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Aim: To evaluate whether total splenic artery embolization (TSAE) for patients with hypersplenism delivers better long-term outcomes than partial splenic embolization (PSE)., Methods: Sixty-one patients with hypersplenism eligible for TSAE (n = 27, group A) or PSE (n = 34, group B) were enrolled into the trial, which included clinical and computed tomography follow-up. Data on technical success, length of hospital stay, white blood cell (WBC) and platelet (PLT) counts, splenic volume and complications were collected at 2 wk, 6 mo, and 1, 2, 3, 4 years postoperatively., Results: Both TSAE and PSE were technically successful in all patients. Complications were significantly fewer (P = 0.001), and hospital stay significantly shorter (P = 0.007), in group A than in group B. Post-procedure WBC and PLT counts in group A were significantly higher than those in group B from 6 mo to 4 years (P = 0.001), and post-procedure residual splenic volume in group A was significantly less than that observed in group B at 1, 2, 3 and 4 years post-procedure (P = 0.001). No significant differences were observed in red blood cell counts and liver function parameters between the two groups following the procedure., Conclusion: Our results indicate that TSAE for patients with hypersplenism not only delivers a better long-term outcome, but is also associated with lower complication rates and a shorter hospital stay than PSE.

Objectives: to evaluate the prognostic value of apparent diffusion coefficient (ADC) values from MR diffusion-weighted imaging of unresectable hepatocellular carcinoma after chemoembolization., Methods: our study was proved by our institute and informed consent was obtained from all patients before commencement of the study. Twenty-three patients with unresectable hepatocellular carcinoma were scanned immediately before and after chemoembolization within 24h using conventional anatomical MR imaging and diffusion-weighted imaging, from which ADC values in the lesions were measured. The changes in ADC values after chemoembolization were calculated. The relationship between the lesion ADC and the survival time was analyzed by correlation analysis. The overall cumulative survival was analyzed by the Kaplan-Meier method, and survival curves were compared by the log-rank test., Results: the mean overall survival period was (25.0±8.7) months. The pre-chemoembolization lesion ADC value was (1.36±0.249)×10(-3) mm2/s; the change in ADC values post-chemoembolization was (0.377±0.332)×10(-3) mm2/s. There were significant linear regression relation between the survival time and pre-chemoembolization lesion ADC values (r=-0.698, P<0.001) or the changes in ADC value post-chemoembolization (r=0.702, P<0.001). And Log-rank test showed that pre-chemoembolization ADC values (χ2=7.339, P=0.007) or the changes in ADC value post-chemoembolization (χ2=9.820, P=0.002) significantly influenced the overall cumulative survival., Conclusion: Pre-treatment ADC values as well as changes in ADC values after treatment may provide useful information for predicting survival for patients with unresectable hepatocellular carcinoma., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

