Effect of dexamethasone on kininogen production by a rat hepatoma cell line

T Kininogen and High Molecular Weight Kininogen were characterized in the cell culture medium of Fao cells, a highly differentiated cell line derived from the Reuber H35 rat hepatoma. Immunoreactive T Kininogen and High Molecular Weight Kininogen identified by direct and specific RIAs were indistinguishable from standard kininogens. Immunoreactive T Kininogen was further identified by HPLC analysis of T kinin released after trypsin hydrolysis of the cell culture medium. The basal release rate of T kininogen was ten-fold higher than that of High Molecular Weight Kininogen. T Kininogen was not stored within the cells contrary to High Molecular Weight Kininogen. The production of the two kininogens in the cell medium was stimulated by dexamethasone up to five times in a dose-dependent manner. The specific antiglucocorticoid compound RU 38486 did not alter the basal rate of kininogen release by Fao cells, but abolished the stimulation by dexamethasone, indicating that dexamethasone exerts a true glucocorticoid type effect.     

DARPin Ec1-LMWP protein scaffold in targeted delivery of siRNA molecules through EpCAM cancer stem cell marker

Molecular Biology Reports - This study is to investigate the binding ability of Designed Ankyrin Repeat Proteins type Ec1that was fused to Low Molecular Weight Protamine (DARPin Ec1-LMWP) protein...     

Purification to homogeneity and biochemical characterization of two suppressor factors from human malignant T-cells

A cell clone (GI-CO-T-9) derived from a long term T-cell culture (PF-382), established from a patient affected by acute T-lymphoblastic leukemia (T-ALL), was selected for the presence in the culture medium of factors suppressing T-cell proliferation. The crude supernatant has been subjected to a multi-step chromatographic fractioning, including: preparative gel permeation, anion exchange, and hydrophobic interaction High Performance Liquid Chromatography (HPLC). The highly purified material was characterized by polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), revealing single bands of 115 Kd and 80 Kd. The isoelectric points (pI), determined by flat-bed isoelectric-focusing, were 7.4 for High Molecular Weight Suppressor Factor (HMWSF) and 3.5-3.6 for Low Molecular Weight Suppressor Factor (LMWSF).     

Radioimmunoassay of low molecular weight human kininogen

A radioimmunoassay for low molecular weight (LMW) human Kininogen has been carried out. The first step was to prepare LMW Kininogen from human plasma. The proposed method allowed to get chemically pure and biologically active LMW Kininogen. This preparation was used to induce antibody. Optimal conditions for labelling and incubation were determined. This method may be applied to the assay of Kininogen in human plasma.     

Soils of the Chinese Hubei province show a very high diversity of Sinorhizobium fredii strains

Biodiversity studies of native soybean-nodulating rhizobia in soils from the Chinese Hubei province (Honghu county; pH 8, alluvial soil) have been carried out. Inoculation of an American (Williams) and an Asiatic (Peking) soybean cultivar with eleven soil samples led to the isolation of 167 rhizobia strains. The ratio (%) of slow-/fast-growing isolates was different depending on the trap plant used. All isolates were able to nodulate both cultivars, although the N2-fixation efficiency (measured as plant-top dry weight) was different among them. A total of thirty-three isolates were selected for further characterisation on the basis of physiological parameters, PCR-RFLP of symbiotic genes and Low Molecular Weight RNA, lipopolysaccharide, protein and plasmid profiles. Low Molecular Weight RNA profiling indicates that all the isolates belong to species Sinorhizobium fredii. The dendrogram obtained with the physiological parameters has been useful to classify the isolates at strain level, although plasmid profiling was the most discriminating technique to detect differences among the analysed soybean-rhizobia isolates, showing there is not two isolates identical each other. Plasmid profile analyses also revealed that some of the investigated strains contain low molecular weight plasmids (7-8-kb). They are, to our knowledge, the smallest ever found in rhizobia and they could be the starting point for the construction of the first group of vectors based on a native rhizobia replicon.     

SAR of non-hydrolysable analogs of pyridoxal 5′-phosphate against low molecular weight protein tyrosine phosphatase isoforms

Kinases and phosphatases are key enzymes in cell signal transduction pathways. Imbalances in these enzymes have been linked to numerous disease states ranging from cancer to diabetes to autoimmune disorders. The two isoforms (IFA and IFB) of Low Molecular Weight Protein Tyrosine Phosphatase (LMW-PTP) appear to play a role in these diseases. Pyridoxal 5′-phosphate (PLP) has been shown to act as a potent but, impractical micromolar inhibitor for both isoforms. In this study, a series of non-hydrolysable phosphonate analogs of PLP were designed, synthesized and tested against the two isoforms of LMW-PTP. Assay results demonstrated that the best inhibitor for both isoforms was compound 5 with a K_is of 1.84 μM (IFA) and 15.6 μM (IFB). The most selective inhibitor was compound 16, with a selectivity of roughly 370-fold for IFA over IFB.     

