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Comparison of membrane filtration methods for the recovery of legionellae from naturally contaminated domestic drinking water supplies.

We compared four methods for the cultural recovery of legionellae from naturally contaminated water samples: (A) inoculating 1 and 0.1 ml directly on the selective solid growth medium, (B) filtration through cellulose nitrate membrane filters and inoculating the resuspended residue corresponding to 100 and 10 ml of the original sample on the medium, (C) filtration through polycarbonate filters and inoculating the resuspended residue corresponding to 100 and 10 ml of the original sample, and (D) filtration of 100 and 10 ml through cellulose nitrate filters and incubating the filter directly on the medium. Of the water samples tested in parallel, 103 samples tested positive for legionellae in at least two of these methods. The mean number of CFU/ml recovered by methods A, B, C, and D werte 5.45, 1.85, 1.70, and 2.26, respectively. The differences between the methods proved to be highly significant except for methods B and C (p = 0.112), the results of which correlated closely with each other (r = +0.96). In comparison with method A the recovery rates of methods B, C, and D were found to be 60, 57, and 69%, respectively. We recommend the use of method A for the recovery of legionellae at concentrations of > or = 1 CFU/ml. If legionellae below this concentration are to be detected we recommend method D as the easiest and most sensitive technique.

Collodion↗

Microvascular fluid filtration capacity (Kf) assessed with cumulative small venous pressure steps and with various degrees of tilt.

UNLABELLED: Orthostatic dysregulation is a frequent phenomenon in pilots experiencing extreme G forces and after prolonged exposures to microgravity in cosmonauts. We used non-invasive venous congestion plethysmography (VCP) to study microcirculatory changes and the ability to activate peripheral and centrally mediated protective mechanisms during various degrees of tilt, which we used as an orthostatic challenge. METHOD: The study, which was approved by the local ethical committee of the Institute of Biomedical Problems, was performed on six healthy 20 - 26 years old male volunteers. We applied 6 - 8 cumulative small venous congestion pressure steps (8 mmHg) to the thigh and determined the fluid filtration capacity (Kf), the linear relationship between cuff pressure (Pcuff) and measured fluid filtration (Jv). We then measured the fluid filtration (Jv) response to varying cumulative degrees of tilt, starting at 0 degrees followed by head down -8 degrees -15 degrees, -30 degrees -15 degrees, -8 degrees, 0 degrees, and then head up 15 degrees, 30 degrees, 70 degrees, 30 degrees, 15 degrees and 0 degrees. Each tilt stage was sustained for 15 minutes. The change in hydrostatic load, at the level of the strain gauge, was determined by measuring the difference in vertical height between the right atrium and mid calf at the level of the gauge. Limb arterial blood flow was measured and lung function tests were performed before and after the small cumulative pressure step protocol, as well as at the end of each tilt step. RESULTS: No significant changes in blood pressure (BP) and heart rate (HR) occurred during the cumulative pressure step protocol. However, an increase in HR was observed during the tilt, but only reached significance at 30 degrees and 70 degrees. The mean Kf value measured with small cumulative pressure steps was 3.25 +/- 0.5 (10(-3) ml.100 ml tissue(-1) mmHg(-1) = KfU), which was significantly (p < 0.005) higher than the value obtained using tilt to increase the hydrostatic load (0.98 +/- 0.2 KfU). One subject had an unchanged Kf value and experienced vaso - vagal syncope following the imposition of 70 degrees tilt, with a heart rate < 35 / min and a systolic blood pressure of 60 mmHg. The measurement of blood flow in the limb showed no significant change during the tilt procedure. The lung function measurements revealed, that only expiratory reserve volume (ERV) did significantly increase during 35 degrees and 70 degrees head up tilt. All other parameters were unchanged. We suggest that the differences in these Kf values can be explained by the activation of both central and peripheral mediated changes in pre- capillary resistance in response to the tilt. We suspect, that the vaso - vagal syncope, experienced by one subject, reflected his inability to activate these protective reflex mechanisms, a situation that could be exacerbated by sub-clinical venous insufficiency. We believe that these results show that VCP is a useful tool for the determination of intolerance to orthostatic stress.