The pyrethroid insecticide, cypermethrin has been demonstrated to be an environmental anti-androgen in the androgen receptor (AR) reporter gene assay. The amino- and carboxyl-terminal (N/C) interaction is required for transcription potential of the AR. In order to characterize the anti-androgen effects of cypermethrin involved in the N/C interaction of AR, the mammalian two-hybrid assay has been developed in the study. The fusion vectors pVP16-ARNTD, pM-ARLBD and the pG5CAT Reporter Vector were cotransfected into the CV-1 cells. The assay displayed appropriate response to the potent, classical AR agonist 5α-dihydrotestosterone (DHT) and known AR antagonist nilutamide. The N/C interaction was induced by DHT from 10(-11)M to 10(-5)M in a dose-dependent manner. Nilutamide did not activate N/C interaction, while inhibited DHT-induced AR N/C interaction at the concentrations from 10(-7)M to 10(-5)M. Treatment of CV-1 cells with cypermethrin alone did not activate the reporter CAT. Cypermethrin significantly decreased the DHT-induced reporter CAT expression at the higher concentration of 10(-5)M. The mammalian two-hybrid assay provides a promising tool both for defining mechanism involved in AR N/C interaction of EDCs and for screening of chemicals with androgen agonistic and antagonistic activities. Cypermethrin exhibits inhibitory effects on the DHT-induced AR N/C interaction, while the potency is weaker than that of nilutamide., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Staphylococci are among the most important pathogens causing bloodstream infections associated with implanted medical devices. Control of bacterial adhesion to material surfaces is important for prevention of biofilm formation and biomaterial-associated infections. In this study, we hypothesized that submicron (staphylococcal bacterial dimension) surface textures may reduce the bacterial adhesion via a decrease in surface area that bacteria can contact, and subsequently inhibit biofilm formation. Poly(urethane urea) films were textured with two different sizes of submicron pillars via a two-stage replication process. Adhesion of two bacterial strains (Staphylococcus epidermidis RP62A and S. aureus Newman) was assessed over a shear stress range of 0-13.2 dyn cm(-2) using a rotating disk system in physiological buffer solutions. Significant decreases in bacterial adhesion were observed on textured surfaces for both strains compared with smooth controls. Biofilm formation was further tested on surfaces incubated in solution for either 2 or 5 days and it was found that biofilm formation was dramatically inhibited on textured surfaces. The results of the approaches used in this work demonstrate that patterned surface texturing of biomaterials provides an effective means to reduce staphylococcal adhesion and biofilm formation on biomaterial surfaces, and thus to prevent biomaterial-associated infections., (Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

A utilization study was performed in a 2200-bed tertiary care teaching hospital. Data mining was performed on all nasogastric medication prescriptions for patients hospitalized in 2011. Nurses were interviewed by questionnaire. A PDCA (Plan-Do-Check-Act) cycle was used for continuous quality improvement. The proportion of patients with nasogastric tubes (NGT) was 3.2%. A large number of medical orders (n = 6261) involved nasogastric medications with a package insert particularly noting that they should not be crushed or opened (group 1) or medications without a specific formulation recommendation in the package insert but having evidence discouraging NGT dosing (group 2). Of the nasogastrically administered sustained-release or controlled-release formulations, a sustained-release sodium valproate tablet formulation was the most prescribed drug and a sustained-release 2.5 mg felodipine tablet was prescribed with the highest proportion of NGT dosing [NGT/(NGT + oral) = 12.3%]. Among the nasogastrically administered enteric-coated formulations, a myrtol-standardized enteric-coated capsule formulation was the most prescribed drug and a pantoprazole tablet formulation was prescribed with the highest proportion of NGT dosing [NGT/(NGT + oral) = 19.3%]. Proportions of NGT dosing for amiodarone and carbamazepine (group 2) were 4.8% and 6.3%, respectively. The percentage of nurses with adequate knowledge about pharmaceutical dosage formulations was 60%. The rate of answering correctly as to whether medications in group 1 could be crushed or opened was only 30%. Awareness of evidence discouraging NGT dosing of medications in group 2 was zero. Most nurses (90%) left physicians and pharmacists with the entire responsibility for knowledge and decision-making concerning route of drug administration. After a 3-month preliminary intervention, irrational medical orders involving nasogastric administration of medications in group 1 were successfully abolished. The rate of answering correctly as to whether medications in group 1 could be crushed or opened increased to 100%. This utilization study indicates poor awareness concerning nasogastric administration of medication on the part of physicians and nurses, and preliminary intervention measures were efficient in improving knowledge through team cooperation and effort.

