Anandenanthera colubrina (Vell.) Brenan as an inhibitor of HIV-1 BaL infection.

We reported the in vitro anti-HIV-1 activity, cytotoxicity, cytokines expression and chemical profile of Anadenanthera colubrina. Cytotoxicity was evaluated on TZM-bl, HL2/3 cells and macrophages. Anti-HIV-1 activity was determined by Luciferase assay (TZM-bl cells) and by HIV-p24 quantification (macrophages) assessed by ELISA. TZM-bl and HL2/3 cells were used to determine cell-cell fusion inhibition. Cytokines expression was assessed by ELISA. Chemical composition was determined by Gas Chromatography Coupled to Mass Spectrometry. At 66.6 µg/mL, the extract maintained the cell viability above 90%. At 33.28 µg/mL, the extract reduced 82.8% of HIV-1 infection (TZM-bl cells) and HIV-p24 expression (macrophages). The extract inhibited approximately 70% of TZM-bl and HL2/3 cells fusion. Extract did't induce inflammatory response. Phytochemical analysis showed presence of flavonoid, phenolic acids, fatty acids and sugars. This is the first study presenting the anti-HIV effect of A. colubrina, showing low cytotoxicity and no inflammatory stimuli, important requirements for a microbicide development.

Anadenanthera colubrina (Fabaceae) logs in the Atlantic Forest biome: first host plant for Thoracibidion lineatocolle (Col: Cerambycidae) and a new host for Temnopis megacephala (Col: Cerambycidae) / Troncos de Anadenanthera colubrina (Fabaceae) no bioma da Mata Atlântica: primeira planta hospedeira de Thoracibidion lineatocolle (Col: Cerambycidae) e uma nova planta hospedeira para Temnopis megacephala (Col: Cerambycidae)

Abstract Wood-boring beetles develop in live trees and dead wood, performing ecological services such as decomposition and regulation of forest resources. Species of the Cerambycidae family, widely distributed in the world, bore into the trunks of trees and dead wood in native and cultivated areas. The objective is to report the first host plant for Thoracibidion lineatocolle (Thomson, 1865) (Coleoptera: Cerambycidae) and a new host plant for Temnopis megacephala (Germar, 1824) (Coleoptera: Cerambycidae) in the Brazilian Atlantic Forest biome. Three logs, with one-meter-long by 20 cm in diameter, were cut from the trunk of a healthy Anadenanthera colubrina (Fabaceae) tree in October 2013 and tied in the understory at 1.5m high in the Rio Doce State Park, Minas Gerais State, Brazil. The logs, exposed in the forest, were each removed after 40, 80 and 120 days and stored individually in a cardboard box in the "Laboratório de Campo do Projeto de Ecologia de Longa Duração (PELD-CNPq)" in the Rio Doce State Park. A total of 94 individuals of T. lineatocolle and 228 of T. megacephala emerged from the A. colubrina logs. This is the first report of a host plant for T. lineatocolle and a new host plant for T. megacephala.

Resumo Besouros broqueadores se desenvolvem em árvores vivas e madeira morta, realizando serviços ecológicos como decomposição e regulação de recursos da floresta. Espécies da família Cerambycidae, amplamente distribuídas no mundo, perfuram o caule de árvores e madeira morta em áreas nativas e cultivadas. O objetivo é relatar a primeira planta hospedeira de Thoracibidion lineatocolle (Thomson, 1865) (Coleoptera: Cerambycidae) e uma nova planta hospedeira para Temnopis megacephala (Germar, 1824) (Coleoptera: Cerambycidae) no bioma da Mata Atlântica brasileira. Três toras, com um metro de comprimento por 20 cm de diâmetro, foram cortadas de uma árvore sadia de Anadenanthera colubrina (Fabaceae) em outubro de 2013 e amarradas no sub-bosque a 1,5m de altura no Parque Estadual do Rio Doce, estado de Minas Gerais, Brasil. As toras, expostas na floresta, foram removidas, cada uma, após 40, 80 e 120 dias e armazenadas, individualmente, em caixas de papelão no "Laboratório de Campo do Projeto de Ecologia de Longa Duração (PELD-CNPq)" no Parque Estadual do Rio Doce. Um total de 94 indivíduos de T. lineatocolle e 228 de T. megacephala emergiu das toras de A. colubrina. Esse é o primeiro registro de uma planta hospedeira para T. lineatocolle e o de uma nova planta hospedeira para T. megacephala.

