RESUMO -O objetivo deste trabalho foi investigar a diversidade, a estrutura genética e o tamanho efetivo, retido em um banco de germoplasma, de duas populações de Myracrodruon urundeuva Fr. All. (aroeira), procedentes de Aramina-SP e Selvíria-MS. As populações foram avaliadas a partir da amostragem de 25 progênies de polinização aberta de cada população. De cada progênie, 17 a 20 indivíduos foram amostrados e genotipados para oito locos microssatélites. Os maiores valores para o número total de alelos (A t ), número médio de alelos por loco (A), número efetivo de alelos (A e ), heterozigosidade observada (H 0 ) e esperada (H e ) foram detectados na população Selvíria (A t = 105, A = 13,13, A e = 3,98, H 0 = 0,669 e H e = 0,749), enquanto Aramina teve A t = 94, A = 13,75, A e = 3,10, H 0 = 0,535 e H e = 0,678. A diferenciação nas frequências alélicas das duas populações, com relação ao pólen cruzado (0,159) e do óvulo (0,235), indica que cerca de 84% e 77%, respectivamente, da diversidade genética está dentro das populações. O coeficiente médio de coancestria foi maior (Selvíria Θ = 0,165, Aramina Θ = 0,169) e o tamanho efetivo médio das progênies (Selvíria N e(v) = 3,04, Aramina N e(v) = 2,69) foi menor do que o esperado em progênies de populações panmíticas (Θ = 0,125, N e(v) = 4). O tamanho efetivo total retido no banco ex situ foi estimado em 67,5 na população Selvíria e 71,1 na população Aramina, valores menores do que o requerido (N e = 150) para a conservação de populações em curto prazo. Entretanto, as duas populações apresentaram alta diversidade genética, o que as qualifica para serem utilizadas em programas de conservação e melhoramento genético da espécie, desde que seja aumentado o tamanho efetivo populacional conservado ex situ.Palavras-chave: Microssatélites, Espécie arbórea tropical e Endogamia. GENETIC DIVERSITY AND EFFECTIVE SIZE IN TWO Myracrodruon urundeuva FR. ALL., POPULATION UNDER EX SITU CONSERVATION

The eye is immunologically privileged when inflammatory responses are suppressed. One component responsible for the suppression of inflammatory responses is the blood retinal barrier, which comprises the retinal pigment epithelium. The destruction of this barrier initiates inflammation, which can affect any part of the eye. Therefore, inflammatory response is controlled by the action of anti-inflammatory mediators, among these mediators, annexin A1 (ANXA1) protein acts as a modulator of inflammation. In this study we aimed to improve the knowledge of this area by investigating how a peptide of the ANXA1 protein (ANXA1) modulates the morphology, proliferation, migration and expression of genes and proteins in human retinal pigment epithelium cells (ARPE-19). Determining how signaling pathways (NF-κB and UBC) are modulated by the ANXA1 peptide could be important for understanding the inflammatory process. ARPE-19 cells were activated by bacterial lipopolysaccharide endotoxin (LPS) and treated with ANXA1 peptide, in a concentration of 1.7μM and 33.8μM. We observed that the LPS activation diminished the levels of endogenous ANXA1 after 2h and 24h and ANXA1 peptide decreased the proliferation and re-establishes the migration of ARPE-19 cells. After using a hybridization approach, 80 differentially expressed genes were found. Five of these genes were selected (LRAT, CTGF, MAP1B, ALDH1A3 and SETD7) and all were down-regulated after treatment with the peptide. The genes CTGF and LRAT would be considered as potential molecular markers of ophthalmologic inflammation. The expression of pro-inflammatory cytokines was also decreased after the treatment, indicating the efficiency of the anti-inflammatory peptide at high concentrations, since the reduction in the levels of these mediators were observed after the treatment with ANXA1 peptide at 33.8μM. Our results suggest that the retinal pigment epithelial cells are a potential target of the ANXA1 protein and point to possible applications of the ANXA1 peptide as an innovative therapy for the treatment of ocular inflammation.

