Anticancer Activity of Pasak Bumi Root Extract (Eurycoma longifolia Jack) on Raji Cells.

<b>Background and Objective:</b> The use of the roots of the pasak bumi (<i>E. longifolia</i> Jack) to treat cancer has been studied widely, however, the scientific basis of these plants used as an anticancer drug is widely unknown. The purpose of this study was to examine the anticancer activity of ethyl acetate and non-ethyl acetate fractions of pasak bumi roots in Raji cells. <b>Materials and Methods:</b> The cytotoxicity test is using the direct cell count method with trypan blue staining. The growth inhibition is using doubling time analysis of Raji cells. Observation of the apoptotic events of Raji cells used ethidium bromide staining, while observing the expression of p53 protein in Raji cells was done by immunohistochemical staining. <b>Results:</b> The results of the cytotoxicity and doubling time test showed that the activity of the non-ethyl acetate fraction was greater than that of the roots of pasak bumi. The lower concentration of non-ethyl acetate fraction of pasak bumi roots was able to delay the multiplication time of Raji cells which was greater than that of ethyl acetate. The results of the cytotoxicity and doubling time test showed that the activity of the non-ethyl acetate fraction was greater than that of the roots of pasak bumi. <b>Conclusion:</b> It can be concluded that the ethyl acetate and non-ethyl acetate fractions of the roots of pasak bumi have cytotoxic and antiproliferative activity on Raji cells, however they cannot induce apoptosis in Raji cells. The death of Raji cells is through the mechanism of inhibiting Raji cell proliferation as evidenced by an increase in p53 protein expression.

Electrosensory Impairment in the Atlantic Stingray, Hypanus sabinus, After Crude Oil Exposure.

Elasmobranchs are renowned for their extremely sensitive electrosensory system, which is used to detect predators, prey, and mates, and is possibly used for navigation. The proper functioning of the electrosensory system is thus critical to fitness. The objective of this study was to test whether exposure to crude oil impairs the electroreceptive capabilities of elasmobranch fishes. Electrosensory function was quantified from six stingrays before and after exposure to a concentration of oil that mimicked empirically measured concentrations along the coast of Louisiana following the Deepwater Horizon spill. Orientation distance (cm), and angle with respect to the dipole axis of a prey-simulating electric field were used to derive the electric field intensity that elicited a response. Oil exposed stingrays continued to exhibit feeding behavior, but they initiated orientations to prey-simulating electric fields from a significantly closer orientation distance. The mean orientation distance after oil exposure was 5.29 ± 0.41 SE cm compared to a pre-exposure orientation distance of 7.16 ± 0.66 SE cm. Stingrays required a mean electric field intensity of 0.596 ± 0.21 SE µV cm-1 to initiate a response after oil exposure, compared to a mean of only 0.127 ± 0.03 SE µV cm-1 in uncontaminated seawater. Oil exposed stingrays thus exhibited a response to a stimulus approximately 4.7 times greater than controls. Stingrays impacted by an oil spill appear to experience reduced electrosensory capabilities, which could detrimentally impact fitness. This study is the first to quantify the effects of crude oil on behavioral electrosensory function.

Popularidade e uso medicinal do pinhão-roxo (Jatropha gossipifolia L. ) em comunidades ribeirinhas do pantanal do Mato Grosso do Sul, com ênfase em envenenamentos causados por arraias fluviais / Popularity and medicinal use of purple pine nut (Jatropha gossipifolia L.) in riverside communities in the Pantanal of Mato Grosso do Sul, with an emphasis on poisonings caused by river stingrays

O Pantanal é a maior planície inundável do planeta. A população humana da região utiliza vegetais que encontram no dia a dia e, com o tempo, isso os levou a descobrir diferentes finalidades alimentares e até usos terapêuticos dessas plantas. Este estudo tem como objetivo compreender a popularidade do pinhão-roxo (Jatropha gossipifolia L.) e seu uso como planta medicinal em duas comunidades ribeirinhas do município de Corumbá (MS), Brasil, com ênfase nos envenenamentos causados por arraias de água doce, frequentes na área. Mais da metade dos entrevistados utilizou o pinhão-roxo em várias doenças, especialmente no controle de sangramentos e na cicatrização de feridas. O uso de infusões em úlceras crônicas causadas por peixes peçonhentos é amplamente mencionado pelos pescadores entrevistados, particularmente em envenenamentos com necrose cutânea causada por arraias de água doce. Outras indicações citadas são o tratamento de micoses cutâneas e odontalgias, que não são mencionadas nas referências anteriores, indicando a necessidade de estudos clínicos e farmacológicos adicionais para avaliar esses propósitos.

