Immunoadsorbent purification of carcinoembryonic antigen using a monoclonal antibody: a direct comparison with a conventional method

Carcinoembryonic antigen (CEA) has been extracted and purified by two different methods from a single aqueous homogenate of liver metastases from a primary adenocarcinoma of the colon, thus enabling direct comparison to be made between a conventional technique using gel filtration and a monoclonal antibody immunoadsorbent method. Yield, immunoreactivity and purity have been determined in both cases by direct weighing, enzyme-linked immunoadsorbent assay, radioimmunoassay and SDS-polyacrylamide gel electrophoresis. Reactivity of the antigen with monoclonal and polyclonal anti-CEA antibodies recognising different epitopes was analysed by Western blotting. There appears to be no significant difference in immunoreactivity or purity by these criteria, but the immunoadsorbent method gave a higher yield of CEA for far less expenditure of time and effort. A variant of CEA with a lower molecular weight was also identified in both preparations.     

Buffered lidocaine as a local anesthetic: an investigation of shelf life.

STUDY OBJECTIVE: To determine whether buffered lidocaine must be prepared just before use. DESIGN: Randomized, double-blind, prospective trial. SETTING: University hospital. PARTICIPANTS: Twenty-four adult volunteers. INTERVENTIONS: Three buffered lidocaine solutions prepared seven days, one day, and just before use were compared with a control solution. Subjects received 0.5 mL intradermal injections of each solution. Pain of infiltration and extent and duration of anesthesia were measured. MEASUREMENTS AND MAIN RESULTS: Pain of infiltration was less with all buffered solutions than control (P less than .0001). Mean maximum diameter of anesthesia ranged from 29 to 33 mm for the buffered solutions compared with 31 mm for control. Mean duration of anesthesia was 33 minutes for control and 30 minutes for all of the buffered solutions. There was no statistically significant difference in extent or duration of anesthesia for any of the solutions (P greater than .5, beta = .15 for delta = 10%). CONCLUSION: Buffered lidocaine stays effective for up to one week after preparation. It is therefore convenient to use in emergency settings.     

Construction and validation of a Parkinson's disease mutation genotyping array for the Parkin gene.

Parkin mutations account for the majority of familial and sporadic early onset Parkinson's disease (EOPD) cases with a known genetic association. More than 100 mutations have been described in the Parkin gene that includes homozygous, compound heterozygous, and single heterozygous mutations. We have designed a Parkin mutation genotyping array (gene chip) that includes published Parkin sequence variants and allows their simultaneous detection. The chip was validated by screening 85 PD cases and 47 controls previously tested for Parkin mutations. Similar genotyping microarrays have been developed for other genetically heterogeneous diseases including age-related macular degeneration. Here, we show the utility of a genotyping array for Parkinson's disease by analysis of 60 subjects from the Genetic Epidemiology of Parkinson Disease (GEPD) study that includes 15 early-onset PD case probands and 45 relatives.     

Needs assessment for persons with severe mental illness: What services are needed for successful community living?

The development and implementation of effective community support systems are goals of many public mental health authorities who are attempting to shift the focus and dollars for mental health services from inpatient to community care. This article presents the results of a survey which asked 90 community mental health agency case managers to assess the community support and residential needs of over 1400 of their clients. Medication monitoring and therapy were rated high priority needs. Psychosocial treatment, day and vocational activities also ranked high. Survey responses regarding residential services indicated a need for more supported and supervised options.The study reported was a collaborative effort by county and agency staff.     

