A recombinant baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV), has been utilized to express the opd (organophosphate-degrading) gene from Pseudomonas diminuta in insect tissue-culture cells (Sf9) of the fall armyworm (Spodoptera frugiperda). The broad-spectrum organophosphate hydrolase (EC 3.1.8.1) encoded by this gene is a member of a general class of enzymes [organophosphate (OP) anhydrolases] that include parathion hydrolases, di-isopropyl-fluorophosphatases (DFPases), somanases, and OP phosphotriesterases. This particular enzyme possesses the ability to hydrolyse paraoxon (P-O bond), DFP, sarin (P-F bond), VX (P-S bond) and tabun (P-CN bond), as well as a number of other extensively used organophosphorus pesticides. The enzyme produced in infected Sf9 cells is post-translationally processed and resembles the mature form of the enzyme expressed in various bacterial cells as identified by immunoprecipitation on Western blots. N-terminal sequence analysis of enzyme expressed in insect cells revealed Gly-29 as the terminal residue, whereas expression in Escherichia coli removes this residue, exposing Ser-30 at the N-terminus. Conditions for optimal expression of the enzyme in this system are described. Furthermore, hydrolytic efficiency of some OPs with purified enzyme from this system is discussed in relation to the in situ activity of Pseudomonas diminuta MG cells.

Animals↗

PubMed1993

Prevalence of shigellosis and other enteric pathogens in a zoologic collection of primates.

An epidemiologic study of shigellosis was the preliminary step in the formulation of a plan for the control of devastating infectious diseases in nonhuman primates at the National Zoological Park. Data were collected from primate groups with enzootic shigellosis and included the following species: white-cheeked and siamong gibbons (Hylobates concolor and H syndactylies); lion-tailed, celebes, and Barbary macaques (Macaca silenus, M nigera, and M sylvanus); black and white colobus monkeys (Colobus guerzea); grey-cheeked mangabeys (Cerecocebus albigena); spider monkeys (Ateles susciceps robusuts); ruffed lemurs (Lemur varrigatus); lowland gorillas (Gorilla gorilla); and orangutans (pongo pygmaeus). Data included results of physical examination, proctoscopy with biopsy, fecal parasitologic and cytologic examinations, and bacteriologic culturing of swabbed specimens of rectum and gingiva. Repetitive fecal examinations were subsequently performed and included bacteriologic culturing of fecal specimens for enteropathogenic bacteria and parasites and cytologic examination of feces. Data were collected for a 1-year period from 82 primates, and 14 gibbons were studied intensively. White-cheeked and siamang gibbons shed Shigella flexneri sporadically, but persistently. All gibbons were affected with a mean point prevalence of 30.7% (range 0 to 71%). Shigella flexneri also was isolated from feces of lion-tailed macaques. Shigella sonnei was isolated from feces of grey-cheeked mangabeys, celebes macaques, and spider monkeys.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Focus on psychometrics. Development and administration of a survey instrument for cross-cultural research.

The purpose of this article is to provide a brief review of guidelines for development of culturally appropriate measures of phenomena. The principles are illustrated by citing an example of their use in developing a survey instrument to be used with a rural Mexican American and Euro American population. Discussion focuses on practical issues of implementing the approaches suggested in the literature and adds to the literature about issues in formatting and administration of survey instruments used in cross-cultural research.

Cross-Cultural Comparison↗

PubMed1993

An international perspective of active euthanasia: attitudes of nurses in seven countries.

This exploratory study examines the ethical justification that cancer care and dementia care nurses gave for active voluntary euthanasia. A convenient sample of 319 nurses working in seven countries was interviewed using a structured interview guide. The great majority of the nurses could not ethically justify active voluntary euthanasia. Even if the law changed, only 96 of the total sample viewed active voluntary euthanasia as ethical. For those nurses who could ethically justify active voluntary euthanasia, the majority did so because of the patients' suffering.

Adult↗

PubMed1993

Quality of attention in chronic fatigue syndrome: subjective reports of everyday attention and cognitive difficulty, and performance on tasks of focused attention.

Patients with chronic fatigue syndrome (also known as post-viral fatigue syndrome or myalgic encephalomyelitis) commonly report cognitive difficulties concerning attention, concentration and memory. In this study, patients were compared with matched controls on two questionnaires which assess subjective difficulties with attention and general cognitive functioning, and on two tasks requiring focused attention. Patients reported significantly greater difficulty with attention on the Everyday Attention Questionnaire and more cognitive symptoms on the Profile of Fatigue-Related Symptoms. The objective tests did not clearly indicate a deficit in patients' focused attention; patients tended to perform less well on the Embedded Figures Test and the Stroop Colour-Word Interference Test, but these differences were not significant. There was, however, evidence of psychomotor retardation, with patients having longer response times for word reading and colour naming in the Stroop test. Difficulties in interpreting findings for both subjective and objective cognitive measures are discussed.

Adult↗

PubMed1993

Management and treatment of complicated cases of basal cell carcinoma.

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Aged↗

PubMed1992

High-osmolality and low-osmolality contrast agents.

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Arrhythmias, Cardiac↗

PubMed1992

Antifolates induce inhibition of amido phosphoribosyltransferase in leukemia cells.

