In vitro screening of Amazonian plants for hemolytic activity and inhibition of platelet aggregation in human blood

In the present study, different aerial parts from twelve Amazonian plant species found in the National Institute for Amazon Research's (INPA's) Adolpho Ducke Forest Reserve (in Manaus, Amazonas, Brazil) were collected. Separate portions of dried, ground plant materials were extracted with water (by infusion), methanol and chloroform (by continuous liquid-solid extraction) and solvents were removed first by rotary evaporation, and finally by freeze-drying which yielded a total of seventy-one freeze-dried extracts for evaluation. These extracts were evaluated initially at concentrations of 500 and 100 µg/mL for in vitro hemolytic activity and in vitro inhibition of platelet aggregation in human blood, respectively. Sixteen extracts (23 % of all extracts tested, 42 % of all plant species), representing the following plants: Chaunochiton kappleri (Olacaceae), Diclinanona calycina (Annonaceae), Paypayrola grandiflora (Violaceae), Pleurisanthes parviflora (Icacinaceae), Sarcaulus brasiliensis (Sapotaceae), exhibited significant inhibitory activity towards human platelet aggregation. A group of extracts with antiplatelet aggregation activity having no in vitro hemolytic activity has therefore been identified. Three extracts (4 %), all derived from Elaeoluma nuda (Sapotaceae), exhibited hemolytic activity. None of the plant species in this study has known use in traditional medicine. So, these data serve as a baseline or minimum of antiplatelet and hemolytic activities (and potential usefulness) of non-medicinal plants from the Amazon forest. Finally, in general, these are the first data on hemolytic and inhibitory activity on platelet aggregation for the genera which these plant species represent.

Comparação da soja germinada, com a soja em grão e com o farelo de soja na alimentação de suínos em crescimento

Three groups of 6 pigs, three months old, were fed the same basal ration of corn and mineral mixture ad libitum. The control group received soybeans oil meal (solvent proc.), the second group raw soybeans and the third one, sprouted soybeans. The feed intake, daily gain and conversion were practically the same in the three groups as the analisis of variance revealed. Conclusion is it does not pay to sprout soybeans for pigs.

Sôbre uma modificação na determinação do Molibdênio em Plantas

The author proposed a new modification on the thiocyanate-stannous chloride method for the determination of molibdenum, when is used a heavier-than-water solvent for extrating the colored molybdenum thiocyanate complex. Carbon tetrachloride - butyl alcohol is the mixture proposed, and the results obtained give a good precision and more sensibility than the other method that use carbon tetrachloride-isoamyl alcohol as extractant.

Lactacidemia: métodos de dosagem do ácido lático

A critical study of three methods for the determination of lactic acid (EDWARDS, MENDEL & GOLDSCHEIDER, MILLER & MUNTZ) is presented and some modifications are proposed. It was shown t hat more accurate results could be obtained with Edward's technic when an Iena glass filter is connected with the absorption tube. Before the dropping of the permanganate solution it is necessary to pass a current of air through the reaction flask to avoid the oxidation of the non-lactic acid substances which interfere with the reaction. The absorption tube must be maintained at 18°C during the destillation and the titration of the bisulphite binding aldehyde at 4°C. When the sample contains more than 5 mg it is useful to work with greater quantities of the bisulphite. More permanganate is consumed when the lactic acid concentration is higher. The sensivity of the method permits the titration of 0.04 mg to 5 mg of lactic acid in the sample. The calculated error of the method gave 0.018 % and the normal values for blood determined in 20 human cases averaged 10.30 mg per 100 ml (Table VI). MENDEL and GOLDSCHEIDER'S method was modified in the following details: Somogyis deproteinization was performed instead metaphosphoric acid as in the original method; to avoid the evaporation of the acetic aldehyde during the heating time with sulfuric acid a special glass stopped tube is proposed (Fig. 2). The reaction with sulfuric acid and veratrol is performed in an ice bath. Blood proteins precipitants were tried and Somogyi's lattest tecnic showed better results (Table V). Colorimetric readings were done in the PULFRICH photometer using filter S 53 and a 10 mm cup. The method is accurate within an error of 0.23 % and samples of 5 to 70 microg. could be easily determined. Normal values for human blood averaged 10.78 mg per 100 ml. More accurate results were obtained with the technic of MILLER & MUNTZ. Slight modifications were introduced: deproteinization with copper sulfate and sodium tungstate; satured p-hydroxydiphenyl solution according to KOENEMANN which is stable for 5 months when stored in the ice-box. Using the PULFRICH step-photometer the error is 0.17% with samples varying from 0.1 to 10 microg. of lactic acid. The filter employed was S 57 with the 5 mm cup. The method was adapted to 0.1 ml of blood. Normal values for human blood gave an average of 10.58 mg per 100 ml.

