Applying neural network techniques to plethysmographic pulse shape analysis.
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Adolescent↗
Biomedical subjects
J Weng
Publications and source records attributed to J Weng.
Identification of two rds/peripherin homologs in the chick retina.
PURPOSE: To identify possible homologs of mammalian rds/peripherin in chick photoreceptors. METHODS: An embryonic day-15 chick retinal library was screened by polymerase chain reaction with degenerate oligonucleotide primers derived from conserved segments of the mammalian retinal degeneration slow (rds) mRNA. The resultant amplification products were used to isolate cDNAs, containing complete coding regions. These clones were studied by nucleotide sequence, Northern blot, and in situ hybridization analyses. RESULTS: Two new homologs of rds/peripherin were discovered: crds1 and crds2. The predicted crds1 protein is 78%, and the predicted crds2 protein is 54%, identical to mammalian rds/peripherin. The crds1 mRNA is an abundant 4.4-kb species present in photoreceptors. The crds2 mRNA is of similar size but is much rarer. No homologs of rom1 were identified in our screen. Developmentally, the crds1 mRNAs were first detectable at embryonic day 18. CONCLUSIONS: Crds1 likely represents the chick ortholog of mammalian rds/peripherin, whereas crds2 is a more distant homolog. Both share an elongated C-terminal domain, an unusual feature compared with other members of the rds family.
Amino Acid Sequence↗
Screening of HIV-1 Env glycoproteins for the ability to raise neutralizing antibody using DNA immunization and recombinant vaccinia virus boosting.
HIV-1 envelopes from two series of primary isolates (from Swedish patients 5 and 6), from JR-FL and BaL (prototypic monocyte/macrophage tropic viruses) and from HXB-2 (a prototypic T-cell-line-adapted virus), have been screened for their ability to elicit neutralizing antibody to HIV-1. Rabbits were primed by gene gun inoculation with plasmids expressing secreted monomeric (gp120) and oligomeric (gp140) forms of each Env. After four to six DNA immunizations administered over a 1-year period, rabbits were boosted with 10(8) plaque-forming units of a mixture of seven recombinant vaccinia viruses which express chimeric gp140 Envs (primary clade B sequences in a IIIb-related BH10 backbone). Neutralizing antibodies were assayed against two T-cell-line-adapted viruses (MN and IIIb), two non-syncytium-inducing (NSI) and two syncytium-inducing (SI) primary isolates, and two HIV-1-NL4-3-recombinants with patients 5 or 6 Envs (NL4-3/5A, NL4-3/6C). The DNA priming and recombinant vaccinia virus boosting raised low titers of neutralizing antibody in 10 of 19 rabbits. The highest titers of neutralizing activity (approximately 1:150 for MN) were raised in rabbits DNA primed with Envs from Swedish patients 5. These sera cross neutralized IIIb and MN but did not neutralize the primary isolates or the NL4-3 recombinant with the homologous 5A Env. Sera from rabbits primed with the HXB-2 Env DNA were, for the most part, type-specific for neutralization of IIIb. In one of three assays, sera from rabbits primed with plasmids expressing the JR-FL and BaL had possible low titer neutralizing activity for two NSI, but not two SI, primary isolates. Our results highlight the low immunogenic potential of the HIV-1 Env and demonstrate that different Envs have different potentials to raise low titer neutralizing antibody.
AIDS Vaccines↗
HIV-1 Env glycoproteins from two series of primary isolates: replication phenotype and immunogenicity.
Seven envelope regions from two series of patient isolates have been molecularly cloned and analyzed for replication phenotypes and immunogenicity. Growth potential was analyzed for env sequences substituted into an HIV-1-NL4-3 backbone (NL4-3/env recombinants). Immunogenicity studies were conducted on secreted monomeric (gp120) and oligomeric (gp140) forms of the Envs using Env-expressing plasmid DNAs for immunizations. The env regions of the patient isolates conferred a spectrum of replication kinetics and cytotropisms on the NL4-3/env recombinants. Both patient series included non-syncytium-inducing viruses with no ability to grow on T-cell lines, and highly syncytium inducing viruses which grew well on T-cell lines. These differences in growth potential did not correlate with the ability of the DNA-expressed Envs to raise antibody in rabbits. Rather, the relative immunogenicity of the Envs was patient and form specific. The Envs from patient 5 raised higher titers of antibody than the Envs from patient 6. For each primary Env, the gp120 form of the Env raised higher titers of antibody than the gp140 form. Thus, structural features of Env that affect replication do not necessarily affect the ability to raise antibody.
