6. SHORT NOTES

Schmitt, M. B. 1975. Observations on the Black Crake in the Southern Transvaal. Ostrich 46:129-138.

During a four-year study period, several pairs of Black Crakes Porzana flavirostris and their progeny have been studied on a vlei in the southern Transvaal. Morphological differences between male and female are discussed, a double-brooded breeding season is indicated, the development of chicks is described, and the moult cycle is indicated. The Black Crake moults all flight feathers simultaneously and during its breeding season.     

Doping control analysis of trenbolone and related compounds using liquid chromatography-tandem mass spectrometry

Trenbolone (17β-hydroxy-estra-4,9,11-trien-3-one) and its derivatives such as 17α-methyltrenbolone represent a class of highly potent anabolic-androgenic steroids, which are prohibited in sports according to the regulation of the World Anti-Doping Agency (WADA). Due to marginal gas chromatographic properties of these compounds but excellent proton affinities resulting from a large and conjugated π-electron system, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been the method of choice for the detection of these analytes in sports drug testing. Recent findings of trenbolone and methyltrenbolone in doping control urine samples of elite athletes demonstrated the importance of a sensitive and robust analytical method, which was based on an enzymatic hydrolysis of target compounds, liquid-liquid extraction and subsequent LC-MS/MS measurement. Diagnostic product ions obtained after collision-induced dissociation of protonated molecules were found at m/z 227, 211, 199 and 198, which enabled targeted screening using multiple reaction monitoring. Using 7 model compounds (trenbolone, epitrenbolone, methyltrenbolone, ethyltrenbolone, propyltrenbolone, 17-ketotrenbolone and altrenogest), the established method was validated for specificity, lower limits of detection (0.3-3 ng/mL), recovery (72-105%), intraday and interday precision (≤20%).     

A Risk Prediction Model for the Assessment and Triage of Women with Hypertensive Disorders of Pregnancy in Low-Resourced Settings: The miniPIERS (Pre-eclampsia Integrated Estimate of RiSk) Multi-country Prospective Cohort Study

Background

Pre-eclampsia/eclampsia are leading causes of maternal mortality and morbidity, particularly in low- and middle- income countries (LMICs). We developed the miniPIERS risk prediction model to provide a simple, evidence-based tool to identify pregnant women in LMICs at increased risk of death or major hypertensive-related complications.

Methods and Findings

From 1 July 2008 to 31 March 2012, in five LMICs, data were collected prospectively on 2,081 women with any hypertensive disorder of pregnancy admitted to a participating centre. Candidate predictors collected within 24 hours of admission were entered into a step-wise backward elimination logistic regression model to predict a composite adverse maternal outcome within 48 hours of admission. Model internal validation was accomplished by bootstrapping and external validation was completed using data from 1,300 women in the Pre-eclampsia Integrated Estimate of RiSk (fullPIERS) dataset. Predictive performance was assessed for calibration, discrimination, and stratification capacity. The final miniPIERS model included: parity (nulliparous versus multiparous); gestational age on admission; headache/visual disturbances; chest pain/dyspnoea; vaginal bleeding with abdominal pain; systolic blood pressure; and dipstick proteinuria. The miniPIERS model was well-calibrated and had an area under the receiver operating characteristic curve (AUC ROC) of 0.768 (95% CI 0.735–0.801) with an average optimism of 0.037. External validation AUC ROC was 0.713 (95% CI 0.658–0.768). A predicted probability ≥25% to define a positive test classified women with 85.5% accuracy. Limitations of this study include the composite outcome and the broad inclusion criteria of any hypertensive disorder of pregnancy. This broad approach was used to optimize model generalizability.

Conclusions

The miniPIERS model shows reasonable ability to identify women at increased risk of adverse maternal outcomes associated with the hypertensive disorders of pregnancy. It could be used in LMICs to identify women who would benefit most from interventions such as magnesium sulphate, antihypertensives, or transportation to a higher level of care. Please see later in the article for the Editors'' Summary     

Estimation of phylogenetic relationships within the Ascomycota on the basis of 18S rDNA sequences and chemotaxonomy

