Potentiation of the effects of bradykinin on its receptor in the isolated guinea pig ileum

Angiotensin I-converting enzyme (ACE/kininase II) inhibitors potentiated guinea pig ileum's isotonic contractions to bradykinin (BK) and its analogues, shifting the BK dose-response curve to the left. ACE inhibitors added at the peak of the contraction immediately enhanced it further (343 +/- 40%), although the ileum inactivated BK slowly (t(1/2) = 12-16 min). Chymotrypsin and cathepsin G also augmented the activity of BK up to three- or four-fold, but in a manner slower than that of ACE inhibitors. The BK B(2) receptor blocker HOE 140 inhibited all effects. Histamine and angiotensin II were not potentiated. ACE inhibitors potentiate BK independent of blocking its inactivation by inducing crosstalk between ACE and the BK B(2) receptor; proteases activate the receptor by different mechanism.     

DNA methylation patterns in tissues from mid-gestation bovine foetuses produced by somatic cell nuclear transfer show subtle abnormalities in nuclear reprogramming

Background  

Cloning of cattle by somatic cell nuclear transfer (SCNT) is associated with a high incidence of pregnancy failure characterized by abnormal placental and foetal development. These abnormalities are thought to be due, in part, to incomplete re-setting of the epigenetic state of DNA in the donor somatic cell nucleus to a state that is capable of driving embryonic and foetal development to completion. Here, we tested the hypothesis that DNA methylation patterns were not appropriately established during nuclear reprogramming following SCNT. A panel of imprinted, non-imprinted genes and satellite repeat sequences was examined in tissues collected from viable and failing mid-gestation SCNT foetuses and compared with similar tissues from gestation-matched normal foetuses generated by artificial insemination (AI).     

Reduction of adenine nucleotide content of clone 4 MDCK cells: effects on multiplication, protein synthesis, and morphology

The antitumor agent hadacidin (N-formyl-hydroxyamino-acetic acid), at 4 mM, inhibited the multiplication of clone 4 Madin Darby canine kidney (MDCK) cells within 24 hr. Growth resumed rapidly upon replacement of hadacidin with aspartate, an observation consistent with the drug's action as a competitive inhibitor of adenylosuccinate synthetase, an enzyme in adenine nucleotide biosynthesis. Data indicate that the drug-treated cells were arrested in S phase of the cell cycle. Accompanying inhibition of multiplication was a 16-fold increase in the area occupied by the cells and a refractoriness to release by treatment with trypsin. None of these changes occurred when 0.5 mM adenosine was included in the incubation mixture containing the inhibitor. Hadacidin decreased the adenosine triphosphate (ATP) and cyclic adenosine monophosphate (cAMP) content of the cells as well as the rate at which 3H-leucine was incorporated into protein. In the presence of 1 mM dibutyryl cAMP and theophylline, the drug had no effect on cell division and protein synthesis. The data suggest that, in clone 4 MDCK cells, the effects of hadacidin are mediated by diminishing the level of cAMP.     

Morphologic and functional correlates of plasma membrane injury during oxidant exposure

Plasma membrane injury by exposure to hydrogen peroxide was examined in a renal epithelial cell line (LLC-PK1). Morphologic and functional parameters of plasma membrane integrity were studied in an attempt to eludicate the sequence of membrane alterations during the evolution of hydrogen peroxide-mediated injury. These parameters included plasma membrane potential and permeability, plasma membrane bleb formation, cellular size, and plating efficiency. Plasma membrane potential was the earliest parameter affected by hydrogen peroxide exposure. Half maximal depolarization occurred within 15-30 min of exposure to 1 mM, after 10-15 min exposure to 100 mM and after over 150 min exposure to 10 microM hydrogen peroxide. After exposure to 1 mM hydrogen peroxide, the following sequence of events was seen; increased plasma membrane blebbing (30 min), cell swelling (90-125 min) and increased plasma membrane permeability (150-240 min). After a 30 min exposure to 1 mM hydrogen peroxide, cellular plating efficiency, measured at 24 h, was reduced by 50% (P less than .001). These changes were accelerated, although their order of appearance was unchanged, at higher concentrations of hydrogen peroxide. We conclude that functional and morphologic expressions of cellular injury in this model occur in a defined sequence with plasma membrane depolarization representing the earliest marker of membrane injury during hydrogen peroxide exposure.     

Water and electrolyte balance in protoscoleces of Echinococcus granulosus incubated in vitro: Effect of metabolic inhibitors

Reisin I.L., Rabito C.A. and Cantiello H.F. 1981. Water and electrolyte balance in protoscoleces of Echinococcus granulosus incubated in vitro: effect of metabolic inhibitors. International Journal for Parasitology 11: 405–410. The effects of metabolic inhibitors on the Na⁺, K⁺, Cl^? and water balance of protoscoleces of Echinococcus granulosus (sheep strain) were studied in vitro. The protoscoleces were incubated at 37°C in Ringer Krebs solution for 3 h in the presence of iodoacetate, 3 mM (IA); potassium cyanide, 3 mm (KCN); 2?4 dinitrophenol, 0.2 mm (DNP); ouabain, 10^?M or ethacrynic acid 0.5 mm. The effects of IA and/or KCN on the water and electrolyte balance were tested at high (0.95 × 10⁵Pa) and low (0.05 × 10⁵ Pa) oxygen tensions. Inhibitors produced a decrease in K⁺ as well as an increase in Na⁺ contents. At both high and low O₂ tensions the Na⁺ balance was greatly altered by IA, the action of which could be already observed during the first hour of treatment. The cations did not reach a steady state balance during 3 h of incubation. At high oxygen tension Na⁺ and K⁺ balance was also altered by KCN or DNP though their actions were not as marked as that of IA. Ouabain affected the Na⁺ and K⁺ contents that reached new steady state distribution between 1.5 and 3 h of treatment while water and electrolyte contents were not modified by ethacrynic acid. In all the experiments no changes in Cl^? and water contents were observed. It is concluded that the energy required to maintain the Na?K balance mechanisms within protoscoleces is largely provided by the anaerobic glycolytic pathway and that the aerobic oxidative pathway contribution to the energy balance is only accessory.