AIDS Vaccines↗
Fuchs' corneal endothelial cells transduced with the human papilloma virus E6/E7 oncogenes.
We sought to develop corneal endothelial cell cultures with extended lifespan from corneas with Fuchs' dystrophy. Descemet's-endothelial cell explants from histology confirmed recipient corneas of two patients with Fuchs' dystrophy were cultured. After a small number of corneal endothelial cells with irregular, endothelial morphology migrated from the explants onto the culture plate, the cells were transduced with a disabled retrovirus (pLXSN16E6/E7) coding for the human papilloma virus type 16 transforming oncoproteins E6 and E7. Expression of E6/E7 mRNA in the cell cultures with extended lifespan was monitored by RT-PCR. In vitro labeled cellular protein patterns of the Fuchs' E6/E7 cell cultures with extended lifespan were compared with those from normal human corneal endothelial E6/E7 cell cultures with extended lifespan using two-dimensional gel electrophoresis. Two endothelial cell cultures with extended lifespan were derived from Fuchs' corneas. The morphology of the Fuchs' cells expressing E6 and E7 was similar to that of normal corneal endothelial cells expressing E6 and E7. Surprisingly, the rate of proliferation of Fuchs'-derived cells was similar to that of normal endothelial cells transduced with E6/E7. Proliferation of each Fuchs' cell culture with extended lifespan continued for over 30 population doublings. There were limited quantitative differences in the two-dimensional gel electrophoretic protein patterns of the Fuchs'-derived and normal endothelial cell cultures with extended lifespan, respectively. Retroviral integration is dependent on cell proliferation. Thus, cells that migrated from the Fuchs' Descemet's explants were undergoing at least limited in vitro proliferation when the retroviral vector coding for E6/E7 integrated. Fuchs' corneal endothelial cells expressing E6 and E7 had similar proliferation, cellular morphology, and two-dimensional gel protein electrophoretic patterns to normal corneal endothelial cells expressing the same oncoproteins.
Cell Division↗
Formation and characteristics of the apatite layer on plasma-sprayed hydroxyapatite coatings in simulated body fluid.
Plasma-sprayed hydroxyapatite (HA) coatings were incubated in simulated body fluids (SBFs) for different periods of time to investigate the nucleation and growth of apatite on their surface. The layer that formed was recognized as having similarities to bone apatite because it is poorly crystallized, non-stoichiometric or calcium deficient, and contains carbonate and magnesium. Scanning electron microscopy (SEM) and infrared spectroscopy (IR) were employed to investigate the morphological changes of the coating surface and the structure of the grown layer respectively. In the first few hours, calcium and phosphate ions dissolved from the coatings so as to increase their local supersaturation to a higher degree, thereafter followed by the nucleation and growth of apatite. The nucleation occurred firstly on the recessed regions, inside pores and cracks where the higher supersaturation was readily maintained. Only after 24 h incubation was a complete layer formed on the surface of the coating. There is no obvious interface between the grown layer and the underlying coating. Heat treatment in the air made the apatite transform into biphasic calcium phosphate of HA and tricalcium phosphate, with a blue colour because of trace manganese ions. The heat-treated HA coating showed no dissolution by SEM observation. This resulted in no precipitation on the surface. When SBF was used with two-fold higher ion concentrations, the apatite layer formed slowly in 72 h without dissolution of the coating surface. This may mean that the microenvironment with a sufficiently high degree of supersaturation of calcium and phosphate ions is crucial for apatite to nucleate and grow in SBF, while the HA crystalline structure is not critical in the nucleation process, as expected.
Apatites↗
Characterization of hydroxyapatite film with mixed interface by Ar+ ion beam enhanced deposition.
