Determination of element distribution between the symplasm and apoplasm of cucumber plant parts by total reflection X-ray fluorescence spectrometry

The distribution of Cd, Ni, Pb and Fe between the symplasm (cytoplasm) and apoplasm (cell wall) of cucumber roots and leaves was determined by total reflection X-ray fluorescence spectrometry following a special sample preparation procedure. The plants were grown in modified Hoagland nutrient solution containing Fe in chemical form of Fe(III)-citrate or Fe(III)-EDTA, as well as the heavy metal contaminants, each in concentration of 10 microM. In the roots the larger part of Pb was found in the apoplasm, while Ni and Cd were mostly in the symplasm. In the leaves however, about 50-60% of the Pb content and practically the total amount of Cd were detected in the symplasm. About 30-40% of the translocated Ni remained in the apoplasm of the leaves. The Cd-, Ni- and Pb-treatments resulted in higher total concentration of Fe in the roots, however, the relative amount of Fe in the symplasm decreased in all cases. In the leaves of the control plants the larger part (60-80%) of Fe occurs in the symplasm. Due to the heavy metal effects, the relative amount of Fe in the symplasm decreased except in the Pb-contaminated plants, where in the presence of Fe(III)-EDTA, the Pb treatment resulted in a moderate increment of Fe concentration in the symplasm.     

MacroH2A histone variants limit chromatin plasticity through two distinct mechanisms

MacroH2A histone variants suppress tumor progression and act as epigenetic barriers to induced pluripotency. How they impart their influence on chromatin plasticity is not well understood. Here, we analyze how the different domains of macroH2A proteins contribute to chromatin structure and dynamics. By solving the crystal structure of the macrodomain of human macroH2A2 at 1.7 Å, we find that its putative binding pocket exhibits marked structural differences compared with the macroH2A1.1 isoform, rendering macroH2A2 unable to bind ADP‐ribose. Quantitative binding assays show that this specificity is conserved among vertebrate macroH2A isoforms. We further find that macroH2A histones reduce the transient, PARP1‐dependent chromatin relaxation that occurs in living cells upon DNA damage through two distinct mechanisms. First, macroH2A1.1 mediates an isoform‐specific effect through its ability to suppress PARP1 activity. Second, the unstructured linker region exerts an additional repressive effect that is common to all macroH2A proteins. In the absence of DNA damage, the macroH2A linker is also sufficient for rescuing heterochromatin architecture in cells deficient for macroH2A.     

Pyoverdine-mediated regulation of FpvA synthesis in Pseudomonas aeruginosa: involvement of a probable extracytoplasmic-function sigma factor,FpvI

A search of the pvd pyoverdine biosynthesis locus of Pseudomonas aeruginosa identified an open reading frame, PA2387, whose product exhibited a sequence similar to those of a number of so-called extracytoplasmic- function sigma factors responsible for siderophore-dependent expression of iron-siderophore receptors in Escherichia coli and Pseudomonas putida. Deletion of this gene, dubbed fpvI, compromised pyoverdine-dependent FpvA ferric pyoverdine receptor production and fpvA gene expression, while the cloned gene stimulated fpvA expression. A Fur-binding site was identified immediately upstream of fpvI, consistent with the observed iron-regulated expression of fpvI and fpvA.     

Phase II Open Label Study of Valproic Acid in Spinal Muscular Atrophy

Preliminary in vitro and in vivo studies with valproic acid (VPA) in cell lines and patients with spinal muscular atrophy (SMA) demonstrate increased expression of SMN, supporting the possibility of therapeutic benefit. We performed an open label trial of VPA in 42 subjects with SMA to assess safety and explore potential outcome measures to help guide design of future controlled clinical trials. Subjects included 2 SMA type I ages 2–3 years, 29 SMA type II ages 2–14 years and 11 type III ages 2–31 years, recruited from a natural history study. VPA was well-tolerated and without evident hepatotoxicity. Carnitine depletion was frequent and temporally associated with increased weakness in two subjects. Exploratory outcome measures included assessment of gross motor function via the modified Hammersmith Functional Motor Scale (MHFMS), electrophysiologic measures of innervation including maximum ulnar compound muscle action potential (CMAP) amplitudes and motor unit number estimation (MUNE), body composition and bone density via dual-energy X-ray absorptiometry (DEXA), and quantitative blood SMN mRNA levels. Clear decline in motor function occurred in several subjects in association with weight gain; mean fat mass increased without a corresponding increase in lean mass. We observed an increased mean score on the MHFMS scale in 27 subjects with SMA type II (p≤0.001); however, significant improvement was almost entirely restricted to participants <5 years of age. Full length SMN levels were unchanged and Δ7SMN levels were significantly reduced for 2 of 3 treatment visits. In contrast, bone mineral density (p≤0.0036) and maximum ulnar CMAP scores (p≤0.0001) increased significantly.

Conclusions

While VPA appears safe and well-tolerated in this initial pilot trial, these data suggest that weight gain and carnitine depletion are likely to be significant confounding factors in clinical trials. This study highlights potential strengths and limitations of various candidate outcome measures and underscores the need for additional controlled clinical trials with VPA targeting more restricted cohorts of subjects.

