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Description of a new species of the genus Raphidascaroides Yamaguti, 1941 (Nematoda: Heterocheilidae (from the shark, Sphyrna ztgaena (Linnaeus) at Visakhapatnam, India.

A new species of the nematode genus Raphidascaroides Yamaguti, 1941 obtained from the intestine of hammer-headed shark, Sphyrna zygaena (Linnaeus) from Visakhapatnam is described. The specimens do not agree with the description of known species of the genus. Raphidascaroides zygaenai has been established by body size, length of oesophagus and oesophageal appendix, size of spicules, number and arrangement of caudal papillae, vulva position and structure of tail end.

Animals↗

A new species of Phoreiobothrium (Cestoidea:Tetraphyllidea) from the great hammerhead shark Sphyrna mokarran and its implications for the evolution of the onchobothriid scolex.

Phoreiobothrium manirei n. sp. is described from the spiral intestines of 3 immature individuals of the great hammerhead shark Sphyrna mokarran collected off of the west coast of Florida. This cestode is unusual in that it possesses 4 muscular papillae on the anterior margin of the accessory sucker. Numerous small protrusions with central cilium-like projections were conspicuous throughout the posterior margin of each bothridium. This species possesses the unique combination of bothridia posteriorly subdivided into subloculi and hooks with extended bases and 2 rather than 3 prongs. In fact, the hooks are strikingly like those of species in the genera Dicranobothrium and Platybothrium. This combination of characters suggests that the bifid hook condition is plesiomorphic with respect to the trifid hook condition within the Onchobothriidae. These data further suggest that the triloculated bothridial condition is plesiomorphic relative to the biloculated condition.

Animals↗

Histopathological reactions of the blue shark, Prionace glauca, to postlarvae of Hepatoxylon trichiuri (Cestoda: Trypanorhyncha: Hepatoxylidae) in relationship to scolex morphology.

Postlarvae of the cestode Hepatoxylon trichiuri (Holten, 1802) were found attached to the surface of viscera of Prionace glauca (Linnaeus, 1758) taken from Atlantic coastal waters off the south coast of Massachusetts, USA. Gross anatomy of the attachment site shows a quadripartite rim surrounding a deep pit with holes corresponding to the penetration site of each tentacle. Hyperplasia of the liver capsule into outgrowths at the attachment site conform to the attachment of the bothridia. The cuboidal epithelium of the liver capsule became columnar forming papillary outgrowths, with a dense fibrotic reaction beneath the attachment site and infiltration by leukocytes and pigment containing granulocytic cells. Blood sinusoids beneath the attachment site are greatly enlarged. Postlarvae attached to the surface of the epigonal organs of three blue sharks were not accompanied by reactions. SEM examination of the scolex of H. trichiuri postlarvae revealed fused pairs of bothridia within infolded muscular lateral rims, porose tegument devoid of microtriches and a bilateral plane of symmetry in the tentacular armature.

Animals↗

Effect of U-995, a potent shark cartilage-derived angiogenesis inhibitor, on anti-angiogenesis and anti-tumor activities.

BACKGROUND: A potent angiogenesis inhibitor, U-995, has been purified from the cartilage of the blue shark (Prionace glauca). U-995 is composed of two single peptides with molecular mass of 10 and 14 kDa, respectively. MATERIALS AND METHODS: U-995 was designed to study human umbilical vein endothelial cell (HUVEC) migration and proliferation in vitro and angiogenesis induced by TNF alpha in chicken chorioallantoic membrane (CAM). Furthermore, we determined the ability of U-995 to inhibiting tumor cell growth and metastasis. RESULTS: U-995 (15 and 30 micrograms/ml) markedly inhibited HUVEC migration and, at 15-50 micrograms/ml produced a dose-dependent decline in [3H]-thymidine incorporation. 30 and 50 micrograms/ml of U-995, when added to TNF alpha-induced angiogenesis caused discontinuous and disrupted blood vessels. Moreover, U-995 (30 micrograms/ml) markedly prevented collagenase-induced collagenolysis. In addition, when 200 micrograms U-995 was injected i.p. into mice it suppressed sarcoma-180 cell growth and B16-F10 mouse melanoma cell metastasis in vivo. CONCLUSIONS: These results suggest that the anti-angiogenic effects of U-995 may be be due to interference with the proliferation and migration of HUVECs as well as inhibition of collagenolysis, thereby leading to inhibition of both angiogenesis and tumor cell growth.