Active Flavonoids from Colubrina greggii var. greggii S. Watson against Clinical Isolates of Candida spp.

Candida albicans is the most commonly implicated agent in invasive human fungal infections. The disease could be presented as minimal symptomatic candidemia or can be fulminant sepsis. Candidemia is associated with a high rate of mortality and high healthcare and hospitalization costs. The surveillance programs have reported the distribution of other Candida species reflecting the trends and antifungal susceptibilities. Previous studies have demonstrated that C. glabrata more frequently presents fluconazole-resistant strains. Extracts from Mexican plants have been reported with activity against pulmonary mycosis, among them Colubrina greggii. In the present study, extracts from the aerial parts (leaves, flowers, and fruits) of this plant were evaluated against clinical isolates of several species of Candida (C. albicans, C. glabrata, C. parapsilosis, C. krusei, and C. tropicalis) by the broth microdilution assay. Through bioassay-guided fractionation, three antifungal glycosylated flavonoids were isolated and characterized. The isolated compounds showed antifungal activity only against C. glabrata resistant to fluconazole, and were non-toxic toward brine shrimp lethality bioassay and in vitro Vero cell line assay. The ethyl acetate and butanol extracts, as well as the fractions containing the mixture of flavonoids, were more active against Candida spp.

Anadenanthera colubrina (Fabaceae) logs in the Atlantic Forest biome: first host plant for Thoracibidion lineatocolle (Col.: Cerambycidae) and a new host for Temnopis megacephala (Col.: Cerambycidae).

Wood-boring beetles develop in live trees and dead wood, performing ecological services such as decomposition and regulation of forest resources. Species of the Cerambycidae family, widely distributed in the world, bore into the trunks of trees and dead wood in native and cultivated areas. The objective is to report the first host plant for Thoracibidion lineatocolle (Thomson, 1865) (Coleoptera: Cerambycidae) and a new host plant for Temnopis megacephala (Germar, 1824) (Coleoptera: Cerambycidae) in the Brazilian Atlantic Forest biome. Three logs, with one-meter-long by 20 cm in diameter, were cut from the trunk of a healthy Anadenanthera colubrina (Fabaceae) tree in October 2013 and tied in the understory at 1.5m high in the Rio Doce State Park, Minas Gerais State, Brazil. The logs, exposed in the forest, were each removed after 40, 80 and 120 days and stored individually in a cardboard box in the "Laboratório de Campo do Projeto de Ecologia de Longa Duração (PELD-CNPq)" in the Rio Doce State Park. A total of 94 individuals of T. lineatocolle and 228 of T. megacephala emerged from the A. colubrina logs. This is the first report of a host plant for T. lineatocolle and a new host plant for T. megacephala.

In vitro and in vivo evaluation of anti-inflammatory activity and free radical scavenging potential of leaves extract from Anadenanthera colubrina.

The present study highlights the antioxidant and anti-inflammatory activities from Anadenanthera colubrina leaves ethanolic extract (EEAc) and its phytochemical profile. The chemical profile of EEAc was determined and in vitro free radical scavenging potential, cell viability in RAW 264.7 and in vitro and in vivo anti-inflammatory activity were evaluated. The analysis of EEAc showed several phenolic compounds such as tannins and phenolic acids. The high antioxidant potential observed is possibly due to its high phenolic content. The EEAc (500 mg kg-1) showed an in vivo effect at the same level of dexamethasone; in vitro, at 50 µg mL-1, inhibited approximately 80% of nitric oxide production concentration, showed an inhibition of more than 50% of TNF-α production and presented high cell viability. The results show that A. colubrina leaves are an important source of phytochemicals that possesses antioxidant and anti-inflammatory properties.

Two new bioactive triterpenoids from the roots of Colubrina asiatica.

Two new ceanothane triterpenes, 3,7-O,O-dibenzoyl ceanothic acid methylester (1) and 3-O-acetyl-7-O-benzoyl ceanothic acid methylester (2), along with nine known compounds (3-11), were isolated from the roots of Colubrina asiatica. The isolated compounds were identified by spectroscopic evidence. Compounds 1 and 2 showed antimalarial activity against Plasmodium falciparum with IC50 values of 4.67 and 3.07 µg/mL, respectively. Compound 2 also showed antimycobacterial activity against Mycobacterium tuberculosis (MIC 6.25 µg/mL). In addition, compounds 1, 2, 10 and 11 showed cytotoxicity against three cancer cell lines (KB, NCI-H187 and MCF-7) with IC50 values ranging from 8.32 to 46.72 µg/mL.

