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PubMed1984

A simple inulin assay for renal clearance determination using an immobilized beta-fructofuranosidase.

Current hospital practice for testing renal function is to use the creatinine clearance test. Inulin clearance, an inherently more accurate procedure, currently is only carried out by specialized laboratories because there is not a simple biochemical assay for inulin, that is, an assay that could be carried out by any laboratory without special facilities. We have developed a simple enzymatic assay system for measuring inulin in plasma and urine. The procedure uses a beta-fructofuranosidase immobilized on Concanavalin A to convert inulin to fructose. Fructose is then measured by measuring the NADH----NAD conversion produced when fructose is converted to sorbitol by the enzyme sorbitol dehydrogenase. Kinetic parameters, binding capacities, and operating conditions for the immobilized beta-fructofuranosidase were determined as well as general operating parameters for the complete assay system. This system offers the potential for replacing the creatinine clearance test as the assay of choice for renal function.

Enzymes, Immobilized↗

PubMed1991

Distribution of subcutaneously administered inulin between blood and peripheral lymph in the rabbit.

In experiments on rabbit peripheral lymph, the contribution of the blood and lymphatic system to the whole-body distribution of inulin after subcutaneous administration has been investigated and the effects of hyaluronidase and of thermal stimulus at the administration site examined. Inulin concentrations in lymph exceeded plasma concentrations by more than 100-fold. At the end of the experiment (90 min) the amount of drug in the total lymph collected was about one-seventh the amount found in urine. The blood system, as a result of higher circulation at the administration site distributes inulin from the subcutis more rapidly than does lymph. Hyaluronidase did not influence inulin concentrations in blood and lymph but thermal stimulus significantly decreased both concentration and total distribution. The decrease resulted from a developed oedema and vasoconstriction in the skin and subcutis of the cannulated extremity.

Animals↗

PubMed1988

In vivo permeability of the human placenta to inulin and mannitol.

1. Permeability of the term human placenta to inulin and mannitol was measured in six normal pregnancies. 2. Bolus and continuous infusion of inulin and mannitol to the mother for 1-2 h before delivery by Caesarean section achieved steady-state maternal serum concentrations. Net accumulation of inulin and mannitol in the fetus was measured by the subsequent neonatal urinary excretion. 3. Transplacental clearance was calculated by dividing the net flux by the time-averaged transplacental concentration gradient. Clearance for inulin was 0.86 +/- 0.1 ml min-1 (mean +/- S.E.M.) and for mannitol 7.3 +/- 0.8 ml min-1 (n = 6 in both cases).

Female↗

PubMed1976

Early postglomerular plasma concentrations of chloride, sodium, and inulin in the rat kidney.

Sodium, chloride, and inulin concentrations were measured in plasma collected from the terminal portions of long efferent vessels at the subcapsular surface of the rat kidney. Sodium concentration equaled and the concentrations of chloride and inulin were less than those in peripheral plasma. During benzolamide infusion, chloride concentration equaled while inulin concentration remained less than in peripheral plasma. In free-flow micropuncture samples collected randomly during control conditions, chloride concentration rose rapidly in the early proximal tubule and then remained elevated and constant throughout the remainder of the proximal tubule accessible to micropuncture. These experiments indicate that normally tubular reabsorbate low in chloride and inulin is added to the blood traversing the early postglomerular vessels before reaching the kidney surface. Bases on the analyses of proximal tubular fluid, this type of reabsorbate appears available only from the early proximal tubular segment. We conclude that a close functional relationship exists between the first segment of the proximal tubule and the early postglomerular blood supply characteristic of the superficial cortical nephron.

Animals↗

PubMed1985

Measurement of glomerular filtration rate in conscious unrestrained rats with inulin infused by implanted osmotic pumps.