We previously demonstrated that oscillatory fluid flow activates MC3T3-E1 osteoblastic cell calcium signaling pathways via a mechanism involving ATP releases and P2Y(2) puringeric receptors. However, the molecular mechanisms by which fluid flow initiates cellular responses are still unclear. Accumulating evidence suggests that lipid rafts, one of the important membrane structural components, may play an important role in transducing extracellular fluid shear stress to intracellular responses. Due to the limitations of current techniques, there is no direct approach to study the role of lipid rafts in transmitting fluid shear stress. In this study, we targeted two important membrane components associated with lipid rafts, cholesterol, and glycosylphosphatidylinositol-anchored proteins (GPI-anchored proteins), to disrupt the integrity of cell membrane structures. We first demonstrated that membrane cholesterol depletion with the treatment of methyl-β-cyclodextrin inhibits oscillatory fluid flow induced intracellular calcium mobilization and ERK1/2 phosphorylation in MC3T3-E1 osteoblastic cells. Secondly, we used a novel approach to decrease the levels of GPI-anchored proteins on cell membranes by overexpressing glycosylphosphatidylinositol-specific phospholipase D in MC3T3-E1 osteoblastic cells. This resulted in significant inhibition of intracellular calcium mobilization and ERK1/2 phosphorylation in response to oscillatory fluid flow. Finally, we demonstrated that cholesterol depletion inhibited oscillatory fluid flow induced ATP releases, which were responsible for the activation of calcium signaling pathways in MC3T3-E1 osteoblastic cells. Our findings suggest that cholesterol and GPI-anchored proteins, two membrane structural components related to lipid rafts, may play an important role in osteoblastic cell mechanotransduction., (Copyright © 2010 Wiley-Liss, Inc.)

Aim: To study the safety and feasibility of total embolization of the main splenic artery as a supplemental treatment modality for hypersplenism with thrombocytopenia or leukocytopenia accompanying liver cirrhosis., Methods: Fifteen consecutive patients with hypersplenism due to cirrhosis were enrolled in this study from January 2006 to June 2010. All patients underwent total embolization of the main splenic artery. Clinical symptoms, white blood cell (WBC) and platelet (PLT) counts, splenic volume, and complications of the patients were recorded. The patients were followed up for 1 and 6 mo, and 1, 2, 3 years, respectively, after operation., Results: Total embolization of the main splenic artery was technically successful in all patients. Minor complications occurred in 13 patients after the procedure, but no major complications were found. The WBC and PLT counts were significantly higher and the residual splenic volume was significantly lower 1 and 6 mo, and 1, 2, 3 years after the procedure than before the procedure (P < 0.01). Moreover, the residual splenic volume increased very slowly with the time after embolization. All patients were alive during the follow-up period., Conclusion: Total embolization of the main splenic artery is a safe and feasible procedure and may serve as a supplemental treatment modality for hypersplenism with thrombocytopenia or leukocytopenia accompanying liver cirrhosis.

The photoinduced electron transference (PET) interaction in porphyrin containing donor-acceptor (D-A) molecules is of great importance in nature and a significant part of the PET research has been devoted to the study of its mechanism ("through-space" or "through-bond") in these decades. Herein we synthesized a series of covalently linked porphyrin-anthraquinone dyads (Por-C(n)-AQ) bridged with flexible alkoxy chains at different lengths (n=1, 4, 10) and investigated their intramolecular PET using a combination of electronic absorption, steady-state fluorescence and decayed luminescence spectra. The experimental results show that the PET efficiency depends on the length of the flexible linkage between the porphyrin and anthraquinone moieties. Meanwhile, theoretical calculation applying the density functional theory (DFT) was also carried out to give the frontier orbital distribution and the optimized structures of these dyads. It is found that the orientation of the dyad with high PET efficiency is disadvantageous to π-π interaction. Thus, the PET of these dyads seemingly is best compatible with a "through-bond" (superexchange) mechanism., (Copyright © 2010 Elsevier B.V. All rights reserved.)

