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Tytuł:
Cloning and partial characterization of a gene in Bombyx mori homologous to a human adiponectin receptor
Autorzy:
Zhu, Minfeng
Chen, Keping
Wang, Yong
Guo, Zhongjian
Yin, Huijuan
Yao, Qin
Chen, Huiqin
Powiązania:
https://bibliotekanauki.pl/articles/1040735.pdf
Data publikacji:
2008
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
BmAdipoR
bioinformatics study
subcellular localization
real-time quantitative PCR
BmNPV
Opis:
In this study, we report the cloning and characteristics of an adiponectin-like receptor gene from Bombyx mori (BmAdipoR) with highly conserved deduced amino-acid sequences and similar structure to the human adiponectin receptor (AdipoR). Structural analysis of the translated cDNA suggested it encoded a membrane protein with seven transmembrane domains. BmAdipoR was found to be expressed in multiple tissues and highly expressed in Malpighian tubules, fat body and testis. BmNPV (Bombyx mori nucleopolyhedrovirus) bacmid system combined with confocal microscopy revealed that BmAdipoR was targeted to the cell membrane. We also found that infection with BmNPV did not have an effect on BmAdipoR mRNA quantity in the midgut of susceptible Bombyx mori strain (306) at 48 h, but BmAdipoR mRNA quantity increased significantly at 72 h. We concluded that BmAdipoR gene was a membrane protein ubiquitously expressed in Bombyx mori tissues and that its expression was altered by treating with BmNPV.
Źródło:
Acta Biochimica Polonica; 2008, 55, 2; 241-249
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Wewnętrzna walidacja metody kwantyfikacji dna z wykorzystaniem zestawu Quantifiler® Human DNA Quantification Kit i aparatu 7500 Real-Time PCR System wraz z oprogramowaniem Hid Real-Time PCR Analysis Software V 1.1 w Zakładzie Biologii Centralnego Laboratorium Kryminalistycznego Policji
Internal validation of a DNA quantification method using the Quantifiler® Human DNA Quantification Kit and the 7500 REAL-TIME PCR SYSTEM with the HID REAL-TIME PCR ANALYSIS SOFTWARE V 1.1 at the Biology Department of the Central Forensic Laboratory of the Police
Autorzy:
Zbieć-Piekarska, Renata
Spas, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1373985.pdf
Data publikacji:
2014
Wydawca:
Centralne Laboratorium Kryminalistyczne Policji
Tematy:
walidacja
kwantyfikacja DNA
Real-Time PCR
Quantifiler® Human DNA Quantification Kit
internal validation
quantification of DNA
Opis:
Celem przedstawionych badań było przeprowadzenie w Zakładzie Biologii Centralnego Laboratorium Kryminalistycznego Policji walidacji wewnętrznej metody kwantyfikacji DNA z wykorzystaniem zestawu Quantifiler® Human DNA Quantification Kit firmy Applied Biosystems, służącego do oznaczania całkowitej ilości DNA ludzkiego przy współpracy z aparatem 7500 Real-Time PCR System wraz z oprogramowaniem HID Real-Time PCR Analysis Software v1.1. Wyboru parametrów istotnych z punktu widzenia rutynowo prowadzonych badań dokonano na podstawie zaleceń grupy roboczej ds DNA ENFSI tj. Recommended Minimum Criteria of the Valiation of Various Aspects of the DNA Profiling Process. Ocenie poddano: czułość, liniowość, zakres metody oraz precyzję z uwzględnieniem powtarzalności i odtwarzalności wewnątrzlaboratoryjnej. Dodatkowo podjęto próbę ustalenia, czy zasadna jest kontynuacja analiz genetycznych, jeżeli wskazania aparatu 7500 Real-Time dają wynik negatywny, tj. nie wykazały obecności DNA jądrowego przy jednoczesnych prawidłowych wskazaniach dla kontroli wewnętrznej IPC. Na podstawie przeprowadzonych eksperymentów walidacyjnych stwierdzono, że zakres, w jakim walidowana metoda spełniła zadane wcześniej kryteria akceptacji, jest satysfakcjonujący, biorąc pod uwagę specyfikę badań wykonywanych w naszym laboratorium. Niemniej bardzo uważnie należy interpretować negatywne wskazania aparatu 7500 Real-Time PCR System, sugerujące brak obecności DNA jądrowego przy jednoczesnych prawidłowych wskazaniach dla kontroli wewnętrznej IPC. Okazuje się bowiem, że nie jest zasadne odstępowanie od dalszej analizy tego rodzaju próbek z uwagi na możliwość zastosowania nowych bardzo czułych zestawów do amplifikacji DNA, które niejednokrotnie pozwalają na uzyskanie pełnego profilu DNA. Podsumowując, walidacja metody kwantyfikacji DNA z wykorzystaniem zestawu Quantifiler® Human DNA Quantification Kit i aparatu 7500 Real-Time PCR System wraz z oprogramowaniem HID REAL-TIME PCR Analysis Software V1.1 została zakończona pozytywnie, w związku z czym metoda mogła zostać wdrożona i być rutynowo wykorzystywana do oznaczania całkowitej ilości DNA ludzkiego w próbkach biologicznych będących przedmiotem badań kryminalistycznych w Zakładzie Biologii CLKP.
