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PubMed1997

A 1H NMR comparative study of the structure of the critical packing interfaces between helix and non-helical region in various ligation states of sperm whale myoglobin.

NMR signals arising from HisB5 N(delta)H and HisEF5 N(epsilon)H protons in sperm whale met-aquo, met-azido, met-imidazole and deoxy myoglobins have been assigned. HisB5 N(delta)H proton and N(epsilon) atom are hydrogen-bonded to peptide carbonyl oxygen of AspB1 and HisGH1 N(epsilon)H proton, respectively. This unique triad hydrogen bond stabilizes the interface between the B helix and the GH comer, which is critical to the stability of its protein folding. HisEF5 N(epsilon)H proton is hydrogen-bonded to one of the side-chain carboxyl oxygen atoms of AspH18 and this internal hydrogen bond stabilizes the orientation of the EF corner, the hinge region between the heme binding E and F helices, relative to the H helix. The assigned His NH proton signals were used as probes to investigate possible structural differences of these critical interfaces among various ligation states of myoglobin. Shift differences of the signals among the proteins strongly suggested that the AspB1-HisB5-HisGH1 and HisEF5-AspH18 hydrogen bonds are affected by heme ligation. The present study demonstrates the presence of a structural correlation between the active site and the critical interfaces in myoglobin.

Animals↗

PubMed1998

A comparison of human and killer whale platelet fatty acid composition.

Activation of blood platelets and their subsequent aggregation results from the interactions of several complex metabolic pathways. Considered to be of critical importance are the platelet lipids. Subsequent to platelet activation, several membrane lipids undergo hydrolysis and the free fatty acids are metabolized to prostanoids which mediate platelet function in response to vascular injury. It is conceivable then, that differences in platelet membrane fatty acid content could result in significant differences in platelet responses to aggregatory stimuli, especially between species. The objective of this study was to identify specific differences in fatty acid content between human and killer whale platelets. Blood was collected, washed platelets were prepared, and platelet fatty acids were extracted. Methyl esters of the extracted fatty acids were analyzed by gas chromatography and reported as relative concentrations. Analysis of the data revealed significant differences between the two species for several relevant fatty acids, i.e. 16:0 (P < 0.05), and 18:0, 18:1, 18:2, and 20:4 (P < 0.001). The differences in platelet fatty acid composition and concentration may explain at least some of the differences in platelet function which have previously been identified between these species.

Adult↗

PubMed1999

The anatomy of the killer whale middle ear (Orcinus orca).

The paper first reviews our present understanding of the functional morphology of the odontocete (toothed whale) ear. The tympano-periotic complex forming the ear region consists of a ventral bowl-shaped tympanic bone in direct contact with the surrounding soft tissues and the incident sound, and a dorsal periotic bone containing the inner ear. Apparently sound brings the tympanic bone, and especially its thin tympanic plate, into vibration. The ossicles in the air-filled middle ear cavity form a bridge from the tympanic plate to the periotic bone connecting the vibrating plate to the oval window and the inner ear. Our computer tomography (CT) sections and camera lucida drawings reveal two hitherto unknown features of the odontocete ear, both of them of potential relevance to sound reception and impedance matching. (1) It is well known that, in addition to the ossicular chain, two other bone structures connect the tympanic to the periotic bone. We show that the most delicate parts of these extra-ossicular connections consist of thin and folded bony sheets which apparently allow compliance in the tympano-periotic bone contacts and enable plate vibration in relation to the periotic bone. (2) The round head of the malleus, in combination with a fitting round depression on the periotic side, seems to form a joint. We propose that this (hypothetical) joint, together with the adjacent structures, forms a lever producing an amplification of the vibration velocity at the level of the oval window.

Animals↗

PubMed1967

Periodate oxidation of sperm-whale myoglobin and the role of the methionine residues in the antigen-antibody reaction.

