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In vivo whole cell

Nailah Seale, Suvasini Ramaswamy, Yu-Ru Shih, Inder Verma, Shyni Varghese
BACKGROUND: Given the shortage of available organs for whole or partial liver transplantation, hepatocyte cell transplantation has long been considered a potential strategy to treat patients suffering from various liver diseases. Some of the earliest approaches that attempted to deliver hepatocytes via portal vein or spleen achieved little success due to poor engraftment. More recent efforts include transplantation of cell sheets or thin hepatocyte laden synthetic hydrogels. However, these implants must remain sufficiently thin to ensure that nutrients can diffuse into the implant...
June 18, 2018: Transplantation
Xinmei Luo, Sujuan Ye, Qianqian Jiang, Yi Gong, Yue Yuan, Xueting Hu, Xiaolan Su, Wen Zhu
Wnt inhibitory factor‑1 (WIF‑1) is an important antagonist of Wnt/β‑catenin signaling by binding to Wnt ligands. The downregulation of WIF‑1 leads to the development of non‑small cell lung cancer (NSCLC). The upregulation of WIF‑1 significantly inhibits proliferation and induces apoptosis by inhibiting Wnt/β‑catenin signaling in NSCLC. However, the mechanisms underlying the inhibition of Wnt/β‑catenin signaling by WIF‑1‑mediated autophagy are poorly understood. Thus, in this study, we aimed to shed some light into these mechanisms...
June 15, 2018: International Journal of Oncology
Evelyne Gineyts, Nicolas Bonnet, Cindy Bertholon, Marjorie Millet, Aurélie Pagnon-Minot, Olivier Borel, Sandra Geraci, Edith Bonnelye, Martine Croset, Ali Suhail, Cristina Truica, Nicholas Lamparella, Kim Leitzel, Daniel Hartmann, Roland Chapurlat, Allan Lipton, Patrick Garnero, Serge Ferrari, Philippe Clézardin, Jean-Charles Rousseau
Periostin is an extracellular matrix protein that actively contributes to tumor progression and metastasis. Here, we hypothesized that it could be a marker of bone metastasis formation. To address this question, we used two polyclonal antibodies directed against the whole molecule or its C-terminal domain to explore the expression of intact and truncated forms of periostin in the serum and tissues (lung, heart, bone) of wild-type and periostin-deficient mice. In normal bones, periostin was expressed in the periosteum and specific periostin proteolytic fragments were found in bones, but not in soft tissues...
June 18, 2018: Calcified Tissue International
Areumnuri Kim, Sehwan Shim, Min-Jung Kim, Jae Kyung Myung, Sunhoo Park
Radiation exposure severely damages the hematopoietic system. Although several radio-protectors have been proposed to prevent radiation-induced damage, most agents have limited efficacy. In the present study, we investigated whether mesenchymal stem cells (MSCs) could contribute to the expansion of hematopoietic cells and mitigate radiation-induced hematopoietic injury in vitro and in vivo. We found that co-culture with MSCs promoted hematopoietic progenitor/stem cell (HPSCs) maintenance by providing a bone marrow-like microenvironment...
June 18, 2018: Scientific Reports
Susan J Burke, Heidi M Batdorf, David H Burk, Thomas M Martin, Tamra Mendoza, Krisztian Stadler, Wateen Alami, Michael D Karlstad, Matthew J Robson, Randy D Blakely, Randall L Mynatt, J Jason Collier
OBJECTIVE: Pancreatic tissue, and islets in particular, are enriched in expression of the interleukin-1 receptor type I (IL-1R). Because of this enrichment, islet β-cells are exquisitely sensitive to the IL-1R ligands IL-1α and IL-1β, suggesting that signaling through this pathway regulates health and function of islet β-cells. METHODS: Herein, we report a targeted deletion of IL-1R in pancreatic tissue (IL-1RPdx1-/- ) in C57BL/6J mice and in db/db mice on the C57 genetic background...
