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Understanding how interacting particles approach thermal equilibrium is a major challenge of quantum simulators 1,2 . Unlocking the full potential of such systems towards this goal requires flexible initial state preparation, precise time evolution and extensive probes for final state characterization. Here we present a quantum simulator comprising 69 superconducting qubits that supports both universal quantum gates and high-fidelity analogue evolution, with performance beyond the reach of classical simulation in cross-entropy benchmarking experiments. This hybrid platform features more versatile measurement capabilities compared with analogue-only simulators, which we leverage here to reveal a coarsening-induced breakdown of Kibble-Zurek scaling predictions 3 in the XY model, as well as signatures of the classical Kosterlitz-Thouless phase transition 4 . Moreover, the digital gates enable precise energy control, allowing us to study the effects of the eigenstate thermalization hypothesis 5-7 in targeted parts of the eigenspectrum. We also demonstrate digital preparation of pairwise-entangled dimer states, and image the transport of energy and vorticity during subsequent thermalization in analogue evolution. These results establish the efficacy of superconducting analogue-digital quantum processors for preparing states across many-body spectra and unveiling their thermalization dynamics., Competing Interests: Competing interests: A provisional patent application has been submitted for the analogue calibration scheme, titled ‘High-accuracy calibration of an analog quantum simulator’ (application number 63/639,509). The listed inventors are T.I.A., J.A.G., D.A.A. and X.M. The other authors declare no competing interests., (© 2025. The Author(s).)

Undesired coupling to the surrounding environment destroys long-range correlations in quantum processors and hinders coherent evolution in the nominally available computational space. This noise is an outstanding challenge when leveraging the computation power of near-term quantum processors 1 . It has been shown that benchmarking random circuit sampling with cross-entropy benchmarking can provide an estimate of the effective size of the Hilbert space coherently available 2-8 . Nevertheless, quantum algorithms' outputs can be trivialized by noise, making them susceptible to classical computation spoofing. Here, by implementing an algorithm for random circuit sampling, we demonstrate experimentally that two phase transitions are observable with cross-entropy benchmarking, which we explain theoretically with a statistical model. The first is a dynamical transition as a function of the number of cycles and is the continuation of the anti-concentration point in the noiseless case. The second is a quantum phase transition controlled by the error per cycle; to identify it analytically and experimentally, we create a weak-link model, which allows us to vary the strength of the noise versus coherent evolution. Furthermore, by presenting a random circuit sampling experiment in the weak-noise phase with 67 qubits at 32 cycles, we demonstrate that the computational cost of our experiment is beyond the capabilities of existing classical supercomputers. Our experimental and theoretical work establishes the existence of transitions to a stable, computationally complex phase that is reachable with current quantum processors., (© 2024. The Author(s).)

Understanding universal aspects of quantum dynamics is an unresolved problem in statistical mechanics. In particular, the spin dynamics of the one-dimensional Heisenberg model were conjectured as to belong to the Kardar-Parisi-Zhang (KPZ) universality class based on the scaling of the infinite-temperature spin-spin correlation function. In a chain of 46 superconducting qubits, we studied the probability distribution of the magnetization transferred across the chain's center, [Formula: see text]. The first two moments of [Formula: see text] show superdiffusive behavior, a hallmark of KPZ universality. However, the third and fourth moments ruled out the KPZ conjecture and allow for evaluating other theories. Our results highlight the importance of studying higher moments in determining dynamic universality classes and provide insights into universal behavior in quantum systems.

Engineered dissipative reservoirs have the potential to steer many-body quantum systems toward correlated steady states useful for quantum simulation of high-temperature superconductivity or quantum magnetism. Using up to 49 superconducting qubits, we prepared low-energy states of the transverse-field Ising model through coupling to dissipative auxiliary qubits. In one dimension, we observed long-range quantum correlations and a ground-state fidelity of 0.86 for 18 qubits at the critical point. In two dimensions, we found mutual information that extends beyond nearest neighbors. Lastly, by coupling the system to auxiliaries emulating reservoirs with different chemical potentials, we explored transport in the quantum Heisenberg model. Our results establish engineered dissipation as a scalable alternative to unitary evolution for preparing entangled many-body states on noisy quantum processors.

Systems of correlated particles appear in many fields of modern science and represent some of the most intractable computational problems in nature. The computational challenge in these systems arises when interactions become comparable to other energy scales, which makes the state of each particle depend on all other particles 1 . The lack of general solutions for the three-body problem and acceptable theory for strongly correlated electrons shows that our understanding of correlated systems fades when the particle number or the interaction strength increases. One of the hallmarks of interacting systems is the formation of multiparticle bound states 2-9 . Here we develop a high-fidelity parameterizable fSim gate and implement the periodic quantum circuit of the spin-½ XXZ model in a ring of 24 superconducting qubits. We study the propagation of these excitations and observe their bound nature for up to five photons. We devise a phase-sensitive method for constructing the few-body spectrum of the bound states and extract their pseudo-charge by introducing a synthetic flux. By introducing interactions between the ring and additional qubits, we observe an unexpected resilience of the bound states to integrability breaking. This finding goes against the idea that bound states in non-integrable systems are unstable when their energies overlap with the continuum spectrum. Our work provides experimental evidence for bound states of interacting photons and discovers their stability beyond the integrability limit., (© 2022. The Author(s).)

Inherent symmetry of a quantum system may protect its otherwise fragile states. Leveraging such protection requires testing its robustness against uncontrolled environmental interactions. Using 47 superconducting qubits, we implement the one-dimensional kicked Ising model, which exhibits nonlocal Majorana edge modes (MEMs) with [Formula: see text] parity symmetry. We find that any multiqubit Pauli operator overlapping with the MEMs exhibits a uniform late-time decay rate comparable to single-qubit relaxation rates, irrespective of its size or composition. This characteristic allows us to accurately reconstruct the exponentially localized spatial profiles of the MEMs. Furthermore, the MEMs are found to be resilient against certain symmetry-breaking noise owing to a prethermalization mechanism. Our work elucidates the complex interplay between noise and symmetry-protected edge modes in a solid-state environment.

We report the characteristics of the strained In 0.65 Ga 0.35 As triple quantum well (QW) diode lasers grown by metalorganic vapor phase epitaxy (MOVPE) on lattice-mismatched substrates such as GaAs or Si, by utilizing InP metamorphic buffer layers (MBLs) in conjunction with InAs nanostructure-based dislocation filters. As the lattice-mismatch between the substrate and InP MBL increases, higher threshold current densities and lower slope efficiencies were observed, together with higher temperature sensitivities for the threshold current and slope efficiency. Structural analysis performed by both high-resolution X-ray diffraction (HR-XRD) and transmission electron microscopy indicates graded and/or rougher QW interfaces within the active region grown on the mismatched substrate, which accounts for the observed devices characteristics.

This study examined concordances of cancer patients' received and caregivers' provided support and dyadic relationship quality, and their predictive utility in prospective psychological distress and well-being. A total of 83 Chinese cancer patient-caregiver dyads were recruited in two government-funded hospitals in Hong Kong. Participants reported received (patient)/provided (caregiver) emotional and instrumental support and dyadic relationship quality within 6 months after diagnosis (T1), and anxiety and depressive symptoms, positive affect and life satisfaction at both T1 and 6-month follow-up (T2). We hypothesised that concordances at T1 would predict lower psychological distress and higher psychological well-being among both patients and caregivers at T2. Concordances were indicated by Gwet's AC 2 scores (possible range = -1.00 to 1.00) and as follows: emotional support: M = 0.92, SD = 0.12, range = 0.25-1.00; instrumental support: M = 0.92, SD = 0.16, range = 0.08-1.00; and relationship quality: M = 0.63, SD = 0.27, range = -0.31 to 1.00. Hierarchical multiple regressions revealed that T1 concordances of perceived emotional and instrumental support and dyadic relationship quality positively predicted T2 anxiety symptoms [F(9, 74) = 6.725, ∆R 2  = .031, p < .001)] and state positive affect [F(9, 74) = 3.436, ∆R 2  = .042, p = .001)], whereas inversely predicted T2 depressive symptoms [F(9, 74) = 4.189, ∆R 2  = .042, p < .01)]. Significant associations were found only among caregivers, but not patients., (© 2018 John Wiley & Sons Ltd.)

