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Biomedical subjects

J C Boyd

Publications and source records attributed to J C Boyd.

At least 19 recordsLinked to original sources

Estrogen inhibits calbindin-D28k expression in mouse uterus.

The cellular localization and hormonal controls of calbindin-D9k expression in the rodent reproductive tract have suggested new functions for this protein. The present studies were undertaken to extend the earlier studies of calbindin-D9k to the related protein, calbindin-D28k. Immunohistochemical studies revealed that calbindin-D28k was absent from female rat reproductive tissues, but was abundantly expressed in immature mouse uterus and oviduct. Immunoreactivity was restricted to the endometrial and glandular epithelium of the uterus and the oviductal epithelium. Neither 1,25-dihydroxyvitamin D- nor strontium-containing diets (to blunt 1,25-dihydroxyvitamin D production) affected expression of calbindin-D28k. Uterine, but not oviductal, calbindin-D28k decreased markedly at sexual maturity; this pattern persisted in pregnant mice and was reproduced in immature mice by the administration of estradiol (3 micrograms/day for 3 days). RNA extraction and Northern analyses demonstrated that estrogen markedly decreased calbindin-D28k mRNA abundance in the uterus, but not in the oviduct. These findings suggest that estrogen affects mammalian calbindin-D28k expression and represent a rare example of estrogen-induced down-regulation of gene expression.

Animals

Characterization of the variation between batches of Fast-Flo lactose using low frequency dielectric spectroscopy.

The dielectric response of four batches of lactose has been measured over a frequency range of 10(4) to 10(-2) Hz. The spectra corresponding to three of the batches were identical, while the fourth showed a marked reduction in response. This particular batch has also been reported to exhibit longer disintegration times than the other three when formulated as a tablet. The potential use of dielectric spectroscopy as a means of screening batches of pharmaceutical materials is discussed.

Chemistry, Pharmaceutical

Relationship of invasiveness to proliferating activity and to cytoskeletal protein production in human neuroepithelial tumors maintained in an organ culture system: use of human cortex and dura as supporting matrices.

The proliferation and invasiveness of cultured human neuroepithelial tumors were studied. A human malignant astrocytic glioma cell line (U-251 MG) and a medulloblastoma cell line (D283 Med) were maintained for 3 weeks in an organ culture system using adult human brain cortex, dura mater, or Gelfoam sponge as growth matrices. The cells were labeled with bromodeoxyuridine (BrdU) at different time points, and immunohistochemistry was performed for BrdU, glial fibrillary acidic (GFA) protein (in U-251 MG), and neurofilament (NF) protein (in D283 Med). In the U-251 MG line, the cells grew successfully in each matrix, forming a fibrillated solid area and a peripheral zone of invasion. The labeling index (LI) expressed as the percentage of BrdU-labeled cells and the percentage of GFA protein-positive cells in the two zones of the explants were analyzed. The LIs in all cultures were significantly higher in the peripheral than in the central zones. On the other hand, the percentage of GFA protein-positive cells in each matrix was greater in the central zone than in the periphery. The LI was inversely correlated with the percentage of GFA protein-positive cells over the areas counted in each growth matrix. GFA protein production in cells grown on cortex and on dura mater was significantly higher than that in cells grown on Gelfoam. In the D283 Med line, the cells formed an aggregated zone, with peripheral cells infiltrating the Gelfoam. This line showed poor growth on human cortex. Cells grown on the dura demonstrated an LI similar to that on Gelfoam, and cells often infiltrated the dura.(ABSTRACT TRUNCATED AT 250 WORDS)

Brain Neoplasms

Kinetics and glial fibrillary acidic (GFA) protein production in a transplantable human giant cell glioblastoma (D-212 MG) of near haploid karyotype maintained in an organ culture system. An immunohistochemistry study.

A transplantable subcutaneous tumour (designated D-212 MG), sequentially passaged in athymic nude mice and originally derived from a human giant cell glioblastoma, was maintained in an organ culture (matrix) system and studied immunohistochemically after in vitro pulse-labelling with bromodeoxyuridine (BrdU) and for the presence of glial fibrillary acidic (GFA) protein, after 1, 2 and 3 weeks in culture. The histological characteristics of the tumour, showing two cell populations of giant multinucleated cells and small cells, were preserved in the explants. An increased percentage of multinucleated giant cells was found after 3 weeks in vitro. A small but constant fraction (4-6%) of these cells continued to synthesize DNA. The labelling index of the small cells was somewhat higher, but decreased slightly although significantly over the 3-week period in vitro (from approximately 10.5 to 8%). The percentage of small cells that were positive for GFA protein was in the region of 75% and that of the giant multinucleated cells was in the region of 45%; it did not change significantly during the 3 weeks in vitro. The in vitro results confirm the astrocytic nature of both the small cells and the giant multinucleated cells in this tumour, the capacity of both cell populations to synthesize DNA in culture and to demonstrate invasiveness, and suggest the possibility that some of the giant multinucleated cells may have originated from the conversion of a number of small tumour cells.

