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PubMed2004

Time-resolved chemiluminescence technique for the microdetermination of proteins based on their complexation with copper(II).

Based on the complexation between proteins and Cu(II) coupled with the time-resolved chemiluminescence (CL) technique, a highly sensitive and quantitative assay for measuring proteins in solution is described. The complexes of proteins with Cu(II) have a strongly catalytic effect on the luminol-H2O2 CL reaction. Because the CL emission produced by the complexes is much more long-lived than that by Cu(II), the CL signals originating from proteins can be easily identified and measured with a time-resolved technique. On this basis, bovine albumin fraction V (BAF V) can be quantitatively determined in the range of 0.01 - 5.0 microg/ml with a detection limit of 5.8 ng/ml. The results show that the proposed assay exhibits a small variation in the response values for the same amount of different proteins, as compared to the Lowry as well as Bradford assays. The CL assay has also been studied for the detection of immobilized proteins.

Calibration↗

PubMed2006

Aeration control for simultaneous nitrification-denitrification in a biological aerated filter using internal model approach.

Experience has shown that simultaneous nitrification/denitrification in a biological aerated filter is possible. However these systems react very sensitively to the aeration control strategies applied. Poorly adapted control strategies induce a strong decrease in treatment efficiency. A new control strategy for simultaneous N/DN is developed. The strategy proposed makes use of ammonia measurements and the inlet and outlet: a Feedback/Feedforward block. Passing by a calculation of the load to be eliminated, an estimation of the air flow velocity to be applied is carried out dynamically. A retroactive loop corrects this prediction in order to reach exactly the desired set point. This control approach has been implemented and tested at pilot plant scale for a period of 18 months. The pilot plant consists of two coupled BAF cells, reflecting closely industrial scale situations. Comparative studies reveal clearly the improved performance of the developed FF/FB control strategy compared to classical controllers. The benefits include 5% increase in nitrogen removal performance and a reduction of 15 to 20% in air requirement, offering a rapid return of investment costs.

Algorithms↗

PubMed2006

Utilization of dried sludge for making ceramsite.

Dried sludge as additive for making ceramsite is a new effective approach for disposal of sludge. In this study sewage sludge, water glass and clay were chosen as the components, the optimal ratio of the components and the most appropriate conditions were obtained. The functions of primary components in the sintering process, porosity formation mechanism and solid phase reaction also have been discussed. The optimized process parameters were shown as follows: the ratio of dried sludge/clay (wt%) was 33%, ratio of adherent /clay (wt%) was 15%, sintering temperature was 1000 degrees C, sintering time was 10 min. Bulk density was 582 kg m(-3), particle density was 1,033 kg m(-3), water absorption was 9.5%, porosity was 43.7%. SEM, EDS, XRD and XRF analyses were also carried out. The results indicate that dried sludge as raw material is a good way for making ceramsite. Biological Aerated Filters (BAFs) with filter media of Guangzhou ceramsite, Jiangxi ceramsite, activated carbon and ceramsite (obtained in test) were selected to treat municipal wastewater. The average removal efficiencies of ceramsite (obtained in test) for turbidity, COD, SCOD and NH3-N were about 96.4%, 76.2%, 59.6% and 82.3% respectively and were higher than those of other ceramsites.

Absorption↗

PubMed1999

Influence of prolactin on the differentiation of mouse B-lymphoid precursors.

Development and activation of immune cells are submitted to hormonal influences, as illustrated by the roles of corticosteroids in thymus, pregnancy-related estrogens in B-cell development, or prolactin (PRL) on T-cell generation and function. We have analyzed the putative role of PRL in B lymphopoiesis and differentiation. We chose as an experimental model the interleukin (IL)-3 dependent BaF-3 pro-B cell line, which was transfected with the rat long form of the PRL receptor (PRL-R) and transferred from IL-3- to PRL-enriched media. When stimulated with PRL, the PRL-R transfectants underwent some changes characteristic of B-cell differentiation: (a) IL-2R alpha chain became positively controlled by PRL; (b) antiapoptotic Bcl-2 protein was induced by PRL in a dose-dependent manner; and (c) transcription of the pre-B cell receptor encoding the lambda5 gene was strongly up-regulated. We attempted to evaluate the differentiation-promoting activity of PRL in more physiological conditions, and the presence of PRL-R in bone marrow B-cell precursors was revealed. Furthermore, PRL promoted significant expansions of defined B-lineage cell populations in short-term bone marrow cell cultures. These findings suggest that PRL, in collaboration with other cytokines and hormonal influences, modulates B-cell development.

