Organisation of microtubules and spindle pole in the cells without centosomes

Najvažnija komponenta stanice odgovorna za sastavljanje mikrotubula je gama tubulin. On potiče dimerizaciju alfa i beta tubulina i tako nastanak mikrotubula. Lokaliziran je naročito na površini jezgre biljnih stanica koja je otkrivena kao glavna mjesta sastavljanja mikrotubula. Također, kortikalna membrana i fragmoplast su mjesta na kojima se mikrotubuli sastavljaju. Visoka dinamika sastavljanja i rastavljanja mikrotubula vrlo je bitna kod diobe stanice kada se mikrotubuli intenzivno premještaju. Proteini povezani sa mikrotubulima imaju važnu ulogu u njihovoj stabilizaciji te pomažu sastavljanje mikrotubula. Buduća istraživanja bi trebala razjasniti molekularnu funkciju proteina koji potiču izduživanje mikrotubula na acentrosomalnim polovima i unutar diobenog vretena. Također, kontrolni mehanizmi koji reguliraju funkciju mikrotubulskih organizacijskih centara su još nedovoljno otkriveni.The most improtant component of the cell, responsable for microtubule assembly is gammatubulin. Gamma-tubulin induce polimerisation of alpha and beta tubulin so as microtubule assembly. Its localized mostly on the nuclei surface, and this site is discovered as most active nucleating site of microtubules. Cortical membrane and phragmoplast are also sites of microtubule assembly. Highly dynamic assembly and dissasembly of microtubules is very important in mitosis when microtubules fast move in the cell. Proteins associated with microtubules are very important in stabilization of micortubules and for their assembly. Further researches should explain molecular function of proteins which induce polimerization of microtubules on acentrosomal poles and in mitotic spindle. Also, control points of regulation microtubule`s organization centers are still insufficient explained

Organisation of microtubules and spindle pole in the cells without centosomes

Najvažnija komponenta stanice odgovorna za sastavljanje mikrotubula je gama tubulin. On potiče dimerizaciju alfa i beta tubulina i tako nastanak mikrotubula. Lokaliziran je naročito na površini jezgre biljnih stanica koja je otkrivena kao glavna mjesta sastavljanja mikrotubula. Također, kortikalna membrana i fragmoplast su mjesta na kojima se mikrotubuli sastavljaju. Visoka dinamika sastavljanja i rastavljanja mikrotubula vrlo je bitna kod diobe stanice kada se mikrotubuli intenzivno premještaju. Proteini povezani sa mikrotubulima imaju važnu ulogu u njihovoj stabilizaciji te pomažu sastavljanje mikrotubula. Buduća istraživanja bi trebala razjasniti molekularnu funkciju proteina koji potiču izduživanje mikrotubula na acentrosomalnim polovima i unutar diobenog vretena. Također, kontrolni mehanizmi koji reguliraju funkciju mikrotubulskih organizacijskih centara su još nedovoljno otkriveni.The most improtant component of the cell, responsable for microtubule assembly is gammatubulin. Gamma-tubulin induce polimerisation of alpha and beta tubulin so as microtubule assembly. Its localized mostly on the nuclei surface, and this site is discovered as most active nucleating site of microtubules. Cortical membrane and phragmoplast are also sites of microtubule assembly. Highly dynamic assembly and dissasembly of microtubules is very important in mitosis when microtubules fast move in the cell. Proteins associated with microtubules are very important in stabilization of micortubules and for their assembly. Further researches should explain molecular function of proteins which induce polimerization of microtubules on acentrosomal poles and in mitotic spindle. Also, control points of regulation microtubule`s organization centers are still insufficient explained

MBD1 mediated enrichment of methylated DNA from cell lines deficient in S-adenosylhomocysteine hydrolase (AHCY)

