In previous studies we have shown that protein kinase inhibitors and extracellular calcium can affect dramatically the assembly of tight junctions (TJ) and the localization of the TJ protein cingulin at sites of cell-cell contact in renal epithelial (MDCK) cells. To characterize in more detail the relationships between kinase activity and junction organization, we have studied the effects of the protein kinase C agonist phorbol myristate acetate (PMA) on the intracellular localization of cingulin, E-cadherin, desmoplakin and actin microfilaments in confluent MDCK monolayers. To study cingulin phosphorylation, MDCK cells were metabolically labelled with [32P]orthophosphate and immunoprecipitates were prepared with anti-cingulin antiserum. We show here that cingulin is phosphorylated in vivo on serine, and its specific phosphorylation is not significantly changed by treatment of confluent MDCK monolayers with PMA, with the protein kinase inhibitor H-7, or with the calcium chelator EGTA. Metabolic ...

Park NX-Bio enables that with its innovative in-liquid imaging Scanning Ion Conductance Microscopy (SICM) and its highly acclaimed Atomic Force Microscopy (AFM) technology.

in Anticancer Research (2007), 27. The viability, cytolysis and apoptosis-mediated cellular death induced by giganteosides D and E (Gig-D and Gig-E) and hederacolchisides A and A1 (Hcol-A and Hcol- A1) were analysed in HL-60 cells ... [more ▼]. The viability, cytolysis and apoptosis-mediated cellular death induced by giganteosides D and E (Gig-D and Gig-E) and hederacolchisides A and A1 (Hcol-A and Hcol- A1) were analysed in HL-60 cells. Materials and Methods: the end-point metabolic (WST1) and lactate dehydrogenase (LDH) assays were used. Cell cycle analysis and apoptosis were measured by flow cytometry, DNA laddering and caspase 3 analyses. Results: the HL-60 cell line was more sensitive to Hcol-A1 and Gig-D (IC50 3-5 ÌM) than to Gig-E and Hcol-A (IC50 8-13 ÌM; WST1 assay). This was related to LDH release. The induction of apoptosis could be detected without caspase 3 activation after 24 h of treatment. DNA fragmentation could be detected only with Gig-D. With Hcol- A1 and Gig-D, an ...

Supplementary Materials Online Supplement supp_45_3_534__index. noticed an apoptosis-mediated and necrosis-mediated depletion ( 90%) from the receiver Compact disc103poperating-system DC subset, in support of a 50C60% depletion of receiver Compact disc11bpos DCs from lung parenchymal tissues on Times 3 and 5, whereas receiver lung and alveolar macrophages had been significantly less radiosensitive, showing an around 50% depletion by Times 14C21 after treatment. A repopulation of lung tissues with donor DC subsets acquired occurred by Times 10 and 28 for Compact disc11bpos DCs and Compact disc103poperating-system DCs, whereas lung and alveolar macrophages were repopulated by 6 and 10 weeks after treatment. Furthermore, chlamydia of mice with further accelerated the turnover of lung lung and DCs macrophage subsets. Our data illustrate the vulnerability of lung Compact disc103poperating-system Compact disc11bpos and DCs DCs to irradiation, and indicate an accelerated turnover of lung DC subsets ...

TY - JOUR. T1 - Micropipet-Based Navigation in a Microvascular Model for Imaging Endothelial Cell Topography Using Scanning Ion Conductance Microscopy. AU - Taira, Noriko. AU - Nashimoto, Yuji. AU - Ino, Kosuke. AU - Ida, Hiroki. AU - Imaizumi, Takuto. AU - Kumatani, Akichika. AU - Takahashi, Yasufumi. AU - Shiku, Hitoshi. N1 - Publisher Copyright: © PY - 2021. Y1 - 2021. N2 - Scanning ion conductance microscopy (SICM) has enabled cell surface topography at a high resolution with low invasiveness. However, SICM has not been applied to the observation of cell surfaces in hydrogels, which can serve as scaffolds for three-dimensional cell culture. In this study, we applied SICM for imaging a cell surface in a microvascular lumen reconstructed in a hydrogel. To achieve this goal, we developed a micropipet navigation technique using ionic current to detect the position of a microvascular lumen. Combining this navigation technique with SICM, endothelial cells in a microvascular model and blebs were ...

