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PubMed1981

Isolation and characterization of relaxin from the sand tiger shark (Odontaspis taurus).

A peptide with relaxin activity in guinea pigs but not in mice has been extracted from the ovaries of pregnant sand tiger sharks (Odontaspis taurus). The structural similarity of this peptide to porcine relaxin includes molecular size approximately 6000 daltons), number of chains, and possibly, disulfide cross-links. The relaxin-type peptide isolated from shark ovaries contains the amino acid residues tyrosine, proline, and histidine, which are absent in the porcine hormone. The amino acid composition of shark relaxin, therefore, resembles that of porcine insulin to a greater extent than does the amino acid composition of porcine relaxin. This finding supports the idea that shark relaxin may be a primitive relaxin that has undergone fewer mutations than porcine relaxin since the putative duplication of the insulin gene. The data presented here suggest that the putative duplication of the insulin gene, which might have given rise to relaxin, has occurred much earlier than the separation of sharks from the general branch of animals that eventually gave rise to mammals.

Amino Acids↗
PubMed1985

Calcitonin produces hypercalcemia in leopard sharks.

Calcitonin was detected by RIA in sera from four marine species, leopard sharks (Triakis semifasciata), horn sharks (Heterodontus francisci), thornback rays (Platyrhinoides triseriata), and kelp bass (Paralabrax clathratus). These animals have levels of calcitonin and calcium higher than freshwater and terrestrial species have. The administration of salmon calcitonin to bass (4 micrograms/kg BW) produced hypocalcemia and hypophosphatemia as has been reported for other bony vertebrates. In marked contrast, calcitonin produced a prompt hypercalcemia in sharks; the average was 9.8% increase in serum calcium in nine animals with no attendant change in phosphorus. These findings demonstrate that calcitonin can increase serum calcium in sharks. Because shark skeleton is composed of cartilage, this hypercalcemic effect of calcitonin does not require a bony skeleton.

Animals↗
PubMed2001

Shark cartilage as source of antiangiogenic compounds: from basic to clinical research.

The discovery that angiogenesis is a key condition for the growth of a tumor beyond a millimeter or two, brings about a new approach in the treatment of tumors using drugs able to inhibit the formation of new blood vessels. Also, it has been realized that antiangiogenic drugs can be useful in the treatment of other pathological processes, now classified as angiogenesis-dependent diseases. Initially, cartilage was considered as a possible natural source of antiangiogenic compounds due to its known avascular nature. To date, a number of in vitro and in vivo studies have suggested the existence of antiangiogenic and antitumor compounds in bovine and shark cartilage. However, the potential usefulness of shark cartilage in the treatment of cancer and other angiogenesis-dependent diseases have not been totally accepted due to (i) unsatisfactory patient outcome in clinical trials that have used shark cartilage in cancer patients, (ii) the lack of data that correlates bioavailability with pharmacological effects using oral shark cartilage. Thus, the objective of this review is to describe the main basic and clinical investigations reported in the literature, in which the antiangiogenic and/or antitumor properties of shark cartilage or of its extracts were evaluated. Possible explanations for conflicting results are discussed as well.

Angiogenesis Inhibitors↗
PubMed1995

Fusarium solani fungal infection of the lateral line canal system in captive scalloped hammerhead sharks (Sphyrna lewini) in Hawaii.

Two of five scalloped hammerhead sharks (Sphyrna lewini) captured May 1987 in Hawaii (USA) developed granulomatous exudative mycotic dermatitis localized in the lateral line canal system. The lesion initially was noted in the cephalic canals, but over a period of months extended into the lateral canal. Fusarium solani and Vibrio spp. were isolated from the canal exudate of both sharks. Bacterial colonies were not observed in the canal walls or surrounding tissues. Fusarium solani infection resulted in a chronic physical and behavioral deterioration of the two sharks; one shark was euthanized in September 1988 and the other in July 1989. This is the first report of Fusarium solani infection in the lateral line canal system and the third account in hammerhead sharks.

