Impacts of environmental stress on resistance and resilience of algal‐associated bacterial communities

Algal‐associated bacteria are fundamental to the ecological success of marine green macroalgae such as Caulerpa. The resistance and resilience of algal‐associated microbiota to environmental stress can promote algal health and genetic adaptation to changing environments. The composition of bacterial communities has been shown to be unique to algal morphological niches. Therefore, the level of response to various environmental perturbations may in fact be different for each niche‐specific community. Factorial in situ experiments were set up to investigate the effect of nutrient enrichment and temperature stress on the bacterial communities associated with Caulerpa cylindracea. Bacteria were characterized using the 16S rRNA gene, and the community compositions were compared between different parts of the algal thallus (endo‐, epi‐, and rhizomicrobiome). Resistance and resilience were calculated to further understand the changes of microbial composition in response to perturbations. The results of this study provide evidence that nutrient enrichment has a significant influence on the taxonomic and functional structure of the epimicrobiota, with a low community resistance index observed for both. Temperature and nutrient stress had a significant effect on the rhizomicrobiota taxonomic composition, exhibiting the lowest overall resistance to change. The functional performance of the rhizomicrobiota had low resilience to the combination of stressors, indicating potential additive effects. Interestingly, the endomicrobiota had the highest overall resistance, yet the lowest overall resilience to environmental stress. This further contributes to our understanding of algal microbiome dynamics in response to environmental changes.     

Somatic embryogenic tissue establishment from mature Pinus nigra Arn. ssp. salzmannii embryos

Summary Somatic embryogenesis from mature zygotic embryos of salgare?o pine (Pinus nigra Arn. ssp. salzmannii) was induced after 2 wk of culture in L₁ medium [Murashige and Skoog mineral solution with 10.7 μM naphthaleneacetic acid (NAA) and 8.8 μM 6-benzyladenine (BA)] or L₂ medium [Gupta and Durzan mineral solution (DCR)] supplemented with the phytohormone combination as above. Four different combinations of growth regulators, NAA (2.6–10.7 μM) and BA (6.6–8.8 μM), were tried for subsequent passage. The best percentage of embryo induction and manifestation was obtained on DCR medium with 2.6 μM NAA and 6.6–8.8 μM BA. Transfer of isolated somatic embryos from the basal media to L₃ medium (Quoirin and Le Poivre modified mineral solution without hormones and supplemented with 0.3% activated charcoal), facilitated random embryo maturation and some development into plantlets.     

Influence of the sex of flowers on androgenesis in Aesculus hippocastanum L. anther culture

Summary Flowers of Aesculus hippocastanum L. are bisexual and zygomorphic, and are positioned on a 20–30 cm long inflorescence. Those located in the basal part of the panicle are female and fertile (segment A), flowers in the middle are bisexual (segment B), and those on top of the panicle are male (segment C). Androgenesis was achieved in anther culture which originated from three types of flowers cultured on modified Murashige and Skoog medium containing 2,4-dichlorophenoxyacetic acid (4.5 μM) and kinetin (4.6 μM). Differences in viability of uninuclear microspores were found between female (90.0%) and other flowers (bisexual 61.1%; male 72.7%.). Both the percentage of embryogenic anthers and the number of androgenic embryos formed per inflorescence differed according to the segment of origin. The highest embryogenic response was obtained in segment A (47.3%) and the lowest in segment C (24.1%). A significant difference was found between the number of androgenic embryos formed per inflorescence in segments A (921.0) and C (286.7). The highest germination percentage (21.3%) and plantelet formation (41.0%) were obtained on woody plant WPM liquid medium supplemented with 1% activated charcoal. Acclimation and regeneration were best from plantelets originating from female flowers (62.5%). Plantlets originating from bisexual and male flowers have much poorer survival (29.3 and 22.2%, respectively).     

