Abstract INTRODUCTION: The metastasis of a prostate tumor is the leading cause of prostate cancer deaths. Therefore the ability to prevent or limit the malignant potential of tumor cells remains a significant and clinically relevant goal. Prostate-Derived ETS Factor (PDEF) is a transcription factor which regulates multiple gene products involved in prostate tumorigenesis. There is an inverse correlation between the levels of PDEF and tumor aggressiveness. In addition, PDEF protein expression is inversely correlated with increasing tumor aggressiveness (increasing Gleason Score), suggesting that PDEF may be a tumor metastasis suppressor. Since PC3 cells express very low levels of PDEF and metastasize well in metastasis models, they serve as a model to determine: 1) if PDEF over-expression affects the metastatic capacity of these cells in mouse models of metastasis and 2) which PDEF-regulated gene product(s) may contribute to metastatic colonization. METHODS: Standard migration (scratch), invasion (Boyden chamber), clonogenic, and RNA to cDNA RT-PCR assays were performed. Cells were stably transfected to express Luciferase. In vivo, 1 x 106 cells were injected in the subcutaneous, tail vein, and intra-cardic mouse models and at various time points there after, cells are non-invasively monitored by IVIS over a twelve week period. RESULTS: Utilizing in vitro migration, invasion, and clonogenic assays we demonstrated that PDEF over-expression resulted in decreased tumor aggressiveness. PDEF re-expression differentially regulated MMP9 and FAK compared to parental cells. Surprisingly, PDEF expression had no effect on in vivo subcutaneous xenograft tumor growth; however, using both tail-vein and intra-cardiac injection mouse metastasis models we demonstrate that PC3 cells as well as PDEF over-expressing PC3 cells disperse and home in around abdominal cavity. We observed that over time PC3 cells survived and reestablished new colonies. Conversely, PDEF over-expression significantly and dramatically decreased the ability of PC3 cells to colonize metastatic sites compared to parental cells. Twelve weeks post tumor cell injection, mice injected with PC3 cells had well established micro-metastasis, while mice injected with PDEF-over-expressing PC3 cells had no detectible metastasis. CONCLUSIONS: These studies demonstrate for the first time that the re-expression of PDEF decreases the metastatic capacity of prostate cancer cells in vivo, while having no effect on subcutaneous tumor cell growth, thus establishing PDEF as a tumor metastasis suppressor gene. Therefore, future studies to determine how PDEF is silenced in advance disease may lead to increased mechanistic insight into the metastatic process and eventually to several therapeutic options for the re-expression of PDEF or the inhibition of PDEF-regulated gene products to decrease or perhaps prevent prostate tumor metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3431. doi:1538-7445.AM2012-3431

Vascular calcification is a common complication in atherosclerosis. Bone morphogenetic protein-2 (BMP-2) plays an important role in atherosclerotic vascular calcification. The aim of this study was to determine the effect of oxidized low density lipoprotein (oxLDL) on BMP-2 protein expression in human coronary artery endothelial cells (CAECs), the roles of Toll-like receptor (TLR) 2 and TLR4 in oxLDL-induced BMP-2 expression, and the signaling pathways involved. Human CAECs were stimulated with oxLDL. The roles of TLR2 and TLR4 in oxLDL-induced BMP-2 expression were determined by pretreatment with neutralizing antibody, siRNA, and overexpression. Stimulation with oxLDL increased cellular BMP-2 protein levels in a dose-dependent manner (40–160 μg/ml). Pretreatment with neutralizing antibodies against TLR2 and TLR4 or silencing of these two receptors reduced oxLDL-induced BMP-2 expression. Overexpression of TLR2 and TLR4 enhanced the cellular BMP-2 response to oxLDL. Furthermore, oxLDL was co-localized with TLR2 and TLR4. BMP-2 expression was associated with activation of nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (MAPK), and extracellular signal-regulated kinase (ERK)1/2. Inhibition of NF-κB and ERK1/2 reduced BMP-2 expression whereas inhibition of p38 MAPK had no effect. In conclusion, oxLDL induces BMP-2 expression through TLR2 and TLR4 in human CAECs. The NF-κB and ERK1/2 pathways are involved in the signaling mechanism. These findings underscore an important role for TLR2 and TLR4 in mediating the BMP-2 response to oxLDL in human CAECs and indicate that these two immunoreceptors contribute to the mechanisms underlying atherosclerotic vascular calcification.

