Gene therapy using HVJ-liposomes: the best of both worlds?

A new concept for the development of novel vectors is to overcome the limitations of individual vectors by combining them. The HVJ-liposome was developed by combining liposomes with fusion proteins derived from the hemagglutinating virus of Japan (HVJ), also known as Sendai virus. Gene transfer in vivo using this delivery system can be repeated because it is much less immunogenic and cytotoxic than other viral-vector systems. By coupling the Epstein-Barr virus (EBV) replicon apparatus with HVJ-liposomes, transgene expression can be sustained in vitro and in vivo. In animal models, this system has shown promise for several diseases, including cancer and cardiovascular disease.     

Rikkunshi-to attenuates adverse gastrointestinal symptoms induced by fluvoxamine

Background  

The α-EEG anomaly during sleep, originally associated with chronic pain, is noted in several psychiatric and medical conditions and is also present in some normal subjects. The exact significance of the α-EEG anomaly is uncertain, but it has been suggested to be a nonspecific response to a variety of noxious stimuli. We propose that attachment insecurity, which is often associated with a state of hypervigilance during wakefulness, may be associated with the α-EEG anomaly during sleep.     

Alteration of the cell surface acid phosphatase concomitant with the morphological transformation in Trypanosoma cruzi

Acid phosphatase activity in Trypanosoma cruzi was found to be located on the external surface of the plasma membranes. Both specific activity and activity per cell of this bound enzyme were significantly higher in the cells of amastigote (an intracellular form) than that of trypomastigote (a bloodstream form) and epimastigote (culture form). During the transformation of epimastigotes to amastigotes in vitro the activity of surface acid phosphatase was elevated concomitant with the increase in population of amastigotes. These results were interpreted as that the elevated enzyme activity is required for the intracellular parasitization of this organism or is a consequence of the morphological transformation.     

Cloning and sequence of a functionally active cDNA encoding the mouse ubiquitin-activating enzyme E1.

A cDNA encoding the ubiquitin-activating enzyme, E1, was isolated from the mouse mammary carcinoma cell line, FM3A, and shown to complement mutant mouse cells deficient in the enzyme. The 3495-bp cDNA encodes 1058 amino acids (aa), and shares extensive homology with the human E1 enzyme at both the nucleotide and aa sequence levels.     

Detachment Activity but Not Cytotoxicity Induced in a T-Cell Clone Following Antigen Presentation in the Presence of Thymic Epithelial Cells

The selective induction of effector functions of a T-cell clone (DB14), specific to pigeon cytochrome c 43–58 (p 43–58) and restricted to I-A^b, was analyzed using a professional antigen-presenting cell, B hybridoma (Th 2.58), and various non-professional antigen-presenting cells (APC), L cells transfected with I-A^b (I-A^b L cells), a medullary thymic epithelial cell line (m-TEC) and a cortical thymic epithelial cell line (c-TEC). The m-TEC and c-TEC expressed I-A^b upon induction with interferon γ (IFN-γ). When stimulated with p 43–58 in the presence of I-A^b L cells as well as Th 2.58 cells, the DB14 cells showed marked proliferation and, after 18 hr of culturing, exhibited significant cytotoxicity against the APC. By contrast, in the presence of m, c-TEC, the DB14 cells showed neither proliferation nor cytotoxicity against these TEC but exhibited considerable detachment activity towards them. Furthermore, DB14 cells became expressed activation markers (CD69 or CD44) following stimulation with p 43–58 plus m-TEC or c-TEC. The addition of rIL-2 to the culture of DB14 cells, p 43–58 and m-TEC or c-TEC, restored the proliferative responses. However, it was shown that anergy was not involved in the negligible proliferative responses of DB14 cells after stimulation with p 43–58 plus m, c-TEC. The present findings indicate that differences in APC functions are present among non-professional APC and suggest that the selective induction of T-cell functions can be achieved using the appropriate non-professional APC. The characteristic activation of T cells by TEC may be related to their functional roles in situ.     

Secretion of Mr 46,000 protein from human hepatoma cells treated with tumor promotors is regulated by c-myc gene expression

Previously an Mr 46,000 protein (named p46) was shown to be induced in the culture medium of human hepatoblastoma cells, Huh-6 Cl-5, treated with 12-O-tetradecanoyl phorbol-13-acetate (TPA). For further characterization of p46, Huh-7 Cl-4, another line of well differentiated liver cells, was incubated in the presence and absence of TPA, and proteins were labeled with [35S]methionine, and the proteins secreted into the medium were analyzed by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. A protein of Mr 46,000 was observed in the culture medium of Huh-7 Cl-4 cells treated by TPA or plated at low density, but only slightly in the culture medium of Huh-7 Cl-4 cells plated at high density. As these results suggested that the expression of p46 was regulated by a factor that was activated in the growth state and by TPA treatment, the effect of introduction of the competence gene c-myc into Huh-7 Cl-4 cells was examined. All 37 independent transfected colonies obtained showed enhanced expression of p46. As a control, the c-Ha-ras gene was introduced into Huh-7 Cl-4 cells and shown not to enhance expression of p46. These results strongly suggested that the expression of p46 was regulated by the competence gene c-myc.