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Cervical cancer is one of the leading causes of cancer death in women worldwide, and its tumorigenesis can be influenced by the microenvironment. The anti‐inflammatory protein annexin A1 (ANXA1) has been reported to be associated with cancer progression and metastasis, suggesting that it plays a role in regulating tumour cell proliferation. Here, we examined the effect of the N‐terminal peptide Ac2‐26 of ANXA1 on the HaCaT cell line (normal) and HeLa cell line (cervical cancer) co‐cultured with endothelium cell‐conditioned medium (HMC). Treatment with Ac2‐26 decreased proliferation and increased motility of cervical cancer cells, but did not affect cellular morphology or viability. Combined HMC stimulus and Ac2‐26 treatment resulted in an increase in apoptotic HeLa cells, upregulated expression of MMP2, and downregulated expression of COX2, EP3 and EP4. In conclusion, Ac2‐26 treatment may modulate cellular and molecular mechanisms underlying cervical carcinogenesis.

-Mating system in Myracrodruon urundeuva (Anarcadiaceae): implications for conservation genetics. The aims of this study were to investigate the mating system of a fragmented population of the dioecious tropical tree Myracrodruon urundeuva Allemão, using five microsatellite loci and the mixed mating and correlated mating models. The study was conducted in the Estação Ecológica de Paulo de Farias (436 ha), where the population occupies about 142 ha. The mating system was estimated using 514 open-pollinated offspring, collected from 30 seed-trees. Estimates of the multilocus outcrossing rate confirm that the species is dioecious (t m = 1.0). Low levels of mating among relatives were detected in the population (1 -t s = 0.020). The estimate of paternity correlation (r p(m) ) indicated that offsprings were composed of mixtures of half-sibs and full-sibs, with the latter occurring at a low frequency (average of 0.148). The estimated coancestry coefficient within families (Θ = 0.147) was larger and the effective population size (N e(v) ) was lower (N e(v) = 2.98) than expected in progenies from panmictic populations (Θ = 0.125, N e(v) = 4, respectively). In terms of conservation, the results indicate that to retain an effective population size of 150, is necessary to collect seeds from at least 50 seed-trees.Key words -correlated matings, fragmentation, microsatellite, variance effective population size RESUMO -Sistema de reprodução em Myracrodruon urundeuva (Anarcadiaceae): implicações para a conservação genética. O objetivo deste estudo foi investigar o sistema de reprodução de uma população fragmentada da espécie arbórea dióica, Myracrodruon urundeuva Allemão, utilizando cinco locos microssatélites e os modelos mistos de reprodução e de cruzamentos correlacionados. O estudo foi conduzido na Estação Ecológica de Paulo de Faria (436 ha), onde a população ocupa uma área aproximada de 142 ha. Foram amostradas 514 progênies de polinização aberta coletadas de 30 árvores matrizes da população. A estimativa da taxa de cruzamento multilocus confirma que a espécie é dióica (t m = 1,0). Baixos níveis de cruzamentos entre parentes foram detectados na população (1 -t s = 0,020). A estimativa da correlação de paternidade (r p(m) ) indicou que as progênies são compostas por misturas de meios-irmãos e irmãos-completos, embora este último ocorra em menor proporção (média de 0,148). O coeficiente de coancestria dentro das progênies (Θ = 0,147) foi maior e o tamanho efetivo de variância (N e(v) = 2,98) foi menor do que o esperado em progênies de populações panmíticas (Θ = 0,125, N e(v) = 4, respectivamente). Em termos de conservação genética, os resultados indicam que para se obterem lotes de sementes com tamanho efetivo de referência de 150, é necessário coletar sementes de pelo menos 50 árvores matrizes.Palavras-chave -cruzamentos correlacionados, fragmentação, microssatélites, tamanho efetivo populacional 1.Parte da tese de doutorado da primeira autora,

