Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Resumo A esporulação de Didymella bryoniae in vitro é de grande importância para estudos que requerem inóculo puro e em grandes quantidades. Assim, os objetivos deste estudo foram identificar a melhor condição para esporulação de D. bryoniae combinando diferentes espectros de luz (luz UV-A ou UV-B, luz branca e escuro contínuo) com distintos meios de cultura (PDA, V8, ML e PDAB) e, avaliar a sobrevivência do fungo armazenado a -20°C ao longo do tempo. As amostras de fungo só esporularam quando submetidas ao tratamento com luz UV-B, independentemente do meio de cultura. Maior aparecimento de esporos do tipo conídio foi observado no meio PDAB, e a menor produção ocorreu no meio ML. Estruturas reprodutivas, como peritécios e picnídeos, foram observadas em todos os meios de cultura. No entanto, houve uma variação considerável na quantidade de cada estrutura entre os diferentes meios de cultura. Os meios ML e V8 apresentaram maior número de peritécios e os meios PDA e PDAB apresentaram maior proporção de picnídeos em relação aos peritécios. A duração do armazenamento a -20°C não afetou o crescimento micelial ou a taxa de crescimento micelial. Em conclusão, a luz UV-B é essencial para a esporulação de D. bryoniae in vitro. Além disso, a composição do meio de cultura influencia o tipo de estrutura fúngica produzida, bem como o tamanho e a quantidade dos esporos. O congelamento a -20°C é uma técnica eficiente que pode ser usada para armazenar D. bryoniae por pelo menos cinco meses sem perda de viabilidade

Copaiba oil and Neem extract can be a potential alternative for the behavioral control of Sitophilus zeamais

Abstract Insects' ethology is an important factor when it is desired to carry out pest management. This knowledge makes it possible to manipulate behavioral activities, repel, or attract insects according to needs and interests. The maize weevil Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), one of the main stored grain pests, has been the target of studies of behavioral changes studies through natural substances due to its resistance to different insecticidal classes. Thus, this study aimed to evaluate the effect of sublethal concentrations of neem extract and copaiba oil on the locomotor behavior of S. zeamais. The behavioral characteristic considered were walking activity, the frequency of contact of insects with the treated grain mass, and the time spent for this behavior. The walking activity of the S. zeamais increased with exposure to Neem extract and Copaiba oil. In general, the Neem extract and Copaiba oil-induced more contact with grain mass than the control, suggesting an attractive effect on the insect, however more significant for the Neem oil. The insect's behavior was altered, presenting a specific path due to Copaiba oil and Neem extract stimuli. These results indicate that Copaiba oil and Neem extract can be a potential alternative for controlling S. zeamais on stored products since changes in this pests' behavior can reduce qualitative and quantitative grain damage. Thus, the development of products based on Copaiba oil and Neem extract may be helpful for storage pest management.

Resumo A etologia dos insetos é um fator importante quando se deseja relizar manejo de pragas. Através deste conhecimento, é possível manipular atividades comportamentais, repelir, ou atrair os insetos de acordo com as necessidades e interesses. O gorgulho do milho Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), uma das principais pragas de grãos armazenados tem sido alvo de estudos de alterações comportamentais através de substâncias naturais devido à sua resistência a diferentes classes de insecticidas. Assim, este estudo teve como objetivo avaliar o efeito de concentrações subletais de extrato de neem e do óleo de copaíba sobre o comportamento de movimentação de S. zeamais. As características comportamentais consideradas foram: a atividade de caminhamento, a frequência do contato dos insetos com a massa de grãos tratada, e o tempo gasto para realização destes comportamentos. A atividade de caminhamento do S. zeamais aumentou quando os insetos foram expostos ao extrato de Neem e ao óleo de copaíba. Em geral, o extrato de Neem e o óleo de Copaíba induziram mais contato com a massa de grãos do que o controle, sugerindo um efeito atrativo sobre o inseto, contudo este efeito foi mais significativo para o óleo de Neem. O comportamento do inseto foi alterado, apresentando um caminhamento específico devido aos estímulos do óleo de copaíba e do extrato de Neem. Estes resultados indicam que o óleo de copaíba e o extrato de Neem podem ser alternativas potenciais para o controle do S. zeamais em produtos armazenados, uma vez que alterações no comportamento desta praga podem reduzir os danos qualitativos e quantitativos nos grãos. Assim, o desenvolvimento de produtos baseados no óleo de copaíba e no extrato de Neem pode ser útil para o manejo de pragas de armazenamento.

Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature / Fornecendo inóculo para estudos de Didymella bryoniae: o efeito do espectro de luz e armazenamento em baixa temperatura

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus' survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores' size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Resumo A esporulação de Didymella bryoniae in vitro é de grande importância para estudos que requerem inóculo puro e em grandes quantidades. Assim, os objetivos deste estudo foram identificar a melhor condição para esporulação de D. bryoniae combinando diferentes espectros de luz (luz UV-A ou UV-B, luz branca e escuro contínuo) com distintos meios de cultura (PDA, V8, ML e PDAB) e, avaliar a sobrevivência do fungo armazenado a -20°C ao longo do tempo. As amostras de fungo só esporularam quando submetidas ao tratamento com luz UV-B, independentemente do meio de cultura. Maior aparecimento de esporos do tipo conídio foi observado no meio PDAB, e a menor produção ocorreu no meio ML. Estruturas reprodutivas, como peritécios e picnídeos, foram observadas em todos os meios de cultura. No entanto, houve uma variação considerável na quantidade de cada estrutura entre os diferentes meios de cultura. Os meios ML e V8 apresentaram maior número de peritécios e os meios PDA e PDAB apresentaram maior proporção de picnídeos em relação aos peritécios. A duração do armazenamento a -20°C não afetou o crescimento micelial ou a taxa de crescimento micelial. Em conclusão, a luz UV-B é essencial para a esporulação de D. bryoniae in vitro. Além disso, a composição do meio de cultura influencia o tipo de estrutura fúngica produzida, bem como o tamanho e a quantidade dos esporos. O congelamento a -20°C é uma técnica eficiente que pode ser usada para armazenar D. bryoniae por pelo menos cinco meses sem perda de viabilidade

The Impact of Different Surface Treatments on Topography and Bond Strength of Resin Cement to Lithium Disilicate Glass Ceramic.

OBJECTIVE: To evaluate the influence of different surface treatments on topography, surface roughness, surface energy, and microtensile bond strength stability of resin cement to lithium disilicate glass ceramic. METHODS AND MATERIALS: Seventy disc-shaped specimens of IPS e.max Press were divided into seven groups according to the surface treatment: NT - non-treated (control); FSil - hydrofluoric acid (HF) + silane; FPSil - HF + phosphoric acid (HP) + silane; FUSil - HF + ultrasonic bath (U) + silane; FPUSil - HF + HP + U + silane; MEP - Monobond Etch and Prime (MEP); and MEPH - MEP + heating. Topography and surface roughness were evaluated using a 3D laser confocal microscope (3DLCM) and scanning electron microscopy (SEM) and surface energy with a goniometer. The microtensile bond strength (µTBS) was evaluated after storage in distilled water at 37°C for 24 hours and after thermocycling (5,000 cycles, 5°-55° C, 30-second dwell time). Data were analyzed using one-way ANOVA (surface roughness and surface energy), two-way ANOVA (µTBS), Tukey's HSD post-hoc test, and Student t-test (α=0.05). RESULTS: FUSil, FPSil, and FSil presented similar and highest surface roughness, whereas NT, FPUSil, MEP, and MEPH showed similar, and lowest, roughness values (p<0.05). FPUSil, FPSil, FUSil, and FSil presented a similar and highest surface energy. NT, MEP, and MEPH showed similar and lowest surface energy. CONCLUSION: Only FSil, FPSil, and MEPH maintained bond strength stability after thermocycling, with FPSil leading to less bond strength reduction, suggesting this protocol is more reliable for bonding resin cements to lithium disilicate glass ceramics.

Copaiba oil and Neem extract can be a potential alternative for the behavioral control of Sitophilus zeamais.