Novel topical skin hydration agent containing Anadenanthera colubrina polysaccharide-standardized herbal preparation.

BACKGROUND: Hydration is an important factor to promote skin barrier function, metabolism, and appearance. In this process, the presence of aquaglyceroporins, envelope and lipid synthesis, and metabolism proteins are essential to provide greater corneocyte cohesion and to form a barrier avoiding transepidermal water loss. OBJECTIVE: We evaluated the effects of a new topical pigment-free agent containing an Anadenanthera colubrina polysaccharide-rich dermocosmetic preparation (ACP) on the aquaporin-3 (AQP-3), filaggrin (FLG), involucrin (INV), glucocerebrosidase (GBA), and elongation of very-long-chain fatty acid (ELOVL) proteins production in skin human fragments, as well as on the transepidermal water loss in a double-blind placebo-controlled clinical trial. METHODS: AQP3, FLG, INV, GBA, and ELOVL3 levels were measured by immunofluorescence analysis in human skin explants. Clinical trial was conducted to evaluate the effects of ACP 1% and ACP 3% on the transepidermal water loss (TEWL). RESULTS: Image and statistical analysis showed that ACP 3% significantly increased at 90% the expression of AQP3. Similarly, ACP 3% was able to promote a significant increase of 68% and 51% in FLG and INV, respectively. ACP 3% produced no effects on the GBA and ELOVL3 proteins. Transepidermal water loss was significantly reduced in human volunteers under treatment with ACP 1% and ACP 3%. CONCLUSION: ACP reduced transepidermal water loss in a clinical trial, promoting human skin hydration. These effects were related to modulation of the AQP3, FLG, and INV as evidenced by immunofluorescence assay. This way, A colubrina polysaccharide-rich phytopharmaceutical preparation is an effective additive product to skin hydration.

Interrogation of ethnomedicinal plants for synthetic lethality effects in combination with deficiency in the DNA repair endonuclease RAD1 using a yeast cell-based assay.

ETHNOPHARMACOLOGICAL RELEVANCE: Plant materials used in this study were selected based on the ethnobotanical literature. Plants have either been utilized by Thai practitioners as alternative treatments for cancer or identified to exhibit anti-cancer properties. AIM OF THE STUDY: To screen ethnomedicinal plants using a yeast cell-based assay for synthetic lethal interactions with cells deleted for RAD1, the yeast homologue of human ERCC4 (XPF) MATERIALS AND METHODS: Ethanolic extracts from thirty-two species of medicinal plants utilized in Thai traditional medicine were screened for synthetic lethal/sick interactions using a yeast cell-based assay. Cell growth was compared between the parental strain and rad1∆ yeast following exposure to select for specific toxicity of plant extracts. Candidate extracts were further examined for the mode of action using genetic and biochemical approaches. RESULTS: Screening a library of ethanolic extracts from medicinal plants identified Bacopa monnieri and Colubrina asiatica as having synthetic lethal effects in the rad1∆ cells but not the parental strain. Synthetic lethal effects for B. monneiri extracts were more apparent and this plant was examined further. Genetic analysis indicates that pro-oxidant activities and defective excision repair pathways do not significantly contribute to enhanced sensitivity to B. monneiri extracts. Exposure to B. monneiri extracts resulted in nuclear fragmentation and elevated levels of ethidium bromide staining in rad1∆ yeast suggesting promotion of an apoptosis-like event. Growth inhibition also observed in the human Caco-2 cell line suggesting the effects of B. monnieri extracts on both yeast and human cells may be similar. CONCLUSIONS: B. monneiri extracts may have utility in treatment of colorectal cancers that exhibit deficiency in ERCC4 (XPF).

Chemical constituents and biological activities from branches of Colubrina asiatica.

Sixteen compounds were isolated from a Thai medicinal plant, Colubrina asiatica. The isolated compounds were elucidated on the basis of spectroscopic methods (IR, 1D and 2D NMR) as six triterpene acids (1-6), five steroids (7-11), one benzoic acid derivative (12), two peptides (13 and 14), one sesquiterpenoid (15) and one jujubogenin (16). Compounds 3 and 10 showed antimalarial activity against Plasmodium falciparum. Compound 5 showed antimycobacterial activity. Moreover, compounds 3, 5, 6, 10 and 14 exhibited weak cytotoxicity against cancer cell lines. Compounds 1-15 have been isolated for the first time from this plant.

