The effect of prostaglandins A2,E1,E2,15 methyl E2, 16, 16 dimethyl E2 and F2α on erythropoiesis

Prostaglandins A₂, E₁, E₂, methylated E₂s and F₂α affected erythropoiesis and/or erythropoietin (Ep) production. This action is indicated in the exhypoxic, polycythemic mouse where radioiron incorporations into RBC increased after administration of these compounds. The kidney and liver have been indicated through previous studies, to actively participate in Ep production. By the removal of one of these active sites in a murine system treated with prostaglandins it is shown that a response is reflected in Ep levels. Interference of the action of prostaglandins (PG) is altered by the removal of one of these target sites of Ep production. The erythropoietic responses elicited by PGA₂, E₁, and perhaps the methylated PGE₂s act through the liver whereas PGE₂ may operate through a renal pathway for its response. PGF_2α reveals no effect on erythropoietic activity and is no different than that observed for vehicle-treated controls. The prostaglandins tested appear to act primarily through the kidney or liver but the possibility exists that some yet undetermined organ site may also be involved.     

Carnitine uptake and fatty acid utilization by diploid cells aging in culture

Uptake of carnitine by cultured human fetal lung flbroblasts (WI-38 and IMR-90) and by smooth muscle cells from calf aorta and from human uterus was found to be temperature dependent and saturable. IMR-90 cells showed an apparent K_m of 6–8 μM and a V of 21–28 pmol/h/10⁶ cells for l-carnitine. Transport was abolished by N-ethylmaleimide and was inhibited variably by octanoyl-d-carnitine, d-carnitine, and carbonyl cyanide p-trifluoromethoxyphenylhydrazone. Although WI-38 and IMR-90 cells accumulate lipids as they age in culture, they take up carnitine as rapidly as do smooth muscle cells of aorta and uterus that do not exhibit such accumulation. Comparison of the rates of carnitine uptake by IMR-90 fibroblasts during the logarithmic phase of growth shows no difference between “young” and “old” cultures. In contrast, when confluent or postconfluent monolayers were compared and uptake expressed as a function of cell number, cells grown from late passages took up carnitine more rapidly than did cells grown from early passages. However, when account was taken of cell size, and carnitine expressed as a function of cell volume, the differences in carnitine uptake between early and late passages were no longer apparent for the confluent or postconfluent monolayers examined. Moreover, late passage fibroblasts took up and oxidized radioactive palmitate at least as rapidly as did cells from early passages. Our results suggest that accumulation of lipid in aging fibroblasts is not due to decreased carnitine uptake or fatty acid oxidation.     

Comutagenesis of sodium arsenite with ultraviolet radiation in Chinese hamster V79 cells

Summary Solar ultraviolet radiation has been associated with the induction of skin cancer. Recent studies have indicated that near-ultraviolet, especially UVB, is mutagenic. Exposure to trivalent inorganic arsenic compounds has also been associated with increased skin cancer prevalence. Trivalent arsenic compounds are not mutagenicper se, but are comutagenic with a number of cancer agents. Here, we test the hypothesis that arsenite enhances skin cancer via its comutagenic action with solar ultraviolet radiation. Irradiation of Chinese hamster V79 cells with UVA (360 nm), UVB (310 nm) and UVC (254 nm) caused a fluence-dependent increase in mutations at thehprt locus. On an energy basis, UVC was the most mutagenic and UVA the least. However, when expressed as a function of toxicity, UVB was more mutagenic than UVC. Nontoxic concentrations of arsenite increased the toxicity of UVA, UVB and UVC. Arsenite acted as a comutagen at the three wavelengths; however, higher concentrations of arsenite were required to produce a significant (P < 0.05) comutagenic response with UVB. The increased mutagenicity of UVB and UVA by arsenite may play a role in arsenite-related skin cancers.     

Expression of glucoamylase gene usingSUC2 promoter inSaccharomyces cerevisiae

Summary The cloning of glucoamylase geneSTA using theSUC2 promoter intoSaccharomyces cerevisiae was performed. The signal sequence ofSTA gene was used for the secretion of glucoamylase protein. The plasmid constructed in this way was named YEpSUCSTA and its expression was identified. The expression of YEpSUCSTA was repressed in the presence of glucose in growth medium, but derepressed when glucose became depleted. YEpSUCSTA showed the similar efficiency of glucoamylase secretion as YEpSTA-F which has the entireSTA gene. Glucoamylase activity in starch-glucose medium was largely increased because cell mass and plasmid stability were high in biosynthesis phase compared to extracellular glucoamylase activities in media which starch or glucose was the only carbon source.     