Ar+ ion beam enhanced deposition (IBED) was used to produce a hydroxyapatite (HA) film on polished titanium substrates. In this study, the HA ceramic target was sputtered by an argon-ion beam with an energy of 1.5 KeV, and the sputtered film was intermittently bombarded by energetic argon-ions at 60 KeV. An effective Ca-Ti mixed layer produced by the energetic argon-ion bombardment was confirmed by using Auger electron spectroscopy. The characteristics of the deposited films were evaluated by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction (XRD) and scanning electron microscopy (SEM) analyses. XRD analysis revealed that the as-deposited film was amorphous, and a hydroxyapatite-type structure was obtained from the post-heat treatment of the deposited films. SEM observations showed that no distinct difference in surface morphology was found between the as-deposited and heat-treated samples for Ar+ IBED films, suggesting a strongly bonded HA film on the titanium substrate. In comparison with the HA target, some chemistry alterations were brought about in the deposited films, such as the incorporation of CO3, the loss of the OH groups and some distortion of the phosphate lattice.
Argon↗
Learning-based ventricle detection from cardiac MR and CT images.
The objective of this work is to investigate the issue of automatically detecting regions of interest (ROI's) in medical images. It is assumed that the regions to be detected can be roughly segmented by a threshold based on a likelihood measure of the ROI. First, an analysis of the global histogram is used to compute a preliminary threshold that is likely near the optimal one. The histogram analysis is motivated by the analytical result of a bell image intensity model proposed in this work. Then, the preliminary threshold is used to segment the input image, resulting in an attention map, which contains an attention region that approximates the ROI as well as many spurious ones. Due to the nonoptimality of the preliminary threshold, it can happen that the attention region contains a part of, or more regions than, the ROI. Learning takes place in two stages: 1) learning for automatic selection of the preliminary threshold value and 2) learning for automatically selecting the ROI from the attention map while dynamically tuning the threshold according to the learned-likelihood function. Experiments have been conducted to approximately locate the endocardium boundaries of the left and right ventricles from gradient-echo magnetic resonance (MR) images. Cardiac computed tomography (CT) images have also been used for testing. The boundary of the segmented region provided by this algorithm is not very accurate and is meant to be used for further fine tuning based on other application-specific measures.
Algorithms↗
[Effects of PGMS on the functions of human platelets and endothelial cells].
OBJECTIVE: To study the effects of Propylene glycol mannte sulfate (PGMS), a heparin-like substance on the functions of platelets and endothelial cells. METHODS: Using 125I labeled monoclonal antibody to measure the expression of GMP-140 on the surface of human platelets; and using naphthol blue black stain method to study the growth of human umbilical vein endothelial cells (HUVEC) and human umbilical vein smooth muscle cells (HUVSMG). RESULTS: PGMS significantly inhibited thrombin stimulated expression of GMP-140 on the surface of human platelets. IC50 was 127 micrograms/ml. Indomethacin also inhibited GMP-140 expression, but its effect was weaker than that of PGMS with the IC50 of 502 micrograms/ml. PGMS did not affect the growth of HUVSMC. On the contrary, it significantly promoted the growth of HUVEC at very low concentration (1.6 micrograms/ml). CONCLUSIONS: PGMS showes anticoagulative effects on both platelets and endothelial cells and it might well be developed as a new antithrombotic and anticoagulateve agent.
Anticoagulants↗
[Analysis of the pyridinoline and deoxypyridinoline in urine by reversed-phase high performance liquid chromatography (RP-HPLC)].
A method for the simultaneous and rapid determination of pyridinoline and deoxypyridinoline in urine using RP-HPLC has been established. After hydrolysis for 18 hours at 107 degrees C in 6 mol/L HCl, the urine sample was prefractionated by partition chromatography using a column packed with cellulose CF1. The appropriate fractions were freeze-dried. The samples were analyzed by HPLC with a 5 microm Spherisorb C18 column, a mobile phase of MeOH/H2O containing 0.1% heptafluorobutyric acid at a rate of 1.2 mL/min and fluorescence detector monitoring at lambda(ex) 290 nm and lambda(em) 400 nm. The limit of a detection for PYD was 10 nmol/L and DPD 7 nmol/L. The recovery of PYD was 91.5% and that of DPD was 106.1%. Within day CV for the two compounds were 1.39% and 0.16% respectively and day-to-day CV 3.71% and 1.32% respectively.
