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Drugo izdanje priručnika daje sažeti pregled doprinosa molekularne biologije u području genetike i epigenetike, genomike, transkriptomike, proteomike, nutrigenomike i farmakogenomike, te prikaz modernih trendova u primjeni najnovijih dostignuća u kliničkim i znanstvenim laboratorijima.

U radu je prikazana kronologija životnog i znanstvenog puta slavnog hrvatskog genetičara, dr. Milislava Demereca.

Toliko toga treba uraditi u Europi i na Mediteranu redi očuvanja okoliša i zdravlja, na području zaštite ogromne biološke raznolikosti Mediterana, njegove kulturne i graditeljske baštine, te vrijedne prehrambene tradicije radi poboljšanja i unaprijeđivanja ljudskog zdravlja i zdravlja okoliša, što dobrim dijelom čeka i "European University Women" u toku 21. stoljeća. Mnoge pojedinosti o brojnim lošim primjerima iz cijeloga svijeta prikazane su u ovome radu da ih se podsjetimo i da ih upoznamo, da postanemo osjetljivi i svjesni kako ne bismo sami činili ili ponavljali iste greške. Očito je da postoji ogromni potencijal djelovanja za veliki intuitivni kapacitet mozga ljepšeg spola koji će sve to vjerovatno moći bolje napraviti od racionalnog mozga muškaraca. Vrijeme je sazrijelo da žene preuzmu mnogo aktivniju ulogu u oblikovanju budućnosti u ozračju mira i ljubavi, za zdravlje okoliša i zdravlje čovjeka u Mediteranu, u Europi i Svijetu. Upravo je jedna žena to i započela: dr. Gro Harlem Brundtland, bivša premijerka Norveške i sadašnja direktorica Svjetska Zdravstvene organizacije u Ženevi, potaknula je globalni proces koji je urodio formulacijom pojma Održivog Razvitka i koja je inspirirala Svijet da počne razmišljati o "Našoj zajedničkoj budućnosti".

Internacionalno bioetičko povjerenstvo UNESCO-a (IBC, International Bioethic Committee) predvodilo je višegodišnje napore u izradi Univerzalne deklaracije o ljudskom genomu i ljudskim pravima. Ovaj dokument sada je prihvaćen kao i Smjernice za njegovu djelotvornu provedbu u svim zemljama članicama. U smislu preporuka za implementaciju ove Deklaracije ona je prevedena i na hrvatski jezik i zaživjela je u svim oblicima dodiplomske i poslijediplomske nastave koja se odvija u obimu šesnaest različitih kolegija iz Katedre za medicinsku biologiju Medicinskog fakulteta Sveučilišta u Zagrebu.

Turizam kao najveća svjetska industrija sve se više preusmjerava u nove i raznolike oblike turističkih pothvata u malim razmjerima, za razliku od preživljenog masovnog turizma. Svima je njima zajedničko međutim unaprijeđenje zdravlja, dobrobit i rekreacija, sport i učenje u dokolici, interes za nove prostore i nove ljude, pa je prema tome turizam i najveća industrija psihičkog i fizičkog zdravlja. U svijetu sve većih i češćih epidemija i pandemija kojim industrijski proizvedena hrana ugrožava zdravlje i život čovjeka, vrijedno je prisjetiti se da je Mediteranska prehrambena kultura, kojom se tradicionalno hrani Dalmacija, Primorje i Istra, općeprepoznata u svjetskim dimenzijama kao najbolja prehrana za duži život i bolje zdravlje, kojom raspolaže zapadna civilizacija. Mediteranska prehrambena piramida trebala bi stoga postati jednom od najvažnijih turističkih atraktivnosti jadranskog turizma kao industrije zdravlja.

