An evolutionary disruption of the buzz pollination syndrome in neotropical montane plants.

PREMISE: Under pollinator limitations, specialized pollination syndromes may evolve toward contrasting responses: a generalized syndrome with increased pollinator attraction, pollinator reward, and pollen transfer capacity; or the selfing syndrome with increased self-pollen deposition, but reduced pollinator attraction and pollen transfer capacity. The buzz-pollination syndrome is specialized to explore female vibrating bees as pollinators. However, vibrating bees become less-active pollinators at montane areas of the Atlantic Forest (AF) domain. This study investigated whether the specialized buzz-pollination syndrome would evolve toward an alternative floral syndrome in montane areas of the AF domain, considering a generalized and the selfing syndromes as alternative responses. METHODS: We utilized a lineage within the buzz-pollinated Miconia as study system, contrasting floral traits between montane AF-endemic and non-endemic species. We measured and validated floral traits that were proxies for pollinator attraction, reward access, pollen transfer capacity, and self-pollen deposition. We inferred the evolution of floral trait via phylogenetic comparative methods. RESULTS: AF-endemic species have selectively evolved greater reward access and more frequently had generalist pollination. Nonetheless, AF-endemic species also have selectively evolved toward lower pollen transfer capacity and greater self pollination. These patterns indicated a complex evolutionary process that has jointly favored a generalized and the selfing syndromes. CONCLUSIONS: The buzz pollination syndrome can undergo an evolutionary disruption in montane areas of the AF domain. This floral syndrome is likely more labile than often assumed, allowing buzz-pollinated plants to reproduce in environments where vibrating bees are less-reliable pollinators.

A multidisciplinary framework for biodiversity prediction in the Brazilian Atlantic Forest hotspot / Integrando disciplinas para a predição da biodiversidade da Floresta Atlântica no Brasil

Abstract: We briefly describe selected results from our thematic project focused on the biodiversity of the Atlantic Forest ("AF BIOTA"), which was jointly funded by FAPESP's BIOTA Program, the U.S. National Science Foundation Dimensions of Biodiversity Program, and the National Aeronautics and Space Administration (NASA). As one of the five most important hotspots of biodiversity in the world, the Atlantic Forest (AF) holds less than 16% of its vegetation cover, yet, amongst the hotspots, it still harbors one of the highest numbers of species, including endemics. By gathering specialists across multiple disciplines (biology, geology, engineering), we aimed to understand how this megabiodiversity was built through time, informing biodiversity science and conservation. Among the results, we trained 18 Master's and 26 Ph.D. students, published more than 400 peer-reviewed papers that improved our knowledge about the forest's biologic and climatic diversity and dynamics through time, developed new analytical methods, produced outreach videos and articles, and provided data to help define biodiversity conservation policies.

Resumo: Descrevemos de forma resumida resultados selecionados do nosso projeto temático com foco na biodiversidade da Floresta Atlântica ("AF BIOTA"), que foi financiado pelo BIOTA FAPESP e pelo programa "Dimensions of Biodiversity" da "U.S. National Science Foundation" e "National Aeronautics and Space Administration" (NASA). Devido à sua megabiodiversidade (que inclui várias espécies endêmicas), e por restar menos de 16% da vegetação original, a Floresta Atlântica (FA) é uma das cinco áreas mais importantes para a biodiversidade do planeta ("biodiversity hotspot"). Reunimos especialistas de diversas disciplinas (biologia, geologia, engenharia) visando compreender como essa megabiodiversidade evoluiu ao longo do tempo e fornecer informações científicas para a sua conservação. Dentre os resultados obtidos, nós formamos 18 mestres e 26 doutores, publicamos mais de 400 artigos científicos que aumentaram o conhecimento sobre a diversidade biológica e climática da FA e sua dinâmica ao longo do tempo, desenvolvemos novos métodos analíticos, produzimos material de divulgação científica e fornecemos dados para desenvolver políticas públicas de conservação da biodiversidade.

Flower morphology is correlated with distribution and phylogeny in Bertolonia (Melastomataceae), an herbaceous genus endemic to the Atlantic Forest.