Danggui Buxue Tang, an ancient Chinese herbal decoction containing Astragali Radix and Angelicae Sinensis Radix at the weight ratio of 5:1, is used to mitigate menopausal syndromes in women. The pharmacological properties of Danggui Buxue Tang have been illustrated in bone development, blood enhancement, and immune stimulation. Here, we extended the possible pharmacological role of Danggui Buxue Tang in cardiovascular function. In cultured human umbilical vein endothelial cells, the application of Danggui Buxue Tang induced the release of nitric oxide and the phosphorylation of endothelial nitric oxide synthase and Akt kinase in time- and dose-dependent manners. The robust activation of nitric oxide signaling, however, required the boiling of Astragali Radix and Angelicae Sinensis Radix together, i.e., as Danggui Buxue Tang instead of other herbal extracts. The Danggui Buxue Tang-induced phosphorylation of endothelial nitric oxide synthase and Akt kinase in human umbilical vein endothelial cells were fully blocked by treatment with an endothelial nitric oxide synthase inhibitor (L-NAME), a PI3K/Akt inhibitor (LY294002), and a Ca(2+) chelator (BAPTA-AM). In parallel, the blockage of endothelial nitric oxide synthase and Akt activation subsequently fully abolished the Danggui Buxue Tang-induced nitric oxide production., (Georg Thieme Verlag KG Stuttgart · New York.)

Introduction: Misinterpretation of abbreviations by healthcare professionals has been reported to compromise patient safety. This study was done to determine the prevalence of abbreviations usage among medical doctors and nurses and their ability to interpret commonly used abbreviations in medical practice., Methods: Seventy-seven medical doctors and eighty nurses answered a self-administered questionnaire designed to capture demographic data and information regarding abbreviation use in medical practice. Comparisons were made between doctors and nurses with regards to frequency and reasons for using abbreviations; from where abbreviations were learned; frequency of encountering abbreviations in medical practice; prevalence of medical errors due to misinterpretation of abbreviations; and their ability to correctly interpret commonly used abbreviations., Results: The use of abbreviations was highly prevalent among doctors and nurses. Time saving, avoidance of writing sentences in full and convenience, were the main reasons for using abbreviations. Doctors learned abbreviations from fellow doctors while nurses learned from fellow nurses and doctors. More doctors than nurses reported encountering abbreviations. Both groups reported no difficulties in interpreting abbreviations although nurses reported often resorting to guesswork. Both groups felt abbreviations were necessary and an acceptable part of work. Doctors outperformed nurses in correctly interpreting commonly used standard and non-standard abbreviations., Conclusion: The use of standard and non-standard abbreviation in clinical practice by doctors and nurses was highly prevalent. Significant variability in interpretation of abbreviations exists between doctors and nurses.

This article investigates the impact of Saharan dust on the development of tropical cyclones in the Atlantic. A global data assimilation and forecast system, the NASA GEOS-5, is used to assimilate all satellite and conventional data sets used operationally for numerical weather prediction. In addition, this new GEOS-5 version includes assimilation of aerosol optical depth from the Moderate Resolution Imaging Spectroradiometer. The analysis so obtained comprises atmospheric quantities and a realistic 3-D aerosol and cloud distribution, consistent with the meteorology and validated against Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation and CloudSat data. These improved analyses are used to initialize GEOS-5 forecasts, explicitly accounting for aerosol direct radiative effects and their impact on the atmospheric dynamics. Parallel simulations with/without aerosol radiative effects show that effects of dust on static stability increase with time, becoming highly significant after day 5 and producing an environment less favorable to tropical cyclogenesis.

Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is expressed in the epithelial cells of a wide range of organs/tissues from which most cancers are derived. Although accumulating reports have indicated the association of cancer incidence with genetic variations in CFTR gene, the exact role of CFTR in cancer development and the possible underlying mechanism have not been elucidated. Here, we report that CFTR expression is significantly decreased in both prostate cancer cell lines and human prostate cancer tissue samples. Overexpression of CFTR in prostate cancer cell lines suppresses tumor progression (cell growth, adhesion and migration), whereas knockdown of CFTR leads to enhanced malignancies both in vitro and in vivo. In addition, we demonstrate that CFTR knockdown-enhanced cell proliferation, cell invasion and migration are significantly reversed by antibodies against either urokinase plasminogen activator (uPA) or uPA receptor (uPAR), which are known to be involved in various malignant traits of cancer development. More interestingly, overexpression of CFTR suppresses uPA by upregulating the recently described tumor suppressor microRNA-193b (miR-193b), and overexpression of pre-miR-193b significantly reverses CFTR knockdown-enhanced malignant phenotype and abrogates elevated uPA activity in prostate cancer cell line. Finally, we show that CFTR gene transfer results in significant tumor repression in prostate cancer xenografts in vivo. Taken together, the present study has demonstrated a previously undefined tumor-suppressing role of CFTR and its involvement in regulation of miR-193b in prostate cancer development.

Minichromosome maintenance (MCM) proteins 2-7 are important in DNA replication licensing. Functional roles beyond licensing are speculated. In addition, significances in medulloblastoma (MB) remain unclear. In this study, we showed the frequent deregulation of MCM2 and MCM3 expression in 7 MB cell lines and 31 clinical samples. Moreover, DAOY and ONS76 and the clinical samples expressed elevated MCM7 transcripts with genomic gain of the gene. Immunopositivity restricted to tumor cells was found in 41, 37 and 53 out of 73 MB cases for MCM2, MCM3 and MCM7, respectively. High-MCM3 expression was associated with poor prognosis. Knockdowns of these MCMs significantly inhibited anchorage-dependent and -independent MB cell growth. The inhibition of MCM3 expression by small interfering RNA knockdown was related to G1 arrest with reduced cyclin A expression, whereas the MCM2- and MCM7-knocked-down cells arrested at G2/M with increased cyclin A expression. Interestingly, we demonstrated the links of these MCMs with cell migration and invasion using wound-healing and Transwell migration/invasion assays. Exogenous overexpression of MCM2, MCM3 and MCM7 increased anchorage-independent cell growth, and also cell migration and invasion capabilities in MB cells. The knockdown reduced the number of filopodial cells and the cells with intense stress fibers by blocking cdc42 and Rho activation. Taken together, deregulation of MCM2, MCM3 and MCM7 expression might be involved in MB tumorigenesis and we revealed undefined roles of these MCMs in control of MB cell migration and invasion.

G-protein-coupled receptor-30 (GPR30) shows estrogen-binding affinity and mediates non-genomic signaling of estrogen to regulate cell growth. We here showed for the first time, in contrast to the reported promoting action of GPR30 on the growth of breast and ovarian cancer cells, that activation of GPR30 by the receptor-specific, non-estrogenic ligand G-1 inhibited the growth of androgen-dependent and androgen-independent prostate cancer (PCa) cells in vitro and PC-3 xenografts in vivo. However, G-1 elicited no growth or histological changes in the prostates of intact mice and did not inhibit growth in quiescent BPH-1, an immortalized benign prostatic epithelial cell line. Treatment of PC-3 cells with G-1 induced cell-cycle arrest at the G(2) phase and reduced the expression of G(2)-checkpoint regulators (cyclin-A2, cyclin-B1, cdc25c, and cdc2) and phosphorylation of their common transcriptional regulator NF-YA in PC-3 cells. With extensive use of siRNA-knockdown experiments and the MEK inhibitor PD98059 in this study, we dissected the mechanism underlying G-1-induced inhibition of PC-3 cell growth, which was mediated through GPR30, followed by sustained activation of Erk1/2 and a c-jun/c-fos-dependent upregulation of p21, resulting in the arrest of PC-3 growth at the G(2) phase. The discovery of this signaling pathway lays the foundation for future development of GPR30-based therapies for PCa.

