A series of stiripentol (SIP) analogues namely, 2-1(1E)-1-(1,3-benzodioxol-5-yl)-4,4-dimethylpent-1-en-3-ylidene]-N-(aryl/H)hydrazinecarboxamides 7a-h, (+/-)-(5RS)-N-(aryl/H)-(1,3-benzodioxol-5-yl)-3-tert-butyl-4,5-dihydro-1H-pyrazole-1-carboxamides (+/-)-8a-h, and (+/-)-[(5RS)-(1,3-benzodioxol-5-yl)-3-tert-butyl-4,5-dihydro-1H-pyrazol-1-yl](aryl)methanones (+/-)-13a-f was synthesized by adopting appropriate synthetic routes and was pharmacologically evaluated in the preliminary anticonvulsant screens. The selected bioactive new chemical entities were subjected to ED50 determination and neurotoxicity evaluation. The most active congeners are 7h in MES screen and (+/-)-13b in scPTZ screen which displayed ED50 values of 87 and 110 mg/kg, respectively, as compared to that of STP (ED50 = 277.7 and 115 mg/kg in MES and scPTZ, respectively). (C) 2011 Elsevier Masson SAS. All rights reserved.
Signaling molecules in the gastrointestinal (GI) tract, as released from intrinsic, or extrinsic neurons, or from local endocrine cells may serve as positive or negative growth factors, and it has been suggested that such could participate also in colorectal carcinogenesis/cancer progression. Sporadic colorectal cancer arises from an initially benign adenoma, which, in turn, develops from the stem cell compartment, located in the bottom of the crypts of the colorectal mucosa. It was recently demonstrated in rat that intrinsic denervation of the colon appeared to be protective against chemically induced carcinogenesis. Of the various GI signaling molecules, noradrenaline (NA) and substance P (SP) may be of particular importance as growth factors involved in colorectal cancer. In the current issue of Neurogastroenterology and Motility, Graf et al. demonstrate that in benign, human colon polyps, there was a loss of innervation compared with adjacent mucosa, affecting efferent, noradrenergic, as well as sensory, SP-ergic fibers, while there was an increase in SP-immunoreactive non-neuronal cells in the polyps. The results obtained could suggest that loss of mucosal innervation, due to e.g. luminal, pro-inflammatory stimuli, could result in unbalanced pro-tumorigenic stimulation of the stem cell region by non-neuronal SP. The current findings may be important for the further understanding of the development of sporadic colorectal cancer.
Various markers of the cholinergic system (like e.g. choline acetyltransferase) were demonstrated by immunohistochemistry in, seemingly, beta-cells of rat pancreas. The findings may suggest an autocrine role of acetylcholine for the beta-cells. (C) 2011 Elsevier B.V. All rights reserved.
Background
Tumor associated macrophages (TAMs) are often present at a high level in colorectal cancer (CRC) but whether the presence of TAMs in CRC is good or bad is unclear. Macrophages can be categorized into two main subgroups, M1 or M2, that display pro – and anti-inflammatory properties respectively. An improved knowledge of the different macrophage phenotypes will broaden the understanding of their involvement in CRC. We have used an in vitro model to study the effects of human M1 and M2 macrophages on the growth and cell cycle of colon cancer cell lines.
Method
Human monocytes were differentiated into M1 or M2 macrophages and conditioned media was collected. Effects of the conditioned media were measured on the colon cancer cell lines HT-29 and CACO-2 in regards to proliferation, cell cycle distribution and apoptosis. A protein array was used to analyze the released amount of 42 different cytokines from M1 and M2 macrophages into the collected conditioned media.
Results
Growth arrest was induced in HT-29 and CACO-2 by M1 conditioned media, while M2 conditioned media had no major effect. Analysis of cell cycle distribution and apoptosis in HT-29 cells revealed that treatment with M1 conditioned media on these cells increased apoptosis and caused a disturbed cell cycle with accumulation in G0/G1 and G2/M and a corresponding reduction in S-phase. The protein array revealed several cytokines expressed in M1 with potential inhibitory growth effects. We treated HT-29 cells with two of the candidates, TNF-a and CXCL9, but neither induced growth arrest.
Conclusion
M1 – but not M2-macrophages had a major inhibitory effect on the growth of the colon cancer cell lines HT-29 and CACO-2 and suggest a role for M1 macrophages in anti-tumor activity and possible favorable outcome for CRC patients.
At immunolocalization of experimental tumors, idiotypic monoclonal antibodies, such as TS1 against cytokeratin 8, can be used to carry and deposit in vivo terapeutics in the tumor. These carriers also remain in the circulation and may cause negative side-effects in other tissues.In this report, several derivatives of the antiidiotypic antibody anti-TS1 were produced and tested for their clearing capacity of the idiotypic carrier antibody TS1. Intact monoclonal anri-TS1, scFv of a anti-TS1 and anti-TS1 Fab and Fab'2 fragments were produced by recombinant technology or by cleavage with Ficin. The scFv was tailored by use of the variable domain genes of the light and heavy chain from the hybridoma clone in combination with a (Gly4Ser)3-linker, followed by expression in E. coli. When tested for clearing capacity, the intact divalent antiidiotypic IgG was found to be the most efficient. The divalent Fab'2 and the monovalent Fab fragment also demonstrated significant clearing, but lower than the intact antiidiotypic IgG. The anti-TS1 scFv antibody when injected separately was not found to clear the idiotype, but could do so when preincubated with the idiotype. Rapid excretion and in vivo instability of this lowmolecular weight antibody fragment may be the major reasons. Similar results were obtained when the system was reversed and the 131I-labeled antiidiotype IgG was cleared with the idiotype Fab'2 fragment. It is concluded that both intact antiidiotypic IgG, Fab'2 and Fab fragments are able to clear the idiotypic antibodies. The experimental data support the conclusion that the Fc parts from both the idiotype and the antiidiotype may contribute to this elimination