Glutathione transferase activities toward herbicides used selectively in soybean

Using extracts from suspension-cultured cells of soybean (Glycine max cv. Mandarin) as a source of active enzymes, the activities of glutathione transferases (GSTs) catalysing the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and selective herbicides were determined to be in the order CDNB≫ fomesafen>metolachlor=acifluorfen>chlorimuron-ethyl. GST activities showed a thiol dependence in a substrate-specific manner. Thus, GST activities toward acifluorfen and fomesafen were greater when homoglutathione (hGSH), the endogenously occurring thiol in soybean, was used as the co-substrate rather than glutathione (GSH). Compared with GSH, hGSH addition either reduced or had no effect on GST activities toward other substrates. In the absence of enzyme, the rates of hGSH conjugation with acifluorfen, chlorimuron-ethyl and fomesafen were negligible, suggesting that rapid hGSH conjugation in soybean must be catalysed by GSTs. GST activities were subsequently determined in 14-day-old plants of soybean and a number of annual grass and broadleaf weeds. GST activities of the plants were then related to observed sensitivities to post-emergence applications of the four herbicides. When enzyme activity was expressed on a mg^-1 protein basis, all grass weeds and Abutilon theophrasticontained considerably higher GST activity toward CDNB than soybean. With fomesafen as the substrate, GST activities were determined to be in the order soybean≫Echinochloa crus-galli>Digitaria sanguinalis>Sorghum halepense=Setaria faberi with none of the broadleaf weeds showing any activity. This order related well to the observed selectivity of fomesafen, with the exception of A. theophrasti, which was partially tolerant to the herbicide. Using metolachlor as the substrate the order of the GST activities was soybean>A. theophrasti≫S. halepense>Amaranthus retroflexus>Ipomoea hederacea, with the remaining species showing no activity. GST activities toward metolachlor correlated well with the selectivity of the herbicide toward the broadleaf weeds but not toward the grass weeds. Acifluorfen and chlorimuron-ethyl were selectively active on these species, but GST activities toward these herbicides could not be detected in crude extracts from whole plants. © 1997 SCI     

Environmental fate of mineral,vegetable and transesterified vegetable oils

Vegetable oils, synthetic esters (including transesterified oils) and mineral oils are the main classes of oil used in pesticide formulations. Biodegradation is a major route for the removal of oils from soil systems. Most of the oils used in pesticide formulations are degraded substantially in the laboratory tests that are used to assess aquatic biodegradability. The susceptibility of different oils to biodegradation can be explained in terms of the metabolic capacity of common microorganisms. Fewer soil biodegradability tests have been carried out with oils, but the available data suggest that the mode of degradation is not very different from that in aquatic systems. Supplements of inorganic nutrients (in particular nitrogen) are needed to stimulate microbial activity in soils containing the high concentrations of oil that can be encountered in the event of a spill. However, oils are applied at such low rates in pesticide form illations (maximum of 5g oil m^? soil) that nutrient availability is unlikely to affect the rate of biodegradation in the field.     

Microanatomic features of pancreatic islets and immunolocalization of glucose transporters in tissues of llamas and alpacas

OBJECTIVE: To describe the microanatomic features of pancreatic islets and the immunohistochemical distribution of glucose transporter (GLUT) molecules in the pancreas and other tissues of New World camelids. ANIMALS: 7 healthy adult New World camelids, 2 neonatal camelids with developmental skeletal abnormalities, and 2 BALB/c mice. PROCEDURE: Samples of pancreas, liver, skeletal muscle, mammary gland, brain, and adipose tissue were collected postmortem from camelids and mice. Pancreatic tissue sections from camelids were assessed microscopically. Sections of all tissues from camelids and mice (positive control specimens) were examined after staining with antibodies against GLUT-1, -2, -3, and -4 molecules. RESULTS: In camelids, pancreatic islets were prominent and lacked connective tissue capsules. Numerous individual endocrine-type cells were visible distant from the islets. Findings in neonatal and adult tissues were similar; however, the former appeared to have more non-islet-associated endocrine cells. Via immunostaining, GLUT-2 molecules were detected on pancreatic endocrine cells and hepatocytes in camelids, GLUT-1 molecules were detected on the capillary endothelium of the CNS, GLUT-3 molecules were detected throughout the gray matter, and GLUT-4 molecules were not detected in any camelid tissues. Staining characteristics of neonatal and adult tissues were similar. CONCLUSIONS AND CLINICAL RELEVANCE: In New World camelids, microanatomic features of pancreatic islets are similar to those of other mammals. Data suggest that the poor glucose clearance and poor insulin response to hyperglycemia in adult camelids cannot be attributed to a lack of islet cells or lack of GLUT molecules on the outer membrane of those cells.     

