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A solid-phase immunoassay of protease-resistant prion protein with filtration blotting involving sodium dodecyl sulfate.

The precise diagnosis for bovine spongiform encephalopathy (BSE) is crucial for preventing new transmission to humans. Several testing procedures are reported for determining protease-resistant prion protein in various tissues as a major hallmark of prion diseases such as BSE, scrapie, and Creutzfeldt-Jakob disease. However, contamination of materials from tissues or degradation of the specimens sometimes disturbs the accuracy of the assay. Here, we have developed a novel method for solid-phase immunoassay of the disease-specific conformational isoform, PrP(Sc), using filtration blotting of protein in the presence of sodium dodecyl sulfate (SDS) followed by a filtration-based immunoassay with a single anti-prion protein antibody, together with the improved fractionation procedure involving high concentrations of surfactant/detergent. The SDS/heat treatment renders unfolded PrP(Sc) quantitative retention on a polyvinylidene difluoride filter and allows enhancement of the analyte signal with immunodetection; thus, all of the tested specimens are determined with 100% accuracy. In addition, the immunoassay is completed in approximately 1h, indicating its usefulness not only for the screening of BSE specimens but probably also for the postmortem BSE diagnosis of fallen stock as the antibody recognizes the core part of PrP(Sc). The solid-phase immunoassay method, including the filtration blotting with SDS, would be applicable to determining even more sensitively proteins other than PrP(Sc), especially those having rigid conformations.

Animals↗

Clinical effectiveness of leukocyte filtration during cardiopulmonary bypass in patients with chronic obstructive pulmonary disease.

BACKGROUND: We tested the hypothesis that leukocyte filtration during pulmonary reperfusion preserves pulmonary function and results in improved oxygenation after cardiopulmonary bypass (CPB) in patients with chronic obstructive pulmonary disease (COPD). METHODS: In a prospective, randomized study, the treatment group consisted of 20 patients with COPD from consecutive open-heart procedures. A primed leukocyte filter was connected to the arterial line downstream of the standard arterial filter but was excluded from circulation. Circulated blood was directed through the leukocyte filter approximately 10 minutes before aortic cross-clamp removal and at early reperfusion for up to 30 minutes. These patients were compared to 20 additional COPD patients (controls) on whom systemic leukocyte filtration was not used during open-heart surgery. RESULTS: There was no significant difference in gender, age, left ventricular ejection fraction, type of procedure, aortic cross-clamp time, perfusion time, preoperative FEV1 and preoperative respiratory index (Pao2/FiO2 ratio) between treatment and control groups. The respiratory index changed in the treatment group by +9.8% of baseline after completion of CPB, by -14.2% upon arrival in the intensive care unit (ICU), and by -19.6% 12 hours later, whereas in the control group, it changed by -14.5% (p < 0.05), -27.7%, and -24%, respectively. Leukocyte-depleted patients required shorter intubation time (20.4 +/- 16.1 hours), ICU stay (46.2 +/- 40.1 hours) and length of hospitalization (8.3 +/- 2.8 days) than controls (29.5 +/- 21.9 hours, p < 0.05; 75.5 +/- 34.9 hours, p < 0.005; and 10.4 +/- 3.5 days, p < 0.05, respectively). Surgical (30-day) mortality was zero in both groups. CONCLUSIONS: In COPD patients having CPB, systemic leukocyte depletion at early reperfusion was associated with better oxygenation, shorter intubation time, and shorter ICU and hospital stays. Leukocyte filtration during CPB most likely preserves pulmonary function by ameliorating pulmonary reperfusion injury.

Aged↗

Treatment of turkey processing wastewater with sand filtration.

This research investigated the feasibility of coarse/fine sand filtration for removing organic materials from turkey processing wastewater. Sand filtration was tested with three organic and hydraulic loadings. Six two-layer sand bioreactors were in three groups, each with 5 cm layer of pea gravel at the bottom to support layers of fine sand (46 cm) and coarse sand (15 cm) to a height of 66 cm. The bioreactors were inoculated with a mixture of 20% (vol/vol) of wastewater lagoon sludge, 40% (vol/vol) of turkey processing wastewater, and 40% (vol/vol) of BOD(5) dilution water before starting the column operation with turkey processing wastewater. The wastewater contained 1270+/-730 mg COD/L and was applied to each sand bioreactor at hydraulic loading rates of 94% during 80 days of column operation at low and medium hydraulic loading rates (132 L/m(2)/day). The removal at the highest hydraulic loading rate (264 L/m(2)/day) declined after the appearance of a black zone in the top layer of fine sand on day 30 for one reactor and day 50 for the other. The sand filtration in this study represents a feasible treatment for turkey processing wastewater and its efficiency and the life span of the process are associated with the extent of hydraulic loading of the sand bioreactors.

