Effects of U-40481 and formetanate on the isolated rabbit central ear artery

Formetanate, a formamidine-type pesticide, and U-40481 (N-methyl-N′-2,4-xylylformamidine), a metabolite of amitraz, also a formamidine pesticide, contract the rabbit central ear artery with their maximal contractions being 22 ± 8% and 49 ± 6% of norepinephrine contractions, respectively. Maximal contractions were obtained with 10^?3M formetanate and 10^?4M U-40481, and cumulatively added higher concentrations caused a decrease in tension from that maximum. Their contractions were antagonized by 10^?6M and 3 × 10^?6M phentolamine. U-40481 reversibly antagonized contractions induced by serotonin, norepinephrine, and histamine, and to some extent potassium. Formetanate had little antagonist activity. Neither compound altered the resting rate of washout of radioactivity from [³H]norepinephrine preloaded strips. Both reduced electrically induced release, which may be related to local anesthetic-like actions on sympathetic neurones. Thus both compounds are partial agonists at the α-adrenergic receptor, and reduce electrically induced norepinephrine release, and U-40481 antagonizes contractions induced by certain other vasoactive agents.     

Studies on the action of chlordimeform in cockroaches

The effects of chlordimeform [N-(2-methyl-4-chlorophenyl)-N′,N′-dimethylformamidine] on amine regulatory mechanisms in insects were studied. Chlordimeform inhibits monoamine oxidase (MAO) from cockroach heads in vitro, and the MAO substrates serotonin and norepinephrine accumulate in poisoned insects in vivo. Chlordimeform synergizes the toxicity of tryptamine, another MAO substrate. The significance of these findings in relation to the mode of action of chlordimeform is discussed.     

Inhibition of oviposition in the cattle tick Boophilus microplus by certain acaricides

Doses of ten acaricides, ranging from 2.5 to 10 μg, induced about 10-60% mortality and inhibited oviposition in engorged Boophilus microplus as well as preventing larval hatching from the eggs. The effects of the acaricides were dose dependent. The efficacy of acaricides in reducing the reproductive potential (inhibition of oviposition × inhibition of hatching/100) was in the following order: dimethoate > dioxathion > naled > diazinon > chlordimeform > carbaryl > trichlorphon > phosphamidon > gamma-HCH > fenitrothion. Most of the acaricides also retarded the oviposition cycle, delaying the peak activity by 4-8 days. The ovarian development in control ticks reached its peak between the 5th and 7th days before decreasing gradually and ceasing completely between the 16-20th days after engorgement. The protein content of the ovaries and the rate of incorporation of [¹⁴C]glycine into the tissues followed a similar pattern. Carbaryl, fenitrothion and naled inhibited these activities. Chlordimeform stimulated [¹⁴C]glycine incorporation and increased the ovarian protein content up to the 13th day before resorption of the oocytes. The eggs from treated ticks were mostly non-viable and contained more protein; those from dioxathion, carbaryl and chlordimeform treatments had a higher dry weight than the control.     

Effects of chlordimeform on rectus abdominis muscle of frog

The effects of chlordimeform on rectus abdominis muscle of frog were investigated. Chlordimeform (10^?3M) caused a slow contraction, and at lower concentration (10^?5–10^?3M) it inhibited the acetylcholine-induced contraction in noncompetitive manner. When chlordimeform was applied to the muscle of Rana catesbiana, K⁺-induced contraction was also inhibited in noncompetitive manner. Whereas it had no effect on caffeine-induced contraction.Chlordimeform-induced contraction was not affected by respective addition of d-tubocurarine (10^?4M), procaine (10^?3M), or eserine (0.3 mM), which results were same as that of K⁺-induced contraction. Chlordimeform, at lower concentration (10^?5–10^?3M), inhibits the acetylcholine- and K⁺-induced contractions probably owing to depression of not only the sensitivity of endplate but also the excitability of cell membrane.     

