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Biomedical subjects

J Weng

Publications and source records attributed to J Weng.

At least 73 records · Page 4Linked to original sources

The Fas-Fas ligand system and other modulators of apoptosis in the cornea.

PURPOSE: Previous studies have suggested that the disappearance of anterior keratocytes after injury to the overlying epithelium is mediated by apoptosis. The authors examined the expression of the apoptosis-related modulators, Fas (receptor), Fas ligand, Bax, Bcl-2, Bcl-XL, and interleukin-1 beta converting enzyme (ICE) in corneal cells as candidate mediators of this response and tested the effect of Fas receptor-stimulating antibody on corneal stromal fibroblast cells in vitro. METHODS: Reverse-transcription-polymerase chain reaction was used to detect FAS, FAS ligand, Bax, Bcl-2, Bcl-XL, and ICE mRNA expression in primary cultures of human corneal epithelial, stromal fibroblast, and endothelial cells. Immunohistochemistry was applied to detect Fas and Fas ligand proteins in fresh-frozen sections of normal human cornea. The effect of FAS-stimulating monoclonal antibody on first-passage stromal fibroblasts was studied using a DNA fragmentation assay, the live-dead assay with fluorescent microscopy, toluidene blue staining with light microscopy, and electron microscopy. RESULTS: FAS, Fas ligand, Bax, Bcl-2, Bcl-XL, and ICE mRNAs are expressed in all three major cell types of the cornea. Fas protein is expressed in corneal epithelial, keratocyte, and endothelial cells in fresh-frozen human cornea. Fas ligand protein, however, was detected in corneal epithelial and endothelial, but not keratocyte, cells. Fas-stimulating antibody induced first-passage stromal fibroblast cell death with morphologic changes and DNA fragmentation consistent with apoptosis. CONCLUSIONS: The Fas system (Fas and Fas ligand) modulators and final common pathway mediators of apoptosis are expressed in corneal cells. The distribution of Fas (epithelial, keratocyte, and endothelial cells) and Fas ligand (epithelial and endothelial cells) protein expression in fresh-frozen corneal tissue suggests that Fas ligand expressed in corneal epithelial and endothelial cells modulates functions in keratocyte cells and, possibly, autocrine-juxtacrine functions in epithelium and endothelium. The Fas-Fas ligand system is expressed in the cornea and could have important functions in normal corneal physiology and in the pathophysiology of corneal disease, including modulation of keratocyte apoptosis after epithelial injury.

Amino Acid Sequence↗

An ergonomic design and performance evaluation of handy scanners by males.

This paper describes the results of an investigation of the difference in performance, posture used, strain on forearm musculature, and subjective ratings of three handy scanners (models A, B and C). Both models A and B were scanners on the market. Model C was developed for validation. The gripping posture of the three models is distinct both in the anatomical and functional sense. Work with model A requires a thumb-forefinger side grip which induces an ulnar deviation angle; work with model B requires a thumb-finger grip; work with model C requires a thumb-finger-palmar grip. Performance evaluation of scanner models A, B and C on different scanning tasks (with and without stitch) using different resolution modes (100 and 400 dpi), indicated that using the proposed model C resulted in a 13% higher success rate, a 14% shorter completion time, the smallest self-selected working area, least strain on the forearm muscles, and highest subjective ratings among the three. Model C appeared to provide the greatest opportunity for delicate adjustments of posture in response to the activity of the skin receptors, justifying the ergonomic input into the design.

Journal Article↗

Intrinsic factors of apatite influencing its amorphization during plasma-spray coating.

Hydroxyapatite coatings were prepared from differently treated starting powders to investigate the intrinsic factors of apatite influencing its amorphization during plasma-spray coating. The joint analyses of X-ray diffraction and infrared spectra show that the vacancies located on missing hydroxyl sites retard the amorphous/crystalline conversion and enforce retention of the amorphous component; the absorbed water molecules that pre-exist in the starting powder can be incorporated into the vacancies in the hydroxyapatite lattice during plasma-spray coating and compensate for the missing hydroxyls so as to promote the transformation of amorphous into crystalline apatite in the coating process. The more vacancies there are in the apatite structure due to missing hydroxyl sites, the more amorphous the component in the resultant coatings. Moreover, the amorphous phase formed in this way is stable in room conditions.

Biodegradation, Environmental↗

Expression of E6/E7 or SV40 large T antigen-coding oncogenes in human corneal endothelial cells indicates regulated high-proliferative capacity.