NanoLC-FT-ICR MS improves proteome coverage attainable for approximately 3000 laser-microdissected breast carcinoma cells

Proteomics assays hold great promise for unraveling molecular events that underlie human diseases. Effective analysis of clinical samples is essential, but this task is considerably complicated by tissue heterogeneity. Laser capture microdissection (LCM) can be used to selectively isolate target cells from their native tissue environment. However, the small number of cells that is typically procured by LCM severely limits proteome coverage and biomarker discovery potential achievable by conventional proteomics platforms. Herein, we describe the use of nanoLC-FT-ICR MS for analyzing protein digests of 3000 LCM-derived tumor cells from breast carcinoma tissue, corresponding to 300 ng of total protein. A total of 2282 peptides were identified by matching LC-MS data to accurate mass and time (AMT) tag databases that were previously established for human breast (cancer) cell lines. One thousand and three unique proteins were confidently identified with two or more peptides. Based on gene ontology categorization, identified proteins appear to cover a wide variety of biological functions and cellular compartments. This work demonstrates that a substantial number of proteins can be detected and identified from limited number of cells using the AMT tag approach, and opens doors for high-throughput in-depth proteomics analysis of clinical samples.     

Improving the Electrokinetic Remediation of Nickel-, Cadmium-, and Lead-contaminated Sediment

In this work, the simultaneous application of bipolar electrodes and a longer cathodic compartment length without agents addition was used as an enhanced electrokinetic treatment of Ni-, Cd-, and Pb-contaminated sediment. Conventional and enhanced treatments were investigated and their efficacy assessed by comparing the residual metals content in the sediment with values given by Dutch standards. After conventional treatment removals of 20%, 19%, and 31% for Ni, Cd, and Pb were achieved, whereas the enhanced treatment achieved removal efficacies of 36% for Ni, 42% for Cd, and 43% for Pb. Results indicate that the simultaneous application of the techniques mentioned is effective for removing Ni and Pb whereas Cd was not removed to the desired level. Here we show that the addition of chemicals is not necessary to increase the efficacies of electrokinetic sediment treatments, so the simultaneous application of the techniques mentioned can be described as environmentally friendly.     

Association of the FAM46A Gene VNTRs and BAG6 rs3117582 SNP with Non Small Cell Lung Cancer (NSCLC) in Croatian and Norwegian Populations

We analyzed for associations between a variable number of tandem repeat (VNTR) polymorphism in the Family with sequence similarity 46, member A (FAM46A) gene and a single nucleotide polymorphism (rs3117582) in the BCL2-Associated Athanogene 6 (BAG6) with non small cell lung cancer in Croatian and Norwegian subjects. A total of 503 (262 Croatian and 241Norwegian) non small cell lung cancer patients and 897 controls (568 Croatian and 329 Norwegian) were analyzed. We found that the frequency of allele b (three VNTR repeats) of FAM46A gene was significantly increased in the patients compared to the healthy controls in the Croatian and the combined Croatian and Norwegian subjects. Genotype frequencies of cd (four and five VNTR repeats) and cc (four VNTR repeats homozygote) of the FAM46A gene were significantly decreased in the patients compared to the healthy controls in the Croatian and Norwegian subjects, respectively. Logistic regression analyses revealed FAM46A genotype cc to be an independent predictive factor for non small cell lung cancer risk in the Norwegian subjects after adjustment for age, gender and smoking status. This is the first study to suggest an association between the FAM46A gene VNTR polymorphisms and non small cell lung cancer. We found also that BAG6 rs3117582 SNP was associated with non small cell lung cancer in the Norwegian subjects and the combined Croatian-Norwegian subjects corroborating the earlier finding that BAG6 rs3117582 SNP was associated with lung cancer in Europeans. Logistic regression analyses revealed that genotypes and alleles of BAG6 were independent predictive factor for non small cell lung cancer risk in the Norwegian and combined Croatian-Norwegian subjects, after adjustment for age and gender.     

