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1.
Plant Cell Environ ; 47(6): 2228-2239, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38483021

RESUMEN

The selection of oviposition sites by female moths is crucial in shaping their progeny performance and survival, and consequently in determining insect fitness. Selecting suitable plants that promote the performance of the progeny is referred to as the Preference-Performance hypothesis (or 'mother-knows-best'). While root infestation generally reduces the performance of leaf herbivores, little is known about its impact on female oviposition. We investigated whether maize root infestation by the Western corn rootworm (WCR) affects the oviposition preference and larval performance of the European corn borer (ECB). ECB females used leaf volatiles to select healthy plants over WCR-infested plants. Undecane, a compound absent from the volatile bouquet of healthy plants, was the sole compound to be upregulated upon root infestation and acted as a repellent for first oviposition. ECB larvae yet performed better on plants infested below-ground than on healthy plants, suggesting an example of 'bad motherhood'. The increased ECB performance on WCR-infested plants was mirrored by an increased leaf consumption, and no changes in the plant primary or secondary metabolism were detected. Understanding plant-mediated interactions between above- and below-ground herbivores may help to predict oviposition decisions, and ultimately, to manage pest outbreaks in the field.


Asunto(s)
Larva , Mariposas Nocturnas , Oviposición , Hojas de la Planta , Raíces de Plantas , Compuestos Orgánicos Volátiles , Zea mays , Animales , Oviposición/efectos de los fármacos , Zea mays/fisiología , Zea mays/parasitología , Compuestos Orgánicos Volátiles/metabolismo , Compuestos Orgánicos Volátiles/farmacología , Mariposas Nocturnas/fisiología , Femenino , Larva/fisiología , Raíces de Plantas/parasitología , Raíces de Plantas/fisiología , Hojas de la Planta/fisiología , Herbivoria
2.
Avian Pathol ; 53(4): 257-263, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38353105

RESUMEN

The aim of the present study was to monitor the dynamics and to measure the safety and efficacy of a live, attenuated, thermosensitive Mycoplasma anserisalpingitidis vaccine candidate, namely MA271, in geese breeder flocks under field conditions. Two rearing flocks were vaccinated with MA271 at 4 weeks of age and boosted at 24 weeks of age by cloaca inoculation (1 ml) and eye-dropping (60 µl). The geese then were transported to multi-aged breeding farms. Two breeding flocks served as controls. Colonization of the cloaca by MA271 showed 75% maximum prevalence between 4 and 6 weeks after the first vaccination. Then the prevalence decreased to 25% until the cooler, humid fall months which coincided with the booster vaccination. Boosting raised cloacal colonization to 100%. No clinical signs were observed in the vaccinated birds. After transportation to five multi-aged breeding farms, the wild-type strain appeared as well as MA271 in three flocks. In one flock, the wild-type strain completely displaced MA271, while in one flock only MA271 was detected. Only wild-type strains were detected in the control flocks; however, due to an HPAI outbreak, both flocks were exterminated before the end of the study. Based on the available data, the median percentage of infertile eggs was 3.7-5.1% in the MA271 vaccinated flocks, and 7.7% in the non-vaccinated flock. In conclusion, MA271 can colonize the cloaca of geese under field conditions. MA271 proved to be safe and presumably protects against M. anserisalpingitidis-induced reproduction losses.


Asunto(s)
Vacunas Bacterianas , Gansos , Infecciones por Mycoplasma , Enfermedades de las Aves de Corral , Vacunas Atenuadas , Animales , Enfermedades de las Aves de Corral/prevención & control , Enfermedades de las Aves de Corral/microbiología , Infecciones por Mycoplasma/veterinaria , Infecciones por Mycoplasma/prevención & control , Vacunas Atenuadas/inmunología , Vacunas Atenuadas/administración & dosificación , Vacunas Bacterianas/inmunología , Vacunación/veterinaria , Cloaca/microbiología , Mycoplasma/inmunología , Femenino , Granjas
3.
Cytometry A ; 103(3): 198-207, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-35880846

RESUMEN

The emergence and fast advance of digital pathology allows the acquisition, digital storage, interactive recall and analysis of morphology at the tissue level. When applying immunohistochemistry, it also affords the correlation of morphology with the expression of one or two specific molecule of interest. The rise of fluorescence pathology scanners expands the number of detected molecules based on multiplex labeling. The Pannoramic Confocal (created by 3DHistech, Hungary) is a first-of-the-kind digital pathology scanner that affords not only multiplexed fluorescent detection on top of conventional transmission imaging, but also confocality. We have benchmarked this scanner in terms of stability, precision, light efficiency, linearity and sensitivity. X-Y stability and relocalisation precision were well below resolution limit (≤50 nm). Light throughput in confocal mode was 4-5 times higher than that of a point scanning confocal microscope, yielding similar calculated confocal intensities but with the potential for improving signal to noise ratio or scan speed. Response was linear with R2 ≥ 0.9996. Calibrated measurements showed that using indirect labeling ≥2000 molecules per cell could be well detected and imaged on the cell surface. Both standard-based and statistical post-acquisition flatfield corrections are implemented. We have also measured the point spread function (PSF) of the instrument. The dimensions of the PSF are somewhat larger and less symmetric than of the theoretical PSF of a conventional CLSM, however, the spatial homogeneity of these parameters allows for obtaining a specific system PSF for each optical path and using it for optional on-the-fly deconvolution. In conclusion, the Pannoramic Confocal provides sensitive, quantitative widefield and confocal detection of multiplexed fluorescence signals, with optical sectioning and 3D reconstruction, in addition to brightfield transmission imaging. High speed scanning of large samples, analysis of tissue heterogeneity, and detection of rare events open up new ways for quantitatively analyzing tissue sections, organoid cultures or large numbers of adherent cells.