Adult↗

[The effect of heterogeneous scleral transplantation on hypotony after filtration surgery of glaucoma with mitomycin C].

OBJECTIVE: The study was designed to observe the effect of heterogeneous scleral transplantation in treatment of hypotony after filtration surgery of glaucoma with mitomycin C. METHOD: 7 patients (8 eyes) with hypotony after filtration surgery of glaucoma with mitomycin C were treated by heterogeneous scleral transplantation. RESULTS: The post-operative follow-up periods ranged from 3 to 19 months (mean 7.75 +/- 5.37 months). Postoperatively, the visual acuity was increased in 5 eyes, and the intraocular pressure was increased to normal level in all eight eyes. CONCLUSION: Heterogeneous scleral transplantation has therapeutic effects in the treatment of hypotonic eyes after filtration surgery of glaucoma.

Adolescent↗

Membrane filtration for virus removal.

Incorporation of adequate clearance methodologies for viral safety assurance constitutes an integral part of the manufacturing process for plasma-derived biologicals. Filtration, being non-invasive, non-destructive and a < > mechanism for virus removal is often the method of choice. This paper discusses the virus filter performance characteristics and other process-related issues specifically applying to the Pall Ultipor VF filters (Grade DV50 and Grade DV20), which are operated in the direct flow filtration (DFF) mode and the Pall Filtron Omega VR filters (the Omega 300 K and Omega 100 K), which are operated in the tangential flow filtration (TFF) mode.

Animals↗

[Application of mitomycin C in glaucoma filtration surgery].

OBJECTIVE: To investigate the efficacy and the complication of glaucoma filtration surgery with intraoperative mitomycin C. METHODS: Between April 1994 and February 1996, 33 eyes of 28 patients underwent filtration procedures with intraoperative mitomycin C. Then a follow up of mean 6.65 months (range, 1-12 months) was taken. RESULTS: The Class I success rate was 72.7% when the intraocular pressure (IOP) was less or equal to 2.79 kPa without glaucoma medicine. The Class II success rate was 24.2% when the IOP was controlled with topical glaucoma medicine. In the postoperative period, persistant hypotony developed in 4 eyes, and hypotony-related maculopathy developed in 2 of these eyes. No conjunctival wound leak and endophthalmitis was present. CONCLUSIONS: Intraoperative mitomycin C in glaucoma filtration surgery can increase the success rate, while it is associated with a higher incidence of complications such as hypotony.

Adolescent↗

[Influence on tear film of postoperative 5-fluourouracil and intraoperative mitomycin C in glaucoma filtration surgery].

OBJECTIVE: To investigate the changes of tear film functions due to the use of postoperative 5-fluourouracil (5-Fu) and intraoperative mitomycin C (MMC) in glaucoma filtration surgery. METHODS: 71 eyes of 71 refractory glaucoma patients were followed up after filtration surgery. They were divided into 3 groups. The 5-Fu and MMC groups consisted of 24 and 17 eyes respectively. The other 30 eyes that underwent the surgery without any use of antimetabolic agents were in the control group. The observed symptoms included ocular foreign body sensation, dryness, photophobia, itching and discharge. The available criteria were conjunctival congestion, fluorescein and rose bengal staining on the cornea and conjunctiva, break-up time of tear film and Schirmer's test. RESULTS: The ocular symptoms and signs were manifest and tear film functions were significantly decreased in 5-Fu group (P < 0.05). No significant difference between MMC and the control was seen, except that the dry feeling was significantly intensive in MMC group (P < 0.05). CONCLUSION: The postoperative use of 5-Fu after glaucoma filtration surgery leads to tear film abnormalities apparently.

Adult↗

Functional abnormalities of human neutrophils collected by continuous flow filtration leukopheresis.

