Investigating groundwater flow between Edwards and Trinity aquifers in central Texas.

Understanding the nature of communication between aquifers can be challenging when using traditional physical and geochemical groundwater sampling approaches. This study uses two multiport wells completed within Edwards and Trinity aquifers in central Texas to determine the degree of groundwater inter-flow between adjacent aquifers. Potentiometric surfaces, hydraulic conductivities, and groundwater major ion concentrations and Sr isotope values were measured from multiple zones within three hydrostratigraphic units (Edwards and Upper and Middle Trinity aquifers). Physical and geochemical data from the multiport wells were combined with historical measurements of groundwater levels and geochemical compositions from the region to characterize groundwater flow and identify controls on the geochemical compositions of the Edwards and Trinity aquifers. Our results suggest that vertical groundwater flow between Edwards and Middle Trinity aquifers is likely limited by low permeability, evaporite-rich units within the Upper and Middle Trinity. Potentiometric surface levels in both aquifers vary with changes in wet vs. dry conditions, indicating that recharge to both aquifers occurs through distinct recharge areas. Geochemical compositions in the Edwards, Upper, and Middle Trinity aquifers are distinct and likely reflect groundwater interaction with different lithologies (e.g., carbonates, evaporites, and siliceous sediments) as opposed to mixing of groundwater between the aquifers. These results have implications for the management of these aquifers as they indicate that, under current conditions, pumping of either aquifer will likely not induce vertical cross-formational flow between the aquifers. Inter-flow between the Trinity and the Edwards aquifers, however, should be reevaluated as pumping patterns and hydrogeologic conditions change.

Serial changes in the expression of breast cancer-related proteins in response to neoadjuvant chemotherapy.

BACKGROUND: Tumour expression of cyclooxygenase-2 (COX-2), epidermal growth factor receptor (EGFR), erythroblastic leukaemia viral oncogene homologue-2 (ErbB2), Ki-67 and p53 in breast cancer are associated with poorer outcomes. We investigated in vivo changes of these proteins with neoadjuvant chemotherapy. PATIENTS AND METHODS: Four core biopsies were taken from 100 breast cancer patients at baseline, during and upon completion of neoadjuvant chemotherapy. Immunohistochemical expression of these proteins were evaluated and correlated with clinicopathological features, clinical response and progression-free survival (PFS). RESULTS: There was a statistically significant change from positivity to negativity in COX-2 expression with chemotherapy (P = 0.002), predominantly in clinical responders (P = 0.002). COX-2-positive tumours that remained positive had shorter PFS than those that turned negative. Estrogen receptor (ER)+ and COX-2+ tumours at baseline that remained COX-2+ fared worse than those that became COX-2 negative (PFS 27 versus 52 months, P = 0.002). No significant changes in IHC expression were observed for ER, progesterone receptor, ErbB2, EGFR, p53 or Ki67. CONCLUSIONS: Chemotherapy induced change in COX-2 expression from positivity to negativity predominantly among clinical responders and is associated with longer PFS. Interaction between COX-2 and ER was observed, suggesting that some hormone receptor-positive patients may benefit from combining COX-2 inhibition with hormonal therapy.

Detection of naturally occurring enteroviruses in waters using direct RT-PCR and integrated cell culture-RT-PCR.

Viruses detected by rapid molecular assays are not always infectious. In this study we compared enterovirus levels in natural waters using culture and reverse transcription-polymerase chain reaction (RT-PCR) techniques to determine whether molecular units of naturally occurring enteroviruses can be utilized to predict viral infectivity. Viruses were concentrated from 12 river water and effluent samples using 1 MDS filter-filtration and beef extract-elution. An integrated cell culture-RT-PCR (ICC-RT-PCR) was applied to the concentrates; and these waters contained up to 1.9 MPN of culturable (on BGM cells) viruses per litre (0.57 MPN/300 ml). Sample concentrates were also subjected to a direct 'molecular' approach using solvent-extraction, PEG-precipitation, and RNA-extraction before RT-PCR detection. The detection sensitivity of the direct RT-PCR was equivalent to 0.46 estimated (culturable) MPN/reaction, per 300 ml water. Two-thirds of the samples demonstrated consistent presence or absence of viruses by ICC-RT-PCR and direct RT-PCR. The direct RT-PCR approach resulted in over-estimation of naturally occurring infectious viruses as high as 91-fold in waters. Increased RT-PCR units may not reflect higher levels of culturable viruses in natural waters. The differences in virus levels detected by molecular and culture assays could be attributed to factors of volume of sample analyzed, different concentration schemes utilized that may affect the presence of residual inhibitors, and different stability exhibited by enterovirus strains/groups.

PXR, CAR and HNF4alpha genotypes and their association with pharmacokinetics and pharmacodynamics of docetaxel and doxorubicin in Asian patients.

