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The long-term outcome of glaucoma filtration surgery.

PURPOSE: To determine the long-term outcome of glaucoma filtration surgery in preserving vision. Visual loss from progressive glaucomatous damage and from complications of surgery, both short and long term, were included. METHODS: A retrospective, community-based, longitudinal study of residents of Olmsted County, Minnesota, who were newly diagnosed with open-angle glaucoma between 1965 and 1980 and underwent filtration surgery in these or subsequent years through 1998. Intraocular pressure (IOP), visual acuity, visual fields, and progression to legal blindness were monitored. Kaplan-Meier analysis was used to determine the cumulative probabilities of changes in these parameters. RESULTS: 73 eyes of 49 patients underwent conventional filtration surgery. Analysis of the first eye having surgery revealed a mean preoperative IOP of 27.6 +/- 8.5 mm dropping to 16.7 +/- 5.6 mm at year one, and remaining in this range throughout follow-up (14.7 +/- 3.0 mm at 10 years; with or without use of medications). The probability of progression to blindness was 46% at 10 years after surgery, as calculated by Kaplan-Meier analysis. Eyes going blind had a postoperative IOP equal to or lower than those not becoming blind (14.0 +/- 4.4 vs. 15.4 +/- 3.0 at postoperative year 10). Eyes going blind had more advanced field loss at the time of surgery, with scotomas above and below the horizontal axis, than eyes not going blind, which had scotomas in only one hemifield. Three patients developed late bleb leaks; two patients developed endophthalmitis. The probability of undergoing cataract surgery was 37% by 10 years postoperatively, which did not differ significantly from the cohort of patients not undergoing surgery at a comparable time point. CONCLUSIONS: Filtration surgery was associated with a 54% probability of preservation of vision from progression to legal blindness at 10 years after surgery. Patients becoming blind had more advanced field loss at the time of surgery; IOP was similar between those going blind and those retaining vision.

Aged↗

Nephrin localizes at the podocyte filtration slit area and is characteristically spliced in the human kidney.

Defects in the newly reported gene NPHS1 in chromosome 19 cause the massive proteinuria of Finnish type congenital nephrotic syndrome (CNF). Together with its gene product, nephrin, NPHS1 is providing new understanding of the pathophysiological mechanisms of glomerular filtration. Here we show the characteristic splicing of NPHS1 mRNA in the normal and CNF kidneys and localize nephrin exclusively in the glomerulus and to the filtration slit area by light and immunoelectron microscopy. These results indicate that nephrin is a new protein of the interpodocyte filtration slit area with a profound role in the pathophysiology of the filtration barrier.

Amino Acid Sequence↗

Microplate gel-filtration method for radioligand-binding assays.

A thin-layer gel-filtration chromatographic method has been developed in a 96-well format to separate free and protein-bound ligand in radioligand-binding assays. The mobile phase in the gel-filtration plate is removed via quick centrifugation before samples are applied. Protein-bound ligand is recovered via centrifugation into another 96-well plate for radioactivity measurements. The method exhibits excellent recovery of protein-ligand complexes and less opportunity for dissociation of the complexes since it eliminates major dilution effects from the mobile phase of a column and from elution steps in conventional gel-filtration chromatography. It offers other advantages: simple, rapid, inexpensive, quantitative, and able to handle a large number of samples as required in drug discovery and clinical settings. This microplate gel-filtration method was optimized in studies of receptor-ligand interactions using estrogen receptors as examples and can be used in other radioligand-binding assays.

Binding, Competitive↗

Lubrication of the human ankle joint in walking with the synovial fluid filtrated by the cartilage with the surface zone worn out: steady pure sliding motion.

A mixture model of synovial fluid filtration by cartilage in the human ankle joint during walking is presented for steady sliding motion of the articular surfaces. In the paper the cartilage surface zone is assumed worn out. The same model has been recently applied to the squeeze-film problem for the human hip joint loaded by the body weight during standing (Hlavácek, Journal of Biomechanics 26, 1145-1150, 1151-1160, 1993; Hlavácek and Novák, Journal of Biomechanics 28, 1193-1198, 1199-1205, 1995). The linear biphasic model for cartilage (elastic porous matrix + ideal fluid) due to Prof. V. C. Mow and his co-workers and the biphasic model for synovial fluid (viscous fluid + ideal fluid), as used in the above-mentioned squeeze-film problem, are applied. For the physiologic parameters of the ankle joint during walking, a continuous synovial fluid film about 1 microm thick is maintained under steady entraining motion according to the classical model without the fluid transport across the articular surface. This is not the case in the filtration model with the cartilage surface zones worn out. On the contrary, this filtration model indicates that synovial fluid is intensively filtrated by such cartilage, so that no continuous fluid film is maintained and a synovial gel layer, about 10(-8) m thick, develops over the majority of the contact. Thus, if the cartilage surface zones are worn out, boundary lubrication should prevail in the ankle joint under steady sliding motion for the mean values of loading and the sliding velocity encountered in walking cycle.