Continuous flow filtration leukopheresis (FL) is a relatively simple, inexpensive, and efficient technique of harvesting blood neutrophils from normal donors for transfusion into neutropenic recipients. There has been concern, however, that neutrophils may be functionally altered druing this leukopheresis procedure. Human neutrophils obtained by various FL techniques were studied for in vitro chemotaxis by a 51Cr-radiolabel method and for in vitro killing and phagocytosis of Staphylococcus aureus. We compared their function with neutrophils obtained by the NCI-IBM cell separator and by dextran sedimentation from whole blood. FL neutrophils eluted from nylon filters after 3-hr collection periods were functionally abnormal by all parameters tested, while neutrophils obtained by cell separator after similar collection times were not significantly different from control cells. However, neutrophils from 3-hr FL collections were found to include both normal and abnormal populations of cells. Loosely adherent cells, eluted after tapping the filters, were functionally normal; more adherent cells, eluted after tapping the filters and representing the bulk of cells collected, were progressively more abnormal the less readily they were eluted. Shortened FL collection times (1-2 hr) were found to decrease the functional defects. Also, administration of dexamethasone to donors prior to filtration leukopheresis diminished the functional defects of FL neutrophils perhaps by altering adherence characteristics of the cells. These studies show that neutrophils obtained by filtration leukopheresis are functionally abnormal in relation to the time and extent of adherence to nylon filters.

Adult↗

[The renaturation and purification of RGD-staphylokinase by gel filtration].

A recombinant RGD-Staphylokinase(RGD-Sak) with thrombolytic and anti-thrombolytic bifunction was expressed in E. coli. The expression product accumulates as inclusion bodies. In order to obtain active molecule, the RGD-Sak in the inclusion body should be denatured and then renatured. The renaturation of RGD-Sak was performed by gel filtration. Comparing with the traditional way of dilution renaturation, gel filtration way is better than the traditional one, since there are some advantages, such as simple processing, high recovery, low cost and higher purity after renaturation, After renaturation, RGD-Sak was purified by Q-Sepharose FF, and the purity was more than 95%. Analysis of CD spectra showed that the final product from the two renaturation ways have similar CD spectra. It was demonstrated that RGD-Sak molecules proceeded correct refolding through gel filtration or dilution renaturation process.

Chromatography, Gel↗

[Application of dot-immunogold filtration assay in detection of IgG in sera of trichinellosis patients].

OBJECTIVE: To study a new quick method for detecting serum IgG against Trichinella spiralis (T.s). METHODS: Membrane antigen of muscle larva of T.s was isolated and combined with the nitrocellulose membrane. Goat anti-human IgG was conjugated with the golden pellets of colloidal state which was used as marked antibody for the experiment of dot-immunogold filtration assay to examine specific IgG. RESULTS: T.s IgG and its corresponding antigen reacted on the membrane by filtration, and the result could be observed with naked eyes within 10 min. In the examination of 76 clinical serum samples, the positive rate was 94.7%. It was not significantly higher than that of ELISA (chi 2 = 2.83, P > 0.05). CONCLUSION: The dot-immunogold filtration assay is rapid and simple for performing and reading in the detection of IgG against T.s with high sensitivity and specificity.

Animals↗

Decolorization of heat-treatment liquor of waste sludge by a bioreactor using polyurethane foam-immobilized white rot fungus equipped with an ultramembrane filtration unit.

A bench-scale bioreactor using immobilized fungal cells equipped with an ultramembrane filtration unit was developed as a means of decolorizing brown color components (melanoidins) arising from the heat-treatment liquor (HTL) of waste sludge. Artificial HTL containing 4200 color units of synthetic melanoidin supplemented with 1000 mg/l ethanol was first subjected to decolorization by the fungus Coriolus hirsutus IFO4917 immobilized onto polyurethane foam cubes. Then, the resultant biologically treated HTL was subjected to ultrafiltration to obtain the permeate (filtrate) as the effluent. The retentate (concentrate) of the filtration unit, containing the remaining melanoidin of high molecular weight and extracellular decolorizing enzymes, was returned to the fungal bioreactor to allow further decolorization. This system was operated in a sequencing batch mode under nonsterile conditions. Contamination of the bioreactor with air/water-born microbes markedly lowered the decolorization efficiency. However, this problem was solved by heating the returned concentrate at 50 degrees C for 10 min. Under the almost stable condition of a hydraulic retention time of 2 d in a 1-d cycle sequencing batch mode, about 70% decolorization was routinely achieved using the entire system (bioreactor + ultrafiltration), while the contribution of the fungal bioreactor alone to the decolorization was about 45%.

