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Biomedical subjects

H N Chang

Publications and source records attributed to H N Chang.

At least 37 records · Page 2Linked to original sources
PubMed1996

Angiogenic induction and cell migration in an orthopaedically expanded maxillary suture in the rat.

The purpose was to examine the effect of an angiogenic factor on cell migration patterns and osteoblast histogenesis during the 96 h following orthopaedic expansion of the anterior maxillary suture. Fifty rats were divided into four groups: (1) a control group that received only angiogenic induction via injection of 5 ng/g body wt recombinant human endothelial-cell growth factor; (2) an experimental group that received orthopaedic expansion and angiogenic induction; (3) a sham group that received orthopaedic expansion and normal saline injection; and (4) a baseline group that received no expansion or injection. The experimental and sham groups were subdivided to conduct experiments over 1, 2, 3 or 4 days. The anterior portion of each maxilla was dissected free and demineralized. Sections (4 microns thick) were cut from every block and stained with Mayer's haematoxylin and eosin. Cell migration was analysed using a previously established cell-kinetics model. The osteoprogenitor cells were divided into four categories according to nuclear volume: A cells (40-79 microns3), B cells (80-119 microns3), C cells (120-169 microns3) and D cells (> or = 169 microns3 A' cells are the portion of the A cell population that responds to osteogenic stimulus. As previously defined in periodontal ligament, the reciprocal association of a decreasing number of less differentiated (A + A) cells and an increasing number of C + D cells, as a function of distance from the nearest major blood vessel, was consistently found in all groups. This suggests a vascularly oriented gradient of progressively more differentiated osteoprogenitor cells. Also, A + A' cells were predominately located within 20 microns of the nearest major blood vessel whereas the C + D cells were found at a distance > 30 microns from the nearest major blood vessel. These results suggest that the A'-->C shift occurs 20-30 microns from the nearest major blood vessel. In the angiogenic induction groups, the numbers of committed osteoprogenitors (A + A') were significantly higher than in the sham group at day 1. At day 3, the numbers of preosteoblasts (C + D) in angiogenic sutures were significantly higher than in the sham groups. This enhancement of preosteoblast population strongly suggests the possible role of activated pericytes in expanded sutures as a source of osteoprogenitor cells.

Analysis of Variance↗
PubMed1996

The relationship of aging to endotoxin shock and to production of TNF-alpha.

BACKGROUND: Aged people are considered prone to gram-negative bacteremia and septic shock. This relationship was tested in murine endotoxin shock. METHODS: Balb/c mice of various ages (1.4-13.4 months) were intraperitoneally injected with lipopolysaccharide (LPS), and rates of survival were observed. The production of TNF-alpha in vivo induced by LPS was measured. RESULTS: The survival rates were the smallest in the oldest and youngest groups. Production of TNF-alpha attained a maximum at 2 h after LPS injection and was smaller in the oldest group; it had a reciprocal relationship to survival rates in each group except the youngest group. CONCLUSION: Old and young mice had smaller rates of survival and greater production of TNF-alpha following endotoxin shock induced by LPS.

Aging↗
PubMed1995

Production of poly(hydroxyalkanoic acid).

Poly(hydroxyalkanoic acid) [PHA] is accumulated by numerous microorganisms as an energy reserve material under unbalanced growth conditions in the presence of excess carbon source. In spite of being a good candidate for biodegradable thermoplastics, their high price compared with conventional plastics currently in use has limited their availability in a wide range of applications. With the aim of reducing the high production cost of PHA, much effort is currently being devoted to improve productivity by employing various microorganisms and by developing efficient culture techniques. Several processes recently developed and employed for the production of PHA by various bacteria are described.

Acyltransferases↗
PubMed1995

Production of poly(3-hydroxybutyric acid) by recombinant Escherichia coli strains: genetic and fermentation studies.

A number of Escherichia coli strains including K12, B, W, XL1-Blue, DH5 alpha, HB101, JM109, and C600 were transformed with the stable high-copy-number plasmid pSYL105 containing the Alcaligenes eutrophus polyhydroxyalkanoic acid biosynthesis genes, and were subsequently compared for their ability to synthesize and accumulate poly(3-hydroxybutyric acid) (PHB). The rate of PHB synthesis, the extent of PHB accumulation, and PHB yield from glucose varied considerably from one strain to another. Strains XL1-Blue and B harboring pSYL105 synthesized PHB at the highest rate to a final concentration of ca. 7 g/L in complex medium containing 20 g glucose/L. With an aim to reduce the cost of the medium, the effect on PHB accumulation of supplementing a defined medium with complex nitrogen sources was examined. A PHB concentration of 81 g/L could be obtained in 41 h from a pH-stat fed-batch culture of XL1-Blue(pSYL105) in a semidefined medium. When the availability of acetyl-CoA was increased by supplementing the medium with complex nitrogen sources, amino acids, or oleic acid, PHB synthesis by recombinant E. coli was enhanced.