Asunto(s)
Microscopía , Patología Molecular , Microscopía/métodos , Colorantes
4.
Sensors (Basel) ; 23(22)2023 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-38005629

RESUMEN

As the field of routine pathology transitions into the digital realm, there is a surging demand for the full automation of microscope scanners, aiming to expedite the process of digitizing tissue samples, and consequently, enhancing the efficiency of case diagnoses. The key to achieving seamless automatic imaging lies in the precise detection and segmentation of tissue sample regions on the glass slides. State-of-the-art approaches for this task lean heavily on deep learning techniques, particularly U-Net convolutional neural networks. However, since samples can be highly diverse and prepared in various ways, it is almost impossible to be fully prepared for and cover every scenario with training data. We propose a data augmentation step that allows artificially modifying the training data by extending some artifact features of the available data to the rest of the dataset. This procedure can be used to generate images that can be considered synthetic. These artifacts could include felt pen markings, speckles of dirt, residual bubbles in covering glue, or stains. The proposed approach achieved a 1-6% improvement for these samples according to the F1 Score metric.


Asunto(s)
Artefactos , Procesamiento de Imagen Asistido por Computador , Procesamiento de Imagen Asistido por Computador/métodos , Redes Neurales de la Computación , Automatización
5.
BMC Cancer ; 22(1): 605, 2022 Jun 02.
Artículo en Inglés | MEDLINE | ID: mdl-35655145

RESUMEN

BACKGROUND: Hypomethylation of long interspersed nuclear element 1 (LINE-1) is characteristic of various cancer types, including colorectal cancer (CRC). Malfunction of several factors or alteration of methyl-donor molecules' (folic acid and S-adenosylmethionine) availability can contribute to DNA methylation changes. Detection of epigenetic alterations in liquid biopsies can assist in the early recognition of CRC. Following the investigations of a Hungarian colon tissue sample set, our goal was to examine the LINE-1 methylation of blood samples along the colorectal adenoma-carcinoma sequence and in inflammatory bowel disease. Moreover, we aimed to explore the possible underlying mechanisms of global DNA hypomethylation formation on a multi-level aspect. METHODS: LINE-1 methylation of colon tissue (n = 183) and plasma (n = 48) samples of healthy controls and patients with colorectal tumours were examined with bisulfite pyrosequencing. To investigate mRNA expression, microarray analysis results were reanalysed in silico (n = 60). Immunohistochemistry staining was used to validate DNA methyltransferases (DNMTs) and folate receptor beta (FOLR2) expression along with the determination of methyl-donor molecules' in situ level (n = 40). RESULTS: Significantly decreased LINE-1 methylation level was observed in line with cancer progression both in tissue (adenoma: 72.7 ± 4.8%, and CRC: 69.7 ± 7.6% vs. normal: 77.5 ± 1.7%, p ≤ 0.01) and liquid biopsies (adenoma: 80.0 ± 1.7%, and CRC: 79.8 ± 1.3% vs. normal: 82.0 ± 2.0%, p ≤ 0.01). However, no significant changes were recognized in inflammatory bowel disease cases. According to in silico analysis of microarray data, altered mRNA levels of several DNA methylation-related enzymes were detected in tumours vs. healthy biopsies, namely one-carbon metabolism-related genes-which met our analysing criteria-showed upregulation, while FOLR2 was downregulated. Using immunohistochemistry, DNMTs, and FOLR2 expression were confirmed. Moreover, significantly diminished folic acid and S-adenosylmethionine levels were observed in parallel with decreasing 5-methylcytosine staining in tumours compared to normal adjacent to tumour tissues (p ≤ 0.05). CONCLUSION: Our results suggest that LINE-1 hypomethylation may have a distinguishing value in precancerous stages compared to healthy samples in liquid biopsies. Furthermore, the reduction of global DNA methylation level could be linked to reduced methyl-donor availability with the contribution of decreased FOLR2 expression.


