On the evolution of chloroplasts from eukaryotic endosymbionts: A study of Glenodinium foliaceum (Dinophyceae)

A study of the marine dinoflagellate Glenodinium foliaceum is presented which attempts to provide information on the process of host-endosymbiont integration in a unicellular system that may be of relevance to current hypotheses on chloroplast evolution. An experimental ultrastructural review of G. foliaceum supports the proposal that the chloroplasts and supernumerary nucleus belong to a morphologically reduced, endosymbiotic chromophyte. DAPI staining shows that the endosymbiont nucleus is fragmented in some cells and appears to be randomly divided by the host's cleavage furrow at cytokinesis. A quantification of the intensity of mithramycin fluorescence from single cells by flow cytometry suggests that both nuclei synthesize their ENA in synchrony. Protocols are outlined for isolating the chloroplasts and dinoflagellate nucleus from G. foliaceum and a method is given for fractionating whole cell lysates to prepare chloroplast, host nuclear, and endosymbiont nuclear DNAs. It is estimated that the symbiont and dinoflagellate nuclei contain about 34 pg and 40 pg of ENA respectively. There is no evidence of amplified genesized ENA molecules in the symbiont nucleus. The results of preliminary re-association experiments show only the presence of highly repeated ENA and sequences of intermediate repetitiveness in total cell ENA, which together comprise about half the genome. The chloroplast DNA is shown to a molecular weight of approximately 103 kb by restriction endonuclease analysis but an attempt to localize the rbc S gene in either the chloroplast or nuclear DNAs using a cloned pea ENA probe proved unsuccessful. Two other anomalously pigmented dinoflagellates, Gyrodinium aureolum and Gymnodinium galatheanum, are demonstrated to lack endosymbionts by electron and fluorescence microscopy, but the atypical ultrastructural or cytological characters of their chloroplasts are considered to indicate an endosymbiotic origin. The possibility that Glenodinium foliaceum could evolve into a similar uninucleate species is discussed in relation to the conservation of genetic information for nuclear-encoded, chloro-plast proteins. <p

Variation of theanine, phenolic, and methylxanthine compounds in 21 cultivars of Camellia sinensis harvested in different seasons

This is the first study to use chemometric methods to differentiate among 21 cultivars of Camellia sinensis from China and between leaves harvested at different times of the year using 30 compounds implicated in the taste and quality of tea. Unique patterns of catechin derivatives were observed among cultivars and across harvest seasons. C. sinensis var. pubilimba (You 510) differed from the cultivars of C. sinensis var. sinensis, with higher levels of theobromine, (+)-catechin, gallocatechin, gallocatechin gallate and theasinensin B, and lower levels of (−)-epicatechin, (−)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG), respectively. Three cultivars of C. sinensis var. sinensis, Fuyun 7, Qiancha 7 and Zijuan contained significantly more caffeoylquinic acids than others cultivars. A Linear Discriminant Analysis model based on the abundance of 12 compounds was able to discriminate amongst all 21 tea cultivars. Harvest time impacted the abundance of EGC, theanine and afzelechin gallate

Is aristolochic acid nephropathy a widespread problem in developing countries? A case study of Aristolochia indica L. in Bangladesh using an ethnobotanical - phytochemical approach

Ethnopharmacological relevance: Species of Aristolochia are associated with aristolochic acid nephropathy (AAN), a renal interstitial fibrosis and upper urinary tract cancer (UUC). Aristolochic acid nephropathy has been reported in ten countries but its true incidence is unknown and most likely underestimated. By combining an ethnobotanical and phytochemical approach we provide evidence for the risk of AAN occurring in Bangladesh. More specifically, we assess the intra-specific variation of aristolochic acid analogues in medicinally used A. indica samples from Bangladesh. Materials and Methods: Ethnobotanical information was collected from 16 kavirajes (traditional healers) in different study locations in Bangladesh. Plant samples were obtained from native habitats, botanical gardens, herbal markets and pharmaceutical companies. The samples were extracted using 70% methanol and were analysed using LC-DAD-MS and 1H-NMR. Results: Roots as well as leaves are commonly used for symptoms such as snake bites and sexual problems. Among the informants knowledge about toxicity or side effects is very limited and A. indica is often administered in very high doses. Replacement of A. indica with other medicinal plants such as Rauvolfia serpentina (L.) Benth. ex Kurz was common. A. indica samples contained a variety of aristolochic acid analogues such as aristolochic acid I, aristolochic acid II, cepharadione A and related compounds. Conclusions: AAN cases are likely to occur in Bangladesh and more awareness needs to be raised about the health risks associated with the use of A. indica and other species of Aristolochia as herbal medicines

