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Author |
Haruta, N.; Kitagawa, T. |
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Title |
Time-resolved UV resonance Raman investigation of protein folding using a rapid mixer: characterization of kinetic folding intermediates of apomyoglobin |
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Journal Article |
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Year |
2002 |
Publication |
Biochemistry |
Abbreviated Journal |
Biochemistry |
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Volume |
41 |
Issue |
21 |
Pages |
6595-6604 |
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Keywords |
Animals; Apoproteins/*chemistry; Circular Dichroism; Holoenzymes/chemistry; Horses; Hydrochloric Acid/chemistry; Hydrogen-Ion Concentration; Imidazoles/chemistry; Kinetics; Models, Molecular; Myoglobin/*chemistry; Peptide Fragments/chemistry; *Protein Folding; Protein Structure, Secondary; Spectrum Analysis, Raman/*methods; Tryptophan/*chemistry; Ultraviolet Rays; Whales |
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Abstract |
The 244-nm excited transient UV resonance Raman spectra are observed for the refolding intermediates of horse apomyoglobin (h-apoMb) with a newly constructed mixed flow cell system, and the results are interpreted on the basis of the spectra observed for the equilibrium acid unfolding of the same protein. The dead time of mixing, which was determined with the appearance of UV Raman bands of imidazolium upon mixing of imidazole with acid, was 150 micros under the flow rate that was adopted. The pH-jump experiments of h-apoMb from pH 2.2 to 5.6 conducted with this device demonstrated the presence of three folding intermediates. On the basis of the analysis of W3 and W7 bands of Trp7 and Trp14, the first intermediate, formed before 250 micros, involved incorporation of Trp14 into the alpha-helix from a random coil. The frequency shift of the W3 band of Trp14 observed for this process was reproduced with a model peptide of the A helix when it forms the alpha-helix. In the second intermediate, formed around 1 ms after the start of refolding, the surroundings of both Trp7 and Trp14 were significantly hydrophobic, suggesting the formation of the hydrophobic core. In the third intermediate appearing around 3 ms, the hydrophobicity was relaxed to the same level as that of the pH 4 equilibrium intermediate, which was investigated in detail with the stationary state technique. The change from the third intermediate to the native state needs more time than 40 ms, while the appearance of the native spectrum after the mixing of the same solutions was confirmed separately. |
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School of Mathematical and Physical Sciences, The Graduate University for Advanced Studies, Myodaiji, Okazaki 444-8585, Japan |
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0006-2960 |
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PMID:12022863 |
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Equine Behaviour @ team @ |
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3785 |
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Author |
Voss, B.; Mohr, E.; Krzywanek, H. |
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Title |
Effects of aqua-treadmill exercise on selected blood parameters and on heart-rate variability of horses |
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Journal Article |
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Year |
2002 |
Publication |
Journal of Veterinary Medicine. A, Physiology, Pathology, Clinical Medicine |
Abbreviated Journal |
J Vet Med A Physiol Pathol Clin Med |
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Volume |
49 |
Issue |
3 |
Pages |
137-143 |
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Keywords |
Animals; Electrocardiography/veterinary; Exercise Test/veterinary; Female; Heart Rate/*physiology; Hemoglobins/metabolism; Horses/*physiology; Lactic Acid/blood; Male; Physical Conditioning, Animal/*physiology; Water |
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The objectives of the present study were to investigate the effects of Aquatraining of horses (aqua-treadmill exercise; treadmill manufactured by Equitech – L.u.S. Equipment, Warendorf, Germany) on selected blood parameters [lactic acid concentration (mmol/l), haemoglobin content (g/l)] and on heart-rate variability (HRV) [heart rate (beats per min; b.p.m.), standard deviation of all NN-intervals (SDNN; ms), normalized power of the low and high frequency band (LFnorm, Hfnorm; au), % recurrence, % determinism and ratio(corr)]. Seven horses performed six exercise tests with different work loads (walking (x = 1.56 +/- 0.08 m/s) and trotting (x = 2.9 +/- 0.13 m/s): dry, water above the carpus and water above the elbow). The standardized test-protocol was: 5 min warm-up at walk while the water was pumped in, followed by the 20-min exercise period at walk or trot, followed by a 5-min walk while pumping out the water. Blood samples were taken prior to each test at rest in the stable, as well as exactly 5 min after the end of the 20-min exercise period. Electrocardiograms were recorded during rest and the 20-min exercise period. Compared to rest, neither the chosen velocities, the two water levels, nor the dry tests led to a significant increase of the lactic acid concentration in any horse. The haemoglobin content showed a significant increase as a result of exercise. Significant differences could be found between the heart rates at rest and the six exercise tests and between the mean of the levels 'walking' and the mean of the levels 'trotting'. An exercise-induced change of HRV was characterized by a decreasing SDNN, a significantly higher LFnorm (sympathetic influence) combined with a significantly lower HF(norm) power (parasympathetic activity) and a rising degree of order (significantly higher % determinism and nearly unchanged % recurrence) and stability (significantly rising ratio(corr)) of the recurrence plot. In conclusion, the used training-protocol for aqua-treadmill exercises only represents a medium-sized aerobic work load for horses, but the different levels of burden were indicated especially by changes in HRV. |
