Characterization of Prorocentrum elegans and Prorocentrum levis (Dinophyceae) from the southeastern Bay of Biscay by morphology and molecular phylogeny

Benthic Prorocentrum species can produce toxins that adversely affect animals and human health. They are known to co‐occur with other bloom‐forming, potentially toxic, benthic dinoflagellates of the genera Ostreopsis, Coolia, and Gambierdiscus. In this study, we report on the presence of P. elegans M.Faust and P. levis M.A.Faust, Kibler, Vandersea, P.A. Tester & Litaker from the southeastern Bay of Biscay. Sampling was carried out in the Summer‐Autumn 2010–2012 along the Atlantic coast of the Iberian Peninsula, but these two species were only found in the northeastern part of the Peninsula. Strains were isolated from macroalgae collected from rocky‐shore areas bordering accessible beaches. Morphological traits of isolated strains were analyzed by LM and SEM, whereas molecular analyses were performed using the LSU and internal transcribed spacer (ITS)1‐5.8S‐ITS2 regions of the rDNA. A bioassay with Artemia fransciscana and liquid chromatography–high‐resolution mass spectrometry analyses were used to check the toxicity of the species, whose results were negative. The strains mostly corresponded to their species original morphological characterization, which is supported by the phylogenetic analyses in the case of P. levis, whereas for P. elegans, this is the first known molecular characterization. This is also the second known report of P. elegans.     

Same-Day Diagnosis Based on Histology for Women Suspected of Breast Cancer: High Diagnostic Accuracy and Favorable Impact on the Patient

Background

Same-day diagnosis based on histology is increasingly being offered to patients suspected of breast cancer. We evaluated to which extent same-day diagnosis affected diagnostic accuracy and patients'' anxiety levels during the diagnostic phase.

Patients and methods

All 759 women referred for same-day evaluation of suspicious breast lesions between November 2011–March 2013 were included. Diagnostic accuracy was assessed by linking all patients to the national pathology database to identify diagnostic discrepancies, in which case slides were reviewed. Patients'' anxiety was measured in 127 patients by the State Trait and Anxiety Inventory on six moments during the diagnostic workup and changes over time (< = 1 week) were analyzed by mixed effect models.

Results

Core-needle biopsy was indicated in 374/759 patients (49.3%) and in 205/759 (27%) patients, invasive or in situ cancer was found. Final diagnosis on the same day was provided for 606/759 (79.8%) patients. Overall, 3/759 (0.4%) discordant findings were identified. Anxiety levels decreased significantly over time from 45.2 to 30.0 (P = <0.001). Anxiety levels decreased from 44.4 to 25.9 (P = <0.001) for patients with benign disease, and remained unchanged for patients diagnosed with malignancies (48.6 to 46.7, P = 0.933). Time trends in anxiety were not affected by other patient or disease characteristics like age, education level or (family) history of breast cancer.

Conclusion

Same-day histological diagnosis is feasible in the vast majority of patients, without impairing diagnostic accuracy. Patients'' anxiety rapidly decreased in patients with a benign diagnosis and remained constant in patients with malignancy.     

HDL Size is More Accurate than HDL Cholesterol to Predict Carotid Subclinical Atherosclerosis in Individuals Classified as Low Cardiovascular Risk

Background

Misclassification of patients as low cardiovascular risk (LCR) remains a major concern and challenges the efficacy of traditional risk markers. Due to its strong association with cholesterol acceptor capacity, high-density lipoprotein (HDL) size has been appointed as a potential risk marker. Hence, we investigate whether HDL size improves the predictive value of HDL-cholesterol in the identification of carotid atherosclerotic burden in individuals stratified to be at LCR.

Methods and Findings

284 individuals (40–75 years) classified as LCR by the current US guidelines were selected in a three-step procedure from primary care centers of the cities of Campinas and Americana, SP, Brazil. Apolipoprotein B-containing lipoproteins were precipitated by polyethylene glycol and HDL size was measured by dynamic light scattering (DLS) technique. Participants were classified in tertiles of HDL size (<7.57; 7.57–8.22; >8.22 nm). Carotid intima-media thickness (cIMT) <0.90 mm (80^th percentile) was determined by high resolution ultrasonography and multivariate ordinal regression models were used to assess the association between cIMT across HDL size and levels of lipid parameters. HDL-cholesterol was not associated with cIMT. In contrast, HDL size >8.22 nm was independently associated with low cIMT in either unadjusted and adjusted models for age, gender and Homeostasis Model Assessment 2 index for insulin sensitivity, ethnicity and body mass index (Odds ratio 0.23; 95% confidence interval 0.07–0.74, p = 0.013).

Conclusion

The mean HDL size estimated with DLS constitutes a better predictor for subclinical carotid atherosclerosis than the conventional measurements of plasma HDL-cholesterol in individuals classified as LCR.     

