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PROTACs for targeted degradation of glycogen synthase kinase 3
No full textCiljana razgradnja proteina (TPD) predstavlja inovativan pristup u istraživanju i razvoju lijekova koji koristi prirodne stanične mehanizme za selektivno uklanjanje patoloških proteina. Posebno je značajna za ciljanje proteina na koje se klasičnim lijekovima malim molekulama ne može učinkovito djelovati. TPD ima velik potencijal u liječenju karcinoma, autoimunih i neurodegenerativnih bolesti.
Jedna od ključnih skupina molekula u TPD-u su kimere koje usmjeravaju ciljanu proteolizu, (engl. Proteolysis Targeting Chimeras, PROTAC-i). Ovaj diplomski rad opisuje sintezu i karakterizaciju PROTAC-a koji ciljaju enzim kinazu glikogen sintaze 3 (GSK3), ključnu kinazu u Alzheimerovoj bolesti.
Opisani PROTAC-i se sastoje od liganda za GSK3 (harmina), liganda za cereblon E3 ligazu (talidomida) i alkiltriazolske poveznice s tri, odnosno četiri ugljikova atoma u alkilnom lancu. U svrhu priprave PROTAC-a bilo je potrebno sintetizirati odgovarajuće prekursore: alkin harmina 2 i azide 3a-b. Alkin 2 dobiven je alkiliranjem harmina propargil-bromidom, dok su azidi 3a-b dobiveni diazo prijenosom iz odgovarajućih N-tert-butiloksikarbonil-monozaštićenih diamina i diazo donora 1H-imidazol-1-sulfonil azid hidroklorida 1. Alkin 2 i azidi 3a-b povezani su preko 1,2,3-triazolskog prstena u jednom reakcijskom koraku Huisgenovom 1,3-dipolarnom cikloadicijom, a nakon uklanjanja zaštitne tert-butiloksikarbonilne (Boc) iz spojeva 4a-b, dobiveni su amini 5a-b. Reakcijom kondenzacije amina 5a-b s talidomidskom kiselinom uz kondenzacijski reagens HATU sintetizirani su PROTAC-i 6a i 6b.
Spojevi su karakterizirani pomoću ¹H i ¹³C NMR, MS i FT-IR spektroskopije. Prema SwissADME analizi, spojevi imaju ograničenu oralnu bioraspoloživost, ne prelaze krvno-moždanu barijeru, djeluju kao inhibitori enzima citokrom P450 te su supstrati P-glikoproteina. Njihova biološka aktivnost bit će ispitana u daljnjim istraživanjima koja prelaze okvire ovog diplomskog rada.Targeted protein degradation (TPD) is an innovative approach in drug discovery and development that harnesses natural cellular mechanisms to selectively eliminate pathological proteins. It is especially important for targeting proteins that cannot be effectively modulated by conventional small-molecule drugs. TPD holds great potential in the treatment of cancer, autoimmune, and neurodegenerative diseases.
One of the major classes of molecules in TPD are proteolysis-targeting chimera (PROTAC). This master’s thesis focuses on the synthesis and characterization of PROTACs targeting glycogen synthase kinase 3 (GSK3), a key kinase implicated in Alzheimer’s disease.
The designed PROTACs consist of a GSK3 ligand (harmine), an E3 ligase ligand (thalidomide), and an alkyltriazole linker containing three or four carbon atoms in the alkyl chain. For the preparation of the target PROTACs, the appropriate precursors were synthesized: harmine alkyne 2 and azides 3a-b. Alkyne 2 was obtained by alkylating harmine with propargyl bromide, while azides 3a-b were synthesized by diazo transfer from the corresponding N-tert-butyloxycarbonyl-monoprotected diamines and the diazo donor 1H-imidazole-1-sulfonyl azide hydrochloride 1. Alkyne 2 and azides 3a-b were connected through a 1,2,3-triazole ring in a single reaction step using Huisgen 1,3-dipolar cycloaddition. After removal of the tert-butyloxycarbonyl (Boc) protecting group from compounds 4a-b, the corresponding amines 5a-b were obtained. Condensation of amines 5a-b with thalidomide acid using the coupling reagent HATU afforded PROTACs 6a and 6b.
