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PubMed1989

Spinal hyperostosis in comparative pathology. A useful approach to the concept.

Spinal hyperostosis, an anatomical and radiological concept primarily described in man, is characterized by enthesopathic bony overgrowth on vertebral bodies in the form of spurs or intervertebral bridges. It can also be part of a more diffuse enthesopathic condition, including the appendicular skeleton. These changes are distinct from those of osteoarthrosis. Similar changes can be observed in all kinds of mammals, independent of their type of locomotion (bipodic, quadrumanous, quadrupedic, or aquatic). An anatomical and radiological study is presented of six cases (with histological examination of two dogs and one horse, and observation of macerated specimens of one horse, one equida, and one whale). Comparative pathology allows for a better understanding of the concept of hyperostosis and, even more clearly than in man, this can be distinguished from that of discovertebral pathology although both conditions could be associated in older subjects. The knowledge of spinal hyperostosis in veterinary medicine could--as in man--be of interest to differentiate between this process and other features of vertebral pathology.

Animals↗
PubMed1992

Effects of amino acid substitutions outside an antigenic site on protein binding to monoclonal antibodies of predetermined specificity obtained by peptide immunization: demonstration with region 145-151 (antigenic site 5) of myoglobin.

Monoclonal antibodies of predetermined specificity were prepared by immunization with a free (i.e., without coupling to any protein carrier) synthetic peptide representing region 145-151 of sperm whale myoglobin (SpMb) and their cross-reactions with eight Mb variants were determined. Five Mbs--bottle-nose dolphin myoglobin (BdMb), pacific common dolphin myoglobin (PdMb), horse myoglobin (HsMb), dog myoglobin (DgMb), and badger myoglobin (BgMb)--have an identical sequence in that region. Nevertheless, these Mbs exhibited very different cross-reactivities. BdMb and PdMb exhibited cross-activities which were comparable to that of the reference antigen, SpMb; while the reactivity of HsMb was remarkedly decreased, DgMb and BgMb showed almost no cross-reactions with these mAbs. Since the region 145-151 has an identical sequence in all the five Mbs, it is concluded that the differences in their antigenic reactivities with anti-region 145-151 mAbs are due to the effects of amino acid substitutions outside the region 145-151. Another pair of myoglobins, echidna myoglobin (EdMb) and chicken myoglobin (ChMb), have the same sequence in that region, but reacted very differently with anti-region 145-151 mAbs. The reactivity and affinity of EdMb were substantially decreased while those of ChMb were almost completely absent, relative to SpMb. It is concluded, contrary to popular assumptions, that when an amino acid substitution influences the binding of a protein variant to a mAb, it is not necessary for that substitution to be an actual contact residue (i.e., a residue within the antigenic site where the mAb binds). Such effects, which are often very drastic, could be due to indirect influences of the substitution on the chemical and binding properties of the site residues. Furthermore, residues which had been postulated, on the basis of these assumptions, to constitute discontinuous antigenic sites in SpMb, were found [from the present studies and those recently reported with mAbs against the other four antigenic site of Mb (regions 15-22, 56-62, 94-100, and 113-120 of SpMb)] to merely be exerting indirect effects on the known five antigenic sites of Mb. The effects of substitutions, which can happen even in the absence of conformational changes, are determined by many factors, such as the chemical nature of the substitution, its environment, its distance from the site, and the nature of the site residue(s) being affected.

Amino Acid Sequence↗
PubMed1977

On urea formation in marine mammals.

Ornithine carbamoyltransferase (EC 2.1.3.3) has been determined in homogenates of liver of the sei whale (Balaenoptera borealis), the bottle-nose dolphin (porpoise) (Tursiops truncatus) and California sea lion (Zalophus californianus). These marine mammals show levels of this ornithine-urea cycle enzyme which are typical of terrestrial mammals.

Animals↗
PubMed1979

Occurrence of plant sterols in aquatic vertebrates.

Plant sterols were found by gas liquid chromatography in the sterols of five species of aquatic vertebrates; mackerel (Scomber japonicus), rainbow trout (Salmo gairdnerii), smelt (Osmerus dentex), sardine (Sardinops melanosticta) and chimera (Chimera phantasma). The sterols of chimera liver, sardine flesh and sardine viscera contained 9.0, 2.4 and 3.1% of C28 and C29 sterols in addition to 86.7, 96.6 and 95.2% of cholesterol. The occurrence of norcholestandienol, campesterol, beta-sitosterol and C28 stanol was shown by combined gas chromatography-mass spectrometry. Sperm whale (Physeter catodon) sterols consisted of more than 99% cholesterol with only traces of 22-dehydrocholesterol.

