Mapping of a Leishmania major gene/locus that confers pentamidine resistance by deletion and insertion of transposable element

Pentamidine (PEN) is an alternative compound to treat antimony-resistant leishmaniasis patients, which cellular target remains unclear. One approach to the identification of prospective targets is to identify genes able to mediate PEN resistance following overexpression. Starting from a genomic library of transfected parasites bearing a multicopy episomal cosmid vector containing wild-type Leishmania major DNA, we isolated one locus capable to render PEN resistance to wild type cells after DNA transfection. In order to map this Leishmania locus, cosmid insert was deleted by two successive sets of partial digestion with restriction enzymes, followed by transfection into wild type cells, overexpression, induction and functional tests in the presence of PEN. To determine the Leishmania gene related to PEN resistance, nucleotide sequencing experiments were done through insertion of the transposon Mariner element of Drosophila melanogaster (mosK) into the deleted insert to work as primer island. Using general molecular techniques, we described here this method that permits a quickly identification of a functional gene facilitating nucleotide sequence experiments from large DNA fragments. Followed experiments revealed the presence of a P-Glycoprotein gene in this locus which role in Leishmania metabolism has now been analyzed.A Pentamidina (PEN) é um composto alternativo para o tratamento de pacientes com leishmaniose que apresentam resistência ao antimônio, cujo alvo celular continua incerto. Uma abordagem para se identificar prováveis alvos seria a identificação e super-expressão de genes capazes de mediar resistência a PEN. A partir de uma genoteca construída com o DNA de Leishmania major em um vetor - cosmídio que se desenvolve tanto em bactérias como nas células do parasita, isolamos um locus que após transfecção é capaz de produzir resistência a PEN às células do parasita. Almejando o mapeamento desse locus de leishmania, o inserto clonado nesse cosmídio foi deletado através de duas digestões parciais sucessivas com enzimas de restrição, seguida de transfecção em células selvagens, super-expressão gênica, indução e testes funcionais na presença de PEN. Para determinar o gene de Leishmania relacionado com a resistência a PEN, o seqüenciamento de nucleotídeos foi executado após inserção de elementos transposicionais de Drosophila melanogaster no interior do inserto deletado para atuar como 'ilhas de iniciadores'. Descrevemos aqui o mapeamento desse locus, após a inserção transposicional, que além de facilitar o seqüenciamento de nucleotídeos de grandes fragmentos de DNA, permite uma rápida identificação do gene relacionado com esse fenótipo. Experimentos posteriores revelaram neste locus a presença do gene de uma Glicoproteína-P de membrana, cujo papel no metabolismo na Leishmania está sendo analisado

Fiber Bragg Grating (FBG) Sensors in a High-Scattering Optical Fiber Doped with MgO Nanoparticles for Polarization-Dependent Temperature Sensing

International audienceFeatured Application: Inscription and interrogation of fiber Bragg gratings into MgO nanoparticle-doped fiber for optical fiber distributed and multiplexed sensing. Abstract: The characterization of Fiber Bragg Grating (FBG) sensors on a high-scattering fiber, having the core doped with MgO nanoparticles for polarization-dependent temperature sensing is reported. The fiber has a scattering level 37.2 dB higher than a single-mode fiber. FBGs have been inscribed by mean of a near-infrared femtosecond laser and a phase mask, with Bragg wavelength around 1552 nm. The characterization shows a thermal sensitivity of 11.45 pm/ • C. A polarization-selective thermal behavior has been obtained, with sensitivity of 11.53 pm/ • C for the perpendicular polarization (S) and 11.08 pm/ • C for the parallel polarization (P), thus having 4.0% different sensitivity between the two polarizations. The results show the inscription of high-reflectivity FBGs onto a fiber core doped with nanoparticles, with the possibility of having reflectors into a fiber with tailored Rayleigh scattering properties