To study adverse effects and underlying mechanisms of beta-cypermethrin (β-cyp) on male reproductive system, the 15-day intact male adult Sprague-Dawley (SD) rats assay was used as an in vivo test. Male adult SD rats were treated by oral gavage with 0, 15 and 30mgβ-cyp/kgBW for 15 days. After 15-day treatments, the testes, epididymis and seminal vesicles were excised and weighed, respectively. One testis was used for testicular sperm head counts, and the other was for immunohistochemistry test to characterize the expression of androgen receptors (ARs). There were substantial decreases of both sperm head counts and daily sperm production after β-cyp exposure. The expression of AR decreased significantly in rats treated with 15 and 30mgβ-cyp/kgBW, and the gray scale pixel values in the three groups (0, 15 and 30mgβ-cyp/kgBW) were 113.79±13.58, 96.09±5.95 and 77.27±5.44, respectively. These findings suggested β-cyp has significant adverse effects on the reproductive system. Reducing the expression of AR is a potential mechanism of decreased sperm production caused by β-cyp., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Objective: To analyze the clinical features and prognostic factors of primary gastric diffuse large B-cell lymphoma (PG-DLBCL) and to evaluate the staging system and treatment modality of PG-DLBCL., Methods: The clinicopathological data of 69 patients with PG-DLBCL were retrospectively analyzed. Event-free survival (EFS) and overall survival (OS) were the primary endpoints., Results: The EFS rates at 1, 3, and 5 years were 83.8%, 71.1%, and 69.0%, respectively, with a mean EFS of 91.3 months. The 1-, 3-, and 5-year OS rates were 91.3%, 80.3%, and 72.4%, respectively, with a mean OS of 98.8 months. Univariate analysis revealed that either EFS or OS was significantly prolonged by the following factors (P < 0.05): modified Ann Arbor stage I(E) or II(E1) disease; normal lactate dehydrogenase (LDH) level; normal hemoglobin level; normal albumin level; International Prognostic Index (IPI) of 0 or 1; tumor size < 5 cm; and less depth of invasion. While gender, age, B symptoms at presentation, performance status and treatment modality were not significantly associated with the prognosis (P > 0.05). Cox regression model revealed that only modified Ann Arbor stage and albumin level were independent prognostic factors for EFS and OS., Conclusion: The most accurate staging system and the exact role of different therapeutic options for PG-DLBCL are still debated. Further randomized prospective studies with a large number of patients are still needed to establish an optimal management for this disease.

Objective: To investigate the value of hepatocellular carcinoma pretreatment apparent diffusion coefficients (ADCs) and its ADCs changes after treatment in predicting and early monitoring the response after chemoembolization., Materials and Methods: Twenty-five responding and nine nonresponding hepatocellular carcinoma lesions were prospectively evaluated with magnetic resonance diffusion-weighted imaging in 24 h before and in 48 h after chemoembolization. Quantitative ADC maps were calculated with images with b values of 0 and 500 s/mm(2)., Results: Nonresponding lesions had a significantly higher pretreatment mean ADC than did responding lesions (1.726+/-0.323 x 10(-3) mm(2)/s vs.1.294+/-0.18510(-3) mm(2)/s, P< or =0.001). The results of receiver operator characteristic (ROC) analysis for identification of nonresponding lesions showed that threshold ADC value of 1.618 x 10(-3) mm(2)/s had 96.0% sensitivity and 77.8% specificity. After transarterial chemoembolization, responding lesions had a significant increase in %ADC values than did nonresponding lesions (32.63% vs. 5.24%, P=0.025). The results of ROC analysis for identification of responding lesions showed that threshold %ADC value of 16.21% had 72% sensitivity and 100% specificity. No significant change was observed in normal liver parenchyma (P=0.862) and spleen (P=0.052)., Conclusion: High pretreatment mean ADC value of hepatocellular carcinoma was predictive of poor response to chemoembolization. A significant increase in %ADC value was observed in lesions that responded to chemoembolization., (Copyright 2009 Elsevier Ireland Ltd. All rights reserved.)