Animals↗
PubMed2000

"Spring rise" of whaleworm (Anisakis simplex; Nematoda, Ascaridoidea) third-stage larvae in some fish species from Norwegian waters.

The seasonal variation in the infection of saithe (Pollachius virens), cod (Gadus morhua), and redfish (Sebastes marinus) from a coastal area of central Norway with Anisakis simplex third-stage larvae (L3) was studied over a period of 1 year. In all three host species there was an increase in the abundance of the parasite in spring, with a peak appearing in March and April. Cod displayed the most distinct seasonal variation, showing a clear abundance peak in April. The abundance peak in April for redfish was not as pronounced. In saithe the abundance over the seasons was less prominent, with a maximum appearing in March and another increase in abundance occurring during July. It is apparent that the causes behind the pattern of infection observed in this study are complex. However, possible explanatory mechanisms are discussed. Arguments are propounded to suggest that the increased supply of parasite eggs from northward-migrating whales in addition to the general spring bloom of plankton constitute the most important factors governing the phenomenon of "spring rise" in A. simplex L3 in the study area.

Animals↗
PubMed2004

Polybrominated diphenyl ether congeners and toxaphene in selected marine standard reference materials.

Polybrominated diphenyl ether (PBDE) congeners and components of the complex mixture toxaphene are stable in the environment and readily bioaccumulated into wildlife and human tissues. PBDEs are presently used in large quantities worldwide as flame retardants in textiles, furniture, computer equipment, and cables. Toxaphene is a complex mixture of chlorinated bornanes and bornenes that was the most heavily used pesticide in the United States until it was banned in 1982; however, some countries continue to use toxaphene. The National Institute of Standards and Technology has quantified PBDE congeners and toxaphene in several available Standard Reference Materials (SRMs) using methods of gas chromatography with electron impact mass spectrometry (GC-EI-MS) and GC negative chemical ionization (NCI) MS, respectively. SRM 1588a Organics in Cod Liver Oil and SRM 1945 Organics in Whale Blubber were examined for PBDE congeners 47, 99, 100, 153, and 154, total toxaphene, and toxaphene congeners 26, 50, and 62. SRM 1946 Lake Superior Fish Tissue was also examined for total toxaphene and toxaphene congeners. The sum of the PBDE congeners (mean, (1 SD) wet basis) for SRM 1945 was 150 ng g(-1) (7 ng g(-1)). The concentration of PBDE 47 in SRM 1588a was 82.7 ng g(-1) (2.8 ng g(-1)). Other PBDEs were detected in SRM 1588a but were not quantified due their low levels. The total toxaphene (wet mass basis) was 1,210 ng g(-1) (127 ng g(-1)), 1,960 ng g(-1) (133 ng g(-1)), and 3,980 ng g(-1) (248 ng g(-1)) in SRMs 1945, 1946, and 1588a, respectively. The values for PBDEs and toxaphene determined in the SRMs, while not certified, indicate that the SRMs will be suitable control materials for PBDE and toxaphene analyses.

Adipose Tissue↗
PubMed2000

Magnetic resonance imaging measurements of bone density and cross-sectional geometry.

Magnetic resonance imaging (MRI) is commonly used in the assessment of the musculoskeletal system and associated pathology. The ability of MRI to measure the signals from water and lipid protons enables quantitative measurements of bone porosity. The goal of this investigation was to demonstrate that the density and cross-sectional geometry of whole bones can be noninvasively measured using MRI. Ten trabecular specimens cored from whale vertebrae were used to compare apparent bone density measured directly, and using a quantitative MRI algorithm. Bone density and several cross-sectional geometric properties were also measured using MRI in the distal tibia of 14 volunteers. The MRI measurements were compared with measurements made using quantitative-computed tomography (QCT). A proton density sequence was used for all MRI studies. A porosity phantom was included in the MRI examinations and used to convert the MRI signal intensity to bone volume fraction. Bone density and cross-sectional bone geometry were calculated from the bone volume fractions by assuming constant tissue properties. The apparent density of trabecular bone cores measured directly and using quantitative MRI were linearly related (r(2) = 0.959; P < 0. 01). A strong linear relation also existed between MRI and QCT measurements of ash density (r(2) = 0.923; P < 0.01) and cross-sectional geometric properties (r(2) = 0.976-0.992; P < 0.01). MRI data can be used to measure bone density and cross-sectional geometry of whole bones if a proton density sequence is used to homogenize differences in marrow composition and a porosity phantom is used for slice-specific volume fraction calibration.

