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A new approach for the filtrate regeneration system in the wearable artificial kidney.

In the filtrate and/or dialysate regeneration system, which is expected to miniaturize the artificial kidney, ammonium ion decomposed from urea by immobilized urease is removed competitively by ion-exchangers from coexisting cations. Since divalent cations such as Ca2+ and Mg2+ are more favorably exchanged than ammonium ion, this system needs supplementation of these cations and, thus, additional amount of ion-exchanger. To minimize these requirements, we utilized positively charged membrane to process cation-free filtrate in which urea is dialyzed. Positively charged membranes were tested in vitro to evaluate separation efficiency between urea and cations. Equilibrium adsorption of ammonium ion to ion-exchanger with or without co-existing cations demonstrates that this filtrate regeneration system can reduce the amount of ion-exchanger up to one-half to one-third of that of the conventional system. In an ex vivo experiment with a mongrel dog, blood urea nitrogen (BUN) concentration was maintained at the same level or up to 23% less than the initial value.

Aluminum Silicates↗

Comparative evaluation of iohexol and inulin clearance for glomerular filtration rate determinations.

We evaluated iohexol as a filtration marker in 150 children. The clearance of iohexol was compared with that of inulin or with a formula clearance. The single-sample clearance of iohexol showed a good correlation with the clearance of inulin (r = 0.834). The clearance of iohexol correlated well (r = 0.672) with the formula clearance. The optimal blood sampling time for iohexol clearance determinations appears to be between 120 and 180 min after injection, at least in patients with relatively normal filtration rates. We conclude that iohexol clearance is an accurate method of determining the glomerular filtration rate in clinical practice.

Child, Preschool↗

Intrarenal distribution of blood flow and glomerular filtration during chronic unilateral ureteral obstruction.

Paired hydronephrotic (HN) and hypertrophic (HT) rat kidneys were studied after 6 days with complete unilateral ureteral obstruction without exposing the kidneys. Total HN renal blood flow (RBF), estimated by total microsphere (MS) uptake and from local 125I-antipyrine (Ap) uptake, averaged about 3/4 of control. HN kidney GFR was reduced to about 1/2 of control level as estimated from inulin clearance of HT kidney times the HN to HT ratio for mean single nephron filtration rate, determined by 14C-ferrocyanide. Whereas blood flow (Ap) was proportionately reduced in outer and inner cortex (OC and IC), fractional flow to the outer medulla (OM) was doubled as compared to controls (p less than 0.01). Filtration was well preserved in deep as compared to superficial glomeruli with a smaller deep nonfiltering fraction (p less than 0.02). Thus the results oppose the current concept that HN is characterized by disproportionate circulatory damage to IC and OM with little or no filtration in deep nephrons. In HT kidneys average RBF (MS) and GFR rose by about 1/2. Whereas total blood flow (Ap) rose proportionately in OC and IC, it remained at control level in OM, indicating dissociation between the total RBF and GFR and the effective blood flow to the OM zone.

Animals↗

Total and local renal blood flow and filtration in the rat during reduced renal arterial blood pressure.

Discrepancies concerning the lower pressure limit of renal blood flow (RBF) autoregulation, different autoregulatory adjustments in deep and superficial renal zones and dissociation of RBF and filtrate production stimulated the present study. Autoregulation of renal blood flow was investigated at reduced renal arterial blood pressure (RAP) in Sprague-Dawley rats using 4 different flow methods: uptake of 125I-iodoantipyrine (I-Ap) and 86Rb, local detection of hydrogen gas washout rate (H2) and, in the autoperfused kidney, electromagnetic flowmetry (FM). With I-Ap and 86Rb, RBF was maintained at a RAP as low as 80 mmHg, compared to contralateral RBF. However, with the other two methods where each kidney serves as its own control, a 15% RBF reduction was obtained at this RAP. This discrepancy (p less than 0.001) infers a contralateral renal vasoconstriction during ipsilateral renal hypotension and vasodilation. Arterial blood pressure increased during unilateral renal hypotension, suggesting that contralateral renal constriction was part of a general increase in total body vascular resistance. Following abrupt RAP reduction RBF was immediately readjusted (2-3 s) and maintained for up to 40 min. No significant change in intrarenal blood flow distribution was observed with I-Ap. Superficial and deep cortical single nephron glomerular filtration rates were equally reduced at lowered RAP as determined by the ferrocyanide technique. However, a dissociation between the autoregulation of RBF and glomerular filtration rate (GFR) in the direction of less well maintained GFR was observed. Renal arterial acetylcholine infusion increased RBF by about 40% and effectively abolished RBF autoregulation.

