Computerized, comprehensive databases of cellular and secreted proteins from normal human embryonic lung MRC-5 fibroblasts: identification of transformation and/or proliferation sensitive proteins

Databases of protein information from human embryonal lung fibroblasts (MRC-5) have been established using computer analyzed two-dimensional gel electrophoresis. One thousand four hundred and eighty-two cellular proteins (1060 with isoelectric focusing and 422 with nonequilibrium pH gradient electrophoresis, in the first dimension) ranging in molecular mass between 8 and 234 kDa were separated and numbered. Information entered in the database (in most cases for major proteins) includes: protein name, HeLa protein catalog number, mouse protein catalog number, proteins matched in transformed human epithelial amnion cells (AMA) and peripheral blood mononuclear cells (PBMC), transformation and/or proliferation sensitive proteins, synthesis in quiescent cells, cell cycle regulated proteins, mitochondrial and heat shock proteins, cytoskeletal proteins and proteins whose synthesis is affected by interferons. Additional information entered for a few transformation-sensitive proteins that have been selected for future studies includes levels of synthesis and amounts in fetal human tissues. A total of four hundred and seventy-six [35S]methionine labeled polypeptides (258 isoelectric focusing; 218, nonequilibrium pH gradient electrophoresis) secreted by MRC-5 fibroblasts were separated and recorded (J. E. Celis et al., Leukemia 1987, 1, 707-717). Information entered in this database includes molecular weight and transformation sensitive proteins. These databases, as well as those of epithelial and lymphoid cell proteins (J. E. Celis et al., Leukemia 1988, 9, 561-601), represent the initial stages of a systematic effort to establish comprehensive databases of human protein information. In the long run, these databases are expected to offer a useful framework in which to focus the human genome sequencing effort.     

Analysis of trace metals in water by in-situ sample pre-concentration combined with wavelength dispersive X-ray fluorescence spectroscopy and inductively coupled plasma-optical emission spectroscopy

In this paper an earlier work on the use of poly(acrylamidoxime) cloth for use as a passive monitor for trace metals in water by characterizing the sorption of several metals onto the cloth using a continuous flow chamber is extended. The monitors consists of amidoxime chelating groups covalently bound to the surface of a textile encased in a common 35 mm slide holder. Placement of this device in the water to be sampled resulted in the uptake of heavy metals by the chelating groups. After removal of the monitor from water, the metals were analyzed using wavelength dispersive X-ray fluorescence spectroscopy (WDXRF) as well as by acid extraction followed by inductively coupled plasma-optical emission spectroscopy (ICP-OES). Correlation between the two methods of analysis varied with maximum correlation of 0.99897 observed for Pb and a minimum correlation of 0.16512 for Mg. The order of the distribution coefficients for the seven metals tested was: Pb> or =Cu>Zn>Cd>Mn>Fe>Mg, in agreement with the order of the stability constants for the amidoxime/hydroxamic acid group for the chelation of the same metals at a pH of 5, with the exception of Fe. Field testing of the monitors was also carried out and a comparison made between active sampling of river water and sampling with the monitors. Results indicated that semiquantitative analysis of trace metals in water may be performed using the passive monitors.     

Microextraction of volatile organic compounds using the inside needle capillary adsorption trap (INCAT) device

A novel method of solventless extraction has been developed based on a combination of solid phase micro extraction and purge and trap methods. In this technique, a hollow needle with either a short length of GC capillary column placed inside it, or an internal coating of carbon, is used as the preconcentration device. Sampling may be performed on ambient air, on solution, or the solution headspace, by passing the gas or liquid through the device either actively with a syringe, or passively via diffusion. The VOC are sorbed and concentrated onto either the carbon layer, or the liquid stationary phase of the capillary column, within the needle. Placing the needle into a heated GC injection port thermally desorbs the organic compounds directly into the GC without the need for solvent extraction. Results suggest that this procedure provides a rapid and sensitive alternative method to those currently available.     

Comprehensive two-dimensional gel protein databases offer a global approach to the analysis of human cells: the transformed amnion cells (AMA) master database and its link to genome DNA sequence data