Insects' ethology is an important factor when it is desired to carry out pest management. This knowledge makes it possible to manipulate behavioral activities, repel, or attract insects according to needs and interests. The maize weevil Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), one of the main stored grain pests, has been the target of studies of behavioral changes studies through natural substances due to its resistance to different insecticidal classes. Thus, this study aimed to evaluate the effect of sublethal concentrations of neem extract and copaiba oil on the locomotor behavior of S. zeamais. The behavioral characteristic considered were walking activity, the frequency of contact of insects with the treated grain mass, and the time spent for this behavior. The walking activity of the S. zeamais increased with exposure to Neem extract and Copaiba oil. In general, the Neem extract and Copaiba oil-induced more contact with grain mass than the control, suggesting an attractive effect on the insect, however more significant for the Neem oil. The insect's behavior was altered, presenting a specific path due to Copaiba oil and Neem extract stimuli. These results indicate that Copaiba oil and Neem extract can be a potential alternative for controlling S. zeamais on stored products since changes in this pests' behavior can reduce qualitative and quantitative grain damage. Thus, the development of products based on Copaiba oil and Neem extract may be helpful for storage pest management.

Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature.

The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus' survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores' size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

COMT rs4818, pain sensitivity and duration, and alveolar bone grafting of oral clefts.

PURPOSE: Verifying whether the mutation in COMT rs4818 could be involved in pain modulation. METHODS: Thirty-two individuals born with cleft lip and palate that underwent bone graft from the iliac crest bone were assessed at 12, 24, 48, 72 h, and 7 days regarding their pain experience using a visual analogic scale. DNA from each participant was collected from saliva samples, and genotyping of rs4818 was performed using TaqMan chemistry. Overrepresentation of rs4818 alleles was tested using chi-square or Fisher's exact tests with an alpha of 0.05. RESULTS: Of the 32 individuals, eighteen reported long pain duration, nine reported high pain intensity, and fourteen low pain intensity up to 48 h. No differences were found in the distribution of individuals depending on the reported pain by sex (p = 0.12), age (p = 0.42), or cleft type (p = 0.5). The distribution of COMT r4818 alleles was different depending on the intensity and duration of pain. Carriers of the C wild-type allele were four times more likely to show high pain intensity and duration (odds ratio = 4.29, 95% confidence interval 1.13-16.18), meaning that the G variant allele is protective. CONCLUSION: COMT rs4818 is associated with postoperative pain after alveolar bone grafting.

Ultrastructural Aspects of The Intestinal Cells Surface of Angiostrongylus Cantonensis By Using of Transmission Electron Microscopy.

The rat lungworm Angiostrongylus cantonensis is a zoonotic parasite and the main cause of eosinophilic meningitis in the world. Its main source of nutrients originates from the degradation of host hemoglobin in blood-feeding helminths, such as A. cantonensis. The purpose of this study was to analyze the ultrastructure of the intestinal cells by using of transmission electron microscopy (TEM). A. cantonensis worms obtained from Rattus norvegicus (norway rats) from endemic area were used for TEM. The ultrastructural analysis was performed using fragments cut from the middle part of the worms, and the TEM study revealed the cells with microvilli and nuclei containing areas of loose and condensed chromatin and the presence of macrovesicles and microvesicles of digestion and it was observed in this study that intestinal epithelium is look like a syncytium. The ultrastructural aspects of the intestinal cells surface of A. Cantonensis the indicate that the intestinal epithelium is a multinucleate mosaic or syncytium.

Characterization of novel human intragenic antimicrobial peptides, incorporation and release studies from ureasil-polyether hybrid matrix.

Intragenic antimicrobial peptides (IAPs) are internal sequences of proteins with physicochemical similarities to Antimicrobial Peptides (AMPs) that, once identified and synthesized as individual entities, present antimicrobial activity. Many mature proteins encoded by the genomes of virtually any organism may be regarded as inner reservoirs of IAPs, conferring them ample biotechnological potential. However, IAPs may also share shortcomings with AMPs, such as low half-life in biological media and non-specific adsorption in eukaryotic cells. The present manuscript reports a translational approach that encompasses the uncovering of two novel IAPs from human proteins as well as the first results concerning the incorporation and sustained release of one of these peptides from ureasil-polyether hybrid polymeric films. For such, the software Kamal was used to scan putative IAPs in the human proteome, and two peptides, named Hs05 and Hs06, were identified, synthesized, and tested as antimicrobials. Biophysical assays were conducted using model phospholipid vesicles and 1H NMR solution structures in phospholipid micelles were obtained for the IAP Hs05. This peptide was incorporated in a polymeric matrix composed of the ureasil/PPO-PEO-PPO triblock copolymer, and the resulting films were evaluated by atomic force microscopy and imaging mass spectrometry. The release rate of Hs05 from the polymeric matrix was assessed and the antimicrobial activity of Hs05-loaded hybrid polymeric films was evaluated against the bacterium Escherichia coli. This study represents the first steps towards the development of polymeric films enriched with IAPs obtained from the human proteome as sustained release devices for topical application.