Monitoring of trace impurities in electroplating bath is needed to meet EU requirements for WEEE and RoHS and for quality control of electrodeposits. Methods using IC and 100% aqueous CE buffer were found producing non-repeatable results attributed to interference of surfactants and major methanesulphonate anion. A new CE buffer containing 1.5mM tetraethylenepentaamine, 3mM 1,3,5-benzenetricarboxylic acid and 15 mM Tris in 20% (v/v) methanol at pH=8.4 was shown to enhance the separation window, reduce interaction between buffer and bath constituents, and give satisfactory repeatability with baseline separation for 14 organic and inorganic anions within 14 min, good repeatability for migration time (0.32-0.57% RSD), satisfactory peak area and peak height (2.9-4.5 and 3-4.7% respectively), low detection limit (S/N=2, 20-150 ppb), and wide working ranges (0.1-100 ppm). The CE buffer with 20% (v/v) methanol has demonstrated its capability for identifying anion impurities causing problem in aged tin bath and the use of only 10-fold dilution to produce reliable results for quality assessment in plating bath containing high surfactant additives., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Over 194 million people suffer from diabetes worldwide. The improper control of diabetes may result in diabetic foot ulcer or even amputation. Herbal medicine provides a means for treating diabetic foot ulcers for a large population in developing countries. The wound healing-enhancing activities of the principal herbs, Radix Astragali (RA) and Radix Rehmanniae (RR) in two clinically efficacious Chinese herbal formulae were studied in primary fibroblasts from diabetic foot ulcer patients. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that RA and RR significantly enhanced the viability of fibroblasts isolated from foot ulcers of diabetic patients, even from those with no response to insulin treatment. The results in this study indicate that fibroblast viability enhancement effects of RA and RR likely underlie the healing effects of F1 and F2 in diabetic foot ulcers., ((c) 2009 John Wiley & Sons, Ltd.)

Ethnopharmacological Relevance: Radix Rehmanniae (RR) has a very long history of usage in traditional Chinese medicine and is usually one of the principal herb found in many herbal formulae used in diabetic foot ulcer., Aim of the Study: RR aqueous extract was investigated for its wound healing effects in a diabetic foot ulcer rat model and its detailed mechanism of actions., Materials and Methods: A previously established diabetic foot ulcer rat model was used to assess the effect of RR extract on wound area reduction, tissue regeneration and angiogenesis. Carrageenan-induced inflammation rat model was used for inflammation study; and diabetic control was evaluated using a neonatal streptozotocin-induced diabetic rat model., Results: In the RR treated group, a trend of reduction of the wound area was observed from days 8 to 18 and a significant difference (as compared with control group) was found on day 8. The ulcer healing effect of RR extract was further supported by better developed scars and epithelialization as well as good formation of capillaries with enhanced VEGF expression. Carrageenan-induced inflammation was also significantly alleviated with RR extract., Conclusions: Our results demonstrated for the first time that Radix Rehmanniae was effective in promoting diabetic foot ulcer healing in rats through the processes of tissue regeneration, angiogenesis and inflammation control, but not glycemia control. The present study provided scientific basis to support the traditional use of Radix Rehmanniae in diabetic foot ulcer.

Fructus Corni, Fructus Schisandrae Chinensis, Poria, Rhizoma Alismatis and Rhizoma Dioscoreae are commonly used in traditional Chinese medicine for diabetes treatment. They are also the component herbs of an antidiabetic foot ulcer formula with demonstrated clinical efficacy. Although some of these herbal extracts were previously shown to possess in vivo antidiabetic effects (i.e. lowering blood glucose levels), the underlying mechanisms remain elusive. The objective of this study is to investigate the possible antidiabetic mechanisms of these individual herbs, using a systematic study platform which includes four in vitro tissue models: glucose absorption into intestinal brush border membrane vesicles (BBMV), gluconeogenesis by rat hepatoma cell line H4IIE, glucose uptake by human skin fibroblasts cell line Hs68 and mouse adipocytes 3T3-L1. All tested herbs showed significant in vitro antidiabetic effects in at least two models. Fructus Schisandrae Chinensis, Poria, Rhizoma Alismatis and Rhizoma Dioscoreae showed significant inhibitory effects in the BBMV glucose uptake assay. All tested herbs showed significant stimulatory effects to the glucose uptake of Hs68 and 3T3-L1 cells, except Poria and Rhizoma Dioscoreae which were not effective to Hs68 and 3T3-L1 respectively. However, none of the tested herbs inhibited hepatic gluconeogenesis. In conclusion, the five herbs exhibited distinct antidiabetic mechanisms in vitro and hence our investigations provided scientific evidence to support the traditional usage of these herbs for diabetic treatment in medicinal formulae., ((c) 2008 John Wiley & Sons, Ltd.)

Cortex Moutan (CM, root bark of Paeonia suffruticosa Andr.) is one of the common herbs found in anti-diabetic traditional Chinese medicine formulae. To study the potential anti-diabetic mechanisms of CM, four in vitro models (intestinal brush border membrane vesicles (BBMV), rat hepatoma cell line H4IIE, human skin fibroblasts cell line Hs68 and mouse adipocytes 3T3-L1) were used. CM showed significant in vitro anti-diabetic effects by inhibiting glucose uptake of BBMV and enhancing glucose uptake into Hs68 and 3T3-L1 cells. Using bioassay-guided fractionation, paeonol was confirmed to be one of the active constituents for inhibiting BBMV glucose uptake. With neonatal-streptozotocin diabetic rats, paeonol (200 and 400mg/kgbody wt.) was found to improve oral glucose tolerance in vivo. To the best of our knowledge, this is the first report on the anti-diabetic effect of paeonol.

A previous study conducted in Taiwan found a 100% association between HLA-B*1502 allele and carbamazepine-induced Stevens-Johnson syndrome (SJS) in Han Chinese subjects, with an extremely high odds ratio compared with carbamazepine-tolerant subjects (odds ratio = 2,504). We examined this association in 24 Hong Kong Han Chinese subjects who had cutaneous adverse reactions induced by different antiepileptic drugs (AEDs). They were matched with 48 AED-tolerant controls. HLA-B*1502 was associated with severe cutaneous reactions (SCR) induced by AEDs, which included carbamazepine, phenytoin, and lamotrigine (p = 0.001, odds ratio = 17.6), but was not associated with maculopapular exanthema (MPE) (p = 0.32). Further studies in larger samples of ethnically matched subjects should be conducted to confirm the findings. Identification of genetic polymorphisms predisposing to development of AED-induced SCR offers the possibility of avoiding these high-risk drugs in genetically susceptible individuals.

Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus-associated disease with high prevalence in Southern Chinese. Using multiparametric flow cytometry, we identified significant expansions of circulating naïve and memory CD4+CD25(high) T cells in 56 NPC patients compared with healthy age- and sex-matched controls. These were regulatory T cells (Treg), as they overexpressed Foxp3 and GITR, and demonstrated enhanced suppressive activities against autologous CD4+CD25- T-cell proliferation in functional studies on five patients. Abundant intraepithelial infiltrations of Treg with very high levels of Foxp3 expression and absence of CCR7 expression were also detected in five primary tumours. Our current study is the first to demonstrate an expansion of functional Treg in the circulation of NPC patients and the presence of infiltrating Treg in the tumour microenvironment. As Treg may play an important role in suppressing antitumour immunity, our findings provide critical insights for clinical management of NPC.

