Quantification of 1-hydroxypyrene, 1- and 2-hydroxynaphthalene, 3-hydroxybenzo[a]pyrene and 6-hydroxynitropyrene by HPLC-MS/MS in human urine as exposure biomarkers for environmental and occupational surveys

Context: Several urinary PAHs metabolites can be detected by HPLC-MS/MS for individual exposure assessment.

Objective: Quantitation of urinary metabolites of four PAHs, selected on the basis of their significance, with reduced costs and high sensitivity.

Materials and methods: HPLC-MS/MS was used and pure standards and isotope-labeled internal analogs of the analytes. Two hundred samples were tested after enzymatic hydrolysis.

Results: Accuracy was higher than 90% and variability lower than 19%; LODs permit to measure 1-hydroxypyrene, 1 and 2-hydroxynaphthalene in all subjects, 6-hydroxynitropyrene in the 65% and 3-hydroxybenzo[a]pyrene in the 70%.

Discussion and conclusion: The method is suitable both for occupational and for environmental studies. This is the first paper reporting urinary levels of 6-hydroxynitropyrene in European subjects, nonoccupationally exposed to nitro-PAHs.     

Mycorrhizal ecology on serpentine soils

Background: Serpentine ecosystems support different, often unique, plant communities; however, we know little about the soil organisms that associate with these ecosystems. Mycorrhizas, mutualistic symbioses between fungi and roots, are critical to nutrient cycling and energy exchange below ground.

Aims: We address three hypotheses: H1, diversity of mycorrhizal fungi in serpentine soils mirrors above-ground plant diversity; H2, the morphology of mycorrhizas and fungi on serpentine soils differs from that on non-serpentine; and H3, mycorrhizal fungal communities of the same or closely related hosts differ between serpentine and non-serpentine soils.

Methods: This review focuses on whether plant diversity on serpentine soils correlates with the below ground diversity of mycorrhizal fungi.

Results: Studies show that plants and fungi formed abundant ectomycorrhizal and arbuscular mycorrhizal symbioses on and off serpentine soils. No serpentine-endemic fungi were identified. Molecular analyses indicate distinct serpentine isolates for Cenococcum geophilum and for Acaulospora, suggesting adaptation to serpentine soils. While fungal sporocarp assemblages on serpentine sites resembled those off serpentine, fruiting of hypogeous fungi was greatly reduced.

Conclusions: Ectomycorrhizal fungal communities did not differ between soil types; however, arbuscular mycorrhizal communities differed in some cases but not others. The additive response to multiple factors, described as the serpentine syndrome, may explain part of the response by fungi.     

Plant sex and phenological stage affect interactions with rhizosphere nematode communities

Background: In dioecious plants, females and males associate differently with mycorrhizal fungi, but interactions with other rhizosphere organisms are less well studied.

Aims: We investigated the effect of plant sex on rhizosphere nematode communities associated with Corema album, a dioecious shrub occurring in coastal habitats.

Methods: Rhizosphere samples were collected from males and females in three populations (150 plants), during fruiting and flowering. Nematode communities were characterised and compared between plant sexes through statistical analyses of the abundance of trophic groups, plant parasitic nematode (PPN) genera and ecological indices.

Results: Free-living nematodes showed no statistically significant differences owing to plant sex. Conversely, PPN community composition was significantly different between plant sexes during fruiting but not flowering, suggesting that physiological requirements over the annual phenological cycle of the plant influence ecological interactions with the rhizosphere.

Of the 13 PPN genera identified, the ectoparasitic Criconema and Hemicriconemellawere more abundant in the rhizosphere of males during fruiting, whereas the endoparasitic Meloidogyne associated more frequently with females, suggesting that plant host suitability is related to PPN feeding strategy.

Conclusions: It appears that interactions of individuals of different sexes of C. album with the rhizosphere nematode community vary with phenological stage, especially for PPN.     

