Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Transient expression of a green fluorescent protein in tobacco and maize chloroplast

BACKGROUND: Maize is one of the most important crops worldwide and has been a target of nuclear-based transformation biotechnology to improve it and satisfy the food demand of the ever-growing global population. However, the maize plastid transformation has not been accomplished due to the recalcitrant condition of the crop. RESULTS: In this study, we constructed two different vectors with homologous recombination sequences from maize (Zea mays var. LPC13) and grass (Bouteloua gracilis var. ex Steud) (pZmcpGFP and pBgcpGFP, respectively). Both vectors were designed to integrate into rrn23S/rrn16S from an inverted repeat region in the chloroplast genome. Moreover, the vector had the mgfp5 gene driven by Prrn, a leader sequence of the atpB gene and a terminator sequence from the rbcL gene. Also, constructs have an hph gene as a selection marker gene driven by Prrn, a leader sequence from rbcL gene and a terminator sequence from the rbcL gene. Explants of maize, tobacco and Escherichia coli cells were transformed with both vectors to evaluate the transitory expression­an exhibition of green and red fluorescent light under epifluorescence microscopy. These results showed that both vectors were expressed; the reporter gene in all three organisms confirmed the capacity of the vectors to express genes in the cell compartments. CONCLUSIONS: This paper is the first report of transient expression of GFP in maize embryos and offers new information for genetically improving recalcitrant crops; it also opens new possibilities for the improvement in maize chloroplast transformation with these vectors.

Intrastromal voriconazole: An adjuvant approach for recalcitrant mycotic keratitis

Purpose: To evaluate the efficacy of intrastromal voriconazole for the management of fungal keratitis not responding to conventional therapy. Methods: Patients having microbiologically proven fungal keratitis with poor response to 2 weeks of conventional topical therapy were included in the study. After obtaining informed consent, an intrastromal injection of voriconazole was administered around the ulcer. Response to treatment in the form reduction in the size of the ulcer and infiltration was recorded on regular follow-ups. Results: Out of a total of 20 patients, 14 responded to intrastromal treatment and resolved, whereas six patients progressed to perforation. Mean resolution time was 35.5 � 9.2 days. The most common organism isolated was Fusarium in six patients while Aspergillus and Mucor were isolated in two each. The causative organism could not be isolated in eight patients. The size of the ulcer at presentation and height of hypopyon were found to be significant risk factors associated with treatment outcomes. Conclusion: Intrastromal voriconazole as an adjuvant therapy appeared to be effective in treatment of fungal keratomycosis not responding to conventional therapy, thus, reducing the need for therapeutic or tectonic keratoplasty.

Cloning and molecular characterization of a putative habanero pepper SERK1 cDNA expressed during somatic and zygotic embryogenesis

Background: Plant gene homologs that control cell differentiation can be used as biotechnological tools to study the in vitro cell proliferation competence of tissue culture-recalcitrant species such as peppers. It has been demonstrated that SERK1 homologs enhance embryogenic competence when overexpressed in transformed tissues; therefore, cloning of a pepper SERK1 homolog was performed to further evaluate its biotechnological potential. Results: A Capsicum chinense SERK full-length cDNA (CchSERK1) was cloned and characterized at the molecular level. Its deduced amino acid sequence exhibits high identity with sequences annotated as SERK1 and predicted-SERK2 homologs in the genomes of the Capsicum annuum CM-334 and Zunla-1 varieties, respectively, and with SERK1 homologs from members of the Solanaceae family. Transcription of CchSERK1 in plant tissues, measured by quantitative RT-PCR, was higher in stems, flowers, and roots but lower in leaves and floral primordia. During seed development, CchSERK1 was transcribed in all zygotic stages, with higher expression at 14 days post anthesis. During somatic embryogenesis, CchSERK1 was transcribed at all differentiation stages, with a high increment in the heart stage and lower levels at the torpedo/cotyledonal stages. Conclusion: DNA sequence alignments and gene expression patterns suggest that CchSERK1 is the C. chinense SERK1 homolog. Significant levels of CchSERK1 transcripts were found in tissues with cell differentiation activities such as vascular axes and during the development of zygotic and somatic embryos. These results suggest that CchSERK1 might have regulatory functions in cell differentiation and could be used as a biotechnological tool to study the recalcitrance of peppers to proliferate in vitro.

Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

Common bean is a crop recalcitrant to in vitro regeneration and therefore it lacks an efficient transformation protocol that can be reproduced using A. tumefaciens. The main goal of this study was to establish a protocol for A. tumefaciens mediated transformation of Phaseolus vulgaris var. Brunca by marker genes (gusA and nptII) together with the gene for trehalose-6-phosphate synthase from Saccharomyces cerevisiae (TPS1) used in other species to increase tolerance to abiotic stress. The β-glucuronidase activity was detected in 45 % of the LBA4404 ElectroMAX® pCAMBIA1301 infected explants. Transformed explants regenerated new shoots after four to five months period in a kanamycin rich media. Surviving plants were evaluated by PCR and presented an 0.5 % efficiency of transformation. The established protocol for genetic transformation of common bean has two additional advantages with respect to previous reports: (1) it allows for obtaining transformed regenerants and (2) the genetic transformation was stable for the selective gene.

El frijol común en un cultivo recalcitrante a la regeneración in vitro y se carece de un protocolo eficiente y reproducible de transformación genética usando A. tumefaciens. Desarrollamos un protocolo de transformación genética mediada por A. tumefaciens de frijol común variedad Brunca utilizando genes marcadores (gusA y nptII) junto con el gen de la trehalosa-6-fosfato sintasa de levadura (TPS1) utilizado para incrementar tolerancia a estrés abiótico. La actividad de la β-glucoronidasa fue detectada en 45 % de los explantes infectados con la cepa LBA4404 de A. tumefaciens transformada con pCAMBIA1301. Después de 4 o 5 meses se regeneraron tallos en un medio adicionado con kanamicina. Los explantes supervivientes se evaluaron mediante PCR y presentaron una eficiencia de transformación de 0.5 %. El protocolo de transformación genética de frijol común establecido tiene dos ventajas adicionales con respecto a los reportes previos: (1) permite la obtención de regenerares transformados y (2) la transformación genética fue estable para el gen selectivo.

Platelet-Rich Plasma (PRP) Therapy in Dermatology

Platelets present in the human blood produces a wide range of hormones. The activated forms of platelets are actively motile and their motility is clearly shown by the author in a video recorded through a simple microscope and uploaded in YouTube (Moving platelets & PRP_Dr Lokendra Yumnam at www.youtube.com/watch?v=du4nav0Fvy). The platelet-rich plasma can be prepared easily at any side-laboratory attached to the Dermatology OPD. The same may be used in treatment of many skin conditions previously thought to be very difficult to be treated. The author presents results of administering one PRP therapy in patients with androgenic alopecia, disfiguring lesions on face, recalcitrant ulcers including ulcers in leprosy patients, lichen sclerosus, vitiligo and also enhancing skin-graft uptake in scars. The results of PRP therapy in many refractory skin conditions are promising. More researches need to be taken up in this form of magical therapy. This may open up new avenues in the successful treatment of many skin conditions which are hitherto considered to be non-treatable.

Anatomical and biochemical changes associated with thedevelopment and germination of Araucaria angustifolia seeds

This study aimed at assessing the cell cycle, and anatomical and biochemical changes that the embryonic axis of Araucaria angustifolia undergoes during development, focusing on the maturation stage. During all development, cells exhibited intense metabolic activity with an abundance of mitochondria, lipid bodies, and vacuolated cells. The continued accumulation of starch and protein was observed by LM and TEM and indicated by spectra of FTIR. Cell differentiation of the procambium was observed with a thickening of the cell wall and the formation of resiniferous ducts. At Stage III and IV, cells exhibited structural changes such as altered or elongated mitochondria and presence of plastoglobules. These results suggest that there is a gradual transition from developmental metabolism to germination metabolism. Such changes can contribute to the rapid germination of seeds right after their dispersion, making it an ecological strategy to reduce post-dispersal exposure to predators and to avoid damage from reduced moisture.