1. Oxidation of sperm-whale metmyoglobin and its apoprotein with periodate has been investigated under various conditions of pH and temperature to find those under which the reagent acted with specificity. 2. At pH6.8 and 22 degrees consumption of periodate ceased in 3(1/2)hr. at 43 moles of periodate/mole of myoglobin. The two methionine residues, the two tryptophan residues, the three tyrosine residues and two histidine residues were oxidized; serine increased in the hydrolysates from 6 to 9 residues/mol. 3. At pH5.0 and 22 degrees , consumption levelled off in 4(1/2)hr. at 26 moles of periodate/mole of myoglobin and resulted in the modification of the two methionine residues, the two tryptophan residues, the three tyrosine residues and two histidine residues; serine increased from 6 to 7 residues/mol. and, also, ferrihaem suffered considerable oxidation. 4. Oxidation at pH5.0 and 0 degrees resulted at completion (4hr.) in the consumption of 22 moles of periodate/mole of myoglobin and in the modification of the methionine, tyrosine and tryptophan residues. Spectral studies indicated oxidation of the haem group. This derivative reacted very poorly with rabbit antisera to MbX (the major component no. 10 obtained by CM-cellulose chromatography; Atassi, 1964). 5. Oxidation of apomyoglobin at pH5.0 and 0 degrees was complete in 4hr. with the consumption of 7.23 moles of periodate/mole of apoprotein. The rate of oxidation in decreasing order was: methionine; tryptophan; tyrosine; and after 7hr. of reaction the following residues/mol. were oxidized: methionine, 2.0; tryptophan, 1.6; tyrosine, 0.99. No peptide bonds were cleaved. Metmyoglobin prepared from the 7hr.-oxidized apoprotein showed that the reactivity with antisera to MbX had diminished considerably. 6. Milder oxidation of apoprotein (2 molar excess of periodate, pH5.0, 0 degrees , 2hr.) resulted in the modification of 1.66 residues of methionine/mol. Metmyoglobin prepared from this apoprotein was identical with native MbX spectrally, electrophoretically and immunochemically. It was concluded that the methionine residues at positions 55 and 131 were not essential parts of the antigenic sites of metmyoglobin.

Animals↗

PubMed1968

Analysis of the visible spectra of some sperm-whale ferrimyoglobin derivatives.

The visible spectra of sperm-whale metmyoglobin and its fluoride, hydroxide, cyanide, azide, cyanate, formate, acetate, thiocyanate and nitrite derivatives were measured and resolved into Gaussian components. A linear correlation between the intensity of the band at 18500cm.(-1) and the magnetic susceptibility is found, suggesting that the band is the beta-band of the low-spin form. The analysed intensity of this band can be used to determine the relative proportions of the two spin states present. The spectra of the pure high-spin and low-spin components of each derivative can be reconstructed. Correlations between the energy of the near-infrared band and the visible bands of the high-spin form, and between the position and intensity of the 16000cm.(-1) band, are demonstrated. The relationship between the position of the Soret band and the magnetic susceptibility can be explained in terms of the equilibrium between spin states.

Acetates↗

PubMed2001

Lingual ultrastructure of the long-finned pilot whale (Globicephala melas).

Microscopic studies on the cetacean tongue are limited and, to date, only a few ultrastructural reports on dolphins have been published. This report presents the initial description of the lingual ultrastructure of the long-finned pilot whale. The lingual integumental surface was smooth, lacking papillae, although flaking of outer stratum corneum cells could be observed at high resolution. The keratinocytes of the stratum spinosum of the epidermis resembled those of cetacean skin on other regions of the body. The similarities included the presence of cytoplasmic lipid droplets around the nuclei of stratutm spinosum cells, a lingual feature not seen in terrestrial mammals. Keratin intermediate filaments were numerous and occasionally formed aggregates of circular whorls. At cell surfaces, bundles of keratin intermediate filaments were frequently observed inserting into desmosomal plaques. Pigment granules were not evident and organelles were sparse. Stratum corneal cells contained nuclear remnants (parakeratosis) and small multivesicular bodies, and the corneal layer was approximately 18 cells thick. The nuclei of the stratum basale keratinocytes possessed exceptionally numerous and deep clefts. The dermis was nondistinctive. The skeletal muscle of the tongue was arranged in widely separated fasiculi containing small numbers of muscle fibres. Typical fine structure of skeletal muscle bands and tubular elements was observed by transmission electron microscopy.

Animals↗

PubMed1982

The causes of whale strandings.

