Evaluation of inositol phosphates in urine after topical administration of myo-inositol hexaphosphate to female Wistar rats
F. GrásesAntonia Costa‐BauzáFrancisco BergaAdrián RodríguezRosa M. GomilaGabriel MartorellMelanie Raquel Martínez-Cignoni
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An inositol-requiring strain of Neurospora crassa was labelled during growth in liquid medium with [3H]inositol, and the levels of inositol phosphates and phosphoinositides were determined under inositol-sufficient and inositol-starved conditions. Because the mutant has an absolute requirement for inositol, the total mass of inositol-containing compounds could be determined. Inositol-containing lipids were identified by deacylation and co-migration with standards on h.p.l.c.; PtdIns3P, PtdIns4P, and PtdIns(4,5)P2 were found in approximately equal amounts, in addition to large amounts of PtdIns. Inositol starvation decreased the level of PtdIns to 10% of the sufficient level, and decreased the levels of the other phosphoinositides to about 25%. A number of inositol phosphates were found, including several InsP3s, InsP4s and InsP5s and phytic acid. Ins(1,4,5)P3 was identified by co-migration with standards on h.p.l.c. and by digestion with inositol phosphomonoesterase. High concentrations of all inositol phosphates were found in the extracellular medium in inositol-starved cultures. Inositol starvation on both liquid and solid agar media decreased the intracellular levels of some inositol phosphates, but increased the levels of phytic acid and several other inositol phosphates which may be its precursors and/or breakdown products. These results may indicate that inositol starvation induces phytic acid synthesis as a protection against the free-radical production and lipid peroxidation characteristic of inositol-less death.
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A study was made of quantitative urine cultures in bacteremic patients with urinary tract infection as the only identifiable source. These patients represent the most serious form of urinary tract infection and should be recognized as significant by the criteria used for interpreting quantitative urine cultures. In 83 patients, 15 (18%) had quantitative urine counts less than 10(5) colony-forming units (CFU)/mL. Ten (12%) had counts between 10(4) and 10(5) CFU/mL. The incidence of patients with less than 10(4) CFU/mL (5%) is probably a low estimate because of the urine culture methods used, and this requires further study. The results of this study support the view that significant urinary infections may be associated with quantitative urine counts of less than 10(5) CFU/mL.
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Evidence is presented to show that acid extracts of avian erythrocytes prelabelled for 24-48 h with myo-[3H]inositol contain the following myo-[3H]inositol trisphosphates (expressed as a percentage of total myo-[3H]inositol trisphosphates extracted): 36% myo-[3H]inositol 1,4,5-trisphosphate; 33.7% myo-[3H]inositol 1,3,4-trisphosphate; 13% myo-[3H]inositol 3,4,5-trisphosphate; 9.7% myo-[3H]inositol 3,4,6-trisphosphate; 4.4% myo-[3H]inositol 1,4,6-trisphosphate and 3.3% myo-[3H]inositol 1,3,6-trisphosphate. The only phosphatidyl-myo-[3H]inositol bisphosphate that could be detected in [3H]Ins-prelabelled avian erythrocytes was phosphatidyl-myo-[3H]inositol 4,5-bisphosphate. Cellular myo-[3H]inositol 3,4,5-trisphosphate may be synthesized by dephosphorylation of myo-[3H]inositol 3,4,5,6-tetrakisphosphate. D- and L-myo-[3H]inositol 1,4,6-trisphosphate and D- and L-myo-[3H]inositol 1,3,6-trisphosphate may be dephosphorylation products of myo-[3H]inositol 1,3,4,6-tetrakisphosphate.
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Objective Observation has led us to believe that urinary tract infections (UTI) may be difficult to diagnose from a dilute urine specimen. We conducted this study to determine the effect of urine concentration on identifying UTI based on urinalysis (UA) results. Methods We reviewed the UA results of febrile children under 36 months of age with positive urine cultures. We …
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When myo-[3H]inositol-prelabelled primary-cultured murine bone-marrow-derived macrophages were challenged with platelet-activating factor (PAF; 200 ng/ml), there was a rapid (2.5-fold at 10 s) rise in the intracellular concentration of D-myo-[3H]inositol 1,4,5-trisphosphate, followed by a rise in myo-[3H]inositol tetrakisphosphate. myo-[3H]Inositol tetrakisphosphate fractions were isolated by high-performance anion-exchange chromatography from myo-[3H]inositol-prelabelled chick erythrocytes and primary-cultured macrophages. In both cases [3H]iditol and [3H]inositol were the only significant products (greater than 90% of recovered radioactivity) after oxidation to completion with periodic acid, reduction with NaBH4 and dephosphorylation with alkaline phosphatase. The presence of [3H]inositol after this procedure is consistent with the occurrence of [3H]inositol 1,3,4,5-tetrakisphosphate in the cell extracts, whereas [3H]iditol could only be derived from D- or L-inositol 1,4,5,6-tetrakisphosphate. When [3H]inositol tetrakisphosphate fractions obtained from (A) unstimulated macrophages, (B) macrophages that had been stimulated with PAF for 40s or (C) chick erythrocytes were subjected to the above procedure, radioactivity was recovered in these polyols in the following proportions: A, 60-90% in iditol, with 10-40% in inositol; B, total radioactivity increased by a factor of 9.8, 94% being recovered in inositol and 8% in iditol; C, 70-80% in iditol and 20-30% in inositol. [3H]Iditol derived from myo-[3H]inositol tetrakisphosphate fractions from macrophages and chick erythrocytes was oxidized to sorbose by L-iditol dehydrogenase (L-iditol:NAD+2-oxidoreductase, 1.1.1.14) at the same rate as authentic L-iditol. D-[14C]Iditol, derived from D-myo-inositol 1,4,5-trisphosphate, was not oxidized by L-iditol dehydrogenase. This result indicates that the [3H]iditol was derived from L-myo-inositol inositol 1,4,5,6-tetrakisphosphate. The data are consistent with rapid PAF-sensitive synthesis of D-myo-[3H]inositol 1,3,4,5-tetrakisphosphate in macrophages, and demonstrate that L-myo-inositol 1,4,5,6-tetrakisphosphate is synthesized in both mammalian and avian cells. The levels of L-myo-[3H]inositol 1,4,5,6-tetrakisphosphate in primary-cultured macrophages are not acutely sensitive to PAF.
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Urinary tract infection (UTI) is defined as the presence of bacteria in urine along with symptoms of infection.UTIs occurs in 1.1% of girls and 1.4% of boys in the first year of life. The aim of the study was to assessthe usefulness of measurement of pro-inflammatory interleukin (IL)-6 and IL -8 concentrations in the urineand serum of children with UTI. A total of Eighty serum sample and seventy tow urine sample have beencollected from children with urinary tract infection, Their age ranged between 33days- 12 years old, fifty sixchildren of the same age collected as controls .Urine and serum IL-6 and IL-8 concentrations of both groupswere measured and compared. Urine and serum concentrations of IL -8 were significantly higher in childrenwith UTI compared with controls group (P = 0.0001, P = 0.0002) respectively, while there was no significantdifferent in urine and serum concentration of IL-6 of children with UTI and controls group(P = 0.1199 , P=0.572) respectively. The results demonstrate that IL-8 is a good biomarker for urinary tract infection, whileIL-6 is not.
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