The compounds were characterized using ¹H and ¹³C NMR, MS, and FT-IR spectroscopy. According to SwissADME predictions, the compounds exhibit limited oral bioavailability, do not cross the blood-brain barrier, act as cytochrome P450 inhibitors, and are substrates of P-glycoprotein. Their biological activity will be investigated in future studies beyond the scope of this thesis
Pharmacopoeial testing in the quality control of formulated dietary supplements containing slippery elm
No full textKora crvenog brijesta ima višestruku primjenu, a koristi se ponajviše kod upale grla, za probavne smetnje te pri iritaciji kože. Zbog nedostatka uspješnosti pri liječenju IBD konvencionalnim terapijama, kao i zbog nuspojava istih, ljudska se populacija sve češće okreće alternativnim oblicima terapije, osobito biljnim pripravcima lako dostupnim u obliku dodataka prehrani. Primjena dodataka prehrani za zdravstvene probleme sve je učestalija, a uz porast tržišta, problem predstavlja i činjenica kako dodaci prehrani ne prolaze kroz ispitivanja ekvivalentna onima koja se provode za lijekove dostupne na tržištu, kao i lakoća kojom se ti dodaci prehrani mogu nabaviti. U ovom radu provedena su ispitivanja varijabilnosti mase, raspadljivosti, gubitka sušenjem i bubrenja dozirnih jedinica dodataka prehrani na bazi crvenog brijesta namijenjenih liječenju IBD-a. Svi ispitivani uzorci bili su u obliku tvrdih kapsula. Cilj istraživanja bio je procijeniti usklađenost uzoraka s kriterijima Američke farmakopeje (USP-NF) kako bi se ocijenila njihova učinkovitost i sigurnost. Rezultati ispitivanja pokazali su da su svi uzorci, osim jednog, zadovoljili kriterije za varijabilnost mase. Dva uzorka nisu zadovoljila kriterij raspadljivosti, što upućuje na moguće smanjenje djelotvornosti. Gubitak sušenjem bio je unutar dopuštenih granica za sve uzorke, što sugerira prihvatljiv sadržaj vlage i hlapljivih tvari. Međutim, niti jedan uzorak nije zadovoljio kriterij bubrenja, što upućuje na smanjen sadržaj sluzi i smanjeni demulcentni učinak analiziranih dodataka prehrani. Dobiveni rezultati upućuju na potrebu za strožim nadzorom i kontrolom kvalitete dodataka prehrani, posebice onih koji se koriste u terapijske svrhe.The bark of the slippery elm has a wide range of applications, primarily used for sore throat relief, digestive issues, and skin irritation. Due to the limited effectiveness of conventional therapies for treating inflammatory bowel disease (IBD), as well as their associated side effects, people are increasingly turning to alternative treatment options, especially herbal remedies readily available as dietary supplements. The use of dietary supplements for health issues is on the rise, and alongside the growing market, a significant concern is that such products are not subjected to testing equivalent to that required for pharmaceuticals, and they are easily accessible without medical supervision. This study tested the mass variability, disintegration, loss on drying, and swelling of slippery elm-based dietary supplements intended for IBD treatment. All tested samples were in the form of hard capsules. This work aimed to assess the compliance of these samples with the United States Pharmacopeia (USP-NF) standards to evaluate their effectiveness and safety. The results showed that all but one sample met the USP-NF criteria for mass variability. Two samples failed to meet the disintegration criterion, indicating potentially reduced effectiveness. Loss on drying was within acceptable limits for all samples, suggesting an adequate moisture and volatile compound content. However, none of the samples met the USP-NF swelling criterion, indicating a lower mucilage content and reduced demulcent effect of the analyzed supplements. These findings highlight the need for stricter oversight and quality control of dietary supplements, especially those used for therapeutic purposes
Investigation of change in water content of swollen donepezil loaded microspheres at simulated nasal conditions
No full textOsjetilni učinci povezani s primjenom lijekova putem nosa važan su aspekt pri razvoju novih oblika lijekova. Prevelika viskoznost gelova ima negativan utjecaj na mehanizam mukocilijarnog čišćenja što može narušiti homeostazu nazalne sluznice i dišnog sustava. Isušivanje i stvaranje filma također su potencijalne mane gelirajućih sustava i mogu uzrokovati iritaciju nosne sluznice i smanjiti adherenciju pacijenata. In vitro ispitivanje gubitka vode iz gela i kroz gel prvi je korak u predviđanju osjetilnih učinaka nakon primjene in situ gelirajućih praškastih sustava za nazalnu primjenu.