Several centers of endemism have been proposed for Melastomataceae, particularly in Amazonia and Atlantic Forest. Despite the high degree of human-caused degradation in the last 500 years, the Atlantic Forest still presents some of the largest levels of diversity and endemism across all angiosperms. With several recently described species in the last decade, the knowledge on Bertolonia's distribution and morphological characterization has changed, with most new species found in northern Atlantic Forest and with different flower color patterns than the species from southern Atlantic Forest. We first tested the monophyly of the genus sampling over 85% of its species to generate a reliable phylogenetic hypothesis. Afterwards, we used Bertolonia as a model group to study distribution patterns and morphological evolution of lineages in the Atlantic Forest. Bertolonia is particularly interesting to address such questions because it is endemic to this domain, with species distributed either in the southern, central or northern portions of the Atlantic Forest. The second step of our work aimed to respond (1) Do closely related species endemic to Atlantic Forest tend to have similar distributions and share similar morphological traits? and (2) Are floral traits more conserved within clades than vegetative characters? We hypothesize that both (1) and (2) are true due to the low dispersal ability and consequent microendemic distribution of most species in the genus. Our results confirm the veracity of the proposed hypotheses. Three major groups were recovered in our analysis: marmorata, formosa and nymphaeifolia clades. Most species that occur in northern Atlantic Forest were recovered in the marmorata clade. Most species distributed in central Atlantic Forest were recovered in the formosa clade, and the ones from the southern Atlantic Forest in the nymphaeifolia clade. A similar pattern was recovered with reproductive characters reconstructed across the phylogeny. Generally, species with pink petals and cream-colored anthers with an extrorse pore occur in northern Atlantic Forest, and species with white petals and yellow anthers with an introrse or apical pore tend to occur in southern Atlantic Forest. Some vegetative characters also have evolutionary congruence and are restricted to one or few lineages, while other characters, such as the texture of the leaf blade surface, have a strong taxonomic value and are useful to identify species, but are not homologous. Our analysis also indicates that the division between southern and northern Atlantic Forest could also be related to ancient events, not only linked with recent phylogeographic patterns. Moreover, we suggest that the orientation of the anther pore in Bertolonia could be related with diversity of species and stability of its populations. In summary, we corroborate, based on the evolutionary history of Bertolonia, that closely related species endemic to Atlantic Forest tend to have similar distributions and share similar morphological floral traits.

Miconia lucenae (Melastomataceae), a new species from montane Atlantic Forest in Espírito Santo, Brazil.

We describe Miconia lucenae R.Goldenb. & Michelang., a new species from the montane Atlantic Forest in Santa Teresa in the state of Espírito Santo. Our analysis, based on three plastid spacers (atpF-atpH, psbK-psbl and trnS-trnG), one plastid gene (ndhF, not available for M. lucenae), and two nuclear ribosomal loci (nrITS and nrETS), showed that it belongs to a small clade with Miconia paradoxa (Mart. ex DC.) Triana (Minas Gerais) and M. michelangeliana R.Goldenb. & L.Kollmann (Espírito Santo). The three species in the "Paradoxa clade" can be recognized by the plants with glabrous or glabrescent branches and leaves, white petals and yellow stamens, these with the connectives not prolonged below the thecae, ventrally unappendaged, dorsally unappendaged or with a minute tooth, the latter bilobed or not, glabrous ovary, and the fruits with a persistent calyx. Miconia lucenae can be recognized, among the species in this clade, by the shrubby plants with terete young branches, short inflorescences, usually with red axes, and the 2-bracteolate, sessile, 4-merous flowers, with a ciliolate inner portion of the sepals, lanceolate petals, and 4-celled ovaries. This species can be considered as endangered according to IUCN criteria.

Systematics and relationships of Tryssophyton (Melastomataceae), with a second species from the Pakaraima Mountains of Guyana.

The systematics of Tryssophyton, herbs endemic to the Pakaraima Mountains of western Guyana, is reviewed and Tryssophyton quadrifolius K.Wurdack & Michelang., sp. nov. from the summit of Kamakusa Mountain is described as the second species in the genus. The new species is distinguished from its closest relative, Tryssophyton merumense, by striking vegetative differences, including number of leaves per stem and leaf architecture. A phylogenetic analysis of sequence data from three plastid loci and Melastomataceae-wide taxon sampling is presented. The two species of Tryssophyton are recovered as monophyletic and associated with mostly Old World tribe Sonerileae. Fruit, seed and leaf morphology are described for the first time, biogeography is discussed and both species are illustrated.

