Revealing the Chemical Composition of Birch Pollen Grains by Raman Spectroscopic Imaging

The investigation of the biochemical composition of pollen grains is of the utmost interest for several environmental aspects, such as their allergenic potential and their changes in growth conditions due to climatic factors. In order to fully understand the composition of pollen grains, not only is an in-depth analysis of their molecular components necessary but also spatial information of, e.g., the thickness of the outer shell, should be recorded. However, there is a lack of studies using molecular imaging methods for a spatially resolved biochemical composition on a single-grain level. In this study, Raman spectroscopy was implemented as an analytical tool to investigate birch pollen by imaging single pollen grains and analyzing their spectral profiles. The imaging modality allowed us to reveal the layered structure of pollen grains based on the biochemical information of the recorded Raman spectra. Seven different birch pollen species collected at two different locations in Germany were investigated and compared. Using chemometric algorithms such as hierarchical cluster analysis and multiple-curve resolution, several components of the grain wall, such as sporopollenin, as well as the inner core presenting high starch concentrations, were identified and quantified. Differences in the concentrations of, e.g., sporopollenin, lipids and proteins in the pollen species at the two different collection sites were found, and are discussed in connection with germination and other growth processes

EFEITO DA SAZONALIDADE SOBRE O RENDIMENTO DO ÓLEO ESSENCIAL DE Piper gaudichaudianum KUNTH

Piper gaudichaudianum Kunth (Piperaceae) is widely distributed throughout Brazil. In addition to its ecological importance, it is used in folk medicine and a great number of its biological activities have been already proved. Thus, the aim of this research was to investigate the effect of seasonality on the content of essential oil (EO) in leaves of Piper gaudichaudianum and ascertain the possible influence of the drying process of the plant material in the yield of this extractive. Leaves were randomly collected from individuals of a native population in Santa Maria, RS state, Brazil, along a year. The EO was extracted from fresh and dried leaves by hydrodistillation. When reproductive organs were observed, they were extracted in order to examine their influence on the yield of leaf EO. To determine the moisture content (MC%), 20 g of leaves and 10g of reproductive organs were dried separately in an oven at 50°C until constant weight. The yield of leaf EO (% w/ w) was calculated for the fresh material based on the dry weight considering the MC%, and for the dried material at room temperature. Only small seasonal variations in EO content was detected for the analyzed samples, with no statistical difference between them. The highest yield was observed in the spring for EO from the leaves (1.61 ± 0.09%), and from the reproductive organs (5.17%), while the lowest yields of leaf EO occurred in the summer (1.32 ± 0.14%), the only season which showed no presence of the reproductive organs. The absence of significant difference between the yields indicates no quantitative seasonal variability in production of EO in leaves of this population. However, yields did not differ statistically, indicating no seasonal variability of the EO content of the leaves of this population. Therefore, the collection of this plant organ can be performed at any time of the year in order to obtain this extractive. Since there was no decrease in the content of EO during the drying process at room temperature, this post-harvest processing can be adopted.Piper gaudichaudianum Kunth (Piperaceae) é uma planta com ampla distribuição no território brasileiro. Além de sua importância ecológica, é utilizada na medicina popular e muitas de suas atividades biológicas já foram comprovadas. O objetivo deste trabalho foi verificar o efeito da sazonalidade sobre o teor de óleo essencial (OE) em folhas de Piper gaudichaudianum e averiguar a possível influência do processo de secagem do material vegetal no rendimento deste extrativo. Foram coletadas aleatoriamente folhas de indivíduos de uma população nativa de Santa Maria - RS, Brasil, ao longo de um ano. O OE foi extraído de folhas frescas e secas à temperatura ambiente pelo método de hidrodestilação. Quando verificada a presença de órgãos reprodutivos, os mesmos foram extraídos, a fim de examinar a sua influência no rendimento do OE de folhas. Para determinação do teor de umidade (TU%), 20 g de folhas e 10 g de órgãos reprodutivos foram secos separadamente em estufa a 50°C até peso constante. O rendimento de OE de folhas (% m/m) foi calculado para o material fresco com base na massa seca, considerando o TU%, bem como para o material seco à temperatura ambiente. Apenas pequenas variações sazonais nos teores de OE foram detectadas para as amostras analisadas, não havendo diferença estatística entre eles. O maior rendimento foi observado na primavera, tanto para o OE de folhas (1,61±0,09%), quanto de órgãos reprodutivos (5,17%), enquanto o menor teor de OE em folhas ocorreu no verão (1,32±0,14%), a única estação em que não houve a presença de órgãos reprodutivos. A ausência de diferença significativa entre os rendimentos indica que não houve variabilidade sazonal quantitativa na produção de OE em folhas desta população. Portanto, a coleta deste órgão vegetal pode ser realizada em qualquer época do ano visando à obtenção deste extrativo. Uma vez que não foi verificada diminuição do teor de OE durante o processo de secagem à temperatura ambiente, esse processamento pós-colheita poderá ser adotado

Genetic inactivation of the Fanconi anemia gene FANCC identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents

Background: Inactivation of the Fanconi anemia (FA) pathway through defects in one of 13 FA genes occurs at low frequency in various solid cancer entities among the general population. As FA pathway inactivation confers a distinct hypersensitivity towards DNA interstrand-crosslinking (ICL)-agents, FA defects represent rational targets for individualized therapeutic strategies. Except for pancreatic cancer, however, the prevalence of FA defects in gastrointestinal (GI) tumors has not yet been systematically explored. Results: A panel of GI cancer cell lines was screened for FA pathway inactivation applying FANCD2 monoubiquitination and FANCD2/RAD51 nuclear focus formation and a newly identified FA pathway-deficient cell line was functionally characterized. The hepatocellular carcinoma (HCC) line HuH-7 was defective in FANCD2 monoubiquitination and FANCD2 nuclear focus formation but proficient in RAD51 focus formation. Gene complementation studies revealed that this proximal FA pathway inactivation was attributable to defective FANCC function in HuH-7 cells. Accordingly, a homozygous inactivating FANCC nonsense mutation (c.553C > T, p.R185X) was identified in HuH-7, resulting in partial transcriptional skipping of exon 6 and leading to the classic cellular FA hypersensitivity phenotype; HuH-7 cells exhibited a strongly reduced proliferation rate and a pronounced G2 cell cycle arrest at distinctly lower concentrations of ICL-agents than a panel of non-isogenic, FA pathway-proficient HCC cell lines. Upon retroviral transduction of HuH-7 cells with FANCC cDNA, FA pathway functions were restored and ICL-hypersensitivity abrogated. Analyses of 18 surgical HCC specimens yielded no further examples for genetic or epigenetic inactivation of FANCC, FANCF, or FANCG in HCC, suggesting a low prevalence of proximal FA pathway inactivation in this tumor type. Conclusions: As the majority of HCC are chemoresistant, assessment of FA pathway function in HCC could identify small subpopulations of patients expected to predictably benefit from individualized treatment protocols using ICL-agents

Variabilidad de la composición química del aceite esencial de Nectandra megapotamica (Spreng.) Mez (Lauraceae)

Este trabajo describe la variabilidad química del aceite esencial (AE) de N. megapotamica, utilizando análisis multivariado. Se colectaron hojas de tres individuos en una población en Santa Maria, Brasil, en cada estación durante un año y se clasificaron en hojas jóvenes y viejas. Los AE se obtuvieron por hidrodestilación y la composición química fue determinada por cromatografía de gases acoplada a espectrometría de masas. Se realizó un análisis de agrupamiento jerárquico para formación de grupos químicos (GQ) y análisis de componentes principales (ACP) para verificar cuáles constituyentes influenciaron la formación de grupos. Inicialmente se observaron dos agrupamientos en el dendrograma debido a concentraciones de α-pineno y biciclogermacreno, constituyentes mayoritarios. En el segundo corte se discriminaron cinco GQ, siendo este también el número de componentes principales con valores propios superiores a 1. Los isómeros de asarona se observaron solamente en el AE de dos individuos (GQ5), constituyendo los principales elementos de diferenciación

Spitzer 24 micron detections of starburst galaxies in Abell 851

Spitzer-MIPS 24 micron and ground-based observations of the rich galaxy cluster Abell 851 at z=0.41 are used to derive star formation rates from the mid-IR 24 micron and from [O II] 3727 emission. Many cluster galaxies have SFR(24 um)/SFR([O II]) >> 1, indicative of star formation highly obscured by dust. We focus on the substantial minority of A851 cluster members where strong Balmer absorption points to a starburst on a 10^{8-9} year timescale. As is typical, A851 galaxies with strong Balmer absorption occur in two types: with optical emission (starforming), and without (post-starburst). Our principal result is the former, so-called e(a) galaxies, are mostly detected (9 out of 12) at 24 um -- for these we find typically SFR(24 um)/SFR([O II]) ~ 4. Strong Balmer absorption and high values of SFR(24 um)/SFR([O II]) both indicate moderately active starbursts and support the picture that e(a) galaxies are the active starbursts that feed the post-starburst population. While 24 um detections are frequent with Balmer-strong objects (even 6 out of 18 of the supposedly "post-starburst'' galaxies are detected) only 2 out of 7 of the continuously starforming `e(c)' galaxies (with weak Balmer absorption) are detected -- for them, SFR(24 um)/SFR([O II]) ~ 1. Their optical spectra resemble present-epoch spirals that dominate today's universe; we strengthen this association by that SFR(24 um)/SFR([O II]) ~ 1 is the norm today. That is, not just the amount of star formation, but its mode, has evolved strongly from z ~ 0.4 to the present. By fitting spectrophotometric models we measure the strength and duration of the bursts to quantify the evolutionary sequence from active- to post-starburst, and to harden the evidence that moderately active starbursts are the defining feature of starforming cluster galaxies at z ~ 0.4.Comment: Accepted to The Astrophysical Journal. Abstract abridge

CTNSmRNA as a potential treatment for nephropathic cystinosis

Messenger RNA (mRNA) therapies are emerging in different disease areas, but have not yet reached the kidney field. Our aim was to study the feasibility to correct the genetic defect in nephropathic cystinosis using synthetic mRNA. Cystinosis is a prototype disorder of proximal tubular dysfunction caused by mutations in the CTNS gene, encoding the lysosomal cystine-H+ symporter cystinosin, and leading to cystine accumulation in all cells of the body. The kidneys are the first and most severely affected organs, presenting glomerular and proximal tubular dysfunction. Cysteamine is the current therapeutic standard that reduces cellular cystine levels, but has many side effects and does not restore kidney function. Here, we show that synthetic mRNA is safe and effective to reintroduce functional cystinosin using lipofection in CTNS-/- kidney cells and following direct injection in ctns-/- zebrafish larvae. CTNS mRNA therapy results in prompt lysosomal expression of the functional protein and decreases cellular cystine accumulation for up to 14 days. In the ctns-/- zebrafish, CTNS mRNA therapy improves proximal tubular reabsorption, reduces proteinuria, and restores brush border expression of the multi-ligand receptor megalin. We propose that mRNA-based therapy, if sufficient kidney targeting can be achieved, may be a new approach to treat cystinosis