Animals↗
PubMed1999

Plasma steroid hormone profiles and reproductive biology of the epaulette shark, Hemiscyllium ocellatum.

Examination of the reproductive biology of the oviparous epaulette shark, Hemiscyllium ocellatum, was conducted on a wild population. Male sharks were found to reach maturity at between 55-60 cm total length (TL) and female sharks mature around 55 cm TL. Blood samples collected from mature male and female sharks were analyzed for sex steroid hormones to examine seasonal hormone patterns. Plasma samples were analyzed via radioimmunoassay techniques with female samples measured for estradiol, progesterone, and androgen concentrations, and male samples measured for androgen concentrations. Male androgen concentrations showed a single broad peak from July to October with maximum hormone concentrations (60 ng/ml) occurring in August. Male androgen concentrations were lowest in December-February (<20 ng/ml), and appeared to correlate with reproductive activity and water temperature. Female androgen concentrations were an order of magnitude lower than those for males and showed peaks in June (6 ng/ml) and December (8 ng/ml). Estradiol concentrations in females peaked during the months of September-November (0.5 ng/ml) coinciding with the egg laying period. Progesterone concentrations ranged up to 0.5 ng/ml prior to the mating season. Observations of ova size and egg production showed eggs develop in pairs and ova are ovulated at a size of 25-27 mm. Females lay eggs from August to January. Males were observed with swollen claspers from July through December, with the highest amount of sperm storage in the epididymis occurring between August through November. Our observations indicate that epaulette sharks in the waters near Heron Island mate from July through December. J. Exp. Zool. 284:586-594, 1999.

Androgens↗
PubMedDate not supplied

[Studies on the genotoxic effects of crude liver oils from 3 species of Mediterranean sharks by means of in vitro micronucleus test using human lymphocytes].

Lymphoid system tumours have been identified in two subjects who used to handle for several years mediterranean shark liver oil and squalen extracted from this oil. Moreover, scientific data, reported in 1959 by Kröning, show the induction of lymphoid tumours in C57 B1 mice after exposure of their skin to squalen. These observations rose the question of a possible mutagenic power of shark liver oil. In order to determine the genotoxicity of these oils, in vitro assays have been performed on crude hepatic oil of three species of mediterranean sharks: two benthic sharks, Centrophorus granulosus and Galeus melastomus, and one pelagic specie, Prionace glauca. Genotoxicity of oils have been assayed using a micronucleus test which can detected simultaneously clastogen and aneugen effects. The incubation of human cells with the hepatic crude oils of Centrophorus granulosus increases the rate of the binucleated micronucleated cell in a dose dependent manner. The mean micronucleated cell rate was 9.0%. +/- 1.1 in controls and increased up to 27,1%. +/- 4,0 for the highest concentrations of oil extracts. Similar results have been obtained with crude hepatic oils of Galeus melastomus and Prionace glauca. The results of this experimental study show that the crude liver oils of three species of sharks are genotoxic and confirm a high carcinogenic risk.

Adult↗
PubMed2000

[Dangerous sharks in tropical seas].

Sightseeing travel in tropical zones is a growing industry. The risks incurred by travelers depend on the destination, duration of stay, individual behavior, and type of leisure activity. Water sports expose visitors to encounters with dangerous marine animals. Shark attacks are rare but always serious occurrences. Divers should handle any shark, regardless of size, with due precaution. Prevention of shark attack depends on avoiding encounters by not attracting the attention of the shark and knowing the proper attitude to adopt in case an encounter should occur. Active and passive protection can be used, but each method has advantages and disadvantages depending on the situation. Rescue operations are difficult due to the gravity of injuries and their occurrence in a marine environment. This along with the nature of the aggressor explain that many attacks are immediately fatal. Wounds are often deep with involvement of bone, blood vessels, and nerves. A possible source of complication in survivors is infection, which can involve uncommon microorganisms associated with bacteria in sharks mouth or marine environment.

