Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format

Vitamin A in the Vision of Insects

Acetone-methanol extracts of honeybees (Apis mellifera) were chromatographed from petroleum ether on columns of aluminum oxide and magnesium oxide:celite. Vitamin A1 was identified by the Carr-Price (antimony chloride) reaction. These experiments provide the first demonstration of vitamin A in the tissues of an insect. Like retinene, vitamin A is confined to the heads and is not found in either thoraces or abdomens. Dark-adapted bees have very little vitamin A. During light adaptation the vitamin A increases, but at the expense of retinene, which decreases. As much as 0.1 µg of vitamin A/gm of heads has been recovered from light-adapted bees. Two methods are described for demonstrating the enzymic reduction of retinene to vitamin A, using an extract of the heads of honeybees

Uso do SIG para estimar o potencial de distribuição geográfica de pragas quarentenárias em função de variáveis climáticas.

Praga quarentenária é uma praga de importância econômica potencial para uma área com risco de sua introdução, mas não presente? ou presente, mas não amplamente distribuído na área e estando oficialmente controlado. Um Sistema de Informações Geográficas pode ser utilizado como ferramenta na análise de risco de pragas a fim de avaliar o seu potencial de introdução e de dispersão em uma área após o seu estabelecimento. O objetivo deste estudo foi determinar o potencial de distribuição geográfica de duas espécies quarentenárias, a mosca oriental das frutas (Bactrocera dorsalis), e o mal seco (Deuterophoma tracheiphila ), no estado de São Paulo, com base em variáveis climáticas e a utilização da ferramenta de SIG. Como resultado deste estudo, ambas as pragas quarentenárias indicaram potencial de introdução e dispersão no estado de São Paulo

Pharmacogenomics and SSRIs Appropriateness in Older Community Dwelling African Americans

Background: Depressive and anxiety disorders are among the most common illnesses experienced by older adults (age \u3e 60). The selective serotonin reuptake inhibitors (SSRIs) are preferred class of antidepressants for these disorders due to their high efficacy and safety profiles among older adults. However, SSRIs are metabolized by highly polymorphic cytochrome P450 enzymes, specifically CYP2D6 and CYP2C19. This can lead to variable dose-response outcomes, especially among older African American population. Objective: Analyze the frequency of CYP2D6 and CYP2C19 polymorphisms in African American older adults who are taking SSRIs and identify potential inappropriate use of SSRIs in these older adults using the Clinical Pharmacogenetics Implementation Consortium (CPIC) guidelines. Methods: Participants (age \u3e 60) were enrolled into Translational Approaches to Personalized Health (TAPH) study. DNA samples were collected via Ora-gene saliva kits and the DNA was analyzed using the PGx Express Chip on the QuantStudio 12K Flex system. After quality control was performed, we focused on the genotypes of 12 participants who were prescribed SSRIs. Results: Only 2 participants had normal activity levels of both CYP2D6 and CYP2C19, while the rest had at least one variant allele that resulted in decreased or increased enzyme activity level. After matching the participants’ enzyme activity levels with the major metabolic pathway of their agent of SSRIs, 8 out of 12 participants are at risk of experiencing sub- or supra-therapeutic effects of SSRIs. 2 participants, especially, are at increased risk of serious adverse effect of citalopram-induced prolonged QT interval, which is more prevalent in older adults.https://scholarscompass.vcu.edu/gradposters/1148/thumbnail.jp

Assessment of in silico BAC-based simple sequence repeat (SSR) marker development for tomato (Solanum lycopersicum L.)

