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Biomedical subjects

K Gao

Publications and source records attributed to K Gao.

35 records · Page 2Linked to original sources
PubMed1999

Nucleic acid duplexes incorporating a dissociable covalent base pair.

We have used molecular modeling techniques to design a dissociable covalently bonded base pair that can replace a Watson-Crick base pair in a nucleic acid with minimal distortion of the structure of the double helix. We introduced this base pair into a potential precursor of a nucleic acid double helix by chemical synthesis and have demonstrated efficient nonenzymatic template-directed ligation of the free hydroxyl groups of the base pair with appropriate short oligonucleotides. The nonenzymatic ligation reactions, which are characteristic of base paired nucleic acid structures, are abolished when the covalent base pair is reduced and becomes noncoplanar. This suggests that the covalent base pair linking the two strands in the duplex is compatible with a minimally distorted nucleic acid double-helical structure.

Base Pairing↗
PubMed1999

An in vitro study of wheat bran binding capacity for Hg, Cd, and Pb.

Water-soluble dietary fiber (WSDF), water-insoluble dietary fiber (WIDF) from wheat bran, and the carboxymethylated product of WIDF (CIDF), all having low contents of protein, ash, and phytic acid, were evaluated for their scavenging capacity for three heavy metals, Hg, Cd, and Pb. The results showed that WIDF had higher BC(max) (maximum amount of bound heavy metal ions) and BC(min) values (minimum concentration of heavy metal ions below which the ions cannot be bound by dietary fibers) than WSDF at two pH conditions (pH 2.0 and 7.0). Carboxymethylation of WIDF improved its binding capacity for heavy metals (increase in BC(max) and decrease in BC(min)). The pH value significantly affected the binding capacity for heavy metals; BC(max) sharply increased and BC(min) sharply decreased for each heavy metal ion for all of the dietary fibers when the pH was raised from 2.0 to 7.0. The binding capacity of dietary fibers for heavy metals was slightly affected by amino acids, calcium, iron, and zinc but significantly affected by copper. Colon fermentation released part of the heavy metal ions from dietary fibers. From the results it can be concluded that dietary fibers from wheat bran can effectively bind all three tested metal ions to prevent the body from being affected by their toxicity.

Cadmium↗
PubMed1999

Lignan and phenylpropanoid glycosides from Lancea tibetica and their antitumor activity.

A new lignan glucoside, the first 7,9'-monoepoxyte-trahydrofuran type lignan with the cis-relationship of H-7 and H-8, named tibeticoside (1), as well as ten known compounds have been isolated from the medicinal plant (roots, stems and leaves) Lancea tibetica. The structure of tibeticoside (1) has been elucidated on the basis of chemical and spectral evidence, especially by 2D-NMR (1H-1H COSY, HMQC, HMBC, NOESY). In addition, sylvatesmin (6) exhibited effective antitumor activity on B16 cells.

Animals↗
PubMed1998

Activation of serotonergic neurons in the raphe magnus is not necessary for morphine analgesia.

A wealth of pharmacological and behavioral data suggests that spinally projecting serotonergic cells mediate opioid analgesia. A population of medullary neurons, located within raphe magnus (RM) and the neighboring reticular nuclei, contains serotonin and is the source of serotonin in the spinal dorsal horn. To test whether serotonergic neurons mediate opioid analgesia, morphine was administered during recordings from medullary cells that were physiologically characterized as serotonergic (5HTp) by their slow and steady discharge pattern in the lightly anesthetized rat. Selected 5HTp cells (n = 14) were intracellularly labeled, and all contained serotonin immunoreactivity. The discharge of most 5HTp cells was not affected by an analgesic dose of systemic morphine. In a minority of cases, 5HTp cells either increased or decreased their discharge after morphine administration. However, morphine altered the discharge of some 5HTp cells in the absence of producing analgesia and conversely did not alter the discharge of most 5HTp cells in cases in which analgesia occurred. RM cells with irregular discharge patterns and excitatory or inhibitory responses to noxious tail heat were classified as ON and OFF cells, respectively. All ON and OFF cells that were intracellularly labeled (n = 9) lacked serotonin immunoreactivity. All ON cells were inhibited, and most OFF cells were excited by systemic morphine. Because 5HTp cells do not consistently change their discharge during morphine analgesia, they are unlikely to mediate the analgesic effects of morphine. Instead, nonserotonergic cells are likely to mediate morphine analgesia in the anesthetized rat. In light of the sensitivity of morphine analgesia to manipulations of serotonin, serotonin release, although neither necessary nor sufficient for opioid analgesia, is proposed to facilitate the analgesic effects of nonserotonergic RM terminals in the spinal cord.

