Amplification of Cycline D1, C-Myc and Egfr Oncogenes in Tumour Samples of Breast Cancer Patients

Background: Breast cancer is the most common form of cancer in women. It arises from multiple genetic changes in oncogenes and tumor suppressor genes. Among so far studied oncogenes relatively few, including epdermal growth factor receptor (EGFR), cyclinD1 (CCND1) and c-myc, have been found to play an important role in progression of this type of human malignancy. The aim of this study was to examine the prognostic potential of CCND1, c-myc and EGFR amplification and their possible cooperation in breast carcinogenesis. Methods: Copy number analyses of CCND1 and c-myc genes were done by TaqMan based quantitative real time PCR. Amplification status of EGFR was determined by differential PCR. Results: Amplification of CCND1, c-myc and EGFR oncogene has been found in 20.4%, 26.5% and 26.5% of breast cancer cases, respectively. Analysis showed that amplification of CCND1 oncogene was significantly associated with the stage II of disease while amplification of EGFR gene was significantly associated with overexpression of HER-2/neu. Tumour stage and expression of HER-2/neu appeared to be significant predictors of patients outcome. Stage I patients lived significantly longer then stage III patients (p=0.04) while patients with HER-2/neu overexpression had worse prognoses and lived significantly shorter (p=0.001). Finally, survival of patients who underwent hormone therapy only was significantly longer (p=0.001) then survival of the rest of patients. Conclusions: Amplification of CCND1 or EGFR oncogene is associated with the progression of breast cancer and bad prognosis. No co-ordination in amplification of CCND1, c-myc and EGFR oncogenes were established in this cohort of breast cancer patients

Stable population structure in Europe since the Iron Age, despite high mobility

Ancient DNA research in the past decade has revealed that European population structure changed dramatically in the prehistoric period (14,000–3000 years before present, YBP), reflecting the widespread introduction of Neolithic farmer and Bronze Age Steppe ancestries. However, little is known about how population structure changed from the historical period onward (3000 YBP - present). To address this, we collected whole genomes from 204 individuals from Europe and the Mediterranean, many of which are the first historical period genomes from their region (e.g. Armenia and France). We found that most regions show remarkable inter-individual heterogeneity. At least 7% of historical individuals carry ancestry uncommon in the region where they were sampled, some indicating cross-Mediterranean contacts. Despite this high level of mobility, overall population structure across western Eurasia is relatively stable through the historical period up to the present, mirroring geography. We show that, under standard population genetics models with local panmixia, the observed level of dispersal would lead to a collapse of population structure. Persistent population structure thus suggests a lower effective migration rate than indicated by the observed dispersal. We hypothesize that this phenomenon can be explained by extensive transient dispersal arising from drastically improved transportation networks and the Roman Empire’s mobilization of people for trade, labor, and military. This work highlights the utility of ancient DNA in elucidating finer scale human population dynamics in recent history

New Light of Belgrade

Abweichender Titel nach Übersetzung der Verfasserin/des VerfassersDas heute als Kulturdenkmal geschützte Kraftwerk „Energie und Licht“ in Belgrad wurde in den Jahren 1930 bis 1932 von der Schweizerischen Elektrizitäts- und Verkehrsgesellschaft Basel projektiert und errichtet. Als größtes Kraftwerk seiner Art in Belgrad trug es zu einer signifikanten Verbesserung der Versorgung der Stadt mit elektrischer Energie bei, bevor es u.a. auf Grund veralteter Technologie 1967 endgültig stillgelegt wurde. Infolge seiner architektonischen, kulturellen und historischen Werte zählt das Kraftwerk zu den bedeutenden Baudenkmälern Belgrads. Die schlichte und schnörkellose Architektur repräsentiert die Neue Sachlichkeit im Verständnis der europäischen Zwischenkriegszeit. Das Ziel der Diplomarbeit liegt in der analytischen und deskriptiven Rekonstruktion des Kraftwerks, um es im Rahmen einer denkmalgerechten Revitalisierung als attraktiven Ort im städtischen Raum zu erhalten.The “Power and Light” power station, now protected as a cultural monument in Belgrade, was designed and built between 1930 and 1932 by the Swiss Electricity and Transport Company of Basel. As the largest power station in Belgrade, it has contributed to a significant improvement in the supply of electricity to the city, before it was finally shut down in 1967, due to obsolete technology. As the result of It´s architectural, cultural and historical values, this power station is one of the most important monuments of Belgrade. The simple and unpretentious architecture represents the New Objectivity in the understanding of the European Interwar period. The goal of this master thesis is the analytical and descriptive reconstruction of the power station, within the frame of the historical monument revitalization in an order to preserve and develop it´s attractive place in the urban space of the city.14

