Identification of significant regions of transcription factor DP-1 (TFDP-1) involved in stability/instability of the protein

We previously reported the identification of DP-1 isoforms (α and β), which are structurally C-terminus-deleted ones, and revealed the low-level expression of these isoforms. It is known that wild-type DP-1 is degraded by the ubiquitin-proteasome system, but few details are known about the domains concerned with the protein stability/instability for the proteolysis of these DP-1 isoforms. Here we identified the domains responsible for the stability/instability of DP-1. Especially, the DP-1 “Stabilon” domain was a C-terminal acidic motif and was quite important for DP-1 stability. Moreover, we propose that this DP-1 Stabilon may be useful for the stability of other nuclear proteins when fused to them.     

DNA Sequences from Formalin-Fixed Nematodes: Integrating Molecular and Morphological Approaches to Taxonomy

To effectively integrate DNA sequence analysis and classical nematode taxonomy, we must be able to obtain DNA sequences from formalin-fixed specimens. Microdissected sections of nematodes were removed from specimens fixed in formalin, using standard protocols and without destroying morphological features. The fixed sections provided sufficient template for multiple polymerase chain reaction-based DNA sequence analyses.     

酸中毒条件下脓毒性休克大鼠胸主动脉对多巴胺反应性的变化

目的:观察不同酸中毒条件下正常大鼠和脓毒性休克大鼠胸主动脉对多巴胺反应性的变化。方法:采用离体血管灌流方法,观察对照组和脓毒性休克组大鼠胸主动脉在不同pH条件下的反应性变化。结果:pH值依次降低,对照组及脓毒性休克组离体胸主动脉对多巴胺反应性均下降,在相同pH值条件下脓毒性休克组比对照组离体血管对多巴胺反应性下降更为明显。结论:①环境pH值的下降会导致正常大鼠和脓毒性休克大鼠离体动脉对多巴胺反应性的下降。②在相同的酸性环境中脓毒性休克大鼠的血管对多巴胺刺激的反应性更差,更易失去活性。     

Midsummer heat wave effects on lacustrine plankton: Variation of assemblage structure and fatty acid composition

The effects of the 2003 European heat wave on a freshwater plankton assemblage and its fatty acid (FA) composition were investigated. Composition and FA profiles of four size categories of planktonic organisms collected in 2003 were compared to those of the colder year 2002.     

Photosynthetic pigment concentration, organization and interconversions in a pale green Syrian landrace of barley (Hordeum vulgare L., Tadmor) adapted to harsh climatic conditions

Tadmor is a Syrian barley landrace that has adapted to semi-arid environments. Its leaves are pale green because of a 30% decrease in the chlorophyll and the carotenoid content of the chloroplasts (leading to a 7·5% decrease in light absorption) compared with barley genotypes that are not adapted to harsh Mediterranean climatic conditions (e.g. Plaisant). This difference in pigment content was attenuated during growth of the plants in strong light, but was strongly amplified when strong light was combined with a high growth temperature. The low pigment content of Tadmor leaves was not associated with significant changes in the pigment distribution between the photosystems or between the reaction centres of the photosystems and their associated chlorophyll antennae. No significant difference in the photosynthetic activity (O₂ production per unit absorbed light) was observed between Tadmor and Plaisant. The conversion of violaxanthin to zeaxanthin in strong light and its reversal in darkness were much faster and operated at a higher capacity in Tadmor leaves compared with Plaisant leaves, resulting in an increased photostability of photosystem II in the former leaves. The accelerated xanthophylls interconversion in the Syrian landrace was associated with, and possibly related to, an increased fluidity of the thylakoid membranes. The lipid peroxide level was lower in Tadmor compared with Plaisant. In contrast, no difference was found in the non-photochemical quenching of chlorophyll fluorescence between the two barley genotypes. The data indicate that the pale green Syrian landrace is equipped to survive excessive irradiance through a passive reduction of the light absorptance of its leaves, which mitigates the heating effects of strong light, and through the active protection of its photochemical apparatus by a rapid xanthophyll cycling.     

The Detection of Hamster Connexins: A Comparison of Expression Profiles with Wild-Type Mouse and the Cancer-Prone Min Mouse

The open reading frames of 17 connexins from Syrian hamster (using tissues) and 16 connexins from the Chinese hamster cell line V79, were fully (Cx30, Cx31, Cx37, Cx43 and Cx45) or partially sequenced. We have also detected, and partially sequenced, seven rat connexins that previously were unavailable. The expression of connexin genes was examined in some hamster organs and cultured hamster cells, and compared with wild-type mouse and the cancer-prone Min mouse. Although the expression patterns were similar for most organs and connexins in hamster and mouse, there were also some prominent differences (Cx29 and 30.3 in testis; Cx31.1 and 32 in eye; Cx46 in brain, kidney and testis; Cx47 in kidney). This suggests that some connexins have species-specific expression profiles. In contrast, there were minimal differences in expression profiles between wild type and Min mice. Species-specific expression profiles should be considered in attempts to make animal models of human connexin-associated diseases.     

Regeneration of Triticum aestivum apical explants after microinjection of germ line progenitor cells with DNA

A highly efficient method of regenerating fertile, phenotypically normal plants from shoot apex cultures of T. aestivum was developed. The hypodermal layer (L2) of the vegetative apex containing germ line precursor cells could be located with bright field microscopy and targeted for microinjection. Fluorescently labelled dextrans were used as markers to develop a microinjection procedure which did not disrupt nuclear or cytoplasmic structure. This procedure was used to inject plasmid DNA into L2 cells. Capillary microinjection did not shear the plasmid DNA and delivery of DNA was confirmed by polymerase chain reaction analysis of DNA isolated from injected apices. The significance of these findings in relation to the development of cereal transformation systems will be discussed.     

DPHL:A DIA Pan-human Protein Mass Spectrometry Library for Robust Biomarker Discovery

To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipeline and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to generate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.     

Hyperthermia Induces the Synthesis of a Heat Shock Protein by Polysomes Isolated from the Fetal and Neonatal Mammalian Brain

Abstract: Analysis of the cell-free translation products of polysomes isolated from fetal brain and other organs indicates that elevation of maternal body temperature induces the synthesis of a heat shock protein of molecular weight 74,000 (74K). The newborn mammal is particularly sensitive to induction of the 74K protein. As early postnatal development proceeds, higher body temperatures are required to induce synthesis of the 74K heat shock protein.