Chemical composition and analgesic activity of the leaves and branches of Marlierea tomentosa Camb.

This paper describes the isolation of five phytoconstituents from Marlierea tomentosa. The triterpenes α-amyrin (1) and β-amyrin (2), and the flavonoids quercitrin (3) and isoquercetin (4), were isolated from the leaves. The branches afforded the triterpene arjunolic acid (5). The antinociceptive activity of crude extract, fractions and compound 5 were tested using the writhing and formalin tests in mice. The crude extract, some fractions, particularly dichloromethane and butanol (leaves), ethyl acetate (branches) and arjunolic acid (5) (81.2% inhibition) were more active against the writhing test than the two reference drugs, acetylsalicylic acid and indomethacin.

Trypanocidal activity of Meliaceae and Rutaceae plant extracts

The in vitro trypanocidal activity of 22 extracts and 43 fractions of plants belonging to the families Meliaceae and Rutaceae was evaluated. The extracts from leaves of Conchocarphus heterophyllus and branches of Trichilia ramalhoi were the most active. The trypanocidal activity seems to be increased by fractionation of the extracts. Fractions from C. heterophyllus and Galipea carinata were the most active and a 100% lysis of the parasites was observed for five fractions. From one of them were isolated two flavonoids: flavone and 7-methoxyflavone, which showed weak trypanocidal activity. The results obtained from the extracts and fractions revealed that the order Rutales is a promising source for the search of new drugs for Chagas disease. Phytochemical studies with the other active fractions are underway in order to isolate compounds, which could be associated with observed activities.

Drimanes from Drimys brasiliensis with leishmanicidal and antimalarial activity

This paper evaluates CHCl3 and CH3OH extracts of the stem bark, branches and leaves of Drimys brasiliensis and drimane sesquiterpenes isolated from the stem bark against strains of Leishmania amazonensis and Leishmania braziliensis promastigotes and Plasmodium falciparum trophozoites. All of the extracts and compounds were tested in cell lines in comparison with reference standards and cell viability was determined by the XTT method. The CHCl3 and CH3OH extracts from the stem bark and branches yielded promising results against two strains of Leishmania, with 50% inhibitory concentrations (IC50 ) values ranging from 39-100 µg/mL. The CHCl3 extract of the stem bark returned IC50 values of 39 and 40.6 µg/mL for L. amazonensis and L. braziliensis, respectively. The drimanes were relatively effective: 1-β-(p-coumaroyloxy)-polygodial produced IC50 values of 5.55 and 2.52 µM for L. amazonensis and L. braziliensis, respectively, compared with 1-β-(p-methoxycinnamoyl)-polygodial, which produced respective IC50 values of 15.85 and 17.80 µM. The CHCl3 extract demonstrated activity (IC50 of 3.0 µg/mL) against P. falciparum. The IC50 values of 1-β-(p-cumaroyloxyl)-polygodial and 1-β-(p-methoxycinnamoyl)-polygodial were 1.01 and 4.87 µM, respectively, for the trophozoite strain. Therefore, the results suggest that D. brasiliensis is a promising plant from which to obtain new and effective antiparasitic agents.

Aspidosperma (Apocynaceae) plant cytotoxicity and activity towards malaria parasites. Part I: Aspidosperma nitidum (Benth) used as a remedy to treat fever and malaria in the Amazon

Infusions of Aspidosperma nitidum (Apocynaceae) wood bark are used to treat fever and malaria in the Amazon Region. Several species of this family are known to possess indole alkaloids and other classes of secondary metabolites, whereas terpenoids, an inositol and the indole alkaloids harmane-3 acid and braznitidumine have been described in A. nitidum . In the present study, extracts from the wood bark, leaves and branches of this species were prepared for assays against malaria parasites and cytotoxicity testing using human hepatoma and normal monkey kidney cells. The wood bark extracts were active against Plasmodium falciparum and showed a low cytotoxicity in vitro, whereas the leaf and branch extracts and the pure alkaloid braznitidumine were inactive. A crude methanol extract was subjected to acid-base fractionation aimed at obtaining alkaloid-rich fractions, which were active at low concentrations against P. falciparum and in mice infected with and sensitive Plasmodium berghei parasites. Our data validate the antimalarial usefulness of A. nitidum wood bark, a remedy that can most likely help to control malaria. However, the molecules responsible for this antimalarial activity have not yet been identified. Considering their high selectivity index, the alkaloid-rich fractions from the plant bark might be useful in the development of new antimalarials.

