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Effects of dietary marine oils and olive oil on fatty acid composition, platelet membrane fluidity, platelet responses, and serum lipids in healthy humans.

The influence of various dietary marine oils and olive oil on fatty acid composition of serum and platelets and effects on platelets and serum lipids were investigated as part of an extensive study of the effects of these oils on parameters associated with cardiovascular/thrombotic diseases. Healthy volunteers (266) consumed 15 mL/d of cod liver oil (CLO); whale blubber oil (refined or unrefined); mixtures of seal blubber oil and CLO; or olive oil/CLO for 12 wk. In the CLO, seal oil/CLO, and whale oil groups, serum levels of eicosapentaenoic acid (EPA) were increased. In platelets, EPA was increased in the CLO, seal/CLO, and olive oil/CLO groups. The localization of n-3 polyunsaturated fatty acids in the triacylglycerols did not seem to influence their absorption. Intake of oleic acid is poorly reflected in serum and platelets. No significant differences in triacylglycerols (TG), total cholesterol, or high density lipoprotein cholesterol were observed, even though TG were reduced in the CLO, CLO/seal oil, and whale oil groups. Mean platelet volume increased significantly in both whale oil groups and the CLO/olive oil group. Platelet count was significantly reduced in the refined whale oil group only. Lipopolysaccharide-stimulated blood tended to generate less thromboxane B2 in CLO, CLO/seal, and CLO/olive groups. The whale oils tended to reduce in vivo release of beta-thromboglobulin. In conclusion, intake of various marine oils causes changes in platelet membranes that are favorably antithrombotic. The combination of CLO and olive oil may produce better effects than these oils given separately. The changes in platelet function are directly associated with alterations of fatty acid composition in platelet membranes.

Adolescent↗

1H NMR investigation of the heme cavity of elephant (E7 Gln) met-cyano-myoglobin. Evidence for a B-helix phenylalanine interaction with bound ligand.

A combination of one- and two-dimensional NMR experiments has been used to identify and spatially locate the heme pocket residues in the paramagnetic, low spin, met-cyano complex of elephant myoglobin. In addition to assigning resonances of the conserved residues, we have also assigned Gln64 (E7) and an aromatic ring designated PheA whose side chain is inserted into the heme pocket, as found earlier for elephant carbonmonoxy-myoglobin and oxy-myoglobin (Yu, L. P., La Mar, G. N., and Mizukami, H. (1990) Biochemistry 29, 2578-2585). The assigned conserved proximal side residues (Leu89(F4), Ala90(F5), His93(F8), His97(FG3), Ile99(FG5), Leu104(G5), Phe138(H15), and Tyr146(H23)) and conserved distal side residues (Phe43(CD1), Thr67(E10), Val68(E11), and Ala71(E14)) in elephant met-cyano-myoglobin are found to have orientations similar to those in sperm whale met-cyano-myoglobin. The observed dipolar connectivities and dipolar shift pattern for the substituted Gln64(E7) place the Gln in the heme pocket oriented toward the iron, as found for His64(E7). The conserved structural elements demand that the inserted PheA originate from the B-helix (i.e. Phe27 or Phe33). Dipolar contacts between the inserted PheA and the conserved residues Phe43(CD1), Val68(E11), Ile107(G8), and Gln64(E7), place PheA in the position occupied by the B10 residue in sperm whale myoglobin (Mb), with the larger size of the PheA side chain as compared to the replaced Leu being accommodated by the vacancy that occurs in sperm whale Mb. The paramagnetic induced relaxation places PheA in van der Waals contact with the bound ligand. Hence we conclude that the B10 position of elephant Mb is occupied by a Phe, and this substitution relative to sperm whale Mb is responsible for the low autoxidation rate and low reduction potential of elephant Mb. A reduced autoxidation rate has been reported for a sperm whale synthetic point mutant Leu29(B10) --> Phe (Carver, T. E., Brantley, R. E., Jr., Singleton, E. W., Arduini, R. M., Quillin, M. L., Phillips, G. N., and Olson, J. S. (1992) J. Biol. Chem. 267, 14443-14450). The published sequence of elephant Mb places B-helix Phe residues at position 27(B8) and 33(B14), but a Phe at neither of these positions can account for the observed NMR properties. Since a large proportion of the substitutions in elephant relative to sperm whale Mb, and some of the least conservative, occur in the B-helix, neither a structurally perturbed B-helix nor an error in the sequence can be discounted.