Crack cocaine smoke on pregnant rats: Maternal evaluation and teratogenic effect.

This study aimed to evaluate maternal toxicity, teratogenic, and placental oxidative effects resulting from the exposure of rats to crack cocaine smoke during pregnancy. Pregnant rats were exposed either to the smoke of crack and ashes (Crack) or to the smoke of ashes alone, nonexposed or pair-fed with the Crack group. Crack group was exposed to the smoke resulting from the burning of 250 mg of crack for 10 min, twice a day, from 7 days prior to mating until cesarian on gestational day 20. Placental oxidative stress and classical parameters of maternal and fetal evaluation were studied, in addition to the morphometric analysis of the fetal metamers. Even in the absence of changes in body weight gain and feed intake, crack altered the reproductive performance of dams. Exposure to the drug promoted late closure of the fetal fontanel. Furthermore, the morphometric study of the brain mass (BM)/skull cap ratio revealed a decrease in the BM of the fetuses exposed to the drug. Exposure to crack has an oxidative potential in fetal development, since exposure to the drug promoted placental lipid peroxidation. Our study showed that daily exposure to crack, even in lower frequency than that performed by users, has a teratogenic potential.

Poor survival after resection of early gastric cancer: extremes of survivorship analysis reveal distinct genomic profile.

A subset of patients with early gastric cancer demonstrate early recurrence and poor survival despite margin-negative resection. This study used an extremes-of-survivorship approach to identify an association between TP53 hotspot mutations co-occurring with loss of heterozygosity and unexpectedly poor survival in early gastric cancer. This distinct genomic profile may be a novel biomarker of poor survival in patients with completely resected early gastric cancer, and warrants large-scale validation. Promising, validation needed.

Effectiveness and safety of Omalizumab in the treatment of chronic spontaneous urticaria: Systematic review and meta-analysis.

INTRODUCTION: Chronic spontaneous urticaria (CSU) affects approximately 1% of the population, affecting both children and adults. Omalizumab (Oma) is a therapeutic option for patients with refractory forms of CSU. OBJECTIVES: To determine the effectiveness and safety of Oma in the treatment of CSU. METHODS: Systematic review (Cochrane Collaboration methodology) of randomized clinical trials comparing Oma to placebo in refractory CSU treatment. The search is based on MEDLINE; EMBASE, Central Cochrane Library, and LILACS. The outcomes evaluated were: control of the illness, adverse events, and quality of life. RESULTS: Of the 848 identified studies 13 were selected for further review and six were included in the meta-analysis. For all outcomes, high-quality evidence has confirmed that Oma is effective in the treatment of CSU. The dosage of 300mg/month achieved better results; namely a significant reduction in pruritus, papules, and urticaria activity, as well as an increase in the number of patients with a controlled condition, improvement in the quality of life and no differences in adverse events compared to the placebo. CONCLUSIONS: High-quality evidence demonstrates that Oma is effective and safe in the treatment of CSU refractory to therapy with H1 antihistamines.

Bond Strength Stability of Self-adhesive Resin Cement to Etched Vitrified Yttria-stabilized Tetragonal Zirconia Polycrystal Ceramic After Thermomechanical Cycling.

The purpose of this study was to evaluate the influence of thermomechanical cycling on the bond strength stability of self-adhesive resin cement to vitrified yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) ceramic. Y-TZP ceramic blocks were divided into four groups according to the surface treatments: AS, as sintered; SB, sandblasted with 50-µm Al2O3 particles; LG, vitrification with a low-fusing glaze and etching with 10% hydrofluoric acid; and HC, vitrification with a ceramic liner and etching with 10% hydrofluoric acid. Lithium disilicate ceramic blocks etched with 10% hydrofluoric acid for 20 seconds (EM) were also tested. Blocks of resin composite were cemented on the ceramic block surfaces using a self-adhesive resin cement. Half of the blocks from each group were cut into beams and submitted to microtensile bond strength (µTBS) testing after immersion in water at 37°C for 24 hours and the other half after mechanical (1,200,000 cycles, 98 N, 2.5 Hz) and thermal (10,000 cycles, 5-55°C, 30-second dwell time) cycling (TMC). Data were analyzed using one-way analysis of variance, Tukey's honest significant difference post hoc test, and Student t-test (α=0.05). After 24 hours, EM and LG presented similar and the highest µTBS, whereas AS showed the lowest µTBS (p<0.05). After TMC, EM presented the highest µTBS, followed by LG (p<0.05). Only EM and LG maintained the stability of µTBS after TMC (p>0.05). The bonding to Y-TZP ceramic vitrified with a low-fusing glaze withstood thermomechanical cycling.