Background and purpose: 5-azacytidine (5azaC) is a demethylating agent and consequently a potential therapeutic for conditions where hypermethylation of genes is the cause of the disease. But prenatal exposure of rat and mice to a single dose of 5azaC resulted in fetal forelimb and hindlimb malformations as well as growth reduction. Pretreatment with acetylsalicylic acid (ASA) was able to diminish teratogenic effect of a single dose of 5azaC. In this work we tried to investigate ASA protective effect upon 5azaC teratogenicity if 5azaC was applied twice during pregnancy which caused more severe malformations in embryos. Material and Methods: 5azaC was dissolved in PBS and injected intraperitonealy at a concentration of 5 mg/kg body weight to pregnant rat females on 12th and again on 13th day of pregnancy. ASA was dissolved in PBS and administered every time 2 hr before 5azaC at a concentration of 75 mg/kg. Controls were treated with PBS on the same days of pregnancy. Animals were sacrificed in ether the day before birth. Fetuses and placentas were isolated from uteri, measured and weighted. For the differential staining of cartilage and bone fetuses were stained with alizarin red S and alcian blue. Results and Conclusions: All fetuses from 5azaC treated females showed both front and hind limb adactyly. If ASA was applied prior to 5azaC treatment, 46% of fetuses were normal with no malformations at all, while the rest showed only hind limb oligodactyly. In front limbs no malformations were discovered. Growth retardation caused by 5azaC was restored with ASA pretreatment in both embryos and placentas.

Filozofija i princip dizajna koji zovemo "PERMAKULTURA" utemeljio je u Australiji šumar dr. Bill Mollison i biolog David Holmgren, na kontinentu gdje su slatkovodni resursi još dragocjeniji od onih u Mediteranu. Ime je zapravo složenica od dvije riječi: PERMAnentna i agriKULTURA, i sugerira drugačiju filozofiju gospodarenja zemljištem i vodnim bogatstvom, te dizajniranja prostora ok njihovih kuća. Kasnije je taj pojam evoluirao u PERMAnentnu KULTURU, i stoga je to preteča pojma održivog razvitka. Najkraća definicija permakulture je priča o: 1. brizi za Zemlju, 2. skrbi za ljude, i 3. poštenim udjelima, koje smijemo uzimati za sebe od postojećeg prirodnog kapitala. Uloga voda ima središnje mjesto u permnakulturnom dizajnu i bitno odrađuje planiranje prostora i zona oko kuća i naselja. Čak i otpadne vode smatraju se "resursom" a ne problemom, i namjesto strategije odvodnje i dodatnog onečišćavanja mora i eutrofikacije priobalnih morskih ekosustava, podvrgavaju se recikliranju i pročišćavanju korjenskim sustavom umjetnih biljnih nasada, biotopa.

Mutageni su svuda oko nas i glavni put ulaza egzogenih štetnih faktora u tijelo je putem hrane koji uzrokuje mutagenezu i kancerogenezu. Tu su i endogeni faktori, koji potječu od slobodnih radikala kisika. Genetički modificirana hrana (GMO) kao hrana za čovjeka (food) i krmivo za stoku (feed) očituje se u problemeima diljem svijeta (StarLink kukuruz, zlatna riža na Filipinima, meksički grah). Zbog horizontalnog prijenosa gena - GMO je prekršten u LMO (l-living!)predstavlja ekološki rizik. UNESCO zahtijeva od znanosti da njihov napredak služi za dobrobit društva i čovječanstva, budući da je ponašanje znanstvenika pokazalo promašaje i obmane u tragedijama BSE-a, vCJD-a, koštanog brašna). Danas ima hrane koja nam je neprijatelj: mliječne masnoće, previše mesa, bijeli šećer, fast food.. No mi možemo prepoznati i hranu koja nam je prijatelj: zeleno, žuto i crveno voće i povrće, sa značajnom količinom antioksidansa, vitamina i polifenola. Ove tvari nas zaštićuju od mutagena i kancerogena u hrani i djeluju kao "presretači". Organska proizvodnja hrane i promocija takve prehrane jedina je razumna alternativa industrijskoj proizvodnji kakva danas prevladava u svijetu. To je naročito važno uvesti u Hrvatsku, kao mediteransku zemlju velikog turističkog potencijala. Mediteranska prehrambena piramida, zasnovana na svakodnevnom jedenju zdravih ugljikohidrata (kruh, žitarice, tjestenina, kukuruz), voća i povrća, umjerenom uzimanju mliječnih proizvoda i maslinovog ulja, te unosu proteina iz ribe, manje iz mesa, važna je komponenta hrvatskog turističkog razvitka.