Acetylcholine↗

Time-dependent heterogeneity of filtration rate in the autoregulating rat kidney.

Experiments were performed to study the regulation of the single-nephron glomerular filtration rate (SNGFR) in superficial and juxtamedullary nephrons, as the left kidney of Sprague Dawley rats was submitted to a reduced arterial pressure of 70 mmHg by means of an aortic clamp. The SNGFR at different cortical levels was measured 0.5, 1, 5, 20 or 45 min after the reduction, in order to ascertain whether the effects of the regulatory mechanisms are modified with time. A Hanssen technique was used, which allows one determination of filtration rates per animal. At a renal arterial pressure (RAP) of 100 mmHg (= control animals) the SNGFR amounted to 20 +/- 1.2 and 23 +/- 0.8 nl X min-1 X g-1 kidney weight in the outer and inner cortical (OC, IC) nephrons. When RAP was further reduced to 70 mmHg, the autoregulation of SNGFR, determined after 0.5 min, was highly efficient for both OC and IC nephrons (19 +/- 2.0, 23 +/- 2.6). A prolonged reduction in RAP caused a gradual decline in SNGFR. The filtration rate measured after 5 min was 15 +/- 1.4 for OC and 20 +/- 1.8 for IC nephrons. The decline was most pronounced for OC nephrons, which led to a fractional redistribution in favour of IC nephrons. Thus, SNGFRIC/SNGFROC was 1.16 +/- 0.065 when RAP was 100 mmHg and 1.41 +/- 0.126 after 5 min with an RAP of 70 mmHg. It is well documented that suprarenal aortic occlusion is a powerful stimulus for the release of renin. This was manifested as an increase in the arterial pressure proximal to the aortic clamp.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Angiotensin II and renal prostaglandin release in the dog. Interactions in controlling renal blood flow and glomerular filtration rate.

The relationship between angiotensin II and renal prostaglandins, and their interactions in controlling renal blood flow (RBF) and glomerular filtration rate (GFR) were investigated in 18 anaesthetized dogs with acutely denervated kidneys. Intrarenal angiotensin II infusion increased renal PGE2 release (veno-arterial concentration difference times renal plasma flow) from 1.7 +/- 0.9 to 9.1 +/- 0.4 and 6-keto-PGF1 alpha release from 0.1 +/- 0.1 to 5.3 +/- 2.1 pmol min-1. An angiotensin II induced reduction in RBF of 20% did not measurably change GFR whereas a 30% reduction reduced GFR by 18 +/- 8%. Blockade of prostaglandin synthesis approximately doubled the vasoconstrictory action of angiotensin II, and all reductions in RBF were accompanied by parallel reductions in GFR. When prostaglandin release was stimulated by infusion of arachidonic acid (46.8 +/- 13.3 and 15.9 +/- 5.4 pmol min-1 for PGE2, and 6-keto-PGF1 alpha, respectively), angiotensin II did not change prostaglandin release, but had similar effects on the relationship between RBF and GFR as during control. In an ureteral occlusion model with stopped glomerular filtration measurements of ureteral pressure and intrarenal venous pressure permitted calculations of afferent and efferent vascular resistances. Until RBF was reduced by 25-30% angiotensin II increased both afferent and efferent resistances almost equally, keeping the ureteral pressure constant. At greater reductions in RBF, afferent resistance increased more than the efferent leading to reductions in ureteral pressure. This pattern was not changed by blockade of prostaglandin synthesis indicating no influence of prostaglandins on the distribution of afferent and efferent vascular resistances during angiotensin II infusion. In this ureteral occlusion model glomerular effects of angiotensin II will not be detected, and it might well be that the shift from an effect predominantly on RBF to a combined effect on both RBF and GFR induced by inhibition of prostaglandin synthesis is located to the glomerulus. We therefore postulate that renal prostaglandins attenuate the effects of angiotensin II on glomerular surface area and the filtration barrier, and not on the afferent arterioles as previously suggested.