Organic and inorganic human-induced contamination of Posidonia oceanica meadows

In coastal environments, plants are used for phytoremediation of contamination. Organic and inorganic contaminants may be due to natural and/or anthropogenic sources. The aim of this study is to compare inorganic (trace metal) and organic (PAH) contamination in Posidonia oceanica and to analyse the relationship between these types of pollutants indeed very few studies have been interested in their correlations and common sources. P. oceanica leaves were collected in two sites exhibiting different levels of human-induced pressure. Higher values were recorded in the more polluted site (Toulon) for trace metals (Ag, Hg, Pb) as well as for PAHs (Medium Molecular Weight and High Molecular Weight) due to the presence of the city and/or harbour in proximity. For the first time in a coastal environment, correlations were observed between metals and PAHs.     

Platelet-dependent signaling and Low Molecular Weight Protein Tyrosine Phosphatase expression promote aggressive phenotypic changes in gastrointestinal cancer cells

Over the last decades, some members of the protein tyrosine phosphatase family have emerged as cancer promoters. Among them, the Low Molecular Weight Protein Tyrosine Phosphatase (LMWPTP) has been described to be associated with colorectal cancer liver metastasis and poor prostate cancer prognosis. Of importance in the process of cancer progression and metastasis is the interaction between tumor cells and platelets, as the latter are thought to promote several tumor hallmarks. Here, we examine to what extent LMWPTP expression in tumor cells affects their interaction with platelets. We demonstrate that the gene encoding LMWPTP is overexpressed in upper gastrointestinal (GI) cancer cell as well as colorectal cancer, and subsequently employ cell line models to show that the level of this phosphatase may be further augmented in the presence of platelets. We demonstrate that tumor-platelet interaction promotes GI tumor cell proliferation. Additionally, using know-down/-out models we show that LMWPTP expression in cancer cells contributes to a more efficient interaction with platelets and drives platelet-induced proliferation. These data are the first to demonstrate that phosphatases play a positive role in the tumor-promoting activities of platelets, with LMWPTP emerging as a key player promoting oncogenic phenotypic changes in tumor cells.     

Screening the low molecular weight fraction of human serum using ATR‐IR spectroscopy

Vibrational spectroscopic techniques can detect small variations in molecular content, linked with disease, showing promise for screening and early diagnosis. Biological fluids, particularly blood serum, are potentially valuable for diagnosis purposes. The so‐called Low Molecular Weight Fraction (LMWF) contains the associated peptidome and metabolome and has been identified as potentially the most relevant molecular population for disease‐associated biomarker research. Although vibrational spectroscopy can deliver a specific chemical fingerprint of the samples, the High Molecular Weight Fraction (HMWF), composed of the most abundant serum proteins, strongly dominates the response and ultimately makes the detection of minor spectral variations a challenging task. Spectroscopic detection of potential serum biomarkers present at relatively low concentrations can be improved using pre‐analytical depletion of the HMWF. In the present study, human serum fractionation by centrifugal filtration was used prior to analysis by Attenuated Total Reflection infrared spectroscopy. Using a model sample based on glycine spiked serum, it is demonstrated that the screening of the LMWF can be applied to quantify blinded concentrations up to 50 times lower. Moreover, the approach is easily transferable to different bodily fluids which would support the development of more efficient and suitable clinical protocols exploring vibrational spectroscopy based ex‐vivo diagnostic tools.

Revealing serum LMWF for spectral serological diagnostic applications.     

Transcriptomic Analyses of Chilling Stress Responsiveness in Leaves of Tobacco (Nicotiana tabacum) Seedlings

Plant Molecular Biology Reporter - Low temperature is among the most significant abiotic stresses restricting geographical distribution of plants and reducing crop productivity. However, the...     

5-Aza-2’-deoxycytidine induces a greater inflammatory change,at the molecular levels,in normoxic than hypoxic tumor microenvironment

Molecular Biology Reports - Hypoxia is associated with tumor aggressiveness and poor prognosis, including breast cancer. Low oxygen levels induces global genomic hypomethylation and...     

Increasing the colon cancer cells sensitivity toward radiation therapy via application of Oct4–Sox2 complex decoy oligodeoxynucleotides

Molecular Biology Reports - Low sensitivity of cancer stem cells toward regular cancer therapy strategies is an important issue in the field of cancer remedy. The concept of cancer stem cell...     

Molecular mechanisms and future improvement of submergence tolerance in rice

Molecular Breeding - Low temperature (LT) is a key environmental stress affecting the growth, development, yield, and quality of wheat (Triticum aestivum L.). To better understand the genetic basis...     

Coenzyme Q10 in association with metabolism-related AMPK/PFKFB3 and angiogenic VEGF/VEGFR2 genes in breast cancer patients

Molecular Biology Reports - Low levels of coenzyme Q10 (CoQ10) have been reported in the circulation of patients with breast cancer, particularly in metastatic features. Our objective was to study...     

