Polymorphisms in the bradykinin B2 receptor gene and childhood asthma

Bradykinin has been suggested as one of the key mediators of bronchial asthma. Polymorphisms with a potential functional relevance have been described in the B2 bradykinin receptor gene. Study of these polymorphisms in 77 children with asthma and 73 controls revealed no association. However, when comparing the asthmatics according to their age at onset (before and after age 4), the exon 1 allele BE1-2G was significantly associated with late-onset asthma (p <0.05). Since BE1-2G has previously been shown to lead to a higher transcription rate of the B2 receptor, this result warrants further investigation of the role of bradykinin in conferring susceptibility to pediatric asthma

Renormalization Flow of Axion Electrodynamics

We study the renormalization flow of axion electrodynamics, concentrating on the non-perturbative running of the axion-photon coupling and the mass of the axion (like) particle. Due to a non-renormalization property of the axion-photon vertex, the renormalization flow is controlled by photon and axion anomalous dimensions. As a consequence, momentum-independent axion self-interactions are not induced by photon fluctuations. The non-perturbative flow towards the ultraviolet exhibits a Landau-pole-type behavior, implying that the system has a scale of maximum UV extension and that the renormalized axion-photon coupling in the deep infrared is bounded from above. Even though gauge invariance guarantees that photon fluctuations do not decouple in the infrared, the renormalized couplings remain finite even in the deep infrared and even for massless axions. Within our truncation, we also observe the existence of an exceptional RG trajectory, which is extendable to arbitrarily high scales, without being governed by a UV fixed point.Comment: 12 pages, 4 figure

Guanosine nucleotides regulate B2 kinin receptor affinity of agonists but not of antagonists: Discussion of a model proposing receptor precoupling to G protein

The effect of nucleotides on binding of the B2 kinin (BK) receptor agonist {[}H-3]BK and the antagonist {[}H-3]NPC17731 to particulate fractions of human foreskin fibroblasts was studied. At 0 degrees C, particulate fractions exhibited a single class of binding sites with a Kd of 2.3 nM for {[}H-3]BK and a K-d Of 3.8 nM for the antagonist {[}H-3]NPC17731. Incubation with radioligands at 37 degrees C for 5 min gave a reduction of agonist, as well as antagonist, binding that was between 0-40% depending on the preparation, even in the absence of guanosine nucleotides. As shown by Scatchard analysis, this reduction in specific binding was due to a shift in the affinity of at least a fraction of the receptors. The presence at 37 degrees C of the guanine nucleotides GTP, GDP and their poorly hydrolyzable analogs left {[}H-3]-NPC17731 binding unaffected, but reduced the receptor affinity for {[}H-3]BK to a K-d Of about 15 nM. The maximal number of receptors, however, was unchanged. This affinity change was strongly dependent on the presence of bivalent cations, in particular Mg2+. It was reversed by incubation at 0 degrees C, The rank order of the guanosine nucleotides for {[}H-3]BK binding reduction was GTP{[}gamma S] = Gpp{[}NH]p > GTP = GDP > GDP{[}beta S]. GMP, ATP, ADP and AMP showed no influence on agonist binding. A model for the interaction of the B2 kinin receptor with G proteins is discussed

Simultaneous quantitative and allele-specific expression analysis with real competitive PCR

Background: For a diploid organism such as human, the two alleles of a particular gene can be expressed at different levels due to X chromosome inactivation, gene imprinting, different local promoter activity, or mRNA stability. Recently, imbalanced allelic expression was found to be common in human and can follow Mendelian inheritance. Here we present a method that employs real competitive PCR for allele-specific expression analysis. Results: A transcribed mutation such as a single nucleotide polymorphism ( SNP) is used as the marker for allele-specific expression analysis. A synthetic mutation created in the competitor is close to a natural mutation site in the cDNA sequence. PCR is used to amplify the two cDNA sequences from the two alleles and the competitor. A base extension reaction with a mixture of ddNTPs/ dNTP is used to generate three oligonucleotides for the two cDNAs and the competitor. The three products are identified and their ratios are calculated based on their peak areas in the MALDI-TOF mass spectrum. Several examples are given to illustrate how allele-specific gene expression can be applied in different biological studies. Conclusions: This technique can quantify the absolute expression level of each individual allele of a gene with high precision and throughput

Opioid receptor activation triggering downregulation of cAMP improves effectiveness of anti-cancer drugs in treatment of glioblastoma