Presence of Clostridium in corn silage cut at different stages of maturity

O milho (Zea mays L.) foi ensilado em sacos de polietileno em cinco diferentes estádios de maturidade e de umidade com o objetivo de determinar o número de esporos de bactérias do gênero Clostridium presentes. Paralelamente, foram determinados os parâmetros químicos de avaliação das silagens: teor de proteínas, matéria seca, umidade, pH e NH3/N. Pelos resultados obtidos concluiu-se que o milho apresentou teor ótimo de matéria seca para ensilagem (35%) entre 15 e 16 semanas após o plantio, no estádio de grãos farináceos. A elevada acidez da massa ensilada (pH = 3,96 - 4,10) foi responsável pela inibição da germinação dos esporos de Clostridium sp. presentes. O número de esporos viáveis na massa ensilada diminuiu à medida que aumentou o teor de matéria seca, com coeficiente de correlação (r = - 0,7949**) estatisticamente significativo.Com (Zea mays L.) was ensiled in plastic bags at five maturity stages and different moisture contents with the objective of determining the number of Clostridium spores present. Chemical analyses were made in the silage material to determine the protein content, dry matter, moisture, pH and NH3/N. According to the results obtained, it was concluded that the corn presented an optimum level of dry matter (35%) for ensilage between 15 and 16 weeks after planting, in the stage of farinaceous grains. The high acidity of the ensiled mass (pH 3,96 - 4,10) was responsible for the inhibition of the germination of Clostridium sp. spores. The number of viable spores in the ensiled mass decreased with the increase of the dry matter content with the correlation coefficient (r = -0,7949**) statistically significant