Effects of mycorrhizal colonization and elevated atmospheric carbon dioxide on carbon fixation and below-ground carbon partitioning in Plantago lanceolata

Plantago lanceolata with or without the mycorrhizal fungus Glomus mosseae were grown over a 100 d period under ambient (38050 mol mol^-1) and elevated (600150 mol mol^-1) atmospheric CO2 conditions. To achieve similar growth, non-mycorrhizal plants received phosphorus in solution whereas mycorrhizal plants were supplied with bonemeal. Measures of plant growth, photosynthesis and carbon input to the soil were obtained. Elevated CO2 stimulated plant growth to the same extent in mycorrhizal and non0mycorrhizal plants, but had no effect on the partitioning of carbon between shoots and roots or on shoot tissue phosphorus concentration. Mycorrhizal colonization was low, but unaffected by CO2 treatment. Net photosynthesis was stimulated both by mycorrhizal colonization and elevated CO2, and there was a more than additive effect of the two treatments on net photosynthesis. Colonization by mycorrhizal fungi inhibited acclimation, in terms of net carbon assimilation, or plants to elevated CO2. ¹³C natural abundance techniques were used to measure carbon input into the soil, although the results were not conclusive. Direct measurements of below-ground root biomass showed that elevated CO2 did stimulate carbon flow below-ground and this was higher in mycorrhizal than non-mycorrhizal plants. For the four treatment combinations, the observed relative differences in amount of below-ground carbon were compared with those expected from the differences in net photosynthesis. A considerable amount of the extra carbon fixed both as a result of mycorrhizal colonization and growth in elevated CO2 did not reveal itself as increased plant biomass. As there was no evidence for a substantial increase in soil organic matter, most of this extra carbon must have been respired by the mycorrhizal fungus and the roots or by the plants as dark-respiration. The need for detailed studies in this area is emphasized.     

The taxonomic status and range extension of Grimmia exquisita J.Muñoz (Grimmiaceae,Bryophyta)

Introduction. We found an unusual specimen collected in 1975 from Xinjiang Province, China which may represent Grimmia exquisita J.Muñoz. However, this species is known only from the type localities and its taxonomic status is still controversial.

Methods. A voucher of the Chinese specimen is deposited in IFP herbarium. This specimen was studied with the typical anatomical and morphological methods applied within Grimmia (Maier, 2010).

Key Results. The Chinese species represents G. exquisita. After examining the type specimens of G. exquisita, we confirmed that it should be recognised as a distinct species.

Conclusions. This study expands the distribution area of G. exquisita to East Asia. The Chinese specimen was discovered in a rather different habitat to its types and consequently, G. exquisita may be a more widely distributed species. Evaluation of the Grimmia specimens in many different herbaria is needed.     

Influence of soil pH on the soybean-endomycorrhiza symbiosis

Summary Soybean (Glycine max {L.} Merr.) cultivars were inoculated withGigaspora gigantea andGlomus mosseae to determine mycorrhizal: cultivar relationships as affected by soil pH. The specific cultivarfungal response was dependent on soil pH. Overall cultivar responses in unlimed soil (pH 5.1) were greater forG. gigantea thanG. mosseae. The Bossier —G. gigantea combination was particularly responsive in unlimed soil and showed an increase of 10% in shoot length, 35% in shoot dry weight. 75% in root dry weight, and 397% in nodule dry weight over uninoculated controls. Little cultivar response was observed withG. mosseae inoculation in unlimed soil. In limed soil (pH 6.2), the larger responses were obtained withG. mosseae inoculated plants, although inoculation with eitherG. mosseae orG. gigantea appeared effective. In general, nodulation was greater on mycorrhizal roots than on control roots.     