Induced feeding preferences in dicofol-resistant two-spotted spider mites (Acari: Tetranychidae)

Preadult rearing conditions affected the behavior of dicofol-resistant two-spotted spider mites (Tetranychus urticae). Resistant spider mites reared on dicofol-treated leaves initiated a significantly greater number of feeding bouts on dicofol-treated leaves than did genetically identical spider mites reared on residue-free leaves. Therefore the prior exposure of resistant spider mites resulted in induced feeding preferences that could exacerbate the potential outcome of the resistance by resulting in greater amounts of feeding by resistant individuals on dicofol-treated areas. Since resistant individuals that had not experienced dicofol in their lifetime did not display this feeding preference, avoidance of this phenomenon of induced feeding preference may be an undescribed value of rotations of pesticides.     

Thermal stability of turnip yellow mosaic virus RNA: effect of pH and multivalent cations on RNA deaggregation and degradation.

Light scattering studies of RNA isolated from turnip yellow mosaic virus (TYMV) revealed a molar mass of 1.9.10(6) g mol-1, which is close to the value of 2.0.10(6) g mol-1 published for intact genomic TYMV RNA (2M RNA). However, gel electrophoresis under denaturing conditions demonstrated that only 30-40% of this native RNA was 2M RNA. Sucrose gradient centrifugation revealed the occurrence of a series of smaller RNA size classes, the mass ratios of which were greatly influenced by the pH of the solution and the presence of EDTA. These results suggest that native TYMV RNA preparations originally contain a mixture of intact RNA particles and of aggregates of RNA fragments with the same molar mass of about 2.10(6) g mol-1, and that the size classes are intermediates in the deaggregation process of the degraded genomic TYMV RNA. The native RNA displayed pH-dependent deaggregation and degradation. The degradation process of 2M RNA followed (pseudo) first-order kinetics. Lower degradation rates were observed for RNA depleted of divalent cations and polyamines. For depleted 2M RNA an enthalpy of activation of about 100 kJ mol-1 and an almost zero entropy of activation was calculated. Similar values were also found for depleted E. coli ribosomal RNAs and depleted MS2 RNA, demonstrating that all RNAs are equally vulnerable to degradation. In the presence of multivalent cations the activation enthalpy for 2M TYMV RNA degradation increased to 150 kJ mol-1 and the entropy of activation to 150 J K-1 mol-1, indicative for a different degradation mechanism.     

Functional anion binding sites in dogfish M4 lactate dehydrogenase

X-ray diffraction data have been collected from dogfish M₄ lactate dehydrogenase crystals in which ammonium sulfate had been exchanged by citrate at pH 6.0 and 7.8. Data were also collected from crystals which had been soaked in 0.1 m oxamate, a lactate dehydrogenase inhibitor. The difference electron density maps obtained have been interpreted in terms of two exchangeable anion binding sites, one at the active center and one between two subunits. The active center site is coincident with the substrate binding site in a ternary complex, while the subunit boundary site, which has been observed in several different forms of the enzyme, may be involved in stabilizing the tetramer.     

Can we rely on forest reserves for primate conservation?

Tropical forests contain much of the world's biodiversity, yet their rate of decline is increasing. The strategy most frequently used to protect this biodiversity is to make parks and reserves. While there is a great deal of research on the effectiveness of parks for protecting biodiversity, there is little research on how well extractive reserves conserve biodiversity. Here, we evaluate the effectiveness of four forest reserves in western Uganda at maintaining populations of primates and compare census data from the reserves to data from the neighbouring well‐protected Kibale National Park. The relative abundance of the five most common primates in the park was approximately four times that of the forest reserves. In the forest reserves, evidence of new human encroachment was seen every 500 m, while in the park it was seen every 100,000 m. Two recommendations emerge from our research: (i) for forest reserves, such as those studied here, to have conservation value for primates, extraction must be reduced and (ii) until the long‐term viability of the populations in forest reserves can be ascertained, they should not be considered in estimates of the sizes of endangered species protected ranges.     

Cold Shock Domain Containing E1 (CSDE1) Protein is Overexpressed and Can be Targeted to Inhibit Invasiveness in Pancreatic Cancer Cells

Pancreatic cancer has a dismal prognosis and to date there are no targeted therapies for this malignancy. Using shotgun proteomics, the mRNA binding protein cold shock domain containing E1 (CSDE1), also called upstream‐of‐N‐Ras, is detected in pancreatic cancer cell lines but not in normal pancreatic epithelial cells. The expression of CSDE1 in pancreatic cancer cells is confirmed by Western blotting and immunohistochemistry of human pancreatic tumors. In vitro functional assays show that siRNA downregulation of CSDE1 or gene knockout using CRISPR‐Cas9 significantly reduce the invasiveness of pancreatic cancer cells. Together, this study reveals that CSDE1 is overexpressed in pancreatic cancer and is a potential therapeutic target to inhibit pancreatic cancer cell invasion.