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The objective of this research was to isolate and identify lactic acid bacteria living in the digestive tract of the queen scallop Aequipecten opercularis and further test it as a probiotic culture on the growth of the bivalve during one month in captivity. Classic microbiological techniques (API and MALDI TOF) were used and four different strains of Lactiplantibacillus and one Lactococcus were identified and further analyzed, namely: Lactiplantibacillus plantarum I, Lactiplantibacillus plantarum 1, Lactiplantibacillus plantarum 2, Lactococcus lactis, and Lactiplantibacillus brevis. The isolated cultures were further tested for the highest antimicrobial activity towards the most common marine pathogens and survival at different temperatures and pH levels. The strain Lactiplantibacillus plantarum I had the best results, with the highest antimicrobial activity (77–85% pathogen growth inhibition) and the best safety standards showing no antibiotic resistance, and no ability to synthesize biogenic amine and degrade red blood cells. Considering all the abovementioned characteristics, the strain Lpb. plantarum I was further tested on the growth and health status of the bivalve Aequipecten opercularis during a month of cultivation in captivity. The results showed that the incorporation of Lpb. plantarum I led to a decrease in the quantity of test microorganisms in the bivalve and an increase in both the growth rate weight and growth rate length of the queen scallop A. opercularis. Furthermore, the scallops fed with the algae culture + Lpb. plantarum I gained significantly higher meat yield (33.15 ± 2.63%) compared with the control scallops fed only with the algae culture (29.66 ± 2.87%). To conclude, the results from this research indicate that including Lpb. plantarum I as a dietary supplement could enhance growth performance and serve as a feasible approach to reduce pathogen levels while cultivating A. opercularis in captivity.

Seasonal changes in environmental parameters affect the metabolic activities and physiology of bivalves. The aim of this study was to determine the impact of environmental parameters on the queen scallop Aequipecten opercularis (Linnaeus, 1758). Scallops were sampled in October 2020 at the Kamenjak Peninsula area, Northern Adriatic. Their culture was maintained for a period of one year in tanks with flowing seawater exchange. To assess the seasonal impact of temperature, pH, and oxygen levels on growth and physiological status, queen scallops were sampled from the tanks four times a year and morphometric parameters, somatic indices and condition index were measured. Seasones had the greatest impact on the scallop's soft body weight, followed by the bivalve total weight, and finally on the morphometric characteristics. The highest condition index was recorded in autumn (94.04%) and the lowest in summer (46.67%). Somatic indices varied during the seasons ; the muscle index was lowest in summer (3.81%) and the highest in winter (5.96%) while the gonadosomatic index is lowest in autumn (18.01%) and highest in spring (34.06%). The wet soft body weight and the condition index are dependent on the seawater temperature and pH, while morphometric parameters were less influenced by environmental factors.

The DNMT3B gene codes for DNA methyltransferase 3b (DNMT3b), a protein required for genome-wide de novo methylation during the early stage of embryonic development. Evidence suggested that impaired DNA methylation could be a risk factor for congenital heart defects (CHD). Down syndrome (DS) is one of the most common chromosomal abnormalities associated with congenital heart defects (CHD), with approximately 40 – 60% of cases showing cardiac defects. The most common CHD phenotypes in DS are congenital malformations of cardiac septa, such as atrioventricular septal defects (AVSDs), ventricular septal defects (VSDs), atrial septal defects (ASDs), and tetralogy of Fallot (TOF). The purpose of this study was to analyze the frequency of the phenotype of CHD as well as allele and genotypes association of functional DNMT3B promoter polymorphism −579 G > T (rs1569686) on the development of CHD in DS. The case-control study included 102 CHD+ Down syndrome cases and 88 non- syndromic CHD cases. The median of the ages for CHD+DS cases was 2 years [0 - 27] and for the control group was 7 years [0 - 32]. Demographic data and phenotype of CHD were collected from medical records of participants after parents and guardians gave their written consent. Genomic DNA was isolated from different types of tissue using a commercial kit and quantified by spectrophotometry. Genotype analysis was performed by PCR-RFLP method. Statistically higher frequencies of AVSD and VSD were detected in a group of CHD+DS than controls (P=0.0044 ; P=0.0225). Study results showed a statistically lower frequency of the DNMT3B rs 1569686 TT genotype (OR = 0.40 ; 95% CI: 0.18 – 0.86 ; P=0.020) in CHD+DS cases compared with controls. AVSD and VSD were the most common CHD phenotypes in DS, which were consistent with the literature. Lower frequencies of DNMT3B (rs1569686) TT genotype in- group CHD+DS suggested that those genotype could decreased risk of CHD in DS individuals. Further ongoing studies will better explain those results.

