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Phosphate (Pi) is an essential nutrient, and its plasma levels are under tight hormonal control. Uphill transport of Pi into cells is mediated by the two Na-dependent Pi transporter families SLC34 and SLC20. The molecular identity of a potential Pi export pathway is controversial, though XPR1 has recently been suggested by Giovannini and coworkers to mediate Pi export. We expressed XPR1 in Xenopus oocytes to determine its functional characteristics. Xenopus isoforms of proteins were used to avoid species incompatibility. Protein tagging confirmed the localization of XPR1 at the plasma membrane. Efflux experiments, however, failed to detect translocation of Pi attributable to XPR1. We tested various counter ions and export medium compositions (pH, plasma) as well as potential protein co-factors that could stimulate the activity of XPR1, though without success. Expression of truncated XPR1 constructs and individual domains of XPR1 (SPX, transmembrane core, C-terminus) demonstrated downregulation of the uptake of Pi mediated by the C-terminal domain of XPR1. Tethering the C-terminus to the transmembrane core changed the kinetics of the inhibition and the presence of the SPX domain blunted the inhibitory effect. Our observations suggest a regulatory role of XPR1 in cellular Pi handling rather than a function as Pi exporter. Accordingly, XPR1 senses intracellular Pi levels via its SPX domain and downregulates cellular Pi uptake via the C-terminal domain. The molecular identity of a potential Pi export protein remains therefore elusive. [ABSTRACT FROM AUTHOR]

Introduction: While the incidence of reverse total shoulder arthroplasty (rTSA) is increasing constantly, newer implants with designs other than the classic Grammont geometry are gaining importance. More anatomic inclination angles and lateralization are supposed to have a positive impact on clinical results and complication rates. Presentation of midterm results therefore is important to support these assumptions. The aim of this study was to report the midterm clinical outcome of primary rTSA with an uncemented humeral short-stem prosthesis (USSP) with a humeral inclination angle of 145° and the analysis of different variables on the outcome. Methods: This is a retrospective study of all patients with primary rTSA using an USSP and a combined humeral inclination angle of 145° (Ascend™ flex, Stryker) with a minimum clinical follow-up of 2 years. The implant combines a 132.5° inclination for the humeral stem with an additional 12.5° for the polyethylene inlay. Primary outcomes were patient-reported outcome measures: ASES score, simple shoulder test (SST) and subjective shoulder value (SSV). Secondary outcomes were complication and revision rates. We analyzed different variables: preoperatively gender, age, indication for surgery and status of rotator cuff. Also, the glenoid morphology was classified according to Walch and a proximal humerus cortical bone thickness measurement (CBT avg) of 6 mm was used as a threshold for osteoporosis. Postoperatively, we analyzed different radiologic parameters: filling ratio, distalization and lateralization angles according to Boutsiadis. Results: A total of 84 out of 99 (85%) patients with a mean FU of 46.7 months (range 24–80 months) could be included: 62 women and 22 men with a mean age of 74.7 years. Mean ASES score significantly increased from 47 preoperatively to 85.8 at the last follow-up (p = 0.001). The postoperative SST reached an average of 65.3 and the mean SSV was 83%. None of the variable parameters analyzed could be identified as a risk factor for a lower outcome defined as a SSV < 70. Three patients (3.6%) had a complication: one incomplete lower plexus lesion, one dislocation and one major hematoma. Surgical revision was needed in two cases (2.4%). Conclusion: The midterm clinical outcome of primary reverse total shoulder arthroplasty (rTSA) with an uncemented humeral short stem and a humeral inclination angle of 145° showed good-to-excellent results with a low complication and revision rate independent from a wide range of pre- and postoperative variables. PROMs are comparable to those reported for anatomic TSA with a low complication rate, different to historical studies especially with the Grammont design. Level of evidence: Treatment study, Level IV. [ABSTRACT FROM AUTHOR]