Patogenost P. terrestris na klijancima kukuruza

Pathogenicity of P. terrestris was determined by the Knop’s medium slants method in test tubes. Isolates originated from the roots of maize (Zea mays L.), barley (Hordeum vulgare L.), Johnson grass (Sorghum halepense Pers.), sorghum (Sorghum bicolour (L.) Moench.), garlic (Allium sativum L.), onion (Allium cepa L.), barnyard millet (Echinochloa crus-galli (L.) P.Beauv.) and green foxtail (Setaria viridis (L.) P.B.). A fragment of a fungal colony, cultivated on PDA, was placed on the bottom of Knop’s medium slant in each test tube and then sterilised a maize seed was placed 2 cm away from the inoculum. After 21-day inoculation of seeds, the intensity of the development of symptoms on maize seedlings was estimated. The reddish or dark pigment on the root, mesocotyl and/or coleoptyl of seedlings was an indicator for the infection by the fungus under in vitro conditions. Based on the pathogenicity test, the isolates were classified into the following three groups: slightly (3 isolates), moderately (6 isolates) and very pathogenic (6 isolates) to maize seedlings. The obtained results show that P. terrestris, originating from different hosts, can be a maize pathogen. These results can explain the high frequency and high incidence of this fungus on maize roots in Serbia.Za utvrđivanje patogenosti izolata P. terrestris korišćena je metoda sa zakošenom Knopovom podlogom u test-epruveti. Poreklo izolata je sa korena kukuruza (Zea mays L.), ječma (Hordeum vulgare L.), divljeg sirka (Sorghum halepense Pers.), gajenog sirka (Sorghum bicolour (L.) Moench.), belog luka (Allium sativum L.), crnog luka (Allium cepa L.), korovskog prosa (Echinochloa crus-galli L.) i zelenog muhara (Setaria viridis (L.) P.B.). Fragment kolonije gljive, gajene na PDA, je stavljen na donji deo zakošene Knopove podloge u epruveti i 2 cm iznad toga sterilisano seme kukuruza. Nakon 21 dana od inokulacije semena ocenjen je intenzitet razvoja simptoma na klijanacima kukuruza. Crvenkast ili mrki pigment na korenu, mezokotilu i/ili koleoptilu klijanaca je bio indikator za infekciju gljivom u in vitro uslovima. Na osnovu testa patogenosti izolati gljive su grupisani u sledeće tri kategorije: slabo (3 izolata), srednje (6 izolata) i jako patogeni (6 izolata) za klijance kukuruza. Dobijeni rezultati ukazuju da je P. terrestris, poreklom sa različitih domaćina, patogen za kukuruz. Ovi rezultati mogu objasniti učestalost i intenzitet pojave ove gljive na korenu kukuruza u Srbiji

New Approaches With Natural Product Drugs for Overcoming Multidrug Resistance in Cancer

Resistance to chemotherapeautic drugs is one of the main obstacles to effective cancer treatment. Multidrug resistance (MDR) is defined as resistance to structurally and/or functionally unrelated drugs, and has been extensively investigated for the last three decades. There are two types of MDR: intrinsic and acquired. Tumor microenvironment selection pressure leads to the development of intrinsic MDR, while acquired resistance is a consequence of the administered chemotherapy. A central issue in chemotherapy failure is the existence of heterogeneous populations of cancer cells within one patient and patient-to-patient variability within each type of cancer. Numerous genes and pathways contribute to the development of MDR in cancer. Point mutations, gene amplification or other genetic or epigenetic changes all affect biological functions and may lead to the occurrence of MDR phenotype. Similar to the characteristics of cancerogenesis, the main features of MDR include abnormal tumor vasculature, regions of hypoxia, aerobic glycolysis, and a lower susceptibility to apoptosis. In order to achieve a lethal effect on cancer cells, drugs need to reach their intracellular target molecules. The overexpression of the efflux transporter P-glycoprotein (P-gp) in MDR cancer cells leads to decreased uptake of the drug and intracellular drug accumulation, minimising drug-target interactions. New agents being or inspired by natural products that sucessfully target these mechanisms are the main subject of this review. Two key approaches in combating MDR in cancer are discussed (i) finding agents that preserve citotoxicity toward MDR cancer cells; (ii) developing compounds that restore the cytotoxic activity of classic anticancer drugs.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}III41031]; COST Actions {[}CM1106, CM1407

Comparative analyses of individual and multiple alterations of p53, PTEN and p16 in non-small cell lung carcinoma, glioma and breast carcinoma samples

p53, p16 and PTEN are the most commonly altered tumor suppressor genes in human cancers. In the present study, we compared the presence of individual and multiple alterations of these tumor suppressors in non-small cell lung carcinoma (NSCLC), glioma and breast carcinoma, in order to evaluate specificity of each tumor type regarding the number of altered genes, as well as their combinations. We tested the mutational status, loss of heterozygosity and methylation status of these genes. Effects of gene alterations on patients survival were also assessed. In NSCLC samples, single gene alterations occurred rarely, while there was considerable increase in incidence of double gene alterations. Furthermore, coexistence of aberrant p53, PTEN and p16 was the most frequent and had significant adverse effect on the survival of NSCLC patients. On the contrary, in glioma and breast cancer specimens, substantial number of cases had aberrant single gene only. Moreover, glioma and breast carcinoma also differ in genotypes that were predominant. Specifically, in glioma samples, prevalent were co-alterations of PTEN and p16, followed by aberrant only PTEN. In breast cancer samples, alterations in all three genes as well as in p53 and p16 were the most common. Moreover, PTEN was altered exclusively with aberrant p53, with statistically significant correlation among them. Overall, our results suggest that NSCLC, glioma and breast cancer need different approaches in molecular diagnosis and treatment with particular attention toward the number and combination of targeted genes. (C) 2014 Elsevier Masson SAS. All rights reserved