Strandings of cetaceans (whales and dolphins) is probably the result of a sequence of events rather than a simple cause-and-effect relationship. Strandings of solitary animals are more common than massed and generally thought to be due to severe disease. Mass strandings (three to several hundred individuals) are less common, but very spectacular. Mass strandings are considered to be due to a complex of factors in which behavioural factors are probably important, severe disease making an unimportant, or negligible contribution. There may be interaction of physical conditions (weather, tides, currents, coastline) and biological conditions (predators, feeding, disturbance of echolocation and disease) which result in stranding. One postulated reason for stranding is that it is dependent on instinctive behaviour, deeply located in subcortical areas of the brain, which provokes cetaceans to seek refuge ashore when under stress.

Journal Article↗

PubMed1984

Treatment of a strandling whale (Kogia breviceps).

A stranded young pygmy sperm whale was nursed for four days in a pool, fed an artificial diet via a stomach tube and injected intramuscularly with antibiotics and corticosteroids. Initially unable to maintain her balance or to swim, she showed progressive improvement and by the third day could swim and dive quite strongly. Early on the fifth day her condition rapidly worsened and she died. An autopsy revealed extensive deep bruising in the head and mid-abdominal regions consistent with injuries sustained during the stranding.

Journal Article↗

PubMed2003

Surface properties of the skin of the pilot whale Globicephala melas.

On the skin surface of delphinids small biofoulers are challenged to high shear water flow and liquid-vapor interfaces of air-bubbles during jumping. This state of self-cleaning is supported by the even, nano-rough gel-coated epidermal surface of the skin. The present study focussed on the intercellular evolution of gel formation and the chemical composition of the gel smoothing the skin surface of the pilot whale, Globicephala melas, using X-ray photoelectron spectroscopy (XPS) in combination with cryo-scanning electron microscopy (CSM), and transmission electron microscopy (TEM). In the superficial layer of the epidermis, the stratum corneum, intercellular material was shown by electron optical methods to assemble from smaller into larger covalently cross-linked aggregates during the transit of the corneocytes towards the skin surface. XPS measurements showed that the surface of the skin and the intercellular gel included approximately the same amounts of polar groups (especially, free amines and amides) and non-polar groups, corresponding to the presence of lipid droplets dispersed within the jelly material. It was concluded from the results that the gel-coat of the skin surface is a chemically heterogeneous skin product. The advantages of chemically heterogeneous patches contributing to the ablation of traces of the biofouling process are discussed.

Animals↗

PubMed1965

The crystal-solution problem of sperm whale myoglobin.

The central question to be discussed in this paper is whether the structure established for sperm whale myoglobin in the crystalline state is the same as that of the protein in solution. As judged by its ultraviolet optical rotatory dispersion, the helical content of metmyoglobin in solution does not differ from that in the crystal, 77 per cent. Although an uncertainty of about +/-5 per cent must attach to this result, it excludes many alternative arrangements of the polypeptide chain. The folding of the chain may be further restricted to the basic form seen in the crystal if the dimensions of the molecule in solution and the interactions of specific chemical groups are taken into account. Since the rotatory dispersion of metmyoglobin is constant with respect to ionic strength, and since the dispersions of reduced and oxymyoglobin reveal no change in helical content upon their formation from metmyoglobin, one may infer that the structure of the protein is largely maintained both as it dissolves and during its reversible combination with oxygen. The crystallographic model of myoglobin thus offers a structural basis for attempting to explain its physiological function in solution. The relevance of this conclusion to the crystal-solution problems presented by other species of protein is then best seen in the light of common factors that govern the equilibrium of all proteins between crystal and solution.

Animals↗

PubMed2004

Reproductive physiology and development of artificial insemination technology in killer whales (Orcinus orca).