The NASA-UC Eta-Earth Program: II. A Planet Orbiting HD 156668 with a Minimum Mass of Four Earth Masses

We report the discovery of HD 156668b, an extrasolar planet with a minimum mass of M_P sin i = 4.15 M_Earth. This planet was discovered through Keplerian modeling of precise radial velocities from Keck-HIRES and is the second super-Earth to emerge from the NASA-UC Eta-Earth Survey. The best-fit orbit is consistent with circular and has a period of P = 4.6455 d. The Doppler semi-amplitude of this planet, K = 1.89 m/s, is among the lowest ever detected, on par with the detection of GJ 581e using HARPS. A longer period (P ~ 2.3 yr), low-amplitude signal of unknown origin was also detected in the radial velocities and was filtered out of the data while fitting the short-period planet. Additional data are required to determine if the long-period signal is due to a second planet, stellar activity, or another source. Photometric observations using the Automated Photometric Telescopes at Fairborn Observatory show that HD 156668 (an old, quiet K3 dwarf) is photometrically constant over the radial velocity period to 0.1 mmag, supporting the existence of the planet. No transits were detected down to a photometric limit of ~3 mmag, ruling out transiting planets dominated by extremely bloated atmospheres, but not precluding a transiting solid/liquid planet with a modest atmosphere.Comment: This planet was announced at the 2010 AAS meeting in Wash. DC; 12 pages, 8 figures, 3 tables, submitted to Ap

Mechanisms in mutualisms: a chemically mediated thrips pollination strategy in common elder

The concept of flower-feeding thrips as pollinating insects in temperate regions is rarely considered as thrips are more frequently regarded to be destructive florivores feeding on pollen and surrounding plant tissue. Combining laboratory and field-based studies we examined interactions between Sambucus nigra (elderflower) and Thrips major within their native range to ascertain the role of thrips in the pollination of this species and to determine if floral chemicals mediated flower visits. If thrips provide a pollination service to S. nigra, then this will likely manifest in traits that attract the pollinating taxa at temporally critical points in floral development. T. major were highly abundant in inflorescences of S. nigra, entering flowers when stigmas were pollen-receptive and anthers were immature. When thrips were excluded from the inflorescences fruit-set failed. Linalool was the major component of the inflorescence headspace with peak abundance coinciding with the highest number of adult thrips visiting flowers. Thrips were absent in buds and their numbers declined again in senescing flowers correlating with the concentration of cyanogenic glycosides recorded in the floral tissue. Our data show that S. nigra floral chemistry mediates the behaviour of pollen-feeding thrips by attracting adults in high numbers to the flowers at pre-anthesis stage, while producing deterrent compounds prior to fruit development. Taking an integrative approach to studying thrips behaviour and floral biology we provide a new insight into the previously ambiguously defined pollination strategies of S. nigra and provide evidence that suggests that the relationship between T. major and S. nigra is mutualistic

Filling in the gaps of the papilionoid legume phylogeny: The enigmatic Amazonian genus Petaladenium is a new branch of the early-diverging Amburaneae clade