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Background: The Samilson-Prieto classification (SPC) depending on the humeral osteophyte length on a-pX-rays today is widely used to classify glenohumeral osteoarthritis in general. For treatment planning and prognosis, the patho-morphology of the glenoid and static posterior subluxation of the humeral head classified according to Walch is of much higher importance. Here, usually a CT or MRI scan is required for a correct classification. A possible correlation between both classifications is poorly explored. Without it, the complexity of the case might be mis-interpreted using the SPC. The aim of this study was to investigate such a correlation, i.e. whether it correlates with the glenoid deformity and degree of humeral head subluxation. Patients and methods: Radiological datasets (X-ray and CT or MRI) of 352 patients with primary OA of the shoulder were evaluated by two observers experienced in shoulder surgery. For the Samilson-Prieto classification, true a-p shoulder radiographs and for the modified Walch classification CT or MRI scans in the axial plane were interpreted using a validated method. To investigate a correlation between both classifications, the Fisher's exact test was used. For the interobserver reliability, the weighted kappa coefficient κ was determined. Results: For the Walch classification, both observers found a similar percentage for the different types, with decreasing numbers from normal (type A1) to severely altered glenoids In the Samilson-Prieto classification, OA grade I was predominant, while grade II and III showed a relatively equal distribution. Interobserver reliability was high both for the Walch classification with a κ 0.923 (95% confidence interval 0.892; 0.954) and) for the SPC with a κ 0.88 (95% confidence interval 0.843; 0.916). A correlation between the two classifications in Fischer's exact test could not be shown (p = 0.584). Discussion: Since there is no correlation between both, using the Samilson-Prieto classification alone might miss relevant prognostic factors in gleno-humeral OA. Adequate imaging of the glenoid morphology also in the axial plane is absolutely mandatory to understand the complexity and chose the right treatment for each patient. Level of evidence: Study of Diagnostic Test—Level II. [ABSTRACT FROM AUTHOR]

Loss-of-function mutations of SLC34A3 represent an established cause of a distinct renal phosphate wasting disorder termed hereditary hypophosphatemic rickets with hypercalciuria (HHRH). SLC34A3 encodes the renal phosphate transporter NaPi2c expressed at the apical brush border of proximal renal tubules. Substitution of p.Ser192Leu is one of the most frequent genetic changes among HHRH patients in Europe, but has never been systematically evaluated, clinically or on a cellular level. Identification of a 32-year-old female with a homozgyous c.575C>T, p.Ser192Leu substitution enabled a more comprehensive assessment of the impact of this missense variant. Clinically, the patient showed renal phosphate wasting and nephrocalcinosis without any bone abnormalities. Heterozygous carriers of deleterious SLC34A3 variants were previously described to harbor an increased risk of kidney stone formation and renal calcification. We hence examined the frequency of p.Ser192Leu variants in our adult kidney stone cohort and compared the results to clinical findings of previously published cases of both mono- and biallelic p.Ser192Leu changes. On a cellular level, p.Ser192Leu-mutated transporters localize to the plasma membrane in different cellular systems, but lead to significantly reduced transport activity of inorganic phosphate upon overexpression in Xenopus oocytes. Despite the reduced function in ectopic cellular systems, the clinical consequences of p.Ser192Leu may appear relatively mild, at least in our index patient, and can potentially be missed in clinical practice. [ABSTRACT FROM AUTHOR]

