PubMed Health⌕ Search

Biomedical subjects

W Whitfield

Publications and source records attributed to W Whitfield.

At least 19 recordsLinked to original sources
PubMed2005

The centrosomal protein CP190 regulates myosin function during early Drosophila development.

Centrosomes are the main microtubule (MT)-organizing centers in animal cells, but they also influence the actin/myosin cytoskeleton. The Drosophila CP190 protein is nuclear in interphase, interacts with centrosomes during mitosis, and binds to MTs directly in vitro. CP190 has an essential function in the nucleus as a chromatin insulator, but centrosomes and MTs appear unperturbed in Cp190 mutants. Thus, the centrosomal function of CP190, if any, is unclear. Here, we examine the function of CP190 in Cp190 mutant germline clone embryos. Mitosis is not perturbed in these embryos, but they fail in axial expansion, an actin/myosin-dependent process that distributes the nuclei along the anterior-to-posterior axis of the embryo. Myosin organization is disrupted in these embryos, but actin appears unaffected. Moreover, a constitutively activated form of the myosin regulatory light chain can rescue the axial expansion defect in mutant embryos, suggesting that CP190 acts upstream of myosin activation. A CP190 mutant that cannot bind to MTs or centrosomes can rescue the lethality associated with Cp190 mutations, presumably because it retains its nuclear functions, but it cannot rescue the defects in myosin organization in embryos. Thus, CP190 has distinct nuclear and centrosomal functions, and it provides a crucial link between the centrosome/MT and actin/myosin cytoskeletal systems in early embryos.

Animals↗
PubMed1998

Titration of a CD45-FITC conjugate to determine the linearity and dynamic range of fluorescence intensity measurements on lymphocytes.

To produce biologic calibrators for relative fluorescence intensity (RFI) measurements, we stained leukocytes with serial dilutions of CD45-FITC conjugate and processed them using our regular whole blood lysis procedure. Cells were stained with conjugate concentrations ranging from twice recommended to a million-fold lower. At the highest concentrations of conjugate, the RFI reached a plateau near the top of the third decade, indicating saturation of CD45 binding sites. As the concentration decreased, the RFI declined in a highly linear relationship between the dilution factor and the histogram channel number. For channel numbers corresponding to the lowest percentiles of the RFI distribution, linearity persisted down to the first half decade. The slope of this relationship revealed a true dynamic range of 4.5 decades, which was comparable to the value obtained with microbead standards calibrated in molecules of equivalent soluble fluorochrome (MESF). Our results suggest that the lower limit of linearity for fluorescence intensity from fluorescein isothiocyanate (FITC)-stained lymphocytes is below 500 MESF and that cellular autofluorescence is the major limiting factor in detecting and quantifying FITC-specific staining. This procedure provides an adroit way of characterizing the linearity and dynamic range of measurements for quantitative fluorescence cytometry using exactly the same matrix, stains, and preparation methods as those used for cellular analytes.

Antibodies, Monoclonal↗
PubMed1990

Psychosocial rehabilitation: the role of the psychiatric nurse.

The roles and functions of Nurses working in Rehabilitation Psychiatry are varied and complex, new challenges are arising out of historical and emerging contexts. This paper explores problems faced by the Psychiatric Nurse and explains some of the functions and roles that contribute to the rehabilitation process.

Humans↗
PubMed1987

Quantitative differences among various proteins as blocking agents for ELISA microtiter plates.

We tested instantized dry milk, casein, gelatins from pig and fish skin, serum albumin and several other proteins for their abilities to block non-specific binding (NSB) of a peroxidase-conjugated immunoglobulin to polystyrene microtiter plate wells. Each blocking protein was tested across a million-fold concentration range, both in simultaneous incubation with the peroxidase conjugate and as a pretreatment agent where excess protein was washed away before incubation with the conjugate. Overall, instantized milk and casein were the most effective proteins tested: they inhibited NSB by over 90% in both the simultaneous and pretreatment modes at far lower concentrations than most of eight other proteins. Enzymatically hydrolyzed porcine skin gelatin was the least effective protein tested: it did not reduce NSB by more than 90% even at its highest concentrations; its blocking ability fell rapidly upon dilution; and it was almost useless as a pretreatment agent. Fish skin gelatin showed much better blocking activity than hydrolyzed porcine gelatin, and it still had the practical advantage of remaining fluid even under refrigeration. Our results suggest that some proteins (such as casein) block NSB to plastic primarily through protein-plastic interactions, while others (such as porcine skin gelatin) block primarily through protein-protein interactions. Although the optimal blocking agent for any particular ELISA system must be determined by empirical testing, these results should be helpful in selecting the best possible candidate proteins for further evaluation.

