Effects of mesenchymal stromal cells versus serum on tendon healing in a controlled experimental trial in an equine model

Abstract Background Mesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone. Methods Quadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression. Results Tendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24. Conclusions These data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine

PCR multiplexovani pomoci jedineho fluorescenčního kanálu pro detekovani viru ptačí chřipky pomocí kvantitativní PCR s interkalačním bavivem

Since its invention in 1985 the polymerase chain reaction (PCR) has become a well-established method for amplification and detection of segments of double-stranded DNA. Incorporation of fluorogenic probe or DNA intercalating dyes (such as SYBR Green) into the PCR mixture allowed real-time reaction monitoring and extraction of quantitative information (qPCR). Probes with different excitation spectra enable multiplex qPCR of several DNA segments using multi-channel optical detection systems. Here we show multiplex qPCR using an economical EvaGreen-based system with single optical channel detection. Previously reported non quantitative multiplex realtime PCR techniques based on intercalating dyes were conducted once the PCR is completed by performing melting curve analysis (MCA). The technique presented in this paper is both qualitative and quantitative as it provides information about the presence of multiple DNA strands as well as the number of starting copies in the tested sample. Besides important internal control, multiplex qPCR also allows detecting concentrations of more than one DNA strand within the same sample. Detection of the avian influenza virus H7N9 by PCR is a well established method. Multiplex qPCR greatly enhances its specificity as it is capable of distinguishing both haemagglutinin (HA) and neuraminidase (NA) genes as well as their ratio.Od svého vynálezu v 1985 polymerázová řetězová reakce (PCR) se stal dobře zavedená metoda pro amplifikaci a detekci segmentů dvouřetězcové DNA. Začlenění barviva fluorogenní sondy nebo DNA interkalační (jako SYBR Green) do PCR směsi povoleno Monitorování v reálném čase a extrakce kvantitativních informací (qPCR) reakce. Sondy s různé excitační spektra umožňují multiplexní qPCR experimentu z několika segmentů DNA s využitím multi-kanálové optické systémy detekce. Zde vám ukážeme, multiplex qPCR experimentu pomocí ekonomické EvaGreen bázi Systém s jedním detekcí optickou kanálu. Dříve hlášeny non kvantitativní PCR techniky multiplex v reálném čase založené na interkalační barviva byly provedeny po PCR je dokončen provádění analýzy křivky tání (MCA). Technika uvedené v tomto dokumentu je jak kvalitativní a kvantitativní, neboť poskytuje informace o přítomnosti několika vláken DNA, jakož i počet spuštění kopií v testovaném vzorku. Kromě významnou vnitřní kontroly, multiplexní qPCR také umožňuje detekci koncentrace více než jednoho vlákna DNA ve stejném vzorku. Detekce viru ptačí chřipky H7N9 pomocí PCR je dobře zavedená metoda. Multiplex qPCR výrazně zvyšuje své specifičnosti jako je schopen rozlišit jak hemaglutinin (HA) a neuraminidázou (NA) genů, stejně jako jejich poměr

Droplet-based synthesis of homogeneous magnetic iron oxide nanoparticles

Nanoparticles have gained large interest in a number of different fields due to their unique properties. In medical applications, for example, magnetic nanoparticles can be used for targeting, imaging, magnetically induced thermotherapy, or for any combination of the three. However, it is still a challenge to obtain narrowly dispersed, reproducible particles through a typical lab-scale synthesis when researching these materials. Here, we present a droplet capillary reactor that can be used for the synthesis of magnetic iron oxide nanoparticles. Compared to conventional batch synthesis, the particles synthesized in our droplet reactor have a narrower size distribution and a higher reproducibility. Furthermore, we demonstrate how the particle size can be changed from 5.2 ± 0.9 nm to 11.8 ± 1.7 nm by changing the reaction temperature and droplet residence time in the droplet capillary reactor

Palm-Sized Device for Point-of-Care Ebola Detection

We show the utilization of a recently developed cellphone-sized real-time polymerase chain reaction (PCR) device to detect Ebola virus RNA using single-step reverse transcription PCR (RT-PCR). The device was shown to concurrently perform four PCRs, each with a sample volume of 100 nL: one positive control with both Ebola and GAPDH RNA and one negative control. The last two positions were used to measure the GAPDH and the Ebola content of a sample. A comparison of threshold cycles (C_T) from the two samples provided relative quantification. The entire process, which consisted of reverse transcription, PCR amplification, and melting curve analysis (MCA), was conducted in less than 37 min. The next step will be integration with a sample preparation unit to form an integrated sample-to-answer system for point-of-care infectious disease diagnostics