June 6, 2018: Molecular Metabolism
Xiaoye Wang, Genglin Guo, Xin Zhang, Ming Li, Kai Xiao, Chuanhuo Hu, Xun Li
RFamide-related peptide-3 (RFRP-3) and its receptor (GPR147) play an important role in reproduction regulation in mammals. To understand the role of RFRP-3 in male reproductive function of epididymis, we first investigated the expression changes in RFRP-3 and its receptor at different stages of development, that is, postnatal day 20 (P20), 40 (P40), 60 (P60) and 80 (P80). Our results showed that fluctuations in the expression of GnIH and GPR147 during postnatal development occurred, and the highest epididymal GnIH and GPR147 expression were both detected in P60...
June 5, 2018: Theriogenology
Taylor Pini, Tamara Leahy, Simon P de Graaf
Semen cryopreservation is an important tool for artificial breeding, species conservation and human reproductive medicine. However, sublethal freezing damage remains an important limitation of the cryopreservation process, inevitably leading to reduced fertility in vivo. This review explores several facets of sublethal freezing damage, touching on cryocapacitation, the generation of reactive oxygen species and alterations to sperm proteins, lipids and sugars. The effects of sublethal freezing damage on sperm performance in vivo are also discussed, examining fertile lifespan and interaction with female reproductive tract immune cells, mucus and oviductal cells...
June 11, 2018: Theriogenology
Natascha Schaefer, Fang Zheng, Johannes van Brederode, Alexandra Berger, Sophie Leacock, Hiromi Hirata, Christopher J Paige, Robert J Harvey, Christian Alzheimer, Carmen Villmann
Mutations in GlyR α1 or β subunit genes in humans and rodents lead to severe startle disease characterized by rigidity, massive stiffness and excessive startle responses upon unexpected tactile or acoustic stimuli. The recently characterized startle disease mouse mutant shaky carries a missense mutation (Q177K) in the β8-β9 loop within the large extracellular N-terminal domain of the GlyR α1 subunit. This results in a disrupted hydrogen bond network around K177 and faster GlyR decay times. Symptoms in mice start at postnatal day 14 and increase until premature death of homozygous shaky mice around 4-6 weeks after birth...
2018: Frontiers in Molecular Neuroscience
Amy E O'Connell, Fanny Zhou, Manasvi S Shah, Quinn Murphy, Hannah Rickner, Judith Kelsen, John Boyle, Jefferson J Doyle, Bharti Gangwani, Jay R Thiagarajah, Daniel S Kamin, Jeffrey D Goldsmith, Camilla Richmond, David T Breault, Pankaj B Agrawal
Homozygous nonsense mutations in WNT2B were identified in three individuals from two unrelated families with severe, neonatal-onset osmotic diarrhea after whole-exome sequencing was performed on trios from the two families. Intestinal biopsy samples from affected individuals were used for histology and immunofluorescence and to generate enteroids ex vivo. Histopathologic evaluation demonstrated chronic inflammatory changes in the stomach, duodenum, and colon. Immunofluorescence demonstrated diminished staining for OLFM4, a marker for intestinal stem cells (ISCs)...
June 4, 2018: American Journal of Human Genetics
An Zeng, Hua Li, Longhua Guo, Xin Gao, Sean McKinney, Yongfu Wang, Zulin Yu, Jungeun Park, Craig Semerad, Eric Ross, Li-Chun Cheng, Erin Davies, Kai Lei, Wei Wang, Anoja Perera, Kate Hall, Allison Peak, Andrew Box, Alejandro Sánchez Alvarado
Proliferating cells known as neoblasts include pluripotent stem cells (PSCs) that sustain tissue homeostasis and regeneration of lost body parts in planarians. However, the lack of markers to prospectively identify and isolate these adult PSCs has significantly hampered their characterization. We used single-cell RNA sequencing (scRNA-seq) and single-cell transplantation to address this long-standing issue. Large-scale scRNA-seq of sorted neoblasts unveiled a novel subtype of neoblast (Nb2) characterized by high levels of PIWI-1 mRNA and protein and marked by a conserved cell-surface protein-coding gene, tetraspanin 1 (tspan-1)...