Largely tunable dispersion chirped polymer FBG

[EN] We demonstrate a largely tunable dispersion fiber Bragg grating (FBG) inscribed in a microstructured polymer optical fiber (mPOF). The bandwidth of the chirped FBG (CFBG) was achieved from 0.11 to 4.86 nm, which corresponds to a tunable dispersion range from 513.6 to 11.15 ps/nm. Furthermore, thermal sensitivity is used to compensate for the wavelength shift due to the applied strain. These results demonstrate that a CFBG in a POF is a promising technology for future optical systems. (C) 2018 Optical Society of AmericaFundacao para a Ciencia e a Tecnologia (FCT) (SFRH/BPD/109458/2015, UID/EEA/50008/2013); Research Excellence Award Programme GVA (PROMETEO 2017/103); Oak Ridge Associated Universities (ORAU) (LIFESTART (2017-2019)); Natural Science Foundation of Heilongjiang Province (F2018026).Min, R.; Korganbayev, S.; Molardi, C.; Broadway, C.; Hu, X.; Caucheteur, C.; Bang, O.... (2018). Largely tunable dispersion chirped polymer FBG. Optics Letters. 43(20):5106-5109. https://doi.org/10.1364/OL.43.005106S510651094320Reyes, P. I., Litchinitser, N., Sumetsky, M., & Westbrook, P. S. (2005). 160-Gb/s tunable dispersion slope compensator using a chirped fiber Bragg grating and a quadratic heater. IEEE Photonics Technology Letters, 17(4), 831-833. doi:10.1109/lpt.2005.843690Tang, Z., Pan, S., Zhu, D., Guo, R., Zhao, Y., Pan, M., … Yao, J. (2012). Tunable Optoelectronic Oscillator Based on a Polarization Modulator and a Chirped FBG. IEEE Photonics Technology Letters, 24(17), 1487-1489. doi:10.1109/lpt.2012.2205235He, X., Liu, Z., & Wang, D. N. (2012). Wavelength-tunable, passively mode-locked fiber laser based on graphene and chirped fiber Bragg grating. Optics Letters, 37(12), 2394. doi:10.1364/ol.37.002394Zhou, W., Dong, X., Ni, K., Chan, C. C., & Shum, P. (2010). Temperature-insensitive accelerometer based on a strain-chirped FBG. Sensors and Actuators A: Physical, 157(1), 15-18. doi:10.1016/j.sna.2009.11.003Tosi, D., Macchi, E. G., Gallati, M., Braschi, G., Cigada, A., Rossi, S., … Lewis, E. (2014). Fiber-optic chirped FBG for distributed thermal monitoring of ex-vivo radiofrequency ablation of liver. Biomedical Optics Express, 5(6), 1799. doi:10.1364/boe.5.001799Ortega, B., Cruz, J. L., Capmany, J., Andres, M. V., & Pastor, D. (2000). Variable delay line for phased-array antenna based on a chirped fiber grating. IEEE Transactions on Microwave Theory and Techniques, 48(8), 1352-1360. doi:10.1109/22.859480Wang, C., & Yao, J. (2008). Photonic Generation of Chirped Millimeter-Wave Pulses Based on Nonlinear Frequency-to-Time Mapping in a Nonlinearly Chirped Fiber Bragg Grating. IEEE Transactions on Microwave Theory and Techniques, 56(2), 542-553. doi:10.1109/tmtt.2007.914639Capmany, J., Ortega, B., & Pastor, D. (2006). A tutorial on microwave photonic filters. Journal of Lightwave Technology, 24(1), 201-229. doi:10.1109/jlt.2005.860478Lauzon, J., Thibault, S., Martin, J., & Ouellette, F. (1994). Implementation and characterization of fiber Bragg gratings linearly chirped by a temperature gradient. Optics Letters, 19(23), 2027. doi:10.1364/ol.19.002027Hill, P. C., & Eggleton, B. J. (1994). Strain gradient chirp of fibre Bragg gratings. Electronics Letters, 30(14), 1172-1174. doi:10.1049/el:19940772Kashyap, R., McKee, P. F., Williams, D. L., & Campbell, R. J. (1994). Novel method of producing all fibre photoinduced chirped gratings. Electronics Letters, 30(12), 996-998. doi:10.1049/el:19940669Candiani, A., Margulis, W., Sterner, C., Konstantaki, M., & Pissadakis, S. (2011). Phase-shifted Bragg microstructured optical fiber gratings utilizing infiltrated ferrofluids. Optics Letters, 36(13), 2548. doi:10.1364/ol.36.002548Marques, C. A. F., Webb, D. J., & Andre, P. (2017). Polymer optical fiber sensors in human life safety. Optical Fiber Technology, 36, 144-154. doi:10.1016/j.yofte.2017.03.010Hejie Yang, Lee, S. C. J., Tangdiongga, E., Okonkwo, C., van den Boom, H., Breyer, F., … Koonen, A. (2010). 