14 research outputs found
Identification by HS-SPME/GC-MS and activity testing of Trichoderma spp. volatile organic compounds against grapevine downy mildew
No full textDowny mildew, caused by the biotrophic oomycete Plasmopara viticola, is one of the most destructive diseases of the grapevine [1]. Fungi of the genus Trichoderma are economically important biocontrol agents since they play a crucial role in plantgrowth promotion, mycoparasitism of plant pathogens and priming of plant defence. Trichoderma spp. also release a high diversity of volatile organic compounds (VOCs), which play a decisive role against plant pathogens [2]. However, the possible contribution of Trichoderma VOCs in antagonistic processes against grapevine downy mildew has not yet been investigated. In this work, VOC emission profiles of three Trichoderma strains belonging to T. asperellum, T. atroviride and T. harzianum were analysed using headspace-solidphase microextraction gas chromatography-mass spectrometry (HS-SPME/GC-MS). Total ion current chromatograms were processed by an open source software [3], and statistical analysis on the obtained raw data was carried out using an in-house R-script. VOCs emitted by the Trichoderma strains at specific time-points were selected, and pure compounds were tested against downy mildew by leaf disks assays on susceptible grapevines. The development of downy mildew symptoms was reduced on leaf disks exposed to air treated with two sesquiterpenes, one hydrocarbon, or one heterocyclic compound, indicating the efficacy of these VOCs against downy mildew in plant tissues. Our data suggest that VOCs emitted by the Trichoderma strains under study can effectively be detected and identified by HS-SPME/GC-MS, and can inhibit the development of downy mildew symptoms on susceptible grapevine
Simulated global warming affects endophytic bacterial and fungal communities of Antarctic pearlwort leaves and some bacterial isolates support plant growth at low temperatures
No full textAntarctica is one of the most stressful environments for plant life and the Antarctic pearlwort (Colobanthus quitensis) is adapted to the hostile conditions. Plant-associated microorganisms can contribute to plant survival in cold environments, but scarce information is available on the taxonomic structure and functional roles of C. quitensis-associated microbial communities. This study aimed at evaluating the possible impacts of climate warming on the taxonomic structure of C. quitensis endophytes and at investigating the contribution of culturable bacterial endophytes to plant growth at low temperatures. The culture-independent analysis revealed changes in the taxonomic structure of bacterial and fungal communities according to plant growth conditions, such as the collection site and the presence of open-top chambers (OTCs), which can simulate global warming. Plants grown inside OTCs showed lower microbial richness and higher relative abundances of biomarker bacterial genera (Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Aeromicrobium, Aureimonas, Hymenobacter, Novosphingobium, Pedobacter, Pseudomonas and Sphingomonas) and fungal genera (Alternaria, Cistella, and Vishniacozyma) compared to plants collected from open areas (OA), as a possible response to global warming simulated by OTCs. Culturable psychrotolerant bacteria of C. quitensis were able to endophytically colonize tomato seedlings and promote shoot growth at low temperatures, suggesting their potential contribution to plant tolerance to cold conditions
Effects of blood flow restriction on spine postural control using a robotic platform: A pilot randomized cross-over study
No full textBACKGROUND: Blood flow restriction (BFR) training improves muscle strength and functional outcomes, but the proprioceptive implications of this technique in the rehabilitation field are still unknown. OBJECTIVE: The present study aimed at assessing the effects of BFR in terms of stabilometric and balance performance. METHODS: In this pilot randomized cross-over study, healthy young adults were included and randomly assigned to Groups A and B. Both groups underwent a postural assessment with and without wearing a BFR device. Study participants of Group A underwent postural baseline assessment wearing BFR and then removed BFR for further evaluations, whereas subjects in Group B performed the baseline assessment without BFR and then with BFR. Stabilometric and balance performance were assessed by the robotic platform Hunova, the Balance Error Scoring System (BESS), the self-reported perceived balance (7-point Likert scale), and discomfort self-rated assessment. Moreover, the safety profile was recorded. RESULTS: Fourteen subjects were included and randomly assigned to Group A (n: 7) and Group B (n: 7). Significant differences were shown in