Animals↗
PubMed1981

Myoglobins of cartilaginous fishes III. Amino acid sequence of myoglobin of the shark Galeorhinus australis.

Myoglobin isolated from the red muscle of the school shark Galeorhinus australis was purified by gel filtration and ion-exchange chromatography. The amino acid sequence was determined following digestion with trypsin and purification of the peptides by paper ionophoresis and chromatography. Sequences of purified peptides were determined by the dansyl-Edman procedure and the peptides aligned by homology with the sequence of the myoglobin of the gummy shark Mustelus antarcticus. The two myoglobin sequences showed a marked similarity (16 differences), but both sequences showed approximately the same number of differences (68) from myoglobin of the Port Jackson shark Heterodontus portusjacksoni. There are 19 residues unique to three shark myoglobin sequences. As found with other fish myoglobins there are 148 residues with deletions of four residues at the amino terminal end as well as one residue in the CD region. The amino terminal residue is acetylated. The distal E7 histidine residue was found to be replaced by glutamine, as only previously reported for the myoglobin sequence of gummy shark.

Amino Acid Sequence↗
PubMed1994

[Fatal ichthyosarcotoxism after eating shark meat. Implications of two new marine toxins].

A fish poisoning involving 188 hospitalizations occurred in November 1993, in Manakara, a middle-sized town on the south-east coast of Madagascar, following the ingestion of shark. A single shark was involved in this poisoning and was identified as Carcharinus leucas. There was no unusual characteristic of this shark or its meat. The attack rate was about 100%. First clinical signs appeared within five to ten hours after ingestion. The patients presented neurological symptoms almost exclusively, the most prominent being a constant, severe ataxia. Gastrointestinal troubles, like diarrhoea and vomiting were rare. The overall case-fatality ratio was close to 30% among the 200 poisoned inhabitants. Search for similar poisoning previously reported in this area was negative, and fishermen in Manakara usually eat that kind of shark without mistrust. Bacteriological and chemical origins were eliminated. Two liposoluble toxins were isolated from the shark liver and tentatively named carchatoxin-A and -B respectively. They were distinct from ciguatoxin in chromatographic properties.

Adult↗
PubMed1993

The parasite assemblage in the spiral intestine of the shark Mustelus canis.

The parasite assemblage in the 8 chambers of the spiral intestine of 49 specimens of the shark Mustelus canis collected from Long Island Sound and off the coast of Virginia was investigated. Assemblages within host individuals were composed of up to 3 of 4 species of cestodes: the trypanorhynch species Prochristianella tumidula and Lacistorhynchus tenuis and the hooked tetraphyllidean species Calliobothrium verticillatum and Calliobothrium lintoni. Each individual shark hosted 1-3 (mean = 2.17) tapeworm species and 1-166 (mean = 34.3) tapeworm individuals. The assemblage consisted of 2 core species, 1 secondary species, and 1 satellite species. Analysis of variance (ANOVA) revealed significant differences among the areas of the 8 intestinal chambers. ANOVAs of worm density among chambers revealed that each of the 4 species exhibited site specificity within the spiral intestine. Least significant difference analysis revealed that each of the species, except C. verticillatum, was usually found in the anterior 3 chambers. Calliobothrium verticillatum was more commonly found in the middle region of the organ, centered around chamber 4. Horn's information index indicated that L. tenuis and C. lintoni had the greatest amount of overlap within the spiral intestine and the congeners C. lintoni and C. verticillatum had the least amount of overlap. No evidence of interaction among the species in this assemblage was found. In cases where observations could be made, neither cestode total length nor location within the spiral intestine appeared to be affected by the presence of other individuals or species respectively. There was some evidence of underutilized space; the posteriormost chamber was vacant in all 49 sharks examined and 91.8% of sharks had at least 2 vacant chambers. Linear regression revealed no relationship between shark total length and either total number of worms or total number of individuals of each of the 4 species of cestodes. The chi-square test revealed no evidence that the 4 species do not occur independently among host individuals. The number of species and number of individuals in M. canis were low when compared to the marine fish Sebastes nebulosus. These values for M. canis more closely resemble values for freshwater fishes. The comparatively much greater host specificity of these tapeworms may at least partially account for this difference. We propose that site specificity may have a phylogenetic component, in a manner similar to that of host specificity.