Animals↗

PubMed1999

[The detection of apoptosis cells by TUNEL stain in peripheral blood smears].

We have determined the fixative conditions for TUNEL stain in peripheral blood smears. Murine IL-3 dependent B cell line (BAF-3) was used as positive control. The intensity of fixing solutions effected on the positivity/negativity for TUNEL stain. Alive cells showed positivity for TUNEL stain (false positive), when 70% methyl alcohol, acetone or carnoy fluid was used. In contrast to the strong fixation by high concentrated glutaraldehyde and the phosphate buffer, denatured cells showed to be negative (false negative). The fixation by 0.4% glutaraldehyde and 70% methyl alcohol for 15 min at room temperature, followed by the treatment by 20 micrograms/ml proteinase K for 15 min at 37 degrees C was most suitable for TUNEL stain to detect the apoptotic cells in peripheral blood smears. The frequency of TUNEL positive cells increased significantly in peripheral blood cells treated by apoptosis induced reagents, or infected by CMV. This is a new approach for the techniques to detect the apoptotic cells in peripheral blood smears.

Apoptosis↗

PubMed2000

Different mechanisms define the antiadhesive function of high molecular weight kininogen in integrin- and urokinase receptor-dependent interactions.

Proteolytic cleavage of single-chain high molecular weight kininogen (HK) by kallikrein releases the short-lived vasodilator bradykinin and leaves behind 2-chain high molecular weight kininogen (HKa) that has been previously reported to exert antiadhesive properties as well as to bind to the urokinase receptor (uPAR) on endothelial cells. In this study we defined the molecular mechanisms for the antiadhesive effects of HKa related to disruption of integrin- and uPAR-mediated cellular interactions. Vitronectin (VN) but not fibrinogen or fibronectin-dependent alphavbeta(3) integrin-mediated adhesion of endothelial cells was blocked by HKa or its isolated domain 5. In a purified system, HKa but not HK competed for the interaction of VN with alphavbeta(3) integrin, because HKa and the isolated domain 5 but not HK bound to both multimeric and native VN in a Zn(2+)-dependent manner. The interaction between HKa or domain 5 with VN was prevented by heparin, plasminogen activator inhibitor-1, and a recombinant glutathione-S-transferase (GST)-fusion peptide GST-VN (1-77) consisting of the amino terminal portion of VN (amino acids 1-77), but not by a cyclic arginyl-glycyl-aspartyl peptide, indicating that HKa interacts with the amino terminal portion of VN ("somatomedin B region"). Furthermore, we have confirmed that HKa but not HK bound to uPAR and to the truncated 2-domain form of uPAR lacking domain 1 in a Zn(2+)-dependent manner. Through these interactions, HKa or its recombinant His-Gly-Lys-rich domain 5 completely inhibited the uPAR-dependent adhesion of myelomonocytic U937 cells and uPAR-transfected BAF-3 cells to VN and thereby promoted cell detachment. By immunogold electron microscopy, both VN and HK/HKa were found to be colocalized in sections from human atherosclerotic coronary artery, indicating that the described interactions are likely to take place in vivo. Taken together, HK and HKa inhibit different VN-responsive adhesion receptor systems and may thereby influence endothelial cell- or leukocyte-related interactions in the vasculature, particularly under inflammatory conditions. (Blood. 2000;96:514-522)

Arteriosclerosis↗

PubMed2000

Histone deacetylase inhibitors suppress IL-2-mediated gene expression prior to induction of apoptosis.