Asunto(s)
Adenoma , Neoplasias Colorrectales , Receptor 2 de Folato , Enfermedades Inflamatorias del Intestino , Adenoma/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , ADN/metabolismo , Metilación de ADN , Receptor 2 de Folato/genética , Receptor 2 de Folato/metabolismo , Ácido Fólico , Humanos , Biopsia Líquida , ARN Mensajero/metabolismo , S-Adenosilmetionina/metabolismo
6.
PLoS Comput Biol ; 17(9): e1009327, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34534207

RESUMEN

DNA methylation provides one of the most widely studied biomarkers of ageing. Since the methylation of CpG dinucleotides function as switches in cellular mechanisms, it is plausible to assume that by proper adjustment of these switches age may be tuned. Though, adjusting hundreds of CpG methylation levels coherently may never be feasible and changing just a few positions may lead to biologically unstable state. A prominent example of methylation-based age estimators is provided by Horvath's clock, based on 353 CpG dinucleotides, showing a high correlation (not necessarily causation) with chronological age across multiple tissue types. On this small subset of CpG dinucleotides we demonstrate how the adjustment of one methylation level leads to a cascade of changes at other sites. Among the studied subset, we locate the most important CpGs (and related genes) that may have a large influence on the rest of the sub-system. According to our analysis, the structure of this network is way more hierarchical compared to what one would expect based on ensembles of uncorrelated connections. Therefore, only a handful of CpGs is enough to modify the system towards a desired state. When propagation of the change over the network is taken into account, the resulting modification in the predicted age can be significantly larger compared to the effect of isolated CpG perturbations. By adjusting the most influential single CpG site and following the propagation of methylation level changes we can reach up to 5.74 years in virtual age reduction, significantly larger than without taking into account of the network control. Extending our approach to the whole methylation network may identify key nodes that have controller role in the ageing process.


Asunto(s)
Envejecimiento/genética , Metilación de ADN , Islas de CpG , Humanos
7.
Sensors (Basel) ; 22(18)2022 Sep 14.
Artículo en Inglés | MEDLINE | ID: mdl-36146303

RESUMEN

Ploidy analysis is the fundamental method of measuring DNA content. For decades, the principal way of conducting ploidy analysis was through flow cytometry. A flow cytometer is a specialized tool for analyzing cells in a solution. This is convenient in laboratory environments, but prohibits measurement reproducibility and the complete detachment of sample preparation from data acquisition and analysis, which seems to have become paramount with the constant decrease in the number of pathologists per capita all over the globe. As more open computer-aided systems emerge in medicine, the demand for overcoming these shortcomings, and opening access to even more (and more flexible) options, has also emerged. Image-based analysis systems can provide an alternative to these types of workloads, placing the abovementioned problems in a different light. Flow cytometry data can be used as a reference for calibrating an image-based system. This article aims to show an approach to constructing an image-based solution for ploidy analysis, take measurements for a basic comparison of the data produced by the two methods, and produce a workflow with the ultimate goal of calibrating the image-based system.


Asunto(s)
ADN de Neoplasias , Ploidias , Calibración , ADN de Neoplasias/genética , Citometría de Flujo , Procesamiento de Imagen Asistido por Computador , Reproducibilidad de los Resultados
8.
Int J Mol Sci ; 23(19)2022 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-36232908

RESUMEN

Long interspersed nuclear element 1 (LINE-1) bisulfite pyrosequencing is a widely used technique for genome-wide methylation analyses. We aimed to investigate the effects of experimental and biological factors on its results to improve the comparability. LINE-1 bisulfite pyrosequencing was performed on colorectal tissue (n = 222), buffy coat (n = 39), and plasma samples (n = 9) of healthy individuals and patients with colorectal tumors. Significantly altered methylation was observed between investigated LINE-1 CpG positions of non-tumorous tissues (p ≤ 0.01). Formalin-fixed, paraffin-embedded biopsies (73.0 ± 5.3%) resulted in lower methylation than fresh frozen samples (76.1 ± 2.8%) (p ≤ 0.01). DNA specimens after long-term storage showed higher methylation levels (+3.2%, p ≤ 0.01). In blood collection tubes with preservatives, cfDNA and buffy coat methylation significantly changed compared to K3EDTA tubes (p ≤ 0.05). Lower methylation was detected in older (>40 years, 76.8 ± 1.7%) vs. younger (78.1 ± 1.0%) female patients (p ≤ 0.05), and also in adenomatous tissues with MTHFR 677CT, or 1298AC mutations vs. wild-type (p ≤ 0.05) comparisons. Based on our findings, it is highly recommended to consider the application of standard DNA samples in the case of a possible clinical screening approach, as well as in experimental research studies.