Cilj ovog rada bio je ispitati promjenu sadržaja vode pri simuliranim uvjetima nosne šupljine u izbubrenim mikrosferama kao nosačima donepezila, indiciranog u liječenju blage do umjerene Alzheimerove bolesti. Metodom sušenja raspršivanjem pripravljene su mikrosfere koje sadrže: (i) donepezil i kitozan (DK), (ii) donepezil, kitozan i manitol (DKM) te (iii) donepezil, kitozan i laktozu (DKL), i kontrolnim fizičkim smjesama DK mikrosfera s manitolom (DK+M) i laktozom (DK+L). Mikrosfere su uspješno pripravljene sušenjem raspršivanjem, a iskorištenje procesa iznosilo je 14,5 – 60,1 %. Postignuta je uspješnost uklapanja donepezila u rasponu 91,4 – 93,2 % te je sadržaj lijeka iznosio 4,6 – 4,7 %. Bubrenjem svih pripravljenih praškastih uzoraka u umjetnom nosnom fluidu pripravljeni su gelovi za provedbu ispitivanja gubitka vode iz gela i kroz gel, pri simuliranim uvjetima nosne šupljine. Imerzijska ćelija kao nosač gela, pokazala se prikladnom za provedbu predmetnih ispitivanja. Konzistencija gelova koji su u svom sastavu imali laktozu bila je dostatna samo za orijentacijska mjerenja ispitivanih parametara gubitka vode. Za sve gelove inkubirane tijekom 8,5 h pri temperaturi od 34°C i 90% relativne vlage, uočena je linearna ovisnost kumulativnog gubitka mase vode o vremenu: (i) iz gela iznad praznog spremišnog odjeljka imerzijske ćelije, (ii) iz gela iznad spremišnog odjeljka imerzijske ćelije koji sadrži vodu, i (iii) iz spremišnog odjeljka koji sadrži vodu, kroz gel. Najveća brzina isparavanja vode iz gela iznad praznog spremišnog odjeljka imerzijske ćelije (44,3 mg/h) i iznad spremišnog odjeljka koji sadrži vodu (38,5 mg/h) zabilježena je za gel pripravljen bubrenjem DK mikrosfera. Isti gel karakteriziran je i najizraženijim okluzivnim učinkom u usporedbi s drugim ispitivanim gelovima, što upućuje na promjenu strukture konvencionalnog kitozanskog gela uslijed dehidracije. Najveća sposobnost zadržavanja vode DKM gela pripisana je prisustvu manitola te upućuje na najmanju mogućnost isušivanja i stvaranja polimernog filma na nosnoj sluznici te, posljedično, na najmanji rizik od izazivanja iritacije nosne sluznice. Time su DKM mikrosfere procijenjene kao praškasti sustav s najvećim potencijalom za neiritabilnu nazalnu primjenu.Sensory effects associated with nasal drug administration represent an important aspect in the development of innovative drug formulations. Excessive gel viscosity can negatively affect the mucociliary clearance mechanism, potentially disrupting the homeostasis of the nasal mucosa and the respiratory system. Dehydration and film formation are also potential drawbacks of gelling systems, which may cause nasal mucosa irritation and reduce patient adherence. In vitro evaluation of water loss from and through the gel is the first step in predicting the sensory effects following the application of in situ gelling nasal powders.