BACKGROUND:Prostate-derived Ets factor (PDEF) is expressed in tissues of high epithelial content including prostate, although its precise function has not been fully established. Conventional therapies produce a high rate of cure for patients with localized prostate cancer, but there is, at present, no effective treatment for intervention in metastatic prostate cancer. These facts underline the need to develop new approaches for early diagnosis of aggressive prostate cancer patients, and mechanism based anti-metastasis therapies that will improve the outlook for hormone-refractory prostate cancer. In this study we evaluated role of prostate-derived Ets factor (PDEF) in prostate cancer.RESULTS:We observed decreased PDEF expression in prostate cancer cell lines correlated with increased aggressive phenotype, and complete loss of PDEF protein in metastatic prostate cancer cell lines. Loss of PDEF expression was confirmed in high Gleason Grade prostate cancer samples by immuno-histochemical methods. Reintroduction of PDEF profoundly affected cell behavior leading to less invasive phenotypes in three dimensional cultures. In addition, PDEF expressing cells had altered cell morphology, decreased FAK phosphorylation and decreased colony formation, cell migration, and cellular invasiveness. In contrast PDEF knockdown resulted in increased migration and invasion as well as clonogenic activity. Our results also demonstrated that PDEF downregulated MMP9 promoter activity, suppressed MMP9 mRNA expression, and resulted in loss of MMP9 activity in prostate cancer cells. These results suggested that loss of PDEF might be associated with increased MMP9 expression and activity in aggressive prostate cancer. To confirm results we investigated MMP9 expression in clinical samples of prostate cancer. Results of these studies show increased MMP9 expression correlated with advanced Gleason grade. Taken together our results demonstrate decreased PDEF expression and increased MMP9 expression during the transition to aggressive prostate cancer.CONCLUSIONS:These studies demonstrate for the first time negative regulation of MMP9 expression by PDEF, and that PDEF expression was lost in aggressive prostate cancer and was inversely associated with MMP9 expression in clinical samples of prostate cancer. Based on these exciting results, we propose that loss of PDEF along with increased MMP9 expression should serve as novel markers for early detection of aggressive prostate cancer.

You have accessJournal of Urology1 Apr 2009RECIPROCAL REGULATION OF FOCAL ADHESION KINASE (FAK) AND EXTRACELLULAR SIGNAL REGULATED KINASE (ERK) IN PROSTATE CANCER CLONOGENIC ACTIVITY AND INVASION Paul D Maroni, Thomas R Johnson, Lakshmipathi Khandrika, Binod Kumar, Sweaty Koul, Randall B Meacham, and Hari K Koul Paul D MaroniPaul D Maroni More articles by this author , Thomas R JohnsonThomas R Johnson More articles by this author , Lakshmipathi KhandrikaLakshmipathi Khandrika More articles by this author , Binod KumarBinod Kumar More articles by this author , Sweaty KoulSweaty Koul More articles by this author , Randall B MeachamRandall B Meacham More articles by this author , and Hari K KoulHari K Koul More articles by this author View All Author Informationhttps://doi.org/10.1016/S0022-5347(09)60749-7AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "RECIPROCAL REGULATION OF FOCAL ADHESION KINASE (FAK) AND EXTRACELLULAR SIGNAL REGULATED KINASE (ERK) IN PROSTATE CANCER CLONOGENIC ACTIVITY AND INVASION." The Journal of Urology, 181(4S), p. 263 © 2009 by American Urological AssociationFiguresReferencesRelatedDetails Volume 181Issue 4SApril 2009Page: 263 Advertisement Copyright & Permissions© 2009 by American Urological AssociationMetricsAuthor Information Paul D Maroni More articles by this author Thomas R Johnson More articles by this author Lakshmipathi Khandrika More articles by this author Binod Kumar More articles by this author Sweaty Koul More articles by this author Randall B Meacham More articles by this author Hari K Koul More articles by this author Expand All Advertisement PDF downloadLoading ...

