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Invasive alien crayfish threaten freshwater biodiversity and native crayfish fauna. However, the invasion success of an otherwise successful crayfish invader may be compromised by local environmental conditions or presence of local pathogens which can affect invader’s physiology, immune status, health, and fitness. Here we report the occurence of idiopathic necrotizing hepatopancreatitis and changes in the immune response in the signal caryfish, one of the most successful freshwater invaders, in a recently invaded Korana River in Croatia. We sampled and histologically analysed 73 signal crayfish individuals collected in three consecutive years (2018-2020) and used several standard immune parameters (encapsulation response, hemocyte count, phenoloxidaze activity and total prophenoloxidaze) to: i) compare immune response of the signal crayfish along its invasion range, ii) analyze effects of specific predictors (water temperature, crayfish abundance and body condition) on its immune response changes. Our results show very high prevalence of lesions in hepatopancreas of signal crayfish (>91%), with 14% of individuals displaying heavy and 51% moderate necroses and/or inflammation in the tubular and interstitial structures of the organ. Heavy histopathological changes in hepatopancreas were correlated with lower organosomatic condition. Immune response exhibited significant differences along the invasion range of the signal crayfish and was mostly affected by water temperature and population abundance. Obtained results offer a baseline for elucidating the role of immunocompetence in the signal crayfish invasion in the Korana River, while aetiology of necrotizing hepatopancreatitis needs to be further investigated to determine its potential impact on signal crayfish invasion success.

Subfosilno drvo ili abonos je drvo koje je više stotina ili tisuća godina odležalo na dnu močvare, rijeke, jezera ili mora pod naslagama šljunka, blata i mulja u anoksičnim uvjetima. Karakteristična crna boja abonosa potječe od kemijske reakcije tanina iz drva i željeza iz vode. Subfosilno drvo ima promijenjena fizikalna i mehanička svojstva, zbog specifičnih uvjeta u kojima je nastajalo. Recentno i subfosilno drvo razlikuju se u sastavu, jer se u subfosilnom drvu tijekom godina smanjivao sadržaj ekstraktivnih tvari i fenolnih spojeva, te se također smanjivao udio ugljikohidrata. Uspoređivanjem sadržaja anorganskih elemenata u subfosilnom i recentnom drvu iste vrste dokazalo se da subfosilno drvo ima veći ukupni sadržaj željeza, kalcija i magnezija, dok recentno drvo ima veći ukupni sadržaj kalija. Sadržaj celuloze u subfosilnom drvu manji je nego u recentnom, dok je sadržaj lignina puno veći u subfosilnom drvu. Gustoće recentne i subfosilne hrastovine slične su, ali pokazalo se da povećanjem starosti abonosa dolazi do povećanja njegove gustoće. Subfosilno drvo ima širok spektar upotrebe, može se koristit pri izradi jedinstvenih skupocjenih ukrasa, namještaja ili glazbenih instrumenata. Subfossil wood or abonos is wood that has lain for hundreds or thousands of years at the bottom of a swamp, river, lake or sea under deposits of gravel, mud and silt in anoxic conditions. The characteristic black color of abonos comes from the chemical reaction of tannins from the wood and iron from the water. Subfossil wood has changed physical and mechanical properties, due to the specific conditions in which it was formed. Recent and subfossil wood differ in composition, because the content of extractive substances and phenolic compounds in subfossil wood has decreased over the years, and the proportion of carbohydrates has also decreased. By comparing the content of inorganic elements in subfossil and recent wood of the same species, it was proven that subfossil wood has a higher total content of iron, calcium and magnesium, while recent wood has a higher total content of potassium. The cellulose content in subfossil wood is lower than in recent wood, while the lignin content is much higher in subfossil wood. The densities of recent and subfossil oak are similar, but it has been shown that increasing the age of abonos leads to an increase in its density. Subfossil wood has a wide range of uses, it can be used to make unique expensive decorations, furniture or musical instruments.