Reduced15N-nitrogen Transport Through Arbuscular Mycorrhizal Hyphae to Triticum aestivum L. Supplied with Ammonium vs. Nitrate Nutrition

This study compared the influence of or nutrition on the contribution of extraradical hyphae of the arbuscular mycorrhizal fungusGlomus mosseae(Nicol. and Gerd.) Gerd. and Trappe (BEG 107)to or uptake byTriticum aestivum L. ‘Hano’(summer wheat) with sufficient or insufficient N supply in semi-hydroponicculture. Roots and root-distant hyphae were spatially separatedin compartmentalized pots. Although -fed plants supplied with sufficient N had higher N concentrationsthan their -fed counterparts, this did not favourably affect colonization rates, hyphal lengthdensities or¹⁵N amounts transported via hyphae to the plants.Ammonium supply did not result in higher P or reduced carbohydrateconcentrations in the plants, so these factors could not explainthe reduced hyphal lengths. It was concluded that the effectof supply on hyphal length may be related to the reduced root growth and/or a direct effectof on hyphal growth. Plant N deficiency reduced the percentage root length colonized, hyphallength, total¹⁵N uptake by hyphae and dry weight of both - and -fed mycorrhizal plants. This was more obvious for -fed plants because plant biomass and hyphal lengthsof -fed plants were relatively low in mycorrhizal plants irrespective of the N concentrationsupplied. Copyright 2001 Annals of Botany Company Ammonium, arbuscular mycorrhiza, Glomus mosseae, nitrate, nitrogen uptake,¹⁵N studies, Triticum aestivum, wheat     

Feasibility of Phytoextraction to Clean Up Low-Level Uranium-Contaminated Soil

The potential to phytoextract uranium (U) from a sandy soil contaminated at low levels was tested in the greenhouse. Two soils were tested: a control soil (317 Bq ²³⁸U kg^-1) and the same soil washed with bicarbonate (69 Bq ²³⁸U kg^-1). Ryegrass (Lolium perenne cv. Melvina), Indian mustard (Brassica juncea cv. Vitasso), and Redroot Pigweed (Amarathus retroflexus) were used as test plants.

The annual removal of the soil activity with the biomass was less than 0.1%. The addition of citric acid (25 mmol kg^-1) 1 week before the harvest increased U uptake up to 500-fold. With a ryegrass and mustard yield of 15000 kg ha^-1 and 10000 kg ha^-1, respectively, up to 3.5% and 4.6% of the soil activity could annually be removed with the biomass.

With a desired activity reduction level of 1.5 and 5 for the bicarbonate washed and control soil, respectively, it would take 10 to 50 years to attain the release limit.

A linear relationship between the plant ²³⁸U concentration and the ²³⁸U concentration in the soil solution of the control, bicarbonate-washed, or citric acid-treated soil points to the importance of the soil solution activity concentration in determining U uptake and hence to the importance of solubilising agents to increase plant uptake.

However, citric acid addition resulted in a decreased dry weight production (all plants tested) and crop regrowth (in case of ryegrass).     

Flower colour variation in the montane plant Gentiana lutea L. (Gentianaceae) is unrelated to abiotic factors

Background: Flower colour variation among populations may result from the spatial variation of selective agents. The structure of phenotypic variation informs on the ecological processes related to this variation. Variation in floral traits is mainly attributed to variation in the pollinator fauna, while variation in vegetative traits is usually linked to abiotic factors or herbivores.

Aims: We investigated the geographical variation of flower colour (and correlated traits) in Gentiana lutea and the relationship with the variation in abiotic factors.

Methods: Phenotypic variation (flower colour, petal length, petal width, stalk length, leaf length, flower number, petal number and number of basal leaves) was assessed in 429 plants of 12 populations located at north-west Iberia. Additionally, we obtained data on the geographical coordinates, elevation, temperature, rainfall and radiation for each population.

Results: Populations mostly differed in flower colour, from orange to yellow from west to the east. Abiotic factors were unrelated to variation in either floral or vegetative traits.

Conclusions: Phenotypic variation among G. lutea populations does not result from adaptation to environmental factors. Other factors, such as historical events or selective pressure exerted by biotic interactions, might explain the flower colour variation in G. lutea along the Cantabrian Range.     