Angiotensin II↗

Filtration blebs in corneoscleral wounds sutured with Dexon 7-0 and Dexon 8-0.

In a retrospective investigation the incidence of post-operative subconjunctival filtration with bleb formation were studied in 112 eyes which had undergone cataract surgery with a corneoscleral incision sutured with braided polyglycolic acid sutures (Dexon). In 61 consecutive cataract operations sutured with Dexon 8-0 there were filtration blebs in 24 (39,3%) eyes, and in 51 consecutive cataract operations sutured with Dexon 7-0 there were filtration blebs in 11 (21,6%), eyes. Although there is a difference between the two materials, this is not statistically significant with the normal security level P less than 0.05.

Aged↗

Concealed glomerular filtration.

1. An increase in apparent renal clearances is frequently observed on restoring urine flow after a period of anuria or on increasing it after oliguria. An analysis of such 'peaks' in clearance has been made in experiments on anaesthetized dogs, using two preparations of labelled vitamin B12 and urine collections of 1-2 min. [57Co]B12 was infused throughout the experiments, while [58Co]B12 was given as a single injection during periods of anuria or oliguria induced by noradrenaline infusion, haemorrhage or aortic obstruction. 2. The apparent high clearance in the first minute or two of restored or increased flow is an artifact explained by inclusion in the peak of material filtered earlier, but not excreted. By means of the integrated plasma concentration ratio of the two B12 isotopes during the period of low or absent flow, the excess B12 in the peak may be reapportioned between the period before the 58Co was injected and the period after it. 3. The findings indicate that filtration may temporarily continue during anuria, but this is concealed as a result of failure of onward flow of filtrate. In oliguria a similar concealment of filtration may result from the cessation of onward flow in some nephrons.

Animals↗

Factors affecting the maturation of glomerular filtration rate and renal plasma flow in the new-born dog.

1. The maturation of glomerular filtration rate was studied by comparison of thirty-six new-born mongrel dogs aged 1-35 days with six adult dogs.2. Under mannitol diuresis, glomerular filtration rate (GFR) rose from 0.16 ml. min(-1).g kidney(-1) at 1 day of age to 0.34 ml. min(-1).g kidney(-1) at 1 month of age. Adult GFR averaged 0.68 ml. min(-1).g(-1). There was good correlation of GFR with arterial blood pressure (r = 0.76, P < 0.001). Part of the statistical correlation of GFR with blood pressure was found to be independent of the relationship between blood pressure and age.3. Acute increases or decreases in blood pressure resulted in parallel changes in GFR in the puppies. There was no change of GFR with change of blood pressure in adult dogs. Carotid artery clamping, independent of blood pressure changes, produced increased renal vascular resistance and decreased GFR in the pups.4. Renal plasma flow (RPF) increased from 0.70 ml. min(-1).g(-1) at 1 day of age to 1.80 ml. min(-1).g(-1) at 1 month and showed good correlation with blood pressure (r = 0.67, P < 0.001). Filtration fraction (GFR/RPF) and renal vascular resistance did not vary with age in the pup and were the same as those for the adult.5. These results support the hypothesis that maturation of GFR and RPF are closely related to maturation of arterial blood pressure in the mongrel dog. The factors other than blood pressure which also affect renal maturation in the dog still need to be more clearly defined.