Recent deep-level phylogenies of the basal papilionoid legumes (Leguminosae, Papilionoideae) have resolved many clades, yet left the phylogenetic placement of several genera unassessed. The phylogenetically enigmatic Amazonian monospecific genus Petaladenium had been believed to be close to the genera of the Genistoid Ormosieae clade. In this paper we provide the first DNA phylogenetic study of Petaladenium and show it is not part of the large Genistoid clade, but is a new branch of the Amburaneae clade, one of the first-diverging lineages of the Papilionoideae phylogeny. This result is supported by the chemical observation that the quinolizidine alkaloids, a chemical synapomorphy of the Genistoids, are absent in Petaladenium. Parsimony and Bayesian phylogenetic analysis of nuclear ITS/5.8S and plastid matK and trnL intron agree with a new interpretation of morphology that Petaladenium is sister to Dussia, a genus comprising ~18 species of trees largely confined to rainforests in Central America and northern South America. Petaladenium, Dussia, and Myrospermum have papilionate flowers in a clade otherwise with radial floral symmetry, loss of petals or incompletely differentiated petals. Our phylogenetic analyses also revealed well-supported resolution within the three main lineages of the ADA clade (Angylocalyceae, Dipterygeae, and Amburaneae). We also discuss further molecular phylogenetic evidence for the undersampled Amazonian genera Aldina and Monopteryx, and the tropical African Amphimas, Cordyla, Leucomphalos, and Mildbraediodendron. © 2015 Elsevier Inc

Oral Abstracts 7: RA ClinicalO37. Long-Term Outcomes of Early RA Patients Initiated with Adalimumab Plus Methotrexate Compared with Methotrexate Alone Following a Targeted Treatment Approach

Background: This analysis assessed, on a group level, whether there is a long-term advantage for early RA patients treated with adalimumab (ADA) + MTX vs those initially treated with placebo (PBO) + MTX who either responded to therapy or added ADA following inadequate response (IR). Methods: OPTIMA was a 78- week, randomized, controlled trial of ADA + MTX vs PBO + MTX in MTX-naïve early (<1 year) RA patients. Therapy was adjusted at week 26: ADA + MTX-responders (R) who achieved DAS28 (CRP) <3.2 at weeks 22 and 26 (Period 1, P1) were re-randomized to withdraw or continue ADA and PBO + MTX-R continued randomized therapy for 52 weeks (P2); IR-patients received open-label (OL) ADA + MTX during P2. This post hoc analysis evaluated the proportion of patients at week 78 with DAS28 (CRP) <3.2, HAQ-DI <0.5, and/or ΔmTSS ≤0.5 by initial treatment. To account for patients who withdrew ADA during P2, an equivalent proportion of R was imputed from ADA + MTX-R patients. Results: At week 26, significantly more patients had low disease activity, normal function, and/or no radiographic progression with ADA + MTX vs PBO + MTX (Table 1). Differences in clinical and functional outcomes disappeared following additional treatment, when PBO + MTX-IR (n = 348/460) switched to OL ADA + MTX. Addition of OL ADA slowed radiographic progression, but more patients who received ADA + MTX from baseline had no radiographic progression at week 78 than patients who received initial PBO + MTX. Conclusions: Early RA patients treated with PBO + MTX achieved comparable long-term clinical and functional outcomes on a group level as those who began ADA + MTX, but only when therapy was optimized by the addition of ADA in PBO + MTX-IR. Still, ADA + MTX therapy conferred a radiographic benefit although the difference did not appear to translate to an additional functional benefit. Disclosures: P.E., AbbVie, Merck, Pfizer, UCB, Roche, BMS—Provided Expert Advice, Undertaken Trials, AbbVie—AbbVie sponsored the study, contributed to its design, and participated in the collection, analysis, and interpretation of the data, and in the writing, reviewing, and approval of the final version. R.F., AbbVie, Pfizer, Merck, Roche, UCB, Celgene, Amgen, AstraZeneca, BMS, Janssen, Lilly, Novartis—Research Grants, Consultation Fees. S.F., AbbVie—Employee, Stocks. A.K., AbbVie, Amgen, AstraZeneca, BMS, Celgene, Centocor-Janssen, Pfizer, Roche, UCB—Research Grants, Consultation Fees. H.K., AbbVie—Employee, Stocks. S.R., AbbVie—Employee, Stocks. J.S., AbbVie, Amgen, AstraZeneca, BMS, Celgene, Centocor-Janssen, GlaxoSmithKline, Lilly, Pfizer (Wyeth), MSD (Schering-Plough), Novo-Nordisk, Roche, Sandoz, UCB—Research Grants, Consultation Fees. R.V., AbbVie, BMS, GlaxoSmithKline, Human Genome Sciences, Merck, Pfizer, Roche, UCB Pharma—Consultation Fees, Research Support. Table 1.Week 78 clinical, functional, and radiographic outcomes in patients who received continued ADA + MTX vs those who continued PBO + MTX or added open-label ADA following an inadequate response ADA + MTX, n/N (%)a PBO + MTX, n/N (%)b Outcome Week 26 Week 52 Week 78 Week 26 Week 52 Week 78 DAS28 (CRP) <3.2 246/466 (53) 304/465 (65) 303/465 (65) 139/460 (30)*** 284/460 (62) 300/460 (65) HAQ-DI <0.5 211/466 (45) 220/466 (47) 224/466 (48) 150/460 (33)*** 203/460 (44) 208/460 (45) ΔmTSS ≤0.5 402/462 (87) 379/445 (86) 382/443 (86) 330/459 (72)*** 318/440 (72)*** 318/440 (72)*** DAS28 (CRP) <3.2 + ΔmTSS ≤0.5 216/462 (47) 260/443 (59) 266/443 (60) 112/459 (24)*** 196/440 (45) 211/440 (48)*** DAS28 (CRP) <3.2 + HAQ-DI <0.5 + ΔmTSS ≤0.5 146/462 (32) 168/443 (38) 174/443 (39) 82/459 (18)*** 120/440 (27)*** 135/440 (31)** aIncludes patients from the ADA Continuation (n = 105) and OL ADA Carry On (n = 259) arms, as well as the proportional equivalent number of responders from the ADA Withdrawal arm (n = 102). bIncludes patients from the MTX Continuation (n = 112) and Rescue ADA (n = 348) arms. Last observation carried forward: DAS28 (CRP) and HAQ-DI; Multiple imputations: ΔmTSS. ***P < 0.001 and **iP < 0.01, respectively, for differences between initial treatments from chi-squar