The aim of this study was to investigate changes in water content under simulated nasal cavity conditions in swollen microspheres used as carriers for donepezil, a drug indicated for the treatment of mild to moderate Alzheimer's disease. Spray dryng method was employed to prepare microspheres containing: (i) donepezil and chitosan (DK), (ii) donepezil, chitosan and mannitol (DKM), and (iii) donepezil, chitosan and lactose (DKL). Additionally, physical mixtures of DK microspheres with mannitol (DK+M) and lactose (DK+L) were used as control samples. The microspheres were successfully prepared with process yields ranging from 14.5% to 60.1%. The entrapment efficiency of donepezil ranged from 91.4% to 93.2%, and drug content ranged from 4.6% to 4.7%. Gell were obtained by swelling of the prepared powder samples in artificial nasal fluid, and subsequently used to evaluate water loss from and through the gels under simulated nasal cavity conditions. The immersion cell used as a gel holder proved to be suitable for conducting the planned investigations. The consistency of gels containing lactose was sufficient only for indicative measurements of water loss parameters. For all gels incubated for 8.5 hours at 34°C and 90% relative humidity, a linear relationship was observed between cumulative water mass loss and time: (i) from the gel above the empty reservoir compartment of the immersion cell, (ii) from the gel above the reservoir compartment containing water, and (iii) from the reservoir compartment containing water, through the gel. The highest evaporation rate from the gel above the empty reservoir compartment (44.3 mg/h) and above the water-containing reservoir (38.5 mg/h) was recorded for the gel prepared by swelling of DK microspheres. This gel also showed the most pronounced occlusive effect compared to other tested gels, indicating structural changes in the conventional chitosan gel due to dehydration. The greatest water retention capacity of the DKM gel was attributed to the presence of mannitol, suggesting the lowest potential for dehydration and polymer film formation at the nasal mucosa and, consequently, the lowest risk of mucosal irritation. Based on these findings, DKM microspheres were recognized as the powder with the greatest potential for non-irritating nasal administration
Synthesis and characterization of tetrahydropyrrolo[3,4- b]indole alkyne, a precursor of hybrid molecules with potential antimalarial activity
No full textMalarija je po život opasna zarazna bolest uzrokovana parazitima roda Plasmodium koji se prenose ubodom zaraženih
komaraca roda Anopheles. Unatoč značajnom napretku u kontroli i liječenju bolesti, malarija i dalje predstavlja
ozbiljnu javnozdravstvenu prijetnju, osobito u tropskim i supstropskim područjima. Rezistencija na postojeće lijekove
je u porastu, zbog čega je istraživanje novih načina liječenja malarije nužno. Ovaj rad obuhvaća sintezu i
karakterizaciju derivata tricikličkog prstena tetrahidropirolo[3,4-b]indola s propargilnom esterskom skupinom kao
ishodišne molekule u sintezi novih hibridnih molekula s potencijalnim antimalarijskim djelovanjem. U svrhu priprave
takvog derivata, modificirani su i kondenzirani indolski prsten i aminokiselina L-fenilalanin, pri čemu je pripravljen
N-tosil nitroindol u dva sintetska koraka. Propargilni ester aminokiseline je pripravljen u tri koraka (5), zatim je
amino- skupina spoja 5 prevedena u izocijanidnu skupinu u dva koraka (7). Izocijanidom modificiran propargilni ester
L-fenilalanina 7 korišten je u dearomatizaciji prethodno pripravljenog N-tosil nitroindola (2) pri čemu je sintetiziran
konačni produkt s karakterističnim tetrahidropirolo[3,4-b]indolskim prstenom (8). Strukture svih sintetiziranih spojeva
potvrđene su standardnim spektroskopskim metodama (1H i 13C NMR). Dobivena molekula će se u budućim
istraživanjima koristiti kao alkinski prekursor u klik-reakciji s azidima 4-amino-7-klorokinolinskih derivata, pri čemu
će nastati hibridi s triazolskom poveznicom potencijalnog antimalarijskog djelovanja.Malaria is a life-threatening infectious disease caused by Plasmodium parasites, transmitted through the bites of
infected Anopheles mosquitoes. Despite significant progress in disease control and treatment, malaria remains a
serious public health threat, particularly in tropical and subtropical regions. The increasing resistance to existing drugs
poses a major challenge, emphasizing the necessity for the synthesis of new antimalarial agents. This thesis
encompasses the synthesis and characterization of tetrahydropyrrolo[3,4-b]indole tricyclic ring derivatives with a
propargyl ester group as a starting molecule for the synthesis of new hybrid compounds with potential antimalarial
activity. To prepare such a derivative, modifications were made to the indole ring and the amino acid L-phenylalanine,
leading to the synthesis of N-tosyl nitroindole in two synthetic steps. The propargyl ester of the amino acid 5 was
synthesized in three steps, after which the amino group was converted into an isocyanide group in two steps. The
obtained isocyanide-modified propargyl ester of L-phenylalanine 7 was then used in the dearomatization of the
previously synthesized N-tosyl nitroindole (2), resulting in the synthesis of the final product featuring a characteristic
tetrahydropyrrolo[3,4-b]indole ring (8)
Optimization and validation of HPLC method for the simultaneous determination of prednisolone derivatives
No full textGlukokortikoidi su skupina lijekova koji se sve češće primjenjuju u suvremenoj medicinskoj praksi zbog svojih izraženih
protuupalnih i imunosupresivnih svojstava. Koriste se u liječenju širokog spektra upalnih bolesti, uključujući astmu,
upalne bolesti crijeva, alergije te različite bolesti oka. Ipak, njihova dugotrajna primjena povezana je s ozbiljnim
nuspojavama kao što su Cushingov sindrom, neurološke smetnje, glaukom i srčano zatajenje, što ističe potrebu za
razvojem pouzdanih analitičkih metoda za njihovo ispitivanje.