Co-Exposure with Fullerene May Strengthen Health Effects of Organic Industrial Chemicals

In vitro toxicological studies together with atomistic molecular dynamics simulations show that occupational co-exposure with C₆₀ fullerene may strengthen the health effects of organic industrial chemicals. The chemicals studied are acetophenone, benzaldehyde, benzyl alcohol, m-cresol, and toluene which can be used with fullerene as reagents or solvents in industrial processes. Potential co-exposure scenarios include a fullerene dust and organic chemical vapor, or a fullerene solution aerosolized in workplace air. Unfiltered and filtered mixtures of C₆₀ and organic chemicals represent different co-exposure scenarios in in vitro studies where acute cytotoxicity and immunotoxicity of C₆₀ and organic chemicals are tested together and alone by using human THP-1-derived macrophages. Statistically significant co-effects are observed for an unfiltered mixture of benzaldehyde and C₆₀ that is more cytotoxic than benzaldehyde alone, and for a filtered mixture of m-cresol and C₆₀ that is slightly less cytotoxic than m-cresol. Hydrophobicity of chemicals correlates with co-effects when secretion of pro-inflammatory cytokines IL-1β and TNF-α is considered. Complementary atomistic molecular dynamics simulations reveal that C₆₀ co-aggregates with all chemicals in aqueous environment. Stable aggregates have a fullerene-rich core and a chemical-rich surface layer, and while essentially all C₆₀ molecules aggregate together, a portion of organic molecules remains in water.     

Genetic Inactivation of European Sea Bass (Dicentrarchus labrax L.) Eggs Using UV-Irradiation: Observations and Perspectives

Androgenesis is a form of uniparental reproduction leading to progenies inheriting only the paternal set of chromosomes. It has been achieved with variable success in a number of freshwater species and can be attained by artificial fertilization of genetically inactivated eggs following exposure to gamma (γ), X-ray or UV irradiation (haploid androgenesis) and by restoration of diploidy by suppression of mitosis using a pressure or thermal shock. The conditions for the genetic inactivation of the maternal genome in the European sea bass (Dicentrarchus labrax L.) were explored using different combinations of UV irradiation levels and durations. UV treatments significantly affected embryo survival and generated a wide range of developmental abnormalities. Despite the wide range of UV doses tested (from 7.2 to 720 mJ.cm⁻²), only one dose (60 mJ.cm⁻².min⁻¹ with 1 min irradiation) resulted in a small percentage (14%) of haploid larvae at hatching in the initial trials as verified by flow cytometry. Microsatellite marker analyses of three further batches of larvae produced by using this UV treatment showed a majority of larvae with variable levels of paternal and maternal contributions and only one larva displaying pure paternal inheritance. The results are discussed also in the context of an assessment of the UV-absorbance characteristics of egg extracts in this species that revealed the presence of gadusol, a compound structurally related to mycosporine-like amino acids (MAAs) with known UV-screening properties.     

Effects of elevated carbon dioxide, ozone and water availability on spring wheat growth and yield

Spring wheat (Triticum aestivum L. cv. Dragon) was exposed to elevated carbon dioxide (CO₂), alone (1995) or in combination with two levels of increased ozone (O₃) (1994) or increased irrigation (1996) during three successive growing seasons as part of the EU ESPACE‐wheat programme and conducted in open‐top chambers (OTCs) and ambient air (AA) plots at Östad, 50 km north‐east of Göteborg, Sweden. Doubling the CO₂ concentration had a positive effect on grain yield in all 3 years (+21, +7 and +11%, respectively), although only statistically significant in 1994. That year was characterised by a warm and dry summer in comparison with 1995 and 1996, in which the summers were more humid and typical for south‐west Sweden. In 1994, the CO₂‐induced increase in grain yield was associated with an increase in the duration of the green leaf area, a positive effect on straw yield and on the number of ears per square metre and a negative effect (?13%) on grain protein concentration. Harvest index was unaffected by the elevated CO₂ concentration. The only statistically significant effect of elevated CO₂ in 1995 was a decrease in the grain protein concentration (?11% in both CO₂ concentrations), and in 1996 an increase (+21%) in the straw yield. In 1996 the soil water potential was less negative in elevated CO₂, which is likely to reflect a lower water consumption of these plants. Addition of extra O₃ significantly affected the grain yield (?6 and ?10%, respectively) and the 1 000‐grain weight negatively (?3 and ?6%). Statistically significant interactions between CO₂ and O₃ were obtained for the number of ears per unit area and for the 1 000‐grain weight. The 1 000‐grain weight was negatively affected by O₃ in low CO₂, but remained unaffected in the high CO₂ treatment. There was a significant decrease (?6%) in the grain protein concentration induced by elevated irrigation. The chambers, compared with AA plots, had a positive effect on plant development and on grain yield in all 3 years.     