Human, mammalian, brain development is a very complex process and starts shortly after gastrulation. Human in vitro models for brain development as well as pathological changes were published in last few years, eliminating the ethical concerns that accompany the study of aborted foetal material. One of the recent approaches is the development of induced pluripotent stem cell (iPSC) technology and different types of organoids, which can be applied to any species. In our previous work we developed iPSCs from an adult individual with constitutional mosaicism for Down Syndrome (DS) (1). From those iPSCs, cerebral organoids were grown and corticogenesis and Alzheimer-like pathology in vitro were studied (2). Mixed cerebral (2), as well as midbrain (unpublished) organoids were grown in vitro from the isogenic, trisomic and disomic iPSCs. Organoids were analysed histologically by IMARIS software. Foetal human brains were stained with the same combinations of antibodies and the data were compared. Anonymized post-mortem brain samples were obtained from the Brain Bank of the Croatian Institute for Brain Research (CIBR). Isogenic cerebral organoids expressed the markers of all six cortical layers (Reelin, FOXG1, SATB2, TBR1, Ctip2 and Brn2) after 100 days differentiation in vitro (DIV). At the early time point of DIV30 the majority of cells were Nestin, SOX2, PAX6 positive. At that time point neurogenesis had already started, so we observed TRBR2, DCX, MAP2 and TUBB3 positive cells in disomic and trisomic organoids. Following further neuronal differentiation of cerebral organoids we observed decreased number of neural stem cells and increased number of neuronal markers specific for all six layers. On the other hand, midbrain organoids expressed markers specific for midbrain. At the early time points midbrain organoids expressed neuronal stem cells markers, similar to cerebral organoids, but with organoid maturation we found midbrain specific cells such as NURR1 and OTX2 specific for the early stages, and later Dopamine β Hydroxylase, NET and TH. Finally, all those markers were compared with expression in foetal human brain from individuals with Down Syndrome as well as euploid controls age, sex and brain region matched. Our data showed that isogenic cerebral organoids grown from iPSCs are good tool for multidisciplinary studies. In our group we study corticogenesis, synaptogenesis, pathology and possible drug treatments.

Background: Specific cell types from people with Down syndrome (DS) show faster accumulation of DNA damage and epigenetic senescence marks. Causative mechanisms remain un-proven ranging from amplified chromosomal instability to actions of chromosome 21 genes. Plasma immunoglobulin G (IgG) glycosylation profiles are established as a reliable predictor of biological and chronological ageing. Methods: We performed IgG glycan profiling of n=246 individuals with DS (208 adults and 38 children) clinically characterised for co- morbidities, from three European countries, and compared these to age-, sex- and demography- matched general populations. Results: We uncovered very significantly increased IgG glycosylation ageing marks associated with DS, beginning in early childhood. Average levels of IgG glycans without galactose (G0) and those with two galactoses (G2) as a function of age in persons with DS corresponded to levels detected in 19 years older euploid individuals. Sub-cohorts of DS without any known co-morbidities from different countries showed a similar result, eliminating DS- linked autoimmune diseases, frequent infections, or Alzheimer’s dementia as indirect causes of accelerated biological ageing. Importantly, no difference in slope of the curve relating IgG- glycan-age to chronological age was observed for any of the 3 analysed populations, discouraging the amplified-chromosomal-instability explanation. Remarkably, IgG-glycan profiles of a child with DS caused by a short segmental duplication of only 31 genes on chromosome-21 (DS-critical-region) had values like age-matched whole-chromosome-trisomy- 21 children, pointing to a causative contribution of one or more of the 31 genes. Conclusion: This shifts the paradigm from an (essentially untreateable) “amplified instability” mechanism, to a specific gene-overdose-driven cause of a progeria-like syndrome, opening possibilities for therapeutic amelioration strategies. We generated an induced pluripotent stem cell (iPSC) model of this child with partial trisomy 21, and editing experiments using CRISPR-Cas9 are underway in order to dissect the causative overdosed genes and mechanisms responsible for the accelerated ageing in DS.

