A new benzophenone, and the antiplasmodial activities of the constituents of Securidaca longipedunculata fresen (Polygalaceae).

Extracts from Securidaca longipedunculata showed antiplasmodial activities against reference clones and clinical isolates using SYBR Green I method. A new benzophenone, 2,3,4,5-tetramethoxybenzophenone (1) was isolated and characterized along with seven known compounds: 4-hydroxy-2,3-dimethoxybenzophenone (2); 3-hydroxy-5-methoxybiphenyl (3), methyl-2-hydroxy-6-methoxybenzoate (4), benzyl-2-hydroxy-6-methoxybenzoate (5), 2-hydroxy-6-methoxybenzoic acid (6), 2,4,5-trimethoxybenzophenone (7) and 2-methoxy-3,4-methylenedioxybenzophenone (8). Compounds 1 and 2 showed ex vivo antiplasmodial activities (IC50 28.8 µM and 18.6 µM, respectively); while 5 and 8 showed in vivo activities (IC50 19.7 µM and 14.5 µM, respectively) against D6 strain. In a cytotoxicity assay, all the extracts (with an exception of the MeOH extract of the leaves) and pure compounds were not toxic to the normal LO2 and BEAS cell-lines, while the methanol roots extract (IC50 66.4 µg/mL against A549, and 77.4 µg/mL against HepG2), compounds 6 (IC50 22.2 µM against A549) and 7 (IC50 45.2 µM against HepG2) were weakly active against cancerous cell-lines.

Mycoheterotrophic Epirixanthes (Polygalaceae) has a typical angiosperm mitogenome but unorthodox plastid genomes.

BACKGROUND AND AIMS: Fully mycoheterotrophic plants derive carbon and other nutrients from root-associated fungi and have lost the ability to photosynthesize. While mycoheterotroph plastomes are often degraded compared with green plants, the effect of this unusual symbiosis on mitochondrial genome evolution is unknown. By providing the first complete organelle genome data from Polygalaceae, one of only three eudicot families that developed mycoheterotrophy, we explore how both organellar genomes evolved after loss of photosynthesis. METHODS: We sequenced and assembled four complete plastid genomes and a mitochondrial genome from species of Polygalaceae, focusing on non-photosynthetic Epirixanthes. We compared these genomes with those of other mycoheterotroph and parasitic plant lineages, and assessed whether organelle genes in Epirixanthes experienced relaxed or intensified selection compared with autotrophic relatives. KEY RESULTS: Plastomes of two species of Epirixanthes have become substantially degraded compared with that of autotrophic Polygala. Although the lack of photosynthesis is presumably homologous in the genus, the surveyed Epirixanthes species have marked differences in terms of plastome size, structural rearrangements, gene content and substitution rates. Remarkably, both apparently replaced a canonical plastid inverted repeat with large directly repeated sequences. The mitogenome of E. elongata incorporated a considerable number of fossilized plastid genes, by intracellular transfer from an ancestor with a less degraded plastome. Both plastid and mitochondrial genes in E. elongata have increased substitution rates, but the plastid genes of E. pallida do not. Despite this, both species have similar selection patterns operating on plastid housekeeping genes. CONCLUSIONS: Plastome evolution largely fits with patterns of gene degradation seen in other heterotrophic plants, but includes highly unusual directly duplicated regions. The causes of rate elevation in the sequenced Epirixanthes mitogenome and of rate differences in plastomes of related mycoheterotrophic species are not currently understood.

Anti-ophidian activity of Bredemeyera floribunda Willd. (Polygalaceae) root extract on the local effects induced by Bothrops jararacussu venom

Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3′,4′-OH flavonoids are known inhibitors of phospholipases A2.

Anti-ophidian activity of Bredemeyera floribunda Willd. (Polygalaceae) root extract on the local effects induced by Bothrops jararacussu venom.

Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3',4'-OH flavonoids are known inhibitors of phospholipases A2.

Susceptibility of Selected Multi-Drug Resistant Clinical Isolates to Leaves of Carpolobia lutea.