Nephrolithiasis (NL) affects 1 in 11 individuals worldwide and causes significant patient morbidity. We previously demonstrated a genetic cause of NL can be identified in 11-29% of pre-dominantly American and European stone formers. Pakistan, which resides within the Afro-Asian stone belt, has a high prevalence of nephrolithiasis (12%) as well as high rate of consanguinity (> 50%). We recruited 235 Pakistani subjects hospitalized for nephrolithiasis from five tertiary hospitals in the Punjab province of Pakistan. Subjects were surveyed for age of onset, NL recurrence, and family history. We conducted high-throughput exon sequencing of 30 NL disease genes and variant analysis to identify monogenic causative mutations in each subject. We detected likely causative mutations in 4 of 30 disease genes, yielding a likely molecular diagnosis in 7% (17 of 235) of NL families. Only 1 of 17 causative mutations was identified in an autosomal recessive disease gene. 10 of the 12 detected mutations were novel mutations (83%). SLC34A1 was most frequently mutated (12 of 17 solved families). We observed a higher frequency of causative mutations in subjects with a positive NL family history (13/109, 12%) versus those with a negative family history (4/120, 3%). Five missense SLC34A1 variants identified through genetic analysis demonstrated defective phosphate transport. We examined the monogenic causes of NL in a novel geographic cohort and most frequently identified dominant mutations in the sodium-phosphate transporter SLC34A1 with functional validation. [ABSTRACT FROM AUTHOR]

Teleost fish are excellent models to study the phylogeny of the slc34 gene family, Slc34-mediated phosphate (Pi) transport and how Slc34 transporters contribute Pi homeostasis. Fish need to accumulate Pi from the diet to sustain growth. Much alike in mammals, intestinal uptake in fish is partly a paracellular and partly a Slc34-mediated transcellular process. Acute regulation of Pi balance is achieved in the kidney via a combination of Slc34-mediated secretion and/or reabsorption. A great plasticity is observed in how various species perform and combine the different processes of secretion and reabsorption. A reason for this diversity is found in one or two whole genome duplication events followed by potential gene loss; consequently, teleosts exhibit distinctly different repertoires of Slc34 transporters. Moreover, due to habitats with vastly different salinity, teleosts face the challenge of either preserving water in a hyperosmotic environment (seawater) or excreting water in hypoosmotic freshwater. An additional challenge in understanding teleost Pi homeostasis are the genome duplication and retention events that diversified peptide hormones such as parathyroid hormone and stanniocalcin. Dietary Pi and non-coding RNAs also regulate the expression of piscine Slc34 transporters. The adaptive responses of teleost Slc34 transporters to e.g. Pi diets and vitamin D are informative in the context of comparative physiology, but also relevant in applied physiology and aquaculture. In fact, Pi is essential for teleost fish growth but it also exerts significant adverse consequences if over-supplied. Thus, investigating Slc34 transporters helps tuning the physiology of commercially valuable teleost fish in a confined environment. [ABSTRACT FROM AUTHOR]

In the pre-genomic era, the cloning of a cDNA represented a significant achievement, particularly if the gene of interest encoded a membrane protein. At the time, molecular probes such as partial peptide sequences, suitable nucleic acid sequences, or antibodies were unavailable for most proteins and the "sodium-phosphate transporter" was no exception. In contrast, brush-border membrane vesicles and epithelial cell culture experiments had established a reliable set of functional hallmarks that described Na-dependent phosphate transport activity in some detail. Moreover, aspects of hormonal regulation of phosphate homeostasis could be recapitulated in these model systems. Expression cloning elegantly combined functional protein expression in Xenopus laevis oocytes with molecular biology to overcome the lack of molecular probes. [ABSTRACT FROM AUTHOR]

Thermochemical energy storage uses reversible thermochemical reactions to store and release heat, representing a promising technology for energy conservation and utilizing fluctuating renewable energy sources and waste heat. Many recent studies have focused on determination of the enthalpy of reaction of possible thermochemical materials (TCM) based on thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). So far, comparatively few attempts have been made to characterize the apparent specific heat capacity at constant pressure cpappT of the investigated TCM. The purpose of this study is to outline a measurement and analysis procedure to evaluate cpappT of powdery TCM. The procedure is presented focusing on two metal hydroxides Ca(OH)2 and Mg(OH)2. Preliminary TGA experiments were conducted to identify reaction-free temperature intervals and mass change. Starting from the metal hydroxide, subsequent DSC experiments with two consecutive heating and cooling cycles were carried out to determine cpappT of the initial hydroxide and the oxide product. Three separate DSC runs for each candidate enable an evaluation of measurement uncertainty, and cpappT results were compared to available literature data. Preliminary TGA experiments have shown that the applied heating rate β has a strong effect on the measured dehydration reaction. This result influences the consecutive cpappT interpretation of the metal hydroxides. Analysis of the measured cpappT data compared to literature show good agreement for both metal hydroxides and oxides. Overlapping endotherm effects, which are not part of cpT, have to be considered for further thermal conductivity calculations. [ABSTRACT FROM AUTHOR]