Allantois↗

Brainstem neurons with descending projections to the spinal cord of two elasmobranch fishes: thornback guitarfish, Platyrhinoidis triseriata, and horn shark, Heterodontus francisci.

We studied two cartilaginous fishes and described their brainstem supraspinal projections because most nuclei in the reticular formation can be identified that way. A retrogradely transported tracer, horseradish peroxidase or Fluoro-Gold, was injected into the spinal cord of Platyrhinoidis triseriata (thornback guitarfish) or Heterodontus fransisci (horn shark). We described labeled reticular cells by their position, morpohology, somatic orientation, dendritic processes, and laterality of spinal projections. Nineteen reticular nuclei have spinal projections: reticularis (r.) dorsalis, r. ventralis pars alpha and beta, r. gigantocellularis, r. magnocellularis, r. parvocellularis, r. paragigantocellularis lateralis and dorsalis, r. pontis caudalis pars alpha and beta, r. pontis oralis pars medialis and lateralis, r. subcuneiformis, r. peduncularis pars compacta, r. subcoeruleus pars alpha, raphe obscurus, raphe pallidus, raphe magnus, and locus coeruleus. Twenty nonreticular nuclei have spinal projections: descending trigeminal, retroambiguus, solitarius, posterior octaval, descending octaval, magnocellular octaval, ruber, Edinger-Westphal, nucleus of the medial longitudinal fasciculus, interstitial nucleus of Cajal, latral mesencephalic complex, periventricularis pretectalis pars dorsalis, central pretectal, ventromedial thalamic, posterior central thalamic, posterior dorsal thalamic, the posterior tuberculum, and nuclei B, F, and J. The large number of distinct reticular nuclei with spinal projections corroborates the hypothesis that the reticular formation of elasmobranches is complexly organized into many of the same nuclei that are found in frogs, reptiles, birds, and mammals.

Animals↗

Short-term effects of 3,5,3'-triiodothyronine on the intermediary metabolism of the dogfish shark Squalus acanthias: evidence from enzyme activities.

Plasma 3,5,3'-triiodothyronine (T3) concentration decreased significantly (P < 0.05), during 1-5 days of captivity, from levels in the freshly caught dogfish shark Squalus acanthias. The short-term effects of T3 treatment on the intermediary metabolism of S. acanthias were measured in the gill, kidney, liver, and white muscle. Animals were kept for 1-5 days before experimentation. Three hours after an intraperitoneal injection with either a low T3 dose (8.3 pmol T3/kg fish) or a high T3 dose (830 pmol T3/kg fish), selected enzymes of amino acid metabolism, lipid catabolism, ketone body metabolism, glycolysis, and oxidative metabolism were measured. Activity of enzymes of amino acid metabolism and lipid catabolism increased significantly (P < 0.05) in the liver of fish treated with a low T3 dose. The low dose of T3 apparently influences glycolysis as pyruvate kinase activity significantly increase (P < 0.05) in the kidney and white muscle.

Animals↗

Cadmium disrupts the signal transduction pathway of both inhibitory and stimulatory receptors regulating chloride secretion in the shark rectal gland.

