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Biomedical subjects

C M Huang

Publications and source records attributed to C M Huang.

At least 91 records · Page 5Linked to original sources

An enzymatic method for 5-fluorocytosine.

We developed an enzymatic method for determination of 5-fluorocytosine in serum, using creatine iminohydrolase (EC 3.5.4.21), the Cobas-Bio analyzer, and an extant ammonia method. Analytical recovery (y) of drug added to serum (x) was good, with y = 0.97x-0.7, Sy.x = 3.6, r = 0.997 (n = 65) over the range 6.25 to 150 mg/L. Comparison with an HPLC method (x) showed good agreement: y = 0.98x + 1.34, Sy.x = 3.7 (n = 37), as analyzed with the Deming debiased regression. Precision was good, CVs being less than 3% for within-run and less than 6% for between-run controls. Ammonia, amphotericin B, glucose, urea, and hemolysis do not interfere, but bilirubin shows analyte-dependent interference and lipemia interferes when triglycerides exceed 5 g/L. This assay is accurate, inexpensive, and easy to perform. It can be easily adapted for routine or emergency use.

Aminohydrolases↗

Development and evaluation of an automated dye-binding assay for protein in cerebrospinal fluid.

Addition of sodium dodecyl sulfate (SDS) to Coomassie Brilliant Blue reagent equalizes the binding variability of the dye to various proteins and markedly improves the accuracy of quantification of protein in cerebrospinal fluid. In the presence of SDS, the absorption spectrum and the absorption maximum are affected by reaction time and temperature, age of the dye preparation, and protein constituent. I automated this procedure, to optimize precision, enable use of a smaller sample, decrease hands-on time, maintain consistency in the time of reading, and avoid carryover. The results (y) compared well with those of the aca (x), with a Deming de-biased regression equation of y = 0.991x + 14.1 mg/L, Sy.x = 33.4 mg/L. The within-run and between-run precision (CV) was less than 2.5% and less than 4.5%, respectively. Commonly used antibiotics, flucytosine, or amphotericin B do not interfere. This automated procedure is fast and accurate and requires only 10 microL of sample.

Autoanalysis↗

Digital dermal patterns in breast cancer.

Fingerprints of 570 breast cancer cases and the same number of matched controls were obtained from the population-based finger print file in Hawaii for studying the association between breast cancer and digital dermal patterns and ridge counts. The results showed that breast cancer patients had a significant excess of radial loops on the left hand. It was also found that the frequency of ulnar loops on the left hand was significantly elevated for premenopausal women with breast cancer, whereas an excess of radial loops on the left hand was observed for the postmenopausal women with breast cancer. No significant difference for the total and absolute ridge counts was found between breast cancer patients and controls.

Asian People↗

Frequency sensitivities of auditory neurons in the cerebellum of the cat.

Threshold tuning curves were obtained from neurons in the cerebellar auditory area of the cat. The threshold of the brainstem auditory evoked response was also measured in each animal as a function of sound frequency in order to monitor the overall frequency sensitivity of the auditory periphery. Cerebellar auditory neurons responded to sound stimuli with little discrimination for the sound frequency. The values of Q10dB (a measure of the sharpness of tuning) were less than 2 for most of the neurons in this study. There was no significant difference in the sharpness of tuning for neurons in the various layers of the cerebellar auditory area. Electrophysiological mapping showed that the frequency sensitivity of single neurons did not appear to vary as a function of location within the cerebellar auditory area which includes lobules VI and VII of Larsell. Broad tuning was observed in long-latency (greater than 11 ms) neurons which responded to binaural sound stimuli as well as in short-latency (less than 6 ms) neurons which only responded to monaural sound stimuli. Within each animal, tuning curves of single cerebellar neurons were essentially superimposable onto each other and matched well with the overall frequency sensitivity of the animal as shown by brainstem auditory evoked response. Since the frequency tuning of these neurons appeared to reflect the overall frequency sensitivity of the auditory periphery, auditory neurons in the posterior vermis may receive inputs that involve convergence and integration from the entire length of the cochlea.

Animals↗

Tonotopic organization in the inferior colliculus of the rat demonstrated with the 2-deoxyglucose method.

We studied the tonotopic organization in the inferior colliculus of the rat with the 2-deoxyglucose method. Isofrequency bands were observed in the central nucleus of the inferior colliculus. In coronal sections, higher sound frequencies elicited bands that were located more ventrally. At caudal levels of the inferior colliculus, isofrequency bands were relatively short and tilted slightly downward toward the midsagittal plane. As the plane of section moved more rostrally, isofrequency bands gradually lengthened, their orientation first turned horizontally and continued with a downward tilt toward the lateral aspect of the brainstem. At rostral levels, high-frequency (greater than 8000 Hz) bands showed more significant increases in length than low-frequency (less than 4000 Hz) bands. Thus, the amount of tissue in the inferior colliculus devoted to higher frequencies was significantly more than that for lower frequencies. In sagittal sections, the isofrequency band moved from a dorsoposterior position to a ventroanterior one as the plane of section moved more laterally. A three-dimensional model of isofrequency planes was reconstructed from the above data.

