Immunofluorescence characterization of innervation and nerve-immune cell interactions in mouse lymph nodes
The peripheral nervous system communicates specifically with the immune system via local interactions. These interactions include the “hardwiring” of sympathetic/parasympathetic (efferent) and sensory nerves (afferent) to primary (e.g., thymus and bone marrow) and secondary (e.g., lymph node, spleen, and gut-associated lymphoid tissue) lymphoid tissue/organs. To gain a better understanding of this bidirectional interaction/crosstalk between the two systems, we have investigated the distribution of nerve fibres and PNS-immune cell associationsin situ in the mouse lymph node by using immunofluorescent staining and confoc...
Source: European Journal of Histochemistry - October 18, 2019 Category: Biomedical Science Source Type: research
Digital PCR - Methods and Protocols
Sensitivity, reproducibility, precision and accuracy are recurring words in this book. Indeed, digital PCR (dPCR) represents a major step forward in the quantification of nucleic acids. In this field, for many years it has been used the quantitative PCR (qPCR), a technology that allows a real-time monitoring of DNA amplification which has proved useful for obtaining a relative measurement, but that only indirectly, through the use of reference standards, provides an absolute quantification.... (Source: European Journal of Histochemistry)
Source: European Journal of Histochemistry - October 18, 2019 Category: Biomedical Science Source Type: research
Hematopoietic stem cells debut in embryonic lymphomyeloid tissues of elasmobranchs
The evolutionary initiation of the appearance in lymphomyeloid tissue of the hemopoietic stem cell in the earliest (most primitive) vertebrate model,i.e. the elasmobranch (chondroichthyan)Torpedo marmorata Risso, has been studied. The three consecutive developmental stages of torpedo embryos were obtained by cesarean section from a total of six pregnant torpedoes. Lymphomyeloid tissue was identified in the Leydig organ and epigonal tissue. The sections were treated with monoclonal anti-CD34 and anti-CD38 antibodies to detect hematopoietic stem cells. At stage I (2-cm-long embryos with external gills) and at stage II (3-4 c...
Source: European Journal of Histochemistry - September 29, 2019 Category: Biomedical Science Source Type: research
Bovine pericardium membrane, gingival stem cells, and ascorbic acid: a novel team in regenerative medicine
Recently, the development and the application of 3D scaffold able to promote stem cell differentiation represented an essential field of interest in regenerative medicine. In particular, functionalized scaffolds improve bone tissue formation and promote bone defects repair. This research aims to evaluate the role of ascorbic acid (AS) supplementation in anin vitro model, in which a novel 3D-scaffold, bovine pericardium collagen membrane called BioRipar (BioR) was functionalized with human Gingival Mesenchymal Stem Cells (hGMSCs). As extensively reported in the literature, AS is an essential antioxidant molecule involved in...
Source: European Journal of Histochemistry - September 25, 2019 Category: Biomedical Science Source Type: research
Expressions of TLR4, MyD88, IRAK4 and NF- κΒ in the oviduct of Chinese brown frog (Rana dybowskii)
In this study, we continued to investigate the localizations and expression levels of cytokines including toll-like receptor 4 (TLR4) and its related factors, myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-associated kinase (IRAK) and nuclear factor kappa B (NF- κΒ) in the oviduct ofRana dybowskiiduring the breeding period and pre-hibernation. Morphological results showed that there were significant differences in oviductal weight and pipe diameter with significantly higher values in pre-hibernation than those of the breeding period. Strong immunostaining of TLR4, MyD88, IRAK4 and NF- κΒ were observe...
Source: European Journal of Histochemistry - September 25, 2019 Category: Biomedical Science Source Type: research
Molecular characterization of nephron progenitors and their early epithelial derivative structures in the nephrogenic zone of the canine fetal kidney
This study was focused to characterize the expression of potential markers of canine NPs and their derivatives by immuno-fluorescence and western blot analysis. Transcription factors (TFs) SIX1 and SIX2, well-characterized human NP markers, were expressed in NPs surrounding the ureteric bud in the CFK. Canine NPs also expressed ITGA8 and NCAM1, surface markers previously used to isolate NPs from the mouse and human fetal kidneys. TF, PAX2 was detected in the ureteric bud, NPs and their derivative structures such as renal vesicle and S-shaped body. This study highlights the similarities in dog, mouse and human renal develop...
Source: European Journal of Histochemistry - September 23, 2019 Category: Biomedical Science Source Type: research
Clathrin mediated endocytosis - Methods and Protocols
Clathrin is one of the interesting “moonlighting proteins” which perform multiple functions relevant to biochemical or biophysical aspects. It can be considered the master regulator of vesicular trafficking being the main player of the Clathrin-Mediated Endocytosis (CME)..... (Source: European Journal of Histochemistry)
Source: European Journal of Histochemistry - September 16, 2019 Category: Biomedical Science Source Type: research
Presence of N-acetylgalactosamine/galactose residues on bronchioloalveolar cells during rat postnatal development
In mammals, the alveolarization process develops predominantly after birth. Airway cells display a complex assemblage of glycans on their surface. These glycans, particularly terminal glycan extensions, are important effective carriers of information that change during the differentiation process. Nevertheless, few systematic data are reported about the cell surface sugar residue content during post-natal lung development. In the present work, we aimed to identify and semi-quantify N-acetylgalactosamine (GalNAc)/galactose (Gal) residues on the bronchioloalveolar cell surface in rat lung sections from 1-, 4-, 8- day old and...