Asunto(s)
Ácidos Nucleicos Libres de Células , Neoplasias Colorrectales , Anciano , Factores Biológicos , Biopsia , Ácidos Nucleicos Libres de Células/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , ADN/genética , Metilación de ADN , Femenino , Formaldehído , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Biopsia Líquida , Elementos de Nucleótido Esparcido Largo/genética , Masculino , Sulfitos
9.
Int J Mol Sci ; 23(7)2022 Mar 29.
Artículo en Inglés | MEDLINE | ID: mdl-35409133

RESUMEN

Monitoring the therapeutic response of colorectal cancer (CRC) patients is crucial to determine treatment strategies; therefore, we constructed a liquid biopsy-based approach for tracking tumor dynamics in non-metastatic (nmCRC) and metastatic (mCRC) patients (n = 55). Serial blood collections were performed during chemotherapy for measuring the amount and the global methylation pattern of cell-free DNA (cfDNA), the promoter methylation of SFRP2 and SDC2 genes, and the plasma homocysteine level. The average cfDNA amount was higher (p < 0.05) in nmCRC patients with recurrent cancer (30.4 ± 17.6 ng) and mCRC patients with progressive disease (PD) (44.3 ± 34.5 ng) compared to individuals with remission (13.2 ± 10.0 ng) or stable disease (12.5 ± 3.4 ng). More than 10% elevation of cfDNA from first to last sample collection was detected in all recurrent cases and 92% of PD patients, while a decrease was observed in most patients with remission. Global methylation level changes indicated a decline (75.5 ± 3.4% vs. 68.2 ± 8.4%), while the promoter methylation of SFRP2 and SDC2 and homocysteine level (10.9 ± 3.4 µmol/L vs. 13.7 ± 4.3 µmol/L) presented an increase in PD patients. In contrast, we found exact opposite changes in remission cases. Our study offers a more precise blood-based approach to monitor the treatment response to different chemotherapies than the currently used markers.


Asunto(s)
Ácidos Nucleicos Libres de Células , Neoplasias Colorrectales , Biomarcadores de Tumor/genética , Ácidos Nucleicos Libres de Células/genética , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/cirugía , Metilación de ADN , Homocisteína , Humanos , Biopsia Líquida , Recurrencia Local de Neoplasia/genética
10.
Sensors (Basel) ; 21(21)2021 Oct 26.
Artículo en Inglés | MEDLINE | ID: mdl-34770394

RESUMEN

Image quality, resolution and scanning time are critical in digital pathology. In order to create a high-resolution digital image, the scanner systems execute stitching algorithms to the digitized images. Due to the heterogeneity of the tissue sample, complex optical path, non-acceptable sample quality or rapid stage movement, the intensities on pictures can be uneven. The evincible and visible intensity distortions can have negative effect on diagnosis and quantitative analysis. Utilizing the common areas of the neighboring field-of-views, we can estimate compensations to eliminate the inhomogeneities. We implemented and validated five different approaches for compensating output images created with an area scanner system. The proposed methods are based on traditional methods such as adaptive histogram matching, regression-based corrections and state-of-the art methods like the background and shading correction (BaSiC) method. The proposed compensation methods are suitable for both brightfield and fluorescent images, and robust enough against dust, bubbles, and optical aberrations. The proposed methods are able to correct not only the fixed-pattern artefacts but the stochastic uneven illumination along the neighboring or above field-of-views utilizing iterative approaches and multi-focal compensations.


Asunto(s)
Procesamiento de Imagen Asistido por Computador , Iluminación , Algoritmos , Artefactos , Cintigrafía
11.
J Chem Ecol ; 46(10): 917-926, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-33026596

RESUMEN

The chemical signatures emitted by fungal substrates are key components for mycophagous insects in the search for food source or for suitable oviposition sites. These volatiles are usually emitted by the fruiting bodies and mycelia. The volatiles attract fungivorous insects, like flowers attract pollinators; certain flowers mimic the shape of mushroom fruiting bodies and even produce a typical mushroom odor to exploit on fungus-insect mutualism. There are numerous insects which are mycophagous or eat fungi additionally, but only a few are considered a threat in agriculture. Lycoriella ingenua is one of the most serious pests in mushroom cultivation worldwide. Here we attempt to examine the role of environmental volatiles upon behavioral oviposition preference. In two-choice bioassays, fungus gnats preferred uncolonized compost compared to colonized compost but preferred colonized compost against nothing. However, when colonized compost was paired against distilled water, no significant choice was observed. The comparison of fresh casing material and mycelium colonized casing material resulted in no significant preference. From colonized compost headspace, three antennally active volatiles were isolated by gas chromatography coupled with electroantennography and subsequently identified with gas chromatography coupled mass spectrometry as 1-hepten-3-ol, 3-octanone and 1-octen-3-ol. In behavioral assays the addition of said synthetic volatiles to uncolonized compost separately and in combination to mimic colonized compost resulted in avoidance. We thus partially elucidate the role of fungal volatiles in the habitat seeking behavior of Lycoriella ingenua.