Effects of collagen peptides from skate (Raja kenojei) skin on improvements of the insulin signaling pathway via attenuation of oxidative stress and inflammation.

It has been well established that hepatic insulin signaling is significantly affected by the antioxidative status of the liver. In this study, we first confirmed that skate skin collagen peptide (SSCP) administration has dose-dependent positive effects on the change in the glucose level as evidenced by oral glucose tolerance tests. Therefore, the beneficial effects of SSCP-showing antioxidative and anti-inflammatory activities-on insulin resistance were examined in high-fat diet (HFD)-fed mice. C57BL/6J mice orally received SSCP at doses of 100, 200, and 300 mg per kg bw per day along with a HFD for 8 weeks (n = 9 per group). Water was given to the HFD- or chow diet-only group as a vehicle. Compared with the HFD group, the final body weight was reduced in all the SSCP-treated groups in a dose-dependent manner. The hepatic protein expression levels of the phosphorylated insulin receptor substrate, phosphorylated phosphatidylinositol 3-kinase, and phosphorylated protein kinase B were increased in the SSCP-treated groups, which led to reduced plasma insulin and HOMA-IR levels (P < 0.05). The hepatic protein expression levels of nuclear factor erythroid 2-related factor 2-mediated antioxidant enzymes were increased in the SSCP-treated groups, whereas those of nuclear factor kappa B-regulated inflammatory enzymes and mediators were decreased (P < 0.05). These effects were dose-dependent. It is apparent that SSCP might enhance insulin sensitivity by increasing the antioxidative status and suppressing the inflammatory response in the liver.

Impact of Prevalence Ratios of Chondroitin Sulfate (CS)- 4 and -6 Isomers Derived from Marine Sources in Cell Proliferation and Chondrogenic Differentiation Processes.

Osteoarthritis is the most prevalent rheumatic disease. During disease progression, differences have been described in the prevalence of chondroitin sulfate (CS) isomers. Marine derived-CS present a higher proportion of the 6S isomer, offering therapeutic potential. Accordingly, we evaluated the effect of exogenous supplementation of CS, derived from the small spotted catshark (Scyliorhinus canicula), blue shark (Prionace glauca), thornback skate (Raja clavata) and bovine CS (reference), on the proliferation of osteochondral cell lines (MG-63 and T/C-28a2) and the chondrogenic differentiation of mesenchymal stromal cells (MSCs). MG-G3 proliferation was comparable between R. clavata (CS-6 intermediate ratio) and bovine CS (CS-4 enrichment), for concentrations below 0.5 mg/mL, defined as a toxicity threshold. T/C-28a2 proliferation was significantly improved by intermediate ratios of CS-6 and -4 isomers (S. canicula and R. clavata). A dose-dependent response was observed for S. canicula (200 µg/mL vs 50 and 10 µg/mL) and bovine CS (200 and 100 µg/mL vs 10 µg/mL). CS sulfation patterns discretely affected MSCs chondrogenesis; even though S. canicula and R. clavata CS up-regulated chondrogenic markers expression (aggrecan and collagen type II) these were not statistically significant. We demonstrate that intermediate values of CS-4 and -6 isomers improve cell proliferation and offer potential for chondrogenic promotion, although more studies are needed to elucidate its mechanism of action.

Recognizing peripheral ecosystems in marine protected areas: A case study of golden jellyfish lakes in Raja Ampat, Indonesia.

