N., and Bakry, N. M. (2006)

White, Brandy J. and Harmon, H. James (2005). Optical solid-state detection of organophosphates using organophosphorus hydrolase: Selected Papers from the Eighth World Congress on Biosensors, Part II. Biosensors and Bioelectronics 20: 1977-1983.
Chem Codes: Chemical of Concern: DZ Rejection Code: METHODS.

We have developed a sensor surface for optical detection of organophosphates based on reversible inhibition of organophosphorus hydrolase (OPH) by copper complexed meso-tri(4-sulfonato phenyl) mono(4-carboxy phenyl) porphyrin (CuC1TPP). OPH immobilized onto glass microscope slides retains catalytic activity for more than 232 days. CuC1TPP is a reversible, competitive inhibitor of OPH, binding at the active site of the immobilized enzyme. The absorbance spectrum of the porphyrin-enzyme complex is measured via planar waveguide evanescent wave absorbance spectroscopy using a blue LED as a light source and an Ocean Optics USB2000 as the spectrophotometer. The characteristics of the absorbance spectrum of CuC1TPP are specific and different when the porphyrin is bound to the enzyme or is bound non-specifically to the surface of the slide. Addition of a substrate of OPH such as one of the organophosphates paraoxon, coumaphos, diazinon, or malathion displaces the porphyrin from the enzyme resulting in reduced absorbance intensity at 412 nm. Absorbance changes at 412 nm show log-linear dependence on substrate concentration. Paraoxon concentrations between 7 parts per trillion (ppt) and 14 parts per million (ppm) were investigated and a 3:1 S/N detection limit of 7 ppt was determined. Concentrations of 700 ppt to 40 ppm were investigated for diazinon, malathion, and coumaphos with detection limits of 800 ppt, 1 part per billion, and 250 ppt, respectively. This optical technique does not require the addition of reagents or solutions other than the sample and absorbance spectra can be collected in less than 6 s. Porphyrin/ Organophosphate hydrolase/ Competitive inhibitor/ Absorbance spectroscopy/ Planar waveguide

White, Brandy J., Legako, J. Andrew, and Harmon, H. James (2003). Extended lifetime of reagentless detector for multiple inhibitors of acetylcholinesterase: Selected papers from the Seventh World Congress on Biosensors Kyoto, Japan 15-17 May 2002. Biosensors and Bioelectronics 18: 729-734.
Chem Codes: Chemical of Concern: DZ Rejection Code: METHODS.

Competitive inhibitors of acetylcholinesterase (AChE) are detected using an evanescent wave technique to monitor changes in the absorbance spectrum of an AChE-monosulfonate tetraphenyl porphyrin (TPPS1) complex immobilized on the surface of a glass slide. In this technique, porphyrin is displaced from the AChE active site by the inhibitor. The loss in absorbance intensity of the characteristic absorbance peak for the AChE-TPPS1 complex at 446 nm is linearly dependent on the log of the inhibitor concentration. This technique yields detection limits at 3:1 S/N of 37 ppt for eserine, 50 ppt for galanthamine, 100 ppt for scopolamine, 250 ppt for tetracaine, 45 ppt for diazinon, and 83 ppb for Triton X-100. When stored under vacuum, the enzymatic lifetime of the immobilized AChE surface is greater than 73 days while the responsive lifetime of the immobilized AChE-TPPS1 surface is currently 49 days. Acetylcholinesterase/ Porphyrin/ Optical sensor/ Biosensor/ Evanescent wave/ Acetylcholinesterase inhibitor

WHITE PF (1995). BIOLOGICAL CONTROL OF MUSHROOM PESTS AN EVALUATION. ELLIOTT, T. J. (ED.). MUSHROOM SCIENCE, VOL. 14. 1-2. SCIENCE AND CULTIVATION OF EDIBLE FUNGI, VOLS. 1 AND 2; 14TH INTERNATIONAL CONGRESS, OXFORD, ENGLAND, UK, SEPTEMBER 17-22, 1995. XXIV+472P.(VOL. 1); XI+501P.(VOL. 2) A. A. BALKEMA: ROTTERDAM, NETHERLANDS; BROOKFIELD, VERMONT, USA. ISBN 90-5410-570-4(SET); ISBN 90-5410-571-2(VOL. 1); ISBN 90-5410-572-0(VOL. 2).; 14 (1-2). 1995. 475-484.
Chem Codes: Chemical of Concern: DZ Rejection Code: ABSTRACT.