47.4 Gb/s Transmission Over 100 m Graded-Index Plastic Optical Fiber Based on Rate-Adaptive Discrete Multitone Modulation. Journal of Lightwave Technology, 28(4), 352-359. doi:10.1109/jlt.2009.2034393Xiong, Z., Peng, G. D., Wu, B., & Chu, P. L. (1999). Highly tunable Bragg gratings in single-mode polymer optical fibers. IEEE Photonics Technology Letters, 11(3), 352-354. doi:10.1109/68.748232Pereira, L. M., Pospori, A., Antunes, P., Domingues, M. F., Marques, S., Bang, O., … Marques, C. A. F. (2017). Phase-Shifted Bragg Grating Inscription in PMMA Microstructured POF Using 248-nm UV Radiation. Journal of Lightwave Technology, 35(23), 5176-5184. doi:10.1109/jlt.2017.2771436Hu, X., Pun, C.-F. J., Tam, H.-Y., Mégret, P., & Caucheteur, C. (2014). Tilted Bragg gratings in step-index polymer optical fiber. Optics Letters, 39(24), 6835. doi:10.1364/ol.39.006835Lacraz, A., Polis, M., Theodosiou, A., Koutsides, C., & Kalli, K. (2015). Femtosecond Laser Inscribed Bragg Gratings in Low Loss CYTOP Polymer Optical Fiber. IEEE Photonics Technology Letters, 27(7), 693-696. doi:10.1109/lpt.2014.2386692Hongbo Liu, Huiyong Liu, Gangding Peng, & Whitbread, T. W. (2005). Tunable dispersion using linearly chirped polymer optical fiber Bragg gratings with fixed center wavelength. IEEE Photonics Technology Letters, 17(2), 411-413. doi:10.1109/lpt.2004.839378Marques, C. A. F., Antunes, P., Mergo, P., Webb, D. J., & Andre, P. (2017). Chirped Bragg Gratings in PMMA Step-Index Polymer Optical Fiber. IEEE Photonics Technology Letters, 29(6), 500-503. doi:10.1109/lpt.2017.2662219Min, R., Ortega, B., & Marques, C. (2018). Fabrication of tunable chirped mPOF Bragg gratings using a uniform phase mask. Optics Express, 26(4), 4411. doi:10.1364/oe.26.004411Cruz, J. L., Dong, L., Barcelos, S., & Reekie, L. (1996). Fiber Bragg gratings with various chirp profiles made in etched tapers. Applied Optics, 35(34), 6781. doi:10.1364/ao.35.006781Hu, X., Woyessa, G., Kinet, D., Janting, J., Nielsen, K., Bang, O., & Caucheteur, C. (2017). BDK-doped core microstructured PMMA optical fiber for effective Bragg grating photo-inscription. Optics Letters, 42(11), 2209. doi:10.1364/ol.42.002209Saez-Rodriguez, D., Min, R., Ortega, B., Nielsen, K., & Webb, D. J. (2016). Passive and Portable Polymer Optical Fiber Cleaver. IEEE Photonics Technology Letters, 28(24), 2834-2837. doi:10.1109/lpt.2016.2623419Hu, X., Pun, C.-F. J., Tam, H.-Y., Mégret, P., & Caucheteur, C. (2014). Highly reflective Bragg gratings in slightly etched step-index polymer optical fiber. Optics Express, 22(15), 18807. doi:10.1364/oe.22.018807Broadway, C., Gallego, D., Pospori, A., Zubel, M., Webb, D. J., Sugden, K., … Lamela, H. (2016). Microstructured polymer optical fibre sensors for opto-acoustic endoscopy. Micro-Structured and Specialty Optical Fibres IV. doi:10.1117/12.2227588Dong, L., Cruz, J. L., Reekie, L., & Tucknott, J. A. (1995). Fabrication of chirped fibre gratings using etched tapers. Electronics Letters, 31(11), 908-909. doi:10.1049/el:19950588Yuan, W., Stefani, A., Bache, M., Jacobsen, T., Rose, B., Herholdt-Rasmussen, N., … Bang, O. (2011). Improved thermal and strain performance of annealed polymer optical fiber Bragg gratings. Optics Communications, 284(1), 176-182. doi:10.1016/j.optcom.2010.08.069Erdogan, T. (1997). Fiber grating spectra. Journal of Lightwave Technology, 15(8), 1277-1294. doi:10.1109/50.618322Bai-Ou Guan, Hwa-Yaw Tam, Xiao-Ming Tao, & Xiao-Yi Dong. (2000). Simultaneous strain and temperature measurement using a superstructure fiber Bragg grating. IEEE Photonics Technology Letters, 12(6), 675-677. doi:10.1109/68.84908

