... third. ABSTRACT Lopes, SA, Ribeiro, DM, Roberto, PG, França, SC, and Santos, JM 2000. ... sam... more ... third. ABSTRACT Lopes, SA, Ribeiro, DM, Roberto, PG, França, SC, and Santos, JM 2000. ... samples and the positive control. Popula-tion of viable X. fastidiosa cells varied from 4.9 × 104 to 6.6 × 105 CFU/g of to-bacco tissue. Phase ...
ABSTRACT Extracts of the medicinal plant Palicourea rigida Kunth, popularly known as douradinha, ... more ABSTRACT Extracts of the medicinal plant Palicourea rigida Kunth, popularly known as douradinha, are widely used for treating urinary tract disorders. Unfortunately, nowadays this is one of the species endemic to Brazilian Cerrado that is at greatest risk of extinction. The aim of the this work was to use AFLP molecular markers to determine the genetic structure and diversity of eight natural populations of P. rigida and to associate their genetic characteristics with loganin production in order to obtain provide relevant information to promote programs for the conservation of this valuable medicinal plant. A total of 120 polymorphic bands were scored and higher proportion of genetic diversity was found in inter-populations (64%) rather than in intra-populations (36%). Fst value was found to be significantly greater than zero (0.3601), demonstrating the complex genetic structure of P. rigida populations. Accessions collected from Cristalina, GO, showed higher percentage of polymorphic loci (65.5%) and the highest genetic diversity. Analysis of Molecular variance (AMOVA) demonstrated 63.9% of intra-population genetic variation. The lowest genetic variability was detected among accessions from the population found in Sacramento, MG. No spatial standard was observed for P. rigida population, suggesting a partially isolated island model. It was observed a minor but significant positive correlation (r = 0.22) between chemical and genetic matrices. The association between chemical and genetic data indicated that environmental factors promoted the loganin production in populations growing in Luziânia, GO, and therefore accessions from those populations should be considered as prime material for initiating the conservation process of P. rigida.
The HIV-tuberculosis co-infection has caused an impact on tuberculosis epidemiology all over the ... more The HIV-tuberculosis co-infection has caused an impact on tuberculosis epidemiology all over the world and the efficacies of the therapeutic schemes traditionally prescribed in the treatment of tuberculosis, such as isoniazid, rifampicin and pyrazinamide, have decreased due to the appearance of multidrug-resistant M. tuberculosis strains (MDR). This work is part of research on natural antimicrobial agents from plant extracts through bioassay-guided fractionation, by in vitro determination of the minimum inhibitory concentration (MIC) using the microdilution method with Alamar blue oxidation-reduction dye. Crude CHCl3Physalis angulata extracts and physalin-containing fractions displayed antimycobacterial activity against Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium malmoense and Mycobacterium intracellulare.
A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants h... more A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants has been established. The maximum number of shoots was obtained after 60 days of culture in Murashige & Skoog (MS) medium supplemented with 4.4 μM benzyladenine. Multiple rooting was achieved using MS medium with 2.4 μM 2-isopentyladenine. Wedelolactone was present in shoots cultured in media containing cytokinins.
ABSTRACT The quinone-methide triterpenoids (QMTs) are chemotaxonomic markers of the family Celast... more ABSTRACT The quinone-methide triterpenoids (QMTs) are chemotaxonomic markers of the family Celastraceae and many compounds of this class possess important anticancer and anti-inflammatory activities. However, the levels of QMTs in the roots of Celastraceous species are typically very low (ca. 0.0003 %) and commercial production by extraction from such tissues would not be commercially viable. With the aim of determining if cells cultured in vitro might provide alternative sources of these bioactive triterpenoids, we have quantified and compared the concentrations of QMTs accumulated by the roots of plants of Maytenus ilicifolia Mart. ex Reissek (Celastraceae) aged 0.5–10 years, and in callus and suspension cultures derived from a cell line that had been subcultured over a period of 10 years. Comparing plants cultivated in natura, the highest levels of QMTs were detected in the root bark of 5-year old specimens. However, cell suspensions derived from the long-term cell line retained their capacity to synthesize and accumulate QMTs, and presented levels of maytenin, 22β-hydroxymaytenin, celastrol and pristimerin that were, respectively, 1.96-, 2.48-, 8.85- and 3.29-times higher than those in the roots of 5-year old plants. The results presented herein open up new possibilities for the large-scale production of QMTs and for the development of novel pharmaceuticals.