Complications of diabetes impose major public health burdens worldwide. The positive effect of a Radix Astragali-based herbal preparation on healing diabetic foot ulcers in patients has been reported. Formula 1 is also referred as the 'Herbal drink to strengthen muscle and control swelling'. This formula contains six Chinese medical herbs, including Radix Astragali, Radix Rehmanniae, Rhizoma Smilacis Chinensis, Rhizoma Atractylodis Macrocephalae, Radix Polygoni Multiflori Preparata, and Radix Stephania Tetrandrae. Three of these herbs (Radix Astragali, Radix Rehmanniae, Rhizoma Atractylodis Macrocephalae) are commonly used in different anti-diabetic formulae of Chinese medicine. The objective of the current study is to use an interdisciplinary approach to test the hypothesis that Formula 1 and its components influence tissue and systemic glucose homeostasis. In vitro and in vivo models have been established including: (1) glucose absorption into intestinal brush border membrane vesicles (BBMV); (2) gluconeogenesis by H4IIE hepatoma cells; (3) glucose uptake by 3T3-L1 adipocytes and Hs68 skin fibroblasts; (4) normalization of glycaemic control in a diabetic rat model. The results of in vitro studies indicated that all herbal extracts can modify cellular glucose homeostasis. Since Formula 1 and Rhizoma Smilacis Chinensis extracts demonstrated potent effects on modifying glucose homeostasis in multiple tissues in vitro, they were further studied for their anti-diabetic activities in vivo using a streptozotocin (STZ)-induced diabetic rat model. The results showed that Formula 1 and Rhizoma Smilacis Chinensis extracts did not significantly improve oral glucose tolerance or basal glycaemia in diabetic rats. In conclusion, the anti-diabetic foot ulcer Formula 1 contains ingredients active in modifying tissue glucose homeostasis in vitro but these biological activities could not be associated with improved glycaemic control of diabetes in vivo.

We performed a retrospective analysis on the human leukocyte antigen (HLA) data of 53 consecutive Chinese patients with high-risk childhood acute lymphoblastic leukemia (ALL) diagnosed from 1989 to 2003. A significantly higher frequency of HLA-B67 in the male relapse group of patients [OR, 23.08; 95% CI, 5.31-100.36; p = 0.0042; for statistical significance after Bonferroni correction (Bc) p (Bc) < 0.0083] was identified after Bonferroni correction. Although not surviving the Bonferroni correction, gender effects on the association were also observed with HLA-A11, HLA-A32, HLA-A33, and HLA-B22, which were however more prevalent in the female patients and particularly those developing relapse. Two patients with HLA-A29 and HLA-B7 revealed significantly shortened survivals, suggestive of their potential prognostic impacts. Notably, for the first time, we found a significant correlation of leukocyte count with HLA types, where HLA-A33 (p = 0.006) or HLA-B17 (p < 0.001) signifies higher leukocytosis at presentation. Taken together, our findings support the involvement of HLA in Chinese high-risk childhood ALL.

Cation determination is important for quality control of beverage products. To determine a large group simultaneously, a capillary electrophoresis procedure is developed with indirect UV at 214 nm in a three-complex buffer system (10 mM N,N-dimethylbenzylamine (DBA), 8 mM lactic acid and 2 mM 18-crown-6) with good mobility matching with desired cations. Under optimized conditions with pH adjusted to 4.65, a baseline separation is achieved for 14 cations (Rb(+), NH(4)(+), K(+), Ca(2+), Na(+), Mg(2+), Mn(2+), Co(2+), Fe(2+), Cd(2+), Cr(3+), Ni(2+), Zn(2+) and Cu(2+)) within 7 min using an uncoated silica column. To cover ng/l to mug/l range, both hydrostatic and electrokinetic sampling are studied, showing working ranges within (0.05-50)/(0.005-2) microg/l and detection limits (13-78)/(1.4-10) ng/l, respectively with satisfactory repeatability (RSD 0.31-0.47% for migration time, and 3.0-4.0% for peak height measurement). Agreeable results with established inductively coupled plasma-atomic emission spectrometry method have been obtained for orange juice and tea samples.

Electroencephalogram (EEG) data acquired in the MRI scanner contains significant artifacts, one of the most prominent of which is ballistocardiogram (BCG) artifact. BCG artifacts are generated by movement of EEG electrodes inside the magnetic field due to pulsatile changes in blood flow tied to the cardiac cycle. Independent Component Analysis (ICA) is a statistical algorithm that is useful for removing artifacts that are linearly and independently mixed with signals of interest. Here, we demonstrate and validate the usefulness of ICA in removing BCG artifacts from EEG data acquired in the MRI scanner. In accordance with our hypothesis that BCG artifacts are physiologically independent from EEG, it was found that ICA consistently resulted in five to six independent components representing the BCG artifact. Following removal of these components, a significant reduction in spectral power at frequencies associated with the BCG artifact was observed. We also show that our ICA-based procedures perform significantly better than noise-cancellation methods that rely on estimation and subtraction of averaged artifact waveforms from the recorded EEG. Additionally, the proposed ICA-based method has the advantage that it is useful in situations where ECG reference signals are corrupted or not available.

Recent models of hippocampal function have emphasized its role in processing sequences of events. In this study, we used an oddball task to investigate hippocampal responses to the detection of deviant "target" stimuli that were embedded in a sequence of repetitive "standard" stimuli. Evidence from intracranial event-related potential studies has suggested a critical role for the hippocampus in oddball tasks. However, functional neuroimaging experiments have failed to detect activation in the hippocampus in response to deviant stimuli. Our study aimed to resolve this discrepancy by using a novel functional magnetic resonance imaging (fMRI) technique that drastically improves signal detection in the hippocampus. Significant hippocampal activation was observed during both auditory and visual oddball tasks. Although there was no difference in the overall level of hippocampal activation in the two modalities, significant modality differences in the profile of activation along the long axis of the hippocampus were observed. In both left and right hippocampi, an anterior-to-posterior gradient in the activation (anterior to posterior) was observed during the auditory oddball task, whereas a posterior-to-anterior gradient (posterior to anterior) was observed during the visual oddball task. These results indicate that the hippocampus is involved in the detection of deviant stimuli regardless of stimulus modality, and that there are prominent modality differences along the long axis of the hippocampus. The implications of our findings for understanding hippocampal involvement in processing sequences of events are discussed., (Copyright (c) 2004 Wiley-Liss, Inc.)

Purpose: Various strategies have recently emerged to improve the diagnostic prediction of prostate cancer (CaP). One such strategy includes the mass profiling of serum protein fractions selectively adsorbed onto chemically modified probes. In the current study we further validated this approach, while offering a more versatile, less expensive and yet equally predictive alternative to existing technologies., Materials and Methods: A solid core lipophilic C-18 resin was used to extract and enrich the low molecular weight protein fraction from patient serum for further analysis by mass spectrometry. Mass spectra generated from a 48 patient training set were data mined using multivariate analysis to identify diagnostically significant protein peaks. These peaks were then used to test a blinded study set comprising 168 patients with common statistical algorithms and commercially available software packages., Results: A total of 36 peaks generated from the training set were used to test the combined set of 168 serum samples obtained from 98 healthy individuals and 70 patients with CaP. We report a sensitivity of 94.1% and a specificity of 99.0% with 1 false-positive, 4 false-negative and 5 nondiagnosed cases., Conclusions: Our results further indicate that mass profiling of serological proteins provides a means for the accurate detection of CaP. In addition, our approach was found to be superior to chip based protocols, generating rich, sharp, highly reproducible spectra attainable in a high throughput manner and at minimal cost. This technique is also scaleable for subsequent protein characterization using multidimensional protein identification technologies. Finally, analyses of mass spectra with commercially available statistical applications was found to be highly effective in generating highly discriminatory m/z values for CaP diagnosis.

