Comparative analysis of expression of angiogenic factors and CD44 gene in human glioma and neuroblastoma cell lines in vitro

Angiogenesis is essential for tumor growth and relies on the production of angiogenic factors. By comparative analysis using RT-PCR method of angiogenic growth factors: VEGF, bFGF, PDGF-A, angiogenin- 1 and IL-8 we established the level of expression of these genes necessary for angiogenesis in glioma and neuroblastoma cell lines. Our analyses were also extended to CD44 gene, which plays an important role in cascade of metastasis and progression of brain tumors. Significant differences in the level of gene expression of angiogenic factors and CD44 gene between the two cell lines observed throughout this study can be used as a prognostic marker for predicting clinical outcome in human brain tumors at the time of the initial staging.Angiogeneza je neophodna za rast tumora i zahteva proizvodnju angiogenih trofičkih faktora koji učestvuju u tumorogenezi. Uporednom analizom angiogenih trofičkih faktora: VEGF, bFGF, PDGF-A, angiogenina-1 i IL-8 pomoću metode RT-PCR utvrdili smo nivo ekspresije ovih gena uključenih u proces angiogeneze u ćelijskim linijama glioma i neuroblastoma. Takođe smo proširili analize i na CD44 gen koji igra važnu ulogu u kaskadi nastanka i progresiji metastaza tumora mozga. Dobijeni rezultati ukazuju na značajnu razliku u nivou genske ekspresije angiogenih faktora i CD44 gena u ove dve ćelijske linije čije se poreklo razlikuje ne samo po nastanku već i po mestu rasejavnja metastaza. Rezultati bi mogli da posluže kao prognostički faktor u prekliničkim i kliničkim istraživanjima tumora mozga od inicijalnih do terminalnih stupnjeva nastanka i terapije

6-hydroxydopamine lesions of the striatum lead to the alterations of dopamine receptor mrna in parkinsonian rats

The effects of four-site intrastriatal 6-hydroxydopamine (6-OHDA) lesions were examined in adult male rats. Five days after the lesions the animals were checked for specific rotational behavior induced by middle dose of amphetamine and the results confirmed the effectiveness of the lesions. The RNAs from the striatum were isolated at different time points after the lesion, and the RT-PCR analyse were performed for the D1 and D2 receptor mRNA. The results show a decline in the D2 receptor mRNA level (40%) at 6 h and 24 h points while this change was not observed seven days after the lesion. In contrast, no statistically significant changes in the level of the D1 receptor mRNA after the lesion at any time point were found.Ispitivani su efekti četiri ubodne 6-hidroksidopaminske (6-OHDA) lezije striatuma kod odraslih mužjaka pacova. Pet dana nakon lezije, životinje su testirane na specifično rotaciono ponaš anje pod uticajem srednje doze amfetamina i rezultati su potvrdili efikasnost lezije. RNK iz striatuma su izolovane u različitim vremenskim tačkama nakon lezije i urađena je RT-PCR analiza iRNK za D1 i D2 dopaminske receptore. Rezultati pokazuju smanjivanje nivoa iRNK za D2 receptor (40%) 6 h i 24 h nakon lezije, dok sedam dana nakon lezije nema promena. Za razliku od ovih rezultata, u nivou iRNK za D1 receptor ne postoje statistički značajne razlike u bilo kojoj vremenskoj tački

Changes in expression of GFAP, ApoE and APP mRNA and protein levels in the adult rat brain following cortical injury

The recovery period following cortical injury (CI) is characterized by a dynamic and highly complex interplay between beneficial and detrimental events. The aim of this study was to examine the expressions of Glial Fibrillary Acidic Protein (GFAP), Apolipoprotein E (ApoE) and Amyloid Precursor Protein (APP), all of which are involved in brain plasticity and neurodegeneration. Our results reveal that CI strongly influenced GFAP, ApoE and APP mRNA expression, as well as GFAP and ApoE protein expression. Considering the pivotal role of these proteins in the brain, the obtained results point to their potential contribution in neurodegeneration and consequent Alzheimer's disease development.Projekat ministarstva br. 17305

Comparative analysis of expression of angiogenic factors and CD44 gene in human glioma and neuroblastoma cell lines in vitro