Metilacija igra važnu ulogu u regulaciji bioloških procesa. Nepravilnosti u DNA metilaciji su uključene u nastanak različitih poremećaja zdravlja u ljudi. Nedostatnost S-adenozil homocistein hidrolaze (AHCY) je nova bolest, otkrivena u Hrvatskoj populaciji 2004. godine. Bolest rezultira u upečatljivom povišenju S-adenozil homocisteina (adoHcy) i S-adenozil metionina (adoMet) u krvi i različitim tkivima. AHCY uspješno uklanja adoHcy, nusprodukt transmetilacijskih reakcija i moćan inhibitor DNA metiltransferaza. Dakle, AHCY igra ključnu ulogu u regulaciji biološke metilacije. Cilj mog istraživanja bio je procijeniti promijene u DNA metilaciji nastale zbog nedostatnosti AHCY. Zato je DNA izolirana iz stanične linije ljudskog tumora jetre, HepG2, koja je utišana u AHCY ekspresiji korištenjem RNA interferencijske tehnologije. DNA metilacijski status je proučen na odabranim utisnutim lokusima te utvrđen bisulfitnim sekvenciranjem. Lokusi sa značajnim promjenama u metilacijskom statusu se mogu koristiti kao biomarkeri za detekciju bolesti.Methylation plays an important role in regulating biological processes. Abnormalities in DNA methylation are involved in the etiology of various human disorders. S-adenosylhomocysteine hydrolase (AHCY) deficiency is a novel human disease, discovered in Croatian population in 2004. The disease results in striking elevations of S-adenosylhomocysteine (adoHcy) and S-adenosylmethionine (adoMet) in blood and various tissues. AHCY efficiently removes adoHcy, the by-product of transmethylation reactions and potent DNA methyltransferase inhibitor. Thus, AHCY plays a critical role in regulation of biological methylation. The goal of my research was to evaluate AHCY deficiency derived changes in DNA methylation. To do so, DNA was isolated from human liver tumor cell line, HepG2, which has been silenced for AHCY expression using RNA interference technology. DNA methylation status was studied on certain imprinted loci and confirmed by bisulfite sequencing. Loci with significant changes in methylation status may serve as biomarkers for disease detection

MBD1 mediated enrichment of methylated DNA from cell lines deficient in S-adenosylhomocysteine hydrolase (AHCY)

Metilacija igra važnu ulogu u regulaciji bioloških procesa. Nepravilnosti u DNA metilaciji su uključene u nastanak različitih poremećaja zdravlja u ljudi. Nedostatnost S-adenozil homocistein hidrolaze (AHCY) je nova bolest, otkrivena u Hrvatskoj populaciji 2004. godine. Bolest rezultira u upečatljivom povišenju S-adenozil homocisteina (adoHcy) i S-adenozil metionina (adoMet) u krvi i različitim tkivima. AHCY uspješno uklanja adoHcy, nusprodukt transmetilacijskih reakcija i moćan inhibitor DNA metiltransferaza. Dakle, AHCY igra ključnu ulogu u regulaciji biološke metilacije. Cilj mog istraživanja bio je procijeniti promijene u DNA metilaciji nastale zbog nedostatnosti AHCY. Zato je DNA izolirana iz stanične linije ljudskog tumora jetre, HepG2, koja je utišana u AHCY ekspresiji korištenjem RNA interferencijske tehnologije. DNA metilacijski status je proučen na odabranim utisnutim lokusima te utvrđen bisulfitnim sekvenciranjem. Lokusi sa značajnim promjenama u metilacijskom statusu se mogu koristiti kao biomarkeri za detekciju bolesti.Methylation plays an important role in regulating biological processes. Abnormalities in DNA methylation are involved in the etiology of various human disorders. S-adenosylhomocysteine hydrolase (AHCY) deficiency is a novel human disease, discovered in Croatian population in 2004. The disease results in striking elevations of S-adenosylhomocysteine (adoHcy) and S-adenosylmethionine (adoMet) in blood and various tissues. AHCY efficiently removes adoHcy, the by-product of transmethylation reactions and potent DNA methyltransferase inhibitor. Thus, AHCY plays a critical role in regulation of biological methylation. The goal of my research was to evaluate AHCY deficiency derived changes in DNA methylation. To do so, DNA was isolated from human liver tumor cell line, HepG2, which has been silenced for AHCY expression using RNA interference technology. DNA methylation status was studied on certain imprinted loci and confirmed by bisulfite sequencing. Loci with significant changes in methylation status may serve as biomarkers for disease detection