Mechanism of neuromuscular block by chlordimeform

The mechanism of action of chlordimeform on the frog sciatic nerve-sartorius muscle preparation has been studied by means of microelectrodes. Chlordimeform (0.1 mM) suppresses the amplitude of spontaneous miniature end-plate potentials without much changing their frequency. At 1 mM, it completely blocks the miniature end-plate potentials. The resting membrane potential is not affected. The end-plate potential evoked by nerve stimulation is also suppressed, while the action potential from the nerve terminal is not impaired. The sensitivity of the end-plate membrane to the transmitter substance as examined by iontophoretic applications of acetylcholine is effectively decreased by chlordimeform. The quantal content of transmitter release is not affected. It is concluded that the block of neuromuscular transmission by chlordimeform is due primarily to a depression of the end-plate sensitivity to the transmitter.     

Actions of formamidines,local anesthetics,octopamine and related compounds upon the electrical activity of neurohaemal organs of the stick insect (Carausius morosus) and sense organs of fly larvae (Musca domestica; Calliphora erythrocephala)

The pesticides, chlordimeform and amitraz, and their metabolites, demethylchlordimeform, N¹-(2,4-dimethylphenyl)-N-methylformamidine, and 2,4-dimethylformanilide, are effective at concentrations as low as 3 μM in raising the firing rate of endogenously active neurosecretory fibres in the isolated neurohaemal organs of Carausius morosus. Molecules such as bunamidine and cetrimide, with cationic detergent properties, produced sporadic bursting which did not elevate the overall firing rate to any great extent. Indeed, bunamidine could induce complete block of action potentials at concentrations as low as 30 μM. The local anesthetics, procaine, lidocaine, and benzocaine, do not induce block of activity at least up to a level of 1 mM. They have no effect at concentrations lower than 100 μM. Between 100 μM and 1 mM lidocaine and benzocaine produce a small increase in firing rate. Procaine produced a pronounced increase in the frequency of firing. The phenolic amines, octopamine, synephrine, and tyramine, markedly increased electrical activity. The catecholamines, dopamine, noradrenaline, and adrenaline, by contrast, only produced a weak excitation. Neither α- nor β-adrenergic blocking agents were effective in antagonizing the electrical activity induced by chlordimeform or phenolic amines until relatively high concentrations of about 1 mM were used. Chlordimeform was able to induce high-frequency bursts from sense organs associated with the epidermis and body-wall musculature in larvae of Musca domestica and Calliphora erythocephala. Octopamine did not induce any similar bursting activity in these sense organs. These results are discussed in relationship to current views on the mode of action of the N-aryl amidines. It is concluded that the excitatory effects of these compounds upon neurohaemal organs and sense organs are more likely to result from a direct action upon voltage sensitive channels of the nerve membranes, rather than by an effect mediated by interactions with octopamine receptors.     

Paraoxon formation and hydrolysis by mammalian liver

In vitro studies of the desulfuration of parathion at 37°C by hepatic tissue from males and females of nine mammalian species revealed sex and species variation in initial rates of parathion desulfuration and arylesterase-catalyzed hydrolysis of the oxygen analogue, paraoxon. Double reciprocal plots of initial rates of parathion activation for representative males and females of each species gave K_m values ranging from 0.2 × 10^?4?1.0 × 10^?4M parathion. Guinea pigs and rats were the only animals showing sex differences in activation, males possessing higher desulfurating abilities than the corresponding females. Based upon the sex possessing the higher desulfurating ability, the species pattern of decreasing activity was hamster > guinea pig > mouse > rat > rabbit > bovine > dog > porcine > cat. Studies of paraoxon hydrolysis indicated that only rats showed sex differences in hydrolysis, males possessing higher arylesterase activity than females. The species pattern of decreasing hydrolytic activity was in the order mouse > bovine > rat > guinea pig > rabbit > hamster > cat > dog > porcine.     