Rinosinusitis recalcitrante por granulomatosis eosinofílica con poliangeítis: reporte de un caso clínico y revisión de la literatura / Chronic recalcitrant rhinosinusitis secondary to eosinophilic granulomatosis with polyangiitis: report of a case and literature review

RESUMEN Se expone el caso de un paciente de 40 años con diagnóstico de granulomatosis eosinofílica con poliangeítis subyacente a una rinosinusitis crónica recalcitrante. Se describe el caso y se discuten aspectos relevantes de la literatura al respecto.

ABSTRACT We report the case of a 40-year-old man with diagnosis of chronic recalcitrant rhino-sinusitis secondary to eosinophilic granulomatosis with polyangiitis. We described the case and discuss relevant aspects of the literature about it.

Physiological potential of jenipapo seeds stored in different packages / Potencial fisiológico de sementes de jenipapo armazenadas em diferentes embalagens

Seed storage through the reduction of its water content is an important strategy for the conservation of many plant species, but some, such as the jenipapo, are susceptible to desiccation, which may lead to loss of viability and even death, in case its water content is reduced to a critical level. The aim of this work was to evaluate the interference of different packaging conditions in the conservation of the physiological potential of jenipapo seeds stored for 105 days. Specifically, this research sought to analyze the feasibility of the use of vacuum conditions and the coating of the seed with biofilm. The evaluated treatments were: storage in paper bag, plastic bag, plastic with vacuum, plastic with seeds coated with biofilm and plastic with seeds with biofilm and vacuum condition for 15, 45, 75 and 105 days, besides the control. Seeds with biofilm were treated with cassava starch at 3% of the mass to volume ratio. The following tests were performed: water content, germination, germination speed, aerial part and root lengths, wet and dry mass of the aerial part and root. It was concluded that plastic packaging, vacuum condition and biofilm coating are not recommended for storage of jenipapo seeds. Storage in paper bags, when at 24 ºC and with 65% relative humidity for 46 days, is capable of conserving the seeds of jenipapo with germination of 60%.

O armazenamento de sementes por meio da redução do seu teor de água é uma importante estratégia para a conservação de muitas espécies vegetais, porém, algumas, como o jenipapeiro, são sensíveis à dessecação, podendo haver a perda da sua viabilidade, e até a morte, caso o seu teor de água seja reduzido a um nível considerado crítico. Neste trabalho, o objetivo foi avaliar a interferência de diferentes condições de embalagens na conservação do potencial fisiológico de sementes de jenipapo armazenadas por 105 dias. De forma específica, buscou-se analisar a viabilidade do uso de condições de vácuo e do revestimento da semente com biofilme. Os tratamentos avaliados foram: armazenamento em saco de papel, saco plástico, plástico com vácuo, plástico com sementes revestidas com biofilme e plástico com sementes com biofilme e condição de vácuo por 15, 45, 75 e 105 dias, além da testemunha. As sementes com biofilme receberam tratamento com fécula de mandioca a 3% da relação massa por volume. Os seguintes testes foram realizados: teor de água, germinação, velocidade de germinação, comprimento da parte aérea e radicular, massa úmida e seca da parte aérea e da raiz. Concluiu-se que a embalagem plástica, a condição de vácuo e o revestimento com biofilme não são recomendados para o armazenamento de sementes de jenipapo. O armazenamento em saco de papel quando realizado em ambiente com 24 ºC e 65% de umidade relativa do ar durante 46 dias é capaz de conservar as sementes de jenipapo com germinação de 60%.