Gastrulating rat embryo survives and differentiates in a organ-culture model in vitro. This model could be used as an alternative to in vivo approach for investigation of embryotoxic and teratogenic agents.

The report about Croatian SEMEP school activities has been carried out. The activities include diverse projects in SEMEP summer school on island Vis (2-9 July, 2000.) and preparing of SEMEP Seminar about Marine enviroment in Dubrovnik. Also there is a report about some new suggestions for further developing of SEMEP programme in Croatia orientated on Mediterranean Biodiversity Education.

Republic of Croatia participated in South Eastern Mediterranean Sea Enviromental Project (SEMEP) of UNESCO from its very inaguration in Athens, 1994. It is designed as student centered enviromental research programme and worksheets with set of modern instructions for students, teachers and coordinators. Such activities in Croatia were based on Adriatic Sea. At national level SEMEP included: Workshop on "The Role of Young People in Conservation of Nature and Culture" in Dubrovnik, The VIS'99 National SEMEP Summer School on th island of Vis, SEMEP Seminar in Split (1997) and in Bakar and Rovinj. It helped to establish better scientific communication with Split and SEMEP countries and to make new contacts. Also, the island Vis was considered as an great touristic destination. In a way of funny learning it can be achieved eco-tolerance and live for biological and landscape diversites.

Osma sjednica Internacionalnog bioetičkog povjerenstva UNESCO-a održana je od 12-14. rujna 2001. u Parizu, Francuska i oslovila brojne aktualne probleme i izazove: 1. ljudski genom -korištenje, pohrana i zaštita genomskih podataka, te 2. etika, genomika i intelektualno vlasništvo. Dio sjednice posvećen je i prezentaciji proteomike, kao nove istraživačke grane u post-genomskoj eri, kao i prikazu smjernica edukacije u bioetici. Savjetovanje o gore navedenim temema je nadalje provedeno u pitanjima načela iskazanih u Univerzalnoj deklaraciji o ljudskom genomu i ljudskim pravima.

As acetylsalicylic acid (ASA) diminished teratogenic effect of thalidomide we tried to investigate the effect of ASA in 5-azaC induced teratogenicity. Animals pretreated with ASA showed decreased fetal body weight loss and increased crown-rump length in comparison with those treated with 5-azaC. Number of almost of al malformations was reduced too.

Gastrulating rat embryo-proper contains three germ-layers with stem-cells which are able to differentiate in a serum-free organ-culture model and after cultivation retain potential to proceede to differentiate in transplants under the kidney capsule. Even organotypic structures such as tooth were found in such transplants.

Genetički modificirani organizmi (GMO) predstavljaju izvarendan doseg biomedicinskih znanosti i tehnologije rekombinantne DNA, koji pružaju velike potencijale u farmaceutskoj industriji, ali i veliku opasnost pri njihovom uvođenju u prehranu zbog očuvanja čovjekovog zdravlja i bioraznolikosti svijeta oko nas. Mutagene i kancerogene tvari danas su svuda oko nas i hranom najjednostavnije ulaze u naše tijelo. Također postoje veliki propusti u današnjoj industrijskoj proizvodnji hrane (koštano brašno) što je dovelo do nastanka i širenja Goveđe spongioformne encefalopatije (BSE) te za ljude pogubne varijantne Creutzfeldt-Jasobsove bolesti (vCJD). Ne zna se još u kakvoj su korelaciji te dvije bolesti. Prve žrtve BSE-a zabilježene su u Velikoj Britaniji 1984. godine, a do danas je utvrđeno 107 slučajeva vCJD-a u toj zemlji. Prehrambena industrija Velike Britanije doživljava i udar slinavke i šapa. Nedavno su zabilježene žrtve vCJD-a u Japanu, koje su se vjerojatno zarazile uvezenim mesom. Intezivna poljoprivreda danas ugrožava biološku raznolikost manipulirajući raznolikošću vrsta i njihovih gena, te povećanim iskorištavanjem prirodnih resursa i degradacijom okoliša. Zbog toga je usvojen model održivog razvitka na kojem se temelji Konvencija o biološkoj raznolikosti sa zadatkom očuvanja biodiverziteta i njegovo razumno korištenje te poštenu podjelu financijske dobiti od upotrebe genetskih resursa.

