Glucosinolate profiles and phylogeny in Barbarea compared to other tribe Cardamineae (Brassicaceae) and Reseda (Resedaceae), based on a library of ion trap HPLC-MS/MS data of reference desulfoglucosinolates.

A library of ion trap MS2 spectra and HPLC retention times reported here allowed distinction in plants of at least 70 known glucosinolates (GSLs) and some additional proposed GSLs. We determined GSL profiles of selected members of the tribe Cardamineae (Brassicaceae) as well as Reseda (Resedaceae) used as outgroup in evolutionary studies. We included several accessions of each species and a range of organs, and paid attention to minor peaks and GSLs not detected. In this way, we obtained GSL profiles of Barbarea australis, Barbarea grayi, Planodes virginica selected for its apparent intermediacy between Barbarea and the remaining tribe and family, and Rorippa sylvestris and Nasturtium officinale, for which the presence of acyl derivatives of GSLs was previously untested. We also screened Armoracia rusticana, with a remarkably diverse GSL profile, the emerging model species Cardamine hirsuta, for which we discovered a GSL polymorphism, and Reseda luteola and Reseda odorata. The potential for aliphatic GSL biosynthesis in Barbarea vulgaris was of interest, and we subjected P-type and G-type B. vulgaris to several induction regimes in an attempt to induce aliphatic GSL. However, aliphatic GSLs were not detected in any of the B. vulgaris types. We characterized the investigated chemotypes phylogenetically, based on nuclear rDNA internal transcribed spacer (ITS) sequences, in order to understand their relation to the species B. vulgaris in general, and found them to be representative of the species as it occurs in Europe, as far as documented in available ITS-sequence repositories. In short, we provide GSL profiles of a wide variety of tribe Cardamineae plants and conclude aliphatic GSLs to be absent or below our limit of detection in two major evolutionary lines of B. vulgaris. Concerning analytical chemistry, we conclude that availability of authentic reference compounds or reference materials is critical for reliable GSL analysis and characterize two publicly available reference materials: seeds of P. virginica and N. officinale.

Multiple hydroxyphenethyl glucosinolate isomers and their tandem mass spectrometric distinction in a geographically structured polymorphism in the crucifer Barbarea vulgaris.

Two distinct glucosinolate (GSL) chemotypes (P and G-types) of Barbarea vulgaris (Brassicaceae) were known from southern Scandinavia, but whether the types were consistent in a wider geographic area was not known. Populations (26) from Eastern and Central Europe were analyzed for GSLs in order to investigate whether the two types were consistent in this area. Most (21) could be attributed to one of the previously described GSL profile types, the P-type (13 populations) and the G-type (8 populations), based on differences in the stereochemistry of 2-hydroxylation, presence or absence of phenolic glucobarbarin derivatives, and qualitative differences in indole GSL decoration (tested for a subset of 8+6 populations only). The distinction agreed with previous molecular genetic analysis of the same individuals. Geographically, the P-type typically occurred in Eastern Europe while the G-type mainly occurred in Central Europe. Of the remaining five populations, minor deviations were observed in some individuals from two populations genetically assigned to the G-type, and a hybrid population from Finland contained an additional dihydroxyphenethyl GSL isomer attributed to a combinatorial effect of P-type and G-type genes. Major exceptions to the typical GSL profiles were observed in two populations: (1) A G-type population from Slovenia deviated by a high frequency of a known variant in glucobarbarin biosynthesis ('NAS form') co-occurring with usual G-type individuals. (2) A population from Caucasus exhibited a highly deviating GSL profile dominated by p-hydroxyphenethyl GSL that was insignificant in other accessions, as well as two GSLs investigated by NMR, m-hydroxyphenethylGSL and a partially identified m,p disubstituted hydroxy-methoxy derivative of phenethylGSL. Tandem HPLC-MS of seven NMR-identified desulfoGSLs was carried out and interpreted for increased certainty in peak identification and as a tool for partial structure elucidation. The distinct, geographically separated chemotypes and rare variants are discussed in relation to future taxonomic revision and the genetics and ecology of GSLs in B. vulgaris.

Different geographical distributions of two chemotypes of Barbarea vulgaris that differ in resistance to insects and a pathogen.