Uvod: Europski kvalifikacijski okvir predstavlja poveznicu nacionalnih kvalifikacijskih sustava zemalja članica Europske unije s ciljem usporedbe razine kvalifikacija među članicama kako bi se ujednačilo tržište rada unutar EU ali i poticalo građane na cjeloživotno obrazovanje bilo u formalnom, neformalnom ili informalnom učenju. Razrada: Prema navedenom, nacionalni kvalifikacijski sustavi trebaju biti povezani s razinama Europskog kvalifikacijskog okvira (EQF) prema transparentnoj metodologiji uz sudjelovanje svih zainteresiranih dionika, što se može osigurati uz pristup informacijama i smjernicama o Europskom kvalifikacijskom okviru i nacionalnim kvalifikacijskim sustavima. Republika Hrvatska je stoga donijela Zakon o Hrvatskom kvalifikacijskom okviru (HKO) koji predstavlja uređeni sustav kvalifikacija kojim se osigurava pristupanje, stjecanje, prohodnost i kvalitetu kvalifikacija osiguravajući time povezivost s EQF te time posredno i s ostalim nacionalnim kvalifikacijskim sustavima. Kako bi se osigurala povezivost ustrojava se Registar HKO koji se sastoji od tri podregistra: skupova ishoda učenja (stečene kompetencije), standarda zanimanja (popis poslova i kompetencija u određenom zanimanju) i standarda kvalifikacija (sadržaj i struktura određene kvalifikacije zasnovane na objedinjenim skupovima ishoda učenja). Zaključak: Obzirom na ciljeve projekta „Razvoj i unapređenje studijskih programa sukladno HKO-u na Sveučilištu Jurja Dobrile u Puli“ rad analizira dokumente vezane uz standard zanimanja medicinske sestre/tehničara opće njege (razina 4.2 HKO), standard zanimanja prvostupnika/ce sestrinstva (razina 6 HKO) i magistra/ce sestrinstva (razina 7.1 HKO) gdje rezultati upućuju na postojanje određenog stupnja kompleksnosti između razina standarda zanimanja obzirom istima se definiraju ključni poslovi i kompetencije, te posljedično znanja, vještine i razine samostalnosti u obavljanju zadaća te ujedno i osigurala usporedivost zanimanja u sestrinstvu s ostalim zemljama članicama Europske unije.

The Mediterranean Sea displays high species diversity and represents a key area for study of the impacts of natural and anthropogenic changes on biodiversity and its consequences on ecosystem. Habitat destruction, over-fishing, contaminants, eutrophication, introduction of alien species, and climate changes are causing increasingly evident changes in community structure and biodiversity of this warm and miniaturized ocean. Yet, Mediterranean fish biodiversity is undergoing rapid changes due to the rise of water temperature and the increasing success of thermophile biota. In this context, the analysis of continuous long-term and multiparametric data offers a new and promising opportunity to ecologists to understand those processes which regulate marine biodiversity and may also contribute to define possible strategies of conservation and ecosystem-based management. Coastal-cabled video observatories such as the Observatory of the Sea (OBSEA ; www.obsea.es) off Vilanova i la Gertrù (Spain) have been indicated as innovative andchallenging technological tools for monitoring fish assemblages in marine systems. In addition, these devices can be successfully employed to monitor and help to understand the responses of fish species to both, environmental drivers and human stressors, as also invoked by the EU Marine Strategy Framework Directive. These multiparametric intelligent platforms represent one of the key elements for Blue Growth development policies. Cabled, fixed and permanent observatories are being worldwide-distributed within growing geographic networks. They provide long-term monitoring of several characteristics (oceanographic, biological etc.) of the seabed and water column at various depths, in a synchronous and highly integrated fashion. Here, we describe how are the video monitoring actions introduced and applied at the OBSEA within the framework of RESBIO Spanish National Project and the resulting experience transferred to a cabled video station in Šibenik, Croatia.

Procjena učestalosti pojavljivanja gospodarski važnih vrsta riba u estuariju rijeke Krke, uvala Martinska nasuprot Šibenika provedena je od siječnja do travnja 2018. godine podvodnom video kamerom s kabelskim napajanjem. Na osnovu analize videozapisa utvrđeni su: postotak zastupljenosti gospodarski značajnih vrsta u odnosu na ostale prisutne vrste riba, ukupan broj gospodarski važnih vrsta riba i učestalost pojavljivanja njihovih jedinki po mjesecima. Utvrđeno je ukupno 36 vrsta, od čega su 23 gospodarski značajne (63, 9 %). Analize bogatstva ihtiofaune pokazale su varijacije tijekom četiri mjeseca. U cilju dobivanja detaljnih podataka o učestalosti pojavljivanja svake pojedinačne vrste i stanju njihovih populacija, bilo bi potrebno provoditi ovakav nelovni monitoring kroz duži vremenski period.

Nukleotidni slijed mitohondrijskog gena za podjedinicu 1 citokrom oksidaze (COI ili COXI) jedan je od najčešće korištenih markera za identifikaciju vrsta i studije populacijske genetike. Cilj ovog preliminarnog istraživanja bilo je utvrđivanje razlika u distribuciji COI haplotipova u dagnje Mytilus galloprovincialis duž istočnog dijela Jadrana. Na različitim lokacijama očekivana je različita raspodjela haplotipova. Po pet dagnji uzorkovano je na šest različitih lokacija. Ukupna genomska DNA izolirana je iz tkiva škrga. Fragmenti COI DNA umnoženi su lančanom reakcijom polimerazom (PCR). Amplificiranim uzorcima mtDNA određeni su nukleotidni slijedovi. Dobiveni su slijedovi COI haplotipova dužine 602 nukleotida. Analizom je ustanovljeno 8 različitih haplotipova, različitih učestalosti i rasprostranjenosti. Najčešći haplotipovi HapA3 (33, 33% svih sekvenci) i HapA4 (26, 67%) prisutni su na svim postajama. Drugi, manje česti haplotipovi detektirani su kako slijedi: HapA1 i A5, s 10%-tnom učestalošću prisutni su na po 2 postaje, HapA2 (10%) na 3 postaje i HapA6, A7 i A8 prisutni su s po jednom kopijom (3.33%) samo na po jednoj postaji. Ova preliminarna studija, iako vrlo ograničena, otkrila je veliku varijabilnost i nejednaku raspodjelu haplotipova duž istočnog dijela Jadrana. Distribucija haplotipova na različitim postajama mogla bi biti povezana s različitim fizikalno-kemijskim svojstvima okoliša, ali priroda takvih veza može se odrediti tek nekom većom i detaljnijom studijom.