PURPOSE: Human corneal endothelial cells are thought to have limited capacity for proliferation. Little is known about the mechanisms that regulate the proliferation of these cells. The authors introduced oncogenes into human corneal endothelial cells to modulate proliferation. In addition, they sought to establish cell lines to facilitate study of human corneal endothelial cells. METHODS: Early-passage human corneal endothelial cells were transduced with disabled retrovirus (pLXSN16E6/E7) coding for the human papilloma virus type 16 transforming oncoproteins E6 and E7. Early-passage cells were also stably transfected by electroporation with the pMTV-D305 plasmid vector, in which SV40 large T antigen (SV40 LTAg) mRNA expression is positively regulated by the mouse mammary tumor virus promoter. Expression of E6/E7 mRNA or SV40 LTAg mRNA in cell lines was monitored with the polymerase chain reaction. SV40 LTAg protein expression was detected by immunocytology and Western blot analysis. RESULTS: Human corneal endothelial cells were efficiently infected with disabled retrovirus coding for E6/E7, and seven strains of cells have continued active proliferation for more than 50 population doublings (PD) (< 8 control PD). E6/E7 mRNA was expressed by each cell strain. E6/E7 transformed cells proliferate rapidly and form a monolayer of cells with a high degree of contact inhibition. Transfection with pMTV-D305 is less efficient, and only a single strain was developed. pMTV-D305-transfected endothelial cells (dexamethasone induced) proliferated at a lower rate than E6/E7-transduced cells or cells transfected with a vector (pSV3neo) in which SV40 LTAg is constitutively regulated. In the absence of dexamethasone, the proliferation of pMTV-D305-transfected cells was even slower, but cells continued to produce SV40 LTAg mRNA and protein. The latter results indicated that SV40 LTAg mRNA continued to be synthesized at significant levels in pMTV-D305-transfected cells in the absence of the inducer dexamethasone. CONCLUSIONS: This study suggests that human corneal endothelial cells have a high capacity for proliferation. Thus, cell division is normally controlled in human corneal endothelial cells by poorly characterized, but efficient, mechanisms. Because the E6 and E7 proteins, as well as the SV40 large T antigen, specifically bind to and interfere with the activity of the retinoblastoma (RB) and p53 tumor suppressor proteins, our results suggest that these proteins have critical roles in regulating the proliferation of human corneal endothelial cells.

Antigens, Polyomavirus Transforming↗

Epidermal growth factor, transforming growth factor alpha, transforming growth factor beta, acidic fibroblast growth factor, basic fibroblast growth factor, and interleukin-1 proteins in the cornea.

The purpose of this study was to determine whether epidermal growth factor (EGF), EGF receptor, transforming growth factor alpha (TGF-alpha), transforming growth factor beta (TGF-beta), acidic fibroblast growth factor (acidic-FGF), basic fibroblast growth factor (basic-FGF), and interleukin-1-alpha (IL-1-alpha) proteins were present in cultures of human corneal cells and/or in sections of human corneal tissue. Immunohistochemistry was performed on human corneal sections. Immunofluorescent cell staining was used to evaluate corneal epithelial, stromal fibroblast, and endothelial cells in primary culture. Basic-FGF production was evaluated in culture cells using immunoprecipitation. EGF, TGF-alpha, TGF-beta-1, and IL-1-alpha were detected by immunohistochemistry in cells in all three layers of the cornea. EGF receptor and acidic FGF were detected by immunohistochemistry in epithelial and endothelial cells, but not in stromal fibroblast cells. Differences in distribution of the growth factors were noted within individual layers of the cornea. EGF and basic-FGF proteins were detected in all three predominant cell types of the cornea using immunocytology. IL-1-alpha protein was detected by immunocytology in corneal epithelial and endothelial cells, but not stromal fibroblasts. Immunoprecipitation confirmed the production of basic-FGF in all three cell types. IL-1-alpha protein detection in the corneal stroma by immunohistology, but not by immunocytology in first passage stromal fibroblasts, suggests that IL-1-alpha may localize to the corneal stroma after production by corneal epithelial and/or endothelial cells.

Adult↗

Effect of epidermal growth factor, hepatocyte growth factor, and keratinocyte growth factor, on proliferation, motility and differentiation of human corneal epithelial cells.

