Comparative study between the effects of hyaluronic acid and acid galactan purified from eggs of the mollusk Pomacea sp in wound healing

To compare the effect of hyaluronic acid (HA) and of AG on the healing of intestine wounds. Methods: The semi-purified extract of the eggs of the mollusc was obtained by fractionation with ammonium sulfate and purification for ion-exchange chromatography. The obtained galactans were eluted in water (neutral galactan) and in 0.1 and 0.2M NaCl (acidic galactans). The in vivo study was performed with 45 “Wistar” rats, separated in three groups (n=15). Solutions containing HA 1%, GA 1% or saline solution 0,9%, was placed topically on the sutures of wounds in the small intestine of the rats. After 05, 10 and 21 days the animals were sacrificed and biopsy of the healing tissue was done. Results: The hystologic grading was more significant for HA and AG groups when compared to the group C. AG stimulated the appearance of macrophages, giant cells and increase in the concentration of collagen in the area of the wound when compared to HA. Conclusion: The topical use of GA in intestinal wounds promoted the anticipation of events that are important in the wound healing

Estrutura e dinâmica do fitoplânctone bacterioplâncton em cultivos de camarão no Rio Grande do Norte-Brasil: impacto sobre o ambiente natural

This work aimed to study the structure and dynamic of Phytoplankton and Bacterioplankton in a complete cycle of shrimp cultivation (Litopenaeus vannamei) and determine the environmental factors responsible for the structural changes of these communities. The study was realized in a saltwater shrimp farm (Macaíba, RN), between September/2005 and February/2006, and in a freshwater shrimp farm (Ceará Mirim, RN), between May/2007 and September 2007. The samplings were collected weekly in saltwater farm and every fifteen days in freshwater farm. Total phosphorus, chlorophyll a and environmental parameters (pH, dissolved oxygen, salinity, temperature, depth and water transparency) were measured. Qualitative and quantitative analysis of the phytoplankton and bacterioplankton were carried out. The Shannon-Wiener ecologic indexes of diversity and the Pielou equitability indexes were calculated to the phytoplankton. Bacterial density was determined by epifluorescence microscopy. The data were statistically analyzed by Pearson correlation and t-Test. Chlorophycea were predominat in salt water and in the captation/drainage point (24 to 99%). Diatoms had higher wealth. The species Choricystis minor had the highest occurrence (100%) and dominance (90-100%), thus showing its adaptation to the high temperatures, salinity and low water transparency conditions. Filamentous Cyanobacteria like Oscillatoria sp., Pseudoanabaena sp. and Phormidium sp. had constant levels. The negative correlation between chlorophycea and water transparency, and the positive correlation between chlorophyll a and salinity, showed that the phytoplankton was well adapted to the low transparency and to the high salinity. The bacterioplankton was negatively correlated with the total phosphorus and salinity. In freshwater, Cyanobacteria were predominant (>80%), presenting some producers of toxins species like Microcystis sp., Aphanizomenon sp., Cylindrospermopsis raciborskii e Anabaena circinalis. Cyanobacterial density and total phosphorus and chlorophyll a concentrations exceeded the maximum value allowed by legislation. The means of total phosphorus varied from 264 to 627 Wg.L-1 and the means of chlorophyll a oscillated between 22 and 182 Wg.L-1. The phytoplankton species were selected by low availability of the light, high pH, temperature and high availability of total phosphorus. The bacterioplankton showed high densities (5,13 x 107 to 8,50 x107 Bac.mL-1). The studied environments (ponds and rivers) presented a high level of trophic state based on the high concentrations of chlorophyll a and total phosphorus and cyanobacteria dominance. The composition of species in the ponds and rivers was similar, as well as high concentrations of total phosphorus and chlorophyll a, highlighting the pollution caused by the discharges of the farms in natural environment

Avaliação da toxicidade de florações naturais e de cultura de cianobactérias: efeitos sobre Ceriodaphnia silvestri (Crustacea, Cladocera)