Enzymatic hydrolysis of diazoxon by rat tissue homogenates

The enzymatic hydrolysis of ³²P-labeled diazoxon was studied using tissue homogenates of rat and American cockroach. The order of the hydrolytic activities of rat tissues for diazoxon was as follows: liver > blood > lung > heart > kidney > brain. A liver enzyme hydrolyzing diazoxon to diethyl phosphoric acid and 2-isopropyl-4-methyl-6-hydroxypyrimidine was located in the microsomes. The activity of the microsomal enzyme was inhibited by EDTA, heavy and rare earth metal ions, and SH reagents. Ca²⁺ activated the enzyme and protected it from inactivation. Mitochondrial and soluble enzymes from liver and a serum enzyme also hydrolyzed diazoxon and they were also activated by Ca²⁺. The removal of calcium bound to the microsomal enzyme protein by dialysis against EDTA led to a partially irreversible change of the enzyme. The hydrolysis of diazoxon by the Ca²⁺-requiring microsomal and serum enzymes was more rapid than that of paraoxon. Hydrolysis of diazoxon did not occur in American cockroach homogenates. This difference in the capacity to hydrolyze diazoxon between mammals and insects is discussed in relation to the selective toxicity of diazinon.     

Effect of tri-o-cresyl phosphate and methamidophos on Ca uptake by brain synaptosomes in hens

The effect of tri-o-cresyl phosphate (TOCP) and methamidophos (MET) on potassium-stimulated ⁴⁵Ca uptake by brain synaptosomes in hens was studied. An in vivo test showed that TOCP increased potassium-stimulated calcium uptake 2 h after its administration, but that verapamil suppressed the enhancement of this calcium uptake. An in vitro test showed that lower concentrations of TOCP stimulated calcium uptake by synaptosomes, but that higher concentrations inhibited the uptake. In contrast, all tested concentrations of MET obviously inhibited calcium uptake; however, since calcium uptake was decreased by the administration of verapamil plus either TOCP or MET, the mechanism by which TOCP affects the voltage-operated calcium channel may be different from that of MET. The disruption of calcium homeostasis may be involved in organophosphate-induced delayed neurotoxicity (OPIDN). Calcium channel blocker may ameliorate OPIDN by maintaining calcium homeostasis in nerve cells.     

The effect of single and combined doses of chlordimeform and permethrin on cAMP and cGMP levels in the moth,Mamestra configurata Wlk

Treatment of adult males of the moth, Mamestra configurata, with chlordimeform caused large increases (up to 27-fold) in the concentration of cAMP in the head, thorax, and abdomen but had little effect on cGMP. This pesticidal effect, termed A⁺G⁰, likely results from the interaction of chlordimeform (or a metabolite) with an octopamine-sensitive adenylate cyclase present in the brain and other tissues. Treatment of moths with permethrin caused substantial increases (up to 3.4-fold) in the concentration of cGMP in the head and thorax but had little effect on cAMP. This effect, termed A⁰G⁺, may result from a disturbance of cholinergic function. Treatment of moths with a combined dose of chlordimeform and permethrin caused increases in the concentration of both cAMP and cGMP (A⁺G⁺ effect) in the head and thorax and had a synergistic effect on mortality. The synergistic effect on mortality appears to be caused by the disruption of distinct and different physiological systems by the A⁺G⁰ and A⁰G⁺ pesticides. The hypothesis that A⁺G⁰ and A⁰G⁺ pesticides, when combined, will exhibit synergism is supported by examples from the literature.     

[3H]Muscimol binding to a putative GABA receptor in honey bee brain and its interaction with avermectin B1a