Legally prescribed ecological data monitoring has not been established at most of marine Natura 2000 areas in Croatia, and therefore legislation of ecosystems management and protection is not completely fulfilled. For the first time, to the best of our knowledge, a coastal time-lapse imaging survey has been carried out targeting the Šibenik coastal fish community (off Krka River, 5 m depth). Temperature and salinity data were collected concomitantly, both parameters being relevant to fish presence and abundance. One image per hour was collected during daytime continuously from January to March 2018, such collection is still being continued today. A total of 7 311 time-lapse images were analyzed for species classification and 288 Fish Forum Book of abstracts animal counting (epibenthic invertebrates included). Forty-one taxa were detected with fish prevalence, 70.73 percent of all recorded species. The rest being echinoderms (12.25 percent), cephalopods (7.3 percent), crustaceans (2.43 percent), anthozoans (2.43 percent), reptilians (2.43 percent) and birds (2.43 percent). Relevant observations were made of flagship species such as bluefin tuna, Thunnus thynnus (Linnaeus, 1758) and loggerhead sea turtle, Caretta caretta (Linnaeus, 1758). The latter was observed in both hibernation and swimming modes. Species richness, calculation of biodiversity, and degree of coverage of benthic invertebrates were also obtained as proof of concept of the validity of such coastal monitoring actions for large and growing EU infrastructural assets such as the Expandable Seafloor Observatory (OBSEA) and the European Multidisciplinary Seafloor and water column Observatory (EMSO) testing sites. Our data support the need for permanent video-based technical development of the European Strategy Forum on Research Infrastructures (ESFRI) to increment its social projection.

Posljednjih godina su u Jadranu utvrđene nove vrste riba u različitim razvojnim stadijima. Radi se pretežno o termofilnim vrstama, koje preferiraju toplija mora. Nadalje, vrste uobičajene za južni dio Jadrana pomiču se sjevernije te su sve prisutnije u ribarskim lovinama tih područja. Stručnjaci objašnjavaju kako se zbog povećane brojnosti novih vrsta, Jadran nalazi pod utjecajem procesa „tropikalizacije“. Porast temperature i promjene u kemijskom sastavu mora pomažu uspostavljanju njihovih populacija. Određene vrste utvrđene su jednom ili nekoliko puta te još uvijek nije izvjesno jesu li stalni stanovnici ili samo povremeni gosti Jadrana. Putevi unosa brojnih vrsta još su uvijek nepoznati. Mnogi od njih su „lesepsijski migranti“, vrste koje su migrirale iz Crvenog mora kroz Sueski kanal. Osim ovim putem, neke su vrste unesene balastnim vodama, pomorskim transportom, bijegom ili uvođenjem novih vrsta u marikulturu. Kao putevi širenja spominju se i morske struje, koje s Otranta, uz nakupine smeća, najvjerojatnije donose i različite razvojne stadije novih vrsta. Za determinaciju novopridošlih vrsta te utvrđivanje njihovoga zemljopisnog podrijetla, uz klasične metode se sve češće rabe i molekularne. U ovom su radu prikazane nove vrste riba u Jadranu, promjena prostorne distribucije već prisutnih, mogući učinci na morski ekosustav te potencijalna mogućnost gospodarskoga iskorištavanja pojedinih vrsta.