The incidence of toxic cyanobacterial blooms is one of the important consequences of eutrophication in aquatic ecosystems. It is a very common phenomenon in reservoirs and shrimp ponds in the State of Rio Grande do Norte (RN), Brazil. Cyanobacterias produce toxins which can affect aquatic organisms and men trough the food chain. Aiming to contribute to the studies of cyanobacterias in RN, we propose: a) to evaluate the toxicity of isolated cyanobacterias in important fresh-water environments; and b) to verify the effects of both natural and cultured blooms occurred in reservoirs for human supply and in the cladoceran Ceriodaphnia silvestrii. This study was carried out using samples of natural blooms occurred between March and October of 2004 in Gargalheiras Dam (08º L e 39º W), in July of 2004 in Armando Ribeiro Gonçalves Dam (06o S e 37o W) and in commercial shrimp ponds (Litopenaeus vannamei) located in fresh-water environments. The samples were collected with plankton net (20µm.) for identification, isolation and obtaining of phytoplanktonic biomass for liophilization and later toxicity bioassays. The toxicity of cultured samples and natural blooms was investigated through bioassays in Swiss mice. Quantification of cyanobacteria in samples was conducted following the Ütermol method, with 300mL samples fixed with lugol. The toxicity test with Ceriodaphnia silvestrii followed ABNT, 2001 recommendations, and were accomplished with natural hepatotoxic bloom s samples and cultured samples of both non-toxic and neurotoxic C. raciborskii. In this test, five newborns, aged between 6 and 24 hours, were exposed to different concentrations (0 a 800 mg.L-1) of crude cyanobacterial extracts during 24 and 48 hours. Three replicates were used per treatment. The pH, temperature and dissolved oxygen at the beginning and after 24 and 48hours from the test were measured. We estimated the CL50 through the Trimmed Spearman-Karber method. The blooms were constituted by Microcystis panniformis, M. aeruginosa, Anabaena circinalis, Cylindrospermopsis raciborskii and Planktothrix agardhii, producers of mycrocistin-LR confirmed with HPLC analysis. Samples of hepatotoxic blooms registered toxinogenic potential for C. silvestrii, with CL50-24h value of 47.48 mg.L-1 and CL5048h of 38.15 mg.L-1 for GARG samples in march/2005; CL50-24h of 113,13 mg.L-1 and CL5048h of 88,24 mg.L-1 for ARG July/2004; CL50-24h of 300.39 mg.L-1 and CL50-48h of 149.89 mg.L-1 for GARG October/2005. For cultured samples, values of CL50-24h and CL50-48h for C. raciborskii toxic strains were 228.05 and 120.28 mg.L-1, respectively. There was no mortality of C. silvestrii during the tests with non-toxic C. raciborskii strain. The toxicity test with C. silvestrii presented good sensitivity degree to cyanotoxins. The toxicity of natural hepatotoxic blooms samples (microcystins) and cultured neurotoxic saxitoxins producer samples analyzed in this study give us strong indications of that toxin s influence on the zooplanktonic community structure in tropical aquatic environments. Eleven cyanobacteria strains were isolated, representing 6 species: Anabaenopsis sp., Cylindrospermopsis raciborskii, Chroococcus sp., Microcystis panniformis, Geitlerinema unigranulatum e Planktothrix agardhii. None presented toxicity in Swiss mice. The strains were catalogued and deposited in the Laboratório de Ecologia e Toxicologia de Organismos Aquáticos (LETMA), in UFRN, and will be utilized in ecotoxicológical and ecophysiological studies, aiming to clarify the causes and control of cyanobacterial blooms in aquatic environments in RN. This state s reservoirs must receive broader attention from the authorities, considering the constant blooms occurring in waters used for human consumption

Purificação e caracterização de uma β-glucuronidase extraída do molusco mesogastropoda ampularidade Pomacea sp

This work studies the involved enzymatic way in the metabolism of glycosaminoglycans sulfateds in the mollusc Pomacea sp. Had been identified endoglycosidases and exoglycosidases in the enzymatic extract of the mollusc Pomacea sp by means of hydrolysis activity in condroitim sulphate of whale cartilage and of the p-Nitrofenil-β-glucuronide, respectively. The enzymatic extracts qere obtained of Pomacea sp. being used of 0.1 sodium acetate buffer, pH 5.0 and later centrifugated the 8,000 x g and the presents proteins in the sobrenadante were submitted to the fractionament with two crescents ammonium sulphate concentrations, the visualized activity biggest in the F2 fraction (50-80%). The β-glucuronidase (F3) was isolated in gel chromatography filtration Biogel 1.5m, the purification degree was ratified in Chromatography Liquid of high efficiency (HPLC). The enzyme was purificated 6.362,5 times with 35,6% yield. The β -glucuronidase isolated in this work showed a molecular mass of 100 kDa, determined for eletroforese in poliacrilamida gel . The determination of the ideal kinetic parameters for the catalysis of the p-nitrofenil- β -glucuronide for β-glucuronidase, showed excellent activity in pH 5,0 and temperature 65ºC for 6 hours and apparent Km of 72 x 10-2 mM. It is necessary for the total degradation of 3mM of p-N-β-glucoronide, the amount of 1,2μg of ss-glucuronidase. The BaCl2 increased the activity of ss-glucuronidase, and the activity was inhibited completely by the composites SDS and NaH2PO4

