To identify genes that could serve as targets for novel cancer therapeutics, we used a bioinformatic analysis of microarray data comparing gene expression between normal and tumor-derived primary ...human tissues. From this approach, we have found that maternal embryonic leucine zipper kinase (Melk), a member of the AMP serine/threonine kinase family, exhibits multiple features consistent with the potential utility of this gene as an anticancer target. An oligonucleotide microarray analysis of multiple human tumor samples and cell lines suggests that Melk expression is frequently elevated in cancer relative to normal tissues, a pattern confirmed by quantitative reverse transcription-PCR and Western blotting of selected primary tumor samples. In situ hybridization localized Melk expression to malignant epithelial cells in 96%, 23%, and 13% of colorectal, lung, and ovarian tissue tumor samples, respectively. Expression of this gene is also elevated in spontaneous tumors derived from the ApcMin and Apc1638N murine models of intestinal tumorigenesis. To begin addressing whether Melk is relevant for tumorigenesis, RNA interference-mediated silencing within human and murine tumor cell lines was done. We show that Melk knockdown decreases proliferation and anchorage-independent growth in vitro as well as tumor growth in a xenograft model. Together, these results suggest that Melk may provide a growth advantage for neoplastic cells and, therefore, inactivation may be therapeutically beneficial.
We report the measurement of the parity-violating asymmetry for the inelastic scattering of electrons from the proton, at $Q^2 = 0.082$ GeV$^2$ and $ W = 2.23$ GeV, above the resonance region. The ...result $A_{\rm Inel} = - 13.5 \pm 2.0 ({\rm stat}) \pm 3.9 ({\rm syst})$ ppm agrees with theoretical calculations, and helps to validate the modeling of the $\gamma Z$ interference structure functions $F_1^{\gamma Z}$ and $F_2^{\gamma Z}$ used in those calculations, which are also used for determination of the two-boson exchange box diagram ($\Box_{\gamma Z}$) contribution to parity-violating elastic scattering measurements. A positive parity-violating asymmetry for inclusive $\pi^-$ production was observed, as well as positive beam-normal single-spin asymmetry for scattered electrons and a negative beam-normal single-spin asymmetry for inclusive $\pi^-$ production.
To describe implementation of diabetes and hypertension program in rural Dominican Republic (DR), and report six years of quality improvement process and health outcomes.
Dominican teams at two ...clinics are supported by Chronic Care International with: supervision and continuing education, electronic database, diabetes and hypertension protocols, medications, self-management education materials, behavior change techniques, and equipment and testing supplies (e.g., HbA1c, lipids, blood pressure, BMI). A monthly dashboard for care processes and health outcomes guides problem solving and goal setting. Results were analyzed for quality improvement reports and by fitting the clinical data to random-effects linear models.
1191 adults were enrolled in the program at two clinics (44% men, baseline means: 56.4 years, BMI 27.4 kg/m2, HbA1c 8.8% (73 mmol/mol), BP 133/81 mmHg). Data show steady growth in clinic populations reaching capacity. Protocols for comprehensive foot examinations, BP and HbA1c assessments, and proportions reaching quality measures improved over time, especially after clinic goal setting. Modeling of BP, BMI and HbA1c values revealed important differences in outcomes by clinic over time.
Improvements in process and health outcomes are attainable in rural DR when medical teams have support and access to data. Scalability and sustainability are continuing goals.
Efficient single nucleotide polymorphism (SNP) genotyping methods are necessary to accomplish many current gene discovery goals. A crucial element in large-scale SNP genotyping is the number of ...individual biochemical reactions that must be performed. An efficient method that can be used to simultaneously amplify a set of genetic loci across a genome with high reliability can provide a valuable tool for large-scale SNP genotyping studies. In this paper we describe and characterize a method that addresses this goal. We have developed a strategy for reducing genome complexity by using degenerate oligonucleotide primer (DOP)-PCR and applied this strategy to SNP genotyping in three complex eukaryotic genomes; human, mouse, and Arabidopsis thaliana. Using a single DOP-PCR primer, SNP loci spread throughout a genome can be amplified and accurately genotyped directly from a DOP-PCR product mixture. DOP-PCRs are extremely reproducible. The DOP-PCR method is transferable to many species of interest. Finally, we describe an in silico approach that can effectively predict the SNP loci amplified in a given DOP-PCR, permitting the design of an efficient set of reactions for large-scale, genome-wide SNP studies.