Animals↗

Effects of filtration and pH perturbation on freshwater organic matter fluorescence.

Fluorescence of organic matter from six contrasting freshwaters was analysed after filtration (1.2 microm and 0.2 microm filter sizes) and pH perturbation (+/-2 pH units from ambient conditions). Two fluorophores were compared in detail: tryptophan-like fluorescence, whose filtration and pH characteristics are relatively poorly understood, and humic-like fluorescence, which is better characterised. Although there was some variability in both fluorophores, the tryptophan-like fluorescence showed the most significant decrease in fluorescence intensity between raw and 1.2 microm filter samples, and a much smaller decrease between 1.2 and 0.2 microm, demonstrating a significant source associated with particulate material as well as a significant <0.2 microm fraction. In contrast, humic-like fluorescence shows little change with filtration, suggesting that the majority of this fluorescence is associated with truly dissolved material. The pH perturbation experiments demonstrate that tryptophan-like fluorescence is less impacted by pH than with filter fraction. For humic-like fluorescence, pH effects are weak and are not as consistent as those reported in the literature for extracted humic substances. pH perturbation of the freshwaters shows a wide range of sample specific pH responses, significantly more variable than that observed in experiments using extracted humic substances and tryptophan standards, demonstrating the natural variability of freshwater dissolved organic matter.

Filtration↗

Development of a double-layered ceramic filter for aerosol filtration at high-temperatures: the filter collection efficiency.

The performance of double-layered ceramic filters for aerosol filtration at high temperatures was evaluated in this work. The filtering structure was composed of two layers: a thin granular membrane deposited on a reticulate ceramic support of high porosity. The goal was to minimize the high pressure drop inherent of granular structures, without decreasing their high collection efficiency for small particles. The reticulate support was developed using the technique of ceramic replication of polyurethane foam substrates of 45 and 75 pores per inch (ppi). The filtering membrane was prepared by depositing a thin layer of granular alumina-clay paste on one face of the support. Filters had their permeability and fractional collection efficiency analyzed for filtration of an airborne suspension of phosphatic rock in temperatures ranging from ambient to 700 degrees C. Results revealed that collection efficiency decreased with gas temperature and was enhanced with filtration time. Also, the support layer influenced the collection efficiency: the 75 ppi support was more effective than the 45 ppi. Particle collection efficiency dropped considerably for particles below 2 microm in diameter. The maximum collection occurred for particle diameters of approximately 3 microm, and decreased again for diameters between 4 and 8 microm. Such trend was successfully represented by the proposed correlation, which is based on the classical mechanisms acting on particle collection. Inertial impaction seems to be the predominant collection mechanism, with particle bouncing/re-entrainment acting as detachment mechanisms.

Aerosols↗

Protecting healthcare staff from severe acute respiratory syndrome: filtration capacity of multiple surgical masks.

Guidelines issued by the Centers for Disease Control and Prevention and the World Health Organisation state that healthcare workers should wear N95 masks or higher-level protection during all contact with suspected severe acute respiratory syndrome (SARS). In areas where N95 masks are not available, multiple layers of surgical masks have been tried to prevent transmission of SARS. The in vivo filtration capacity of a single surgical mask is known to be poor. However, the filtration capacity of a combination of masks is unknown. This was a crossover trial of one, two, three and five surgical masks in six volunteers to determine the in vivo filtration efficiency of wearing more than one surgical mask. We used a Portacount to measure the difference in ambient particle counts inside and outside the masks. The best combination of five surgical masks scored a fit factor of 13.7, which is well below the minimum level of 100 required for a half face respirator. Multiple surgical masks filter ambient particles poorly. They should not be used as a substitute for N95 masks unless there is no alternative.

Cross-Over Studies↗

Reduction of ClO2 demand by ClO2 oxidation and subsequent GAC filtration.

