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Biomedical subjects

W J Huang

Publications and source records attributed to W J Huang.

At least 19 recordsLinked to original sources
PubMed1997

Characterization of poly(glycolide-co-D,L-lactide)/poly(D,L-lactide) microspheres for controlled release of GM-CSF.

PURPOSE: This study describes the preparation and characterization of a controlled release formulation of granulocyte-macrophage colony-stimulating factor (GM-CSF) encapsulated in poly(glycolide-co-D,L-lactide) (PLGA) and poly(D,L-lactide) (PLA) microspheres. METHODS: GM-CSF was encapsulated in PLGA/PLA microspheres by a novel silicone oil based phase separation process. Several different blends of PLGA and low molecular weight PLA were used to prepare the microspheres. The microspheres and the encapsulated GM-CSF were extensively characterized both in vitro and in vivo. RESULTS: Steady release of GM-CSF was achieved over a period of about one week without significant "burst" of protein from the microspheres. Analysis of microsphere degradation kinetics by gel permeation chromatography (GPC) indicated that low molecular weight PLA enhanced the degradation of the PLGA and thereby affected release kinetics. GM-CSF released from the microspheres was found to be biologically active and physically intact by bioassay and chromatographic analysis. Analysis of serum from mice receiving huGM-CSF indicated that the GM-CSF was biologically active and that a concentration of greater than 10 ng/mL was maintained for a period lasting at least nine days. MuGM-CSF was not detected following in vivo administration of muGM-CSF microspheres. The tissues of mice receiving muGM-CSF microspheres were characterized by infiltration of neutrophils, and macrophages which were in significant excess of those found in mice administered with placebo controls (i.e. microspheres without GM-CSF). CONCLUSIONS: This study demonstrates the influence of formulation parameters on the encapsulation of GM-CSF in PLGA/PLA microspheres and its controlled release in biologically active form. The intense local tissue reaction in mice to muGM-CSF microspheres demonstrates the importance of the mode of delivery on the pharmacologic activity of GM-CSF.

Animals↗
PubMed1996

Alterations in pulmonary function after retroperitoneoscopic surgery.

OBJECTIVE: To evaluate and compare changes in pulmonary function after retroperitoneoscopic and open surgery. PATIENTS AND METHODS: From June 1994 to October 1995. 11 patients (five men and six women, mean age 44.7 years, range 29-69) underwent retroperitoneoscopic procedures (Group 1) and 11 patients (eight men and three women, mean age 57.5 years, range 22-73) underwent flank-incision procedures (Group 2). The surgery comprised eight adrenalectomies and one each of nephroureterectomy, nephrectomy and pyelolithotomy in Group 1, and five adrenalectomies, four nephroureterectomies, one ureterolithotomy and one excision of a retroperitoneal tumour in Group 2. Pulmonary function tests (PFTs) were performed before and 3 days after surgery; the forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), forced expiratory volume at 25% (FEV25%), FEV1/FVC, vital capacity (VC), total lung capacity (TLC), residual lung volume (RV) and functional residual capacity (FRC) were compared between the groups. The post-operative changes in the PFTs were assessed using a paired t-test and the degree of change in both groups compared using the Mann-Whitney U-test. Other factors possibly influencing PFT were analysed using multiple regression. RESULTS: Pulmonary function was impaired in both groups on the third day after surgery. In Group 2, the FVC, FEV1, FEV25%, VC and TLC had declined significantly (all P < 0.05) from the pre-operative value. In Group 1, only the FVC and VC decreased significantly (P < 0.05). Post-operative pulmonary complications occurred in two patients in Group 2 but in none of those in Group 1, showing that pulmonary function was generally less affected in Group 1 than in Group 2. CONCLUSION: Although pulmonary function was impaired 3 days after surgery in both groups, retroperitoneoscopic surgery, by eliminating a large flank incision, caused less post-operative pulmonary dysfunction than open surgery.