A putative GABA receptor was identified in honey bee brain by virtue of its specific binding of [³H]muscimol and its drug specificity. [³H]Muscimol bound with two affinities (K_d1 of 3 nM and K_d2 of 144 nM), comparable to its affinities for binding to mammalian brain. The high-affinity binding was most sensitive to GABA agonists with the following decreasing order of potencies: muscimol>GABA>imidazole acetic acid>DL-GABOB>Zβ-guanidine propionic acid. However, it was insensitive to the antagonist bicuculline, which is potent on [³H]muscimol binding to the mammalian GABA_A receptor. It was also insensitive to baclofen, which is a potent agonist of mammalian GABA_B receptor, as well as to picrotoxinin, pentobarbital, flunitrazepam, and ethyl-β-carboxylate, which bind to allosteric sites in mammalian GABA receptor. The low-affinity [³H]muscimol binding was inhibited with GABA agonists with the following decreasing order of potencies: imidazole acetic acid = β-guanidine propionic acid>dl-GABOB. The two muscimol binding affinities may represent binding to two sites on the same GABA receptor or to two kinds of GABA receptor. The most potent inhibitor of the high-affinity [³H]muscimol binding to honey bee brain was avermectin B_1a (AVM), whose IC₅₀ was 0.01 nM. AVM also inhibited the low-affinity [³H]muscimol binding with an IC₅₀ of 2 μM.     

Uptake patterns of soil-applied 45Ca and 32P in some legume species as influenced by differential trifluralin placement

The effect of localized placement of trifluralin on uptake patterns of soil-applied ⁴⁵Ca in vetch (Vicia sativa L.), pea (Pisum sativum L.) and soybean (Glycine max) and ³²P in vetch and pea was investigated in two soil zones in the roots and in the shoot zone before and after plant emergence. When trifluralin was in the upper root zone severe inhibition of lateral roots occurred as well as a marked decrease in uptake of ⁴⁵Ca and ³²P from this zone. Root growth in the lower zone was unaffected, but uptake of ⁴⁵Ca and ³²P was slightly reduced. Compensatory adventitious root growth as well as a marked increase in uptake of ⁴⁵Ca and ³²P occurred in the shoot zone. Neither root growth nor uptake of ⁴⁵Ca or ³²P in the upper root zone were affected by the presence of trifluralin in the lower root region. When trifluralin was placed in the shoot zone after plant emergence, adven-titious roots on the shoots were inhibited and uptake of ⁴⁵Ca and ³²P was reduced.     

Formamidine structure and detachment of the cattle tick Boophilus microplus

Evaluation of the effectiveness of topically applied chlordimeform, 14 other formamidines, and 5 sulfur-containing related nonformamidine compounds in causing female Boophilus microplus ticks to detach from mice enabled activity to be related to structure. Five compounds were inactive and 15, including 2 sulfur-containing nonformamidines and 1 sulfur-containing formamidine were active at doses ranging from 0.0005 to 2.0 μg/tick. The most active compounds were the N-monomethyl formamidines, BTS-27271 [N′-(2,4-dimethylphenyl)-N-methylformamidine], and C-8520 (demethylchlordimeform). Among the analogs of chlordimeform tested, those with alkyl substitutions at the amino nitrogen decreased in effectiveness in the order, monomethyl (demethylchlordimeform), monoethyl, mono-n-butyl, mono-i-butyl, mono-i-propyl, dimethyl (chlordimeform), and di-n-propyl. Inactive compounds resulted from the replacement of the chlorine at ring position 4 of the aryl moiety of chlordimeform by bromine or hydrogen or by the conversion of the NCH amidino moiety to the NHCSN < thiourea moiety.Detachment due to chlordimeform was antagonized by piperonyl butoxide but that due to its N-monomethyl analog, a known metabolite of chlordimeform in ticks, was synergized by the same compound. These effects on the detaching response parallel those reported elsewhere concerning synergism and antagonism of toxic responses of B. microplus to formamidines.BTS-27271, which was the most effective formamidine in causing detachment after topical application to ticks was moderately effective when injected into mice but its potency relative to chlordimeform was considerably reduced; when sprayed onto cattle BTS-27271 was somewhat more effective in depressing percentage survival of ticks of all stages than chlordimeform.     

