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1.
SINGLE NUCLEOTIDE POLYMORPHISMS OF THE CHROMOSOME SEGREGATION GENES INVOLVED IN THE DEVELOPMENT OF GASTRIC CANCER
Marija Rogar, 2016, doktorska disertacija

Opis: INTRODUCTION. Gastric cancer represents the fifth most frequently diagnosed cancer in the world. Despite numerous research studies, mechanisms leading to disease are poorly known and unclear. At the molecular level, many changes are involved in the development of gastric cancer, including malfunction of chromosome segregation genes. These abnormalities can lead to chromosomal instability (CIN). Segregation gene function can be affected by the low penetrance errors which include polymorphisms. AIM. The aim of this study is to examine the effect of selected polymorphisms in specific segregation genes on gastric cancer development. HYPOTHESIS. The study focused on exploring genotypes of selected polymorphisms in specific mitotic segregation genes. Those that differ significantly between the subjects and the healthy control population, may be associated with higher risk for developing gastric cancer or with certain clinical and histopathological characteristics, and may have effect on the survival of gastric cancer patients. METHODS. 30 polymorphisms in genes BUB1B, CASC5, ESPL1, PTTG1, SMC1A, TPX2, TTK and ZWINT were included in the study. Subjects were compared with the control group. Polymorphisms were determined using quantitative real-time PCR (qPCR), restriction fragment length polymorphism (RFLP) and DNA sequencing. RESULTS. The association between polymorphisms rs2277559 (BUB1B), rs2241666 (ZWINT), rs11858113 (CASC5) and rs11855334 (CASC5) and increased risk of developing gastric cancer in male population was determined. As concerning rs11855334, statistically significant difference was also observed in the genotype distribution between the whole population of subjects and controls. The association between the genotypes of polymorphisms (in gene BUB1B) rs2277559, rs2290551, rs1801376, rs1047130, rs1565866, rs2277560 and Lauren classification was recognized. Genotypes of polymorphisms rs1801376 (BUB1B), rs11855334 (CASC5), rs2241666 (ZWINT), rs2910101 (PTTG1) and rs1047130 (BUB1B) are linked to different tumour differentiation grades. Survival analysis revealed association between the lymph node involvement and perineural invasion. Statistically higher frequencies of haplotypes G-A-G-T-G-G-A, G-G-A-G-A-A-G and A-G-G-T-A-G-A in gene BUB1B and of haplotypes A-A-A-C and C-C-G-T in gene ESPL1 were observed in gastric cancer patients, whereas haplotypes A-C-A-T and C-A-G-T in gene ESPL1 were significantly more frequent in the control group. Association with gastric cancer was not noted with other polymorphisms. CONCLUSIONS. The association between specific polymorphisms of selected chromosome segregation genes and gastric cancer was recognized. Findings could provide guidelines for further research and polymorphisms linked to gastric cancer could serve as potential diagnostic and prognostic biomarkers.
Najdeno v: ključnih besedah
Ključne besede: gastric cancer, chromosomal instability, polymorphisms, segregation genes
Objavljeno: 27.07.2016; Ogledov: 2985; Prenosov: 228
.pdf Polno besedilo (2,87 MB)

2.
Development and Validation of a High- Performance Liquid Chromatography– Tandem Mass Spectrometry Method for the Simultaneous Determination of Irinotecan and Its Main Metabolites in Human Plasma and Its Application in a Clinical Pharmacokinetic Study
Giuseppe Toffoli, Elena Marangon, Elisa Mazzega, Bianca Posocco, 2015, izvirni znanstveni članek