The methylation status, mutation and expression of RASSF1A, and mutations of RAS and BRAF were studied in 52 patients with multiple myeloma (MM), one plasma cell leukaemia (PCL) patient and four MM-derived cell lines. Aberrant methylation of RASSF1A was found in nine of 32 MM patients and in one of four MM cell lines (U266), where the associated loss of transcription was reversible by demethylation treatment. RASSF1A transcription was further investigated on anti-CD138-sorted plasma cell-enriched bone marrow samples from 10 MM, one PCL and three reactive plasmacytosis patients. While the wild-type RASSF1A transcript was detected in all three reactive plasmacytosis and the PCL samples, we found no detectable wild-type transcripts in six of 10 MM samples studied. In two MM samples, only the non-functional variant transcript was detected, whereas the other four showed loss of transcription. In great contrast to western data, RAS mutations were identified in only four of 31 (13%) MM patients. While no RASSF1A or BRAF mutation (V599E) was detected in any of the primary MM studied (n = 21), the latter was found in the U266 cell line. Taken together, these data indicate that alterations of RAS signalling are critical in MM pathogenesis. In our current studies of Chinese MM patients, these alterations involved frequent RASSF1A inactivation (60%) as a result of transcriptional silencing or expression of a non-functional variant transcript.

A new analytical procedure is developed using a strong complexing agent, 1,10-phenanthroline (Phen), for direct UV detection of Zn, Mn, Cu, Co, Cd, and Fe at microg/L concentrations in environmental water samples. The metal chelates formed showed different electrophoretic mobilities and solved the comigration problem for capillary electrophoresis (CE) separation of free metal ions. To obtain stable metal-Phen chelates during the capillary zone electrophoresis (CZE) run, both pre-column and on-column complexation are required and threefold excess of Phen over metal ions should be added to the sample. The optimized background electrolyte (BGE) consists of 30 mM hydroxylamine hydrochloride and 0.1% methanol at pH 3.6. Under hydrodynamic sampling, CE run at + 20 kV in 65 cm x 0.05 mm ID fused-silica column with detection at 265 nm, baseline separation, satisfactory working ranges (10 microg/L to 5.5 mg/L), sensitive detection limits (1-3 microg/L), good repeatability for migration times (relative standard deviation, RSD 0.36-0.81%, n = 5), peak area (RSD 3.2-4.2%, n = 5) and peak height (RSD 3.2-4.5%, n = 5) were obtained for the metal cations investigated. The reliability of the method was established by parallel determination using the inductively coupled plasma-atomic emission spectrometry (ICP-AES) method giving results within statistical variation. The procedure developed is shown to provide a quick, sensitive, precise, and economic method for simultaneous determination of metal cations that can form stable chelates with Phen.

A new capillary electrophoresis (CE) procedure was developed for simultaneous determination of ten oxoanions (CrO4(2-), SeO4(2-), MoO4(2-), WO4(2-), VO4(3-), SeO3(2-), As04(3-), TeO3(2-), TeO4(2-), and AsO3(3-)) which were baseline-separated from each other and from the interfering UV absorbing anions (NO3- and NO2-) commonly found in environmental water samples. The new background electrolyte system developed contained 5 mM potassium phosphate and 0.007 mM octadecyltrimethylammonium hydroxide, pH 11.2. The optimized working conditions were electrokinetic sampling at -5 kV for 10 s, running voltage at -15 kV with 5 microA current, and detection wavelength at 205 nm. No interference was observed for non-UV-absorbing anions and UV-absorbing anions up to 20 and 10 times higher concentrations respectively. The speed of analysis was fast, with a complete CE run within 6 min. Wide linear ranges (1-2,000 microg/L), good repeatability in migration time (relative standard deviation RSD 0.55-2.8%), satisfactory precision in peak area (RSD 3.8-5.6%) and peak height (RSD 3.9-5.3%) measurement, and detection limits (1-25 microg/L) sufficiently sensitive to detect oxoanions found in environmental water samples were obtained. The reliability of the CE procedure developed had been established by recovery test and parallel method determination using atomic absoprtion spectrophotometry for real river water sample.

An antibody, GC-17, thoroughly characterized for its specificity for estrogen receptor-beta (ER-beta), was used to immunolocalize the receptor in histologically normal prostate, prostatic intraepithelial neoplasia, primary carcinomas, and in metastases to lymph nodes and bone. Comparisons were made between ER-beta, estrogen receptor-alpha (ER-alpha), and androgen receptor (AR) immunostaining in these tissues. Concurrently, transcript expression of the three steroid hormone receptors was studied by reverse transcriptase-polymerase chain reaction analysis on laser capture-microdissected samples of normal prostatic acini, dysplasias, and carcinomas. In Western blot analyses, GC-17 selectively identified a 63-kd protein expressed in normal and malignant prostatic epithelial cells as well as in normal testicular and prostatic tissues. This protein likely represents a posttranslationally modified form of the long-form ER-beta, which has a predicted size of 59 kd based on polypeptide length. In normal prostate, ER-beta immunostaining was predominately localized in the nuclei of basal cells and to a lesser extent stromal cells. ER-alpha staining was only present in stromal cell nuclei. AR immunostaining was variable in basal cells but strongly expressed in nuclei of secretory and stromal cells. Overall, prostatic carcinogenesis was characterized by a loss of ER-beta expression at the protein and transcript levels in high-grade dysplasias, its reappearance in grade 3 cancers, and its diminution/absence in grade 4/5 neoplasms. In contrast, AR was strongly expressed in all grades of dysplasia and carcinoma. Because ER-beta is thought to function as an inhibitor of prostatic growth, androgen action, presumably mediated by functional AR and unopposed by the beta receptor, may have provided a strong stimulus for aberrant cell growth. With the exception of a small subset of dysplasias in the central zone and a few carcinomas, ER-alpha-stained cells were not found in these lesions. The majority of bone and lymph node metastases contained cells that were immunostained for ER-beta. Expression of ER-beta in metastases may have been influenced by the local microenvironment in these tissues. In contrast, ER-alpha-stained cells were absent in bone metastases and rare in lymph nodes metastases. Irrespective of the site, AR-positive cells were found in all metastases. Based on our recent finding of anti-estrogen/ER-beta-mediated growth inhibition of prostate cancer cells in vitro, the presence of ER-beta in metastatic cells may have important implications for the treatment of late-stage disease.