Phospholipases A2 (PLA2) are multifunctional proteins which exhibit varied biological activities ... more Phospholipases A2 (PLA2) are multifunctional proteins which exhibit varied biological activities correlated to the structural diversities of the sub-classes. The crude aqueous extract from subterranean system of Mandevilla velutina, a plant found in Brazilian savanna, was assayed for its ability to inhibit biological activities of several snake venoms and isolated PLA2s. The extract induced total inhibition of the phospholipase activity of Crotalus durissus terrificus venom and only partial inhibition of Bothrops venoms. When assayed against purified toxins, the highest efficacy was detected against CB and crotoxin, while almost ineffective against PLA2s from the genus Bothrops. Although M. velutina crude extract significantly inhibited the myotoxic activity of C. d. terrificus venom and CB, it produced only partial inhibition of either Bothrops jararacussu venom or its main myotoxins BthTX-I (basic Lys49), BthTX-II (basic Asp49) and BthA-I-PLA2 (acidic Asp49). The extract exhibited also full inhibition of hemorrhage caused by Bothrops alternatus, Bothrops moojeni and Bothrops pirajai snake venoms, but partial inhibition (90%) of that induced by B. jararacussu venom. The extract was ineffective to inhibit the fibrinogenolytic activity of B. moojeni, B. alternatus and B. pirajai crude venoms, while their caseinolytic activity was only partially inhibited. No inhibition of the anticoagulant activity, although partial reduction of the edema-inducing activity of C. d. terrificus and B. alternatus crude venoms, CB, PrTX-I, BthTX-I and crotoxin was observed. Besides extending survival of mice injected with lethal doses of C. d. terrificus and B. jararacussu venoms, M. velutina extract decreased to 50% the lethality of mice. Extracts of 18 month old micropropagated plants were able to partially neutralize the effect of the crude venoms and toxins.
An algorithm for automatic clustering of database protein sequences from Bothrops jararacussu ven... more An algorithm for automatic clustering of database protein sequences from Bothrops jararacussu venomous gland, according to sequence similarities of their domains, is described. The program was written in C and Perl languages. This algorithm compares a domain with each ORF protein sequence in the database. Each nucleotide FASTA sequence generates six ORFs. As a result, the user has a list containing all sequences found in a specific domain and a display of the sequence, domain and number of hits. The algorithm lists only the sequences that present a minimum similarity of 30 hits and the best alignment. This limit was considered appropriate. The algorithm is available in the Internet (www.compbionet.org.br/cgi-domains/homesnake) and it can quickly and accurately organizes large database into classes.
Micropropagated shoots of Maytenus ilicifolia Mart. were obtained from axillary buds cultured in ... more Micropropagated shoots of Maytenus ilicifolia Mart. were obtained from axillary buds cultured in Murashige & Skoog medium supplemented with 13.3 μM 6-benzyladenine (BA). Addition of 1.1 μM 1-indole-3-acetic acid (IAA) to the medium increased shoot elongation. The number of shoots formed was influenced by BA concentration, degree of juvenility of the explant, and by bud explant position on the stem. Cultures of buds taken from stem parts located close to the shoot tip yielded more callus than shoots, whereas axillary buds at distant positions from the apical bud yielded more shoots.