U ovom radu analizirani su su prednizolon fosfat i prednizolon acetat, a cilj je bio razviti i validirati HPLC metodu
sukladno ICH Q2(R2) smjernicama, te kvantificirati njihov sadržaj u prahu djelatne tvari i model uzorku. Primjenom
HPLC-DAD metode sa gradijentnom elucijom postignuto je učinkovito razdvajanje analita pri temperaturi od 50 °C i
valnoj duljini od 254 nm. Metoda je zadovoljila sve zahtjeve validacijskih parametara. Dobiveni rezultati pokazali su da
je sadržaj oba analita u ispitivanim uzorcima unutar propisanih granica prema važećim farmakopejskim standardima.Glucocorticoids are a group of drugs increasingly used in modern medical practice due to their pronounced antiinflammatory and immunosuppressive properties. They are used in the treatment of a wide range of inflammatory
diseases, including asthma, inflammatory bowel diseases, allergies, and various ocular conditions. However, their longterm use is associated with serious side effects such as Cushing's syndrome, neurological disorders, glaucoma, and heart
failure, highlighting the need for the development of reliable analytical methods for their examination.
In this study, prednisolone phosphate and prednisolone acetate were analyzed, with the aim of developing and validating
an HPLC method in accordance with ICH Q2(R2) guidelines and quantifying their content in the active pharmaceutical
ingredient powder and a model sample. By applying the HPLC-DAD method with gradient elution, effective separation
of the analytes was achieved at a temperature of 50°C and a wavelength of 254 nm. The method met all validation
parameter requirements. The obtained results showed that the content of both analytes in the tested samples was within
the prescribed limits according to current pharmacopoeial standards
Determination of iron and manganese in dietary supplements for the treatment of inflammatory bowel disease by atomic absorption spectrometry
No full textS obzirom na sve učestaliju pojavnost upalnih bolesti crijeva, raste i interes za dodatke prehrani
koji se koriste kao potporna terapija u njihovom liječenju. Budući da su dodaci prehrani u
Europskoj uniji regulirani kao hrana, a ne kao lijekovi, njihova je kontrola manje stroga, što ističe
važnost neovisnog ispitivanja njihove kvalitete. U ovom su radu analizirani dodaci prehrani koji
sadrže kurkumu, indijski tamjanovac, crni papar i justiciju. Ukupno je analizirano petnaest uzoraka
kupljenih u trgovinama zdrave hrane i putem internetske prodaje. Nakon mikrovalne digestije,
sadržaj željeza i mangana određen je metodom atomske apsorpcijske spektrometrije. Rezultati su
pokazali varijabilnost u koncentracijama ispitivanih metala među različitim pripravcima, pri čemu
su najviše vrijednosti zabilježene u uzorcima justicije i indijskog tamjanovca. Dobiveni rezultati
u skladu su s podacima iz dostupne literature, što potvrđuje pouzdanost korištene metode analize.
Analizirani dodaci prehrani pokazali su se kao vrijedan izvor željeza i mangana te potencijalno
mogu pridonijeti smanjenju nutritivnih deficita kod osoba s upalnim bolestima crijeva. Njihova
uloga u nadopunjavanju prehrane i očuvanju zdravlja dodatno potvrđuje potrebu za redovitom
kontrolom njihove kakvoće i sigurnosti.Given the increasing incidence of inflammatory bowel diseases (IBD), there is growing interest
in dietary supplements used as supportive therapy in their treatment. Since dietary supplements
are regulated as food in the European Union, rather than as medicines, their quality control is less
stringent, highlighting the importance of independent product testing. This study analyzed dietary
supplements containing turmeric, Indian frankincense, black pepper, and Green chiretta. A total
of fifteen samples were examined, purchased from health food stores and via online retailers.