Plancto e hidrobiologia sanitária de tanques tropicais com dáfnias e rotíferos

The engineers of the modern University City are constructing a graceful bridge, named PONTE OSWALDO CRUZ, that crosses a portion of the Guanabara Bay (Fig. 1). The work at west pillar stopped for 3 years (The concret structure in Est. 1). As it will be seen from n.º 1 — 5 of the fig. 1, Est. I, the base of the structure will have five underground boxes of reinforcement, but, to-day they are just like as five uncovered water ponds, until at present: May 1963. (Est. I — fig. 3, n.º 3 — pond n.º 3; A. — old level of the water; B.— actual level of the water; c.— green water; E.— mass of bloom of blue algae Microcystis aeruginosa). Soon after SW portion, as 5 cells in series, of the pillar abutments, and also the NE portion nearly opposite in the Tibau Mount will be filled up with earth, a new way will link Rio City and the University City. We see to day Est. I, fig. 1 — the grasses on the half arenous beach of the Tibau Point. These natural Cyperaceae and Gramineae will be desappear because of so a new road, now under construction, when completed will be 33 feet above the mean sea level, as high as the pillar, covering exactly as that place. Although rainfall was the chief source of water for these ponds, the first water (before meterorological precipitations of whatever first rain it might fall) was a common tap water mixed with Portland Cement, which exuded gradually through the pores of the concret during its hardenning process. Some data of its first cement water composition are on the chemical table, and in Tab. n.º 4 and "Resultado n.º 1". The rain — receiving surface of each pond were about 15 by 16 feet, that is, 240 square feet; when they were full of water, their depth was of 2 feet 3", having each pond about 4,000 gallons. Climatic conditions are obviously similar of those of the Rio de Janeiro City: records of temperature, of precipitation and evaporation are seen on the graphics, figs. 2, 3, 4. Our conceptions of 4 phases is merely to satisfy an easy explanation thus the first phase that of exudation of concrete. We consider the 2nd. phase formation of bacterian and cyanophycean thin pellicel. 3rd. phase - dilution by rains, and fertilisation by birds; the 4th phase - plankton flora and fauna established. The biological material arrived with the air, the rains, and also with contaminations by dusts; with big portion of sand, of earth, and leaves of trees resulted of the SW wind actions in the storming days (See - Est. I, fig. 3, G. - the mangrove trees of the Pinheiro Island). Many birds set down and rest upon the pillar structure, its faeces which are good fertilizers fall into the ponds. Some birds were commonly pigeons, black ravens, swallows, sparrows and other sea mews, moor hens, and a few sea birds of comparatively rare occurence. We get only some examples of tropical dust contaminated helioplankton, of which incipient observations were been done sparcely. See the systematic list of the species of plankters. Phytoplankters - Cyanophyta algae as a basic part for food of zooplankters, represented chiefly by rotiferse, water-fleas Moinodaphnia and other Crustacea: Ostracoda Copepoda and Insecta: Chironomidae and Culicidae larvae. The polysaprobic of septic irruptions have not been done only by heating in summer, and, a good reason of that, for example: when the fifth pond was in polysaprobic phase as the same time an alike septic phase do not happened into the 3rd. pond, therefore, both were in the same conditions