Sustainable Development is such a development which will enable full range of choices for prosperity and livehood of the present generation, and leave justified spectrum of choices for the new generations. It is a Hard Core Reality and Moral Imperative for us. Genetically modified organisms (GMO) represent at the moment the most outstanding, promising and controversial achievment of contemporary biomedical sciences and of recombinant DNA technology which offer great potential in pharmaceutical industry, but in agricultural application they are attracting ever stronger nutritional concerns and uncertainties about human health and health on biodiversity in Earth's ecosystems. These concerns about human health are proving real and acute problems and international community had to design obliging documents to conserve the endangered biodiversity (Convention of Biological Diversity) and control the spreading of living GMO (Cartagena Protocol on Biosafety). Enviromental mutagens and carcinogenes are all around us and the food is the major vector for their entry into our body. Today's food production is very intensive, with the use of many dangerous chemicals (PolyChlorinated Biphenyl, Meat Bone Meal etc.) and the philosophy of financial profits in food production lead to tragedy of Bovine Spongiform Encephalopathy (BSE) and variant Creutzfeldt-Jacobs Disease (vCDJ) which is probably just on the start. Biodiversity is disturbed also with the Habitat Loss, Invasion of non-native species, and with the Overuse of resources. It is essential to stop the Loss and retain the sustainable development, by regulating genetic resources and biotechnology, proving enviromental and nutritional education and raise public awareness.

Biološka raznolikost ili biodiverzitet (gr. bios, lat. diversitas) je pojam kojeg je predložio američki zoolog E.O. Wilson i koji obuhvaća širok raspon raznolikosti među živim organizmima, počev od temeljne genetske raznolikosti, raznolikosti među postojećim vrstama stvorenja, do raznolikosti među ekosustavima. Biodiverzitet je prema tome jedna priča o raznolikosti života, u bezbrojnom i nesagledivom mnoštvu i bogatstvu životnih oblika, koje je znanost biologija istom počela upoznavati i sagledavati njihove složene međuovisnosti.

Sedma sjednica Internacionalnog bioetičkog povjerenstva UNESCO-a održana je od 7-9. studenog 2000. u Quitu, Ecuador i oslovila brojne aktualne probleme i izazove poput: edukacije u bioetici, etički aspekti istraživanja embrionalnih matičnih stanica (izvještaj radne grupe IBC-a), solidarnost i internacionalna suradnja između razvijenih i zemalja u razvoju u pitanja istraživanja ljudskog genoma, ekonomskih aspekata istraživanja ljudskog genoma kao i prikaz stanja u znanstvenim istraživanjma o ljudskom genomu, neuroznanostima i starenju. Savjetovanje s udrugama bolesnika bilo je nadalje provedeno u pitanjima načela iskazanih u Univerzalnoj deklaraciji o ljudskom genomu i ljudskim pravima, kao i razvoja tehnologija o kojima je riječ.

Differentiation of superficial epithelia in rat embryo cultivated in vitro

Na poziv šefa Odsjeka za obrazovanje u znanostima i tehnologiji Edukacijskog sektora UNESCO-a da dam mišljenje, kritiku i korekcije novog pedagoškog priručnika za edukaciju u tim područjima za 21. stoljeće (Science and Technology Litaracy for All, 2000 +), proučen je dostavljeni materijal i mišljenje zajedno sa sugestijama i korekcijama odaslano u UNESCO-ovo sjedište.