Triterpenes from Minquartia guianensis (Olacaceae) and in vitro antimalarial activity

Minquartia guianensis, popularly known as acariquara, was phytochemically investigated. The following triterpenes were isolated from the dichloromethane extract of leaves: lupen-3-one (1), taraxer-3-one (2) and oleanolic acid (3). The dichloromethane extract of branches yielded the triterpene 3β-methoxy-lup-20(29)-ene (4). The chemical structures were characterized by NMR data. Plant extracts, substance 3, squalene (5) and taraxerol (6), (5 and 6 previously isolated), were evaluated by in vitro assay against chloroquine resistant Plasmodium falciparum. The dichloromethane extract of leaves and the three triterpenes assayed have shown partial activity. Thus, these results demonstrated that new potential antimalarial natural products can be found even in partially active extracts.

Increased angiotensin-converting enzyme activity in the left ventricle after infarction

An increase in angiotensin-converting enzyme (ACE) activity has been observed in the heart after myocardial infarction (MI). Since most studies have been conducted in chronically infarcted individuals exhibiting variable degrees of heart failure, the present study was designed to determine ACE activity in an earlier phase of MI, before heart failure development. MI was produced in 3-month old male Wistar rats by ligation of the anterior branches of the left coronary artery, control rats underwent sham surgery and the animals were studied 7 or 15 days later. Hemodynamic data obtained for the anesthetized animals showed normal values of arterial blood pressure and of end-diastolic pressure in the right and left ventricular cavities of MI rats. Right and left ventricular (RV, LV) muscle and scar tissue homogenates were prepared to determine ACE activity in vitro by measuring the velocity of His-Leu release from the synthetic substrate Hyp-His-Leu. ACE activity was corrected to the tissue wet weight and is reported as nmol His-Leu g-1 min-1. No significant change in ACE activity in the RV homogenates was demonstrable. A small nonsignificant increase of ACE activity (11 &plusmn; 9%; P0.05) was observed 7 days after MI in the surviving left ventricular muscle. Two weeks after surgery, however, ACE activity was 46 &plusmn; 11% (P<0.05) higher in infarcted rats compared to sham-operated rats. The highest ACE activity was demonstrable in the scar tissue homogenate. In rats studied two weeks after surgery, ACE activity in the LV muscle increased from 105 &plusmn; 7 nmol His-Leu g-1 min-1 in control hearts to 153 &plusmn; 11 nmol His-Leu g-1 min-1 (P<0.05) in the remaining LV muscle of MI rats and to 1051 &plusmn; 208 nmol His-Leu g-1 min-1 (P<0.001) in the fibrous scar. These data indicate that ACE activity increased in the heart after infarction before heart failure was demonstrable by hemodynamic measurements. Since the blood vessels of the scar drain to the remaining LV myocardium, the high ACE activity present in the fibrous scar may increase the angiotensin II concentration and decrease bradykinin in the cardiac tissues surrounding the infarcted area. The increased angiotensin II in the fibrous scar may contribute to the reactive fibrosis and hypertrophy in the left ventricular muscle surviving infarction

Methylmercury intoxication and histochemical demonstration of NADPH-diaphorase activity in the striate cortex of adult cats

The effects of methylmercury (MeHg) on histochemical demonstration of the NADPH-diaphorase (NADPH-d) activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 µg MeHg and 100 g fish containing 0.03 µg MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 µg/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1) and the other animals were perfused 6 months later (group 2). After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80). Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg) in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P<0.05). These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter.

