Production and characterization of two monoclonal antibodies against human myeloperoxidase

Two monoclonal antibodies against human myeloperoxidase, designated 3-2H3 and 4-2C11, were produced and characterized. Both bound to the native enzyme, but neither bound to the denatured enzyme or to its two denatured subunits. 4-2C11 bound to the three types of leukocyte myeloperoxidase, I, II, and III, as well as to the four types of myeloid leukemia HL-60 cell myeloperoxidase, IA, IB, II, and III. 3-2H3 did not bind to enzyme IB, but bound to the other types of leukocyte and HL-60 cell enzymes. On incubation with myeloperoxidase III, 4-2C11 inhibited the enzyme activity, but 3-2H3 did not. Both antibodies belong to the IgG1 subclass.     

Bluejay 1.0: genome browsing and comparison with rich customization provision and dynamic resource linking

Background  

The Bluejay genome browser has been developed over several years to address the challenges posed by the ever increasing number of data types as well as the increasing volume of data in genome research. Beginning with a browser capable of rendering views of XML-based genomic information and providing scalable vector graphics output, we have now completed version 1.0 of the system with many additional features. Our development efforts were guided by our observation that biologists who use both gene expression profiling and comparative genomics gain functional insights above and beyond those provided by traditional per-gene analyses.     

The combination of berberine and irradiation enhances anti‐cancer effects via activation of p38 MAPK pathway and ROS generation in human hepatoma cells

Berberine, an isoquinoline plant alkaloid, has been known to generate a wide variety of biochemical and pharmacological effects. In order to elucidate the molecular mechanism for the berberine‐induced enhancement of radio‐sensitization, the human hepatoma HepG2 cells were treated with berberine combined with irradiation. The anti‐tumor effect of gamma radiation was found to be significantly enhanced by berberine. The evidences of apoptosis, such as apoptotic DNA fragmentation and annexin V staining, were observed in the cells treated with the combination of berberine and irradiation. Additionally, the levels of reactive oxygen species (ROS) and nitric oxide (NO) were apparently elevated in the combination system. The activations of p38, Bax, and caspase‐3 were also detected in the irradiated cells pretreated with berberine. The productions of ROS and annexin V staining in the cells treated with the combination of berberine and irradiation were significantly inhibited by the specific inhibitor of p38 MAPK, SB203580. The cell death induced by berberine alone or the combination of berberine and irradiation was suppressed by the anti‐oxidant, N‐acetyl cysteine (NAC). Taken together, the present results clearly indicate that the combination of berberine and gamma‐radiation enhance the anti‐cancer effects through the p38 MAPK pathway and ROS generation. J. Cell. Biochem. 107: 955–964, 2009. © 2009 Wiley‐Liss, Inc.     

Embryonic Stem Cell-Derived Extracellular Vesicles Maintain ESC Stemness by Activating FAK

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Regional distribution of the enzymes and substrates mediating the action of cAMP in the mammalian lens

Localization of adenylate cyclase activity in the outer cortical regions of the bovine lens correlates with the restriction of the Gs and Gi guanine nucleotide regulatory subunits of this enzyme to these same regions of the lens. In contrast, the major membrane substrates for cAMP-dependent protein kinase (cAMP-PK) (molecular masses of 18, 26 and 28 kDa) were identified in both the inner nuclear and the outer cortical regions of the lens. However, there were differences in the relative amounts of Pi incorporated into the 18 kDa and 28 kDa components in different lens regions. The three major membrane substrates for cAMP-PK were also phosphorylated when homogenates of lens cortex were incubated with [gamma-32P]ATP plus activators of the lens adenylate cyclase. In contrast, there was no incorporation of 32P into these substrates when homogenates of lens nucleus were used. When exogenous cAMP was added to homogenates of lens nucleus or cortex, 32P was incorporated into the membrane substrates for cAMP-PK in both regions of the lens, indicating that cAMP-PK was present in both regions. Interestingly, cAMP phosphodiesterase activity was at least 10-times greater in lens cortex than in the lens nucleus. These results indicate that while the major membrane substrates for cAMP-PK could be phosphorylated in all regions of the lens, there is a restriction of those enzymes that synthesize and degrade cAMP to the outer cortical regions of this organ.     