Elevated International Normalized Ratio in a Patient Taking Warfarin and Mauby: A Case Report.

We describe a 70-year-old Haitian man who had been taking warfarin for 5 years for atrial fibrillation and pulmonary hypertension. This patient had his international normalized ratio (INR) checked in the pharmacist-run anticoagulation clinic and was followed monthly. Prior to the interaction, his INR was therapeutic for 5 months while taking warfarin 10.5 mg/d. The patient presented with an INR > 8.0. Patient held 4 days of warfarin and restarted on warfarin 8.5 mg/d. Two weeks later, his INR was 2.5. After continuing dose, patient presented 2 weeks later and INR was 4.8. Upon further questioning, the patient stated he recently began ingesting mauby. Mauby is a bitter dark liquid extracted from the bark of the mauby tree that is commonly used in the Caribbean population as a folk remedy with many health benefits. This case report illustrates that mauby may have a probable drug-herb interaction (Naranjo Algorithm Score of 6) when given with warfarin. There is a lack of published literature and unclear information on the Internet describing the interaction of mauby and warfarin. Health professionals should be cautious regarding interactions between warfarin and mauby until the interaction is fully elucidated.

Chemometrics-enhanced high performance liquid chromatography-ultraviolet detection of bioactive metabolites from phytochemically unknown plants.

This work describes the use of Colubrina greggii as a model to investigate the use of chemometric analysis combined with data from a leishmanicidal bioassay, using Principal Component Analysis (PCA) and Orthogonal Projections to Latent Structures (O-PLS), to detect biologically active natural products in crude extracts from plants having little or no phytochemical information. A first analysis of the HPLC-UV profiles of the extract and its semi-purified fractions using both Principal Component Analysis (PCA) and Orthogonal Partial Least Squares (O-PLS) indicated that the components at tR 48.2, 48.7, 51.8min correlated with the variation in bioactivity. However, a further O-PLS analysis of the HPLC-UV profiles of fractions obtained through a final semi-preparative HPLC purification showed two components at tR 48.7 and 49.5min which correlated with the variation of the bioactivity in a high performance predictive model, with high determination coefficient, high correlation coefficient values (R(2) and Q(2)=0.99) and a low root mean square error (RMSE=0.018). This study demonstrates that the association of chemometric analysis with bioassay results can be an excellent strategy for the detection and isolation of bioactive metabolites from phytochemically unknown plant crude extracts.

A unifying study of phenotypic and molecular genetic variability in natural populations of Anadenanthera colubrina var. cebil from Yungas and Paranaense biogeographic provinces in Argentina.

Anadenanthera colubrina var. cebil is a discontinuously distributed native tree species in South American subtropical forests. Thirteen quantitative traits and eight nuclear microsatellite loci were examined in individuals from two biogeographic provinces of Argentina to determine the number and composition of genetically distinguishable groups of individuals and explore possible spatial patterns of the phenotypic and genetic variability. Means of reproductive traits were higher in the Yungas than in the Paranaense biogeographic province, whereas five out of eight nonreproductive quantitative traits showed higher mean values in the latter. Variance coefficients were moderate, and there were significant differences between and within provinces. Three clusters were defined based on spatial model for cluster membership for quantitative traits. One cluster grouped the individuals from the Paranaense biogeographic province whereas the individuals from the Yungas biogeographic province grouped regarding its population of origin. Parameters of molecular genetic variability showed higher values in the Yungas than in the Paranaense biogeographic province. Observed heterozygosity was lower than expected heterozygosity in both biogeographic provinces, indicating an excess of homozygosity. The homozygosity test by Watterson and the exact test by Slatkin suggested diversifying selection for locus Ac41.1. Bayesian clustering spatial model for microsatellites loci data were performed for both all loci and for all loci excluding locus Ac41.1. In both analyses two clusters were inferred. Analysis of molecular variance revealed similar results for all genotypes and for all genotypes defined excluding locus Ac41.1. Most of the total variance is attributable to genetic variation within clusters. The presence of homogeneous clusters was detected for both the phenotypic and molecular genetic variability. Two Bayesian clustering analyses were performed according to molecular genetic data, and two clusters were inferred. Individuals were assigned to their provinces of origin. Genetic molecular variation was higher in the populations of the Yungas biogeographic province which translates into highly qualified populations for conservation. Populations from the Paranaense biogeographic province showed the highest mean value of number of seeds per fruit making them valuable as well with regard to the exploitation of management strategies as a means to recover the impacted areas where these populations are located.

