Percutaneous cryoablation of colorectal liver metastases: potentiated by two consecutive freeze-thaw cycles

Cryoablation may be beneficial for selected patients with liver tumours. Two freeze-thaw cycles at the same location have been recommended during treatment as this potentiate the effect of ablation in experimental studies. However, single freeze ablations are used by some as double freeze procedures are time-consuming and have been associated with increased risk of complications. Estimation of ice-ball volume is difficult using regularly used monitoring techniques. Magnetic resonance imaging, however, allows excellent and multiplanar visualisation of the frozen region during ablation. We comment on the effect of double freeze cycles in regard to ice-ball volume as estimated from magnetic resonance imaging during percutaneous cryoablation of colorectal liver metastases. The ice-ball volume at the end of the second freeze cycle was median 42% larger than the volume at the end of the first freeze. Double freeze cycles may thus facilitate tumour destruction.     

Molecular Phylogeny of Some European Heteronemertean (Nemertea) Species and the Monophyletic Status ofRiseriellus, Lineus,andMicrura

The 16S rRNA mitochondrial gene was used to reconstruct the relationships among 10 heteronemertean species (subclass Heteronemertea, phylum Nemertea);Lineus ruberandL. viridisare represented by more than one specimen to assess intraspecific variation in these enigmatic species, and the analysis includes in total 14 terminal taxa incorporating one palaeonemertean species (Tubulanus annulatus) for outgroup rooting. The aligned sequences were subjected to maximum parsimony, maximum-likelihood, and neighbor-joining analyses to estimate the phylogenetic relationship of the species. The results were concordant from all analyses and indicate that neitherLineusnorMicruraare monophyletic taxa, and that there is no support from a phylogenetic point of view to establish the monotypic genusRiseriellus.     

Linker insertion analysis of the FimH adhesin of type 1 fimbriae in an Escherichia coli fimH-null background

Abstract The gene encoding the Escherichia coli FimH adhesin of type 1 fimbriae has been subjected to linker insertion mutagenesis. Amino acid changes were introduced at a number of positions spanning the entire sequence in order to probe the structure-function relationship of the FimH protein. The effect of these mutations on the ability of bacteria to express a D-mannose binding phenotype was assessed in a fimH null mutant (MS4) constructed by allelic exchange in the E. coli K-12 strain PC31. Mutations mapping at amino acid residues 36, 58 and 279 of the mature FimH protein were shown to completely abolish binding to D-mannose receptors. Differences in the level of fimbriation were also observed as a result of some of the mutations in the fimH gene. These mutants may prove useful in dissecting receptor-ligand interactions by defining regions of the FimH protein that are important in erythrocyte binding.     

Effect of grazing by a neotropical copepod, Notodiaptomus, on a natural cyanobacterial assemblage and on toxic and non-toxic cyanobacterial strains

The neotropical, freshwater copepod Notodiaptomus iheringi decreasedthe growth of small colonies in cyanobacteria-dominated plankton,indicating that they can be efficiently ingested and that theymay represent an important food source for this copepod in reservoirsdominated by cyanobacteria. Cultured, colonial, toxic Microcystisaeruginosa was not affected, indicating that toxicity may causea decrease in its ingestion by N. iheringi.     

Purification and Characterization of High Molecular Weight Human Pituitary Prolactin

A large form of human prolactin (molecular weight 150 000–170 000) was purified from the residue remaining after extraction at neutral pH of homogenized frozen pituitaries. This purification involved extraction at pH 9.8, molecular sieve chromatography on Sepharose CL-6B, and hydrophobic interaction chromatography on pentyl-Sepharose 4B. The procedure was followed by radioimmunoassay. The large form of prolactin was prepared both from fresh and from long-term stored residues. In the latter case the final yield was considerably higher. By zone electrophoresis in agarose suspension the prolactin preparation was separated into four or five immunoactive components. In sedimentation equilibrium analysis in the ultracentrifuge, however, these isohormones showed heterogeneity, which was suggested to be caused by dissociation. Evaluation of data obtained from the bottom region of the cells gave molecular weight values of the components in the range of 160 000 – 180 000. One of the is hormones s further studied and exhibited bioactivity in the local crop-sac assay and showed an amino acid composition closely similar to that of the native monomer prolactin. The high molecular weight prolactin was partially dissociated by treatment with 50% ethylene glycol or 1 M propionic acid or 6 M guanidine hydrochloride. Molecular sieve chromatography in the presence of these dissociating agents, resolved the prolactin activity into three separate peaks. The most retarded fraction, which eluted in a position corresponding to that of native monomer prolactin was characterized by electrophoresis and amino acid analysis. The results were supporting evidence that the dissociation procedure gave a monomer which had a lower amide content than the native monomer. Furthermore, its specific immunoactivity was 2–3 times higher than the activity of the intact large form.     