balance tests in static conditions performed on the Hunova robot platform in terms of average distance RMS (root-mean-square) with open eyes (OE), anteroposterior (AP) trunk oscillation range with OE, mediolateral (ML) average speed of oscillation with OE, and total excursion AP range with closed eyes (CE) (BFR: 3.44 ± 1.06; without BFR: 2.75 ± 0.72; p= 0.041). Moreover, elastic balance test showed differences in Romberg index (BFR: 0.16 ±0.16; without BFR: 0.09 ± 0.07; p= 0.047). No adverse events were reported. CONCLUSION: Taken together, our data showed that BFR affects balance performance of healthy subjects. Further studies are needed to better characterize the possible role of BFR treatment in the context of a specific rehabilitation protocol
Identification of Trichoderma spp. volatile organic compounds (VOCs) by HS-SPME/GC-MS analysis
No full textIntroduction
Trichoderma spp. are among the most widespread soil microorganisms and have been widely used as biological control agents against numerous phytopathogens [1]. Trichoderma biological control mechanisms are based on the production of antimicrobial substances, lytic enzymes, competition with other microorganisms for nutrients and/or space, mycoparasitism, and plant resistance induction [2]. Moreover, Trichoderma spp. are known to produce numerous volatile organic compounds (VOCs) [3] that play crucial roles in the inter-kingdom communications and biological control mechanisms [4]. Three Trichoderma strains, such as T. asperellum T34 (T34), T. atroviride SC1 (SC1), T. harzianum T39 (T39) are well documented for their ability to cope with a broad spectrum of phytopathogens and are commercialised as biofungicides. Several studies highlighted the importance of Trichoderma spp. as an alternative strategy against grapevine pathogens [5], but no information is available on the possible biological control mechanism mediated by Trichoderma VOCs against grapevine downy mildew (caused by Plasmopara viticola). The aim of this study was to identify VOCs produced by T34, SC1 and T39 using headspace-solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME/GC-MS) analysis.
Materials and Methods
Volatile-mediated effects of Trichoderma strains were assessed on grapevine leaf disks incubated with uninoculated potato dextrose agar (PDA) Petri dishes (Control) and dishes with PDA-grown T34, SC1 or T39 colonies without contact with leaf tissues. Leaf disks were inoculated with a suspension of P. viticola sporangia and downy mildew severity was assessed at six days post inoculation (dpi).
For headspace analysis of Trichoderma spp. VOCs, 20 μL of the conidial suspension of the respective Trichoderma strain were inoculated on PDA poured into sterile 20 mL-HS vials. VOCs produced by Trichoderma strains were measured using HS-SPME/GC-MS analysis at 48 or 72 h of incubation at 25°C. For compound identification, deconvoluted mass spectra were compared with an in-house library obtained with authentic reference standards. Compound annotation was achieved imposing a relative deviation of retention index (RI) value lower than 2% from the reference value published in the NIST14 database, and according to the mass spectrum similarity score set higher than 70%. Two independent experiments were carried out. Five of the identified VOCs were selected according to their emission profiles and pure compounds were tested against P. viticola by leaf disk assay.
Results
VOCs produced by T34, SC1 or T39 reduced downy mildew symptoms on grapevine leaf disks. HS-SPME/GC-MS analysis revealed a total of 26 and 21 Trichoderma VOCs found in the first and second experiments, respectively. Trichoderma VOCs included alkenes, ketones, pyrones, furanes and terpenes. Terpenes were successfully identified using manual inspection of extracted ion current (EIC) chromatograms at m/z 136 for monoterpenes, m/z 202 and 204 for sesquiterpenes, and m/z 272 for diterpenes. VOC profiles differed according to the Trichoderma strains and T39 was the highest producer of VOCs and terpenes compared to T34 and SC1. Five metabolite groups were identified according to changes in abundance among the three Trichoderma strains: VOCs with consistent changes in both experiments at both (Group 1), or at one time point (Group 2); VOCs with different changes in abundance in both experiments (Group 3); VOCs detected only in the first (Group 4) or second (Group 5) experiment. Five VOCs (two sesquiterpenes, one alkene, one furan and one pyrone) were tested against P. viticola and each VOC was applied to a filter paper disk on the lid of the Petri dish without contact with leaf tissues. Three VOCs slightly reduced the development of downy mildew symptoms at dosages of 50 mg/L in air volume. Two pure VOCs significantly reduced downy mildew symptoms at the dosage of 10 mg/L in air volume.