BACKGROUND: The decline in the production of new effective antibiotics coupled with the constantly evolving antimicrobial resistance remains a public health concern. This study thus evaluated the antibacterial and antifungal effects of the ethanolic, n-hexane and hot aqueous extracts of Carpolobialutea leaves. METHODS: The extracts were tested using agar well diffusion method against selected clinical isolates: Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, Staphylococcus aureus and Candida albicans. Antibiogram profile of the isolates were deduced by disc diffusion method. RESULTS: Multi-drug resistance was confirmed in all bacteria with a notable pandrug resistance in Pseudomonas aeruginosa. Ofloxacin, Erythromycin and Gentamicin were effective on two or three organisms, notably on Salmonella typhi and Escherichia coli. The preliminary antibacterial assay marked the efficacy of the ethanol and n-hexane extracts except on E. coli, with zero activity for hot water extracts at the stock concentration (200 mg/ml). Pseudomonas aeruginosaand Candida albicans were susceptible to lesser concentrations of the ethanol extracts at 5 mg/ml and 25 mg/ml respectively. None of the isolates showed sensitivity to lesser concentrations of n-hexane extract. Carpolobia lutea leaves proved to be effective over the use of antibiotics in inhibiting the activity of Pseudomonas aeruginosa which was resistant to the latter. The Minimum Inhibitory concentration of the ethanolic extract was considerably low (≤5mg/ml for P. aeruginosa and 25mg/ml for C. albicans). However, there was no Minimum Bactericidal concentration for the extracts against the clinical isolates. CONCLUSION: Carpolobia lutea shelters bioactive components with pharmacological potentials that could show efficiency in the treatment of bacterial infections.

Asemeia ovata (Polygalaceae): Quantitative determination and evaluation in silico of identified substances by HPLC-DAD.

BACKGROUND: In Brazil, the Asemeia genus has 19 species (12 endemic) and 2 varieties (both endemic) and some of them are found in semi-arid Bahia. OBJECTIVE: The objective of this study was to quantitatively determine identified substances by HPLC-DAD in Asemeia ovata extracts and to predict their biological activities in silico. METHOD: The quantification method by HPLC-DAD has been validated according to the guidelines of the International Conference of Harmonization. The prediction in silico activities was made by Target Fishing methods (TF), followed by docking by the program DOCK 6.7 and assessment of interaction profiles for Protein-Ligand Interaction Profiler server. RESULTS: It was possible to identify and quantify using HPLC-DAD substances: rutin, luteolin-7-O-glucoside, caffeic acid, p-coumaric acid and trans-ferulic acid. The ChemProt 2.0 server was selected for TF method, which has shown potential activity of compounds on molecular targets such as Carbonic anhydrase 12, epidermal growth factor receptor and sodium-glucose cotransporter 2. CONCLUSION: This work provides new results for the species both from a biological and chemical point of view, and has interesting potential to be discovered with the prospect of further studies.

Anti-acetylcholinesterase activity and antioxidant properties of extracts and fractions of Carpolobia lutea.

CONTEXT: There is an unmet need to discover new treatments for Alzheimer's disease. This study determined the anti-acetylcholinesterase (AChE) activity, DPPH free radical scavenging and antioxidant properties of Carpolobia lutea G. Don (Polygalaceae). OBJECTIVE: The objective of this study is to quantify C. lutea anti-AChE, DPPH free radical scavenging, and antioxidant activities and cell cytotoxicity. MATERIALS AND METHODS: Plant stem, leaves and roots were subjected to sequential solvent extractions, and screened for anti-AChE activity across a concentration range of 0.02-200 µg/mL. Plant DPPH radical scavenging activity, reducing power, and total phenolic and flavonoid contents were determined, and cytotoxicity evaluated using human hepatocytes. RESULTS: Carpolobia lutea exhibited concentration-dependent anti-AChE activity. The most potent inhibitory activity for the stem was the crude ethanol extract and hexane stem fraction oil (IC50 = 140 µg/mL); for the leaves, the chloroform leaf fraction (IC50 = 60 µg/mL); and for roots, the methanol, ethyl acetate and aqueous root fractions (IC50 = 0.3-3 µg/mL). Dose-dependent free radical scavenging activity and reducing power were observed with increasing stem, leaf or root concentration. Total phenolic contents were the highest in the stem: ∼632 mg gallic acid equivalents/g for a hexane stem fraction oil. Total flavonoid content was the highest in the leaves: ∼297 mg quercetin equivalents/g for a chloroform leaf fraction. At 1 µg/mL, only the crude ethanol extract oil was significantly cytotoxic to hepatocytes. DISCUSSION AND CONCLUSIONS: Carpolobia lutea possesses anti-AChE activity and beneficial antioxidant capacity indicative of its potential development as a treatment of Alzheimer's and other diseases characterized by a cholinergic deficit.