Animals↗

Effect of the proportion of organic material in bone on thermal decomposition of bone mineral: an investigation of a variety of bones from different species using thermogravimetric analysis coupled to mass spectrometry, high-temperature X-ray diffraction, and Fourier transform infrared spectroscopy.

Thermogravimetric analysis linked to mass spectrometry (TGA-MS) shows changes in mass and identifies gases evolved when a material is heated. Heating to 600 degrees C enabled samples of bone to be classified as having a high (cod clythrum, deer antler, and whale periotic fin bone) or a low (porpoise ear bone, whale tympanic bulla, and whale ear bone) proportion of organic material. At higher temperatures, the mineral phase of the bone decomposed. High temperature X-ray diffraction (HTXRD) showed that the main solids produced by decomposition of mineral (in air or argon at 800 degrees C to 1000 degrees C) were beta-tricalcium phosphate (TCP) and hydroxyapatite (HAP), in deer antler, and CaO and HAP, in whale tympanic bulla. In carbon dioxide, the decomposition was retarded, indicating that the changes observed in air and argon were a result of the loss of carbonate ions from the mineral. Fourier transform infrared (FTIR) spectroscopy of bones heated to different temperatures, showed that loss of carbon dioxide (as a result of decomposition of carbonate ions) was accompanied by the appearance of hydroxide ions. These results can be explained if the structure of bone mineral is represented by [Formula: see text] where V(Ca) and V(OH) correspond to vacancies on the calcium and hydroxide sites, respectively, and 2-x-y = 0.4. This general formula is consistent in describing both mature bone mineral (i.e., whale bone), with a high Ca/P molar ratio, lower HPO4(2-) content, and higher CO3(2-) content, and immature bone mineral (i.e., deer antler), with a low Ca/P ratio, higher HPO4(2-), and lower CO3(2-) content.

Animals↗

Human exposure to contaminants in the traditional Greenland diet.

The traditional diet is a significant source of contaminants to people in Greenland, although contaminant levels vary widely among species and tissue from very low in many to very high in a few. Our study has included cadmium, mercury, selenium, polychlorinated biphenyls (PCB), dichlorophenyltrichloroethane (DDT), chlordane, hexachlorocyclohexanes (HCH), chlorobenzenes, dieldrin and toxaphene in the major species and tissues consumed by Greenlanders. In general, the levels of these are very low in terrestrial species and in muscle of many marine species. High organochlorines concentrations are typically found in blubber of marine mammals and high metal levels in liver and kidney of seals and whales. In this study, the mean intakes of cadmium, chlordanes and toxaphene significantly exceed 'acceptable/tolerable intakes' (ADI/TDI) by a factor between 2.5 and 6. Mean intakes of mercury, PCB and dieldrin also exceed ADI/TDI by up to approximately 50%. However as these figures are mean intakes and as variation in both food intake and contaminant levels is large, the variation of contaminant intake among individuals is also large, and some individuals will be exposed to significantly higher intakes. The mean intakes of DDT, HCH and chlorobenzenes are well below the ADI/TDI values, and it seems unlikely that the TDI for these contaminants normally is exceeded in the Greenland population. The evaluation of contaminant intake in this study points to seal muscle, seal liver, seal kidney, seal blubber and whale blubber as the dominant contributors of contaminants in the traditional diet. Levels in liver from Greenland halibut, snow crab, king eider, kittiwake, beluga and narwhal and kidney of beluga and narwhal are also high but were, with the exception of toxaphene in Greenland halibut liver, not important sources in this study, because they were eaten in low quantities. A way to minimize contaminant intake would be to avoid or limit the consumption of diet items with high contaminant levels. If we assume a traditional diet composition in this study without fish liver, bird liver, seal liver, seal kidney, seal blubber, whale liver, whale kidney and whale blubber, the intake of all contaminants would be below the TDIs for these. This will result in a reduction of the intake of the amount of traditional food of only 24-25%, and it is not likely that this changed diet will result in deficiency of minerals, vitamins or other nutritional compounds.