Asunto(s)
Agaricus/crecimiento & desarrollo , Compostaje , Dípteros/fisiología , Micelio/crecimiento & desarrollo , Percepción Olfatoria/fisiología , Compuestos Orgánicos Volátiles/química , Animales , Conducta Animal/efectos de los fármacos , Señales (Psicología) , Control de Insectos/métodos , Oviposición , Compuestos Orgánicos Volátiles/farmacología
12.
BMC Cancer ; 19(1): 1059, 2019 Nov 06.
Artículo en Inglés | MEDLINE | ID: mdl-31694571

RESUMEN

BACKGROUND: Long non-coding RNAs (lncRNAs) play a fundamental role in colorectal cancer (CRC) development, however, lncRNA expression profiles in CRC and its precancerous stages remain to be explored. We aimed to study whole genomic lncRNA expression patterns in colorectal adenoma-carcinoma transition and to analyze the underlying functional interactions of aberrantly expressed lncRNAs. METHODS: LncRNA expression levels of colonic biopsy samples (20 CRCs, 20 adenomas (Ad), 20 healthy controls (N)) were analyzed with Human Transcriptome Array (HTA) 2.0. Expression of a subset of candidates was verified by qRT-PCR and in situ hybridization (ISH) analyses. Furthermore, in silico validation was performed on an independent HTA 2.0, on HGU133Plus 2.0 array data and on the TCGA COAD dataset. MiRNA targets of lncRNAs were predicted with miRCODE and lncBase v2 algorithms and miRNA expression was analyzed on miRNA3.0 Array data. MiRNA-mRNA target prediction was performed using miRWALK and c-Met protein levels were analyzed by immunohistochemistry. Comprehensive lncRNA-mRNA-miRNA co-expression pattern analysis was also performed. RESULTS: Based on our HTA results, a subset of literature-based CRC-associated lncRNAs showed remarkable expression changes already in precancerous colonic lesions. In both Ad vs. normal and CRC vs. normal comparisons 16 lncRNAs, including downregulated LINC02023, MEG8, AC092834.1, and upregulated CCAT1, CASC19 were identified showing differential expression during early carcinogenesis that persisted until CRC formation (FDR-adjusted p < 0.05). The intersection of CRC vs. N and CRC vs. Ad comparisons defines lncRNAs characteristic of malignancy in colonic tumors, where significant downregulation of LINC01752 and overexpression of UCA1 and PCAT1 were found. Two candidates with the greatest increase in expression in the adenoma-carcinoma transition were further confirmed by qRT-PCR (UCA1, CCAT1) and by ISH (UCA1). In line with aberrant expression of certain lncRNAs in tumors, the expression of miRNA and mRNA targets showed systematic alterations. For example, UCA1 upregulation in CRC samples occurred in parallel with hsa-miR-1 downregulation, accompanied by c-Met target mRNA overexpression (p < 0.05). CONCLUSION: The defined lncRNA sets may have a regulatory role in the colorectal adenoma-carcinoma transition. A subset of CRC-associated lncRNAs showed significantly differential expression in precancerous samples, raising the possibility of developing adenoma-specific markers for early detection of colonic lesions.


Asunto(s)
Adenoma/genética , Carcinoma/genética , Neoplasias Colorrectales/genética , Perfilación de la Expresión Génica/métodos , Regulación Neoplásica de la Expresión Génica , ARN Largo no Codificante/genética , Adenoma/patología , Adulto , Anciano , Carcinoma/patología , Neoplasias Colorrectales/patología , Ontología de Genes , Redes Reguladoras de Genes , Humanos , Persona de Mediana Edad , Modelos Genéticos , Adulto Joven
13.
J Chem Ecol ; 45(8): 657-666, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31407199

RESUMEN

The box tree moth, Cydalima perspectalis, is an invasive pest in Europe causing damage on Buxus species. In this study, we aimed to develop a "bisexual" lure to attract both female and male moths. Based on a previous screening bioassay we tested methyl salicylate, phenylacetaldehyde and eugenol as potential attractants in different combinations. The trapping results showed that both binary and ternary blends attracted male and female moths. Catches with these blends were comparable to catches with the synthetic pheromone. Subsequently we carried out single sensillum recordings, which proved the peripheral detection of the above-mentioned compounds on male and female antennae. To identify synergistic flower volatiles, which can be also attractive and can increase the trap capture, we collected flower headspace volatiles from 12 different flowering plant species. Several components of the floral scents evoked good responses from antennae of both females and males in gas chromatography-electroantennographic detection. The most active components were tentatively identified by gas chromatography coupled mass spectrometry as benzaldehyde, cis-ß-ocimene, (±)-linalool and phenethyl alcohol. These selected compounds in combination did not increase significantly the trap capture compared to the methyl salicylate- phenyacetaldehyde blend. Based on these results we discovered the first attractive blend, which was able to attract both adult male and female C. perspectalis in field conditions. These results will yield a good basis for the optimization and development of a practically usable bisexual lure against this invasive pest.