Carpolobia lutea G. Don (Polygalaceae) Inhibits Inflammation and Oxidative Stress in an Acetic Acid Induced Model of Rat Colitis.

Carpolobia lutea G. Don (CL) is used in folk medicine for the treatment of inflammatory disorders. Effort in this study is to evaluate the beneficial effect of the aqueous-methanol extract of leaf of CL in a rat model of acetic acid induced colitis. Male Wistar rats were distributed into 6 groups of 7 rats each; non colitic, untreated colitic and colitic rats treated with graded doses of CL (100-800 mg/kg). Rats were pre-treated for 2 days before colitis induction and thereafter for 7 days post colitic induction. 24 h after the last treatment, animals were sacrificed and colonic inflammation was evaluated both macroscopically and biochemically. Macroscopic damage score, weight/length ratio, myeloperoxidase (MPO) activity and tumor necrotic factor alpha (TNF-α) levels, were significantly higher in untreated colitic rats in comparison with non colitic rats (P<0.05). Treatment with CL significantly reduced the macroscopic damage scores, neutrophil infiltration (MPO activity) and TNF-α level (P<0.05). In addition, C. lutea significantly prevented depletion of colonic GSH and (SOD) levels (P<0.05). It appears that the beneficial effect of methanol extract of C. lutea leaf observed in this study is dose dependent and is related to its antioxidant and anti-inflammatory activity.

Effects of aqueous root extract of Carpolobia alba G. Don on sexual behaviour in adult male rats.

Several lines say that a number of natural products, mostly plant based, have been claimed to cure male sexual dysfunction. One of such botanicals is Carpolobia alba G. Don which is used in Cameroon to treat erectile dysfunction and related male sexual debilities. However, the traditional claim has not been scientifically tested. Thirty adult male rats (five groups of six animals) were orally treated daily with distilled water, sildenafil citrate (positive control), 75, 150, 300 mg/kg of C. alba G. Don roots aqueous extract. The erection and ejaculation properties were recorded on the first, seventh and fourteenth day of treatment by following copulation settings: number of erections (NE), mount frequency (MF), intromission frequency (IF), ejaculation frequency (EF), mount latency (ML), intromission latency (IL), ejaculation latency (EL), average interval of copulation (AIC) and post-ejaculatory interval (PEI). The extract resulted in an improvement of copulation parameters through significant reduction (p < .001) of PEI, ML and IL and significant increase (p < .001) in the NE, MF, IF, EF, EL and AIC. These results indicate a pro-erectile and pro-ejaculatory potential of aqueous root extract of this plant in male rats.