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Introduction: Prognosis of Her2-positive breast cancer has changed since the introduction of trastuzumab for treatment in metastatic and early breast cancer. It was described to be even better compared to prognosis of Her2-negative metastatic breast cancer. The purpose of this study was to evaluate the effect of trastuzumab in our cohort. Besides the effect of adjuvant pretreatment with trastuzumab on survival of patients with metastatic Her2-positive breast cancer was analyzed.Methods: All patients with primary breast cancer of the Regional Breast Cancer Center Dresden diagnosed during the years 2001-2013 were analyzed for treatment with or without trastuzumab in the adjuvant and in the metastatic treatment setting using Kaplan-Meier survival estimation and Cox regression. Age and tumor stage at time of first diagnosis of breast cancer as well as hormone receptor status, grading, time, and site of metastasis at first diagnosis of distant metastatic disease were analyzed.Results: Of 4.481 female patients with primary breast cancer, 643 presented with metastatic disease. Her2-positive status was documented in 465 patients, including 116 patients with primary or secondary metastases. Median survival of patients with Her2-positive primary metastatic disease was 3.0 years (95% CI 2.3-4.0). After adjustment for other factors, survival was better in patients with Her2-positive breast cancer with trastuzumab therapy compared to Her2-negative metastatic disease (HR 2.10; 95% CI 1.58-2.79). Analysis of influence of adjuvant therapy with and without trastuzumab by Kaplan-Meier showed a trend for better survival in not pretreated patients. Median survival was highest in hormone receptor-positive Her2-positive (triple-positive) primary metastatic breast cancer patients with 3.3 years (95% CI 2.3-4.6).Conclusion: Prognosis of patients with Her2-positive metastatic breast cancer after trastuzumab treatment is more favorable than for Her2-negative breast cancer. The role of adjuvant chemotherapy with or without trastuzumab warrants further research. Survival is best in triple-positive metastatic breast cancer. This will effect counseling at the time of first diagnosis of metastatic breast cancer. [ABSTRACT FROM AUTHOR]

Clinical outcome of patients with stage IV breast cancer is dependent on tumor biology, extent, and localization of metastases. Routine imaging diagnostics for distant metastasis is not recommended by the national guidelines for breast cancer follow-up. In this study, we evaluated different patterns of metastases of cancer subtypes in order to generate hypotheses on individualization of follow-up after breast cancer in the adjuvant setting. Patients of the Regional Breast Cancer Center Dresden diagnosed within the years 2006-2011 were classified into the five intrinsic subtypes luminal A (ER+, Her2−, G1/2), luminal B/Her2 negative (ER+, Her2−, G3), triple positive (ER+, PR+, Her2+), Her2-enriched (ER−, Her2+), and triple negative (ER−, PR−, Her2−) and with a median follow-up of 45 months. Tumor stage at time of first diagnosis of breast cancer as well as time and site of metastasis at first diagnosis of distant metastatic disease was analyzed. Tumor specimen of 2284 female patients with primary breast cancer was classified into five subtypes. Distant recurrence-free survival at 3 years was most unfavorable in Her2-enriched (66.8 %), triple negative (75.9 %), and triple-positive breast cancer (81.7 %). The same subtypes most frequently presented with visceral metastases only at first presentation: Her2-enriched 46.9 %, triple negative 45.5 %, and triple-positive breast cancer 37.5 %. Longest median survival of 2.3 years was seen in luminal A and in Her2-enriched metastatic disease, respectively. Median survival was significantly better in the luminal A, Her2-enriched, and triple-positive subtype compared to triple-negative breast cancer ( p < 0.005). Differences in time to metastatic disease, first localization of metastases, and overall survival after diagnosis of metastatic disease were shown. Considering new targeted therapies and the option of surgery of oligometastases, screening for visceral metastases might be reasonable after diagnosis of Her2-positive subtypes. [ABSTRACT FROM AUTHOR]