SOMA: A Single Oligonucleotide Mutagenesis and Cloning Approach

Modern biology research requires simple techniques for efficient and restriction site-independent modification of genetic material. Classical cloning and mutagenesis strategies are limited by their dependency on restriction sites and the use of complementary primer pairs. Here, we describe the Single Oligonucleotide Mutagenesis and Cloning Approach (SOMA) that is independent of restriction sites and only requires a single mutagenic oligonucleotide to modify a plasmid. We demonstrate the broad application spectrum of SOMA with three examples. First, we present a novel plasmid that in a standardized and rapid fashion can be used as a template for SOMA to generate GFP-reporters. We successfully use such a reporter to assess the in vivo knock-down quality of morpholinos in Xenopus laevis embryos. In a second example, we show how to use a SOMA-based protocol for restriction-site independent cloning to generate chimeric proteins by domain swapping between the two human hRMD5a and hRMD5b isoforms. Last, we show that SOMA simplifies the generation of randomized single-site mutagenized gene libraries. As an example we random-mutagenize a single codon affecting the catalytic activity of the yeast Ssy5 endoprotease and identify a spectrum of tolerated and non-tolerated substitutions. Thus, SOMA represents a highly efficient alternative to classical cloning and mutagenesis strategies.     

Job strain and tobacco smoking: an individual-participant data meta-analysis of 166,130 adults in 15 European studies

Background

Tobacco smoking is a major contributor to the public health burden and healthcare costs worldwide, but the determinants of smoking behaviours are poorly understood. We conducted a large individual-participant meta-analysis to examine the extent to which work-related stress, operationalised as job strain, is associated with tobacco smoking in working adults.

Methodology and Principal Findings

We analysed cross-sectional data from 15 European studies comprising 166 130 participants. Longitudinal data from six studies were used. Job strain and smoking were self-reported. Smoking was harmonised into three categories never, ex- and current. We modelled the cross-sectional associations using logistic regression and the results pooled in random effects meta-analyses. Mixed effects logistic regression was used to examine longitudinal associations. Of the 166 130 participants, 17% reported job strain, 42% were never smokers, 33% ex-smokers and 25% current smokers. In the analyses of the cross-sectional data, current smokers had higher odds of job strain than never-smokers (age, sex and socioeconomic position-adjusted odds ratio: 1.11, 95% confidence interval: 1.03, 1.18). Current smokers with job strain smoked, on average, three cigarettes per week more than current smokers without job strain. In the analyses of longitudinal data (1 to 9 years of follow-up), there was no clear evidence for longitudinal associations between job strain and taking up or quitting smoking.

Conclusions

Our findings show that smokers are slightly more likely than non-smokers to report work-related stress. In addition, smokers who reported work stress smoked, on average, slightly more cigarettes than stress-free smokers.     

Ecological effects of lime treatment of acidified lakes and rivers in Sweden

Effects on the aquatic biota of lime (CaCO₃) application in acidified lakes and streams were studied in a number of waters. After treatment, lime-sensitive species of mosses (Sphagnum spp.) decreased, but species such as Potamogeton natans and Myriophyllum alterniflorum seemed to be favoured. A few years after liming species composition and diversity of phytoplankton, zooplankton and benthic insect larvae were almost identical to that found in oligotrophic and non-acid lakes. Molluscs and benthic crustaceans may have difficulties recolonizing. Reproduction of remaining species of fish was successful as soon as pH increased. High survival of larvae and fry can result in some extremely rich year classes with slow individual growth. In most cases restocking of depleted fish stocks was successful.     

Targeting of superantigens

The bacterial superantigen staphylococcal enterotoxin A (SEA) is an extremely potent activator of T lymphocytes when presented on MHC class II antigens. In order to induce T lymphocytes to reject a tumor, we substituted the specificity of SEA for MHC class II molecules with specificity for tumor cells by combining SEA with a MAb recognizing colon carcinomas. Chemical conjugates or recombinant fusion proteins of the MAb C215 and SEA retained excellent antigen binding properties whereas the binding to MHC class II was markedly reduced. The hybrid proteins directed SEA responsive T cells to tumors with specificity determined by the specificity of the MAb. Significant tumor cell killing was obtained at picomolar concentrations of the hybrid proteins and was the result of direct cell mediated by cytotoxicity as well as production of tumoricidal cytokines by T cells. Targeting of superantigens represents a novel approach to specific immunomodulation and deserves further study as a potential therapy for malignant disease.