Angiogenesis is essential for tumor growth and relies on the production of angiogenic factors. By comparative analysis using RT-PCR method of angiogenic growth factors: VEGF, bFGF, PDGF-A, angiogenin- 1 and IL-8 we established the level of expression of these genes necessary for angiogenesis in glioma and neuroblastoma cell lines. Our analyses were also extended to CD44 gene, which plays an important role in cascade of metastasis and progression of brain tumors. Significant differences in the level of gene expression of angiogenic factors and CD44 gene between the two cell lines observed throughout this study can be used as a prognostic marker for predicting clinical outcome in human brain tumors at the time of the initial staging.Angiogeneza je neophodna za rast tumora i zahteva proizvodnju angiogenih trofičkih faktora koji učestvuju u tumorogenezi. Uporednom analizom angiogenih trofičkih faktora: VEGF, bFGF, PDGF-A, angiogenina-1 i IL-8 pomoću metode RT-PCR utvrdili smo nivo ekspresije ovih gena uključenih u proces angiogeneze u ćelijskim linijama glioma i neuroblastoma. Takođe smo proširili analize i na CD44 gen koji igra važnu ulogu u kaskadi nastanka i progresiji metastaza tumora mozga. Dobijeni rezultati ukazuju na značajnu razliku u nivou genske ekspresije angiogenih faktora i CD44 gena u ove dve ćelijske linije čije se poreklo razlikuje ne samo po nastanku već i po mestu rasejavnja metastaza. Rezultati bi mogli da posluže kao prognostički faktor u prekliničkim i kliničkim istraživanjima tumora mozga od inicijalnih do terminalnih stupnjeva nastanka i terapije

Apoptotic clearance in rabbits with experimental pulmonary emphysema

In order to better understand pathogenesis of pulmonary emphysema, the model of experimentally induced pulmonary emphysema in Chinchilla rabbits was used for the estimation of apoptotic clearance of pulmonary tissue. Bronchoalveolar lavage was performed in three groups of animals: experimental group-E on hypercholesterolemic diet (4% edible oil solution of crystalline cholesterol), control group-C1 on standard diet for that animal species and animals on oily diet-C2. Apoptotic detection in cytocentrifuge preparations of lung washings was evaluated by in situ TUNEL. The property of alveolar macrophages to engulf apoptotic cells was estimated by light microscopy including 300 features (related subsequent steps: adsorption, internalization and intracellular processing of free apoptotic bodies) and was evaluated by scoring and indexing method. Internalization of apoptotic bodies by alveolar macrophages, as well as free apoptotic bodies were decreased in E compared to both C1 and C2 group (p<0.01 and p<0.05 respectively). Intracellular processing of apoptotic bodies by alveolar macrophages is significantly decreased in C2 in comparison with E (p<0.05) and C1 group (p<0.01). Apoptotic capacity of pulmonary tissue is significantly decreased in C2 in comparison with C1 group (p<0.01). The results implicate that immuno-metabolic competence of pulmonary tissue might be essentially associated with tissue remodeling in pulmonary emphysema.U cilju boljeg razumevanja patogeneze plućnog emfizema, u radu je korišć en eksperimentalni model emfizema pluća na činčila kunićima za procenu apoptotskog kapaciteta plućnoga tkiva. Bronholaveolarna lavaža je urađena na tri grupe životinja: eksperimentalnoj grupi-E na hiperholesterolskoj dijeti (4% uljani rastvor kristalnog holesterola), kontrolnoj grupi-C1 na standardnoj dijeti za tu životinjsku vrstu i grupi životinja na uljanoj dijeti-C2. Određivanje apoptotskih parametara cito-centrifužnih preparata bronhoalveolarnog lavata vršeno je posle bojenja preparata TUNEL in situ citohemijskim metodom. Sposobnost alveolarnih makrofaga da odstrane apoptotske ćelije fagocitozom procenjivana je svetlosnom mikroskopijom na 300 prikaza po preparatu (prikazi uključuju: adsorpciju, internalizaciju i intracelularno procesiranje apoptotskih tela) i evaluirana metodom indeksiranja i skora. Internalizacija apoptotskih tela alveolarnim makrofazima, kao i relativni procenat slobodnih apoptotskih tela, bili su signifikantno smanjeni u E grupi poredeći sa C1 (p<0.01) i C2 grupom (p<0.05). Intracelularno procesiranje apoptotskih tela alveolarnim makrofazima bilo je signifikantno smanjeno u C2 u odnosu na E (p<0.05) i C1 grupu (p<0.01). Apoptotski kapacitet tkiva pluća bio je signifikantno smanjen u C2 u poređenju sa C1 grupom (p<0.01). Ovi rezultati ukazuju da imuno-metabolička kompetentnost plućnog tkiva može biti suštinski povezana sa remodelovanjem tkiva pluća u eksperimentalnom emfizemu pluća.nul

