Abstract Rusts and powdery mildews pose serious threats to wheat and have caused substantial yield losses worldwide. Host resistance is the most economical and sustainable approach for managing such diseases. In this study, an effective leaf rust resistance gene Lr45-derived from Secale cereale L. and a linked stem rust and powdery mildew resistance gene Sr36/Pm6-derived from Triticum timopheevii were successfully pyramided. They were validated into well-adapted Indian wheat cultivars that were already carrying the APR stem rust gene Sr2/Lr27/Yr30 through marker-assisted backcross selection (MABC) following two parallel backcrossing schemes. Three efficiently linked microsatellite markers, G372185 (Lr45), Stm773-2(Sr36), and Xgwm533 (Sr2), were used to confirm introgression of these genes. Lines with resistance genes in each background showed improved agronomic traits in comparison to their recurrent parents. These lines could be used in wheat improvement programs as potentially resistant stocks for leaf, stem rusts and powdery mildew to develop new wheat cultivars.

ABSTRACT Background: Rapid molecular methods such as the line probe assay (LPA) and Xpert® MTB/RIF assay (Xpert) have been recommended by the World Health Organization for drug-resistant tuberculosis (DR-TB) diagnosis. We conducted an interventional trial in DR-TB reference centers in Brazil to evaluate the impact of the use of LPA and Xpert. Methods: Patients with DR-TB were eligible if their drug susceptibility testing results were available to the treating physician at the time of consultation. The standard reference MGITTM 960 was compared with Xpert (arm 1) and LPA (arm 2). Effectiveness was considered as the start of the appropriate TB regimen that matched drug susceptibility testing (DST) and the proportions of culture conversion and favorable treatment outcomes after 6 months. Results: A higher rate of empirical treatment was observed with MGIT alone than with the Xpert assay (97.0% vs. 45.0%) and LPA (98.2% vs. 67.5%). Patients started appropriate TB treatment more quickly than those in the MGIT group (median 15.0 vs. 40.5 days; p<0.01) in arm 1. Compared to the MGIT group, culture conversion after 6 months was higher for Xpert in arm 1 (90.9% vs. 79.3%, p=0.39) and LPA in arm 2 (80.0% vs. 83.0%, p=0.81). Conclusions: In the Xpert arm, there was a significant reduction in days to the start of appropriate anti-TB treatment and a trend towards greater culture conversion in the sixth month.

Introduction Carotid artery injury (CAI) is the most feared and potentially catastrophic intraoperative complication an endoscopic skull base surgeon may face. With the advancement of transnasal endoscopic surgery and the willingness to tackle more diverse pathology, evidence-based management of this life-threatening complication is paramount for patient safety and surgeon confidence. Objectives We review the current English literature surrounding the management of CAI during endoscopic transnasal surgery. Data Synthesis The searched databases included PubMed, MEDLINE, Cochrane database, LILACS, and BIREME. Keywords included “sinus surgery,” “carotid injury,” “endoscopic skull base surgery,” “hemostasis,” “transsphenoidal” and “pseudoaneurysm.” Conclusions Review of the literature found the incidence of CAI in endonasal skull base surgery to be as high as 9% in some surgeries. Furthermore, current treatment recommendations can result in damage to critical neurovascular structures. Management decisions must be made in the preoperative, operative, and postoperative setting to ensure adequate treatment of CAI and the prevention of its complications such as pseudoaneurysm. Emphasis should be placed on surgical competency, teamwork, and technical expertise through education and training.

A abordagem cirúrgica do saco lacrimal por via intranasal assistida por videoendoscopia é, hoje, realizada com altos índices de sucesso. Apesar da técnica tradicional por via externa apresentar resultados bastante satisfatórios, tem a grande desvantagem de necessitar de uma incisão de pele e sua consequente cicatriz local. Com o desenvolvimento e aprimoramento do instrumental e das técnicas endonasais, a abordagem endoscópica é cada vez mais utilizada. OBJETIVO: Este artigo revisa a anatomia do sistema lacrimal, a avaliação pré-operatória e os detalhes técnicos da cirurgia assistida por endoscopia que podem proporcionar os desfechos cirúrgicos mais favoráveis ao paciente. As complicações e as causas de insucesso cirúrgico são também brevemente revisadas. METODOLOGIA: Trata-se de uma revisão da experiência dos autores nos últimos 10 anos de aplicação da técnica endoscópica para cirurgia do saco lacrimal. CONCLUSÃO: Os resultados da dacriocistorrinostomia endoscópica são, no mínimo, iguais aos da técnica tradicional externa. Apesar disto, o trabalho conjunto do oftalmologista e do otorrinolaringologista é muito vantajoso para o melhor manejo possível do paciente com epífora.