Peripheral marine ecosystems can harbor endemic diversity and attract tourism attention, yet are generally not included in conservation management plans due to their remoteness or inland positioning. A case study in Raja Ampat of seven landlocked marine lakes containing golden jellyfish (Mastigias spp.) was conducted to address the lack of fundamental insights into evolutionary, ecological and social contexts of these ecosystems. An interdisciplinary approach was taken towards identifying the jellyfish lakes as distinct management units in order to incorporate them into existing Marine Protected Areas. Mastigias papua populations showed strong genetic (ϕST: 0.30-0.86) and morphological (F = 28.62, p-value = 0.001) structure among lakes, with putative new subspecies. Risks arising from rapid increase in tourism to Raja Ampat (30-fold since 2007) warrant restrictions on jellyfish lake use. Recommendations are provided for adaptive management and science-based conservation policies for jellyfish lakes across Indonesia.

Effect of Deepwater Horizon Crude Oil Water Accommodated Fraction on Olfactory Function in the Atlantic Stingray, Hypanus sabinus.

The Deepwater Horizon oil spill was the largest accidental marine oil spill in history, releasing nearly 5 million barrels of crude oil. Crude oil causes both lethal and sublethal effects on marine organisms, and sensory systems have the potential to be strongly affected. Marine fishes rely upon the effective functioning of their sensory systems for detection of prey, mates, and predators. However, despite the obvious importance of sensory systems, the impact of crude oil exposure upon sensory function remains largely unexplored. Here we show that olfactory organ responses to amino acids are significantly depressed in oil exposed stingrays. We found that the response magnitude of the electro-olfactogram (EOG) to 1 mM amino acids decreased by an average of 45.8% after 48 h of exposure to an oil concentration replicating that measured in coastal areas. Additionally, in oil exposed individuals, the EOG response onset was significantly slower, and the clearing time was protracted. This study is the first to employ an electrophysiological assay to demonstrate crude oil impairment of the olfactory system in a marine fish. We show that stingrays inhabiting an area impacted by an oil spill experience reduced olfactory function, which would detrimentally impact fitness, could lead to premature death, and could cause additional cascading effects through lower trophic levels.

Anti-Obesity Effects of Collagen Peptide Derived from Skate (Raja kenojei) Skin Through Regulation of Lipid Metabolism.

This study investigated the anti-obesity effects of collagen peptide derived from skate skin on lipid metabolism in high-fat diet (HFD)-fed mice. All C57BL6/J male mice were fed a HFD with 60% kcal fat except for mice in the normal group which were fed a chow diet. The collagen-fed groups received collagen peptide (1050 Da) orally (100, 200, or 300 mg/kg body weight per day) by gavage, whereas the normal and control groups were given water (n = 9 per group). The body weight gain and visceral adipose tissue weight were lower in the collagen-fed groups than in the control group (p < 0.05). Plasma and hepatic lipid levels were significantly reduced by downregulating the hepatic protein expression levels for fatty acid synthesis (sterol regulatory element binding protein-1 (SREBP-1), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC)) and cholesterol synthesis (SREBP-2 and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR)) and upregulating those for ß-oxidation (peroxisome proliferator-activated receptor alpha (PPAR-α) and carnitine palmitoyltransferase 1 (CPT1)) and synthesis of bile acid (cytochrome P450 family 7 subfamily A member 1 (CYP7A1)) (p < 0.05). In the collagen-fed groups, the hepatic protein expression level of phosphorylated 5' adenosine monophosphate-activated protein kinase (p-AMPK) and plasma adiponectin levels were higher, and the leptin level was lower (p < 0.05). Histological analysis revealed that collagen treatment suppressed hepatic lipid accumulation and reduced the lipid droplet size in the adipose tissue. These effects were increased in a dose-dependent manner. The findings indicated that skate collagen peptide has anti-obesity effects through suppression of fat accumulation and regulation of lipid metabolism.

Selenium and Mercury Interactions in Apex Predators from the Gulf of Trieste (Northern Adriatic Sea).