The northward shifting neophyte Tragopogon dubius is just as effective in forming mycorrhizal associations as the native T. pratensis

Background: As a consequence of climate warming, many organisms are shifting their range towards higher latitudes and altitudes. As not all do so at the same speed, this may disrupt biotic interaction. Release from natural enemies through range expansion can result in invasiveness, whereas loss of mutualists can reduce plant vigour and fitness. One of the most important groups of plant symbiotic mutualists is the arbuscular mycorrhizal fungi (AMF).

Aims: We used greenhouse experiments to test whether Tragopogon dubius, a species that has recently expanded its range northward and colonised the Netherlands, is able to associate with the same AMF as the native congener T. pratensis.

Methods: In soils collected from four locations in the new range of T. dubius we compared the density of infective AMF propagules associating with both plant species, as well as AMF colonisation of the roots. The AMF community structure in the roots of these species was also analysed using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE).

Results: Tragopogon dubius and T. pratensis did not differ in any of these characteristics.

Conclusions: We therefore conclude that the range-shifting T. dubius is as effective in the formation of mycorrhiza as the native congener.     

Two new species of Cololejeunea (Spruce) Schiffn. from Western Ghats,India

Introduction. During a recent floristic survey of liverworts from Western Ghats, India, two interesting liverworts of genus Cololejeunea were collected from forests in Mahabaleshwar in the state of Maharashtra.

Methods. After critical investigation, they have been described as new to science and both species, C. ghatensis G.Asthana & A.Srivastava sp. nov. and C. mehrana G.Asthana & A.Srivastava sp. nov. are illustrated and described here.

Key results. Cololejeunea ghatensis is an epiphyllous species growing on angiosperm leaves and has been collected near Chinaman Waterfall, Robbers Cave, Connaught Peak and Polo Ground area. Cololejeunea mehrana has been collected near Monkey point and the latter has an interesting habitat as it was found growing on moss leaves. Cololejeunea ghatensis is monoicous and C. mehrana is dioicous.     

C-Phycocyanin elicited antitumor efficacy via cell-cycle arrest,apoptosis induction,and invasion inhibition in esophageal squamous cell carcinoma

Objectives: Mounting evidence has demonstrated that C-Phycocyanin (C-PC) exhibits marked antitumor activity in a wide type of tumors, such as pancreas cancer, breast carcinoma, lung cancer, and colon cancer. The current study aimed to confirm the antitumor efficacy of C-PC in esophageal squamous cell carcinoma (ESCC).

Methods: The efficacy of C-PC was evaluated against the proliferation of ESCC cell lines EC9706 and EC1 by CCK-8 kit and in a mice model of ESCC EC9706. Cell cycle and apoptosis were investigated by flow cytometry, and cell invasion was determined via transwell chamber. Protein expression was examined by Western blots.

Results: We found that C-PC exhibited anti-proliferation ability in a time-dependent manner and a dose-dependent manner in ESCC EC9706 and EC1 cells. Besides, C-PC markedly arrested cell cycle in the G0/G1 phase, induced cell apoptosis and suppressed cell invasion ability in both EC9706 and EC1 cells (p?<?.01). Notably, C-PC evoked the elevations of Bax, PARP, and cleaved-caspase-3 protein, but reduced cyclin D1, CDK4, Bcl-2, MMP-2, and MMP-9 expression levels. Further investigation from in vivo experiment revealed that C-PC displayed significant antitumor efficacy in the xenografted EC9706 model.

Conclusions: Our data presented herein suggest C-PC exerts antitumor efficacy in ESCC.     

Gender-dependent differences of mitochondrial function and oxidative stress in rat skeletal muscle at rest and after exercise training

Objective: This study investigated gender-dependent differences of mitochondrial function and sensitivity to in vitro ROS exposure in rat skeletal muscle at rest and after exercise training.

Methods: Wistar rats underwent running training for 6 weeks. In vitro measurements of hydroxyl radical production, oxygen consumption (under basal and maximal respiration conditions) and ATP production were made on permeabilized fibers. Mitochondrial function was examined after exposure and non-exposure to an in vitro generator system of reactive oxygen species (ROS). Antioxidant enzyme activities and malondialdehyde (MDA) content were also determined.