BackgroundSome plants had been used in the treatment of cancer and one of these has attracted scientific interest, the Euphorbia tirucalli (E. tirucalli), used in the treatment of asthma, ulcers, warts has active components with activities scientifically proven as antimutagenic, anti-inflammatory and anticancer.MethodsWe evaluate the influence of the antitumoral fraction of the E. tirucalli latex in the larynx squamous cell carcinoma (Hep-2), on the morphology, cell proliferation and gene expression. The Hep-2 cells were cultivated in complete medium (MEM 10 %) and treated with E. tirucalli latex for 1, 3, 5 and 7 days. After statistically analyzing the proliferation of the tested cells, the cells were cultivated again for RNA extraction and the Rapid Subtractive Hybridization (RaSH) technique was used to identify genes with altered expression. The genes found using the RaSH technique were analyzed by Gene Ontology (GO) using Ingenuity Systems.ResultsThe five genes found to have differential expression were validated by real-time quantitative PCR. Though treatment with E. tirucalli latex did not change the cell morphology in comparison to control samples, but the cell growth was significantly decreased. The RaSH showed change in the expression of some genes, including ANXA1, TCEA1, NGFRAP1, ITPR1 and CD55, which are associated with inflammatory response, transcriptional regulation, apoptosis, calcium ion transport regulation and complement system, respectively. The E. tirucalli latex treatment down-regulated ITPR1 and up-regulated ANXA1 and CD55 genes, and was validated by real-time quantitative PCR.ConclusionsThe data indicate the involvement of E. tirucalli latex in the altered expression of genes involved in tumorigenic processes, which could potentially be applied as a therapeutic indicator of larynx cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1115-z) contains supplementary material, which is available to authorized users.

Dichondra repens (kidneyweed or ponysfoot), family Convolvulaceae, is a perennial plant with persistent leaves and is grown alone or in association with turfgrass in subtropical and Mediterranean regions. Because of its prostrate growth habit, it does not need to be mowed. It is also used as a potted plant for house decoration. During surveys of lawns in public gardens of the Franco-Italian Riviera conducted from 1993 to 2003, we noticed 0.1- to 0.5-cm-diameter, brownish, necrotic spots on leaves of D. repens in Antibes, Cannes, Menton, Nice, and Vallauris (France) and in Arma di Taggia, Diano Marina, Imperia, La Mortola, Ospedaletti, San Remo, and Ventimiglia (Italy). Symptoms were more intense in the spring on young leaves but lesions remained all year on older leaves. Two species of fungal pathogens were frequently isolated from these spots. One fungus produced brown, erect conidiophores with brown, pear-shaped conidia and bifid, subhyaline beaks. Conidia formed singly, were composed of 8 to 10 cells with transverse and longitudinal crosswalls, and had one to four hyaline spurs frequently longer than the conidia. Conidia measured 90 to 260 × 16 to 29 μm. The pathogen, identified as Alternaria dichondrae (1), was previously characterized in Italy, New Zealand, and Argentina. The second fungus species produced clumps of erect, brown conidiophores with hyaline, filiform conidia composed of 10 to 20 cells. These conidia measured 90 to 310 × 3 to 3.5 μm. This fungus was identified as a Cercospora sp. (2), a genus not previously reported on D. repens. For both fungi, necrotic spots similar to those observed in natural infections were obtained after spraying a suspension of mycelium and conidia onto leaves of D. repens seedlings that had two to four expanded leaves that had been pricked with a pin. The plants were maintained under high humidity. Assays of mycelium growth on agar media containing various fungicides showed that 1 ppm of pyremethanil completely inhibited the growth of A. dichondrae, whereas a mixture of 10 ppm of diethofencarb and 10 ppm of carbendazine completely inhibited Cercospora sp. growth. Telia were also observed on the lower surface of D. repens leaves, sometimes in association with disease symptoms of A. dichondrae and Cercospora sp. Disease symptoms of the rust were yellowing and curling of the leaf surface with erect petiole, whereas healthy plants were prostrate with plane leaf surfaces. The two-celled teliospores had smooth cell walls, a single germinative pore per cell, and measured 32 to 34 × 12 to 13 μm with a thin unattached pedicel. This rust fungus was consequently classified in the genus Puccinia (2), also not previously reported as a pathogen of D. repens. It is possible that Poaceae plants such as Poa pratensis grown in association with D. repens were the inoculum source. Whereas A. dichondrae and Cercospora sp. do not induce severe diseases and are not widespread, the prevalence of Puccinia sp. tends to increase over time, requiring appropriate treatments to manage infected turf grasses. References: (1) P. Gambogi et al. Trans. Br. Mycol. Soc. 65:322, 1975. (2) G. Viennot-Bourgin. Les Champignons Parasites des Plantes Cultivées, Masson ed. Paris, 1949.