Organisation of microtubules and spindle pole in the cells without centosomes

Najvažnija komponenta stanice odgovorna za sastavljanje mikrotubula je gama tubulin. On potiče dimerizaciju alfa i beta tubulina i tako nastanak mikrotubula. Lokaliziran je naročito na površini jezgre biljnih stanica koja je otkrivena kao glavna mjesta sastavljanja mikrotubula. Također, kortikalna membrana i fragmoplast su mjesta na kojima se mikrotubuli sastavljaju. Visoka dinamika sastavljanja i rastavljanja mikrotubula vrlo je bitna kod diobe stanice kada se mikrotubuli intenzivno premještaju. Proteini povezani sa mikrotubulima imaju važnu ulogu u njihovoj stabilizaciji te pomažu sastavljanje mikrotubula. Buduća istraživanja bi trebala razjasniti molekularnu funkciju proteina koji potiču izduživanje mikrotubula na acentrosomalnim polovima i unutar diobenog vretena. Također, kontrolni mehanizmi koji reguliraju funkciju mikrotubulskih organizacijskih centara su još nedovoljno otkriveni.The most improtant component of the cell, responsable for microtubule assembly is gammatubulin. Gamma-tubulin induce polimerisation of alpha and beta tubulin so as microtubule assembly. Its localized mostly on the nuclei surface, and this site is discovered as most active nucleating site of microtubules. Cortical membrane and phragmoplast are also sites of microtubule assembly. Highly dynamic assembly and dissasembly of microtubules is very important in mitosis when microtubules fast move in the cell. Proteins associated with microtubules are very important in stabilization of micortubules and for their assembly. Further researches should explain molecular function of proteins which induce polimerization of microtubules on acentrosomal poles and in mitotic spindle. Also, control points of regulation microtubule`s organization centers are still insufficient explained

Aberrations in <i>FGFR1</i>, <i>FGFR2</i>, and RIP5 Expression in Human Congenital Anomalies of the Kidney and Urinary Tract (CAKUT)

This study aimed to explore the spatio-temporal expression patterns of congenital anomalies of kidney and urinary tract (CAKUT) candidate genes, Fibroblast Growth Factor Receptor 1 (FGFR1), Fibroblast Growth Factor Receptor 2 (FGFR2) and Receptor-Interacting Protein Kinase 5 (RIP5), in human fetal kidney development (CTRL) and kidneys affected with CAKUT. Human fetal kidneys from the 22nd to 41st developmental week (duplex, hypoplastic, dysplastic, and controls) were stained with antibodies and analyzed by epifluorescence microscopy and RT−qPCR. The effect of CAKUT candidate genes on kidney nephrogenesis and function is confirmed by statistically significant variations in the spatio-temporal expression patterns of the investigated markers. The nuclear localization of FGFR1, elevated expression score of FGFR1 mRNA, the increased area percentage of FGFR1-positive cells in the kidney cortex, and the overall decrease in the expression after the peak at the 27th developmental week in dysplastic kidneys (DYS), suggest an altered expression pattern and protein function in response to CAKUT pathophysiology. The RT−qPCR analysis revealed a significantly higher FGFR2 mRNA expression score in the CAKUT kidneys compared to the CTRL. This increase could be due to the repair mechanism involving the downstream mediator, Extracellular Signal-Regulated Kinase 1/2 (ERK1/2). The expression of RIP5 during normal human kidney development was reduced temporarily, due to urine production and increased later since it undertakes additional functions in the maturation of the postnatal kidney and homeostasis, while the expression dynamics in CAKUT-affected kidneys exhibited a decrease in the percentage of RIP5-positive cells during the investigated developmental period. Our findings highlight the importance of FGFR1, FGFR2, and RIP5 as markers in normal and pathological kidney development