Adolescent↗

Time-lapsed microstructural imaging of bone failure behavior.

Many bones within the axial and appendicular skeleton are subjected to repetitive loading during the course of ordinary daily activities. If this loading is of sufficient magnitude or duration, failure of the bone tissue may result. Until recently the structural analysis of these fractures has been limited to two-dimensional sections. Due to the inherent destructiveness of this method, dynamic assessment of fracture progression has not been possible. An image-guided technique to analyze structural failure has been developed utilizing step-wise micro-compression in combination with time-lapsed micro-computed tomographic imaging. This technique allows, for the first time, direct three-dimensional visualization and quantification of fracture initiation and progression on the microscopic level and relates the global failure properties of trabecular bone to those of the individual trabeculae. The goals of this project were first to design and fabricate a novel micro-mechanical testing system, composed of a micro-compression device and a material testing and data acquisition system; and second, to validate the testing system to perform step-wise testing of trabecular bone specimens based on image-guided failure analysis. Due to the rate dependant properties of bone, stress relaxation was a concerning factor with respect to the step-wise testing method. In order to address these concerns, the results of the step-wise testing method were compared to those obtained from a conventional continuous test (considered to be the gold standard for the step-wise compressive mechanical testing) over the same total strain range and testing conditions. This was performed using porous aluminum alloy samples with highly reproducible and homogenous structural properties as well as trabecular bone samples from a single whale vertebra. Five cylinders from aluminum foam and trabecular whale bone each were compressed and imaged in a step-wise fashion from 0% to 20% strain at intervals of 2%, 4%, 8%, 12%, 16% and 20%. Mechanical properties obtained from the continuous and step-wise methods were not significantly different for both aluminum foam and whale bone specimens (p>0.05). Both testing methods yielded very similar stress-strain graphs with almost identical elastic and plastic regions with overlaying standard error bars for both whale bone and aluminum foam specimens. This was further concurred by performing regression analyses between the stress data from both testing methods (r(2)=0.98 for whale bone and aluminum foam specimens). Animations of fracture initiation and progression revealed that failure always occurred in local bands with the remaining regions of the structure largely unaffected independent of structure type. In conclusion, we found step-wise micro-compression to be a valid approach for image-guided failure assessment (IGFA) with high precision and accuracy as compared to classical continuous testing. We expect findings from upcoming studies of IGFA of human vertebral bone to improve our understanding of the relative importance of densitometric, morphological, and loading factors in the etiology of spontaneous fractures of the spine. Eventually, this improved understanding may lead to more successful approaches to the prevention of age-related fatigue fractures.

Animals↗

Marine mammals as global pollution indicators for organochlorines.