The heavy metal cadmium causes nephrotoxicity and alters the transport function of epithelial cells. In the shark rectal gland, chloride secretion is regulated by secretagogues and inhibitors acting through receptors coupled to G proteins and the cyclic AMP-protein kinase A pathway. We examined the effects of cadmium on the response to the inhibitory peptide somatostatin (SRIF), and to the stimulatory secretagogues forskolin and vasoactive intestinal peptide (VIP). In control experiments, SRIF (100 nM) entirely inhibited the chloride secretory response to 10 microM forskolin (maximum chloride secretion with forskolin 1984 +/- 176 microEq/h/g; with forskolin + SRIF 466 +/- 93 microEq/h/g, P < 0.001). Cadmium (25 microM) entirely reversed the inhibitory response to SRIF (chloride secretion 2143 +/- 222 microEq/h/g) and caused an overshoot (2917 +/- 293 microEq/h/g) that exceeded the response to forskolin (P < 0.01). Cadmium also enhanced forskolin-stimulated chloride secretion (2628 +/- 418 vs. 1673 +/- 340 microEq/h/g, P < 0.02) and reversed the declining phase of the forskolin response. Cadmium had a concentration-dependent, biphasic effect on the response to VIP. Cd (10-100 microM) increased both chloride secretion and tissue cyclic AMP content, whereas higher concentrations (1 mM) inhibited chloride secretion and cyclic AMP accumulation. Our findings provide evidence that Cd disrupts the signal transduction pathways of both inhibitory receptors and secretagogues regulating cAMP mediated transport in an intact epithelia. The results are consistent with direct effects of cadmium on adenylate cyclase and/or phosphodiesterase activity in this marine epithelial model.

Adenylyl Cyclases↗

Anatomy of the feeding apparatus of the nurse shark, Ginglymostoma cirratum.

The anatomy of the feeding apparatus of the nurse shark, Ginglymostoma cirratum, was investigated by gross dissection and computer axial tomography. The labial cartilages, jaws, jaw suspension, muscles, and ligaments of the head are described. Palatoquadrate cartilages articulate with the chondrocranium caudally by short, laterally projecting hyomandibulae and rostrally by ethmoorbital articulations. Short orbital processes of the palatoquadrates are joined to the ethmoid region of the chondrocranium by short, thin ethmopalatine ligaments. In addition, various ligaments, muscles, and the integument contribute to the suspension of the jaws. When the mouth is closed and the palatoquadrate retracted, the palatine process of the palatoquadrate is braced against the ventral surface of the nasal capsule and the ascending process of the palatoquadrate is in contact with the rostrodorsal end of the suborbital shelf. When the mandible is depressed and the palatoquadrate protrudes slightly rostroventrally, the palatoquadrate moves away from the chondrocranium. A dual articulation of the quadratomandibular joint restricts lateral movement between the mandible and the palatoquadrate. The vertically oriented preorbitalis muscle spans the gape and is hypothesized to contribute to the generation of powerful crushing forces for its hard prey. The attachment of the preorbitalis to the prominent labial cartilages is also hypothesized to assist in the retraction of the labial cartilages during jaw closure. Separate levator palatoquadrati and spiracularis muscles, which are longitudinally oriented and attach the chondrocranium to the palatoquadrate, are hypothesized to assist in the retraction of the palatoquadrate during the recovery phase of feeding kinematics. Morphological specializations for suction feeding that contribute to large subambient suction pressures include hypertrophied coracohyoideus and coracobranchiales muscles to depress the hyoid and branchial arches, a small oral aperture with well-developed labial cartilages that occlude the gape laterally, and small teeth. Copyright 1999 Wiley-Liss, Inc.

Journal Article↗

Trimethylamine oxide and the maintenance of volume of dogfish shark rectal gland cells.

Determinants of the steady-state vol of the dogfish shark (Squalus acanthias) rectal gland cells were studied. The cellular levels of trimethylamine oxide (TMAO) in fresh tissue and slices incubated aerobically 60 min in standard (TMAO-free) elasmobranch saline were close to those in the plasma (71 +/- 5 mM S.E.M.); therefore, under these conditions, the cell membrane appears to be impermeable to this solute. However, depolarization of the cells in high-K+ media produced a rapid loss of TMAO. Thus, TMAO is a major, effectively impermeant solute in the rectal gland cells. The osmolarity of cell solutes in tissue water (fresh and incubated slices) did not differ significantly from values in the plasma or incubation medium, demonstrating the absence of an osmotic pressure gradient across the cell membrane. An analysis of a simple model of cell solutes under steady-state conditions shows that the presence of an (effectively) impermeant osmolyte decreases the cellular concentration of bulk cations. The analysis is consistent with available observations on the distribution of cell Na+ and K+ in tissues containing high concentrations of (nitrogeneous) osmolytes. One simplifying assumption of the model, i.e., identity (or closeness) of the respective reflection coefficients sigma for Na+ and K+ passage through the cell membranes could not be verified. Compared to available data on the steady-state cellular fluxes of 42K+ in slices of the rectal gland, the uptake of 22Na+ by the tissue was slow (the derived rate constant k' = 0.017 min-1, i.e., about one tenth of that for K+).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Shark testis model: stage-dependent functions and the regulation of spermatogenesis.