Microphase separation is a central feature of segmented polyurethane biomaterials and contributes to the biological response to these materials. In this study we utilized atomic force microscopy (AFM) to study the dynamic restructuring of three polyurethanes having soft segment chemistries of interest in biomedical applications. For each of the materials we followed the changes in near surface mechanical properties during hydration, as well as fibrinogen activity and platelet adhesion on these surfaces. Both AFM phase imaging and force mode analysis demonstrated that these polyurethane biomaterials underwent reorientation and rearrangement resulting in a net enrichment of hard domains at the surface. Fibrinogen activity and platelet adhesion on the polyurethane surfaces were found to decrease with increasing hydration time. The findings suggest that water-induced enrichment of hydrophilic hard domains at the surface changes the local surface physical and chemical properties in a way that influences the conformation of fibrinogen, changing the availability of the platelet-binding sites in the protein. This work demonstrates that the hydrated polyurethane biomaterial interface is a complex and dynamic environment where the surface chemistry is changing, altering the activity of fibrinogen and affecting blood platelet adhesion., (Copyright 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Objectives: To investigate the burden of chronic diseases among rural-urban residents in Xuzhou, China., Study Design: Retrospective study., Methods: The mortality rates and standardized mortality rates of neoplasms, diabetes mellitus, ischaemic heart disease and cerebrovascular diseases were calculated for the 10 years from 1997 to 2006 in Xuzhou. The potential years of life lost, rate of life lost and standardized rate of life lost were calculated for each disease. Direct treatment costs were also analysed., Results: The age-standardized mortality rates of the four diseases were 150, 9, 78 and 96 per 100,000, respectively. The standardized rates of life lost among urban residents were 11.7, 0.8, 4.9 and 4.1 per thousand, respectively, and among rural residents were 15.0, 0.3, 2.9 and 2.8 per thousand, respectively. The total direct medical expenses, including outpatient and inpatient costs, was 6.07 hundred million Yuan., Conclusion: Chronic diseases place a heavy burden on rural and urban residents in Xuzhou. A multidimensional and multidisciplinary health promotion and disease management plan is urgently needed to control these diseases., (Copyright 2010 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

We have constructed a novel refractive index (RI) sensor based on a fiber optic Fabry-Perot interferometer (FPI) by splicing a section of hollow core fiber between a single-mode fiber and a photonic crystal fiber (PCF). Owing to the air holes in the cladding of the PCF, various substances, such as liquids and gases with different RI, can enter or leave the in-fiber air cavity, which makes the device usable as a refractometer. In this paper, the fiber optic FPI sensor has been used to monitor the RI changes of air with different pressures, and the experimental results show that such a sensor has an RI sensitivity of 805.1 microm/RIU, and hysteresis is not observed. Moreover, the easy fabrication method gives the in-fiber refractometer many potential applications in the sensing field.

Microphase separation is an important characteristic of polyurethane copolymer biomaterials. An improved understanding of the effects of microphase structure on protein interactions with the polymeric biomaterial surface is essential for the development and application of new biomaterials intended for implantation into the body. In this study, an array of atomic force microscopy (AFM) techniques were used to visualize the phase separation structure in a hydrated poly(urethane urea) (PUU) material and to correlate that structure with molecular interactions at the molecular level. Sequential in situ AFM phase images showed that the hard domains present a dynamic environment and undergo rearrangement and enrichment at the surface when hydrated. Adhesion forces measured using a protein-modified AFM probe suggests that the PUU surface became less adhesive to protein with hydration time, consistent with other physical characterizations. Force measurements were used to quantify and correlate mechanical properties and local adhesion forces for bovine serum albumin, and results showed that low adhesion forces were primarily associated with polar hard domain regions. A nanogold-labeled protein conjugate was used to visualize individual protein adsorption to the separate microstructures on the PUU surface, with preferential protein adsorption seen on the more apolar hydrophobic soft segment regions. Together, the results suggest that the microphase separation structure mediates local surface microenvironments that influence biological interactions with the surface.