Scanning electrochemical microscopy (SECM) is a powerful technique for examining the diffusive, convective, and migratory transport of solutes. In SECM, an ultramicroelectrode (UME), attached to piezoelectric positioners, is mobile in three dimensions. The UME can be positioned close to an interface with submicron precision, and can probe the topography, reactivity, or permeability of that interface with high spatial resolution (Bard et al., 1991b; Barker et al., 1999). SECM has been applied to the study of a number of synthetic membranes and biomaterials including skin (Bath et al., 1998; Scott et al., 1991; 1993a, b; 1995), dentine (Macpherson et al., 1995a, b; Unwin et al., 1997), and bilayer lipid membranes (Matsue et al., 1994). SECM has the advantage over scanning ion conductance microscopy, which has found some application in the investigation of membrane transport (Hansma et al., 1989; Korchev et al., 1997), in that it can selectively detect both neutral and charged species, rather than ...

Scanning electrochemical microscopy (SECM) is a powerful technique for examining the diffusive, convective, and migratory transport of solutes. In SECM, an ultramicroelectrode (UME), attached to piezoelectric positioners, is mobile in three dimensions. The UME can be positioned close to an interface with submicron precision, and can probe the topography, reactivity, or permeability of that interface with high spatial resolution (Bard et al., 1991b; Barker et al., 1999). SECM has been applied to the study of a number of synthetic membranes and biomaterials including skin (Bath et al., 1998; Scott et al., 1991; 1993a, b; 1995), dentine (Macpherson et al., 1995a, b; Unwin et al., 1997), and bilayer lipid membranes (Matsue et al., 1994). SECM has the advantage over scanning ion conductance microscopy, which has found some application in the investigation of membrane transport (Hansma et al., 1989; Korchev et al., 1997), in that it can selectively detect both neutral and charged species, rather than ...

Application archive. Nov. 26, 2018Applications Scanning Ion Conductance Microscopy - Scanning Electrochemical Microscopy Since the inception of scanning tunneling microscopy (STM) [1],

Podcast: Play in new window , Download. Subscribe: Android , RSS. In this weeks episodes, I discuss the presentation and management of acute, chronic and allergic fungal rhinosinusitis.. ...

The avian basilar papilla is composed of hair and supporting cells arranged in a regular pattern in which the hair cells are surrounded and isolated from each other by supporting cell processes. This arrangement of cells, in which the apical borders of hair cells do not contact one another, may be generated by contact-mediated lateral inhibition. Little is known, however, about the way in which hair and supporting cells are organized during development. Whole mounts double-labeled with antibodies to the 275 kDa hair-cell antigen and the tight junction protein cingulin were therefore used to examine the development of cell patterns in the basilar papilla. Hair cells that contact each other at their apical borders are seen during early development, especially on embryonic days (E) 8 and 9, but are no longer observed after E12. Hair and supporting cell patterns were analyzed in three different areas of the papilla at E9 and E12. In two of these regions between E9 and E12, the ratio of supporting ...

Allergic fungal rhinosinusitis (AFRS) is a unique variety of chronic polypoid rhinosinusitis usually in atopic individuals, characterized by presence of eosinophilic mucin and fungal hyphae in paranasal sinuses without invasion into surrounding mucosa. It has emerged as an important disease involving a large population across the world with geographic variation in incidence and epidemiology. The disease is surrounded by controversies regarding its definition and etiopathogenesis. A working group on