The effective removal of soluble natural organic matter (NOM) during water treatment can significantly decrease the ClO(2) demand of processed water. This can be achieved through the oxidation and/or the removal of NOM. The purpose of the study was to examine the influence of ClO(2) oxidation and subsequent GAC filtration on the final ClO(2) demand of treated water. The study showed the ClO(2) demand to be strongly correlated with the abundance of high molecular fractions of NOM in treated water. As it was shown, this part of NOM was effectively removed during the GAC filtration. Moreover, the pre-treatment of water with ClO(2) considerably increased the total capacity of GAC filters for organic and inorganic (i.e. chlorites) oxidation by-products. Therefore, the oxidation of NOM molecules in conjunction with a very efficient GAC filtration can be successfully employed to control the abundance of high molecular NOM components, and thus the ClO(2) demand of finished water.

Chlorine Compounds↗

The use of magnetic bed conditioning and pH control to enhance filtration by natural titanomagnetite.

The fine particle size and magnetic properties of natural titanomagnetite (TM) iron sand offer options for novel granular water filtration systems. The isoelectric points (IEPs) of a variety of natural and synthetic TMs were determined to be in the range from 3.60+/-0.06 to 4.01+/-0.06. TM beds (125-150 microm particle size) expanded by up to 28% were successfully conditioned with vertical fields of approximately 0.018 T and produced up to a three-fold increase in hydraulic conductivity. Filtration studies showed that the filtration efficiency decreased with bed expansion but that this reduction was more than compensated for by reducing the pH to below the IEP of TM. Backwash performance was improved by magnetic conditioning which allowed higher interstitial flow velocities at a given bed expansion.

Electric Conductivity↗

Removal of bulk dissolved organic carbon (DOC) and trace organic compounds by bank filtration and artificial recharge.

Bank filtration and artificial recharge provide an important drinking water source to the city of Berlin. Due to the practice of water recycling through a semi-closed urban water cycle, the introduction of effluent organic matter (EfOM) and persistent trace organic pollutants in the drinking water is of potential concern. In the work reported herein, the research objectives are to study the removal of bulk and trace organics at bank filtration and artificial recharge sites and to assess important factors of influence for the Berlin area. The monthly analytical program is comprised of dissolved organic carbon (DOC), UV absorbance (UVA254), liquid chromatography with organic carbon detection (LC-OCD), differentiated adsorbable organic halogens (AOX) and single organic compound analysis of a few model compounds. More than 1 year of monitoring was conducted on observation wells located along the flowpaths of the infiltrating water at two field sites that have different characteristics regarding redox conditions, travel time, and travel distance. Two transects are highlighted: one associated with a bank filtration site dominated by anoxic/anaerobic conditions with a travel time of up to 4-5 months, and another with an artificial recharge site dominated by aerobic conditions with a travel time of up to 50 days. It was found that redox conditions and travel time significantly influence the DOC degradation kinetics and the efficiency of AOX and trace compound removal.

Berlin↗

Human sperm selection by glass wool filtration and two-layer, discontinuous Percoll gradient centrifugation.

Glass wool filtration and two-layer, discontinuous Percoll (Pharmacia, Uppsala, Sweden) density gradient centrifugation resulted in an average recovery of 50% to 70% of the progressively motile and about 50% of the hypoosmotic swelling (HOS)-positive spermatozoa. Glass wool filtration tended to be more successful than Percoll centrifugation when the ejaculates were asthenozoospermic or produced a suspect/abnormal HOS test. After selection, the acrosin activity increased approximately two- to threefold, but no significant improvement in the percentage of normal sperm forms occurred. Experiments with mixtures of untreated and frozen-thawed ejaculates confirmed that glass wool filtration is more effective in removing nonmotile and HOS-negative spermatozoa than the two-layer Percoll centrifugation technique when the percentage of these types of spermatozoa in the ejaculate is high. The simplicity of these techniques and the good recovery of apparently viable spermatozoa makes these methods more desirable than other, more complicated techniques or procedures that yield a lower recovery of motile spermatozoa.

Cell Separation↗

Assessment of glomerular filtration rate and effective renal plasma flow in cystic fibrosis.

A study was conducted to examine renal function in 10 healthy control subjects and eight patients with cystic fibrosis in stable condition. Sequential bolus injections of 99mTc-DTPA and 125I-OIH were administered to assess glomerular filtration rate and effective renal plasma flow, respectively. Blood was subsequently collected for 3 hours, and urine for 24 hours. Renal clearances of both radioisotope markers were virtually identical in patients and controls. Inasmuch as neither glomerular filtration rate nor effective renal plasma flow was enhanced in patients with cystic fibrosis, increased clearance of drugs in these patients is unlikely to be the result of enhanced glomerular filtration or tubular secretion.