High accuracy monitoring of honey bee colony development by a quantitative method

Honey bees are key insect pollinators, providing important economic and ecological value for human beings and ecosystems. This has triggered the development of several monitoring methods for assessing the temporal development of colony size, food storage, brood and pathogens. Nonetheless, most of these methods are based on visual assessments that are observer-dependent and prone to bias. Furthermore, the impact on colony development (invasiveness), as well as accuracy, were rarely considered when implementing new methods. In this study, we present and test a novel accurate and observer-independent method for honey bee colony assessment, capable of being fully standardized. Honey bee colony size is quantified by assessing the weight of adult bees, while brood and provision are assessed by taking photos and conducting image analysis of the combs with the image analysis software DeepbeeVR . The invasiveness and accuracy of the method were investigated using field data from two experimental apiaries in Portugal, comparing results from test and control colonies. At the end of each field experiment, most of the tested colonies had the same colony size, brood levels and honey production as the control colonies. Nonetheless, continuous weight data indicated some disturbance in tested colonies in the first year of monitoring. The overall accuracy of the image analysis software was improved by training, indicating that it is possible to adapt the software to local conditions. We conclude that the use of this fully quantitative method offers a more accurate alternative to classic visual colony assessments, with negligible impact on colony development.This work was supported by European Food Safety Authority under grant OC/EFSA/SCER/2017/02; FCT provided financial support by national funds (FCT/MCTES) to CFE (UIDB/04004/2020) and CIMO (UIDB/00690/2020); NC was financed by FCT under PhD grant SFRH/BD/133352/2017; YLD by DCE (Danish Centre for Environment and Energy) under grant 21628-82105.info:eu-repo/semantics/publishedVersio

Avaliação de duas variedades de cana-de-açúcar submetidas a diferentes tempos de armazenamento

A study was carried out to investigate the effects of four different storage times on technology and bromatologic characteristics in two sugar cane varieties (CO 413 and RB 72 454). The varieties were cut and stored during zero, 1.5, 3.0 and 4.5 days. It was used a completely randomized design according to a 2 x 4 factorial scheme. The RB 72 454 variety showed better dry matter level than the CO 413 variety. The storage time increased (P < 0.01) the reducters sugar levels and decreased (P < 0.01) the crude protein levels. When they were stored during 4.5 days, a significative (P < 0.01) reduction of 14.66 and 4.04% in the levels of crude protein and gross energy, respectively, was observed.Foram estudadas duas variedades de cana-de-açúcar (CO 413 e RB 72 454), com o objetivo de verificar o efeito do armazenamento pós-corte sobre suas características tecnológicas e bromatológicas. As variedades foram cortadas e armazenadas durante zero, 1,5, 3,0 e 4,5 dias. Utilizou-se o delineamento inteiramente casualizado em esquema fatorial 2 x 4 . A variedade RB 72 454 apresentou melhor maturação que a variedade CO 413. O armazenamento proporcionou aumento no teor de açúcares redutores (P < 0,01) e diminuição no teor protéico (P < 0,01) das variedades de cana em estudo. A variedade RB 72 454 apresentou maior teor de matéria seca (P < 0,01). Ambas as variedades, quando armazenadas durante 4,5 dias, apresentaram queda de 14,66 e 4,04% nos teores de proteína bruta e energia bruta, respectivamente

Efeito dos movimentos sacádicos horizontal e vertical dos olhos sobre o controle postural de adultos jovens e idosos em diferentes bases de apoio

O objetivo deste estudo foi investigar o efeito dos movimentos sacádicos, horizontal e vertical, dos olhos no desempenho do controle postural de adultos jovens e idosos durante a manutenção da postura em pé em diferentes bases de apoio. Quinze adultos jovens e quinze idosos permaneceram parados em pé, sobre uma plataforma de força, nas bases de apoio bipodal e semi tandem, durante a execução de duas tarefasvisuais, fixação de um alvo e movimentos sacádicos dos olhos nas direções horizontal e vertical. O Centro de Pressão (CP) foi medido nas direções de movimento anteroposterior (AP) e médio-lateral (ML). Os resultados mostraram que a frequência AP e a área de deslocamento do CP foram menores durante os movimentos sacádicos do que durante a fixação do alvo somente para os adultos jovens. Os resultados também revelaramque os idosos tiveram maior velocidade e frequência ML de deslocamento do CP do que os adultos jovens durante a base de apoio semi tandem. Não houve efeito de direção dos movimentos sacádicos (horizontal e vertical) para adultos jovens e idosos. Os resultados sugerem que os idosos não foram capazes de realizar os ajustes posturais necessários às tarefas visuais envolvendo os movimentos sacádicos, contribuindo parauma maior instabilidade postural em comparação aos adultos jovens

Effects of zeranol and A, D and E vitamins. I. On the performance of finishing crossbred steers.