The Greek Government has suggested to UNESCO Educational Sector the initiation of an international environmental education for students and teachers of vocational schools and high-schools entitled SEMEP: South Eastern Mediterranean Sea Environmental Project. This educational Project would like to offer a new pedagogical paradigm of research project based learning, student oriented holistic and interdisciplinary effort, which will address realprblems of local communities and enable outdoor student research activities and problem solving efforts

Državna uprava za zaštitu prirode i okoliša i Područni ured za zaštitu mora kao fokalna točka za djelovanje Mediteranskog programa tehničke pomoći u okolišu (METAP/CBU, Mediterranean Environmental Technical Assistance Programme/Capacity Building Unit) i Organizacija Ujedinjenih naroda za razvitak (UNDP, UN Development Programme) zajedno su poduzeli napore na izradi Plana održivog razvitka porječja rijeke Krke i Nacionalnog parka. Dvije nacionalne radionice su organiziane u Šibeniku uz sudjelovanje svih lokalnih čimbenika.

FGF negatively regulated growth of whole rat embryos cultivated in vitro but tissue-specifically preserved the potential for further growth.

Porječje rijeke Krke i obalno područje Šibensko-kninske županije predstavljaju dva područja sa specifičnim karakteristikama koje su međusobno povezane preko nekoliko prirodnih i socijalno-ekonomskih procesa. U tom smislu Plan održivog razvitka za slivno područje rijeke Krke i Nacionalni park Krka je razvijen i oblikovan Savjet za održivi razvitak ovog područja.

Neural retina from the eyes of 20-day-old Fisher rat fetuses was cultivated in a serum-free culture model at the air-liquid interface for 12 days or transplanted under the kidney capsule of adult rats for 15 days. During cultivation or after transplantation retina has lost its normal spatial relationships and rosettes were formed. Ultrastructural analysis confirmed survival of retinal cells even in the simplest protein-free medium in vitro. Photoreceptors, Mueller neuroglia, microglia and retinal plexiform layers were found.

Sastanak stručnjaka u obrazovanju o okolišu organiziran je od strane Vlade Galicije, pod pokroviteljstvom UNESCO-a u dvije faze: preparatorne (15-17. 11.) i središnje faze (20-24.11.2000.) u kojima je stotinjak eksperata iz pedesetak zemalja raspravljalo o Novim prijedlozima za djelovanje i korištenje obrazovanja o okolišu u pet tematskih područja: mirnog suživota na Zemlji, planinskim područjima i održivom turizmu, biološkoj raznolikosti i zaštićenim područjima, kompleksnosti okoliša i globalizacije, te korištenju obrzovanja o okolišu radi savladavanju siromaštva i gladi. Ovi su problemi odabrani od strane UNESCO-a jer oni predstavljaju zajedničke probleme na svim dijelovima svijeta.

DNA demethylation by 5-azacytidine in pregnant rat females impaired embryonic growth and caused malformations. Growth wasalso impaired in rat teratomas in vivo after application of 5-azacytidine.

Oko pedeset delegata ispred osamnaest Europskih biskupskih konferencija sudjelovalo je u radu konferencije. Oni su bili pozvani na sastanak od Savjeta Europskih biskupskih konferencija (CCEE, Consilium Conferentiarum Episcoporum Europae, St. Galen)

The results presented should be viewed as the solid basis for more detailed studies of the imapact of developmental and teratogenic factors on postimplantation mammalian development in chemically defined culture conditions which have not yet been developed for whole-rat-embryo cultures.