Histone deacetylase (HDAC) inhibitors can induce transcriptional activation of a number of genes and induce cellular differentiation as histone acetylation levels increase. Although these inhibitors induce apoptosis in several cell lines, the precise mechanism by which they do so remains obscure. This study shows that HDAC inhibitors, sodium butyrate and trichostatin A (TSA), abrogate interleukin (IL)-2-mediated gene expression in IL-2-dependent cells. The HDAC inhibitors readily induced apoptosis in IL-2-dependent ILT-Mat cells and BAF-B03 transfectants expressing the IL-2 receptor betac chain, whereas they induced far less apoptosis in cytokine-independent K562 cells. However, these inhibitors similarly increased acetylation levels of histones in both cells. Although histone hyperacetylation is believed to lead to transcriptional activation, the results showed an abrogation of IL-2-mediated induction of c-myc, bag-1, and LC-PTP gene expression. This observed abrogation of gene expression occurred prior to phosphatidylserine externalization, a process that occurs in early apoptotic cells. Considering the biologic role played by IL-2-mediated gene expression in cell survival, these data suggest that its abrogation may contribute to the apoptotic process induced by HDAC inhibitors. (Blood. 2000;96:1490-1495)

Animals↗

PubMed2001

Reed bed dewatering of agricultural sludges and slurries.

In trials at Rugeley, UK, reed beds were used for dewatering agricultural sludges and slurries. Three beds, each of 3.5 m2, were employed, two planted with Phragmites australis, the third left unplanted as control. The sludge contained partly oxidised solids from a Biological Aerated Filter (BAF) treating weak pig slurry. It was supplemented with untreated settled pig slurry. Following reed establishment planted Bed A was fed at a constant similar rate to the unplanted Control Bed C. The second planted Bed B was fed at higher rates alternating with rest periods. On this bed the aeration pipes were blocked off. The trials were run for 16 months, which included two summer periods. The results showed that the planted Bed A had definitely better dewatering ability than the unplanted one fed at a similar rate. During the summer months Bed B could be fed at over twice the rate used for the constant input beds. The percolate from the control bed was more highly oxidised than from the planted beds, probably due to a longer holdup time in the absence of reeds. On Bed B the reed quality deteriorated during the second year, after use of untreated slurry as feed.

Agriculture↗

PubMedDate not supplied

[Imidazoline receptors-normal and pathological factors].

Imidazolinic receptors I1 and I2A and I2B different from alpha 2 receptors and by any other type of receptors were identified in the last years in many tissues from human body and in cells of a great number of animals. There are agonists and antagonists on I1 receptors (e.g., agonists like moxonidine, rilmenidine and antagonists like efaroxan, BAF 6143 etc.) and also on I2 receptors (e.g., agonist-idazoxan, etc.). There are many actions performed by stimulation of imidazolinic receptors like: decreasing of blood pressure (I1 stimulation from midbrain), decreasing of noradrenaline plasmatic level, antiarrhythmic actions in experimental arrhythmias etc. Imidazolinic receptors are involved in natriuresis, pancreatic insulin production, some superior nervous activities, regulation of food intake etc. We consider that imidazolinic receptors and their endogenous agonists (e.g. agmatine) represent an important system involved in modulation of cell function.

Adrenergic Agonists↗

PubMed2002

Polycyclic aromatic hydrocarbon bioaccumulation by meiobenthic copepods inhabiting a superfund site: techniques for micromass body burden and total lipid analysis.

Microtechniques for polycyclic aromatic hydrocarbon (PAH) body burden and total lipid analysis were developed and applied to determine the first lipid-normalized bioaccumulation factors for a hydrophobic organic toxicant in a meiobenthic organism (0.063-0.500 mm) living in field-contaminated sediments. The total lipid microtechnique combines the standard Bligh-Dyer extraction method with a colorimetric quantification method for analysis of samples containing I to 50 microg lipid. The microtechnique for body burden analysis quantifies PAHs from tissue samples containing as little as 10 pg PAH. Fluoranthene, benz[a]anthracene, and benzo[a]pyrene biota-sediment accumulation factors (BSAFs) were determined for the meiobenthic copepod Microarthridion littorale living in an estuarine U.S. Environmental Protection Agency Superfund site. Gravid female, nongravid female, and male BSAFs were 0.82, 0.54, and 0.36, respectively, for fluoranthene; 0.50, 0.44, and 0.40, respectively, for benz[a]anthracene; and 0.09, 0.12, and 0.15, respectively, for benzo[a]pyrene. Comparison of nonlipid-normalized bioaccumulation factors (BAFs) to BSAFs indicates that M. littorale bioaccumulated PAHs on a gram lipid basis. The BSAFs declined consistently with increasing PAH log K(ow) for all copepod sex and reproductive stages. Sex- and stage-specific comparisons of BSAFs suggest that differences in lipid content and quality may lead to differences in BSAF values depending on PAH molecular weight and/or hydrophobicity.

