The epithelium that lines the epididymal duct establishes optimal milieu in which spermatozoa mature, acquire motility, and are stored. This finely tuned environment also protects antigenic sperm against pathogens autoimmunity, potential causes of transient or permanent infertility. is pseudostratified contains basal cells (BCs) located beneath other epithelial cells. Previous studies showed mouse epididymis, BCs possess macrophage-like characteristics. However, we previously identified a...

Extracellular vesicles (EVs) are gaining interest as efficient, biocompatible vehicles for cellular delivery of therapeutic cargo. Precipitation-based methods the isolation EVs remain popular due to ease use and lack requirements specialized equipment. We describe here a novel charge-based EV method that is simple, scalable, uses inexpensive polyethylenimine (PEI) polymers. GFP-expressing were isolated from conditioned cell culture (CCM) media HEK293-GFP cells using either branched 10 kDa...

INTRODUCTION: High-grade gliomas (HGG) are universally fatal (with a progression-free survival of 9 months and overall 14 in the Nova Scotia cohort). Extracellular vesicles (EVs) released into plasma from tumor microenvironment (TME) can contribute to pro-tumorigenic setting. Understanding contents these EVs may further our understanding HGG potentially reveal novel targets biomarkers. tRNA fragments among most abundant small RNAs contained have recently been described as having pro-tumour...

A subset of basal cells (BCs) in the initial segment (IS) mouse epididymis has a slender body projection between adjacent epithelial cells. We show here that these projections occasionally cross apical tight junctions and are contact with luminal environment. Luminal testicular factors critical for establishment IS epithelium, we investigated their role regulation this sensing property. Efferent duct ligation (EDL) was performed to block flow from testis without affecting blood flow....

Abstract Several studies have demonstrated the potential uses of extracellular vesicles (EVs) for liquid biopsy‐based diagnostic tests and therapeutic applications; however, clinical use EVs presents a challenge as many currently‐available EV isolation methods limitations related to efficiency, purity, complexity methods. Moreover, do not perform efficiently in all biofluids due their differential physicochemical properties. Thus, there continues be need novel that are simple, robust,...

Abstract The initial segment (IS) of the epididymis plays an essential role in male fertility. IS epithelium is undifferentiated and nonfunctional at birth. Prior to puberty, undergoes differentiation that leads formation a fully functional organ. However, mechanistic details this program are not well understood. To explore further, we used genetic engineering create kinase dead allele ROS1 receptor tyrosine mice studied effects activity on epithelium. We show expression activation coincides...

Pancreatic ductal adenocarcinoma (PDAC) has a high fatality rate, mainly due to its asymptomatic nature until late-stage disease and therefore delayed diagnosis that leads lack of timely treatment intervention. Consequently, there is significant need for better methods screen populations are at risk developing PDAC. Such advances would result in earlier diagnosis, more options, ultimately outcomes patients. Several recent studies have applied the concept liquid biopsy, which sampling...

Resistin is a new adipocytokine which expressed in rat, mouse and possibly human adipose tissue. Its putative role as mediator of insulin resistance controversial. We hypothesized that resistin, like leptin, would have multiple roles non-adipose tissues we reported resistin brain pituitary. Moreover, expression female pituitary developmentally regulated maximal occurs peripubertally. Although the endogenous has not been determined, our data suggest could be important postnatal maturation...

Abstract Epithelial cells are generally considered to be static relative their neighbours. Basal in pseudostratified epithelia display a single long cytoplasmic process that can cross the tight junction barrier reach lumen. Using vivo microscopy visualize epididymis, model system for study of epithelia, we report here surprising discovery these basal cell projections—which call axiopodia—periodically extend and retract over time. We found axiopodia extensions retractions follow an...

Clear cells express the vacuolar proton-pumping H + -ATPase (V-ATPase) and acidify lumen of epididymis, a process that is essential for male fertility. The renin-angiotensin-aldosterone system (RAAS) regulates fluid electrolyte balance in previous study showed binding aldosterone exclusively to epididymal clear (Hinton BT, Keefer DA. Steroid Biochem 23: 231–233, 1985). We examined here role regulation V-ATPase epididymis. RT-PCR expression mineralocorticoid receptor [MR; nuclear subfamily 3,...

