Estrogen induction of progestophilins in rat estrogen-sensitive cells grown in media supplemented with sera from castrated rats and from rats bearing an alpha-fetoprotein-secreting hepatoma

The purpose of this work was to study the effect of alpha-fetoprotein (AFP) over cell multiplication and the induction of an estradiol-17 beta (E2)-dependent marker, i.e., progestophilins in E-sensitive cells C2(9)RAP derived from a W/Fu rat pituitary tumor. These cells proliferate in isogeneic hosts under the influence of E2, while they proliferate in culture regardless of the presence of E2. C2(9)RAP cells were grown in medium supplemented with 10% horse serum. Progestophilin levels were measured 48 h after adding serum (20% horse, or castrated rat, or AFP-secreting tumor-bearing rat) and estrogen to the 10% horse serum-supplemented medium in which the cells were growing. Maximal induction of progestophilins was obtained at 3 X 10(-10) M E2 in cells grown in medium containing horse or castrated rat serum. In contrast, maximal induction of progestophilins required 3 X 10(-8) M E2 in cells grown in medium supplemented with the serum of Morris hepatoma 7777-bearing rats. This serum contained AFP levels comparable to those present at birth in the rat. 11-Methoxy-17 beta ethynylestradiol (R2858), a synthetic estrogen with little affinity for AFP, was also tested for its ability to induce progestophilins. The degree of maximal induction of progestophilins expressed as percentage of the respective control, was similar for all experimental groups, both with E2 and with R2858. In addition, we compared the free E2 levels in the culture medium with the progestophilin levels and the cell proliferation rate. We found that the progestophilin levels were maximal at free E2 concentrations above 11 pg E2/ml, whereas there was no correlation between the free E2 levels and the proliferation rate. Moreover, the proliferation rate of cells in medium supplemented with horse or castrated rat serum was maximal at concentrations of free E2 below 0.4 pg/ml; whereas cell proliferation was inhibited with hepatoma serum even at concentrations of free E2 of 44 pg/ml. We conclude that the effect of hepatoma serum on the E2 induction of progestophilins seems to be mediated by the effect of AFP on the availability of free estrogen, since it is abolished by the addition of both natural and synthetic estrogens. The inhibitory effect of hepatoma serum upon cell proliferation is not reversed by estrogens and thus seems to be mediated by mechanisms other than E2 trapping by AFP.     

Exploring the binding pocket of quinone/inhibitors in mitochondrial respiratory complex I by chemical biology approaches

ABSTRACT

NADH-quinone oxidoreductase (respiratory complex I) is a key player in mitochondrial energy metabolism. The enzyme couples electron transfer from NADH to quinone with the translocation of protons across the membrane, providing a major proton-motive force that drives ATP synthesis. Recently, X-ray crystallography and cryo-electron microscopy provided further insights into the structure and functions of the enzyme. However, little is known about the mechanism of quinone reduction, which is a crucial step in the energy coupling process. A variety of complex I inhibitors targeting the quinone-binding site have been indispensable tools for mechanistic studies on the enzyme. Using biorationally designed inhibitor probes, the author has accumulated a large amount of experimental data characterizing the actions of complex I inhibitors. On the basis of comprehensive interpretations of the data, the author reviews the structural features of the binding pocket of quinone/inhibitors in bovine mitochondrial complex I.     

In vitro and in vivo binding of toremifene and its metabolites in rat uterus

The in vitro binding affinities of toremifene (TOR), 4-hydroxy toremifene (4-OH-TOR) and several other metabolites for the rat uterine cytosolic estrogen receptor were compared with those of tamoxifen (TAM) and 4-hydroxy tamoxifen (4-OH-TAM). Only small differences were observed and the binding affinities of both 4-hydroxy metabolites were similar to that of estradiol (E2). Uterine uptake and subcellular distribution of [3H]TOR and [3H]TAM were then compared at 1, 8 and 72 h after administration to castrated rats. The uptake and retention of both antiestrogens were similar at all times. In each case the amount of nuclear bound radioactivity declined to low levels at 8 and 72 h but the ratios of 4-OH-TAM/TAM and 4-OH-TOR/TOR determined by HPLC analysis increased dramatically at 72 h. The level of radioactivity in both plasma and uterine cytosol at 72 h was significantly higher following [3H]TAM administration. However, most of the radioactivity appeared to be in a conjugated form since it was not extractable with solvent. Finally, the ability of prior administration of each antiestrogen (100 mg/kg) to block uterine [3H]estradiol uptake was examined at 3 and 7 days. It was found that uterine wet weights were higher than control one week after administration of both compounds. Prior administration of TOR increased nuclear uptake of [3H]E2 whereas TAM had no effect. The results of these experiments suggest that toremifene and tamoxifen have very similar in vitro and in vivo binding properties but differences in metabolism exist that may be important.     

Dispersal and reproductive properties of sugarcane bugs,Cavelerius saccharivorus

The distribution of sugarcane bugs, Cavelerius saccharivorus is remarkably patchy at time of the peak emergence of the first generation adult in early summer. In population near carrying capacity 70% of adults become macropterous, and they move to sparse populations or vacuum areas. The experiment on the reproductive difference between females from field populations with different densities, showed that the overcrowding restricted their oviposition. On the further experiment it was shown that reproductive ability of macropterous adults or dispersers was not much inferior to that of brachypterous from sparse populations. Overcrowding at time of the peak emergence is reduced by the dispersal. Hence the intraspecific competition is a transient condition. The oviposition is at first restricted in residents of dense populations as compared with brachypterous adults of sparse populations, but the difference becomes small as the former density decreases by the emigration. The last section considers the relation of the reproductive rate and dispersal risk to the rate of dispersal with the simple model, which explains the characteristic dispersal of this species.     

alpha-Lactalbumin: a calcium metalloprotein

Metal analyses and the studies of the effects of EDTA on unfolding reactions have shown that α-lactalbumin is a calcium metalloprotein. The role of the calcium binding in its biological activity is considered. A plausible site of binding is presented on the basis of the metal-binding site of lysozyme and of the structural models of the protein based on the lysozyme structure.     

Characterization of soluble lignin released from alfalfa by sheep digestion

A faecal soluble lignin fraction (FSL) extracted with 90% dioxane from the faeces of sheep fed on alfalfa hay was characterized by chemical analysis, nitrobenzene oxidation, fourier transform infrared spectroscopy and gel permeation chromatography, and compared with milled wood lignin (MWL) isolated from the alfalfa hay. The amount of FSL in the faeces was low, accounting for only 1% of the lignin present in the alfalfa hay. FSL and MWL consisted mostly of lignin components and contained a small amount of carbohydrate. FSL had a much higher proportion of syringylpropane units than MWL and showed a wide molecular size distribution. The results indicate the selective and limited solubilization of syringyl-rich lignin from alfalfa by sheep digestion.     

Fear responses of Japanese monkeys to scale models

This study investigated fear responses of Japanese monkeys (Macaca fuscata) to scale models. Fear responses of participants were assessed using rating scores assigned by judges. In Experiment 1, participants were presented with scale models of objects in furniture, vehicle, and mammal categories. Overall, the participants expressed stronger fear responses to mammal objects as compared to the other two kinds. In Experiment 2, participants were presented with scale models of furniture, new mammals, and animals composed of insects, birds, fish, and reptiles. As a whole, the participants showed stronger fear responses to the new mammal and animal objects than to furniture objects. Fear responses to mammal and animal objects were comparable. These results suggest that Japanese monkeys show stronger fear responses to objects possessing perceptual properties of animals, as opposed to objects that do not possess such properties.