Maytenus aquifolium (Celastraceae) and Salacia campestris (Hippocrateaceae) species accumulate fr... more Maytenus aquifolium (Celastraceae) and Salacia campestris (Hippocrateaceae) species accumulate friedelane and quinonemethide triterpenoids in their leaves and root bark, respectively. Enzymatic extracts obtained from leaves displayed cyclase activity with conversion of the substrate oxidosqualene to the triterpenes, 3β-friedelanol and friedelin. In addition, administration of (±)5-3H mevalonolactone in leaves of M. aquifolium seedlings produced radio labelled friedelin in the leaves, twigs and stems, while the root bark accumulated labelled maytenin and pristimerin. These experiments indicated that the triterpenes once biosynthesized in the leaves are translocated to the root bark and further transformed to the antitumoral quinonemethide triterpenoids.
The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using ... more The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium supplemented with 0.5 mg l-1 6-benzyladenine, 0.1 mg l-1 gibberelic acid added with 10 g l-1 sucrose. Rooting was achieved using MS medium devoid of growth regulators. An anatomical study confirmed shoot regeneration via direct organogenesis.
In order to produce explants of Mandevilla illustris (Vell) Woodson for the “Cerrado in vitro”, t... more In order to produce explants of Mandevilla illustris (Vell) Woodson for the “Cerrado in vitro”, the Germplasm Bank of UNAERP, we carried out a micropropagation protocol using MS or MS/3 medium supplemented with different concentrations of 6-benzyladeninepurine (BA), Zeatin or 2-isopentenyladenine for nodal segment growth, and α-naphthaleneacetic acid, indole-3-butyric acid (IBA) or 1,4 dithiothreitol for rooting. For nodal segments, all the cytokinins tested yielded similar results. However, 2.22 µM BA is more economical to use. MS/3 medium supplemented with 0.49 µM IBA was the most appropriate medium for rooting, resulting in 29% rooted explants. The crude aqueous extract from the subterranean system (SS) of M. illustris was assayed for its inhibitory action on the enzymatic activity of Crotalus durissus terrificus snake venom, isolated basic phospholipase A2 (CB) and crotoxin. It totally inhibited the phospholipase activity of crude Cdt venom and CB toxin and inhibited the phospholipase activity of crotoxin by 49%. The toxic action of both the crude venom and crotoxin was partially inhibited—there was a prolonged survival time and a 40.0% decrease in lethality.
... third. ABSTRACT Lopes, SA, Ribeiro, DM, Roberto, PG, França, SC, and Santos, JM 2000. ... sam... more ... third. ABSTRACT Lopes, SA, Ribeiro, DM, Roberto, PG, França, SC, and Santos, JM 2000. ... samples and the positive control. Popula-tion of viable X. fastidiosa cells varied from 4.9 × 104 to 6.6 × 105 CFU/g of to-bacco tissue. Phase ...
ABSTRACT Extracts of the medicinal plant Palicourea rigida Kunth, popularly known as douradinha, ... more ABSTRACT Extracts of the medicinal plant Palicourea rigida Kunth, popularly known as douradinha, are widely used for treating urinary tract disorders. Unfortunately, nowadays this is one of the species endemic to Brazilian Cerrado that is at greatest risk of extinction. The aim of the this work was to use AFLP molecular markers to determine the genetic structure and diversity of eight natural populations of P. rigida and to associate their genetic characteristics with loganin production in order to obtain provide relevant information to promote programs for the conservation of this valuable medicinal plant. A total of 120 polymorphic bands were scored and higher proportion of genetic diversity was found in inter-populations (64%) rather than in intra-populations (36%). Fst value was found to be significantly greater than zero (0.3601), demonstrating the complex genetic structure of P. rigida populations. Accessions collected from Cristalina, GO, showed higher percentage of polymorphic loci (65.5%) and the highest genetic diversity. Analysis of Molecular variance (AMOVA) demonstrated 63.9% of intra-population genetic variation. The lowest genetic variability was detected among accessions from the population found in Sacramento, MG. No spatial standard was observed for P. rigida population, suggesting a partially isolated island model. It was observed a minor but significant positive correlation (r = 0.22) between chemical and genetic matrices. The association between chemical and genetic data indicated that environmental factors promoted the loganin production in populations growing in Luziânia, GO, and therefore accessions from those populations should be considered as prime material for initiating the conservation process of P. rigida.