Exogenous induction of ovarian activity and ovulation and transfer of fresh embryos of domestic cat (Felis catus)

The objective of the present study was the exogenous stimulation of ovarian activity and definition of embryo collection, and transfer protocols, in the domestic cat for potential application in non-domestic endangered species. Sixteen adult queens and two adult male reproducers kept in the experimental cat house at the Morphology sector at the Veterinary Department (DVT), UFV, were used in this study. All the queens received a single application of 150 IU Equine Chorionic Gonadotropin (eCG) in the post estrus to induce ovarian activity and 80 to 84 hours later, received a single application of 100 UI Human Chorionic Gonadotropin (hCG) to induce ovulation. After hCG application, only the donor queens were naturally mated. The receptor queens received extra stimulus for induction of ovulation through manipulation of an intravaginal swab. Five to six days after hCG application, the donor queens were subjected to a laparotomy for embryo collection that was performed by trans-horn uterine washing. On average, six embryos were surgically inovulated. They were classified as type I and III compact morula and blastocysts in four receptor queens. Three animals presented pregnancy confirmed by ultrasound at day 36 and two of these animals gave birth to litters of two and four offsprings, respectively, at 66 and 63 days after induction of ovulation. Except for one still birth, all the offspring developed normally.

Antibacterial activity of Baccharis trimera (Less.) DC. (carqueja) against bacteria of medical interest

Baccharis trimera (Less.) (Asteraceae), popularly know as "carqueja", is a species commonly used in folk medicine for the treatment or prevention of diseases. In this context, the purpose of this work was to study the antibacterial activity of crude hydroalcoholic extract from Baccharis trimera against Gram-positive bacterial strains (Staphylococcus aureus ATCC 29213, Staphylococcus saprophyticus ATCC 15305, Staphylococcus epidermidis ATCC 12228, Enterococcus faecalis ATCC 19433) and Gram-negative bacteria (Escherichia coli EHEC ATCC 43895, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 27736, Salmonella typhi ATCC 19430) of clinical interest. Antibacterial susceptibility was evaluated by broth microdilution assay following the CLSI (formerly the NCCLS) guidelines. The extract from B. trimera showed antibacterial activity against Gram-positive bacteria and the most interesting result was obtained against S. epidermidis that presented Minimal Inhibitory Concentration of 250μg/mL. These results indicate that B. trimera have bacterisostatic potential against Gram-positive bacterial strains of medical interest and could serve as a base for further studies on the use of isolated compounds from this species as future antimicrobials.

Production and photosynthetic activity of Mimosa Verde and Mimosa Roxa lettuce in two farming systems

Lettuce (Lactuca sativa L.) is the most commonly consumed leaf vegetable in the Brazilian diet, and it is a good source of vitamins and minerals. It is widely grown in the conventional farming system. However, the hydroponic farming system has been gaining importance in the market, wining confidence from consumers, who are becoming increasingly more demanding on food quality. The objective of this study was to evaluate the performance of two lettuce cultivars on hydroponic and conventional farming systems for the production of fresh mass (FM) and dry mass (DM), photosynthesis, contents of chlorophyll and anthocyanin. The following two experiments were carried out: hydroponics farming (HF) and conventional farming (CF), performed in protect and unprotect environments, respectively, in Florianópolis, SC. Mimosa Verde cultivar (MV) showed greater fresh mass than Mimosa Roxa (MR), in both farming systems and the two cultivars presented better performance in the hydroponic system (287.7 g MV and 139.1 g MR) than the conventional system (129.7 g MV and 111.8 g MR). Mimosa Verde cultivar presented lower average contents of total chlorophyll (7.7 mg g-¹ FM) than Mimosa Roxa (11.8 mg g-¹FM), and both cultivars displayed higher means for this variable in the hydroponic farming system. Mimosa Roxa presented higher contents of anthocyanin in the conventional system (88.24 mg g-¹ FM) than the ones in the hydroponic system (36.89 mg g-¹ FM). The best results for CO2 net assimilation rate regarded to photosyntheticaly active photon flux density were found in the hydroponic system, for both lettuce cultivars. Variation in the contents of chlorophyll were also found. Those variations were higher in the protected system than in the hydroponic system and contents of anthocyanin were higher in the conventional system.