Extracellular vesicles (EVs) have been recognized as a rich material for the analysis of DNA, RNA, and protein biomarkers. A remaining challenge deployment EV-based diagnostic prognostic assays in liquid biopsy testing is development an EV isolation method that amenable to clinical lab setting compatible with multiple types biomarker analyses. We previously designed synthetic peptide, known Vn96 (ME kit), which efficiently isolates EVs from biofluids short timeframe without use specialized...

RNA sequencing analysis is an important field in the study of extracellular vesicles (EVs), as these particles contain a variety species that may have diagnostic, prognostic and predictive value. Many bioinformatics tools currently used to analyze EV cargo rely on third-party annotations. Recently, unannotated expressed RNAs has become interest, since provide complementary information traditional annotated biomarkers or help refine biological signatures machine learning by including unknown...

K(+) channels play a role in the proliferation of cancer cells. We have investigated effects specific channel inhibitors on basal and oestrogen-stimulated breast cells.Using mammary adenocarcinoma cell line MCF-7 we assayed by radiolabelled thymidine incorporation absence or presence various with without 17beta-oestradiol.Inhibitors K(v)10.1 K(Ca)3.1 suppressed cells, but not proliferation. TRAM-34, inhibitor increased decreased depending concentration. At intermediate concentrations (3-10...

Background: High-grade gliomas (HGG) present challenges with short post-surgery survival and high progression rates. Extracellular vesicles (EVs) in the tumor microenvironment (TME) contribute to a pro-tumorigenic setting. Investigating Transfer RNA fragments (TfRNA) HGG patient plasma EVs reveals potential biomarkers therapeutic targets, shedding light on molecular landscape for enhanced diagnostic strategies. This study examines TfRNA 10 patients at diagnosis, offering insights into...

<title>Abstract</title> <bold>Background: </bold>High grade gliomas (HGGs) and cells of the tumour microenvironment (TME) secrete extracellular vesicles(EVs) into plasma that contain genetic protein cargo, which function in paracrine signaling. Isolation these EVs their cargo from could lead to a simplistic tool can inform on diagnosis disease course HGG. <bold>Methods: </bold>In present study, were captured utilizing peptide affinity method (Vn96 peptide) HGG patients normal controls...

Abstract Transfer RNA (tRNA) are non-coding RNAs that bring amino acids to ribosomes during translation. Recent studies have shown cleaved fragments derived from tRNA (tRFs) serve regulatory roles in epigenetics, signaling, transcription, and tRFs can be released cells found extracellular vesicles perform some aspects of paracrine signaling. Utilizing high throughput small sequencing, we identified over 750 uniquely or multi-mapped (6.9% total sRNA mapped reads) high-grade glioma (IDH-WT)...

The common northern sea star Asterias vulgaris is exposed to seasonal variation in temperature from -2 17°C. A. exhibits an annual reproductive cycle, i.e., the testes increase slowly size during fall and winter, reach maximal early spring. Slow testicular growth winter has been attributed low field temperatures. Previous studies indicate that specific activity of ornithine decarboxylase levels polyamines putrescine, spermidine, spermine decrease mid-winter spring, coincident with changes...

Epididymal basal cells (BCs) sense the luminal environment by sending cell body projections that cross tight‐junction barrier. BCs express EGF receptor and its associated tyrosine kinase, c‐ Src, we examined their role in sensing ability. were visualized vivo multiphoton microscopy our newly generated transgenic mice (KR5‐tdTomato), which tdTomato exclusively BCs. We found periodically elongate retract luminal‐reaching projections. Laser scanning confocal followed 3D modeling showed...

Basal cells (BCs) extend slender body projections that can cross the tight‐junction barrier to scan luminal environment of mouse proximal epididymis. We studied here role testicular factors in regulation this sensing property. Efferent duct ligation (EDL) was performed block flow from testis without affecting blood flow. Keratin 5 (K5) labeling showed number BCs with a luminal‐reaching projection significantly reduced time dependent manner 1 day days after EDL, compared controls. Double for...