Wyniki wyszukiwania - Biblioteka Nauki

1

Detection of circulating breast cancer cells in peripheral blood by a two-marker reverse transcriptase-polymerase chain reaction assay.

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Fabisiewicz A. , Kulik J. , Kober P. , Brewczyńska E. , Pieńkowski T. , Siedlecki J. A.

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2004

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tom 51

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nr 3

747-755

EN

The aim of this study was to use a two-marker assay for the detection of breast cancer cells circulating in patients' blood. We have applied a PCR-based methodology to follow up the possibility of the development of metastatic disease in stage I and II patients who had undergone curative surgery. Since the number of circulating cancer cells in peripheral blood is very low, the technique for their detection needs to be not only highly sensitive, but also very specific. The reverse transcriptase-polymerase chain reaction (RT-PCR) technique may improve the sensitivity of breast cancer cell detection up to only a few cells per one million. The principle of the RT-PCR assay is to amplify a messenger RNA characteristic for breast epithelial cells in a blood sample. Since we do not expect such cells to be circulating in peripheral blood of healthy subjects, detection of the characteristic mRNA should indicate the presence of circulating breast cancer cells. We analyzed the usefulness of three mRNA markers: cytokeratin 19 (CK19), mammaglobin (hMAM) and β subunit of human chorionic gonadotropin (β-hCG) for this test. Blood samples (112) were obtained from 55 patients, in stages I and II, with or without metastasis to regional lymph nodes (N0 or N1). We found that a two-marker assay increases the sensitivity of detection of breast cancer cells in comparison with a single-marker one. Combination of two tumor-specific mRNA markers, hMAM/CK19 or β-hCG/CK19, allowed the detection of circulating breast cancer cells in 65% of N1 patients and 38% of N0 patients. By comparison, the combination hMAM/β-hCG allowed the detection of circulating breast cancer cells in the blood of 68% of N1 patients and 46% of N0 patients. Addition of the third marker did not significantly increase the detection sensitivity.

2

Application of the High Resolution Melting analysis for genetic mapping of Sequence Tagged Site markers in narrow-leafed lupin (Lupinus angustifolius L.)

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Kamel K. , Kroc M. , Święcicki W.

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tom 62

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nr 3

533-540

EN

Sequence tagged site (STS) markers are valuable tools for genetic and physical mapping that can be successfully used in comparative analyses among related species. Current challenges for molecular markers genotyping in plants include the lack of fast, sensitive and inexpensive methods suitable for sequence variant detection. In contrast, high resolution melting (HRM) is a simple and high-throughput assay, which has been widely applied in sequence polymorphism identification as well as in the studies of genetic variability and genotyping. The present study is the first attempt to use the HRM analysis to genotype STS markers in narrow-leafed lupin (Lupinus angustifolius L.). The sensitivity and utility of this method was confirmed by the sequence polymorphism detection based on melting curve profiles in the parental genotypes and progeny of the narrow-leafed lupin mapping population. Application of different approaches, including amplicon size and a simulated heterozygote analysis, has allowed for successful genetic mapping of 16 new STS markers in the narrow-leafed lupin genome.

3

Fast and easy method for total DNA extraction and gene amplification from larvae, spat and adult mussels Mytilus trossulus from the Baltic Sea

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Lasota R. , Piłczyńska J. , Williams S. T. , Wołowicz M.

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2013

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tom Vol. 42, No. 4

486--489

EN

Three nuclear DNA markers that diagnostically differentiate mussels within the Mytilus edulis complex (M. edulis, M. trossulus and M. galloprovincialis) are commonly used in taxonomic investigations: Glu5’, ITS and EFbis. As a rule, DNA extraction is performed before amplification. It is a time consuming process in the case of traditional methods based on chloroform and phenol extraction or relatively expensive using kits with ready spin columns. Moreover, DNA isolation from larvae is problematic, because of the small amount of tissue available. In this report we describe a simple, fast and inexpensive method of DNA extraction and gene amplification from larvae, spat and adults of the Baltic mussel Mytilus trossulus. The extraction method is adapted from that of Wang et al. (2006) and is based on digestion of tissue or whole animals in STE solution and direct gene amplification. On the basis of the results of routine analyses of mussels carried out in our laboratory we have concluded that the method we propose gives results that are consistent with standard methods, without requiring expensive reagents/equipment and is time saving.

4

Biotechnology of temperate fruit trees and grapevines

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Laimer M. , Mendonça D. , Maghuly F. , Marzban G. , Leopold S. , Khan M. , Balla I. , Katinger H.