Alcaligenes↗
PubMed1995

Hypersensitivity to mosquito bite: a case report.

Severe hypersensitivity to mosquito bites is very rare. A 18-year-old girl student has had multiple hemorrhagic bullae with atrophic scars on both limbs and left side of the face due to mosquito bites for 16 years. When she was seven years old, she noticed a severe local reaction with bloody bullae formation, chills, and fever several hours after the mosquito bite. The bullae developed into ulcers which finally became scars. This patient was confirmed to be hypersensitive to mosquito bites by the mosquito bite test. This case is the first record of mosquito allergy in Taiwan.

Adolescent↗
PubMed1994

Shikonin production by extractive cultivation in transformed-suspension and hairy root cultures of Lithospermum erythrorhizon.

Effects of in situ extraction, fungal elicitation, a permeabilizing agent, and the oxygen transfer rate on shikonin production in transformed suspension and hairy root cultures of Lithospermum erythrorhizon were studied. Shikonin production with in situ extraction in transformed cell and hairy root cultures by n-hexadecane was 7.6 and 3 times higher than those of the control culture. Shikonin production of transformed L. erythrohizon increased with the enhanced gas exchange, and in situ extraction also increased sucrose consumption and shikonin production. The optimal volume of n-hexadecane in the hairy root culture was similar to that in the transformed cell cultures. In situ extraction at an earlier stage significantly enhanced shikonin production both in transformed cell and hairy root cultures. Dimethylsulfoxide used as a permeabilizing agent was harmful to cell growth and shikonin production, and permeabilizing was unnecessary when in situ extraction was applied. This occurred because with the solvent addition, most shikonin was spontaneously released from the cells and was dissolved in the solvent layer. The combined addition of n-hexadecane of the extract of the fungus Penicillium as an elicitor seemed to result in a higher production of shikonin both in cell suspensions and transformed root cultures. However, an increase of shikonin induction by fungal elicitation in a hairy root culture was not significant in comparison with that of normal cell cultures.

Agrobacterium tumefaciens↗
PubMed1994

Construction of plasmids, estimation of plasmid stability, and use of stable plasmids for the production of poly(3-hydroxybutyric acid) by recombinant Escherichia coli.

Plasmids containing the Alcaligenes eutrophus poly(3-hydroxybutyric acid) (PHB) biosynthetic genes were constructed for the production of PHB in Escherichia coli and plasmid stability was investigated by repeated subculturing without antibiotic pressure. Both pSYL101 (high copy) and pSYL102 (medium copy) were unstable during the subcultures. Higher instability was observed when cells were accumulating PHB. Segregational instability was aggravated by the faster growth of plasmid-free cells and by appearance of non-dividing cells harboring large amount of PHB during the fed-batch culture. Two derivatives, pSYL103 and pSYL104, were then developed by cloning the parB locus of plasmid R1 into pSYL102 and pSYL101, respectively. They showed 100% stability even during PHB synthesis and accumulation over 110 generations. All four plasmids were structurally stable. The final cell mass, PHB concentration, and PHB per dry cell weight (P/X, w/w, %) of 101.4 g l-1, 81.2 g l-1, and 80.1%, respectively, were obtained in 39 h by high cell density culture of XL1-Blue (pSYL104). The final PHB concentration was lower using XL1-Blue (pSYL103), which suggested that high gene dosage was required for the synthesis and accumulation of PHB to a high concentration in E. coli.

Escherichia coli↗
PubMed1994

High density cell culture by membrane-based cell recycle.

Enhancement of productivity of a bioprocess necessitates continuous operation of bioreactors with high biomass concentrations than are possible in conventional batch, fedbatch or continuous modes of culture. Membrane-based cell recycle has been effectively used to maintain high cell concentrations in bioreactors. This review compares membranebased cell recycle operation with other such high density cell culture systems as immobilized cell reactors and reactors with cell recycle by centrifugation or gravity sedimentation. A theoretical of production of primary and secondary metabolites in membrane-based recycle systems is presented. Operation of this type of system is discussed with examples from aerobic and anaerobic fermentations.

Journal Article↗
PubMed1994

Immunoblot analysis of components of Penicillium citrinum recognized by human IgE and IgG antibodies.

Penicillium species are well-known indoor fungi and have been considered as one of the causative agents of extrinsic bronchial asthma. We have identified that Penicillium citrinum is the most prevalent Penicillium species in the Taipei area. In the present study, components of this airborne fungus recognized by human IgE and IgG antibodies were analyzed by immunoblotting. Eight components ranging in MWs from 112 to 17.5 kD reacted with IgE antibodies in 11 serum samples. The component with an MW of about 33 kD showed the highest frequency (81.8%) of IgE-binding. However, 11 components of P. citrinum showed IgG-immunoblotting activity, and the component with an MW of about 66 kD revealed the highest frequency (58.3%) of IgG-binding. Only 3 of the 9 serum samples which reflected IgE antibody activity to the 33 kD component of P. citrinum showed IgG-immunoblotting activity to the same 33 kD component. Furthermore, 4 of the 5 serum samples obtained from healthy laboratory personnel demonstrated IgG responses, but none had IgE antibody activity to components of p. citrinum. The discrepancy observed between IgE and IgG responses to the same allergenic extract requires further investigation.