Age Factors↗

Microvascular pressures and filtration coefficients in the cat mesentery.

1. Filtration coefficient and hydrostatic pressure have been measured in single capillaries and venules in the cat mesentery using a modification of the Landis (1927) single vessel occlusion technique. 2. Venules were found to be filtering fluid, not absorbing it as is often supposed. 3. The mean filtration coefficient in capillaries was 0.018 micrometers . s-1 . mmHg-1 (1.35 X 10(-10)m . s-1 . Pa-1) while that in venules, was 0.027 micrometers . s-1 . mmHg-1 (2.02 X 10(-10)m . s-1 . Pa-1). 4. In both capillaries and venules, filtration coefficient increased with decreasing pressure. 5. The difference between directly measured venular pressure and that calculated from the occlusion data was used to determine the contribution of the interstitium to fluid exchange. In the mesentery superfused with Krebs solution the tissue pressure so determined was found to be zero or subatmospheric initially but became increasingly positive with lengthening exposure of the mesentery.

Animals↗

Direct quantitation and detection of salmonellae in biological samples without enrichment, using two-step filtration and real-time PCR.

A new two-step filtration protocol followed by a real-time PCR assay based on SYBR green I detection was developed to directly quantitate salmonellae in two types of biological samples: i.e., chicken rinse and spent irrigation water. Four prefiltration filters, one type of final filter, and six protocols for recovery of salmonellae from the final filter were evaluated to identify an effective filtration protocol. This method was then combined with a real-time PCR assay based on detection of the invA gene. The best results were obtained by subsequent filtration of 100 ml of chicken rinse or 100 ml of spent irrigation water through filters with pore diameters of >40 mum to remove large particles and of 0.22 microm to recover the Salmonella cells. After this, the Salmonella cells were removed from the filter by vortexing in 1 ml of physiological saline, and this sample was then subjected to real-time quantitative PCR. The whole procedure could be completed within 3 h from sampling to quantitation, and cell numbers as low as 7.5 x 10(2) CFU per 100-ml sample could be quantified. Below this limit, qualitative detection of concentrations as low as 2.2 CFU/100 ml sample was possible on occasion. This study has contributed to the development of a simple, rapid, and reliable method for quantitation of salmonellae in food without the need for sample enrichment or DNA extraction.

Animals↗

Measurement of biocolloid collision efficiencies for granular activated carbon by use of a two-layer filtration model.

Point-of-use filters containing granular activated carbon (GAC) are an effective method for removing certain chemicals from water, but their ability to remove bacteria and viruses has been relatively untested. Collision efficiencies (alpha) were determined using clean-bed filtration theory for two bacteria (Raoutella terrigena 33257 and Escherichia coli 25922), a bacteriophage (MS2), and latex microspheres for four GAC samples. These GAC samples had particle size distributions that were bimodal, but only a single particle diameter can be used in the filtration equation. Therefore, consistent with previous reports, we used a particle diameter based on the smallest diameter of the particles (derived from the projected areas of 10% of the smallest particles). The bacterial collision efficiencies calculated using the filtration model were high (0.8 < or = alpha < or = 4.9), indicating that GAC was an effective capture material. Collision efficiencies greater than unity reflect an underestimation of the collision frequency, likely as a result of particle roughness and wide GAC size distributions. The collision efficiencies for microspheres (0.7 < or = alpha < or = 3.5) were similar to those obtained for bacteria, suggesting that the microspheres were a reasonable surrogate for the bacteria. The bacteriophage collision efficiencies ranged from > or = 0.2 to < or = 0.4. The predicted levels of removal for 1-cm-thick carbon beds ranged from 0.8 to 3 log for the bacteria and from 0.3 to 1.0 log for the phage. These tests demonstrated that GAC can be an effective material for removal of bacteria and phage and that GAC particle size is a more important factor than relative stickiness for effective particle removal.

