Molecular Mechanisms Underlying Sugarcane Response to Aluminum Stress by RNA-Seq.

Some metals are beneficial to plants and contribute to critical physiological processes. Some metals, however, are not. The presence of aluminum ions (Al3+) can be very toxic, especially in acidic soils. Considerable parts of the world's arable land are acidic in nature; mechanistically elucidating a plant's response to aluminum stress is critical to mitigating this stress and improving the quality of plants. To identify the genes involved in sugarcane response to aluminum stress, we generated 372 million paired-end RNA sequencing reads from the roots of CTC-2 and RB855453, which are two contrasting cultivars. Data normalization resulted in 162,161 contigs (contiguous sequences) and 97,335 genes from a de novo transcriptome assembly (trinity genes). A total of 4858 and 1307 differently expressed genes (DEGs) for treatment versus control were identified for the CTC-2 and RB855453 cultivars, respectively. The DEGs were annotated into 34 functional categories. The majority of the genes were upregulated in the CTC-2 (tolerant cultivar) and downregulated in RB855453 (sensitive cultivar). Here, we present the first root transcriptome of sugarcane under aluminum stress. The results and conclusions of this study are a crucial launch pad for future genetic and genomic studies of sugarcane. The transcriptome analysis shows that sugarcane tolerance to aluminum may be explained by an efficient detoxification mechanism combined with lateral root formation and activation of redox enzymes. We also present a hypothetical model for aluminum tolerance in the CTC-2 cultivar.

Essential oils of Citrus aurantifolia, Anthemis nobile and Lavandula officinalis: in vitro anthelmintic activities against Haemonchus contortus.

BACKGROUND: Infections of sheep with gastrointestinal parasites, especially Haemonchus contortus, have caused serious losses in livestock production, particularly after the emergence of resistance to conventional anthelmintics. The search for new anthelmintic agents, especially those of botanical origin, has grown substantially due to the perspective of less contamination of meat and milk, as well as other advantages related to their cost and accessibility in less developed countries. The aim of this study was to evaluate the in vitro anthelmintic activity of essential oils of the plant species Citrus aurantifolia, Anthemis nobile and Lavandula officinalis against the main developmental stages of the parasite H. contortus. RESULTS: Plant species were selected based on substantial ethnopharmacological information. Analysis of the composition of each oil by gas chromatography coupled to mass spectrometry (GC-MS) demonstrated the presence of limonene (56.37%), isobutyl angelate (29.26%) and linalool acetate (35.97%) as the major constituents in C. aurantifolia, A. nobile and L. officinalis, respectively. Different concentrations of each oil were tested in vitro for their capacity to inhibit egg hatching (EHT), larval development (LDT) and adult worm motility (AWMT) using a multidrug-resistant strain of H. contortus (Embrapa 2010). The IC50 values obtained for the oils of C. aurantifolia, A. nobile and L. officinalis were 0.694, 0.842 and 0.316 mg/ml in the EHT and 0.044, 0.117 and 0.280 mg/ml in the LDT, respectively. The three oils were able to inhibit adult worm motility completely within the first 8-12 h of observation in the AWMT. CONCLUSIONS: The present results demonstrate significant anthelmintic activity of the three oils against the different developmental stages of H. contortus. Furthermore, this study is of ethnopharmacological importance by validating the anthelmintic activity of the oils studied. Although new experiments are necessary, these data contribute to the development of pharmaceutical-veterinary products for sheep farming by opening up new therapeutic possibilities against gastrointestinal infections caused by H. contortus.

Aqueous extracts from Uncaria tomentosa (Willd. ex Schult.) DC. reduce bronchial hyperresponsiveness and inflammation in a murine model of asthma.