Morphology, ontogeny and structure of the stipular nectaries in Caamembeca spectabilis (Polygalaceae) / Morfologia, ontogenia e estrutura dos nectários estipulares em Caamembeca spectabilis (Polygalaceae)

Nodal glands are found in one third of the Polygalaceae genera and have valuable taxonomic, ecological and evolutionary significance. In Brazil, they occur in five of the eleven genera already registered. However, there is still a controversy regarding the origin of these structures. The objective of this study was to characterize the morphology and the origin of nodal glands inCaamembeca spectabilis, in order to increase the structural and functional knowledge of these glands in the genera. Samples of nodal regions were collected, fixed and processed according to the methods of light microscopy and electron scanning. Ants were observed and identified along the stem axis. The glucose in exudate allows us to classify these glands as extrafloral nectaries. They are located in pairs on the nodal region. However, its origin is in the leaf trace. In the longitudinal section, the nectaries were present in the apex of cells with anticlinal walls impregnated with suberin, which represents the first record for the family. In this region there is also the formation of a hole by lysis. The secretory tissue is surrounded by phloem. Xylem vessels were observed only on the basis of the nectary, where there are also idioblasts with crystals in druse type. We have studied the ontogeny of the glands nodal in Caamembeca spectabilis and unveiled that these glands are linked to the leaves as stipular nectaries. In addition, the new findings presented here may add support for the understanding of morphology and anatomy of nodal glands in Caamembeca.

Glândulas nodais são encontradas em um terço dos gêneros de Polygalaceae e possuem grande importância taxonômica, ecológica e evolutiva. No Brasil, ocorrem em cinco, dos onze gêneros já registrados. Contudo, ainda há controvérsias quanto à origem dessas estruturas. O objetivo deste trabalho foi caracterizar a morfologia e a origem das glândulas nodais em Caamembeca spectabilis, visando ampliar o conhecimento estrutural e funcional dessas glândulas no gênero. Amostras das regiões nodais foram coletadas, fixadas e processadas segundo os métodos em microscopia de luz e eletrônica de varredura. Formigas foram observadas e identificadas ao longo do eixo caulinar. A glicose no exsudato permite tratar as glândulas como nectários extraflorais. Esses estão localizados aos pares na região nodal. Porém, sua origem ocorre no traço foliar. Os nectários, em secção longitudinal, apresentam as células do ápice com paredes anticlinais impregnadas com suberina, sendo este o primeiro registro para a família. Nessa região também ocorre a formação de um orifício por lise. O tecido secretor do nectário é circundado pelo floema. Vasos de xilema foram observados apenas na base do nectário, onde também ocorrem idioblastos com cristais do tipo drusas. Dessa forma, este estudo confirma a trajetória ontogenética das glândulas nodais em C. spectabilis, as quais, de fato, estão ligadas às folhas, como nectários estipulares. Além de apresentar dados inéditos que auxiliam a compreensão morfológica e anatômica das mesmas em Caamembeca.

Atividade alelopática dos lixiviados de Asemeia extraaxillaris (Polygalaceae) sobre o crescimento de Ipomoea cordifolia / Allelopathic activity of leachates from Asemeia extraaxillaris (Polygalaceae) on the growth of Ipomoea cordifolia

RESUMO O objetivo desse trabalho foi avaliar a atividade alelopática dos lixiviados das raízes e folhas secas de Asemeia extraaxillaris sobre o crescimento de corda de viola Ipomoea cordifolia em casa de vegetação, para avaliação dos efeitos sobre o estresse oxidativo. Nos bioensaios foram utilizadas três concentrações (0,25 - 5,00 e 10,00 g) e um controle, com quatro repetições de 10 plantas, cultivadas em solo durante 30 dias. Os lixiviados possuem ação alelopática e o maior efeito foi verificado para o pó das raizes. O efeito inibitório foi dose-dependente e o aumento do estresse oxidativo foi observado pelos níveis da catalase, peroxidase, superóxido dismutase, glutationa redutase, peroxidação lipídica e síntese de formazan. Concluiu-se que A. extraaxillaris possui substâncias capazes de afetar a emergência, o crescimento e o estresse oxidativo de I. cordifolia.