Ankle Joint↗

Analysis of lipopolysaccharide (LPS)-binding characteristics of serum components using gel filtration of FITC-labeled LPS.

Lipopolysaccharide (LPS)-binding components in serum play an important role in modifying LPS toxicity. We analyzed the binding characteristics of LPS in the presence of serum using gel filtration of FITC-labeled LPS (FITC-LPS) with on line detection of optical density and fluorescence. FITC-LPS separately behaves as an aggregate resulting in a low, dequenched, fluorescence. Binding of single LPS molecules, segregated from the aggregate, to serum components results in an increase in the fluorescence due to dequenching, and a comigration of fluorescence and optical density signals using gel filtration. This method, in combination with the use of specific antibodies inducing additional shifts, demonstrated that in serum high-density lipoproteins (HDL), albumin and low-density lipoproteins (LDL) were able to monomerize LPS. An ELISA on collected fractions of the gel filtration revealed binding of the recently identified LPS-binding protein, serum amyloid P component (SAP), to the high molecular weight LPS aggregate. In serum, binding of soluble CD14 (sCD14) and LPS-binding protein (LBP) to LPS could not be detected. However, this was probably due to an overshadowing effect of albumin, as an extra addition of recombinant sCD14 to serum clearly monomerized FITC-LPS. Biosensor technology revealed that, of all LPS-binding components tested, only SAP clearly bound to the LPS-coated sensor chip. These results show that gel filtration of FITC-LPS is a quick and reliable method to study the binding characteristics of LPS-binding components.

Acute-Phase Proteins↗

The effect of total vitrectomy on filtration surgery in the aphakic eye.

The influence of total vitrectomy on the success rate of filtration surgery was investigated, with 47 operations (6 reoperations) being performed on 41 patients. At six months postoperatively, successful control (IOP less than 21 mm Hg) was achieved in 32 of 41 patients (78%). Twenty-four patients had filtration blebs present. Total vitrectomy favorably influences the result of filtration surgery. However, the morbidity rate (two retinal detachments and two cases of cystoid macular edema) is higher than in standard filtration procedures.

Adult↗

Glaucoma filtration surgery in rabbits using bioerodible polymers and 5-fluorouracil.

A prospective, randomized, double-masked, and placebo-controlled study was performed to examine the effect of a localized and sustained delivery of 5-fluorouracil (5-FU) on the success of glaucoma filtration surgery in 18 rabbits. A bioerodible polyanhydride composed of bis (p-carboxyphenoxy) hexane (PCPH) and sebacic acid (SA) was used as the drug carrier. The polymer and 5-FU (10% by weight) were molded into 4-mm long cylinders by a 15-gauge needle. These implants, with and without the therapeutic agent, were placed at the site of filtration surgery intraoperatively. The results showed that intraocular pressures (IOPs) were lower in the experimental eyes during the second postoperative week, but eventually both experimental and control eyes returned to preoperative levels. Filtration blebs lasted longer in experimental eyes and bleb failure occurred before IOP failure in both experimental and control eyes. Implant disappearance occurred after IOP and bleb failure. Experimental eyes had more postoperative complications than control eyes. Eventually, the filtration surgery failed in both the experimental and control rabbit eyes.

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↗

Filtration capture immunoassay for bacteria: optimization and potential for urinalysis.

A novel assay utilizing immuno-labeling, filtration, and electrochemistry for the rapid detection of bacteria has been optimized for the detection of Escherichia coli O157:H7. Bacteria were specifically labeled with alkaline phosphatase conjugated polyclonal antibodies and captured on a polycarbonate track-etched membrane filter (0.2 microm pore size). The filter was then placed directly against a glassy carbon electrode, incubated with enzyme substrate, and the product detected by square wave voltammetry. The high speed and capture efficiency of membrane filtration and inherent sensitivity of electrochemical detection produced a 25-min assay with a detection limit of 5 x 10(3) E. coli O157:H7 per ml using a filtration volume of 100 microl (i.e. 500 cells filtered). The labeling, filtration, and electrochemical steps were optimized, and the assay performance using electrochemical and colorimetric detection methods was compared. The assay was used to detect E. coli O157:H7 that was spiked into filter-sterilized urine at clinically relevant concentrations.