Trial Registration

ClinicalTrials.gov http://clinicaltrials.gov/ct2/show/NCT00374075?term=NCT00374075&rank=1     

Social Influence on Early Foraging of Domestic Chicks (Gallus gallus) in a Near-to-Nature Procedure

The aim of this study was to investigate the social influence of single chicks with different foraging experience on the behaviour of their group members during the first days after hatching. In contrast to the duplicate cage procedure normally used in social learning studies, four naïve group members could freely interact with a tutor or a control chick in an enriched 20-m² test arena. This arena contained two types of food caches with differently coloured food. The tutor was 3 d older than its group members and was trained to feed from one of the two food cache types, whereas the control chick was not familiar with the characteristics of the test arena and was either of the same age or 3 d older than its group members. During the first 9 d after hatching, the foraging activity of 32 chick groups was tested in 15 trials per group. Groups with a 3 d older but inexperienced control chick had the least foraging success whilst groups with a naïve control chick of the same age compared more favourably with the groups with experienced tutors than with groups that included older control chicks. In a second experiment, a tutor compartment was introduced into the test arena in order to compare a situation of unrestricted contact possibilities with the duplicate cage procedure in which tutor and bystanders are separated by a Perspex divider. The tutor compartment was opened for some test groups, allowing free interaction between tutor and naïve chicks. Those chicks which could only watch the tutor through the Perspex developed a preference for the tutor’s food type and discovered this food type as fast as the chicks of groups with unrestricted interactions. However, a relatively large proportion of groups (3 out of 8) did not develop successful foraging behaviour at all, whereas the successful groups showed longer feeding latencies during the subsequent trials than groups which had unrestricted contact with their tutor. This indicates an inhibiting effect of the duplicate cage procedure. It is concluded that when testing chicks of a few days of age, a distinction should be made between (i) the social influence on food particle preferences and (ii) the social influence on learning where to find food. This is important for generalizations taken from duplicate cage procedures without any environmental enrichment, as this restrictive setting only allows detection of food particle preferences.     

Erlotinib in the treatment of non-small cell lung cancer

Because erlotinib, an epidermal growth factor receptor tyrosine kinase (EGFRTK) inhibitor, has been shown to be effective and well-tolerated as a second/third-line treatment in the therapy of advanced non-small cell lung cancer (NSCLC), this agent has been recently approved for human NSCLC therapy in the European Union. Although additive and synergistic effects of erlotinib and conventional chemotherapy were demonstrated in the combined regime preclinically, this has yet to be approved in the clinic. Since erlotinib and cytotoxic drugs have different biological targets, they have distinct side effects as well: erlotinib has no toxic effect on the bone marrow, but can cause diarrhea and rash, the latter being thought to be an indicator of the therapeutic efficacy. Several ongoing clinical trials are investigating the potential role of erlotinib in different settings in human NSCLC. This review intends to integrate our current knowledge on the erlotinib treatment in NSCLC.     

Synthesis and stereochemical investigations of novel nitrogen-containing 13alpha-estrone derivatives

Novel tetrahydroquinoline 11 and N-aryl d-homo derivatives 12 in the 13alpha-estrone series were synthesized effectively, starting from the secoaldehyde 8 and mono- or disubstituted anilines 9. The chemoselectivity of the cyclization reactions depended upon the nature of the substituents in the anilines. All transformations proceeded in a highly stereoselective manner, yielding only one diastereomer. Condensed 11 and d-homo derivatives 12 both have the usual ring C chair conformation in the solid state.     

Changes of serum carotenoids in patients with esophageal, gastric, hepatocellular, pancreatic and colorectal cancer

The serum levels of carotenoids (vitamin A, lutein, zeaxanthin, alfa- and beta cryptoxanthin, alfa- and beta-carotene) were measured in healthy persons (n=40) and in 98 patients with different malignant gastrointestinal diseases (44 patients with colon adenocarcinoma, 21 with gastric cancer, 15 with hepatocellular adenocarcinoma, 10 patients with pancreas adenocarcinoma and eight patients with esophagus cancer). The serum levels of carotenoids were measured with high-pressure liquid chromatography. The sera of the patients were taken at the time of the diagnosis. RESULTS: the measurements indicated that (1) the serum level of vitamin A and zeaxanthin were significantly lower in all of these groups (except of pancreas adenocarcinoma), but the extent of the A decrease was different in the patients with different types of gastrointestinal malignancy. The serum level of vitamin A was in the healthy subjects 2.072+/-0.332 mmol/l and in the case of gastrointestinal malignancies was 0.77+/-0.14 mmol/l (P<0.001) The serum level of zeaxanthin was in the healthy subjects 0.143+/-0.057 mmol/l and at the malignancies was 0.042+/-0.014 mmol/l (P<0.01). (2) There were no significant differences in the serum levels of other carotenoids in the checked groups. (3) The serum level of cholesterol, total protein, albumin and haemoglobin were in the normal range in these patients. These results indicate that the carotenoids may be responsible nutritional factors (as nutritional scavengers) in the development of different malignant diseases. This supposed role in the carcinogenesis does not depend fully on the vitamin A activity.