of temperature, but with unlike contaminations. Among the most important aquatic organisms used as indicatiors of pollution - and microorganisms of real importance in the field of sanitary science, by authorities of renown, for instance: PALMER, PRESCOTT, INGRAM, LIEBMANN, we choose following microalgae: a) The cosmopolite algae Scenedesmus quadricuada, a common indicator in mesosaprobio waters, which lives between pH 7,0 and it is assimilative of NO[3 subscripted] and NH[4 subscripted]. b) Species of the genus Chlamydomonas; it is even possible that all the species of theses genus inhabit strong-mesosaprobic to polysaprobic waters when in massive blooms. c) Several species of Euglenaceae in fast growing number, at the same time of the protozoa Amoebidae, Vorticellidae and simultaneous with deposition of the decaying cells of the blue algae Anacystis cyanea (= Microcystis) when the consumed oxygen by organic matter resulted in 40 mg. L. But, we found, among various Euglenacea the cosmopolite species (Euglena viridis, a well known polysaprobic indicatior of which presence occur in septic zone. d) Analcystis cyanea (= M. aeruginosa) as we observed was in blooms increasing to the order of billions of cells per litter, its maximum in the summer. Temperatures 73ºF to 82ºF but even 90ºF, the pH higher than 8. When these blue algae was joined to the rotifer Brachionus calyflorus the waters gets a milky appearance, but greenished one. In fact, that cosmopolite algae is used as a mesosaprobic indicator. Into the water of the ponds its predominance finished when the septic polysaprobic conditions began. e) Ankistrodesmus falcatus was present in the 5th pond from 26the. April untill the 26th July, and when N.NH[4 subscripted] gets 1.28 mg. L. and when chlorinity stayed from 0.034 to 0.061 mg. L. It never was found at N.NH[4 subscripted] higher than 1 mg. L. The green algae A. falcatus, an indicatior of pollution, lives in moderate mesosaprobic waters. f) As everyone knows, the rotifer eggs may be widely dispersed by wind. The rotifer Asplanchna brightwelli in our observation seemed like a green colored bag, overcharged by green cells and detritus, specially into its spacious stomach, which ends blindly (the intestine, cloaca, being absent). The stock of Asplanchna in the ponds, during the construction of the bridge "PONTE OSWALDO CRUZ" inhabits alkaline waters, pH 8,0 a 8,3, and when we observed we noted its dissolved oxygen from 3.5 to 4 mg. L. In these ponds Asplanchna lived in 0,2 P.PO[4 subscripted]. (Remember the hydobiological observations foreign to braslian waters refer only from 0.06 to 0,010 mg. L. P.PO[4 subscripted]; and they refer resistance to 0.8 N.NH[4 subscripted]). By our data, that rotiger resist commonly to 1.2 until 1.8 mg. L.N.NH[4 subscripted]; here in our ponds and, when NO[2 subscripted] appears Asplanchna desappears. It may be that Asplanchna were devoured by nitrite resistant animals of by Culicidae or other mosquitoes devoured by Due to these facts the number and the distribution of Asplanchna varies considerabley; see - plates of plankton successions. g) Brachionus one of the commonest members of class Rotatoria was frquently found in abundance into the ponds, and we notice an important biological change produce by the rotifer Brachonus colyciflorus: the occurence of its Brachionus clayciflorus forms pallas, is rare in Brazil, as we know about this. h) When we found the water flea MOinodaphnia we do not record simultanous presence of the blue algae Agmenellun (= Merismopedia).

