Phylogeny and species delineation in the marine diatom Pseudo‐nitzschia (Bacillariophyta) using cox1, LSU,and ITS2 rRNA genes: A perspective in character evolution

Analyses of the mitochondrial cox1, the nuclear‐encoded large subunit (LSU), and the internal transcribed spacer 2 (ITS2) RNA coding region of Pseudo‐nitzschia revealed that the P. pseudodelicatissima complex can be phylogenetically grouped into three distinct clades (Groups I–III), while the P. delicatissima complex forms another distinct clade (Group IV) in both the LSU and ITS2 phylogenetic trees. It was elucidated that comprehensive taxon sampling (sampling of sequences), selection of appropriate target genes and outgroup, and alignment strategies influenced the phylogenetic accuracy. Based on the genetic divergence, ITS2 resulted in the most resolved trees, followed by cox1 and LSU. The morphological characters available for Pseudo‐nitzschia, although limited in number, were overall in agreement with the phylogenies when mapped onto the ITS2 tree. Information on the presence/absence of a central nodule, number of rows of poroids in each stria, and of sectors dividing the poroids mapped onto the ITS2 tree revealed the evolution of the recently diverged species. The morphologically based species complexes showed evolutionary relevance in agreement with molecular phylogeny inferred from ITS2 sequence–structure data. The data set of the hypervariable region of ITS2 improved the phylogenetic inference compared to the cox1 and LSU data sets. The taxonomic status of P. cuspidata and P. pseudodelicatissima requires further elucidation.     

Modeling cell entry into a micro-channel

Cell entry into a micro-channel has potential applications in cell sorting and cancer diagnostics. In this paper, we numerically model breast cancer cell entry into a constricted micro-channel. Our results indicate that the cell velocity decreases during entry and increases after entry, an observation in agreement with experiments. We found that the cell entry time depend strongly on the cortical stiffness and is minimum at some critical cortical elasticity. In addition, we found that for the same entry time, a stiff nucleus is displaced toward the cell front, whereas a viscous nucleus is displaced toward the rear. In comparison, the nucleus is less sensitive to the viscosity of the cytoplasm. These observations suggest that specific intra-cellular properties can be deduced non-invasively during cell entry, through the inspection of the nucleus using suitable illumination techniques, such as fluorescent labeling.     

Resveratrol derivative,trans‐3,5,4′‐trimethoxystilbene,exerts antiangiogenic and vascular‐disrupting effects in zebrafish through the downregulation of VEGFR2 and cell‐cycle modulation

Angiogenesis plays an important role in the development of neoplastic diseases such as cancer. Resveratrol and its derivatives exert antiangiogenic effects, but the mechanisms of their actions remain unclear. The aim of this study was to evaluate the antiangiogenic activity of resveratrol and its derivative trans‐3,5,4′‐trimethoxystilbene in vitro using human umbilical vein endothelial cells (HUVECs) and in vivo using transgenic zebrafish, and to clarify their mechanisms of action in zebrafish by gene expression analysis of the vascular endothelial growth factor (VEGF) receptor (VEGFR2/KDR) and cell‐cycle analysis. trans‐3,5,4′‐Trimethoxystilbene showed significantly more potent antiangiogenic activity than that of resveratrol in both assays. In zebrafish, trans‐3,5,4′‐trimethoxystilbene caused intersegmental vessel regression and downregulated VEGFR2 mRNA expression. Trans‐3,5,4′‐trimethoxystilbene also induced G2/M cell‐cycle arrest, most specifically in endothelial cells of zebrafish embryos. We propose that the antiangiogenic and vascular‐targeting activities of trans‐3,5,4′‐trimethoxystilbene result from the downregulation of VEGFR2 expression and cell‐cycle arrest at G2/M phase. J. Cell. Biochem. 109: 339–346, 2010. © 2009 Wiley‐Liss, Inc.     

Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation

The signal produced by fluorescence in situ hybridization (FISH) often is inconsistent among cells and sensitivity is low. Small DNA targets on the chromatin are difficult to detect. We report here an improved nick translation procedure for Texas red and Alexa Fluor 488 direct labeling of FISH probes. Brighter probes can be obtained by adding excess DNA polymerase I. Using such probes, a 30 kb yeast transgene, and the rp1, rp3 and zein multigene clusters were clearly detected.     