Real-time PCR detection of a 16S rRNA single mutation of Helicobacter pylori isolates associated with reduced susceptibility and resistance to tetracycline in the gastroesophageal mucosa of individual hosts.

The molecular mechanism of Helicobacter pylori resistance to tetracycline involves mutations in the primary binding site of the ribosome. A resistance or reduced susceptibility to tetracycline could be the result of single, double or triple mutations in the 16S rRNA gene of H. pylori. We investigated if the genotype was correlated to tetracycline resistance as determined phenotypically in vitro for 96 H. pylori isolates in the gastroesophageal mucosa of Venezuelan individual hosts. E-test for antimicrobial susceptibility test and real-time PCR for the detection of 16S rRNA gene mutations were performed in 96 H. pylori isolates (48 obtained from antrum, and 48 from oesophagus) from eight dyspeptic patients. In the gastric mucosa, 38 isolates were identified sensitive and 10 resistant to tetracycline by E-test, whereas 44 sensitive and 4 resistant isolates were found in the oesophagus. Real-time PCR detection of the 16S rRNA gene exhibited mutants with a single base-pair substitution (AGA926GGA) in six antrum isolates and seven oesophagus isolates, whereas only three harboured a low level of tetracycline resistance in vitro. Our results indicate that real-time PCR detection of 16S rRNA is a reliable method to classify among tetracycline-resistant genotypes and useful in patients who have experienced a first-line treatment failure with triple therapy.

Comparison of gizzard and intestinal microbiota of wild neotropical birds.

Gut bacterial communities have been shown to be influenced by diet, host phylogeny and anatomy, but most of these studies have been done in captive animals. Here we compare the bacterial communities in the digestive tract of wild birds. We characterized the gizzard and intestinal microbiota among 8 wild Neotropical bird species, granivorous or frugivorous species of the orders Columbiformes and Passeriformes. We sequenced the V4 region of the 16S rRNA gene in 94 collected samples from 32 wild birds from 5 localities, and compared bacterial communities by foraging guild, organ, locality and bird taxonomy. 16S rRNA gene-based sequencing data were examined using QIIME with linear discriminant analysis effect size (LEfSe) and metabolic pathways were predicted using PICRUSt algorism. We identified 8 bacterial phyla, dominated by Firmicutes, Actinobacteria and Proteobacteria. Beta diversity analyses indicated significant separation of gut communities by bird orders (Columbiformes vs. Passerifomes) and between bird species (p<0.01). In lower intestine, PICRUSt shows a predominance of carbohydrate metabolism in granivorous birds and xenobiotics biodegradation pathways in frugivorous birds. Gizzard microbiota was significantly richer in granivorous, in relation to frugivorous birds (Chao 1; non-parametric t-test, p<0.05), suggesting a microbial gizzard function, beyond grinding food. The results suggest that the most important factor separating the bacterial community structure was bird taxonomy, followed by foraging guild. However, variation between localities is also likely to be important, but this could not been assessed with our study design.

Point Mutations at gyrA and gyrB Genes of Levofloxacin-Resistant Helicobacter pylori Isolates in the Esophageal Mucosa from a Venezuelan Population.