Animals↗

PubMed2002

Biological availability of traffic-related platinum-group elements (palladium, platinum, and rhodium) and other metals to the zebra mussel (Dreissena polymorpha) in water containing road dust.

The uptake and bioaccumulation of 15 road dust metals by the zebra mussel (Dreissena polymorpha) were investigated in laboratory exposure studies with emphasis on the traffic-related platinum-group elements (PGEs) palladium (Pd), platinum (Pt), and rhodium (Rh). The biological availability of the metals may depend on water characteristics, so the mussels were maintained in two types of water: nonchlorinated tap water and humic water of a bog lake, both of which contained dust of a moderately frequented road. After an exposure period of 26 weeks, soft tissues of the mussels were freeze-dried and analyzed for the metals. The metal concentrations in the mussel soft tissue ranged from several hundred micrograms per gram (e.g., for iron [Fe]) to less than 10 ng/g (for PGEs). Metal uptake from the road dust by the mussels was found for the PGEs and silver (Ag), bismuth (Bi), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), Fe, lead (Pb), and antimony (Sb). After maintenance of mussels in road dust-contaminated tap water, bioaccumulation factors (BAF = (C(exposed mussels) - C(control mussels))/C(total metal, water), where c is concentration) decreased in the following order: Cu > Cd > Ag > Pd > Sb > Pb > Fe > Pt > Rh. The biological availability of most metals was enhanced by humic water as compared to tap water. Our results show a hitherto unrecognized high availability of Pd for the mussels. Thus, this metal should be monitored more intensively in the environment to assess its distribution in the biosphere.

Animals↗

PubMed2002

Functional cross-talk between cytokine receptors revealed by activating mutations in the extracellular domain of the beta-subunit of the GM-CSF receptor.

Several reports have suggested an interaction between the erythropoietin receptor (EpoR) and the shared signaling subunit (hbeta(c)) of the human granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-3, and IL-5 receptors, although the functional consequences of this interaction are unclear. We previously showed that in vivo expression of constitutively active extracellular (EC) mutants of hbeta(c) induces erythrocytosis and Epo independence of erythroid colony-forming units (CFU-E). This occurs despite an apparent requirement of these mutants for the GM-CSF receptor alpha-subunit (GMRalpha), which is not expressed in CFU-E. Here, we show that coexpression of hbeta(c) EC mutants and EpoR in BaF-B03 cells, which lack GMRalpha, results in factor-independent proliferation and JAK2 activation. Mutant receptors that cannot activate JAK2 fail to produce a functional interaction. As there is no detectable phosphorylation of hbeta(c) on intracellular tyrosine residues, EpoR displays constitutive tyrosine phosphorylation. These observations suggest that JAK2 activation mediates cross-talk between EC mutants of hbeta(c) and EpoR. The implications of these data are discussed as are our findings that activated hbeta(c) mutants can functionally interact with certain other cytokine receptors.

Animals↗

PubMed2002

Evaluation of estrogen-like activity on sewage treatment processes using recombinant yeast.

Four sewage treatment plants based on an activated sludge process and a pilot scale plant for advanced sewage treatment located in Japan were evaluated for removal of estrogenic substances using in vitro recombinant yeast assay and chemical analysis. The results indicated that 17beta-estradiol (E2) significantly contributed to estrogen-like activity analyzed by yeast assay especially in secondary treated effluents. On the other hand, batch study showed that estrogen-like activity of spiked E2 was easily decreased by an activated sludge treatment. This result suggested that E2 concentrations measured by enzyme immunoassay (EIA) were interpreted as false positives in effluents, and that unknown estrogenic substances other than E2 might have contributed to estrogen-like activity in the secondary treated effluents. Further, in the pilot scale study, advanced sewage treatment processes such as a biological aerated filtration (BAF) process, an advanced oxidation process (AOP), were effective for the removal of those estrogenic activities contributed by unknown estrogenic substances in sewage secondary treated effluent.