Pojam održivoga razvoja u posljednje se vrijeme uvodi u sva područja ljudskoga života. Njime se bave svjetske organizacije, političari, ekonomisti, udruge civilnoga društva, obrazovne institucije i svi koji smatraju da mogu utjecati na održivi razvoj. Koncept održivoga razvoja počiva na ravnoteži nekoliko sustava u društvu: gospodarskog (ekonomskog), ekološkog (prirodnog) i društvenog (socijalnog). Čovjek svojim (ne)pažljivim ponašanjem prema prirodi utječe na okoliš. Pitanje očuvanja okoliša poprima široke razmjere jer utječe na život i zdravlje ljudi. Knjiga Odgoj i obrazovanje za razvoj danas i sutra. Ekološke i društvene paradigme koncepcijski je i sadržajno podijeljena u dva dijela: ekološki i društveni. Ekološkom paradigmom održivoga razvoja u knjizi daje se pregled stanja utjecaja na okoliš s obzirom na ljudsku populaciju, energiju i količinu emisije ugljičnoga dioksida. Preporuke za smanjenje utjecaja na okoliš temelje se na suvremenim stručno-znanstvenim smjernicama koje, između ostaloga, predstavljaju inicijativu za promicanje odgoja i obrazovanja za (održivi) razvoj. Društvena paradigma odgoja i obrazovanja za razvoj (engl. education for development) posebno je područje odgoja i obrazovanja i srodni pristup odgoju i obrazovanju za ljudska prava. U knjizi se ova paradigma razmatra u kontekstu odgoja i obrazovanja za održivi razvoj (engl. education for sustainable development) s posebnim osvrtom na odgoj i obrazovanje za ljudska prava i demokratsko građanstvo, odnosno, građanski odgoj i obrazovanje. Koncept odgoja i obrazovanja za razvoj u središte interesa dolazi početkom 70-ih godina dvadesetoga stoljeća u sklopu proglašenja Desetljeća razvoja Ujedinjenih naroda, posebice nakon što se uvidjelo da gospodarska i tehnička pomoć bez odgoja i obrazovanja nije dovoljna za poticanje napretka nerazvijenih zemalja. Pokreću ga međunarodne institucije i entuzijasti u odgoju i obrazovanju. Problemi odgoja i obrazovanja za održivi razvoj po svojoj su prirodi interdisciplinarni. U knjizi se razmatraju analize, stavovi i istraživanja autora koji većinom zaključuju da odgoj i obrazovanje za održivi razvoj jača i razvija sposobnost pojedinaca, skupina, zajednica, organizacija i zemalja da djeluju u korist održivoga razvoja. Takav pristup obrazovanju može učiniti pomak u načinu razmišljanja i djelovanja ljudi kako bi svijet učinili sigurnijim, zdravijim i prosperitetnijim, poboljšavajući time kakvoću života na zemlji. Odgoj i obrazovanje za održivi razvoj omogućuje stjecanje znanja, vještina, stavova i vrijednosti potrebnih za oblikovanje održive budućnosti te uključivanje ključnih pojmova etike održivosti u odgojno-obrazovni proces. On ulazi u sve društvene kontekste (obitelj, predškolske ustanove, školu, radno mjesto, lokalnu zajednicu), oblikuje odgovorne građane i promiče demokraciju dopuštajući pojedincu i zajednici uživanje u pravima i poštivanje odgovornosti prema održivom razvoju. Pravi problem s kojim se čovječanstvo danas suočava u smislu postizanja održivoga razvoja jest kako obrazovati i motivirati ljude da promijene temeljna ponašanja i aktivnosti koje su problematične i neodržive. U knjizi je dat pregled analize sadržaja europskih i hrvatskih normativnih i razvojno-strategijskih dokumenata u području odgoja i obrazovanja kojima se eksplicitno ili implicitno uređuje područje održivoga razvoja i odgoja i obrazovanja za (održivi) razvoj u kontekstu zaštite i promicanja ljudskih prava. Ističe se da središnje pitanje demokratskoga društva nije puko priznanje ljudskih prava, nego učinkovitost sustava njihove zaštite i promicanja. Knjiga daje pregled normativne osnove održivoga razvoja u Republici Hrvatskoj. U njoj su obrađeni strateški dokumenti iz područja održivoga razvoja i zaštite okoliša. Nacionalna normativna osnova uvođenja odgoja i obrazovanja za ljudska prava i njemu srodnih područja u odgojno-obrazovni sustav predstavlja ideju vodilju u promicanju odgoja i obrazovanja za (održivi) razvoj. Njegovo uključivanje može se pratiti uvođenjem odgoja i obrazovanja za ljudska prava u hrvatski odgojno-obrazovni sustav: od Nacionalnoga programa odgoja i obrazovanja za ljudska prava i demokratsko građanstvo (1999) do danas kada se Program međupredmetnih i interdisciplinarnih sadržaja građanskoga odgoja i obrazovanja za osnovne i srednje škole (2014) izvodi kao obavezan u osnovnim i srednjim školama Republike Hrvatske. Strukturne dimenzije stjecanja građanske kompetencije (ljudskopravna, politička, društvena, međukulturna, gospodarska i ekološka) u recentnijim hrvatskim obrazovnim dokumentima izdvojene su polazeći od prava i odgovornosti kojih je učenik nositelj kao član različitih zajednica: razredne i školske, lokalne i nacionalne, europske i međunarodne. Kurikulum odgoja i obrazovanja za održivi razvoj polazi od spoznaje o povezanosti globalnih promjena na Zemlji i njihova utjecaja na čovjeka i okoliš. Pitanja vezana uz održivi razvoj, posebice proces odgoja i obrazovanja za održivi razvoj, često su kontroverzna i složena stoga je važno znati kako riješiti kompleksne situacije. U knjizi se navode mogućnosti nastavnoga rada u odgoju i obrazovanju za održivi razvoj kao što su: rasprava, simulacije problema, eksperimentiranja, terenska nastava, rad na projektima i uključivanje u akcije na temu održivoga razvoja. U njoj je istaknuta važnost novih pristupa podučavanju, pozicioniranje učenika u središte obrazovnoga procesa, kao i uloga profesionalnih kompetencija učitelja pri planiranju, programiranju, provedbi i vrednovanju postignuća u obrazovanju za održivi razvoj, jer upravo o njihovim kompetencijama ovisi uspješnost implementacije održivoga razvoja u praksu. Pristup u podučavanju treba osigurati kontekst u kojemu učenici mogu razvijati svoje ideje, vrijednosti te razumijevanje i promišljanje životne stvarnosti. Učenicima treba omogućiti aktivno sudjelovanje u izgradnji vlastitoga znanja. Na kraju knjige čitatelju se nude odabrani primjeri dobre prakse (za prvi i šesti razred osnovne škole, te za treći razred srednje škole) kao poticaji za izvođenje nastavne djelatnosti. Budući da su u svijetu, a osobito u nas, usporedbe ekološke i društvene paradigme u području odgoja i obrazovanja za (održivi) razvoj malobrojne, teorijska razmatranja i rezultati ovakvih komparativnih analiza pridonose prirodnim i društvenim znanostima u kontekstu interdisciplinarnosti.

Leucocyte depletion improves the quality of stored blood units. We have studied its role on blood viscosity. Viscosity of CPDA-1 blood units was measured in a Couette viscometer at shear rates of 94.5 and 0.1 s-1 prior to and following filtration with the Leukotrap® A1 system on day 0 and after 21 days at +4°C. On day 0, high but not low shear viscosity was significantly decreased. The red blood cell morphology was unaffected. On day 21, blood viscosity was increased similarly for unfiltered and filtered samples at both shear rates. The echinocytosis observed after storage correlated with the increase in viscosity. Leucocyte depletion is associated with a decrease in high shear viscosity. This effect is, however, completely lost after 21 days.

P53 family genes play a key role in cell cycle control and maintenance of genetic stability. The family consists of three genes: p53, p63 and p73 ; all sharing a remarkable structural and functional similarity. The presence of genotoxic pollutants in the sea causes changes at molecular and cellular levels in various organisms. Such stressful conditions change the expression levels and activity of various genes including the p53 family. One of the well recognized bioindicator organisms of genotoxic stress is a common blue mussel (Mytilus sp.) because it is a sessile, filter feeding marine animal with a worldwide coastal water distribution. Standard molecular biology methods, including genomic DNA extraction, polimerase chain reaction, cloning and sequencing, were used to obtain the genomic sequence of p53 family homolog in Mytilus galloprovincialis. The exon/intron architecture and deduced protein sequences were analysed and compared to p53 family sequences from other organisms found in the GenBank database and revealed a conserved gene structure among different organisms. Our analysis reinforced the previous conclusion that invertebrates have only one gene of the p53 family which radiated into three genes only in vertebrates. The presented genomic structure of Mytilus galloprovincialis p53 family homolog shows to be most similar to p63 in vertebrates and supports the notion that p63 is the ancestral gene of the p53 family. This is in accordance with our previous work performed with partial cDNA and genomic sequences of Mytilus galloprovincialis.