June 14, 2018: Cell
Tong Shao, Wei Shi, Jia-Yu Zheng, Xiao-Xiao Xu, Ai-Fu Lin, Li-Xin Xiang, Jian-Zhong Shao
CD58 and CD2 have long been known as a pair of reciprocal adhesion molecules involved in the immune modulations of CD8+ T and NK-mediated cellular immunity in humans and several other mammals. However, the functional roles of CD58 and CD2 in CD4+ T-mediated adaptive humoral immunity remain poorly defined. Moreover, the current functional observations of CD58 and CD2 were mainly acquired from in vitro assays, and in vivo investigation is greatly limited due to the absence of a Cd58 homology in murine models...
2018: Frontiers in Immunology
Konstantin Stoletov, Lian Willetts, Robert J Paproski, David J Bond, Srijan Raha, Juan Jovel, Benjamin Adam, Amy E Robertson, Francis Wong, Emma Woolner, Deborah L Sosnowski, Tarek A Bismar, Gane Ka-Shu Wong, Andries Zijlstra, John D Lewis
Metastasis is the most lethal aspect of cancer, yet current therapeutic strategies do not target its key rate-limiting steps. We have previously shown that the entry of cancer cells into the blood stream, or intravasation, is highly dependent upon in vivo cancer cell motility, making it an attractive therapeutic target. To systemically identify genes required for tumor cell motility in an in vivo tumor microenvironment, we established a novel quantitative in vivo screening platform based on intravital imaging of human cancer metastasis in ex ovo avian embryos...
June 14, 2018: Nature Communications
Sirikamon Koosakulnirand, Phornpun Phokrai, Kemajittra Jenjaroen, Rosemary A Roberts, Pongsak Utaisincharoen, Susanna J Dunachie, Paul J Brett, Mary N Burtnick, Narisara Chantratita
Burkholderia pseudomallei is a flagellated Gram-negative bacterium which is the causative agent of melioidosis. The disease poses a major public health problem in tropical regions and diabetes is a major risk factor. The high mortality rate of melioidosis is associated with severe sepsis which involves the overwhelming production of pro-inflammatory cytokines. Bacterial flagellar protein (flagellin) activates Toll-like receptor 5 (TLR5)-mediated innate immune signaling pathways and induces adaptive immune response...
2018: PloS One
Mi Ae Kang, Wonwoo Kim, Hye-Ram Jo, Young-Joo Shin, Moon-Hong Kim, Jae-Hoon Jeong
Cyclin-dependent kinases (CDK) are considered to be potential targets of anticancer drugs that can interrupt the uncontrolled division of cancer cells. In this study, we selected two selective CDK inhibitors, AT7519 and SNS‑032, from current clinical trials and examined their anticancer and radiosensitizing effects in a cervical cancer model. SNS‑032 was found to be more potent than AT7519, with a lower half maximal inhibitory concentration (IC50) value. Both AT7519 and SNS‑032 induced the apoptosis, premature senescence and cytostasis of cervical cancer cells, which led to the attenuation of tumor growth in vivo...
May 31, 2018: International Journal of Oncology
Chie Ishikawa, Masachika Senba, Naoki Mori
Adult T‑cell leukemia/lymphoma (ATLL) is a disorder involving human T-cell leukemia virus type 1 (HTLV‑1)-infected T‑cells characterized by increased clonal neoplastic proliferation. PDZ-binding kinase (PBK) [also known as T‑lymphokine-activated killer cell-originated protein kinase (TOPK)] is a serine/threonine kinase expressed in proliferative cells and is phosphorylated during mitosis. In this study, the expression and phosphorylation of PBK/TOPK were examined by western blot analysis and RT‑PCR...