Interaction of formamidine pesticides with insect neural octopamine receptors: Effects on radioligand binding and cyclic AMP production

The formamidines chlordimeform, demethylchlordimeform (DCDM), amitraz and BTS-2727l were tested for their respective abilities to inhibit [³H]mianserin binding in membrane preparations of cockroach brain and, in the same tissue, to stimulate octopamine-sensitive receptors coupled to adenylate cyclase. Analysis of [³H]mianserin binding indicated negative cooperativity between two binding sites with respective K_d and B_max values of 3.82 mM and 0.886 pmol (mg protein)^?1 for a high-affinity site and 218 nM and 13.56pmol (mg protein)^?1 for a low-affinity site. DCDM, BTS-27271 and amitraz inhibit [³H]mianserin binding with IC₅₀ values similar to those obtained with octopamine and the μ-adrenergic antagonist, phentolamine, whereas chlordimeform is a poor competitor for the binding sites. Similarly DCDM, BTS-27271 and amitraz elevate cyclic AMP production in brain membrane preparations in a dose-dependent manner with K_a values of 0.32 uM, 1.5 uM and 3.4 uM respectively, whereas chlordimeform was again without effect. The formamidine-mediated responses were fully additive with the evaluation of cyclic AMP obtained using the maximal concentration of dopamine but not with octopamine-mediated increases; thus the formamidine effects appear to be expressed through partial agonism of octopamine receptors that are coupled to adenylate cyclase.     

Effect of chlorsulfuron and 1, 8 naphthalic anhydride on uptake of 45Ca in maize

The effect of chlorsulfuron on uptake of ⁴⁵Ca was studied in maize (Zea mays L. cv. Earliking) plants grown from seeds dusted with 1, 8 naphthalic anhydride (NA). ⁴⁵Ca absorption in sand-grown maize was significantly decreased when chlorsulfuron was applied to the foliage but this was not so when seeds had been dusted with NA. Uptake of ⁴⁵Ca was also reduced when either root or shoot soil zones were separately exposed to chlorsulfuron. When seeds had been dusted with NA, uptake of ⁴⁵Ca from main roots was similar to that of untreated plants, but only when chlorsulfuron was localized in the shoot zone. NA did not counteract the severe reduction in ⁴⁵Ca absorption when chlorsulfuron was localized in the root zone.     

Variability among isolates of <Emphasis Type="Italic">Fusarium udum</Emphasis> and the effect on progression of wilt in pigeonpea

Soft rot of Chinese cabbage is a disease of great economic importance to the State of Pernambuco, Brazil. The present study aimed to evaluate the effect of two calcium sources in different concentrations (calcium nitrate [Ca(NO₃)₂] at 0, 0.15 and 0.3 g l^?1 and calcium chloride (CaCl₂) at 0, 1 and 5 g l^?1) that were applied through two methods (leaf spraying and soil drenching) on the control of soft rot. Further, it aimed to analyze calcium absorption by the plant and to determine calcium’s role in leaf and cell structure using microscopy. Ca(NO₃)₂ applied by both methods was effective in controlling soft rot caused by Pectobacterium carotovorum subsp. carotovorum, as it reduced the disease by up to 48.5 % when sprayed onto the leaves (0.15 g l^?1). A significant increase in the leaf calcium content was observed only in the plants that were sprayed with higher doses of Ca(NO₃)₂ and CaCl₂. In all of the calcium treatments, light microscopy analyses revealed an increased number of chloroplasts and improved structuring of the palisade parenchyma, while transmission electron microscopy analyses revealed an increased cell wall thickness that was especially evident for the 0.15 g l^?1 Ca(NO₃)₂ treatment applied by leaf spraying and soil drenching.     

Chlordimeform: Uncoupling activity against rat liver mitochondria

The pesticide chlordimeform [N′-(4-chloro-o-tolyl)-N,N-dimethylformamidine] at 0.04 μmoles/mg protein uncouples 50% of respiratory-chain phosphorylation of rat liver mitochondria. This uncoupling activity depends on mitochondrial protein concentration and can be reversed either by the addition of bovine serum albumin or by washing. The normal inhibition of state 3 respiration by oligomycin and atractylate is completely reversed by chlordimeform. Uncoupling concentrations of chlordimeform elicit high adenosine triphosphatase activity. This activity is blocked by the above inhibitors of mitochondrial energy-transfer reactions. Evidence is presented which shows that unprotonated chlordimeform is the form effective in uncoupling. It is concluded that chlordimeform is an uncoupling agent with a potency and site of action close to but probably not identical to that of the classical uncoupler 2,4-dinitrophenol.     