Seawater salinity changes and thermal stress have been considered to be among the most important determinants (and possible stressors) of organism distribution in the rocky intertidal zone. Due to the changes in physical characteristics of water, the marine environment can be stressful to its inhabitants. Some of them, including the mussels, can be used as tracers of environmental conditions and pollution. The aim of present study was to determine the freshwater influx and evaluate the mussel Mytilus galloprovincialis shells as indicators of natural environmental conditions by comparing δ13C of shell carbonate layers with seawater salinity and temperature values measured over the period 1998-2007 on different locations included in the Croatian National Biomonitoring Programme “Project Adriatic”. Our results show that the salinity was relatively constant in the open sea, with typical values in the northern and southern Adriatic of 37.65 ± 0.40 and 38.37 ± 0.19 psu respectively (derived from average monthly values in the 1998-2007 period). It varied in intertidal zones, estuaries, close to under-sea freshwater springs and during rainy days in closed lagoons. At the same time, seawater temperatures showed some intra-seasonal differences (due to freshwater influence) in addition to strong seasonal variation. Salinity and temperature variations at investigated sites from Limski kanal to Dubrovnik (northern to southern Adriatic coast) showed freshwater influx which was also reflected in lower 13C values of mussel M. galloprovincialis shell carbonates. Based on the measured variables of seawater temperature, salinity and 13C, three groups of locations in the investigated area (25 locations) can be assigned: strong to medium freshwater influence (9 locations), minor freshwater influence (6 locations) and those of the marine environment (10 locations). Our results confirmed that mussel M. galloprovincialis sensitively responded to subtle changes in the environmental conditions. Results of the isotopic analyses presented in this study, support the hypothesis that the δ13C in mussel’s shell might be used as an indicator of environmental salinity stress. Therefore such investigations might represent an additional tool for waste water management and environmental protection.

Seawater salinity changes and thermal stress have been considered to be among the most important determinants of rocky intertidal zone organism distribution. The aim of present study was to determine the freshwater influx and evaluate the mussel Mytilus galloprovincialis shells as indicators of natural environmental conditions by comparing δ13C of shell carbonate layers with seawater salinity and tem- perature values measured over the 1998-2007 period. Our results show that the salinity was relatively constant in the open sea, with typical values of the northern and southern Adriatic being 37.65 ± 0.40 and 38.37 ± 0.19 respectively. It varied in intertidal zones, estuaries, locations close to under-sea freshwa- ter springs and during rainy days in closed lagoons on different locations along eastern Adriatic coast. Sa- linity and temperature variations at investigated sites showed freshwater influx which was also reflected in lower δ13C values of mussel M. galloprovincialis shell carbonates. Based on the measured parameters, three groups of locations in the investigated area (25 locations) can be defined: locations with strong to medium freshwater influence (9), with minor freshwater influence (6) and marine environment locations (10). Our results confirm that mussel M. galloprovincialis responds sensitively to subtle changes in the environmental conditions. Results of the isotopic analyses presented in this study support the hypothesis that the δ13C in mussel’s shell might be used as an indicator of environmental salinity conditions and hypo-osmotic stress. Therefore such investigations might represent an additional tool for waste water management and environmental protection., Promjena saliniteta i temperature morske vode u stjenovitom obalnom području ograničava i utječe na raspodjela morskih organizama. U području otvorenog mora salinitet je relativno stalan, a u istraživanom razdoblju (1998.-2007.) se kretao od 37,65 ± 0,40 za sjeverni i do 38,37 ± 0,19 za južni Jadran. Salinitet značajno koleba u zoni plime i oseke, na području ušća rijeka i podmorskim izvorima slatke vode te tijekom kišnih dana u zatvorenim lagunama. Zbog dotoka slatke vode temperatura mora na istraživanim postajama pokazuje unutar sezonske razlike, uz jaka sezonska kolebanja. Kolebanje vrijednosti saliniteta i temperature morske vode na području od Limskog kanala do Dubrovnika (sjeverni - južni Jadran) pod utjecajem dotoka slatke vode izravno su se odrazili i na vrijednosti δ13C u ljušturama dagnje Mytilus galloprovincialis. Vrijednosti δ13C iz 1998. godine su rangirane i korelirane s prosječnim vrijednostima temperature i saliniteta morske vode tijekom devetogodišnjeg istraživanog razdoblja (1998.-2007.). Na temelju sadržaja d13C u ljušturama, temperature i saliniteta morske vode istraživano područje (25 postaja) je grupirano u tri skupine: srednje do jak utjecaj slatke vode (9), manji utjecaj slatke vode (6) i lokacije morskog okoliša (10). Utvrđeno je da su ljušture školjkaša, odnosno vrijednosti δ13C, dobri pokazatelji kolebanja prirodnih čimbenika, naročito saliniteta, posredstvom dotoka slatkih voda. Dagnja M. galloprovincialis se može koristiti kao indikator kolebanja ekoloških čimbenika i možebitnih onečišćenja. Stoga navedena istraživanja predstavljaju dodatni koristan alat u biomonitoring programima, primjerice ispitivanju kvalitete okoliša, upravljanju otpadnim vodama i zaštiti okoliša.