ABSTRACT The purpose of this article is to assess the allelopathic activity of leachates from the roots and dried leaves of Asemeia extraaxillaris on the germination and growth of corda de viola (Ipomoea cordifolia) in a greenhouse, and evaluate its effect on oxidative stress. For the bioassays, three concentrations (0.25, 5.00, and 10.00 g) and one control were used, with four replications of 10 plants, cultivated in soil for 30 days. The leachates exhibited allelopathic action, and the greatest effect was found in the root powder. The inhibitory effect was dose-dependent and the increase in oxidative stress was observed by the levels of catalase, peroxidase, superoxide dismutase, glutathione reductase, lipid peroxidation, and formazan synthesis. It was affirmed that A. extraaxillaris contains substances capable of affecting seedling emergence, growth, and oxidative stress in I. cordifolia.

Evolution of mycoheterotrophy in Polygalaceae: The case of Epirixanthes.

PREMISE OF THE STUDY: The mycoheterotrophic lifestyle has enabled some plant lineages to obtain carbon from their mycorrhizal symbionts. The mycoheterotrophic genus Epirixanthes (Polygalaceae) consists of six species from tropical Asia. Although it is probably closely related to the chlorophyllous genus Salomonia and linked to arbuscular mycorrhizal fungi, lack of DNA sequence data has thus far prevented these hypotheses from being tested. Therefore, the evolutionary history of Epirixanthes remains largely unknown. METHODS: We reconstructed the phylogenetic relationships of Epirixanthes based on nuclear ITS and plastid matK data. Divergence times were inferred using a Bayesian relaxed clock approach, and we phylogenetically analyzed its mycorrhizal symbionts. We furthermore assigned these symbionts to operational taxonomic units, compared them with symbionts of other Polygalaceae, and measured their phylogenetic diversity. KEY RESULTS: We found that Epirixanthes is placed in tribe Polygaleae as sister to Salomonia. Epirixanthes has a Miocene-Oligocene stem age and grows exclusively in symbiosis with fungi of Glomeraceae. Salomonia and some Polygala species are linked to both Glomeraceae and Acaulosporaceae, resulting in higher phylogenetic diversity values. The majority of the symbionts of Epirixanthes are not found in Salomonia or Polygala, although a few shared fungal taxa are found. CONCLUSIONS: Epirixanthes forms a relatively young mycoheterotrophic lineage. The Oligocene-Miocene origin suggests its evolution was influenced by the environmental dynamics in Southeast Asia during this time. Although comparison of fungi from Epirixanthes with those from Salomonia and Polygala suggests some specialization, many other mycoheterotrophic plants are linked to a more narrow set of Glomeraceae.

Xanthones from the roots of Moutabea guianensis Aubl.

The phytochemical investigation of Moutabea guianensis roots led to the isolation of five polyoxygenated xanthones, including two new ones named moutabeone B (1,8-dihydroxy-4,5,6,7-tetramethoxyxanthone) and moutabeone C (1-hydroxy-4,5,6,7,8-pentamethoxyxanthone), along with the three known xanthones, 1,8-dihydroxy-4,6-dimethoxyxanthone, 1,8-dihydroxy-4,5,6-trimethoxyxanthone and augustin A (1,8-dihydroxy-4,6,7-trimethoxyxanthone). Structural characterization of all compounds was established on the basis of spectroscopic methods, mainly 1D and 2D nuclear magnetic resonance (NMR) and comparison with literature data. The antioxidant activity of compounds was tested through a thin layer chromatography (TLC) bioautography assay using 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH·) as detection reagent. All tested compounds were more active (DL < 0.13-0.03 µg) than Trolox (DL < 0.15 µg), used as reference standard.

Antidiarrheal mechanism and ionic profile of Carpolobia lutea ethanolic stem-bark extract in rats.