The treatment of Helicobacter pylori infection is complicated by antibiotic resistance. A high levofloxacin (LVX) resistance rate was previously demonstrated in H. pylori isolates from gastric mucosa (40%) and esophagus (19%) in individual hosts of a Venezuelan population. We aimed to assess the molecular mechanisms of LVX resistance and susceptibility in isolates from the gastroesophageal mucosa, by studying point mutations in the quinolone resistance-determining region of gyrA and gyrB genes. Sequencing of gyrA and gyrB genes (N = 120) helped to identify point mutations in 60 isolates (30 from antrum and 30 from esophagus) of five dyspeptic patients. Double (Asn87Thr and Asp91Asn) and single (Asn87Ile or Asn87Thr) mutations in the gyrA gene were identified in the esophageal mucosa. These mutations have been commonly found in the stomach. Occurrence of a single (Asn87Ile) mutation was associated with high resistance (minimum inhibitory concentration ≥ 32 µg/mL) to LVX. Only a single (Ser479Gly) mutation was found in the gyrB gene in both mucosae. One patient presented isolates with no mutations in the two genes studied. Isolates with the same mutation pattern in individual hosts revealed identical genetic profiles for these genes, confirming that isolates identified in the esophageal mucosa come from isolates colonizing the stomach. Helicobacter pylori resistance to LVX in the esophagus is related to double- and single-point mutations in gyrA and gyrB genes, such as those found in the stomach. Levofloxacin should be applied with caution, because its antibiotic effect on H. pylori is decreasing in Latin America, perhaps owing to high prescription rates.

Amazon plant diversity revealed by a taxonomically verified species list.

Recent debates on the number of plant species in the vast lowland rain forests of the Amazon have been based largely on model estimates, neglecting published checklists based on verified voucher data. Here we collate taxonomically verified checklists to present a list of seed plant species from lowland Amazon rain forests. Our list comprises 14,003 species, of which 6,727 are trees. These figures are similar to estimates derived from nonparametric ecological models, but they contrast strongly with predictions of much higher tree diversity derived from parametric models. Based on the known proportion of tree species in neotropical lowland rain forest communities as measured in complete plot censuses, and on overall estimates of seed plant diversity in Brazil and in the neotropics in general, it is more likely that tree diversity in the Amazon is closer to the lower estimates derived from nonparametric models. Much remains unknown about Amazonian plant diversity, but this taxonomically verified dataset provides a valid starting point for macroecological and evolutionary studies aimed at understanding the origin, evolution, and ecology of the exceptional biodiversity of Amazonian forests.

Multiple cag genotypes of Helicobacter pylori isolates colonize the oesophagus in individual hosts in a Venezuelan population.

PURPOSE: Multiple Helicobacter pylori strains colonize and coexist in the stomach of one single patient, carrying heterogeneous distributions of cag genotypes. The oesophagus provides a niche for H. pylori colonization; however, little is known about its adaptive role. METHODOLOGY: Using PCR for cagA, cagE and virB11 genes from cag-pathogenicity island (PAI) and Etest for antimicrobial susceptibility test, we determined cag-PAI genotypes associated with H. pylori virulence, when positive cultures were matching in both the stomach and the oesophagus (96 isolates; 8 out of 80 dyspeptic patients). RESULTS: The stomach showed complete cag-PAI islands in 77 % of the isolates, whereas the oesophagus showed complete cag-PAI islands only in 44 % of the isolates. Expression of CagA and interleukin 8 correlated with inflammatory processes and histopathological changes in the stomach, but not in the oesophagus. Different cag-PAI profiles were found in both mucosae of an individual host, and at least one oesophagus profile corresponded to one profile identified in stomach. The antibiotic resistance profiles showed variability in the colonization by single or mixed H. pylori isolates in the gastric and oesophageal mucosa both intra- and inter-individuals. CONCLUSION: These results demonstrate colonization with multiple H. pylori isolates in the oesophageal mucosa, like those found in the stomach of individual hosts. H. pylori was characterized by a dominant partial island, low interleukin 8 induction with lower histopathological damage and lower antibiotic resistance, suggesting that the microenvironmental changes in individual hosts select less virulent isolates in the oesophagus than in the stomach. New approaches to ensure effective eradication therapy in multi-resistant H. pylori strains must be developed.