Animals↗
PubMed1982

Liquid-chromatographic measurement of phenylalanine and tyrosine in serum.

With phenylalanine ammonia-lyase (EC 4.3.1.5) we converted phenylalanine (Phe) and tyrosine (Tyr) to transcinnamic acid and p-coumaric acid, respectively. These were separated by "high-performance" liquid chromatography and detected at 280 nm. We measured the Phe and Tyr content of human serum by adding 100 mU of the enzyme to a 20-microL serum aliquot, mixing for 2 h at 24 degrees C, then stopping the reaction with 1 mL of cold methanol. Precipitated proteins were removed by centrifugation and the separated clear supernates were stored at -20 degrees C. For chromatographic separation, detection, and quantification, we used a system equipped with a C-18 reversed-phase column, a variable-wavelength spectrophotometer, a printer-plotter, and a microcomputer. The mobile phase was a mixture of dilute aqueous (50 g/L) acetic acid and CH3CN (80/20, by vol). CVs for specimens containing 100 mg of Phe or Tyr per liter varied from 5 to 10%. Analytical recoveries were near 100%.

Centrifugation↗
PubMed1981

Analysis of reconstruction of an RNP particle which stores 5S RNA and tRNA in amphibian oocytes.

Previtellogenic oocytes of Triturus cristatus accumulate a free cytoplasmic RNP which sediments at 40S and contain 5S RNA and tRNA in association with two proteins of MW 45,000 and 39,000 daltons (P45 and P39). The 40S particle has a buoyant density of 1.53 g . cm-3 in CsCl and consists of four identical RNP subunits. Each monomeric subunit contains one molecule of 5S RNA, three molecules of tRNA, two molecules of P45 and one molecule of P39. The 40S particle can be completely dissociated by SDS treatment into its individual components, and the subunits, and even the complete 40S particle, can be reformed by removal of SDS in the presence of 0.2 M NaCl. RNA/protein binding experiments with isolated components, and analysis of reformed RNP complexes in CsCl gradients, demonstrate that the stable interactions are: 5S RNA/P45, 3(tRNA)/P45, 5S RNA/P39 and 5S RNA/P45/P39. Immunological studies show that P45 has also a nuclear location and may bind to the 5S RNA transcript in the chromatin, whereas P39 is predominantly cytoplasmic and is possibly related to proteins associated with 5S RNA in the ribosomal 60S subunit. It is suggested that the 40S RNP particle not only stores 5S RNA and tRNA but also provides a means for the exchange of the 5S RNA transcript binding protein (P45) for the 5S RNA ribosome associated protein (P39).

Animals↗
PubMed1977

The epilepsies.

Explore the source record for details and available documents.

Epilepsies, Partial↗
PubMedDate not supplied

Development and evaluation of a microtiter plate enzyme immunoassay for antibodies to 3,3'-dichlorobenzidine.

The authors obtained and evaluated antisera from rabbits injected with a derivative of a potent bladder carcinogen, dichlorobenzidine (DCB), conjugated to bovine serum albumin (BSA). A 14C-radioimmunoassay (RIA) was able to detect the presence of DCB antibodies, but its relative insensitivity led to the development of a more sensitive enzyme immunoassay (EIA). The EIA test was a "sandwich" method in which a second antibody, labeled with an enzyme (horseradish peroxidase), was used to measure antibody binding to transferrin (Tf)-conjugated DCB immobilized on a microtiter plate. Antibody titers measured by RIA were approximately 1:40; when measured by EIA, they were approximately 1:40,000. Antibody specificity was assessed by comparing the antibody binding activities of DCB, BSA, Tf, BSA-conjugated to DCB, and a number of N-substituted aromatic compounds that included benzidine (Bz). Among the compounds tested, the rabbit antiserum reacted only with DCB and the carrier protein, BSA. Moreover, antibody binding activity to Tf-conjugated DCB was significantly inhibited by unconjugated DCB concentrations between 30 and 500 ng/mL. The precision of antibody binding activities as a function of DCB concentration (expressed by the CV) ranged from 9% for low (30 ng/mL) DCB levels to 12% for higher (500 ng/mL) levels. This evaluation suggests that the antiserum obtained would be appropriate for detecting DCB levels at the ng/mL level.

3,3'-Dichlorobenzidine↗