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria tomentosa (Willd. Ex Schult) DC is used by indigenous tribes in the Amazonian region of Central and South America to treat inflammation, allergies and asthma. The therapeutic properties of U. tomentosa have been attributed to the presence of tetracyclic and pentacyclic oxindole alkaloids and to phenolic acids. AIMS OF THE STUDY: To characterize aqueous bark extracts (ABE) and aqueous leaf extracts (ALE) of U. tomentosa and to compare their anti-inflammatory effects. MATERIALS AND METHODS: Constituents of the extracts were identified by ultra performance liquid chromatography-mass spectrometry. Anti-inflammatory activities were assessed in vitro by exposing lipopolysaccharide-stimulated macrophage cells (RAW264.7-Luc) to ABE, ALE and standard mitraphylline. In vivo assays were performed using a murine model of ovalbumin (OVA)-induced asthma. OVA-sensitized animals were treated with ABE or ALE while controls received dexamethasone or saline solution. Bronchial hyperresponsiveness, production of Th1 and Th2 cytokines, total and differential counts of inflammatory cells in the bronchoalveolar lavage (BAL) and lung tissue were determined. RESULTS: Mitraphylline, isomitraphylline, chlorogenic acid and quinic acid were detected in both extracts, while isorhyncophylline and rutin were detected only in ALE. ABE, ALE and mitraphylline inhibited the transcription of nuclear factor kappa-B in cell cultures, ALE and mitraphylline reduced the production of interleukin (IL)-6, and mitraphylline reduced production of tumor necrosis factor-alpha. Treatment with ABE and ALE at 50 and 200 mg kg-1, respectively, reduced respiratory elastance and tissue damping and elastance. ABE and ALE reduced the number of eosinophils in BAL, while ALE at 200 mg kg-1 reduced the levels of IL-4 and IL-5 in the lung homogenate. Peribronchial inflammation was significantly reduced by treatment with ABE and ALE at 50 and 100 mg kg-1 respectively. CONCLUSION: The results clarify for the first time the anti-inflammatory activity of U. tomentosa in a murine model of asthma. Although ABE and ALE exhibited distinct chemical compositions, both extracts inhibited the production of pro-inflammatory cytokines in vitro. In vivo assays revealed that ABE was more effective in treating asthmatic inflammation while ALE was more successful in controlling respiratory mechanics. Both extracts may have promising applications in the phytotherapy of allergic asthma.

Phototherapy (LEDs 660/890nm) in the treatment of leg ulcers in diabetic patients: case study.

This study evaluated the use of phototherapy in the healing of mixed leg ulcers in two diabetic patients (type 2) with arterial hypertension. The device had probe 1 (one 660 nm LED, 5 mW) applied in 3 ulcers and probe 2 (32 890 nm LEDs associated with 4 660 nm LEDs, 500 mW) in 6 ulcers. After asepsis, ulcers were treated with probes to 3 J/cm2, 30 sec per point, twice a week, followed by topical daily dressing with 1% silver sulphadiazine during 12 weeks. The following analyses of ulcers with software Image J showed that probe 2 presented mean healing rates of 0.6; 0.7 and 0.9, whereas probe 1 had 0.2; 0.4 and 0.6 at 30, 60 and 90 days, respectively. Phototherapy accelerated wound healing of leg ulcers in diabetic patients.

Morphologic and morphometric analysis of the root canal apical third cleaning after biomechanical preparation using 3.3% Ricinus communis detergent and 1% NaOCl as irrigating solutions.

This study evaluated, by morphologic and morphometric analyses, the cleaning of apical third of root canals instrumented with nickel-titanium rotary files using different irrigating solutions. Twenty-seven single-rooted mandibular premolars were assigned to three groups (n=9), according to the irrigating solution used: Group I, distilled and deionized water; Group II, 1% NaOCl; and Group III, 3.3% Ricinus communis detergent. Biomechanical preparation was performed with Protaper Plusâ nickel-titanium files as follows: S1, SX and S2 at the cervical and middle thirds, and 25/02, 25/04, 25/06, 30/02, 30/04 and 30/06 to complete the instrumentation, operating at 1 mm from the root apex. Irrigation was done at each file change with 2 mL of irrigating solution, totalizing 20 mL for each tooth. After biomechanical preparation, the apical thirds were serially sectioned and histologically processed. The cross-sections were examined by an optical microscope (X40) connected to a computer. The images were captured and analyzed using a computer software and submitted to morphometric analysis with aid of a grid. The percentage of debris remaining at the apical third was calculated. Data were submitted to statistical analysis by ANOVA and Tukey test. The results showed no statistically significant difference (p>0.01) between the groups irrigated with 1% NaOCl and 3.3% Ricinus communis detergent, which presented the lowest percentage of debris at the apical third, 8.49% and 10.11%, respectively. The group irrigated with distilled and deionized water had the highest percentage of debris (15.58%) and was statistically different from the other groups (p<0.01). It may be concluded that 3.3% Ricinus communis detergent and 1% NaOCl had similar cleaning effectiveness on removal of debris from root canals.

