Spinal Excitatory Dynorphinergic Interneurons Contribute to Burn Injury-Induced Nociception Mediated by Phosphorylated Histone 3 at Serine 10 in Rodents

The phosphorylation of serine 10 in histone 3 (p-S10H3) has recently been demonstrated to participate in spinal nociceptive processing. However, superficial dorsal horn (SDH) neurons involved in p-S10H3-mediated nociception have not been fully characterized. In the present work, we combined immunohistochemistry, in situ hybridization with the retrograde labeling of projection neurons to reveal the subset of dorsal horn neurons presenting an elevated level of p-S10H3 in response to noxious heat (60 °C), causing burn injury. Projection neurons only represented a small percentage (5%) of p-S10H3-positive cells, while the greater part of them belonged to excitatory SDH interneurons. The combined immunolabeling of p-S10H3 with markers of already established interneuronal classes of the SDH revealed that the largest subset of neurons with burn injury-induced p-S10H3 expression was dynorphin immunopositive in mice. Furthermore, the majority of p-S10H3-expressing dynorphinergic neurons proved to be excitatory, as they lacked Pax-2 and showed Lmx1b-immunopositivity. Thus, we showed that neurochemically heterogeneous SDH neurons exhibit the upregulation of p-S10H3 shortly after noxious heat-induced burn injury and consequential tissue damage, and that a dedicated subset of excitatory dynorphinergic neurons is likely a key player in the development of central sensitization via the p-S10H3 mediated pathway.     

Aspects of fetal physiology from 18 to 37 weeks' gestation as assessed by blood sampling

OBJECTIVE: To construct reference ranges for fetal pH, oxygen pressure (PO2), and hematologic and biochemical blood constituents, which can be used to analyze changes with gestation and differences with maternal values, thus elucidating some aspects of fetal biology and the effects of the maternal and placental environments. METHODS: We assayed venous pH, PO2, hematocrit, glucose, uric acid, urea, creatinine, total protein, total and direct bilirubin, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, alkaline phosphatase, lactic dehydrogenase, amylase, pseudocholinesterase, creatine kinase, triglycerides, and cholesterol concentrations in 157 fetuses and 134 mothers who underwent fetal blood sampling from 18 to 37 weeks' gestation. None of the fetuses was infected or had chromosomal, hematologic, or hormonal abnormalities. RESULTS: All the variables analyzed were similar in fetuses sampled at the placental cord insertion (n = 125) or at the intrahepatic vein (n = 32). Maternal and fetal concentrations of glucose (r = 0.79, P < .001), urea (r = 0.96, P < .001), creatinine (r = 0.83, P < .001), and uric acid (r = 0.94, P < .001) correlated significantly, and their differences exhibited significant changes: the maternal-fetal differences of glucose and urea increased, whereas those of uric acid and creatinine decreased with advancing gestation. Fetal pH and PO2 decreased with gestational age, whereas hematocrit increased, similar to what has been described previously. All of the other variables, with the exception of amylase and cholesterol, changed significantly during the investigated period of pregnancy. Gestational age explained at least 40% of the variance in values of fetal total protein, pseudocholinesterase, alanine aminotransferase, creatine kinase, and triglycerides, but only 3-25% of the variation in the remainder. Most enzymes were higher in the fetus than in the maternal circulation, and all except alkaline phosphatase increased with gestational age. The maternal-fetal glucose difference correlated significantly with hematocrit, pH, and PO2, independent of gestational age and independent of each other. CONCLUSION: With the exception of aspartate aminotransferase, all of the analyzed fetal variables were different from the maternal values, and most changed with gestational age. The mechanisms leading to these fetal specificities remain mostly uncertain, but the provision of reference ranges for several blood constituents may be useful in the differential diagnosis of fetal disease.     

Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH,FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques

Metabolic FLIM (fluorescence lifetime imaging) is used to image bioenergetic status in cells and tissue. Whereas an attribution of the fluorescence lifetime of coenzymes as an indicator for cell metabolism is mainly accepted, it is debated whether this is valid for the redox state of cells. In this regard, an innovative algorithm using the lifetime characteristics of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) to calculate the fluorescence lifetime induced redox ratio (FLIRR) has been reported so far. We extended the FLIRR approach and present new results, which includes FLIM data of the various enzymes, such as NAD(P)H, FAD, as well as flavin mononucleotide (FMN). Our algorithm uses a two-exponential fitting procedure for the NAD(P)H autofluorescence and a three-exponential fit of the flavin signal. By extending the FLIRR approach, we introduced FLIRR1 as protein-bound NAD(P)H related to protein-bound FAD, FLIRR2 as protein-bound NAD(P)H related to free (unbound) FAD and FLIRR3 as protein-bound NAD(P)H related to protein-bound FMN. We compared the significance of extended FLIRR to the metabolic index, defined as the ratio of protein-bound NAD(P)H to free NAD(P)H. The statistically significant difference for tumor and normal cells was found to be highest for FLIRR1.     

Synthesis,Characterization and Application of Sucrose Carboxylic Acid Polyesters — A Prototype of Non-Caloric or Surface Active Compounds

Sucrose carboxylic acid polyesters are in dependence of the degree of acylation and fatty acid composition more or less undigestible or emulsifying fat substitutes and functional additives for foods. In this connection the relationships between their molecular parameters and functional properties are discussed. New procedures of the solvent-free synthesis are presented. Furthermore, open biochemical and toxicological problems of pinocytosis and accumulation of intact polyesters in organs and tissue being still under investigations are considered.     

