Validation of Tuba1a as Appropriate Internal Control for Normalization of Gene Expression Analysis during Mouse Lung Development

The expression ratio between the analysed gene and an internal control gene is the most widely used normalization method for quantitative RT-PCR (qRT-PCR) expression analysis. The ideal reference gene for a specific experiment is the one whose expression is not affected by the different experimental conditions tested. In this study, we validate the applicability of five commonly used reference genes during different stages of mouse lung development. The stability of expression of five different reference genes (Tuba1a, Actb Gapdh, Rn18S and Hist4h4) was calculated within five experimental groups using the statistical algorithm of geNorm software. Overall, Tuba1a showed the least variability in expression among the different stages of lung development, while Hist4h4 and Rn18S showed the maximum variability in their expression. Expression analysis of two lung specific markers, surfactant protein C (SftpC) and Clara cell-specific 10 kDA protein (Scgb1a1), normalized to each of the five reference genes tested here, confirmed our results and showed that incorrect reference gene choice can lead to artefacts. Moreover, a combination of two internal controls for normalization of expression analysis during lung development will increase the accuracy and reliability of results.     

Löschen im Internet – rechtlich gefordert und technisch möglich!

Die im Art. 17 der Datenschutz-Grundverordnung (DSGVO) enthaltene Regelungen für das Löschen, insbesondere die Pflicht, Dritte über die Löschanfragen zu informieren, sind mit einem hohen Aufwand für die Verantwortlichen verbunden, sodaßs sie ohne eine technische Unterstützung in der Praxis nicht umgesetzt werden können. Die zurzeit jedoch fehlenden technischen Lösungen führen zu einer Lücke zwischen den Rechtsvorschriften und den derzeitigen technischen Möglichkeiten, während die DSGVO ab Mai 2018 anzuwenden ist. Um diese Lücke zu schließen, wird im Folgenden zunächst analysiert, welche Möglichkeiten im Allgemeinen bestehen, das Löschen im Internet gemäß Art. 17 technisch umzusetzen. Anschließend wird eine Lösung für die Verteilung der Löschanfragen an die Verantwortlichen vorgestellt, das auf der Kombination der Technik mit dem Gesetz beruht und sowohl den Betroffenen als auch den Verantwortlichen ein Werkzeug zur Unterstützung des Art. 17 zur Verfügung stellt.     

Effect of height on the corrosion of steel and galvanized steel in nontropical coastal marine environments

In this study, the behavior of carbon steel and galvanized steel in nontropical coastal marine environments was evaluated. Evaluation was carried out with specimens with dimensions of 10 cm × 10 cm × 0.3 cm. These specimens were exposed to four testing stations (Iquique, Mejillones, Los Vilos, and San Vicente), where racks were installed both at ground level (ground), as well as in the upper zone of electrical transmission towers (tower). In each station, 24 specimens of A36 carbon steel and galvanized steel were placed (12 each). The corrosivity of the environment was measured using the ISO 9223, 9225, and 9226 standards. The specimens were evaluated on-site, monthly, through visual inspection and photographic record. Once withdrawn, the corrosion rate was determined and the corrosion products were analyzed through Raman and Fourier-transform infrared. The results show that, in all cases, the corrosion rate is greater in the tower than on the ground. However, even though the Los Vilos station is located farther from the sea (3,500 vs. ≈500 m), the corrosion rate of steel in the tower is the highest. This is caused by the generation of HCl from the transformation of lepidocrocite into goethite, in the presence of low chloride content, which acidifies the steel/corrosion product interface. In the case of galvanized steel, the corrosion rate is a function of the chloride content in the atmosphere, obtaining an excellent correlation between both parameters.     

Microbiological and biochemical changes in minimally processed fresh-cut Conference pears

Fresh-cut Conference pears were packaged under different modified atmosphere packaging (MAP) conditions and stored in refrigeration. The effects of packaging atmospheres on the microbial viability as well as on the changes in acidity, sugars and ascorbic acid (AA) were studied throughout storage. The use of plastic bags of a permeability of 15 cm ³ O ₂ m ^-2bar ^-1 24 h ^-1 and initial atmospheres of 0 kPa O ₂ extended the microbiological shelf life of pear cubes for at least 3 weeks of storage. Under these conditions, AA levels were kept almost unvaried throughout storage due to the restrictions in the O ₂ availability inside the bags' headspaces. However, these conditions also triggered physiological and biochemical changes that caused greater changes in the product acidity. The sugar profile was also affected by storage, but MAP did not show any significant influence on the reported changes.     