Bergenia crassifolia (L.) Fritsch (elephant's ears or Siberian tea) (Saxifragaceae) is a perennial rhizomatous plant with pink flowers appearing at the end of winter. Since 1990, large, brown, and necrotic spots have been observed on numerous B. crassifolia plants at the University of Sciences in Nice, France. Spots appeared each year in the spring on newly emerged leaves and enlarged up to 1 to 3 cm in diameter during the summer, sometimes affecting more than half of the leaf surface. Leaves with spots were collected from May to November and placed in a humid atmosphere. Black, sessile, discoid conidiomata developed on the spots and exuded a pink, then brown, spore mass. When a mass was transferred onto a 1% malt agar medium, mycelium grew and then numerous, relatively spherical conidiomata (0.5 to 2.5 mm in diameter) developed and exuded a pink slimy mass, which contained many conidia. The mycelium grown at 24°C in the dark was scarce and pale, pink-beige. Under the light, the fungal culture was much darker with a fluffy mycelium and numerous conidiomata. The base of the conidiomata was dark; conidiophores were hyaline and showed little segmentation. Unicellular, cylindrical, fusiform conidia were hyaline, 5.4 to 8 μm long, and 1.4 to 1.9 μm wide. The morphology and size of conidia were comparable with previous descriptions of Pilidium concavum (Desm.) Höhn. (2,3). The ITS1-5.8S-ITS2 region of two isolates was amplified by PCR with primers PN3 and PN10 according to Mendes-Pereira et al. (1) and sequenced. The 421-nt sequence (GenBank Accession No. FM211810) was 100% identical to that of the P. concavum specimen voucher BPI 1107275 (GenBank Accession No. AY487094). P. concavum was reported to be on stored or rotting leaves or fruits of many dicotyledonous plants (2). To validate Koch's postulates, pieces of mycelium cultures with conidiomata (28 days old) were placed onto the upper surface of leaves of healthy B. crassifolia plants (10 to 12 pieces per plant). The leaf epidermis was previously wounded with a needle and a drop of melted paraffin was poured onto each piece of mycelium to prevent desiccation. Agar plugs without the fungus were placed similarly on wounded leaves of two control plants. Four inoculated and two control plants were incubated in growth chambers at either 24 or 18°C (16 h of light per day, 15,000 lx, 80% humidity). At 24°C, brown spots developed from 90% of the inoculation sites, whereas spots were observed for only 18% of the sites at 18°C. Such spots did not develop on control plants. After 2 months, healthy leaves as well as those with necrotic spots were put in humid chambers. Conidiomata formed after 4 weeks and exuded the same pink mass, which contained numerous conidia and from which the fungus was reisolated. Similar symptoms were also observed in several other locations in France and in botanical gardens in Akureyri (Iceland) and Métis (Canada), from which P. concavum was reisolated. To our knowledge, this is the first report of P. concavum on B. crassifolia. References: (1) E. Mendes-Pereira et al. Mycol. Res. 107:1287, 2003. (2) M. E. Palm. Mycologia 83:787, 1991. (3) A. Y. Rossman et al. Mycol. Prog. 3:275, 2004.