Acoustic Stimulation↗

Purification, biochemical characterization, and biological function of human esterase D.

Human esterase D (carboxylesterase; carboxylic-ester hydrolase, EC 3.1.1.1), a genetic marker of retinoblastoma, was purified to biochemical homogeneity from erythrocytes. The purification scheme including carboxymethylcellulose, phenyl-Sepharose, chromatofocusing, and hydroxylapatite chromatographies resulted in a 10,000-fold purification of the enzyme with 15% recovery of total activity. The Km of esterase D was estimated to be 10 X 10(-6) M using 4-methylumbelliferyl acetate as substrate. The enzymatic activity was inhibited by p-chloromercuribenzoate and HgCl2, suggesting an important role of SH group(s) in enzyme function. Specific rabbit polyclonal and mouse monoclonal antibodies against esterase D were prepared and recognized either denatured or native human esterase D protein. Moreover, the polyclonal antibodies immunoprecipitated a polypeptide with a molecular mass of about 33-34 kDa from various cell lines of different mammalian species, indicating that the esterase D protein is highly conserved. The highest levels of this enzyme were found in liver and kidney. Furthermore, the expression of esterase D was enhanced 3-fold in a promonocytic cell line treated with phenobarbital but not with phorbol myristate acetate, suggesting that esterase D may have a role in detoxification. The availability of the homogeneous protein and its specific antibodies allows for cloning of the esterase D gene and facilitates studies of retinoblastomas.

Antibodies↗

Electrophysiological mapping of the auditory areas in the cerebellum of the cat.

We mapped the topographic distribution of auditory evoked potentials over the posterior cerebellum of the cat. Within the posterior vermis, positive-going evoked potentials were primarily recorded in lobule VI and negative ones in lobule VII. Peak latencies of these evoked potentials were between 8 and 18 ms. The amplitudes of evoked potentials from adjacent surface cortical sites often differed significantly. Thus, besides the segregation of positive and negative polarities in the evoked potentials from lobules VI and VII, the distribution of cerebellar auditory evoked potentials within a given lobule contained high- and low-amplitude sites arranged in a patchy pattern. Within the posterior vermis, the evoked potentials showed significant polarity and amplitude changes as a function of depth during microelectrode penetrations which were vertical to the surface of the cerebellar cortex. Cerebellar auditory evoked potentials were not detected over the paramedian lobules and most of the cerebellar hemispheres. There were no significant changes in the electrical activities as a function of depth in these areas. Auditory responses, however, were observed in the most lateral part of the cerebellar hemispheres.

Animals↗

Experimental infection of subperiodic Brugia malayi in laboratory rats with emphasis on evaluation by four techniques.

Infective larvae of subperiodic B. malayi from South Kalimantan (Borneo), Indonesia collected from laboratory-raised Ae. togoi mosquitoes after feeding on infected mongolian gerbils (Meriones unguiculatus) were inoculated subcutaneously into the groin areas of 15 SD and 36 LE rats. Blood was examined weekly by membrane filtration and thick smears starting 10 weeks post-infection. Microfilariae were found in 3 SD and 4 LE rats, the mf infection rate of 20% and 11% respectively. The prepatent period was significantly shorter in the SD rats (99-112 days) than those in the LE rats (110-153 days). The patent period was longer in the LE rats (208-703 days) than in the SD rats (236-543 days), and the mf density was similar (17.5 mf/20 c.mm blood against 16 mf/20 c.mm blood). At necropsy, 6 (3 female and 3 male) adult worms were recovered from 3 of 6 SD rats and 12 (9 female and 3 male) adult worms from 4 of 20 LE rats; all worms were found in the testes. The results of xenodiagnostic, histochemical staining and measuring spicules and protuberances, demonstrated clearly the difference between both species of Brugia. All dissected Ar. subalbatus mosquitoes exposed to B. pahangi became infected (100%), but none of those to subperiodic B. malayi were infected (0%). The mf of both species of Brugia in thick films stained with naphthol-AS-TR-phosphate showed that the excretory and anal pores of subperiodic B. malayi mf exhibited acid phosphatase activity and only a little activity was seen in other parts; while B. pahangi mf showed heavy diffuse acid phosphatase activity along the entire length of the body.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Detection of immunoglobulin heavy chain IgG3 polymorphism in wild mice with xenogeneic monoclonal antibodies.