BIOSIS COPYRIGHT: BIOL ABS. RRM BOOK CHAPTER MEETING POSTER PREDATORS PARASITES PATHOGENS PHEROMONES KAIROMONES REPELLENTS ANTI-FEEDANTS GENETIC CONTROL INSECT CONTROL PRODUCT AVAILABILITY INDUSTRY PROSPECTS Congresses/ Biology/ Genetics/ Cytogenetics/ Animals/Genetics/ Behavior, Animal/ Animal Communication/ Behavior/ Ecology/ Animals/ Ecology/ Biochemistry/ Nutrition/ Nutritional Status/ Endocrine Glands/ Microbiological Techniques/ Vegetables/ Herbicides/ Pest Control/ Pesticides/ Arachnida/ Entomology/Economics/ Plants/ Arachnida/ Entomology/Economics/ Pest Control, Biological/ Animal/ Animals, Laboratory/ Animals, Wild/ Parasitic Diseases/Veterinary/ Animal/ Disease/ Insects/Parasitology/ Basidiomycota/ Insects

Whitehead, A. G. and Turner, S. J. (1998). Management and Regulatory Control Strategies for Potato Cyst Nematodes (Globodera rostochiensis and Globodera pallida). In: R.J.Marks and B.B.Brodie (Eds.), Potato Cyst Nematodes: Biology, Distribution and Control, Cab Int., Wallingford, England 135-152.
Chem Codes: EcoReference No.: 76547
Chemical of Concern: NaN3,MB,ASCN,DZ Rejection Code: REVIEW.

Whitmore, D. H. Jr. and Hodges, D. H. Jr. (1978). In Vitro Pesticide Inhibition of Muscle Esterases of the Mosquitofish, Gambusia affinis. Comp.Biochem.Physiol.C 59: 145-149.

Chem Codes: EcoReference No.: 85460
Chemical of Concern: DZ Rejection Code: IN VITRO.

Whitney, W. K. (1967). Laboratory Tests with Dursban and Other Insecticides in Soil. J.Econ.Entomol. 60: 68-74.

EcoReference No.: 69734

Chemical of Concern: DZ; Habitat: T; Rejection Code: TARGET(DZ).

WHO (1985). SPECIFICATIONS FOR PESTICIDES USED IN PUBLIC HEALTH INSECTICIDES MOLLUSCICIDES REPELLENTS METHODS SIXTH EDITION. WHO. SPECIFICATIONS FOR PESTICIDES USED IN PUBLIC HEALTH: INSECTICIDES, MOLLUSCICIDES, REPELLENTS, METHODS, SIXTH EDITION. 384P. WHO: GENEVA, SWITZERLAND. ILLUS. PAPER. ISBN 92-4-156084-3.; 0 (0). 1985 (RECD. 1986). 384P.

Chem Codes: Chemical of Concern: DZ Rejection Code: NO TOX DATA.

BIOSIS COPYRIGHT: BIOL ABS. RRM BOOK FAO VECTOR CONTROL DDT BHC TRIFENMORPH METHOXYCHLOR PYRETHRUM DIAZINON MALATHION TRICHLORFON FENTHION DICHLORVOS FENITROTHION PROPOXUR TEMEPHOS BROMOPHOS CHLORPYRIFOS JODFENPHOS BENDIOCARB DELTAMETHRIN NICLOSAMIDE TECHNICAL N N DIETHYL-M-TOLUAMIDE Legislation/ Organization and Administration/ Biology/ Animals, Wild/ Conservation of Natural Resources/ Ecology/ Biochemistry/ Poisoning/ Animals, Laboratory/ Disease Vectors/ Disinfection/ Pest Control/ Pesticides/ Disease Vectors/ Herbicides/ Pest Control/ Pesticides/ Animals/ Arachnida/ Entomology/Economics/ Pest Control/ Arachnida/ Entomology/Economics/ Insecticides/ Pest Control/ Pesticides/ Animal/ Animals, Laboratory/ Animals, Wild/ Parasitic Diseases/Veterinary/ Arachnida/ Disease Vectors/ Entomology/ Sanitation

Whyatt, R. M., Camann, D. E., Kinney, P. L., Reyes, A., Ramirez, J., Dietrich, J., Diaz, D., Holmes, D., and Perera, F. P. ( Residential Pesticide Use during Pregnancy among a Cohort of Urban Minority Women. Environmental Health Perspectives [Environ. Health Perspect.]. Vol. 110, no. 5, pp. 507-514. May 2002.
Chem Codes: Chemical of Concern: PPB Rejection Code: HUMAN HEALTH.