RET fused gene (RFG)/ELE1alpha/androgen receptor-associated protein 70(ARA70) was first found to be involved in the activation of the RET proto-oncogene in thyroid neoplasm and has recently been shown to be a ligand-dependent transcriptional coregulator for androgen receptor (AR). The functionality of RFG/ELE1alpha/ARA70 remains controversial, and little is known about factors regulating its expression in the prostate. Of significant interest is whether this molecule is involved in prostate carcinogenesis. Using reverse transcriptase-polymerase chain reaction semiquantitation, we compared RFG/ELE1alpha/ARA70 mRNA levels in four prostate cancer cell lines (LNCaP, TSU-Pr1, DU-145, and PC-3) with those found in primary cultures of normal prostatic epithelial cells (PrECs). In addition, we examined the effects of androgen and antiandrogen, estrogen and antiestrogen, and a demethylating agent on RFG/ELE1alpha/ARA70 mRNA expression levels in AR- and AR+ PC-3 cells. Reduced levels of RFG/ELE1alpha/ARA70 message were observed in all four prostate cancer cell lines when compared with normal PrECs in primary cultures. RFG/ELE1alpha/ARA70 mRNA levels in PC-3 cells, which express both estrogen receptor subtypes, were upregulated by 17beta-estradiol and inhibited by the antiestrogen ICI-182780. In PC-3(AR+) cells, which were genetically engineered to express AR, exposure to androgen upregulated RFG/ELE1alpha/ARA70 mRNA expression, whereas treatment with 4-hydroxyflutamide lowered expression of this transcript. Furthermore, treatment of DU-145 cells, which did not express RFG/ELE1alpha/ARA70 transcripts, with a demethylating agent reactivated transcription of this gene. Polymerase chain reaction analyses of monochromosomal human-rodent hybrid panels localized a putative RFG/ELE1alpha/ARA70 isoform on human chromosome 5q31.1-31.2. In summary, we identified sex hormones and DNA hypermethylation as regulators of RFG/ELE1alpha/ARA70 expression in prostate cancer cells. In addition, we found reduced levels of RFG/ELE1alpha/ARA70 expression in prostate cancer cell lines when compared with expression levels in normal PrECs in culture. These findings suggest that RFG/ELE1alpha/ARA70 may be involved prostate carcinogenesis and that it may serve as a key mediator of estrogen-androgen synergism.

Germline mutations of BRCA1 and BRCA2 predispose to hereditary breast, ovarian, and possibly prostate cancer, yet structural mutations in these genes are infrequent in sporadic cancer cases. To better define the involvement of these genes in sporadic cancers, we characterized expression levels of BRCA1 and BRCA2 transcripts in cancer cell lines derived from neoplasms of the ovary, prostate, and breast and compared them with those expressed in primary cultures of normal epithelial cells established from these organs. We observed upregulation of BRCA1 and/or BRCA2 expression in six of seven ovarian cancer cell lines (OVCA420, OVCA429, OVCA432, ALST, DOV13, and SKOV3) when compared with levels found in normal ovary surface epithelial cells. Furthermore, five cancerous or immortalized prostatic epithelial cell lines (BPH-1, TSU-Pr1, LNCaP, PC-3, and DU145) also expressed higher levels of BRCA1 and/or BRCA2 mRNA than did primary cultures of normal prostatic epithelial cells. In contrast, only the estrogen receptor-positive MCF-7 cell line overexpressed these messages, whereas the estrogen receptor-negative breast cancer cell lines Hs578T, MDA-MB-231, and MDA-MB-468 showed no change in expression levels when compared with normal breast epithelial cells. In addition, expanding on our recent identification of a novel BRCA2 transcript variant carrying an in-frame exon 12 deletion (BRCA2 delta 12), we report increased expression of this variant in several ovarian, prostate, and mammary cancer cell lines (OVCA420, OVCA433, ALST, DOV13, SKOV3, TSU-Pr1, DU145, and MDA-MB-468). Most notably, high levels of BRCA2 delta 12 mRNA were detected in an estrogen receptor-positive breast cancer cell line, MCF-7, and in an androgen-independent prostate cancer cell line, DU-145. Interestingly, the wild-type BRCA2 transcript was barely detectable in DU145, which could be used as a model system for future investigations on BRCA2 delta 12 function. Taken together, our data suggest disruption of BRCA1 and/or BRCA2 gene expression in certain epithelial cancer cell lines of the ovary, prostate, and breast. Because wild-type BRCA1 and BRCA2 gene products increase during cell-cycle progression and are believed to exert growth-inhibitory action, enhanced expression of these genes in cancer cells may represent a negative feedback mechanism for curbing proliferation in fast-growing cells. At present, the functionality of BRCA2 delta 12 remains elusive., (Copyright 2000 Wiley-Liss, Inc.)

Brandisia hancei is a medicinal herb in China. The ethanol extract of this plant and four phenylethanoid glycosides, acteoside (1), 2'-acetylacteoside (2), poliumoside (3) and brandioside (4), isolated from it were shown to have inhibitory effects on free radical-induced hemolysis of red blood cells and free radical scavenging activities in vitro. Brandioside (4) and poliumoside (3) showed stronger antioxidant effect than acteoside (1), 2'-acetylacteoside (2) and trolox.

The aim of the current study is to demonstrate normal and malignant prostatic epithelial cells (PrECs) as targets for receptor-mediated estrogenic and antiestrogenic action. Using an improved protocol, we have successfully isolated and maintained highly enriched populations of normal PrECs from ultrasound-guided peripheral zone biopsies, individually determined to be morphologically normal. Semiquantitative reverse transcription-PCR analyses were used to determine whether transcripts of estrogen receptor (ER)-alpha and those of ER-beta were expressed in our normal PrEC primary cultures, in a commercially available PrEC preparation (PrEC; Clontech), in an immortalized PrEC line established from a benign prostatic hyperplasia specimen (BPH-1), and in three prostatic cancer cell lines (LNCaP, PC-3, and DU145). Expression levels of ER-alpha and ER-beta transcripts were related to those of two estrogen-responsive genes [progesterone receptor (PR) and pS2], at the message levels, to gain insights into the functionality of the ER subtypes in PrECs. Interestingly, only transcripts of ER-beta, but not those of ER-alpha, were found in our primary cultures of normal PrECs, along with both PR and pS2 mRNA. These data strongly suggest that estrogen action was signaled exclusively via ER-beta in normal human PrECs. In contrast, PrEC (Clontech) and BPH-1 cells expressed both ER-alpha and ER-beta transcripts and no PR nor pS2 mRNA in PrEC and only a minimal level of PR mRNA in BPH-1. Among the three prostate cancer cell lines, LNCaP expressed ER-beta mRNA along with transcripts of PR and pS2, DU145 expressed messages of ER-beta and PR, and PC-3 cells exhibited ER-alpha, ER-beta, and pS2 mRNA. Thus, unlike normal PrECs, expression patterns of these genes in malignant PrECs are more variable. Treatment of prostate cancer cells with demethylation agents effectively reactivated the expression of ER-alpha mRNA in LNCaP and DU145 and that of pS2 message in DU145. These findings provide experimental evidence that ER-alpha gene silencing in prostate cancer cells, and perhaps also in normal PrECs, are caused by DNA hypermethylation. To evaluate the potential of using antiestrogens as prostate cancer therapies, we have assessed the growth-inhibitory action of estrogens (estradiol and diethylstilbestrol) and antiestrogens (4-hydroxy-tamoxifen and ICI-182,780) on PC-3 and DU-145 cells. In PC-3 cells, which express both ER subtypes, estrogens as well as antiestrogens are effective inhibitors. In contrast, in DU145 cells, which express only ER-beta, antiestrogens, but not estrogens, are growth inhibitors. By comparison, ICI 182,780 is the more effective cell growth inhibitor. Importantly, the ICI 182,780-induced antiproliferative effects were reversed by cotreatment of DU145 cells with an ER-beta antisense oligonucleotide, hence lending additional support to a central role played by ER-beta in mediating growth-inhibitory action of antiestrogens.