Aplicação de ozônio e de ozônio + peróxido de hidrogênio para remoção de compostos recalcitrantes em lixiviados de aterros sanitários / Application of ozone and ozone + hydrogen peroxide to remove recalcitrant compounds in landfill leachates

RESUMO Os lixiviados são efluentes gerados em depósitos de resíduos sólidos por meio da decomposição da matéria orgânica, somados às águas de infiltração e a materiais suspensos. Possuem compostos biodegradáveis e recalcitrantes, de difícil degradação biológica (biodegradação). Esse tipo de efluente necessita de um tratamento antes de ser disposto no meio ambiente. O objetivo deste trabalho foi avaliar o uso de ozônio (O3) e de peróxido de hidrogênio (H2O2) como processo oxidativo avançado (POA), visando à remoção de compostos recalcitrantes em lixiviados de aterros sanitários. Foram utilizados dois lixiviados provenientes do aterro sanitário de São Leopoldo, Rio Grande do Sul: o bruto e o efluente das lagoas de tratamento, os quais foram tratados, nesta pesquisa, apenas com O3 em concentração de 29 mg.L-1 (gerados a partir de processo corona) ou com adição de 1.000 mg.L-1 de H2O2 ao O3. Os ensaios, em escala piloto (reator de 460 L de volume útil), tiveram a duração de 96 horas. O POA deste estudo mostrou-se mais eficiente no tratamento de lixiviados com as menores concentrações iniciais de demanda bioquímica de oxigênio (DBO), demanda química de oxigênio (DQO) e carbono orgânico total (COT), independente de ser lixiviado bruto ou tratado. Altas remoções de cor aparente, DQO e COT foram observadas, chegando a 99, 88 e 66%, respectivamente. A maior remoção de carbono inorgânico (CI) foi 23%, porém o grupo que iniciou com a maior concentração desse parâmetro obteve as maiores remoções para todos os outros critérios estudados. A adição de H2O2 aumentou a eficiência de remoção de todos os fatores analisados, porém não de forma significativa. Observou-se necessidade de tratamento complementar ao POA deste estudo.

ABSTRACT Leachates are wastewaters generated at sites of solid waste disposal through organic matter decomposition, added to infiltration waters and suspended materials. They contain biodegradable and recalcitrant compounds, difficult to biological degradation (biodegradation). This wastewater requires treatment before being disposed in the environment. The aim of this study was to evaluate the use of ozone (O3) and hydrogen peroxide (H2O2) as advanced oxidation process (AOP), in order to remove recalcitrant compounds in landfill leachate. Two leachates from São Leopoldo landfill, in Rio Grande do Sul, were used: raw and treated in pounds, which were treated only with O3 at the concentration of 29 mg.L-1 (generated from the corona process) or with addition of 1,000 mg.L-1 H2O2 to O3. The tests occurred on a pilot scale, treating 460 L of leachate in each test, which lasted 96 hours. AOP of this study was more efficient in the treatment of leachate that had the lowest initial concentrations of biological oxygen demand (BOD), chemical oxygen demand (COD) and total organic carbon (TOC) (both raw and treated leachate) for both of the leachates. High removals of apparent color, COD and TOC were observed, reaching 99, 88 and 66%, respectively. The greater removal of inorganic carbon was 23%, but the group that started with the highest concentration of this parameter achieves the greatest removals for all other parameters studied. The addition of H2O2 increased the efficiency of removal of all parameters, but not significantly. It is necessary additional treatment to the AOP of this study.

Clinical study of ultrasound - guided injection of platelet-rich plasma for the treatment of recalcitrant lateral epicondylitis / 介入放射学杂志

Objective To explore the clinical efficacy of ultrasound - guided injection of platelet-rich plasma (PRP) for the treatment of recalcitrant lateral epicondylitis. Methods From September 2014 to June 2016, a total of 15 patients with recalcitrant lateral epicondylitis received ultrasound - guided injection of autologous PRP therapy, including via left arm injection (n=2) and via right arm injection (n=13). By using twice centrifugal method, the patient' s own venous whole blood was centrifuged to obtain PRP. All patients underwent PRP injection once a week, a total of 3 treatments were performed for each patient. Results After the first injection of PRP, the patients were followed up for 12 months. One month after the treatment, visual analogue scale (VAS) score was obviously improved, at 3 months after the treatment the improvement of VAS score reached its peak and it remained at this level until 12 months after the treatment. The elbow joint function, which was evaluated with modified MAYO elbow score, was also significantly improved in one month after the treatment, and the clinical effect was sustained to 12 months after the treatment. Conclusion Ultrasound - guided precise injection of PRP can effectively improve the pain and the elbow joint function caused by recalcitrant lateral epicondylitis. (J Intervent Radiol, 2018, 27:238-241)