Evaluación in vitro de dos extractos de Aloe vera en bacterias patógenas / In vitro evaluation of two extracts of Aloe vera in pathogenic bacteria

El incremento en los mecanismos de resistencia bacteriana, ha planteado a la comunidad científica la necesidad en la búsqueda de principios activos para producir nuevos antibióticos, donde el Aloe vera se proyecta como una fuente de obtención de los mismos. El objetivo de esta investigación fue evaluar el comportamiento de dos tipos de extractos; uno de gel fresco procesado con DMSO, y otro obtenido del mesofilo por un proceso de extracción con etanol que luego fue filtrado, concentrado y liofilizado. Ambos extractos fueron evaluados con la prueba de susceptibilidad antimicrobiana por difusión en disco contra Helicobacter pylori, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus y Streptococcus mutans. El extracto etanólico del mesófilo liofilizado se ensayó en concentraciones de 0,5; 1,0; 1,5; 2,0 y 2,5 mg/ mL, con cada una de las bacterias de referencia, evidenciándose ausencia de inhibición sobre el crecimiento bacteriano en todas las concentraciones. El extracto Gel-DMSO se ensayó con las cepas H. pylori, E. coli y S. aureus; obteniendo halos de inhibición de 14, 8,5 y 8,5 mm respectivamente. La evidencia científica de la actividad antibacteriana de esta planta suele ser contradictoria, donde el procesamiento del extracto, es un factor importante en esta variabilidad. Según nuestros resultados se puede concluir que H. pylori, fue la bacteria más sensible al extracto Gel-DMSO en comparación con E. coli y S. aureus; asimismo, para futuras investigaciones, se debería desestimar el uso de extractos liofilizados diluidos y considerar otros procesos de extracción.

The increase of bacterial resistance mechanisms has created special interest in the scientific community to search for active principles for the production of new antibiotics, and the Aloe Vera plant has been considered as a potential source to obtain them. The objective of this research was to evaluate the behavior of two types of Aloe Vera extracts, one fresh gel processed with DMSO and other obtained by the mesophyll by ethanol extraction process that was filtered, concentrated and lyophilized. Both extracts were evaluated with antimicrobial susceptibility testing by disc diffusion against Helicobacter pylori, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus and Streptococcus mutans. The ethanolic extract of lyophilized mesophyll was tested in concentrations of 0.5 1.0 1.5 2.0 and 2.5 mg / ml, with each of the reference bacteria, showing no inhibition on bacterial growth in all concentrations tested. Gel-DMSO extract was tested with strains H. pylori, E. coli and S. aureus, obtaining inhibition halos of 14, 8.5 and 8.5 mm respectively. Scientific evidence of the antibacterial activity of this plant can be contradictory, and the processing of the extract is a determining factor in such variability. According to our results, we conclude that H. pylori was the bacteria most sensitive to Gel- DMSO extract compared with E. coli and S. aureus bacteria; also, we advise against the use of diluted and lyophilised extracts is in future research; rather, other alternative extraction process should be considered.

Miconia papillosperma (Melastomataceae, Miconieae): a new species from Amazonas, Brazil.

Miconia papillosperma, a new species of Melastomataceae shrubs from Northern Brazil is described and illustrated. This new species is characterized by elliptic lanceolate leaves with the only pair of secondary veins running close to the margin. It is also unique in having seeds with a papillose testa, a character until now unknown in the Miconieae. The description of this new species from a relatively well collected area near a major road north of Manaus, Amazonas, Brazil, is further evidence of our lack of knowledge on plants in many Neotropical areas.

Prevalence and Distribution of Vibrio spp. in Wild Aquatic Birds of the Southern Caribbean Sea, Venezuela, 2011-12.

Vibrio spp. are associated with waterbirds mainly in temperate latitudes. We evaluated the prevalence and distribution of Vibrio spp. from fecal samples of resident and migratory aquatic birds collected during October 2011 and March 2012 at two coastal sites in the tropical southern Caribbean Sea. We amplified DNA by PCR in 40% of samples, resulting in 47% and 36% estimated prevalence for resident and migratory birds in Cuare Wildlife Refuge, and 33% and 44% in Margarita Island, respectively. We found nontoxigenic Vibrio cholerae in Cuare Wildlife Refuge with a higher prevalence in resident birds (18%). Our PCR results for Vibrio and V. cholerae were not significantly different between sites or bird migratory status. The 16S rRNA phylogenetic analysis sequences from fecal samples from Cuare Wildlife Refuge were highly similar to V. cholerae and Vibrio vulnificus , whereas sequences from Margarita Island samples formed clusters with species related to the Harveyi clade. Our findings indicate that several species of Vibrio are common in aquatic birds along the southern Caribbean Sea and contribute to our understanding of the role of birds as possible reservoirs of potentially pathogenic bacteria.