Cancer vaccine/immunotherapy rarely involves systemic administration of an immunogenic compound to an actively immunized host. We have developed such a strategy that utilizes folate to deliver antigenic haptens [e.g., fluorescein (FITC) and dinitrophenyl] to folate receptor-positive tumors in a hapten-pre-vaccinated host. Here, we investigated the safety of this novel approach and developed strategies to prevent drug-related hypersensitivity. Using FITC as the model hapten, we identified a potential source of allergic species in folate-FITC preparations by LC-MS/MS. In mice and guinea pigs, we tested the significance of this impurity by passive cutaneous anaphylaxis and active systemic anaphylaxis assays. We studied the effect of immunogen (e.g., KLH-FITC) dose and derived a desensitization regimen that was further evaluated in a murine tumor model. Administration of folate-FITC with low multi-haptenated contaminants (e.g. bis-FITC) resulted in hypersensitivity in underimmunized animals. However, this drug-related hypersensitivity may be independently prevented by (1) increasing the immunogen dose and/or (2) desensitizing animals with folate-FITC during vaccination. In addition, such manipulation in vivo did not appear to negatively alter the effectiveness of immunotherapy. This study provided confidence on the safety of folate-hapten-targeted cancer immunotherapy in an actively immunized host.

Atomic force microscopy (AFM) was used to analyze the interactions between fibrinogen and model surfaces having different levels of water wettability. In contrast to most AFM studies, proteins were coupled to the substrate while model surface colloids were attached to the end of the AFM probe, thereby ensuring that proteins undergo only a single compression/decompression cycle. Similar values of adhesion force were observed between fibrinogen and all of the highly wettable surfaces; in the same manner, fibrinogen showed similar adhesion forces against all poorly wettable surfaces, with a step-like transition observed between the two groups. Relationships between the adhesion forces and loading rates were used to analyze the energy profiles involved in protein/surface interactions. Multiple energy barriers were found in the interaction of proteins with poorly wettable surfaces; whereas a single energy barrier was found for protein interactions with highly wettable surfaces. Contact time-dependent adhesion data were fit to an exponential model and showed that the rate constants of the protein unfolding process on highly wettable surface were smaller at low loading rates, but increased rapidly to yield values similar to those on poorly wettable surfaces at high loading rates. The activation energies of protein unfolding derived from the data offer insight into the role of surface wettability in affecting adhesion, conformational changes, and ultimately, the activity of proteins at biomaterial surfaces.

Objective: To investigate the value of pretreatment and posttreatment changes of apparent diffusion coefficients (ADCs) in predicting response to chemoembolization in liver cancer., Methods: Patients with liver cancer were examined with diffusion-weighted MRI at two b values (0 and 500 s/mm(2)) before and after chemoemblization. Quantitative ADC maps were calculated using images under b values of 0 and 500 s/mm(2). The mean ADC values of lesions before and after chemoemblization were compared. The correlation of response to chemoembolization with ADC value was analyzed., Results: The mean value of pretreatment ADC in non-responding lesions were significantly higher than that in the responding lesions (1.687 x 10(-3) mm(2)/s vs. 1.278 x 10(-3) mm(2)/s, P < 0.05). The results of receiver operator characteristic (ROC) analysis showed that when a threshold ADC value was set on 1.618 x 10(-3) mm(2)/s, the sensitivity and specificity for identification of non-responding lesions were 96.0% and 77.8%, respectively. After transarterial chemoembolization, the responding lesions had a significant increase in ADC values than non-responding lesions (32.6% vs. 5.2%, P = 0.025). The results of ROC analysis indicated that when the changes of ADC value for identification of responding lesions before and after transarterial chemoembolization was > or = 16.2%, the sensitivity and specificity were 72% and 100%, respectively. However, no significant change was observed in normal liver parenchyma and spleen (P > 0.05)., Conclusion: Pretreatment mean ADC value can be used to predict the response to chemoembolization, and for selection of therapy in liver cancer. A significant increase in mean ADC can be observed if the lesions responds to chemoembolization.