In mammals, a single Sertoli cell nurtures 3-4 successive generations of germ cells. Thus, it is not possible to study this cell type at a single spermatogenic stage. In the dogfish shark Squalus acanthias, a single cohort of Sertoli cells remains associated with a germ cell clone throughout its development. Moreover, different germ cell stages are topographically segregated within the testis and can be easily staged by transilluminationmicroscopy. Recently, we have developed methods for the isolation and culture of spermatocysts (Sertoli/germ cell units) and Sertoli cells only from pre-meiotic, meiotic, and post-meiotic stages of germ cell development. Here, we present data that illustrate the feasibility of using the Squalus testis model for characterizing stage-related biochemical changes in Sertoli cells.

Animals↗

Hepatoprotective effects of the shark bile salt 5beta-scymnol on acetaminophen-induced liver damage in mice.

The hepatoprotective effect of the shark bile salt 5beta-scymnol has been studied in the model of acute hepatotoxicity induced by administration of acetaminophen (APAP, paracetamol). 5beta-Scymnol at doses of 20, 35, and 70 mg/kg intraperitoneally (ip) decreased significantly the serum activity of alanine aminotransferase, sorbitol dehydrogenase, and lactate dehydrogenase (p < 0.05) caused by APAP treatment (350 mg/kg ip) alone. The highest dose of 5beta-scymnol remained hepatoprotective when administered 4 hr after the APAP overdose. N-Acetylcysteine (NAC) is protective against APAP-induced hepatotoxicity at 250 and 500 mg/kg (ip) when administered up to 3 hr after APAP overdose, as shown by a significant reduction in serum enzyme activity. Coadministration of 5beta-scymnol (70 mg/kg) and NAC (250 mg/kg) also reduced serum enzyme levels and histopathological effects; however, a similar level of hepatoprotection was conferred by 5beta-scymnol treatment alone. In addition, 5beta-scymnol has potent hydroxyl radical quenching activity as it markedly inhibited deoxyribose degradation in a ferrous/ascorbate Fenton reaction system. These results indicate a possible role for the use of 5beta-scymnol, either alone or concomitant with NAC, in the prevention of hepatic necrosis following toxic doses of APAP.

Acetaminophen↗

Evidence for the presence of A1 and A2 adenosine receptors in the ventral aorta of the dogfish shark, Squalus acanthias.

Isolated, endothelium-free rings of vascular smooth muscle (VSM) from the ventral aorta of the dogfish shark, Squalus acanthias, were used to examine the vasoactive effects of various adenosine agonists. Cumulative addition of 2-chloroadenosine (2 Cl-ADO) over the concentration range 10 nM-1 mM resulted in a biphasic response, with a significant increase in tension at 1 microM and a more significant decline in tension at 100 microM and 1 mM, suggesting that this tissue may possess both A1 and A2 adenosine receptors. N6-Cyclopentyladenosine (N-6 CPA) and N6-(2-phenylisopropyl)adenosine, R(-)isomer (R-PIA), generally considered to be more A1 specific, also produced slight, but significant increases in tension, but only at relatively high concentrations. The more specific A1 agonist, N6-(25)-[2-endo-norbonyl] adenosine [(S)-ENBA] produced a significant increase in tension at 1 pM, reaching 28% above control at 10 nM. The response to (S)-ENBA was also biphasic, with a fall in tension at 10 microM. The relatively non-specific agonist 5'-N-ethylcarboxamidoadenosine (NECA) produced a small, but significant, increase in tension at 1 microM, with no subsequent decline in tension at higher concentrations. These results allow us to assign a tentative structure-activity relationship (SAR) for an increase in tension of (S)-ENBA much much greater than R-PIA greater than or equal to 2-Cl ADO = N-6 CPA = NECA; for the decrease, the SAR is (S)-ENBA greater than 2-Cl ADO greater than R-PIA greater than N-6 CPA = NECA.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Chloroadenosine↗

Stage-dependency of apoptosis and the blood-testis barrier in the dogfish shark (Squalus acanthias): cadmium-induced changes as assessed by vital fluorescence techniques.

