Mitochondriome and Cholangiocellular Carcinoma

Cholangiocellular carcinoma (CCA) of the liver was the target of more interest, recently, due mainly to its increased incidence and possible association to new environmental factors. Somatic mitochondrial DNA (mtDNA) mutations have been found in several cancers. Some of these malignancies contain changes of mtDNA, which are not or, very rarely, found in the mtDNA databases. In terms of evolutionary genetics and oncology, these data are extremely interesting and may be considered a sign of poor fitness, which may conduct in some way to different cellular processes, including carcinogenesis. MitoChip analysis is a strong tool for investigations in experimental oncology and was carried out on three CCA cell lines (HuCCT1, Huh-28 and OZ) with different outcome in human and a Papova-immortalized normal hepatocyte cell line (THLE-3). Real time quantitative PCR, western blot analysis, transmission electron microscopy, confocal laser microscopy, and metabolic assays including L-Lactate and NAD⁺/NADH assays were meticulously used to identify mtDNA copy number, oxidative phosphorylation (OXPHOS) content, ultrastructural morphology, mitochondrial membrane potential (ΔΨ_m), and differential composition of metabolites, respectively. Among 102 mtDNA changes observed in the CCA cell lines, 28 were non-synonymous coding region alterations resulting in an amino acid change. Thirty-eight were synonymous and 30 involved ribosomal RNA (rRNA) and transfer RNA (tRNA) regions. We found three new heteroplasmic mutations in two CCA cell lines (HuCCT1 and Huh-28). Interestingly, mtDNA copy number was decreased in all three CCA cell lines, while complexes I and III were decreased with depolarization of mitochondria. L-Lactate and NAD+/NADH assays were increased in all three CCA cell lines. MtDNA alterations seem to be a common event in CCA. This is the first study using MitoChip analysis with comprehensive metabolic studies in CCA cell lines potentially creating a platform for future studies on the interactions between normal and neoplastic cells.     

An ethogram for rhesus monkeys I. Antithetical contrasts in posture and movement

Antithetical postures and movements in social displays of rhesus monkeys distinguish five moods. Movement from a high to a low position in the midsagittal plane indicates threat. Emphasis on movement in the frontal plane indicates subordinance. Movement from a low to a high position in the midsagittal plane indicates non-hostile, non-fearful approach. Oblique movements indicate a querying mood. Playfulness is indicated by emphasis on rotatory movements in the transverse plane.     

Effects of oestradiol, the oestrous cycle and pregnancy on weight, metabolism and cytosol receptors in the oviduct and vaginal complex of the brushtail possum (Trichosurus vulpecula)

Weight, RNA, DNA and protein content of the oviduct, vaginal cul-de-sac, lateral vagina and urogenital sinus and oestradiol and progesterone cytosol receptor concentrations in vaginal cul-de-sac, lateral vagina and urogenital sinus were examined after administration of oestradiol to ovariectomized animals and on days 0, 5, 9 and 13 of the non-pregnant cycle and on day 13 of the pregnant cycle. In ovariectomized animals, oestradiol induced an increase in weight, RNA:DNA and protein:DNA ratios and a decrease in DNA:tissue weight ratio for each organ and in addition an increase in total DNA in vaginal cul-de-sac and urogenital sinus. There was no effect of oestradiol on oestradiol cytosol receptor concentration but there was a significant increase in progesterone cytosol receptor concentration in all organs that were examined. During the oestrous cycle, changes in the wet weight of each organ showed a common pattern with maximum weight at day 0 followed by variable rates of decline until day 13. In oviduct and vaginal cul-de-sac, the decrease in weight was paralleled by a decrease in RNA:DNA and protein:DNA ratios whereas the DNA: tissue weight ratio showed the opposite pattern and total DNA remained unchanged. The changes in the lateral vagina and urogenital sinus were similar except that a significant decline in total DNA was also seen after day 0 and the DNA:tissue weight and protein:DNA ratios in the urogenital sinus and the lateral vagina respectively showed no significant changes. Progesterone cytosol receptor concentration in the lateral vagina and urogenital sinus were high on day 0 and then declined until day 13. In contrast, there were no consistent effects on oestradiol receptor concentration.     

Effector domain mutations dissociate p21ras effector function and GTPase-activating protein interaction.

The GTPase activity of p21ras is stimulated by GTPase-activating proteins (GAPs) such as p120GAP and the product of the neurofibromatosis 1 gene, which may negatively regulate p21 function. GAPs are also proposed effectors of ras. We have sought activating substitutions in c-H-ras in the region encoding the effector domain, on the rationale that such mutations would dissociate effector function from negative regulation by GAP. One such activating mutation, Pro-34-->Arg, encodes protein that is substantially bound to GTP in vivo. In vitro, this protein is not stimulated by GAPs, and its binding to p120GAP is grossly impaired. The results support the idea that the p21 structural requirements for effector function and GAP interaction are quite different and suggest that some molecule(s) other than p120GAP serves as the ras effector. In contrast to the results obtained with p120GAP, the Pro-34-->Arg p21 species is effectively coupled to the raf-1 product, as judged from electrophoretic mobility shifts of the Raf-1 phosphoprotein.     

Geochemical composition,phosphorus speciation and mass transport of fine-grained sediment in two Lake Erie tributaries

The concentration of major elements (Si, Al, Ca, Mg, Na, K, Fe, Ti, Mn and P), particulate phosphorus forms (NH₄Cl-RP, BD-RP, NaOH-RP, HCl-RP and NaOH₍₈₅₎-RP) and carbon content were determined in six size fractions (<8, 8–12, 12–19, 19–31, 31–42 and 42–<60 µm) of sediment collected at gauging stations located in two Lake Erie tributaries (Big Creek and Big Otter Creek). Concentrations of major elements and phosphorus forms were remarkably similar in sediment size fractions from both rivers. Nonapatite inorganic P (NAIP) and organic P (OP) concentrations increased with decreasing grain size while apatite inorganic P (AIP) content decreased with decreasing grain size. Results of phosphorus fractionation studies were combined with historical (particle size) and hydrometric data to simulate the export of particle P on tributary sediment < 63 µm. AIP represents 67 and 70% of the calculated particulate P mass while NAIP accounts for 26 and 23% of sediment-bound P transported in Big Otter Creek and Big Creek, respectively. The < 8 µm size fraction of tributary sediment is the most significant for the potential release of bioavailable P into the water column.