Discussion/Conclusions
HS-SPME/GC-MS analysis is a powerful tool to detect and identify Trichoderma VOCs. VOC profiles differed in the three Trichoderma strains and T39 was the highest producer of terpenes, followed by T34 and SC1, indicating genotypic determinants of VOC production. Five VOCs (two sesquiterpenes, one alkene, one furan and one pyrone) reduced downy mildew symptoms on grapevine leaf disks. Preliminary results suggest that two most effective VOCs enhanced the grapevine defence responses against P. viticola. These results indicate that Trichoderma VOCs are promising molecules that could be further developed to naturally control grapevine downy mildew. Further transcriptomic and metabolomic studies are required to shed light on the grapevine defence mechanisms activated by Trichoderma VOCs against downy mildew
Avaliação pré-clínica do potencial antiinflamatório e cicatrizante da Malva Sylvestris Linn. em alterações bucais
No full textOrientadora : Michel OtukiCo-orientadora : Daniela de Almeida CabriniDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia. Defesa: Curitiba, 27/03/2008Bibliografia f.105-123Resumo: A inflamação é uma resposta de proteção do organismo a uma agente agressor, cujo objetivo é limitar ou destruir esse agente. O periodonto, como parte integrante do organismo está sujeito a sofrer alterações inflamatórias. Ele é a unidade formada pela gengiva, osso alveolar, ligamento periodontal e cemento radicular. A partir da susceptibilidade do indivíduo, seguindo-se de acúmulo de placa bacteriana, as alterações inflamatórias se manifestam na forma inicial de gengivite, podendo ou não evoluir para uma periodontite. A doença periodontal é de difícil tratamento, por se tratar de vários microrganismos envolvidos, em sua etiologia. Além dos diversos agentes quimioterápicos, estudos têm sido realizados para se verificar a ação de determinados produtos naturais. Uma das plantas que, de certa forma, é muito recomendada e utilizada por indivíduos com problemas bucais, mas de forma empírica, é a Malva sylvestris L. A partir dos resultados obtidos sobre a ação antiinflamatória da M. sylvestris, em doença periodontal induzida por ligadura, tornou-se de interesse para o trabalho avaliar também o processo de cicatrização da planta em lesão palatina de ratos. Os resultados obtidos nos mostraram que a M. sylvestris, na dose de 25 mg/dente reduziu a atividade das enzimas mieloperoxidase (MPO) e NAG, também houve redução da reabsorção óssea e aumento do percentual ósseo periodontal. A planta também se apresentou eficaz, in vitro, na redução da liberação de óxido nítrico (NO) por fibroblastos gengivais estimulados por lipopolissacarídeo (LPS). Na dose de 50 mg/lesão reduziu a atividade das enzimas mieloperoxidase (MPO) e NAG, e também o tempo de cicatrização. A partir desses resultados sugere-se que a M. sylvestris, pela ação sinérgica de seus compostos, possui ação antiinflamatória e cicatrizante no tratamento da doença periodontal induzida por ligadura e na cicatrização de lesão palatina, necessitando ainda de mais estudos.Abstract: Inflammation is a protection response of organism, designed to eradicate irritant agents. The periodontium, part of the masticatory system, as any part of the body is subject to change inflammatory. It is defined as those tissues supporting and investing the tooth, comprises root cementum, periodontal ligament, alveolar bone and gingival. From the susceptibility of the individual, followed by the accumulation of plaque, the periodontium can change inflammatory, which is manifested in the initial form of gingivitis, which may or not evolves into a periodontitis. The periodontal disease is difficult to treat, because it is different microorganisms involved in its etiology. In addition to various chemotherapeutic agents such as antibiotics, nonsteroidal antiinflammatory drugs, anti-septic, metal ions and agents oxigenants, studies have been conducted to verify the action of certain natural products. One of the plants that, in a way, it is very recommended and used by individuals with dental problems, but so empirical, is the Malva sylvestris L.. From the results obtained on the action of M. sylvestris, in