Foram utilizados 64 animais mestiços Guzerá x Holandês e Guzerá x Charolês, com idade média de 21,4 ± 0,6 meses e peso vivo médio de 361,2 ± 11,6 kg, para estudar os efeitos do implante do zeranol e da ministração de vitaminas A, D e E sobre o desempenho de bovinos em confinamento durante a fase de terminação. O delineamento estatístico foi em blocos casualizados, em esquema fatorial 23, com dois tipos de mestiçagem, dois níveis de vitaminas (0 e 5,0 ml/cabeça) e dois níveis de anabolizante (0 e 36,0 mg de zeranol/cabeça), com oito repetições. Os animais foram alimentados com silagem de milho, à vontade, e 4,0 kg de grão de sorgo moídos, ambos oferecidos diariamente, em duas refeições, além de 1,0 kg da mistura melaço e uréia (9:1) e 100 gramas de sal mineralizado. Os melhores ganhos em peso (P < 0,01) foram obtidos com os mestiços Guzerá x Holandês (1,29 kg/cabeça/dia), em comparação com os mestiços Guzerá x Charolês (1,16 kg/cabeça/dia). Os animais que receberam o anabolizante apresentaram ganhos da ordem de 1,27 kg/cabeça/dia, superiores (P < 0,05) àqueles que não receberam (1,18 kg/cabeça/dia). A aplicação das vitaminas não melhorou o ganho em peso dos animais, cujos valores foram 1,22 e 1,22 kg/dia para os animais com e sem vitaminas A, D e E, respectivamente.Sixty-four crossbred animals Guzera x Holstein and Guzera x charolais, with average age of 21.4 ± 0,6 months and liveweight of 361.2 ± 11.6 kg were used in order to study the effocts of zeranol implant and A, D, and E vitamins minisiration, on the performance of finishing crassbred steers confined during the finishing period. The statistical design was in randamized blocks, in a factorial arrangement 2³, with two types of crossbreeding, two levels of vitamins (0 and 5.0 ml/animal) and two levels of anabolic agent (0 and 36.0 mg of zeranol/animal), with eight replications. The animal were fed with corn silage 'ad libitum', 4.0 kg of ground cron grain, both offered daily, in two meals, besides 1.0 kg of molasses and urea mixture (9:1), and 100 g of mineralized salt. The best liveweight gains (P < 0.01) were obtained with thee Guzera x Holstein crossbreds (1.29 kg/animal/day), compared to Guzera x Charolais crossbreds (1.16 kg/animal/day). The animals that received the anabolic agent showed gains of 1.27 kg/animal/day, higher than those that did not ceive it (1.18 kg/animal/day). The application of vitamins did not affect me weight gain of the animals, whose values were 1.22 and 1.22 kg/day to the animals with and without vitamins A, D, and E, respectively

Effects of zeranol and A, D and E vitamins on performance of Guzerá and crossbred with European cattle

O experimento foi realizado no confinamento da Faculdade de Ciências Agrárias e Veterinárias, Campus de Jaboticabal, da UNESP, com o objetivo de estudar o efeito do implante com zeranol e da aplicação das vitaminas A, D e E sobre o desempenho de bovinos da raça Guzerá e mestiços Europeu x Guzerá e os possíveis efeitos sobre o consumo de matéria seca, proteína bruta, energia bruta e conversão alimentar. Foi utilizado o delineamento em blocos ao acaso em esquema fatorial 2 x 3 com dois tipos de animais: (Guzerá e mestiços Europeu x Guzerá) e três tratamentos (0, 5,0 ml das vitaminas A, D e E e 36,0 mg de zeranol/animal, com cinco repetições por tratamento. O implante com zeranol melhorou o ganho de peso em 10,9%. A aplicação das vitaminas A, D e E não influenciou (P > 0,05) o ganho de peso. Os animais mestiços consumiram mais alimento por unidade de ganho de peso (P 0.05). The crossbred animals consumed significantly (P < 0.05) more feed per weight gain unit than Guzerá cattle