Knowledge transfer in IT offshoring relationships: the roles of social capital,efficacy and outcome expectations

Information technology (IT) development in global organisations relies heavily on the transfer of tacit and complex knowledge from onshore units to offshore subsidiaries. A central concern of such organisations is the development of social capital, which is known to facilitate the smooth transfer of knowledge. However, only a few studies in IS research have explicitly examined the role of social capital for knowledge transfer in an IT offshoring context. In this paper, we argue that such knowledge transfer mechanisms can be understood better by considering social capital in concert with knowledge senders' efficacy and outcome expectations, two of the potentially key motivational drivers of knowledge transfer. We develop our arguments through a qualitative case study of a large German multinational company. German IT developers in this firm provided in‐depth accounts of their experience with offshore colleagues in an Indian captive subsidiary unit. Drawing on our analysis, we develop a model that depicts the influence of social capital, efficacy and outcome expectations on onshore IT developers' ability and willingness to transfer knowledge to offshore colleagues. Through the model, we also explain how social capital, efficacy and outcome expectations are interrelated and generate three interlocked, self‐reinforcing circles of knowledge transfer success in IT offshoring relationships.     

Zum Verbesserungspotenzial schriftlicher Aufklärungsmaterialien zu (bio)medizinischen Forschungsvorhaben – Empirische Analyse von Antragsunterlagen einer Forschungsethikkommission

Background

One of the elementary prerequisites for medical research on and with humans is the patients’ or probands’ informed consent. To ensure informed consent, study participants must be—among other things—provided with high-quality information. We developed criteria to assess and evaluate the quality of various written patient information material.

Methods

Based on a catalogue addressing 117 single criteria, we assessed the quality of 128 randomly selected documents from study proposals submitted to the ethical committee of Luebeck University in 2006. Each criterion refers to one of six quality areas (such as “readability and comprehensibility” or “potential benefit and harm”).

Results

The documents on average satisfied half of the criteria with a range from 20–76% for single items. The area with the highest quality score was “consent form” (64%), while “potential benefit and harm” (35%) was the lowest. Material from drug trials showed a significantly higher quality than that from other study types. Only 21 out of 117 criteria were met in more than 80% of all relevant documents.

Conclusion

The study provides evidence for significant deficits in the information material from basic and clinical research projects presented to one academic research ethics committee. Researchers need support in developing and writing informed consent documents. Our set of criteria could be used to make them more sensitive to the various demands involved.     

SDS electrophoresis of legume seed proteins in horizontal ultrathin-layer pore gradient gels

Summary Horizontal SDS electrophoresis of 18 legume seed protein extracts was performed in ultrathin-layer polyacrylamide gels on foil supports. Separation results of the SD S pore gradient electrophoresis (T=4–22.5%) are compared to those of SDS electrophoresis in a constant pore size gel (T=10%). Resolution as well as the sensitivity (0.1 g protein per band) of the ultrathin-layer SDS pore-gradient electrophoresis were extremely high. Because of the very low gel thickness, separation, staining and drying were completed in substantially shorter times than achieved with conventional thick gels. An easy technique for casting ultrathin-layer (360 gm) concave gradient gels for 10 cm separation distance and a width of 25 cm is described. The even distribution of the concave exponential pore-gradient over the whole gel width is demonstrated. Molecular weights of the legume proteins are detected from 5,000 to 110,000 daltons. The protein patterns are genus- and species-specific.

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Horizontale Ultradünnschicht SDS-Gradientengel-Elektrophorese von Leguminosensamenproteinen

Zusammenfassung Die ausgezeichnete Trennschärfe der horizontalen Ultradünnschicht-SDS-Gradienten-gel Elektrophorese wird am Beispiel von Samenproteinen 18 verschiedener Leguminosengattungen, -arten und -sorten gezeigt. Es werden die Trennergebnisse der SDS-Elektrophorese mit Gelgradienten (T=4-22,5%) bzw. mit Gelen konstanter Porengröße (T=10%) verglichen. Das höchste Auslösungsvermögen und die beste Trennschärfe zeigen ultradünne Gradientengele. Die Nachweisempfindlichkeit ist bei allen Ultradünn-schicht-SDS-Elektrophoresen sehr hoch (0,1 g Protein/Bande). Da die auf Folie polymerisierten Gele sehr dünn sind (360 m) kann mit wesentlich verkürzten Trenn-, Färbe-, Entfärbe- und Trocknungszeiten gear-beitet werden. Es wird eine einfache Herstellung ultradünner Polyacrylamidgele mit exponentiellen konkaven Gradienten für die Trenndistanz von 10 cm mit einer Breite von 25 cm beschrieben. Die gerade und gleichmäßige Verteilung des Gradienten über die gesamte Gelbreite wird gezeigt. Mit der beschriebenen Methode werden bei den untersuchten Leguminosen-proteinen Molekulargewichte von 5 000 his 110 000 Dalton gefunden. Die Proteinmuster erweisen sich als gattungs- und artspezifisch.

     

Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus

Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single-chain F_v (scF_v), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scF_v were derived from three different human V_H germline genes. The binding specificities were confirmed by Western blots and ELISA. The scF_v preparations reacted with B. napus ENR, but not with β-keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scF_v 3.13 recognizes an epitope located within the n-terminal 80 amino acids of the enzyme molecule. The scF_v were used to detect ENR directly in extracts of B. napus seeds.