Following microwave-assisted digestion, the content of iron and manganese was determined using
flame atomic absorption spectrometry. The results showed variability in the concentrations of the
analyzed trace metals among the different supplements, with the highest levels found in samples
containing Green chiretta and Indian frankincense. The obtained values were consistent with data
reported in the scientific literature, confirming the reliability of the analytical method used. The
analyzed dietary supplements proved to be valuable sources of iron and manganese and may help
reduce deficiencies of these essential micronutrients in individuals with IBD. Their role in
complementing the daily diet and maintaining health underlines the need for regular quality and
safety control of such products
Determination of cobalt, molybdenum and antimony in encapsulated dietary supplements using the GFAAS technique
No full textGlobalni porast prevalencije i incidencije upalnih bolesti crijeva potaknuo je razvoj i primjenu novih terapijskih pristupa, uključujući dodatke prehrani kao pomoćnu terapiju. Njihova sve šira dostupnost i kontinuirani rast potrošnje istaknuli su važnost sustavne procjene njihove kvalitete i sigurnosti, osobito u kontekstu dugotrajne primjene. S ciljem ispitivanja sigurnosti navedenih pripravaka, ovaj rad usmjeren je na određivanje koncentracija potencijalno toksičnih elementarnih onečišćenja antimona, molibdena i kobalta u kapsuliranim dodacima prehrani namijenjenima liječenju upalnih bolesti crijeva. Analiza je provedena primjenom grafitne atomske apsorpcijske spektrometrije (GFAAS), a rezultati su uspoređeni s propisanim vrijednostima dopuštenog dnevnog unosa (PDE) prema smjernicama ICH Q3D. Utvrđeno je da su sve izmjerene koncentracije elemenata ispod maksimalno dopuštenih granica, čime se potvrđuje sigurnost analiziranih pripravaka za oralnu primjenu. Nadalje, opažene su niže koncentracije elemenata u mekim kapsulama u odnosu na tvrde kapsule, što upućuje na moguće razlike u sastavu i tehnologiji formulacije.The global increase in the prevalence and incidence of inflammatory bowel disease (IBD) has prompted the development and use of new therapeutic strategies, including dietary supplements as adjunct treatment. The growing availability and widespread consumption of such products have highlighted the importance of systematically evaluating their quality and safety, particularly in prolonged use. This study aimed to assess the presence of potentially toxic elemental impurities, antimony, molybdenum, and cobalt in encapsulated dietary supplements intended for the management of IBD. The analysis was conducted using graphite furnace atomic absorption spectrometry (GFAAS), and the obtained concentrations were compared to the permitted daily exposure (PDE) limits defined by the ICH Q3D guidelines. All measured concentrations were below the established safety thresholds, confirming the safety of the analysed supplements for oral administration. Furthermore, lower concentrations of the investigated elements were observed in soft capsules compared to hard capsules, indicating potential differences in composition and formulation technology
Laboratory diagnostics of multiple myeloma
No full textMultipli mijelom aktivna je zloćudna bolest plazma stanica s izraženim oštećenjima drugih organa za razliku od stanja koje mu
prethode, monoklonske gamapatije neodređenog značaja i šuljajućeg mijeloma. Dolazi do stvaranja malignog klona plazma stanica koje sintetiziraju monoklonski imunoglobulin. Zbog njegovog nakupljanja, i potiskivanja funkcionalnih imunoglobulina, dolazi do najčešćih simptoma bolesti poput bolova u kostima, čestih infekcija, anemija, bubrežnog zatajenja i dr. Bolest je nepoznate etiologije, ali se pretpostavlja da joj pridonose genska predispozicija, izloženost zračenju i utjecaj okoliša. Druga je najčešća hematoonkološka bolest koja više zahvaća crnu nego bijelu rasu te više muškaraca od žena. Bolest se najčešće javlja između 65. i 72. godine života. Multipli mijelom neizlječiva je bolest, ali uz pomoć novih lijekova i autologne transplatacije matičnih stanica, postiže se puno dulje preživljenje i remisija bolesti. Laboratorijskim testovima dokazuju se monoklonski proteini, određuje se njihov tip te se kvantificiraju. Time se postavlja dijagnoza i prati uspješnost terapije. Elektroforeza serumskih proteina probirni je test za dokazivanje postojanja monoklonskog vrška. Određivanje slobodnih lakih lanaca u serumu probirni je test u kombinaciji s elektroforezom serumskih proteina i koristi se za praćenje bolesti te praćenje stanja nakon transplantacije koštane srži.