Antioxidant and orofacial anti-nociceptive activities of the stem bark aqueous extract of Anadenanthera colubrina (Velloso) Brenan (Fabaceae).

The anti-nociceptive and antioxidant activities of the Anadenantheracolubrina stem bark aqueous extract (AEAC) were investigated. AEAC (30 µg/mL) reduced 94.8% of 2,2-diphenyl-1-picrylhydrazyl radical and prevented 64% (200 µg/mL) of lipid peroxidation caused by 2,2'-azobis(2-methylpropionamidine) dihydrochloride-induced peroxyl radicals. AEAC treatment (200 and 400 mg/kg) significantly (p < 0.001) reduced mice orofacial nociception in the first (61.4% and 62.6%, respectively) and second (48.9% and 61.9%, respectively) phases of the formalin test. Nociception caused by glutamate was significantly (p < 0.001) reduced by up to 79% at 400 mg/kg, while 56-60% of the nociceptive behaviour induced by capsaicin was significantly inhibited by AEAC (100-400 mg/kg). Mice treated with AEAC did not show changes in motor performance in the Rota-rod apparatus. It appears that AEAC is of pharmacological importance in treating pain due to its anti-nociceptive effects, which were shown to be mediated by central and peripheral mechanisms.

Uso de protetor físico na semeadura direta para recuperação de áreas degradadas / Use of physical protector in tillage for recovery of degraded areas

A semeadura direta apresenta-se como uma técnica viável economicamente e de fácil implantação para recuperação de áreas degradadas, no entanto problemas na emergência e crescimento inicial das plântulas são fatores limitantes para sua aplicação. Portanto, o presente trabalho teve como objetivo avaliar a eficiência do uso de protetor físico na emergência e crescimento inicial de três espécies arbóreas, Anadenanthera colubrina, Leucaena leucocephala e Sena pendula. Para a proteção das espécies foram utilizados copos plásticos de 400 mL com o fundo removido, sendo que, no período de 45 dias foram avaliadas a emergência, sobrevivência e altura das plântulas. Observou-se que, a utilização de protetor físico não foi significativo para a sobrevivência e emergência das espécies estudadas, porém o crescimento inicial em altura das plantas apresentou melhores resultados nas sementes com protetores quando comparadas às sem proteção, para as espécies L. leucocephala e A. colubrina. Pode-se concluir que, dispensa-se o uso de protetor para aumentar a emergência e sobrevivência das espécies em estudo, mas este é indicado quando deseja-se obter um maior incremento em altura.

The direct seeding is presented as an economically viable technique for easy deployment and recovery of degraded areas, however problems in the emergence and early seedling growth are limiting factors for their application. Therefore, this study aimed to evaluate the effectiveness of using physical protector in the emergence and early growth of three tree species, Anadenanthera colubrine, Leucaena leucocephala e Sena pendula. For the protection of the species used were 400 mL plastic cups with the bottom removed, and, within 45 days were examined emergence, survival and seedling height. It was observed that the use of physical protector was not significant for the survival and emergence of the species, but the initial growth of plant height showed the best results with floating seeds when compared to no protection for the species L. leucocephala and A. colubrina. It can be concluded that dismissal is the use of protective emergency and to increase the survival of the species studied, but this is indicated when it is desired to obtain a greater increase in height.

Expression of differential genes involved in the maintenance of water balance in human skin by Piptadenia colubrina extract.