Residential pesticide use is widespread in the United States. However, data are limited specific to use among minority populations. Nor are data available on the extent of pesticide exposure resulting from residential use during pregnancy. We have gathered questionnaire data on pesticide use in the home during pregnancy from 316 African-American and Dominican women residing in northern Manhattan and the South Bronx. Additionally, 72 women underwent personal air monitoring for 48 hr during their third trimester of pregnancy to determine exposure levels to 21 pesticides (19 insecticides and 2 fungicides). Of the women questioned, 266 of 314 (85%) reported that pest control measures were used in the home during pregnancy; 111 of 314 (35%) reported that their homes were sprayed by an exterminator, and of those, 45% said the spraying was done more than once per month. Most ( greater than or equal to 90%) of the pesticide was used for cockroach control. Use of pest control measures increased significantly with the level of housing disrepair reported. Of the women monitored, all (100%) had detectable levels of three insecticides: the organophosphates diazinon (range, 2.0-6,010 ng/m super(3)) and chlorpyrifos (range, 0.7-193 ng/m super(3)) and the carbamate propoxur (range, 3.8-1,380 ng/m super(3)), as well as the fungicide o-phenylphenol (range, 5.7-743 ng/m super(3)). We also frequently detected the following four insecticides (47-83% of samples) but at lower concentrations: the pyrethroid trans-permethrin, piperonyl butoxide (an indicator of exposure to pyrethrins), and the organochlorines 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane and chlordane. Thirty percent of the women had detectable levels of all eight pesticides. Exposures were generally higher among African Americans than among Dominicans. We detected other pesticides in less than or equal to 10% of samples. Results show widespread prenatal pesticide use among minority women in this cohort. Diazinon exposures for some women may have exceeded health-based levels, and our findings support recent federal action to phase out residential use of this insecticide. Fungicides/ Insecticides/ Pregnancy/ Pesticides/ Residential areas/ Urban areas/ Ethnic groups/ Socioeconomics/ Socio-economic aspects Classification: P 6000 TOXICOLOGY AND HEALTH; H 5000 Pesticides; X 24133 Metabolism

Whyatt, R. M., Camann, D. E., Kinney, P. L., Reyes, A., Ramirez, J., Dietrich, J., Diaz, D., Holmes, D., and Perera, F. P. (2002). Residential Pesticide Use during Pregnancy among a Cohort of Urban Minority Women. Environmental Health Perspectives [Environ. Health Perspect.]. Vol. 110, no. 5, pp. 507-514. May 2002.
Chem Codes: Chemical of Concern: DZ Rejection Code: HUMAN HEALTH.