The aim of the present study was the internal validation of a DNA quantification method employing the Quantifiler® Human DNA Quantification Kit from Applied Biosystems, used for the quantification of human total DNA, coupled with the 7500 Real-Time PCR System and the HID Real-Time PCR Analysis Software v1.1, performed at the Biology Department of the Central Forensic Laboratory of the Police. The selection of parameters relevant to the laboratory’s routine casework was made based on the ENFSI DNA Working Group recommendations included in the document Recommended Minimum Criteria for the Validation of Various Aspects of the DNA Profiling Process. The assessment regarded: sensitivity, linearity, range covered by the method and precision, including intralaboratory repeatability and reproducibility. Moreover, an attempt was made to determine whether continuation of genetic analyses is rational if the 7500 Real-Time system reads are negative, i.e. no nuclear DNA is detected, while the internal positive control (IPC) results are correct. Based on the conducted validation experiments, it was found that the extent to which the validated method met the predetermined acceptance criteria is satisfactory, taking into account the specificity of the tests conducted in our laboratory. However, any negative indications of the 7500 Real-Time PCR System, suggesting no presence of nuclear DNA along with correct results obtained for the IPC, should be interpreted very carefully. Apparently, it is not rational to discontinue further analyses of such samples due to the availability of very sensitive new DNA amplification kits which often allow obtaining a full DNA profile. To sum up, the validation of the DNA quantification method employing the Quantifiler® Human DNA Quantification Kit coupled with the 7500 Real-Time PCR System and the HID REAL-TIME PCR Analysis Software V1.1 was successful, therefore the method could be implemented and routinely used for the quantification of human total DNA in biological samples subject to forensic analyses conducted at the Biology Department of the Central Forensic Laboratory of the Police.
Źródło:
Problemy Kryminalistyki; 2014, 284
0552-2153
Pojawia się w:
Problemy Kryminalistyki
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Methods to Study the PVY Population in the Potato
Autorzy:
Yin, Zhimin
Powiązania:
https://bibliotekanauki.pl/articles/2199692.pdf
Data publikacji:
2017-06-20
Wydawca:
Instytut Hodowli i Aklimatyzacji Roślin
Tematy:
differential potato cultivars
multiplex RT-PCR
PVY strains classification
real-time RT-qPCR
sequencing
tobacco bait plants
Opis:
The PVY population in the potato has been studied continuously using tobacco bait plants in potato fields at Młochów since 1980 at two-year intervals and in potato tuber samples collected from different regions of Poland since 2001 yearly. The paper presents the combined biological, serological and molecular assays for PVY identification and strain classification. Biologically, PVY strains are defined with respect to their ability to elicit hypersensitive resistance (HR) mediated by N genes in differential potato cultivars (King Edward, Desiree and Pentland Ivory) and to symptoms in the tobacco (cultivar Samsun). Serologically, an ELISA assay based on polyclonal or monoclonal cocktail antibodies recognizes all PVY strain types, while the specif-ic monoclonal antibodies help to recognize PVYN or PVYO/PVYC strains. Multiplex RT-PCR, Real-time RT-qPCR and sequencing-based assays are used to define the PVY genome structure. In the Polish population of PVY, the strains PVYO, PVYNTN, PVYN-Wi, PVYZ-NTN and PVYE were identified, while the PVYC strain was not detected.