Three Cases of Generalized Granuloma Annulare Successfully Treated with Methotrexate / 대한피부과학회지

Granuloma annulare is a benign granulomatous skin disease, clinically characterized by dermal papules tending to form annular shapes. Granuloma annulare is classified into localized, generalized, subcutaneous, and perforating types. Localized granuloma annulare is likely to resolve spontaneously, while generalized granuloma annulare usually takes a chronic course. Topical steroids, phototherapy, isotretinoin, dapsone, hydroxychloroquine, and other various treatments have been proposed, but some cases have been resistant to all of those treatment modalities. We experienced 3 cases of generalized granuloma annulare demonstrating a recalcitrant course that were successfully treated with methotrexate and propose that methotrexate may be an effective option for recalcitrant generalized granuloma annulare treatment.

Pathogenesis of Recalcitrant Chronic Rhinosinusitis: The Emerging Role of Innate Immune Cells

Chronic rhinosinusitis (CRS) is a major part of the recalcitrant inflammatory diseases of the upper airway that needs enormous socioeconomic burden. T helper (Th) 2 type immune responses recruiting eosinophils were the most well-known immune players in CRS pathogenesis especially in western countries. By the piling up of a vast amount of researches to elucidate the pathogenic mechanism of CRS recently, heterogeneous inflammatory processes were found to be related to the phenotypes of CRS. Recently more cells other than T cells were in the focus of CRS pathogenesis, such as the epithelial cell, macrophage, innate lymphoid cells, and neutrophils. Here, we reviewed the recent research focusing on the innate immune cells related to CRS pathogenesis.

Behavior of lateral buds of Hancornia speciosa after cryopreservation by encapsulation-vitrification / Comportamento de gemas laterais de Hancornia speciosa após criopreservação por encapsulamento-vitrificação

Hancornia speciosa is a fruitful species from Cerrado biome with high economic potential. However, the intense and disordered extractivism have caused a reduction of its population in its endemic area. In addition, seed recalcitrance negatively affects the conventional conservation of the species. Aiming to find alternatives that enable the long-term conservation of this species, the study's objective was to assess the behavior of lateral bud's regrowth after cryopreservation procedures by encapsulation-vitrification technique. Sodium alginate capsules containing lateral buds were pre-cultured in liquid WPM supplemented with 1.0 M glycerol, and subsequently exposed to different concentrations of sucrose (0.3; 0.75 and 1.0 M) for 24 or 48 hours. The capsules were subjected to dehydration in silica gel or airflow hood for 0, 1, 2 and 3 hours before different incubation times in PVS2 (0, 15, 30, 60 and 120 minutes) at 0°C. A high regeneration percentage of lateral buds was observed after cryopreservation of capsules treated with 0.75 M sucrose plus 1.0 M glycerol (24 hours), associated with dehydration in an airflow hood (1 hour) and immersion in PVS2 (15 minutes). Encapsulation-vitrification allowed the long-term conservation, and provided high plant material survival rates after cryopreservation of Hancornia speciosa sensitive explants.

