Overexpression of NtAGPase small subunit gene increases leaf starch content and tobacco biomass / 生物工程学报

Production of biofuels such as ethanol from non-grain crops may contribute to alleviating the global energy crisis and reducing the potential threat to food security. Tobacco (Nicotiana tabacum) is a commercial crop with high biomass yield. Breeding of starch-rich tobacco plants may provide alternative raw materials for the production of fuel ethanol. We cloned the small subunit gene NtSSU of ADP-glucose pyrophosphorylase (NtAGPase), which controls starch biosynthesis in tobacco, and constructed a plant expression vector pCAMBIA1303-NtSSU. The NtSSU gene was overexpressed in tobacco upon Agrobacterium-mediated leaf disc transformation. Phenotypic analysis showed that overexpression of NtSSU gene promoted the accumulation of starch in tobacco leaves, and the content of starch in tobacco leaves increased from 17.5% to 41.7%. The growth rate and biomass yield of the transgenic tobacco with NtSSU gene were also significantly increased. The results revealed that overexpression of NtSSU gene could effectively redirect more photosynthesis carbon flux into starch biosynthesis pathway, which led to an increased biomass yield but did not generate negative effects on other agronomic traits. Therefore, NtSSU gene can be used as an excellent target gene in plant breeding to enrich starch accumulation in vegetative organs to develop new germplasm dedicated to fuel ethanol production.

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.

Socio-environmental risks associated with the green tobacco sickness in farmers: a case-control study / Riesgos socioambientales asociados a la enfermedad del tabaco verde en agricultores: un estudio caso-control / Riscos socioambientais associados à doença da folha verde do tabaco em agricultores: um estudo caso-controle

ABSTRACT Objective: to determine the presence of socio-environmental risk factors for the development of Green Tobacco Sickness in workers who grow Burley tobacco. Method: matched case-control study. The data collection took place in two moments: from December 2016 to January 2017 and December 2017, when the Burley tobacco was collected, through a household survey with interview application and urine collection for urinary cotinine. Results: the socio-environmental risk factors that remained associated with the disease were: bundling tobacco (p=0.047) and wearing socks (p=0.011); with protective effect were found sticking tobacco seeding of the day (p=0.006) and number of tobacco harvested per day (p=0.021). Conclusion: the steps in the Burley tobacco work process increase the exposure and risk of developing the disease. By identifying these factors, it is possible to address interdisciplinary control and prevention measures.

RESUMEN Objetivo: determinar la presencia de factores de riesgo socioambientales para el desarrollo de la Enfermedad del Tabaco Verde en trabajadores que cultivan el tabaco Burley. Método: estudio del tipo caso-control pareado. La recolección de datos ocurrió en dos momentos: de diciembre de 2016 a enero de 2017 y diciembre de 2017, período en que ocurrió la recolección del tabaco Burley, por medio de encuesta domiciliaria con aplicación de entrevista y recolección de orina para el examen de cotinina urinaria. Resultados: los factores de riesgo socioambientales que permanecieron asociados a la enfermedad fueron aferrar tabaco (p=0,047) y usar medias (p=0,011); con efecto protector fueron encontrados espetar los pies de tabaco del día (p=0,006) y el número de pies de tabaco cosechados por día (p=0,021). Conclusión: las etapas del proceso de trabajo con tabaco Burley aumentan la exposición y el riesgo de desarrollar la enfermedad. Al identificar estos factores, es posible dirigir, de forma interdisciplinaria, medidas de control y prevención.

RESUMO Objetivo: determinar a presença de fatores de riscos socioambientais para o desenvolvimento da Doença da Folha Verde do Tabaco em trabalhadores que cultivam o tabaco Burley. Método: estudo do tipo caso-controle pareado. A coleta de dados ocorreu em dois momentos: de dezembro de 2016 a janeiro de 2017 e dezembro de 2017, período em que ocorreu a colheita do tabaco Burley, por meio de inquérito domiciliar com aplicação de entrevista e coleta de urina para exame de cotinina urinária. Resultados: os fatores de riscos socioambientais que permaneceram associados à doença foram enfeixar tabaco (p=0,047) e usar meias (p=0,011); com efeito protetor foram encontrados espetar pés de tabaco do dia (p=0,006) e número de pés de tabaco colhido por dia (p=0,021). Conclusão: as etapas do processo de trabalho com tabaco Burley aumentam a exposição e o risco de desenvolver a doença. Ao identificar esses fatores, é possível direcionar, de forma interdisciplinar, medidas de controle e prevenção.

