moderate lateral fluid percussion injury was induced over the left parietal cortex of the male adult C57BL/J mice. Animals were sacrificed one or three days after TBI and their brains were prepared for Western blot or immunohistological analyses. Animals of the control group were sacrificed 1 day after the sham injury procedure. Results: An increase in the iNOS expression was observed in the ipsilateral cortex one day after the brain trauma. In the tested time points, changes in the iNOS expression were not detected in the ipsilateral hippocampus and contralateral cortex, and hippocampus of traumatized animals. Also, a colocalization of iNOS and TDP-43 in the cytoplasm was observed in the most damaged part of the ipsilateral cortex one day after TBI. Conclusions: Transient acute changes in the proinflammatory marker iNOS in the ipsilateral cortex are induced by single moderate brain trauma. Cytoplasmic colocalization of iNOS and TDP-43 in the most damaged part of the ipsilateral cortex suggests the interconnection of these proteins. This work was supported by the University of Rijeka, Croatia, project number uniri-biomed-18-199 to K.P.

Traumatic brain injury (TBI) is a lifelong condition that leads to progressive brain damage and cognitive disturbances. TBI has been recognized as a risk factor for the development of numerous neurodegenerative diseases, such as Alzheimer and Parkinson’s disease, amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Proteinopathy of TAR DNA binding protein 43 (TDP-43) is considered the most important pathological characteristic of ALS and FTLD, that is characterized by permanent translocation of TDP-43 in the cytoplasm where this protein undergoes hyperphosphorylation, ubiquitination and fragmentation with subsequent inclusion formation. Toxic effects of TDP-43 inclusions have been demonstrated in the neurons and microglia, but deficiency of nuclear TDP-43 has numerous cell consequences too. Although cellular processes that induce TDP-43 proteinopathy are still unclear, it has been proven that inflammation can induce these changes. Interconnection of TDP-43 with some synaptic proteins has also been found. TDP-43 inclusions have been detected in the brain tissue after repetitive trauma in numerous clinical and preclinical investigations and it is mostly considered that this type of proteinopathy is only a consequence of repetitive, but with a single TBI, results have thus far been inconclusive. The aim of this study was to investigate the changes in the TDP- 43 localization, fragmentation, and phosphorylation in different brain regions after a single moderate TBI in mice. Additionally, we examined the association of TDP-43 expression with some markers of synaptogenesis and inflammation. This work was supported by the University of Rijeka, Croatia, project-uniri- biomed-18-199 to K.P. and Croatian Science Foundation project-IP-2016-06-4602 to GŽ.

Ciljevi istraživanja: U modelu jedne umjerene traumatske ozljede mozga (engl. traumatic brain injury, TBI) odrediti pojavnost i opseg izražaja TDP-43 u ipsilateralnom i kontralateralnom korteksu i hipokampusu, usporediti TDP-43 subcelularni izražaj nakon jedne umjerene TBI i blage repetitivne TBI (engl. repetitive mild traumatic brain injury, rmTBI), te odrediti povezanost TDP-43 izražaja s biljezima sinaptogeneze i upale nakon TBI. Materijal i metode: Metodom lateralne ozljede tlakom tekućine (engl. lateral fluid percussion injury, LFPI) izazvana je jedna TBI umjerene jakosti u miša, dok je metodom pada utega izazvana rmTBI. Izražaj proteina od interesa analiziran je u citoplazmatskim i nuklearnim frakcijama ipsilateralnog i kontralateralnog korteksa i hipokampusa u životinja LFPI skupina, te frontalnom korteksu i hipokampusima rmTBI miševa. Histološkim tehnikama praćene su moždane promjene, a imunofluorescentnim tehnikama subcelularna lokalizacija TDP-43 te povezanost s biljezima upale i sinaptogeneze. Rezultati: Ipsilateralni korteks LFPI životinja najoštećenija je moždana struktura koju prati dezintegracija bijele tvari. U ipsilateralnom korteksu pronađena je translokacija TDP-43 u citoplazmu prvog i trećeg dana nakon LFPI u neuronima i mikrogliji. U ipsilateralnom hipokampusu uočen je pad izražaja nuklearnog TDP-43 trećeg dana od jedne umjerene TBI. Četrnaest dana nakon jedne umjerene LFPI, ali i rmTBI uočen je porast izražaja nuklearnog TDP- 43 u ipsilateralnom, odnosno frontalnom korteksu. Detektiran je porast izražaja fragmentiranog TDP-35 te značajan porast izražaja fosforiliranih oblika P-TDP-43 i P-TDP-35 u ipsilateralnom hipokampusu četrnaest dana nakon jedne LFPI, ali ne i u hipokampusu rmTBI životinja. U ipsilateralnom korteksu LFPI životinja uočena je povezanost biljega upale NF-κB i TDP-43 trećeg dana u području uz najveće oštećenje te povećan izražaj biljega iNOS prvog dana uz povezanost s TDP-43 u području najvećeg oštećenja. Pad izražaja PSD-95 uz gubitak neurona ipsilateralnog korteksa praćen je pozitivnom korelacijom s TDP-43 prvog, trećeg i četrnaestog dana od LFPI. Porast izražaja proteina APP uočen je subakutno samo u rmTBI skupini životinja, a proteina P-tau samo u LFPI miševa. Zaključak: Jedna umjerena LFPI uzrokuje deregulaciju TDP-43 u ipsilateralnom korteksu i hipokampusu koja utječe na pojedine biljega upale i sinaptogeneze. Navedene promjene ovise o zadobivenoj traumi mozga i od potencijalne su važnosti u farmakoterapiji TBI.

Introduction/Objectives: Traumatic brain injury (TBI) is the leading cause of death and disability that significantly burdens the health care system and society. Primary brain injury is irreversible, while secondary TBI includes pathophysiological processes that evolve over time and lead to memory and cognitive impairment. Along with the genetic predisposition, TBI is considered a risk factor for the development of Alzheimer's disease (AD). This study aimed to investigate changes in the expression of amyloid-beta precursor protein (APP) and Tau, two major proteins involved in the AD pathogenesis, in the brain cortex of C57BL/6 mice at 14 days following single moderate TBI. Participants, Materials/Methods: Lateral fluid percussion injury was used to induce a single moderate TBI over the left parietal cortex. Animals were sacrificed 14 days after injury and their cortices were prepared for Western blot or immunohistological analyses. Mice subjected to sham injury were used as a control group and were sacrificed 1 day after the sham procedure. Results: Loss of cortical NeuN positive neurons was evident in the area of the greatest brain damage but also in the pericontusional area. Subacute cortical changes in the APP expression were not detected, but cells positive to phosphorylated Tau protein were observed in the damaged cortex of brain-injured mice. Conclusions: Our preliminary results showed that single brain trauma causes loss of cortical neurons and Tau phosphorylation, without evident APP changes at 14 days post-injury. This work was supported by the University of Rijeka, Croatia, project number uniri-biomed-18-199 to K.P.