Background and Aims-The isolation and culture of primary enteric neurons is a difficult process and yields a small number of neurons. We developed fetal (IM-FEN) and postnatal (IM-PEN) enteric neuronal cell lines using the H-2K b -tsA58 transgenic mice that have a temperature sensitive mutation of the SV-40 large tumor antigen gene under the control of an interferon γ-inducible H-2K b promoter element.

The bone morphogenetic protein (BMP) family is a class of transforming growth factor (TGF-␤) superfamily molecules that have been implicated in neuronal differentiation. We studied the effects of BMP2 and glial cell line-derived neurotrophic factor (GDNF) on inducing differentiation of enteric neurons and the signal transduction pathways involved. Studies were performed using a novel murine fetal enteric neuronal cell line (IM-FEN) and primary enteric neurons. Enteric neurons were cultured in the presence of vehicle, GDNF (100 ng/ml), BMP2 (10 ng/ml), or both (GDNF ϩ BMP2), and differentiation was assessed by neurite length, markers of neuronal differentiation (neurofilament medium polypeptide and ␤-III-tubulin), and neurotransmitter expression [neuropeptide Y (NPY), neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase (TH), choline acetyltransferase (ChAT) and Substance P]. BMP2 increased the differentiation of enteric neurons compared with vehicle and GDNF-treated neurons (P Ͻ 0.001). BMP2 increased the expression of the mature neuronal markers (P Ͻ 0.05). BMP2 promoted differentiation of NPY-, nNOS-, and TH-expressing neurons (P Ͻ 0.001) but had no effect on the expression of cholinergic neurons (ChAT, Substance P). Neurons cultured in the presence of BMP2 have higher numbers of TH-expressing neurons after exposure to 1-methyl 4-phenylpyridinium (MPP ϩ ) compared with those cultured with MPP ϩ alone (P Ͻ 0.01). The Smad signal transduction pathway has been implicated in TGF-␤ signaling. BMP2 induced phosphorylation of Smad1, and the effects of BMP2 on differentiation of enteric neurons were significantly reduced in the presence of Smad1 siRNA, implicating the role of Smad1 in BMP2-induced differentiation. The effects of BMP2 on catecholaminergic neurons may have therapeutic implications in gastrointestinal motility disturbances.immorto; glial cell line-derived neurotrophic factor; bone morphogenic protein 2; Smad1 ENTERIC NEURONS ARE DERIVED from vagal, sacral, and upper thoracic neural crest cells that migrate into the gut, where they proliferate and then differentiate into a variety of distinct cell types (2,17,22,23). Several growth factors influence the proliferation, differentiation, and survival of the enteric nervous system (ENS), including glial cell line-derived neurotrophic factor (GDNF), neurturin, neurotrophin-3, and Sonic hedgehog (14, 18). Bone morphogenetic proteins (BMPs), members of the transforming growth factor-␤ (TGF-␤) superfamily, are regulatory molecules that play critical roles in the formation and regionalization of the gut before its colonization by crest-derived cells (26,27). BMP2 and 4 continue to be expressed in the fetal gut and regulate the specification, migration, differentiation, and aggregation of enteric neurons (12,13,15,24).We have recently developed an enteric neuronal cell line that has characteristics similar to primary enteric neurons (4). We sought to determine the types of growth factors that would result in differentiation of this novel enteric neuronal cel...

Background Development of pre-transplantation islet culture strategies that preserve or enhance β-cell viability would eliminate the requirement for the large numbers of islets needed to restore insulin independence in type 1 diabetes patients. We investigated whether glial cell line-derived neurotrophic factor (GDNF) could improve human islet survival and post-transplantation function in diabetic mice. Methods Human islets were cultured in medium supplemented with or without GDNF (100 ng/ml) and in vitro islet survival and function assessed by analyzing β-cell apoptosis and glucose stimulated insulin release. In vivo effects of GDNF were assessed in streptozotocin-induced diabetic nude mice transplanted under the kidney capsule with 2000 islet equivalents of human islets pre-cultured in medium supplemented with or without GDNF. Results In vitro, human islets cultured for 2–10 days in medium supplemented with GDNF showed lower β-cell death, increased Akt phosphorylation and higher glucose-induced insulin secretion than islets cultured in vehicle. Human islets pre-cultured in medium supplemented with GDNF restored more diabetic mice to normoglycemia and for a longer period after transplantation than islets cultured in vehicle. Conclusions Our study shows that GDNF has beneficial effects on human islet survival and could be used to improve islet post-transplantation survival.

Data is a crucial resource for every business, and it must be protected both during storage and transmission. One efficient way of securing data and transferring it is through digital watermarking, where data is hidden inside a medium like text, audio, or video. Video watermarking is visible or invisible embedded data on a video in a logo, text, or video copyright disclaimer. In this proposed paper, the goal is to analyze the characteristics of video watermarking algorithms and the different metrics used for them. It deals with the extent to which the different requirements can be fulfilled, taking into consideration the conflicts between them and the practical challenges of video watermarking in terms of attacks like geometric attacks and non-geometric attacks. It also focuses on the process of watermarking a video. Recent advances in data security indicate that employing a video watermarking technology to transmit private data will be an effective method of transmitting sensitive data.

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