Hancornia speciosa é uma frutífera do Cerrado com elevado potencial econômico. Entretanto, o extrativismo desordenado causou a redução populacional em sua área endêmica. Além disso, a recalcitrância da semente afeta negativamente sua conservação convencional. Buscando alternativas de conservação para essa espécie, objetivou-se avaliar a regeneração das gemas laterais após a técnica de encapsulamento-vitrificação. Cápsulas de alginato de sódio contendo gemas laterais foram pré-cultivadas em meio WPM acrescido de 1,0 M de glicerol e, posteriormente, imersas em diferentes concentrações de sacarose (0,3; 0,75 e 1,0 M) por 24 ou 48 horas. As cápsulas foram submetidas à desidratação em sílica gel ou em fluxo laminar por 0, 1, 2 e 3 horas antes de sua incubação em diferentes tempos de PVS2 (0, 15, 30, 60 e 120 minutos). Elevada porcentagem de regeneração de gemas laterais foi observada após a criopreservação de cápsulas tratadas com 0,75 M de sacarose + 1,0 M de glicerol por 24 horas, associado com a desidratação em fluxo laminar (1 hora) e imersão em PVS2 (15 minutos). O encapsulamento-vitrificação é eficiente para a conservação de longo prazo e permite a obtençao de altas taxas de sobrevivência após a criopreservação de explantes sensíveis (gemas laterais) de Hancornia speciosa.

Low-Dose Systemic Methotrexate Therapy for Recalcitrant Alopecia Areata

BACKGROUND: Alopecia areata (AA) is an autoimmune skin disease difficult to manage and treat. The pathogenesis of AA features a T-cell-associated autoimmune process, and systemic immunosuppressive therapy is prescribed widely for AA. OBJECTIVE: To evaluate the efficacy and tolerance of systemic low-dose methotrexate (LD-MTX) therapy in treatment of recalcitrant AA multiplex. METHODS: In a retrospective, non-controlled study, we evaluated 29 patients with recalcitrant AA treated with LD-MTX and assessed the therapeutic response according to severity of disease, disease duration, cumulative dose of MTX, and drug safety. RESULTS: MTX was administered twice weekly, and the mean maximum weekly dose was 14.48 mg. The response was A5 (regrowth=100.0%) in 14 (48.3%) patients and A4 (regrowth of 75%~90%) in 12 (41.4%) patients. Three patients had poor response to LD-MTX treatment (A2: n=2 [6.9%], A1: n=1 [3.4%]). All three of the patients showing a poor response had disease durations exceeding 24 months. Relapse was observed in 31% of patients with more than 75% regrowth. Common side-effects were elevated liver enzyme levels and gastrointestinal discomfort. CONCLUSION: LD-MTX appears to be an effective and well-tolerated treatment for recalcitrant AA multiplex.

Storage on the vigor and viability of macauba seeds from two provenances of Minas Gerais State / Tipo de armazenamento no vigor e na viabilidade de sementes de macaúba oriundas de duas localidades do Estado de Minas Gerais

ABSTRACT: Macauba palm stands out for having favorable features to biodiesel production such as the high oil content of its fruit. Considering the great potential of the species and their applicability in the renewable energy field, it becomes indispensable to establish the right conditions for storing the seeds for propagation purpose. The aim of this research was to evaluate the effect of seed moisture content, packaging, and storage conditions such as temperature and relative humidity on the quality of seeds from Minas Gerais State, during a 12-month storage period. The research had two independent assays: (I) the seeds were stored with three moisture contents/ranges 4.0≤6.0%; 6.0≤8.0% and 8.0≤10.0% in impermeable packages, under room temperature and at 10ºC; (II) seeds with approximately 5.9% of moisture content were stored in three different types of packages: a) permeable, b) semi-permeable and c) impermeable. Three storing conditions were tested: a) room temperature and RH under laboratory conditions; b) 15ºC and 45% RH; c) 20ºC and 55% RH. Water content, germination rate and germination speed index were evaluated at 0, 4, 8 and 12 months of storing. The best germination results were obtained with the moisture range of 6.0≤8.0%, with seeds kept at room temperature; while the seeds stored at 10ºC, regardless the moisture range, did not survive. The stored seeds with 5.9% moisture content and at both 15ºC/45%RH and 20ºC/55% RH conditions, independently of the package type used, showed the best results. Thus, macaw palm seeds can be classified as intermediates seeds.

