Multi‐surveyor capture‐mark‐recapture as a powerful tool for butterfly population monitoring in the pre‐imaginal stage

For many elusive insect species, which are difficult to cover by standard monitoring schemes, innovative survey methods are needed to gain robust data on abundance and population trends. We suggest a monitoring of overwintering larvae for the endangered nymphalid butterfly Limenitis reducta. We tested different removal and capture‐mark‐recapture (CMR) approaches in a field study in the “Alb‐Donau” region, Germany. Classical removal and CMR studies require movement of the organisms under study, but in our approach, we replaced movement of the study organisms by random movement of multiple different surveyors. We tested the validity of the approach by comparing detection frequencies from our field data with simulated detections. Our results indicate that multi‐surveyor removal/CMR techniques are suitable for estimating abundance of overwintering L. reducta larvae. Depending on surveyor experience, the average detection probability ranged between 16% for novices and 35% for experts. The uncertainty of population estimates increased with a decrease in personnel expenditure. Estimated larval densities on a spruce clear‐cut varied between one and three individuals per 100 m², probably related to habitat conditions. We suggest a CMR approach with three to four trained surveyors for the monitoring of L. reducta populations in the overwintering stage. Compared with previous sampling methods, our approach is a powerful tool with clear advantages: long survey period, estimates of the absolute population size accompanied by uncertainty measures, and estimates of overwinter mortality. The proposed method can be adapted and used for several different butterfly species, other insect taxa with specific immobile life stages, and some sessile organisms, for example, elusive plants, fungi, or corals.     

Isolation and characterisation of genes for sulphate activation and reduction in Aspergillus nidulans: implications for evolution of an allosteric control region by gene duplication

A region of the Aspergillus nidulans genome carrying the sA and sC genes, encoding PAPS reductase and ATP sulphurylase, respectively, was isolated by transformation of an sA mutant with a cosmid library. The genes were subcloned and their functions confirmed by retransformation and complementation of A. nidulans strains carrying sA and sC mutations. The physical distance of 2 kb between the genes corresponds to a genetic distance of 1 cM. While the deduced amino acid sequence of the sA gene product shows homology with the equivalent MET16 gene product of Saccharomyces cerevisiae, the sC gene product resembles the equivalent MET3 yeast gene product at the N-terminal end, but differs markedly from it at the C-terminal end, showing homology to the APS kinases of several microorganisms. It is proposed that this C-terminal region does not encode a functional APS kinase, but is responsible for allosteric regulation by PAPS of the sulphate assimilation pathway in A. nidulans, and that the ATP sulphurylase encoding-gene (sC) of filamentous ascomycetes may have evolved from a bifunctional gene similar to the nodQ gene of Rhizobium meliloti.     

Dysregulation of dopamine-dependent mechanisms as a determinant of hypertension: studies in dopamine receptor knockout mice

Dopamine plays an important role in the pathogenesis of hypertension by regulating epithelial sodium transport and by interacting with vasoactive hormones/humoral factors, such as aldosterone, angiotensin, catecholamines, endothelin, oxytocin, prolactin pro-opiomelancortin, reactive oxygen species, renin, and vasopressin. Dopamine receptors are classified into D(1)-like (D(1) and D(5)) and D(2)-like (D(2), D(3), and D(4)) subtypes based on their structure and pharmacology. In recent years, mice deficient in one or more of the five dopamine receptor subtypes have been generated, leading to a better understanding of the physiological role of each of the dopamine receptor subtypes. This review summarizes the results from studies of various dopamine receptor mutant mice on the role of individual dopamine receptor subtypes and their interactions with other G protein-coupled receptors in the regulation of blood pressure.     

Improvement of ethyl glucuronide determination in human urine and serum samples by solid-phase extraction

An improved method for the determination of ethyl glucuronide (EtG) in human serum and urine was developed using solid-phase extraction (SPE) and gas chromatography (GC) with mass spectrometric detection (MS). EtG was isolated from serum and urine using aminopropyl SPE columns after deproteination with perchloric acid and hydrochloric acid, respectively. The chromatographic separation was performed on a DB 1701 fused-silica column. At a signal-to-noise ratio of 3:1, a quantification limit of 173 and 560 ng/ml and a detection limit of 37 and 168 ng/ml could be determined for serum and urine, respectively. This indicates high specificity and sensitivity of the described method. The mean absolute recovery was 85%, while intra- and inter-day precision of the assay were all less than 7.5%. The linearity of the calibration curves was satisfying as indicated by correlation coefficients of >0.993. The presented method provides the basis for determination and identification of EtG in human serum and urine samples in a low-concentration range for monitoring alcohol consumption during treatment for alcohol dependence and comorbid alcohol abuse of psychotherapy patients.