Three levels of integrating ecology with the conservation of South American temperate forests: the initiative of the Institute of Ecological Research Chiloé, Chile

The diversity of native species assemblages and that of indigenous cultures that once characterized the temperate forests of southern Chile have experienced a process of homogenization ever since the Spanish conquest. Today this process continues to erode both biotic and cultural diversity. With the goal of linking ecological research with actions to conserve the biological and cultural richness of this region, we established the Institute of Ecological Research Chiloé. The Institute's philosophy and activities involve three approaches: (1) participation of professional ecologists in environmental education and decision making, through collaboration with the community at local, regional, and global scales. (2) programs of ecological education, which include planting indigenous trees in urban areas and creating a local botanical garden with representative Chilean forest species. (3) critical analyses of the narrow economic and utilitarian environmental ethics that currently prevail in Chile, and often in other Latin American nations, and examination of traditional or novel alternative ethics and perspectives that address multiple interrelations between biological and cultural dimensions.     

Buffer zones for forest reserves: opinions of land owners and conservation value of their forest around nature reserves in southern Sweden

Buffer zones around reserves are often suggested, but have rarely been evaluated. We examined their design for small forest reserves (5–225 ha), where buffer zones (200 m wide) would protect the reserves and reduce negative edge effects. The potential buffer zones could be partly protected, but remain as private land. Alternatively, the state may buy land outside reserves (to be included in reserves). To consider opinions of forest owners, we interviewed 33 private forest owners in potential buffer zones of reserves. The respondents were weakly positive to conservation, but disliked a state reserve on their land, or a buffer zone where 50% of the forest would be protected without compensation; however, with compensation the majority of them were neutral or positive to such a buffer zone. In a choice between buffer zone or reserve bought by the state, the great majority of the respondents chose the buffer zone (compensation paid, land still private). We found no relationships between opinions of respondents and conservation values of their forests (densities of old and dead trees), but older owners had forests of higher value. The forest in the potential buffer zones had higher conservation value than other non-protected forests in southern Sweden. Thus, buffer zones may be valuable, and we suggest they are created in cooperation with local residents where this is possible.     

Fibroblast-like cells from rat plantar skin and neurotrophin-transfected 3T3 fibroblasts influence neurite growth from rat sensory neurons in vitro

Our previous finding that skin-derived and muscle-derived molecules can be used to sort regenerating rat sciatic nerve axons evoked questions concerning neuron-target interactions at the level of single cells, which prompted the present study. The results show that dorsal root ganglion (DRG) neurons co-cultured with fibroblast-like skin-derived cells emit many neurites. These have a proximal linear segment and a distal network of beaded branches in direct relation to skin-derived cells. Electron microscopic examination of such co-cultures showed bundles of neurites at some distance from the target cells and single profiles closely apposed to subjacent cells. RNase protection assay revealed that cultivated skin-derived cells express nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4). In co-cultures of DRG neurons and 3T3 fibroblasts overexpressing either of the neurotrophins produced by skin-derived cells the picture varied. NT-3 transfected 3T3 fibroblasts gave a growth pattern similar to that seen with skin-derived cells. Neurons co-cultured with mock-transfected 3T3 fibroblasts were small and showed weak neurite growth. In co-cultures with a membrane insert between skin-derived cells or 3T3 fibroblasts and DRG neurons few neurons survived and neurite growth was very sparse. We conclude that skin-derived cells stimulate neurite growth from sensory neurons in vitro, that these cells produce NGF, BDNF, NT-3 and NT-4 and that 3T3 fibroblasts producing NT-3 mimic the effect of skin-derived cells on sensory neurons in co-culture. Finally the results suggest that cell surface molecules are important for neuritogenesis.     

Decreased Expression Of apM1 in Omental and Subcutaneous Adipose Tissue of Humans With Type 2 Diabetes

We have screened a subtracted cDNA library in order to identify differentially expressed genes in omental adipose tissue of human patients with Type 2 diabetes. One clone (#1738) showed a marked reduction in omental adipose tissue from patients with Type 2 diabetes. Sequencing and BLAST analysis revealed clone #1738 was the adipocyte-specific secreted protein gene apM1 (synonyms ACRP30, AdipoQ, GBP28). Consistent with the murine orthologue, apM1 mRNA was expressed in cultured human adipocytes and not in preadipocytes. Using RT-PCR we confirmed that apM1 mRNA levels were significantly reduced in omental adipose tissue of obese patients with Type 2 diabetes compared with lean and obese normoglycemic subjects. Although less pronounced, apM1 mRNA levels were reduced in subcutaneous adipose tissue of Type 2 diabetic patients. Whereas the biological function of apM1 is presently unknown, the tissue specific expression, structural similarities to TNFα and the dysregulated expression observed in obese Type 2 diabetic patients suggest that this factor may play a role in the pathogenesis of insulin resistance and Type 2 diabetes.