Determinants of Brushite Stone Formation: A Case-Control Study

Purpose

The occurrence of brushite stones has increased during recent years. However, the pathogenic factors driving the development of brushite stones remain unclear.

Methods

Twenty-eight brushite stone formers and 28 age-, sex- and BMI-matched healthy individuals were enrolled in this case-control study. Anthropometric, clinical, 24 h urinary parameters and dietary intake from 7-day weighed food records were assessed.

Results

Pure brushite stones were present in 46% of patients, while calcium oxalate was the major secondary stone component. Urinary pH and oxalate excretion were significantly higher, whereas urinary citrate was lower in patients as compared to healthy controls. Despite lower dietary intake, urinary calcium excretion was significantly higher in brushite stone patients. Binary logistic regression analysis revealed pH>6.50 (OR 7.296; p = 0.035), calcium>6.40 mmol/24 h (OR 25.213; p = 0.001) and citrate excretion <2.600 mmol/24 h (OR 15.352; p = 0.005) as urinary risk factors for brushite stone formation. A total of 56% of patients exhibited distal renal tubular acidosis (dRTA). Urinary pH, calcium and citrate excretion did not significantly differ between patients with or without dRTA.

Conclusions

Hypercalciuria, a diminished citrate excretion and an elevated pH turned out to be the major urinary determinants of brushite stone formation. Interestingly, urinary phosphate was not associated with urolithiasis. The increased urinary oxalate excretion, possibly due to decreased calcium intake, promotes the risk of mixed stone formation with calcium oxalate. Neither dietary factors nor dRTA can account as cause for hypercalciuria, higher urinary pH and diminished citrate excretion. Further research is needed to define the role of dRTA in brushite stone formation and to evaluate the hypothesis of an acquired acidification defect.     

Expression of the xylulose 5-phosphate phosphoketolase gene, xpkA, from Lactobacillus pentosus MD363 is induced by sugars that are fermented via the phosphoketolase pathway and is repressed by glucose mediated by CcpA and the mannose phosphoenolpyruvate phosphotransferase system

Purification of xylulose 5-phosphate phosphoketolase (XpkA), the central enzyme of the phosphoketolase pathway (PKP) in lactic acid bacteria, and cloning and sequence analysis of the encoding gene, xpkA, from Lactobacillus pentosus MD363 are described. xpkA encodes a 788-amino-acid protein with a calculated mass of 88,705 Da. Expression of xpkA in Escherichia coli led to an increase in XpkA activity, while an xpkA knockout mutant of L. pentosus lost XpkA activity and was not able to grow on energy sources that are fermented via the PKP, indicating that xpkA encodes an enzyme with phosphoketolase activity. A database search revealed that there are high levels of similarity between XpkA and a phosphoketolase from Bifidobacterium lactis and between XpkA and a (putative) protein present in a number of evolutionarily distantly related organisms (up to 54% identical residues). Expression of xpkA in L. pentosus was induced by sugars that are fermented via the PKP and was repressed by glucose mediated by carbon catabolite protein A (CcpA) and by the mannose phosphoenolpyruvate phosphotransferase system. Most of the residues involved in correct binding of the cofactor thiamine pyrophosphate (TPP) that are conserved in transketolase, pyruvate decarboxylase, and pyruvate oxidase were also conserved at a similar position in XpkA, implying that there is a similar TPP-binding fold in XpkA.     

Comparison of the radiotoxicity of two alpha-particle-emitting immunoconjugates,terbium-149 and bismuth-213, directed against a tumor-specific,exon 9 deleted (d9) E-cadherin adhesion protein

