Multiplex mRNA profiling for the identification of body fluids

Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification.

Vascular calcification may occur at different areas of the vessel wall, including the intima in atherosclerosis and the media in Monckeberg's sclerosis Medial calcification of arteries is common in patients with diabetes mellitus or chronic renal failure Osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand are essential modulators of bone homeostasis and may be involved in the process of vascular calcification In this study we investigated arteries from patients with Monckeberg's sclerosis and atherosclerosis Apoptosis, which precedes vascular calcification in vitro, was assessed by an in situ ligation assay and was localized to the medial layer of arteries (Monckeberg's sclerosis) and the neointima (atherosclerosis) Immunohistochemistry and in situ hybridization revealed OPG immunoreactivity and mRNA expression surrounding calcified areas in the medial layer (Monckeberg's sclerosis), whereas OPG was mainly expressed adjacent to calcified neointimal lesions (atherosclerosis) Receptor activator of nuclear factor-kappaB ligand protein and mRNA were barely or not detectable Of note, TNF-related apoptosis-inducing ligand, an inducer of apoptosis that is also blocked by OPG, displayed a similar spatial distribution as OPG In summary, we demonstrate enhanced apoptosis adjacent to vascular calcification, and the concurrent expression of regulators of apoptosis and osteoclastic differentiation, TNF-related apoptosis-inducing ligand and OPG, suggesting their involvement in the pathogenesis of vascular calcification

Localization of Osteoprotegerin, Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand, and Receptor Activator of Nuclear Factor-κB Ligand in Mönckeberg’s Sclerosis and Atherosclerosis

RNA expression patterns, including the presence and relative abundance of particular mRNA species, provide cell and tissue specific information that could be used for body fluid identification. In this report, we address perceived concerns on the stability, and hence recoverability, of RNA in forensic samples. Stains were prepared from blood, saliva, semen, and vaginal secretions and exposed to a range of environmental conditions from 1 to 547 days. The persistence and stability of RNA within each type of body fluid stain were determined by quantitation of total RNA, and reverse transcriptase-polymerase chain reaction (RT-PCR) using eight different mRNA transcripts from selected housekeeping and tissue-specific genes. The results demonstrate that RNA can be recovered from biological stains in sufficient quantity and quality for mRNA analysis. On average, several hundred nanograms of total RNA was recovered from 50-microL-sized blood and saliva stains, 1 microg from a 50-microL semen stain and nearly 70 microg from a whole vaginal swab. Messenger RNA is detectable in some samples stored at room temperature for at least 547 days. The environmental samples that were protected from direct rain impact exhibited housekeeping and tissue specific mRNA recoverability up to 7 days (saliva and semen), 30 days (blood), or 180 days (vaginal swab). Additionally, rain had a detrimental effect on the recoverability of blood (3 days), saliva (1 day), semen (7 days), and vaginal secretions (3 days) specific transcripts, with one of the mRNA species (the semen marker PRM2) not being detectable after 1 day.

Recovery and Stability of RNA in Vaginal Swabs and Blood, Semen, and Saliva Stains

Background: Diabetic nephropathy (DN) is a serious complication of diabetes mellitus and a common cause of end-stage renal disease. Autophagy has a defensive role against kidney damage caused by hyperglycemia. Mesenchymal stem cell (MSC)-derived exosomes are currently considered as a new promising therapy for chronic renal injury. However, the renal-protective mechanism of exosomes on DN is not completely understood. We examined the potential role of MSC-derived exosomes for enhancement of autophagy activity and their effect on DN. In our study, we used five groups of rats: control; DN; DN treated with exosomes; DN treated with 3-methyladenine (3-MA) and chloroquine (inhibitors of autophagy); and DN treated with 3-methyladenine (3-MA), chloroquine, and exosome groups. We assessed renal function, morphology, and fibrosis. Moreover, ratios of the autophagy markers mechanistic target of rapamycin (mTOR), Beclin-1, light chain-3 (LC3-II), and LC3-II/LC3-I were detected. Additionally, electron microscopy was used for detection of autophagosomes. Results: Exosomes markedly improved renal function and showed histological restoration of renal tissues, with significant increase of LC3 and Beclin-1, and significant decrease of mTOR and fibrotic marker expression in renal tissue. All previous effects were partially abolished by the autophagy inhibitors chloroquine and 3-MA. Conclusion: We conclude that autophagy induction by exosomes could attenuate DN in a rat model of streptozotocin-induced diabetes mellitus.

Mesenchymal Stem Cell-Derived Exosomes Ameliorated Diabetic Nephropathy by Autophagy Induction through the mTOR Signaling Pathway.

https://www.future-science.com/doi/pdf/10.2144/00292bm11

Irreversible heat inactivation of DNase I without RNA degradation.

PCR-based cloning of candidate disease resistance genes from banana (Musa acuminata).

La presente invention concerne des sequences de promoteur qui sont utiles pour moduler la transcription d'un polynucleotide souhaite et/ou dans la construction de genes recombinants pour la transformation de plantes afin d'activer l'expression de sequences de codage etrangeres ou endogenes dans des plantes. Les promoteurs sont fonctionnels a la fois dans des especes de plantes monocotyledones et dicotyledones et sont actifs dans une large gamme d'organes vegetaux en phase vegetative ou generative. L'invention concerne egalement une construction polynucleotidique comprenant un promoteur de l'invention lie de facon operationnelle a un polynucleotide etranger ou endogene codant pour une proteine d'interet ou un transcrit capable de moduler l'expression d'un gene cible. L'invention concerne en outre des cellules vegetales transformees ainsi que des plantes differenciees, des parties vegetales, des tissus vegetaux ou des graines de plantes contenant la construction.

Ilse Wiame

Papers

Irreversible heat inactivation of DNase I without RNA degradation.

PCR-based cloning of candidate disease resistance genes from banana (Musa acuminata).

Promoteurs de banane