Blubber tissues of the following marine mammals differing in their geographic distribution (North Atlantic, North Pacific, Bering Sea/Arctic Ocean), trophic level, and feeding habits were analyzed for their organochlorine contents: two seal species (harbor seals--Phoca vitulina, northern fur seals--Callorhinus ursinus), three toothed whale species (belukha whales--Delphinapterus leucas, one common dolphin--Delphinus delphis, one pilot whale--Globicephala melaena), and one baleen whale species (a bowhead whale--Balaena mysticetus). As xenobiotics were quantified the seven indicator congeners of the polychlorinated biphenyls (PCB 28, 52, 101, 118, 138, 153, and 180), three isomers of the hexachlorocyclohexanes (alpha-, beta- and gamma-HCH) as well as six components of the DDT-group (4,4'-DDT, 4,4'-DDD, 4,4'-DDE, 2,4'-DDT, 2,4'-DDD, and 2,4'-DDE). When comparing the xenobiotic levels of these marine mammals, it showed that the animals from the western North Atlantic were contaminated about 15 times more with organochlorines than the animals from the eastern North Pacific and the Bering Sea/Arctic Ocean. The total organochlorine burden, the 4,4'-DDE-percentage as well as the metabolic PCB patterns correlate with the trophic levels of the marine mammals studied. The quantitative analyses were done by high-resolution capillary gas chromatography with electron capture detection (HRGC/ECD) whereas the analyses of the metabolic PCB patterns were done by high-resolution capillary gas chromatography and mass selective detection (HRGC/MSD).

Adipose Tissue↗

Assignment of 1H NMR resonances of histidine and other aromatic residues in met-, cyano-, oxy-, and (carbon monoxy)myoglobins.

The resolved 1H NMR resonances of the aromatic region in the 270-MHz NMR spectrum of sperm whale, horse, and pig metmyoglobin (metMb) have been assigned, including the observable H-2 and H-4 histidine resonances, the tryptophan H-2 resonances, and upfield-shifted resonances from one tyrosine residue. The use of different Mb species, carboxymethylation, and matching of pK values allows the assignment of the H-4 resonances, which agree in only three cases out of seven with scalar-correlated two-dimensional NMR spectroscopy assignments by others. The conversion to hydroxymyoglobin at high pH involves rearrangements throughout the molecule and is observed by many assigned residues. In sperm whale ferric cyanomyoglobin, nine H-2 and eight H-4 histidine resonances have been assigned, including the His-97 H-2 resonance and tyrosine resonances from residues 103 and 146. The hyperfine-shifted resonances from heme and near-heme protons observe a shift with a pK = 5.3 +/- 0.3 (probably due to deprotonation of His-97, pK = 5.6) and another shift at pK = 10.8 +/- 0.3. The spectrum of high-spin ferrous sperm whale deoxymyoglobin is very similar to that of metMb, which allows the assignment of seven surface histidine H-2 and H-4 resonances and also resonances from the two tryptophan residues and one tyrosine. In diamagnetic sperm whale (carbon monoxy)myoglobin (COMb), 10 His H-2 and 11 His H-4 resonances are observed, and 8 H-2 and 9 H-4 resonances are assigned, including His-64 H-4, the distal histidine. This important resonance is not observed in sperm whale oxymyoglobin, which in general shows very similar titration curves to COMb. Histidine-36 shows unusual titration behavior in the paramagnetic derivatives but normal behavior in the diamagnetic derivatives, which is discussed in the accompanying paper [Bradbury, J. H., & Carver, J. A. (1984) Biochemistry (following paper in this issue)].

Animals↗

Imidazole is a sensitive probe of steric hindrance in the distal pockets of oxygen-binding heme proteins.

The FixL heme-based sensor, despite its low affinity for oxygen, is much more reactive than myoglobin toward the large polar ligand imidazole. To determine which features of a myoglobin heme pocket favor binding of imidazole, we have measured binding of this ligand to the FixL heme domain, elephant myoglobin, wild-type sperm whale myoglobin, and sperm whale myoglobins having alanine, valine, threonine, glutamine, leucine, phenylalanine, or tryptophan substitutions of the distal (E7) histidine residue. Except for histidine, the association rate constants dropped more than 3000-fold as the volume of the E7 side chain, at position 64, was expanded from alanine (10(6) M-1 s-1) to phenylalanine (10(3) M-1 s-1). There was inhibition of imidazole binding due to displacement of coordinated water from H64 and H64Q sperm whale myoglobins, where the E7 side chain hydrogen bonds directly to the bound ligand. The imidazole dissociation rate constants varied less dramatically and less consistently with any single factor, though they were measurably decreased by hydrogen bonding to an E7 glutamine or histidine. On the whole, the results for the sperm whale myoglobin E7 substitutions show that the rate constants for imidazole binding are useful and sensitive indicators of steric hindrance and polar interactions in the distal pockets of myoglobins. The combined effects of the glutamine 64 and phenylalanine 29 in elephant myoglobin largely account for its increased imidazole association and dissociation rate constants, respectively, compared to those of sperm whale myoglobin. An unhindered distal pocket not competent to stabilize positive poles is indicated by the large imidazole association (>/=10(4) M-1 s-1) and dissociation (>/=50 s-1) rate constants, parameters that are characteristic of FixL.