Action Potentials↗
PubMed1997

Somatodendritic and axonal anatomy of intracellularly labeled serotonergic neurons in the rat medulla.

A knowledge of the anatomy of medullary serotonergic cells is critical to understanding local and brainstem circuits in which these cells participate. Serotonergic neurons (n = 16) were identified, as previously described (Mason [1997] J. Neurophysiol. 77:1087-1098) by their slow and steady background discharge in halothane anesthetized rats. Neurons were then intracellularly labeled with Neurobiotin and visualized with 3,3'diaminobenzidine. The validity of the physiological identification of serotonergic cells was confirmed by processing two neurons that were physiologically characterized as serotonergic for serotonin immunoreactivity; both tested cells contained immunoreactive serotonin. The dendrites and axon of each labeled cell were reconstructed by using a three-dimensional computerized system. Somata were small or medium in size and had fusiform, triangular, or multipolar shapes. The dendritic arbor was constricted with most dendrites extending for less than 500 microm from the soma. All labeled axons projected caudally and travelled in the ventrolateral medulla, either dorsal or ventral to the lateral reticular nucleus. Most cells had collaterals and/or dense axonal swellings in the nucleus reticularis gigantocellularis, nucleus reticularis magnocellularis, raphe magnus, and the ventrolateral medulla. Non-local collaterals and swellings were also observed in the nucleus reticularis gigantocellularis and in the ventrolateral medulla at all medullary levels. The results demonstrate that 1) the dendrites of serotonergic cells are restricted to raphe magnus and the ventral part of nucleus reticularis magnocellularis; and 2) serotonergic cells project to medullary nuclei that contain bulbospinal cells which project to dorsal, intermediate, and ventral horns. Serotonergic cell projections to brainstem sites may mediate the integration of sensory, autonomic, and motor modulation at the brainstem level.

Animals↗
PubMed1997

SEROTONERGIC pontomedullary neurons are not activated by antinociceptive stimulation in the periaqueductal gray.

The antinociceptive and cardiovascular effects of midbrain periaqueductal gray (PAG) stimulation are mediated through a relay in the pontomedullary raphe magnus (RM) and adjacent nucleus reticularis magnocellularis (NRMC). To test whether the neurons important in mediating PAG-evoked effects are SEROTONERGIC, the responses of pontomedullary SEROTONERGIC-LIKE cells to PAG stimulation were tested. SEROTONERGIC-LIKE neurons (n = 21) were recorded extracellularly in halothane-anesthetized Sprague Dawley rats. Serotonergic-like neurons were distinguished by their slow and steady background discharge. Two neurons that were physiologically characterized as SEROTONERGIC-LIKE were intracellularly labeled and processed for serotonin immunoreactivity; both cells tested contained immunoreactive serotonin. Train stimulation of sites within the midbrain PAG, at intensities of </=50 microA, suppressed the tail withdrawal from noxious heat and evoked changes in blood pressure and heart rate. No SEROTONERGIC-LIKE cells were activated by single-pulse or short-train (two to five pulses) stimulation of the PAG at antinociceptive intensities. In most cases, SEROTONERGIC-LIKE cells were unaffected by long-train stimulation (5-6 sec) of the PAG, which produced antinociception and cardiovascular changes. In contrast, >50% of the cells in two nonserotonergic-like cell classes were activated at short latency by such PAG stimulation. In conclusion, monosynaptic excitation of SEROTONERGIC cells in RM/NRMC is unlikely to be necessary for the nociceptive and autonomic modulatory effects of PAG stimulation.