Bacterial Adhesion↗

Precision and accuracy of recovery of Legionella pneumophila from seeded tap water by filtration and centrifugation.

Determination of the concentration of Legionella pneumophila in environmental water sites may be useful for the prediction of the risk of a particular site's causing Legionnaires' disease as well as for experimental studies of environmental growth or remediation. The precision and accuracy of recovery of two different L. pneumophila strains from seeded tap water samples were studied, with either filtration or centrifugation used to concentrate the bacteria. L. pneumophila grown on BCYE alpha agar or in Acanthamoeba castellanii was used to seed sterile tap water. Water samples were then either filtered (0.2-microns pore size) or centrifuged. An average of 53% (95% confidence interval [CI], 47 to 58%; n = 45) of the seeded L. pneumophila organisms were recovered by filtration with flat polycarbonate membranes. This recovery was significantly higher (P < 0.01) than that obtained by filtration with cast membranes (mean, 13%; 95% CI, 11 to 38%; n = 4) or by centrifugation at 3,800 x g for 30 min (mean, 14%; 95% CI, 2 to 25%; n = 9) or at 8,150 x g for 15 min (mean, 32%; 95% CI, 28 to 36%; n = 19). Recovery of L. pneumophila was not significantly different whether the bacteria were grown on plates or in amoebae. Use of a selective medium did not decrease the recovery efficiency, but preplating acid treatment of specimens caused an approximately 30% bacterial loss.(ABSTRACT TRUNCATED AT 250 WORDS)

Centrifugation↗

Membrane filtration of the Reiter treponeme.

A membrane filtration technique with commercially available membrane filters (Millipore Corp.) was effective for the removal of Reiter treponemes from liquids such as fluorescent-antibody conjugates, to which the organisms are added for adsorption. Reiter treponemes from an 8-day culture were not microscopically detectable in filtrates through membranes with a pore diameter of 0.45 mum, but treponemes were demonstrated in the filtrate by cultural methods. No organisms of the 8-day culture passed through a membrane filter having a pore size of 0.22 mum, as determined by microscopy and culture. Culture data indicated that a filter with a pore size of 0.1 mum was necessary to prevent passage of treponemes from 4-day cultures. It is recommended that a membrane filter with a pore size of 0.22 mum or smaller be used for the removal of Reiter treponemes from suspensions and that the age of the culture be considered in choosing filter pore size.

Culture Media↗

Rapid detection of simulated bacteremia by centrifugation and filtration.

A centrifugation-filtration procedure was developed to expedite the recovery of microorganisms from blood. Fresh whole human blood was inoculated with various aerobic and facultatively anaerobic microorganisms (3 to 18 per ml). The seeded blood was carefully overlaid on a Ficoll-Hypaque gradient (density, 1.114 g/ml) and centrifuged (400 x g) for 45 min at ambient temperature. The entire gradient (plasma, leukocytes, and Ficoll-Hypaque) was removed and filtered through a 0.22-micrometer membrane filter. The filters were then placed on chocolate agar and incubated at 35 degrees C in humidified air containing 5% CO2. No statistically significant differences were detected between the numbers of microorganisms recovered by filtration and by direct culture of the original inoculum. Most microorganisms were detected within 18 h after filtration. This system has excellent sensitivity and negligible toxicity.

Blood↗

Comparison of lysis-centrifugation with lysis-filtration and a conventional unvented bottle for blood cultures.