Distribution of isoflavones and coumestrol in neglected tropical and subtropical legumes

BACKGROUND: Isoflavones and coumestrol from dietary legumes are plant constituents showing multiple beneficial effects on humans. Owing to their ability to bind with mammalian estrogenic receptors and thereby intervention in several kinds of hormone‐related cancers, they have received much attention. Soybean (Glycine max) is currently the major source of isoflavonoids in human diet. However, dozens of tropical and subtropical leguminous species remain unexplored for their isoflavonoids content. RESULTS: We have analyzed 55 extracts from 41 tropical and subtropical legume species used either in human or animal diet by high‐performance liquid chromatography for the content of soy isoflavones, biochanin A, daidzein, daidzin, formononetin, genistein, genistin, sissotrin, ononin and the coumestan coumestrol. Genistein and biochanin A were the most abundant compounds. The highest content of genistein was found in aerial parts of Andira macrothyrsa, seeds of Pachyrhizus tuberosus and aerial parts of Calopogonium mucunoides (598, 250 and 184 µg g^?1, respectively) and biochanin A in aerial parts of Cratylia argentea, C. mucunoides and flowers of A. macrothyrsa (76, 53 and 40 µg g^?1, respectively). CONCLUSION: None of the samples tested was richer overall source of soy isoflavones and coumestrol than soybean; nevertheless several species (C. mucunoides or A. macrothyrsa) may serve as a promising source of individual compounds. © 2012 Society of Chemical Industry     

Olfactory bulb habituation to odor stimuli.

Habituation is a simple form of memory, yet its neurobiological mechanisms are only beginning to be understood in mammals. In the olfactory system, the neural correlates of habituation at a fast experimental timescale involving very short intertrial intervals (tens of seconds) have been shown to depend on synaptic adaptation in olfactory cortex. In contrast, behavioral habituation to odorants on a longer timescale with intertrial intervals of several minutes depends on processes in the olfactory bulb, as demonstrated by pharmacological studies. We here show that behavioral habituation to odorants on this longer timescale has a neuronal activity correlate in the olfactory bulb. Spiking responses of mitral cells in the rat olfactory bulb adapt to, and recover from, repeated odorant stimulation with 5-min intertrial intervals with a time course similar to that of behavioral habituation. Moreover, both the behavioral and neuronal effects of odor habituation require functioning N-methyl-d-aspartic acid receptors in the olfactory bulb. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

CTLA-4 Expression Is a Promising Biomarker of Idiopathic Pulmonary Arterial Hypertension and Allows Differentiation of the Type of Pulmonary Hypertension

Pulmonary arterial hypertension (PAH) is an increasingly frequently diagnosed disease, the molecular mechanisms of which have not been thoroughly investigated. The aim of our study was to investigate subpopulations of lymphocytes to better understand their role in the molecular pathomechanisms of various types of PAH and to find a suitable biomarker that could be useful in the differential diagnosis of PAH. Using flow cytometry, we measured the frequencies of lymphocyte subpopulations CD4+CTLA-4+, CD8+ CTLA-4+ and CD19+ CTLA-4+ in patients with different types of PAH, namely pulmonary arterial hypertension associated with congenital heart disease (CHD-PAH), pulmonary arterial hypertension associated with connective tissue disorders (CTD-PAH), chronic thromboembolic pulmonary hypertension (CTEPH) and idiopathic pulmonary arterial hypertension (iPAH), and in an age- and sex-matched control group in relation to selected clinical parameters. Patients in the iPAH group had the significantly highest percentage of CD4+CTLA-4+ T lymphocytes among all PAH groups, as compared to those in the control group (p < 0.001), patients with CTEPH (p < 0.001), CTD-PAH (p < 0.001) and CHD-PAH (p < 0.01). In iPAH patients, the percentages of CD4+CTLA-4+ T cells correlated strongly positively with the severity of heart failure New York Heart Association (NYHA) Functional Classification (r = 0.7077, p < 0.001). Moreover, the percentage of B CD19+CTLA-4+ cells strongly positively correlated with the concentration of NT-proBNP (r = 0.8498, p < 0.001). We have shown that statistically significantly higher percentages of CD4+CTLA-4+ (p ≤ 0.01) and CD8+ CTLA-4+ (p ≤ 0.001) T cells, measured at the time of iPAH diagnosis, were found in patients who died within 5 years of the diagnosis, which allows us to consider both of the above lymphocyte subpopulations as a negative prognostic/predictive factor in iPAH. CTLA-4 may be a promising biomarker of noninvasive detection of iPAH, but its role in planning the treatment strategy of PAH remains unclear. Further studies on T and B lymphocyte subsets are needed in different types of PAH to ascertain the relationships that exist between them and the disease.