June 1, 2018: International Journal of Oncology
Hussam Hayani, Inseon Song, Alexander Dityatev
The neural extracellular matrix (ECM) is enriched with hyaluronic acid, chondroitin sulfate proteoglycans (CSPGs) and the glycoprotein tenascin-R, which play important roles in synaptic plasticity, as shown by studies of the CA1 region of the hippocampus. However, ECM molecules are strongly expressed in the CA2 region, which harbors a high number of fast-spiking interneurons (FSIs) surrounded by a particularly condensed form of ECM, perineuronal nets. Despite this intriguing peculiarity, the functional role of ECM in the CA2 region is mostly unknown...
2018: Frontiers in Cellular Neuroscience
Daniel B Yaeger, Emma J Coddington
Neurons in the medullary reticular formation are involved in the control of postural and locomotor behaviors in all vertebrates. Reticulospinal neurons in this brain region provide one of the major descending projections to the spinal cord. Although neurons in the newt medullary reticular formation have been extensively studied using in vivo extracellular recordings, little is known of their intrinsic biophysical properties or of the underlying circuitry of this region. Using whole-cell patch-clamp recordings in brain slices containing the rostromedial reticular formation (rmRF) from adult male newts, we observed spontaneous miniature outward currents (SMOCs) in approximately two-thirds of neurons...
June 13, 2018: Journal of Neurophysiology
Benjamin Piña, Laia Navarro, Carlos Barata, Demetrio Raldúa, Rubén Martínez, Marta Casado
The genome revolution represents a complete change on our view of biological systems. The quantitative determination of changes in all major molecular components of the living cells, the "omics" approach, opened whole new fields for all health sciences. Genomics, transcriptomics, proteomics, metabolomics, and others, together with appropriate prediction and modeling tools, will mark the future of developmental toxicity assessment both for wildlife and humans. This is especially true for disciplines, like teratology, which rely on studies in model organisms, as studies at lower levels of organization are difficult to implement...
2018: Methods in Molecular Biology
Kyung-Ran Park, Eun-Cheol Kim, Jin Tae Hong, Hyung-Mun Yun
Rationale: Characterizing the regulation of bone-resorbing osteoclasts is central to the understanding of the pathogenesis and treatment of bone diseases, such as osteoporosis and periodontitis. 5-hydroxytryptamine (5-HT) has drawn considerable attention for its role in bone; however, it remains unknown whether the intracellular signaling of 5-HT receptors (5-HTRs) is linked to any of the regulatory mechanisms in osteoclasts. Herein, we report 5-HT6 R to be a key regulatory receptor for osteoclastogenesis. Methods: In order to explore the critical role of 5-HT6 R in bone-resorbing osteoclasts, in vitro experiments were performed using mouse whole bone marrow cells isolated from femora and tibiae and In vivo animal experiments were performed using 5-HT6 R-deficient (5-HT6 RKO-/- ) mice, bone resorption mice model, and osteoporosis mice model...
2018: Theranostics
Naoki Komatsu, Kenta Terai, Ayako Imanishi, Yuji Kamioka, Kenta Sumiyama, Takashi Jin, Yasushi Okada, Takeharu Nagai, Michiyuki Matsuda
Genetically encoded biosensors based on the principle of Förster resonance energy transfer comprise two major classes: biosensors based on fluorescence resonance energy transfer (FRET) and those based on bioluminescence energy transfer (BRET). The FRET biosensors visualize signaling-molecule activity in cells or tissues with high resolution. Meanwhile, due to the low background signal, the BRET biosensors are primarily used in drug screening. Here, we report a protocol to transform intramolecular FRET biosensors to BRET-FRET hybrid biosensors called hyBRET biosensors...
June 12, 2018: Scientific Reports
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