A method is described for determining glomerular filtration rate (GFR) in conscious unrestrained rats without extracellular volume expansion. The glomerular marker used was 14C-labeled inulin infused in the minimal fluid volume of 1 microliter/h by intraperitoneally implanted osmotic minipumps. The workability, reproducibility, and precision of the technique was evaluated in sham-operated (sham) and uninephrectomized (UNI-NX) rats for 6 days. In six sham-operated rats the clearance of [14C]-inulin calculated from the plasma value obtained in a blood sample taken at 10 A.M. before food consumption was (means +/- SE): day 1 after pump implantation: 1.38 +/- 0.05; day 2: 1.27 +/- 0.05; day 3: 1.46 +/- 0.16; day 4: 1.45 +/- 0.15; day 5: 1.33 +/- 0.08; day 6: 1.38 +/- 0.11 ml/min. In six UNI-NX rats the corresponding values were: day 1: 0.88 +/- 0.04; day 2: 0.79 +/- 0.05; day 3: 0.91 +/- 0.03; day 4: 0.91 +/- 0.03; day 5: 0.83 +/- 0.06; day 6: 0.87 +/- 0.05 ml/min. When determined on day 2, 3, or 6 at 6 P.M., i.e., at the end of the food consumption period, [14C]inulin clearance was increased in all animals compared with the value determined at 10 A.M. in the fasted state. The use of implanted osmotic minipumps for delivering a glomerular marker such as [14C]inulin allows the determination of GFR in conscious unrestrained rats with normal fluid balance conditions. This method appears to be particularly appropriate for studying the influence of the intake and composition of food on GFR in physiological conditions.

Animals↗

PubMed1995

Inulin as a marker of dilution of bronchoalveolar lavage in asthmatic and normal subjects.

Bronchoalveolar lavage (BAL) fluid is a variable mixture of instilled and lung fluid, which makes interpretation of solute concentrations difficult. We describe the use of inulin as a marker of dilution of BAL in human subjects. BAL, using saline containing 0.1 mM inulin, was safely performed in 13 subjects with mild asthma and 11 normal subjects. The dilution factor (DF: inulin concentration in BAL fluid/inulin concentration in instilled fluid) was measured spectrophotometrically, and it was used to calculate the volume of lung fluid in BAL fluid. There was no significant difference between the median (range) DF of 0.931 (0.825 to 0.952) in asthmatics and 0.907 (0.768 to 0.985) in control subjects (p = 0.77). There was wide individual variation in, but no significant difference between, the lung fluid volume of 8.1 ml (5.4 to 22.2) in asthmatics and 12.3 ml (1.9 to 30.6) in control subjects (p = 0.56), thus validating comparisons of concentrations per ml of BAL fluid. Alternatively, concentrations can be compared per ml of lung fluid. Inulin fulfilled the requirements for a marker of dilution of BAL, enabling the validation and standardization of comparisons of solute concentrations in BAL fluid.

Adult↗

PubMed1984

Analysis of contact-rehydration in terrestrial gastropods: absorption of 14C-inulin through the epithelium of the foot.

Contact-rehydration in terrestrial slugs involves a specific drinking behaviour during which water is rapidly absorbed through the integument of the foot. When dehydrated slugs were placed on wet filter paper containing 14C-inulin, they displayed the characteristic drinking posture and absorbed both water and 14C-inulin. Samples of haemolymph from dehydrated slugs after 12 min of contact-rehydration contained about 6 micrograms of 14C-inulin 100 mg-1 of haemolymph (0.24 mmol l-1 14C-inulin in the substrate). The haemolymph of hydrated slugs however contained no detectable radioactivity after 12 min on the filter paper. Electron microscopy revealed that the intercellular spaces between the epithelial cells of the foot were reduced in dehydrated slugs, but were rapidly enlarged during contact-rehydration. It is concluded that contact-rehydration in terrestrial slugs is mediated by bulk flow of water through an epithelial paracellular pathway in the integument of the foot.

Animals↗

PubMed1998

Evaluation of single-injection method of inulin and creatinine as a renal function test in normal cats.

A single injection method for estimating glomerular filtration rate (GFR) by measuring plasma inulin and creatinine clearances was evaluated in 10 healthy cats. GFRs were estimated from the plasma clearance (PC) by dividing the injected dose of an indicator by the area under the plasma disappearance curve (AUC). AUC was determined by 2 common pharmacokinetic analyses, the two-compartment model and the trapezoidal rule. AUCs determined by these two methods were significantly correlated both in inulin (r = 0.993) and creatinine (r = 0.959). To minimize errors, GFR was estimated by PC only if AUC/10 was greater than the area under the curve from the final sampling time to infinitive (A2). GFRs determined by PC of inulin at final sampling time of 180 and 240 min were 3.61 +/- 0.64 and 3.63 +/- 0.67 ml/min/kg of body weight (mean +/- SD), respectively. These values corresponded to the reference range reported for normal cats. In contrast, when creatinine was used as a maker, A2 was always greater than AUC/10 at any final sampling time and GFRs estimated using these AUCs of creatinine were significantly greater than those of inulin, suggesting creatinine may not be suitable indicator for the single injection method.