Results: Compared with sedentary males, females showed a greater resistance of mitochondrial function (oxygen consumption and ATP production) to ROS exposure, and lower MDA content and antioxidant enzyme activities. The training protocol had more beneficial effects in males than females with regard to ROS production and oxidative stress. In contrast to male rats, the susceptibility of mitochondrial function to ROS exposure in trained females was unchanged.

Discussion: Exercise training improves mitochondrial function oxidative capacities in both male and female rats, but is more pronounced in males as a result of different mechanisms. The resistance of mitochondrial function to in vitro oxidative stress exposure and the antioxidant responses are gender- and training-dependent, and may be related to the protective effects of estrogen.     

Genetic variance of transforming growth factor β2 gene in conotruncal heart defects

Objective: The aim of this study was to evaluate the association between two haplotype-tag single nucleotide polymorphisms (SNPs) (rs6658835 and rs10495098) of TGF-β2 and conotruncal heart defects (CTDs).

Methods: Two polymorphisms of TGF-β2 gene were genotyped by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) from 259 CTDs patients and 310 control subjects.

Results: The association between SNP rs6658835 in TGF-β2 and CTDs has been found. The frequency of G allele in CTDs patients was significantly higher than that in control subjects (52.7% versus 40.3%, p?<?0.001, OR =1.649).

Conclusion: TGF-β2 gene polymorphisms may serve as a novel genetic marker for the risk of CTDs.     

Environmental factors influencing liana community diversity,structure and habitat associations in a tropical hill forest,Malaysia

Background: Although lianas are important in forest ecosystems, information on the environmental factors influencing their diversity, structure and habitat associations is scanty.

Aims: This study had the aim of determining variation in liana diversity, structure and habitat associations across different topographic habitats, and the environmental factors influencing them, in the Penang Hill Forest Reserve, Malaysia.

Methods: Liana diversity, structure and habitat associations were examined in 30 plots, 40 m × 40 m, randomly demarcated within three topographic habitats (flatland, slope and valley). The physico-chemical properties of the soils in the plots were determined and the altitude and slope of the plots recorded.

Results: Liana species richness, diversity and structure varied significantly among the topographic habitats. Liana species richness, diversity and abundance variations were related to soil moisture, phosphorus content, pH and altitude, whereas liana diameter and basal area were associated with soil magnesium and slope. A total of eight species had a positive association with the habitat and a further six showed a negative association. Four species had a positive association with the flatland habitat, but only three and two species, respectively, with slope and valley habitats. The slope habitat gave the highest number of species that showed a negative association.

Conclusions: Liana diversity and structure varied between habitats. Some liana species showed different associations with specific habitats. Environmental variables were identified that influence liana diversity, structure and habitat associations. The study provides important information about the factors influencing liana success, and these will be of value in liana conservation and management in tropical forests.     

Release kinetics of high-sensitivity cardiac troponins I and T and troponin T upstream open reading frame peptide (TnTuORF) in clinically induced acute myocardial infarction

Context: Troponin T upstream open reading frame peptide (TnTuORF) may be useful as a novel biomarker in acute cardiac syndromes.

Objective: The study examined the early release kinetics of TnTuORF.

Materials and methods: We analyzed the time course of the release of cardiac troponins I and T and TnTuORF in patients (n?=?31) with hypertrophic obstructive cardiomyopathy undergoing transcoronary ablation of septal hypertrophy (TASH).

Results: Fifteen minutes after TASH, the levels of both troponins increased significantly (cTnT median: 18?ng/L versus 27?ng/L; cTnI median: 15?ng/L versus 25?ng/L). TnTuORF showed no variation.

Discussion: We observed a significantly greater increase in cTnI compared with cTnT.

Conclusion: Our results demonstrate that troponin assays allow early detection of myocardial injury, whereas TnTuORF levels remain unchanged in this setting.     

