Pesticidal compounds from higher plants

Higher plants offer an excellent source of biologically active natural products. Over the centuries numerous plants have been exploited as sources of insecticides, but nowadays traditional botanical insecticides play only a minor role in world agriculture. Nevertheless, plant natural products still have enormous potential to inspire and influence modern agrochemical research. Few plant natural products will ever reach the market as products per se, but others will provide lead structures for programmes of synthetic chemistry and hopefully follow the success story of the synthetic pyrethroids. Structurally complex compounds, which are not amenable to synthetic chemistry programmes, may also have a role to play by validating new modes of action for pesticides. Examples are presented of compounds exhibiting insecticidal, fungicidal and herbicidal effects. Consideration is also given to the development of screening programmes to detect new compounds with interesting biological properties. Careful experimental design and thorough recording of procedures and data are crucial to success. Badly designed programmes afford only weakly active compounds or show effects which cannot be reproduced at a later date. Natural product chemistry, whether based on higher plants, micro-organisms or other sources, is a very difficult science, but there is little doubt that dedicated research will eventually be rewarded with exciting new lead structures for industrial application.     

The effects of a protein deficiency on egg composition

Eggs were collected from groups of pullets receiving 9, 14 or 20 per cent dietary protein and the component parts were separated and weighed. The reduction in egg weight associated with protein deficiency was found to be due to reductions in all components, but yolk and shell weight changed proportionately less than total weight and albumen weight proportionately more. The resulting increase in percentage yolk was found to be of similar magnitude to that expected from a consideration of the relationship between egg size and yolk size in birds receiving adequate protein. In a similar way the increased proportion of shell apparently reflected only the relationship between egg size and surface area in eggs collected when the birds were 52 weeks of age but at 67 weeks a real increase in shell thickness on the deficient diet was also indicated.     

Studies on a fluorescent insect growth regulator metabolite complex with house-fly microsomal cytochrome P-450

The fluorescent insect growth regulator 5[[[5-(dimethylamino)-1-naphthalenyl]amino]-1,3-benzodioxole (DNSAB) forms a metabolite complex with house-fly microsomal cytochrome P-450. Formation of the metabolite complex is dependent on the presence of NADPH and O₂; NADH supports the reaction at a reduced rate. The presence of antibodies to house-fly cytochrome c (P-450) reductase in reaction mixtures inhibits the complex formation, indicating that the reductase is necessary for transfer of electrons from NADPH to cytochrome P-450 to complete the reaction. In the oxidized form, the metabolite complex has a single absorbance maximum at 431 nm, whereas the reduced form has two absorbance maxima at 426 (major) and 455 nm (minor). The pH of the media affects the extinction of the 426- and 455-nm Soret bands; increased pH decreases the extinction of the 426-nm band and increases the extinction of 455-nm band. Formation of the DNSAB metabolite-cytochrome P-450 complex decreases the amount of CO-reactive cytochrome P-450 by 24%. The metabolite complex is not dissociable by treatment with ferricyanide or by using centrifugation techniques. Dissociation is accomplished by addition of DNSAB to the oxidized metabolite complex. Kinetic analysis of the complex formation gives apparent K_m and V_max values at 2.55 ± 1.0 μM and 1.1 ± 0.4 × 10^?2 ΔA min^?1 nmol^?1 cytochrome P-450, respectively. Addition of juvenile hormone [(E,E)-cis-methyl-10,11-epoxy-7-ethyl-3,11-dimethyl-2,6-tridecadienoate; JH] to the reaction medium competitively inhibits the formation of the metabolite complex giving an inhibition constant of 16 μM. DNSAB synergized the lethal effects of JH against Aedes aegypti larvae threefold; however, JH did not synergize DNSAB. These data suggest that DNSAB may acquire its hormonal qualities by complexing a species of cytochrome P-450 that metabolizes JH, thereby prolonging the in vivo lifetime of this hormone.     

Detection, quantification, and pharmacokinetics of furosemide and its effects on urinary specific gravity following IV administration to horses

Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.     

Effect of furosemide on urine specific gravity and osmolality in thoroughbred racehorses

Postrace urine samples from thoroughbred horses were examined to compare osmolality and specific gravity between horses treated with furosemide and those not treated. Samples were assigned to groups in relation to reported medication (furosemide) status, race finish position, and distance of race. Urine osmolality was significantly (P <.05) lower in samples from horses treated with furosemide when compared with untreated horses. Specific gravity determinations are less precise at measuring urine osmolality at lower levels (1.01 g/ml or less). The measurement of osmolality is a superior method for determining the urine solute concentration and facilitating the regulation of furosemide.     

Seasonal interaction of serum vitamin D concentration and bone density in alpacas

OBJECTIVE: To evaluate temporal changes in bone mineral density associated with seasonal variation in serum vitamin D, calcium, and phosphorus concentrations in alpacas. ANIMALS: 5 healthy mature neutered male alpacas. PROCEDURE: Metacarpal bone mineral density was measured at 4 times during a year. Each time alpacas were weighed, blood was collected for determination of serum calcium, phosphorus, and vitamin D concentrations, and samples of feed were analyzed for nutrient content. Vitamin D status was determined by use of an assay that measured serum 25-hydroxycalciferol concentration. Effects of changes in serum vitamin D, calcium, and phosphorus concentration and body weight with season on bone mineral density were determined. RESULTS: Bone mineral density, body weight, and serum vitamin D and phosphorus concentrations varied with season. Bone mineral density, serum vitamin D concentration, and body weight also varied among individual alpacas. Serum vitamin D concentration was lower in January than the previous October and increased from May to the following September. The decrease in bone mineral density lagged behind the decrease in serum vitamin D concentration and was lower in May, compared with the previous October. Body weight was lower in May than the previous October or following September. Solar radiation was highest in July and lowest in December. CONCLUSIONS AND CLINICAL RELEVANCE: Seasonal changes in bone mineral density are associated with changes in serum vitamin D concentrations in alpacas. Changes in bone mineral density associated with a decline in serum vitamin D concentration may predispose some alpacas to developing fractures minimal trauma.