Effects of chlordimeform and lindane on monoamine levels in the central nervous system of the american cockroach, Periplaneta americana L

The effects of chlordimeform and lindane on levels of 5-hydroxytryptamine, tryptophan, and N-acetyl dopamine were studied in the cerebral ganglia of the american cockroach, Periplaneta americana. The effects of chlordimeform on nerve cord levels of 5-hydroxytryptamine, tryptophan, dopamine, and octopamine, and the effect of lindane on cerebral ganglia levels of dopamine were also investigated in this species. Topical applications of chlordimeform deplete 5-hydroxytryptamine and tryptophan from the cerebral ganglia whereas levels of n-acetyl dopamine are elevated. The effect of chlordimeform on these compounds is dose-dependent. Similar chlordimeform-induced effects are observed in the nerve cord, and octopamine levels are also depleted in this tissue following treatment with chlordimeform. A biphasic response to chlordimeform is observed in the nerve cord for dopamine levels with a 40% decrease evident after 2 hr and a 30% increase apparent after 6 hr. In contrast to chlordimeform, lindane does not affect 5-hydroxytryptamine and tryptophan levels in the cebral ganglion but low doses of this insecticide effect increases in brain levels of dopamine and n-acetyl dopamine.     

Influence of amidines and anilides on biogenic amine regulatory mechanisms in the bulb mite Rhizoglyphus echinopus

Levels of dopamine (DA) and its precursor 3-(3,4-dihydroxyphenyl)alanine (DOPA) in adult bulb mites Rhizoglyphus echinopus (Fumouze and Robin), after exposure for 1 and 24 h in vials precoated with an acetone solution (1.0 mg ml^?1) of the amidines chlordimeform, clenpyrin, or flubenzimine, were measured by high-performance liquid chromatography with electrochemical detection and compared with those of untreated controls. When mites were exposed to the formamidine chlordimeform, there was an initial increase in levels of DA and DOPA; after 24 h, DOPA levels were still increased, but there was a significant decrease in the levels of DA. Exposure of mites to the cyclic amidines clenpyrin and flubenzimine resulted in decreased levels of DA and DOPA at both time intervals. The effects of amidines and anilides on the metabolism of 2-phenylethylamine (PEA) by whole homogenates of bulb mites were also examined. Degradation of PEA was inhibited by chlordimeform and some of the other formamidines, but not by the cyclic amidines clenpyrin and flubenzimine. Formanilides, that are degradation products of certain formamidines, were the most potent inhibitors of PEA metabolism. These studies provide direct evidence for the interaction of amidines with mite biogenic amines and their regulatory mechanisms.     

The actions of formamidine pesticides on identified central neurones of Helix aspersa

Membrane potential and voltage clamp measurements of the responses to octopamine, dopamine, O-acetylcholine, and the formamidine derivatives chlordimeform (CDM) and N¹-demethylchlordimeform (DCDM) have been made on identified central neurones of Helix aspersa. DCDM and CDM were agonists on both the inhibitory and excitatory octopamine receptors on these neurones but were less potent than octopamine. The direct inhibitory effect of DCDM was antagonised by both phentolamine and cyproheptadine. DCDM was also found to antagonise the effects of octopamine, dopamine and O-acetylcholine on these neurones. The pA₂ values for DCDM against dopamine and O-acetylcholine were 5.2 (±0.1) and 5.35 (±0.15), respectively. The pA₂ value=log[DR-(1/I)], where I is the concentration of the antagonist, and DR=dose ratio =A2/A1; A1 is the dose of agonist required to produce 50% of the maximum response, and A2 is the dose of agonist needed to produce the same degree of response in the presence of the antagonist. It is concluded that DCDM and CDM act specifically on the octopamine receptors of H. aspersa central neurones, both as agonists and antagonists. The antagonist effect of DCDM against dopamine receptors is probably non-specific.