The pathway for de novo biosynthesis of purine nucleotides contains two one-carbon transfer reactions catalyzed by glycinamide ribotide (GAR) and 5-aminoimidazole-4-carboxamide ribotide (AICAR) transformylases in which N10-formyltetrahydrofolate is the one-carbon donor. We have found that the antifolates methotrexate (MTX) and piritrexim (PTX) completely block the de novo purine pathway in mouse L1210 leukemia cells growing in culture but with only minor accumulations of GAR and AICAR to less than 5% of the polyphosphate derivatives of N-formylglycinamide ribotide (FGAR) which accumulate when the pathway is blocked completely by azaserine. This azaserine-induced accumulation of FGAR polyphosphates is completely abolished by MTX, indicating that inhibition of the pathway is at or before GAR transformylase (reaction 3; Lyons, S. D., and Christopherson, R. I. (1991) Biochem. Int. 24, 187-197). Three h after the addition of MTX (0.1 microM), cellular 5-phosphoribosyl-1-pyrophosphate has accumulated 3.4-fold while 6-methyl-mercaptopurine riboside (25 microM) induces a 6.3-fold accumulation. These data suggest that amido phosphoribosyltransferase catalyzing reaction 1 of the pathway is the primary site of inhibition. In support of this conclusion, we have found that dihydrofolate-Glu5, which accumulates in MTX-treated cells, is a noncompetitive inhibitor of amido phosphoribosyltransferase with a dissociation constant of 3.41 +/- 0.08 microM for interaction with the enzyme-glutamine complex in vitro. Folate-Glu5, MTX-Glu5, PTX, dihydrotriazine benzenesulfonyl fluoride, and AICAR also inhibit amido phosphoribosyltransferase.

Amidophosphoribosyltransferase↗

PubMed1992

Deaths from gastro-intestinal cancer in Mexico: probable cause for water sampling.

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Agriculture↗

PubMed1992

A mathematical model for estimation of broiler egg weight loss from physical dimensions and air cell size during incubation.

A formula was derived that will enable hatcheries to estimate percentage weight loss using the width, length, and air cell diameter of the egg. The formula was derived by fitting an equation to the shape of the egg and using this shape equation to obtain estimates of air cell volume and egg volume. These estimates were coupled with the principle that weight = density x volume to give the final formula as a function of air cell diameter divided by egg width. The formula was tested by comparing its estimated values with measured values from eggs set at two different incubation conditions chosen to cover the range of conditions found in commercial hatcheries. As the day of transfer approached, embryo size and distortion of the air cell caused inaccuracies to develop in the results of this equation. However, data suggest that the equation can be used to calculate percentage weight loss until approximately the 16th day of incubation.

Animals↗

PubMed1992

The distribution of DNA sequences hybridizing with antibiotic production and resistance gene probes within type strains and wild isolates of Streptomyces species.

Total DNA preparations from 74 antibiotic-producing type strains and 102 natural Streptomyces isolates were examined by dot blots for homology to 6 antibiotic production and resistance genes. Pattern diversity of hybridizations decreased as stringency increased from 65% to 85%. There were 146 unique profiles at 65% stringency with 13 repeated patterns, whilst there were only 14 unique and 11 repeated profiles at 85% stringency. Most of the strains which hybridized at 85% reacted with one or two probes although a few strains showed multiple homologies. This data was used to cluster strains and the groups defined were examined for phenotypic antibiotic resistance. Producers of certain classes of antibiotics clustered to specific groups and some gene homologies were more common amongst strains which produced similar antibiotics.

Anti-Bacterial Agents↗

PubMed1991

Amino acid residues in the T cell receptor CDR3 determine the antigenic reactivity patterns of insulin-reactive hybridomas.

We examined TCR gene usage in a panel of beef insulin/I-Ad-restricted T cell hybrids obtained from BALB/c mice. These hybrids demonstrated several distinct patterns of reactivity defined by their ability to respond to species variants of insulin. Correlation of TCR-alpha and -beta-gene usage with these patterns of reactivity demonstrated that TCR gene usage was restricted within Ag reactivity groups. In particular, V-J junctional regions (CDR3 equivalent) were restricted with conserved junctional amino acid motifs present in both TCR-alpha- and -beta-chains. Comparison of TCR gene usage in hybrids expressing identical V alpha and V beta gene segments but demonstrating different patterns of reactivity revealed that changes in either J alpha and/or J beta gene segment usage could alter antigenic reactivity. Indeed, single or limited amino acid differences within the CDR3 region were sufficient to markedly alter fine specificity. These data demonstrate the critical role for CDR3 in determining antigenic reactivity in beef insulin-reactive hybrids and are compatible with the current model of TCR/peptide/MHC interaction.

Amino Acid Sequence↗

PubMed1991

Actin dynamics studied by solid-state NMR spectroscopy.

Solid-state nuclear magnetic resonance spectroscopy was used to study the motion of 2H and 19F probes attached to the skeletal muscle actin residues Cys-10, Lys-61 and Cys-374. The probe resonances were observed in dried and hydrated G-actin, F-actin and F-actin-myosin subfragment-1 complexes. Restricted motion was exhibited by 19F probes attached to Cys-10 and Cys-374 on actin. The dynamics of probes attached to dry cysteine powder or F-actin were very similar and the binding of myosin had little effect indicating that the local probe environment imposes the major influence on motion in the solid state. Correlation times determined for the solid state probes indicated that they were undergoing some rapid internal motion in both G-actin and F-actin such as domain twisting. The probe size influenced the motion in G-actin and appeared to sense monomer rotation but not in F-actin where segmental mobility and intramonomer co-ordination appeared to dominate.

Acetone↗