We sought to determine the effects of exogenous epidermal growth factor (EGF), heparin-binding EGF (HB-EGF), transforming growth factor alpha (TGF-alpha), single-chain precursor hepatocyte growth factor (SC-HGF), double-chain mature HGF (DC-HGF), and keratinocyte growth factor (KGF) on proliferation, motility, and differentiation of first passage cultures of human corneal epithelial cells in serum-free chemically defined medium. The effect of EGF, HB-EGF, TGF-alpha, SC-HGF, DC-HGF, KGF or combinations of the growth factors on proliferation was measured by counting cells present after 3 weeks of culture and by immunostaining for the cell-cycle-specific nuclear proliferation antigen Ki-67. The effect of the factors on epithelial cell motility was assessed by morphometric analysis of photographs of cells migrating from confluent islands of cells. The effect of growth factors on differentiation of epithelial cells were determined by immunostaining epithelial cell islands for the keratin K3 and by Western blotting for keratin K3. EGF, alone or in combination with KGF and SC-HGF, significantly stimulated motility of epithelial cells at the periphery of confluent islands of cells and induced an elongated cell morphology. TGF-alpha, HB-EGF and DC-HGF produced motility effects similar to EGF. There was diminished proliferation of the migrating cells in response to EGF, HB-EGF, TGF-alpha or DC-HGF, while non-migrating epithelial cells in the center of confluent islands continued to proliferate in response to the growth factors. EGF, HB-EGF, TGF alpha or DC-HGF inhibited expression of the differentiation-related marker keratin K3 in epithelial cells, both at the edge and at the center of the islands. KGF stimulated proliferation of corneal epithelial cells at low density and in confluent islands of cells. KGF did not affect expression of keratin K3 or migration of epithelial cells. SC-HGF had no effect on corneal epithelial cells. These results indicate that the effects of EGF, HB-EGF, TGF-alpha and DC-HGF on corneal epithelial cell proliferation, motility and differentiation vary from those of KGF and SC-HGF. EGF, HB-EGF, TGF-alpha and DC-HGF induced changes in epithelial cell morphology and motility in cells plated at low cell density or in cells located at the edge of a confluent island. Thus, these effects appear to be dependent on the extent of cell-cell contact. The inhibitory effect of EGF, HB-EGF, TGF-alpha or DC-HGF on corneal epithelial cell differentiation, however, is independent of cell density.(ABSTRACT TRUNCATED AT 400 WORDS)

Blotting, Western↗

Phase and structural changes in hydroxyapatite coatings under heat treatment.

Hydroxyapatite (HA) crystallites of smaller size than those formed during the spraying process are found in HA coatings on titanium as a result of the crystallization of the amorphous phase (approximately 630 degrees C) when the coatings are vacuum-heat-treated in the temperature interval 100-1000 degrees C. As the annealing temperature increases within the 630-1000 degrees C range, the size of the crystallites increases, and at 1000 degrees C reaches the size of those formed during the process of spraying. At the same time, at 800 degrees C and above, HA transforms into other calcium phosphate phases (alpha-tricalcium phosphate, beta-tricalcium phosphate, tetracalcium monoxide diphosphate). These phase transformations lead to the increase of coating roughness.

Biocompatible Materials↗

Mooren-type hepatitis C virus-associated corneal ulceration.

BACKGROUND: Two patients with bilateral Mooren-type ulcers had underlying chronic hepatitis C virus (HCV) infection. Both patients also had chronic, pruritic dermatitis, which in one patient was diagnosed as hidradenitis suppurativa. METHODS: Serum from the first patient and serum, conjunctiva, and liver from the second patient were analyzed for HCV genomic RNA using the reverse transcriptase-polymerase chain reaction. Serum anti-HCV antibodies were monitored with a commercially available second-generation test. Liver and conjunctival biopsies were evaluated histopathologically. RESULTS: Liver biopsy showed severe hepatitis in the first patient, but normal liver tissue in the second. Hepatitis C virus genomic RNA was detected in the serum of both patients. In the first patient, the virus was detected 4 months after completion of interferon alfa-2b treatment for chronic active hepatitis. In the second patient, HCV genomic RNA was detected in serum, but not in conjunctiva or liver tissue. Hepatitis C virus could not be detected in the serum of the second patient after 2 weeks of interferon alfa-2b treatment. Both patients had serum anti-HCV antibodies. In case 1, there was a marked improvement in the corneal disease during and after 6 months of interferon alfa-2b treatment for chronic active hepatitis that paralleled a return of serum liver enzyme levels to the normal range. In the second patient, the corneal disease improved after 6 weeks of interferon alfa-2b treatment, but abruptly worsened when the patient discontinued therapy. The corneal disease improved again after interferon alfa-2b was reinstituted. CONCLUSIONS: Chronic HCV virus infection is associated with Mooren-type peripheral ulcerative keratitis. All patients with Mooren-type ulcers should be tested for evidence of HCV infection in consultation with a liver specialist. Even when improvement is obtained with interferon alfa-2b treatment, however, continued follow-up is important because relapse is common and repeat treatment may be effective.

Adult↗

Effects of beta-endorphin on phytohemagglutinin-induced lymphocyte proliferation and mouse plaque-forming cell response via an opioid receptor mechanism.

The effects of opioid peptides on immune responses were investigated. It was found that beta-endorphin (beta-END) can depress proliferative responses to PHA in rat splenocytes but enhance those in mice, and it could also inhibit the plaque-forming cell (PFC) response to sheep red blood cells when mouse splenocytes immunized in vivo were cultured in vitro with the peptide. The peptide antagonist naloxone was able to reverse beta-END suppression of the PFC response. The data indicate that beta-END suppresses antibody production or secretion via a specific opioid-receptor-mediated mechanism.