Application of GA3 and girdling of branches on the production of extemporaneous fruits of 'Tahiti' acid lime

The objective of this study was to evaluate the effect of girdling of branches and the application of GA3, still in the main crop flowering on the formation of reproductive structures, the fruit set of the following extemporaneous flowering of 'Tahiti' acid lime. This work used a randomized block design with five replications in a 4x3 factorial design. The first factor was the foliar application of GA3, 88 days before extemporaneous flowering at five concentrations (0, 7, 14 and 21 mg.L-1). The second factor was the girdling times, 108 days before flowering onset (A108AF), 78 days before flowering (A78AF) plus no-girdling control (NG). In extemporaneous flowering, the proportion of buds, flowers and fruits that abscised was 38.75, 31.25 and 18.64% respectively and 11.34% of all reproductive structures formed were collected. GA3 did not affect fruit set. Girdling increased fruit set by 229 and 256% at A108DAF and A78DAF respectively, in relation to NG plants. The GA3 reduced the formation of floral structures, which resulted in a quadratic increase in field production by reducing the competition for photoassimilates among fruits. The concentration of 21 mg.L-1 was the most productive, with 15.97 fruit.plant-1.

Biting activity of Aedes scapularis (Rondani) and Haemagogus mosquitoes in Southern Brazil (Diptera: Culicidae)

The biting activity of a population of Aedes scapularis (Rondani), Haemagogus capricornii Lutz and Hg. leucocelaenus (Dyar and Shannon) in Southern Brazil was studied between March 1980 and April 1983. Data were obtained with 25-hour human bait catches in three areas with patchy residual forests, named "Jacaré-Pepira", "Lupo" Farm, and "Sta. Helena" Farm, in the highland region of S. Paulo State (Brazil). Data obtained on Ae. scapularis were compared with those formerly gathered in the "Ribeira'' Valley lowlands, and were similar, except in the "Lupo" Farm study area, where a precrepuscular peak was observed, not recorded at the "Jacaré-Pepira" site or in the "Ribeira" Valley. In all the areas this mosquito showed diurnal and nocturnal activity, but was most active during the evening crepuscular period. These observations support the hypothesis about the successful adaptation of Ae. scapularis to man-made environments and have epidemiological implications that arise from it. As for Haemagogus, results obtained on the "Lupo" and "Sta. Helena" regions agree with previous data obtained in several other regions and show its diurnal activity. The proximity of "Lupo" Farm, where Hg. capricornii and Hg. leucocelaenus showed considerable activity, to "Araraquara" city where Aedes aegypti was recently found, raises some epidemiological considerations about the possibility of urban yellow fever resurgence.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 11 - Biting activity and blood-seeking parity of Anopheles (Kerteszia) in South-Eastern Brazil

An. (Ker.) cruzii and An. (Ker.) bellator were monitored by the use of human bait and Shannon trap collections during a one-year period in 1994. Indoor and outdoor collections were made on human bait and inside the forest environment a Shannon trap was used. Both were undertaken in the evening crepuscular period. Results showed a greater abundance of An. cruzii in the forest, where as An. bellator was more abundant in the domiciliary environment. Through the application of the Polovodova method an age grading was established. Computing the parous and nulliparous females with Christopher's Stage III and above, more than 30.0% of blood-seeking specimens of both anophelines had had a previous blood meal. The higher abundance of An. bellator as compared with that of An. cruzii in the domiciliary environment may be explained by the influence of the neighboring low-lying coastal islands of the estuary system.

Intra-population plasticity of Anopheles darlingi's (Diptera, Culicidae) biting activity patterns in the state of Amapá, Brazil

OBJECTIVE: To assess the variation in Anopheles darlingi's biting activity compared to An. marajoara in the same locality and to biting activity data from other regions. METHODS: Using human bait, eight observations of the biting activity of An. darlingi and An. marajoara were carried out during 1999 and 2000 in the municipality of São Raimundo do Pirativa, state of Amapá, Brazil. Each observation consisted of three consecutive 13-hour collections, close to full moon. There were shifts of collectors in the observation points and nocturnal periods. RESULTS: An. darlingi revealed considerable plasticity of biting activity in contrast to An. marajoara, which showed well-defined crepuscular biting peaks. No significant correlation between density and biting activity was found, but a significant correlation existed between time and proportional crepuscular activity, indicating underlying ecological processes not yet understood. Two of the four available data sets having multiple observations at one locality showed considerable plasticity of this species' biting patterns as well. CONCLUSION: Intra-population variation of biting activity can be as significant as inter-population variation. Some implications in malaria vector control and specific studies are also discussed.