Chemo-protective and regenerative effects of diarylheptanoids from the bark of black alder (Alnus glutinosa) in human normal keratinocytes

Medicinal plants are recognized from ancient times as a source of diverse therapeutic agents and many of them are used as dietary supplements. Comprehensive approaches are needed that would identify bioactive components with evident activity against specific indications and provide a better link between science (ethno-botany, chemistry, biology and pharmacology) and market Recently, the bark of black alder (Alnus glutinosa) appeared at market in the form of food supplement for treatment of different skin conditions. This study aimed to evaluate protective effects of two diarylheptanoids isolated from the bark of black alder: platyphylloside, 5(S)-1,7-di(4-hydroxyphenyl)-3-heptanone-5-O-beta-D-glucopyranoside (1) and its newly discovered analog 5(S)-1,7-di(4-hydroxyphenyl)-5-O-beta-D-[6-(E-p-coumaroylglucopyranosyl)]heptane-3-one (2) towards doxorubicin damaging activity. To that end, we employed HaCaT cells, non-cancerous human keratinocytes commonly used for skin regenerative studies. Diarylheptanoids significantly antagonized the effects of doxorubicin by lowering the sensitivity of HaCaT cells to this drug. Compound 2 prevented doxorubicin-induced cell death by activating autophagy. Both land 2 protected HaCaT cells against doxorubicin-induced DNA damage. They significantly promoted migration and affected F-actin distribution. These results indicate that chemo-protective effects of diarylheptanoids may occur at multiple subcellular levels. Therefore, diarylheptanoids 1 and 2 could be considered as protective agents for non-cancerous dividing cells during chemotherapy.Accepted version: [http://cer.ihtm.bg.ac.rs/handle/123456789/3185

Development and Validation of a Long-Term 3D Glioblastoma Cell Culture in Alginate Microfibers as a Novel Bio-Mimicking Model System for Preclinical Drug Testing

Background: Various three-dimensional (3D) glioblastoma cell culture models have a limited duration of viability. Our aim was to develop a long-term 3D glioblastoma model, which is necessary for reliable drug response studies. Methods: Human U87 glioblastoma cells were cultured in alginate microfibers for 28 days. Cell growth, viability, morphology, and aggregation in 3D culture were monitored by fluorescent and confocal microscopy upon calcein-AM/propidium iodide (CAM/PI) staining every seven days. The glioblastoma 3D model was validated using temozolomide (TMZ) treatments 3 days in a row with a recovery period. Cell viability by MTT and resistance-related gene expression (MGMT and ABCB1) by qPCR were assessed after 28 days. The same TMZ treatment schedule was applied in 2D U87 cell culture for comparison purposes. Results: Within a long-term 3D model system in alginate fibers, U87 cells remained viable for up to 28 days. On day 7, cells formed visible aggregates oriented to the microfiber periphery. TMZ treatment reduced cell growth but increased drug resistance-related gene expression. The latter effect was more pronounced in 3D compared to 2D cell culture. Conclusion: Herein, we described a long-term glioblastoma 3D model system that could be particularly helpful for drug testing and treatment optimization

Prolonged survival after neoadjuvant chemotherapy related with specific molecular alterations in the patients with nonsmall-cell lung carcinoma

Lung cancer is the most common cause of neoplasia-related death worldwide. Accounting for approximately 80\% of all lung carcinomas, the non-small cell lung carcinoma (NSCLC) is the most common clinical form with its two predominant histological types, adenocarcinoma (ADC) and squamous cell carcinoma (SCC). Although surgical resection is the most favorable treatment for patients with NSCLC, relapse is still high, so neoadjuvant chemotherapy (NAC) is an accepted treatment modality. In this study we examined whether some of the key molecules associated with the RAS/RAF/MEK/ERK and PI3K/AKT/mTOR signaling pathways could have predictive and prognostic value for the NAC application. To that end we examined the expression status of PTEN, pAKT, pERK and loss of heterozygosity (LOH) of PTEN in two groups of NSCLC patients, those who received and those who did not receive NAG LOH PTEN and low pERK expression is shown to be correlated with the longest survival of patients with SCC and ADC, respectively, who received NAC. These results point that the application of NAC is beneficial in the NSCLC patients with specific molecular alterations which could further help to improve constant search for the druggable molecular targets used in personalized therapy. (C) 2014 Elsevier Inc. All rights reserved.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}III41031