Caracterização e comparação de genes expressos em ápices meristemáticos de cana-de-açúcar cultivados em SP e RN

In this work, we used sugarcane as a model due to its importance for sugar and ethanol production. Unlike the current plant models, sugarcane presents a complex genetics and an enormous allelic variation. Here, we report the analysis of SAGE libraries produced using the shoot apical meristem from contrasted genotypes by flowering induction (non-flowering vs. early-flowering varieties) grown under São Paulo state conditions. The expression pattern was analyzed using samples from São Paulo (SP) and Rio Grande do Norte (RN) states. These results showed that cDNAs identified by SAGE libraries had differential expression only in São Paulo state samples. Furthermore, the cDNA identified CYP (Citocrome P450) was chosen for in silico and genome characterization because it was found in SAGE libraries and subtractive libraries from samples from RN. Phylogenetic trees showed the relationship for these sequences. Furthermore, the qRT-PCR for CYP showed a potential role as flowering indutor for RN samples considering different isophorms. Considering the results present here, it can be consider that CYP gene may be used as molecular marker

Estudo comparativo da variação temporal da comunidade fitoplanctônica do açude Cruzeta inserido no bioma caatinga, Rio Grande do Norte

The Cruzeta impoundment, situated in the city of Cruzeta, hinterland of the state of the Rio Grande do Norte state has significant importance to the municipality as it represents the only source of supplying water to the region. It was hypothesized that the regional consequence of the global warming and the warming of surface water could substantially contribute the significant growth of the aquatic macrophytes in the years 2008-2009. The growth of these vegetation believed to be improved the degree of water transparency and as a consequence of the improved growth of phytoplankton species and chlorophyll biomass. At the same time the aquatic macrophyte could interact and compete potentially for the dissolved inorganic nutrients resources and the phytoplankton community. This study presents a comparison of years 2004-2005 when it did not have the expressive presence of the aquatic macrophyte community or restricted to the littoral region. In contrast, the years 2008-2009 showed a significant growth of the aquatic macrophyte in the Cruzeta/RN impoundments. The present study is an attempt to elucidate the significant presence of the aquatic macrophyte, Eichhornia crassipes, Ceratophyllum submersum, Nymphea sp and Pistia sp, and its interference on the ecology of phytoplankton. The samplings had been carried out from September of 2008 to April of 2009 and consistently between 10:00 h and 12:00 h with the aid of Van Dorn bottle and the plankton net of mesh size 20 Qm. The collections were made in three depths ie., surface, mid-column and bottom. The Physical-Chemical parameters such as pH, temperature, electrical conductivity and dissolved oxygen had been analyzed in situ. The samples for analysis of nutrients and chlorophyll were kept under refrigeration for posterior analysis in the laboratory. Phytoplankton samples were preserved in Lugol-iodine and kept for sedimentation for quali-quantitative analysis of phytoplankton. Enumeration of cells, colonies and filaments was done with the aid of Sedgwick-Rafter counting chamber and expressed as numbers/ml. Chlorophyll a was analyzed as a functional component of phytoplankton biomass and extracted with cold 90% acetone. The results indicate that the chlorophyll concentration varied between 5,65-8,08 Qg.L-1 for the dry period and 5,09-6,23 Qg.L-1 for the rainy period and showed considerable reduction when compared to the values to the 2004-2005 study period. The temperature was always presented higher in relation to the 2004-2005 study. Phytoplankton species showed a relative abundance of the Cyanophyceae for both the period of dry and rainy. The predominance species are filamentous Leptolymbya geophila Borzi (Planctolyngbya sp), Anabaena plankctônica Brunnthaler, Oscillatória limosa Ag. and Cylindrospermopsis raciborskii (Wolosz). The concentration of the nutrients such as nitrate and orto-phosfato had always presented higher values during the rainy period and the ammoniacal nitrogen retained moderate values in the dry period and a slight increase in rainy season. The main conclusions are the reduction of the concentration of chlorophyll, diversity of phytoplankton, and the increase in temperature and transparency of the water during the period of the study