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2005

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tom 52

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nr 3

673-678

EN

Challenges concerning fruit trees and grapevines as long lived woody perennial crops require adapted biotechnological approaches, if solutions are to be found within a reasonable time frame. These challenges are represented by the need for correct identification of genetic resources, with the foreseen use either in conservation or in breeding programmes. Molecular markers provide most accurate information and will be the major solution for questions about plant breeders rights. Providing healthy planting material and rapid detection of newly introduced pathogens by reliable methods involving serological and molecular biological tools will be a future challenge of increases importance, given the fact that plant material travels freely in the entire European Union. But also new breeding goals and transgenic solutions are part of the biotechnological benefits, e.g. resistance against biotic and abiotic stress factors, modified growth habits, modified nutritional properties and altered processing and storage qualities. The successful characterization of transgenic grapevines and stone fruit trees carrying genes of viral origin in different vectors constructed under ecological consideration, will be presented. Beyond technical feasibility, efficiency of resistance, environmental safety and Intellectual Property Rights, also public acceptance needs consideration and has been addressed in a specific project. The molecular determination of internal quality parameters of food can also be addressed by the use of biotechnological tools. Patient independent detection tools for apple allergens have been developed and should allow to compare fruits from different production systems, sites, and genotypes for their content of health threatening compounds.

5

β-Glucan-Mediated Alleviation of NaCl Stress in Ocimum basilicum L. in Relation to the Response of Antioxidant Enzymes and Assessment DNA Marker

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Alhasnawi A. N.

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tom Vol. 20, nr 8

90--99

EN

Salinity is one of the most important abiotic stresses which can negatively affect the plant metabolic processes in the world. This can impact the plant production, either for economic or sustenance benefits. The salinity stress can cause many physiological and biochemical changes in the plants. β-glucans are important polysaccharides, which are present in the cell walls of various cereal grains. They protect the plant responses and occur in plant suspensions. In this study, the researchers attempted to investigate various physiological mechanisms and determine the role of the β-glucans in the NaCl-mediated stress conditions on the Ocimum basilicum L. seedlings. For this purpose, they carried out an experiment for assessing various shoot and root parameters along with the antioxidant enzyme activities, proline levels and the ISSR markers. When the seedlings were exposed to the NaCl stress conditions, they showed a significant decrease in the growth parameters and an increase in the antioxidant and proline levels compared to the control seedlings grown under normal saline conditions. On the other hand, the β-glucantreated seeds, when grown under the saline stress conditions, showed better growth parameters as well as high antioxidant enzyme activities and proline levels, compared to the control and NaCl-treated plants. Furthermore, a PCR analysis was carried out using the ISSR-marker technology, which could help in evaluating the DNA fingerprints and genetic variations in the plants. The results indicated that the exogenous application of the β-glucans could protect the antioxidant enzyme activities and protect the plants against the salinity stresses, without affecting the DNA-markers without affecting the genetic variations and could be a better choice for use in DNA-markers.

6

Analysis of molecular markers as IL-12, IL-22 and IFN-γ in correlation with a clinical course in patients with psoriasis

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Kutwin M. , Migdalska-Sęk M. , Brzeziańska-Lasota E. , Zelga P. , Woźniacka A.

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nr 5

635-647

EN

ObjectivesAs a chronic, recurrent, immunologically mediated systemic disease and a common cause of dermatological problems, psoriasis is often a subject of scientific research. Skin changes located on the hands can cause difficulties and limitations in the performance of professional activities, especially manual ones. The main role in pathogenesis is played by immunological factors – improper functioning of the components of the immune system, among others, T lymphocytes and cytokines like interleukin-12 (IL-12), interleukin-22 (IL-22) and interferon gamma (IFN-γ).Material and MethodsThe obtained tissue and blood were destined for RNA isolation. The RNA was then subjected to a reverse transcription reaction. The relative gene expression level was evaluated by the real-time polymerase chain reaction for IL-12B, IL-22 and IFN-γ genes, and presented as the relative quantification (RQ) value, relative to the reference gene GAPDH. In addition, a correlation analysis of the expression level of selected genes with the clinical course of the disease, as assessed by the Psoriasis Area and Severity Index (PASI), the Body Surface Area (BSA) and the Dermatology Life Quality Index (DLQI) scores was performed.ResultsStatistical analysis confirmed a significant increase in RQ values for IL-12B, IL-22 and IFN-γ in the group of psoriatic patients vs. the control group. A positive correlation was also found between BSA and PASI and RQ for the IL-12B gene.ConclusionsIncreased expression levels of IL-12B, IL-22 and IFN-γ genes in psoriatic skin confirm that selected cytokines play an important role in the initiation and sustenance of psoriasis.

7

mRNA expressions MMP-2 and MMP-9 as potential molecular markers for distinguishing the boundary between tumour and normal tissue in basal cell carcinoma

63%

Goździalska A. , Pasek M. , Jochymek M. , Goździalska M.

