Gene diversity at allozyme loci in the cottonwood leaf beetle,Chrysomela scripta

Gene diversity was studied in beetles collected from a poplar plantation at Ames, Iowa. The beetles are widely distributed throughout North America wherever poplars (Populus spp) occur. Of 38 loci interpretable by simple Mendelian criteria, 22 were polymorphic (58%). Nei's mean heterozygosity per locus was 20.1±4.0%. A mean 2.29±1.35 alleles per locus was detected. The foregoing levels of diversity are typical of Coleoptera. There were significant deviations from Hardy-Weinberg expectations at six loci, and of these loci, four deviations were judged to have been caused by technical problems in resolving heterozygotes. Twelve polymorphic loci are considered suitable to carry out studies on the breeding structure of this important pest.     

The activity of mammalian peroxidases (lactoperoxidase and myeloperoxidase) and their compounds III toward 2-t-butyl-4-methoxyphenol (butylated hydroxyanisole) and its dimer (2,2'-dihydroxy-3,3'-di-t-butyl-5,5'-dimethoxydiphenyl).

Spectral evidence is presented which shows that butylated hydroxyanisole (BHA) and its dimer act as electron donors for lactoperoxidase (LPO) and myeloperoxidase (MPO) by two different pathways: peroxidative and oxidative. LPO compound II and MPO compound II are converted to native enzymes in their reactions with BHA without detectable intermediates. This confirms a normal peroxidatic oxidation of this commonly used antioxidant. We also report spectral data indicating the reductions of peroxidase compound III to the native state in reactions with BHA (LPO, MPO) or with di-BHA (LPO). This oxidative reaction has significant physiological relevance, ensuring return of peroxidases to the native state for re-entry into the normal peroxidatic cycle or into halogenating reactions.     

Cyclopentenylcytosine triphosphate. Formation and inhibition of CTP synthetase

Cyclopentenylcytosine (CPEC) is phosphorylated in L1210 cells with CPEC triphosphate as the major metabolite. Partially purified uridine-cytidine kinase catalyzes the initial phosphorylation of cyclopentenylcytosine with an apparent Km of 196 +/- 9 microM, and cyclopentenylcytosine is a competitive inhibitor of cytidine phosphorylation by this enzyme with a Ki value of 144 +/- 14 microM. Examination of the CTP synthetase activity in extracts of L1210 cells revealed a dose-dependent decrease on exposure of cells to CPEC. Synthesis of CPEC triphosphate by an enzymatic method permitted direct examination of the inhibition of partially purified CTP synthetase. CPEC triphosphate inhibited bovine CTP synthetase with a median inhibitory concentration of 6 microM, whereas CPEC mono- and diphosphates were ineffective. CTP synthetase showed a classical Michaelis-Menten hyperbolic plot of velocity and UTP concentration in the presence of saturating concentrations of ATP and glutamine, but CPEC triphosphate induced sigmoidal kinetic plots. The Hill coefficient was calculated to be 3.2.     

Binding of concanavalin A by normal, herpes virus-transformed, and trypsin-treated hamster embryo fibroblasts

Using fluorescein isothiocyanate-labeled concanavalin A (ConA), it was shown that normal hamster embryo fibroblast cells appear to bind less ConA than do herpes virus-transformed cells. However, the binding capacity of normal HEF cells could be increased following trypsin treatment.     

Evaluating automated infrared thermography and vulva exposure tracking as components of an estrus detection platform in a commercial dairy herd