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Institute for Veterinary Physiology of the Free University Berlin, Germany |
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0931-184X |
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PMID:12019954 |
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Call Number |
Equine Behaviour @ team @ |
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4049 |
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Author |
Griffin, B. |
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Title |
The use of fecal markers to facilitate sample collection in group-housed cats |
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Journal Article |
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Year |
2002 |
Publication |
Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science |
Abbreviated Journal |
Contemp Top Lab Anim Sci |
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41 |
Issue |
2 |
Pages |
51-56 |
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Keywords |
Animals; Behavior, Animal; Biological Markers/*analysis; Cats/*physiology/psychology; Diet/veterinary; Feces/*chemistry; Food Coloring Agents/analysis; Housing, Animal; Individuality; Plastics/analysis; Specimen Handling/methods/*veterinary |
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The provision of proper social housing is a priority when designing an experiment using domestic cats as laboratory animals. When animals are group-housed, studies requiring analysis of stool samples from individual subjects pose difficulty in sample collection and identification. In this study, commercially available concentrated food colorings (known as bakers pastes) were used as fecal markers in group-housed cats. Cats readily consumed 0.5 ml of bakers paste food coloring once daily in canned cat food. Colorings served as fecal markers by imparting a distinct color to each cat s feces, allowing identification in the litter box. In addition, colored glitter (1/8 teaspoon in canned food) was fed to cats and found to be a reliable fecal marker. Long-term feeding of colorings and glitter was found to be safe and effective at yielding readily identifiable stools. |
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Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University, Alabama 36841, USA |
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1060-0558 |
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PMID:11958604 |
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Equine Behaviour @ team @ |
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4165 |
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Chamove, A.S.; Crawley-Hartrick, O.J.E.; Stafford, K.J. |
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Title |
Horse reactions to human attitudes and behavior |
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Year |
2002 |
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Anthrozoos |
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15 |
Issue |
4 |
Pages |
323-331 |
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Cited By (since 1996): 4; Export Date: 21 October 2008 |
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Equine Behaviour @ team @ |
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4514 |
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Author |
Ginther, O.J.; Lara, A.; Leoni, M.; Bergfelt, D.R. |
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Title |
Herding and snaking by the harem stallion in domestic herds |
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2002 |
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Theriogenology |
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57 |
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8 |
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2139-2146 |
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Cited By (since 1996): 1; Export Date: 21 October 2008 |
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Equine Behaviour @ team @ |
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4516 |
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Jedrzejewski, W.; Schmidt, K.; Theuerkauf, J.; Jedrzejewska, B.; Selva, N.; Zub, K. |
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Kill rate and predation by wolves on ungulate populations in Bialowieza primeval forest (Poland) |
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2002 |
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Ecology |
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83 |
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Equine Behaviour @ team @ Jedrzejewski2002 |
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6481 |
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