Asunto(s)
Conducta Animal/efectos de los fármacos , Mariposas Nocturnas/fisiología , Feromonas/farmacología , Monoterpenos Acíclicos , Animales , Eugenol/química , Eugenol/farmacología , Femenino , Flores/química , Flores/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Control de Insectos , Masculino , Monoterpenos/química , Monoterpenos/farmacología , Feromonas/análisis , Robinia/química , Robinia/metabolismo , Rosa/química , Rosa/metabolismo , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/farmacología
14.
BMC Cancer ; 18(1): 695, 2018 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-29945573

RESUMEN

BACKGROUND: DNA mutations occur randomly and sporadically in growth-related genes, mostly on cytosines. Demethylation of cytosines may lead to genetic instability through spontaneous deamination. Aims were whole genome methylation and targeted mutation analysis of colorectal cancer (CRC)-related genes and mRNA expression analysis of TP53 pathway genes. METHODS: Long interspersed nuclear element-1 (LINE-1) BS-PCR followed by pyrosequencing was performed for the estimation of global DNA metlyation levels along the colorectal normal-adenoma-carcinoma sequence. Methyl capture sequencing was done on 6 normal adjacent (NAT), 15 adenomatous (AD) and 9 CRC tissues. Overall quantitative methylation analysis, selection of top hyper/hypomethylated genes, methylation analysis on mutation regions and TP53 pathway gene promoters were performed. Mutations of 12 CRC-related genes (APC, BRAF, CTNNB1, EGFR, FBXW7, KRAS, NRAS, MSH6, PIK3CA, SMAD2, SMAD4, TP53) were evaluated. mRNA expression of TP53 pathway genes was also analyzed. RESULTS: According to the LINE-1 methylation results, overall hypomethylation was observed along the normal-adenoma-carcinoma sequence. Within top50 differential methylated regions (DMRs), in AD-N comparison TP73, NGFR, PDGFRA genes were hypermethylated, FMN1, SLC16A7 genes were hypomethylated. In CRC-N comparison DKK2, SDC2, SOX1 genes showed hypermethylation, while ERBB4, CREB5, CNTN1 genes were hypomethylated. In certain mutation hot spot regions significant DNA methylation alterations were detected. The TP53 gene body was addressed by hypermethylation in adenomas. APC, TP53 and KRAS mutations were found in 30, 15, 21% of adenomas, and in 29, 53, 29% of CRCs, respectively. mRNA expression changes were observed in several TP53 pathway genes showing promoter methylation alterations. CONCLUSIONS: DNA methylation with consecutive phenotypic effect can be observed in a high number of promoter and gene body regions through CRC development.


Asunto(s)
Neoplasias Colorrectales/genética , Metilación de ADN , Exones , Mutación , Regiones Promotoras Genéticas , Adenoma/genética , Islas de CpG , Humanos , Elementos de Nucleótido Esparcido Largo , Transducción de Señal , Proteína p53 Supresora de Tumor/fisiología
15.
Orv Hetil ; 159(6): 223-233, 2018 Feb.
Artículo en Húngaro | MEDLINE | ID: mdl-29400102

RESUMEN

INTRODUCTION: Cell-free DNA (cfDNA) was first detected in human plasma in the 1940s, but the knowledge on its regulation and rate of release is incomplete. CfDNA can originate from both normal and tumour cells. AIM: Our aims were to investigate the rate of cfDNA's release in SHO mice/HT-29 colorectal adenocarcinoma cell line xenograft model and to define the decay of methylated and non-methylated DNA fragments in C57BL/6 bloodstream. METHOD: SHO mice were xenografted with human HT-29 cells, than blood samples were collected over 2 months. CfDNA was isolated, then quantified by real-time PCR with highly specific genomic and mitochondrial human and mouse primer sets. This method permitted to define the ratio of human/mouse DNA. To assess the degradation rate of cfDNA, 3000 bp sized methylated and non-methylated DNA fragments were injected into healthy and C38 tumour-cell vaccinated C57BL/6 mice's bloodstream. The decay of amplicons was measured with 19 PCR assays. RESULTS: The amount of human DNA until the 2nd week was below the limit of detection. From the third week, a continuous growth was experienced, which reached 18.26% by the 8th week. Moreover, it was found that in healthy animals the non-methylated DNA disappears from the plasma after 6 hours, while the methylated fragment was detectable even after 24 hours. In animals with tumour, both amplicons were detectable after 24 hours. CONCLUSION: The examination of the role and mechanism of cfDNA shows an increasing level of interest. This work can contribute to a better understanding of the release and degradation of cfDNA. Orv Hetil. 2018; 159(6): 223-233.


Asunto(s)
Adenocarcinoma/sangre , Biomarcadores de Tumor/sangre , Ácidos Nucleicos Libres de Células/sangre , Fragmentación del ADN , ADN de Neoplasias/sangre , Animales , Ratones , Ratones Endogámicos C57BL
16.
Orv Hetil ; 159(1): 3-15, 2018 Jan.
Artículo en Húngaro | MEDLINE | ID: mdl-29291647