Adult↗
PubMed2002

Global distribution of halogenated dimethyl bipyrroles in marine mammal blubber.

Four halogenated dimethyl bipyrroles (HDBPs), hypothesized to be naturally produced, were quantitated in marine mammal blubber from a number of species obtained from various locations worldwide. HDBPs were found in samples from all locations studied. Concentrations of total HDBPs (SigmaHDBPs) ranged from 0.4 ng/g lipid weight in ringed seals (Phoca hispida) from the White Sea to 2,540 ng/g lipid weight in Dall's porpoise (Phocoenoides dalli) from the northwestern North Pacific Ocean. At their highest levels, SigmaHDBPs made up 11% of the total quantitated organohalogen body burden of adult male Dall's porpoises. In two beluga (Delphinapterus leucas) data subsets, it was found that males contained significantly higher concentrations of SigmaHDBPs than females. No significant effects of age or sex on SigmaHDBPs were observed in harbor seal (Phoca vitulina) and bowhead whale (Balaena mysticetus) data subsets. The geographical distribution of concentrations did not resemble that of the ubiquitous anthropogenic organohalogen, polychlorinated biphenyl congener CB-153. Higher concentrations of HDBPs and different patterns of congeners were observed in samples from Pacific as opposed to non-Pacific Ocean influenced environments. Concentrations of HDBPs in beluga from the Arctic and St. Lawrence River were similar. Their high abundance in north Pacific Ocean biota and widespread occurrence suggest that HDBPs undergo extensive transport from a source located primarily in the Pacific Ocean. Evidence from HDBP congener patterns indicates that both ocean currents and atmospheric transport likely play a role in the movement of HDBPs. These results imply that HDBPs and anthropogenic organohalogens have different sources and support the natural production hypothesis.

Adipose Tissue↗
PubMed1998

Methyl sulphone metabolites of polychlorinated biphenyls (PCBs) in cetaceans from the Irish and the Aegean Seas.

The levels of PCBs and PCB methyl sulphone metabolites were determined in the blubber of six species of cetaceans that originated from the Irish Sea and the Aegean Sea (Mediterranean). Burdens ranging from 2.80 to 27.80 microg g-1 (lipid weight) total PCBs and 0.03 to 0.58 microg g-1 (lipid weight) total methyl sulphones were recorded. The highest cetacean methyl sulphone burden (0.58 microg g-1) was in an Irish Sea harbor porpoise (Phocoena phocoena). Methyl sulphone isomer formation capacity, as determined by PCB:MSF ratios, varied considerably between species as follows: harbour porpoise (1:10) > pilot whale (Globicephalus melas), white-sided dolphin (Lagenorhyncus acutus) (1:50) > common dolphin (Delphinus delphis), Risso's dolphin (Grampus griseus), and striped dolphin (Stenella coerueoalba), (1:100), and was related to interspecific variation in PCB methyl sulphone formation capacity.

Adipose Tissue↗
PubMed2002

A water network within a protein: temperature-dependent water ligation in H64V-metmyoglobin and relaxation to deoxymyoglobin.

The sperm whale myoglobin mutant H64V, where the distal histidine is mutated to valine, is known to be five coordinated in the ferric state at room temperature and physiological pH. A change of the ligation in this H64V-Mbmet has been observed by optical absorption spectroscopy as a function of temperature from 20 K to 300 K. Above the dynamical transition at about 180 K one observes the temperature-dependent equilibrium between five- and six-ligated heme. Below the dynamical transition the equilibrium is frozen-in at about 50% of six-coordinate molecules. The water ligation of the iron occurs at temperatures where protein-specific motions are present, as monitored by Mössbauer spectroscopy. The X-ray structures of H64V-Mbmet at 300 K and 110 K are reported with a resolution of 1.5 A and 1.3 A, respectively. The measurements at high resolutions are possible owing to crystallization in the space group P2(1), whereas all mutant myoglobins studies up to now have been carried out with crystals in the space group P6. The overall structure at both temperatures is very close to the native myoglobin. The binding of water at the sixth coordination site at lower temperatures is possible owing to a stabilizing water network extending from the protein surface to the active centre. The reduction of the H64V-Mbmet by electrons obtained by X-ray irradiation of the water-glycerol solvent at 85 K produces an intermediate low-spin state of the water-ligated molecules where Fe(II) retains the six-fold coordination. Mössbauer spectroscopy shows that the relaxation of the metastable low-spin state to high-spin H64V-Mbdeoxy with dissociation of the Fe(II)-H(2)O bond starts at about 115 K and is completed at about 170 K. Differences in the dynamics properties of the native and mutant myoglobin and the connection to the dynamical transition around 180 K are discussed.

