Initial clinical experience with the Ahmed glaucoma valve implant--correction.
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Choroid Diseases↗
Biomedical subjects
M R Wilson
Publications and source records attributed to M R Wilson.
Initial clinical experience with the Ahmed Glaucoma Valve implant.
PURPOSE: We studied the Ahmed Glaucoma Valve implant, an aqueous shunting device that has a unidirectional valve mechanism designed to prevent postoperative hypotony in eyes with intractable glaucoma. METHODS: In this multicenter, prospective clinical trial, we studied 60 eyes (60 patients) with increased intraocular pressure or glaucoma that had not responded to medical treatment, laser photocoagulation, or previous glaucoma surgery, in which the Ahmed Glaucoma Valve implant was placed to decrease intraocular pressure. RESULTS: Success was defined as intraocular pressure less than 22 mm Hg and greater than 4 mm Hg for two months or longer, intraocular pressure that was lowered by at least 20% from preoperative values (in eyes with preoperative intraocular pressures less than 22 mm Hg), and no additional glaucoma surgery or visually devastating complications. Cumulative probability of success at 12 months was 78%. Eight (13%) of 60 eyes had intraocular pressure less than 5 mm Hg the first postoperative day. Two other eyes had shallow anterior chambers, which required anterior chamber reformation. The major complications associated with the use of the valve were serous choroidal detachments in 13 eyes (22%), blockage of the tube in six eyes (10%), malposition of the tube in four eyes (7%), a suprachoroidal hemorrhage in one eye (2%), and corneal graft rejections in three (19%) of 16 eyes with corneal grafts. CONCLUSIONS: Although the 12-month success with the Ahmed Glaucoma Valve implant is similar to that reported for other drainage devices, the complications associated with overfiltration in the immediate postoperative period appear to be less frequent than with other valved drainage devices. Randomized, prospective studies to compare the Ahmed Glaucoma Valve implant with other drainage devices are needed to make clinical comparisons of the different devices.
Adult↗
Treatment of nude mice with 4-amidinoindan -1- one2 '- amidinohydrazone, a new S-adenosylmethionine decarboxylase inhibitor, delays growth and inhibits metastasis of human melanoma cells.
CGP 48664A (4-amidinoindan-1-one2'-amidinohydrazone) is a novel inhibitor of S-adenosyl-methionine decarboxylase (SAMDC), a key enzyme in the biosynthesis of polyamines, which are themselves essential for proliferation of mammalian cells. Seven different human melanoma cell lines were treated in vitro with CGP 48664A. High, intermediate and low levels of cytostasis were induced in four, one and two melanoma lines, respectively. This cytostasis was reversed by the addition of exogenous spermidine or spermine to the culture medium. The heterogeneous low metastatic (CGP 48664A-resistant) A375P cells and highly metastatic (CGP 48664A-sensitive) A375SM cells were implanted into the subcutis or injected intravenously into nude mice. Systemic daily administration of CGP 48664A significantly reduced the size of cutaneous lesions and the number of lung metastases in mice implanted with A375SM cells. No beneficial effects were found in mice injected with A375P cells. Drug activity was dose dependent, and maximal effects were observed when treatment began in mice with small tumour burdens. The data suggest that CGP 48664A is effective against melanoma metastasis in nude mice and that its activity should be tested in combination with other cytoreductive agents.
Adenosylmethionine Decarboxylase↗
Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts.
Techniques have been developed for extracting, amplifying and directly sequencing mitochondrial DNA (mtDNA) from human hair shafts. The hair shaft is ground in a glass micro-tissue grinder, and the DNA is extracted with organic solvent and purified by filtration. The filtrate subsequently provides the mtDNA template for the PCR. The two hypervariable segments of the mtDNA control region are amplified in four separate reactions. After a purification step to remove unincorporated PCR primers, amplified products are quantitated by capillary electrophoresis and subjected to cycle sequencing. The products are separated and analyzed on an automated DNA sequencer. The mtDNA sequences from the hair shaft match the mtDNA sequences from blood samples taken from the same donor.
Base Sequence↗
Quantitation of polymerase chain reaction products by capillary electrophoresis using laser fluorescence.
In samples where the amount of DNA is limited, the polymerase chain reaction (PCR) can amplify specific regions of the DNA. A quantitative analysis of the PCR product would be desirable to ensure sufficient DNA is available for analysis. In this study, we examine the use of capillary electrophoresis (CE) with laser fluorescence detection for quantitation of PCR products. A coated open tubular capillary was used with a non-gel sieving buffer and a fluorescent intercalating dye to obtain results within 20 minutes. Using an internal standard, peak migration time was below 0.1% relative standard deviation (R.S.D.) with a peak area precision of 3% R.S.D. In comparison to quantitation by hybridization, (i.e., slot blot) and spectrophotometric analysis, capillary electrophoresis shows distinct advantages due to its ability to separate unincorporated primers and PCR byproducts from the targeted PCR product. The results demonstrate that CE can be used to monitor the quality and quantity of the PCR product.
