Determinants of interactions of a novel next-generation gabapentinoid NVA1309 and mirogabalin with the Cavα2δ-1 subunit.

NVA1309 is a non-brain penetrant next-generation gabapentinoid shown to bind Cavα2δ at R243 within a triple Arginine motif forming the binding site for gabapentin and pregabalin. In this study we have compared the effects of NVA1309 with Mirogabalin, a gabapentinoid drug with higher affinity for the voltage-gated calcium channel subunit Cavα2δ-1 than pregabalin which is approved for post-herpetic neuralgia in Japan, Korea and Taiwan. Both NVA1309 and mirogabalin inhibit Cav2.

A next generation peripherally restricted Cavα2δ-1 ligand with inhibitory action on Cav2.2 channels and utility in neuropathic pain.

The Voltage-Gated Calcium Channel (VGCC) auxiliary subunit Cavα2δ-1 (CACNA2D1) is the target/receptor of gabapentinoids which are known therapeutics in epilepsy and neuropathic pain. Following damage to the peripheral sensory nervous system, Cavα2δ-1 is upregulated in dorsal root ganglion (DRG) neurons in several animal models of chronic neuropathic pain. Gabapentinoids, such as gabapentin and pregabalin, engage with Cavα2δ-1 via binding an arginine residue (R241) within an RRR motif located at the N-terminus of human Cavα2δ-1.

The effect of activated carbon addition on membrane bioreactor processes for wastewater treatment and reclamation - A critical review.

This review concentrates on the effect of activated carbon (AC) addition to membrane bioreactors (MBRs) treating wastewaters. Use of AC-assisted MBRs combines adsorption, biodegradation and membrane filtration. This can lead to advanced removal of recalcitrant pollutants and mitigation of membrane fouling.

Intramedullary nailing for the treatment of aseptic femoral shaft non-unions after plating failure: effectiveness and timing.

This retrospective, multicentre study aimed to evaluate reamed intramedullary nailing (IMN) for the treatment of 30 cases of aseptic femoral shaft non-union after plating failure. Following nailing, 29 non-unions had healed by a mean 7.93 months.

Hepatocyte Growth Factor/scatter factor-induces phosphorylation of cortactin in A431 cells in a Src kinase-independent manner.

The Hepatocyte Growth Factor receptor transduces proliferating and scattering signals in epithelial and endothelial cells. We have explored potential interactions of the HGF/SF receptor beta-subunit (p145(beta MET)) with F-actin binding partners aiming to identify novel downstream effectors implicated in HGF/SF pluripotent signalling. Cortactin, a p80/85 F-actin binding protein, was found phosphorylated on tyrosine in response to HGF-SF in A431 human epidermoid carcinoma cells, expressing the HGF/SF receptor (c-MET).

The beta-subunit of the hepatocyte growth factor/scatter factor (HGF/SF) receptor phosphorylates and associates with CrkII: expression of CrkII enhances HGF/SF-induced mitogenesis.

CrkII, a 40 kDa adaptor possessing a Src homology (SH)2 domain followed by two SH3 domains, although not endowed with catalytic activity, participates in intracellular signalling, presumably by activating the Ras pathway. CrkII was found to be phosphorylated in response to hepatocyte growth factor/scatter factor (HGF/SF) and to associate with the beta-subunit of the HGF receptor (MET). CrkII associated with p(145betaMET) via its SH2 domain.

Hepatocyte growth factor/scatter factor-induced intracellular signalling.

Hepatocyte growth factor (HGF) identical to scatter factor (SF) is a glycoprotein involved in the development of a number of cellular phenotypes, including proliferation, mitogenesis, formation of branching tubules and, in the case of tumour cells, invasion and metastasis. This fascinating cytokine transduces its activities via its receptor encoded by the c-met oncogene, coupled to a number of transducers integrating the HGF/SF signal to the cytosol and the nucleus. The downstream transducers coupled to HGF/MET, most of which participate in overlapping pathways, determine the development of the cell's phenotype, which in most cell types is dual.

Cytosolic phospholipase A2 is activated by the hepatocyte growth factor receptor-kinase in Madin Darby canine kidney cells.