Histogical and ultrastructural aspects of the brindley's glands of pantrongylus megistus (Burmeister, 1835) (Hemiptera: Reduviidae)

The Brindley's glands of Panstrongylus megistus were studied under the antomic, histologic and ultrastructural point of view. These glands located in the insect's methatorax are paired and have an opening near the third parir of the feet. Beside this aperture, ther are evaporation areas. Shape, sixe and aspect of the gland vary according to the feeding status. The glands are composed by a tubular part corresponding to the duct and a sack-like portion corrsponding to the secretory part. By electron microscopy we observed that the basal part of the epithelium has many interdigitations associated with mitochondria. On the apical surface where epicuticular foldings are located an electonlucent space is often seen. The glands are composed of the following elements: 1) superficial epithelial cells, located just below the apical surface foldings; 2) secretory cells; which are long and have an intracellular canalicule which changes according to the functional state of the cell; 3) a collecting duct to the secretory cells and covered with an epicuticle, reaching up to the gland's lumen; and 4) cells around the duct.

Current status of biotechnology in China

There has been increasing interest over past decade in exploring the possibility of using new biotechinology to produce new products and to improve the old productive process. The researches and applications of genetic engineering, cell fusion, mutagenesis, cell and enzyme immobilization in enzyme, antibiotic, vitamine, steroid, amino acid, organic acid, solvent, food and brewage industries is reviewed.

Evaluation of some organic compounds on bloodstream forms of Trypanosoma cruzi

Accidental transmission of Chagas' disease to man by blood transfusion is a serious problem in Latin-America. This paper describes the testing of several synthetic, semi-synthetic, and natural compounds for their activity against blood trypomastigotes in vitro at 4-C. The compounds embody several types of chemical structures: benzoquinone, naphthoquinone, anthracenequinone, phenanthrenequinone, imidazole, piperazine, quinoline, xanthene, and simple benzenic and naphthalenic derivates. Some of them are for the first time tested against Trypanosoma cruzi. The toxic effect these compounds on this parasite was done by two quite distinct sets of experiments. In one set, the compounds were added to infected blood as ethanolic solution. In this situation the most active one was a furan-1, 2-naphthoquinone, in the same range as gentian violet, a new fact to be considered in the assessment of structure-activity relationships in this class of compounds. In other set, we tentatively evaluated the biological activity of water insoluble compounds by adding them in a pure form without solvent into infected blood. In this way some appear to be very active and it was postulated that the effectiveness of such compounds must result from interactions between them and specific blood components.

Biodistribution Study of the Anaesthetic Sodium Phenobarbital Labelled with Technetium-99m in Swiss Mice Infected with Schistosoma mansoni Sambon, 1907

Technetium-99m (99mTc) is a radionuclide that has negligible enviromnental impact, is easily available, inexpensive and can be used as a radioactive tracer in biological experiences. In order to know the mode of action of sodium phenobarbital in moving adult Schistosoma mansoni worms from mesenteric veins to the liver, we labelled sodium phenobarbital (PBBT) with 99mTc and a biodistribution study in infected and non-infected Swiss mice was performed. The PBBT was incubated with stannous chloride used as reducing agent and with 99mTc, as sodium pertechnetate. The radioactivity labelling (%) was determined by paper ascending chromatography perfomed with acetone (solvent). The 99mTc-PBBT was administered by intraperitoneal route to Swiss mice infected eight weeks before. The animals were perfused after diferent periods of time (0,1,2,3,4 hr) when blood, spleen, liver, portal vein, mesenteric veins, stomach, kidneys and adult worms were isolated. The radioactivity present in these samples was counted in a well counter and the percentage was determined. The radioactivity was mainly taken up by the blood, kidney, liver and spleen. No radioactivity was found on the adult worms. We concluded that the worm shift was due to an action on the host of the sodium phenobarbital

Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods

The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20% of the AdV, 90% of the RV and 100% of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8% of the AdV and 90% of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

Utilização de bioensaios e marcadores moleculares para detecção da resistência de coleópteros de produtos armazenados a inseticidas