Upon viral infection, the major defense mounted by the host innate immune system is activation of the IFN- and apoptosis-mediated antiviral pathway. In order to complete their life cycle, viruses that are obligatory intracellular parasites must modulate these host immune responses. We have previously shown that the γHV68 latency-associated M2 protein effectively downregulates STAT1 and STAT2, resulting in the inhibition of type I and II IFN-mediated transcriptional activation. Here, we demonstrate that M2 interacts with ATM, a DNA damage signal transducer, and the DDB1/COP9/cullin DNA damage effector complex. This interaction blocked DNA damage-sensing activity as well as DNA damage repair activity, thereby rendering cells resistant to DNA damage-induced apoptosis. These results indicate that γHV68 encodes M2, a latency-associated gene, to antagonize both IFN- and apoptosis-mediated host innate immunities and thus is important in establishing and maintaining viral latency in infected ...

Our data reveal that the H,K-ATPase ion pump is required for apoptosis-mediated head size and organ scaling throughout the animal. Morphallactic defects in Smed-H,K-ATPase(RNAi) regenerates were observed in: head and pharyngeal resizing; pharyngeal A/P placement; anterior neural, brain and intestinal tract remodeling; and the re-establishment of anterior identity in previously more posterior tissues. All these remodeling defects occurred without inhibiting new tissue proliferation associated with blastema growth (epimorphosis). Therefore, when H,K-ATPase is lost during regeneration, head is made from new tissues and stays small because remodeling failures prevent old tissues from contributing to it, whereas the pharynx stays large because apoptosis fails to sculpt it to the correct size. This suggests that morphallactic remodeling of pre-existing tissues is an independent pathway from epimorphic blastema growth, a distinction that was previously unclear (Agata et al., 2007).. The working ...

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Food allergies, celiac disease, inflammatory bowel disease (IBD), diarrhea and other gastrointestinal diseases have something in common: all have been linked to epithelial barrier loss. The gut epithelial barrier-that critical lining of cells in the gut that must allow nutrients into the body while keeping food-borne microbes out-can be compromised during intestinal inflammation and cause disease. While many of the molecular mechanisms that trigger gastrointestinal diseases remain a mystery, previous research has found that one enzyme, known as myosin light chain kinase (MLCK), plays a critical role. However, MLCK is also essential for critical functions in gut epithelia and other cell types. This makes direct inhibition of MLCK impossible, as it would result in many toxic and systemic side effects. A team led by investigators from Brigham and Womens Hospital has now developed an alternative approach. In a study published in Nature Medicine, the researchers report new evidence suggesting that ...

Shop a large selection of Proteins A-Z products and learn more about Novus Biologicals Claudin-2 Recombinant Protein Antigen Quantity: 100µL:Life Quantity: 100µL.

Claudin-4 (Clostridium perfringens enterotoxin receptor) is a tight junction protein encoded by the gene CLDN4. Expression of Claudin-4 has been associated with either poor prognosis or a more favorable diagnosis, depending on the type of cancer. Claudin-4 has been shown to distinguish adenocarcinoma from malignant mesothelioma with 99% specificity in malignant effusions (1). Claudin-4 overexpression was able to independently predict survival in a breast cancer multivariate analysis as it was associated with poor prognosis, high tumor grade and Her2 expression and was inversely correlated with estrogen receptor staining (2). In luminal breast cancer, the increase of Claudin-4 protein was correlated with the increase of tumor grade and with Ki-67, and thus demonstrated an overall shorter life survival (3). Basal-like tumors also demonstrated overexpression of Claudin-4 (4). Counter to the above breast cancer subtypes, the presence of Claudin-4 in triple negative breast cancer was a biomarker that

The expression of claudin-11 in benign and malignant bladder tissue and the effect of forced expression of claudin-11 on tight junction function and invasiveness of bladder cancer cells were studied. Claudin-11 expression was tested in bladder cancer cell lines (T24/83, RT 112/84 and EJ138) using reverse transcription-polymerase chain reaction (RT-PCR) and in benign and malignant bladder tissue by quantitative RT-PCR and immunohistochemistry. T24/83 cells were transfected with the pcDNA.1/NT-GFP-TOPO vector containing full-length human claudin-11 sequence. Stable-transfected cells overexpressing claudin-11 (T24Cl-11Ex), wild-type cells (T24WT) and the empty plasmid control clone (T24GFP) were compared using transurothelial resistance (TUR), in vitro adhesion, invasion and growth assays. Claudin-11 was strongly expressed in the non-invasive RT112/84 cell line compared to the invasive T24/83 and EJ138 TCC cell lines. Benign bladder tissue demonstrated equal expression of claudin-11 mRNA as ...