Animals↗

PubMed2001

Evaluation of plasma clearance of inulin in clinically normal and partially nephrectomized cats.

OBJECTIVE: To evaluate accuracy of measuring plasma clearance of inulin as an alternative renal function test for estimation of glomerular filtration rate (GFR) in cats. ANIMALS: 10 cats, first studied with intact kidneys and subsequently studied following partial nephrectomy. PROCEDURE: Clearance studies were performed in 10 clinically normal cats; those same cats then underwent partial nephrectomy, and clearance studies were performed again. Plasma concentration of inulin was determined after administration at 50 mg/kg of body weight to cats while renally intact and 45 mg/kg after the partial nephrectomy. Plasma clearance of inulin (PCin) was determined by dividing the dose by the area under the plasma inulin concentration versus time curve. Results for PCin were compared with values obtained simultaneously for urinary clearance of exogenously administered creatinine (Ccr), a widely accepted method for measurement of GFR in cats with intact kidneys and cats with reduced renal mass. RESULTS: Results of PCin were strongly correlated (r2 = 0.912, P < 0.001) with Ccr. Repeatability of determination of PCin was similar to that of Ccr. Sensitivity and specificity of PCin were superior and equivalent to that of Ccr, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Determination of PCin provides a reliable estimate of GFR in cats and is a promising alternative to determining Ccr in cats.

Animals↗

PubMed1982

Inulin uptake and washout in contracting and quiescent rat papillary muscle.

Equilibration and washout curves of inulin were determined in isolated rat papillary muscles. Inulin uptake did not reach a steady-state value even after a long equilibration time. The effects of contractile activity were investigated. Isometric contractions increased the rate of inulin uptake and washout. In isotonic conditions, a smaller increase in the uptake and washout was observed. These changes could be attributed to an actual increase of inulin distribution space and/or to an accelerated diffusion kinetics.

Animals↗

PubMed2002

Effect of honey on the growth of and acid production by human intestinal Bifidobacterium spp.: an in vitro comparison with commercial oligosaccharides and inulin.

Five human intestinal Bifidobacterium spp., B. longum, B. adolescentis, B. breve, B. bifidum, and B. infantis, were cultured in reinforced clostridial medium (control) and in reinforced clostridial medium supplemented with 5% (wt/vol) honey, fructooligosaccharide (FOS), galactooligosaccharide (GOS), and inulin. Inoculated samples were incubated anaerobically at 37degrees C for 48 h. Samples were collected at 12-h intervals and examined for specific growth rate. Levels of fermentation end products (lactic and acetic acids) were measured by high-pressure liquid chromatography. Honey enhanced the growth of the five cultures much like FOS, GOS, and inulin did. Honey, FOS, GOS, and inulin were especially effective (P < 0.05) in sustaining the growth of these cultures after 24 h of incubation as compared with the control treatment. Overall, the effects of honey on lactic and acetic acid production by intestinal Bifidobacterium spp. were similar to those of FOS, GOS, and inulin.

Acetic Acid↗

PubMedDate not supplied

Inhibitory effect of dietary inulin or oligofructose on the development of cancer metastases.

The possible influence of continuous dietary treatment with 15% inulin or oligofructose on the development of lung metastases of a transplantable liver tumor in young male C3H mice was investigated. Microscopical examination demonstrated a distinct inhibitory effect of this dietary treatment on the development of lung metastases of this tumor. There were 59% of mice bearing lung metastases in the control group, 36% in the inulin fed group and 35% in the oligofructose fed group. The total number of lung metastases was 37 in the control group, 18 in inulin fed and 6 in oligofructose fed mice. Several possible, mechanisms hypothetically involved in this astonishing inhibition of the development of lung metastases by dietary treatment with inulin or oligofructose are discussed.