Avaliação da composição química, atividade antioxidante, antibacteriana, antinoceptiva, antiinflamatória e toxicidade do extrato metanólico e frações de folhas de Spondias sp. (Anacardiaceae)

Spondias sp. (Anacardiaceae), popularly known as cajá-umbu, is an endemic plant from Northeastern Brazil, where their leaves are widely used in folk medicine to treat inflammatory processes, while their fruits have a great agro industrial potential. This study was designed to evaluate hepatoprotective, antinociceptive, antioxidant, antimicrobial and anti-inflammatory properties, as well as the acute toxicity and repeated dose 28, using a methanolic extract (MES), a fraction rich in flavonoids (FRF) and a precipitate from Spondias sp.leaves. The antioxidant activity of them was valued to evaluate their free radical scavenger capacity by DPPH test, whereas MES and FRF were used to evaluate while the preventive action on carbon tetrachloride (CCl4)-induced hepatotoxicity. Seven groups (n=5) of female Wistar rats were used as follows: control group, CCl4-intoxicated group treated with EMS (500 mg/kg) for 7 days, three CCl4-intoxicated groups treated with FRF (25, 50 and 75 mg/kg) for 7 days and the CCl4-intoxicated group treated with Legalon ® (silimarina; (phytotherapeutic reference) (50 mg/kg; 7 days). MES and FRF showed a protective action against liver injury induced by CCl4, being observed a significant reduction of serum enzyme activity marker of liver damage (alanine transaminase and aspartate transaminase). On the other hand, the lipid peroxidation (SRAT) decrease, as well as the increase of glutathione content and enzyme activity of antioxidant defense system (SOD, CAT, GPx) toward near normal values indicated the ability of EMS to restore the oxidative imbalance induced by CCl4. The histological analysis confirmed the hepatoprotection, compared to degenerative changes in CCl4-treated group. This hepatoprotetor effect was similar to that shown by Legalon®. The in vitro high antioxidant capacity of extract (93.16 ± 1.00%) showed analogous results to those obtained by Carduus marianus BHT (reference standard). This fact explains the obtained results in vivo. Although no antimicrobial activity was detected, EMS and FRF promoted the antinociceptive effect induced in the second phase by the intraplantar formalin test, evidencing the anti-inflammatory action; confirmed by the carrageenan-induced peritonitis model. The evaluation of the mechanical allodynia (CFA a 80%) demonstrated the involvement of the Spondias sp. chemical composition in the anti-inflammatory activity toward the acute processes. The acute exposure and repeated dose during 28 days did not produce significant changes in the parameters that evaluate toxicity. Together the experimental results reveal, that Spondias sp. leaf extracts have a promising potential in pharmaceutical area, and due to its non-toxic condition present efficiency and security

Biossorção utilizando alga marinha (sargassum sp.) aplicada em meio orgânico

In this work, biosorption process was used to remove heavy metals from used automotive lubricating oils by a bus fleet from Natal-RN-Brazil. This oil was characterized to determine the physical-chemistry properties. It was also characterized the used oil with the aim of determining and quantifying the heavy metal concentration. Fe and Cu were the metals existent in large concentration and these metals were choused to be studied in solubilization process. For the biosorption process was used the seaweed Sargassum sp for the study of influencing of the metals presents separately and with other metals. It was also studied the effect of the protonation treatment of alga with the objective to know the best efficiency of heavy metals removal. The study of the solubilization showed that the presence of more than a metal favors the solubilization of the metals presents in the oil and consequently, it favors the biosorption process, what becomes interesting the perspective application in the heavy metals removal in lubricating oils used, because the presence of more than a heavy metal favors the solubility of all metals present. It was observed that the iron and copper metals, which are present in large concentration, the protonated biosorbtent was more effective. In this study we used as biomass the marine alga Sargassum sp to study the influence of agitation velocity, temperature and initial biomass concentration on the removal of iron and copper from used lubricant oils. We performed an experimental design and a kinetic study. The experiments were carried out with samples of used lubricant oil and predetermined amounts of algae, allowing sufficient time for the mixture to obtain equilibrium under controlled conditions. The results showed that, under the conditions studied, the larger the amount of biomass present, the lower the adsorption capacity of the iron and of the copper, likely due to a decrease in interface contact area. The experimental design led us to conclude that a function can be obtained that shows the degree of influence of each one of the system variables