Źródło:
Plant Breeding and Seed Science; 2017, 75; 71-76
1429-3862
2083-599X
Pojawia się w:
Plant Breeding and Seed Science
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Changes in accumulation of heteroplasmic mitochondrial DNA and frequency of recombination via short repeats during plant lifetime in Phaseolus vulgaris
Autorzy:
Woloszynska, Magdalena
Gola, Edyta
Piechota, Janusz
Powiązania:
https://bibliotekanauki.pl/articles/1039695.pdf
Data publikacji:
2012
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Phaseolus vulgaris
sublimons
heteroplasmy
quantitative real-time PCR
plant mitochondrial genome
mtDNA recombination
Opis:
Recombination via short repeats in plant mitochondrial genomes results in sublimons - DNA molecules with a copy number much lower compared to the main mitochondrial genome. Coexistence of stoichiometrically different mitotypes, called heteroplasmy, plays an important evolutionary role, since sublimons occasionally replace the main genome resulting in a new plant phenotype. It is not clear, how frequency of recombination and sublimon production is regulated and how it is related to changes in the quantity of the main genome and sublimons. We analyzed the accumulation of two recombining main genome sequences and two resulting sublimons in apical meristems, undifferentiated tissues and leaves of different age of Phaseolus vulgaris. Copy numbers of the main genome sequences varied greatly depending on tissue type and organ age while accumulation of sublimons remained much more stable. Although the overall accumulation of plant mtDNA decreased with the leaf age, the quantity of sublimons increased relative to the main genome indicating a higher frequency of recombination via the short 314 bp repeat. Recombination was symmetrical in young developing leaves while in senescent tissues it shifted towards asymmetric events resulting in overrepresentation of one product. We propose that during plant lifetime replication and recombination frequencies change oppositely sustaining heteroplasmic compositions of the genome, which are favorable for inheritance and maintenance of complex plant mtDNA.
Źródło:
Acta Biochimica Polonica; 2012, 59, 4; 703-709
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Analizatory DNA/PCR typu lab-chip z detekcją fluorymetryczną
Lab-on-a-chip DNA/PCR analyzers with fluorometric detection
Autorzy:
Walczak, R.
Powiązania:
https://bibliotekanauki.pl/articles/154783.pdf
Data publikacji:
2010
Wydawca:
Stowarzyszenie Inżynierów i Techników Mechaników Polskich
Tematy:
real-time PCR
lab-chip
fluorescencja
fluorescence
Opis:
W artykule przedstawiono przegląd analizatorów DNA wykorzystujących technikę real-time PCR i lab-chipy. Zaprezentowano opis opracowanej metodologii i instrumentarium do detekcji fluorescencji z wykorzystaniem miniaturowych komponentów optoelektronicznych i "inteligentnego" oprogramowania analizującego. Przedstawiono dwa przykłady miniaturowych analizatorów real-time PCR/DNA opracowanych w ramach projektów europejskich i krajowych wykorzystujących lab-chipy i nowatorską metodę detekcji fluorymetrycznej.
The paper presents a novel miniaturized optical instrumentation for fluorescence excitation and detection for miniaturized real-time PCR analyzers using lab-on-a-chip (LOC) devices. Application of miniaturized semiconductor laser to fluorescence excitation, CCD-minicamera as fluorescence photodetector and specialized software for optical signal conditioning led to development of low-cost and highly sensitive optical instrumentation. To carry out full characterization of the optical instrumentation, miniaturized thermocycler co-working with LOC was built. The optical instrumentation was tested with three LOC made of different materials and technologies, giving proper detection of fluorescence signals during real-time PCR. Finally, two miniaturized devices for real-time PCR detection and identification of DNA and utilizing described here novel optical instrumentation were briefly described. The scheme and technical realization of the novel instrumentation in laboratory version is shown in Fig. 4. The detection limit of DNA was about 0,1 ng/ml (Fig. 5). It was also found that the optical instrumentation co-works with different constructions of lab-on-a-chips for DNA real-time PCR amplification (Fig. 6). The developed optical instrumentation was successfully applied to two miniaturized devices for DNA detection and identification by real-time PCR. Technical realizations of the devices and views of the lab-on-a-chips are shown in Figs. 7 and 9. In both cases, proper detection of fluorescence signals generated during real-time PCR of Campylobacter j. DNA (Fig. 8) or complementary DNA (Fig. 10) were observed. It confirmed high sensitivity of the developed optical instrumentation.