Analysis of Variance↗
PubMed1996

Forensic implications of the variation in morphology of marginal serrations on the teeth of the great white shark.

The teeth of the Great White Shark have been examined to ascertain whether there is any commonality in the arrangement or number of the marginal serrations (peaks) or, indeed, whether individual sharks have a unique pattern of shapes or size of the peaks. The teeth of the White Shark are characteristic in size and shape with serrations along almost the entire mesial and distal margins. This study has revealed no consistent pattern of size or arrangement of the marginal serrations that was sufficiently characteristic within an individual shark to serve as a reliable index of identification of a tooth as originating from that particular shark. Nonetheless, the serrations are sufficiently distinctive to enable the potential identification of an individual tooth as having been the cause of a particular bitemark.

Animals↗
PubMed1995

Isolation and characterization of the cDNA encoding the spiny dogfish shark (Squalus acanthias) form of cytochrome P450c17.

Cytochrome P450c17 is a key steroidogenic enzyme for the production of sex steroids in gonadal tissue and for cortisol production in adrenal tissue. This protein possesses two enzymatic activities. The 17 alpha-hydroxylase activity results in the introduction of a hydroxyl group at the 17 alpha-position. The resultant 17 alpha-hydroxylated, C21 pregnene is converted to a C19 androgen by the C17,20-lyase activity. A cDNA library was constructed from poly(A)-enriched mRNA isolated from spiny dogfish shark (Squalus acanthias) testis and ligated into EcoRI-cut lambda arms. The amplified library was screened using a bovine P450c17 cDNA probe and five positive clones were isolated. The described cDNA encompasses 23 bp of the 5'-untranslated region, a 1,527 bp open reading frame, and 414 bp of the 3'-untranslated region. A putative polyadenylation signal (AATAAA) is 18 bp from the poly(A) tail. Northern blot analysis showed a single transcript of 1.9 kb, thus indicating the isolated clone is a full-length cDNA. The deduced amino acid sequence of the shark form of P450c17 is 59% and 57% identical to the rainbow trout and chicken forms, respectively. The shark form is 43% to 46% identical to mammalian forms (rat, human, mouse, bovine, and porcine). There are large regions of extremely high identity shared among all the forms. The deduced shark 17 alpha-hydroxylase protein is 509 residues in length with a predicted weight of 57.2 kDa. Non-steroidogenic COS cells, transfected with the shark P450c17 cDNA, was capable of 17 alpha-hydroxylase and C17,20-lyase activities using both pregnenolone and progesterone as initial substrates.

Amino Acid Sequence↗
PubMed2002

Sharks: a potential source of antiangiogenic factors and tumor treatments.

Since angiogenesis is a key feature of tumor growth, inhibiting this process is one way to treat cancer. Cartilage is a natural source of material with strong antiangiogenic activity. This report reviews knowledge of the anticancer properties of shark cartilage and clinical information on drugs such as neovastat and squalamine. Because their entire endoskeleton is composed of cartilage, sharks are thought to be an ideal source of angiogenic and tumor growth inhibitors. Shark cartilage extract has shown antiangiogenic and antitumor activities in animals and humans. The oral administration of cartilage extract was efficacious in reducing angiogenesis. Purified antiangiogenic factors from shark cartilage, such as U-995 and neovastat (AE-941), also showed antiangiogenic and antitumor activity. AE-941 is under phase III clinical investigation. Squalamine, a low molecular weight aminosterol, showed strong antitumor activity when combined with chemotherapeutic materials. The angiogenic tissue inhibitor of metalloprotease 3 (TIMP-3) and tumor suppressor protein (snm23) genes from shark cartilage were cloned and characterized.