Mapping of a Magnaporthe grisea locus affecting rice (Oryza sativa) cultivar specificity

Magnaporthe grisea causes rice blast, the most important fungal disease of rice. The segregation of genes controlling virulence of M. grisea on rice was studied to establish the genetic basis of cultivar specificity in this host-parasite interaction. Full-sib progeny and parent isolates Guy11 and 2539 of M. grisea were inoculated onto rice (Oryza sativa) cultivar CO39 and five near-isogenic lines (NILs) of CO39. Each NIL contained a different single gene affecting resistance to specific isolates of M. grisea. No differential interactions between NILs and progeny or parents were observed; parents and progeny pathogenic on CO39 were pathogenic on all five NILs. Segregation ratios of 101 full-sib progeny, 117 progeny from full-sib parents, and 109 backcross progeny, indicated a common single gene affecting pathogenicity on CO39 and the five NILs. A subset of the above 327 isolates (43 fullsib progeny, 37 progeny from full-sib parents, and 32 backcross progeny) were inoculated onto rice cultivar 51583; all were pathogenic, indicating that cultivar specificity to CO39 was segregating in this population of isolates. The locus controlling cultivar specificity, named avrCO39, was mapped to chromosome 1 using a subset of the progeny previously used to construct an RFLP map of M. grisea. The closest reported RFLP markers were 11.8 (estimated 260 kb) and 17.2 cM (estimated 380 kb) away and provide starting points on either side of the locus for a chromosome walk to clone the locus.     

Effects of heat acclimatisation on work tolerance and thermoregulation in trained tropical natives

This study aimed to investigate the effects of heat acclimatisation on thermoregulatory responses and work tolerance in trained individuals residing in the tropics. Eighteen male trained soldiers, who are native to a warm and humid climate, performed a total of four heat stress tests donning the Skeletal Battle Order (SBO, 20.5 kg) and Full Battle Order (FBO, 24.7 kg) before (PRE) and after (POST) a 10-day heat acclimatisation programme. The trials were conducted in an environmental chamber (dry bulb temperature: 32 °C, relative humidity: 70%, solar radiation: 400 W/m²). Excluding the data sets of which participants fully completed the heat stress tests (210 min) before and after heat acclimatisation, work tolerance was improved from 173±30 to 201±18 min (∼21%, p<0.05, n=9) following heat acclimatisation. Following heat acclimatisation, chest skin temperature during exercise was lowered in SBO (PRE=36.7±0.3 vs. POST=36.5±0.3 °C, p<0.01) and FBO (PRE=36.8±0.4 vs. POST=36.6±0.3 °C, p<0.01). Ratings of perceived exertion were decreased with SBO and FBO (PRE=11±2; POST=10±2; p<0.05) after heat acclimatisation. Heat acclimatisation had no effects on baseline body core temperature, heart rate and sweat rate across trials (p>0.05). A heat acclimatisation programme improves work tolerance with minimal effects on thermoregulation in trained tropical natives.     

Increased activation of blood neutrophils after cigarette smoking in young individuals susceptible to COPD

Background

Cigarette smoking is the most important risk factor for Chronic Obstructive Pulmonary Disease (COPD). Only a subgroup of smokers develops COPD and it is unclear why these individuals are more susceptible to the detrimental effects of cigarette smoking. The risk to develop COPD is known to be higher in individuals with familial aggregation of COPD. This study aimed to investigate if acute systemic and local immune responses to cigarette smoke differentiate between individuals susceptible or non-susceptible to develop COPD, both at young (18-40 years) and old (40-75 years) age.

Methods

All participants smoked three cigarettes in one hour. Changes in inflammatory markers in peripheral blood (at 0 and 3 hours) and in bronchial biopsies (at 0 and 24 hours) were investigated. Acute effects of smoking were analyzed within and between susceptible and non-susceptible individuals, and by multiple regression analysis.

Results

Young susceptible individuals showed significantly higher increases in the expression of FcγRII (CD32) in its active forms (A17 and A27) on neutrophils after smoking (p = 0.016 and 0.028 respectively), independently of age, smoking status and expression of the respective markers at baseline. Smoking had no significant effect on mediators in blood or inflammatory cell counts in bronchial biopsies. In the old group, acute effects of smoking were comparable between healthy controls and COPD patients.

Conclusions

We show for the first time that COPD susceptibility at young age associates with an increased systemic innate immune response to cigarette smoking. This suggests a role of systemic inflammation in the early induction phase of COPD.

Trial registration

Clinicaltrials.gov: {"type":"clinical-trial","attrs":{"text":"NCT00807469","term_id":"NCT00807469"}}NCT00807469

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-014-0121-2) contains supplementary material, which is available to authorized users.