The primary objective of this study was to develop an automated infrared thermography platform (Estrus BenchMark) capable of measuring skin temperature and tail movements as a means of identifying cows in estrus. The secondary objective was to evaluate the accuracy of Estrus BenchMark to detect estrus compared to in-line milk progesterone (P₄) analysis (Herd Navigator System) in a commercial dairy herd managed under a robotic milking system. Data were collected on forty-six cows from 45 to 120 d after calving. Cows were flagged in estrus when milk P₄ fell below 5 ng/mL. The Estrus BenchMark true positive estrus alerts (Sensitivity; Se%) were compared to Herd Navigator System estrus alerts at different time-windows (±12 h, ±24 h, ±48 h, and ±72 h) relative to the Estrus BenchMark estrus alerts for all the estrus alerts (AE) and confidence-quality estrus (CQE; >80% quality) alerts identified by Herd Navigator System. The Estrus BenchMark captured skin temperature and tail movements resulting in vulva exposure (left tail movements, LTail; right tail movements, RTail; and pooled tail movements, PTail) for each milking event. Skin temperature tended to increase when the milk P₄ concentration (Least-Squares Means ± SE) dropped for AE (estrus day [d 0]; P₄; 3.51 ± 0.05 ng/mL, Skin temperature; 33.31 ± 2.38 °C) compared with d ?7 (P₄; 20.22 ± 0.73 ng/mL; Skin temperature: 32.05 ± 3.77 °C). The increase in skin temperature, however, was significant in cows with CQE > 80% at d 0 (32.75 ± 0.29 °C) compared to d ?7 (31.80 ± 0.28 °C). The prevalence of tail movements to expose vulva was greater (P = 0.01) in AE at d 0 (LTail: 62.50%; PTail; 68.75%; and RTail: 56.25%) compared with d ?7 (LTail: 18.75%; PTail: 9.37%: and RTail: 9.37%), and d +4 (LTail: 9.37%; PTail: 9.37%; and RTail: 12.5%). Moreover, the higher prevalence of tail movements at d 0 was observed in cows with CQE > 80% (LTail; 65%, PTail; 80%, and RTail; 70%) compared to those with CQE < 80%. The highest Estrus BenchMark Youden index (YJ; 0.45), diagnostic odds ratio (DOR; 9.04), and Efficiency (0.77) were achieved for AE in a ±48 h window and at ±72 h window for CQE (YJ; 0.66, DOR; 25.29, and Efficiency 0.76) relative to Herd Navigator System estrus alerts. The highest Estrus BenchMark resulted in 58% estrus detection rates for AE and 80% for cows with CQE compared to the Herd Navigator System.     

Effect of DNA modifications on DNA processing by HIV-1 integrase and inhibitor binding: role of DNA backbone flexibility and an open catalytic site

Integration of the viral cDNA into host chromosomes is required for viral replication. Human immunodeficiency virus integrase catalyzes two sequential reactions, 3'-processing (3'-P) and strand transfer (ST). The first integrase inhibitors are undergoing clinical trial, but interactions of inhibitors with integrase and DNA are not well understood in the absence of a co-crystal structure. To increase our understanding of integrase interactions with DNA, we examined integrase catalysis with oligonucleotides containing DNA backbone, base, and groove modifications placed at unique positions surrounding the 3'-processing site. 3'-Processing was blocked with substrates containing constrained sugars and alpha-anomeric residues, suggesting that integrase requires flexibility of the phosphodiester backbone at the 3'-P site. Of several benzo[a]pyrene 7,8-diol 9,10-epoxide (BaP DE) adducts tested, only the adduct in the minor groove at the 3'-P site inhibited 3'-P, suggesting the importance of the minor groove contacts for 3'-P. ST occurred in the presence of bulky BaP DE DNA adducts attached to the end of the viral DNA suggesting opening of the active site for ST. Position-specific effects of these BaP DE DNA adducts were found for inhibition of integrase by diketo acids. Together, these results demonstrate the importance of DNA structure and specific contacts with the viral DNA processing site for inhibition by integrase inhibitors.     

New Polymorphic Microsatellites in Glossina pallidipes (Diptera: Glossinidae) and Their Cross-Amplification in Other Tsetse Fly Taxa

We report the development and characterization of three new microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae). Fifty-eight alleles were scored in 192 individuals representing six natural populations. Allelic diversity ranged from 9 to 28 alleles per locus (mean 19.3 +/- 5.5). Averaged across loci, observed heterozygosity was 0.581 +/- 0.209, and expected heterozygosity was 0.619 +/- 0.181. Cross-species amplifications of the G. pallidipes loci in other tsetse fly taxa are reported.     

Skeletal muscle fiber composition of the English sparrow (Passer domesticus)

Substrate utilization by English sparrow skeletal muscle has been extensively studied in our lab. However, there are few published studies on the muscle fiber composition of English sparrow wing and gastrocnemius muscles. The objective of the present study was to examine the fiber type composition of a variety of muscles in the English sparrow. The classification of a muscle fiber as fast glycolytic, slow oxidative, or fast oxidative glycolytic provides insight into the physiological function of muscles. Therefore, we completed mATPase and NADH stains on four muscles of the sparrow wing, as well as the gastrocnemius muscle, to characterize these muscle fiber types. Results show that the fibers of extensor digitorum communis, extensor metacarpi ulnaris, and extensor metacarpi radialis are homogeneous fast oxidative. The fibers of the supinator are homogeneous fast oxidative in 62.5% of samples, and heterogeneous (45.2% fast oxidative, 54.8% fast nonoxidative) in 37.5% of samples. Whereas the gastrocnemius muscle fibers are heterogeneous (10% fast oxidative, 64% fast nonoxidative, 26% slow oxidative) in all muscles examined.