Saprolite is a form of weathered bedrock that is commonly used as the host material at waste disposal sites in the Southeastern Piedmont. However, estimating the unsaturated hydraulic and transport ...properties of saprolite is difficult due to saprolite's low permeability. We demonstrate the use of short‐duration fluid irrigation pulses for maintaining unsaturated conditions in intact saprolite columns. Concomitant Cl− tracer experiments demonstrate that irrigated waters moved through an effective volumetric porosity (0.038–0.108 cm3 cm−3) substantially less than the ambient water‐filled porosity (0.44 cm3 cm−3). We observed the unexpected result that irrigation‐induced pressure wave velocities (1983–3670 cm d−1) were ≈1000 times faster than tracer velocities (2.04–6.00 cm d−1). The relationship between pressure wave velocities and fluid velocities is described using kinematic wave theory, presented for four parametric representations (Brooks–Corey, van Genuchten–Mualem, Broadbridge–White, and the Galileo Number), that predicts fluid pressure velocities to be from approximately two to fifteen times faster than saprolite tracer velocities. None of the kinematic models was able to reproduce observed rapid pressure wave velocities. A hydraulic form of the advection–diffusion equation based on Richards' equation is presented that favorably predicts the shape of pressure response curves only when the kinematic velocity is ignored and the hydraulic diffusivity of the unsaturated saprolite is considered. Based on the advection–diffusion equation, diffusion‐dominated soil water pressure wave velocities should decrease with depth, eventually conforming with kinematic wave theory. Pressure pulse velocity monitoring may be an additional tool for estimating unsaturated hydraulic properties in low permeability media.
In this paper, the polarization response of a GaAs based two-photon absorption microcavity photodetector has been studied. The deviation in the dependence of the detector response from that of bulk ...GaAs is shown to be due to the birefringence of the cavity. A theoretical model based on the convolution of the cavity birefringence and the polarization dependence of two-photon absorption in GaAs is described and shown to match the measured polarization dependence of the microcavity detector very well.
Bilateral simultaneous venous sampling of ACTH from the inferior petrosal sinus is a reliable test for diagnosing Cushing's disease, but is not reliable for lateralizing ACTH-secreting pituitary ...adenomas. We reviewed 23 consecutive patients with Cushing's disease who underwent venous angiography of the cavernous and inferior petrosal sinuses followed by bilateral simultaneous venous sampling of ACTH in the inferior petrosal and cavernous sinuses. Venous drainage was bilaterally symmetric in 14 patients (61%) and asymmetric in 9 (39%). The most common asymmetric pattern (6 patients) was for blood from both cavernous sinuses to drain into the right inferior petrosal sinus, with no significant drainage into the left. Cavernous sinus sampling in 21 patients correctly lateralized the tumor in 12 cases of symmetric venous drainage, but in only 3 cases of asymmetric drainage. Inferior petrosal sinus sampling in all 23 patients correctly lateralized the tumor in 12 cases of symmetric drainage, but in only four cases of asymmetric drainage. Overall, venous sampling correctly lateralized 70% of the tumors. Incorrect lateralization in cases of asymmetric venous drainage is probably attributable to shunting of blood toward the side of dominant venous drainage. Our findings illustrate the need for venography in all patients undergoing venous sampling of ACTH because an understanding of the venous drainage patterns is essential to correctly interpret venous sampling data and warn physicians that the lateralization data may be incorrect or unreliable.
Some data suggest that the sodium-dependent, high-affinity L-glutamate (Glu) transport sites in forebrain are different from those in cerebellum. In the present study, sodium-dependent transport of ...L-3HGlu was characterized in cerebellum and cortex. In both cerebellar and cortical tissue, activity was enriched in synaptosomes. Approximately 100 excitatory amino acid analogues were tested as potential inhibitors of transport activity. Many of the compounds tested inhibited transport activity by < 65% at 1 mM and were not studied further. One group of compounds exhibited inhibition conforming to theoretical curves with Hill coefficients of 1 and were < 10-fold selective as inhibitors of transport activity. These included three of the putative endogenous substrates for transport: L-Glu, L-aspartate, and L-cysteate. Four of the compounds exhibited inhibition conforming to theoretical curves with Hill coefficients of 1 and were > 10-fold selective as inhibitors. These included beta-N-oxalyl-L-alpha,beta-diaminopropionate, alpha-methyl-DL-glutamate, (2S,1'S,2'S)-2-(carboxycyclopropyl)glycine, and (2S,1'S,2'S,3'S)-2-(2-carboxy-3-methoxymethylcyclopropyl)glycine. Data obtained with a few of the inhibitors were consistent with two sites in one or both of the brain regions. (2S,1'R,2'R)-2-(Carboxycyclopropyl)glycine (L-CCG-II) was identified as the most potent (IC50 = 5.5 microM) and selective (60-100-fold) inhibitor of transport activity in cerebellum. One of the potential endogenous substrates, L-homocysteate, was also a selective inhibitor of cerebellar transport activity. The data for inhibition of transport activity in cortex by both L-CCG-II and L-homocysteate were best fit to two sites. Kainate was equipotent as an inhibitor of transport activity, and in both brain regions the data for inhibition were best fit to two sites. The possibility that there are four subtypes of excitatory amino acid transport is discussed. Altering sodium and potassium levels affects cerebellar and cortical transport activity differently, suggesting that the differences extend to other recognition sites on these transporters.
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