Adult↗
PubMed1996

Laparoscopic ligation of bilateral spermatic varices under epidural anesthesia.

Feasibility and safety of laparoscopic ligation of bilateral internal spermatic varices under epidural anesthesia were assessed in 11 patients. Another 11 patients undergoing the same procedure under ventilation-assisted anesthesia served as controls. Patients in both groups belonged to the American Society of Anesthesia functional class I. Arterial blood analyses were obtained (1) in the horizontal supine position; (2) in the 15 degrees Trendelenburg position; (3) at 15 min after carbon dioxide pneumoperitoneum insufflation, and (4) at 15 min after desufflation in the supine position. In the epidural anesthesia group arterial blood parameters and respiratory rate remained stable in the Trendelenburg position. After intraperitoneal insufflation of carbon dioxide for 15 min, the arterial carbon dioxide level increased from 40.1 +/- 2.2 to 42.1 +/- 2.6 mm Hg, the respiratory rate increased from 17.0 +/- 1.4 to 20.6 +/- 1.2/min, the blood pH value decreased from 7.386 +/- 0.027 to 7.355 +/- 0.034, all values showing statistically significant differences. These changes returned to the preinsufflation level 15 min after release of the pneumoperitoneum. The above-mentioned parameters remained unchanged under the pneumoperitoneum by assisted ventilation in the control group. The mean time of surgery was similar in both groups: 82 and 90 min for the groups having general and epidural anesthesia, respectively. All laparoscopic procedures were accomplished successfully under general anesthesia. However, failure to ligate the internal spermatic varices occurred in 3 patients under epidural anesthesia, mainly due to patient intolerance to abdominal distension. The operation was continued under intubated general anesthesia for relaxing the abdominal muscle to provide an adequate working space. In 8 patients being successfully operated under epidural anesthesia, 5 experienced mild but tolerable abdominal distension; 2 complained of shoulder pain intraoperatively. Although laparoscopic ligation of internal spermatic varices could be accomplished in some patients under epidural anesthesia, it carried a high failure rate, more intraoperative morbidity, and significant arterial blood gas alterations. Routine ventilation-assisted anesthesia is suggested for therapeutic laparoscopy even for an easy procedure such as the ligation of the internal spermatic varices.

Anesthesia, Epidural↗
PubMed1995

Direct needle insufflation for pneumoretroperitoneum: anatomic confirmation and clinical experience.

OBJECTIVES: The feasibility and safety of direct needle insufflation to create pneumoretroperitoneum was assessed by an imaging study and clinical experience. METHODS: A total of 10 patients without previous retroperitoneal surgery or diseases received computed tomography scans of the retroperitoneum 2 cm above the iliac crest. Distances between quadratus lumborum and colon (Q-C distance) were measured in the supine and lateral positions. Changes of Q-C distance were calculated when the patient was changed from the supine to the lateral position. Operative charts on 38 retroperitoneoscopic procedures were collected prospectively to assess complications related to direct needle insufflation, which was performed by inserting a 14 G Veress needle blindly along the posterior axillary line 2 cm above the iliac crest. RESULTS: Q-C distance increased from 8.7 to 27.3 mm (left side) and 4.6 to 18.1 mm (right side) when the patient was changed from the supine to the lateral position, both P values < 0.05. An average distance of 23 mm between colon and quadratus lumborum was found when patients were lying laterally. The misplacement of a Veress needle was encountered in 1 patient, in which a prefascia insufflation resulted in conversion of the endoscopic procedure. Needle puncture caused no visceral or great vessel injury. CONCLUSIONS: Significant anterior movement of the colon was found when patients were changed from the supine to the lateral position. It provided a window for inserting the Veress needle blindly into the retroperitoneum. The high success rate (97%) and low complication rate of direct needle insufflation were found in actual clinical applications. We considered needle insufflation a safe and effective method of establishing a pneumoretroperitoneum for any retroperitoneoscopic procedure.