Overexpression of focal adhesion kinase (FAK) has been well correlated with tumor development and/or the maintenance of tumor phenotype. In addition, inappropriate activation of the extracellular regulated kinase (ERK) signaling pathway is common to many human cancers. In the present study, we investigated the interplay between FAK and ERK in androgen-independent prostate cancer cells (PC3 and DU145 cells). We observed that suppression of FAK expression using small interfering RNA–mediated knockdown decreased the clonogenic activity, whereas overexpression of FAK increased it. We also observed that detachment of PC3 and DU145 cells from their substrate induced tyrosine phosphorylation of FAK. ERK knockdown diminished FAK protein levels and tyrosine phosphorylation of FAK as well as FAK promoter-reporter activity. We also tested the effect of MEK inhibitors and small interfering RNA–mediated knockdown of ERK1 and/or ERK2 on cell proliferation, invasiveness, and growth in soft agar of PC3 and DU145 cells. Inhibition of ERK signaling grossly impaired clonogenicity as well as invasion through Matrigel. However, inhibition of ERK signaling resulted in only a modest inhibition of 3H-thymidine incorporation and no effect on overall viability of the cells or increased sensitivity to anoikis. Taken together, these data show, for the first time, a requirement for FAK in aggressive phenotype of prostate cancer cells; reveal interdependence of FAK and ERK1/2 for clonogenic and invasive activity of androgen-independent prostate cancer cells; suggest a role for ERK regulation of FAK in substrate-dependent survival; and show for the first time, in any cell type, the regulation of FAK expression by ERK signaling pathway. (Mol Cancer Res 2008;6(10):1639–48)

Previous studies demonstrated that osteogenic mediators, in particular bone morphogenetic protein-2 (BMP-2), play an important role in the progression of atherosclerosis and vascular calcification. Although it is known that the atherogenic factor oxidized low density lipoprotein (oxLDL) up-regulates BMP-2 expression in endothelial cells, the signaling mechanisms involved are not well understood. In macrophages, oxLDL up-regulates Toll-like receptor 4 (TLR4) expression, and this innate immunoreceptor appears to be involved in oxLDL-induced macrophage actin polymerization. We earlier found that stimulation of TLR4 with lipopolysac-charide (LPS) increases cellular BMP-2 protein levels in human aortic valve interstitial cells. We hypothesized that the TLR4 pathway plays a central role in mediating BMP-2 expression in human coronary artery endothelial cells (CAECs). The purposes of this study were to examine whether TLR4 mediates oxLDL- and/or LPS-induced BMP-2 expression in human CAECs, and to determine whether the p38 and/or p44/42 MAPKs are involved. Methods and results : Stimulating human CAECs with LPS (E. coli 0111:B4, 200 ng/ml) for 24 h up-regulated BMP-2 protein expression. Similarly, stimulation with oxLDL (from human plasma, CuSO 4 -oxidized, LPS-free, 40–160 μg/ml) for 24 h induced BMP-2 expression in a dose-dependent manner. Pretreatment with either TLR4-neutralizing antibody or TLR4 siRNA significantly attenuated oxLDL-induced BMP-2 expression and abrogated the effect of LPS on BMP-2 expression. Over-expression of TLR4 enhanced the cellular BMP-2 response to oxLDL and LPS. Further, immunofluorescent staining co-localized oxLDL with TLR4. Although LPS induced ICAM-1 expression in human CAECs, oxLDL had no effect, indicating that oxLDL and LPS have different pro-inflammatory effects. BMP-2 expression was associated with activation of p38 and p44/42 MAPKs. Inhibiting p44/42, but not p38, reduced BMP-2 expression. Conclusions: In human CAECs, TLR4 plays a central role in regulating BMP-2 expression induced by either oxLDL or LPS, and the signaling mechanism involves the p44/42 MAPK pathway. These novel findings underscore an important role of TLR4 in atherosclerosis and vascular calcification.

