Erucylphosphocholine-induced apoptosis in glioma cells: involvement of death receptor signalling and caspase activation

Erucylphosphocholine (ErPC) is a promising anti-neoplastic drug for the treatment of malignant brain tumours. It exerts strong anti-cancer activity in vivo and in vitro and induces apoptosis even in chemoresistant glioma cell lines. The purpose of this study was to expand on our previous observations on the potential mechanisms of ErPC-mediated apoptosis with a focus on death receptor activation and the caspase network. A172 and T98G glioma cells were treated with ErPC for up to 48 h. ErPC effects on the expression of the tumour necrosis factor (TNF) and TNF-related apoptosis-inducing ligand (TRAIL) receptor system, and on caspase activation were determined. ErPC had no effect on the expression of TNFalpha or TRAIL. Inhibition of the TNF or TRAIL signalling pathway with antagonistic antibodies or fusion proteins did not affect apoptosis induced by ErPC, and a dominant-negative FADD construct did not abolish ErPC-induced effects. Western blot analysis indicated that ErPC-triggered apoptosis resulted in a time-dependent processing of caspases-3, -7, -8 and -9 into their respective active subunits. Co-treatment of A172 cells with different caspase inhibitors prevented apoptosis but did not abrogate cell death. These data suggest that A172 cells might have an additional caspase-independent pathway that insures cell death and guarantees killing of those tumour cells whose caspase pathway is incomplete.     

Contrasting IgG structures reveal extreme asymmetry and flexibility

The crystal structure of IgG1 b12 represents the first visualization of an intact human IgG with a full-length hinge that has all domains ordered and visible. In comparison to intact murine antibodies and hinge-deletant human antibodies, b12 reveals extreme asymmetry, indicative of the extraordinary interdomain flexibility within an antibody. In addition, the structure provides an illustration of the human IgG1 hinge in its entirety and of asymmetry in the composition of the carbohydrate attached to each C(H)2 domain of the Fc. The two separate hinges assume different conformations in order to accommodate the vastly different placements of the two Fab domains relative to the Fc domain. Interestingly, only one of two possible intra-hinge disulfides is formed.     

Molecular biology of nickel carcinogenesis

A review of the molecular mechanisms of nickel carcinogenesis has been compiled. This work is based upon approximately 20 years of research conducted in my laboratory. Molecular mechanisms of nickel carcinogenesis are considered from the pointofview of the uptake of nickel, both soluble and insoluble particles in cells, its dissolution and its effects on heterochromatin. Molecular mechanisms by which nickel induces gene silencing in cells by DNA hypermethylation in mammalian cells and by inhibiting histone acetylation in yeast cells are also discussed.     

Involvement of Erks activation in cadmium‐induced AP‐1 transactivation in vitro and in vivo

Cadmium is a potent and effective carcinogen in rodents and has recently been accepted by IARC (International Agency for Research on Cancer) as a category 1 carcinogen. Cadmium-induced upregulation of intracellular signaling pathways leading to increased mitogenesis is thought to be a major mechanism for the carcinogenic activity following chronic cadmium exposure. In the present study, we found that exposure of cells to cadmium induced significant activation of AP1 and all three members of the MAP kinase family in mouse epidermal JB6 cells. The induction of AP1 activity by cadmium appears to involve activation of Erks, since the induction of AP1 activity by cadmium was blocked by pretreatment of cells with PD98058. Interestingly, the induction of AP1 by cadmium was greatly enhanced by the chemical tumor promoter, TPA and the growth factor EGF, but not by ultraviolet C radiation. In vivo studies demonstrated that cadmium could also induce transactivation of AP1 in AP1luciferase report transgenic mice. Considering the role of AP1 activation in tumor promotion, the results presented in this study provide a possible molecular mechanism for cadmiuminduced carcinogenesis.     

Growth rates of Salvinia molesta in Lake Moondarra,Mount Isa,Australia

The seasonal growth of Salvinia molesta Mitchell was recorded during the period March 1978 to April 1979 in floating quadrats established in Lake Moondarra and a nearby sewage lagoon. Primary-form plants were used as inoculum in the quadrats; leaf numbers, fresh weight and percentage cover changes were recorded. These three parameters were significantly correlated. The highest individual rates of increase recorded in the lagoon were 51.4% per day for percentage cover, 50.1% per day for leaf numbers and 38.3% per day for fresh weight. In the lake, the highest values were 19.0% per day, 25.7% per day and 16.6% per day, respectively. A simple growth model for this species, relating growth rate to water temperature, accounted for 76% of the observed variance in leaf number growth rates in the lagoon. The same model, however, only explained 5% of the variance in the lake, suggesting growth limitations by other factors.     

Instructed heart rate control in the presence and absence of a distracting task: The effects of biofeedback training

Twenty volunteers participated in a single-session experiment in which bidirectional heart rate (HR) control was assessed before and after brief unidirectional HR biofeedback. Subjects attempted to raise (INC) and lower (DEC) HR while performing mental arithmetic, as well as in no-task conditions. Biofeedback training was also carried out in the presence and absence of mental arithmetic. Subjects were divided into two groups on the basis of initial HR reactivity to mental arithmetic. Group U received feedback and instructions to raise HR during the training period, while group D attempted to lower HR. Significant differences in HR modifications during INC and DEC trials were observed prior to any biofeedback training in no-task conditions. Following training, however, ability to raise HR deteriorated in group D, while HR decelerations were impaired in group U. Unidirectional training in HR control thus handicapped subsequent attempts to modify HR in the reverse direction. The pattern of HR change was generally paralleled by respiration rate. Subjects were also able to influence the cardiac reactions to mental arithmetic even before the administration of biofeedback. The data nevertheless suggest that training affects the magnitude of HR reactions after the biofeedback is withdrawn. In the biofeedback phase itself, the HR increases and decreases produced by groups U and D, respectively, were diminished on simultaneous mental arithmetic performance.The authors are grateful to Drs. Beryl Starr and Alvin Ross for their advice at various stages of this project.     

Identification of Group A Streptococci by Direct Fluorometry

A simple direct fluorometric method for rapid identification of group A streptococci is described. The method permits the detection of the organism in mixed cultures without the aid of a microscope and is amenable to automated processing of specimens. Experience with the indirect fluorometric method revealed that nontrypsinized cells from a 10-fold dilution of overnight broth cultures could be stained with uniform brilliance with fluorescent antibody (1:15 dilution) and that fluorescent antibody dissociated from such cells at 55 C for 20 min gave serologically specific fluorometric values. With this information, it was possible to develop a simpler fluorometric test which gave results comparable to those obtained by conventional cultural-precipitin grouping techniques. In the direct test described, cultures from throat swabs were incubated overnight, and cells from a 10-fold dilution were stained with specific fluorescent antibody (1:50 dilution) and then rinsed. The stained specimens were transferred to a continuous-filter paper strip (Whatman 3 MM) and read serially in a Turner 110 fluorometer with Corning 5840 and Wratten 2A filters in place. The reagents used required careful standardization and testing to assure that fluorometric readings above a specified value would be indicative of the presence of group A streptococci.