Naturally occurring heavy metals and synthetic compounds are potentially harmful for testicular function but evidence linking heavy metal exposure to reduced semen parameters is inconclusive. Elucidation of the exact stage at which the toxicant interferes with spermatogenesis is difficult because the various germ cell stages may have different sensitivities to any given toxicant, germ cell development is influenced by supporting testicular somatic cells and the presence of inter-Sertoli cell tight junctions create a blood-testis barrier, sequestering meiotic and postmeiotic germ cells in a special microenvironment. Sharks such as Squalus acanthias provide a suitable model for studying aspects of vertebrate spermatogenosis because of their unique features: spermatogenesis takes place within spermatocysts and relies mainly on Sertoli cells for somatic cell support; spermatocysts are linearly arranged in a maturational order across the diameter of the elongated testis; spermatocysts containing germ cells at different stages of development are topographically separated, resulting in visible zonation in testicular cross sections. We have used the vital dye acridine orange and a novel fluorescence staining technique to study this model to determine (1) the efficacy of these methods in assays of apoptosis and blood-testis barrier function, (2) the sensitivity of the various spermatogonial generations in Squalus to cadmium (as an illustrative spermatotoxicant) and (3) the way that cadmium might affect more mature spermatogenic stages and other physiological processes in the testis. Our results show that cadmium targets early spermatogenic stages, where it specifically activates a cell death program in susceptible (mature) spermatogonial clones, and negatively affects blood-testis barrier function. Since other parameters are relatively unaffected by cadmium, the effects of this toxicant on apoptosis are presumably process-specific and not attributable to general toxicity.

Animals↗

The sided action of Na+ and of K+ on reconstituted shark (Na+ + K+)-ATPase engaged in Na+-Na+ exchange accompanied by ATP hydrolysis. I. The ATP activation curve.

The ATP hydrolysis dependent Na+-Na+ exchange of reconstituted shark (Na+ + K+)-ATPase is electrogenic with a transport stoichiometry as for the Na+-K+ exchange, suggesting that translocation of extracellular Na+ is taking place via the same route as extracellular K+. The preparation thus offers an opportunity to compare the sided action of Na+ and K+ on the affinity for ATP in a reaction in which the intermediary steps in the overall reaction seems to be the same without and with K+. With Na+ but no K+ on the two sides of the enzyme, the ATP-activation curve is hyperbolic and the affinity for ATP is high. Extracellular K+ in concentrations of 50 microM (the lowest tested) and up gives biphasic ATP activation curves, with both a high- and a low-affinity component for ATP. Cytoplasmic K+ also gives biphasic ATP-activation curves, however, only when the K+ concentration is 50 mM or higher (Na+ + K+ = 130 mM). The different ATP-activation curves are explained from the Albers-Post scheme, in which there is an ATP-dependent and an ATP-independent deocclusion of E2(Na2+) and E2(K2+), respectively, and in which the dephosphorylation of E2-P is rate limiting in the presence of Na+ (but no K+) extracellular, whereas in the presence of extracellular K+ it is the deocclusion of E2(K2+) which is rate limiting.

Adenosine Triphosphate↗

Taurine and cell volume maintenance in the shark rectal gland: cellular fluxes and kinetics.

Tissue slices of shark rectal gland are studied to examine the kinetics of the cellular fluxes of taurine, a major intracellular osmolyte in this organ. Maintenance of high steady-state cell taurine (50 mM) is achieved by a ouabain-sensitive active Na+-dependent uptake process and a relatively slow efflux. Uptake kinetics are described by two saturable taurine transport components (high-affinity, Km 60 microM; and low-affinity, Km 9 mM). [14C]Taurine uptake is enhanced by external Cl-, inhibited by beta-alanine and unaffected by inhibitors of the Na+/K+/2Cl- co-transport system. Two cellular efflux components of taurine are documented. Incubation of slices in p-chloromercuribenzene sulfonate (1 mM) reduces taurine uptake, increases efflux of taurine and induces cell swelling. Studies of efflux in isotonic media with various cation and anion substitutions demonstrate that high-K+ markedly enhances taurine efflux irrespective of cell volume changes (i.e. membrane stretching is not involved). Moreover, iso-osmotic cell swelling induced in media containing propionate is not associated with enhanced efflux of taurine from the cells. It is suggested that external K+ exerts a specific effect on the cytoplasmic membrane to increase its permeability to taurine.