We investigated the effects of the alpha-particle emitters (149)Tb and (213)Bi coupled to a tumor-specific antibody targeting the mutated delta 9 E-cadherin (d9 E-Cad) on single cells and cell pellets. The d9 mutation of the adhesion molecule E-cadherin is found in 10% of diffuse-type gastric cancers and is not expressed in normal tissue. Human breast cancer cells (MDA-MB-435S) transfected with d9 E-Cad or the wild-type E-cadherin gene were used to study the effects of anti-d9 E-Cad MAb coupled to (149)Tb and (213)Bi ((149)Tb-d9 MAb and (213)Bi-d9 MAb). The density of binding sites determined on transfected MDA tumor cells by Scatchard analysis and flow cytometry varied from 4 x 10(4) to 6 x 10(4) antigens per cell. Internalization of radioimmunoconjugates by cells expressing d9 E-Cad was less than 10% of bound antibody within 240 min. The effect of the radioimmunoconjugates on cell suspensions and cell pellets was quantified by [(3)H]thymidine incorporation, and the dose to the cell nuclei was determined using microdosimetric calculations. (149)Tb and (213)Bi immunoconjugates affected cells in suspension similarly. Significant differences in the proliferation capacity of d9 E-cadherin- and wild-type E-cadherin-expressing cells were observed at activity concentrations around 185 kBq/ml, corresponding to antibody concentrations between 200 ng/ml and 1000 ng/ml. Proliferation after incubation with (213)Bi-d9 MAb was 50% greater in pelleted wild-type E-Cad-expressing cells compared to wild-type E-Cad cells in suspension. In contrast, the proliferation of pelleted d9 E-Cad cells was similar to that of d9 E-Cad cells in suspension. For (149)Tb-d9 MAb, no significant difference was found between pelleted cells and cells in suspension for low activity concentrations. However, at high activity concentrations, (149)Tb-d9 MAb had only a small effect on pelleted cells. These in vitro studies demonstrate different effects of (149)Tb and (213)Bi conjugated to a tumor-specific antibody toward single cells and tumor cell pellets. Microdosimetric simulation of single cell survival after alpha-particle irradiation modeled the experimental results with reasonable accuracy.     

Associations between light-harvesting complexes and Photosystem II from Marchantia polymorpha L. determined by two- and three-dimensional electron microscopy

Assemblies of Photosystem II and light-harvesting proteins were purified from the liverwort Marchantia polymorpha and investigated by two- and three-dimensional transmission electron microscopy of negatively stained specimens. By single-particle analysis, it was determined that about 25% of the particles are rectangular or slightly S-shaped with dimensions of 285 A in length, 144 A in width, 84 A in height, while the membrane part is about 52 A thick. This structure reveals the same architecture as that of a Photosystem II-light-harvesting assembly from seed plants. An overlay of the projection structure of the liverwort's complex with a projection structure deduced from stained trimeric LHC II crystals from pea confirmed the locations of trimeric LHC II within the liverwort's complex. Remarkably tight associations of LHC II and other chlorophyll a/b binding proteins with the PS II core complex are observed. More than 50% of the Photosystem II particles from the liverwort carry one or two additional masses. These extra masses are found to consist of an additional LHC II trimer and probably a chlorophyll a/b binding protein. For the first time, a three-dimensional structure of such a large assembly is defined.     

The absence of nidogen 1 does not affect murine basement membrane formation

Nidogen 1 is a highly conserved protein in mammals, Drosophila melanogaster, Caenorhabditis elegans, and ascidians and is found in all basement membranes. It has been proposed that nidogen 1 connects the laminin and collagen IV networks, so stabilizing the basement membrane, and integrates other proteins, including perlecan, into the basement membrane. To define the role of nidogen 1 in basement membranes in vivo, we produced a null mutation of the NID-1 gene in embryonic stem cells and used these to derive mouse lines. Homozygous animals produce neither nidogen 1 mRNA nor protein. Surprisingly, they show no overt abnormalities and are fertile, their basement membrane structures appearing normal. Nidogen 2 staining is increased in certain basement membranes, where it is normally only found in scant amounts. This occurs by either redistribution from other extracellular matrices or unmasking of nidogen 2 epitopes, as its production does not appear to be upregulated. The results show that nidogen 1 is not required for basement membrane formation or maintenance.     

Zum Einflu? einer Autobahn im Bau und w?hrend des Betriebs auf die Brutbiologie von Kohlmeisen (Parus major) und Blaumeisen (P. caeruleus)