Fototerapia (LEDs 660/890nm) no tratamento de úlceras de perna em pacientes diabéticos: estudo de caso / Phototherapy (LEDs 660/890nm) in the treatment of leg ulcers in diabetic patients: case study

Avaliou-se a fototerapia na cicatrização de úlceras de perna (UP) mistas em dois pacientes diabéticos (tipo 2), hipertensos. O aparelho apresentava sonda 1 (S1) (1 LED de 660nm, 5mW) aplicado em 3 UP e sonda 2 (S2) (32 LEDs de 890nm e 4 LEDs de 660nm, 500mW) em 6 UP. Após antissepsia,úlceras foram tratadas com sondas a 3J/cm2, 30seg, 2x/semana seguido pelo curativo diário com sulfadiazina de prata a 1 por cento por 12 semanas. Pela análise com software Image J®, as UP com S2 tiveram índices de cicatrização médios de 0,6; 0,7 e 0,9 enquanto S1 foi de 0,2; 0,4 e 0,6 no 30º, 60º e 90º dias, respectivamente. A fototerapia acelerou a cicatrização das úlceras de perna em pacientes diabéticos.

This study evaluated the use of phototherapy in the healing of mixed leg ulcers in two diabetic patients (type 2) with arterial hypertension. The device had probe 1 (one 660nm LED, 5mW) applied in 3 ulcers and probe 2 (32 890nm LEDs associated with 4 660nm LEDs, 500mW) in 6 ulcers. After asepsis, ulcers were treated with probes to 3 J/cm2, 30sec per point, twice a week, followed by topical daily dressing with 1 percent silver sulphadiazine during 12 weeks. The following analyses of ulcers with software Image J showed that probe 2 presented mean healing rates of 0.6; 0.7 and 0.9, whereas probe 1 had 0.2;0.4 and 0.6 at 30, 60 and 90 days, espectively. Phototherapy accelerated wound healing of leg ulcers in diabetic patients.

Quantitative determination for cytotoxic Friedo-nor-oleanane derivatives from five morphological types of Maytenus ilicifolia (Celastraceae) by reverse-phase high-performance liquid chromatography.

Five different morphological types of Maytenus ilicifolia of the same age and harvested under the same conditions showed distinct accumulations of some friedo-nor-oleananes. A rapid, sensitive and reliable reverse-phase HPLC method (employing an external standard) was used for the determination of the cytotoxic triterpenoids, 20 alpha-hydroxymaytenin, 22 beta-hydroxymaytenin, maytenin, celastrol and pristimerin in each of the five types. Well resolved peaks with good detection response and linearity in the range 1.0-100 micrograms/mL were obtained.

Antioxidant flavan-3-ols and flavonol glycosides from Maytenus aquifolium.

TLC autographic assay revealed, in the EtOAc extract obtained from leaves and root bark of Maytenus aquifolium (Celastraceae), the presence of fi ve compounds exhibiting antioxidant properties towards beta-carotene. They were isolated and identified as epigallocatechin (1), (+) ouratea-catechin (2), proanthocyanidin (3), kaempferol 3-O-alpha-L-rhamnopyranosyl (1-->6)-O-[beta-D-glucopyranosyl (1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)]-O-beta-D-glucopyranosyl (4) and quercetin 3-O-alpha-L-rhamnopyranosyl (1-->6)-O-[beta-D-glucopyranosyl (1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)]-O-beta-D-glucopyranosyl (5). The isolates were investigated for their redox properties using cyclic voltammetry and for their radical scavenging abilities through spectrophotometric assay on the reduction of 2,2-diphenyl-pycryl hydrazyl (DPPH). These results were correlated to the inhibition of beta-carotene bleaching on TLC autographic assay and to structural features of the flavonoids.