Objectives: To describe neuronal differentiation from the disomic (D21) and trisomic (T21) isogenic human induced pluripotent stem cells (iPSCs) and neural stem cells (NSCs) derived from persons with partial and full trisomy 21, and mosaic Down syndrome (DS). Methods: NSC were generated form the isogenic iPSCs following published protocol (1) and expanded in our laboratory. NSCs, between passage 5 and 10 were seeded on the commercially available mouse astrocytes and differentiated to the mature neurons 100 days in vitro (DIV) (2). Moreover, cerebral organoids were generated following published protocol (3) with our modification (4) and fixed at 30, 50, 70 and 100 DIV and analysed by immunohistochemistry. Results: In our previous work we published neuronal differentiation from EBs D3 and 47-1 (5). Here we find that Tubb3 positive cells show a specific neuronal phenotype. Disomic cells showed normal neuronal morphology, on the other hand trisomic cells showed a very specific cellular dysmorphology: neurons were smaller, shorter with thicker processes, and abnormal branching patterns. The same pattern we have observed in neurons derived form iPSCs in monolayer as well as in cerebral organoids. Both, neurons in monolayer, as well as in cerebral organoids expressed all neuronal markers: pan neuronal markers (Dcx, MAP2. Tubb3, SMI, 3R-Tau) and cortical layer specific markers (Reelen, Brn2, Ctip2, SATB2, TBR1 and TBR2). Conclusion: Trisomic cells could differentiate to mature, synaptic active neurons but morphology of trisomic cells shows a unique and specific dysmorphology pattern.

Granični poremećaj ličnosti ozbiljan je mentalni poremećaj za koji su karakteristične emocionalne, bihevioralne i interpersonalne smetnje koje rezultiraju velikim poteškoćama u profesionalnom, akademskom i socijalnom funkcioniranju te povećanim rizikom od smrtnosti. Kako je za ovaj poremećaj ličnosti karakteristična jedinstvena interpersonalna osjetljivost, odnosno povećana vulnerabilnost na međuljudske stresore, glavni simptomi često su izričito interpersonalne prirode. Te poteškoće u interpersonalnom segmentu povezane su s problemima u romantičnim odnosima. U objašnjavanju disfunkcije romantičnih odnosa osoba s graničnim poremećajem ličnosti najznačajnije su teorija privrženosti i Linehanova biopsihosocijalna teorija, kao i biološki pristupi koji naglašavaju abnormalnosti neurotransmiterskih sustava te kognitivni pristup u čijem su središtu zanimanja kognitivne sheme pri objašnjavanju disfunkcionalnih kognicija osoba s graničnim poremećajem. Istraživanja koja se bave proučavanjem graničnog poremećaja ličnosti u kontekstu romantičnih odnosa usmjerena su na karakteristike osoba s tim poremećajem, kao i karakteristike njihovih romantičnih odnosa. Istraživanja karakteristika samih romantičnih odnosa usmjerena su na duljinu odnosa i broj romantičnih partnera. S druge strane, analize karakteristika osoba s graničnim poremećajem ličnosti koje se odražavaju na romantične odnose usredotočene su na 1) emocionalno zadovoljstvo; 2) afektivnu nestabilnost (osjetljivost na odbacivanje/strah od napuštanja, nestabilno raspoloženje, hostilnost); 3) stil privrženosti i 4) stil komunikacije. Rezultati istraživanja pokazuju kako su za osobe s graničnim poremećajem ličnosti, u usporedbi s osobama bez tog poremećaja, karakteristične mnogobrojnije i kratkotrajnije romantične veze, manje emocionalno zadovoljstvo, veća razina hostilnosti te veća vjerojatnost nesigurne privrženosti i pasivno-agresivnog stila komunikacije.