Animals↗
PubMed1998

Near-ultraviolet circular dichroic activity of apomyoglobin: resolution of the individual tryptophanyl contributions by site-directed mutagenesis.

The individual tryptophanyl contributions to the near-ultraviolet circular dichroic activity of apomyoglobin in its native conformation have been resolved by studying recombinant proteins with single tryptophanyl substitutions. Site-directed mutagenesis of sperm whale apomyoglobin was performed in order to obtain proteins containing only Trp A-5 or Trp A-12. These amino acid substitutions have very little effect on the overall globin fold as indicated by comparing the spectroscopic properties of the mutants with those of the wild type protein. The circular dichroism spectra of the two apomyoglobin mutants in the near ultraviolet were found to be significantly different, both indole residues having significant activity but of opposite sign. In particular, Trp A-5 shows the presence of a main positive peak centered near 294-295 nm with a marked shoulder at 285 nm, ascribed to the 1LB transition. The spectrum of the mutant protein containing only Trp A-12 shows a large negative contribution with a minimum near 283 nm and a marked shoulder at 293 nm. The broadness of the negative contribution exhibited by Trp A-12 suggests that it may originate mainly from the 1LA transition.

Animals↗
PubMed1998

From metmyoglobin to deoxy myoglobin: relaxations of an intermediate state.

Metmyoglobin has been reduced at low temperature (below 100 K) using x-rays or by excitation of tris(2,2,bipyridine)ruthenium(II) chloride with visible light. Upon reduction, an intermediate state is formed where the structure of the protein is very similar to that of metmyoglobin with the water molecule still bound to the heme iron, but the iron is II low spin. The nature of the intermediate state has been investigated with optical spectroscopy. The Q(O) and Q(V) bands of the intermediate state are split, suggesting that the protoporphyrin is distorted. The intermediate state undergoes a relaxation observed by a shifting of the Soret band at temperatures above 80 K. Above 140 K, the protein begins to relax to the deoxy conformation. The relaxation kinetics of the protein have been monitored optically as a function of time and temperature from minutes to several hours and from 150 K to 190 K. By measuring the entire visible spectrum, we are able to distinguish between electron transfer processes and the protein relaxation from the intermediate state to deoxy myoglobin. The relaxation has been measured in both horse myoglobin and sperm whale myoglobin with the relaxation occurring on faster time scales in horse myoglobin. Both the reduction kinetics and the relaxation show non-exponential behavior. The reduction kinetics can be fit well to a stretched exponential. The structural relaxation from the intermediate state to the deoxy conformation shows a more complex, dynamical behavior and the reaction is most likely affected by the relaxation of the protein within the intermediate state.

2,2'-Dipyridyl↗
PubMed2003

High concentrations of isovaleric acid in the fats of odontocetes: variation and patterns of accumulation in blubber vs. stability in the melon.

Isovaleric acid (iso5:0) is an unusual fatty acid that is important for echolocation and hearing in acoustic tissues of some odontocetes, but its functional significance in blubber is unknown. We examined patterns of accumulation of this compound in blubber in 30 species of odontocetes ( n=299). Iso5:0 concentrations in blubber varied with phylogeny, ontogeny and body topography. Iso5:0 accumulated in greater quantities in superficial/outer blubber than in deep/inner blubber. In the outer blubber of northern right whale and Hector's dolphins, iso5:0 accounted for one-third to one-half of all fatty acids. Total blubber burden of iso5:0 in harbour porpoises represented up to 15 times the amount deposited in the melon. The composition of the melon does not change during starvation in harbour porpoises, supporting the hypothesis that lipids in melon are conserved for a specific function. Some odontocetes continually deposit iso5:0 in blubber after levels in melon have reached asymptotic levels, suggesting independent control of iso5:0 synthesis and storage in these compartments. Dolphins and porpoises inhabiting cold waters possess higher concentrations of iso5:0 in their outer blubber layers than species from warmer regions. We propose that this relationship represents an adaptive secondary role for iso5:0 in maintaining blubber flexibility in cold environments.