Quantitative determination of anti-fungal and insecticide amides in adult plants, plantlets and callus from Piper tuberculatum by reverse-phase high-performance liquid chromatography.

A rapid, sensitive and reliable reverse-phase HPLC method was used for the quantitative determination of the anti-fungal and insecticide amides, dihydropiplartine (1), piplartine (2), deltaalpha,beta-dihydropiperine (3) and pellitorine (4) in plants in natura, in plantlets in vitro and ex vitro, and in callus of Piper tuberculatum. Well-resolved peaks were obtained with good detection response and linearity in the range of 15.0-3000 microg/mL. The plants in natura contained compounds 1-4, the plantlets ex vitro and in vitro accumulated compounds 1-2 and 1-4, respectively, while only amide 4 was found in callus.

Production of an antiproliferative furanoheliangolide by Lychnophora ericoides cell culture.

This work reports for the first time the production a furanoheliangolide (goyazensolide) by plant cell culture. Monitoring of the goyazensolide metabolism revealed that the maximum production occurred during the lag phase of the Lychnophora ericoides callus culture. The antiproliferative activity of obtained goyazensolide was evaluated against seven cancer cell lines using MTT assay. The results revealed a potent cytotoxic activity for the furaheliangolide with IC50 values in the range of 0.06 microg/ml for CEM leukemia cells to 0.75 microg/ml for B16 melanome cells.

Morphologic and morphometric analysis of the root canal apical third cleaning after biomechanical preparation using 3.3 percent Ricinus communis detergent and 1 percent NaOCl as irrigating solutions%Análise morfológica e morfométrica da limpeza de canais radiculares após preparo biomecânico utilizando detergente derivado do óleo da mamona (Ricinus communis) a 3.3% e NaOCl a 1% como soluções irrigantes

Estudou-se, por meio de análise morfológica e morfométrica, a capacidade de limpeza promovida pela instrumentação rotatória com limas Ni-Ti e irrigação com diferentes soluções. Vinte e sete pré-molares inferiores foram distribuídos em três grupos (n=9), de acordo com a solução irrigante utilizada: Grupo I, água destilada e deionizada; Grupo II, NaOCl a 1% e Grupo III, detergente derivado do óleo de mamona (Ricinus communis) a 3,3%. O preparo biomecânico foi realizado com sistema rotatório de Ni-Ti Protaper Plus, obedecendo a seguinte ordem de uso dos instrumentos: S1, SX e S2 para o terço cervical e médio, seguida do 25/02, 25/04, 25/06, 30/02, 30/04 e 35/02, completando a instrumentação todos operando 1 mm aquém do ápice. A irrigação foi realizada a cada troca de instrumento com 2 mL de solução irrigante, totalizando um volume de 20 mL para cada dente. Após o preparo biomecânico, os terços apicais dos dentes foram submetidos ao processamento histológico. Os espécimes foram analisados em microscópio óptico (40X) conectado a um computador. As imagens foram capturadas e analisadas utilizando-se softwares específicos e submetidas à análise morfométrica por meio de uma grade de integração. A porcentagem de debris presente no terço apical dos canais foi calculada. Os dados foram submetidos à análise estatística por meio de ANOVA e teste de Tukey. Os resultados do presente estudo evidenciaram que não houve diferença estatisticamente significante (p>0,01) entre os grupos irrigados com NaOCl a 1% e detergente de mamona a 3,3%, que apresentaram menor porcentagem de debris no terço apical, 8,49% e 10,11% respectivamente. O grupo irrigado com água destilada e deionizada apresentou a maior porcentagem de debris (15,58%), sendo estatisticamente diferente (p<0,01) dos outros grupos experimentais. Concluiu-se que o detergente de mamona a 3,3% apresentou efetividade semelhante ao NaOCl a 1% na remoção de debris dos canais radiculares