Opis: Irinotecan is currently used in several cancer regimens mainly in colorectal cancer (CRC). This drug has a narrow therapeutic range and treatment can lead to side effects, mainly neutropenia and diarrhea, frequently requiring discontinuing or lowering the drug dose. A wide inter-individual variability in irinotecan pharmacokinetic parameters and pharmacodynamics has been reported and associated to patients’ genetic background. In particular, a polymorphism in the UGT1A1 gene (UGT1A1*28) has been linked to an impaired detoxification of SN-38 (irinotecan active metabolite) to SN-38 glucuronide (SN-38G) leading to increased toxicities. Therefore, therapeutic drug monitoring of irinotecan, SN-38 and SN-38G is recommended to personalize therapy. In order to quantify simultaneously irinotecan and its main metabolites in patients’ plasma, we developed and validated a new, sensitive and specific HPLC–MS/MS method applicable to all irinotecan dosages used in clinic. This method required a small plasma volume, addition of camptothecin as internal standard and simple protein precipitation. Chromatographic separation was done on a Gemini C18 column (3 μM, 100 mm x 2.0 mm) using 0.1% acetic acid/bidistilled water and 0.1% acetic acid/acetonitrile as mobile phases. The mass spectrometer worked with electrospray ionization in positive ion mode and selected reaction monitoring. The standard curves were linear (R20.9962) over the concentration ranges (10–10000 ng/mL for irinotecan, 1–500 ng/mL for SN-38 and SN-38G and 1–5000 ng/mL for APC) and had good back-calculated accuracy and precision. The intra- and inter-day precision and accuracy, determined on three quality control levels for all the analytes, were always <12.3% and between 89.4% and 113.0%, respectively. Moreover, we evaluated this bioanalytical method by re-analysis of incurred samples as an additional measure of assay reproducibility. This method wassuccessfully applied to a pharmacokinetic study in metastatic CRC patients enrolled in a genotype-guided phase Ib study of irinotecan administered in combination with 5-fluorouracil/ leucovorin and bevacizumab.
Najdeno v: ključnih besedah
Povzetek najdenega: ...Irinotecan is currently used in several cancer regimens mainly in colorectal cancer (CRC). This...
Ključne besede: Pharmacokinetic, pharmacodynamics, phase I clinical study, colorectal cancer, Mass spectrometry, pharmacogenetics
Objavljeno: 21.03.2017; Ogledov: 1715; Prenosov: 0
.pdf Polno besedilo (1018,06 KB)

3.
Nanobody technology: principles & applications
Ario De Marco, predavanje na tuji univerzi

Opis: Antibodies possess unattainable capacity to bind selectively and at high affinity their cognates. For this reason they have been largely used in applications which rely on specific molecular recognition. Biosensors, nanoparticles, and even cells can be functionalized with antibodies for improving sensitivity and target specificity. However, conventional antibodies (IgGs) are large molecules (150 kDa) that are difficult to engineer. In the last years, antibody fragments have become more and more popular as an effective alternative and specifically nanobodies raised enthusiasm because of their minimal mass (14 kDa), high stability, relative similarity to human sequences, and simplified mutagenesis. Pre-immune nanobody libraries have the further advantage of enabling blind selection for antigens that can be used to discriminate between subpopulations of cells and vesicles. The panning can be performed directly on intact cells and the resulting binders are specific for the native antigen conformation
Najdeno v: ključnih besedah
Povzetek najdenega: ...Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering...
Ključne besede: Nanobodies, in vitro panning, cancer biomarkers, recombinant antibody engineering
Objavljeno: 03.05.2017; Ogledov: 2225; Prenosov: 0
.pdf Polno besedilo (37,92 KB)

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Urinary Extracellular Vesicle Biomarkers in Urological Cancers: From Discovery Towards Clinical Implementation
Bert Dhondt, Jan Van Deun, Silke Vermaerke, Ario de Marco, Nicolaas Lumen, Olivier De Wever, An Hendrix, 2018, pregledni znanstveni članek

Najdeno v: ključnih besedah
Povzetek najdenega: ...Extracellular vesicles, cancer biomarkers, diagnostics...
Ključne besede: Extracellular vesicles, cancer biomarkers, diagnostics
Objavljeno: 04.05.2018; Ogledov: 1356; Prenosov: 0
.pdf Polno besedilo (809,91 KB)

6.
Connective tissue and diseases: from morphology to proteomics towards the development of new therapeutic appproach
Daniela Quaglino, Federica Boraldi, Giulia Annovi, Deanna Guerra, Ivonne Pasquali Ronchetti, 2009, pregledni znanstveni članek