The HIV-tuberculosis co-infection has caused an impact on tuberculosis epidemiology all over the ... more The HIV-tuberculosis co-infection has caused an impact on tuberculosis epidemiology all over the world and the efficacies of the therapeutic schemes traditionally prescribed in the treatment of tuberculosis, such as isoniazid, rifampicin and pyrazinamide, have decreased due to the appearance of multidrug-resistant M. tuberculosis strains (MDR). This work is part of research on natural antimicrobial agents from plant extracts through bioassay-guided fractionation, by in vitro determination of the minimum inhibitory concentration (MIC) using the microdilution method with Alamar blue oxidation-reduction dye. Crude CHCl3Physalis angulata extracts and physalin-containing fractions displayed antimycobacterial activity against Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium malmoense and Mycobacterium intracellulare.
A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants h... more A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants has been established. The maximum number of shoots was obtained after 60 days of culture in Murashige & Skoog (MS) medium supplemented with 4.4 μM benzyladenine. Multiple rooting was achieved using MS medium with 2.4 μM 2-isopentyladenine. Wedelolactone was present in shoots cultured in media containing cytokinins.
ABSTRACT The quinone-methide triterpenoids (QMTs) are chemotaxonomic markers of the family Celast... more ABSTRACT The quinone-methide triterpenoids (QMTs) are chemotaxonomic markers of the family Celastraceae and many compounds of this class possess important anticancer and anti-inflammatory activities. However, the levels of QMTs in the roots of Celastraceous species are typically very low (ca. 0.0003 %) and commercial production by extraction from such tissues would not be commercially viable. With the aim of determining if cells cultured in vitro might provide alternative sources of these bioactive triterpenoids, we have quantified and compared the concentrations of QMTs accumulated by the roots of plants of Maytenus ilicifolia Mart. ex Reissek (Celastraceae) aged 0.5–10 years, and in callus and suspension cultures derived from a cell line that had been subcultured over a period of 10 years. Comparing plants cultivated in natura, the highest levels of QMTs were detected in the root bark of 5-year old specimens. However, cell suspensions derived from the long-term cell line retained their capacity to synthesize and accumulate QMTs, and presented levels of maytenin, 22β-hydroxymaytenin, celastrol and pristimerin that were, respectively, 1.96-, 2.48-, 8.85- and 3.29-times higher than those in the roots of 5-year old plants. The results presented herein open up new possibilities for the large-scale production of QMTs and for the development of novel pharmaceuticals.
Phospholipases A2 (PLA2) are multifunctional proteins which exhibit varied biological activities ... more Phospholipases A2 (PLA2) are multifunctional proteins which exhibit varied biological activities correlated to the structural diversities of the sub-classes. The crude aqueous extract from subterranean system of Mandevilla velutina, a plant found in Brazilian savanna, was assayed for its ability to inhibit biological activities of several snake venoms and isolated PLA2s. The extract induced total inhibition of the phospholipase activity of Crotalus durissus terrificus venom and only partial inhibition of Bothrops venoms. When assayed against purified toxins, the highest efficacy was detected against CB and crotoxin, while almost ineffective against PLA2s from the genus Bothrops. Although M. velutina crude extract significantly inhibited the myotoxic activity of C. d. terrificus venom and CB, it produced only partial inhibition of either Bothrops jararacussu venom or its main myotoxins BthTX-I (basic Lys49), BthTX-II (basic Asp49) and BthA-I-PLA2 (acidic Asp49). The extract exhibited also full inhibition of hemorrhage caused by Bothrops alternatus, Bothrops moojeni and Bothrops pirajai snake venoms, but partial inhibition (90%) of that induced by B. jararacussu venom. The extract was ineffective to inhibit the fibrinogenolytic activity of B. moojeni, B. alternatus and B. pirajai crude venoms, while their caseinolytic activity was only partially inhibited. No inhibition of the anticoagulant activity, although partial reduction of the edema-inducing activity of C. d. terrificus and B. alternatus crude venoms, CB, PrTX-I, BthTX-I and crotoxin was observed. Besides extending survival of mice injected with lethal doses of C. d. terrificus and B. jararacussu venoms, M. velutina extract decreased to 50% the lethality of mice. Extracts of 18 month old micropropagated plants were able to partially neutralize the effect of the crude venoms and toxins.