Morphologic and morphometric analysis of the root canal apical third cleaning after biomechanical preparation using 3.3 percent Ricinus communis detergent and 1 percent NaOCl as irrigating solutions

This study evaluated, by morphologic and morphometric analyses, the cleaning of apical third of root canals instrumented with nickel-titanium rotary files using different irrigating solutions. Twenty-seven single-rooted mandibular premolars were assigned to three groups (n=9), according to the irrigating solution used: Group I, distilled and deionized water; Group II, 1 percent NaOCl; and Group III, 3.3 percent Ricinus communis detergent. Biomechanical preparation was performed with Protaper Plusâ nickel-titanium files as follows: S1, SX and S2 at the cervical and middle thirds, and 25/02, 25/04, 25/06, 30/02, 30/04 and 30/06 to complete the instrumentation, operating at 1 mm from the root apex. Irrigation was done at each file change with 2 mL of irrigating solution, totalizing 20 mL for each tooth. After biomechanical preparation, the apical thirds were serially sectioned and histologically processed. The cross-sections were examined by an optical microscope (X40) connected to a computer. The images were captured and analyzed using a computer software and submitted to morphometric analysis with aid of a grid. The percentage of debris remaining at the apical third was calculated. Data were submitted to statistical analysis by ANOVA and Tukey test. The results showed no statistically significant difference (p>0.01) between the groups irrigated with 1 percent NaOCl and 3.3 percent Ricinus communis detergent, which presented the lowest percentage of debris at the apical third, 8.49 percent and 10.11 percent, respectively. The group irrigated with distilled and deionized water had the highest percentage of debris (15.58 percent) and was statistically different from the other groups (p<0.01). It may be concluded that 3.3 percent Ricinus communis detergent and 1 percent NaOCl had similar cleaning effectiveness on removal of debris from root canals.

Estudou-se, por meio de análise morfológica e morfométrica, a capacidade de limpeza promovida pela instrumentação rotatória com limas Ni-Ti e irrigação com diferentes soluções. Vinte e sete pré-molares inferiores foram distribuídos em três grupos (n=9), de acordo com a solução irrigante utilizada: Grupo I, água destilada e deionizada; Grupo II, NaOCl a 1 por cento e Grupo III, detergente derivado do óleo de mamona (Ricinus communis) a 3,3 por cento. O preparo biomecânico foi realizado com sistema rotatório de Ni-Ti Protaper Plus, obedecendo a seguinte ordem de uso dos instrumentos: S1, SX e S2 para o terço cervical e médio, seguida do 25/02, 25/04, 25/06, 30/02, 30/04 e 35/02, completando a instrumentação todos operando 1 mm aquém do ápice. A irrigação foi realizada a cada troca de instrumento com 2 mL de solução irrigante, totalizando um volume de 20 mL para cada dente. Após o preparo biomecânico, os terços apicais dos dentes foram submetidos ao processamento histológico. Os espécimes foram analisados em microscópio óptico (40X) conectado a um computador. As imagens foram capturadas e analisadas utilizando-se softwares específicos e submetidas à análise morfométrica por meio de uma grade de integração. A porcentagem de debris presente no terço apical dos canais foi calculada. Os dados foram submetidos à análise estatística por meio de ANOVA e teste de Tukey. Os resultados do presente estudo evidenciaram que não houve diferença estatisticamente significante (p>0,01) entre os grupos irrigados com NaOCl a 1 por cento e detergente de mamona a 3,3 por cento, que apresentaram menor porcentagem de debris no terço apical, 8,49 por cento e 10,11 por cento respectivamente. O grupo irrigado com água destilada e deionizada apresentou a maior porcentagem de debris (15,58 por cento), sendo estatisticamente diferente (p<0,01) dos outros grupos experimentais. Concluiu-se que o detergente de mamona a 3,3 por cento apresentou efetividade semelhante ao...