Źródło:
Pomiary Automatyka Kontrola; 2010, R. 56, nr 7, 7; 805-808
0032-4140
Pojawia się w:
Pomiary Automatyka Kontrola
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Usefulness of real-time PCR in long-term follow-up of follicular lymphoma patients
Autorzy:
Tysarowski, Andrzej
Fabisiewicz, Anna
Paszkiewicz-Kozik, Ewa
Kulik, Jadwiga
Walewski, Jan
Siedlecki, Janusz
Powiązania:
https://bibliotekanauki.pl/articles/1041125.pdf
Data publikacji:
2007
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
follicular lymphoma
minimal residual disease
molecular remission
Opis:
The aim of this study was to evaluate the usefulness of quantitative real-time PCR (RQ-PCR) for the monitoring of molecular remission in follicular lymphoma (FL) patients during long-term follow-up. RQ-PCR by the use of TaqMan® detection system is a sensitive tool to monitor minimal residual disease (MRD) in FL through amplification of the t(14;18) fusion gene during and post-therapy. In most cases the breakpoint region occurs within the major breakpoint region (MBR). Among 75 patients diagnosed with FL, cells harboring the fusion gene BCL2/JH were found in peripheral blood of 31 patients (41%). We further monitored 30 of these patients in a period varying from 6 months to 5 years by RQ-PCR. In our study the level indicating the possibility of the presence of MRD was established at more than five t(14;18)-positive cells in the background of 83000 normal cells. The results of this work also confirmed that the presence of MRD detected by RQ-PCR is an indication for careful observation of patients because of a higher risk of disease recurrence.
Źródło:
Acta Biochimica Polonica; 2007, 54, 1; 135-142
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
A simple method for extracting DNA from rhododendron plants infected with Phytophthora spp. for use in PCR
Autorzy:
Trzewik, A.
Nowak, K.J.
Orlikowska, T.
Powiązania:
https://bibliotekanauki.pl/articles/66480.pdf
Data publikacji:
2016
Wydawca:
Polska Akademia Nauk. Czytelnia Czasopism PAN
Tematy:
simple method
DNA extraction
rhododendron
leaf
plant infection
Phytophthora
polymerase chain reaction
detection
real-time PCR method
Opis:
Among the numerous protocols that describe the extraction of DNA, those relating to the isolation of DNA from infected plants, are rare. This study describes a rapid and reliable method of extracting a high quality and quantity of DNA from rhododendron leaves artificially infected with Phytophthora cactorum, P. cambivora, P. cinnamomi, P. citrophthora, and P. plurivora. The use of the modified Doyle and Doyle protocol (1987) allowed us to obtain high quantity and quality DNA (18.26 μg from 100 mg of the fresh weight of infected leaves at the ratios of A260/280 and A260/230 – 1.83 and 1.72, respectively), suitable for conventional polymerase chain reaction (PCR) and real-time PCR amplifications.