Low-level laser therapy and anesthetic infiltration for orofacial pain in patients with fibromyalgia: a randomized clinical trial.

BACKGROUND: To compare the analgesic effect of anesthetic infiltration of lidocaine 2% and low-level laser therapy (LLLT) by GaAlAs into tender points of patients with orofacial pain and fibromyalgia (FM). MATERIAL AND METHODS: A randomized clinical trial was performed with adults (N=66) that were allocated into two groups (1:1): Group A received LLLT irradiation by Diode Laser GaAlAs (780nm) with expositions twice a week during six weeks and Group B was treated with anesthetic infiltration of lidocaine 2% without vasoconstrictor once a week for four weeks. The pain assessment included the Visual Analogic Scale (VAS) and tenderness to palpation. RESULTS: No dropout and adverse effect was observed during the study. The pain decreased significantly in each group after the treatment (p=0.0001, ß=1.0), even though no statistical difference was found between both treatments (p=0.46, ß= 0.82). The presence of tender points decreased after both treatments, with responsively in some types of masticatory muscles (p<0.05) except posterior temporalis muscle. The patients perception showed that both treatments were effective and a few patients reported that the treatment did not improve welfare. CONCLUSIONS: The LLLT by GaAlAs and anesthetic infiltration of lidocaine 2% were equally effective to control orofacial pain in FM individuals.

Effect of feeding strategies and cropping systems on greenhouse gas emission from Wisconsin certified organic dairy farms.

Organic agriculture continues to expand in the United States, both in total hectares and market share. However, management practices used by dairy organic producers, and their resulting environmental impacts, vary across farms. This study used a partial life cycle assessment approach to estimate the effect of different feeding strategies and associated crop production on greenhouse gas emissions (GHG) from Wisconsin certified organic dairy farms. Field and livestock-driven emissions were calculated using 2 data sets. One was a 20-yr data set from the Wisconsin Integrated Cropping System Trial documenting management inputs, crop and pasture yields, and soil characteristics, used to estimate field-level emissions from land associated with feed production (row crop and pasture), including N2O and soil carbon sequestration. The other was a data set summarizing organic farm management in Wisconsin, which was used to estimate replacement heifer emission (CO2 equivalents), enteric methane (CH4), and manure management (N2O and CH4). Three combinations of corn grain (CG) and soybean (SB) as concentrate (all corn = 100% CG; baseline = 75% CG + 25% SB; half corn = 50% CG + 50% SB) were assigned to each of 4 representative management strategies as determined by survey data. Overall, GHG emissions associated with crop production was 1,297 ± 136 kg of CO2 equivalents/t of ECM without accounting for soil carbon changes (ΔSC), and GHG emission with ΔSC was 1,457 ± 111 kg of CO2 equivalents/t of ECM, with greater reliance on pasture resulting in less ΔSC. Higher levels of milk production were a major driver associated with reduction in GHG emission per metric tonne of ECM. Emissions per metric tonne of ECM increased with increasing proportion of SB in the ration; however, including SB in the crop rotation decreased N2O emission per metric tonne of ECM from cropland due to lower applications of organically approved N fertility inputs. More SB at the expense of CG in the ration reduced enteric CH4 emission per metric tonne of ECM (because of greater dietary fat content) but increased N2O emission per metric tonne of ECM from manure (because of greater N content). An increased reliance on pasture for feed at the expense of grain resulted in decreased in milk production, subsequently leading to substantially higher emissions per metric tonne of ECM.