Imunofiksacijskom elektroforezom proteina klasicira se M-proteina. To je potvrdni test monoklonalnosti na temelju kojeg se može
donijeti diferencijalna dijagnoza mono-, bi- i triklonalnih gamapatija. Također, tim se testom provjerava relaps bolesti nakon
transplantacije koštane srži. Nalazi pacijenata s multiplim mijelomom često ukazuju na povišene koncentracije ukupnih proteina i
gama globulina, a koncentracija albumina je snižena. Uz to je sedimentacija eritrocita, kao biljeg upale i malignog procesa, ali i
viskoznosti krvi, ubrzana. Povišeni β-2-mikroglobulin loš je prognostički biljeg isto kao i hiperkalcijemija do koje dolazi zbog
osteolize u već uznapredovaloj bolesti. Cilj ovog diplomskog rada bio je prikazati mogućnosti biokemijskih metoda u dijagnostici
monoklonskih proteina kod sumnje na plazmastaničnu diskraziju. Koristeći se metodom kapilarne elektroforeze serumskih proteina monoklonski su vršci dokazani u 3 od 4 bolesnice, imunofiksacijom su potvrđeni tipovi monoklonskih proteina, a
imunoturbidimetrijom su kvantificirani. Navedene metode pomogle su u postavljanju dijagnoze i profiliranju fenotipa bolesti što je
preduvjet za daljnje liječenje.Multiple myeloma is an active malignant disease of plasma cells characterized by significant damage to other organs,
unlike its preceding conditions, monoclonal gammopathy of undetermined significance and smoldering myeloma.
The disease involves the formation of a malignant clone of plasma cells that synthesize monoclonal
immunoglobulin. Due to its accumulation and suppression of functional immunoglobulins, the most common
symptoms of the disease arise, such as bone pain, frequent infections, anemia, kidney failure, and others. The disease
has an unknown etiology, but it is believed that genetic predisposition, exposure to radiation, and environmental
factors contribute to it. It is the second most common hematologic malignancy, affecting Black people more than
White people, and men more than women. The disease most often occurs between the ages of 65 and 72. Multiple
myeloma is an incurable disease, but with the help of new drugs and autologous stem cell transplantation, much
longer survival and disease remission can be achieved. Laboratory tests detect monoclonal proteins, determine their
type, and quantify them. This establishes the diagnosis and monitors the effectiveness of therapy. Serum protein
electrophoresis is a screening test used to detect the presence of a monoclonal spike. The determination of free light
chains in serum is a screening test combined with serum protein electrophoresis, used for monitoring the disease
and post-bone marrow transplantation status. Immunofixation electrophoresis of proteins classifies the M-protein.
This is a confirmatory test for monoclonality, based on which a differential diagnosis of mono-, bi-, and triclonal
gammopathies can be made. Additionally, this test checks for relapse of the disease after bone marrow
transplantation. The laboratory findings of patients with multiple myeloma often indicate elevated levels of total
proteins and gamma globulins, while albumin concentration is reduced. Moreover, erythrocyte sedimentation rate
(as a marker of inflammation and malignant processes, but also blood viscosity) is accelerated. Elevated β-2-
microglobulin is a poor prognostic marker, as is hypercalcemia, which occurs due to osteolysis in advanced stages
of the disease. The aim of this thesis was to present the possibilities of biochemical methods in the diagnosis of
monoclonal proteins in suspected plasma cell dyscrasias. Using the method of capillary serum protein
electrophoresis, monoclonal spikes were detected in 3 out of 4 female patients, the types of monoclonal proteins
were confirmed by immunofixation, and they were quantified by immunoturbidimetry. These methods helped
establish the diagnosis and profile the disease phenotype, which is a prerequisite for further treatment
Traditional use of medicinal herbs in the Jastrebarsko area
No full textOvo je etnobotaničko istraživanje provedeno na području grada Jastrebarskog s naglaskom na prigradska
naselja Cvetković, Donji Desinec, Gornji Desinec, Prhoć i Prodin Dol, a u njemu je sudjelovalo 15 ispitanika
starije životne dobi (između 67 i 83 godine). Kriterij uključivanja u istraživanje bio je stjecanje znanja o
ljekovitom bilju putem usmene predaje. Ispitanici su sveukupno naveli 46 biljnih vrsta korištenih u liječenju,
a zabilježene su i njihove druge primjene. Najviše spomenutih biljnih vrsta pripadalo je porodicama
Asteraceae i Rosaceae, a prava kamilica, crna bazga, planinska čuvarkuća, malolisna lipa, obična kopriva i
vinova loza bile su ljekovite biljke od najvećeg značaja. Čajevi pripremljeni u obliku infuza bili su
najzastupljeniji ljekoviti pripravci, oralni put primjene bio je najučestaliji te su se listovi najčešće koristili u
njihovoj pripremi. Najveći broj biljnih vrsta primjenjivao se kod smetnji probavnog i dišnog sustava te kože.