Bacteriological Techniques↗

Determination of meropenem in plasma and filtrate-dialysate from patients under continuous veno-venous haemodiafiltration by SPE-LC.

Meropenem, a carbapenem antibiotic displaying a broad spectrum of antibacterial activity, is administered in Medical Intensive Care Unit to critically ill patients undergoing continuous veno-venous haemodiafiltration (CVVHDF). However, there are limited data available to substantial rational dosing decisions in this condition. In an attempt to refine our knowledge and propose a rationally designed dosage regimen, we have developed a HPLC method to determine meropenem after solid-phase extraction (SPE) of plasma and dialysate fluids obtained from patients under CVVHDF. The assay comprises the simultaneous measurement of meropenem's open-ring metabolite UK-1a, whose fate has never been studied in CVVHDF patients. The clean-up procedure involved a SPE on C18 cartridge. Matrix components were eliminated with phosphate buffer pH 7.4 followed by 15:85 MeOH-phosphate buffer pH 7.4. Meropenem and UK-1a were subsequently desorbed with MeOH. The eluates were evaporated under nitrogen at room temperature (RT) and reconstituted in phosphate buffer pH 7.4. Separation was performed at RT on a Nucleosil 100-5 microm C18 AB cartridge column (125 x 4 mm I.D.) equipped with a guard column (8 x 4 mm I.D.) with UV-DAD detection set at 208 nm. The mobile phase was 1 ml min(-1), using a step-wise gradient elution program: %MeOH/0.005 M tetrabutylammonium chloride pH 7.4; 10/90-50/50 in 27 min. Over the range of 5-100 microg ml(-1), the regression coefficient of the calibration curves (plasma and dialysate) were >0.998. The absolute extraction recoveries of meropenem and UK-1a in plasma and filtrate-dialysate were stable and ranged from 88-93 to 72-77% for meropenem, and from 95-104 to 75-82% for UK-1a. In plasma and filtrate-dialysate, respectively, the mean intra-assay precision was 4.1 and 2.6% for meropenem and 4.2 and 3.7% for UK-1a. The inter-assay variability was 2.8 and 3.6% for meropenem and 2.3 and 2.8% for UK-1a. The accuracy was satisfactory for both meropenem and UK-1a with deviation never exceeding 9.0% of the nominal concentrations. The stability of meropenem, studied in biological samples left at RT and at +4 degrees C, was satisfactory with < 5% degradation after 1.5 h in blood but reached 22% in filtrate-dialysate samples stored at RT for 8 h, precluding accurate measurements of meropenem excreted unchanged in the filtrate-dialysate left at RT during the CVVHDF procedure. The method reported here enables accurate measurements of meropenem in critically ill patients under CVVHDF, making dosage individualisation possible in such patients. The levels of the metabolite UK-1a encountered in this population of patients were higher than those observed in healthy volunteers but was similar to those observed in patients with renal impairment under hemodialysis.

Anti-Bacterial Agents↗

A mechanistic model of plasma filtration.

A model describing the sieving and transmembrane pressure behavior of plasma filtration is developed and numerically simulated. The model assumes a mechanistic criteria for particle passage through a membrane with cylindrical pores. The initial pore diameter distribution and porosity are assumed to be known. Model inputs include the particle diameter distribution, concentration and total flow rate of the permeate plasma solution. Outputs of the model include transmembrane pressure, the time-averaged sieving coefficients, and size distributions of the deposited particles and accumulated filtrate particles. Optimal filtration is characterized by high, stable sieving coefficients for desired particles, high retention of larger particles and relatively small increases in transmembrane pressure. These characteristics are realized for membranes with mean pore diameters equal to or slightly larger than mean permeate particle diameters. Simulations demonstrate that the incorporation of membrane properties into models of plasma filtration is both significant and readily possible.

Biophysical Phenomena↗

Metal accumulation, filtration and O(2) uptake rates in the mussel Perna perna (Mollusca: Bivalvia) exposed to Hg(2+), Cu(2+) and Zn(2+).