Effect of lipid composition on incorporation of trastuzumab-PEG-lipid into nanoliposomes by post-insertion method: physicochemical and cellular characterization

Context: Anti-HER2 immunoliposomes are promising nanotechnology based systems for active targeting of breast tumors, which depends on the amount of incorporated antibody.

Objective/Aim: In this work, we investigated the possible effect of lipid composition on the incorporation of trastuzumab-PEG-PE micelles into nanoliposomes and on their subsequent specific cellular targeting.

Materials and methods: Trastuzumab (anti-HER2 monoclonal antibody) was monothiolated and conjugated to maleimide-PEG-PE micelles. Liposomes of different lipid compositions were prepared by the thin layer hydration. Trastuzumab-PEG-PE micelles were incorporated into the liposomes by the post-insertion method. The percentage of lipid mixing was determined based on fluorescence resonance energy transfer. Cellular binding and uptake of rhodamine-labeled immunoliposomes were studied in SKBR-3 (HER2⁺⁺⁺) and MCF-7 (HER2⁺) cells. Also, antitumor cell activity of the immunoliposomes was compared to free trastuzumab and the liposomes.

Results: The lipid mixing of trastuzumab-PEG-PE micelles depended on the liposome composition. The immunoliposomes containing DPPC, cholesterol and PEG-PE showed prominent lipid mixing. The lipid mixing was consistent with the cell binding results which showed an efficient and specific binding of the immunoliposomes to SKBR-3 cells. Antitumor cell activity of the immunoliposomes in SKBR-3, unlike MCF-7 cells, depended on the content of trastuzumab.

Discussion: Cholesterol and PEG-PE in the liposome composition are prerequisites for a successful lipid mixing due to their ability to facilitate fusion. The higher lipid mixing results in higher antibody incorporation and consequently higher targeted cell binding.

Conclusions: The lipid mixing depends on the liposome composition, which reflects targeted cell binding of the immunoliposomes.     

Association between polymorphisms in IL21 gene and risk for sepsis

Context: Sepsis is now the leading cause of death in the noncardiovascular intensive care unit (ICU).

Objective: To investigate whether polymorphisms in IL21 gene contribute to sepsis susceptibility.

Materials and methods: Three single-nucleotide polymorphisms of IL21 (rs907715, rs2055979, rs12508721) were genotyped by TaqMan assay in patients with sepsis and control subjects.

Results: Polymorphisms rs2055979 and rs12508721 in IL21 were more frequent in sepsis patients compared to general population. But allele frequency of rs907715 was not significantly different between sepsis patients and control subjects.

Conclusion: Polymorphisms in IL21 may be associated with sepsis risk.     

Experimental evidence from Cantabrian mountain heathlands suggests new recommendations for management of Vaccinium myrtillus L

Background: Management actions in mountain heathlands oriented to increasing the cover of Vaccinium myrtillus promote mowing of Calluna vulgaris to avoid competition. However, such action ignores the fact that plant–plant interactions range from competition to facilitation under different stress conditions.

Aims: To test whether the interactions between these two species are related to competition or facilitation, which would change the perspective for their management in mountain heathlands.

Methods: A total of 40 experimental plots located in a montane heathland (northern Spain) were selected. Calluna or Vaccinium plants were either cut at ground level or not cut, and plots were fertilized with nitrogen or not in a three-factorial design. Vegetative and reproductive shoot mass and length of the current year’s shoots were estimated in two consecutive years for both species. Herbivory on Vaccinium by browsing ungulates was compared in control plots and plots where Calluna had been cut 2 yrs ago.

Results: The two study species co-occurred in 72.4% of the sampled plots, although Calluna was more abundant. Vaccinium growth was greater in plots with Calluna than in plots where Calluna had been cut. Browsing on Vaccinium was also higher in plots were Calluna had been cut, and the effect of fertilization was only significant on the reproductive biomass of Calluna.

Conclusions: Contrary to initial proposals, our results suggest that Vaccinium does not always benefit from the cutting of Calluna in heathlands. Thus, the management of Cantabrian mountain heathlands should consider maintaining Calluna in order to facilitate the growth of Vaccinium.