ISSN: 0091-6765

Descriptors: Fungicides
Descriptors: Insecticides
Descriptors: Pregnancy
Descriptors: Pesticides
Descriptors: Residential areas
Descriptors: Urban areas
Descriptors: Ethnic groups
Descriptors: Socioeconomics
Descriptors: Socio-economic aspects
Abstract: Residential pesticide use is widespread in the United States. However, data are limited specific to use among minority populations. Nor are data available on the extent of pesticide exposure resulting from residential use during pregnancy. We have gathered questionnaire data on pesticide use in the home during pregnancy from 316 African-American and Dominican women residing in northern Manhattan and the South Bronx. Additionally, 72 women underwent personal air monitoring for 48 hr during their third trimester of pregnancy to determine exposure levels to 21 pesticides (19 insecticides and 2 fungicides). Of the women questioned, 266 of 314 (85%) reported that pest control measures were used in the home during pregnancy; 111 of 314 (35%) reported that their homes were sprayed by an exterminator, and of those, 45% said the spraying was done more than once per month. Most ( greater than or equal to 90%) of the pesticide was used for cockroach control. Use of pest control measures increased significantly with the level of housing disrepair reported. Of the women monitored, all (100%) had detectable levels of three insecticides: the organophosphates diazinon (range, 2.0-6,010 ng/m super(3)) and chlorpyrifos (range, 0.7-193 ng/m super(3)) and the carbamate propoxur (range, 3.8-1,380 ng/m super(3)), as well as the fungicide o-phenylphenol (range, 5.7-743 ng/m super(3)). We also frequently detected the following four insecticides (47-83% of samples) but at lower concentrations: the pyrethroid trans-permethrin, piperonyl butoxide (an indicator of exposure to pyrethrins), and the organochlorines 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane and chlordane. Thirty percent of the women had detectable levels of all eight pesticides. Exposures were generally higher among African Americans than among Dominicans. We detected other pesticides in less than or equal to 10% of samples. Results show widespread prenatal pesticide use among minority women in this cohort. Diazinon exposures for some women may have exceeded health-based levels, and our findings support recent federal action to phase out residential use of this insecticide.
Language: English
English
Publication Type: Journal Article
Classification: P 6000 TOXICOLOGY AND HEALTH
Classification: H 5000 Pesticides
Classification: X 24133 Metabolism
Subfile: Toxicology Abstracts; Health & Safety Science Abstracts; Pollution Abstracts

Whyatt, R. M., Rauh, V., Barr, D. B., Camann, D. E., Andrews, H. F., Garfinkel, R., Hoepner, L. A., Diaz, D., Dietrich, J., Reyes, A., Tang, D., Kinney, P. L., and Perera, F. P. (2004). Prenatal Insecticide Exposures and Birth Weight and Length among an Urban Minority Cohort. Environmental Health Perspectives [Environ. Health Perspect.]. Vol. 112, no. 10, pp. 1125-1132. Jul 2004.

Chem Codes: Chemical of Concern: DZ Rejection Code: HUMAN HEALTH.

ISSN: 0091-6765

Descriptors: Pesticides
Descriptors: Insecticides
Descriptors: Birth weight
Descriptors: Pregnancy
Descriptors: Prenatal experience
Descriptors: Diazinon
Descriptors: Chlorpyrifos
Descriptors: Urban populations
Descriptors: Fetuses
Descriptors: Organophosphates
Descriptors: Urban areas
Descriptors: USA, New York
Abstract: We reported previously that insecticide exposures were widespread among minority women in New York City during pregnancy and that levels of the organophosphate chlorpyrifos in umbilical cord plasma were inversely associated with birth weight and length. Here we expand analyses to include additional insecticides (the organophosphate diazinon and the carbamate propoxur), a larger sample size (n = 314 mother-newborn pairs), and insecticide measurements in maternal personal air during pregnancy as well as in umbilical cord plasma at delivery. Controlling for potential confounders, we found no association between maternal personal air insecticide levels and birth weight, length, or head circumference. For each log unit increase in cord plasma chlorpyrifos levels, birth weight decreased by 42.6 g [95% confidence interval (CI), -81.8 to -3.8, p = 0.03] and birth length decreased by 0.24 cm (95% CI, -0.47 to -0.01, p = 0.04). Combined measures of (In)cord plasma chlorpyrifos and diazinon (adjusted for relative potency) were also inversely associated with birth weight and length (p < 0.05). Birth weight averaged 186.3 g less (95% CI, -375.2 to -45.5) among newborns with the highest compared with lowest 26% of exposure levels (p = 0.01). Further, the associations between birth weight and length and cord plasma chlorpyrifos and diazinon were highly significant (p less than or equal to 0.007) among newborns born before the 2000-2001 U.S. Environmental Protection Agency's regulatory actions to phase out residential use of these insecticides. Among newborns born after January 2001, exposure levels were substantially lower, and no association with fetal growth was apparent (p > 0.8). The propoxur metabolite 2-isopropoxyphenol in cord plasma was inversely associated with birth length, a finding of borderline significance (p = 0.05) after controlling for chlorpyrifos and diazinon. Results indicate that prenatal chlorpyrifos exposures have impaired fetal growth among this minority cohort and that diazinon exposures may have contributed to the effects. Findings support recent regulatory action to phase out residential uses of the insecticides.
DOI: 10.1289/ehp.6641
Language: English
English
Publication Type: Journal Article
Classification: X 24136 Environmental impact
Classification: M1 110 Population-Environment Relations
Classification: P 6000 TOXICOLOGY AND HEALTH
Classification: H 5000 Pesticides
Subfile: Health & Safety Science Abstracts; Pollution Abstracts; Toxicology Abstracts; Human Population

Wilkinson, F. C. (1979). Dermatophilosis of Sheep Association with Dipping and Effects on Production. Aust.Vet.J. 55: 74-76.