Adult↗
PubMed1994

The association of aldosterone-producing adrenal adenoma in a patient with autosomal dominant polycystic kidney disease.

We report a young female with autosomal dominant polycystic kidney disease associated with right adrenal adenoma. Refractory hypertension and hypokalemia were the findings that led to this diagnosis. The diagnostic approach included plasma potassium and aldosterone levels, plasma renin activity, captopril test, saline infusion test, NP-59 scintigraphy, computed tomography, and magnetic resonance imaging. Adrenalectomy completely normalized hypertension and hypokalemia in this patient.

Adenoma↗
PubMed1994

Clinicopathologic study of Gore-Tex patch graft in corneoscleral surgery.

Three patients with corneal or scleral defects were treated with Gore-Tex grafting, including one case of corneal ulcer, one case of Mooren's ulcer, and one case of necrotizing scleritis. These eyes remained inflamed after Gore-Tex grafting. The interval between grafting and removal ranged from 2 months to 1 year. Pathologically, none of these grafts showed evident epithelial or fibrous tissue ingrowth of the Gore-Tex graft. Fungal inhabitation was noted in one graft, and fungal endophthalmitis developed 5 months after graft removal and penetrating keratoplasty. Although Gore-Tex offers some advantages, it has disadvantages when it is used without adequate conjunctival covering, including poor epithelialization, poor adhesion between graft and surrounding tissue, and potential infectious route.

Adult↗
PubMed1992

The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase.

X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (approximately 5 A) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the extended x-ray absorption fine structure region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near physiological concentrations (as low as approximately 100 microM). RNA polymerase from Escherichia coli contains two zinc atoms: one tightly bound in the beta' subunit, the subunit that participates in template binding, and the other loosely bound in the beta subunit, the subunit that participates in substrate binding. X-ray absorption studies of these zinc sites in the native protein and of the zinc site in the beta' subunit after removal of the zinc in the beta subunit site by p-(hydroxymercuri)benzenesulfonate (Giedroc, D. P., and Coleman, J. E. (1986) Biochemistry 25, 4969-4978) indicate that both zinc sites have octahedral coordination. The zinc in the beta' subunit site has four sulfur ligands at an average distance of 2.36 +/- 0.02 A and two oxygen (or nitrogen) ligands at an average distance of 2.23 +/- 0.02 A. The beta subunit zinc site has five sulfur ligands at an average distance of 2.38 +/- 0.01 A and one histidine nitrogen ligand at 2.14 +/- 0.02 A. These results are in general agreement with earlier biochemical and spectroscopic studies.

Absorptiometry, Photon↗
PubMed1992

Laparoscopic nephrectomy in a porcine model.

Laparoscopic nephrectomy was performed in 15 male pigs, the procedure was successful in 14. Extraction of the intact kidney through a 5-cm lower abdomen incision was done in 7 animals; complete destruction and evacuation of the kidney was accomplished by a round-knife suction device through a 1-cm port in another 7 pigs. Grossly, the specimen consisted of sausage-like tubular renal tissue and a small amount of tissue debris. Pathology revealed that the glomerular and tubular structures were well preserved, no interstitial hematoma could be found. Four ports were usually used, one 1-cm umbilical camera port, one 0.5-cm port for ureter traction, and two 1-cm working ports along the midclavicular line. All the pigs recovered uneventfully. The average operation time was 3 h 20 min. The application of endo-GIA (United States Surgical Corporation) for renal hilum reduced the operative time to 2 h 20 min. Complications included renal vein tear during endoclip application and cutting in the first case, mild subcutaneous emphysema in 2 cases. This first pig received exploratory laparotomy for the repair and ligation of the renal vein. No more major complications occurred with increasing experience. From this porcine experiment, we conclude that the combination of laparoscopy, a tissue destroyer and an endobag for the entrapment of kidney seem to be a promising technique for clinical nephrectomy.