Angiosperms and the Linnean shortfall: three new species from three lineages of Melastomataceae at one spot at the Atlantic Forest.

Three new species of Angiosperms have been found in four short collection trips to the same protected reserve-"Estação Ecológica Estadual de Wenceslau Guimarães"-and neighboring areas in the Atlantic Forest in the south of the Brazilian state of Bahia. These new species belong to three genera from three distinct lineages in the family Melastomataceae: Huberia, Meriania and Physeterostemon. The description of these species represent a good example of a Linnean shortfall, i.e., the absence of basic knowledge about the biodiversity in the area, as well as in tropical forests as a whole. The description of these probably endemic species per se is a signal that this area deserves more attention regarding research and policies, but its consequences go farther: this area has a relevant role as a phylogenetic (both genetic and morphological) stock, and thus is also valuable as a phylogenetic conservation priority.

Untangling the phylogeny of Leandra s.str. (Melastomataceae, Miconieae).

Phylogenetic studies in Melastomataceae have demonstrated the need for taxonomic rearrangements in the current classification. Nonetheless, melastomes are among the most diverse groups of plants and several cases of known artificial taxa have been observed and awaiting further resolution. The Leandra s.str. clade, with ca. 200 species, includes the majority of the taxa traditionally treated in the genus Leandra and is almost restricted to eastern Brazil. In earlier studies, some attempts have been made to infer the relationships within Leandra s.str., but the sampling was sparse and the resolution low inside the clade. Here, we attempt to provide an improved phylogenetic hypothesis for this group on which to base further studies. Specifically, we provide a comprehensive taxon sampling and attempt to infer a species tree for this group, dissecting potential noise in the phylogenetic reconstruction, such as paralogy, rogue taxa, hybridization and incomplete lineage sorting. Our data set includes 126 ingroup species (192 terminals) and four partitions (six markers). We implement the (∗)BEAST model for species tree inference and perform several simulation methods to assess model fit and to discuss potential causes for the observed patterns. Major lineages of Leandra s.str. were delineated, a strictly bifurcating species tree model seems to not account for the observed data, and hybridization is very likely an important evolutionary force in this group.

Helicobacter pylori Infection in Rural and Urban Dyspeptic Patients from Venezuela.

The goal of this work was to assess the Helicobacter pylori prevalence in a rural mestizo population and compare it to an urban population from Venezuela. The study was performed in gastric juice samples of 71 dyspeptic patients from Caracas (urban) and 39 from Tucupita (rural), in the Orinoco Delta region. Helicobacter pylori was detected by amplification of 16S rRNA, glmM, and ureA genes in 55.0% patients from urban and 87.2% from rural populations. cagA was found positive in 51% and 62% urban and rural patients, respectively. Non-H. pylori Helicobacter species were not detected in the urban population, but was found in 7.7% of patients in the rural study site. Frequency values of the 16S rRNA, glmM, and ureA genes were higher in the rural population. The odds ratio for each gene was 15.18 for 16S rRNA, 2.34 for glmM, 2.89 for ureA, and 1.53 cagA, showing significant differences except for cagA when gene frequency was compared in both populations. These results demonstrate a higher frequency of H. pylori and gastric non-H. pylori Helicobacter infection in a rural mestizo population with low hygienic standards as compared with city dwellers, representing a potential risk for the development of gastroduodenal diseases.

Effect of tumor necrosis factor-α on the intracellular Ca(2+) homeostasis in human sperm.