We have generated four xenogeneic rat antimouse IgG3 monoclonal antibodies recognizing at least three different antigenic determinants (epitopes) on BALB/c IgG3 molecules. These antibodies were used in solid-phase blocking radioimmunoassays for detection of the epitopes in sera of 40 inbred strains and 134 wild mice. These antibodies detect genetic polymorphism of IgG3 isotype among wild mice even though there is no polymorphism found among 40 inbred strains tested (except X-chromosome-linked immunodeficient CBA/N strain which lacks IgG3 molecules). An IgG3 variant was also isolated from hybridomas derived from Mus spretus.

Animals↗

Genetic characterization of mouse immunoglobulin allotypic determinants (allotopes) defined by monoclonal antibodies.

We have generated a new series of monoclonal antibodies recognizing allotypic determinants on mouse IgG1, IgG2a, and IgG2b. In this communication we describe their reactivities with immunoglobulins of the inbred mouse strains. Comparison with serology charts indicates that many of these monoclonal antibodies detect allotypic specificities previously defined by conventional antisera; others define previously undescribed specificities. Strain and isotype distribution allows us to assign five new allotypic specificities to Igh-1 and three new specificities to Igh-3. In addition, on the basis of reactivity with the monoclonal antibodies, we have defined a new Igh haplotype in SWR/J mice, Ighp.

Alleles↗

Structural characterization of mouse immunoglobulin allotypic determinants (allotopes) defined by monoclonal antibodies.

We have generated a new series of monoclonal antibodies recognizing allotypic determinants on mouse IgG1, IgG2a, and IgG2b. In this communication we describe their reactivity at the molecular level. A number of genetic specificities (as defined by reactivity with sera from inbred strains) were divided into subspecificities (allotopes) by these analyses. With the exception of one allotope located in the hinge region of Igh-1b, all other 23 allotopes examined were preserved upon reduction and alkylation of immunoglobulin antigens. To further analyze the role of immunoglobulin conformation in presenting the allotopes, we assayed their presence on mixed Igh-1a/Igh-4a heavy chain molecules. The Igh-1a determinants were maintained, but the Igh-4a determinants were lost. Taken together, our results indicate that genetic polymorphisms at the Igh loci generate an enormous antigenic complexity, much of which relies on tertiary and quaternary protein structure for expression.

Alleles↗

Flow of mitochondrial DNA across a species boundary.

Restriction analysis shows that wild Scandinavian mice belonging to the species Mus musculus contain the mitochondrial DNA of a neighboring species, M. domesticus. This demonstration results from comparisons of Scandinavian mice with authentic M. domesticus and M. musculus from other parts of Europe. Electrophoretic and immunological analysis of eight diagnostic proteins confirms that mice from north of the hybrid zone in Denmark are M. musculus in regard to their nuclear genes. In contrast, the mice tested from this region and a nearby part of Sweden have exclusively M. domesticus types of mitochondrial DNA. Phylogenetic analysis of the restriction maps suggests that the mitochondrial DNAs found in Scandinavian M. musculus could stem from a single M. domesticus female.

Alleles↗

Projections from the cochlear nucleus to the cerebellum.

The connections between the brainstem auditory system and the cerebellar auditory area in the posterior vermis (VI, VII, and VIIIA) were studied using the horseradish peroxidase technique. Injections also were made in the cerebellar hemispheres, the paramedian and the vestibulocerebellum (VIIIB, IX). Labeled cells were found in the cochlear nuclei bilaterally, but not in the superior olivary complex or the inferior colliculus. As the injection sites moved more laterally from the vermis area to the paramedian and the cerebellar hemispheres, the number of labeled cochlear nucleus neurons decreased progressively. This observation suggests that the terminals of the cochlear-nucleus-cerebellum projection tend to concentrate in the vermis auditory area. In the cochlear nucleus, labeled cells did not appear to belong to a single, homogenous cell type, nor did the cells group into a single area in the cochlear nucleus. The total number of labeled neurons in one cochlear nucleus did not exceed 200 in our largest injection.

Animals↗

Trigeminal evoked potentials in the cat.

Electrical stimulation of the infraorbital nerve in the cat resulted in a series of far-field evoked potentials at the vertex. The wave form of these potentials is similar to the auditory brain stem evoked response and the somatosensory far-field response; it is multicomponent, the amplitudes of individual components are of the order of 1 microV, and the latencies are all less than 4 msec. The anatomical origins of these evoked potentials were investigated. The results indicated that contributions due to the afferent trigeminal nerve, the principal sensory nucleus, and the spinal trigeminal nucleus are involved.

Animals↗