Amino Acids↗
IL-1 upregulates keratinocyte growth factor and hepatocyte growth factor mRNA and protein production by cultured stromal fibroblast cells: interleukin-1 beta expression in the cornea.
PURPOSE: To determine whether interleukin 1 beta (IL-1 beta) messenger RNA (mRNA) and protein were expressed in corneal cells and to examine the effects of IL-1 alpha and IL-1 beta on the expression of hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) mRNAs and proteins in corneal stromal fibroblasts. METHODS: IL-1 beta mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). IL-1 beta protein was detected by immunohistologic tests. Changes in the expression of HGF and KGF mRNAs and proteins in response to stimulation of cultured corneal stromal fibroblasts with IL-1 alpha and IL-1 beta were monitored by Northern and Western blotting, respectively. RESULTS: IL-1 beta mRNA is expressed in human primary cultured corneal epithelial, stromal fibroblast, and endothelial cells. IL-1 beta protein was detected in epithelium and endothelium in fresh frozen human and rabbit corneal tissue. Little, if any, IL-1 beta was detected in the unwounded corneal stroma. IL-1 alpha and IL-1 beta at 10 ng/ml upregulated the levels of HGF and KGF mRNAs and proteins in cultured human corneal fibroblasts. CONCLUSIONS: IL-1 alpha and IL-1 beta may serve as key modulators in an epithelial-stromal regulatory loop in the cornea. These data and previously published observations support the hypothesis that corneal epithelial wounding releases IL-1 alpha and IL-1 beta from epithelial cells; these cytokines in turn upregulate HGF and KGF mRNA and protein levels in keratocytes, and HGF and KGF released by the keratocytes modulate healing of the wounded corneal epithelial cells by regulating proliferation, motility, and differentiation.
Animals↗
Hepatocyte growth factor, keratinocyte growth factor, and other growth factor-receptor systems in the lens.
PURPOSE: To examine the expression and function of hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), epidermal growth factor (EGF) and other growth factor-cytokine-receptor systems in lens epithelial cells. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis were used to examine the expression of messenger RNAs in primary cultured rabbit and human lens cells and in ex vivo rabbit lens tissue. Protein expression and the effect of HGF and KGF on crystallin expression in lens epithelial cells were evaluated by immunoprecipitation and Western blot analysis. The effect of exogenous HGF, KGF, and EGF and of the coculture of lens epithelial cells with corneal endothelial cells on the proliferation of rabbit lens cells in a Transwell system was determined by cell counting. RESULTS: Messenger RNAs and proteins of HGF and KGF were expressed in primary rabbit lens epithelial cells and in ex vivo rabbit lens epithelial tissue. Human lens cells also expressed the mRNAs. Other growth factors and receptor messenger RNAs were also expressed. Hepatocyte and keratinocyte growth factors, and coculture with corneal endothelial cells stimulated proliferation of rabbit lens epithelial cells. In first-passage rabbit lens cells, HGF, KGF, and EGF increased the expression of alpha and beta crystallins. CONCLUSIONS: Hepatocyte and keratinocyte growth factor-receptor systems are expressed in lens cells. HGF and KGF are not expressed in epithelial cells in such tissues as skin, cornea, and lacrimal gland in which fibroblastic and epithelial cells interact in the formation of an organ. Expression of these growth factors in the lens may have evolved because the lens cells are relatively isolated within the anterior chamber of the eye. Our results suggest, however, that growth factors released by the corneal endothelium also could modulate lens functions (aquecrine interactions).
Animals↗
Immune responses to inactivated vaccine in people naturally infected with hantaviruses.