RESUMO: Considerando o grande potencial oleaginoso da macauba e a sua aplicabilidade nos setores energéticos e industriais, torna-se indispensável o conhecimento das condições adequadas de armazenamento das sementes, garantindo a manutenção da viabilidade. O objetivo do trabalho foi avaliar o efeito do teor de água da semente e das condições de armazenamento (temperatura, umidade relativa do ar e embalagem) sobre a qualidade das sementes, oriundas de Minas Gerais, durante 12 meses. O experimento foi realizado através de dois ensaios: no primeiro, as sementes foram armazenadas sob três faixas de umidade, 4,0≤6,0%; 6,0≤8,0% e 8,0≤10,0%, em embalagens impermeáveis, em temperatura ambiente e a 10°C. No segundo, as sementes foram acondicionadas em três tipos de embalagem: a) permeável; b) semipermeável; e c) impermeável. Foram comparadas três condições de armazenamento: a) temperatura e UR ambientes, em condição de laboratório; b) 15ºC e 45% UR; c) 20ºC e 55% UR. Avaliou-se o teor de água, a taxa de germinação e o índice de velocidade de germinação aos 0, 4, 8 e 12 meses de armazenamento. A faixa de umidade de 6,0≤8,0% em temperatura ambiente possibilitou a melhor conservação das sementes durante os 12 meses de armazenamento. Porém, as sementes não sobreviveram em temperatura de 10°C, independente dos teores de umidade. O teor de água de 5,9% e os ambientes de armazenamento de 15°C/45% UR e 20°C/55% UR, independente do tipo de embalagem, apresentaram os melhores resultados ao longo do armazenamento. Dessa forma, as sementes de macauba podem ser classificadas como sementes intermediárias.

Early harvest increases post-harvest physiological quality of Araucaria angustifolia (Araucariaceae) seeds

ResumenAraucaria angustifolia es una conífera nativa de Brasil y una especie en peligro de extinción. Sus semillas tienen un corto período de viabilidad, factor que contribuye a su vulnerabilidad. Este estudio tuvo como objetivo evaluar la calidad fisiológica durante el período de desarrollo y post-almacenamiento de semillas de A. angustifolia. Conos de A. angustifolia fueron recolectados en poblaciones naturales en Curitibanos, Santa Catarina, Brasil, en marzo, abril, mayo y junio y clasificados en los estadios de desarrollo cotiledonar i, ii y iii de acuerdo con el mes de recolecta. Un total de 10 conos fueron recolectados para cada estadio. Las semillas fueron almacenadas en refrigerador durante 60 y 120 días y posteriormente sometidas a pruebas de germinación (25 °C - fotoperiodo de 12 h) siendo evaluados el contenido de humedad, tetrazolio y el vigor (conductividad eléctrica [75 mL de agua destilada a 25 °C], índice de velocidad de germinación, y la longitud de la parte aérea y de la raíz). Durante el desarrollo de las semillas, el contenido de humedad se redujo desde el estadio cotiledonar (66.54 %) al estadio iii (47.44%), y el vigor aumentaron en el último estadio. Durante el almacenamiento, el contenido de humedad en el estadio cotiledonar y estadio i fue estable. Entretanto, las semillas almacenadas mostraron una reducción en el contenido de humedad después de 120 días en los estadios ii y iii. La calidad fisiológica en el estadio cotiledonar mostró un aumento de 86 % y 93 % de germinación después de 60 y 120 días de almacenamiento, respectivamente, a diferencia de los estadios ii y iii, los cuales mostraron una disminución en la viabilidad de las semillas y en el vigor después del almacenamiento. La conductividad eléctrica fue mayor para las semillas en estadio cotiledonar recién recolectadas que para aquellas almacenadas durante 60 y 120 días. Sin embargo, en otras estadios, el contenido de lixiviados después de 120 días de almacenamiento aumentó con el avance del período de recolecta. El índice de velocidad de germinación y la longitud de la parte aérea y raíz después del almacenamiento eran más altos para las semillas en el estadio cotiledonar y el estadio i, a diferencia del estadio ii y iii, los cuales tenían raíz y parte aérea de menor longitud durante el almacenamiento. Por lo tanto, el mantenimiento del contenido de humedad de la semilla durante el almacenamiento fue variable y depende del período de recolecta. Además, la calidad fisiológica de las semillas difiere entre los estadios más tempranos o tardíos. Una recolección precoz favoreció la calidad fisiológica de las semillas, y puede ser una estrategia para aumentar la conservación de semillas de A. angustifolia.

