A microspectrofluorometric approach for the study of Benzo(a)pyrene and Dibenzo(a,h)anthracene metabolization in single living cells

Summary The use of a microspectrofluorometer in conjunction with the microelectrophoretic intracellular injection of glycolytic intermediates, for the study of carcinogen metabolization (i.e. Benzo(a)pyrene (BP) and Dibenzo(a, h)anthracene) by single living EL2 ascites cells, has allowed the detection of fluorescence attributable to the metabolites of BP and DBA.The results obtained are in agreement with those yielded by more conventional methods: i.e. in vivo assay and in vitro reconstitution of intracellular environment. Thus, it is observed that NADPH is more active in the metabolization of BP. Furthermore, in the case of BP the fluorescence attributable to a BP metabolite, exhibits a strong similarity to 3-OH benzo(a) pyrene.     

The cytochromes of the filamentous bacteriaStreptomyces clavuligerus andSaccharopolyspora erythraea (FormerlyStreptomyces erythraeus)

The cytochromes of the filamentous bacteriaStreptomyces clavuligerus andSaccharopolyspora erythraea have been studied by low-temperature (77K) difference spectra and room-temperature potentiometric titrations. Difference spectra of membranes fromSa. erythraea indicate the presence of twoc- and twob-type cytochromes. A furtherb-type cytochrome is revealed by potentiometric titration. The soluble and membrane-bound forms of a green pigment, believed to be involved in electron transport to oxygen, are each shown to have complex absorption spectra which, in the case of the soluble form, may be potentiometrically resolved into several components. Threeb-type and threec-type cytochromes are shown to be present in the membranes ofSt. clavuligerus.     

Cyclic AMP decreases LDL catabolism and cholesterol synthesis in the human hepatoma cell line HepG2

A 24h pretreatment of the human hepatoma cell line HepG2 with dibutyryl cyclic AMP in the presence of theophylline induced a dose dependent decrease in low density lipoprotein binding, uptake and degradation. This effect is most likely due to a reduction of the LDL receptor number. Sterol synthesis from sodium acetate is markedly inhibited, either in the presence or absence of LDL, whereas synthesis from mevalonic acid is unchanged. Cyclic AMP also induced a decrease in hydroxy methyl glutaryl coenzyme A reductase activity. These effects of cyclic AMP might be involved in some hormonal regulation of the LDL pathway and cholesterol metabolism in the liver.     

Use of an anti-B monoclonal antibody of mouse origin with the Groupamatic (MG 50) apparatus

This study refers to the use of a monoclonal anti-B antibody of murine origin on Groupamatic MG 50. The monoclonal antibody produces results similar to those obtained with human immune antibodies.     

Age-associated adipose tissue inflammation promotes monocyte chemotaxis and enhances atherosclerosis

Although aging enhances atherosclerosis, we do not know if this occurs via alterations in circulating immune cells, lipid metabolism, vasculature, or adipose tissue. Here, we examined whether aging exerts a direct pro-atherogenic effect on adipose tissue in mice. After demonstrating that aging augmented the inflammatory profile of visceral but not subcutaneous adipose tissue, we transplanted visceral fat from young or aged mice onto the right carotid artery of Ldlr^−/− recipients. Aged fat transplants not only increased atherosclerotic plaque size with increased macrophage numbers in the adjacent carotid artery, but also in distal vascular territories, indicating that aging of the adipose tissue enhances atherosclerosis via secreted factors. By depleting macrophages from the visceral fat, we identified that adipose tissue macrophages are major contributors of the secreted factors. To identify these inflammatory factors, we found that aged fat transplants secreted increased levels of the inflammatory mediators TNFα, CXCL2, and CCL2, which synergized to promote monocyte chemotaxis. Importantly, the combined blockade of these inflammatory mediators impeded the ability of aged fat transplants to enhance atherosclerosis. In conclusion, our study reveals that aging enhances atherosclerosis via increased inflammation of visceral fat. Our study suggests that future therapies targeting the visceral fat may reduce atherosclerosis disease burden in the expanding older population.     

1H, 15N and 13C assignments of a putative peptidyl prolyl cis–trans isomerase FKBP12 from Trypanosoma brucei

TbFKBP12 is a putative peptidyl prolyl cis–trans isomerase from Trypanosoma brucei, causative agent of the African trypanosomiasis or sleeping sickness. It interacts with the immunosuppressive drug rapamycin inhibiting the formation of TORC2 complex leading to parasite death by inhibiting cell proliferation through cytokinesis blockade. Moreover, RNAi silencing of TbFKBP12 revealed essential function in both procyclic and bloodstream forms. Both facts make TbFKBP12 an attractive target for ligand development and thus structural data is desirable. In this work we report the NMR resonance assignments for ¹H, ¹⁵N and ¹³C nuclei in the backbone and side chains of the TbFKBP12 as basis for further studies of structure, backbone dynamics, interaction mapping and drug screening.     

Disappearance of prostaglandins F2alpha and 15-methyl F2alpha from amniotic fluid as measured by radiommunoassay.

A sensitive and relatively specific radioimmunoassay for 15 (S) 15 methyl prostaglandin F2alpha was used to determine the levels of the drug in amniotic fluid after it had been injected intra-amniotically for termination of second trimester pregnancy. The disappearance of the free acid (tham salt) and methyl ester of the prostaglandin analogue were similar. The results of this preliminary study suggest that the drug rapidly equilibrates in the fluid and this followed by a slow removal from the amniotic sac. A comparison with a similar study with PGF2Alpha revealed that the analogue had a longer half-life in the amniotic fluid.