Serum 17-alpha-hydroxyprogesterone and corticosterone concentrations in dogs with nonadrenal neoplasia and dogs with suspected hyperadrenocorticism

OBJECTIVE: To assess serum 17-alpha-hydroxyprogesterone (17OHP) and corticosterone concentrations in dogs with nonadrenal neoplasia and dogs being screened for hyperadrenocorticism. DESIGN: Prospective study. ANIMALS: 16 clinically normal dogs, 35 dogs with nonadrenal neoplasia, and 127 dogs with suspected hyperadrenocorticism. PROCEDURE: ACTH stimulation tests were performed in all dogs. Baseline serum cortisol and corticosterone concentrations were measured in the healthy dogs; baseline serum cortisol concentration and ACTH-stimulated cortisol, corticosterone, and 17OHP concentrations were measured in all dogs. Endogenous plasma ACTH concentration was also measured before administration of ACTH in dogs with neoplasia. RESULTS: In 35 dogs with neoplasia, 31.4% had high serum 17OHP concentration and 22.9% had high serum corticosterone concentration. Of the 127 dogs with suspected hyperadrenocorticism, 59 (46.5%) had high ACTH-stimulated cortisol concentrations; of those, 42 of 59 (71.2%) and 32 of 53 (60.4%) had high serum 17OHP and corticosterone concentrations, respectively. Of dogs with serum cortisol concentration within reference range after ACTH administration, 9 of 68 (13.2%) and 7 of 67 (10.4%) had high serum 17OHP and corticosterone concentrations, respectively. In the dogs with neoplasia and dogs suspected of having hyperadrenocorticism, post-ACTH serum hormone concentrations were significantly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Serum concentrations of 17OHP or corticosterone after administration of ACTH may be high in dogs with nonadrenal neoplasia and no evidence of hyperadrenocorticism. Changes in serum 17OHP or corticosterone concentrations after administration of ACTH are proportionate with changes in cortisol concentration.     

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.     

Successful drug development despite adverse preclinical findings part 1: processes to address issues and most important findings

Unexpected adverse preclinical findings (APFs) are not infrequently encountered during drug development. Such APFs can be functional disturbances such as QT prolongation, morphological toxicity or carcinogenicity. The latter is of particular concern in conjunction with equivocal genotoxicity results. The toxicologic pathologist plays an important role in recognizing these effects, in helping to characterize them, to evaluate their risk for man, and in proposing measures to mitigate the risk particularly in early clinical trials. A careful scientific evaluation is crucial while termination of the development of a potentially useful drug must be avoided. This first part of the review discusses processes to address unexpected APFs and provides an overview over typical APFs in particular classes of drugs. If the mode of action (MoA) by which a drug candidate produces an APF is known, this supports evaluation of its relevance for humans. Tailor-made mechanistic studies, when needed, must be planned carefully to test one or several hypotheses regarding the potential MoA and to provide further data for risk evaluation. Safety considerations are based on exposure at no-observed-adverse-effect levels (NOAEL) of the most sensitive and relevant animal species and guide dose escalation in clinical trials. The availability of early markers of toxicity for monitoring of humans adds further safety to clinical studies. Risk evaluation is concluded by a weight of evidence analysis (WoE) with an array of parameters including drug use, medical need and alternatives on the market. In the second part of this review relevant examples of APFs will be discussed in more detail.     

Aerobic microbial respiration in 86-million-year-old deep-sea red clay

Microbial communities can subsist at depth in marine sediments without fresh supply of organic matter for millions of years. At threshold sedimentation rates of 1 millimeter per 1000 years, the low rates of microbial community metabolism in the North Pacific Gyre allow sediments to remain oxygenated tens of meters below the sea floor. We found that the oxygen respiration rates dropped from 10 micromoles of O(2) liter(-1) year(-1) near the sediment-water interface to 0.001 micromoles of O(2) liter(-1) year(-1) at 30-meter depth within 86 million-year-old sediment. The cell-specific respiration rate decreased with depth but stabilized at around 10(-3) femtomoles of O(2) cell(-1) day(-1) 10 meters below the seafloor. This result indicated that the community size is controlled by the rate of carbon oxidation and thereby by the low available energy flux.     