RESUMEN

Besides the genetic research, increasing number of scientific studies focus on epigenetic phenomena - such as DNA methylation - regulating the expression of genes behind the phenotype, thus can be related to the pathomechanism of several diseases. In this review, we aim to summarize the current knowledge about the evolutionary appearance and functional diversity of DNA methylation as one of the epigenetic mechanisms and to demonstrate its role in aging and cancerous diseases. DNA methylation is also characteristic/also appear to prokaryotes, eukaryotes and viruses. In prokaryotes and viruses, it provides defence mechanisms against extragenous DNA. DNA methylation in prokaryotes plays a significant role in the regulation of transcription, the initiation of replication and in Dam-directed mismatch repair. In viruses, it participates not only in defence mechanisms, but in the assembly of capsids as well which is necessary for spreading. In eukaryotes, DNA methylation is involved in recombination, replication, X chromosome inactivation, transposon control, regulation of chromatin structure and transcription, and it also contributes to the imprinting phenomenon. Besides the above-mentioned aspects, DNA methylation also has an evolutionary role as it can change DNA mutation rate. Global hypomethylation appearing during aging and in cancerous diseases can lead to genetic instablility and spontaneous mutations through its role in the regulation of transposable elements. Local hypermethylated alterations such as hypermethylation of SFRP1, SFRP2, DKK1 and APC gene promoters can cause protein expression changes, thus contribute to development of cancer phenotype. DNA methylation alterations during aging in cancerous diseases support the importance of epigenetic research focusing on disease diagnostics and prognostics. Orv Hetil. 2018; 159(1): 3-15.


Asunto(s)
Envejecimiento/metabolismo , Biomarcadores de Tumor/metabolismo , Epigénesis Genética/genética , Neoplasias/genética , Anciano , Anciano de 80 o más Años , Metilación de ADN , Humanos , Neoplasias/metabolismo
17.
Cytometry A ; 91(6): 595-608, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28472544

RESUMEN

Nuclear estrogen receptor (ER), progesterone receptor (PR) and Ki-67 protein positive tumor cell fractions are semiquantitatively assessed in breast cancer for prognostic and predictive purposes. These biomarkers are usually revealed using immunoperoxidase methods resulting in diverse signal intensity and frequent inhomogeneity in tumor cell nuclei, which are routinely scored and interpreted by a pathologist during conventional light-microscopic examination. In the last decade digital pathology-based whole slide scanning and image analysis algorithms have shown tremendous development to support pathologists in this diagnostic process, which can directly influence patient selection for targeted- and chemotherapy. We have developed an image analysis algorithm optimized for whole slide quantification of nuclear immunostaining signals of ER, PR, and Ki-67 proteins in breast cancers. In this study, we tested the consistency and reliability of this system both in a series of brightfield and DAPI stained fluorescent samples. Our method allows the separation of overlapping cells and signals, reliable detection of vesicular nuclei and background compensation, especially in FISH stained slides. Detection accuracy and the processing speeds were validated in routinely immunostained breast cancer samples of varying reaction intensities and image qualities. Our technique supported automated nuclear signal detection with excellent efficacy: Precision Rate/Positive Predictive Value was 90.23 ± 4.29%, while Recall Rate/Sensitivity was 88.23 ± 4.84%. These factors and average counting speed of our algorithm were compared with two other open source applications (QuPath and CellProfiler) and resulted in 6-7% higher Recall Rate, while 4- to 30-fold higher processing speed. In conclusion, our image analysis algorithm can reliably detect and count nuclear signals in digital whole slides or any selected large areas i.e. hot spots, thus can support pathologists in assessing clinically important nuclear biomarkers with less intra- and interlaboratory bias inherent of empirical scoring. © 2017 International Society for Advancement of Cytometry.


Asunto(s)
Algoritmos , Biomarcadores de Tumor/genética , Neoplasias de la Mama/diagnóstico , Antígeno Ki-67/genética , Receptores de Estrógenos/genética , Receptores de Progesterona/genética , Automatización de Laboratorios , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Núcleo Celular/metabolismo , Núcleo Celular/patología , Núcleo Celular/ultraestructura , Células Epiteliales/metabolismo , Células Epiteliales/patología , Células Epiteliales/ultraestructura , Femenino , Expresión Génica , Humanos , Interpretación de Imagen Asistida por Computador/métodos , Inmunohistoquímica/métodos , Hibridación Fluorescente in Situ , Sensibilidad y Especificidad
18.
Mod Pathol ; 29(8): 928-38, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27150162

RESUMEN

Exosomes are small membrane vesicles that have important roles in transporting a great variety of bioactive molecules between epithelial compartment and their microenvironment during tumor formation including colorectal adenoma-carcinoma sequence. We tested the mRNA expression of the top 25 exosome-related markers based on ExoCharta database in healthy (n=49), adenoma (n=49) and colorectal carcinoma (n=49) patients using Affymetrix HGU133 Plus2.0 microarrays. Most related genes showed significantly elevated expression including PGK1, PKM, ANXA5, ENO1, HSP90AB1 and MSN during adenoma-carcinoma sequence. Surprisingly, the expression of ALIX (ALG 2-interacting protein X), involved in multivesicular body (MVB) and exosome formation, was significantly reduced in normal vs adenoma (P=5.02 × 10(-13)) and in normal vs colorectal carcinoma comparisons (P=1.51 × 10(-10)). ALIX also showed significant reduction (P<0.05) at the in situ protein level in the epithelial compartment of adenoma (n=35) and colorectal carcinoma (n=37) patients compared with 27 healthy individuals. Furthermore, significantly reduced ALIX protein levels were accompanied by their gradual transition from diffuse cytoplasmic expression to granular signals, which fell into the 0.6-2 µm diameter size range of MVBs. These ALIX-positive particles were seen in the tumor nests, including tumor-stroma border, which suggest their exosome function. MVB-like structures were also detected in tumor microenvironment including α-smooth muscle actin-positive stromal cells, budding off cancer cells in the tumor front as well as in cancer cells entrapped within lymphoid vessels. In conclusion, we determined the top aberrantly expressed exosome-associated markers and revealed the transition of diffuse ALIX protein signals into a MVB-like pattern during adenoma-carcinoma sequence. These tumor-associated particles seen both in the carcinoma and the surrounding microenvironment can potentially mediate epithelial-stromal interactions involved in the regulation of tumor growth, metastatic invasion and therapy response.