Źródło:
Journal of Plant Protection Research; 2016, 56, 1
1427-4345
Pojawia się w:
Journal of Plant Protection Research
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
HPV16 E6 polymorphism and physical state of viral genome in relation to the risk of cervical cancer in women from the south of Poland
Autorzy:
Szostek, Slawa
Zawilinska, Barbara
Klimek, Malgorzata
Kosz-Vnenchak, Magdalena
Powiązania:
https://bibliotekanauki.pl/articles/1038699.pdf
Data publikacji:
2017
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
HPV type 16 E6 variants
HPV16 physical state
real-time PCR
squamous intraepithelial lesions
invasive cervical carcinoma
Opis:
The aim of this study was to analyse the correlation between HPV16 E6 variants and the physical status of viral genome (integrated, mixed, episomal) among patients with cervical cancer (n=40) and low-grade squamous intraepithelial lesions - LSIL (n=40). The study was performed on 80 HPV16 positive samples. HPV16 E6 variants were identified using PCR and DNA sequencing. Nucleotide sequences of E6 were compared with the prototype sequence (EUR-350T). The physical state of HPV DNA was determined as the ratio of E2/E6 copy number per cell. Twelve different intratypic variants were identified as belonging to European (in 77 samples) and North-American 1 (in 3 samples) sublineages. The most prevalent non-synonymous variant was EUR-350G, which occurred with similar frequency in cervical cancer and LSIL. The frequencies of additional mutations in variants with EUR-350T or EUR-350G sequences differed significantly. For the first time, missense mutations G122A, C153T and G188A were discovered in EUR-350G variant. The integrated viral genome was predominant in women with cervical cancer. The EUR-350T prototype and EUR-350G without additional mutations variants were prevalent in cervical cancer samples with the HPV16 characterized by integrated DNA. In summary, European variants of HPV16 E6 dominated in both cancer and LSIL group. The presence of EUR-350G favoured the occurrence of additional nucleotide changes. We showed that nucleotide changes occur significantly more often in the mixed form of viral DNA and in LSIL group and that the variants without additional mutations may promote integration of HPV16 genome.
Źródło:
Acta Biochimica Polonica; 2017, 64, 1; 143-149
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods
Autorzy:
Szostek, Slawa
Biesaga, Beata
Zawilinska, Barbara
Klimek, Malgorzata
Kosz-Vnenchak, Magdalena
Powiązania:
https://bibliotekanauki.pl/articles/1038944.pdf
Data publikacji:
2015
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
human papillomavirus
squamous intraepithelial lesions
cervical carcinoma
Opis:
The aim of this study was to analyse the correlation between a new multiplex qPCR assay and a reference qPCR assay for assessment of the human papillomavirus (HPV16) load and the viral genome status. The study was performed on 100 HPV16 positive samples containing premalignant lesions and carcinomas. HPV16 E2 and E6 gene loads were assessed by two PCR methods. The load of E2 and E6 was normalized to the cell number by qPCR targeting the RNase P open reading frame. The physical state of the viral genome was determined as a ratio of E2/E6 copies number per cell. Among 100 samples analysed, there were no statistically significant differences in the E2 and E6 viral load evaluated by multiplex qPCR and qPCR, the correlation coefficients were 0.98 and 0.97, respectively. There were 19% of samples with the integrated, 73% with mixed and 8% with episomal state of viral genome detected by multiplex qPCR and 17%, 79%, 4%, respectively, found by qPCR. Prevalence of integrated and episomal forms estimated by multiplex qPCR was higher than the one obtained by qPCR (Chi2, p < 0.0001), but in samples with premalignant and malignant diagnoses no significant differences were demonstrated regardless of the methods used. Sensitivity and specificity of multiplex qPCR were 93.7% and 100% as compared with qPCR, the positive predictive value was 100%. In summary, the multiplex qPCR assay in respect of HPV16 load and the frequency of viral genome status was shown to be a sensitive and specific reference method. Simultaneous estimation of E2 and E6 genes in one reaction tube reduces the cost of testing.
Źródło:
Acta Biochimica Polonica; 2015, 62, 4; 923-928
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Optimization of the Y831C mutation detection in human DNA polymerase gamma by allelic discrimination assay
Autorzy:
Stopińska, Katarzyna
Grzybowski, Tomasz
Malyarchuk, Boris
Derenko, Miroslava
Miścicka-Śliwka, Danuta
Powiązania:
https://bibliotekanauki.pl/articles/1041222.pdf
Data publikacji:
2006
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
real-time PCR
polymerase γ
progressive external ophthalmoplegia
single nucleotide polymorphisms (SNPs)
Opis:
Many well-defined mutations in the gene for the catalytic subunit of polymerase γ (POLG1) have been found to be associated with disease, whereas the status of several mutations remains unresolved due to the conflicting reports on their frequencies in populations of healthy individuals. Here, we have developed a highly sensitive, real-time allelic discrimination assay enabling detection of the Y831C mutation in the POLG1 gene. The Y831C mutation is present in the Polish population at a frequency of 2.25%. The new assay is well suited to both extensive population studies and molecular diagnostics of POLG1.