Background: Exposure of male Wistar rats to estradiol-17beta (E(2)) in the presence or absence of dihydrotestosterone propionate (DHT) was previously shown to result in prostate inflammation. The present study examines, for the first time, changes in the expression level of several proinflammatory genes during the course of this experimentally induced prostatitis., Methods: Adult male Wistar rats were given chronic exposure to E(2) + DHT by capsule implantation or were injected with E(2) for short-term exposure. Semiquantitative RT-PCR was employed to measure changes in proinflammatory transcript levels in the separated lobes of the prostate after various times of exposure to estrogen with or without DHT., Results: We observed an upregulation of IL-1beta, IL-6, MIP-2, and inducible nitric oxide synthase (iNOS) after only 4 days treatment with E(2). After 4 weeks of treatment with E(2) + DHT, a significant increase in transcript levels of IL-4, IL-5, IL-6, MIP-2, eotaxin, and iNOS was detected, while IL-1beta and TNF-alpha transcript levels only increased slightly. No increase in transcript levels for cyclooxygenase-2 (cox-2), IFN-gamma, IL-2, or IL-12 was observed., Conclusions: Upregulation of proinflammatory transcripts occurred shortly after exposure to E(2) and well before any inflammatory cells were observed in the prostate. The pattern of gene expression resembled a T(H)2-type helper-cell response., (Copyright 2000 Wiley-Liss, Inc.)

Two trans-acting hammerhead ribozymes (Rzs), Rz1-2 and Rz4-9, were designed and shown to be effective in cleaving rat or mouse metallothenein (MT)-I and MT-II mRNA, respectively, at position +147/148, in a sequence-specific manner. The catalytic efficiency for Rz1-2 on rat MT-I was found to be 678 M(-1)s(-1) and that for Rz4-9 on rat MT-II cRNA was found to be 372 M(-1)s(-1). An expression vector, pRz(4-9/1-2), containing both Rzs linked in tandem, was constructed and used to transfect NbE-1 cells, an immortalized ventral prostate epithelial cell line. Two stable-transfected lines, NbE-1(Rz2) and NbE-1(Rz3), expressing substantial levels of (Rz1-2/Rz4-9) and minimal levels of MT-I and MT-II transcripts were found to exhibit increased sensitivity to cadmium (Cd)-induced cytotoxicity compared to the parent line. This article is the first to report successful degradation of rodent MT mRNA in vitro and in cellulo by hammerhead Rzs., (Copyright 1999 Academic Press.)

Our understanding of the roles played by sex hormones in ovarian carcinogenesis has been limited by a lack of data concerning the mode of sex hormone action in human ovarian surface epithelial (HOSE) cells, the tissue of origin of >90% of ovarian cancers. We have compared the relative abundance of estrogen receptor (ER)alpha, ERbeta, progesterone receptor (PR), and androgen receptor (AR) mRNA in four primary cultures of HOSE cells obtained from postmenopausal women to those found in late serous adenocarcinoma primary cell cultures and established ovarian cancer cell lines. We observed coexpression of ERalpha and ERbeta mRNA along with AR and PR transcripts in normal HOSE cells and disruption of ERalpha mRNA expression as well as dramatic down-regulation of PR and AR transcript expression in most ovarian cancer cells. In contrast, levels of ERbeta mRNA were unaffected by the malignant state. Additionally, a novel mutation involving a 32-bp deletion in exon 1 of ERalpha transcripts was detected in the SKOV3 cell line. This mutation would explain why SKOV3 was reported to be ER-positive but estrogen-insensitive. Taken together, these findings suggest that estrogens, signaling via either or both ER subtypes, may play an indispensable role in regulating normal HOSE cell functions. Therefore, loss of ERalpha, PR, and AR mRNA expression in HOSE cells may be responsible for neoplastic transformation in this cell type. In contrast, the roles played by ERbeta in normal and malignant HOSE cells remain elusive. Finally, the coexistence of mutated ERalpha mRNA and normal ERbeta transcripts in SKOV3 argues in favor of a dependency of ERbeta action on functional ERalphas.

Highly sensitive, sequence-specific competitive reverse transcriptase-polymerase chain reaction (RT-PCR) protocols were established for the detection and quantification of metallothionein (MT)-I and MT-II messages, in absolute values, in rat tissues. Detection limits for these protocols were in the range of 5 to 10 amol per microgram total RNA. Levels of MT-I and MT-II transcripts in the three major prostatic lobes, kidney, and testis were measured in untreated and cadmium (Cd)-treated rats. The dorsal prostate (DP), lateral prostate (LP), kidney, and testis expressed substantial levels of MT-I and MT-II mRNA while the ventral prostate (VP) had extremely low levels of the transcripts. Cd treatment induced higher levels of MT-I and/or MT-II mRNA expression in all tissues studied with the exception of LP. In the LP, Cd treatment caused reductions of MT-I and MT-II mRNA levels. The Cd-induced levels attained in the VP following Cd exposure were still markedly lower than those found in the kidney, testis, LP, and DP of untreated animals. These findings contradict previous claims that the MT genes in rat VP are unresponsive to Cd activation. The susceptibility of VP to Cd toxicity/carcinogenicity may therefore be explained by low levels of Cd-induced expression rather than lack of induction of MTs., (Copyright 1999 Academic Press.)

Recent molecular cloning of estrogen receptor beta (ERbeta) suggests alternative pathways of estrogen signaling, but little is known concerning the role of ERbeta in the development of human breast cancer. In the present study, expression of ERalpha and ERbeta mRNA was determined in a series of chemically transformed human breast epithelial cells as well as various normal and malignant breast cancer cell lines. We observed a very low level of ERbeta expression in the mortal S130 and the spontaneously immortalized MCF10-F human breast epithelial cell lines. As MCF-10F cells were treated with environmental chemical carcinogens, an elevated level of ERbeta expression was observed in the resultant transformed BP1, D3 and BP1-ras cells. An even higher level of ERbeta expression was detected in the more transformed BP1-E, D3-1 and D3-1-ras cell lines. Therefore, results from our study indicate that expression of ERbeta can be induced in chemical carcinogen-transformed human breast epithelial cells, and the more transformed cells showed higher levels of ERbeta expression, regardless of which chemical carcinogens were initially used for cell transformation. These results suggest that expression of ERbeta may contribute to the initiation and progression of chemical carcinogen-induced neoplastic transformation.

A new analytical procedure was developed for simultaneous determination of ammonium, leachable and total metals in fine and coarse air particulate matters using a new capillary electrophoresis (CE) procedure, with a new buffer system containing 10 mM histidine, 2 mM 18-crown-6 and 8 mM lactic acids with pH adjusted to 4.0. A two complexes system, 18-crown-6 ether and lactic acid, was developed to solve the co-migration problem of NH(4)(+) and K(+) and to give satisfactory separation of transition metals. Satisfactory separation and quantitation of NH(4)(+), K(+), Ca(2+), Na(+) , Mg(2+) and Zn(2+) were obtained using the CE procedure developed for both leachable and total metals in coarse (10-3 mum) and fine (<3 mum) air particulate matters. Wide working ranges (ppb to ppm range) and sensitive detection limits (ppb) were obtained for the cations investigated. The reliability was established by parallel method comparison with the ICP-AES method. The analytical procedure developed is shown to provide a quick, sensitive, precise and economic method for simultaneous determination of ammonium, leachable and total metals in air particulate matters.

A new capillary electrophoresis (CE) procedure was developed for simultaneous determination of both organic and inorganic anions in rain water using a background electrolyte (BGE) containing 5 mM molybdate, 0.15 mM CTAH, 0.01% PVA and 5 mM Tris buffer to adjust pH at 7.9. Under optimised conditions, good repeatability (RSD for sulphate in migration time=0.36% and peak area=4.2%), low detection limit (2 ppb for chloride) and satisfactory working range (50 ppb-20 ppm for hydrodynamic injection, 10 ppb-3 ppm for electrokinetic injection for chloride) were obtained. The reliability of the CE procedure developed was established by satisfactory recovery tests and good agreement of results obtained by both the CE and ion chromatography (IC) methods. The procedure developed had been successfully applied for field monitoring of rainwater showing good repeatability and capability of detecting trace anions at ppb levels beyond the IC working range. Thus, the new CE procedure developed provides a quick, sensitive, economic and reliable method to meet the need for the simultaneous determination of both organic and inorganic anions in the acid rain monitoring programme.