Effects of MR imaging on murine natural killer cell cytotoxicity

To determine the effect of MR imaging on the immune system, 21 male C57BL/6 X DBA/2 F1 mice were exposed to MR imaging at a field strength of 0.15 T for 2 hr. Another nine mice (controls) were sham exposed for the same amount of time. Mice were sacrificed and their spleens removed 24, 72, and 144 hr after the exposure (MR or sham). Spleen cell suspensions were passed over nylon wool columns and then used as effector cells in a short-term natural killer cell cytotoxicity assay with 51Cr-labeled YAC-1 cells as target cells. The results showed no evidence of decreased cytotoxicity due to exposure to MR. On the contrary, at all three times after exposure and for all target-to-effector cell ratios, mean cytotoxicity was greater for MR-exposed groups than for sham-exposed groups. The results show that MR exposure has no adverse effect on the immune system, as evidenced by natural killer cell activity.     

Cypress (Cupressus sempervirens) pollen allergens: identification by protein blotting and improved detection of specific IgE antibodies

On the basis of results of an investigation of the effects of different treatments employed, a dialysed and reduced extract of Cupressus sempervirens was separated electrophoretically on sodium dodecylsulphate-polyacrylamide gels before being transferred and then fixed with glutaraldehyde to nitrocellulose membrane. Probing with sera from 91 subjects allergic to C. sempervirens pollen followed by detection of bound IgE antibodies with [125I]-labelled anti-human IgE revealed 17 IgE-binding proteins in the molecular weight range 14-96 kilodaltons (kDa). One component, of molecular weight approximately 42 kDa, reacted with IgE antibodies in the sera of 81.3% of the allergic subjects and, for each of the subjects, this component bound the greatest quantity of IgE. Almost 50% of the sera recognized only the approximately 42 kDa component, reinforcing the conclusion that this component is the major allergen of C. sempervirens pollen. A comparative study employing C. sempervirens pollen allergen discs prepared commercially or in the laboratory showed that values of the uptakes of [125I]-anti-IgE indicating the presence of pollen-reactive IgE antibodies obtained with the latter discs were consistently higher (means 4.5 vs. 0.88), and that false-negative results were obtained when many sera were used with the commercial discs. The results of this study provide an essential basis for the production of standardized, safe and effective C. sempervirens pollen extract applicable to diagnosis and therapy of cypress pollen allergy.     

Normative data of hemoglobin concentration and free erythrocyte protoporphyrin in a private pediatric practice: a 1990 update

Free erythrocyte protoporphyrin (FEP) and hemoglobin (Hgb) concentrations were tested in 790 children in a private pediatric office; results were compared to those obtained in 1984. Only 16 children (2%) had abnormal FEPs in 1990 compared to 76 children (9.6%) in the earlier study. The mean FEP in the normal group also decreased significantly in each age group studied. The hemoglobin concentrations were not significantly different in most of the age groups studied. Screening for iron deficiency in our pediatric practice by determining hemoglobin and FEP concentrations had a much lower yield in 1990 than in 1984.     

Large-scale identification of mammalian proteins localized to nuclear sub-compartments

Many nuclear components participating in related pathways appear concentrated in specific areas of the mammalian nucleus. The importance of this organization is attested to by the dysfunction that correlates with mis-localization of nuclear proteins in human disease and cancer. Determining the sub-nuclear localization of proteins is therefore important for understanding genome regulation and function, and it also provides clues to function for novel proteins. However, the complexity of proteins in the mammalian nucleus is too large to tackle this on a protein by protein basis. Large-scale approaches to determining protein function and sub-cellular localization are required. We have used a visual gene trap screen to identify more than 100 proteins, many of which are normal, located within compartments of the mouse nucleus. The most common discrete localizations detected are at the nucleolus and the splicing speckles and on chromosomes. Proteins at the nuclear periphery, or in other nuclear foci, have also been identified. Several of the proteins have been implicated in human disease or cancer, e.g. ATRX, HMGI-C, NBS1 and EWS, and the gene-trapped proteins provide a route into further understanding their function. We find that sequence motifs are often shared amongst proteins co-localized within the same sub-nuclear compartment. Conversely, some generally abundant motifs are lacking from the proteins concentrated in specific areas of the nucleus. This suggests that we may be able to predict sub-nuclear localization for proteins in databases based on their sequence.