Opis: Connective tissue consists of cells separated by the extracellular matrix, whose composition and amount vary according to age, to functional requirements, and to the presence of pathologic conditions. Within this non-random macromolecular assembly, collagens, elastin, proteoglycans and structural glycoproteins are mutually interdependent and modifications of one component, by extrinsic (environmental) and/or intrinsic (systemic, genetic, age-related) factors, may have consequences on the tissue as a whole. Since decades, different microscopical techniques have been applied mainly for diagnostic purposes and for detailed descriptions of changes occurring in cells and in matrix components. More recently, in order to dissect the molecular complexity of the matrix network, to analyse the interactions between cells and matrix and to look for modulators of cell phenotype, histomorphologic investigations have been implemented with proteomic studies that allow to identify possible diagnostic markers, and to better understand patho-mechanisms enabling the design of novel therapeutic strategies. Therefore, the progressively expanding, although incomplete, knowledge on connective tissue biology, sheds new light on the pathogenesis of diseases affecting single molecules (i.e. collagenopathies, mucopolysaccharidoses, elastinopathies) and discloses the importance of matrix components as fundamental regulators of cell phenotype, in relation, for instance, to the aging process and/or to cancer development and progression. Few examples will be presented demonstrating the promises of proteomics as a technique leading to the discovery of new therapies and possibly to the development of individualized treatments for a better patient care.
Najdeno v: ključnih besedah
Povzetek najdenega: ...for instance, to the aging process and/or to cancer development and progression. Few examples will be...
Ključne besede: pathology, proteomics, fibrosis, rheumatology, cancer
Objavljeno: 23.08.2019; Ogledov: 486; Prenosov: 0
.pdf Polno besedilo (967,65 KB)

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DEVELOPMENT OF A NOVEL PCR-BASED ASSAY FOR HIGH-RISK HUMAN PAPILLOMAVIRUS DETECTION AND GENOTYPING IN SELF COLLECTED CERVICOVAGINAL SAMPLES: A NEW POSSIBILITY FOR THE CERVICAL CANCER SCREENING
Alice Avian, 2020, doktorska disertacija

Opis: Human Papillomavirus (HPV) infection is the causative agent for the invasive cervical cancer and its precancerous lesions, furthermore, there are growing evidences of HPV being a relevant factor in other anogenital cancers as well as head and neck cancers. Most sexually active women become infected with HPV at least once in their lifetime, but less than 10% of women becomes persistently infected, and it is precisely the persistent infection that contributes to the development of cervical cancer. The preventive effect of cervical cancer screening largely depends in the high women participation and coverage; indeed, a large number of cervical cancers diagnoses normally arise among under-screened and unscreened women. Increase in the screening coverage is essential to improve the effectiveness of cervical screening programmes. The main purpose of this PhD project was to solve some of the most relevant problems in the cervical cancer screening programmes, as the increase of cost-effectiveness and the amelioration of the screening coverage. My work was focused on the development and validation of the first Ulisse BioMed S.p.A. product, the HPV Selfy™ test, an innovative PCR-based kit for the direct detection and genotyping of 12 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59) and 2 possible/probable high-risk (66 and 68), specifically optimized for the analysis of self-collected vaginal specimens. The core of this innovative test is based on high-resolution melting (HRM) analysis, a recently developed technique for fast, high-throughput post-PCR analysis of variance in nucleic acid sequences, that characterizes the amplicons by studying thermal denaturation of double-stranded DNA. Based on this approach and through the design of different HPV type-specific primer pairs and the development of a specific master mix, unique melting peaks in a single fluorescence channel were obtained, allowing the multiple detection and genotyping of 14 HPV types in a single PCR well. Three different clinical studies have been carried out to validate the assay on the vaginal self-collected samples with truly amazing results regarding the assay’s performance, but also for self-sampling acceptability by women. Moreover, data collected in these studies suggest a future possible use of this test for the hard-to-reach women, as an alternative of the conventional clinician-collected sample, in order to increase the cervical cancer screening coverage.
Najdeno v: ključnih besedah
Ključne besede: Human Papillomavirus, HPV test, cervical cancer screening, prevention, diagnostic test, High resolution melting, HRM, genotyping, PCR, Self-sampling, clinical validation.
Objavljeno: 17.06.2020; Ogledov: 299; Prenosov: 6
.pdf Polno besedilo (37,88 MB)

9.
Covalent Calix[4]arene Nanocapsules for Efficient Camptothecin Delivery
Ali Trabolsi, Tina Skorjanc, Dinesh Shetty, 2019, objavljeni povzetek znanstvenega prispevka na konferenci

Najdeno v: ključnih besedah
Ključne besede: drug delivery, calixarene, nanocapsule, self-assembly, camptothecin, breast cancer
Objavljeno: 10.09.2020; Ogledov: 117; Prenosov: 0
.pdf Polno besedilo (1,99 MB)

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