periodontal disease induced by ligature, it became of interest to the work also evaluate the healing process of the plant in palate injury of rats. The results showed that M. sylvestris, the dose of 25 mg/tooth reduced the activity of enzymes myeloperoxidase (MPO) and NAG, there was also reduction of bone and increasing the percentage periodontal bone. The plant also made effective in vitro, in reducing the release of nitric oxide (NO) by gingival fibroblasts stimulated by lipopolissacarídeo (LPS). At the dose of 50 mg/injury reduced the activity of enzymes myeloperoxidase (MPO) and NAG, and also the time for healing. From these results suggest that the M. sylvestris, through the synergistic action of its compounds, has anti-inflammatory action and healing in the treatment of periodontal
disease induced by ligature and the healing palate, requiring even more studies
Structural properties of silica gels prepared from oxalic-acid-catalyzed tetraethoxysilane sonohydrolysis
No full textAbout similar to 2.1 x 10(-3) Mol SiO2 cm(-3) and similar to 88%-volume liquid-phase silica wet gels were prepared from oxalic-acid-catalyzed tetraethoxysilane (TEOS) sonohydrolysis. Aerogels were obtained by supercritical CO2 extraction. The samples were analyzed by thermogravimetry, small-angle X-ray scattering and nitrogen adsorption. Wet gels can be described as mass fractal structures with fractal dimension D similar to 1.94 and structural characteristic length zeta changing between similar to 3.3 to similar to 3.0 nm in the studied range of the catalyst concentration. A fraction of the porosity is apparently eliminated in the supercritical process. The values of the BET specific surface S-BET, the total pore volume V-p and the mean pore size l(p) of the aerogels were found to change almost randomly around the mean values S-BET = 874 m(2) g(-1), V-p = 0.961 cm(3) g(-1) and l(p) = 4.4 nm with catalyst concentration variation. These values were not substantially different from those from an equivalent HCl-catalyzed aerogel. (c) 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.Univ Estadual Paulista, UNESP, IGCE, Dept Fis, BR-13500970 Rio Claro, SP, BrazilUniv Estadual Paulista, UNESP, IGCE, Dept Fis, BR-13500970 Rio Claro, SP, Brazi
Breast Histologic Changes In Female Rats Treated With Sex Steroids [histolologia Mamária Após Uso De Esteróides Sexuais - Estudo Em Ratas]
No full textOBJECTIVE. To evaluate the association of histologic changes in the breasts of female rats undergoing therapy with sex steroids. METHODS. An experimental study was conducted of 40 castrated female non-pubertal rats, 20 had given birth (Group B) and 20 had no offspring (Group A). After four weeks, these rats were randomly allocated to subgroups: A1, A2, A3, A4, A5 and B1, B2, B3, B4, B5. Steroids were given to subgroups as follows: A1 and B1 - estradiol benzoate; A2 and B2 - medroxyprogesterone acetate; A3 and B3 - estradiol benzoate and medroxyprogesterone acetate; A4 and B4 - tibolone; A5 and B5 - placebo. After 10 weeks of treatment, animals were sacrificed and their mammary glands were analyzed. Histologic parameters evaluated were: epithelial cell proliferation, epithelial cells with secretory activity; and cell atypia in terminal duct units and buds or terminal alveoli. The association between microscopic analysis and diverse therapeutic regimens were analyzed by calculating the odds ratio and its respective 95% confidence interval. RESULTS. Histologic changes were observed in 29 rats: moderate hyperplasia (52.5%), hyperplastic alveolar nodule (42.5%), epithelial atypia (35%), mild hyperplasia (32.5%), secretory activity (20%) and severe hyperplasia (5%). In rats with no offspring when compared to the control, 1.3 times more hyperplastic alveolar nodules were found in the group treated with estradiol, the same was true for moderate hyperplasia in the rats that received medroxyprogesterone acetate, hyperplastic alveolar nodules and epithelial atypia in the group treated with estradiol plus medroxyprogesterone acetate. In the rats with offspring 1.3 times more secretory activity was found with estradiol. CONCLUSION. 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