Journal Article↗

[A case of severe glaucoma with pseudopemphigoid successfully treated by filtration surgery using amniotic membrane].

BACKGROUND: It is necessary to decrease topical anti-glaucoma medication for severe glaucoma with pseudopemphigoid caused by anti-glaucoma eye drops. Glaucoma filtrating surgery is often needed instead of medication, but the prognosis is poor because it induces scar fomation and makes the filtrating bleb vanish. CASE: An 85-year-old male patient with exfoliation syndrome had twice undergone glaucoma surgery about ten years previously. His intra-ocular pressure (IOP) was high despite topical anti-glaucoma medication. At the first examination in our hospital, he had severe superficial punctate keratopathy, blephariticshortening and symblepharon, and we therefore diagnosed severe pseudopemphigoid induced by anti-glaucoma eye drops. Because his IOP could not be controlled by topical and general medication, we conducted a glaucoma filtrating operation using amniotic membrane. CONCLUSION: The administration of oral anti-inflammatory drugs before and after surgery and the use of amniotic membrane prevented post-operative scar formation and the progress of symblepharon, resulting in the successful control of IOP after surgery.

Aged, 80 and over↗

Antioxidant and anti-inflammatory activity of extract obtained from Aspergillus candidus MTCC 2202 broth filtrate.

Antioxidant potential of Aspergillus candidus MTCC 2202 broth filtrate extract was studied using different antioxidant models, whereas anti-inflammatory potential was studied using carrageenan-induced rat paw oedema model. The ethyl acetate extract at 1000 microg/ml showed maximum scavenging activity of the stable radical 1,1-diphenyl,2-picryl hydrazyl upto 96.65% (IC50=430.36 microg/ml) and scavenging of the radical cation, 2,2-azinobis-(3-ethylbenzothiazoline-6-sulphonate) upto 92.25% (IC50=606.29 microg/ml) at the same concentration. The extract had good reducing power, however showed moderate inhibition for conjugated dienes and thiobarbituric acid reactive acid substances (59.56 and 51.45%). The total phenolic content of various extracts of A. candidus broth filtrate was measured and a correlation between radical scavenging activities of extracts with total phenolic content was observed. The ethyl acetate extract (125 mg/kg ip) showed significant anti-inflammatory activity in carrageenan-induced rat paw oedema model. The exhibited antioxidant activity of ethyl acetate extract of A. candidus broth filtrate was comparable with BHA and ascorbic acid, while anti-inflammatory activity was comparable with standard diclofenac sodium.

Animals↗

Measurement of capillary filtration coefficient in a solid tumor.

The net transvascular filtration rate, JF (ml/min/100 g), in an isolated, RBC-free perfused R3230AC mammary adenocarcinoma tumor was measured using a gravimetric method whereby changes in tissue weight over time were monitored. From the gravimetric measurements of JF following changes in venous pressure, the capillary filtration coefficient (ml/min/mm Hg/100 g) was found to be 2.2 (range, 0.8-2.8), i.e., 10 to 1000 times higher than those found in several normal tissues and within the range of those reported for glomerular capillaries. These measurements of capillary filtration coefficient are consistent with elevated tumor interstitial fluid pressure, interstitial fluid flow, and peritumor edema.

Adenocarcinoma↗

[Histologic comparison of a functioning and non-functioning filtration membrane with the Molteno implant].

In two patients with a Molteno implant, the filtration membrane was excised 2 and 21 months postoperatively because of uncontrolled elevation of intraocular pressure. This tissue was examined by light and electron microscopy and the findings compared with those of a third functioning Molteno filtration membrane obtained from an enucleation specimen. The most conspicuous differences were the high density and cellularity of the connective tissue of the non-functioning membranes and the frequent cell pyknosis and necrosis in the functioning membrane. In addition, one of the non-functioning membranes showed a condensed cellular layer on the inner surface. There were no inflammatory cell infiltrates in any of the three specimens. We conclude that the variable results of Molteno implants are related to the quantities of tissue components of the filtration membrane.