A total of 3430 polypeptides (2592 cellular; 838 secreted) from transformed human amnion cells (AMA) labeled with [35S]methionine were separated and recorded using computer-aided two-dimensional (2-D) gel electrophoresis. A master 2-D gel database of cellular protein information that includes both qualitative and quantitative annotations has been established. The protein numbers in this database differ from those reported in an earlier version (Celis et al. Leukemia 1988, 2,561-602) as a result of changes in the scanning hardware. The reported information includes: percentage of total radioactivity recovered from the gels (based on quantitations of polypeptides labeled with a mixture of 16 14C-amino acids), protein name (including credit to investigators that aided identification), antibody against protein, cellular localization, (nuclear, 40S hnRNP, 20S snRNP U5, proteasomes, endoplasmic reticulum, mitochondria, Golgi, ribosomes, intermediate filaments, microfilaments and microtubules), levels in fetal human tissues, partial protein sequences (containing information on 48 human proteins microsequenced so far), cell cycle-regulated proteins, proteins sensitive to interferons alpha, beta, and gamma, heat shock proteins, annexins and phosphorylated proteins. The results presented should be considered as the initial phase of a joint effort between our laboratories to undertake a general and systematic analysis of human proteins. Using this integrated approach it will be possible to identify phenotype-specific proteins, to microsequence them and store the information in the database, to identify the corresponding genes, to search for homology with previously characterized proteins and to study the function of groups of proteins (pathways, organelles, etc.) that exhibit interesting regulatory properties. In particular, the 2-D gel protein database may become increasingly important in view of the concerted effort to map and sequence the entire human genome.     

Chelating polymers and related supports for separation and preconcentration of trace metals

This review is concerned mainly with the applications of chelating polymeric resins for the separation and concentration of trace metals from oceans, rivers, streams and other natural systems. Commercially available resins, specially prepared polymers and a selection of other sorbents are described and their uses outlined. Special emphasis is placed on the preconcentration of uranium from sea-water.     

The master two-dimensional gel database of human AMA cell proteins: towards linking protein and genome sequence and mapping information (update 1991).

The master two-dimensional gel database of human AMA cells currently lists 3801 cellular and secreted proteins, of which 371 cellular polypeptides (306 IEF; 65 NEPHGE) were added to the master images during the last 10 months. These include: (i) very basic and acidic proteins that do not focus under normal running conditions and (ii) low-abundant proteins that can only be detected after prolonged gel exposure. Annotation categories updated in this version include "protein name", "antibody against protein", "cellular localization", and "microsequenced proteins". New entries include "human autoantigens" and "cDNAs". For convenience we have included an alphabetical list of all known proteins recorded in this database. In the long run, the main goal of this database is to link protein and DNA sequencing and mapping information (Human Genome Program) and to provide an integrated picture of the expression levels and properties of the thousands of proteins that orchestrate various cellular functions both under physiological and abnormal conditions.     

Extraction of uranyl nitrate from aqueous nitrate solutions by open cell polyurethane foam sponge (OCPUFS)

The extraction of uranyl nitrate into open cell polyurethane foam sponge (OCPUFS) from aqueous solution, in the presence of salting agents, has been examined. The extraction efficiency was observed to depend on the concentration of uranyl and nitrate ions. The charge of the cation was also found to influence the distribution ratio. The effect of the change in temperature and pH was also studied. The results are interpreted in terms of OCPUFS acting as a viscous organic ether of moderate dielectric constant.     

A comprehensive two-dimensional gel protein database of noncultured unfractionated normal human epidermal keratinocytes: towards an integrated approach to the study of cell proliferation, differentiation and skin diseases.

A two-dimensional (2-D) gel database of cellular proteins from noncultured, unfractionated normal human epidermal keratinocytes has been established. A total of 2651 [35S]methionine-labeled cellular proteins (1868 isoelectric focusing, 783 nonequilibrium pH gradient electrophoresis) were resolved and recorded using computer-aided 2-D gel electrophoresis. The protein numbers in this database differ from those reported in an earlier version due to changes in the scanning hardware (Celis et al., Electrophoresis 1990, 11, 242-254). Annotation categories reported include: "protein name" (listing 207 known proteins in alphabetical order), "basal cell markers", "differentiation markers", "proteins highly up-regulated in psoriatic skin", "microsequenced proteins" and "human autoantigens". For reference, we have also included 2-D gel (isoelectric focusing) patterns of cultured normal and psoriatic keratinocytes, melanocytes, fibroblasts, dermal microvascular endothelial cells, peripheral blood mononuclear cells and sweat duct cells. The keratinocyte 2-D gel protein database will be updated yearly in the November issue of Electrophoresis.     

Light paraffin oxidative conversion in a silent electric discharge

Methane, ethane, and their mixtures with oxygen have been passed through the silent electric discharge at various operating conditions Beside.carbon oxides and water, formaldehyde and methane, and acetaldehyde and ethanol from ethane were the other major products. The effect of temperature, pressure, discharge voltage, contact time, and oxygen concentration have been examined. The rate of oxygen consumption is almost independent of oxygen concentration and depends mainly on the discharge voltage. The selectivities of methanol formation from methane and ethanol/formation from ethane were 20 and 15%, respectively.     

The CO2 Reforming of Natural Gas in a Silent Discharge Reactor

The conversion of a 1:1 CH ₄ /CO ₂ mixture to H ₂ and CO was shown to occur at ambient temperature in a silent discharge reactor. Conversions of up to 50% were achieved with two reactors in series with a total residence time of 3 min at 13kV and 25 mA.