Glial Fibrillary Acidic Protein

Robotics in the medical laboratory.

Robotic systems specifically designed for the automation of laboratory tasks are now available commercially. Equipped with computer, analytical hardware, and supporting software, these devices may soon revolutionize the concept of the clinical laboratory and usher in a new era in laboratory testing. We review the types of robots and motion-control software currently available and discuss examples of their applications that extend across many analytical areas. Several ongoing projects are concerned with the systematic integration of robotic devices with other laboratory automation. The integrated robotic laboratories emerging from this work portend a bright future for robotic automation. Many challenges remain, however, in training the individuals needed to develop and manage robotic laboratories, and in making this new technology cost-efficient.

Laboratories, Hospital

On dividing reference data into subgroups to produce separate reference ranges.

We consider statistical criteria for partitioning a reference database to obtain separate reference ranges for different subpopulations. Using general formulas relating population variances, sample sizes, and the normal deviate test for the significance of the difference between two subgroup means, we show that partitioning into separate ranges produces little reduction in between-person variability, even when the differences between means are highly significant statistically. However, when there is a clear physiological basis for distinguishing between certain subgroups, simulation studies show that partitioning may be necessary to obtain reference limits that cut off the desired proportions of low and high values in each subgroup. Guidelines based on these results are provided to help decide whether separate ranges should be obtained for a given analyte.

Analysis of Variance

Relationship of the demonstration of intermediate filament protein to kinetics of three human neuroepithelial tumor cell lines. Lack of neural-related proteins in most cells in S phase: a double-labeled immunohistochemical study on matrix cultures.

The immunocytochemical demonstration of intermediate filament proteins in three human neuroepithelial tumor cell lines maintained in vitro on a three-dimensional matrix was correlated with the proportion of cells in S phase. The cell lines of a medulloblastoma (D283 Med), a retinoblastoma (WERI-Rb1), and an astrocytic glioma (U-251 MG) were cultured in an organ culture system, pulse-fed with bromodeoxyuridine, and double-labeled by immunoperoxidase and by the avidin-biotin peroxidase complex method for bromodeoxyuridine and for intermediate filament proteins [each triplet of neurofilament proteins, as well as vimentin and glial fibrillary acidic (GFA) protein] using eight different antibodies. The average percentages of bromodeoxyuridine-labeled cells for the D283 Med, WERI-Rb1, and U-251 MG lines were respectively, 25, 32, and 12% 30 minutes after pulse labeling. In the D283 Med line, 15- greater than 95% of the cells were positive for each neurofilament protein, and 80% of the cells were positive for vimentin; less than 10% of the cells in S phase were positive with each of the five antineurofilament protein monoclonal antibodies (Mabs), but 20% of the vimentin-positive cells were in S phase. In the WERI-Rb1 line, 44 and greater than 96% of the cells were positive for the high-molecular-weight neurofilament subunit and high- and middle-molecular-weight neurofilament subunits proteins, respectively, but only 5% of the high-molecular-weight neurofilament positive cells were in S phase. In the U-251 MG line, 37 and 98% of the cells were positive for GFA protein and vimentin, respectively; only 3% of the GFA protein-positive cells, but 13% of the vimentin-positive cells, were in S phase. The results indicate that, when maintained in a matrix culture system, most cells in S phase in these lines lack markers of neural differentiation.

Astrocytoma

Comparison of discriminant analysis procedures in laboratory differentiation of hypercalcemia.

Logistic, linear, and quadratic discriminant analyses were compared in their ability to differentiate hypercalcemic patients with primary hyperparathyroidism from those with malignancy. Linear and quadratic discriminant analyses were performed by use of both untransformed and logarithmically transformed data. Application of principal components analysis with varimax rotation was helpful in revealing the underlying relationships between variables. All discriminant methods identified serum albumin as the best single discriminating test, with the log-quadratic discriminant analysis classifying 81% of patients correctly. The combination of albumin, carboxy-terminal parathyroid hormone, and chloride improved classification accuracy (92% by use of log-quadratic discriminant analysis). Logistic discriminant analysis, using all 20 variables, gave a classification accuracy of 100%. Quadratic discriminant analysis gave better classification than linear discriminant analysis, and both methods performed better when log-transformed data were used. Logistic discriminant analysis followed by discrimination procedures using log-transformed data yielded the highest classification accuracy and reliability of the methods used.

Humans

Ipratropium bromide treatment of experimental rhinovirus infection.