Animals↗

PubMedDate not supplied

Non-toxic potentiation of cancer radiotherapy by dietary oligofructose or inulin.

Non-toxic, dietary treatment with oligofructose or inulin clearly inhibited the growth of a transplantable mouse liver tumor (TLT) and potentiated its chemotherapy. Thus, it appeared interesting to investigate the possible radiotherapy-potentiating effects of the same dietary treatment. Dietary treatment with 15% oligofructose or inulin incorporated in the basal diet was started four weeks before intramuscular transplantation of TLT tumor cells into young adult male mice of the NMRI strain and was continued until the end of the experiment. When the tumors reached approximately 1000 mm3 they were irradiated with a single X-ray dose of 5 to 20 Gy. Tumor dimensions were measured twice weekly and their mean volume per group of mice was compared to the control groups fed the basal diet. This non-toxic dietary treatment with oligofructose or inulin potentiated the effects of radiotherapy at an optimal dose of 10 Gy to a statistically very highly significant (p < 0.0001) level. They were similar for oligofructose and inulin. The introduction of such non-toxic adjuvant treatment potentiating the effect of cancer radiotherapy in classical protocols of human cancer treatment appears to be possible and without any additional risk for the patients.

Animals↗

PubMed2004

High-performance anion-exchange chromatography coupled with pulsed amperometric detection and capillary zone electrophoresis with indirect ultra violet detection as powerful tools to evaluate prebiotic properties of fructooligosaccharides and inulin.

Fructooligosaccharides (FOS) and inulin are food grade non-digestible carbohydrates that exert beneficial nutritional effect. This paper describes the suitability of high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) and capillary zone electrophoresis (CZE) to evaluate fermentation properties of FOS and inulin in pure Bifidobacterium cultures; and to study their effects on faecal cultures (microbial population and short-chain fatty acids). Prebiotic effectiveness of FOS and inulin of different degrees of polymerization was evaluated monitoring the changes in their molecular weight distribution during the in vitro growth of selected Bifidobacterium strains. The qualitative analysis of the residual soluble oligosaccharides or polysaccharides from Raftilose Synergy, Raftiline HP and Raftilose P95 was carried out by HPAEC-PAD, using a CarboPac PA 100 column and an appositely optimized gradient elution program. Under the optimized gradient elution conditions, glucose, fructose, sucrose were resolved from each other and from fructans with a DP ranging from 3 (1-kestose) to 60. The chromatographic profiles of the spent broths pointed out that almost every strain presented a different capability to ferment fructan chains of variable DP, indicating wide strain to strain differences. To explore the prebiotic effect of FOS and inulin, related to of short chain fatty acids (SCFAs) accumulation in faecal cultures due to fermentative metabolism of intestinal microflora, analysis of SCFAs, acetic and lactic acid was achieved by co-electroosmotic capillary electrophoresis, where the electrophoretic mobility of the anionic analytes and electroosmotic flow (EOF) were similarly directed. Moreover, the use of UV detection for the analyses of our organic anions required a running electrolyte which allowed indirect detection. The optimization of the capillary electrophoretic conditions was carried out by applying a chemometric study based on the use of the experimental design, the effects of three parameters, i.e. temperature, voltage and percentage of methanol added to the background electrolyte were investigated.

Anion Exchange Resins↗

PubMed2005

[Estimation of glomerular filtration rate by inulin clearance: comparison with creatinine clearance].