Adolescent↗

Experimental obstructive hydronephrosis in newborn rats. VI. Long-term effects on glomerular filtration and distribution.

Partial obstruction of one ureter was created in newborn rats and its effects were studied after the rats matured. The obstructed pelvis was found to be considerably enlarged. The parenchymal weight was 19 per cent lower and the whole kidney glomerular filtration rate was 43 per cent lower than on the contralateral intact side (p less than 0.001); however, these figures probably include a compensatory increase on the intact side. In the central part of the cortex, the glomerular filtration rate/mg. cortex was 21 per cent lower in the hydronephrotic kidney (p less than 0.02), but there was no intracortical redistribution. The glomerular density in this region was 24 per cent higher on the hydronephrotic side (p less than 0.001), because the glomeruli were crammed together in the distended cortex. No redistribution of glomeruli was found. The results are discussed and compared with previous findings from our laboratory. In this model, partially obstructive uropathy was found to cause only a moderate depression, but no redistribution, of the filtration.

Animals↗

Determination of filtration fraction using the gamma scintillation camera.

A simple technique for determining effective renal plasma flow, filtration fraction and filtration rate is described. The determination depends upon the use of 131iodine labeled hippuran and 99mtechnetium labeled iron ascorbate or Sn diethylenetriaminepentaacetic acid. The filtration fraction is determined by dividing the corrected uptake for the technetium compound by the corrected uptake for radiohippuran. The uptake is simply determined by the 1 to 2-minute count over the kidneys with background subtraction and taking into account the amount injected. The technique is simple, quick and appears to be a useful clinical tool.

Female↗

Diabetes mellitus: polymorphonuclear leukocyte (PMN) filtration parameters and PMN membrane fluidity after chemotactic activation.

The goal of this research was to determine leukocyte rheology at baseline and after chemotactic activation in type I and type II diabetics. In 19 normal subjects, 21 type I diabetics, and 16 type II diabetics at baseline and after in vitro chemotactic activation (prolonged for 5 and 15 minutes) with two stimulating agents (4-phorbol 12-myristate 13-acetate [PMA] and N-formyl-methionyl-leucyl-phenylalanine [fMLP]), we evaluated polymorphonuclear (PMN) filtration parameters (using a St. George filtrometer [Carri-Med, Dorking, UK] and considering the initial relative flow rate [IRFR] and the concentration of clogging particles [CP]) and PMN membrane fluidity (obtained by marking PMNs with the fluorescent probe 1-(4-[trimethylamino]phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). At baseline, there was a difference between normals and type I and II diabetics for PMN membrane fluidity only. After activation in normals and diabetics of both types, a significant variation was present in PMN filtration parameters (IRFR and CP) at both 5 and 15 minutes. In normals, no variation was present in PMN membrane fluidity after activation with PMA or fMLP. After PMN activation, only in type I diabetics was a significant decrease in PMN membrane fluidity present at both 5 and 15 minutes. After PMN activation with either PMA or fMLP in comparison to basal values, only the mean variation (delta%) of the IRFR was significantly different between normals, type I diabetics, and type II diabetics at both 5 and 15 minutes. From the data obtained, it is evident that after activation, the PMN filtration pattern shows a specific behavior in diabetics of both types, while PMN membrane fluidity changes only in type I diabetics. The latter finding may be the basis of a metabolic pattern present in PMNs of this type, revealed after in vitro activation.

Adolescent↗

Treatment of arsenic in Bangladesh well water using a household co-precipitation and filtration system.

Laboratory and field tests were conducted to evaluate the effectiveness of a household filtration process and investigate the effects of phosphate and silicate on the removal of arsenic from Bangladesh groundwater by ferric hydroxides. Fe/As ratios of greater than 40 (mg/mg) were required to reduce arsenic to less than 50 microg/L in Bangladesh well water due to the presence of elevated phosphate and silicate concentrations. The household filtration process included co-precipitation of arsenic by adding a packet (approximately 2 g) of ferric and hypochlorite salts to 20 L of well water and subsequent filtration of the water through a bucket sand filter. A field demonstration study was performed to test the treatment system in seven households in Bangladesh in March and April 2000. Experimental results obtained from the participating families proved that the household treatment process removed arsenic from approximately 300 microg/L in the well water to less than 50 microg/L. The participating families liked this simple and affordable process and used it to prepare clean water for drinking and cooking. A larger scale field test is currently underway.