Binding Sites↗
PubMed1993

Production of monoclonal antibodies against human chorionic gonadotropin by hybridoma cultures in calcium alginate capsules.

A new method of making microcapsules with calcium alginate gel was developed and the cultivation of the encapsulated hybridoma cells producing monoclonal antibodies against human chorionic gonadotropin was investigated. A high cell density of 2.0 x 10(8) cells/cm3 in the capsules led to a high dilution rate of 0.67 per hour and resulted in the high volumetric monoclonal antibody productivity of 652.8 mg/l/day, which was 20-30 times higher than those of traditional continuous suspension cultures. However, long-term continuous culture was not achieved with this capsule system probably because of the limitation in nutrient supply and the accumulation of waste products. Also the analysis of oxygen transfer in this system showed that oxygen supply was not enough to support such a high cell density.

Alginates↗
PubMed1993

Bench-scale production of acrylamide using the resting cells of Brevibacterium sp. CH2 in a fed-batch reactor.

Effects of various organic acids and salts on the stabilization of nitrile hydratase were investigated. The stability of the nitrile hydratase of Brevibacterium CH2 during storage was greatly enhanced by the addition of n-butyric acid. Effects of temperature, pH, and concentrations of acrylonitrile and n-butyric acid on acrylamide production by the resting cells were also investigated. Acrylamide production per unit dry weight of the cells increased 1.33 times by the addition of 0.05% n-butyric acid. A 20% acrylamide solution was successfully produced in a bench-scale reactor (12 l) with only a trace amount of salts after 10 h of hydration reaction under optimum reaction conditions without using an isotonic substrate. The conversion yield was nearly 100%, and acrylic acid as a by-product was not produced. Final acrylamide production of 400 g g-1 cells and productivity of 20 g/(g cells l-1 x h-1) were obtained.

Acrylamide↗
PubMed1992

Continuous production of 6-APA in an aqueous two-phase system.

An aqueous two-phase system of polyethylene glycol (PEG) and potassium phosphate provided a favorable environment for bioconversion of penicillin G to 6-aminopenicillanic acid (6-APA). The recombinant E. coli cells containing penicillin acylase were partitioned in the phosphate-rich bottom phase, and the product 6-APA in the PEG-rich top phase, which protected the enzyme from deactivation for a longer period of time. The continuous production of 6-APA by recycling the bottom phase showed a slight decrease in enzyme activity from initial 80 mM of 6-APA production to 61 mM during the eight days of operation at a space velocity of 0.12 hours-1. This work showed a possibility that the aqueous two-phase whole-cell enzyme bioconversion could serve as an alternative to immobilized cells on solid matrices.

Enzymes, Immobilized↗
PubMed1991

Membrane bioreactors: present and prospects.

Membrane bioreactors have a very handy in-situ separation capability lacking in other types of bioreactors. Combining various functions of membrane separations and biocatalyst characteristics of enzymes, microbial cells, organelles, animal and plant tissues can generate quite a number of membrane bioreactor systems. The cell retaining property of membranes and selective removal of inhibitory byproducts makes high cell density culture possible and utilizes enzyme catalytic activity better, which leads to high productivity of bioreactors. Enzyme reactions utilizing cofactors and hydrolysis of macromolecules are advantageous in membrane bioreactors. Anaerobic cell culture may be efficiently carried out in membrane cell recycle systems, while aerobic cultures work well in dual hollow fiber reactors. Animal and plant cells have much a better chance of success in membrane reactors because of the protective environment of the reactor and the small oxygen uptake rate of these cells. Industrial use of these reactors are still in its infancy and limited to enzyme and animal tissue culture, but applications will expand as existing problems are resolved.

Animals↗
PubMed1991

Effects of periodic backflushing on ultrafiltration performance.

Periodic backflushing was introduced to a membrane separation process to improve the performance. Hemoglobin (M.W. = 62,500) and dextran (M.W. = 10,000) were used as model compounds. Filtration performance of an ultrafiltration membrane system (Amicon hollow fiber membrane, H1P30-43, molecular weight cutoff = 30,000) was measured in terms of apparent permeability and retention coefficient of dextran to determine the effects of backflushing frequency and duration of one cycle. An optimum frequency around 0.2 min-1 existed to give a maximum permeability while the retention of dextran decreased with increasing frequencies. The improvement in permeability by periodic backflush was more than doubled. The retention of dextran decreased as backflushing duration was increased in one cycle. With the duration of 33.75 s, the retention of dextran was less than 50% and dextran output was 1.14 g/h, which was 1.3 times the value without backflushing. Also, periodic backflush made possible the long-term filtration of yeast cells for more than 20 h.

Dextrans↗