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Heat shock and other environmental stresses rapidly induce transcriptional responses subject to regulation by a variety of post-translational modifications. Among these, poly(ADP-ribosyl)ation and sumoylation have received growing attention. Here we show that the SUMO E3 ligase PIASy interacts with the poly(ADP-ribose) polymerase PARP-1, and that PIASy mediates heat shock-induced poly-sumoylation of PARP-1. Furthermore, PIASy, and hence sumoylation, appears indispensable for full activation of the inducible HSP70.1 gene. Chromatin immunoprecipitation experiments show that PIASy, SUMO and the SUMO-conjugating enzyme Ubc9 are rapidly recruited to the HSP70.1 promoter upon heat shock, and that they are subsequently released with kinetics similar to PARP-1. Finally, we provide evidence that the SUMO-targeted ubiquitin ligase RNF4 mediates heat-shock-inducible ubiquitination of PARP-1, regulates the stability of PARP-1, and, like PIASy, is a positive regulator of HSP70.1 gene activity. These results, thus, point to a novel mechanism for regulating PARP-1 transcription function, and suggest crosstalk between sumoylation and RNF4-mediated ubiquitination in regulating gene expression in response to heat shock. [ABSTRACT FROM AUTHOR]

The aim of this study was to elucidate mechanisms of Pi handling in toads ( Bufo bufo). We introduced toads to experimental solutions of various [Pi] and high Pi diets and measured urine and lymph [Pi]. Both lymph and urine [Pi] increased with increasing Pi loads, indicating Pi absorption across skin and intestine. An initial fragment of a NaPi-II type transporter was amplified from kidney, and the full-length sequence was obtained. The protein showed the molecular hallmarks of NaPi-IIb transporters. When expressed in Xenopus oocytes the clone showed unusual pH dependence, but apparent affinity constants for Pi and Na+ were in the range of other NaPi-II transporters. Expression profiling showed that the transporter was present in skin, intestine and kidney. Reverse transcription–polymerase chain reaction assays on dissected renal tubules indicated expression in the collecting duct system. Collecting tubules and ducts were isolated, perfused and microelectrode recordings showed electrogenic Pi transport in apical and basolateral membranes. Taken together, our results show that Pi is handled by intestine, kidney and skin. The presently cloned NaPi-IIb is a likely candidate involved in Pi absorption across these epithelia. In addition, electrophysiological experiments suggest that the collecting duct system plays an important role in Pi homeostasis. [ABSTRACT FROM AUTHOR]

Even slight disruptions in the balance of serum Pi can lead to major disorders ranging from cardiovascular and kidney disease if serum phosphate is too high to bone demineralisation if serum Pi is too low [[5]]. Pi reabsorption in the distal parts of the renal tubule, "fine-tuning Pi excretion in the urine" has long been established, though the molecular mediators of distal Pi uptake have not been identified [[1]]. Motta and Silva et al. also demonstrate that mice fed a high Pi diet did not appear to have altered Pi transporter mRNA levels compared with those fed a low Pi diet. [Extracted from the article]