Tissue metal concentrations, filtration and oxygen uptake rates were investigated for Perna perna (Bivalvia: Mollusca) during exposure to Hg(2+), Cu(2+) and Zn(2+) (50 microg/l for 24 days, and 24 days recovery with no metal). Hg and Cu tissue levels increased with exposure time, reaching maximum levels after 24 days (87.5 microg Hg/g dry mass and 45 microg Cu/g dry mass, respectively). Zn levels peaked after 4 days exposure (to 233 microg Zn/g dry mass) and stabilized thereafter. Accumulated metal was rapidly lost from tissues when mussels were returned to uncontaminated seawater, suggesting that tissue concentration data may be of limited use in biomonitoring situations where environmental metals fluctuate to low levels. Filtration rates fell below control rates during Hg(2+) exposure, and became elevated again during the recovery period. Cu(2+) and Zn(2+) exposure had little effect on filtration, but suppressed oxygen uptake. During recovery, oxygen uptake of Cu(2+) and Zn(2+) exposed mussels was elevated above the controls. Filtration and oxygen uptake rates were not correlated, but rather responded in different ways to metal pollution. While these physiological responses of P. perna may be of limited use in biomonitoring, they could indicate how populations may respond to marine pollution.

Animals↗

A serological survey of college students for antibody to Cryptosporidium before and after the introduction of a new water filtration plant.

In April 1997, a large city in the northeastern United States changed their drinking water treatment practices. The city, which previously provided only chlorination for their surface water sources added filtration in addition to chlorination. To assess whether Cryptosporidium infections rates declined following filtration, we tested serological responses to 15/17-kDa and 27-kDa Cryptosporidium antigens among 107 community college students 1 month before and 225 students 5 months after filtration. Results suggest that levels of Cryptosporidium infections did not decline following water filtration. However, seasonal changes in other exposures may have confounded the findings. Swimming in a lake, stream or public pool and drinking untreated water from a lake or stream predicted a more intense response to one or both markers. Residence in the city, not drinking city tap water or drinking bottled water, gender, travel or exposure to pets, young pets, diapers or a household child in day care were not found to be predictive of more or less intense serological responses for either the 15/17-kDa and 27-kDa antigen.

Adolescent↗

Intermolecular electrostatic interactions and their effect on flux and protein deposition during protein filtration.

Although membrane filtration is used extensively to process protein solutions containing a variety of electrolytes, there is currently little fundamental understanding of the effect of the solution environment (and in particular, the solution pH) on the filtrate flux in these systems. We have obtained data for the flux and sieving coefficients during the batch (stirred cell) filtration of solutions of bovine serum albumin, immunoglobulins, hemoglobin, ribonuclease A, and lysozyme through 0.16-micron microfiltration membranes at different pH values. The flux declined significantly for all five proteins due to the formation of a protein deposit on the upper surface of the membrane. The quasi-steady ultrafiltrate fluxes at the individual protein isoelectric pH's were essentially identical, despite the large differences in molecular weight and physicochemical characteristics of these proteins. The flux increased at pH's away from the isoelectric point, with the data well-correlated with the protein surface charge density. These results were explained in terms of a simple physical model in which the protein deposit continues to grow, and thus the flux continues to decline, until the drag force on the proteins associated with the filtrate flow is no longer able to overcome the intermolecular repulsive interactions between the proteins in the bulk solution and those in the protein deposit on the surface of the membrane.

Chemical Phenomena↗

Protein fouling of virus filtration membranes: effects of membrane orientation and operating conditions.

The capacity of virus filters used in the purification of therapeutic proteins is determined by the rate and extent of membrane fouling. Current virus filtration membranes have a complex multilayer structure that can be used with either the skin-side up or with the skin-side facing away from the feed, but there is currently no quantitative understanding of the effects of membrane orientation or operating conditions on the filtration performance. Experiments were performed using Millipore's Viresolve 180 membrane under both constant pressure and constant flux operation with sulfhydryl-modified BSA used as a model protein. The capacity with the skin-side up was greater during operation with constant flux and at low transmembrane pressures, with the flux decline or pressure rise due primarily to osmotic pressure effects. In contrast, data obtained with the skin-side down showed a slower, steady increase in total resistance with the cumulative filtrate volume, with minimal contribution from osmotic pressure. The capacity with the skin-side down was significantly greater than that with the skin-side up, reflecting the different fouling mechanisms in the different membrane orientations. These results provide important insights for the design and operation of virus filtration membranes.