Research was conducted to define the basic reproductive physiology of killer whales (Orcinus orca) and to use this knowledge to facilitate the development of artificial insemination procedures. The specific objectives were 1) to determine the excretory dynamics of urinary LH and ovarian steroid metabolites during the estrous cycle; 2) to evaluate the effect of an exogenously administered, synthetic progesterone analog on reproductive hormone excretion; 3) to validate the use of transabdominal ultrasound for ovarian evaluation and timing of ovulation; 4) to examine the quality of semen after liquid storage and cryopreservation; and 5) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of 41 follicular phases and 26 complete cycles from five females, estrous cycles were 41 days long and comprised a 17-day follicular phase and a 21-day luteal phase. A consistent temporal relationship was observed between peak estrogen conjugates and the LH surge, the latter of which occurred approximately 0.5 days later. Two animals placed on oral altrenogest (three separate occasions for 30, 17, and 31 days, respectively) excreted peak urinary estrogen concentrations 25 days after withdrawal that were followed by sustained elevations in urinary pregnanediol-3alpha-glucuronide excretion. Mean preovulatory follicle diameter was 3.9 cm (n = 6), and ovulation occurred 38 h (n = 5) after the peak of the LH surge. Based on visual estimates of motility, liquid-stored semen maintained 92% of its raw ejaculate sperm motility index (total progressive motility x kinetic rating [0-5 scale, where 0 = no movement and 5 = rapid progressive movement]) when held at 4 degrees C for 3 days postcollection. Semen cryopreserved using a medium freezing rate demonstrated good postthaw total motility (50%), progressive motility (94%), and kinetic rating (3.5). Insemination during eight estrous cycles resulted in three pregnancies (38%), two from liquid-stored and one from cryopreserved semen. Two calves were delivered after gestation lengths of 552 and 554 days, respectively. These data demonstrate the potential of noninvasive endocrine monitoring combined with serial ultrasonography to improve our understanding of the reproductive biology of cetaceans. This fundamental knowledge was essential for ensuring the first successful conceptions, resulting in live offspring, using artificial insemination in any cetacean species.

Acrosome↗

PubMed2002

Low worldwide genetic diversity in the killer whale (Orcinus orca): implications for demographic history.

A low level of genetic variation in mammalian populations where the census population size is relatively large has been attributed to various factors, such as a naturally small effective population size, historical bottlenecks and social behaviour. The killer whale (Orcinus orca) is an abundant, highly social species with reduced genetic variation. We find no consistent geographical pattern of global diversity and no mtDNA variation within some regional populations. The regional lack of variation is likely to be due to the strict matrilineal expansion of local populations. The worldwide pattern and paucity of diversity may indicate a historical bottleneck as an additional factor.

Alleles↗

PubMed2004

The interaction of outgoing echolocation pulses and echoes in the false killer whale's auditory system: evoked-potential study.

Brain auditory evoked potentials (AEP) associated with echolocation were recorded in a false killer whale Pseudorca crassidens trained to accept suction-cup EEG electrodes and to detect targets by echolocation. AEP collection was triggered by echolocation pulses transmitted by the animal. The target was a hollow aluminum cylinder of strength of -22 dB at a distance from 1 to 8 m. Each AEP record was obtained by averaging more than 1000 individual records. All the records contained two AEP sets: the first one of a constant latency and a second one with a delay proportional to the distance. The timing of these two AEP sets was interpreted as responses to the transmitted echolocation pulse and echo, respectively. The echo-related AEP, although slightly smaller, was comparable to the outgoing click-related AEP in amplitude, even though at a target distance as far as 8 m the echo intensity was as low as -64 dB relative to the transmitted pulse in front of the head. The amplitude of the echo-related AEP was almost independent of distance, even though variation of target distance from 1 to 8 m influenced the echo intensity by as much as 36 dB.

Acoustic Stimulation↗

PubMed2005

Invariance of evoked-potential echo-responses to target strength and distance in an echolocating false killer whale.

Brain auditory evoked potentials (AEPs) were recorded in a false killer whale Pseudorca crassidens trained to accept suction-cup EEG electrodes and to detect targets by echolocation. AEP collection was triggered by echolocation pulses transmitted by the animal. The target strength varied from -22 to -40 dB; the distance varied from 1.5 to 6 m. All the records contained two AEP sets: the first one of a constant latency (transmission-related AEP) and a second one with a delay proportional to the distance (echo-related AEP). The amplitude of echo-related AEPs was almost independent of both target strength and distance, though combined variation of these two parameters resulted in echo intensity variation within a range of 42 dB. The amplitude of transmission-related AEPs was independent of distance but dependent on target strength: the less the target strength, the higher the amplitude. Recording of transmitted pulses has not shown their intensity dependence on target strength. It is supposed that the constancy of echo-related AEP results from variation of hearing sensitivity depending on the target strength and release of echo-related responses from masking by transmitted pulses depending on the distance.