Small subunit rRNA gene sequences (18S rDNA), cell wall carbohydrate composition and ubiquinone components were analysed within a larger number of ascomycetous yeasts and dimorphic fungi to validate their congruence in predicting phylogenetic relationships. The glucose-mannose pattern distinguishes the Hemiascomycetes from the Euascomycetes and the Protomycetes which are characterised with the glucose-mannose-galactose-rhamnose-(fucose) profile. The glucose-mannose-galactose pattern was found in the cell walls of all the three classes. Different coenzyme Q component (CoQ5 to CoQ10) were found within the representatives of the Hemiascomycetes. Whereas CoQ9, CoQ10 and CoQ10H2 predominate within the Euascomycetes, CoQ9 and CoQ10 characterise the Protomycetes. Chemotaxonomic studies coupled with additional molecular and co-evolution studies support the idea that the Hemiascomycetes occupy a basal position in the phylogeny of Ascomycota. These results are not in line with the phylogenetic studies based on the sequences of 18S rRNA encoding gene. The maximum parsimony analysis indicated that Hemiascomycetes and Protomycetes might represent sister groups, opposing to the earlier reported results, where the Archiascomycetes (Protomycetes) or the Hemiascomycetes had been considered to be the most primitive ascomycetous fungi. Instead of the class Archiascomycetes, the term Protomycetes was introduced reflecting much better the properties of the whole class.     

Mechanistic studies of sesquiterpene cyclases based on their carbon isotope ratios at natural abundance

During the process of terpene biosynthesis, C–C bond breaking and forming steps are subjected to kinetic carbon isotope effects, leading to distinct carbon isotopic signatures of the products. Accordingly, carbon isotopic signatures could be used to reveal the ‘biosynthetic history’ of the produced terpenoids. Five known sesquiterpene cyclases, regulating three different pathways, representing simple to complex biosynthetic sequences, were heterologously expressed and used for in vitro assays with farnesyl diphosphate as substrate. Compound specific isotope ratio mass spectrometry measurements of the enzyme substrate farnesyl diphosphate (FDP) and the products of all the five cyclases were performed. The calculated δ¹³C value for FDP, based on δ¹³C values and relative amounts of the products, was identical with its measured δ¹³C value, confirming the reliability of the approach and the precision of measurements. The different carbon isotope ratios of the products reflect the complexity of their structure and are correlated with the frequency of carbon–carbon bond forming and breaking steps on their individual biosynthetic pathways. Thus, the analysis of carbon isotopic signatures of terpenes at natural abundance can be used as a powerful tool in elucidation of associated biosynthetic mechanisms of terpene synthases and in future in vivo studies even without ‘touching’ the plant.     

Storage‐induced changes of the cytosolic red blood cell proteome analyzed by 2D DIGE and high‐resolution/high‐accuracy MS

The storage of packed red blood cells (RBCs) is associated with the development of morphological and biochemical changes leading to a reduced posttransfusion functionality and viability of the cells. Within this study, 2D DIGE and high‐resolution/high‐accuracy Orbitrap MS were used to analyze the storage‐induced changes of the cytosolic RBC proteome and identify characteristic protein patterns and potential marker proteins for the assessment of RBC storage lesions. Leukodepleted RBC concentrates were stored according to standard blood bank conditions for 0, 7, 14, 28, and 42 days and analyzed by using a characterized and validated protocol. Following statistical evaluation, a total of 14 protein spots were found to be significantly altered after 42 days of ex vivo storage. Protein identification was accomplished by tryptic digestion and LC‐MS/MS and three proteins potentially useful as biomarkers for RBC aging comprising transglutaminase 2, beta actin, and copper chaperone for superoxide dismutase were selected and validated by western blot analysis. These can serve as a basis for the development of a screening assay to detect RBC storage lesions and autologous blood doping in sports.     

Chemistry and geographic variation of floral scent in Yucca filamentosa (Agavaceae)

We identified volatiles from the floral headspace of Yucca filamentosa using gas chromatography and mass spectrometry and analyzed floral scent composition and variation among populations pollinated by different yucca moth species. Twenty-one scent compounds were repeatedly identified and most could be categorized into two major classes: (1) homoterpenes derived from the sesquiterpene alcohol nerolidol and (2) long chain aliphatic hydrocarbons. Two biosynthetic pathways are thus responsible for the majority of floral volatiles in Y. filamentosa. The homoterpene E-4,8-dimethylnona-1,3,7-triene, which is released systemically by higher plants upon herbivory, was the most abundant compound. Two di-oxygenated compounds not previously reported as floral compounds also were detected. No differentiation in floral scent was observed between populations pollinated by different yucca moths, nor was there any correlation between chemical distance and geographic distance among populations. The total release rate of volatiles differed significantly among populations, but not between populations with different pollinators. The combination of unique compounds and low variation in the fragrance blend may reflect highly selective attraction of obligate pollinators to flowers. The observed lack of differentiation in floral scent can putatively explain high moth-mediated gene flow among sites, but it does not explain conservation of odor composition across populations with different pollinators.