DNA, Viral↗
Composite PAGE: an alternate method for increased separation of amplified short tandem repeat alleles.
Genetic typing of short tandem repeat (STR) loci may require electrophoretic separation techniques which, depending on the locus, can resolve alleles differing in size by only 2, 3 or 4 base pairs (bps). Many such loci can be separated by horizontal, discontinuous polyacrylamide gel electrophoresis (hd-PAGE). However, some loci present particular separation challenges. A composite (or step) gel system consisting of 2 zones, anodal and cathodal, which differ by pore size and ionic strength, has been developed to augment resolution of DNA fragments without a need for additional electrophoretic equipment. Depending on pore size, as well as ionic strength, initial migration can be increased or decreased to effect resolution. When compared with hd-PAGE, this composite gel electrophoretic system can result in enhanced resolution of PCR amplified STR alleles.
Actins↗
Clusterin binds by a multivalent mechanism to the Fc and Fab regions of IgG.
Clusterin was purified from human serum by IgG and monoclonal antibody affinity chromatography. SDS-PAGE and immunoblotting revealed no major differences between clusterin prepared in these two ways. An ELISA method for measuring the binding of clusterin to immunoglobulins was developed. Clusterin purified by IgG affinity chromatography bound to pooled human IgG with a similar affinity (S0.5 5.9 +/- 0.4 micrograms/ml) as clusterin purified by monoclonal antibody chromatography (S0.5 6.1 +/- 0.2 micrograms/ml). The apparent affinity of clusterin for IgG immobilised on ELISA plates increased with increasing concentrations of IgG in the coating solution. Aggregated IgG in solution was a more potent inhibitor of the binding of clusterin to immobilised IgG than was monomer IgG. Clusterin bound to all of the isotypes of human IgG, and to human IgA and IgM, with apparent affinities in the order IgG3 > IgG4 > IgM > IgG1 > IgG2, IgA. Clusterin bound to both the Fab and Fc fragments of human IgG. The clusterin binding site(s) on the Fc do not overlap with those for protein A and Clq.
Antibodies, Monoclonal↗
Structural relationship between the two IgY of the duck, Anas platyrhynchos: molecular genetic evidence.
cDNA clones encoding the H chains of the 7.8S and 5.7S IgY of the White Pekin duck have been isolated and sequenced. The H chain of the 7.8S IgY possesses four C region domains and thus resembles the H chain of chicken IgY with which it shows, in the C region, 54% inferred amino acid sequence identity, and complete conservation of the C region cysteine and tryptophan residues. The H chain of the 5.7S IgY possesses only two C region domains, that are virtually identical to CH1 and CH2 of the 7.8S IgY H chain. Although Southern blot genomic analysis did not resolve whether the two transcripts encoding the H chains of the 7.8S and 5.7S IgY are derived from one or two H chain-encoding genes, the CH 1, 2, 3, and 4 exons are apparently colinear, and no evidence was found for a separate locus in which CH1 and 2 exons were present and CH3 and 4 exons were lacking. The VH domain-encoding sequences of the cDNA for the two IgY H chains showed high similarity in the inferred VH gene (93% nucleotide and 91% inferred amino acid identity) and in the inferred JH segment (89% nucleotide and 93% inferred amino acid identity) but low similarity in the D region (26% nucleotide and 7% inferred amino acid identity). Genomic Southern blot hybridization analysis showed multiple VH-hybridizing sequences represented on up to 20 restriction fragments.
Amino Acid Sequence↗
Alternate splicing pathways of the immunoglobulin heavy chain transcript of a teleost fish, Ictalurus punctatus.
Previously we sequenced a partial cDNA clone encoding the 3' region of the message for the membrane receptor form of the heavy (mu) chain of the channel catfish which indicated that the first transmembrane (TM1) exon is spliced directly to the C mu 3 exon and not into a cryptic site within the CH4 exon, as occurs in other vertebrates. Studies utilizing polymerase chain reaction analysis of mRNA and further analysis of cDNA clones now confirm that the only detectable splicing pattern used in micron production by the channel catfish utilizes this C mu 3----TM1 pathway of pre-mRNA splicing.
Animals↗
Use of phenylpropanolamine to reduce nicotine cessation induced weight gain in rats.