Chem Codes: Chemical of Concern: As,DZ Rejection Code: NO CONC.

Wilkinson, F. C. (1980). The Uptake of Dipping Fluid and Persistence of Diazinon on Shower-Dipped Sheep. Aust.Vet.J. 56: 561-562.

EcoReference No.: 39812

Chemical of Concern: DZ; Habitat: T; Effect Codes: ACC; Rejection Code: NO CONTROL(DZ).

Williams, G. H., Glotfelty, D. E., and Glenn, S. (1986). PESTICIDE WASHOUT IN RAIN. 191st American Chemical Society National Meeting, New York, N.y., Usa, Apr. 13-18, 1986. Abstr Pap Am Chem Soc 191 : No Pagination.

Chem Codes: Chemical of Concern: SZ Rejection Code: FATE.

ABSTRACT: BIOSIS COPYRIGHT: BIOL ABS. RRM ABSTRACT AQUIFER CONTAMINATION BELTSVILLE MARYLAND USA ALACHLOR METACHLOR ATRAZINE SIMAZINE DIAZINON MALATHION BHC HEXACHLOROBENZENE LINDANE CHLORDANE TOXAPHENE

KEYWORDS: General Biology-Symposia
KEYWORDS: Ecology
KEYWORDS: Ecology
KEYWORDS: Biochemical Methods-General
KEYWORDS: Biochemical Studies-General
KEYWORDS: Toxicology-Environmental and Industrial Toxicology
KEYWORDS: Public Health: Environmental Health-Air
KEYWORDS: Agronomy-Weed Control
KEYWORDS: Pest Control
KEYWORDS: Economic Entomology-Chemical and Physical Control

Willson, Michele, Sanchez, Michel, and Labarre, Jean-Francois (1987). The gem-tetraziridinocyclotriphosphazene diazide, N3P3Az4(N3)2, a synthon to monophosphazenylmonoazido derivatives of biological interest. Inorganica Chimica Acta 136: 53-59.

Chem Codes: Chemical of Concern: DZ Rejection Code: METHODS.

This contribution reports on the synthesis of the gem-tetraziridinodiazidocyclotriphosphazene which is a suitable synthon for the synthesis of monophosphazenylmonoazidocyclotriphosphazenes of biological interest.

Wilschut, Jan, Scholma, Janny, Bental, Michal, Hoekstra, Dick, and Nir, Shlomo (1985). Ca2+-induced fusion of phosphatidylserine vesicles: mass action kinetic analysis of membrane lipid mixing and aqueous contents mixing. Biochimica et Biophysica Acta (BBA) - Biomembranes 821: 45-55.
Chem Codes: Chemical of Concern: DZ Rejection Code: METHODS.

We have investigated the initial kinetics of Ca2+-induced aggregation and fusion of phosphatidylserine large unilamellar vesicles at 3, 5 and 10 mM Ca2+ and 15, 25 and 35[deg]C, utilizing the Tb/dipicolinate (Tb/DPA) assay for mixing of aqueous vesicle contents and a resonance energy transfer (RET) assay for mixing of bilayer lipids. Separate rate constants for vesicle aggregation as well as deaggregation and for the fusion reaction itself were determined by analysis of the data in terms of a mass action kinetic model. At 15[deg]C the aggregation rate constants for either assay are the same, indicating that at this temperature all vesicle aggregation events that result in lipid mixing lead to mixing of aqueous contents as well. By contrast, at 35[deg]C the RET aggregation rate constants are higher than the Tb/DPA aggregation rate constants, indicating a significant frequency of reversible vesicle aggregation events that do result in mixing of bilayer lipids, but not in mixing of aqueous vesicle contents. In any conditions, the RET fusion rate constants are considerably higher than the Tb/DPA fusion rate constants, demonstrating the higher tendency of the vesicles, once aggregated, to mix lipids than to mix aqueous contents. This possibly reflects the formation of an intermediate fusion structure. With increasing Ca2+ concentrations the RET and the Tb/DPA fusion rate constants increase in parallel with the respective aggregation rate constants. This suggests that fusion susceptibility is conferred on the vesicles during the process of vesicle aggregation and not solely as a result of the interaction of Ca2+ with isolated vesicles. Aggregation of the vesicles in the presence of Mg2+ produces neither mixing of aqueous vesicle contents nor mixing of bilayer lipids. Membrane fusion/ Phospholipid vesicle/ Liposome/ Phosphatidylserine/ Ca2+