You have accessJournal of UrologyPodium, Tuesday, May 23, 2006, 8:00 - 10:00 am1 Apr 20061043: A Rational Gene Therapy for Primary Hyperoxaluria-1 (PH-1): Cloning, Cellular Transfection and Sub Cellular Localization of AGT-GFP-Fusion Protein Sweaty Koul, Thomas Johnson, Randall B. Meacham, Paramjit S. Chandhoke, and Hari K. Koul Sweaty KoulSweaty Koul More articles by this author , Thomas JohnsonThomas Johnson More articles by this author , Randall B. MeachamRandall B. Meacham More articles by this author , Paramjit S. ChandhokeParamjit S. Chandhoke More articles by this author , and Hari K. KoulHari K. Koul More articles by this author View All Author Informationhttps://doi.org/10.1016/S0022-5347(18)33268-3AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "1043: A Rational Gene Therapy for Primary Hyperoxaluria-1 (PH-1): Cloning, Cellular Transfection and Sub Cellular Localization of AGT-GFP-Fusion Protein." The Journal of Urology, 175(4S), pp. 335–336 © 2016 by American Urological AssociationFiguresReferencesRelatedDetails Volume 175Issue 4SApril 2006Page: 335-336 Advertisement Copyright & Permissions© 2016 by American Urological AssociationMetricsAuthor Information Sweaty Koul More articles by this author Thomas Johnson More articles by this author Randall B. Meacham More articles by this author Paramjit S. Chandhoke More articles by this author Hari K. Koul More articles by this author Expand All Advertisement PDF DownloadLoading ...

The Losee repair controls rotational subluxation of the lateral femoral condyle, or pivot shift, but does not reliably eliminate Lachman laxity. Despite this surgical limitation, many patients who were operated on continued to do high-demand activities at the last followup. We hypothesized that Lachman findings alone did not predict poor surgical outcome or progression to osteoarthritis. We report on 87 patients evaluated at an average of 9 years (range, 5-21 years) postoperatively. Prospectively collected examinations and radiographic, subjective, and objective outcome measures were recorded and statistically evaluated. The presence of a postoperative pivot shift or residual varus laxity correlated with poor patient subjective evaluations and poor scoring outcomes. Lachman laxity with an absent pivot shift had no correlation with the outcome measures or onset of radiographic progression to osteoarthritis. Meniscectomy, additional knee surgery, increased valgus or varus laxity, and time from injury until the final radiograph positively correlated with the onset of osteoarthritis. Elimination of the pivot shift was necessary to achieve successful relief of symptoms and functional outcome. In the absence of a pivot shift, Lachman laxity was not solely predictive of poor outcomes. Level of Evidence: Prognostic study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence.