Características produtivas e nutricionais do capim-xaraés inoculado com bactérias diazotróficas associativas / Productive and nutritional characteristics of Xaraés grass inoculated with diazotrophic associative bacteria

O objetivo do trabalho foi avaliar as características produtivas e nutricionais do capim- xaraés inoculado com bactérias diazotróficas associativas, em primeiro cultivo em Latossolo Vermelho do Cerrado. O experimento foi realizado em casa de vegetação. O delineamento experimental foi inteiramente ao acaso, constituído por fatorial 6x3: três estirpes de bactérias diazotróficas associativas (MTAz8, MTH2 e Y2), uma combinação das estirpes MTAz8 e MTH2, adubação nitrogenada e testemunha absoluta, e três cortes (30, 60 e 90 dias), em cinco repetições. A inoculação foi feita por meio da inserção de uma alíquota de 5mL de caldo bacteriano contendo 108 células mL-1 no solo próximo ao sistema radicular. As variáveis avaliadas foram: massa seca da parte aérea, porcentagem de proteína bruta, acúmulo e concentração de nitrogênio na parte aérea e determinação do número de bactérias diazotróficas do solo. No tratamento adubado com nitrogênio, a produção de massa seca aumentou ao longo dos cortes. Entre as estirpes de bactérias associativas, a Y2 apresentou maior acúmulo de nitrogênio e maior população de bactérias no meio de cultura LGI. As características nutricionais do capim-xaraés decaíram à medida que foram realizados os cortes.(AU)

The aim of this study was to evaluate yield and nutritional characteristics of Xaraés grass inoculated with diazotrophic associative bacteria in the first cultivation in the Cerrado Oxisol. The experiment was conducted in the greenhouse. The experimental design was completely randomized factorial 6x3 consisting of: three strains of diazotrophic associative bacteria (MTAz8, MTH2 and Y2), a combination of strains MTAz8 and MTH2, nitrogen fertilization and absolute control, and three cuts (30, 60 and 90 days) in five replicates. The inoculation was performed by inserting an aliquot of 5ml of the bacterial broth containing 10 8 cells ml-1 in the soil near the root zone of each plant. The variables evaluated were: dry weight of shoot, crude protein, accumulation and nitrogen concentration in the shoot and determining the number of nitrogen-fixing bacteria in the soil. The fertilized soil dry matter production increased over the cuts. Among the strains Y2 showed higher accumulation of nitrogen and largest population of bacteria in the middle of the LGI culture. The nutritional characteristics of Xaraés grass declined as the cuts were made.(AU)

Remineralizing potential of CPP-ACP creams with and without fluoride in artificial enamel lesions.

BACKGROUND: This study evaluated the effect of pastes containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) with and without fluoride on enamel demineralization. METHODS: Human enamel blocks were used and after incipient caries-like lesions were formed, they were assigned to four groups: G1 - saliva; G2 - MI Paste (RecaldentTM ); G3 - MI Paste Plus (RecaldentTM 900 ppm as NaF); and G4 - Crest™ (1.100 ppm as NaF). The specimens were soaked in demineralizing solution for 6 hours and remineralized in artificial saliva for 18 hours alternatively for 10 days. The dentifrice was prepared with deionized water in a 1:3 proportion (w/w) or applied undiluted in the case of the CPP-ACP formula. Demineralized enamel changes were analysed by surface microhardness (SMH), 3D-profilometry and SEM. Data were analysed by non-parametric Kruskal-Wallis and Mann-Whitney test comparisons and one-way ANOVA and Tukey's HSD post hoc test (α = 0.05). RESULTS: The SMH values observed in the G2 (47.8 ± 28.5) and G3 (53.6 ± 27.6) groups were different from that of G4 (90.2 ± 17.1), which were significantly higher than that found in G1 (39.4 ± 14.2). The %SMH was significantly lower in G4 when compared to G1 (p < 0.001) and G3 (p < 0.05). CONCLUSIONS: MI Paste Plus demonstrated a greater protective effect against demineralization than MI Paste and showed smoother surfaces.

Avaliação das atividades antibacteriana, tripanocida e citotóxica do extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda (Veludo Branco) (Commelinaceae) / Evaluation of antibacterial, trypanocidal and cytotoxical activities of the hydroalcoholic extract from the roots of Tradescantia sillamontana Matuda (Veludo Branco) (Commelinaceae)