The qualitative and quantitative losses caused by stored product insects are of great concern, and since there is only a few active ingredients available for their control it is very important to have a frequent insect resistance monitoring. The objective of this research is to evaluate combination of bioassays and molecular marker techniques to detect insecticide resistance in stored product beetles. The Coleoptera species used for the tests were Sitophilus oryzae (L.) (Curculionidae), Rhyzopertha dominica (F.) (Bostrichidae) and Oryzaephilus surinamensis (L.) (Silvanidae). For the bioassays it was used the impregnated filter paper technique, applying 1 mL of deltamethrin (K-Obiol 25 CE TM) using four concentrations and five replicates, including a control with solvent only. Ten adults of each species were liberated separately on each dish. The mortality was evaluated after 24 h and resistance determined by probit analysis. The samples used for the PCR-RAPD were either in vivo or preserved in 70% ethanol, kept in -18°C freezer. After extraction, quantification and DNA quality analysis, the 25 µL samples had the DNA amplified and tested with six primers. The bioassays showed a crescent mortality proportional to insecticide concentration. The resistance factor for R. dominica, S. zeamais and S. oryzae were: 2,2; 3,2 and 9,2, respectively, compared to the susceptible populations of each species. The PCR-RAPD analysis revealed bands which indicate inter and intraspecific variability in the populations, but it was not possible to correlate them to resistance. The association of bioassay and PCR-RAPD represents a precise and valuable tool for resistance management of stored product insects, but more populations and primers should be tested.

Morphological study of the thorax of the five nymphal instars of Triatoma arthurneivai Lent & Martins (Hemiptera, Reduviidae, Triatominae)

Scanning electron microscope (SEM) was done in order to study dorsal, ventral and lateral sides of 1st, 2nd, 3rd, 4th and 5th nymphal instars of Triatoma arthurneivai. The five nymphal instar can be recognized based on different pronotum, mesonotum and metanotum shapes and characteristics. In the 1st instar collar, hairless areas and tubercles are absent. The 2 nd instar presents collar, hairless areas and tubercles. In the 3rd instar occurs the development of wing pads. In the 4 th instar the four wing pads are expanded, but do not reach the abdomen and in the 5th instar the anterior wing pads almost overlap the posterior ones. At the ventral side, two metasternal glands openings (1+1) were found in all five nymphal instars. Brindley's gland evaporation areas (1+1) are located at the mesopleuron, as well as an evaporation area is located at the propleuron in all nymphal instars (1+1).

Energy substrate used by workers of leaf-cutting ants during nest excavation

Energy substrate used by workers of leaf-cutting ants during nest excavation. In this study we aimed to ascertain whether leaf-cutting ant workers lose body reserves (fat or sugars) as a function of nest excavation. For each treatment, we isolated 10 workers of Atta sexdens into two experimental groups, Control (C- without excavation) and Soil (S- with excavation), which were kept for different time intervals (0, 24, 48 or 72 hours), totaling 700 tested workers. We then determined the concentration of soluble carbohydrates and total lipid content in them. The total carbohydrates were determined colorimetrically, based on the reaction between carbohydrates and sulfuric acid-phenol. For determination of lipids, the insects were immersed in organic solvent until they reached a constant weight. Our results showed that carbohydrates are consumed during nest excavation activities. In the experimental groups S24, S48 and S72, there was an average reduction of 5.82 (20.42%), 14.31 (44.96%) and 13.27 (43.96%) µ.mg-1 in soluble sugar when compared with the experimental groups that did not excavate. Furthermore, the lipids were not used during this activity. With respect to dry mass of the workers, their values were C0 = 8%, C24 = 10.4%, C48 = 9.2%, C72 = 10%, S24 = 9.2%, S48 = 8.7% and S72 = 8.5%. Our results show experimentally that the source of energy for nest excavation is carbohydrates, whereas lipids are conserved for other activities.