Semiquantitative RT-PCR was used to determine if transcripts of the two estrogen receptor (ER) subtypes, ER alpha and ER beta, and the progesterone receptor (PR) are differentially expressed and/or regulated in the various normal lobes of the Noble (NBL) rat prostate. We found that ER beta mRNA was present at comparable, high levels in all three major prostatic lobes: dorsal (DP), lateral (LP) and ventral (VP) prostate. ER alpha mRNA was, however, expressed at low levels among the various lobes in the following descending order of abundance: LP>DP>VP. Expression of PR transcript was low and paralleled the expression pattern of ER alpha mRNA. Treatments of rats with testosterone (T) plus estradiol-17beta (E2) (T+E2) or T alone induced no discernible alterations in ER alpha, ER beta, and PR mRNA levels in the VP, DP and LP, while those with E2 caused a general decline in the expression of all three transcripts. We then studied the expression of the three receptors in the normal and dysplastic epithelium of the dorsolateral prostates (DLPs) of rats treated with T+E2. Comparable levels of ER beta mRNA were found in microdissected dysplastic and normal epithelia. In contrast, significantly higher levels of PR mRNA were present in epithelial samples from dysplastic acini. ER alpha mRNA was not detected in any of the microdissected epithelial samples. Results from this study suggest that upregulation of PR mRNA expression, likely mediated via ER beta action, is involved in the genesis of T+E2-induced dysplasia in this animal model.

Dietary zinc deficiency in rats induces hyperplasia in the esophagus and increases N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumor incidence. Previous work showed a direct relationship between epithelial cell proliferation and esophageal tumor incidence in rats given multiple doses of NMBA. We investigated the effects of single low doses of NMBA in zinc-deficient rats since a single dose of 5.0 mg/kg was reported to be non-carcinogenic in rats. Zinc-sufficient and deficient rats received a single i.g. dose of NMBA at 0.5 or 2.0 mg/kg. At week 14, tumor incidence was 50% with 0.8 +/- 1.0 tumors/rat, and 80% with 2.2 +/- 1.9 tumors/rat, in deficient groups, D(0.5) and D(2.0), that received the lower and higher dose, respectively. In addition, two small papillomas were found in one out of eight untreated zinc-deficient rats. None of the NMBA-treated or untreated zinc-sufficient rats had any tumors. Esophageal cell proliferation, as determined by proliferating cell nuclear antigen (PCNA) immunohistochemistry, showed that, irrespective of NMBA treatment, deficient esophagi had significant increases in the number of labeled cells, the total number of cells, and the labeling index, as compared with zinc-sufficient ones. Mutations in Ha-ras and p53 genes in esophageal tumors were detected by single strand conformation polymorphism (SSCP) analysis. DNA sequencing of variant conformers revealed a point mutation (GGA-->GAA, codon 12) in Ha-ras in 4/5 (80%) and 5/8 (63%) tumors, from D(0.5) and D(2.0) rats, respectively. Three out of eight tumors from D(2.0) rats exhibited SSCP mobility shifts within p53 exons 5 and 7: two tumors (2/8, 25%) had missense mutations and the third, a silent mutation. Of the two tumors with p53 mutations, one had a double mutation (transition at codon 164, TCA-->TTA; transversion at codon 241, AGT-->TGT), and the other tumor, a transition at codon 172 (AGA-->GGA), with amino acid changes in all cases. In parallel with PCNA expression, elevated p53 expression was associated with hyperplastic and dysplastic regions, as well as with tumors, in deficient esophagi. In short, these data indicate that dietary zinc deficiency, with its associated sustained increased cell proliferation in the esophagus, can drive an otherwise non-tumorigenic dose of NMBA into a highly tumorigenic one.

Abnormalities of chromosome 22q have been well studied as a major molecular genetic event in meningiomas. Chromosome 14q loss has also been shown to be a common phenomenon. However, only a few studies have reported molecular genetic changes of this chromosome in meningioma. In this study, we examined 41 sporadic meningiomas of different histological subtypes and grades with 15 polymorphic microsatellite markers covering a wide region on chromosome 14q. Overall, 37% (15 of 41) of cases showed loss of heterozygosity for one or more allelic markers. Thirty percent (10 of 33) of benign tumors showed allelic losses, whereas 62.5% (5 of 8) of high-grade meningiomas showed loss of heterozygosity. There were altogether eight cases of partial deletions and seven cases of probable monosomy of 14q. Allelic losses of 14q were also commonly seen in recurrent tumors (3 of 3, 100%), parasagittal tumors (4 of 7, 57%), and tumors with transitional subtype (7 of 14, 50%). Among the tumors with allelic losses of 14q, all except one concurrently showed loss of heterozygosity for markers on 22q by our previous study. Of the eight cases with partial deletions, one showed losses on 14q11.1-31, two showed deletions on 14q24.3-31, three showed losses at 14q32.1-32.2, and the remaining two showed deletions at 14q24.3-32.2. We therefore defined two cluster regions of deletion on chromosome 14q: 14q24.3-31 and 14q32.1-32.2. Our studies suggested that more than one tumor suppressor gene(s) residing on distinct regions of chromosome 14q are important in the development and atypical or anaplastic changes in meningiomas.

Detection of human papillomavirus (HPV) was accomplished by utilizing polymerase chain reaction methods on cervical scrapes from a total of 488 Hong Kong Chinese women attending the colposcopy, general, and antenatal clinics. From colposcopy clinic, 10% of normal cervix, 31% of condyloma, 56, 50, and 57% of CIN I, CIN II, and CIN III, respectively, and 88% of squamous cell carcinoma contain HPV. The overall detection rate is 30%. The most prevalent type is 16. Four percent of scrapes from normal cervix of patients attending general gynecology clinic and none from antenatal clinic show HPV. The detection rate appears to be more related to the types of cervical lesions than to the age of the patients.

Monosomy of chromosome 22 or deletions of 22q have been described in meningiomas and astrocytic tumors, the incidence of which is increased in Type 2 neurofibromatosis. Recently, the gene for neurofibromatosis Type 2 (NF2) has been identified at Chromosome 22q12, and a tumor suppression role has been suggested. Because there have been only a few studies of the NF2 gene on central nervous system tumors other than vestibular schwannomas, we investigated the potential role of NF2 as a tumor suppressor gene in a group of sporadic meningiomas and astrocytomas. Forty-four tumors (26 meningiomas and 18 astrocytic tumors of different grades) were screened for NF2 mutations for the entire 17 exons by the polymerase chain reaction-single-strand conformation polymorphism method. In addition, 37 tumors and their respective constitutional deoxyribonucleic acid were analyzed for loss of heterozygosity of 22q alleles by four polymorphic microsatellite markers. Seven inactivating mutations were found in Exons 4, 5, 6, and 10 in 7 of 26 (27%) meningiomas, but none were found in astrocytic tumors. Altogether, 69% of meningiomas and 20% of astrocytic tumors revealed a loss of heterozygosity of 22q markers. All tumors with NF2 mutations showed concurrent loss of alleles on 22q, thus fulfilling Knudson's criteria for tumor suppressor genes in meningiomas. We conclude that inactivation of the NF2 gene is involved in the pathogenesis of a proportion of meningiomas but not in astrocytic tumors. Because many meningiomas and some astrocytic tumors had allelic loss of 22q but intact NF2, there is a possibility that other tumor suppressor genes exist on 22q and may be involved in the pathogenesis of central nervous system tumors.