An inactivated Hantaan virus vaccine for hemorrhagic fever with renal syndrome (HFRS) was given by injection to 15 people who were naturally infected with either Hantaan or Seoul viruses. Immunofluorescent antibody (IFA), reversed passive hemagglutination inhibition (RPHI), hemagglutination inhibition (HI), and neutralization antibody (NA) assays were used to measure the antibody titers of the vaccinated people before and after three doses of vaccine. The results indicated that IFA and RPHI antibody titers were boosted significantly (P < 0.05) after the vaccination. Either Hantaan or Seoul virus could induce two-way cross-reactive neutralization antibody responses in humans. After HTNV vaccine immunization, the NA titers of people with natural infection increased significantly (P < 0.05) to both Hantaan and Seoul viruses, while the relative dominance between these two type responses was still similar to that of natural infection. It is worthwhile to studying the procedure further to inoculate two different virus vaccines for improving the cross-protective effect.
Adolescent↗
Integrity and thermal decomposition of apatite in coatings influenced by underlying titanium during plasma spraying and post-heat-treatment.
The integrity and thermal decomposition of calcium apatite are influenced by the underlying titanium during plasma-spraying deposition, especially at the apatite/titanium interface. The destruction of apatite at the interface is governed by substrate temperature, titanium catalysis, and its reaction with titanium dioxide produced from oxidation of titanium in the plasma gas. The apatite in the outer layer of coatings is affected mainly by the substrate temperature and can keep its integrity with a suitable plasma-spraying procedure to minimize the increase of substrate temperature. The heat treatment of the coatings in vacuum results in the decomposition of apatite to alpha-tricalcium phosphate (alpha-TCP) and tetracalcium phosphate monoxide (TCPM) with the increase of intensity approaching the interface, which roughens the surface of the coatings. In the air-heat treatment, oxidation of titanium produces a thickened, dense rutile layer at the interface which prevents titanium atoms from diffusing into the coatings and inhibits the titanium-catalyzed decomposition of apatite. The apatite adjacent to the rutile layer reacts moderately with rutile to produce calcium titanate (CaTiO3), alpha- and beta-TCP, while the apatite in the outer layer, separated from the rutile layer, maintains its integrity without decomposition even in a prolonged air-heat treatment. The retention of apatite integrity leads to a decreased surface roughness of the coating.
Biocompatible Materials↗
Epithelial injury induces keratocyte apoptosis: hypothesized role for the interleukin-1 system in the modulation of corneal tissue organization and wound healing.
The purpose of this study was to determine the role of programmed cell death (apoptosis) in the disappearance of keratocytes beneath an epithelial debridement wound in the cornea and to investigate a potential role of interleukin-1 (IL-1) in induction of apoptosis in stromal fibroblasts in vitro and keratocytes in vivo. Keratocyte and stromal fibroblast cell morphology was examined in wounded and unwounded mouse corneas using transmission electron microscopy. Nuclear DNA fragmentation was detected with the TUNEL assay for 3'-hydroxyl DNA ends. The effect of IL-1 on keratocytes in vivo was determined by microinjection of IL-1 alpha into the central corneal stroma via a limbal entry site. The in vitro effects of interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) were determined with primary cultures of human corneal stromal and dermal fibroblasts. Cell shrinkage, blebbing with formation of membrane bound bodies, condensation and fragmentation of the chromatin, and DNA fragmentation, consistent with apoptosis were detected in anterior stromal keratocytes after epithelial scrape wounds. Thus, disappearance of keratocytes from the underlying stroma following epithelial debridement is mediated by apoptosis. Microinjection of IL-1 alpha into the central stroma of the mouse cornea caused a redistribution of keratocytes in the stroma via apoptosis and, possibly, negative chemotaxis. IL-1 alpha and IL-1 beta induced apoptosis in corneal stromal and dermal fibroblasts in vitro. The epithelial/endothelial-stromal IL-1 system may mediate corneal tissue organization and responses to mechanical- and pathogen-induced injury through induction of keratocyte apoptosis. Keratocyte apoptosis is likely an initiating event in wound healing following corneal surgery. We hypothesize that derangement's in this system may have a role in the pathogenesis of keratoconus and other diseases of the cornea.
Animals↗
Water vapour-treated hydroxyapatite coatings after plasma spraying and their characteristics.