Tomato landraces are less sensitive to environmental stresses and grown mainly under rain fed conditions. They are still grown in small farms due to quality and special demand of consumers. These landraces are valuable sources of genetic traits, and plant breeders can use breeding programs for crop improvement. One of the primary needs of the crop improvement is the estimation of genetic diversity. Development of microsatellite simple sequence repeat (SSR) markers from map-referenced bacterial artificial chromosomes (BAC) clones is a very effective means of targeting markers to marker scarce positions in the genome. This study was aimed at developing a set of functional SSR markers via in silico analysis of publicly available tomato DNA sequences. As a result, 17 SSR markers were developed and tested on one tomato commercial cultivar and eight local landraces. 12 loci (27 alleles) were scored and showed 100% polymorphic patterns. The calculated polymorphism information content (PIC) values for the SSR markers developed ranged from 0.62 to 0.97 (mean 0.89). The SSR motifs CT(26) AT(27) and TTC(6) TTA(4) had the highest PIC value (0.97), while CAA(5)A(8) had the lowest PIC value (0.62). According to tomato expressed sequence tag (EST) analysis, some of these developed SSR markers, such as mono and di-nucleotide are related to some genes. The T(16) motif is related to hydroxyproline-rich glycoprotein, which is a family protein from Arabidopsis thaliana. On the other hand, the SSR with tri-nucleotide repeat motif AAC(4)A(11) was related to a putative homologous protein to A7Q2S4 from Vitis vinifera.Keywords: Tomato landraces, in silico simple sequence repeat (SSR) markers, DNA markers, genetic diversit

Avaliação das projeções de precipitação dos modelos climáticos globais do quarto relatório do IPCC para o Brasil.

Este trabalho tem como objetivo avaliar a similaridade das projeções de precipitação dos modelos climáticos globais para o Brasil e suas regiões. Quinze modelos globais do Quarto Relatório do IPCC foram avaliados, considerando suas projeções do período de 2071-2100, cenário A2 de emissão de gases de efeito estufa, e a precipitação observada no período de 1961-1990. Os resultados mostraram que os modelos globais climáticos se comportam de forma diferente para cada região brasileira, indicando tendência de aumento de precipitação em certas áreas geográficas e diminuição em outras. Nos meses de inverno, verifica-se uma tendência de diminuição das precipitações, enquanto no verão a tendência é de aumento das precipitações

Time course differential gene expression in response to porcine circovirus type 2 subclinical infection

This study was aimed at characterizing the potential differences in gene expression in piglets inoculated with Porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome. Seven-day-old caesarean-derived, colostrum-deprived piglets were distributed into two groups: control (n = 8) and pigs inoculated with 105.2 TCID50 of the Burgos PCV2 isolate (n = 16). One control and three inoculated pigs were necropsied on days 1, 2, 5, and 8 post-infection (p.i.). The remaining pigs (four of each group) were sequentially bled on days 0, 7, 14, 21, and 29 p.i. (necropsy). Total RNA from the mediastinal lymph node (MLN) and lysed whole blood (LWB) samples were hybridized to Affymetrix Porcine GeneChip®. Forty-three probes were differentially expressed (DE) in MLN samples (FDR < 0.1, fold change > 2) and were distributed into three clusters: globally down-regulated genes, and up-regulated genes at early (first week p.i.) and late (day 29 p.i.) stages of infection. In LWB samples, maximal differences were observed at day 7 p.i., with 54 probes DE between control and inoculated pigs. Main Gene Ontology biological processes assigned to up-regulated genes were related to the immune response. Six common genes were found in both types of samples, all of which belonged to the interferon signaling antiviral effector pathway. Down-regulated genes were mainly related to cell adhesion and migration in MLN, and cellular organization and biogenesis in LWB. Microarray results were validated by quantitative real-time PCR. This study provides, for the first time, the characterization of the early and late molecular events taking place in response to a subclinical PCV2 infection

Finite-key security against coherent attacks in quantum key distribution

The work by Christandl, K\"onig and Renner [Phys. Rev. Lett. 102, 020504 (2009)] provides in particular the possibility of studying unconditional security in the finite-key regime for all discrete-variable protocols. We spell out this bound from their general formalism. Then we apply it to the study of a recently proposed protocol [Laing et al., Phys. Rev. A 82, 012304 (2010)]. This protocol is meaningful when the alignment of Alice's and Bob's reference frames is not monitored and may vary with time. In this scenario, the notion of asymptotic key rate has hardly any operational meaning, because if one waits too long time, the average correlations are smeared out and no security can be inferred. Therefore, finite-key analysis is necessary to find the maximal achievable secret key rate and the corresponding optimal number of signals.Comment: 9 pages, 4 figure