Zusammenfassung Im Februar 1992 wurde an der in Bau befindlichen Autobahn BAB 66 zwischen Steinau und Schlüchtern (5021'N, 0931'E) ein Nistkastenkontrollgebiet mit 310 Nistkästen eingerichtet, um die Auswirkungen der Autobahn vor und nach Inbetriebnahme auf die Brutbiologie von Meisen zu untersuchen. Als Vergleichsgebiete dienen ein direkt benachbartes sowie ein ca. 5 km entferntes Untersuchungsgebiet. Zum Vergleich der Gewichtsentwicklung der Nestlinge wurde ein drittes autobahnfernes Gebiet herangezogen.In einzelnen Jahren finden sich zwischen den Gebieten Unterschiede in der Zusammensetzung der Brutpopulation, der Besetzungsrate der Nistkästen, dem Legebeginn, der Gelegegröße, der Schlüpfrate oder im Bruterfolg. Diese Unterschiede treten in verschiedenen Jahren und in unterschiedlichen Gebieten auf, so daß sich keine generelle Abweichung des Gebietes an der Autobahn von den straßenfernen Gebieten feststellen läßt. Die Eröffnung der Autobahn hatte keinen Effekt auf die untersuchten Parameter.Die Gewichte der Nestlinge im Gebiet an der Autobahn waren im Jahr 1996 nur am 12. und 13. Nestlingstag signifikant geringer als die Gewichte der Nestlinge in einem autobahnfernen gleichartig strukturierten Biotop. Die Ausflugsgewichte der Jungvögel an der A66 liegen jedoch ca. 2 g höher als in städtischen Biotopen.Störungen durch den Straßenverkehr, wie sie von anderen Autoren berichtet werden, ließen sich in unserem Untersuchungsgebiet nicht feststellen. In diesem Zusammenhang werden die geringe Störungsempfindlichkeit der Kohl- und Blaumeisen sowie der bisher geringe Verkehr auf der Autobahn diskutiert.

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The impact of a motorway in construction and after opening to traffic on the breeding biology of Great Tit (Parus major) and Blue Tit (P. caeruleus)

Summary In February 1992, a study area (A 66) with 310 nestboxes was installed along the construction line of a four laned motorway between Steinau and Schlüchtern (5021'N, 0931'E). One area in the immediate neighbourhood (VG1) and one 5 km distant (VG2) from the study area at the motorway (see Fig. 1) served as controls for comparison. In all three areas, the nestboxes were checked weekly during the breeding season from the beginning of May until July. Species composition of the birds using nestboxes, rate of occupation of nestboxes, and, for Great Tits (Parus major) and Blue Tits (P. caeruleus), date of the first egg, clutch size, hatching rate and number of birds fledged were recorded. The study included 5 breeding seasons from 1992 to 1996, 3 years during the period of construction and 2 years after the opening of the motorway to traffic in December 1994. In 1996, the nestling weight of 20 broods of Great Tits at the A66 was recorded daily and compared with that of all nestlings in another study area (VG3, see Fig. 1).The data were analyzed separately for each year. Distributions of frequencies were analyzed with the x² test, the other data with the Kruskal-Wallis-Test (H-Test). In case of significant differences between the study areas, the Mann Whitney U-test was used to determine which study area differed from the others. Nestlings bodymass was compared using the t-test.The species composition was similar in all three areas (Fig. 2), and it did not change after the opening of the motorway to traffic. The rate of occupation of the nestboxes, dates of laying of the first egg, clutch size, hatching rate and breeding success vary between years and between study areas (Table 1 to 5); data that differ significantly from those of two other areas are indicated by bold print. Such differences were observed in various years and involved all three areas. In study area A66, clutch size of Great Tits was higher in 1993; hatching success of Great Tits was lower in 1994 and that of Blue Tits was lower in 1993. However, there was no general trend that area A66 at the motorway was different from the other areas, either before or after being opened to the traffic. The weight of nestlings in study area A66 at the motorway was similar to that in comparable area nearby; only on day 12 and 13 after hatching was it slightly lower (Tab. 6). The last measurement on day 15 indicates a nestling weight of 16.2 g for the area at the motorway, which is in normal for deciduous forest habitats and approximately 2 g higher than that recorded in urban habitats.In brief, the motorway did not affect the species composition, rate of occupied nestboxes or the various breeding parameters; in particular, the moving traffic after opening did not seem to have any effect. The breeding parameters from the study area at the motorway were typical for the Schlüchtern region. Density reductions in breeding populations, as discussed by several authors, or disturbances caused by moving traffic, as previously reported, were not observed in our study. However, our key species, Great Tits and Blue Tits, are not generally very sensitive to interferences. Also, the level of traffic on the motorway was still rather low. This must be considered when our findings are generalized.