Since the environmental levels of selenium (Se) can moderate the bioaccumulation and toxicity of mercury (Hg) in marine organisms, their interactions were studied in seawater, sediments, plankton and the benthic (Bull ray Pteromylaeus bovinus, Eagle ray Myliobatis aquila) and the pelagic (Pelagic stingray Dasyiatis violacea) rays, as apex predators in the Gulf of Trieste (Northern Adriatic Sea). Male and female rays showed no difference in the Se contents in muscle tissue. Pelagic species contained higher Se levels in muscle but slightly lower levels in the livers of both genders. The Hg/Se ratios in seawater dissolved and colloidal fractions, plankton and sediment were <0.5, while those in particulate matter were <1.3. In benthic ray species, a parallel increase in Se and Hg in muscle was observed, so that an increased in Hg (MeHg) bioaccumulation results in Se coaccumulation. The Hg/Se ratios (molar) in muscle and liver of pelagic and benthic rays were <1.4 and <0.7, respectively. The low levels of Hg in muscle and liver in all the ray species corresponded to low Hg/Se ratios and increases in muscle and liver to 1 at 7 µg/g, dry weight (dw) and 5 µg/g dw, respectively, i.e., about 1.6 µg/g wet weight (ww).

Local-regional anesthesia in the management of stingray stings: Experience of the Bouffard medical-surgical hospital in Djibouti.

Stingray injuries are very painful. Systemic analgesics are ineffective, and the use of local-regional anesthesia has been reported. This retrospective descriptive study reviewed all cases of stingray injuries seen at the emergency department of the Bouffard Hospital (Djbouti, Africa) between 2011 and 2014. The study included 35 patients. Most of the injuries (n= 31, 89%) concerned the lower limbs. Median pain intensity was 6 [5-8] on a visual analog scale of 0 (no pain) to 10. The following systemic medications were administered: acetaminophen to 13 (27%) patients, morphine to 8 (23%), and tramadol to 6 (17%). In all, 25 (71%) patients received local-regional anesthesia, 15 (60%) by injections at the ankle. All procedures were successful, and no adverse event was reported. This study reports clinical data about stingray injuries in the Red Sea area and highlights the interest of local-regional anesthesia in their management. Most of the procedures were distal and could be performed by trained emergency physicians.

Batoid locomotion: effects of speed on pectoral fin deformation in the little skate, Leucoraja erinacea.

Most batoids have a unique swimming mode in which thrust is generated by either oscillating or undulating expanded pectoral fins that form a disc. Only one previous study of the freshwater stingray has quantified three-dimensional motions of the wing, and no comparable data are available for marine batoid species that may differ considerably in their mode of locomotion. Here, we investigate three-dimensional kinematics of the pectoral wing of the little skate, Leucoraja erinacea, swimming steadily at two speeds [1 and 2 body lengths (BL) s-1]. We measured the motion of nine points in three dimensions during wing oscillation and determined that there are significant differences in movement amplitude among wing locations, as well as significant differences as speed increases in body angle, wing beat frequency and speed of the traveling wave on the wing. In addition, we analyzed differences in wing curvature with swimming speed. At 1 BL s-1, the pectoral wing is convex in shape during the downstroke along the medio-lateral fin midline, but at 2 BL s-1 the pectoral fin at this location cups into the flow, indicating active curvature control and fin stiffening. Wing kinematics of the little skate differed considerably from previous work on the freshwater stingray, which does not show active cupping of the whole fin on the downstroke.

Anticancer Activity of a Hexapeptide from Skate (Raja porosa) Cartilage Protein Hydrolysate in HeLa Cells.

In this study, the hexapeptide Phe-Ile-Met-Gly-Pro-Tyr (FIMGPY), which has a molecular weight of 726.9 Da, was separated from skate (Raja porosa) cartilage protein hydrolysate using ultrafiltration and chromatographic methods, and its anticancer activity was evaluated in HeLa cells. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay indicated that FIMGPY exhibited high, dose-dependent anti-proliferation activities in HeLa cells with an IC50 of 4.81 mg/mL. Acridine orange/ethidium bromide (AO/EB) fluorescence staining and flow cytometry methods confirmed that FIMGPY could inhibit HeLa cell proliferation by inducing apoptosis. Western blot assay revealed that the Bax/Bcl-2 ratio and relative intensity of caspase-3 in HeLa cells treated with 7-mg/mL FIMGPY were 2.63 and 1.83, respectively, significantly higher than those of the blank control (p < 0.01). Thus, FIMGPY could induce apoptosis by upregulating the Bax/Bcl-2 ratio and caspase-3 activation. Using a DNA ladder method further confirmed that the anti-proliferation activity of FIMGPY was attributable to its role in inducing apoptosis. These results suggest that FIMGPY from skate cartilage protein hydrolysate may have applications as functional foods and nutraceuticals for the treatment and prevention of cancer.