The hepatocyte growth factor/scatter factor (HGF/SF) receptor which is a transmembrane protein encoded by the Met oncogene, possesses intrinsic tyrosine kinase activity which transduces the mitogenic, morphogenic and the scattering effect of HGF/SF. The pluripotent signal of HGF/SF is transduced through association of the Met receptor with various intracellular adaptors. Phosphorylation of cytosolic phospholipase A2 (cPLA2) is associated with activation of this molecule which in turn leads to arachidonic acid production followed by release of prostaglandins and related compounds exerting their roles onto cell proliferation, chemotaxis and vascular motility.

Inhibition of bacterial growth by synthetic SP-B1-78 peptides.

Residues 12-34 of mature human pulmonary surfactant protein B (SP-B1-78) are 68% homologous to residues 48-72 of the frog peptide antibiotic dermaseptin b I. We examined the effects of SP-B1-78 on the growth of Escherichia coli in order to find whether full length SP-B1-78 might act as a peptide antibiotic. We found that SP-B1-78 peptide inhibited growth of E.

The hepatocyte growth factor receptor kinase-mediated phosphorylation of lipocortin-1 transduces the proliferating signal of the hepatocyte growth factor.

Hepatocyte growth factor (HGF), which is identical to scatter factor (SF) through coupling to its receptor the product of c-met oncogene, was found to induce proliferation of A549 lung carcinoma cell line, accompanied by release of prostaglandin E2 (PGE2). This activity was sensitive to 0.1-100 microM indomethacin and to 5-50 nM of verapamil.

Parathyroid hormone co-stimulation of hepatocyte proliferation is sensitive to protein kinase A and calcium channel inhibitors.

Parathyroid hormone (PTH) mobilises calcium in the hepatocyte, an effect which is abolished by verapamil and staurosporine. In our study parathyroid hormone was shown to act additively to dHGF in inducing hepatocyte DNA synthesis. It is also shown that PTH induced the production of inositol 1,4,5 trisphosphate (IP3) and c-fos expression at early times in culture.

C-myc is required for the G0/G1-S transition of primary hepatocytes stimulated with a deleted form of hepatocyte growth factor.

Primary rat hepatocytes stimulated in vitro with the addition of a deleted form of hepatocyte growth factor (dHGF) enter the S-phase 48 h after addition of the growth factor. The c-myc gene is believed to play a role in a variety of cellular stages, such as proliferation, differentiation and cell death. In primary hepatocytes c-myc was expressed constitutively at both mRNA and protein levels, independently of the growth conditions.

c-Myc and Max interactions in quiescent and mitogen-stimulated primary hepatocytes.

The c-myc oncogene has been linked with cell proliferation, apoptosis, and differentiation, and when its expression is deregulated also with malignant transformation. In primary hepatocytes c-myc expression is constitutive and in part regulated by hepatocyte-specific growth factors (HGF, TGFalpha, and EGF) in a delayed early response manner. Max expression in these cells was found to be constitutive throughout the in vitro lifetime and mRNA transcript levels were increased at 12 h after induction with growth factors.

Hepatocyte growth factor-induced proliferation of primary hepatocytes is mediated by activation of phosphatidylinositol 3-kinase.

Primary hepatocytes respond to the proliferating signals of Hepatocyte Growth Factor (HGF) through activation of the tyrosine kinase activity of the met (p145) receptor. Addition of dHGF in hepatocyte cultures resulted in receptor phosphorylation which co-precipitated with a phosphorylated protein of 85 kDa. This protein was identified as the regulatory subunit of phosphatidylinositol 3-kinase (PI 3-kinase).

Phospholipase A2 is activated by tumor necrosis factor-alpha in primary hepatocytes stimulated by a deleted form of hepatocyte growth factor.

Several factors are released in the liver microenvironment immediately after injury. Among these factors TNF alpha is implicated as a regulator of hepatocyte proliferation. Hepatocytes in the intact liver are mostly in the G0 phase of the cell cycle and after injury (including collagenase perfusion) display a constitutive expression of the growth-regulated c-myc oncogene.

Expression of exogenous c-myc oncogene does not initiate DNA synthesis in primary rat hepatocyte cultures.