Načela medicinske genetike

Predložen je program obrazovanja o okolišu u Jadramsko-jonskom bazenu

AIM OF THE STUDY : 5-azacytidine is a known demethylation agent which can be incorporated into DNA instead of cytosine and prevent its methylation. Therefore, its incorporation into DNA can change the established pattern of gene activation and influence the differentiation. We tried to investigate the influence of demethylation on various tissues in young rats by i.p. application of 5-azacytidine. METHODS: 13 day-old rats were i.p. injected with 5 and 10 mg/kg 5-azacytidine three times during one week. Rats were sacrified after four weeks and testicles fixed in 4% glutaraldehyde, postfixed in 1% OsO4 and embedded in Durcopan. Semi-thin sections were analysed with light microscopy and transmission electron microscopy was done on ultra-thin sections. RESULTS: 13 day-old rats showed impaired testicular development with disturbed spermatogenesis in both sets of experiments. The development was more disturbed with the higher dose of 5-azacytidine. In the affected testicles, some seminiferous tubules were deprived of spermatogenic cells. A number of tubules showed desquamation and disorganisation of seminiferous epithelium. Sertoli cells displayed huge vacuoles and incresed presence of lipid droplets in their cytoplasm. Ultrastructural investigation of mentioned vacuoles demonstrated their low-electron density. CONCLUSION: In the present study it was shown that the process of methylation is crucial for spermatogenesis as well as testicular development in the postnatal period of the rat. It is speculated that 5-azacytidine has a stage-specific effect on the development of spermatogenic cells in the rat.

Porječje rijeke Krke i obalno područje Šibensko-kninske županije predstavljaju dva područja sa specifičnim karakteristikama koje su međusobno povezane preko nekoliko prirodnih i socijalno-ekonomskih procesa. U tom smislu Plan održivog razvitka za slivno područje rijeke Krke i Nacionalni park Krka je razvijen i oblikovan Savjet za održivi razvitak ovog područja.

In the organ culture of gastrulating rat embryos without extraembryonic parts it is possible to investigate various signals required for normal or impaired development (Bulić-Jakuš et al., 1990 ; Škreb et al., 1993 ; Vlahović et al., 1993 ; Strahinić et al., 1996). The aim of the study presented here was to evaluate the degree of neural tissue differentiation in cultivated rat embryos with electron microscopy and immunohistochemistry. Fisher rat embryos 9, 5 days old were microsurgically isolated and cultivated for 14 days on a metal grid in Eagle's Minimal Essential Medium with 50% rat serum. Resulting teratoma-like eksplants were fixed in 4% glutaraldehyde, postfixed in 1% OsO4, embedded in Durcopan, sectioned for transmission electron microscopy (TEM) and analysed with Zeiss 902 A microscope. Some of the explants were fixed in St. Marie's solution, embedded in paraffin, serially sectioned and processed for immunohistochemistry. Primary antibody against Proliferating Cell Nuclear Antigen ( anti-PCNA, DAKO) and LSAB2 peroxidase kit (DAKO) for detection of the antigene-antibody complex were used. After two-weeks of culture, among differentiated tissues in embryonic explants, features typical for neural tissue were discovered with TEM : neuropil and myelinization in the brain. A strong intranuclear PCNA signal was also discovered in some cells which indicated that they were still proliferating. It was concluded that after cultivation of gastrulating rat embryos, terminal differentiation of brain cells has occurred although some cells were still retained in the cycling compartement.

U sklopu treće faze Sredozemnog programa tehničke pomoći za okoliš (engl. Mediterranean Technical Assistance Programme, METAP III) i uz njihovu financijsku potporu, u organizaciji Državne uprave za zaštitu prirode i okoliša (DUZPO) - Područne jedinice za zaštitu mora, kao nacionalnog središnjeg mjesta za taj Program, u Šibeniku je održana Nacionalna radionca Krka radi izrade Plana održivog razvoja i investicijski program za rijeku Krku i Nacionalni park Krka.

Znanstveni poslijediplomski studij temeljnih medicinskih znanosti ima za cilj postizanje akademskog stupnja magistra i doktora znanosti. Tijekom tog studija postdiplomandi uče znanstvenu metodologiju uz niz izbornih predmeta iz oblasti medicine čime se osposobljavaju za daljnji samostalni znanstveni rad.