The present study was conducted to determine if phenylpropanolamine (PPA) administered during the first week of nicotine termination could reduce or eliminate the body weight rebound which accompanies nicotine cessation. Sprague-Dawley rats were administered nicotine for 2 weeks after which they received either PPA or saline for 1 week. Control animals received saline during both drug periods. Body weight, food consumption, and water consumption were measured daily before drug, during nicotine and PPA administration, and for 14 days after PPA administration. In contrast to animals receiving saline upon termination of nicotine, animals receiving PPA did not gain weight at an accelerated rate. Termination of PPA did not result in a body weight rebound. To the extent that these results generalize to humans, they suggest that PPA could be used to reduce or eliminate postcessation weight gain in smokers who stop smoking.
Animals↗
RHP is antigenically related to factor H and binds to the globular heads of C1q.
RHP was purified from normal serum by sequential euglobin precipitation, ion exchange chromatography on DEAE-Sephacel and gel filtration using Sephacryl S-300. RHP reacted with anti-Factor H antibodies in ELISA assays and in Western blots, suggesting that it is antigenically related to Factor H. It bound to intact C1q but not to the collagen-like N-terminal half of the molecule. C1q-specific monoclonal antibody BUS-1, which blocks the binding of C1q to immune complexes, did not block the binding of RHP to C1q. This implies that the binding sites on C1q for IgG and RHP do not overlap.
Binding Sites↗
Sequential therapy with chemotherapeutic drugs and liposome-encapsulated muramyl tripeptide: determination of potential interactions between these agents.
Three syngeneic murine tumor models were used to determine potential interactions between chemotherapeutic drugs and the synthetic liposome-encapsulated macrophage activator, muramyl tripeptide phosphatidylethanolamine (MLV-19835). Experiments were designed to maximize any additive toxicity of the simultaneous administration of MLV-19835 on the known myelosuppressive effects of doxorubicin, ifosfamide, and cisplatin. Treatment with these drugs resulted in diminished blood leukocyte counts, altered leukocyte differentials, and decreased hematocrits, but the systemic administration of MLV-19835 produced no additional deleterious effects. Myelosuppression normally observed at 2 weeks following treatment of mice with doxorubicin was prevented by combination treatment with MLV-19835. In addition, there was no interference of the antitumor activity of ifosfamide or doxorubicin against subcutaneous, kidney, and spleen tumors. These studies and the recent demonstration of the biological activity of MLV-19835 in phase II trials of osteosarcoma recommend clinical testing of these combined modalities.
Acetylmuramyl-Alanyl-Isoglutamine↗
Clusterin enhances the formation of insoluble immune complexes.
Clusterin was purified from human serum by sequential affinity chromatography over IgG-, protein A- and Con A-Sepharose. The protein was approximately 70 kDa by SDS/PAGE under nonreducing conditions and was resolved into approximately 35 kDa bands under reducing conditions. The protein reacted with clusterin-specific Mabs in ELISA and in Western blots. Its N-terminal sequences agreed with those published for clusterin. An antiserum specific for clusterin made by the above method detected it in complement membrane attack complexes on rabbit erythrocyte membranes. The interaction of clusterin with IgG was physiologically relevant because it was found to increase the rate of formation of insoluble immune complexes.
Amino Acid Sequence↗
The ocular effects of gases when injected into the anterior chamber of rabbit eyes.
We studied the toxic effects of sulfur hexafluoride and perfluoropropane in comparison with air, balanced salt solution, hyaluronate sodium, and aqueous humor in a rabbit model. Sixty normal pigmented rabbits were studied during a period of 4 weeks. The variables studied were slit-lamp biomicroscopic examination of the anterior segment, intraocular pressure as measured by pneumotonometry, corneal thickness and endothelial cell count as measured by specular microscopy, lens opacity by Scheimpflug photography, and light and transmission electron microscopy. All three gases were more toxic to the cornea and lens than were balanced salt solution, hyaluronate, and aqueous humor. However, 15% perfluoropropane and 50% sulfur hexafluoride were no more toxic to the eye than air was. Therefore, this study suggests that nonexpansile mixtures of perfluoropropane and sulfur hexafluoride may be beneficial and relatively safe in re-forming persistently flat anterior chambers in situations where the use of air is being considered.
Air↗
Immunofluorescent labeling using covalently linked anti-phycoerythrin antibodies and phycoerythrin polymers.