The endonasal surgical approach of the lacrymal sac assisted by video-endoscopy is carried out today with high success rates. Despite the satisfactory results reached with the traditional external approach, it has the disadvantage of requiring a skin incision and a consequent local scar. With the development and enhancement of the endonasal techniques, the endoscopic approach is increasingly preferred by surgeons. OBJECTIVE: This paper reviews the lacrymal system anatomy, the preoperative assessment and the technical details of the endoscopic assisted approach which may provide better surgical outcomes for patients. We will also briefly discuss complications and causes for surgical failure. METHODOLOGY: This is a review of the experience of the authors in the past 10 years of employing the endoscopic technique for the lacrymal sac surgery. CONCLUSION: Outcomes regarding the endoscopic dacryocystorhinostomy are, at leas, equal to those from the traditional external approach. Notwithstanding, the joint work between the otorhinolaryngologist and the ophthalmologist is of great benefit to patients with epiphora.

OBJETIVO: Avaliar a viabilidade celular de S. aureus in vitro após a exposição de riboflavina (B2) e luz ultravioleta A (UVA). MÉTODOS: Amostras de S. aureus colocadas em uma placa de 96 poços (em triplicata) foram expostas a riboflavina 0,1% (B2) e luz ultravioleta (comprimento de onda de 365 nm) poder de 3 mW/cm², 8 mm de diâmetro, por 30 minutos. Grupos controles foram também preparados em triplicata: controle branco, somente luz ultravioleta A, somente riboflavina e controle morto. A viabilidade bacteriana foi analisada usando microscópio de fluorescência. Para investigar a ocorrência de micro-organismos "viáveis porem não cultiváveis" a viabilidade celular foi avaliada utilizando-se placas de meio de cultivo bacteriano. Analise estatística foi realizada utilizando-se os valores obtidos em triplicata de cada grupo experimental. RESULTADOS: Nenhuma diferença foi observada entre o grupo tratamento e os grupos controle (p=1). CONCLUSÃO: A combinação riboflavina 0,1% e luz ultravioleta 365 nm de comprimento de onda não demonstrou atividade antimicrobiana contra S. aureus oxacilina sensível.

PURPOSE: To assess S. aureus in vitro viability after the exposure to ultraviolet light A (UVA) and riboflavin (B2). METHODS: Samples of S. aureus in 96 well plates (in triplicate) were exposed to riboflavin (B2) and ultraviolet light A (365 nm wavelength) at a power density of 3 mW/cm², 8 mm spot diameter, for 30 minutes. Control groups were prepared as well in triplicate: blank control, ultraviolet light A only, riboflavin only and dead bacteria Control. The bacterial viability was measured using fluorescent microscopy. In order to investigate the occurrence of "viable but non-culturable" microorganisms after treatment, the cell viability was also investigated by plate culture procedure onto a broth medium. Statistical analysis was performed using the triplicate values from each experimental condition. RESULTS: No difference was observed among the treatment group and the control samples (p=1). CONCLUSION: The combination of riboflavin 0.1% and ultraviolet light A at 365 nm did not exhibit antimicrobial activity against oxacillin susceptible S. aureus.

Lisina, treonina, metionina e isoleucina são derivados do aspartato. Aspartato quinase tem um papel chave no controle da via metabólica do aspartato. A enzima é muito sensível à manipulação e armazenagem e o ensaio enzimático do hidroxamato que é normalmente utilizado para determinar a atividade da aspartato quinase deve ser alterado de acordo com a espécie de planta e tecido vegetal a ser analisado. Nós otimizamos o ensaio para a determinação da atividade da aspartato quinase em culturas celulares de calos de milho. Entre os vários parâmetros testados do ensaio, a concentração de ATP/Mg e temperatura foram críticos para atividade enzimática. No caso da temperatura, 35°C foi a temperatura ótima para atividade da aspartato quinase.

Lysine, threonine, methionine and isoleucine are synthesized from aspartate in a branched pathway in higher plants. Aspartate kinase plays a key role in the control of the aspartate pathway. The enzyme is very sensitive to manipulation and storage and the hydroxamate assay normally used to determine aspartate kinase activity has to be altered according to the plant species and tissue to be analyzed. We have optimized the assay for the determination of aspartate kinase in maize plants callus cell cultures. Among all the assay parameters tested, the concentration of ATP/Mg and temperature were critical for enzyme activity. In the case of temperature, 35°C was shown to be the optimum temperature for aspartate kinase activity.