BACKGROUND: The stem-bark extract of Carpolobia lutea (Polygalaceae), used in ethno-medicine as anti-diarrhea was pharmacologically evaluated. This was the first report of assessment of the ethanolic stem extract (ESE), of C. lutea as anti-diarrhoeal agent in rats. The anti-diarrhoeal effects, acute toxicity and ionic profile are investigated and reported. MATERIALS AND METHODS: The acute toxicity was established using Lock's method. The anti-diarrhoeal effects were demonstrated using castor oil-induced diarrheal and fluid accumulation and its effect on normal intestinal transit. The mechanism elucidated using yohimbine, isosorbide dinitrate, and diphenoxylate. The elemental and ionic profile of ESE was established using inductively coupled argon-plasma emission spectrometer and potentiometric titration respectively. The finger print of ESE was revealed by Jasco (Tokyo, Japan), HPLC and active compounds by phytochemical screening using standard procedure. RESULTS: The LD50 obtained is 866.025 mg/kg (i.p). The doses of 43.3, 86.6, and 173.2 mg/kg of ESE showed inhibition of castor oil-induced diarrheal (p<0.05 -0.001). The most abundant cations in the extract are potassium and phosphorus (1.00 ±0.01 and 0.80 ± 0.030 mg/g respectively); while the most abundant anions are phosphate and sulphate (33.50±7.09 and 7.19±3.29 mg/g respectively). The HPLC fingerprint of ESE revealed UV spectra of biomolecules. Phytochemical screening revealed presence of saponins, polyphenols and glycosides. CONCLUSION: These investigations indicate presence of bioactive and elemental substances which could play major role in diarrheal management. This investigation justifies the use of stem-bark of C. lutea in illicit gin (akpatashi), among the Effiks in Nigeria as antidiarrheal.

Two new diphenylmethyl-substituted xanthones from Seicuridaca longepedunculata.

Two new diphenylmethyl-substituted xanthones, named muchimangins K (1) and L (2), have been isolated from the roots of Securidaca longepedunculata (Polygalaceae) collected in the Democratic Republic of Congo. Their structures were established by analyses of the spectral data, including 2D NMR spectra, to be 1 ,3,6,8-tetrahydroxy-2.5-dimethoxy-4-[ -(2,4,5-trimethoxyphenyl)-1-phenylmethyl]xanthone (1) and 1,3,6-trihydroxy-4,7-dimethoxy-2-[1-(2,4,5-trimethoxyphenyl)- 1-phenylmethyl]xanthone (2).

A new xanthone from Moutabea guianensis Aubl.

The ethyl acetate extract of the roots of Moutabea guianensis gave 1,6-dihydroxy-4,7,8-trimethoxy-9H-xanthen-9-one (1), a new xanthone. The isolation was accomplished by column chromatography on silica gel and the structural elucidation of this compound was established by spectroscopic analyses including 1D and 2D NMR and HRESIMS.

Protective effect of Basella alba and Carpolobia alba extracts against maneb-induced male infertility.

CONTEXT: Male infertility is one of the leading causes of social frustration and marginalization, mainly in the developing world. It is attributed to many factors including exposure to agropesticides such as manganese ethylenebis (dithiocarbamate) (maneb), which is one of the most frequently used fungicides in Cameroon. Previous reports support efficiency of some medicinal plants commonly used in Cameroonian folk medicine for the treatment of this disorder. OBJECTIVE: The present study was aimed at assessing the protective effect of extracts from selected plant species, namely Basella alba L. (Basellaceae) (MEBa) and Carpolobia alba G. Don (Polygalaceae) (AECa), in alleviating the maneb-induced impairment of male reproductive function in Wistar albino rats. MATERIALS AND METHODS: The rats were treated with vehicle, plant extract (MEBa or AECa), maneb and maneb plus plant extract, respectively, and their fertility was assessed. Animals were thereafter sacrificed and organs (liver, kidneys and reproductive organs) were dissected out and weighed. Serum androgens together with alanine aminotransferase, liver glutathione and thiobarbituric acid reactive species (TBARS) were also measured. RESULTS AND DISCUSSION: From this study, both plant extracts stimulated testosterone and improved fertility. Administration of MEBa plus maneb prevented fertility reduction by maneb and minimized the inhibitory effect of maneb on testosterone levels. AECa also improved fertility of the maneb-exposed rats, though without restoring testosterone levels, and other investigated parameters remained unaffected by different treatments. CONCLUSION: These findings emphasized the beneficial effects of B. alba and C. alba extracts on male fertility, and suggest their protective effect against maneb-induced toxicity in male reproductive function.