A novel way to enhance the ability of hydroxyapatite (HA) coatings in resisting degradation was revealed. The as-received plasma sprayed HA coatings were kept in water vapour at 125 degrees C, with a pressure of 0.15 MPa for 6 h; most of the amorphous phase in the coating was converted into crystalline HA and enhanced the crystallinity significantly. Meanwhile, the alpha-tricalcium phosphate, tetracalcium phosphate and CaO which decomposed from HA during plasma spraying were also transformed into crystalline HA. The dissolution experiment in distilled water at room temperature showed that the post-water vapour-treated coatings were more stable than post-heat-treated ones. The average interfacial tensile bond strength between HA and substrate before and after water vapour treatment was 45.0 and 39.1 MPa, respectively.
Biocompatible Materials↗
Inactivation of brainstem motor nuclei blocks expression but not acquisition of the rabbit's classically conditioned eyeblink response.
Rabbits were eyeblink conditioned while their accessory abducens nucleus (ACC), facial nucleus (FN), and surrounding reticular formation (RF) were temporarily inactivated with microinjections of muscimol to determine whether these structures are critically involved in acquisition of the conditioned eyeblink response (CR). Rabbits performed no CRs or unconditioned responses (URs) during inactivation training. Training was continued without inactivation and rabbits performed the CR at asymptotic levels from the start of training without inactivation. They had fully learned the CR while their ACC, FN, and RF were inactivated, despite performing no CRs or URs at all during inactivation. These results rule out any critical role for neurons within the ACC, FN, or surrounding RF in acquisition of the classically conditioned eyeblink response.
Abducens Nerve↗
Hepatocyte growth factor and hepatocyte growth factor receptor in the lacrimal gland, tears, and cornea.
PURPOSE: The purpose of this study was to characterize the expression of hepatocyte growth factor (HGF) and HGF receptor proteins in lacrimal gland, tears, and cornea. METHODS: The reverse transcription-polymerase chain reaction method was used to detect HGF and HGF receptor messenger RNA in human lacrimal gland tissue. HGF and HGF (c-met) receptor monoclonal antibody specificity was demonstrated with fluorescent antibody sorting of cells engineered to express HGF or HGF receptor compared with control cell lines, enzyme-linked immunoadsorbent assay (ELISA), immunoprecipitation, and immunohistology with preabsorption. Immunohistochemistry was applied to study the distribution of HGF and HGF receptor expression in rabbit lacrimal gland tissue and in wounded and unwounded rabbit cornea. An ELISA was used to detect HGF in pooled samples of human tears and individual aliquots of tears collected from patients 1 day after anterior segment surgery. RESULTS: Amplification products of the expected size for HGF and HGF receptor mRNAs were detected in lacrimal tissue and were confirmed to be specific by hot blotting and nucleic acid sequencing. Hepatocyte growth factor protein was detected in interalveolar and interlobular connective tissue cells adjacent to glandular alveolar (acinar) cells and associated with the cells lining the interlobular ducts. Hepatocyte growth factor receptor protein was expressed in the glandular alveolar and interlobular ductal cells in the lacrimal gland and all three cell types of the cornea. It was detected in keratocyte and endothelial cells, and expression was increased in keratocytes after epithelial wounding. Hepatocyte growth factor was not present in corneal epithelial cells, but in the unwounded cornea a strong signal was associated with the epithelial cell surface. It was detected by ELISA in pooled normal tears at levels 186 to 290 pg/ml and in individual postoperative tear samples at 453 to 619 pg/ml. In some tear samples, HGF levels were below the sensitivity of the assay (97.5 pg/ml). CONCLUSIONS: The distribution of HGF receptor protein expression in the lacrimal gland suggests that HGF secreted by interalveolar connective tissue cells traverses the acinar cells and modulates functions in acinar and ductal epithelial cells. Hepatocyte growth factor likely collects within the interlobular ducts and becomes a component in normal tears. Thus, lacrimal gland HGF probably modulates corneal epithelial cell proliferation, motility, and differentiation. Its expression in keratocytes is upregulated after corneal epithelial wounding and probably contributes to the epithelial wound healing process.
Animals↗