Mitochondrial Inheritance Is Delayed in Saccharomyces cerevisiae Cells Lacking the Serine/Threonine Phosphatase PTC1

In wild-type yeast mitochondrial inheritance occurs early in the cell cycle concomitant with bud emergence. Cells lacking the PTC1 gene initially produce buds without a mitochondrial compartment; however, these buds later receive part of the mitochondrial network from the mother cell. Thus, the loss of PTC1 causes a delay, but not a complete block, in mitochondrial transport. PTC1 encodes a serine/threonine phosphatase in the high-osmolarity glycerol response (HOG) pathway. The mitochondrial inheritance delay in the ptc1 mutant is not attributable to changes in intracellular glycerol concentrations or defects in the organization of the actin cytoskeleton. Moreover, epistasis experiments with ptc1Δ and mutations in HOG pathway kinases reveal that PTC1 is not acting through the HOG pathway to control the timing of mitochondrial inheritance. Instead, PTC1 may be acting either directly or through a different signaling pathway to affect the mitochondrial transport machinery in the cell. These studies indicate that the timing of mitochondrial transport in wild-type cells is genetically controlled and provide new evidence that mitochondrial inheritance does not depend on a physical link between the mitochondrial network and the incipient bud site.     

Raloxifene and desmethylarzoxifene block estrogen-induced malignant transformation of human breast epithelial cells

There is association between exposure to estrogens and the development and progression of hormone-dependent gynecological cancers. Chemical carcinogenesis by catechol estrogens derived from oxidative metabolism is thought to contribute to breast cancer, yet exact mechanisms remain elusive. Malignant transformation was studied in MCF-10A human mammary epithelial cells, since estrogens are not proliferative in this cell line. The human and equine estrogen components of estrogen replacement therapy (ERT) and their catechol metabolites were studied, along with the influence of co-administration of selective estrogen receptor modulators (SERMs), raloxifene and desmethyl-arzoxifene (DMA), and histone deacetylase inhibitors. Transformation was induced by human estrogens, and selectively by the 4-OH catechol metabolite, and to a lesser extent by an equine estrogen metabolite. The observed estrogen-induced upregulation of CYP450 1B1 in estrogen receptor negative MCF-10A cells, was compatible with a causal role for 4-OH catechol estrogens, as was attenuated transformation by CYP450 inhibitors. Estrogen-induced malignant transformation was blocked by SERMs correlating with a reduction in formation of nucleobase catechol estrogen (NCE) adducts and formation of 8-oxo-dG. NCE adducts can be formed consequent to DNA abasic site formation, but NCE adducts were also observed on incubation of estrogen quinones with free nucleotides. These results suggest that NCE adducts may be a biomarker for cellular electrophilic stress, which together with 8-oxo-dG as a biomarker of oxidative stress correlate with malignant transformation induced by estrogen oxidative metabolites. The observed attenuation of transformation by SERMs correlated with these biomarkers and may also be of clinical significance in breast cancer chemoprevention.     

Raphidascaroides brasiliensis n. sp. (Nematoda: Anisakidae), an intestinal parasite of the thorny catfish Pterodoras granulosus from Amazonia,Brazil

A new anisakid nematode, Raphidascaroides brasiliensis n. sp., is described from the intestine of the freshwater thorny catfish Pterodoras granulosus (Valenciennes) (Doradidae, Siluriformes) from Amazonia (Manaus), Brazil. It is characterised mainly by the smooth, almost rounded tail tip in both sexes, the length of the spicules (0.952–1.183 mm) and by the number and arrangement of the caudal papillae (24–34 pre-anal, 1 adanal and 5 postanal pairs and 1 median pre-anal papilla) in the male. It is the first Raphidascaroides species described from South America and the second species of this genus reported from a freshwater fish.     

In vitro actions on hemopoietic cells of recombinant murine GM-CSF purified after production in Escherichia coli: comparison with purified native GM-CSF

Recombinant murine GM-CSF produced in Escherichia coli was purified to homogeneity and tested in parallel with purified native GM-CSF. Both recombinant and native GM-CSF stimulated granulocyte and/or macrophage colony formation by adult and fetal mouse progenitor cells, and with adult marrow cells the specific activity of the recombinant GM-CSF (25 X 10(8) U/mg) was similar to that of the native form (15 X 10(8) U/mg). At high concentrations (greater than 200 U/ml), both forms of GM-CSF also stimulated eosinophil colony formation by adult marrow cells and, at very high concentrations (greater than 800 U/ml), megakaryocyte and some erythroid and mixed-erythroid colony formation. Recombinant GM-CSF was as effective in stimulating the proliferation of the GM-CSF-dependent cell line FD as the native molecule. Both recombinant and native GM-CSF were able to induce partial differentiation in colonies of WEHI-3B myeloid leukemic cells. Recombinant GM-CSF competed effectively for the binding of 125I-labeled native GM-CSF to hemopoietic cells, and antiserum to recombinant GM-CSF also neutralized the biological activity of native GM-CSF. The bacterially synthesized GM-CSF was a slightly more effective stimulus for megakaryocyte colony formation than the native molecule. The demonstration that purified bacterially synthesized GM-CSF is biologically active in vitro now permits studies to be undertaken on the in vivo effects of this material.     

Concentration of phenylbutazone (PBZ) and 13, 14-dihydro-15 keto PGF(2alpha) (PGFM) in blood and seminal plasma of stallions treated with PBZ

Three stallions, 3 to 5 yr old and approximately 550 kg bodyweight, were used in a switchback experimental design to study the effect of daily, oral administration of 3g PBZ on the concentrations of PBZ and PGFM in blood (plasma) and seminal plasma (SP). Control and treatment periods were each 24 days' duration. Blood and semen samples were simultaneously collected every three days during these periods. Each stallion served consecutively as a control, treated, control, and treated subject for 24 days in each of the four periods. Concentrations of PBZ were obtained using HPLC and PGFM by specific RIA. Concentrations (mean +/- SE) of PBZ averaged 9.2 +/- 0.12 ug/ml in plasma but were undetectable in SP following the treatments. There was no significant difference in the plasma levels of PGFM between pre- and post- treatment values. However, there was a significant difference (P<0.001) in PGFM concentrations of seminal plasma before and after treatment. Results of this study suggest that daily, oral adminisration of 3g PBZ for 24 days to mature stallions can significantly decrease seminal plasma concentrations of PGFM. The physiological significance of this observation remains speculative.