Acclimatization↗
PubMed2004

Catalytic activity, stability, unfolding, and degradation pathways of engineered and reconstituted myoglobins.

The structural and functional consequences of engineering a positively charged Lys residue and replacing the natural heme with a heme-L-His derivative in the active site of sperm whale myoglobin (Mb) have been investigated. The main structural change caused by the distal T67K mutation appears to be mobilization of the propionate-7 group. Reconstitution of wild-type and T67K Mb with heme-L-His relaxes the protein fragment around the heme because it involves the loss of the interaction of one of the propionate groups which stabilize heme binding to the protein. This modification increases the accessibility of exogenous ligands or substrates to the active site. The catalytic activity of the reconstituted proteins in peroxidase-type reactions is thus significantly increased, particularly with T67K Mb. The T67K mutation slightly reduces the thermodynamic stability and the chemical stability of Mb during catalysis, but somewhat more marked effects are observed by cofactor reconstitution. Hydrogen peroxide, in fact, induces pseudo-peroxidase activity but also promotes oxidative damage of the protein. The mechanism of protein degradation involves two pathways, which depend on the evolution of radical species generated on protein residues by the Mb active species and on the reactivity of phenoxy radicals produced during turnover. Both protein oligomers and heme-protein cross-links have been detected upon inactivation.

Animals↗
PubMed1985

Continuous gas-depletion technique for measuring O2-dissociation curves of high-affinity hemoglobins.

The O2-depletion technique allows a continuous measurement of the complete O2-dissociation curve of high-affinity hemoglobins in the O2-partial pressure range of 700 to 10(-4) mm Hg. With this technique p50 values greater than or equal to 0.01 mm Hg (approximately equal to 1.3 Pa) can be determined for 3-microliter hemoglobin samples. As examples, O2-dissociation curves of sperm whale myoglobin (p50 = 0.9 mm Hg, 25 degrees C, pH 8.0) and Dicrocoelium hemoglobin (p50 = 0.06 mm Hg, 25 degrees C, pH 4.9) are demonstrated which show Hill coefficients of n = 1.0 over the complete O2-saturation range.

Animals↗
PubMed1986

Myoglobin interspecies structural differences: effects on autoxidation and oxygenation.

Thermodynamic and kinetic parameters of the oxygenation and autoxidation reactions were determined for yellowfin tuna (Thunnus albacares), Pacific green sea turtle (Chelonia mydas caranigra), and sperm whale myoglobin. These proteins have quite similar heme contact residues, but vary in residues which are involved in globin stabilization and in postulated ligand paths to the heme iron. Oxygen equilibria measurements from 10 to 40 degrees C reveal that green sea turtle myoglobin has a lower oxygen affinity and lower enthalpy and entropy for oxygen-binding than the other proteins. Yellowfin tuna myoglobin has the most rapid oxygen dissociation rate and the highest susceptibility to autoxidation under all conditions studied. However, the dependence of the autoxidation rate on ionic strength is the same for the three proteins, despite substantial differences in their dynamic stabilities. These results suggest that the autoxidation reaction is more dependent on ligand accessibility than on the dynamic stability of the myoglobin.

Animals↗
PubMed1989

Piezoelectricity in cementum, dentine and bone.

Unlike the dental hard tissues, bone remodels when subjected to orthodontic forces. Bone is also piezoelectric (generates a surface electrical charge upon application of force). In dentine and cementum from sperm whale teeth (which gave samples of sufficient size), the existence and magnitude of piezoelectricity were examined and compared with human bone. Both dental tissues were found to be piezoelectric with coefficients of 0.027 and 0.028 pC/N, respectively; the coefficient of human bone was eight times greater (0.22 pC/N). Thus the strength of the piezoelectric effect was correlated with the known capacities of the tissues to undergo adaptive remodelling. This result is consistent with the theory that piezoelectricity mediates orthodontically induced alveolar remodelling.

Animals↗