Animals↗
PubMed1991

Conversion of the E1A Cys4 zinc finger to a nonfunctional His2,Cys2 zinc finger by a single point mutation.

Trans-activation by the adenovirus E1A 289R protein requires a zinc finger defined by Cys-154, Cys-157, Cys-171, and Cys-174. Whereas individually replacing the four cysteine residues with serines resulted in a loss of transactivation, only three of the Cys----Ser mutants (C157S, C171S, and C174S) lost the ability to bind Zn(II). X-ray absorption fine structure analysis revealed that, in the wild-type protein, Zn(II) is coordinated by four cysteine residues whereas in the C154S mutant, Zn(II) is coordinated by two histidines and two cysteines. The mutant protein probably retains, as ligands, two cysteines on the right side of the zinc finger (Cys-171 and Cys-174) and recruits two of the four histidines on the left side (His-149, His-152, His-158, and His-160), despite the presence of Cys-157. This finding may shed light on the general structural requirements of zinc fingers.

Adenoviridae↗
PubMed1988

The 289-amino acid E1A protein of adenovirus binds zinc in a region that is important for trans-activation.

The E1A gene of adenovirus type 5 encodes two major proteins of 289 and 243 amino acid residues, which are identical except that the larger protein has an internal stretch of 46 amino acids required for efficient trans-activation of early viral promoters. This domain contains a consensus zinc finger motif (Cys-Xaa2-Cys-Xaa13-Cys-Xaa2-Cys) in which the cysteine residues serve as postulated ligands. Atomic absorption spectrophotometry applied to bacterially expressed E1A proteins revealed that the 289-amino acid protein binds one zinc ion, whereas the 243-amino acid protein binds no zinc. Replacing individual cysteine residues of the finger with other amino acids destroyed the trans-activating ability of the 289-amino acid protein, even when structurally or functionally conserved amino acids were substituted. These results strongly suggest that the zinc finger of the 46-amino acid domain is intimately linked to the ability of the large E1A protein to stimulate transcription of E1A-inducible promoters. Furthermore, zinc binding to one of the mutant finger proteins suggests either that only a precise finger structure formed by the tetrahedral coordination of zinc to the four consensus ligands is required for trans-activation or, possibly, that one of several neighboring histidine residues in various combinations with three of the consensus cysteine residues normally coordinates zinc. How the zinc finger in E1A might interact with DNA or protein to bring about trans-activation is discussed.

Adenovirus Early Proteins↗
PubMed1982

Role of aromatic residues in the structure-function relationship of alpha-bungarotoxin.

The conformation of alpha-bungarotoxin and its cyanogen bromide cleaved and nitrated derivatives was studied by circular dichroism (CD). Native toxin contains no helices but some beta forms and possibly beta turns. Its ordered conformation is little affected when the peptide bond between Met-27 and Trp-28 is cleaved; however, the CD due to Trp-28 is abolished. The CNBr-cleaved derivative retains its immunoaffinity toward anti-toxin sera but loses its neurotoxicity toward the acetylcholine receptor. On the basis of both CD and fluorescence spectra, Trp-28 is probably stabilized by a short-range interaction with the carboxylate group of Asp-30. The ordered conformation of the toxin is also unaltered when one of the two tyrosine residues, identified as Tyr-54, is nitrated with tetranitromethane. This Tyr(NO2)-54 derivative possesses both immunoaffinity and neurotoxicity. However, the toxin is denatured and loses its activities when the other tyrosine residue, Tyr-24, is also nitrated in 6 M guanidine hydrochloride, even after the denaturant is removed. Spectrophotometric titration of the toxin indicates that Tyr-54 has a normal apparent dissociation constant (pKa = 9.7) and Tyr-24 ionizes at pH above 11.2. Both tyrosine residues are in a polar environment, but Tyr-24 is not readily accessible to reagents and is stabilized by long-range interactions, probably involving Glu-41.

Amino Acid Sequence↗