Źródło:
Acta Biochimica Polonica; 2006, 53, 3; 591-595
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Optimization of flotation, DNA extraction and PCR methods for detection of Toxoplasma gondii oocysts in cat faeces
Autorzy:
Sroka, J.
Karamon, J.
Dutkiewicz, J.
Wójcik-Fatla, A.
Cencek, T.
Powiązania:
https://bibliotekanauki.pl/articles/2081971.pdf
Data publikacji:
2018
Wydawca:
Instytut Medycyny Wsi
Tematy:
flotation
Toxoplasma gondii
faeces
Real time PCR
nested PCR
cats
oocysts detection
Opis:
Introduction and objective. The aim of the study was to compare the effectiveness of selected oocysts concentration methods, DNA extraction protocols and PCR assays targeting the B1 gene, for the development of procedures which would be effective and useful in laboratory practice for the detection of T. gondii in faecal samples from cats. Materials and method. In order to compare the influence of the flotation fluids on microscopy and PCR detection of T. gondii, saturated solutions of saccharose, MgSO4, ZnSO4 and NaNO3 were used. To determine the sensitivity of PCR tests used: Real time PCR (RT) and nested PCR, water samples spiked with T. gondii tachyzoites and oocysts were tested. DNA was extracted using DNeasy Blood and Tissue Kit (Qiagen) (K1). The same PCR tests were used to assess the efficacy of T. gondii DNA detection in samples of cat faeces spiked with oocysts, using DNA extraction by a K1 set and a K2 set (QIamp DNA Stool Mini Kit, (Qiagen). Results. The initial results showed that NaNO3 was most useful as a flotation fluid due to the lack negative effect on the oocysts and amplification efficacy in PCR. The level of detection for water samples (100 μl) was determined as 100 tachyzoites and 1–50 oocysts in RT, and 2–20 oocysts in nested PCR. The limit of detection (LD) for stool samples (250 mg) spiked with oocysts, where the K1 set was used, determined as 250 and 5 oocysts in RT and nested PCR, respectively. For samples extracted with the K2 set, LD in RT was determined as 1–50 oocysts (depending on the variant) and 50 oocysts in nested PCR. Conclusions. The most effective methods for detection of T. gondii in cat faeces seem to be centrifugal flotation with NaNO3, followed by DNA extraction with removing of inhibitors (K2 set) and Real Time PCR targeting B1 gene.
Źródło:
Annals of Agricultural and Environmental Medicine; 2018, 25, 4; 680-685
1232-1966
Pojawia się w:
Annals of Agricultural and Environmental Medicine
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Metoda PCR w ocenie zafałszowań składu surowcowego produktów mięsnych ze strusia (Struthio camelus)
The pCR method in the assessment authenticity of meat products from ostrich (Struthio camelus)
Autorzy:
Sawicki, W.
Zych, A.
Powiązania:
https://bibliotekanauki.pl/articles/2130220.pdf
Data publikacji:
2019
Wydawca:
Polskie Towarzystwo Technologów Żywności
Tematy:
mięso strusia
identyfikacja
zafałszowania żywności
real-time PCR
Opis:
Poprawne znakowanie mięsa pozwalające konsumentowi świadomie wybrać jego rodzaj jest istotne z wielu względów – od zdrowotnych po religijne. Ponadto ze względu na różnice cen i dostępność surow- ców mięsnych pochodzących z różnych gatunków zwierząt zdarzają się przypadki fałszowania żywności. Składniki wyrobów mięsnych są wykorzystywane nieadekwatnie do informacji podanej na etykiecie. Niezgodności dotyczą zarówno użytych surowców, jak i ich zawartości w produkcie mięsnym. Mięso o wysokiej jakości zastępuje się tańszym lub jego zawartość jest mniejsza od deklarowanej. Choć obowią- zują przepisy dotyczące znakowania przetworzonych produktów mięsnych, zdarzają się przypadki wystę- powania na rynku wyrobów mięsnych zafałszowanych. W niniejszych badaniach podjęto się opracowania procedury weryfikacji autentyczności wyrobów z mięsa strusia czerwonoskórego (Struthio camelus). Zaproponowano metodę wykorzystującą technikę real-time PCR. Jak wykazano, DNA jest wystarczająco stabilne, aby wytrzymać zróżnicowaną obróbkę technologiczną. Badania prowadzono z użyciem wyrobów modelowych oraz wyrobów mięsnych (kiełbas, steków, mięsa gulaszowego) zakupionych w handlu deta- licznym. Próbki były analizowane pod względem obecności niedeklarowanych gatunków mięsa (wie- przowiny, wołowiny, mięsa z kurcząt, kaczek i gęsi) z wykorzystaniem gatunkowo specyficznych starte- rów. Na podstawie otrzymanych wyników wykazano, że opracowana procedura identyfikacji mięsa strusia może być z powodzeniem stosowana w rutynowych kontrolach, zarówno jakościowych (wykluczenie danego gatunku), jak i ilościowych (oznaczenie udziału mięsa strusia) w tego typu żywności. Wykazano ponadto, że w wyrobach przetworzonych może występować problem niedeklarowanego dodatku mięsa innego niż wskazane na etykiecie.