Collagen type I alpha 1 gene polymorphism in premature ovarian failure

Introduction. Premature ovarian failure (POF) is characterized by amenorrhea, hypergonadotropism and hypoestrogenism in women bellow 40 years. Osteoporosis is one of the late complications of POF. Objective. To correlate collagen type I alpha1 (COLIA1) gene polymorphism with bone mineral density (BMD) in women with POF. Methods. We determined the COLIA1 genotypes SS, Ss, ss in 66 women with POF. Single nucleotide polymorphism (G to T substitution) within the Sp 1-binding site in the first intron of the COLIA1 gene was assessed by polymerase chain reaction (PCR) followed by single-stranded conformation polymorphism (SSCP) analysis. Bone mineral density (BMD) was measured at the lumbar spine region by dual X-ray absorptiometry. Statistics: Kruskal-Wallis ANOVA, Chisquare test, Spearman correlation test. Results. The relative distribution of COLIA1 genotype alleles was SS - 54.4%, Ss - 41.0% and ss - 4.5%. No significant differences were found between genotype groups in body mass index, age, duration of amenorrhea or BMD. A significant positive correlation was observed between BMI and parity. Conclusion. The COLIA1 gene is just one of many genes influencing bone characteristics. It may act as a marker for differences in bone quantity and quality, bone fragility and accelerated bone loss in older women. However, in young women with POF, COLIA1 cannot identify those at higher risk for osteoporosis.Uvod. Prevremena insuficijencija jajnika (PIJ) se odlikuje amenorejom, hipergonadotropizmom i hipoestrogenijom kod žena mlađih od 40 godina. Osteoporoza je kasna komplikacija ovog stanja. Cilja rada. Cilj istraživanja je bio da se uporede genski polimorfizam kolagena tip I alfa 1 (COLIA1) sa gustinom koštane mase kod žena sa PIJ. Metode rada. Određivan je COLIA1 genotip SS, Ss i ss kod 66 žena sa PIJ pomoću eseja za reakciju lančanog umnožavanja DNK (engl. polymerase chain reaction - PCR). Polimorfizam jednog nukleotida (zamena G u T) u okviru Sp1 vezujućeg mesta u prvom intronu gena COLIA1 određivan je primenom PCR, nakon čega se pristupilo analizi konformacionog polimorfizma. Gustina koštane mase je merena na nivou lumbalnog dela kičme pomoću apsorpciometrije. Hormonske analize za folikulostimulišući hormon, luteinizirajući hormon, estradiol, prolaktin, progesteron i testosteron urađene su primenom metoda RIA. Pri statističkoj obradi podataka korišćeni su: Kraskal-Volisov (Kruskal-Wallis) ANOVA test, χ2-test i Spirmanov (Spearman) test korelacije. Rezultati. Relativna distribucija alela COLIA1 genotipa bila je: SS 54,4%, Ss 41,0% i ss 4,5%. Nije utvrđena značajna razlika između grupa prema genotipu za gustinu koštane mase, starost ispitanica, period amenoreje ili indeks telesne mase žena. Značajna pozitivna korelacija je uočena za indeks telesne mase i paritet. Zaključak. COLIA1 je samo jedan od mnogih gena koji utiču na karakteristike kosti. Kod žena starije životne dobi on može biti marker kvaliteta, kvantiteta i osetljivosti kosti. Kod mladih žena sa PIJ COLIA1 ne može da ukaže na one žene kod kojih postoji veći rizik za nastanak osteoporoze.Projekat ministarstva br. ON 17305

Changes in expression of GFAP, ApoE and APP mRNA and protein levels in the adult rat brain following cortical injury

The recovery period following cortical injury (CI) is characterized by a dynamic and highly complex interplay between beneficial and detrimental events. The aim of this study was to examine the expressions of Glial Fibrillary Acidic Protein (GFAP), Apolipoprotein E (ApoE) and Amyloid Precursor Protein (APP), all of which are involved in brain plasticity and neurodegeneration. Our results reveal that CI strongly influenced GFAP, ApoE and APP mRNA expression, as well as GFAP and ApoE protein expression. Considering the pivotal role of these proteins in the brain, the obtained results point to their potential contribution in neurodegeneration and consequent Alzheimer's disease development.Projekat ministarstva br. 17305

Supplementary material for the article: Milošev, M. Z.; Jakovljević, K.; Joksović, M. D.; Stanojković, T.; Matić, I. Z.; Perović, M.; Tešić, V.; Kanazir, S.; Mladenović, M.; Rodić, M. V.; et al. Mannich Bases of 1,2,4-Triazole3-Thione Containing Adamantane Moiety: Synthesis, Preliminary Anticancer Evaluation, and Molecular Modeling Studies. Chemical Biology and Drug Design 2017, 89 (6), 943–952. https://doi.org/10.1111/cbdd.12920