Glioblastoma are the most frequent and malignant human brain tumors, having a very poor prognosis. The enhanced radio- and chemoresistance of glioblastoma and the glioblastoma stem cells might be the main reason why conventional therapies fail. The second messenger cyclic AMP (cAMP) controls cell proliferation, differentiation, and apoptosis. Downregulation of cAMP sensitizes tumor cells for anti-cancer treatment. Opioid receptor agonists triggering opioid receptors can activate inhibitory Gi proteins, which, in turn, block adenylyl cyclase activity reducing cAMP. In this study, we show that downregulation of cAMP by opioid receptor activation improves the effectiveness of anti-cancer drugs in treatment of glioblastoma. The µ-opioid receptor agonist D,L-methadone sensitizes glioblastoma as well as the untreatable glioblastoma stem cells for doxorubicin-induced apoptosis and activation of apoptosis pathways by reversing deficient caspase activation and deficient downregulation of XIAP and Bcl-xL, playing critical roles in glioblastomas' resistance. Blocking opioid receptors using the opioid receptor antagonist naloxone or increasing intracellular cAMP by 3-isobutyl-1-methylxanthine (IBMX) strongly reduced opioid receptor agonist-induced sensitization for doxorubicin. In addition, the opioid receptor agonist D,L-methadone increased doxorubicin uptake and decreased doxorubicin efflux, whereas doxorubicin increased opioid receptor expression in glioblastomas. Furthermore, opioid receptor activation using D,L-methadone inhibited tumor growth significantly in vivo. Our findings suggest that opioid receptor activation triggering downregulation of cAMP is a promising strategy to inhibit tumor growth and to improve the effectiveness of anti-cancer drugs in treatment of glioblastoma and in killing glioblastoma stem cells

Initial steps for high-throughput phenotyping in vineyards

The evaluation of phenotypic characters of grapevines is required directly in vineyards and is strongly limited by time, costs and the subjectivity of person in charge. Sensor-based techniques are prerequisite in order to allow non-invasive phenotyping of individual plant traits, to increase the quantity of object records and to reduce error variation. Thus, a Prototype-Image-Acquisition-System (PIAS) was developed for semi-automated capture of geo-referenced images in an experimental vineyard. Different strategies were tested for image interpretation using MATLAB®. The interpretation of images from the vineyard with real background is more practice-oriented but requires the calculation of depth maps. Different image analysis tools were verified in order to enable contactless and non-invasive detection of bud burst and quantification of shoots at an early developmental stage (BBCH 10) and enable fast and accurate determination of the grapevine berry size at BBCH 89. Depending on the time of image acquisition at BBCH 10 up to 94 % of green shoots were visible in images. The mean berry size (BBCH 89) was recorded non-invasively with a precision of 1 mm.

Cell death sensitization of leukemia cells by opioid receptor activation

Cyclic AMP (cAMP) regulates a number of cellular processes and modulates cell death induction. cAMP levels are altered upon stimulation of specific G-protein-coupled receptors inhibiting or activating adenylyl cyclases. Opioid receptor stimulation can activate inhibitory Gi-proteins which in turn block adenylyl cyclase activity reducing cAMP. Opioids such as D,L-methadone induce cell death in leukemia cells. However, the mechanism how opioids trigger apoptosis and activate caspases in leukemia cells is not understood. In this study, we demonstrate that downregulation of cAMP induced by opioid receptor activation using the opioid D,L-methadone kills and sensitizes leukemia cells for doxorubicin treatment. Enhancing cAMP levels by blocking opioid-receptor signaling strongly reduced D,L-methadone-induced apoptosis, caspase activation and doxorubicin-sensitivity. Induction of cell death in leukemia cells by activation of opioid receptors using the opioid D,L-methadone depends on critical levels of opioid receptor expression on the cell surface. Doxorubicin increased opioid receptor expression in leukemia cells. In addition, the opioid D,L-methadone increased doxorubicin uptake and decreased doxorubicin efflux in leukemia cells, suggesting that the opioid D,L-methadone as well as doxorubicin mutually increase their cytotoxic potential. Furthermore, we found that opioid receptor activation using D,L-methadone alone or in addition to doxorubicin inhibits tumor growth significantly in vivo. These results demonstrate that opioid receptor activation via triggering the downregulation of cAMP induces apoptosis, activates caspases and sensitizes leukemia cells for doxorubicin treatment. Hence, opioid receptor activation seems to be a promising strategy to improve anticancer therapies