BACKGROUND: Hydration and integrity of the stratum corneum (SC) is an important determinant of skin appearance, metabolism, mechanical properties, and barrier function. The presence of aquaglyceroporins and envelope proteins are crucial to provide greater corneocyte cohesion to keep water and other moisturizers in the skin. AIMS: In this study, we evaluated the ability of Piptadenia colubrina, a plant native of South American rain forests, in the expression of genes involved in skin capacitance and SC integrity. METHODS: The expression of genes for aquaporin-3 (AQP3), loricrin, involucrin (INV), and filaggrin (FLG) was measured by real-time PCR, using an in vitro model of human keratinocytes incubated with concentrations of 2.5, 5, 10, and 20 mg/mL of a hydroglycolic extract of P. colubrina (HEPC). The amount of AQP3 protein was also tested by immunohistochemistry in human skin explants. Clinical trials were conducted to evaluate the effects of a gel-cream containing HEPC on the glycerol index and skin capacitance. RESULTS: Hydroglycolic extract of P. colubrina increased both the expression and immunoreactivity of AQP3 in cultured keratinocytes and human skin explants. The gene induction to envelope proteins FLG and INV was also observed after cell incubation with HEPC. Skin capacitance was significantly improved in human volunteers under treatment with HEPC-containing cream. CONCLUSIONS: The extract of P. colubrina promotes cellular hydration and induces gene expression of envelope proteins providing greater corneocyte cohesion to keep water and other moisturizers in the skin and an appropriate epidermal adhesion. The in vitro findings were clinically confirmed and encourage the clinical use of this compound in skin care products.

The control of hypertension by use of coconut water and mauby: two tropical food drinks.

In this study, the authors investigated the effect of regular consumption of two tropical food drinks, coconut (Cocos nucifera) water and mauby (Colubrina arborescens), on the control of hypertension. Twenty-eight hypertensive subjects were assigned to four equal groups and their systolic and diastolic blood pressures recorded for two weeks before and then for another two weeks while receiving one of four interventions. One group (the control) received bottled drinking water, the second group received coconut water, the third received mauby and the fourth group, a mixture of coconut water and mauby. Significant decreases in the mean systolic blood pressure were observed for 71%, 40% and 43% respectively of the groups receiving the coconut water, mauby and the mixture (p < or = 0.05). For these groups, the respective proportions showing significant decreases in the mean diastolic pressure were 29%, 40% and 57%. For the group receiving the mixture, the largest decreases in mean systolic and mean diastolic pressure were 24 mmHg and 15 mmHg respectively; these were approximately double the largest values seen with the single interventions.

The control of hypertension by use of coconut water and mauby: two tropical food drinks

In this study, the authors investigated the effect of regular consumption of two tropical food drinks, coconut (Cocos nucifera) water and mauby (Colubrina arborescens), on the control of hypertension. Twenty-eight hypertensive subjects were assigned to four equal groups and their systolic and diastolic blood pressures recorded for two weeks before and then for another two weeks while receiving one of four interventions. One group (the control) received bottled drinking water, the second group received coconut water, the third received mauby and the fourth group, a mixture of coconut water and mauby. Significant decreases in the mean systolic blood pressure were observed for 71, 40 and 43 respectively of the groups receiving the coconut water, mauby and the mixture (p < or = 0.05). For these groups, the respective proportions showing significant decreases in the mean diastolic pressure were 29, 40 and 57. For the group receiving the mixture, the largest decreases in mean systolic and mean diastolic pressure were 24 mmHg and 15 mmHg respectively; these were approximately double the largest values seen with the single interventions

Population genetic structure of two rare tree species (Colubrina oppositifolia and Alphitonia ponderosa, Rhamnaceae) from Hawaiian dry and mesic forests using random amplified polymorphic DNA markers.

Hawaiian dry and mesic forests contain an increasingly rare assemblage of species due to habitat destruction, invasive alien weeds and exotic pests. Two rare Rhamnaceae species in these ecosystems, Colubrina oppositifolia and Alphitonia ponderosa, were examined using random amplified polymorphic DNA (RAPD) markers to determine the genetic structure of the populations and the amount of variation relative to other native Hawaiian species. Relative variation is lower than with other Hawaiian species, although this is probably not a consequence of genetic bottleneck. Larger populations of both species contain the highest levels of genetic diversity and smaller populations generally the least as determined by number of polymorphic loci, estimated heterozygosity, and Shannon's index of genetic diversity. Populations on separate islands were readily discernible for both species as were two populations of C. oppositifolia on Hawai'i island (North and South Kona populations). Substructure among Kaua'i subpopulations of A. ponderosa that were ecologically separated was also evident. Although population diversity is thought to have remained at predisturbance levels, population size continues to decline as recruitment is either absent or does not keep pace with senescence of mature plants. Recovery efforts must focus on control of alien species if these and other endemic dry and mesic forest species are to persist.