Membranes, Artificial↗

Editorial: The primary glomerular filtration barrier--basement membrane or epithelial slits?

In 1961, based on results obtained with the particulate tracer ferritin, Farquhar, Wissig and Palade [15] proposed a functional model for the glomerulus and defined a role for each of its components in the filtration process: a) the basement membrane as the main filter; b) the endothelium as a valve, which by the number and size of its fenestrae, controls access to the filter; c) the epithelium as a monitor which partially recovers proteins that leak through the filter; and d) the mesangium which serves to recondition and unclog the filter by incorporating and disposing of filtration residues which accumulate against it. In 1966, based on results obtained with the histochemically demonstrable tracers, horseradish peroxidase and myeloperoxidase, Graham and Karnovsky [24] questioned the basement membrane as the site of the main filter and proposed instead that it functioned as a crude prefilter with the epithelial slits representing the final critical barrier. While the concept of the "two-barriers-in-series" has enjoyed wide acceptance, the validity of certain of the experimental data used to support the original hypothesis has been questioned [23, 29, 37]. In the meantime, additional experimental evidence obtained largely with the use of particulate tracers (especially dextrans [23, 43, 44]), has provided strong support for the concept that the basement membrane acts as the main barrier to the passage of molecules in the same size range as plasma proteins (32, 000 to 125,000 mol wt). With respect to the function of the other layers in filtration, additional new information that has come to light has supported the roles proposed above. Work with both particulate [16, 60] and enzymatic [24, 29] tracers, as well as studies by Michael et al [61, 62] with aggregated serum ablumin, supported the phagocytic (unclogging) function of the mesangium. There is evidence from work with particulate tracers (particularly that with dextrans in nephrotic animals [43, 44]) which supports the monitoring function proposed for the epithelium. Recognizing that their work with histochemically demonstrable tracers may have certain technical limitations, Karnovsky, Ainsworth and Schneeberger [29, 37, 38] have recently taken the position that there is no definitive answer to the question of which structure-basement membrane or epithelial slits-represents the principal filter, and have suggested that more information is needed in order to make such a decision. But, in fact, the bulk of the evidence available at present favors the basement membrane as the primary filtration barrier in the glomerulus. Substantial evidence based on work with electronopaque tracers (including recent studies with dextrans) indicates retention of a variety of tracers by the basement membrane. On the other hand, unequivocal demonstration of retention of any tracer by the slits is still lacking.

Animals↗

Gel filtration of sialoglycoproteins.

The role of sialic acid in the gel-filtration behaviour of sialoglycoproteins was investigated by using the separated isoenzymes of purified human liver alpha-L-fucosidase and several other well-known sialic acid-containing glycoproteins (fetuin, alpha1-acid glycoprotein, thyroglobulin and bovine submaxillary mucin). For each glycoprotein studied, gel filtration of its desialylated derivative gave an apparent molecular weights much less than that expected just from removal of sialic acid. For the lower-molecular-weight glycoproteins (fetuin and alpha1-acid glyocprotein), gel filtration of the sialylated molecules led to apparent molecular weights much larger than the known values. The data indicate that gel filtration cannot be used for accurately determining the molecular weights of at least some sialoglycoproteins.

Chromatography, Gel↗

Effects of concentration-polarization on the filtration of proteins through filters constructed from isolated renal basement membrane.

1. Filters comprising multiple layers of rabbit renal tubular basement membrane were constructed with conventional pressure filtration chambers. The effects of concentration-polarization on the behaviour of these filters was assessed by studying the filtration of proteins and of serum under turbulent (stirred) and unstirred conditions. 2. With stirring bovine serum albumin was effectively rejected by the filter barriers (sigma = 0.95) but rejection was diminished (sigma = 0.18) without stirring. The hydraulic permeability of the filters also fell without stirring. 3. In the presence of horse immunoglobulin G, a wholly rejected protein, the rejection of cytochrome c was increased and hydraulic flux was reduced. 4. Filtration studies of serum showed that serum protein was effectively rejected with stirring (sigma greater than 0.999) but rejection diminished when stirring ceased (sigma = 0.98). Albumin was the only protein detected in the filtrate with stirring but alpha- and beta- globulins appeared when stirring ceased. 5. These results show that concentration-polarization markedly affects the behaviour of these basement membrane filters in vitro, since without stirring a polarization layer of rejected protein is formed, which reduces hydraulic permeability and results in increased protein permeation through the filter.

Animals↗