Antepenultimate residue at the C-terminus of NADPH oxidase RBOHD is critical for its function in the production of reactive oxygen species in Arabidopsis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Production of reactive oxygen species (ROS) is a conserved immune response primarily mediated by NADPH oxidases (NOXs), also known in plants as respiratory burst oxidase homologs (RBOHs). Most microbe-associated molecular patterns (MAMPs) trigger a very fast and transient ROS burst in plants. However, recently, we found that lipopolysaccharides (LPS), a typical bacterial MAMP, triggered a biphasic ROS burst. In this study, we isolated mutants defective in LPS-triggered biphasic ROS burst (delt) in Arabidopsis, and cloned the DELT1 gene that was shown to encode RBOHD. In the delt1-2 allele, the antepenultimate residue, glutamic acid (E919), at the C-terminus of RBOHD was mutated to lysine (K). E919 is a highly conserved residue in NADPH oxidases, and a mutation of the corresponding residue E568 in human NOX2 has been reported to be one of the causes of chronic granulomatous disease. Consistently, we found that residue E919 was indispensable for RBOHD function in the MAMP-induced ROS burst and stomatal closure. It has been suggested that the mutation of this residue in other NADPH oxidases impairs the protein's stability and complex assembly. However, we found that the E919K mutation did not affect RBOHD protein abundance or the ability of protein association, suggesting that the residue E919 in RBOHD might have a regulatory mechanism different from that of other NOXs. Taken together, our results confirm that the antepenultimate residue E is critical for NADPH oxidases and provide a new insight into the regulatory mechanisms of RBOHD.

Urinary cotinine in tobacco farmers in Southern Brazil

ABSTRACT OBJECTIVE To describe urinary cotinine levels in tobacco farmers. METHODS A cross-sectional study was conducted in 2,570 tobacco farmers. All participants that reported green tobacco sickness in the week prior to the interview plus a subsample of 492 pesticide applicators were included. We collected urinary samples and information about sociodemographic, behavioral, dietary, occupational characteristics, and pesticide poisoning during their lifetime. Stratification by sex and smoking was performed and the Wilcoxon and Kruskal-Wallis non-parametrical tests were used to analyze cotinine means. RESULTS This study included 582 individuals. There was no difference in urinary cotinine means between green tobacco sickness symptomatic and asymptomatic individuals. Among non-smokers, having picked tobacco in the previous week was associated with higher cotinine means in both genders. Cotinine levels were higher on the first day of symptoms and reduced exponentially with each day in female non-smokers. Male non-smokers had higher levels on the second day and a more gradual reduction. The cotinine level rose up to 15 cigarettes/day of consumption. CONCLUSIONS The urinary cotinine measures exposure to nicotine up to its saturation point; while green tobacco sickness, affected by tolerance, indicates nicotine poisoning. Strategies to reduce nicotine exposure in tobacco production are needed. Mechanization could be an alternative, as long as it overcame the challenge of irregular terrain and did not affect the quality of the leaf. More studies are needed to evaluate the chronic effect of nicotine exposure.

Fumar antecipa a menopausa?: evidências do estudo Pró-Saúde / Does smoking anticipate menopause? Pró-Saúde study evidence

A menopausa é definida pelo momento em que a menstruação cessa permanentemente como conseqüência da falência ovariana. Uma vez estabelecida, há um aumento no risco de doença coronariana, doença de Alzheimer, osteoporose e fraturas e sua antecipação está relacionada a maiores índices de mortalidade. Com o envelhecimento geral da população mundial, as mulheres passaram a viver de um terço a metade de suas vidas no período pósmenopáusico e, conseqüentemente, a pesquisa de condições associadas à menopausa ganham importância. A idade da menopausa e os fatores que a influenciam variam entre os diversos estudos e poucos estudos brasileiros abordam o tema. O fumo tem sido associado à antecipação da idade da menopausa. O objetivo deste estudo é analisar as diversas dimensões de associação entre o fumo e a idade da menopausa, levando em consideração possíveis relações de dose-resposta. Com base em dados do Estudo Pró-Saúde, foi realizado um estudo seccional utilizando-se para a análise o modelo de sobrevida paramétrico de riscos proporcionais com distribuição de Weibull. Os resultados apontam para uma redução em 32 por cento do risco de menopausa entre fumantes ativas, que a alcançam 2,5 anos mais tarde que nunca fumantes, ajustando-se para escolaridade e paridade. Entre fumantes ativas, no entanto, foram sugeridos aumentos de risco de 123 por cento e 192 por cento para fumantes de 10 a 20 cigarros por dia e de mais de 20 maços-ano respectivamente, quando comparadas às fumantes de menos de 10 cigarros por dia e menos de 10 maços-ano. Nesses grupos, a menopausa foi antecipada em 3,3 anos e 4,4 anos, respectivamente. Em relação ao tempo decorrido entre cessar o tabagismo e a idade menopausa, duração do fumo ou idade de início, não foram encontradas associações. Neste estudo, em possíveis efeitos de dose-resposta, o fumo apresenta-se como fator associado à antecipação da idade da menopausa em alguns aspectos, embora não em outros.