Supporting Information: [https://doi.org/10.1111/cbdd.12920]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2482]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3221

Supplementary material for the article: Milošev, M. Z.; Jakovljević, K.; Joksović, M. D.; Stanojković, T.; Matić, I. Z.; Perović, M.; Tešić, V.; Kanazir, S.; Mladenović, M.; Rodić, M. V.; et al. Mannich Bases of 1,2,4-Triazole3-Thione Containing Adamantane Moiety: Synthesis, Preliminary Anticancer Evaluation, and Molecular Modeling Studies. Chemical Biology and Drug Design 2017, 89 (6), 943–952. https://doi.org/10.1111/cbdd.12920

Supporting Information: [https://doi.org/10.1111/cbdd.12920]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2482]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3221

Collagen type I alpha 1 gene polymorphism in premature ovarian failure

Introduction. Premature ovarian failure (POF) is characterized by amenorrhea, hypergonadotropism and hypoestrogenism in women bellow 40 years. Osteoporosis is one of the late complications of POF. Objective. To correlate collagen type I alpha1 (COLIA1) gene polymorphism with bone mineral density (BMD) in women with POF. Methods. We determined the COLIA1 genotypes SS, Ss, ss in 66 women with POF. Single nucleotide polymorphism (G to T substitution) within the Sp 1-binding site in the first intron of the COLIA1 gene was assessed by polymerase chain reaction (PCR) followed by single-stranded conformation polymorphism (SSCP) analysis. Bone mineral density (BMD) was measured at the lumbar spine region by dual X-ray absorptiometry. Statistics: Kruskal-Wallis ANOVA, Chisquare test, Spearman correlation test. Results. The relative distribution of COLIA1 genotype alleles was SS - 54.4%, Ss - 41.0% and ss - 4.5%. No significant differences were found between genotype groups in body mass index, age, duration of amenorrhea or BMD. A significant positive correlation was observed between BMI and parity. Conclusion. The COLIA1 gene is just one of many genes influencing bone characteristics. It may act as a marker for differences in bone quantity and quality, bone fragility and accelerated bone loss in older women. However, in young women with POF, COLIA1 cannot identify those at higher risk for osteoporosis.Uvod. Prevremena insuficijencija jajnika (PIJ) se odlikuje amenorejom, hipergonadotropizmom i hipoestrogenijom kod žena mlađih od 40 godina. Osteoporoza je kasna komplikacija ovog stanja. Cilja rada. Cilj istraživanja je bio da se uporede genski polimorfizam kolagena tip I alfa 1 (COLIA1) sa gustinom koštane mase kod žena sa PIJ. Metode rada. Određivan je COLIA1 genotip SS, Ss i ss kod 66 žena sa PIJ pomoću eseja za reakciju lančanog umnožavanja DNK (engl. polymerase chain reaction - PCR). Polimorfizam jednog nukleotida (zamena G u T) u okviru Sp1 vezujućeg mesta u prvom intronu gena COLIA1 određivan je primenom PCR, nakon čega se pristupilo analizi konformacionog polimorfizma. Gustina koštane mase je merena na nivou lumbalnog dela kičme pomoću apsorpciometrije. Hormonske analize za folikulostimulišući hormon, luteinizirajući hormon, estradiol, prolaktin, progesteron i testosteron urađene su primenom metoda RIA. Pri statističkoj obradi podataka korišćeni su: Kraskal-Volisov (Kruskal-Wallis) ANOVA test, χ2-test i Spirmanov (Spearman) test korelacije. Rezultati. Relativna distribucija alela COLIA1 genotipa bila je: SS 54,4%, Ss 41,0% i ss 4,5%. Nije utvrđena značajna razlika između grupa prema genotipu za gustinu koštane mase, starost ispitanica, period amenoreje ili indeks telesne mase žena. Značajna pozitivna korelacija je uočena za indeks telesne mase i paritet. Zaključak. COLIA1 je samo jedan od mnogih gena koji utiču na karakteristike kosti. Kod žena starije životne dobi on može biti marker kvaliteta, kvantiteta i osetljivosti kosti. Kod mladih žena sa PIJ COLIA1 ne može da ukaže na one žene kod kojih postoji veći rizik za nastanak osteoporoze.Projekat ministarstva br. ON 17305