Biogeographical patterns of algal communities in the Mediterranean Sea: Cystoseira crinita‐dominated assemblages as a case study

Aim The aim of this study was to describe the composition, community structure and biogeographical variation of subtidal algal assemblages dominated by the brown alga Cystoseira crinita across the Mediterranean Sea. Location The Mediterranean coast, from Spain (1°25′ E) to Turkey (30°26′ E). Methods Data on the species composition and structure of assemblages dominated by the species C. crinita were collected from 101 sites in nine regions across the Mediterranean Sea. Multivariate and univariate statistical tools were used to investigate patterns of variation in the composition of the assemblages among sites and regions, and to compare these with previously defined biogeographical regions. Linear regressions of species richness versus longitude and versus latitude were also carried out to test previously formulated hypotheses of biodiversity gradients in the Mediterranean Sea. Results The main features characterizing C. crinita‐dominated assemblages across the Mediterranean included a similar total cover of species, a similar cover of C. crinita, and consistency in the presence of the epiphyte Haliptilon virgatum. Biogeographical variation was detected as shifts in relative abundances of species among regions, partly coinciding with previously described biogeographical sectors. A significant positive correlation was found between species richness and latitude, while no significant correlation was detected between species richness and longitude. Main conclusions The patterns of variation in community structure detected among the studied regions reflected their geographical positions quite well. However, latitude seemed to contribute more to the explanation of biological patterns of diversity than did geographical distances or boundaries, which classically have been used to delimit biogeographical sectors. Moreover, the positive correlation between species richness and latitude reinforced the idea that latitude, and possibly temperature as a related environmental factor, plays a primary role in structuring biogeographical patterns in the Mediterranean Sea. The lack of correlation between species richness and longitude contradicts the notion that there is a decrease in species richness from west to east in the Mediterranean, following the direction of species colonization from the Atlantic.     

Screening with mammography organized by family physicians teams: what have we learnt?

The mammography, recommended as standard method for screening on breast cancer, can reveal suspicious lesions early enough to anable cancer elimination in entirely. Experience with women of the target population, 50-69 years old, included in the mass screening programs, show the reduction in the specific mortality by 30%. One of the main problem in organizing the preventive programs is how to increase responsiveness of subjects to screening. In the study, based on the large sample of over 1000 of subjects and 20 family medicine practices, included in the investigation, we showed that it is possible, by a pro-active involvement of family physicians teams and intensive educational and motivational activities, to achieve high level of over 80% of responsiveness to mammography screening. Analysis of the reasons of nonresponsiveness can contribute to better understanding of the mental processes included in a self-decision making. This, as the final aim, can help family physicians in their efforts to overcome many hidden barriers which obstruct their patients to accept the mammography screening.     

Expression of Bcl-2 and Bax proteins in thyroid glands of rats in experimental thyroiditis

In the recent years, iodine was associated to the development of apoptosis in thyroid diseases. The aim of the present study is to determine the expression of pro-apoptotic and anti-apoptotic proteins, Bax and Bcl-2, in a Wistar rat experimental model of thyroiditis induced by administration of different doses of potassium iodide. Immunohistochemical staining was done with chromogen diaminobenzidine on avidin-biotin peroxidase using the Animal Research Kit (ARK), stained with antibodies to Bcl-2 and Bax proteins. The intensity and distribution of positive staining were evaluated by light microscopy on a scale of 0 to 4. Bax protein was expressed in the area of regenerating follicular cells in high percent in potassium iodide treated rats, but was not expressed in thyrocytes from control rats. Bcl-2 expression was constantly observed in thyrocytes of the control group and in the mantle-zone of lymphoid follicular infiltrates. Our results show that Bax expression is significantly higher in the Wistar rat experimental model of thyroiditis than in the control group. These data suggest that the increased expression of Bax may contribute to the role of apoptosis in the pathogenesis of experimental thyroiditis.     

Discovery and SAR study of novel dihydroquinoline containing glucocorticoid receptor ligands

We report the discovery of a novel class of glucocorticoid receptor (GR) ligands based on 1,2-dihydroquinoline molecular scaffold. The compounds exhibit good GR binding affinity and selectivity profile against other nuclear hormone receptors.