Gastrulating rat embryo, devoid of extraembryonic parts, when cultivated for two weeks in vitro, gives rise to various differentiated tissues in serum-supplemented and even in serum-free chemically defined culture media. This model was used before to investigate various developmental signals (Bulić-Jakuš et al. International Journal of Developmental Biology 34, 275-279, 1990 ; Vlahović, M. et al. Periodicum Biologorum 96, 65-66, 1993). The aim of the present study was to compare the acivity of teratogen 5-azacytidine (5azaC) in serum-free and serum-supplemented culture conditions. Also a more precise investigation of developmental parameters (growth and differentiation) was done. Fisher rat egg-cylinders 9, 5 days old were microsurgically isolated and their embryonic parts cultivated for two weks in protein-free Eagle's Minimal Essential Medium (MEM) as well as in MEM enriched with 50 ľg/ml transferrin or 50% rat serum. Light microscopy, transmission electron microscopy and immunohistochemical detection of Proliferating Cell Nuclear Antigen (PCNA) showed that terminal differentiation of some tissues (e.g. epidermis, neural tissue) has occured, but also that some cells were still retained in the cycling compartement. In serum-free media one dose of 5azaC was able to impair survival, growth and differentiation of embryos, while in serum-supplemented medium only repeated application of 5-azacytidine impaired growth. Moreover, in serum-supplemented medium differentiation with 5azaC was even better than in controls, probably because of its DNA demethylating property which causes gene activation. The above results demonstrated that in chemically defined media 5azaC exhibited its effect promptly and in concordance with its known embryotoxic activity.

Methylation is an important mechanism for regulation of gene expression during vertebrate development. 5-azacytidine is usually used as an experimental tool for demethylation. We have used 5-azacytidine ( 5mg/kg body weight) in different stages of rat development. It was applied in vivo by i.p. injection to pregnant Fischer rat females from day 1 to day 16 of pregnancy. Embryos were isolated and examined the day before birth. After the application of 5-azacytidine on the first three days no changes whatsoever were found. On the fourth day of pregnancy 5-azacytidine caused reduction of growth which was present in all living fetuses found after that day. From day 5 till day 11 of application no living fetuses were found. On day twelve 24% of fetuses survived, but forelimb and hindlimb malformations were present. On the day 13, 5-azacytidine did not interfere with survival, but malformations were still present. From day 14 on, no gross external malformations were found. Placentas were also influenced by 5-azacytidine. They were significantly smaller and histological evaluation showed that labyrinthine part was severely reduced, but on the contrary trophoblast giant cells were more abundant than in controls. These results demonstrate that 5-azacytidine did not affect only the developing embryo, but as well impaired the placenta in a specific manner.

The KRKA Project was chosen as a Croatian priority as it represents a new approach toward sustainable development planning at the local level for the Šibenik-Knin County. Republic of Croatia was invited to present this project as the case study, as one among the eight Mediterranean countries which has started this process.

Sredozemno more doživljava povremene ekološke krize na raznim mjestima i s različitim stupnjem intenziteta. Veliki pomori dupina uzrokovani su intoksikacijom ovih morskih sisavaca polikloriranim bifenilima, čime je njihova imunološka obrana znatno oslabljena, pa smo svjedočili morskom obliku stečene imunodeficijencije zbog koje su tisuće dupina u zapadnom Sredozemlju uginuli kao žrtve pomora od neposrednog uzročnika smrti: epidemije morbili virusa. Zelena alga Caulerpa taxifolia, nakon što je doživjela značajne genetičke promjene u akvarijima Europe, pobjegla je iz akvarija u Monaku i udomaćila se u obalnom području Sredozemnog mora Francuske, Španjolske, Italije, a sve na uštrb polja morske cvjetnice, trave Posidonia oceanica . Ova podmorska polja trave, nekada su bila mjesta za mrijest riba i bogatstvo svekolikog života u obalnom području Sredozemlja. Danas su ta prirodna mrijestilišta zamijenjena poljima tropske zelene alge, koja živi kao monokultura i nemilosrdno prodire kako na zapad, tako i na istok. U Jadranu imamo zlu sreću njenih nalaza već na tri mjesta. Znači li to kraj i našim podmorskim poljima posidonije, našim mrijestilištima, ribama, raznolikosti bioloških vrsta i podmorskih krajobraza Jadrana?

Croatian Strategy for Environmental Education in Church and School is the action in progress, a process during the last decade of this millenium: the Decade of the Environment: 1990-2000.

