Correlative fractography: Combining scanning electron microscopy and light microscopes for qualitative and quantitative analysis of fracture surfaces

Correlative fractography is a new expression proposed here to describe a new method for the association between scanning electron microscopy (SEM) and light microscopy (LM) for the qualitative and quantitative analysis of fracture surfaces. This article presents a new method involving the fusion of one elevation map obtained by extended depth from focus reconstruction from LM with exactly the same area by SEM and associated techniques, as X-ray mapping. The true topographic information is perfectly associated to local fracture mechanisms with this new technique, presented here as an alternative to stereo-pair reconstruction for the investigation of fractured components. The great advantage of this technique resides in the possibility of combining any imaging methods associated with LM and SEM for the same observed field from fracture surface. © Microscopy Society of America 2013.

Photodynamic inactivation of Streptococcus mutans and Streptococcus sanguinis biofilms in vitro

The purpose of this study was to evaluate specific effects of photodynamic inactivation (PDI) using erythrosine (ER) and Rose Bengal (RB) photosensitizers and a blue light-emitting diode (LED) on the viability of Streptococcus mutans and Streptococcus sanguinis biofilms. Biofilms were grown in acrylic disks immersed in broth to production of biofilms, inoculated with microbial suspension (106 cells/mL) and incubated for 48 h. After the formation of biofilms, the effects of the photosensitizers ER and RB at a concentration of 5 μM for 5 min and blue LED (455 ± 20 nm) for 180 s, photosensitizers alone and conjugated were evaluated. Next, the disks were placed in tubes with sterile physiological solution (0.9 % sodium chloride) and sonicated for to disperse the biofilms. Tenfold serial dilutions were carried and aliquots seeded in brain heart infusion agar which were then incubated for 48 h. Then the numbers colony-forming units per milliliter (CFU/mL; log 10) were counted and analyzed statistically (ANOVA, Tukey test, P ≤ 0.05). Significant decreases in the viability of all microorganisms were observed for biofilms exposed to PDI mediated by both photosensitizers. The reductions with RB and ER were, 0.62 and 0.52 log10 CFU mL -1 for S. mutans biofilms (p = 0.001), and 0.95 and 0.88 log 10 CFU mL-1 for S. sanguinis biofilms (p = 0.001), respectively. The results showed that biofilms formed in vitro by S. mutans and S. sanguinis, were sensitive to PDI using a blue LED associated with photosensitizers ER or RB, indicating its use in the control of caries and periodontal diseases. © 2012 Springer-Verlag London Ltd.

Foliar and Radicular Absorption of Boron by Beetroot and Tomato Plants

The isotopic technique was employed to study boron (B) mobility in tomato and beetroot plants under protected cultivation conditions. An experiment was conducted in which both species grew in 10-dm3 vases filled with coconut fiber, under hydroponic conditions. The plants were subjected to four different treatments: (1) no B in the substratum and no foliar fertilization; (2) no B in the substratum, with foliar 10B fertilization; (3) B in the substratum, with foliar 10B fertilization; and (4) 10B in the substratum and no foliar fertilization. The biological growth variables and total B and 10B contents in the plant parts grown after the application of the nutrient were evaluated. For increasing B content in young tissues, the foliar application of this element was not as efficient as application via root system, indicating low mobility of B in the tissues of both beetroot and tomato plants. © 2013 Copyright Taylor & Francis Group, LLC.

Characterization and impact of the sugarcane borer on sugarcane yield and quality

The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), is a key pest of sugarcane (Saccharum spp.). While damage caused by this pest has increased in the past 20 yr, studies investigating the insect-plant interactions are still lacking. Moreover, there is no information about the consequences of borer damage on the parameters of sugar quality. Therefore, two field experiments were performed during the 2010 and 2011 growing seasons in Brazil to compare the raw material and sugar quality of SP80-3280 sugarcane plants with and without the sugarcane borer. Plants were protected within screen cages and infested weekly during the 2010 and 2011 seasons, using egg masses starting at the second and third internode stage. At harvest, 25.77 and 19.01% of the internodes were bored by larvae (infestation intensity, II) in the first and second seasons, respectively. There was no correlation between the borer gallery total volume and II. The fiber content significantly increased with increasing II. The stalk biometric parameters, such as length, diameter, and yield, were not correlated with II. The sucrose yield significantly decreased with increasing II. Consequently, sugar yield losses were estimated at 8.83 and 19.80% per 1% bored internode for the first and second seasons, respectively. The concentration of phenolic compounds increased, and unclarified juice color quality decreased, with increasing II. Significant differences were detected in the quality of the sugar. These results should be confirmed for other sugarcane cultivars and incorporated into an economic injury level to enhance decision-making strategies for borer management. © 2013 by the American Society of Agronomy, 5585 Guilford Road, Madison, WI 53711. All rights reserved.

