Interaction of chicken gizzard smooth muscle calponin with brain microtubules.

Calponin, a major actin-, tropomyosin-, and calmodulin-binding protein in smooth muscle, interacted with tubulin, a main constituent of microtubules, in a concentration-dependent fashion in vitro. The apparent K(d) value of calponin to tubulin was calculated to be 5.2 microM with 2 mol of calponin maximally bound per 1 mol of tubulin. At low ionic strength, tubulin bound to calponin immobilized on Sepharose 4B, and the bound protein was released at about 270 mM NaCl. Chemical cross-linking experiments showed that a 1:1 molar covalent complex of calponin and tubulin was produced. The amount of calponin bound to microtubules decreased with increasing ionic strength or Ca2+ concentration. The addition of calmodulin or S100 to the mixture of calponin and microtubule proteins caused the removal of calponin from microtubules in the presence of Ca2+, but not in the presence of EGTA. Calponin-related proteins including tropomyosin, SM22, and caldesmon had little effect on the calponin binding to microtubules, whereas MAP2 inhibited the binding. Interestingly, there was little, if any, effect of mycalolide B-treated actin on the binding of calponin to microtubules. Furthermore, only about 20% of calponin-F-actin interaction was inhibited in the presence of an excess amount of tubulin (4 mol per mol of calponin), indicating that tubulin binds to calponin at a different site from that of actin. Compared with MAP2, calponin had little effect on microtubule polymerization.

Studies on the therapeutic effect of 2-pyridine aldoxime methiodide (2-PAM) in mammals following organophosphorus compound-poisoning (report III): distribution and antidotal effect of 2-PAM in rats.

The metabolic fate of 2-PAM and its antidotal effect on organophosphorus compound poisoning in rats were studied. When 14C-2-PAM was administered intravenously, the amount of 14C reaching the brain was small. Following administration by intramedullary injection, 14C was present in high concentrations in the brain, and 72-90% of the 14C present in the brain corresponded to the unchanged form of 2-PAM. 2-PAM was rapidly excreted into the urine and feces following either intramedullary or intravenous administration. The half-life of 2-PAM in the brain following intramedullary administration was 1.52 hr. Intramedullary administration of 2-PAM to rats poisoned with fenitrothion or malathion enabled their survival and induced reactivation of brain cholinesterase.

Studies on the therapeutic effect of 2-pyridine aldoxime methiodide (2-PAM) in mammals following organophosphorus compound (OP)-poisoning (report II): aging of OP-inhibited mammalian cholinesterase.

We studied the ability of 2-PAM to reactivate cholinesterase (ChE) inhibited by organophosphorus compounds (OPs) and aging. We estimated the reactivation rate with 2-PAM following inhibition of ChE by fenitrothion, methylparathion or ethylparathion using erythrocytes of rat and rabbit and rat brain. The period of time during which inhibited ChE could be reactivated was shorter in the case of inhibition by fenitrothion or methylparathion than in the case of inhibition by ethylparathion. This results suggest that aging is related to the presence of the alkyl group in OPs, and occurs faster in the case of inhibition by OPs with an O,O-dimethyl moiety than in the case of inhibition by OPs with an O,O-diethyl moiety.

Pyrethroids and the striatal dopaminergic system in vivo.

1. Type I (permethrin and allethrin) or type II (cypermethrin and fenvalerate) pyrethroids caused 23-37% increases in the striatal content of the dopamine metabolite 3,4-dihydroxyphenylacetic acid (DOPAC). 2. Toxicity symptoms and increases in DOPAC were associated with higher brain concentrations for type I (2.6-5.8 micrograms/gm) than type II pyrethroids (0.4-0.6 micrograms/gm). 3. No specific difference in the interaction between type I and II pyrethroids and the striatal dopaminergic system were recognized.

Fenvalerate-induced granulomatous changes in rats and mice.

Wistar rats and ddY mice were fed diet containing 10 to 3000 ppm technical fenvalerate for the major portion of their life span. Microgranulomatous changes were observed in lymph nodes, liver and spleen of both rats and mice as well as in adrenals of rats. The no-effect level for microgranulomatous changes was 30 ppm for mice and 150 ppm for rats. Transmission electron microscopy of phagocytes in the granulomatous foci revealed needle- or rod-shaped crystalline bodies within the cytoplasm. To examine reversibility of the microgranulomatous changes, ddY mice were fed the diet containing technical fenvalerate at the levels of 1000 and 3000 ppm for 6 weeks, followed by rearing on the control diet for up to 12 months. The size and number of the microgranulomatous changes were reduced with time. The microgranulomatous changes were typical of foreign body granulomas; they did not have the appearance of granulomas formed in response to immunologic stimulus.