Previously studied candidate genes have failed to account for inter-individual variability of docetaxel and doxorubicin disposition and effects. We genotyped the transcriptional regulators of CYP3A and ABCB1 in 101 breast cancer patients from 3 Asian ethnic groups, that is, Chinese, Malays and Indians, in correlation with the pharmacokinetics and pharmacodynamics of docetaxel and doxorubicin. While there was no ethnic difference in docetaxel and doxorubicin pharmacokinetics, ethnic difference in docetaxel- (ANOVA, P=0.001) and doxorubicin-induced (ANOVA, P=0.003) leukocyte suppression was observed, with Chinese and Indians experiencing greater degree of docetaxel-induced myelosuppression than Malays (Bonferroni, P=0.002, P=0.042), and Chinese experiencing greater degree of doxorubicin-induced myelosuppression than Malays and Indians (post hoc Bonferroni, P=0.024 and 0.025). Genotyping revealed both PXR and CAR to be well conserved; only a PXR 5'-untranslated region polymorphism (-24381A>C) and a silent CAR variant (Pro180Pro) were found at allele frequencies of 26 and 53%, respectively. Two non-synonymous variants were identified in HNF4alpha (Met49Val and Thr130Ile) at allele frequencies of 55 and 1%, respectively, with the Met49Val variant associated with slower neutrophil recovery in docetaxel-treated patients (ANOVA, P=0.046). Interactions were observed between HNF4alpha Met49Val and CAR Pro180Pro, with patients who were wild type for both variants experiencing least docetaxel-induced neutropenia (ANOVA, P=0.030). No other significant genotypic associations with pharmacokinetics or pharmacodynamics of either drug were found. The PXR-24381A>C variants were significantly more common in Indians compared to Chinese or Malays (32/18/21%, P=0.035) Inter-individual and inter-ethnic variations of docetaxel and doxorubicin pharmacokinetics or pharmacodynamics exist, but genotypic variability of the transcriptional regulators PAR, CAR and HNF4alpha cannot account for this variability.

Distinguishing androgen receptor agonists and antagonists: distinct mechanisms of activation by medroxyprogesterone acetate and dihydrotestosterone.

Natural and pharmacological androgen receptor (AR) ligands were tested for their ability to induce the AR NH2-terminal and carboxyl-terminal (N/C) interaction in a two-hybrid protein assay to determine whether N/C complex formation distinguishes in vivo AR agonists from antagonists. High-affinity agonists such as dihydrotestosterone, mibolerone, testosterone, and methyltrienolone at concentrations between 0.1 and 1 nM induce the N/C interaction more than 40-fold. The lower affinity anabolic steroids, oxandrolone and fluoxymesterone, require concentrations of 10-100 nM for up to 23-fold induction of the N/C interaction. However no N/C interaction was detected in the presence of the antagonists, hydroxyflutamide, cyproterone acetate, or RU56187, at concentrations up to 1 microM, or with 1 microM estradiol, progesterone, or medroxyprogesterone acetate; each of these steroids at 1-500 nM inhibited the dihydrotestosterone-induced N/C interaction, with medroxyprogesterone acetate being the most effective. In transient and stable cotransfection assays using the mouse mammary tumor virus reporter vector, all ligands displayed concentration-dependent AR agonist activity that paralleled induction of the N/C interaction, with antagonists and weaker agonists failing to induce the N/C interaction. AR dimerization and DNA binding in mobility shift assays and AR stabilization reflected, but were not dependent on, the N/C interaction. The results indicate that the N/C interaction facilitates agonist potency at low physiological ligand concentrations as detected in transcription, dimerization/DNA binding, and stabilization assays. However the N/C interaction is not required for agonist activity at sufficiently high ligand concentrations, nor does its inhibition imply antagonist activity.

Steroid requirement for androgen receptor dimerization and DNA binding. Modulation by intramolecular interactions between the NH2-terminal and steroid-binding domains.

Infection of Spodoptera frugiperda Sf9 insect cells with recombinant human androgen receptor (AR) baculovirus results in expression of a 118-kDa phosphoprotein that displays high affinity androgen binding and androgen-dependent targeting to the nucleus. Using the DNA mobility shift assay, specific in vitro binding of full-length AR to androgen response element DNA (ARE) requires intracellular hormone exposure. The ability of a variety of steroids to induce ARE binding paralleled their transcriptional potential. Certain antihormones, cyproterone acetate and RU486, promote ARE binding, but a pure antiandrogen, hydroxyflutamide, inhibits AR binding to ARE DNA. AR dimerization requires incubation of recombinant baculovirus-infected insect cells with androgen, but only when one or both components of the dimer contain the NH2-terminal domain. Based on the intensities of ARE binding and lack of binding to an ARE half-site, it appears that, unlike the glucocorticoid receptor, AR binds DNA primarily as a dimer. Thus, full-length baculovirus-expressed AR requires intracellular hormone exposure for dimerization and ARE binding to overcome inhibition imposed by the AR NH2-terminal domain. Antihormones with agonist activity promote dimerization and ARE binding, while a pure antiandrogen blocks AR DNA binding. It is concluded that intramolecular interactions between the NH2-terminal and steroid-binding domains are regulated by the specificity of hormone binding and modulate receptor dimerization and DNA binding.

Establishment and characterization of two nasopharyngeal carcinoma cell lines.

Two human nasopharyngeal carcinoma (NPC) cell lines have been established. One derived from a 64-year-old male, and the other from a 36-year-old female Chinese patient living in Taiwan. Both were keratinizing squamous cell carcinoma in nature and designated as NPC-TW039 and NPC-TW076. Both have been grown in culture system for more than 100 passages. Single cells from both cell lines could form colonies in 0.3% soft agar. In the nude mouse transplantation experiment, both cell lines could produce tumor mass with metastasis. The karyotypic analysis showed multiple chromosomal abnormality. The number of chromosomes ranged between 76 to 109 and 80 to 105 with an average of 98 and 95, respectively. The doubling time was 10.5 hours and 10.8 hours, respectively. The NPC-TW039 cell line has been subcloned and three subclones have been obtained. Ultrastructural studies from those two cell line, three subcloned cell lines and two transplanted tumor masses, all showed two types of morphology: the dark and light cells. This morphologic difference is probably derived from the different metabolic state, but not due to an artifact. Three oncogene probes have been used to check the oncogene expression; none of those five cell lines is positive. Similarly, six Epstein-Barr virus fragments have been labeled to hybridize with NPC cellular DNA preparations, results from the Southern blotting showed no detectable Epstein-Barr virus DNA sequence.