An algorithm for automatic clustering of database protein sequences from Bothrops jararacussu ven... more An algorithm for automatic clustering of database protein sequences from Bothrops jararacussu venomous gland, according to sequence similarities of their domains, is described. The program was written in C and Perl languages. This algorithm compares a domain with each ORF protein sequence in the database. Each nucleotide FASTA sequence generates six ORFs. As a result, the user has a list containing all sequences found in a specific domain and a display of the sequence, domain and number of hits. The algorithm lists only the sequences that present a minimum similarity of 30 hits and the best alignment. This limit was considered appropriate. The algorithm is available in the Internet (www.compbionet.org.br/cgi-domains/homesnake) and it can quickly and accurately organizes large database into classes.
Micropropagated shoots of Maytenus ilicifolia Mart. were obtained from axillary buds cultured in ... more Micropropagated shoots of Maytenus ilicifolia Mart. were obtained from axillary buds cultured in Murashige & Skoog medium supplemented with 13.3 μM 6-benzyladenine (BA). Addition of 1.1 μM 1-indole-3-acetic acid (IAA) to the medium increased shoot elongation. The number of shoots formed was influenced by BA concentration, degree of juvenility of the explant, and by bud explant position on the stem. Cultures of buds taken from stem parts located close to the shoot tip yielded more callus than shoots, whereas axillary buds at distant positions from the apical bud yielded more shoots.
Maytenus aquifolium (Celastraceae) and Salacia campestris (Hippocrateaceae) species accumulate fr... more Maytenus aquifolium (Celastraceae) and Salacia campestris (Hippocrateaceae) species accumulate friedelane and quinonemethide triterpenoids in their leaves and root bark, respectively. Enzymatic extracts obtained from leaves displayed cyclase activity with conversion of the substrate oxidosqualene to the triterpenes, 3β-friedelanol and friedelin. In addition, administration of (±)5-3H mevalonolactone in leaves of M. aquifolium seedlings produced radio labelled friedelin in the leaves, twigs and stems, while the root bark accumulated labelled maytenin and pristimerin. These experiments indicated that the triterpenes once biosynthesized in the leaves are translocated to the root bark and further transformed to the antitumoral quinonemethide triterpenoids.
The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using ... more The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium supplemented with 0.5 mg l-1 6-benzyladenine, 0.1 mg l-1 gibberelic acid added with 10 g l-1 sucrose. Rooting was achieved using MS medium devoid of growth regulators. An anatomical study confirmed shoot regeneration via direct organogenesis.
In order to produce explants of Mandevilla illustris (Vell) Woodson for the “Cerrado in vitro”, t... more In order to produce explants of Mandevilla illustris (Vell) Woodson for the “Cerrado in vitro”, the Germplasm Bank of UNAERP, we carried out a micropropagation protocol using MS or MS/3 medium supplemented with different concentrations of 6-benzyladeninepurine (BA), Zeatin or 2-isopentenyladenine for nodal segment growth, and α-naphthaleneacetic acid, indole-3-butyric acid (IBA) or 1,4 dithiothreitol for rooting. For nodal segments, all the cytokinins tested yielded similar results. However, 2.22 µM BA is more economical to use. MS/3 medium supplemented with 0.49 µM IBA was the most appropriate medium for rooting, resulting in 29% rooted explants. The crude aqueous extract from the subterranean system (SS) of M. illustris was assayed for its inhibitory action on the enzymatic activity of Crotalus durissus terrificus snake venom, isolated basic phospholipase A2 (CB) and crotoxin. It totally inhibited the phospholipase activity of crude Cdt venom and CB toxin and inhibited the phospholipase activity of crotoxin by 49%. The toxic action of both the crude venom and crotoxin was partially inhibited—there was a prolonged survival time and a 40.0% decrease in lethality.
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