Źródło:
Żywność Nauka Technologia Jakość; 2019, 26, 2; 32 - 42
1425-6959
Pojawia się w:
Żywność Nauka Technologia Jakość
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Distinct expression, localization and function of two Rab7 proteins encoded by paralogous genes in a free-living model eukaryote
Autorzy:
Osińska, Magdalena
Wiejak, Jolanta
Wypych, Emilia
Bilski, Henryk
Bartosiewicz, Rafał
Wyroba, Elżbieta
Powiązania:
https://bibliotekanauki.pl/articles/1039860.pdf
Data publikacji:
2011
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
glycosylation
antipeptide antibodies
Real-Time PCR
Rab7 isotypes
RNAi
2D electrophoresis
Paramecium octaurelia
STED
Opis:
Rab7 GTPases are involved in membrane trafficking in the late endosomal/lysosomal pathway. In Paramecium octaurelia Rab7a and Rab7b are encoded by paralogous genes. Antipeptide antibodies generated against divergent C-termini recognize Rab7a of 22.5 kDa and Rab7b of 25 kDa, respectively. In 2D gel electrophoresis two immunoreactive spots were identified for Rab7b at pI about 6.34 and about 6.18 and only one spot for Rab7a of pI about 6.34 suggesting post-translational modification of Rab7b. Mass spectrometry revealed eight identical phosphorylated residues in the both proteins. ProQ Emerald staining and ConA overlay of immunoprecipitated Rab7b indicated its putative glycosylation that was further supported by a faster electrophoretic mobility of this protein upon deglycosylation. Such a post-translational modification and substitution of Ala140 in Rab7a for Ser140 in Rab7b may result in distinct targeting to the oral apparatus where Rab7b associates with the microtubular structures as revealed by STED confocal and electron microscopy. Rab7a was mapped to phagosomal compartment. Absolute qReal-Time PCR analysis revealed that expression of Rab7a was 2.6-fold higher than that of Rab7b. Upon latex internalization it was further 2-fold increased for Rab7a and only slightly for Rab7b. Post-transcriptional gene silencing of rab7a suppressed phagosome formation by 70 % and impaired their acidification. Ultrastructural analysis with double immunogold labeling revealed that this effect was due to the lack of V-ATPase recruitment to phagolysosomes. No significant phenotype changes were noticed in cells upon rab7b silencing. In conclusion, Rab7b acquired a new function, whereas Rab7a can be assigned to the phagolysosomal pathway.