Amino Acid Substitution↗

Myoglobin structure and regulation of solvent accessibility of heme pocket.

The effects of heme removal on the molecular structure of tuna and sperm whale myoglobin have been investigated by comparing the solvent accessibility to the heme pocket of the two proteins with that of the corresponding apoproteins. Although the heme microenvironment of tuna myoglobin is more polar than that of sperm whale myoglobin, the accessibility of solvent to heme is identical in the two proteins as revealed by thermal perturbation of Soret absorption. The removal of heme produces loss of helical folding and increase of solvent accessibility but the effects are rather different for the two proteins. More precisely, the loss of helical structure upon heme removal is 50% for tuna myoglobin and 15% for sperm whale myoglobin; moreover, the solvent accessibility of the heme pocket of tuna apomyoglobin is 2-3-fold greater than that of sperm whale apomyoglobin. These results have been explained in terms of the lack of helical folding in segment D, the structural organization of which may have a relevant effect in regulating the accessibility of ligands to the heme. The effects produced by charged quenchers reveal that the ligand path from the surface of the molecule to the ion atom of the heme involves a positively charged residue which may reasonably be identified as Arg-45 (sperm whale myoglobin) or Lys-41 (tuna myoglobin) on the basis of recent X-ray crystallographic information.

Animals↗

Determination of relative binding affinity of influenza virus N9 sialidases with the Fab fragment of monoclonal antibody NC41 using biosensor technology.

The relative binding affinities of influenza virus N9 sialidase from term and whale with the Fab fragment of monoclonal antibody NC41 were determined using biosensor technology (Pharmacia BIAcoreTM). The apparent association and dissociation rate constants were measured in real time for the interaction of the Fab with both sialidases, the Fab being immobilised on the sensor surface. Although three-dimensional structural studies have shown that there are no apparent structural differences between the term and whale N9 sialidase epitopes to which the NC41 Fab binds, the apparent binding constant for the interaction with tern N9 sialidase was approximately 2.4-fold higher than that with whale N9 sialidase. The kinetic analysis showed that the association rate constant for the binding of whale N9 sialidase was higher than that for tern N9 sialidase (12.0 x 10(4) M-1 s-1 compared to 4.3 x 10(4) M-1 s-1) and the dissociation rate constants for the whale N9-sialidase-Fab complex were approximately 6-fold higher than for the tern N9-sialidase-Fab complex. Furthermore, kinetic analysis of the dissociation reaction showed that it was composed of two stages, an initial, faster rate followed by a late, slower rate. The values of the relative affinity constants calculated using the initial dissociation rate constant were similar to the values measured at equilibrium in the BIAcore and those determined in true solution equilibrium studies using sedimentation equilibrium. The late, slower, dissociation rate constant yielded affinity constants significantly higher than those obtained by true solution methods.

Animals↗

DNA registers of legally obtained wildlife and derived products as means to identify illegal takes.