Adsorption↗

Filtration behaviors of Giardia and Cryptosporidium--ionic strength and pH effects.

The laboratory-scale filtration tests of Giardia cysts and Cryptosporidium oocysts in both 2 mm-phi glass beads and 2 mm-phi polystyrene beads filters were conducted to investigate their filtration behaviors. The protozoan parasites were used as target particles, while the chemical system altered by changing the electrolyte concentration and pH. The results significantly indicate that ionic strength have a positive effect on the removal efficiencies for Giardia cysts and Cryptosporidium oocysts. The removal efficiency of two filters for Giardia cysts slightly decreased from pH 2.4 to 8.7 and decreased significantly in pH as pH up to 8.7, while that for Cryptosporidium slightly rippled beyond pH 8.7, and with the decrease in pH up to pH 8.7. The experimental collision efficiencies from the interactions between colloids and the filter media were calculated with a semi-empirical approach of the single sphere model and clean-bed filtration theory. The results also indicated that experimental collision efficiencies for (oo)cysts corresponded to the (oo)cysts removal efficiencies in all trials, and oocysts exhibits higher collision efficiencies than cysts.

Animals↗

Factors affecting seasonal variation of membrane filtration resistance caused by Chlorella algae.

A seasonal fluctuation pattern was observed in membrane filtration resistance by Chlorella algae cultured in open ponds in the tropical environment. In order to investigate the causes of this phenomenon, Chlorella was cultivated under controlled conditions and the cake resistance was measured by batch filtration in dead-end mode. The filtration resistance was found to be a function of environmental conditions. Algae could grow favourably and offered low specific cake resistance (R,s) on the order of 10(11) m/g for the culture temperature from 28 degrees C to 35 degrees C. The algal growth was inhibited and the specific cake resistance increased to the order of 10(12) m/g below or above this optimum temperature range. Strong solar radiation, coupled with high temperatures, also inhibited the growth of algae and resulted in higher specific cake resistance. The specific cake resistance of algae cultured at different temperatures increased with the amount of the extracellular organic matter (EOM) extracted by 0.1 N NaOH. Hence EOM, rather than bacteria present in the mono-algal culture, was considered to be the primary factor affecting the cake resistance. The specific cake resistance increased drastically after actively growing cells were stored in nutrient-free water under dark conditions. However, the resistance was slightly decreased when the algal cells were stored in NSIII nutrient media in a dark room, indicating the effect of nutrient availability on the change of the specific cake resistance under the light-limiting conditions. EOM extracted from the cells kept in the nutrient-free water contained less sugar than the fresh culture, whereas the EOM extracted from the cells stored in the NSIII media contained more sugar. The molecular distribution of the EOM shifted from below 1,000 kDa before storage to more than 2,000 kDa after storage in both the nutrient-free and NSIII media.

Chlorella↗

Advanced treatment of sewage by pre-coagulation and biological filtration process.

A pre-coagulation and bio-filtration process for advanced treatment of sewage was developed and experimentally discussed with a pilot plant. The bio-filtration unit consists of a denitrification filter, a nitrification filter with side stream to the denitrification filter, and a polishing filter with anoxic and aerobic parts. Concentrations of SS, T-COD(Cr), T-carbonaceous BOD, T-N and T-P in the effluent were stably kept at less than 3, 20, 5mg/L, 2mg N/L and 0.2mg P/L, respectively, and transparency at higher than 100 cm, under total hydraulic retention time of 3.2h in the bio-filtration parts (filter-bed). ORP in an anoxic tank before a nitrification tank should be at a low level of less than -120 mV to keep remaining NO(-)(x) - N less than 1mg N/L, but must be maintained at a level higher than -150 mV. The maximum nitrogen-loading rate under a water temperature of 18 degrees C should be less than 0.25 kg N/(m(3)-filter-bed.d). Concentrations of microorganisms kept in the reactors were as high as 4000-5000 mg COD/L-filter-bed. Denitrification activity of 0.4 or 0.7 kg N/(m(3)-filter-bed.d), and nitrification activity of 0.3 kg N/(m(3)-filter-bed.d) were obtained, respectively, under a water temperature of about 18 degrees C. Backwashing in each tank as well as methanol addition and aeration in the polishing filter were operated successfully by the automatic control systems. These results proved that this process is applicable to advanced treatment of sewage with easy maintenance.

Automation↗