RESUMO O objetivo deste estudo foi avaliar o efeito antibacteriano e tripanocida in vitro do extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda (Commelinaceae), conhecida popularmente como veludo branco. Foi avaliada a atividade antibacteriana in vitro frente às bactérias Streptococcus mitis (CIM = 100 µg/mL; CMB = 150 µg/mL), Streptococcus mutans (CIM = 200 µg/mL; CMB = 220 µg/mL), Streptococcus sanguinis (CIM = 400 µg/mL; CMB = 425 µg/mL), Streptococcus sobrinus (CIM = 400 µg/mL; CMB = 420 µg/mL) e Bacteroides fragilis (CIM = 400 µg/mL; CMB = 430 µg/mL) pelo método de diluição em caldo. Os protozoários da família tripanossomatídeo causam doenças tropicais que costumam ser negligenciadas que costumam ser como a tripanossomíase, para a qual estão disponíveis poucos medicamentos. Neste contexto, o extrato hidroalcóolico das raízes de T. sillamontana também foi avaliado frente às formas tripomastigotas da cepa Y de Trypanosoma cruzi, com promissora atividade frente a este protozoário (IC50 = 2,4 µg/mL). Quando avaliada a atividade citotóxica frente a fibroblastos da linhagem LLCMK2, o extrato apresentou moderada citotoxicidade (CC50 = 480,37 µg/mL). Os resultados ora apresentados para o extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda demonstraram promissoras atividades antibacteriana e tripanocida, sendo uma fonte alternativa de produtos naturais com atividades contra T. cruzi e algumas bactérias do gênero Streptococcus e Bacteroides.

ABSTRACT The aim of this study was to investigate the in vitro, antibacterial and trypanocidal effect of the hydroalcoholic extract from the roots of Tradescantia sillamontana Matuda (Commelinaceae), commonly known as Veludo branco. The in vitro antibacterial activity against the standard bacteria Streptococcus mitis (MIC = 100µg/mL; MBC = 150 µg/mL), Streptococcus mutans (MIC = 200µg/mL; MBC = 220 µg/mL), Streptococcus sanguinis (MIC = 400µg/mL; MBC = 425 µg/mL), Streptococcus sobrinus (MIC = 400µg/mL; MBC = 420 µg/mL) andBacteroides fragilis (MIC = 400µg/mL; MBC = 430 µg/mL), using microdilution broth methods. Protozoans from the trypanosomatid family cause neglected tropical diseases such as trypanosomiasis, for which few drugs are available. In this context, the hydroalcoholic extract of the Tradescantia sillamontana roots was also investigated with regards to the in vitro effects against the trypomastigote forms of theY strain of Trypanosoma cruzi, showing strong activity against this parasite (IC50 = 2.4 µg/mL). When performing cytotoxic activity against fibroblasts LLCMK2 line, the extract showed moderate cytotoxicity (CC50 = 480.37 mg/mL). The results presented for the hydroalcoholic extract of the roots of Tradescantia sillamontana Matuda demonstrated effective antibacterial and trypanocidal activities and were shown to be an alternative source of natural products with activity against T. cruzi and some bacteria of the genus Streptococcus and Bacteroides.

Hemocytes of Rhipicephalus sanguineus (Acari: Ixodidae): Characterization, Population Abundance, and Ultrastructural Changes Following Challenge with Leishmania infantum.

Few studies have examined the cellular immune response of ticks, and further research on the characterization of the hemocytes of ticks is required, particularly on those of Rhipicephalus sanguineus (Latreille) because of the medical and veterinary importance of this tick. The aims of this study were to characterize the morphology and the ultrastructure of the different types of hemocytes of adult R. sanguineus and to determine the population abundance and the ultrastructural changes in the hemocytes of ticks infected with Leishmania infantum. The hemocytes were characterized through light and transmission electron microscopy. Within the variability of circulating cells in the hemolymph of adult R. sanguineus, five cell types were identified, which were the prohemocytes, plasmatocytes, granulocytes, spherulocytes, and adipohemocytes. The prohemocytes were the smallest cells found in the hemolymph. The plasmatocytes had polymorphic morphology with vesicles and cytoplasmic projections. The granulocytes had an elliptical shape with the cytoplasm filled with granules of different sizes and electrodensities. The spherulocytes were characterized by several spherules of uniform shapes and sizes that filled the entire cytoplasm, whereas the adipohemocytes had an irregular shape with multiple lipid inclusions that occupied almost the entire cytoplasmic space. The total counts of the hemocyte population increased in the group that was infected with L. infantum. Among the different cell types, the numbers increased and the ultrastructural changes occurred in the granulocytes and the plasmatocytes in the infected group of ticks.