Sponges are among the oldest multicellular animals and therefore very interesting from an evolutionary point of view. Current knowledge on their genome size and organization is contradictory. The aim of this study was an analysis of intron-size distribution in marine sponges. All poriferan protein-coding nuclear genomic sequences longer than 1 kilobase were retrieved from the National Center for Biotechnology Information (NCBI) database. Two sets of sequences were extracted from this initial group. The first set consisted of 18 genes with experimentally verified exon/intron structures. This set contained 15 genomic clones, in total 59 kilobases. The second set consisted of 14 unannotated Reniera sp. genomic stretches, in total 516 kilobases. Genscan software was used to predict the genes in both sets. Three groups of results were obtained, one experimentally verified and two predicted.The size distributions of exons and introns in the three groups of results were very similar. The distributions were highly skewed with median sizes of introns being 181, 184 and 204 nucleotides, and the expected difference in long introns making a much larger fraction in the Reniera sp. genes, with 30% of introns longer than 400 nucleotides compared to about 10% in the other two groups. Our results indicate that marine sponges have short introns.

Sponges are among the oldest multicellular animals and, as such, are very interesting from an evolutionary point of view. Current knowledge of genome size and organisation in sponges is contradictory. The aim of this study was a statistical exon/intron size analysis in sponges, in order to conclude about their genome organisation. The NCBI nucleotide sequence database was searched for Poriferan protein-coding genomic sequences longer than 1000 nucleotides, excluding mitochondrial genes and patents. The retreaved sequences were then used to search for corresponding cDNAs using the BLASTN algorithm. We finally obtained 18 cDNA sequences that matched to 15 genomic clones. Here we present a statistical exon/intron size analysis of all sponge genes with both the CDS and the genomic sequences deposited in the NCBI sequence database at the time of writing. The analysis was based on all complete exons contained within the CDSs and all introns between the start and stop codons. The average size of 78 exons, and 96 introns is 146 nt with a median of 119, and 259 nt with a median of 181 respectively. The regions between the start and stop codons of the analysed genes contain a total of 19143 coding and 24822 noncoding nucleotides making a very high "CDSs size" vs. "introns contained within" ratio of 0.77. Based on the positive correlation of intron size and genome size, we conclude that sponges have a compact genome.

Corresponding genomic and cDNA sequences of Suberites domuncula SdIrx-b Iroquois homeobox gene were presented. Sequence conservation and protein secondary structure prediction indicates only one of the two Iroquois homeodomains present in the gene is under selective pressure. SdIrx-b is probably an active gene but the second Iroquois homeodomain could be inactive.

Prikazana je građa i funkcija morskih spužava, osnovne karakteristike njihovih prilagodbi i interakcija sa drugim morskim organizmima.

Proteini koji sadrže homeodomenu transkripcijski su faktori vrlo važni za pravilan razvoj i održavanje homeostaze eukariotskih organizama. Do sada je kod spužve Suberites domuncula poznat Iroquois homeobox cDNA klon pod nazivom SdIrx-a, in situ lokalizacija ekspresije tog gena kao i utjecaj iona željeza i strujanja vode na njegovu ekspresiju. Pretragom genomske i cDNA biblioteke utvrđeni su: genomski slijed nukleotida gena SdIrx-a, genomski kao i cDNA slijed novog Iroquois gena nazvanog SdIrx-b, te treći genomski slijed nazvan SdIrx-α . Slijed SdIrx-b jedini je poznati homeobox gen koji kodira za dvije Iroquois homeodomene dok je SdIrx-α označen kao pseudogen. Zaključeno je da su slijedovi SdIrx-b i SdIrx-α najvjerojatnije nastali duplikacijom gena SdIrx-a. Na 5' strani gena SdIrx-a utvrđeno je postojanje gena TPPP/p25. Prema potencijalnim regulatornim elementima sadržanim u genomskom slijedu zaključeno je da je ekspresija gena SdIrx-a i TPPP/p25 vrlo vjerojatno povezana u obliku jedne dicistronske ili dvije preklapajuće mRNA. Na 5' strani gena SdIrx- α nalazi se gen COP1. Analiza mjesta izrezivanja introna iz gena TPPP/p25 i COP1 pokazuje vrlo visoku sačuvanost strukture ovih gena kod životinja.

AIF is a gene involved in the autoimmune response.The genomic locus of 28 kb in size was sequenced and here we report the current status of this work.

An understanding of cell-substratum adhesion to biomaterial surfaces is of value in promoting the attachment of connective tissue onto implant materials. In addition, adhesion of cells to implant surfaces is one of the major factors which determines the implant’s biocompatibility. The current definition of biocompatibility is that it refers to the ability of the material in question to perform with an appropriate host response in a specific situation (Williams, 1987). In certain circumstances on implant surfaces it is important to have adhesion of the soft tissues to prevent capsule formation and in other circumstances it may be desirable to have no adhesion, such as where tendons cross implants. Therefore measuring adhesion of appropriate cells and tissues to implant surfaces is very important for helping to screen new implant surface designs. Previously a semi-quantitative method of measuring the amount of cell adhesion was shown. This was based on general heavy metal staining, scanning electron microscopy (SEM) and image analysis of both the whole cell shape and the stained areas at the cell-substrate interface(Richards et al., 1996). Also Hunter et al., 1995, studied cell attachment using indirect immunofluorescent labelling of vinculin, quantifying the degree of cell attachment by determining the mean number of focal adhesions, containing vinculin, and determining the mean total area of the focal adhesions per cell. We now report an improved method for indirect immunocytochemical labelling of the integral protein vinculin within the cell focal adhesions for SEM viewing. The focal adhesions are either viewed, with the SEM, from above with the cells still on the substrate or from below, after the cells have been embedded in resin and the substrate removed. The advantages of this method over immunofluorescent labelling are: the possibility of high resolution viewing, there is no possibility of colour fading or autofluorescence problems - samples can be kept for years since gold particles do not disappear and also because of the particulate nature quantitation can be more accurate. There are two main advantages of this new method over the standard indirect immunolabelling for SEM viewing. 1. The literature (Beesley, 1993) suggests blocking (with the non-immune serum from the species that provides the 17 secondary antibody) to be performed before applying the primary antibody. We show that non-specific labelling is lower if this is performed just before the secondary antibody is applied. 2. Detergent extraction (using Triton X-100) is performed before the fixation. This is possible since it is known that Triton X-100 doesn’t extract vinculin from focal adhesions (Niederreiter et al., 1994). By this step most of the cell upper layers are removed allowing an easy visualisation of adhesion patterns in SEM with very low background labelling. The method will now be developed with image analysis techniques to allow a more accurate quantitative comparison of cell adhesion to different materials.

