Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract / Supressão de Xanthomonas oryzae pv. oryzae: formação do biofilme oryzae pelo extrato de folhas de metanol de Acacia mangium

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract.

Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Effect of fixation method on intergonial width stability after mandibular setback via bilateral sagittal split osteotomy.

Q4: Horizontal changes occur following bilateral sagittal split osteotomy (BSSO) in skeletal class III patients. The aim of this study was to assess the ostoperative changes in intergonial (IG) width and compare them between the positional screw and miniplate fixation methods in BSSO. This study evaluated patients who had mandibular prognathism and underwent BSSO for mandibular setback. Internal fixation was performed bilaterally, either with positional screws in the lateral ramus or with a miniplate. Postero-anterior cephalograms were obtained preoperatively (T1), at 1 month postoperative (T2), and at 6 months postoperative (T3). The IG widths and the alterations in IG width postoperative (T2-T1, T3-T2) were measured. No correlations were observed between the amount of setback and changes at T2 -T1 or T3-T2. The IG width values decreased after mandibular setback and internal fixation with both methods. Statistical analyses showed a significant difference between T3 and T1 in the miniplate group (P=0.045). No significant difference in the postoperative change in IG width (T2-T1 and T3-T2) was found between the two fixation groups. The magnitude of this change was smaller for positional screws when compared to miniplates for fixation. The amount of mandibular setback showed no correlation with postoperative changes in IG width..

Luliconazole, a new antifungal against Candida species isolated from different sources.

OBJECTIVE: Luliconazole is an inhibitor for sterol 14-α-demethylase in fungal cells with a broad-spectrum antifungal activity against dermatophytes, Candida albicans, Malassezia species, dematiaceous and hyaline hyphomycetes. Furthermore, luliconazole has been clinically used for the treatment of pityriasis versicolor, dermatophytosis, onychomycosis, cutaneous and mucocutaneous candidiasis. In the present study, we aimed to evaluate in vitro antifungal activity of luliconazole against several strains of Candida species recovered from different clinical materials. MATERIALS AND METHODS: In the present study, 104 strains of Candida species including, 34 isolates from vaginitis, 23 isolates from AIDS patients with vaginal candidiasis, 24 isolates from neutropenic patients and 24 isolates from tracheal tubes, were examined for susceptibility tests. A serial dilution of luliconazole (4-0.008µg/mL) was tested against different strains of Candida species recovered from different sources. RESULTS: The minimum inhibitory concentration (MIC) range and MIC90 of vaginal isolates (HIV-) were 1-0.063 and 1µg/mL. Furthermore, the most of strains (50%) had a MIC of 0.5µg/mL. The MIC ranges were similar (2-0.016µg/mL) for both vaginal (HIV+) and neutropenic patients isolates, whereas, MIC90 for them were 0.5 and 1µg/mL, respectively. All tracheal tubes strains were inhibited at the range of 2-0.008µg/mL with MIC90=1µg/mL. Totally, the lowest MIC50 (MIC=0.015µg/mL), MIC90 (MIC=1µg/mL) and MICGM (MIC=0.05µg/mL) are correlated to C. glabrata, a non-albicans species. CONCLUSION: It is concluded that, luliconazole could be an alternative anti-Candida agent, however, in vivo studies must be confirmed usefulness of drug for clinical usage.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Effect of extracellular matrix on bovine spermatogonial stem cells and gene expression of niche factors regulating their development in vitro.