Effect of ethanolic extract of Carpolobia lutea G. Don (polygalaceae) root on learning and memory in CD1 mice.

Carpolobia lutea, commonly called cattle stick or poor man's candle, is used by traditional herbalists in eastern Nigeria to treat 'madness'. It has a reported analgesic and anti-nociceptive effect. The effect of its ethanolic root extract on learning and memory was investigated. Thirty mice were divided into three groups of ten each. One group of mice served as the control and was given normal saline (p.o.) while the other two groups were given acute low dose (1500mg/kg, p.o.) and high dose (2500mg/kg, p.o.) (LD50 3338.83mg/kg). The effect of the extract on cognitive memory was investigated using the Novel Object recognition task (NORT) while the effect on visuospatial learning and memory was studied using the Morris Water maze (MWM). The results obtained in the NORT show that the index of habituation was significantly lower following acute treatment with a low dose of C. lutea extract compared to control. However, the index of habituation did not differ following treatment with a high dose of C. lutea compared to control but it was higher compared to the low dose. Following treatment with a low dose of the extract, the index of discrimination was significantly higher compared to control. The index of discrimination in the high dose treatment group did not differ from control, but it was lower compared to the low dose treatment. This indicated that there was improved cognitive memory only in the low dose treatment group. In the MWM there was no significant difference in swim latency during Acquisition and Reversal training. There also was no significant difference in quadrant duration during probe trial. The swim latency during the visible platform test showed that all mice used had good visual acuity. Therefore, the ethanolic extract of C. lutea root enhanced cognitive memory. However it did not affect visuospatial learning and memory.

Pollen viability of Polygala paniculata L. (Polygalaceae) using different staining methods.

Polygala paniculata L. is a medicinal plant that grows in the Brazilian Atlantic coast, known as 'barba-de-São-João', 'barba-de-bode', 'vassourinha branca', and 'mimosa'. In this study, pollen viability was estimated by three different staining methods: 2% acetic orcein, 2% acetic carmine, and Alexander's stain. The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol: acetic acid (3:1) for 24 hours, then stored in ethanol 70% under refrigeration. Six slides per plant, two for each stain, were prepared by squashing, and 300 pollen grains per slide were analyzed. Pollen viability was high (> 70%) for most accessions of P. paniculata using the Alexander's stain, which proved the most adequate method to estimate pollen viability.

Antimicrobial activity of Carpolobia lutea extracts and fractions.

Carpolobia lutea (G. Don) (Polygalaceae) is a tropical medicinal plant putative in traditional medicines against gonorrhea, gingivitis, infertility, antiulcer and malaria. The present study evaluated the antimicrobial, antifungal and antihelicobacter effects of extracts C. lutea leaf, stem and root. The extracts were examined using the disc-diffusion and Microplates of 96 wells containing Muller-Hinton methods against some bacterial strains: Eschericia coli (ATCC 25922), E. coli (ATCC10418), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Staphyllococus aureus (ATCC 6571), Enterococcus faecalis (ATCC 29212) and Bacillus subtilis (NCTC 8853) and four clinical isolates: one fungi (Candida albican) and three bacteria (Salmonella, Sheigella and staphylococcus aureus). The Gram-positive bacteria: Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 19659) and the Gram-negative bacteria: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Cândida albicans (ATCC 18804) and Helicobacter pylori (ATCC 43504). Some of these extracts were found to be active against some tested strains but activity against H. pylori was >1000mg/ml and good fungistatic activity against C. albican. The MIC against C. albican is in the order n-HF > CHF > ETF= EAF.The order of potency of fraction was the ethanol root > n-HF leaf > ethanol fraction stem > chloroform fraction leaf = ethyl acetate fraction leaf. Polyphenols were demonstrated in ethanol fraction, ethyl acetate fraction, crude ethyl acetate extract and ethanol extract, respectively. These polyphenols isolated may partly explain and support the use of C. lutea for the treatment of infectious diseases in traditional Ibibio medicine of Nigeria.