A contaminação do ambiente por metais tóxicos torna-se uma ameaça tanto para o homem quanto para a biosfera, impedindo o desenvolvimento da agricultura e prejudicando os ecossistemas. Em nações desenvolvidas, esse problema está sendo direcionado e resolvido, consideravelmente, com o uso de plantas tolerantes especializadas na limpeza de substratos contaminados e poluídos com metais. O uso de plantas que naturalmente acumulam metais e a aplicação da engenharia genética acelerariam o processo de transferência de toda essa tecnologia do laboratório ao campo. É essencial, portanto, investigar e entender como essas plantas funcionam e quais os caminhos metabólicos e genes envolvidos no processo. A combinação das tecnologias das "ômicas" tem um prodigioso potencial no desenvolvimento dessa tecnologia verde. Entretanto, estratégias para produzir plantas geneticamente alteradas para remoção, destruição ou seqüestro de substâncias tóxicas do ambiente e suas implicações devem ser cuidadosamente investigadas.

The contamination of the environment by toxic metals poses a threat for "Man and biosphere", reducing agricultural productivity and damaging the health of the ecosystem. In developed nations, this problem is being addressed and solved to some extent by using "green technology" involving metal tolerant plants, to clean up the polluted soils. The use of naturally occurring metal tolerant plants and the application of genetic manipulation, should hasten the process of transferring this technology from laboratory to field. Therefore, it is essential to investigate and understand how plants are able to tolerate toxic metals and to identify which metabolic pathways and genes are involved in such a process. Recent advances in knowledge derived from the "omics", have considerable potential in developing this green technology. However, strategies to produce genetically altered plants to remove, destroy or sequester toxic metals from the environment and the long-term implications, must be investigated carefully.

As sementes dos cereais são pobres em aminoácidos essenciais, principalmente lisina, triptofano e treonina. Os aminoácidos lisina e treonina são sintetizados na via metabólica do aspartato. Apesar de a maioria das enzimas da via do aspartato ter sido isolada e caracterizada em várias espécies de plantas, o metabolismo de lisina e treonina é totalmente desconhecido em sorgo. Isolamos duas enzimas, aspartato quinase (AK) e homosserina desidrogenase (HSDH) de sorgo, estabelecendo condições ótimas de ensaio para determinação da atividade de AK e HSDH. Observaram-se as atividades mais elevadas de AK e HSDH em sementes imaturas. A atividade de AK foi inibida por treonina e lisina, indicando a existência de duas isoenzimas, uma sensível à inibição por treonina e, a outra, sensível à inibição pela lisina, predominando, esta última, nas sementes de sorgo. A atividade de HSDH foi inibida por treonina, indicando a existência de uma isoenzima sensível à treonina, a maioria da atividade, entretanto, não foi inibida por treonina, sugerindo a existência de uma segunda isoenzima de HSDH predominante resistente à inibição pela treonina.

Cereal seeds are poor in essential amino acids, particularly lysine, tryptophan and threonine. The amino acids lysine and threonine are synthesized in the aspartate pathway. Although most of the enzymes of the aspartate pathway have been isolated and characterized in higher plant species, the metabolism of lysine and threonine is totally unknown in sorghum. We have isolated two enzymes, aspartate kinase (AK) and homoserine dehydrogenase (HSDH) from sorghum. Optimum assay conditions were established for the determination of AK and HSDH activities. The highest level of activity was observed in immature seeds. AK was shown to be inhibited by threonine and lysine indicating the existence of at least two isoenzymes, one sensitive to threonine inhibition and the other sensitive to lysine inhibition with the latter being predominant in sorghum seeds. HSDH was shown to be inhibited by threonine indicating the existence of a threonine-sensitive HSDH, however, most of the activity was not inhibited by threonine, suggesting the existence of a second predominant isoenzyme of HSDH resistant to threonine inhibition.