Journal Article↗
PubMed2006

Selection of cholera toxin specific IgNAR single-domain antibodies from a naïve shark library.

Shark immunoglobulin new antigen receptor (IgNAR, also referred to as NAR) variable domains (Vs) are single-domain antibody (sdAb) fragments containing only two hypervariable loop structures forming 3D topologies for a wide range of antigen recognition and binding. Their small size ( approximately 12kDa) and high solubility, thermostability and binding specificity make IgNARs an exceptional alternative source of engineered antibodies for sensor applications. Here, two new shark NAR V display libraries containing >10(7) unique clones from non-immunized (naïve) adult spiny dogfish (Squalus acanthias) and smooth dogfish (Mustelus canis) sharks were constructed. The most conserved consensus sequences derived from random clone sequence were compared with published nurse shark (Ginglymostoma cirratum) sequences. Cholera toxin (CT) was chosen for panning one of the naïve display libraries due to its severe pathogenicity and commercial availability. Three very similar CT binders were selected and purified soluble monomeric anti-CT sdAbs were characterized using Luminex(100) and traditional ELISA assays. These novel anti-CT sdAbs selected from our newly constructed shark NAR V sdAb library specifically bound to soluble antigen, without cross reacting with other irrelevant antigens. They also showed superior heat stability, exhibiting slow loss of activity over the course of one hour at high temperature (95 degrees C), while conventional antibodies lost all activity in the first 5-10min. The successful isolation of target specific sdAbs from one of our non-biased NAR libraries, demonstrate their ability to provide binders against an unacquainted antigen of interest.

Amino Acid Sequence↗
PubMed1999

Cloning, characterization, and functional expression of a CNP receptor regulating CFTR in the shark rectal gland.

In the shark, C-type natriuretic peptide (CNP) is the only cardiac natriuretic hormone identified and is a potent activator of Cl- secretion in the rectal gland, an epithelial organ of this species that contains cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels. We have cloned an ancestral CNP receptor (NPR-B) from the shark rectal gland that has an overall amino acid identity to the human homologue of 67%. The shark sequence maintains six extracellular Cys present in other NPR-B but lacks a glycosylation site and a Glu residue previously considered important for CNP binding. When shark NPR-B and human CFTR were coexpressed in Xenopus oocytes, CNP increased the cGMP content of oocytes (EC50 12 nM) and activated CFTR Cl- channels (EC50 8 nM). Oocyte cGMP increased 36-fold (from 0.11 +/- 0.03 to 4.03 +/- 0.45 pmol/oocyte) and Cl- current increased 37-fold (from -34 +/- 14 to -1,226 +/- 151 nA) in the presence of 50 nM CNP. These findings identify the specific natriuretic peptide receptor responsible for Cl- secretion in the shark rectal gland and provide the first evidence for activation of CFTR Cl- channels by a cloned NPR-B receptor.

Amino Acid Sequence↗
PubMed2006

Shark rectal gland vasoactive intestinal peptide receptor: cloning, functional expression, and regulation of CFTR chloride channels.