The importance of parasympathetic-cholinergic mechanisms in the production of common cold symptoms is not clear. The quaternary ammonium anticholinergic antagonist ipratropium bromide was intranasally administered under double-blind, randomized, placebo-controlled conditions to assess its tolerance and efficacy in reducing nasal hypersecretion in adult volunteers with experimental rhinovirus colds. Ipratropium was sprayed intranasally three times daily (80 micrograms per treatment) for 5 days beginning 24 h after intranasal inoculation of rhinovirus type 39. Clinical colds occurred in 50% of 30 infected ipratropium recipients and in 76% of 33 infected placebo recipients (P = 0.04). The nasal mucus weights tended to be lower for ipratropium-treated persons (mean +/- standard deviation, 14.7 +/- 15.1 g/5 days) than for placebo-treated recipients (24.7 +/- 28.0 g/5 days; P = 0.076). Whereas total nasal symptom scores were similar between the two groups, the rhinorrhea score analyzed for each day of treatment showed nonsignificant trends favoring the ipratropium group over the last 4 days of treatment. Ipratropium was generally well tolerated. The results suggest that cholinergic mechanisms are at least partially responsible for nasal mucus production in rhinovirus colds but that the effect of anticholinergic compounds alone is insufficient to be of practical use in treatment, although they may have value as components of multi-ingredient preparations.

Administration, Intranasal

Robotics in the clinical laboratory.

We are beginning to see the potential of robotics in the clinical laboratory through integration with automated analyzers and computer systems. However, there is a need for training programs that will prepare technologists to design and implement robotic systems for clinical laboratories. What will the robot laboratory of the future look like? We will see hospital laboratories begin to be located some distance away from the main facility because the labor component of staffing satellite laboratories will have been greatly reduced. Instrument manufacturers will see the need for analyzers that are robot-friendly and allow for simplified interfacing, both electronic and mechanical. Robots will become more versatile even to the point of performing complete instrument repair. Laboratories will be equipped with many task-oriented robotic stations, including, for example, accessioning and processing robots that prepare samples for transport by robotic carts. Analysis will be performed by a combination of robot and dedicated analyzer. Laboratory results will be reviewed by algorithms in the larger laboratory computer, which will alert the laboratory worker to unusual results. A large variety of analyses will be available to the patient with rapid turnaround. The end result will be more efficient health care delivery at reduced cost.

Computer Communication Networks

Activity measurements of calcium, sodium, potassium, and chloride after equilibrium dialysis used to show lack of evidence for protein interference with calcium electrodes.

We measured the activity of Ca2+, Na+, K+, and Cl- with ion-selective electrodes after equilibrium dialysis of solutions with different albumin concentrations. The calculated Donnan ratio was the same for all ions in the same solution and increased with the albumin concentration, as predicted by the Donnan theory. The Donnan distribution ratio for Ca2+ was similar, as determined with instruments from three different manufacturers. For healthy subjects and patients with renal stone disease, we did not find any correlation between serum concentrations of ionized calcium and albumin. The discordance between measured ionized calcium and albumin-corrected total calcium depended on the correction algorithm we utilized. The difficulties of absolutely proving or disproving a protein error in these measurements are discussed, but our data are not consistent with protein being a source of error in measurements of ionized calcium.

Adult

Methotrexate interferes with determinations of conjugated bilirubin with the Kodak Ektachem 400.

Immediately after intravenous infusion of a high dose (concentration in serum greater than 1000 mumol/L) of methotrexate, the apparent conjugated bilirubin (Bc) concentrations in serum of two osteosarcoma patients, as measured by the Kodak Ektachem 400 analyzer, were greater than the corresponding total bilirubin concentrations, but decreased as the concentrations of methotrexate in serum decreased. In an interference study we found that methotrexate added to sera containing a wide range of basal Bc concentrations increased the measured Bc concentration in a linear and dose-related fashion. Methotrexate also interfered negatively with measurements of unconjugated bilirubin (Bu). The source of the interference appears to be an overlap in the absorption spectrum of methotrexate with Bc and Bu at 400 nm.

Autoanalysis

Perspectives on the use of chemometrics in laboratory medicine.

Today's automated laboratory instruments are capable of generating prodigious volumes of high-quality measurements. Increasingly, the powerful mathematical and statistical methods of chemometrics are being called upon to help reduce these measurements to useful information. Chemometric methods have been important in automating various data-intensive functions of the clinical laboratory, including analysis of cellular images, identification of bacteria and fungi on the basis of their metabolic and chemical properties, and identification of drugs and toxic substances from their mass spectra. These methods also appear promising in aiding both the selection and interpretation of laboratory tests for diagnosis, monitoring, and prognosis. In spite of the demonstrated potential of these methods, significant problems remain to be solved in the areas of measurement standardization, data-base collection, and user familiarity with these approaches before chemometric methods can be used most fully by the clinical laboratory.

Artificial Intelligence