The use of biologic therapies for the management of knee osteoarthritis (OA) has largely increased in recent years. The purpose of this study was to evaluate the efficiency and the therapeutic potential of platelet-rich plasma (PRP) and autologous adipose tissue (AAT) injections as a treatment for knee OA. Sixty participants were enrolled in the study: 20 healthy ones and 40 with minimal to moderate knee OA (KL I-III). The OA patients were randomly assigned either to the PRP or to the AAT group. The PRP samples showed a low expression level of NF-κB-responsive gene CCL5 and high expression levels of classic inflammatory and TNF-l INF responses. The AAT injection product was prepared using a Lipogems device, and its regenerative potential as well as the ability for expansion of mesenchymal stem cells were tested in the cell culture conditions. The patient assessments were carried out five times. Significant improvement was observed regardless of the treatment method in the VAS, KOOS, WOMAC and IKDC 2000 subjective evaluations as well as in the functional parameters. Intra-articular injections of AAT or PRP improved pain, symptoms, quality of life and functional capacity with a comparable effectiveness in the patients with mild to moderate knee osteoarthritis.

For a very long time, RNA molecules were treated as transistory molecules, by which the genetic information flows from DNA to proteins; the model proposed in the 1960s accepted that proteins are both the products and the regulators of gene expression. Since then, thousands of reports proved that RNAs should be thought about as the factors that do control gene expression. The pervasive transcription has been reported in many eukaryotic organisms, illustrating a highly interwoven transcriptome organization that includes hundreds of previously unknown noncoding RNAs. The key roles of noncoding RNAs (microRNAs and small interfering RNAs) in gene expression regulation are no longer surprising, as are new classes of noncoding RNAs constantly being discovered. Transfer RNAs (tRNAs) are the second most abundant type of RNAs in the cell. Advances in high‐throughput sequencing technologies exposed the existence of functional, regulatory tRNA‐derived RNA fragments (tRFs), generated from precursor and mature tRNAs. These tRF molecules have been found to play central roles during stress and different pathological conditions. Herein, we present the critical assessment of the discoveries made in the field of tRNA‐derived fragments in the past 15 years in various pathogenic and nonpathogenic yeast species.

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Chemical probing, for decades, has been one of the most popular tools for studying the secondary structure of RNA molecules. Recently, protocols for simultaneous analysis of multiple RNAs have been developed, enabling in vivo transcriptome-wide interrogation of the RNA structure dynamics. One of the most popular methods is the selective 2′-hydroxyl acylation analyzed by primer extension and mutational profiling (SHAPE-MaP). In this study, we describe the evaluation of this protocol by addressing the influence of the reverse transcription enzymes, buffer conditions, and chemical probes on the properties of the cDNA library and the quality of mutational profiling-derived structural signals. Our results reveal a SuperScript IV (SSIV) reverse transcriptase as a more efficient enzyme for mutational profiling of SHAPE adducts and shed new light on the role of Mn2+ cations in the modulation of SSIV readthrough efficiency.

Regulation of gene expression by premature termination of transcription has been well described in all domains of life, including metazoans, yeast, plants, and bacteria. Although methods for identification of such regulatory events by sequencing are available, the focused biochemical studies of the mechanism are hampered by lack of highly sensitive and accurate experimental methods. Here, we propose a new method for absolute quantification of premature transcription termination events, PTT-quant. It is based on highly sensitive two-step digital droplet PCR protocol, coupled with normalized cDNA synthesis attained by site-specific pre-cleavage of investigated transcripts with RNase H. As a consequence, our method enables the reliable and sensitive quantification of both, prematurely terminated and full-length transcripts. By application of our method to investigation of transcriptional riboswitches in Bacillus subtilis, we were able to precisely measure the dynamics of S-adenosylmethionine (SAM) riboswitch induction, which turned to be ~ 23% higher in comparison the results obtained without cDNA synthesis normalization.Key points• A novel method for quantification of premature transcription termination events was established.• PTT-quant measures absolute concentration of full-length and terminated transcripts.• RNase H and the digital droplet PCR technique is used in PTT-quant.