Tradicionalna primjena biljaka u ljekovite svrhe zabilježena na području Jastrebarskog opravdana je i
potkrijepljena literaturnim podacima za većinu biljnih vrsta. Potrebno je provesti opsežnije i detaljnije
istraživanje u svrhu zapisivanja još raznolikije primjene biljaka navedenog područja s ciljem oplemenjivanja
i vrednovanja tradicije.This is an ethnobotanical study conducted in the area of the city of Jastrebarsko, with a focus on the suburban
settlements of Cvetković, Donji Desinec, Gornji Desinec, Prhoć, and Prodin Dol. The study involved 15
elderly participants (aged between 67 and 83). The inclusion criterion for the study was the acquisition of
knowledge about medicinal plants through oral tradition. The participants mentioned a total of 46 plant
species used in treatment, and their other uses were also recorded. Most of the mentioned plant species
belonged to the Asteraceae and Rosaceae families, with true chamomile, black elderberry, houseleek, smallleaved
linden, common nettle, and grapevine being the most significant medicinal plants. Teas prepared in
the form of infusions were the most common medicinal preparations, the oral route of administration was the
most frequent, and leaves were most often used in their preparation. The largest number of plant species was
used for digestive, respiratory, and skin disorders. The traditional use of plants for medicinal purposes
recorded in the Jastrebarsko area is justified and supported by literature data for most plant species. More
extensive and detailed research is needed to document even more diverse uses of plants from this area, with
the aim of enriching and valuing the tradition
Determination of metal impurities in dietary supplements for the treatment of inflammatory bowel diseases using the GFAAS technique
No full textPrimjena dodataka prehrani koji sadrže biljne sastavnice u stalnom je porastu, a zahtjevi za
kontinuiranu kontrolu sigurnosti istih ne postoje. Kurkuma, justicija, indijski tamjanovac i crni papar
sadrže biološki aktivne tvari koje pokazuju pozitivan učinak na tijek upalnih bolesti crijeva.
Navedene biljke iz tla akumuliraju, u svojim podzemnim i nadzemnim organima, teške metale koji
imaju negativan učinak na zdravlje ljudi. Određen je sadržaj kobalta, nikla, srebra i selena u 15
dodataka prehrani različita porijekla metodom atomske apsorpcijske spektrometrije, grafitnom
tehnikom. Kobalt i selen imaju važne uloge u organizmu, no u prekomjernim količinama ostvaruju
štetno djelovanje. Nikal i srebro štetni su već pri malim količinama. Analizi sadržaja metala
prethodila je mikrovalna digestija. Koncentracije analiziranih metala ispod su maksimalno
dozvoljenog dnevnog oralnog unosa za metalna onečišćenja koji navode ICH smjernice iz čega se
zaključuje da analizirani dodaci prehrani ne predstavljaju rizik za zdravlje. Pretraživanjem dostupne
stručne i znanstvene literature, rezultati rada uspoređeni su s prethodnim istraživanjima na istim
biljkama te nisu uočene statistički značajne razlike u sadržaju metala u biljkama.The use of dietary supplements containing herbal components is constantly increasing, yet the
requirements for continuous safety control of these supplements do not exist. Turmeric, Indian
frankincense, Green chiretta, and Black Pepper contain biologically active substances that show a
positive effect on the course of inflammatory bowel diseases. These plants accumulate heavy metals
from the soil in their underground and aboveground organs, which have a negative impact on human
health. The content of cobalt, nickel, silver, and selenium in 15 dietary supplements of various
origins was determined using atomic absorption spectrometry with graphite furnace technique.
Cobalt and selenium play essential roles in the body, but in excessive amounts, they have harmful
effects. Nickel and silver are harmful even in small amounts. Microwave digestion preceded the
analysis of metal content. The concentrations of the analyzed metals are below the permitted daily
oral intake for elemental impurities as stated by the ICH guidelines, leading to the conclusion that
the investigated dietary supplements do not pose a health risk. By searching the available
professional and scientific literature, the results of this work were compared with previous studies
on the same plants, and no statistically significant differences in metal content were observed in
plants