Background: Primary hyperoxaluria-type 1 (PH-1) is a rare autosomal recessive disorder of glyoxalate metabolism caused by deficiency in the liver-specific peroxisomal enzyme alanine-glyoxalate transaminase 1 (AGT) resulting in the increased oxidation of glyoxalate to oxalate. Accumulation of oxalate in the kidney and other soft tissues results in loss of renal function and significant morbidity. The present treatment options offer some relief in the short term, but they are not completely successful. In the present study, we tested the feasibility of corrective gene therapy for this metabolic disorder. Methods: A cDNA library was made from HepG2 cells. PCR primers were designed for the AGT sequence with modifications to preclude mistargeting during gene delivery. Amplified AGT cDNA was cloned as a fusion protein with green fluorescent protein (GFP) using the vector EGFP-C1 (Clontech) for monitoring subcellular distribution. Sequence and expression of the fusion protein was verified. Fusion protein vectors were transfected into hepatocytes by liposomal transfection. AGT expression and subcellular distribution was monitored by GFP fluorescence. Results: HepG2 cells express full-length mRNA coding for AGT as confirmed by insert size as well as sequence determination. Selective primers allowed us to generate a modified recombinant GFP-AGT fusion protein. Cellular transfections with Lipofectamine resulted in transfection efficiencies of 60–90%. The recombinant AGT did localize to peroxisomes as monitored by GFP fluorescence. Conclusions: The results demonstrate preliminary in vitro feasibility data for AGT transfection into the hepatocytes. To the best of our knowledge, this is the first study to attempt recombinant AGT gene therapy for treatment of primary hyperoxaluria-1.

Quantitative indicators of biological integrity are needed for streams in the Great Plains of North America, but it was not known whether the index of biotic integrity (IBI) approach would be effective in this semiarid region. Great Plains streams have a depauperate and tolerant ichthyofauna and highly variable physicochemical conditions that may mask the effects of non-point-source pollution and stream habitat degradation. We developed an IBI based on fish assemblages by screening metrics for range, responsiveness to human influence, precision, and lack of redundancy; we then tested the IBI's ability to detect anthropogenic effects by validating the index with an independent data set. The IBI was composed of 10 metrics based on species richness and composition, trophic and reproductive guilds, and age structure. These 10 metrics had many significant correlations with substrate and water chemistry variables but had fewer significant correlations with riparian condition and watershed variables. Of the watershed variables, road density had the highest number of significant correlations with final IBI metrics. The IBI was validated by demonstrating its responsiveness to aggregate measures of human influence, site-level habitat, and water chemistry, and its lack of responsiveness to factors that varied naturally, such as stream size and site elevation. The IBI was also temporally stable within and between years during repeat visits to a subset of sampled reaches. This IBI can be used as a measure of biological integrity for management of prairie streams faced with threats such as introduced species, intensive agriculture, grazing, and coalbed natural gas extraction. Although we developed this IBI based on data from Montana prairie streams only, our IBI can probably serve as a framework for other North American plains streams and our results suggest that the IBI approach may be useful in other semiarid regions of the world.

We demonstrate that PS-341, a small molecule inhibitor of the proteasome, markedly sensitizes resistant prostate, colon, and bladder cancer cells to TNF-like apoptosis-inducing ligand (TRAIL)-induced apoptosis irrespective of Bcl-xL overexpression. PS-341 treatment by itself does not affect the levels of Bax, Bak, caspases 3 and 8, c-Flip or FADD, but elevates levels of TRAIL receptors DR4 and DR5. This increase in receptor protein levels is associated with the ubiquitination of the DR5 protein. When PS-341 is combined with TRAIL, the levels of activated caspase 8 and cleaved Bid are substantially increased. In Bax-negative TRAIL-resistant HC-4 colon cancer cells, the combination of PS-341 and TRAIL overcomes the block to activation of the mitochondrial pathway and causes SMAC and cytochrome c release followed by apoptosis. Similarly, murine embryonic fibroblasts lacking Bax undergo apoptosis when exposed to the combination of PS-341 and TRAIL; however, fibroblasts lacking Bak are significantly resistant. Taken together, these findings indicate that PS-341 enhances TRAIL-induced apoptosis by increasing the cleavage of caspase 8, causing Bak-dependent release of mitochondrial proapoptotic proteins.