Asunto(s)
Adenoma/química , Biomarcadores de Tumor/análisis , Proteínas de Unión al Calcio/análisis , Carcinoma/química , Proteínas de Ciclo Celular/análisis , Neoplasias Colorrectales/química , Complejos de Clasificación Endosomal Requeridos para el Transporte/análisis , Exosomas/química , Cuerpos Multivesiculares/química , Adenoma/genética , Adenoma/patología , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Proteínas de Unión al Calcio/genética , Carcinoma/genética , Carcinoma/patología , Estudios de Casos y Controles , Proteínas de Ciclo Celular/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Complejos de Clasificación Endosomal Requeridos para el Transporte/genética , Exosomas/genética , Exosomas/patología , Femenino , Perfilación de la Expresión Génica/métodos , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Cuerpos Multivesiculares/genética , Cuerpos Multivesiculares/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Microambiente Tumoral
19.
Orv Hetil ; 157(28): 1117-25, 2016 Jul.
Artículo en Húngaro | MEDLINE | ID: mdl-27397424

RESUMEN

INTRODUCTION: Screening, prevention and treatment of familial breast cancer require a multidisciplinary approach. New guidelines were published in the United Kingdom for the management of familial breast cancer. AIM: The authors summarise these new guidelines and analyse the relevant practice in Hungary. METHOD: Relevant guidelines of the National Institute for Health and Care Excellence and Familial Breast Cancer Report (NHS Scotland) are described. RESULTS: New guidelines will increase the number of genetic tests as well as genetic counselling. An increase in the number of breast magnetic resonance imaging is expected, too. Chemoprevention can be offered for individuals with medium risk and above. Promising trials are underway with platinum based chemotherapy and polyADP-ribose polimerase inhibitors for the systemic treatment of familial breast cancer. The increase in the number of genetic tests, counselling, and breast magnetic resonance imaging may have a significant impact on health care budget. CONCLUSIONS: These guidelines will change some aspects of the current management of familial breast cancer. Orv. Hetil., 2016, 157(28), 1117-1125.


Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/genética , Neoplasias de la Mama/terapia , Pruebas Genéticas , Mastectomía Profiláctica , Protocolos de Quimioterapia Combinada Antineoplásica/economía , Proteína BRCA1/genética , Proteína BRCA2/genética , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/prevención & control , Quimioprevención/economía , Quimioprevención/métodos , Quimioprevención/tendencias , Inglaterra , Femenino , Predisposición Genética a la Enfermedad , Pruebas Genéticas/economía , Pruebas Genéticas/normas , Pruebas Genéticas/tendencias , Humanos , Hungría , Internacionalidad , Mamografía , Mutación , Ovariectomía , Guías de Práctica Clínica como Asunto , Mastectomía Profiláctica/economía , Medición de Riesgo , Factores de Riesgo , Salpingectomía , Gales
20.
Orv Hetil ; 157(10): 392-5, 2016 Mar 06.
Artículo en Húngaro | MEDLINE | ID: mdl-26920330

RESUMEN

Leiomyoma is a rare, smooth muscle tumour that can occur everywhere in the human body. The authors present the history of a 60-year-old female, who had a giant, Mullerian type myxoid leiomyoma in the inguinal region mimicking acute abdominal symptoms. After examination the authors removed the soft tissue mass in the right femoral region reaching down in supine position to the middle third of the leg measuring 335 × 495 × 437 mm in greatest diameters in weight 33 kg. Reconstruction of the tissue defect was performed using oncoplastic guidelines. During the follow-up time no tumour recurrence was detected and the quality of life of the patient improved significantly.


Asunto(s)
Leiomioma/diagnóstico , Leiomioma/cirugía , Mixoma/diagnóstico , Mixoma/cirugía , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/cirugía , Diagnóstico Diferencial , Femenino , Hernia Inguinal/diagnóstico , Humanos , Leiomioma/diagnóstico por imagen , Leiomioma/patología , Persona de Mediana Edad , Mixoma/diagnóstico por imagen , Calidad de Vida , Procedimientos de Cirugía Plástica/métodos , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Neoplasias Uterinas/diagnóstico por imagen
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