Effect of intercropping on yields of corn with different relative maturities and Palisadegrass

Intercropping corn (Zea mays L.) with palisadegrass [Brachiaria brizantha (Hochst. ex A. Rich) Stapf] can result in high amounts of residue and improve nutrient cycling. Long-season corn hybrids will live longer, competing with palisadegrass, which may reduce both corn and forage biomass yields. This study, conducted in the state of São Paulo, Brazil, had the objective of evaluating nutrient concentration and yield of corn hybrids with different maturity ratings as affected by intercropped palisadegrass as well as forage dry matter production. Te experimental design was randomized blocks with a factorial arrangement of eight treatments consisting of two cropping systems (corn alone and intercropped with palisadegrass) and four corn hybrids (105-, 121-, 132, and 144-d relative maturity). Compared with corn grown alone, intercropping treatments resulted in corn grain yields of 107% (105-d hybrid) to 71.7% (144-d hybrid). In the corn-alone system, the 132- and 144-d corn hybrids provided the highest corn yields (9581 and 9606 kg ha-1, respectively). Corn yield was similar between the single-crop and intercrop systems when using 105-, 121-, and 132-d hybrids. Intercropping with the 144-d hybrid reduced forage production (6619 kg ha-1) and quality of palisadegrass (86 g kg-1 of crude protein) compared with the other hybrids. Te intercropping system with the 132-d hybrid allowed both the highest corn grain (8860 kg ha-1) and palisadegrass (8256 kg ha-1) yields. Therefore, intercropping palisadegrass with the earlier (105-, 121-, and 132-d) corn hybrids is a viable option for crop-livestock integration because it did not affect either corn or palisadegrass yield. © 2013 by the American Society of Agronomy, 5585 Guilford Road, Madison, WI 53711. All rights reserved.

Association between IL8 haplotypes and pathogen levels in chronic periodontitis

Chronic periodontitis (CP) is considered to be a multifactorial disease influenced by microbial and genetic factors. The aim of the present study was to investigate whether the genetic susceptibility to CP in individuals with the IL8 ATC/TTC haplotype is associated with subgingival levels of periodontopathogens. Sixty-five individuals, grouped according to the presence (n = 28) or absence (n = 37) of the IL8 haplotype, were evaluated. After clinical periodontal evaluation, each group was subdivided according to the presence (CP) or absence (H) of periodontitis. Four subgingival samples were obtained from CP and two samples per subject from H patients. The levels and proportions of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were analyzed using quantitative real-time polymerase chain reaction (q-PCR). No differences were found in the proportion of periodontopathogenic bacteria between groups with the presence or absence of the IL8 haplotype. However, in the CP groups, the levels of periodontopathogens were significantly higher in the individuals without the IL8 haplotype than in the individuals with the IL8 haplotype. These results suggest that periodontal destruction may occur in patients who are considered to be genetically susceptible to CP with a lower microbial challenge because of the presence of the IL8 ATC/TTC haplotype than in patients without this haplotype. © 2013 Springer-Verlag Berlin Heidelberg.

Photodynamic antimicrobial therapy of curcumin in biofilms and carious dentine

Photodynamic therapy (PDT) is a technique that involves the activation of photosensitizers by light in the presence of oxygen, resulting in the production of reactive radicals that are capable of inducing cell death. The present study evaluated the susceptibility of Streptococcus mutans and Lactobacillus acidophilus to PDT grown as multi-species in the biofilm phase versus in dentine carious lesions. A brain-heart infusion culture medium supplemented with 1 % glucose, 2 % sucrose, and 1 % young primary culture of L. acidophilus 108 CFU/mL and S. mutans 108 CFU/mL was used to develop multi-species biofilms and to induce caries on human dentine slabs. Five different concentrations of curcumin (0.75, 1.5, 3.0, 4.0, and 5.0 g/L) were used associated with 5.7 J/cm2 light emission diode. Four different groups were analyzed L-D- (control group), L-D+ (drug group), L+D- (light group), and L+D+ (PDT group). ANOVA/Tukey's tests were conducted to compare groups. A significant reduction (p <0.05) in cell viability was observed in the biofilm phase following photosensitization with all curcumin concentrations tested. To achieve significant bacterial reduction (p <0.05) in carious dentine, it was necessary to utilize 5.0 g/L of curcumin in association with blue light. No significant reduction was found for L-D+, supporting the absence of the drug's dark toxicity. S. mutans and L. acidophilus were susceptible to curcumin in the presence of blue light. However, due to light penetration and drug diffusion difficulties, these microorganisms within dentine carious lesions were less affected than they were in the biofilm phase. © 2013 Springer-Verlag London.