Activation of the Low Molecular Weight Protein Tyrosine Phosphatase in Keratinocytes Exposed to Hyperosmotic Stress

Herein, we provide new contribution to the mechanisms involved in keratinocytes response to hyperosmotic shock showing, for the first time, the participation of Low Molecular Weight Protein Tyrosine Phosphatase (LMWPTP) activity in this event. We reported that sorbitol-induced osmotic stress mediates alterations in the phosphorylation of pivotal cytoskeletal proteins, particularly Src and cofilin. Furthermore, an increase in the expression of the phosphorylated form of LMWPTP, which was followed by an augment in its catalytic activity, was observed. Of particular importance, these responses occurred in an intracellular milieu characterized by elevated levels of reduced glutathione (GSH) and increased expression of the antioxidant enzymes glutathione peroxidase and glutathione reductase. Altogether, our results suggest that hyperosmostic stress provides a favorable cellular environment to the activation of LMWPTP, which is associated with increased expression of antioxidant enzymes, high levels of GSH and inhibition of Src kinase. Finally, the real contribution of LMWPTP in the hyperosmotic stress response of keratinocytes was demonstrated through analysis of the effects of ACP1 gene knockdown in stressed and non-stressed cells. LMWPTP knockdown attenuates the effects of sorbitol induced-stress in HaCaT cells, mainly in the status of Src kinase, Rac and STAT5 phosphorylation and activity. These results describe for the first time the participation of LMWPTP in the dynamics of cytoskeleton rearrangement during exposure of human keratinocytes to hyperosmotic shock, which may contribute to cell death.     

Speciation and Environmental Fate of Chromium in Rivers Contaminated with Tannery Effluents

Redox and size speciation of chromium in rivers contaminated with tannery wastewater was carried out to provide insight into its transport and removal mechanisms. Total chromium was determined with Inductively Coupled Plasma‐Mass Spectrometry and Cr (VI) with Catalytic Adsorption Stripping Voltammetry. For the size speciation, particles were retained with a cartridge filter (cut‐off 1.2 μm) and the total filterable fraction was further fractionated with Tangential Flow Filtration to determine the concentrations of chromium associated with the High Molecular Weight Colloidal (HMWC), Low Molecular Weight Colloidal (LMWC) and Truly Dissolved (TD) fractions. Two fluvial systems of similar sizes, but located in contrasting climatic zones, were selected for comparison: the Sebou‐Fez system in Morocco and Dunajec River‐Czorsztyn Reservoir system in Poland. Particulate Cr dominated in the Sebou‐Fez system (about 90 %); while in the Dunajec‐Czorsztyn system, it represented only 17–53 % of the total chromium in raw water. Still, the partition coefficients [K_d] were of the same magnitude. Chromium (III) was the only form detected in Sebou‐Fez, whereas in Dunajec‐Czorsztyn Cr (VI) was also present with its proportion increasing downstream from the input of tannery wastewater due to the preferential removal of Cr (III). In the filtered water in Morocco a large fraction of Cr occurred in the HMWC fraction (50–70 %) at the two most contaminated sites, while the LMWC and TD forms prevailed at the non‐contaminated sites in the Sebou River. At a very high concentration, in the water in the proximity of tanneries (well above the theoretical saturation level) Cr precipitated as polynuclear Cr‐hydroxide. In Dunajec‐Czorsztyn, the partition of Cr (III) was approximately equal between the HMWC, LMWC and TD fractions, in contrast to Cr (VI) which occurred almost exclusively in the TD fraction. In both systems, Cr (III) was rapidly removed from the water to the sediments. The confluence of the Sebou with the Fez and the Czorsztyn reservoir trapped efficiently Cr (III) preventing its spreading over long distances. Cr (VI) showed conservative behavior and bypassed the Czorsztyn Reservoir. This study provides a first set of data on the partitioning of Cr (III) and Cr (VI) between the particulate, the colloidal and truly dissolved fractions in fluvial systems contaminated with tannery effluents. It also suggests that, in these systems, truly dissolved Cr (III) can be adequately modeled from the total filterable concentrations.     

小麦HMW谷蛋白亚基基因克隆研究进展

高分子量麦谷蛋白亚基 (HMW GS)作为小麦胚乳中的重要贮藏蛋白 ,其组成及含量对小麦面粉的烘烤品质具有重要的决定作用。因此 ,改变小麦中HMW 谷蛋白的组成及含量是小麦品质改良的主要内容。而定向克隆小麦HMW GS基因则为利用基因工程方法改良小麦品质提供新的基因资源 ,从而为优质小麦的发展起到积极的推动作用。综述了近 2 0年来国内外小麦HMW GS基因克隆的研究进展 ,并讨论了近年来发展起来的一些新的基因克隆方法及其在小麦HMW GS基因克隆上的应用前景。     

The double-edged sword of Nrf2: subversion of redox homeostasis during the evolution of cancer

Low levels of Nrf2 activity predispose cells to chemical carcinogenesis. Surprisingly, Padmanabhan et al. (2006) provide evidence in a recent issue of Molecular Cell to support the notion that elevated Nrf2 activity may also play a role in the evolution of cancer.