Inulin clearance (Cin) is widely believed to be the gold standard of the glomerular filtration rate (GFR). However, in Japan, Cin has not been officially recognized by the Ministry of Health, Labour and Welfare of Japan for clinical use. Creatinine clearance (Ccr) has been used to estimate the renal function of patients, but there have been many studies in which Ccr estimates were GFR falsely high because the metabolism and tubular excretion of creatinine widely varied according to the pathophysiological state of the patient. In the present study, we determined Cin and Ccr simultaneously in 116 adult patients with renal diseases and diabetic mellitus. The clearance study was performed by the modified Wesson's method. The inulin preparation was FFI-1010 (Fuji Yakuhin Co. Ltd.). Inulin in serum and urine was determined by the newly devised enzymatic assay (Toyobo Co. Ltd.), which is specific for inulin. The mean Cin was 35.0 +/- 14.4 ml/min/1.73 m2. The mean Ccr(the enzyme assay) was 63.6 +/- 24.1 ml/min/1.73 m2 and that of the kinetic Jaffe assay was 55.3 +/- 19.3 ml/min/1.73 m2. Mean Ccr/Cin was 1.93 +/- 0.73, 1.69 +/- 0.62, respectively. This ratio was significantly different(p < 0.05) in the degree of reduction of Cin, with values of 2.07 +/- 0.82 (Cin < 40 ml/min/1.73 m2) and 1.64 +/- 0.32(40 < Cin < 80 ml/min/1.73 m2), respectively. Only 8 patients were classified into the same degree of reduced renal function (the Guideline of Japanese Society of Nephrology). The findings of this study suggest that the GFR determined by Ccr could misjudge the renal function of patient and delay the administration of proper treatment of the patient. Introduction of Cin into the clinical field is necessary to avoid this delay.

Acute Disease↗

PubMed1990

Determination of excretion of inulin, creatinine, sodium sulfanilate, and phenolsulfonphthalein to assess renal function in goats.

Excretion of creatinine, sodium sulfanilate (SS), and phenolsulfonphthalein (PSP) was studied in healthy goats. In conscious goats, mean (+/- SEM) inulin clearance was 2.26 +/- 0.08 ml/min/kg of body weight. Endogenous creatinine clearance, 1.97 +/- 0.09 ml/min/kg, underestimated inulin clearance (P less than 0.01), probably because of the presence of noncreatinine chromogens in caprine plasma. The estimated renal clearance of PSP was 6.88 +/- 0.39 ml/min/kg, whereas the estimated renal clearance of SS was 3.71 +/- 0.39 ml/min/kg. Both exceeded inulin clearance (P less than 0.01), confirming renal tubular secretion of both compounds. In 6 anesthetized goats, exogenous creatinine clearance and SS clearance exceeded inulin clearance (P less than 0.05). Results of stop-flow experiments documented secretion of creatinine and SS by the proximal portion of the caprine nephron. Plasma half-life of PSP in uninephrectomized goats exceeded that in intact goats (20.2 +/- 1.5 min vs 11.9 +/- 0.7 min; P less than 0.01). Similarly, plasma half-life of SS was greater in goats after uninephrectomy (58.2 +/- 6.2 min vs 30.4 +/- 1.2 min; P less than 0.01).

Animals↗

PubMed1986

Comparison between the renal clearance of endogenous creatinine and inulin in the sheep.

Simultaneous renal clearances of inulin and endogenous creatinine were determined during inulin infusions in sheep. The calculated regression of creatinine on inulin clearance differed significantly from the expected line. Discrepancies might be explained by absorption and secretion of creatinine under differing conditions, but it is suggested that lack of specificity of the method used for creatinine determination is an equally likely cause. As the difference between the two clearances was not great (within the range 14 to 48 ml min-1 it was not significant at P less than 0.05), creatinine clearance remains a useful indicator of glomerular filtration. For greatest accuracy, however, inulin clearance remains the method of choice.

Animals↗

PubMed1985

The permeability of normal, adenomatous, ulcerative colitic and malignant large bowel epithelial cell membranes to inulin.

We measured the permeability of normal, adenomatous, colitic and malignant large bowel epithelial cells by immersing fragments of large bowel mucosa in radiolabelled inulin and comparing autoradiograph grain density inside and outside cells after incubation. All the carcinomas studied showed extensive uptake of inulin within 5 min, while normal, adenomatous and colitic epithelial cells completely excluded inulin for 30 min. We found no difference in the proportion of epithelial cells incorporating uridine into RNA in carcinomatous and normal mucosa, and this suggests that the increased inulin permeability of carcinoma cell membranes was not due to leakage into non-viable cells. Experiments with cytochalasin B also showed that increased pinocytosis by carcinoma cells could not account for the difference. The relative impermeability of adenomatous and colitic cells suggests that increased permeability is not caused by increased proliferation. The consistent finding of increased permeability in the plasma membranes of carcinoma cells suggests that this may be more than an epiphenomenon of malignancy. It also suggests that measurement of cell permeability may have a role in distinguishing malignant from benign epithelial neoplasms.

Adenoma↗