Effects of soil and foliar application of soluble silicon on mineral nutrition, gas exchange, and growth of potato plants

Silicon can alleviate biotic and abiotic stresses in several crops, and it has beneficial effects on plants under nonstressed conditions. However, there is still doubt about foliar-applied Si efficiency and Si effects on mineral nutrition, physiological processes, and growth of potato (Solanum tuberosum L.) plants under wellwatered conditions. The objective of this study was to evaluate the effect of soil and foliar application of soluble Si on Si accumulation, nutrients, and pigments concentration as well as gas exchange and growth of potato plants. The experiment was conducted under greenhouse conditions in pots containing 35 dm3 of a Typic Acrortox soil. The treatments consisted of a control (no Si application), soil application of soluble Si (50 mg dm-3 Si), and foliar application of soluble Si (three sprays of 1.425 mM Si water solution, prepared with a soluble concentrate stabilized silicic acid), with eight replications. Both soil and foliar application of Si resulted in higher Si accumulation in the whole plant. Foliar application of Si resulted in the greatest Si concentration in leaves, and soil application increased Si concentration in leaves, stems, and roots. Silicon application, regardless of the application method, increased leaf area, specific leaf area, and pigment concentration (chlorophyll a and carotenoids) as well as photosynthesis and transpiration rates of wellwatered potato plants. However, only soil application increased P concentration in leaves and dry weight of leaves and stems. © Crop Science Society of America.

Cryptococcosis: epidemiology, fungal resistance, and new alternatives for treatment

Cryptococcosis is an important systemic mycosis and the third most prevalent disease in human immunodeficiency virus (HIV)-positive individuals. The incidence of cryptococcosis is high among the 25 million people with HIV/acquired immunodeficiency syndrome (AIDS), with recent estimates indicating that there are one million cases of cryptococcal meningitis globally per year in AIDS patients. In Cryptococcus neoformans, resistance to azoles may be associated with alterations in the target enzyme encoded by the gene ERG11, lanosterol 14α-demethylase. These alterations are obtained through mutations, or by overexpressing the gene encoding. In addition, C. gattii and C. neoformans present a heteroresistance phenotype, which may be related to increased virulence. Other species beyond C. neoformans and C. gattii, such as C. laurentii, have been diagnosed mainly in patients with immunosuppression. Infections of C. albidus have been isolated in cats and marine mammals. Recent evidence suggests that the majority of infections produced by this pathogen are associated with biofilm growth, which is also related with increased resistance to antifungal agents. Therefore, there is a great need to search for alternative antifungal agents for these fungi. The search for new molecules is currently occurring from nanoparticle drugs of plant peptide origin. This article presents a brief review of the literature regarding the epidemiology of cryptococcosis, as well as fungal resistance and new alternatives for treatment. © 2013 Springer-Verlag Berlin Heidelberg.

Susceptibility of multispecies biofilm to photodynamic therapy using Photodithazine®