PROBLEM: Inflammation and genital infections promote the increase in leukocytes, pro-inflammatory cytokines, and oxygen reactive species, impairing sperm functions such as motility, capacitation, and acrosome reaction. All these functions are primarily regulated by cytoplasmic concentration of Ca(2+) ([Ca(2+) ]cyto ). This study evaluated the effect of tumor necrosis factor (TNF)-α on the [Ca(2+) ]cyto and its regulation in human sperm. METHOD OF STUDY: Sperm loaded with fura-2 were incubated with or without TNF-α (0-500 pg/mL) from 0 to 120 min. After incubation, the basal [Ca(2+) ]cyto and membrane permeability to Ca(2+) were evaluated by spectrofluorometry, before and after Ca(2+) addition to the extracellular medium. RESULTS: Without TNF-α, the addition of Ca(2+) promotes an transitory increase in [Ca(2+) ]cyto in the spermatozoa, that returns in a few minutes to a basal level, indicating calcium regulation activation. TNF-α decreases the Ca(2+) permeation and increases the basal level of [Ca(2+) ]cyto after a Ca(2+) pulse (P < 0.04); affecting calcium regulation in a way that is time and concentration dependent. TNF-α effect was partially prevented by the addition of an antioxidant (butylated hydroxytoluene) (P < 0.03). CONCLUSION: Tumor necrosis factor-α decreases membrane permeability to Ca(2+) and affects Ca(2+) regulation in sperm cells in vitro, probably via lipid peroxidation, which may explain the decrease in sperm fertilizing capacity during inflammatory and infectious processes.

Screening of Venezuelan medicinal plant extracts for cytostatic and cytotoxic activity against tumor cell lines.

There are estimated to be more than 20,000 species of plants in Venezuela, of which more than 1500 are used for medicinal purposes by indigenous and local communities. Only a relatively small proportion of these have been evaluated in terms of their potential as antitumor agents. In this study, we screened 308 extracts from 102 species for cytostatic and cytotoxic activity against a panel of six tumor cell lines using a 24-h sulphorhodamine B assay. Extracts from Clavija lancifolia, Hamelia patens, Piper san-vicentense, Physalis cordata, Jacaranda copaia, Heliotropium indicum, and Annona squamosa were the most cytotoxic, whereas other extracts from Calotropis gigantea, Hyptis dilatata, Chromolaena odorata, Siparuna guianensis, Jacaranda obtusifolia, Tapirira guianensis, Xylopia aromatica, Protium heptaphyllum, and Piper arboreum showed the greatest cytostatic activity. These results confirm previous reports on the cytotoxic activities of the above-mentioned plants as well as prompting further studies on others such as C. lancifolia and H. dilatata that have not been so extensively studied.

Low occurrence of Helicobacter DNA in tropical wild birds, Venezuela.

Enteric Helicobacter species (Helicobacter pullorum, Helicobacter pametensis, Helicobacter canadensis, Helicobacter anseris, and Helicobacter brantae) have been found in birds from temperate latitudes. We evaluated the occurrence of Helicobacter spp. in terrestrial wild birds from Venezuela. A fragment of 16S rRNA gene was amplified by PCR with Helicobacter genus-specific primers. Helicobacter spp. were detected in four of 80 fecal and in three of 42 intestinal tissue samples. Analyses of 16S and 23S rRNA gene sequences confirm for the first time the presence of Helicobacter in tropical terrestrial wild birds. However, the occurrence of Helicobacter was low, suggesting these bacteria may be uncommon in the populations we studied.

Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.

Rotavirus infection induces an increase in [Ca(2+)](cyto), which in turn may affect the distribution of the cytoskeleton proteins in the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5 h.p.i. using fluorescent phalloidin. Western blot analysis indicated that RhoA is activated between 0.5 and 1 h.p.i. Neither the phosphorylation of RhoA nor the formation of stress fibers were observed in cells infected with virions pre-treated with an anti-VP5* non-neutralizing mAb, suggesting that RhoA activation is stimulated by the interaction of the virus with integrins forming the cell receptor complex. In addition, the structure of the tubulin cytoskeleton was also studied. Alterations of the microtubules were evident starting at 3 h.p.i. and by 7 h.p.i. when microtubules were markedly displaced toward the periphery of the cell cytoplasm. Loading of rotavirus-infected cells with either a Ca(2+) chelator (BAPTA) or transfection with siRNAs to silence NSP4, reversed the changes observed in both the microfilaments and microtubules distribution, but not the appearance of stress fibers. These results indicate that alterations in the distribution of actin microfilaments are initiated early during infection by the activation of RhoA, and that latter changes in the Ca(2+) homeostasis promoted by NSP4 during infection may be responsible for other alterations in the actin and tubulin cytoskeleton.