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The epithelial Na/Pi cotransporter (NaPi-II) is instrumental in maintaining phosphate (Pi) homeostasis in vertebrates. Hormones and metabolic factors (PTH, Pi availability) that acutely influence renal Pi excretion have been demonstrated to target NaPi-II expression. Upon stimulation, newly synthesized transporter molecules become integrated into the brush-border membrane to increase the V max of Pi uptake; reduction of Pi reabsorption is achieved by endocytosis of NaPi-II followed by lysosomal degradation of the protein. The long-term regulation of the protein is less well studied. Only recently, regulatory elements for vitamin D3 and Pi have been identified in the promoter region of the npt gene. However, signaling pathways leading to the activation of these regulatory sequences need to be established. Other reports suggested messenger RNA stability to play a role in the medium range regulation of NaPi-II expression. Recent findings in our laboratory added to the complex picture of npt gene regulation. We have identified npt-related endogenous antisense transcripts from mouse, zebrafish, and winter flounder. The two fish transcripts have been cloned and characterized; the mouse homolog has only very recently been detected. The transcripts are devoid of an open reading frame and appear in different splice forms. The evolutionary conservation of bidirectional transcription of the npt gene implies a regulatory function for the antisense transcript. In order to test the functional consequences of bidirectional transcription, we coexpressed sense and the antisense transcripts from zebrafish in Xenopus oocytes. Pi transport activity was reduced as a result of the presence of antisense RNA. Re-extraction of the RNA from injected oocytes followed by Northern blot revealed that the coexpression had no significant effect on the stability of either transcript. We concluded that the antisense mRNA interfered with the translation of the transporter if coexpressed in the Xenopus system. However, the regulatory mechanism(s) involving the npt-related antisense transcript is expected to be much more complicated in vivo, (i.e., requiring supplementary factors like double-stranded RNA recognizing proteins or specific RNases). It is planned to test this hypothesis by a transgenic zebrafish approach and/or knockout mice. [ABSTRACT FROM AUTHOR]

On inflamed endothelium the cell surface protein E-selectin isexpressed which supports the initial process of attachment –capturing and rolling of leukocytes. A recombinant CHO cell linesecreting a soluble E-selectin-IgG chimera was cultivated competitively under serum free conditions in three different bioreactor systems: a 1 l Super-Spinner, a 2 l stirred tank bioreactor equipped with a spinfilter, and a 100 l stirred tankbioreactor. In the smallest system 25.4 mg E-selectin-IgG wereproduced in 62 days using a repeated batch process whileachieving a maximal viable cell density of 3.7 × 106 cells ml-1. Using continuous perfusion mode a total amount of35.2 mg were produced with a maximal viable cell density of1.65 × 107 cells ml-1 in the 2 l bioreactor within 29 days. Large scale cultivation in a 100 l stirred tankbioreactor yielded 105.6 mg in three batches with a maximal viable cell density of 9.7 × 105 cells ml-1 within 15 days. After removal of the cells by continuous centrifugation and a depth filter clearance step, the supernatants were concentrated via ultra filtration. Purificationwas performed by affinity chromatography with rProtein A. Integrity of the E-selectin-IgG protein was checked with SDS PAGE. Its activity was verified in a cellular adhesion assay performed with HL-60 cells and a recombinant CHO cell line expressing membrane-anchored E-selectin constitutively, and E-selectin expressing HUVECs, respectively. Soluble E-selectin-IgG was used to block adhesion to these cell layerscompetitively. A concentation of 18.8 and 37.5 μg ml-1was sufficient to reduce the amount of adhering HL-60 cells to 50% on CHO and HUVEC layers, respectively. [ABSTRACT FROM AUTHOR]

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Recently, we have isolated a complementary DNA most likely related to rabbit kidney cortex brush border membrane sodium/phosphate (Na/Pi) cotransport activity [NaPi-1 (1)]. To further elucidate the cellular mechanisms involved in dietary 'adaptation' of renal Na/Pi contransport, we have exposed young rabbits for 2 weeks to either a low phosphate (Pi) diet (LPD) or a high Pi diet (HPD). Initial linear uptake of Na/Pi cotransport in isolated brush border membrane vesicles was increased in rabbits on a LPD compared with those on a HPD. Injection of equal amounts of total mRNA isolated from kidney cortex of LPD or HPD rabbits into Xenopus laevis oocytes resulted in a higher stimulation of Na-dependent oocyte Pi uptake in LPD than HPD preparations. No difference in the content of 'specific' mRNA (NaPi-1 cDNA probe, Northern blots) and of the content of the 'specific' brush border membrane protein (NaPi-1 antipeptide antibody, Western blots) between LPD and HPD preparations was observed. We conclude that 'chronic' dietary Pi deprivation leads to a protein synthesis-dependent alteration of Na/Pi cotransport activity which does not involve a change in the total amount of a protein related to the recently cloned NaPi-1 protein. [ABSTRACT FROM AUTHOR]