Lepidium latifolium: plant nutrient competition-soil interactions

Exotic weeds are invading rangelands of the western United States at unprecedented rates. Understanding plant-soil relationships and competitive interactions of invasive weeds is crucial in long-term control strategies. In a greenhouse experiment, we investigated the influence of soil nutrient depletion on plant growth and plant competition between the exotic invasive weeds, Lepidium latifolium (invading wetlands) and Bromus tectorum (invading a multitude of habitats). Plants were grown individually and in combination until L. latifolium flowered, then roots and aboveground mass were harvested. Soil in individual pots was homogenized, subsamples collected for nutrient analyses, and the soil was re-planted to the same species, grown, and harvested twice more for a total of three growth cycles. As nutrient supplying capacity of the soil declined through growth cycles, aboveground mass of L. latifolium decreased significantly (PА.05) and growth potential of B. tectorum surpassed that of L. latifolium. Only bicarbonate-extractable soil ortho-P positively correlated with plant mass of L. latifolium. A separate experiment demonstrated that L. latifolium has a narrow window of soil water potentials for optimal growth; greatest growth at -20 kPa with significantly declining growth at saturation and -400 kPa. Our data suggest that L. latifolium primarily invades wetlands because the high soil water content reduces tortuosity and allows efficient transport of nutrients to this sparsely rooted species. When soil moisture and/or the nutrient supplying capacity of the soil declines, plants with greater root density can out-compete L. latifolium. Monocultural stands of L. latifolium may be self-limiting in time as available nutrients, particularly P, are biocycled to drier upper soil layers.     

Effects of testicular biopsy in clinically normal bulls

OBJECTIVE: To determine whether testicular needle biopsy is detrimental to testicular function in clinically normal bulls. DESIGN: Prospective study. ANIMALS: 6 mixed-breed mature bulls. PROCEDURE: A randomly selected testicle from each bull was biopsied with a 14-gauge needle biopsy instrument. Bulls were then evaluated over a 90-day period for changes in scrotal temperature and thermal patterns, ultrasonographic appearance, and quality of spermatozoa. At the end of the 90-day study, bulls were castrated, and testicles were examined grossly and histologically. RESULTS: Changes were detected in scrotal temperatures and thermal patterns and in the breeding soundness examination results during the first 2 weeks of the study. However, there were no long-term changes in semen quality over the course of the experiment. Hyperechoic areas were detected on ultrasonographic examination and corresponded to the areas of penetration by the biopsy instrument. Microscopic lesions that were indicative of testicular dysfunction were not found. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that testicular biopsy is a safe procedure in bulls. Testicular biopsy could possibly be used to further examine bulls that have less than satisfactory results for breeding soundness examinations.     

Biochemical response of grapevine variety ‘Chardonnay’ (Vitis vinifera L.) to infection with grapevine yellows (Bois noir)

This study was carried out on the leaves of phytoplasma susceptible grapevine variety ??Chardonnay?? (Vitis vinifera L.), and included research of the alterations in chlorophyll and carotenoid contents, contents of phenol compounds and in related enzymes activity in the phenylpropanoid pathway during the Bois noir (BN) infection. Phytoplasma-infected leaves showed reduced contents of chlorophylls and carotenoids, which promoted their susceptibility to oxidative reactions. Furthermore, modulation of flavonoid biosynthesis occurred in infected leaves, leading to an increased activity of phenylalanine ammonia lyase, chalcone synthase/chalcone isomerase, flavanone 3-hydroxylase and polyphenoloxidase, but to a decreased peroxidase activity. Phytoplasma infection led to an increase of the contents of hydroxycinnamic acids (caftaric acid, sinapic acid glucose derivate and coutaric acid), flavanols (procyanidin B1, procyanidin dimer 3, catechin, epicatechin) and flavonols (quercetin 3-O-glucuronide, quercetin 3-O-glucoside) especially in the period up to vérasion. The study demonstrated that at certain phenological key-stages infection with phytoplasma (BN) induced different alterations in enzyme activities and in the contents of biochemical compounds from primary and secondary metabolism.