Evaluation of a commercially available lysis-centrifugation blood culture system (Isolator, DuPont Co., Wilmington, Del.) and a lysis-filtration blood culture system for 3,111 cultures showed that both methods had comparable recoveries (73 and 68%, respectively) of significant aerobic and facultatively anaerobic isolates. The unvented conventional blood culture bottle had a recovery rate of 59%. Although the lysis-centrifugation and lysis-filtration systems had comparable recoveries of pathogens, the lysis-centrifugation system had the advantage of having colonies immediately available for further testing. The contamination rate with the lysis-centrifugation system was 3%, compared with 6% with the lysis-filtration system and 0.4% with brain heart infusion.

Bacteria↗

Modified selective medium for isolation of Campylobacter spp. from feces: comparison with Preston medium, a blood-free medium, and a filtration system.

Our previously described (H. Goossens, M. De Boeck, and J. P. Butzler, Eur. J. Clin. Microbiol. 2:389-393, 1983) selective medium, consisting of cefoperazone (15 mg/liter), rifampin (10 mg/liter), colistin (10,000 IU/liter), and amphotericin B (2 mg/liter) (medium M1), for the isolation of Campylobacter jejuni and Campylobacter coli from stool specimens was modified as follows: cefoperazone (30 mg/liter), rifampin (10 mg/liter), and amphotericin B (2 mg/liter) (medium M2). A comparative study of the isolation of Campylobacter spp. from stool specimens was carried out with medium M1; medium M2; a selective blood-free medium consisting (per liter) of charcoal (4 g), ferrous sulfate (0.25 g), sodium pyruvate (0.25 g), casein hydrolysate (3 g), sodium deoxycholate (1 g), nutrient broth no. 2 (25 g), agar (12 g), and cefoperazone (32 mg) (medium M3); and Preston medium containing (per liter) trimethoprim (10 mg), rifampin (10 mg), polymyxin B (5,000 IU), and cycloheximide (100 mg) (medium M4). We also included a filtration system in which membrane filters were applied directly to the surface of the nonselective blood-free medium distributed in small petri dishes. A total of 5,276 stool specimens were tested: 2,788 stool specimens were tested on M1 and M3 in study 1; 2,488 stool specimens were inoculated on the four selective media in study 2, and the last 986 specimens of the 2,488 were tested in parallel with the filtration system. In study 2, 128 Campylobacter strains were isolated from 126 different patients; 85.0, 88.3, 82.5, and 66.6% of these strains were isolated on M1, M2, M3, and M4, respectively. No contaminating fecal flora was found on 65.4, 70.7, 62.4, and 40.3% of the M1, M2, M3, and M4 plates, respectively. Furthermore, C. coli was found to be more susceptible to antibiotics present in the selective media, particularly colistin and polymyxin B, than was C. jejuni. We therefore recommend M2 for the isolation of Campylobacter spp. Finally, the filtration method was found to be easy and cheap; although the sensitivity was low, this method allowed the isolation of new Campylobacter spp. which seem to be associated with diarrhea.

Adult↗

Clinical limitations of the estimation of glomerular filtration rate from height/plasma creatinine ratio: a comparison with simultaneous 51Cr edetic acid slope clearance.

A retrospective comparison of single determinations of glomerular filtration rate measured by 51chromium-edetic acid slope clearance (CEDTA) and height/plasma creatinine ratio (Ht/PC) was undertaken in 199 children aged between 3 and 16 years. Analysis of the data indicated that only if Ht/PC less than or equal to 1.2 cm/mumol per 1 was the relationship between Ht/PC and CEDTA linear. Over this range where renal function is significantly impaired the confidence limits for individual prediction of glomerular filtration rate were so wide that a definitive measurement of renal function would be necessary in clinical practice. Where Ht/PC greater than or equal to 1.2 the relationship was non-linear and the predictive confidence limits very wide. To apply the technique as a screening test for normal renal function (CEDTA greater than or equal to 80 ml/min per 1.73 m2) the Ht/PC would need to exceed 2.16. In our study this would have detected 57 out of 131 patients who had normal glomerular filtration rates and erroneously included 2 out of 68 with subnormal renal function.

Adolescent↗