Treatment of leukemic cells with phorbol 12-myristate 13-acetate (PMA) induces a short-lived phosphorylation and activation of stress-activated protein kinase (SAPK) and cellular differentiation. To investigate whether the rapid deactivation of SAPK results from dephosphorylation by dual-specificity phosphatases (DSPs), we studied regulation of the DSP hVH5 and its murine orthologue M3/6 in K562 human leukemia cells. PMA treatment rapidly induced hVH5 transcripts in these cells, and induced expression of M3/6 completely inhibited PMA-stimulated phosphorylation of SAPK, suggesting a feedback loop to control SAPK activity. Using both stable cell lines and transient transfection we demonstrate that activation of SAPK rapidly stimulated phosphorylation of M3/6. This phosphorylation did not regulate the half-life of total cellular M3/6. hVH5 and M3/6 shares with all sequenced mammalian DSPs an amino acid motif,XILPXLXL, located approximately 80 amino acids from the active site. The hVH5-M3/6 sequence, RILPHLYL, shares significant homology with the SAPK binding site of the c-Jun protein, called the delta domain. This motif was found to be important for DSP function, because deletion of RILPHLYL inhibits SAPK-mediated phosphorylation of M3/6, and deletion of this sequence or mutation of the LYL portion blocks the ability of this phosphatase to dephosphorylate SAPK.

The glycolytic enzyme enolase (EC 4.2.1.11) exists as dimers formed from three structurally related subunits alpha, beta, and gamma, encoded by separate genes. The gene encoding the beta-subunit is expressed only in striated muscles. We have previously shown that the beta-enolase gene belongs to a small subset of muscle-specific genes showing transcriptional activity in cultured myoblasts, prior to withdrawal from the cell cycle. An increase in the level of beta-enolase mRNA occurs during terminal differentiation of myoblasts. To investigate the mechanisms underlying this increase, we have simultaneously estimated, under steady state conditions, the rate of synthesis and the stability of beta-enolase mRNA in proliferating C2.7 myoblasts as well as in differentiating myotubes. The method used is based on the isolation of newly synthesized RNA from the total RNA pool, following pulse-labeling of intact cells in the presence of 4-thiouridine. The results described here demonstrate a coordinate increase in newly synthesized and total beta-enolase mRNA, while the mRNA half-life, about 4 hr, remains unchanged in the course of terminal differentiation. The expression of the gene for insulin-like growth factor-II (IGF-II), a major positive regulator of myogenesis, was analyzed using the same approach. It is concluded that the up-regulation of beta-enolase as well as IGF-II gene expression in differentiating muscle cells reflects an increased rate of entry of newly synthesized mRNAs into the general pool of transcripts without changes in their respective half-lives.

Insulin-like growth factor-II (IGF-II) mRNA exists as multiple transcript size classes, such as 6.0, 5.3, 4.9, 3.2, and 2.2 kb mRNAs in various human tissues. Three different promoters, 2 different polyadenylation sites, and alternative splicing are involved in producing these multiple transcripts. Initiation of transcription at the 3 different promoters results in multiple mRNAs which contain identical coding regions but different 5'-untranslated regions (5'-UTRs). The first promoter is thought to direct expression of 5.3 kb mRNA in adult human liver. The second promoter region directs expression of 6.0, 3.2, and 2.2 kb mRNAs in human fetal tissues and several adult nonliver tissues. The third promoter specifies transcription of a 4.9 kb mRNA in various tissues. We isolated and sequenced a cDNA clone (pIGF-II-1-70) from a human placental cDNA library, which contains the IGF-II coding region and the 5'-UTR associated with the third promoter. By using a 5'-UTR-specific probe from the clone, we found that this third 5'-UTR is contained in the IGF-II mRNA of 2.2 kb and is absent in the 3.2 kb IGF-II mRNA. We also found an 0.9 kb transcript expressed in placenta, which hybridized strongly to the third 5'-UTR specific probe but not to IGF-II coding region probes. This finding might indicate the existence of an mRNA encoding an IGF-II-associated peptide.