4-Chloromercuribenzenesulfonate↗

Variable stoichiometry in reconstituted shark Na,K-ATPase engaged in uncoupled efflux.

In liposomes with reconstituted shark Na,K-ATPase produced to contain no internal K+ or Na+ addition of external Na+ and ATP induce an uncoupled Na+ efflux on inside-out oriented pumps which is electrogenic and accompanied by hydrolysis of ATP (Cornelius, F. (1989) Biochem. Biophys. Res. Commun. 160, 801-807). At saturating cytoplasmic Na+ the net-charge translocated per ATP molecule split is compatible with a coupling ratio of Nacyt transported per ATP split of 3:1 at pH greater than or equal to 7.0. However, this ratio decreases to 1.5:1 below pH 7.0. At non-saturating cytoplasmic Na+ the 3:1 stoichiometry is attained at pH 7.0-7.5, whereas outside this range of pH the net-charge translocated per ATP molecule split decreases.

Adenosine Triphosphate↗

Uncoupled Na+-efflux on reconstituted shark Na,K-ATPase is electrogenic.

In liposomes with reconstituted shark Na,K-ATPase produced to contain sucrose addition of external Na+ and ATP induce an uncoupled Na+-efflux on inside-out oriented pumps which can be inhibited by digitoxigenin. This flux mode is found to be electrogenic and accompanied by hydrolysis of ATP. The coupling ratio of Nacyt transported per ATP split is 3:1 measured as the initial rate of rise in transmembrane potential and initial rate of liberated Pi.

Adenosine Triphosphate↗

Effects of mammalian ACTH on potential fuels and gluconeogenic substrates in the plasma of the spiny dogfish shark (Squalus acanthias).

The effects of mammalian ACTH on plasma beta-hydroxybutyrate, glucose, alanine, and lactate levels were determined in the unfed spiny dogfish shark (Squalus acanthias). Serial blood samples were collected from a cannula implanted in the dorsal aorta. The metabolite levels were estimated by standard enzymatic procedures for 4 days before treatment and for 4 days after treatment. Plasma glucose, alanine, and lactate, but not beta-hydroxybutyrate, declined after the surgery, but the metabolites were relatively stable for at least 48 hr before treatment. ACTH (40 units/kg) or the control solution were infused via the cannula after the morning blood sample on postoperative Day 4 and again after the 12-hr blood sample. Control plasma beta-hydroxybutyrate, glucose, and lactate did not change significantly, but alanine levels increased approximately 29% by 96 hr when sampling ended. Plasma beta-hydroxybutyrate levels were not significantly changed by ACTH treatment. Plasma glucose levels increased approximately 36% by 3 hr after ACTH infusion, remained elevated following the second ACTH treatment, and then declined to approximately the initial levels by 96 hr. Plasma alanine levels increased approximately 53% by 0.75 hr after ACTH treatment, were still approximately 15% greater than the initial levels by 12 hr, rose again after the second ACTH infusion at 12 hr, and then declined to near the control levels by 96 hr. Plasma lactate levels increased approximately 107% by 1.5 hr after ACTH infusion and then decreased to approximately 22% greater than the initial levels by 12 hr. Lactate levels increased after the second ACTH infusion and remained approximately 58% greater than the initial levels when sampling ended at 96 hr. The results indicate that the pituitary-adrenocortical axis hormones do not directly influence plasma beta-hydroxybutyrate levels. However, the data support the suggestion that the hyperglycemic action of the pituitary-adrenocortical axis hormones is mediated, at least in part, by the provision of alanine and lactate substrates for gluconeogenesis.

3-Hydroxybutyric Acid↗