Endocrine System↗

PubMed2003

Inactivation of Salmonella spp. from secondary and tertiary effluents by UV irradiation.

The aim of this study was to verify the efficiency of UV irradiation in the inactivation of Salmonella spp. in treated wastewater with different levels of turbidity and exposed to increasing doses of UV irradiation. Experiments were carried out in a batch reactor and in a real scale reactor. Salmonellae obtained from clinical samples were seeded into autoclaved wastewater collected from a wastewater treatment plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The results showed that salmonellae were not inactivated in effluents from the UASB reactor indicating that the presence of suspended solids was an important obstacle to UV penetration in bacteria. However, UV irradiation was efficient in inactivating Salmonella of effluents from aerated secondary and tertiary biofilm reactors.

Biofilms↗

PubMed2003

Temperature effect on aerobic denitrification and nitrification.

Nitrogen loss without organic removal in biofilter was observed and its possible reason was explained. A lower hydraulic loading could improve aerobic denitrification rate. Aerobic denitrification was seriously affected by low temperature(below 10 degrees C). However, nitrification rate remained high when the temperature dropped from 15 degrees C to 5 degrees C . It seemed the autotrophic biofilm in BAF could alleviate the adverse effect of low temperature.

Biodegradation, Environmental↗

PubMed2004

Experimental study on nitrification in a submerged aerated biofilter.

The aim of the present work was to evaluate the performance of a semi-pilot scale BAF in order to obtain a highly polished effluent in terms of removal of organic matter, suspended solids and ammonia and to observe the influence of temperature, pH and nitrite accumulation on the nitrification process. The ammonia removal efficiency during summer and winter and the nitrite accumulation in presence of free ammonia were observed. The biomass density was measured at different filter bed heights and the sludge production from the effluent of the backwashing water was evaluated. The results obtained were used to calibrate a mathematical model for the prediction of the ammonia removal profile in the filter bed and of biomass thickness.

Ammonia↗

PubMed2004

Pathogen removal efficiency from UASB + BF effluent using conventional and UV post-treatment systems.

The aim of this study was to verify the efficiency of removal of microorganisms in effluents of a Wastewater Treatment Plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The WWTP designed to treat domestic wastewater from a population of 1,000 inhabitants showed high removal efficiency for organic matter and suspended solids. Helminth eggs were also efficiently removed from the tertiary effluent and were found in the sludge from the UASB reactor; however, removal of bacteria in this system was very low. To enhance the efficiency of the system, the effluent from tertiary filters was submitted to UV disinfection in a real scale reactor. Our results showed that UV irradiation was very effective at lowering the concentrations of E. coli, thermotolerant coliforms and coliphages to acceptable levels for agricultural reuse. Salmonella spp. and helminth eggs were seeded into the tertiary effluent before passing through the UV reactor. Salmonella was not found in the final effluent, but helminth eggs were not completely inactivated by UV irradiation and viable eggs were detected after 28 d of incubation.

Agriculture↗

PubMed2004

All in the family: evidence for four new LEM-domain proteins Lem2 (NET-25), Lem3, Lem4 and Lem5 in the human genome.

LEM-domain proteins share a folded structure, the 'LEM-domain', which binds a conserved chromatin protein named BAF. Most LEM-domain proteins are found at the nuclear membrane, but some are nucleoplasmic. All characterized members of this family bind nuclear lamin filaments. We summarize the 'founding' LEM-domain proteins LAP2, emerin and MAN1 ('SANE' or 'XMAN' in Xenopus) and their emerging roles in gene regulation and nuclear assembly. These roles are placed in the context of human diseases ('laminopathies') caused by mutations in either emerin or A-type lamins. Other LEM-domain proteins might modify the phenotype or severity of human laminopathy, or cause new laminopathies. We summarize evidence that the human genome encodes at least four additional LEM-domain proteins, designated Lem2 (NET-25), Lem3, Lem4 and Lem5. Early adaptation of a consistent nomenclature, such as the "Lem" names proposed here, will facilitate rapid progress in this field. Further investigation of 'founder' and novel members of this family will be important to understand nuclear structure, and presents new opportunities to understand human disease.

Cell Nucleus↗