Morphologic and functional peculiarities of the visceral protoneuron reexamined in light of recent data

The recent works showing the richness and complexity of the sensory innervation of viscera raise various questions which are of general interest for modern physiology. 1 - The primary afferent neurons are much more numerous and heterogeneous than it was usually expected. Moreover, another category of neurones, namely the sensory intrinsic neurons, must be taken in consideration. These neurones end either in the intrinsic plexuses of viscera or in the prevertebral ganglia. 2 - Another major question concerns the mechanisms responsible for the functional diversity of interoceptors including the chemoreceptors. Several hypothesis have been proposed involving the presence of membrane receptors, the release of neurotransmitters by chromaffin cells and the deformation of tissue. In any way, the functional characteristics of interoceptors do not result in morphological differentiation since free endings are mainly found in the visceral area. 3 - Does the functional properties of primary afferent neurons are related to their morphological (diameter of fibres, size of cell bodies) or their biochemical (type of neurotransmitters) characteristics? So far, it is not possible to answer this question. Nevertheless the available data suggest that a such relationship partly exists. 4 - The visceral afferents are largely implicated in various physiological mechanisms involving visceral motility, homeostasis and behaviour. Therefore, their role in pain appear relatively less important than previously expected. Conversely, the nervous mechanisms induced by the interoceptor activation play a major part in that they prepare and potentialize the effects produced by the other mechanisms.     

Multiple effects of differentiation-inducing factor on prespore differentiation and cyclic-AMP signal transduction in Dictyostelium

Abstract. The effects of the differentiation inducing factor (DIF) on several CAMP-induced responses in Dictyostelium were investigated. It was found that DIF reduces the apparent affinity of cell-surface cAMP receptors. DIF does not affect the CAMP-induced cGMP response, but it is a potent inhibitor of the CAMP-relay response. DIF also inhibits the induction of prespore differentiation by cAMP in aggregation-competent cells. We also compared the effects of DIF on CAMP-induced responses with those of the relay inhibitor, caffeine, and the morphogen, adenosine.     

Autophagy induction by xanthoangelol exhibits anti‐metastatic activities in hepatocellular carcinoma

Xanthoangelol (XAG), a prenylated chalcone isolated from the Japanese herb Angelica keiskei Koidzumi, has been reported to exhibit antineoplastic properties. However, the specific anti‐tumor activity of XAG in human hepatocellular carcinoma (HCC), and the relevant mechanisms are not known. Herein, we evaluated the effect of XAG against HCC in vitro and in vivo. Although XAG treatment did not significantly reduce the viability of the Hep3B and Huh7 cell lines, it suppressed cell migration, invasion, and EMT. This anti‐metastatic effect of XAG was due to induction of autophagy, because treatment with the autophagy inhibitor 3‐methyadenine (3‐MA) or knockdown of the pro‐autophagy Beclin‐1 effectively abrogated the XAG‐induced suppression of metastasis. Mechanistically, XAG induced autophagy via activation of the AMPK/mTOR signaling pathway, and XAG treatment dramatically increased the expression of p‐AMPK while decreasing p‐mTOR expression. In addition, blocking AMPK/mTOR axis with compound C abrogated the autophagy‐mediated inhibition of metastasis. The murine model of HCC metastasis also showed that XAG effectively reduced the number of metastatic pulmonary nodules. Taken together, our results revealed that autophagy via the activation of AMPK/mTOR pathway is essential for the anti‐metastatic effect of XAG against HCC. These findings not only contribute to our understanding of the anti‐tumor activity of XAG but also provide a basis for its clinical application in HCC. Before this study, evidence of XAG on HCC was purely anecdotal; present study provides the first comprehensive assessments of XAG on HCC metastasis and investigates its underlying mechanism. Results suggest that XAG exerts anti‐metastatic properties against HCC through inducing autophagy which is mediated by the activation of AMPK/mTOR signaling pathway. This research extends our knowledge about the antineoplastic properties of XAG and suggests that induction autophagy may represent future treatment strategies for metastatic HCC.     

Lectin Binding Assays for In-Process Monitoring of Sialylation in Protein Production

Many therapeutic proteins require appropriate glycosylation for their biological activities and plasma half life. Coagulation factor VIII (FVIII) is a glycoprotein which has extensive post-translational modification by N-linked glycosylation. The terminal sialic acid in the N-linked glycans of FVIII is required for maximal circulatory half life. The extent of FVIII sialylation can be determined by high pH anion-exchange chromatography coupled with a pulse electrochemical detector (HPAEC-PED), but this requires a large amount of purified protein. Using FVIII as a model, the objective of the present study was to develop assays that enable detection and prediction of sialylation deficiency at an early stage in the process and thus prevent downstream product quality excursions. Lectin ECA (Erythrina Cristagalli) binds to unsialylated Galβ1-4 GlcNAc and the ECA-binding level (i.e., terminal Gal(β1-4) exposure) is inversely proportional to the level of sialylation. By using ECA, a cell-based assay was developed to measure the global sialylation profile in FVIII producing cells. To examine the Galβ1-4 exposure on the FVIII molecule in bioreactor tissue culture fluid (TCF), an ELISA-based ECA-FVIII binding assay was developed. The ECA-binding specificity in both assays was assessed by ECA-specific sugar inhibitors and neuraminidase digestion. The ECA-binding specificity was also independently confirmed by a ST3GAL4 siRNA knockdown experiment. To establish the correlation between Galβ1-4 exposure and the HPAEC-PED determined FVIII sialylation value, the FVIII containing bioreactor TCF and the purified FVIII samples were tested with ECA ELISA binding assay. The results indicated an inverse correlation between ECA binding and the corresponding HPAEC-PED sialylation value. The ECA-binding assays are cost effective and can be rapidly performed, thereby making them effective for in-process monitoring of protein sialylation.     

Electrophysiology of the peripheral effect of two analgesics: aspirin and dibencozide

The peripheral effect of two analgesics (aspirin and dibencozide) was studied on anaesthetized cats. Several types of neurons and stimulations were performed in this work: traction for periodontal mechanoreceptors connected to small-sized trigeminal fibres, distension for the muscular intestinal mechanoreceptors connected to non-myelinated vagal fibres, chemical stimulation by means of phenyldiguanide for the non-myelinated vagal fibres, electrical stimulation of the myelinated and non-myelinated vagal fibres. In all cases, unitary activities were recorded into corresponding ganglia (nodose or gasserian) with extracellular glass microelectrodes. After injection of analgesics, a decrease of control responses were observed till 30 minutes but the maximum occurred between 1 and 5 minutes. This effect concerned the non-myelinated neurones as well as the myelinated ones. It can be explained by a direct action of analgesics on the ending excitability.