Source: European Journal of Histochemistry - September 10, 2019 Category: Biomedical Science Source Type: research
Ferritin expression in the periodontal tissues of primates
The objective of this study was to determine the expression and distribution of ferritin in the periodontal tissuesof primates.First, we assessed the expression of ferritin in primary cultured cells isolated from human periodontal tissues using the polymerase chain reaction and immunofluorescent staining. Second, we investigated the expression and distribution of ferritin in the periodontal tissues ofMacaca fascicularis, human gingival tissues, and human gingival carcinoma tissues using immunohistochemistry.Both protein and mRNA of ferritin were constitutively present in human primary cultured cells, including those from t...
Source: European Journal of Histochemistry - September 3, 2019 Category: Biomedical Science Source Type: research
Effects of physical training on sarcomere lengths and muscle-tendon interface of the cervical region in an experimental model of menopause
The aim of this study was to describe the structural and ultrastructural aspects of the myotendinous junction (MTJ) and the proximal and distal sarcomeres of the sternomastoid of aged Wistar rats subjected to an experimental model of menopause and swimming training. A total of 20 female elderly rats were divided into the following four groups (n=5 in each group): sedentary/no-menopausal (SNM), trained/no-menopausal (TNM), sedentary/menopausal (SM), and trained/menopausal (TM). The MTJ samples were dissected and analyzed using transmission electron microscopy. We showed that the TNM Group rats exhibited changes in morpholog...
Source: European Journal of Histochemistry - August 6, 2019 Category: Biomedical Science Source Type: research
Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine ™ RNAiMAX and the HepG2-specific, polymer-based GenMute™, in two cellular models of human hepatoma, HepG2 and Huh7.5. In the first part, we assessed transfection efficiency of a fluorescent Cy3-labeled negative control siRNA by cell imaging analysis; we found that cells treated with GenMute pres ent a higher uptake of the fluorescent nega...
Source: European Journal of Histochemistry - August 5, 2019 Category: Biomedical Science Source Type: research
Peribiliary gland damage due to liver transplantation involves peribiliary vascular plexus and vascular endothelial growth factor
In conclusion, PBGs and associated PVP are damaged in transplanted extrahepatic bile ducts; however, an activation of the PBG niche takes place and is characterized by proliferation and VEGF-A expression. This response could have a relevant role in reconstituting biliary epithelium and vascular plexus and could be implicated in the genesis of non-anastomotic strictures. (Source: European Journal of Histochemistry)
Source: European Journal of Histochemistry - May 10, 2019 Category: Biomedical Science Source Type: research
Immunohistochemical identification of resistin in the uterus of ewes subjected to different diets: Preliminary results
Resistin is a polypeptide hormone of the adipokine-family, primarily, but not exclusively, produced by the adipose tissue. Recent studies suggested that resistin may affect the male and female reproductive activity. The study aim was to immunohistochemically evaluate the presence and distribution of resistin in the ovine uterus. Uterine samples were collected from two groups of ewes at the end of an experimental trial during which the animals of the first group (CTRL) were fed only by grazing while those of the second one (EXP) were supplemented with barley and corn. Using a monoclonal antibody against resistin, tested by ...
Source: European Journal of Histochemistry - May 3, 2019 Category: Biomedical Science Source Type: research
miR-132 regulates the expression of synaptic proteins in APP/PS1 transgenic mice through C1q
Cognitive impairment in Alzheimer ’s disease (AD) is usually accompanied by synaptic loss in both the hippocampus and neocortex. In the early stage of AD, amyloid β-induced synapse changes is the main reason, while in the later stage, the accumulation of Tau protein promotes synapse degeneration as the key factor leading to demen tia. MicroRNA (miRNA) is closely related to the expression changes of many AD-related genes. One of the most abundant brain-enriched miRNAs is miR-132, which has been shown to regulate both neuron morphogenesis and plasticity. It has been reported that miR-132 is significantly reduced in the br...
Source: European Journal of Histochemistry - May 2, 2019 Category: Biomedical Science Source Type: research
Embedding cell monolayers to investigate nanoparticle-plasmalemma interactions at transmission electron microscopy
Transmission electron microscopy is the technique of choice to visualize the spatial relationships between nanoconstructs and cells and especially to monitor the uptake process of nanomaterials. It is therefore crucial that the cell surface be preserved in its integrity, to obtain reliable ultrastructural evidence: the plasmalemma represents the biological barrier the nanomaterials have to cross, and the mode of membrane-nanoconstruct interaction is responsible for the intracellular fate of the nanomaterials. In this paper, we describe a simple and inexpensive method to process cell monolayers for ultrastructural morpholog...
Source: European Journal of Histochemistry - March 28, 2019 Category: Biomedical Science Source Type: research