An artificial neural network for biomass estimation from automatic pH control signal

This study developed an artificial neural network (ANN) to estimate the growth of microorganisms during a fermentation process. The ANN relies solely on the cumulative consumption of alkali and the buffer capacity, which were measured on-line from the on/off control signal and pH values through automatic pH control. The two input variables were monitored on-line from a series of different batch cultivations and used to train the ANN to estimate biomass. The ANN was refined by optimizing the network structure and by adopting various algorithms for its training. The software estimator successfully generated growth profiles that showed good agreement with the measured biomass of separate batch cultures carried out between at 25 and 35_C.     

Integration of working environment into life cycle assessment framework

Background, aim, and scope  Life cycle assessment (LCA) has been considered one of the tools for supporting decision-making related to the environmental aspects of a product system. It has mainly been used to evaluate the potential impacts associated with relevant inputs and outputs to/from a given product system throughout its life cycle. In most cases, LCA has not considered the impacts on the internal environment, i.e. working environment, but only the external environment. Recently, it has been recognized that the consideration of the impacts on the working environment as well as on the external environment, is needed in order to assess all aspects of the effects on human well-being. To this end, this study has developed a total environmental assessment methodology which enables one to integrate both the working environment and the external environment into the conventional LCA framework. Materials and methods  In general, the characteristics of the impacts on the external environment are different from those on the working environment. In order to properly integrate the two types into total environmental impacts, it is necessary to define identical system boundaries and select impact category indicators at the same level. In order to define the identical system boundary and reduce the uncertainties of LCI results, the hybrid IOA (input–output analysis) method, which integrates the advantages between conventional LCI method and IOA method, is introduced to collect input and output data throughout the entire life cycle of a given product. For the impact category indicators at the endpoint level, LWD (Lost Work Days) is employed to evaluate the damage to human health and safety in the working environment, while DALY (disability-adjusted life years) and PAF (Potentially Affected Fraction) are selected to evaluate the damage to human health and eco-system quality in the external environment, respectively. Results and discussion  The environmental intervention factors (EIFs) are developed not only for the data categories of resource use, air emissions, and water emissions, but also for occupational health and safety to complete a life cycle inventory table. For the development of the EIFs on occupational health and safety, in particular, the number of workers affected by i hazardous items and the number of workers affected at the i magnitude of disability are collected. For the characterization of the impact categories in the working environment, such as occupational health and safety, the exposure factors, effect factors, and damage factors are developed to calculate the LWD of each category. For normalization, the normalization reference is defined as the total LWD divided by the total number of workers. A case study is presented to illustrate the applicability of the proposed method for the integration of the working environment into the conventional LCA framework. Conclusions  This study is intended to develop a methodology which enables one to integrate the working environmental module into the conventional LCA framework. The hybrid IOA method is utilized to extend the system boundary of both the working environment module and the external environment module to the entire life cycle of a product system. In this study, characterization models and category indicators for occupational health and safety are proposed, respectively, while the methodology of Eco-indicator 99 is used for the external environment. In addition to aid further understanding on the results of this method, this study introduced and developed the category indicators such as DALY, and LWD, which can be expressed as a function of time, and introduced PAF, which can be expressed as a probability. Recommendations and perspectives  The consideration of the impacts not only on the external environment, but also on the working environment, is very important, because the best solution for the external environment may not necessarily be the best solution for the working environment. It is expected that the integration of occupational health and safety matters into the conventional LCA framework can bring many benefits to individuals, as well as industrial companies, by avoiding duplicated measures and false optimization.