The exploitation and sale of wildlife species that are endangered in only part of their range present regulators with the critical challenge of separating legal from illegal takes. Wildlife DNA registers created from tissue samples of legally obtained individual wildlife specimens can address this problem by allowing managers to identify unregistered (presumably illegally obtained) specimens. We tested the effectiveness of the only current, fully operational wildlife DNA register of individual genetic profiles collected from legally caught minke whales (Balaenoptera acutorostrata). Twenty minke whale tissue samples collected at markets in Norway and 2 additional samples collected from beached minke whales in Denmark were genotyped at 12 loci used by the Norwegian minke whale DNA register Genetic profiles of these samples then were compared against the 2676 individual profiles deposited in the Norwegian register The high number of genetic markers used to identify individuals in our study allowed consistent matching of sample and reference profiles despite an overall error rate (due to experimental and interlaboratory data standardization) estimated at 0.015 per locus. Of the 22 test samples only the 2 Danish samples failed to match an existing profile in the Norwegian minke whale DNA register Our results show that the basic principle of wildlife DNA registers can work in a real-life situation. The strength of wildlife DNA registers lies in their ability to unambiguously identify unregistered specimens with the aid of sensitive genetic methods that enable analysis of highly processed or degraded tissue samples. Our study also highlights a number of methodological problems such as laboratory errors and interlaboratory data standardization, which need be addressed to ensure a successful implementation of wildlife DNA registers.

Animal Identification Systems↗

Evidence of Brucella infection in marine mammals in the North Atlantic Ocean.

Between 1983 and 1996 a total of 1386 samples of serum were taken from four species of seal and three species of whale in the waters west of Iceland, the area of pack-ice north-west of Jan Mayen, the northern coast of Norway and the Kola Peninsula, the waters west of Svalbard, and the Barents Sea; they were tested for the presence of anti-Brucella antibodies with an indirect ELISA (protein G conjugate). The positive sera were re-tested with classical brucellosis serological tests, such as the serum agglutination test, the EDTA-modified serum agglutination test, the Rose Bengal test, and the complement fixation test, as well as an anti-complement ELISA. Anti-Brucella antibodies were detected in all the species investigated, except for the bearded seal (Erignathus barbatus), with the following prevalences: hooded seals (Cystophora cristata) 35 per cent; harp seals (Phoca groenlandica) 2 per cent; ringed seals (Phoca hispida) 10 per cent; minke whales (Balaenoptera acutorostrata) 8 per cent; fin whales (Balaenoptera physalus) 11 per cent; and sei whales (Balaenoptera borealis) 14 per cent. An isolate belonging to the genus Brucella was obtained from the liver and spleen of one of the seropositive minke whales. The findings suggest that antibodies against the surface lipopolysaccharide of Brucella species are widely distributed among marine mammals in the North Atlantic Ocean.

Agglutination Tests↗

Mercury, food webs, and marine mammals: implications of diet and climate change for human health.

We modeled the flow of methyl mercury, a toxic global pollutant, in the Faroe Islands marine ecosystem and compared average human methyl mercury exposure from consumption of pilot whale meat and fish (cod, Gadus morhua) with current tolerable weekly intake (TWI) levels. Under present conditions and climate change scenarios, methyl mercury increased in the ecosystem, translating into increased human exposure over time. However, we saw greater changes as a result of changing fishing mortalities. A large portion of the general human population exceed the TWI levels set by the World Health Organization [WHO; 1.6 microg/kg body weight (bw)], and they all exceed the reference dose (RfD) of 0.1 microg/kg bw/day set by the U.S. Environmental Protection Agency (EPA; equivalent to a TWI of 0.7 microg/kg bw). As a result of an independent study documenting that Faroese children exposed prenatally to methyl mercury had reduced cognitive abilities, pregnant women have decreased their intake of whale meat and were below the TWI levels set by the WHO and the U.S. EPA. Cod had approximately 95% lower methyl mercury concentrations than did pilot whale. Thus, the high and harmful levels of methyl mercury in the diet of Faroe Islanders are driven by whale meat consumption, and the increasing impact of climate change is likely to exacerbate this situation. Significantly, base inflow rates of mercury into the environment would need to be reduced by approximately 50% to ensure levels of intake below the WHO TWI levels, given current levels of whale consumption.