This in vitro study evaluated the effect of photodynamic therapy (PDT) on the multispecies biofilm of Candida albicans, Candida glabrata, and Streptococcus mutans. Standardized fungal and bacterial suspensions were cultivated appropriately for each species and inoculated in 96-well microtiter plates for mix-biofilm formation. After 48 h of incubation, the biofilms were submitted to PDT (P + L+) using Photodithazine® (PDZ) at 100, 150, 175, 200, or 250 mg/mL for 20 min and 37.5 J/cm2 of light-emitting diode (LED) (660 nm). Additional samples were treated only with PDZ (P + L-) or LED (P-L+), or neither (control, P-L-). Afterwards, the biofilms were evaluated by quantification of colonies (CFU/mL), metabolic activity (XTT reduction assay), total biomass (crystal violet staining), and confocal scanning laser microscopy (CSLM). Data were analyzed by one-way ANOVA and Tukey tests (p < 0.05). Compared with the control, PDT promoted a significant reduction in colonies viability of the three species evaluated with 175 and 200 mg/mL of PDZ. PDT also significantly reduced the metabolic activity of the biofilms compared with the control, despite the PDZ concentration. However, no significant difference was found in the total biomass of samples submitted or not to PDT. For all analysis, no significant difference was verified among P-L-, P + L-, and P-L+. CSLM showed a visual increase of dead cells after PDT. PDT-mediated PDZ was effective in reducing the cell viability of multispecies biofilm. © 2013 Springer-Verlag London.

Effect of antimicrobial photodynamic therapy on periodontally infected tooth sockets in rats

The aim of the present study was to evaluate the antimicrobial effect of antimicrobial photodynamic therapy (aPDT) in alveolar treatment of areas with induced periodontitis. Thirty male Wistar rats were subjected to ligature-induced periodontal disease (PD) in the first left inferior molars, while the right side molars did not receive ligatures. After 7 days of PD evolution, ligatures were removed from the left side, and the first left and right mandibular molars were extracted. Afterwards, animals were divided into groups according to the following treatments: control (C)-no treatment; mechanical debridement (MD)-mechanical debridement and irrigation with saline solution; and aPDT-mechanical debridement, irrigation with toluidine blue O (TBO), and 1 min of laser irradiation (GaAlAs, 660 nm, 30 mW, 32 J/cm2, 60 s). Ligatures were removed and samples of the alveolar content after extraction and after each treatment were collected for microbial processing by real-time polymerase chain reaction with specific primers for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Treponema denticola. Data were submitted to statistical analysis by multiple comparison tests (McNemar test; p < 0.05). T. denticola was not found in the collected samples. A. actinomycetemcomitans and P. gingivalis were found in ligature samples. Tooth socket samples without periodontitis induction presented lesser microbial charge than samples with induced periodontitis (p < 0.05). aPDT significantly reduced A. actinomycetemcomitans levels on the left side (p < 0.05). It was concluded that aPDT was an effective antimicrobial treatment for tooth sockets in areas affected by induced periodontitis. © 2013 Springer-Verlag London.

Mating in the pseudogamic apomictic Anemopaegma acutifolium DC: Another case of pseudo-self-compatibility in Bignoniaceae?

Self-compatibility in apomictic pseudogamic species is considered fundamental to assure reproduction by seeds in extreme situations, making apomictic species more advantageous than sexual ones in these scenarios. Anemopaegma acutifolium is a polyploidy, apomictic sporophytic species with no endosperm development in ovules of unpollinated pistils, which indicates obligate pseudogamy. Thus, the aim of the present work is to study the breeding system and post-pollination events to test if there is similar pseudogamous development irrespective of pollination treatment. We analysed fruit and seed set obtained in controlled experimental pollinations, as well as embryo number per seed, and the progress of ovule penetration, fertilisation and early endosperm development between self- and cross-pollinated pistils. We found that the species is self-fertile and that spontaneous selfing fruit set is also possible, although emasculated flowers never form fruits. Selfed pistils were as efficient as crossed ones for all parameters analysed, except for a delay in endosperm development observed in the former that may be an effect of the late-acting self-incompatibility. Therefore, the avoidance of selfed pistil abortion seems to be promoted by the presence of adventitious embryos and a normal endosperm. We conclude that A. acutifolium shows apomixis-related pseudo-self-compatibility, as in other self-fertile apomictic species of Bignoniaceae, which confer reproductive assurance and increases fruit-set and persistence ability in fast-changing tropical habitats. © 2012 German Botanical Society and The Royal Botanical Society of the Netherlands.