Signalni rak predstavlja opasnost za autohtone vrste rakova zbog velike agresivnosti i dominacije u kompeticiji za resurse. Kako je hepatopankreas najvažniji energetski organ signalnog raka, promjene u njegovoj strukturi narušavaju i njegovu funkciju, a time i kondicijsko te cjelokupno zdravstveno stanje jedinke što može negativno utjecati na rast, reprodukciju i širenje ove vrste. U ovom radu prikupljene su i morfometrijski obrađene 73 jedinke signalnog raka, duž invazivnog areala u rijeci Korani. Jedinkama su izolirani hepatopankreasi te izrađeni trajni histološki preparati koji su analizirani svjetlosnim mikroskopom. Dobiveni rezultati pokazuju da 64% analiziranih rakova ima više od 25% tkiva hepatopankreasa zahvaćeno patološkim promjenama, pri čemu 14% životinja pokazuje teške promjene na više od 75% tkiva. Fokalna nekroza uočena je kod 53% jedinki, melanizacija kod 51%, a gram-pozitivne bakterije kod 44% jedinki. Ustanovljene histopatološke promjene indikatori su idiopatskog nekrotizirajućeg hepatopankreatitisa, čiji je uzročnik zasad još nepoznat. Sva histopatološka oštećenja zabilježena su s većom učestalošću u središtu invazivnog areala, gdje je gustoća jedinki veća, u odnosu na fronte širenja. Životinje s težim histopatološkim oštećenjima imale su značajno smanjeni hepatosomatski indeks, što ukazuje da ova bolest utječe na kondicijski status životinja, a time može utjecati i na daljnju brzinu širenja ove invazivne strane vrste. Signal crayfish threatens native crayfish populations due to its high aggression and dominance in competitive interactions. Since hepatopancreas is the main energy supply organ in crayfish, changes in its structure alter its function and affect its condition and overall health. This can consequently lead to negative impacts on animal growth, reproduction and range expansion. In this research, 73 individuals along signal crayfish invasion range in the Korana river were collected. Hepatopancreases were isolated, fixed histological slides were prepared and analyzed with the light microscope. Results of this experiment show that 64% of analyzed crayfish had more than 25% of tissue with pathological changes, with 14% of crayfish having more than 75% of tissue with heavy changes. Focal necrosis was observed in 53% of cases, while melanisation in 51% and Gram-positive bacteria in 44%. The aetiology of observed histopathological changes indicates idiopathic necrotizing hepatopancreatitis whose cause still remains unknown. All histopathological changes were more frequent in the invasion core, where population density is higher, opposed to invasion fronts. Heavy histopathological changes in hepatopancreas correlated with low hepatosomatic index which indicates that this disease affects the condition of the animal and may affect range expansion of this invasive alien species.

Introduction: Music is a very complex stimulus, with many different psychological and physiological effects. Although the ways it works on us remain largely elusive, music has been recommended as therapeutic tool for a variety of physical, psychological, cognitive, as well as social problems. In order to get a better understanding of music as medicine, we have reviewed a number of articles and collected information about music medical usage in the treatment of various disorders and conditions. Materials and Methods: PubMed base has been searched for English-language articles of interest. Selected topics associated with music included stress, anxiety and depression, sleep disorders, as well as neurorehabilitation with focus on specific medical conditions. Results: Various studies demonstrated that listening to music can improve sleep, by promoting relaxation or serving as a distraction, and it has been recommended for treatment of both acute and chronic sleep disorders. Music can be used to alleviate anxiety, depression and pain, or for motivation and energizing. Positive influence of music on concentration, mood, stress reduction, as well as heart rate and blood pressure, have been also reported. In addition, various subtypes of music therapy can be a useful tool in neurorehabilitation of disorders such as stroke, Alzheimer’s and Parkinson’s disease, due to beneficial effects on patient’s motor, cognitive and psychological disturbances. Conclusion: Unlike drugs or medical procedures, music needs no government approval or clinical trials— it is inexpensive and simple way to improve the quality of people's lives and cannot hurt even if it fails to help.

Članak upozorava na barjak kao važan svadbeni rekvizit i simbol na području čitave Like. Posebna pozornost posvećena je lokalnim razlikama u ukrašavanju barjaka. Autorica upućuje i na promjenu funkcije barjaka, odnosno stapanje (ili pretapanje) njegova značenja kao simbola svadbe i kao simbola etničkog identiteta.