Cells that overproduce protein kinase C are more susceptible to transformation by an activated H-ras oncogene.

We recently developed rat fibroblast cell lines that stably overproduce high levels of the beta 1 form of protein kinase C (PKC). These cells display several disorders in growth control and form small microscopic colonies in agar. In the present study we demonstrate that one of these cell lines, R6-PKC3, is extremely susceptible to transformation by an activated human bladder cancer c-H-ras oncogene (T24). Compared with control cell line R6-C1, T24-transfected R6-PKC3 cells yielded a 10-fold increase in the formation of large colonies in agar. Cell lines established from these colonies displayed a highly transformed morphology, expressed the T24-encoded p21 ras protein, continued to express high levels of PKC, and were highly tumorigenic in nude mice. These results provide genetic evidence that PKC mediates some of the effects of the c-H-ras oncogene on cell transformation. Data are also presented suggesting that optimum synergistic effects between c-H-ras and PKC require critical levels of their respective activities. These findings may be relevant to the process of multistage carcinogenesis in tissues containing cells with an activated c-H-ras oncogene.     

Evidence for altered expression of the GTP-dependent regulatory proteins, Gs and Gi, in adipocytes from aged rats.

The alpha-subunits of Gi and Gs were quantified in adipocyte membranes from young (2-month) and older (18-month) rats by pertussis-toxin and cholera-toxin labelling respectively. Aging was associated with a 3-fold increase in Gi alpha-subunit, but only a 2-fold increase in one of the two Gs alpha-subunit species labelled. The findings may explain the altered sensitivity of adipocytes from aged rats to lipolytic and anti-lipolytic stimuli.     

Characterization of a Lactobacillus strain producing white crystals on cheddar cheese

From an enrichment culture of white-crystal deposits from aged Cheddar cheese, an atypical Lactobacillus strain was characterized. The new isolate is facultatively heterofermentative, has a G + C content of 40 mol%, and produces D and L isomers of lactic acid. The strain had a limited ability to ferment carbohydrates. It utilized fructose, galactose, glucose, lactose, maltose, mannose, and ribose but was negative for esculin, gluconate, citrate, and several other carbon sources. The isolate also had low DNA-DNA homologies with strains of Lactobacillus casei and Lactobacillus plantarum. Cheese prepared with milk containing the isolate developed white crystals during curing. Formation of copious D-lactate from unknown substrates during curing probably caused the white-crystal deposits. The strain has been deposited in the American Type Culture Collection (ATCC 49178).     

Diffusion of the Interspecies Electron Carriers H(2) and Formate in Methanogenic Ecosystems and Its Implications in the Measurement of K(m) for H(2) or Formate Uptake

We calculated the potential H(2) and formate diffusion between microbes and found that at H(2) concentrations commonly found in nature, H(2) could not diffuse rapidly enough to dispersed methanogenic cells to account for the rate of methane synthesis but formate could. Our calculations were based on individual organisms dispersed in the medium, as supported by microscopic observations of butyrate-degrading cocultures. We isolated an axenic culture of Syntrophomonas wolfei and cultivated it on butyrate in syntrophic coculture with Methanobacterium formicicum; during growth the H(2) concentration was 63 nM (10.6 Pa). S. wolfei contained formate dehydrogenase activity (as does M. formicicum), which would allow interspecies formate transfer in that coculture. Thus, interspecies formate transfer may be the predominant mechanism of syntrophy. Our diffusion calculations also indicated that H(2) concentration at the cell surface of H(2)-consuming organisms was low but increased to approximately the bulk-fluid concentration at a distance of about 10 mum from the surface. Thus, routine estimation of kinetic parameters would greatly overestimate the K(m) for H(2) or formate.     

Exciton interactions in synthetic porphyrin dimers

The Soret absorption spectra of six synthetic rigid porphyrin dimers whose crystal structures have been determined are simulated using simple exciton theory. The objective is to test the validity of the point dipole and associated approximations; the electronic interaction parameters are thus calculated using data obtained from the monomer spectra, with no adjustable parameters. Satisfactory agreement between theory and experiment is obtained for one class of dimers but not for a second. This poses a challenge for semiempirical electronic structure methods as to whether improvements over the point dipole calculations can be obtained.     

Aliphatic hydrocarbons obtained from iron carbides: Possible stellar and meteoritic implications

Origins of Life and Evolution of Biospheres -     

Binding of a pancreatic nuclear protein is correlated with amylase enhancer activity.

The mouse amylase gene Amy-2.2 is expressed at high levels specifically in the acinar cells of the pancreas. The region between -172 and -110 of this gene includes sequence elements common to pancreas-specific genes. Nuclear proteins with specific affinity for this region were partially purified from rat pancreas. The consensus element of another pancreas-specific gene, elastase 1, competes for protein binding to the amylase sequences. Binding was localized by DNase I protection to the sequence -156 to -122. Site-directed mutagenesis of this sequence resulted in concomitant loss of protein binding and enhancer activity. Photo-affinity labelling of pancreatic nuclear extracts identified one predominant binding protein with a molecular weight of approximately 75 kDa. The data indicate that binding of this nuclear protein is essential for the enhancer activity of this pancreas-specific element.