Purification and properties of antiviral proteins from the leaves of Bougainvillea xbuttiana.

A non-phytotoxic, resistance inducing, proteinaceous antiviral principle was purified by ammonium sulphate fractionation, ion exchange chromatography and gel filtration from the leaves of Bougainvillea xbuttiana. It imparted resistance against tobacco mosaic virus (TMV) and sunnhemp rosette virus (SRV) in their respective test hosts viz. Nicotiana glutinosa, N. tabacum var. Samsun NN, and Cyamopsis tetragonoloba, respectively. The purified principle eluted as a single peak upon gel filtration, but exhibited two polypeptides on SDS-PAGE with Mr 28,000 and 24,000. The two polypeptides were found to be highly basic, rich in lysine with pI around 10.0 and 10.5, respectively. Since this principle effected local lesion inhibition in both treated and untreated top leaves of test host, it might be acting in the initial stages of virus infection as a systemic inducer.

Assay of carcinogenicity of tobacco metabolites employing microsomal degranulation technique.

Carcinogenicity of salivarty extracts of different types of tobaccos smoked and chewed in India and Pan Parag were tested using microsomal degranulation technique. Results obtained on the basis of RNA/protein ratios (Indices to confirm the detachment of ribosomes from microsomes) showed that tobaccos used for cigarette, bidi, hukah and chewing tobacco with lime as well as Pan Parag were positively carcinogenic. Two fractions out of 7 isolated chromatographically from salivary extract of chewing tobacco plus lime were found to be carcinogenic. Elemental and spectral analyses indicated that the fractions are possibly an aromatic compound with an aliphatic side chain and N-(buty1 nitrosamine)-1-(3-pyridyl)-4-hydroxy-1-butanone.

Frecuencia del consumo de cigarrillo: Influencia de la publicidad La Paz-Septiembre-1991 / Cigarrette comsumption frecuency: Publishing influency in La Paz - Septiembre - 1991

Fumar cigarrillo produce enfermedades importantes que a su vez son causa de gran mortalidad en Latinoamerica. Bolivia esta considerada como buen mercado de las industrias tabacaleras. En la poblacion de la ciudad de La Paz, el consumo es importante en los niveles socioeconomicos altos. La influencia de la publicidad es muy importante para los fumadores. Elaboramos 312 cuestionarios personales con 25 preguntas en la poblacion de la ciudad de La Paz, de acuerdo a nivel socio-economico, edad y sexo: incluyendo estudiantes de medio, universitarios, empleados publicos, profesionales y trabajadores informales. Hay 13 por ciento de fumadores habituales y 26 por ciento ocasionales. La incidencia en estudiantes universitarios fue 24.6 por ciento . El porcentaje de inicio fue 54.1 por ciento en adolescentes. La encuesta concluye que la frecuencia de fumadores es elevada. El conocimiento del daño que produce es deficiente. La publicidad miente en afan de obtener mayor consumo. Recomendamos educacion sanitaria para la gente joven que debe ser constante, directa y adecuada

Some chemical and biological effects of cigarettes and Hubbell bubble smoking

The study was performed on 50 adult male cigarette and hubble smokers. The aim was to study and compare the cigarette and hubble bubble smoke on some chemical and biological functions of the smoke component. Plasma epinephrine, glucose, lactate/pyruvate ratio, growth hormone, free fatty acids, and cortisol were significantly higher in smokers than non smokers especially cigarette smokers. Plasma insulin concentration showed a significantly delayed response to the increased blood glucose. Plasma proteins, urea, pyruvate, creatinine and vitamin B12 were significantly reduced inn smokers than non smokers especially cigarette smokers. Both blood erythrocytes and leucocytes were increased significantly in smokers. Blood gases showed significantly low PaO2, significant blood level of Co fraction, while PaCO2 was unaffected. Urinary catecholamines were significantly higher in smokers especially in cigarette smokers