Adolescent↗

Arterial Windkessels in marine mammals.

In marine mammals, the aortic arch is enlarged relative to the descending aorta to varying degrees in different species. The ratio of maximal diameter of the arch to that of the thoracic aorta is about 2.3 in the harbour seal (Phoca vitulina), 3.6 in the Weddell seal (Leptonychotes weddelli) and 3.2 in the fin whale (Balaenoptera physalus), compared with only 1.4 in the dog. This anatomical specialisation probably provides increased volume capacitance in the arterial circulation as an adaptation to diving bradycardia. Data on the morphometric and mechanical properties of aortic tissues from seals and fin whale are compared. In the harbour seal, more than 80% of the volume change in the entire thoracic aorta that results from a pressure pulse occurs in the bulbous arch, and this is more than 90% in the Weddell seal and fin whale. The enhanced capacitance of the arch in the harbour seal is primarily due to its larger diameter, as the relative wall thickness and elasticity of the arch and thoracic aorta are the same. A similar situation appears to exist in the larger Weddell seal, although extrapolation of the pressure-volume curves suggests that the arch might be somewhat less stiff than the thoracic aorta. In addition to being greatly expanded, the aortic arch of the fin whale is also much more distensible than the relatively thin-walled and much stiffer descending aorta. At the estimated mean blood pressure, the elastic modulus of this vessel is 12 MPa, or 30 times that of the aortic arch. The major haemodynamic consequence of this type of arterial modification is that the aortic arch acts as a Windkessel, i.e. the capacitance of the aorta is increased significantly close to the heart, leading to a reduced characteristic impedance and probably reduced pulsatility in the descending aorta. In the extreme case of the whale, the arterial capacitance is shifted entirely to the arch, and the impedance change at the entrance to the thoracic aorta is so high that this probably represents the major reflection site in the arterial tree.

Adaptation, Physiological↗

Epidemiological analysis of persistent organochlorine contaminants in cetaceans.

Information is provided to test the hypothesis that organochlorines introduced into the environment since the early 1940s could threaten the reproductive potential of baleen whales and other cetaceans. Comparisons are made using data on the role of organochlorines in a model system, the Great Lakes region of North America, and in model animals, including humans, pinnipeds, and other wildlife. DDT and PCB are used as model organochlorines with the caveat that there may be thousands of other chemicals in the environment also involved. Improved sensitivity in analytical quantification of synthetic chemicals in biological tissue has been accompanied by an increase in knowledge about biochemical processes that control development and function. The effects described in this review are the result of disrupted gene expression, not damage to the gene. The mechanisms of action of the organochlorines reveal their ability to affect developing organisms at very low concentrations during critical life stages: embryonic, fetal, and early postnatal. Exposure during early development can disrupt the organization of the endocrine, reproductive, immune and nervous systems, effecting irreversible damage that may not be expressed until the individuals reach adulthood. The recent discovery that human sperm count is declining worldwide at a rate of 1 x 10(6) sperm/(mL.yr) suggests common exposure to estrogen-like chemicals during prenatal and early postnatal development. This raises concern for other top predator species that also share the same exposure. Periods of intense feeding followed by long periods of fasting are common among species of baleen whales. This unique strategy places the embryonic and nursing calves in vulnerable positions, because under both situations maternal blood levels are elevated as a result of absorption from food intake or as a result of mobilization as fat is metabolized. Estimates of Toxic Equivalents (TEQs) based on the occurrence of four PCB congeners (118, 183, 153, 180) in sigma PCB reported in whales are highest for St. Lawrence belugas and Faroe Island long-finned pilot whales. This conservative approach reveals that some whale species are within the range of enzyme-induced TEQs at which effects have been associated with adverse health effects in other aquatic species. The epidemiological approach was used for analysis because it was developed to handle multiple exposure scenarios in which direct causal links are virtually impossible to isolate. The analysis includes the tenets of timeorder, strength of association, specificity of cause and effect, consistency, coherence, and predictive performance.