We have developed methods for covalently attaching anti-phycoerythrin (PE) MAbs to other antibodies, and for using PE or polymers of PE in conjunction with these conjugates to rapidly produce specific, high intensity fluorescent labeling of antigens. The performance of these systems was examined on the surface of microspheres and on the cell surface. The noncovalent means by which PE is incorporated into the label complex in this method makes it possible to use crude algal homogenate successfully as a source of PE in immunofluorescence assays. The intensity of labeling achieved using this method is comparable to, or in some cases better than, that obtained using a direct PE conjugate. With the aim of amplifying the intensity of fluorescence obtained, we produced two types of covalently linked complexes. These were an anti-PE MAb, designated PE6, linked to itself (i.e., PE6-PE6), and PE linked to itself (i.e., PE-PE). When used in a two-stage procedure in place of monomeric PE alone, these complexes increased the intensity of fluorescence obtained on the surface of microspheres by more than fourfold. On the cell surface, the performance of this system varied from one antigen to the next but in most cases was restricted to, at best, a 60% increase in the intensity of fluorescence and in many cases only about a 30% increase.
Antibodies, Monoclonal↗
Purification and characterization of infectious Marek's disease virus genomes using pulsed field electrophoresis.
Marek's disease virus (MDV) is an acutely oncogenic avian herpesvirus. The tightly cell-associated in vitro growth characteristics of MDV present unique problems when attempting to purify, analyze, and manipulate MDV genomes. To facilitate molecular characterization of MDV, contour-clamped homogeneous electric fields electrophoresis (CHEF) was used to purify infectious MDV genomes. CHEF techniques were optimized for evaluation of total genome size and alterations in structure which occur during in vitro attenuation of oncogenic MDV. Our results indicated that genomes of attenuated serotype 1 MDV strain JM may contain deletions totaling 15 kbp while high-passage serotype 2 nononcogenic MDV strains SB-1 and 281MI/1 were 5 and 3 kbp larger, respectively, than their low-passage counterparts. Using cell-free CHEF-purified MDV genomes as hybridization probes, we identified a 200-bp deletion in attenuated genomes of the very virulent MDV strain MD11. At present, it is unclear if this 200-bp is related to mutations which lead to loss of oncogenicity or pathogenicity in MD11. This study is the first report which describes procedures for purification of infectious herpesvirus genomes from pulsed-field gels. Our results demonstrate that pulsed-field-purified viral DNA will facilitate molecular characterization of MDV and other cell-associated herpesviruses.
Animals↗
Use of bioerodible polymers impregnated with mitomycin in glaucoma filtration surgery in rabbits.
A prospective, randomized, double-masked, and placebo-controlled study was performed to evaluate the effects of a localized and sustained release of mitomycin on the success of glaucoma filtration surgery in rabbits. A bioerodible polymer was used as the drug carrier. Full-thickness filtration surgeries were performed and data from 22 rabbits were collected. One eye received a polymer impregnated with mitomycin (0.02 mg or 0.06 mg), while the fellow eye received a drug-free polymer. Intraocular pressure, bleb survival, and postoperative complications were investigated. Intraocular pressures remained lower for a longer period of time (P less than 0.004) and filtration blebs lasted longer (P less than 0.05) in experimental eyes than in control eyes. Conjunctivitis and sectorial corneal haze occurred more frequently in eyes treated with the higher dosage mitomycin compared with control eyes. The use of mitomycin-C in a polymer delivery system appeared to promote the success of glaucoma filtration surgery in rabbits. With the lower dosage of mitomycin, clinically significant ocular toxicity was not noted.
Animals↗
The effects of topical mitomycin on glaucoma filtration surgery in rabbits.
A prospective, randomized, double-masked, and placebo controlled study was performed to examine the effects of topically applied mitomycin on the success of full thickness filtration surgery in 23 rabbits. Both eyes of each rabbit had surgery by the same surgeon. After surgery, the experimental eye received a single subconjunctival injection of mitomycin (0.2 mg), and 50 microliters of a 0.2 mg/cc solution of mitomycin topically four time daily. The fellow control eyes were treated with sterile water. The mean intraocular pressure reduction from baseline exhibited by the experimental eyes was greater than that observed for the control eyes. The difference in intraocular pressure reduction between the two groups was statistically significant from post-operative day three through day 21 (p less than .05). With survival analysis, fewer failures were noted in the experimental eyes compared to control eyes for each time period throughout the study (p less than .005). The experimental eyes also demonstrated a longer time to bleb failure although this difference did not achieve statistical significance (p less than .10). An additional five rabbits were treated with the experimental drop in one eye and sterile water in the fellow eye but underwent no surgery. No differences between eyes were noted with regard to intraocular pressure change or anterior segment pathology. These data demonstrated the efficacy of mitomycin in promoting filtration surgical success in rabbits.
Administration, Topical↗