A good understanding of cell-substrate adhesion to biomaterial surfaces is important in promoting our understanding of the attachment of connective tissue onto implants. The adhesion of cells to implant surfaces is one of the major determinants of the implant's biocompatibility. Biocompatibility is currently defined as the ability of the material in question to perform with an appropriate response from the host in a specific situation. Normally on implant surfaces it is important to have adhesion of the soft tissues to the implant, so as to minimise capsule and "dead space" formation as well as the risk of infection. Under certain circumstances it may be desirable to have no adhesion of tissues, such as where tendons should slide across implant surfaces. Therefore, measuring the adhesion of appropriate cells and tissues to surfaces is very important for helping to screen new surface designs of materials for their various possible implant roles in the body. A semi-quantitative method of measuring the amount of cell adhesion based on estimating general heavy metal staining of cell adhesion sites by scanning electron microscopy and image analysis has been published. In the present study an improved method for indirect immunocytochemical labeling of the integral protein vinculin, within the cell focal adhesions, is used to confirm the correlation between the results of the semi-quantitative study specificity with vinculin containing adhesion sites.

Differences in adhesion of fibroblast and osteoblast cells on biocompatible materials with were quantified using immunogold labelling and scanning electron microscopy

Pri implantaciji metalnoga, keramičkog ili polimernog kirurškog pomagala u tijelo, zbivaju se na površini takvih naprava brojne reakcije. Upravo od toga kakve će reakcije neki materijal izazvati, ovisi njegova biokompatibilnost. Čimbenici koji određuju interakciju i adheziju živih stanica na biomaterijale od velikog su značaja za medicinu. Ovaj rad usmjeren je prema razvoju metode određivanja stupnja biokompatibilnosti materijala. Metoda imunocitokemijskog obilježavanja fokalnih kontakata stanica sa podlogom optimizirana je za obilježavanje proteina aktina, vinkulina, talina i integrina mišjih Balb/c fibroblasta za vizualizaciju rezultata epifluorescencijskim svjetlosnim mikroskopom. Optimizacija je uključivala: prilagođavanje fiksacije posebno za svako protutijelo, određivanje titra protutijela, uvođenja Tween 20, određivanja najboljih koncentracija BSA i Tritona X-100, izostavljanja određenih koraka i pronalaženja dobrog medija za uklapanje stanica.

Background: Leukocyte depletion before storage improves the quality of stored blood. We have studied the role of leukocyte depletion on blood viscosity. Study design and rnethodr: From 15 human CPDA-1 blood units specimen were taken before and after in-line filtration(Leuk0trap AlSystem). Blood viscosity (hematocrit 45%) was measured in Couette viscometer at 37°C at two different shear rates of 94.5 (primarily determined by red blood cell (RBC)deforrnability), and 0.1 s-' (primarily determined by RBC aggregation). Morphology of RBCs fixed with 1 % glutaraldehyde was assessed. The filtered and unfiltered samles were then stored at t4"C for 21 days, when blood viscosity and RBC morphology were again analyzed. Results: Directly after filtration, high shear viscosity (94.5 s ')was decreased from 4.7040.25 to 4.484-0.22 mPas (p=O.O002).whereas low shear viscosity remained unaffected (36.04-1 0.3 vs. 35.34-8.7 mPa s ; p=0.57). After storage, blood viscosity was increased at high and low shear rates for unfiltered and filtered samples (5.49+/-0.40 vs. 5.494-0.37 mPas, and 57.74-15.2 vs. 58.3+/-13.0 mPa s, respectively). An echinocytic shape transformation of RBCs was observed after storage, which correlated with the increase in viscosity. The RBC morphology was similar in filtered and unfiltered samples (morphological index: 1.6540.63 and 1.840.38, respectively). Conclurlonr: Leukocyte depletion initially improves high shear viscosity of CPDA-1 blood. This potential advantage is, however, completely lost after a storage period of 21 days. Improved metabolic conditions for RBC after prestorage leukodepletion as published in different studies, obviously did not result in a diminished shape transformation during storage.

Važnost pčela vrlo je velika, od oprašivanja biljaka među kojima su i ekonomski važne vrste, preko prehrambene i zdravstvene važnosti pčelinjih proizvoda, npr. meda, propolisa, peludi i matične mliječi i razne primjene pčelinjeg voska. Najbrojnija i najrasprostranjenija vrsta pčela je Apis mellifera, poznatija pod imenom medonosna pčela. One su ujedno i zadružni kukci s vrlo zanimljivim načinom života, podjelom uloga u zajednici i složenom međusobnom komunikacijom koja je primjer kompleksnosti počevši od jedinstvenog pčelinjeg plesa pa sve do izlaganja feromona. U ovom radu ukratko su prikazani najvažniji aspekti života pčela s posebnim naglaskom na ponašanje, odnosno međusobnu komunikaciju. Kako se obrasci ponašanja i stavovi djece pram okolini razvijaju od najmlađih nogu, izabrane su i dvije edukativne slikovnice za predškolsku djecu različite dobi koje su proanalizirane s ciljem mogućeg korištenja u predškolskim ustanovama. Pčele su iz više aspekata vrlo zanimljivi kukci i vrlo zahvalni kao podloga za osmišljavanje raznih edukativnih aktivnosti.