Injuries caused by aquatic animals in Brazil: an analysis of the data present in the information system for notifiable diseases.

INTRODUCTION: We present a review of injuries in humans caused by aquatic animals in Brazil using the Information System for Notifiable Diseases [ Sistema de Informação de Agravos de Notificação (SINAN)] database. METHODS: A descriptive and retrospective epidemiological study was conducted from 2007 to 2013. RESULTS: A total of 4,118 accidents were recorded. Of these accidents, 88.7% (3,651) were caused by venomous species, and 11.3% (467) were caused by poisonous, traumatic or unidentified aquatic animals. Most of the events were injuries by stingrays (69%) and jellyfish (13.1%). The North region was responsible for the majority of reports (66.2%), with a significant emphasis on accidents caused by freshwater stingrays (92.2% or 2,317 cases). In the South region, the region with the second highest number of records (15.7%), jellyfish caused the majority of accidents (83.7% or 452 cases). The Northeastern region, with 12.5% of the records, was notable because almost all accidents were caused by toadfish (95.6% or 174 cases). CONCLUSIONS: Although a comparison of different databases has not been performed, the data presented in this study, compared to local and regional surveys, raises the hypothesis of underreporting of accidents. As the SINAN is the official system for the notification of accidents by venomous animals in Brazil, it is imperative that its operation be reviewed and improved, given that effective measures to prevent accidents by venomous animals depend on a reliable database and the ability to accurately report the true conditions.

Injuries caused by aquatic animals in Brazil: an analysis of the data present in the information system for notifiable diseases

We present a review of injuries in humans caused by aquatic animals in Brazil using the Information System for Notifiable Diseases [ Sistema de Informação de Agravos de Notificação (SINAN)] database.

Feeding biomechanics of the cownose ray, Rhinoptera bonasus, over ontogeny.

Growth affects the performance of structure, so the pattern of growth must influence the role of a structure and an organism. Because animal performance is linked to morphological specialization, ontogenetic change in size may influence an organism's biological role. High bite force generation is presumably selected for in durophagous taxa. Therefore, these animals provide an excellent study system for investigating biomechanical consequences of growth on performance. An ontogenetic series of 27 cownose rays (Rhinoptera bonasus) were dissected in order to develop a biomechanical model of the feeding mechanism, which was then compared with bite forces measured from live rays. Mechanical advantage of the feeding apparatus was generally conserved throughout ontogeny, while an increase in the mass and cross-sectional area of the jaw adductors resulted in allometric gains in bite force generation. Of primary importance to forceful biting in this taxon is the use of a fibrocartilaginous tendon associated with the insertion of the primary jaw adductor division. This tendon may serve to redirect muscle forces anteriorly, transmitting them within the plane of biting. Measured bite forces obtained through electrostimulation of the jaw adductors in live rays were higher than predicted, possibly due to differences in specific tension of actual batoid muscle and that used in the model. Mass-specific bite forces in these rays are the highest recorded for elasmobranchs. Cownose rays exemplify a species that, through allometric growth of bite performance and morphological novelties, have expanded their ecological performance over ontogeny.

Pharmacological investigation and spectral characterization of bioactive compounds from crude extracts of sting ray, Dasyatis jenkinsii (Annandale, 1909).