Soil organic matter and physical attributes affected by crop rotation under no-till

Growing cover crops in systems under no tillage affects different pools of soil organic matter, and eventually soil physical attributes are modified. The objective of this study was to evaluate changes in soil organic matter and their relationship with soil physical attributes as affected by plant species grown in rotation with soybean [Glycine max (L.) Merr.] under no-till for 3 yr. Crop rotations included grain sorghum [Sorghum bicolor (L.) Moench], ruzigrass [Urochloa ruziziensis (R. Germ, and CM. Evard) Crins] and sorghum mixed with ruzigrass, all grown in fall/winter, followed by pearl millet [Pennisetum americanum (L.) Leeke], sunn hemp (Crotalaria juncea L.) and sorghum-sudangrass [S. bicolor × S. sudanense (Piper) Stapf] grown during the spring, plus a fallow check plot. Soybean was grown as the summer crop. Millet and sorghum-sudangrass cropped in spring showed higher root and shoot production as spring cropping. In fall/winter, sorghum mixed with ruzigrass yielded higher phytomass compared with sole cropping. Soil physical attributes and organic matter fractioning were positively affected by cropping millet and sorghum-sudangrass whereas intermediate effects were observed after sunn hemp. Maintaining fallow in spring had negative effects on soil organic matter and physical properties. Ruzigrass and sorghum mixed with ruzigrass cropped in fall/winter resulted in better soil quality. Spring cover crops were more efficient in changing soil bulk density, porosity, and aggregates down to 0 to 10 cm; on the other hand, fall/winter cropping showed significant effects on bulk density in the uppermost soil layer. Total C levels in soil were increased after a 3-yr rotation period due to poor initial physical conditions. Fractions of particulate organic C, microbial C, and C in macroaggregates were the most affected by crop rotations, and showed high relation with improved soil physical attributes (porosity, density, and aggregates larger than 2 mm). © Soil Science Society of America, All rights reserved.

Comparison of the effect of rose bengal- and eosin Y-mediated photodynamic inactivation on planktonic cells and biofilms of Candida albicans

Candida albicans is an opportunistic yeast that can cause oral candidosis through the formation of a biofilm, an important virulence factor that compromises the action of antifungal agents. The objective of this study was to compare the effect of rose bengal (RB)- and eosin Y (EY)-mediated photodynamic inactivation (PDI) using a green light-emitting diode (LED; 532 ± 10 nm) on planktonic cells and biofilms of C. albicans (ATCC 18804). Planktonic cultures were treated with photosensitizers at concentrations ranging from 0.78 to 400 μM, and biofilms were treated with 200 μM of photosensitizers. The number of colony-forming unit per milliliter (CFU/mL) was compared by analysis of variance and Tukey's test (P ≤ 0.05). After treatment, one biofilm specimen of the control and PDI groups were examined by scanning electron microscopy. The photosensitizers (6.25, 25, 50, 200, and 400 μM of EY, and 6.25 μM of RB or higher) significantly reduced the number of CFU/mL in the PDI groups when compared to the control group. With respect to biofilm formation, RB- and EY-mediated PDI promoted reductions of 0.22 log10 and 0.45 log10, respectively. Scanning electron microscopy showed that the two photosensitizers reduced fungal structures. In conclusion, EY- and RB-mediated PDI using LED irradiation significantly reduced C. albicans planktonic cells and biofilms. © 2013 Springer-Verlag London.

Cover crop termination timing on rice crop production in a no-till system

Measuring shikimic acid accumulation in response to glyphosate applications can be a rapid and accurate way to quantify and predict glyphosate-induced damage to sensitive plants. The objective of this paper was to evaluate the effect of cover crop termination timing by glyphosate application on rice (Oryza sativa L.) yield in a no-till system. A factorial experiment, arranged in a split-plot design, was conducted for 2 yr. Treatments consisted of cover crops (main plots) and timed herbicide applications (subplots) to these cover crops (30, 20, 10, and 0 d before rice planting). There was a decrease in rice yield from 2866 kg ha-1 to 2322 kg ha-1 when the herbicide was applied closer to the rice planting day. Glyphosate application on cover crops increased shikimate concentrations in rice seedlings cultivated under palisade grass (Brachiaria brizantha), signal grass (B. ruziziensis), guinea grass (Panicum maximum), and weedy fallow (spontaneous vegetation) but not under millet (Pennisetum glaucum), which behaved similarly to the control (clean fallow, no glyphosate application). Glyphosate applications in the timing intervals used were associated with stress in the rice plants, and this association increased if cover crops took longer to completely dry and if higher amounts of biomass were produced. Millet, as a cover crop, allowed the highest seedling dry matter for upland rice and the highest rice yield. Our results suggest that using millet as a cover crop, with glyphosate application far from upland rice planting day (10 d or more), was the best option for upland rice under a no-tillage system. © Crop Science Society of America.