Objective Helicobacter pylori strains that express the oncoprotein CagA augment risk for gastric cancer. However, the precise mechanisms through which cag+ strains heighten cancer risk have not been fully delineated and model systems that recapitulate the gastric niche are critical for understanding pathogenesis. Gastroids are three-dimensional organ-like structures that provide unique opportunities to study host-H. pylori interactions in a preclinical model. We used gastroids to inform and direct in vitro studies to define mechanisms through which H. pylori modulates expression of the cancer-associated tight junction protein claudin-7.. ...

Read Claudin-8 Expression in Renal Epithelial Cells Augments the Paracellular Barrier by Replacing Endogenous Claudin-2, The Journal of Membrane Biology on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.

This work investigates the physicochemical properties and in vitro accuracy of a genetically engineered drug delivery system based on elastin-like block recombinamers. The DNA recombinant technics allowed us to create this smart complex polymer containing bioactive sequences for internalization, lysosome activation under acidic pH and blockage of cellular growth by a small peptide inhibitor. The recombinant polymer reversibly self-assembled, when temperature was increased above 15°C, into nanoparticles with a diameter of 72 nm and negative surface charge. Furthermore, smart nanoparticles were showed to enter in the cells via clathrin-dependent endocytosis, and properly blocked phosphorylation and consequent activation of Akt kinase. This system provoked apoptosis-mediated cell death in breast and colorectal cancer cells, which possess higher expression levels of Akt, whereas non-cancerous cells, such as endothelial cells, fibroblasts and mesenchymal stem cells, were not affected. Hence, we ...

Shop a large selection of products and learn more about Claudin-9 Rabbit anti-Human, Polyclonal, Novus Biologicals 100µL; 100µL; Unlabeled.

Claudin-3 is a major protein of tight junctions (TJs) in the intestinal epithelium and is critical for maintaining cell-cell adhesion, barrier function, and epithelium polarity. Recent studies have shown high claudin-3 levels in several solid tumors, but the regulation mechanism of claudin-3 expression remains poorly understood. In the present study, colorectal cancer (CRC) tissues, HT-29 and DLD-1 CRC cell lines, CRC murine model (C57BL/6 mice) and c-kit loss-of-function mutant mice were used. We demonstrated that elevated claudin-3 levels were positively correlated with highly expressed c-kit in CRC tissues based upon analysis of protein expression. In vitro, claudin-3 expression was clearly increased in CRC cells by overexpressed c-kit or stimulated by exogenous recombinant human stem cell factor (rhSCF), while significantly decreased by the treatment with c-kit or c-Jun N-terminal kinase (JNK) inhibitors. Chromatin immunoprecipitation (ChIP) and luciferase reporter assay showed that SCF/c-kit

DESCRIPTION (provided by applicant): Disruption of the cell-cell junction with concomitant changes in the expression of junctional proteins is a hallmark of cancer metastasis and invasion. Role of adherent junction proteins have been studied extensively in cancer, however the role of tight junction proteins is less understood. Claudins are the recently identified tetraspanins, which are integral to the structure and function of tight junctions (TJs). Recent studies have shown changes in expression/cellular localization for claudins during tumorigenesis, however a cause and effect relationship has not been established. Here, we report a highly increased expression for claudin-1 in human primary colon carcinoma and metastatic tissues and cell lines derived from similar sources with relatively frequent nuclear localization. Furthermore, using genetic manipulations of claudin-1 expression in colon cancer cell lines, we demonstrate a role for claudin-1 in the regulation of epithelial to mesenchymal ...