The interactions of plants with herbivores and pathogens have been suggested to drive the evolution of resistances in plants and in some cases new lineages and taxa. However, such divergence may require reproductive isolation, e.g., in allopatry. In the crucifer Barbarea vulgaris, some plants are resistant to the flea beetle Phyllotreta nemorum, due to production of specific saponins, whereas others are susceptible. Resistant and susceptible plants additionally differ in resistance to the pathogen Albugo candida, content of glucosinolates, and leaf pubescence, and they are genetically strongly divergent and partially reproductively incompatible. This suggests that at some point they were separated for a considerable length of time. Previously, the insect susceptible P-type had been described only from Denmark, Sweden, and Estonia, whereas the resistant G-type is widely distributed in Western Europe. Here, we tested whether the two plant types have divergent geographical distributions and maintain their distinct trait associations throughout their range. The insect-susceptible type was found in Russia, the Baltics, and parts of Fennoscandia, but not in Central Europe. In contrast, the insect resistant type was found from Finland and westwards. Their different trait associations were consistent within the two ranges. We therefore suggest that the two plant types diverged in allopatry at some time in the past, and evolved different resistances in response to local antagonists. The two plant types probably maintain their distinctness due to a hybridization barrier. Thus, the present distributions of the two types may be shaped by both historical processes and current differential biotic selection.

Practice patterns for the surgical treatment of T1 renal cell carcinoma: a nationwide population-based register study.

OBJECTIVE: Treatment of renal cell carcinoma (RCC) with radical nephrectomy (RN) induces chronic kidney disease more frequently compared with nephron-sparing surgery (NSS), which may have an impact on overall survival. Thus, NSS is recommended for RCCs up to 7 cm (T1). The aim of this study was to determine the extent to which these recommendations are implemented in clinical practice. MATERIAL AND METHODS: Data were extracted from the Swedish National Kidney Cancer Register, a population-based register covering 98% of all RCCs in Sweden. In total, 3158 patients (1892 men, 1266 women) were primarily diagnosed with cT1N0M0 and treated surgically during 2005-2011. The administered treatments were evaluated between different hospitals as well as between the 21 independent healthcare counties. RESULTS: In all, 742 patients were treated with NSS, 2339 with RN and 77 with minimally invasive ablative treatments. For cT1a RCC, patients treated with NSS increased from 22% in 2005 to 53% in 2011, and for cT1b from 2% to 10%. Nephron-sparing treatments for cT1a RCC were performed in 62% in university hospitals, 34% in intermediate- and 11% in low-volume hospitals. There was significant (p < 0.001) variation (31-67%) between the university hospitals and also for patient care in the 21 different counties (16-78%). There was an increased relative survival after NSS for T1a patients compared with RN. The register design by itself indicates limitations using data gathered from all Swedish hospitals. CONCLUSIONS: NSS was underutilized in many hospitals and a patient's chance of being offered NSS varied according to their place of residence. Patients with cT1a RCC treated with NSS had a significantly better relative survival than those treated with RN.

Preliminary studies of the physical stability of a glucagon-like peptide-1 derivate in the presence of metal ions.

The physical stability and the secondary structure of a glucagon-like peptide-1 derivative were investigated in the presence of the metal ions Al(3+), Zn(2+), Mg(2+), and K(+), known as possible leachables from container-closure systems. Metal ions were investigated in concentrations of 0-50 ppm. Test solutions of the peptide were exposed to elevated temperature (25 degrees C) and rotation (37 degrees C) for up to 4 weeks. The samples were examined by nephelometry, thioflavine T fluorescence, and Fourier-transform infrared spectroscopy. Readily prepared test solutions were examined by tryptophan fluorescence. The stability profiles were unchanged after addition of Mg(2+) and K(+) in 0-50 ppm concentrations. However, a concentration-dependent increase in thioflavine intensities was observed after addition of Al(3+) and Zn(2+). The destabilising effect of Al(3+) and Zn(2+) was furthermore confirmed by FTIR as the secondary structure of the peptide changed from predominantly alpha-helix to a higher beta-sheet content. Additionally Al(3+) changed the secondary structure of the peptide using Trp fluorescence.