Wilson, Irwin B. and Cohen, Max (1953). The essentiality of acetylcholinesterase in conduction. Biochimica et Biophysica Acta 11: 147-156.
Chem Codes: Chemical of Concern: DZ Rejection Code: METHODS.

A method is described for assaying cholinesterase in an intract crab nerve using acetycholine and dimethylaminoethyl acetate (DMAEA) as substrates. This technique makes possible a new evaluation of the relationship between enzyme activity of nerve fibers and their ability to conduct impulses. It is shown that acethylcholine assays only a portion of the total enzyme, called in this paper external enzyme. DMAEA being a tertiary amine is able to penetrate barriers relatively impermeable to acethylcholine and other quarternary ammonium salts, and to reach extra enzyme called internal enzyme. There is probably about 3 times as much internal as external enzyme.The powerful enzyme inhibitors, prostigmine, tetraethyl pyrophosphate, and decamethonium nearly completely inhibit the external enzyme. DMAEA hydrolysis is inhibited only 50%, the unhibited portion being attributed to internal enzyme. These inhibitors do not produce conduction block. In striking contrast, those enzyme inhibitors which block conduction, such as DFP and eserine, inhibit both internal and external eznyme. It is shown that conduction is blocked by DFP when the total enzyme inhibition reaches about 80%. Complete enzyme inhibition can be obtained, but conduction could not be observed when the hydrolytic activity fell below about 20% (one case 14%). It has not been possible to obtain conduction in the absence of easily measured amounts of enzyme.Under large dosages of X-ray irradiation, but where conduction was not impaired, the internal enzyme activity increased although external enzyme activity declined. This was interpreted as caused by an increased permeability to DMAEA resulting from X-ray exposure.

Wilson, Irwin B. and Ginsburg, Sara (1959). Reactivation of alkylphosphate inhibited acetylcholinesterase by bis quaternary derivatives of 2-PAM and 4-PAM. Biochemical Pharmacology 1: 200-206.
Chem Codes: Chemical of Concern: DZ Rejection Code: IN VITRO.

Symmetrical and unsymmetrical bis-quaternary derivatives of 2-PAM and 4-PAM were synthesized and studied as reactivators in vitro of acetylcholinesterase inhibited by TEPP and DFP. Certain of the 2-PAM-derivatives and all of the 4-PAM-derivatives were far superior to the parent compound in the sense of producing reactivations at lower concentrations. Despite the fact that 2-PAM is much more active than 4-PAM, the bis-quaternary derivatives of the latter were considerably more active in regard to DFP inhibition, but only slightly more active in regard to TEPP inhibition.The best compound with regard to DFP inhibition was the unsymmetrical bisquaternary salt of 4-PAM and pyridine with a pentamethylene linking chain.In general, the addition of extra binding features in these series produced better reactivators.

Wilson, Irwin B. and Sondheimer, Fred (1957). A specific antidote against lethal alkyl phosphate intoxication. V. Antidotal properties. Archives of Biochemistry and Biophysics 69: 468-474.
Chem Codes: Chemical of Concern: DZ Rejection Code: NO COC.

The results of this paper can be summarized by saying that PAM is a very effective antidote in TEPP and sarin poisoning. The success of the antidote is caused by reactivation of the inhibited acetylcholinesterase--the chemical lesion is repaired.Against tabun PAM is much less effective but is still of considerable value. In this case the action depends on limiting the extent of the chemical lesion through reaction directly with free tabun.Practical cases of parathion poisoning properly come under the heading of paraoxon poisoning. Pure parathion is of very low toxicity and had therefore to be used in very large doses. PAM was effective, but for reasons that are discussed, it was not fully evaluated.