Animals↗
PubMed1997

Study on the hydrolyzate of sugarcane bagasse to ethanol by fermentation.

Study of glucose and xylose utilization by Pichia stipitis in a limited oxygen supply condition revealed that the rate of glucose utilization decreased rapidly while that of xylose decreased slowly until the time that glucose and xylose were shown to level out, at which point the rate of xylose utilization increased rapidly. Based on the results, ethanol fermentation technology in continuous connective tower fermenters was advanced, e.g., fermentation by P. stipitis in an airlift loop tower focusing on xylose utilization and then residue glucose utilization by Saccharomyces cerevisiae in an overflow tower. When the fed hydrolyzate of bagasse was concentrated in five folds and the dilution rate was 0.1 h-1, the total utilization ratio of reducing sugar was 97.2%; the concentration of ethanol was 46.4 g/L.h.

Cellulose↗
PubMed1996

[Surgical treatment and classification of spontaneous pneumothorax caused by break-up of bullae].

OBJECTIVE: To explore the appropriate operation for surgical treatment and a better classification for spontaneous pneumothorax caused by break-up of bullae. METHODS: Eighty two cases of spontaneous pneumothorax caused by break-up of bullae were reported and classified by their clinical characteristics and manifestations during operation. RESULTS: All cases were classified into type I (apical), type II (lobular), type III (diffusive) and type IV (bilateral) on the basis of bullae's appearance and location. And the cases of type I were further classified into Ia (simple) and Ib (pneumohemothoracic). CONCLUSION: The clinical classification was thought to be valuable for diagnosis and treatment of spontaneous pneumothorax caused by break-up of bullae.

Adolescent↗
PubMed1995

Epipolarization microscopic immunogold assay: a combination of immunogold silver staining, enzyme-linked-immunosorbent assay and epipolarization microscopy.

We describe a new immunoassay which combines an immunosorbent assay, immunogold silver staining and epipolarization microscopy. Our new assay procedure features multiple samples on a single microscope slide, and high sensitivity of epipolarization microscope for detection of silver-enhanced colloidal gold as a final immunoassay product. We call the new immunoassay "on slide immunogold assay" (OSIGA). This new method uses biotinylated antibody and streptavidin-gold reaction with silver enhancement technique. With OSIGA it is possible to investigate 30 samples on a single microscopic slide. Our preliminary studies used 10-20 microliter samples and detected nanogram quantities of a standardized protein solution. Unlike enzyme linked immunosorbent assay (ELISA), which has a limited time for reading the final color products, the OSIGA specimens can be dried or resin mounted for longer storage and future reference.

Animals↗
PubMed1994

Optimal visualization of immunogold-silver staining of hepatic PEPCK with epipolarized light microscopy.

We used immunogold-silver staining to localize phosphoenolpyruvate carboxykinase in 10 microns cryosections of 4% paraformaldehyde perfusion-fixed normal male rat liver. The resolution and sensitivity of detection were improved by epipolarized light microscopy of 0.5 microns semi-thin plastic sections prepared from these pre-embedding immunogold-silver-enhanced 10-microns thick cryosections. Epipolarized light combined with transmitted light simultaneously demonstrated antigenic sites (visualized with epipolarized light illumination) and tissue morphology (revealed by transmitted light). To optimize the conditions for high resolution, an oil immersion objective lens (x 100) with adjustable iris diaphragm was used with different intensity settings for both light sources. Our observations indicate that if the intensity of the transmitted light is too high, the visibility of the gold-cored silver grains by epipolarized illumination is decreased; if the intensity of epipolarized light is too strong, haloes appear around the gold-cored silver particles. By adjusting the aperture in the objective lens and the neutral density filter in the transmitted light pathway to balance the intensities of transmitted and epipolarized light, an optimal image is obtained that shows the maximal number of antigenic sites and excellent morphology.