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Copyright of Chemical Monthly / Monatshefte für Chemie is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The complexation of C- or C-symmetrical torsional isomeric benzene derivatives may result in residual stereoisomeric complexes if the ligand is converted to its enantiomer by the torsional motion and if the complexation site is not within the symmetry axis (C axis). The resulting residual stereoisomers are well suited for optical resolution, especially if the equilibrium is strongly in favor of one isomer. [ABSTRACT FROM AUTHOR]

Expression of Na-dependent transport of phosphate (P) was analysed in Xenopus laevis oocytes after injection of poly(A)-rich RNA isolated from the duodenal mucosa of rabbits with increased levels of 1,25-(OH)-VitD (injection of vitamin D or low-P diet) or from control animals. In parallel, the effect of elevated levels of vitamin D was studied in isolated duodenal brush-border membrane vesicles. In brush-border membrane vesicles, the rate of Na-P cotransport was found to be doubled after 1,25-(OH)-VitD injections while Na - d-glucose cotransport (measured as a control) was not altered. In X. laevis oocytes, Na-dependent P uptake was increased after injection of poly(A)-rich RNA isolated from duodenal mucosa of animals with increased levels of 1,25-(OH)-VitD but not after injection of poly(A)-rich RNA isolated from control animals; between the two groups of mRNA no difference in the expression of the Na - d-glucose transport system was observed. Sucrose density gradient fractionation suggests that mRNA species related to the increased Na-dependent P uptake are of average chain lengths between 2×10 and 3×10 bases (2-3 kb). It is concluded that in duodenal enterocytes 1,25-(OH)-VitD increases the content of mRNA species of 2-3 kb that might be involved either directly in Na-P cotransport or at least in controlling its activity. [ABSTRACT FROM AUTHOR]

Narrowing is a complete unification procedure for equational theories defined by canonical term rewriting systems. It is also the operational semantics of various logic and functional programming languages. In Ref. 3), we introduced the LSE narrowing strategy, which is complete for arbitrary canonical rewriting systems and optimal in the sense that two different LSE narrowing derivations cannot generate the same narrowing substitution. LSE narrowing improves all previously known strategies for the class of arbitrary canonical systems. LSE narrowing detects redundant derivations by reducibility tests. According to their definition, LSE narrowing steps seem to be very expensive, because a large number of subterms has to be tested. In this paper, we show that many of these subterms are identical. We describe how left-to-right basic occurrences can be used to identify and exclude these identical subterms. This way, we can drastically reduce the number of subterms that have to be tested. Based on these theoretical results, we develop an efficient implementation of LSE narrowing. [ABSTRACT FROM AUTHOR]

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Xenopus laevis oocytes have been used for the expression of a renal, pyrimidine-selective, Na-nucleoside cotransporter (N2). As compared to its uptake in water-injected oocytes, Na-dependent thymidine uptake was enhanced in a time- and dose- dependent manner in oocytes injected with rat renal cortex total poly(A) RNA. An increased uptake was also observed after injection of size fractionated rat renal cortex poly(A) RNA (2-3 kb). Consistent with the selectivity of the N2 nucleoside transporter, cytidine significantly inhibited Na-dependent thymidine uptake in oocytes injected with total poly(A) RNA whereas guanosine and formycin B did not. Na-dependent thymidine uptake was also enhanced in oocytes injected with size fractionated human renal cortex poly(A) RNA (2-3 kb). The above data demonstrate functional expression of renal cortex, Na-nucleoside cotransporters in Xenopus laevis oocytes. [ABSTRACT FROM AUTHOR]