Extracellular matrix (ECM) could influence cells function through providing structural and functional networks facilitating the cellular interactions. The present study was conducted to evaluate the effect of culture on ECM versus plastic on bovine spermatogonial stem cells (SSCs) and growth factors regulating their development. Following isolation, bovine testicular cells were cultured on ECM-coated or uncoated (control) plates for 12 days. The colonization of SSCs was assessed by inverted microscope and the gene expression was evaluated using quantitative real-time PCR. The colonization rate was greater in ECM than the control group (P<0.05). The expression of markers of undifferentiated spermatogonia increased in response to conventional culture (P<0.05). Conversely, the expression of ckit as a marker for differentiated spermatogonia was reduced following culture in the control and ECM groups (P<0.05), but this decrease was less in ECM group (P<0.05). Accordingly, while cells cultured on uncoated plates had greater expression of markers of undifferentiated spermatogonia (P<0.05), cells cultured on ECM-coated plates showed higher expression of ckit (P<0.05). Moreover, culture on ECM resulted in higher expression of kit ligand (Kitlg; P<0.05), whereas culture on plastic led to greater expression of glial cell line-derived neurotrophic factor (Gdnf; P<0.05). In conclusion, the present study revealed that the permissive effect of ECM on bovine SSCs differentiation in vitro, which was probably mediated through upregulation of KITLG expression. Moreover, the results imply that GDNF might contribute to germ cells self-renewal during conventional culture.

Investigation of EEG changes during exposure to extremely low-frequency magnetic field to conduct brain signals.

There are evidences that confirm the effect of magnetic fields (MFs) on brain signals and some psychological disorders such as headache, migraine and depression. The aim of the present study was to investigate changes in EEG power spectrum due to localized exposure in different parts of the brain by extremely low-frequency magnetic fields (ELF-MFs) to extract some protocols for treatment of some psychological disorders. In addition, regular effects were investigated by increasing intensity of ELF-MF. Therefore, EEG relative power spectrum was evaluated at T4, T3, F3, F4, and Cz points, when all the points were exposed to MFs with 45, 17, 10, 5, and 3 Hz frequencies, separately. Intensity of MF was 0, 100, 240, or 360 µT in four sessions. Significant changes were observed in different EEG bands caused by locally exposing to ELF-MF in different points of brain (P < 0.05). Some exposure to MFs decreased alpha band of frontal and central areas in closed-eyes state. Based on the findings in this study, some protocols can be designed using a combination of various MFs exposures to conduct the brain signals that is necessary to evaluate clinically.

Local exposure of brain central areas to a pulsed ELF magnetic field for a purposeful change in EEG.

This study examines the simultaneous exposure of 2 brain areas in the location of central electrodes (C3 and C4) to a weak and pulsed extremely low-frequency magnetic field (ELF-MF) on the electroencephalogram (EEG). The intent is to change the EEG for a therapeutic application, such as neurofeedback, by inducing the "resonance effect." A total of 10 healthy women received 9 minutes of ELF-MF (intensity 200 µT) and sham in a counterbalanced design. ELF-MF exposure frequencies were 10, 14, and 18 Hz. The paired t test revealed that local pulsed ELF-MF significantly decreases beta (15-25 Hz), sensorimotor rhythm (13-15 Hz), and theta (4-8 Hz) powers at a frequency of 10 Hz in C3 and C4 regions (12.0%-26.6%) after exposure, in comparison with that achieved during the exposure (P < .05). Variations during the exposure were transient and different from those after. The resonance effect was observed nowhere around the regions. The study suggests that this technique may be applied in the treatment of anxiety; however, further investigation is needed.

Study of the frequency parameters of EEG influenced by zone-dependent local ELF-MF exposure on the human head.

It has been reported that human subjects exposed to electromagnetic fields exhibit changes in human EEG signals at the frequency of stimulation. The aim of the present study was to expose different parts of the brain to extremely low-frequency magnetic fields locally and investigate EEG power spectrum alters at the frequency of stimulation. EEG relative power spectrum were evaluated at 3, 5, 10, 17, and 45 Hz frequencies at T4, T3, F3, Cz, and F4 points, respectively, when these points were exposed to magnetic fields with similar frequencies and 100 µT intensity. The paired t-test results showed that power value of EEG did not alter significantly at the frequency of stimulation (P<0.05). Further, significant changes in different EEG bands caused by locally exposing to ELF-MF in different points of brain were observed. The changes in the EEG bands were not limited necessarily to the exposure point.