Animals↗

Mitogenomic analyses provide new insights into cetacean origin and evolution.

The evolution of the order Cetacea (whales, dolphins, porpoises) has, for a long time, attracted the attention of evolutionary biologists. Here we examine cetacean phylogenetic relationships on the basis of analyses of complete mitochondrial genomes that represent all extant cetacean families. The results suggest that the ancestors of recent cetaceans had an explosive evolutionary radiation 30-35 million years before present. During this period, extant cetaceans divided into the two primary groups, Mysticeti (baleen whales) and Odontoceti (toothed whales). Soon after this basal split, the Odontoceti diverged into the four extant lineages, sperm whales, beaked whales, Indian river dolphins and delphinoids (iniid river dolphins, narwhals/belugas, porpoises and true dolphins). The current data set has allowed test of two recent morphological hypotheses on cetacean origin. One of these hypotheses posits that Artiodactyla and Cetacea originated from the extinct group Mesonychia, and the other that Mesonychia/Cetacea constitutes a sister group to Artiodactyla. The current results are inconsistent with both these hypotheses. The findings suggest that the claimed morphological similarities between Mesonychia and Cetacea are the result of evolutionary convergence rather than common ancestry.

Animals↗

Morphology, molecules, and the phylogenetics of cetaceans.

Recent phylogenetic analyses of cetacean relationships based on DNA sequence data have challenged the traditional view that baleen whales (Mysticeti) and toothed whales (Odontoceti) are each monophyletic, arguing instead that baleen whales are the sister group of the odontocete family Physeteridae (sperm whales). We reexamined this issue in light of a morphological data set composed of 207 characters and molecular data sets of published 12S, 16S, and cytochrome b mitochondrial DNA sequences. We reach four primary conclusions: (1) Our morphological data set strongly supports the traditional view of odontocete monophyly; (2) the unrooted molecular and morphological trees are very similar, and most of the conflict results from alternative rooting positions; (3) the rooting position of the molecular tree is sensitive to choice of artiodactyls outgroup taxa and the treatment of two small but ambiguously aligned regions of the 12S and 16S sequences, whereas the morphological root is strongly supported; and (4) combined analyses of the morphological and molecular data provide a well-supported phylogenetic estimate consistent with that based on the morphological data alone (and the traditional view of toothed-whale monophyly) but with increased bootstrap support at nearly every node of the tree.

Animals↗

The correct molecular weight of myoglobin, a common calibrant for mass spectrometry.

Myoglobins from horse heart muscle, horse skeletal muscle and sperm whale are widely used as calibration standards or test compounds for various mass spectrometric methodologies. In all such cases reported in the literature, a molecular weight value is used (16,950.5 and 17,199, respectively) which is based on the assumption that amino acid 122 in this 153 amino-acid-long protein is asparagine, overlooking a published suggestion that it is aspartic acid instead. Since the mass assignment accuracy for matrix-assisted laser desorption mass spectrometry is reported to be +/- 0.01% and for electrospray ionization +/- 0.0025%, and error of one mass unit in approximately 17,000 would be significant. The mass-to-charge ratio of ions of the tryptic peptide encompassing amino acid 122 derived from commercially available horse heart and horse skeletal myoglobins, the apomyoglobin of the latter, and the tryptic and chymotryptic peptide of sperm whale myoglobin proved that in both proteins amino acid 122 is indeed aspartic acid, rather than asparagine. This finding was further confirmed by the collision-induced dissociation spectra of the [M + H]+ ions of the tryptic peptides from the horse myoglobins and the chymotriptic peptide from sperm whale myoglobin. Thus, the correct molecular weight of horse myoglobin is 16,951.49 and that of the sperm whale protein is 17,199.91.

Amino Acid Sequence↗