Animals↗
PubMed1993

Immunogold-silver staining and epipolarized light microscopic detection of phosphoenolpyruvate carboxykinase and glycogen phosphorylase in rat liver.

The subcellular distribution of enzymes related to carbohydrate metabolism was determined in sections of paraformaldehyde fixed and polyethylene glycol-1540-embedded rat liver and in cryostat sections. For this purpose, goat anti-rat phosphoenolpyruvate carboxykinase (PEPCK) serum and rabbit anti-rat glycogen phosphorylase (GP) serum were used as primary antibodies to localize the corresponding antigens. The primary antibodies were localized by 5 nm colloidal gold labeled secondary antibodies (either rabbit anti-goat IgG for PEPCK or goat anti-rabbit IgG for GP), and the gold particles were enhanced by silver staining using appropriate development reagents. The silver enhanced gold particles were detected by epipolarized light microscopy. PEPCK and GP immunoreactive molecules were found only in glycogen-containing areas of the cytosome of hepatocytes, and not in other cells. No immunocytochemical staining of hepatocytes was found when normal serum replaced the primary antibody in the procedures. Visio-Bond semithin (0.35-1.0 micron) sections provided higher resolution for subcellular immunostaining of PEPCK and GP than cryosections of 10 microns. Epipolarized light microscopy provided detection at high sensitivity of the gold-labeled antibody, and combined with transmitted light, allowed simultaneous visualization of the tissue morphology.

Animals↗
PubMed1993

Enhancement of antigenic site detection with gold labeled secondary and tertiary antibodies using the immunogold-silver staining method.

We report a modification of the immunogold-silver staining method (IGSS) for localizing hepatic phosphoenolpyruvate carboxykinase (PEPCK) in tissue sections, and we compare the efficacy of localizing the primary antibody with either a 5 nm gold labeled secondary antibody or 5 nm gold labeled secondary and tertiary antibodies. Light microscope examination of 10 microns frozen sections demonstrated that the use of combined secondary and tertiary gold labeled antibodies was superior to using a secondary gold labeled antibody alone. The increased labeling density (number of colloidal gold particles/antigenic site/cell) achieved by combined gold labeled antibodies was confirmed by electron microscopy. The increased labeling density resulted in a two-thirds reduction in the time needed for the IGSS physical development of the silver shells and less background. We achieved intense specific staining of hepatocytes expressing PEPCK while minimizing background staining. The use of combined secondary and tertiary gold labeled antibodies enhances the signal-to-noise ratio, achieves high resolution and is a suitable method for use in both light and electron microscopy.

Animals↗
PubMed1987

Solid core liposomes with encapsulated colloidal gold particles.

Solid core liposomes with encapsulated colloidal gold particles were prepared through four major steps: Preparation of prevesicles with encapsulated solid cores of agarose-gelatin by emulsification of agarose-gelatin sol in organic solvent containing emulsifiers followed by cooling. Extraction of lipophilic components from prevesicles to obtain microspherules of agarose-gelatin. Introducing colloidal gold particles into microspherules and coating with protein molecules. Encapsulation of colloidal gold-bearing microspherules with the modified organic solvent spherule evaporation method for preparation of liposomes (Kim et al. (1983) Biochim. Biophys. Acta 728, 339-348 and Kim et al. (1984) Biochim. Biophys. Acta 812, 793-801). Electron micrographs showed that if liposomes were prepared by using a lipid mixture containing dioleoylphosphatidylcholine/cholesterol/dioleoylphosphatidylglycerol/tri olein (molar ratio 4.5:4.5:1:1), there was only a single continuous bilayer membrane for each solid core liposome. However, if no triolein was added to the lipid mixture, it would cause the formation of multilamellar liposomes. In both cases, there were hundreds to thousands of colloidal gold particles within each solid core liposome.

Cholesterol↗