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nr 1

35-53

PL

Wprowadzenie: Nieczerniakowe nowotwory skóry (NMSC) są najczęstszymi nowotworami złośliwymi, których liczba stale rośnie. BCC charakteryzuje się powolnym wzrostem, a główną cechą kliniczną tego guza jest naciekanie okolicznych tkanek i niszczenie sąsiadujących struktur. Materiał i metody: Ekspresję transkryptów mRNA dla kolagenu typu I i IV oraz MMP-2 i MMP-9 porównano w biopsjach skóry od pacjentów z BCC skóry oraz w biopsjach zdrowej skóry z marginesu guza. W badaniu wzięło udział 70 pacjentów, u których zdiagnozowano BCC. Wyniki: Stwierdzono różnice między ekspresją mRNA dla MMP-2 i MMP-9, kolagenu typu I i IV w BCC guzkowym i naciekowym w próbkach tkanek nowotworowych i tkanek prawidłowych pobranych z marginesów guza tych samych pacjentów. Wnioski: Istotnie wyższe poziomy ekspresji mRNA dla kolagenu typu I, MMP-2 i MMP-9, a także zawsze niższe poziomy ekspresji mRNA dla kolagenu typu IV w tkance nowotworowej w porównaniu z tkanką marginesu guza uzyskaną od tych samych pacjentów wykazano w obu typach BCC. Postrzegane różnice wskazują na inną rolę kolagenu I i kolagenu IV w patomechanizmie BCC.

EN

Introduction: Non-melanoma skin cancers (NMSC) are the most common malignant neoplasms, the number of which continues to increase. BCC is characterized by slow growth, its main clinical feature being that it infiltrates adjacent tissues and destroys adjacent structures. Material and methods: The expression of mRNA transcripts for collagen types I, IV and MMP-2 and MMP-9 were compared in skin biopsies from patients with BCC of the skin and in the biopsies of healthy skin from the tumour margin. The study involved seventy patients diagnosed with BCC. Results: The differences between mRNA expressions for MMP-2 and MMP-9, type I collagen and type IV collagen were investigated in nodular and infiltrative BCC both in tumour tissue samples and normal tissue samples taken from the tumour margins of the same patients. Conclusions: Significantly higher levels of mRNA expressions for type I collagen, MMP-2 and MMP-9, as well as consistently lower levels of mRNA expression for type IV collagen in tumour tissue compared to tumour margin tissue obtained from the same patients, were identified in both types of BCC. These differences indicate a different role for collagen I and collagen IV in the pathomechanism of BCC.

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Current and potential oncomarkers in diagnosing breast cancer

63%

Bilecová-Rabajdová M. , Urban P. , Grešová A. , Varga J. , Gregová K. , Stupák M. , Mareková M.

Journal of Ecology and Health

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2012

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tom R. 16, nr 3

138-143

EN

Breast cancer is a very serious disease from both an economical and social point of view, because of its high incidence and mortality, as well as its prevalence. Genetic and epigenetic changes as well as the resultant deregulation of mechanisms that affect cellular processes are reflected in the final stage, like significant variability in phenotype. These changes have multiple factors in clinical, morphological but also molecular terms, and they greatly affect the possibility of accurate diagnosis of breast cancer in its early stages. Since no combination of clinical examination, mammography and ultrasonography together with CT and MRI can achieve 100% sensitivity, clinical practice is supported with diagnostic methods based on the monitoring of tumour markers with the emphasis on molecular markers. This review describes the most common molecular markers already used for earlier clinical diagnostics and also proposes new possible markers from the tumour vascular markers group. Any new oncomarkers to improve diagnostic sensitivity and specificity will be of great benefit, and in clinical practice, they may provide an opportunity for early diagnostic and therapeutic intervention, and subsequently better prognosis for the patient as well.

PL

Rak piersi jest bardzo poważną chorobą, zarówno z punktu widzenia ekonomicznego jak i społecznego, ze względu na dużą zachorowalność i umieralność, a także na zakres jego występowania. Zmiany genetyczne i epigenetyczne, jak również wynikająca z nich deregulacja mechanizmów, które wpływają na procesy komórkowe są odzwierciedlone w ostatnim stadium choroby, jak znaczna zmienność fenotypu. Zmiany te mają wiele przyczyn z punktu widzenia klinicznego, morfologicznego, ale również molekularnego oraz znacznie wpływają na możliwość dokładnego rozpoznania raka piersi w jego wczesnych fazach. Ponieważ żadna kombinacja badań klinicznych, mammografii i USG wraz z tomografią komputerową i obrazowaniem magnetyczno-rezonansowym nie może osiągnąć 100% czułości, praktyka kliniczna jest wspierana metodami diagnostycznymi opartymi na monitoringu markerów nowotworowych, ze szczególnym uwzględnieniem markerów molekularnych. Niniejsza recenzja opisuje najbardziej typowe markery molekularne wykorzystywane we wczesnej diagnostyce klinicznej, a także proponuje nowe potencjalne markery z grupy markerów nowotworowych TVMs . Wszelkie nowe markery nowotworowe poprawiające czułość i dokładność diagnostyki będą bardzo pomocne, a w praktyce klinicznej mogą pozwolić na wczesną diagnostykę i leczenie, co polepszy także rokowania pacjenta.