Characterisation of phenylethanoid glycosides by multiple‐stage mass spectrometry

Rationale: Although phenylethanoid glycosides (PhGs) occur widely in plants, their characterisation by liquid chromatography/mass spectrometry (LC/MS) is less well studied than other phenolic glycosides such as flavonoid glycosides. The multiple‐stage mass spectrometry (MSn) experiments required to improve the annotation of common verbascoside‐type PhGs are described here. Methods: Deprotonated, ammoniated and sodiated molecules of nine PhGs were subjected to low‐energy collision‐induced dissociation (CID) in a hybrid ion trap/orbitrap mass spectrometer. Most experiments were recorded at nominal mass using the linear ion trap analyser for wider applicability in the plant metabolomics community. Data interpretation was supported by high‐resolution orbitrap scanning of product ions. Comparative data was acquired on the same instrument by performing higher‐energy collisional dissociation (HCD) in the C‐trap. Results: Low energy CID‐MS2 of the deprotonated and ammoniated molecules generated diagnostic product ions from which the molecular masses of the phenolic acid and phenylethanoid moieties, respectively, could be determined. The sugar at C‐3' of the core glucose was preferentially lost from the sodiated molecule following CID‐MS2, while CID‐MSn produced a sodiated product ion from ring cleavage of the core glucose bearing the sugar at C‐6'. Evidence of a disaccharide substitution came from a sodiated disaccharide residue in CID‐MSn spectra. Conclusions: The consistency of PhG dissociation following low‐energy CID‐MSn of various ions is sufficient to enable annotation of verbascoside‐type PhGs in LC/MS analyses of crude plant extracts. This can be achieved on a low‐resolution instrument capable of MSn