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Human red blood cell acetylcholinesterase inhibition as the appropriate and conservative surrogate endpoint for establishing chlorpyrifos reference dose.

Chlorpyrifos (O,O-diethyl O-(3,5, 6-trichloro-2-pyridinyl)- phosphorothioate) is an organophosphorus (OP) insecticide used for controlling insect pests. Currently, the reference dose (RfD) used by the Environmental Protection Agency (EPA) to establish acceptable human exposure tolerances for chlorpyrifos is based upon inhibition of blood butyrylcholinesterase (BuChE), which is not the target enzyme of chlorpyrifos, and does not play any role in cholinergic transmission. Data are presented showing that inhibition of acetylcholinesterase (AChE) associated with red blood cells (RBC), an enzyme similar to or identical with that in the nervous system, is a more appropriate endpoint on which to base the RfD. Basing an acceptable level of human exposure (e.g., RfD) on inhibition of RBC AChE provides a significant margin of safety, since it is 12- to 14-fold more sensitive as an indicator of chlorpyrifos exposure than the AChE in the most sensitive relevant neurological tissues (brain or retina). Inhibition of RBC AChE activity is consistently exhibited at lower dosages of chlorpyrifos than those required to result in clinical symptoms of OP toxicity, or alterations in cognitive functional responses. There is no unique sensitivity of the fetus or neonates to chlorpyrifos when administered by an appropriate oral dose. Thus, inhibition of RBC AChE activity is an appropriate surrogate measurement of chlorpyrifos exposure and provides a conservative endpoint for establishing appropriate margins of safety for both adults and infants.

Adult↗

Ultrastructure of juvenile hormone-induced degenerating flight muscles in a bark beetle, Ips paraconfusus.

Topical application of 5 microgram of a juvenile hormone analogue (JHA), ZR-615, to female callow adults of Ips paraconfusus induced degeneration of the dorsoventral flight muscles. Within 24 h after JHA-treatment the diameter of the myofibrils was reduced to almost half due to the lysis of the peripheral myofilaments. Mitochondria showed conspicuous degenerative changes like swelling, dissolution of the matrix or presence in the matrix of dense filamentous material or myelin-like figures. Degeneration of the mitochondria seemed to take place inside isolation membranes derived from sarcoplasmic reticulum. A number of granular osmiophilic bodies appeared in the sarcoplasm. Three days after JHA-treatment the muscles were very thin and sheath-like. Most of the mitochondria had already degenerated. The dense sarcoplasm contained numerous crystalline bodies. The granular dense bodies were also more frequent. The myofibrils were comprised of only occasional small bundles of myofilaments. The tubules of the T system enclosed an amorphous material. The nuclei and the tracheal system remained intact but they were crowded due to the decreased volume of the muscle. In some specimens degeneration of the myofibrils and mitochondria was completed by the third day. No sign of degeneration was observed in the flight muscles of acetone treated control insects.

Animals↗

Prothoracicotropic hormone activity in the embryonic brain of the tobacco hornworm, Manduca sexta.

Head segments and brains were extirpated from embryos of the tobacco hornworm, Manduca sexta, extracted and the resulting extracts assayed for prothoracicotropic hormone (PTTH) activity on prothoracic glands from day 3 fifth instar larvae and day 0 pupae. Dose-response curves were generated and indicated the presence of PTTH activity in embryonic brains and head segments, suggesting a role(s) for this neurohormone during embryogenesis. Maximal PTTH activity was found in brains from embryos 117 h post-oviposition, just prior to hatching, but activity was also noted in head segments as early as 24 h post-oviposition. These data on PTTH and those on ecdysteroids and juvenile hormones in embryos suggest that these 3 classes of hormones which control insect post-embryonic development, may also be involved in the regulation of developmental processes in the embryo.

Animals↗

Physical characteristics of the cerebral big prothoracicotropic hormone from Manduca sexta.

The prothoracicotropic hormones (PTTHs) are cerebral peptides that control insect postembryonic development by stimulating the prothoracic glands to synthesize ecdysteroids. In Manduca sexta, the tobacco hornworm, two classes of PTTH are distinguished by their M(r), small (ca. 7 kDa) and big PTTH (ca. 25-30 kDa). Little is known about the physical nature of the PTTHs and this study takes a first step towards defining characteristics of the Manduca big PTTH. The neurohormone has a Stokes radius of 2.59 nm and a sedimentation coefficient of 2.76 S. Based on these data, an M(r) of 29,443.7 and an f/fo of 1.27 were calculated. Combined, the physical data reveal Manduca big PTTH is an asymmetrical acidic homodimeric peptide with intra- and intermolecular disulfide bonds.

Animals↗

Biosynthesis and distribution of insect-molting hormones in plants--a review.

Insect-molting hormones, phytoecdysteroids, have been reported to occur in over 100 plant families. Plants, unlike insects, are capable of the biosynthesis of ecdysteroids from mevalonic acid, and in several cases the biosynthesis of phytoecdysteroids was also demonstrated to proceed via sterols. Spinacia oleracea (spinach) biosynthesizes polypodine B and 20-hydroxyecdysone, which is the predominant insect-molting hormone found in plant species. The onset of ecdysteroid production in spinach requires the appropriate ontogenetic development within the plant, which is related to leaf development. In spinach, lathosterol is the biosynthetic precursor to ecdysone and 20-hydroxyecdysone. Phosphorylated ecdysteroid intermediates, particularly ecdysone-3-phosphate, are required during biosynthesis. Polyphosphorylated forms of ecdysteroids are putative regulatory components of the pathway. During spinach development, the 20-hydroxyecdysone is transported from the sites of biosynthesis to the apical regions. An analysis of the physiological data available suggests that different species may synthesize ecdysteroids in various organs and distribute these ecdysteroids to other sites. Annual plants appear to concentrate ecdysteroids in the apical regions, including flowers and seeds. Perennial plants may recycle their ecdysteroids between their deciduous and their perennial organs over the growing season. Further investigations of ecdysteroid biosynthesis and physiology within plants will be required before an acceptable system can be designed to test phytoecdysteroid effectiveness in vivo against insect herbivory.

Ecdysteroids↗