AbstractAraucaria angustifolia is a conifer native to Brazil and is an endangered species. Since this species seeds have a short period of viability, its vulnerability is higher. Thus the aim of this study was to evaluate the physiological quality of A. angustifolia seeds during the development and post-storage periods. For this, cones of A. angustifolia were collected from a natural population in Curitibanos, Santa Catarina, Brazil, in March, April, May and June 2012. The collected seeds were classified into developmental stages of cotyledonary, i, ii and iii according to the month of collection; a total of 10 cones were collected for each stage. Seeds were stored in a refrigerator for 60 and 120 days, and were submitted to a chamber germination test (25 °C-photoperiod 12 h). Additionally, seeds were tested for moisture content (105 °C for 24 hours), tetrazolium (0.1 % for 1 hour) and vigor (electric conductivity [75 mL distilled water at 25 °C], germination speed index, and shoot and root length). Our results showed that during seed development, moisture content decreased from the cotyledonary stage (66.54 %) to stage iii (49.69 %), and vigor increased in the last stage. During storage, moisture content at cotyledonary stage and stage i was stable. On the other hand, stored seeds exhibited a decrease in moisture content after 120 days at stages ii and iii. Physiological quality at the cotyledonary stage resulted in an increased germination rate of 86 % and 93 % after 60 and 120 days of storage, respectively; unlike stages ii and iii exhibited a decrease in seed viability and vigor after storage. Electrical conductivity was higher for fresh seeds at the cotyledonary stage, than for those stored for 60 and 120 days. However, in other stages, released leachate content after 120 days of storage, increased with the advance of the collection period. Germination speed index and shoot and root lengths after storage were highest for seeds at the cotyledonary stage and stage i; unlike stages ii and iii which had short root and shoot lengths during storage. Thus, the maintenance of seed moisture content during storage was variable and dependent on the period of collection. Furthermore, the physiological quality differed among earlier and later stages. Early collection favored seed physiological quality, and may be a strategy for better conservation of A. angustifolia seeds. Rev. Biol. Trop. 64 (2): 885-896. Epub 2016 June 01.

A Clinical Study using Quadrivalent Human Papilloma Virus (HPV) Vaccine for Treatment of Recalcitrant Wart / 대한피부과학회지

BACKGROUND: Although several traditional treatments have been applied for recalcitrant viral warts, these treatments have rarely resulted in complete recovery. To treat the recalcitrant viral wart, alternative therapies are required. OBJECTIVE: This study aimed to evaluate the efficacy and safety of quadrivalent HPV vaccine for recalcitrant wart treatment. METHODS: From 2012 to 2014, 17 patients who provided informed consent were enrolled. All patients received 3 doses of quadrivalent HPV vaccine at 0, 2, and 6 months, respectively. During clinic visits, doctors checked the grade of improvement, patient satisfaction, and treatment side effects. After completion of the 3 doses, the patients were followed up for 5 months with outpatient visits and telephone inquiries. RESULTS: After the third dose, 58.8% of patients showed complete remission and 41.2% showed no response after 5 months. There were no statistically significant differences in sex, age, disease duration, number, anatomic site, and previous treatment between the complete remission group and the no-response group. An adverse effect (syncope) was observed in one patient. CONCLUSION: Compared with traditional aggressive therapies, quadrivalent HPV vaccine is a simple method and does not usually interfere with the patient's work or social life. Quadrivalent HPV vaccine is an effective and safe alternative treatment for recalcitrant warts.