Adult↗

[Clinical hemorheologic study of 65 inflammatory syndromes explored using standard biologic examination and total blood filtration test].

The hemorrheologic impact of an inflammatory syndrome was investigated in a prospective biological study of 65 patients by using an erythrocyte filtration test on total blood. Impact was assessed by a statistical study of correlations between relative filtration time (RFT), inflammation-reactive proteins (IRP) and the different parameters of the hemogram. A stepwise multiple regression test (SMRT) indicated the dominant role of fibrinogen in determining acceleration of the sedimentation rate (p less than 10(-3) and lengthening of RFT (p less than 10(-3). Moreover, RFT appeared to be very significantly correlated with polymorphonuclears (R = 0.55; p less than 10(-3] hematocrit as measured by the centrifugation technique (R = 0.34; p = 0.005) and corpuscular concentration in hemoglobin (CCHM) (R = 0.30; p = 0.01). A chart predictive of microcirculatory risk was deduced from the study based on fibrinogen values and hematocrit. However, the relatively low rate of reduction in the total variance observed during SMRT suggests the quite relative role of the studied parameters in erythrocyte filtration disturbances during an inflammatory syndrome.

Blood Sedimentation↗

[Double filtration plasmapheresis in the treatment of vascular complications of hyperlipidemia].

Double-filtration plasmapheresis is a therapeutic procedure for the extracorporeal depuration of atherogenic lipoproteins, which does not require the administration to the patient of exogenous fluids. We have used it in two patients affected by hyperlipidemia with severe cardiovascular complications. Both patients presented a dramatic improvement of their symptoms (angina pectoris and claudicatio intermittens) shortly after the beginning of treatment. By the brisk reduction of circulating low-density lipoproteins, plasma-filtration may favor the removal of cholesterol from atheromatous plaques of vessel walls. Furthermore, this procedure may modify platelet aggregation and blood viscosity. Our observation suggests that plasma-filtration may be useful not only for delaying coronary heart disease in the rare cases of homozygous familial hypercholesterolemia, but also in the management of patients with other primary hyperlipoproteinemias and clinical manifestations of already established cardiovascular complications.

Adult↗

Glomerular filtration and tubular secretion of MAG-3 in the rat kidney.

Technetium-99m mercaptoacetyltriglycine (MAG-3) has recently been introduced as a new radiopharmaceutical for dynamic renal scintigraphy. To elucidate the mechanism of renal excretion, micropuncture experiments were performed in rat kidneys for direct measurements of glomerular filtration and tubular secretory capacity. Fluid of Bowman space was collected from superficial glomeruli and analyzed for its contents of [99mTc]MAG-3, [125I]hippurate and [3H]inulin during constant infusion of these compounds. The ratio of activity of ultrafiltrate to that of arterial plasma was 0.23 for MAG-3, 0.68 for hippurate and 1.04 for inulin which demonstrates that the filtrated amount of MAG-3 is only 23% of that of inulin, presumably because of higher plasma protein binding which was also measured in vitro and found to be 80 +/- 1.5% for MAG-3 and 32 +/- 2% for [125I]hippurate. Proximal and distal tubules were also micropunctured and their tubular fluid as well as the final urine analyzed for the activity of hippurate and MAG-3. The tubular fluid to plasma ratio values along the nephron and in the final urine were all lower for MAG-3 than for hippurate, indicating a lower secretory capacity. From measurements of whole renal clearance, GFR and plasma protein binding the filtered amount of MAG-3 was 0.26 and of hippurate 0.87 ml/min.g kidney weight (p less than 0.001) and the secreted amount 2.01 and 2.38 ml/min.g kidney weight (p less than 0.05), respectively. We conclude that MAG-3 is predominantly excreted by tubular secretion and that the lower renal clearance of MAG-3 as compared with that of hippurate is a result both of a substantially decreased glomerular filtration and of a lower tubular secretion.

Animals↗