AIM: Dasyatis jenkinsii is used traditionally to treat inflammatory complaints and arthritis by the fisherman community and local population. The present study was designed to scientifically investigate the traditional practice through the analgesic, anti-inflammatory, and organ toxicity studies and characterization of bioactive compounds of crude extracts of D. jenkinsii. METHOD: Solvent extract of homogenized fresh fish was prepared using petroleum ether and diethyl ether. The chemical and spectral analyses of extracts were carried out using FT-IR and GC-MS. Analgesic and anti-inflammatory activities were assessed by hot plate, tail clip, and carrageenan induced rat paw edema methods. The organ toxicity of each extract was assessed on brain, liver, and kidney of mice. RESULTS: The IR spectrum indicated the presence of aromatic and aliphatic compounds. GC-MS analysis revealed the presence of 1-(4-carboxy)phenylnona-2, 5-diene and 3-hydroxymono-glyceryl hydrogen phthalate in the petroleum ether extract and carboxyl serine, dihydrotryptophan, and indolyl carboxylic acid in the diethyl ether extract. Both extracts showed significant analgesic and anti-inflammatory effects in all the methods tested. The vital organs of the test animals were not affected by the crude extracts significantly. CONCLUSIONS: The presence of biologically active compounds in the crude extracts with analgesic and anti-inflammatory properties justifies the traditional knowledge and paves the way for isolation of these compounds for further experimentation.

Injuries by marine and freshwater stingrays: history, clinical aspects of the envenomations and current status of a neglected problem in Brazil

Stingrays are a group of rays - cartilaginous fish related to sharks - that have whiplike tails with barbed, usually venomous spines and are found around the world, especially the marine species. Despite recent reports of accidents involving these fish, they are not aggressive, reacting only when stepped on or improperly handled. Injuries by stingrays are seldom mentioned by historians, although they have always been present in riverine communities of inland waters and in South American coasts. Indeed, envenomations by stingrays are quite common in freshwater and marine fishing communities. Although having high morbidity, such injuries are neglected because they have low lethality and usually occur in remote areas, which favor the use of folk remedies. In the present review article, historical aspects of injuries caused by stingrays in Brazil and their distribution on the coast of São Paulo state and riverine communities of the North, Midwest and Southeast regions were studied. In addition, other aspects were analyzed such as clinical features, therapeutic methods, preventive measures and trends in occurrence of these accidents in the country, particularly in areas in which freshwater stingrays had not been previously registered, being introduced after breaching of natural barriers.(AU)

Injuries by marine and freshwater stingrays: history, clinical aspects of the envenomations and current status of a neglected problem in Brazil

Stingrays are a group of rays – cartilaginous fish related to sharks – that have whiplike tails with barbed, usually venomous spines and are found around the world, especially the marine species. Despite recent reports of accidents involving these fish, they are not aggressive, reacting only when stepped on or improperly handled. Injuries by stingrays are seldom mentioned by historians, although they have always been present in riverine communities of inland waters and in South American coasts. Indeed, envenomations by stingrays are quite common in freshwater and marine fishing communities. Although having high morbidity, such injuries are neglected because they have low lethality and usually occur in remote areas, which favor the use of folk remedies. In the present review article, historical aspects of injuries caused by stingrays in Brazil and their distribution on the coast of São Paulo state and riverine communities of the North, Midwest and Southeast regions were studied. In addition, other aspects were analyzed such as clinical features, therapeutic methods, preventive measures and trends in occurrence of these accidents in the country, particularly in areas in which freshwater stingrays had not been previously registered, being introduced after breaching of natural barriers.

Injuries by marine and freshwater stingrays: history, clinical aspects of the envenomations and current status of a neglected problem in Brazil

Stingrays are a group of rays – cartilaginous fish related to sharks – that have whiplike tails with barbed, usually venomous spines and are found around the world, especially the marine species. Despite recent reports of accidents involving these fish, they are not aggressive, reacting only when stepped on or improperly handled. Injuries by stingrays are seldom mentioned by historians, although they have always been present in riverine communities of inland waters and in South American coasts. Indeed, envenomations by stingrays are quite common in freshwater and marine fishing communities. Although having high morbidity, such injuries are neglected because they have low lethality and usually occur in remote areas, which favor the use of folk remedies. In the present review article, historical aspects of injuries caused by stingrays in Brazil and their distribution on the coast of São Paulo state and riverine communities of the North, Midwest and Southeast regions were studied. In addition, other aspects were analyzed such as clinical features, therapeutic methods, preventive measures and trends in occurrence of these accidents in the country, particularly in areas in which freshwater stingrays had not been previously registered, being introduced after breaching of natural barriers.