Animals↗

Hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), and their receptors in human breast cells and tissues: alternative receptors.

We sought to determine whether the hepatocyte growth factor/scatter factor (HGF/SF)- and keratinocyte growth factor-receptor systems were expressed in normal breast cells, breast carcinoma cell lines, normal breast tissues, and breast cancer tissues. Reverse transcriptase-polymerase chain reaction and hot blotting were used to detect HGF, HGF/SF (met) receptor, KGF, and KGF receptor mRNAs in human mammary epithelial (HME) and stromal (HMS) cells. We also examined breast carcinoma (MDA-MB-157, SCC 38, and SCC 70) and spontaneously immortalized breast epithelial (HMT 3522) cell lines, as well as normal breast and breast carcinoma tissues. PCR products were also confirmed by nucleic acid sequencing. The effects of HGF and KGF, compared to EGF and heparin-binding EGF, on the proliferation of normal human mammary epithelial cells in serum-free defined medium was determined by cell counting. HGF and KGF mRNAs were detected in HMS cells, but not HME cells. KGF receptor mRNA was detected in HME cells, but not HMS cells. HGF/SF receptor mRNA was detected in both HME and HMS cells. mRNAs were also detected in normal breast and breast carcinoma tissues, as well as breast carcinoma and transformed breast epithelial cell lines. Alternative cDNA sequences that are predicted to code for a soluble KGF receptor and a membrane bound, truncated HGF/SF receptor were detected in breast epithelial cells and breast tissues. HGF and KGF maintained viability and stimulated proliferation of HME cells.

Adult↗

Highly sensitive object location in tissue models with linear in-phase and anti-phase multi-element optical arrays in one and two dimensions.

Based upon previous observations of low-frequency photon diffusion waves within highly scattering tissue, this paper explores the "near-field" phenomena of such waves of approximately 10-cm wavelength with 200-MHz phase modulation equipment. Multiple-element source arrays consist of laser diode sources modulated at 180 degrees out of phase with respect to the other sources. The diffusing waves originating from the out-of-phase sources give, in the midplane, an amplitude null and a sharp phase transition. These may be observed in a highly scattering intralipid medium simulating the breast or brain (0.5% intralipid), 3-5 cm from the transmitting laser diodes. In the plane containing the array, there is a high sensitivity for a small volume of a hidden absorber (indocyanine green) deep within a highly scattering medium; 20 pmol in a volume of 70 microliters can be detected. Two-dimensional arrays consisting of four or more elements in two orthogonal planes give sensitivity on both axes similar to the one-dimensional array. Measurements show that in the presence of a light-absorbing object, the amplitude null and the interference plane becomes a curved surface which is deflected toward the heterogeneity. The degree of deflection is related to the volume and the absorption characteristics of the heterogeneity and provides detection of the heterogeneity, and thereby may provide localization information for the detection of small tumors within the human breast, or stroke volumes, aneurysms, and tumors in the human brain.

Brain↗

Amorphous phase and morphological structure of hydroxyapatite plasma coatings.

It is shown that under standard conditions of plasma spraying hydroxyapatite powder reaches the substrate in the form of molten drops. On striking the surface, the molten drop splatters, cools rapidly and solidifies. Consequently, a pancake particle surrounded by tiny spherical or differently shaped particles is formed on the substrate. The solidified particles are partly crystallized. Besides the crystalline hydroxyapatite, they contain some supercooled molten hydroxyapatite, i.e. amorphous hydroxyapatite. Build-up of these particles during coating gives a porous pancake-globular structure. During a brief vacuum heating at 630 degrees C, the amorphous phase is crystallized and completely crystalline hydroxyapatite coatings are obtained.

Crystallization↗

Further studies on the plasma-sprayed amorphous phase in hydroxyapatite coatings and its deamorphization.

Plasma-sprayed hydroxyapatite coatings contain a quite large amount of amorphous phase. Infrared analysis shows that the plasma-sprayed amorphous phase is an oxyapatite and the coating predetermined as a hydroxyapatite is proved to be an oxyhydroxyapatite with a small quantity of hydroxyapatite. Heat treatment promotes the transformation of amorphous oxyapatite into a crystalline hydroxyapatite structure and reduces the dissolution rate of the coatings. Further studies focus on the properties of the amorphous phase, showing that amorphous oxyapatite is more soluble in 0.15 M lactic acid at 25 degrees C and that water molecules can accelerate the amorphous/crystalline transformation of the hydroxyapatite structure during the plasma-spraying process.

Crystallization↗