Animals↗

PubMed2005

Types, distribution, and seasonal occurrence of sounds attributed to Bryde's whales (Balaenoptera edeni) recorded in the eastern tropical Pacific, 1999-2001.

Vocalizations resembling known Bryde's whale sounds were recorded on autonomous hydrophones at seven sites in the eastern tropical Pacific. Five short (<3 s) low-frequency (<80 Hz) "phrase" types were observed. "Swept alternating tonal" phrases included a 37-Hz tone and often a 25-16-Hz downswept tone, while "nonswept alternating tonal" phrases had a predominant tone at 29 Hz and often additional tones at 16 and 47 Hz. Alternating tonal phrases were found in 79% of the total hours in which phrases were detected, and occurred primarily at the eastern hydrophone sites. "Burst-tonal" phrases included tones that were often preceded by a wideband burst of noise. The "low burst-tonal" phrase contained tones at 19 and 30 Hz, and was detected at five of the hydrophone sites. The "high burst-tonal" phrase included a 42-Hz tone and was observed only on the northwestern hydrophones. A single "harmonic tone" phrase type was observed that included a fundamental tone at 26 Hz and at least two harmonics; this phrase was observed exclusively at the eastern hydrophone stations. This opportunistic survey has shown that acoustics is an effective means of studying this poorly understood, pelagic balaenopterid.

Acoustics↗

PubMed2005

Behavioral and auditory evoked potential audiograms of a false killer whale (Pseudorca crassidens).

Behavioral and auditory evoked potential (AEP) audiograms of a false killer whale were measured using the same subject and experimental conditions. The objective was to compare and assess the correspondence of auditory thresholds collected by behavioral and electrophysiological techniques. Behavioral audiograms used 3-s pure-tone stimuli from 4 to 45 kHz, and were conducted with a go/no-go modified staircase procedure. AEP audiograms used 20-ms sinusoidally amplitude-modulated tone bursts from 4 to 45 kHz, and the electrophysiological responses were received through gold disc electrodes in rubber suction cups. The behavioral data were reliable and repeatable, with the region of best sensitivity between 16 and 24 kHz and peak sensitivity at 20 kHz. The AEP audiograms produced thresholds that were also consistent over time, with range of best sensitivity from 16 to 22.5 kHz and peak sensitivity at 22.5 kHz. Behavioral thresholds were always lower than AEP thresholds. However, AEP audiograms were completed in a shorter amount of time with minimum participation from the animal. These data indicated that behavioral and AEP techniques can be used successfully and interchangeably to measure cetacean hearing sensitivity.

Acoustic Stimulation↗

PubMed1999

Quantifying complex patterns of bioacoustic variation: use of a neural network to compare killer whale (Orcinus orca) dialects.

A quantitative measure of acoustic similarity is crucial to any study comparing vocalizations of different species, social groups, or individuals. The goal of this study was to develop a method of extracting frequency contours from recordings of pulsed vocalizations and to test a nonlinear index of acoustic similarity based on the error of an artificial neural network at classifying them. Since the performance of neural networks depends on the amount of consistent variation in the training data, this technique can be used to assess such variation from samples of acoustic signals. The frequency contour extraction and the neural network index were tested on samples of one call type shared by nine social groups of killer whales. For comparison, call similarity was judged by three human subjects in pairwise classification tasks. The results showed a significant correlation between the neural network index and the similarity ratings by the subjects. Both measures of acoustic similarity were significantly correlated with the groups' association patterns, indicating that both methods of quantifying acoustic similarity are biologically meaningful. An index based on neural network analysis therefore represents an objective and repeatable means of measuring acoustic similarity, and allows comparison of results across studies, species and time.

Acoustics↗

PubMed1969

"Blow" of the pilot whale.

A captive pilot whale emptied as much as 88 percent of lung gas passively, without the aid of expiratory muscles. Level or decreasing pressures in the esophagus during expiration, and in the blowhole at the onset of expiration, revealed the driving force of expiration to be solely elastic recoil. Active muscular reexpansion of the lungs ensued immediately. Expiration and inspiration were completed in about I second.

Animals↗