Nitric oxide detection by DAF (diaminofluorescein) fluorescence in human myometrial tissue.

Nitric oxide (NO) is considered to be involved in the modulation of uterine contractility. In the present pilot study, the direct detection of intracellular NO in pregnant human myometrial tissues has been investigated by using the fluorescent indicator 4,5-diaminofluorescein-2 diacetate (DAF-2DA). Pregnant myometrial tissue samples were obtained during Cesarean sections between week 34 and 40 of gestation before the onset of labor.

Butterfly wings, a new site of porphyrin synthesis and cleavage: studies on the expression of the lipocalin bilin-binding protein in Pieris brassicae.

The bilin-binding protein (BBP), a member of the lipocalin protein superfamily, is synthesized mainly in last instar larvae and in late pupae and newly emerged adults of Pieris brassicae, as previously reported. Here we present results from Northern blot analysis of the BBP gene transcript and from in vitro studies of holo-BBP biosynthesis with isolated wings using [14C]5-aminolevulinic acid as a precursor to the bilin ligand, [14C]-amino acids to label the apo-protein and inhibitors for both processes. Our combined data clearly demonstrate that BBP, which accumulates around pupa-adult transformation, is produced as holoprotein in the developing wings, while the BBP gene transcript is no longer detected in the rest of the body.

Evaluation of different strategies for real-time RT-PCR expression analysis of corticotropin-releasing hormone and related proteins in human gestational tissues.

In this article real-time quantitative RT-PCR strategies for investigation of mRNA distribution in stress-related corticotropin-releasing hormone (CRH), CRH-binding protein (CRH-BP), and CRH receptors (CRH-R1, CRH-R2) in human gestational tissues are described. The effect of sample and RNA preparation, reverse transcription, and the quantitation strategy were investigated. Both thawing of the sample before homogenization and the RT reaction were identified as critical steps.