Uzimajući u obzir da obitelj predstavlja primarnu djetetovu okolinu od rođenja pa sve do adolescentske dobi postavlja se pitanje koja je njezina uloga u formiranju navika kod djece. U te navike ubrajamo i one prehrambene, koje bivaju sve češće propitkivane od strane zdravstvenih organizacija, zdravstvenih djelatnika ali i samih roditelja koji o njima žele znati više. Suvremeno doba donosi mnoštvo stručne i nestručne literature koja se usko tiče prehrane i prehrambenih navika te pruža mogućnost neprekidnog educiranja koje može pomoći u pravilnom formiranju prehrambenih navika u svakodnevnom životu. Prehrambene navike su izuzetno važne za psihofizičko stanje u svakoj dobi, a pogotovo u dječjoj kad je djetetu od strane obitelji nužna kvalitetna skrb i pažnja vezana za sve segmente djetetova rasta i razvoja. S ciljem utvrđivanja stavova roditelja koji su vezani za prehranu njihove djece predškolske dobi proveden je anketni upitnik koji je priložen u radu u kojem su ispitana 149 ispitanika. Pitanja u anketi odnosila su između ostalog na svakodnevne prehrambene navike kod djece. Anketom se ustanovilo da je majka ta koja vodi glavnu riječ u formiranju prehrambenih navika kod ispitanika. Considering that the family represents the child's primary environment from birth until adolescence, the question arises as to what role it plays in the formation of children's habits. These habits also include dietary habits, which are increasingly being questioned by health organizations, health professionals, and parents themselves who want to know more about them. Modern times bring a lot of professional and non-professional literature that closely concerns nutrition and eating habits and provides the possibility of continuous education that can help in the proper formation of eating habits in everyday life. Eating habits are extremely important for the psychophysical state at any age, and especially in the children's age, when the child needs quality care and attention from the family for all segments of the child's growth and development. With the aim of determining the attitudes of parents related to the nutrition of their preschool children, a survey questionnaire was conducted, which is attached to the paper, in which 149 respondents were questioned. The questions in the survey related, among other things, to the daily eating habits of children. The survey found that it is the mother who has the main say in the formation of the respondents' eating habits.

Plemenita periska (lat. Pinna nobilis) endem je Sredozemnog mora u kojem je i najveći školjkaš čija visina može dosegnuti i do 1 m. Najčešće je nalazimo okomito zabodenu na pjeskovito-muljevitom dnu, gdje naseljava livade morskih cvjetnica (Posidonia oceanica i Cymodocea nodosa). Ima važnu ekološku ulogu jer je filtratorski organizam te doprinosi prozirnosti vode. Njene ljušture služe kao tvrda površina koja biva kolonizirana od strane raznih epibionata. Još od 1992. godine nalazi se na popisu ugroženih vrsta, a njen se broj drastično smanjio nakon događaja Masovnog pomora koji je počeo u jesen 2016. godine. U ovom radu analizirano je 47 DNA sljedova s ciljem uvida u populacijsku genetiku vrste, a korišten je molekularni marker Citokrom oksidaza I, jedan od najraširenijih markera koji se koristi za evolucijske analize. Sljedovi su obrađeni u MEGA-X programskom paketu. Uzorkovanje je provedeno tijekom 2018. i 2019. godine na 7 lokacija duž hrvatske obale i otoka Jadranskog mora. Najvećim dijelom vidljivo je da je većina uzoraka genetski identična, odnosno među 47 analiziranih uzoraka pronađeno je 17 različitih haplotipova pri čemu je najbrojniji haplotip pronađen u čak 20 uzoraka i prisutan je na svih 7 uzorkovanih lokacija. Ukupno su detektirane 32 točkaste mutacije od čega samo jedna missense mutacija, dok su ostale tihe mutacije. Isto tako, usporedbom naših istraživanih skupina, pomoću BLAST-a, i ostalih populacija diljem Mediterana, čak 76% naših haplotipova, u punoj duljini od 553 nukleotida, identično je s već publiciranim sekvencama uzorkovanih diljem Mediterana. Pinna nobilis (lat. Pinna nobilis) is endemic to the Mediterranean Sea, where it is the largest bivalve whose height can reach up to 1 m. It is most often found stuck vertically on the sandy-mud bottom, where it inhabits meadows of Seagrass (Posidonia oceanica and Cymodocea nodosa). It has an important ecological role because it is a filter-feeder organism and contributes to water transparency. Its shells serve as a hard surface that is colonized by various epibionts. It has been on the list of endangered species since 1992, and its number decreased drastically after the Mass Mortality event started in the fall of 2016. In this work, 47 DNA sequences were analyzed to gain insight into the population genetics of the species, and the molecular marker Cytochrome Oxidase I was used, one of the most widespread marker used for evolutionary analyses. The sequences were processed in the MEGA-X software package. Sampling was carried out between 2018 and 2019 at 7 locations along the Croatian coast and the islands of the Adriatic Sea. For the most part, it is evident that most of the samples are genetically identical, that is, among the 47 analyzed samples, 17 different haplotypes were found, with the most numerous haplotype being found in 20 samples and present in all 7 sampled locations. A total of 32 point mutations were detected, of which only one was a missense mutation, while the rest were silent mutations. Likewise, by comparing our research groups, using BLAST, and other populations across the Mediterranean, as many as 76% of our haplotypes, in the full length of 553 nucleotides, are identical to already published sequences sampled across the Mediterranean.