Sampling in an incompletely mixed effluent was studied with the aid of a hydraulic model. The effluent was the discharge from a major wastewater treatment facility. Residual chemicals from the chlorination/dechlorination process were of concern. Concentration time-series data obtained in a 1:12 scale model were used to determine the best sampling location and duration and to predict expected sampling errors relative to spatially and temporally averaged concentration values. Some general concepts for sampler placement in not fully homogeneous streams were derived. Spatial concentration gradients have to be determined to select number and location of multiple sampling points. When concentrations fluctuate in time due to process controls, the flow patterns and velocity distributions have to be known to select representative sampling points. For example, representative sampling within a separated flow region requires carefully selected probe placement and long sampling durations. Physical model simulations can provide the information on flow and concentration time series that are necessary to select the sampling strategy.

BACKGROUND:In early 1990 fluconazole was introduced as a prophylactic antifungal agent after bone marrow transplantation. During the same year Candida krusei emerged as the chief candida pathogen among patients with bone marrow transplants.METHODS:To determine whether there was a correlation between the introduction of fluconazole and the increased incidence of C. krusei, we conducted a retrospective study based on the medical, mycologic, and autopsy records of all adult inpatients who had undergone bone marrow transplantation (n = 296) or who had leukemia (n = 167) at the study center during 1989 and 1990.RESULTS:The 84 patients who received antifungal prophylaxis with fluconazole had a sevenfold greater frequency of C. krusei infection than the 335 patients who did not receive fluconazole (8.3 percent vs. 1.2 percent, P = 0.002), despite having a lower frequency of disseminated C. albicans and C. tropicalis infections (0 vs. 6.0 percent, P = 0.02). Ten of the 11 C. krusei infections were controlled by a combination of amphotericin B and flucytosine. Colonization by C. krusei was found in 40.5 percent of the patients who received fluconazole but in only 16.7 percent of those who did not receive it (P less than 0.0001). Colonization was independently associated with the prophylactic use of both fluconazole (odds ratio, 3.50; P less than 0.001) and norfloxacin (odds ratio, 2.53; P = 0.04). C. krusei was not susceptible to fluconazole in vitro.CONCLUSIONS:In patients at high risk for disseminated candida infections, suppression of bacterial flora and the more common candida pathogens may permit some less pathogenic, but natively resistant candida species, such as C. krusei, to emerge as systemic pathogens.

The authors studied the biodisposition and hepatobiliary transport of two potential magnetic resonance imaging contrast agents, the iron (III) chelates of N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) and diastereomeric N,N'-ethylenebis(2-hydroxyphenylglycine) (EHPG). Fecal and urinary excretion (mean +/- standard deviation), respectively, of FE-59 over 7 days in rats given tracer doses of the contrast agents were 67% +/- 2% and 22% +/- 1% for Fe-59-HBED and 22% +/- 2% and 26% +/- 5% for Fe-59-EHPG. In bile duct-cannulated rats given 0.05-mmol/kg doses labeled with Fe-59, 52% +/-8% of Fe-59 from Fe-59-HBED but only 17% +/-3% of that from Fe-59-EHPG was excreted into the bile within 90 minutes. Bromosulfophthalein, but not taurocholate or oxyphenonium. was shown to compete with both agents for hepatobiliary transport. Enterohepatic recirculation of both agents was less than 1%.

Flow and mixing in a wastewater dechlorination basin are investigated by tracing dye concentrations in a 1:12 scale sectional model and in a fullscale installation. The injection of an SO2 solution that neutralizes residual chlorine is simulated by dye. Time series measurements of dye concentrations are obtained in the model and in the full‐scale basin. Mean dye concentrations and standard deviations of concentration fluctuations measured throughout the model and the full‐scale basin compare favorably. Both show incomplete mixing of SO2 solution and residual chlorine. The model is then used to develop a substantially improved basin design by relocation of SO2 injection and addition of a columnar wall to redistribute the flow and create small‐scale turbulence. The modifications can be implemented without interruption of wastewater treatment. The performance of the improved design is again validated in the full‐scale operation. The study shows that existing problems of mixing and flow processes can be explored and corrected efficiently and effectively using a scale model. It is recommended that this method be used for the study of novel designs before construction.