Źródło:
Acta Biochimica Polonica; 2011, 58, 4; 597-607
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
The correlation analysis of WWOX expression and cancer related genes in neuroblastoma- a real time RT-PCR study
Autorzy:
Nowakowska, Magdalena
Płuciennik, Elżbieta
Wujcicka, Wioletta
Sitkiewicz, Anna
Kazanowska, Bernarda
Zielińska, Elżbieta
Bednarek, Andrzej
Powiązania:
https://bibliotekanauki.pl/articles/1039340.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
neuroblastoma
WWOX gene
LOH
real time RT-PCR
Opis:
Neuroblastoma is one of the most common paediatric cancers, described as unpredictable due to diverse patterns of behaviour. WWOX is a tumour suppressor gene whose expression is reduced in many tumour types. Loss of its expression was shown to correlate with more aggressive disease stage and mortality rate. The aim of this study was to investigate the role of the WWOX tumour suppressor gene in neuroblastoma formation. We performed real-time RT-PCR to analyse levels of WWOX expression in 22 neuroblastic tumour samples in correlation with genes involved in cell cycle regulation (CCNE1, CCND1), proliferation (MKI67), apoptosis (BCL2, BIRC5, BAX) and signal transduction (EGFR, ERBB4). We also evaluated two potential mechanisms - promoter methylation (MethylScreen method) and loss of heterozygosity (LOH) status, which could be connected with regulation of WWOX gene expression. We found a positive correlation between WWOX gene and BCL2 and HER4 JM-a and negative with cyclin D1 and E1. Our observations are consistent with previous findings and emphasise the role of WWOX in cell cycle and apoptosis regulation. Moreover, strong positive association with HER4 JM-a in this tumour type may indicate a role for WWOX in neuroblastoma cell differentiation. The presented results indicate that LOH in locus D16S3096 (located in intron 8) may be involved in the regulation of WWOX mRNAexpression. However, no association between methylation status of WWOX promoter and its expression was observed.
Źródło:
Acta Biochimica Polonica; 2014, 61, 1; 91-97
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Molekularna diagnostyka wybranych patogenów z rodzaju Phytophthora w ramach integrowanej ochrony roślin
Molecular diagnostic of Phytophthora pathogens as a tool for Integrated Pest Management
Autorzy:
Nowakowska, J.A.
Malewski, T.
Tereba, A.
Borys, M.
Oszako, T.
Powiązania:
https://bibliotekanauki.pl/articles/989551.pdf
Data publikacji:
2016
Wydawca:
Polskie Towarzystwo Leśne
Tematy:
fitopatologia
Phytophthora
wykrywanie
identyfikacja
Phytophthora alni subsp.multiformis
Phytophthora lacustris
Phytophthora taxon hungarica
metody badan
metoda real time PCR
sondy TaqMan
real time pcr
taqman
butt and fine root pathogens
phytophthora
Opis:
Traditional detection methods such as baiting or direct isolation take a long time and are incapable to handling large volume of material to be tested. The real−time PCR−based techniques are faster, more sensitive, more easily automated, and do not require post−amplification procedures. Species−specific primers for Phytophthora were designed based on the internal transcribed spacer regions (ITS) of rDNA collected from the NCBI DNA database. Primers and probes were designed using the Allele ID 7 at default search criteria. Specific probes were labeled with the reporter dyes JOE (6−carboxy−4,5−dichloro−2,7−dimethoxyfluorescein) at the 5' end and HBQ1 quencher at the 3' end (Sigma−Aldrich). The specificity of primers and fluorogenic probes was tested against genomic DNA of P. alni subsp. multiformis, P. lacustris and P. taxon hungarica. The real−time PCR reactions with the specific probes and primers yielded positive results with five concentrations of standards obtained by standard PCR reaction for corresponding Phytophthora species. The negative control (lack of DNA pathogens) yielded no amplification products. Standard curves showed a linear correlation between input DNA and cycle threshold (Ct) values with R² from 0.994 (P. alni) to 0.998 (P. taxon hungarica). The amplification efficiency of target DNA varied from 94.6% (P. alni) to 100% (P. taxon hungarica). The validation of the primers and probes designed for analysed Phytophthora species was performed on pure cultures, on soil samples from the forest nursery and declining oak stands. The designed probes displayed the high specificity of the detection of investigated species in pure cultures. The presented new molecular TaqMan probes can fully assist the integrated pest management as a powerful tool for a quick detection of above pathogenic organisms in forest nurseries. The molecular detection of harmful phytophthoras and in consequences diminishing of fungicides use for their control in forestry fully support European Union directives as well as the ‘Good plant protection practice measures' elaborated by European and Mediterranean Organisation of Plant Protection.
Źródło:
Sylwan; 2016, 160, 05; 365-370
0039-7660
Pojawia się w:
Sylwan
Dostawca treści:
Biblioteka Nauki
Artykuł
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