Cultured hepatocytes from adult rats stimulated with combinations of growth factors enter into S phase but do not undergo multiple rounds of DNA synthesis nor mitosis. We have examined the potential of an introduced oncogene to induce alterations in the DNA synthetic activity of the cultured hepatocytes in response to epidermal growth factor (EGF). Overexpression of c-myc did not initiate significant DNA synthesis in rat hepatocyte cultures alone, although it cooperated with added EGF to super-induce thymidine incorporation into DNA.

Transforming growth factor-alpha-induced DNA synthesis and c-myc expression in primary rat hepatocyte cultures is modulated by indomethacin.

Primary hepatocytes stimulated with epidermal growth factor (EGF) secrete prostaglandins into the culture medium as soon as 1 h after the addition of the EGF. Transforming growth factor-alpha (TGF alpha), a potent hepatocyte mitogen, shares the same receptor with EGF, and its expression is increased after partial hepatectomy. TGF alpha is also secreted in culture.

Synthesis and phosphorylation of an extracellular polypeptide reacting with anti-MYC antisera in primary rat hepatocyte cultures.

Primary hepatocytes stimulated with appropriate growth factors enter into S phase and it is believed that c-myc and other cell cycle-related genes play an important role in the G0-G1/S phase transition. Four polypeptides reacting with anti-MYC antisera were detected in normal primary rat hepatocyte lysates, showing a pattern of 55-67 KDa on SDS-PAGE. A 67 KDa polypeptide was detected in the extracellular medium of the hepatocyte culture capable of undergoing phosphorylation.

Hepatocyte conditioned medium modulates the response of primary rat hepatocyte cultures to epidermal growth factor.

Primary hepatocytes may produce autocrine growth trigger(s) with or without a mitogenic stimulus. We explored the potential of hepatocyte conditioned medium--from untreated quiescent cultures--to modulate the DNA synthetic responses induced by EGF. The EGF-induced responses were similar when EGF was continuously or transiently (3 h) present.

Prostaglandins E2 and F2a mediate the increase in c-myc expression induced by EGF in primary rat hepatocyte cultures.

Epidermal Growth Factor (EGF) and prostaglandins (PGs) E2 and F2a, have been shown to stimulate primary hepatocyte proliferation. Verapamil (5-20 microM), a calcium channel inhibitor, inhibited hepatocyte DNA synthesis and c-myc expression, induced by EGF (50 ng/dish) and prostaglandins (1-12 micrograms/dish). Indomethacin (20-100 microM) decreased significantly the EGF-induced hepatocyte DNA synthesis and c-myc expression.

A growth factor for hepatocytes is inactivated by sub-lethal gamma-irradiation in vivo or in vitro.

Serum from partially hepatectomized rats promoted DNA synthesis in primary adult rat hepatocyte cultures. If the rats had been exposed to sub-lethal gamma-irradiation immediately following operation or if their serum, collected at 3 h, was exposed to irradiation in vitro, the growth-promoting activity was destroyed. Prostaglandin E2 also stimulated DNA synthesis in the cultures; if PGE2 was irradiated in serum from intact or partially hepatectomized rats its growth-promoting activity was markedly diminished.

Serum from partially hepatectomized rats induces primary hepatocytes to enter S phase: a role for prostaglandins?

Serum obtained from partially hepatectomized rats 1, 3 or 24 h after operation was more effective in stimulating DNA synthesis in primary adult rat hepatocytes than serum from sham-operated rats; exposure to the serum for 2 h was sufficient to promote growth. Serum from the partially hepatectomized rats contained elevated levels of PGE2 and PGF2 alpha; it promoted hepatocytes to release prostaglandins into their culture medium. Growth-promoting effects of the serum and its capacity to elicit prostaglandin release into the culture medium were inhibited by 0.

Regulation of the proliferation of primary rat hepatocytes by eicosanoids.

DNA synthesis in primary adult rat hepatocyte cultures was promoted by epidermal growth factor (EGF), arachidonic acid, and prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha). Growth promotion by EGF was blocked by 0.1 mM indomethacin and 1 mM aspirin, without affecting cell viability.