A principal fonte de proteínas vegetais para alimentação humana e animal é fornecida pelos grãos de cereais e leguminosas. O conteúdo de aminoácidos solúveis totais foi determinado por HPLC em endospermas de milho (Zea mays L.) normal e mutantes opaco e floury. A concentração de aminoácidos solúveis totais variou entre os mutantes. Os mutantes o11 e o13 se destacaram com médias superiores, enquanto que mutantes o10, fl3 e fl1 apresentaram as menores médias. De maneira geral, para a concentração total de aminoácidos solúveis, os grãos dos mutantes foram similares aos seus tipos selvagens com exceção dos mutantes o11 e fl1 sendo que o primeiro apresentou valor maior que seu tipo selvagem W22, enquanto que o fl1 teve valor menor que o Oh43. Para metionina, os mutantes o2 e o11 e o tipo selvagem Oh43 apresentaram as mais altas concentrações deste aminoácido. Valores similares foram observados entre os mutantes e os tipos selvagens para concentração de outros aminoácidos analisados, tais como lisina e treonina. As altas concentrações sugeridas originalmente para estes mutantes devem ser devidas aos níveis destes aminoácidos incorporados nas proteínas de reserva, mas não na forma solúvel.

For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperm of wild-type, opaque and floury maize (Zea mays L.) mutants were determined by HPLC. The total absolute concentration of soluble amino acids among the mutants varied depending on the mutant. The o11 and o13 mutants exhibited the highest average content, whereas o10, fl3 and fl1 exhibited the lowest average content. In general, the mutants exhibited similar concentrations of total soluble amino acids when compared to the wild-type lines, with the clear exception of mutants o11 and fl1, with the o11 mutant exhibiting a higher concentration of total soluble amino acids when compared to its wild-type counterpart W22 and the fl1 mutant a lower concentration when compared to its wild-type counterpart Oh43. For methionine, the mutants o2 and o11 and wild-type Oh43 exhibited the highest concentrations of this amino acid. Significant differences were not observed between mutants for other amino acids such as lysine and threonine. The high lysine concentrations obtained originally for these mutants may be due to the amino acids incorporated into storage proteins, but not those present in the soluble form.

A lisina é um aminoácido essencial cuja via de biossíntese faz parte da via metabólica do ácido aspártico, pela qual são também sintetizados os aminoácidos treonina, metionina e isoleucina. Além disso, a lisina é o principal aminoácido limitante em todos os cereais e por cerca de 30 anos a via do ácido aspártico tem sido estudada em plantas, com o intuito de desvendar e caracterizar os principais pontos chave na regulação das vias de biossíntese desses aminoácidos. Duas etapas distintas, uma primeira originada a partir do desenvolvimento da cultura de tecidos (anos 70-80) e a segunda a partir do desenvolvimento de técnicas para a transformação de plantas (anos 90), permitiram que mutantes bioquímicos e plantas trangênicas fossem produzidos com alterações específicas em passos metabólicos chave, levando à superprodução e acúmulo de treonina em vários tecidos das plantas. Entretanto, a acumulação de lisina em sementes não foi obtida. Tal fato, associado a estudos bioquímicos da via de degradação da lisina em cereais e em leguminosas, indicou que a manipulação da degradação seria tão ou mais importante que a manipulação da biossíntese de lisina para o acúmulo deste aminoácido em sementes dos cereais . Em milho, o uso e estudo de outros mutantes tais como o opaco-2 e variedades QPM (Quality Protein Maize) contribuíram significativamente para a compreensão dos eventos regulatórios. As estratégias para a obtenção de materiais ricos em lisina e sua relevância à manipulação de outros aminoácidos são revisados.

The nutrition value of a protein is directly related to its amino acid composition. Some of these amino acids, termed essential amino acids, cannot be synthesized by humans and therefore must be supplied in the diet for adults and in particular for infants and children. Lysine is an essential amino acid synthesized via the aspartic acid metabolic pathway, in which threonine, methionine and isoleucine are also endproducts. Moreover, lysine is the first limiting amino acid in all cereal grains. For over 30 years, the aspartic acid metabolic pathway has been studied in higher plants with the aim of identifying and characterizing the key regulatory points controlling the biosynthetic pathway. Two clear distinct time periods, one begining with the development of tissue culture techniques (1970-80's) and the second with the development of plant transformation techniques (90's), has encouraged the production of biochemical mutants and transgenic plants with specific alterations in key enzymes of the pathway, leading to the overproduction and accumulation of threonine in all plant tissues. However, the accumulation of lysine in seeds has been particularly difficult to achieve. Such an observation, associated with the recent biochemical studies on lysine degradation in cereal and legume plant species, has indicated that the manipulation of lysine degradation is as important as the manipulation of lysine synthesis, if the goal of accumulating this amino acid in cereal seeds is to be achieved. In maize, the study and use of other mutants such as the opaque-2 and QPM (Quality Protein Maize) varieties, has contributed significantly to our understanding of the regulatory aspects of the aspartate pathway. The strategies of obtaining cereals rich in lysine and their relevance to the manipulation of other amino acids have been discussed.