We were investigating whether mammalian epidermis which differentiates in embryo culture in vitro retains the ability to give rise to skin appendages in a richer environment in vivo. Epidermis differentiated only after 14 days of culture and was mostly terminally differentiated. Only after subsequent grafting of cultured explants under the kidney capsule skin appendages (hair and sebaceous glands) differentiated. So far no difference in their incidence related to previous medium supplementation or culture period was discovered. It seems that developmental potential of epidermis to give rise to skin appendages was not harmed by previous culture in serum-free and protein-free medium.

Velebit Mountain biosphere reserve as the only one in Croatia was registered with UNESCO Ecological sector and the MAB Programme for twenty years. Statutary Framework of the World Network of Biosphere Reserves defines minimal criteria for nature conservation which would be seriously offended with potential construction of the coal-powered energy plant. Scientific, expert and public campaign staged by mountaineering union helped prevent such a deleterious construction at the foot the biosphere reserve.

The Sustainable Development Plan and Investment Programme for the National Park Krka and River Krka project was selected by the State Directorate for the Protection of Nature and Enviroment and the Goverment of Croatia, under METAP III. The project is considered as a capacity building initiative for the implementation of the sustainable development at the local level in the Šibenik-Knin County.

Skup tekstova iz područja biologije reprodukcije koji se nudi kao čitanka (engl. reader) studentima velikog integriranog predmeta (VIP) Majka i dijete u dvanaestom semestru studija medicine.

The new UNESCO environmental education project SEMEP (South Eastern Mediterranean Sea Environmental Project) addresses high school students and teachers to initiate an interdisciplinary and holistic education effort. The first research based learning effort was concentrated on the thematic issue on the interrelationship of Man and the Sea. Students were encouraged to do local research on beach area and the two man-made structures by the sea. Many students and teachers from the whole of the Adriatic Coast have taken the active part in these studies.The first Croatian SEMEP Summer School was organized in Rovinj and the first International Summer School in the American Farm School in Thessaloniki, Greece

Until now 'c-series' polysialogangliosides were known to exist in human brain only during development and in some pathological conditions like Alzheimer's disease. Using thin-layer chromatography (TLC) and immunostainng with Q211 antibody (TLC-overlay techniwue) we have analysed 'c-series gangliosides in four human cerebella (age 20, 47, 52 and 54 years). Four distinct ganglioside bands, most probably corresponding to GT1c, GQ1c, GP1c and GH1c were found to exist in the analysed brain, which is convincing demonstration of the existence of c-series" gangliosides in normal adult human brain. Immunohistochemical analysis was performed to locate polysialogangliosides in the analysed tissu. Q211 antibody was found to bind specifically to a single subpopulation of neurons in the molecular layer of adult cerebellum. According to their position and morphology these cells correspond to stellate neurons.

Svrha ovog preglednog rad je da pokaže neurotoksični i neuroteratogeni učinak metil-žive, otrova koji se nakuplja u morskim hranidbenim lancima i konačno u ribi. Pregled prikazuje kronologiju događaja i reakciju društva u dobro dokumentiranom trovanju organskom živom porijeklom iz zaljeva Minamata i Niigati, kao i u Iraku. Prikazuje postojeće spoznaje o nekim toksičnim i teratogenim mehanizmima metil-žive, gdje je mozak, naročito fetalni mozak, najugroženiji organ, te s tim u vezi objašnjava nastala neurološka oštećenja i promjene ponašanja.

Theodoros Christides, predsjednik Grčke organizacije za vokacijsko obrazovanje (O.E.E.K) u okviru UNESCO-ovog projekta okoliša Jugoistočnog Sredozemnog mora konstatira: Mi smo kao prosvjetni radnici pozvani da iskoristimo svoje jedino oružje koje nam preostaje, a to je snaga prosvjete, kako bismo stvorili nove ljude, s novim vizijama i sa željom da nametnu nove odnose i učinkovita rješenja za zajedničke probleme našeg okoliša. Mi prosvjetni radnici, moramo surađivati i razviti zajedničke programe koji su usmjereni prema istome cilju.