Vasoactive intestinal peptide (VIP) is a secretagogue that mediates chloride secretion in intestinal epithelia. We determined the relative potency of VIP and related peptides in the rectal gland of the elasmobranch dogfish shark and cloned and expressed the VIP receptor (sVIP-R) from this species. In the perfused rectal gland, VIP (5 nM) stimulated chloride secretion from 250 +/- 66 to 2,604 +/- 286 microeq x h(-1) x g(-1); the relative potency of peptide agonists was VIP > PHI = GHRH > PACAP > secretin, where PHI is peptide histidine isoleucine amide, GHRH is growth hormone-releasing hormone, and PACAP is pituitary adenylate cylase activating peptide. The cloned sVIP-R from shark rectal gland (SRG) is only 61% identical to the human VIP-R1. It maintains a long, extracellular NH2 terminus with seven cysteine residues, and has three N-glycosylation sites and eight other residues implicated in VIP binding. Two amino acids considered important for peptide binding in mammals are not present in the shark orthologue. When sVIP-R and the CFTR chloride channel were coexpressed in Xenopus oocytes, VIP increased chloride conductance from 11.3 +/- 2 to 127 +/- 34 microS. The agonist affinity for activating chloride conductance by the cloned receptor was VIP > GHRH = PHI > PACAP > secretin, a profile mirroring that in the perfused gland. The receptor differs from previously cloned VIP-Rs in having a low affinity for PACAP. Expression of both sVIP-R and CFTR mRNA was detected by quantitative PCR in shark rectal gland, intestine, and brain. These studies characterize a unique G protein-coupled receptor from the shark rectal gland that is the oldest cloned VIP-R.

Amino Acid Sequence↗
PubMed1991

Identification of C-type natriuretic peptide in heart of spiny dogfish shark (Squalus acanthias).

An unidentified atrial natriuretic peptide (ANP)-like substance is the principal hormone regulating NaCl secretion in the shark rectal gland, an epithelial model tissue for hormone-sensitive secondary active chloride transport. Antibodies to mammalian ANP do not recognize the prohormone of marine species. The polymerase chain reaction (PCR) was used to isolate a partial cDNA encoding the shark heart natriuretic peptide. Using this partial sequence as a probe, the full-length clone [882 base pairs (bp)] was obtained from a shark heart cDNA library. Amino acids 119-135 are similar to the recently identified peptide sequences of porcine C-type natriuretic peptide (CNP) and killifish brain natriuretic peptide isolated from the brain of these species. Mature shark heart CNP terminates at the second cysteine residue and lacks a COOH-terminal extension, in contrast to cardiac ANP-like peptides of all other species. The primary amino acid sequence of the shark heart prepro-CNP is distinctly different from all other cardiac natriuretic peptides. Amplification of genomic DNA spanning the coding region produced a 1.5-kb product, indicating the presence of at least one intron. Sequencing confirmed the presence of two exons of 90 and 315 bp, separated by a 1.1-kb intron. This is the first report of a cDNA encoding in nonneuronal tissue. Elasmobranch CNP may represent a primordial form of ANP-like peptides that evolved as an adaptation to environmental osmoregulatory stress.

Amino Acid Sequence↗
PubMed1998

Effects of prey size and mobility on prey-capture kinematics in leopard sharks triakis semifasciata

Recent work on teleosts suggests that attack behaviors or kinematics may be modified by a predator on the basis of the size of the prey or the ability of the prey to sense predators and escape capture (elusivity). Sharks are generally presumed to be highly visual predators; thus, it is reasonable to expect that they might also be capable of such behavioral modulation. In this study, I investigated the effect of prey item size and type on prey-capture behavior in leopard sharks (Triakis semifasciata) that had been acclimated to feeding in the laboratory. Using high-speed video, sharks were filmed feeding on two sizes of the same prey item (thawed shrimp pieces) and two potentially more elusive prey items (live earthworms and live mud shrimp). In leopard sharks, little effect of prey elusivity was found for kinematic variables during prey capture. However, the large proportion of successful captures of the live prey suggests that they did not prove to be truly elusive prey items for the leopard shark. There were significant size effects on prey-capture kinematics, with the larger non-elusive items inducing greater head expansion during prey capture. Ram-suction index values also indicated that strikes on large, non-elusive prey had a significantly larger suction component than strikes on similar small prey items. This finding is interesting given that the two sizes of non-elusive prey item offered no differential challenge in terms of a performance consequence (reduced capture success).

Journal Article↗