Microorganisms, regardless of whether pathogenic or not, may cause enormous economic losses due to adverse effects on human and animal health, or by damaging the quality of the agricultural and food products. Based on...Microorganisms, regardless of whether pathogenic or not, may cause enormous economic losses due to adverse effects on human and animal health, or by damaging the quality of the agricultural and food products. Based on these effects, the development of prompt molecular methods and their involvement in the practical pathogen diagnostic diagnostics is more than actual. This paper is focused on the evaluation of easy-to-perform and highly budget-friendly, PCR-related DNA purification protocols for diagnostic purposes especially in water or similar simple matrices. The slight modifications of earlier described DNA isolation methods, which rely on chelate exchange resin and/or ethanol-sodium-based heat lysis, we reevaluated in comparison with a widely used commercial kit. The efficiency of DNA purification techniques was assessed from Gramnegative as well as Gram-positive bacteria and yeast using quantitative PCR. The effectivity of different methods tested may vary depending on the bacterial or yeast species in question. Nevertheless, in our hands, the chelate exchange resin-based methods were found to be the most robust and/or satisfying at least by an acceptable reproducibility rate. Our presented results support the potential of low-cost but still sensitive molecular microbe detection procedures consisting of only a few pipetting steps resulting in good reproducibility and the least possible environmental burden, serving as a good starting point for developments of matrix-specific processes and methods.展开更多
MicroRNAs (miRNAs) are posttranscriptional regulators fine-tuning the level of most messenger RNAs (mRNAs) and proteins in mammalian cells. Their expression is dysregulated in neoplastic cells and upregulated or downr...MicroRNAs (miRNAs) are posttranscriptional regulators fine-tuning the level of most messenger RNAs (mRNAs) and proteins in mammalian cells. Their expression is dysregulated in neoplastic cells and upregulated or downregulated miRNAs play an important role in tumorigenesis. Changes in the miRNA transcriptome appear to be suitable markers for the differential diagnosis of various B-cell lymphoma types and there are therapeutic attempts to normalize the expression level of key cellular miRNAs involved in lymphomagenesis. In this review we wish to outline the most recent developments in the application of sophisticated, chemically modified antisense oligonucleotides and their nanoparticle complexes to suppress oncogenic miRNAs. These advances form the basis of a new therapeutic approach that may complement current protocols for B-cell lymphoma therapy. Anti-cytokine therapy aiming at the removal of cytokines that activate key oncomirs, and switching on silenced tumor suppressor miRNAs by epigenetic drugs might also be considered, on the long run, in the treatment of well defined B-cell lymphoma types.展开更多
文摘Microorganisms, regardless of whether pathogenic or not, may cause enormous economic losses due to adverse effects on human and animal health, or by damaging the quality of the agricultural and food products. Based on these effects, the development of prompt molecular methods and their involvement in the practical pathogen diagnostic diagnostics is more than actual. This paper is focused on the evaluation of easy-to-perform and highly budget-friendly, PCR-related DNA purification protocols for diagnostic purposes especially in water or similar simple matrices. The slight modifications of earlier described DNA isolation methods, which rely on chelate exchange resin and/or ethanol-sodium-based heat lysis, we reevaluated in comparison with a widely used commercial kit. The efficiency of DNA purification techniques was assessed from Gramnegative as well as Gram-positive bacteria and yeast using quantitative PCR. The effectivity of different methods tested may vary depending on the bacterial or yeast species in question. Nevertheless, in our hands, the chelate exchange resin-based methods were found to be the most robust and/or satisfying at least by an acceptable reproducibility rate. Our presented results support the potential of low-cost but still sensitive molecular microbe detection procedures consisting of only a few pipetting steps resulting in good reproducibility and the least possible environmental burden, serving as a good starting point for developments of matrix-specific processes and methods.
文摘MicroRNAs (miRNAs) are posttranscriptional regulators fine-tuning the level of most messenger RNAs (mRNAs) and proteins in mammalian cells. Their expression is dysregulated in neoplastic cells and upregulated or downregulated miRNAs play an important role in tumorigenesis. Changes in the miRNA transcriptome appear to be suitable markers for the differential diagnosis of various B-cell lymphoma types and there are therapeutic attempts to normalize the expression level of key cellular miRNAs involved in lymphomagenesis. In this review we wish to outline the most recent developments in the application of sophisticated, chemically modified antisense oligonucleotides and their nanoparticle complexes to suppress oncogenic miRNAs. These advances form the basis of a new therapeutic approach that may complement current protocols for B-cell lymphoma therapy. Anti-cytokine therapy aiming at the removal of cytokines that activate key oncomirs, and switching on silenced tumor suppressor miRNAs by epigenetic drugs might also be considered, on the long run, in the treatment of well defined B-cell lymphoma types.