The DNA-binding affinities and DNA photocleavage abilities of cationic porphyrin, 5-(4-carboxyphenyl)-10,15,20-tris(4-methylpyridiniumyl)porphyrin (CTMPyP), and its reference compound meso-tetrakis(N-methyl-4-pyridiniumyl)porphyrin (H2TMPyP) have been investigated. The DNA-binding behaviors of the two compounds in NaH2PO4 buffer were compared systematically by using absorption, fluorescence and circular dichroism (CD) spectra, thermal denaturation as well as viscosity measurements. The experimental results show that CTMPyP binds to DNA in an outside binding mode, while H2TMPyP in an intercalative mode. Photocleavage experiments reveal that both two compounds employ 1O2-mediated mechanism in cleaving DNA and H2TMPyP can cleave DNA more efficiently than CTMPyP. Theoretical calculations were carried out with the density functional theory (DFT), and the calculated results indicate that the character and energies of some frontier orbitals of CTMPyP are quite different from those of H2TMPyP. These theoretical results can be used to explain their different DNA-binding modes and affinities to a certain extent.

Four cationic porphyrin-anthraquinone (Por-AQ) hybrids differing in lengths of flexible alkyl linkage, 5-[4-(1-N-anthraquinonon-yl)-L-oxophenyl]-10,15,20-tris(N-methylpyridinium-4-yl)porphyrin triiodide, (L = acetyl, pentanoyl, octanoyl, undecanoyl, designed as [AQATMPyP]I3, [AQPTMPyP]I3, [AQOTMPyP]I3 and [AQUTMPyP]I3, respectively, see Fig. 1), were synthesized and their interactions with DNA were investigated. The results of spectroscopic, denaturation and viscosity measurements suggest that [AQATMPyP]I3 binds to DNA through non-intercalative mode while the other three hybrids with longer links bind via bis-intercalative mode. Ethidium bromide (EB) competition experiment was carried out to determine the binding constants (Kb) of these compounds for CT DNA, and [AQPTMPyP]I3 shows the largest Kb among these hybrids. The photocleavage mechanism and wavelength-dependent cleaving abilities of these hybrids to pBR322 plasmid DNA were also comparably investigated.

The thermodynamics of the binding of a series of structurally related Ru(II) antitumor complexes, that is, alpha-[Ru(azpy)2Cl2] 1, beta-[Ru(azpy)2Cl2] 2, alpha-[Ru(azpy)(bpy)Cl2] 3, and cis-[Ru(bpy)2Cl2] 4 to DNA purine bases (gunine, adenine at N7 site) has been studied by using the DFT method. The binding of imine form of 9-methyladenine (9-MeAde) to the Ru(II) moiety in a didentate fashion via its N6 and N7 atoms was also considered. The geometrical structures of the DNA model base adducts were obtained at the B3LYP/(LanL2DZ + 6-31G(d)) level in vacuo. The following exact single-point energy calculations were performed at the B3LYP/(LanL2DZ(f)+6-311+G(2d, 2p)) level both in vacuo and in aqueous solution using the COSMO model. The bond dissociation enthalpies and free energies, reaction enthalpies and free energies both in the gas phase and in aqueous solution for all considered Ru(II)-DNA model base adducts were obtained from the computations. The calculated bond dissociation enthalpies and free energies allow us to build a binding affinity order for the considered Ru(II)-DNA model base adducts. The theoretical results show that the guanine N7 is a preferred site for this series of complexes and support such an experimental fact that alpha-[Ru(azpy)(bpy)(9-EtGua)H2O](2+) (3-(9-EtGua)) is isomerized to alpha'-[Ru(azpy)(bpy)(9-EtGua)H2O](2+) (3'-(9-EtGua)). On the basis of structural and thermodynamical characteristics, the possible structure-activity relationship was obtained, and the distinct difference in cytotoxicities of this series of structurally related antitumor complexes was explained theoretically.

Four tricationic pyridium porphyrins appending hydroxyphenyl, methoxyphenyl, propionoxyphenyl or carboxyphenyl group at meso-20-position of porphyrin core have been synthesized and their abilities to bind and cleave DNA have been investigated. Using a combination of absorption, fluorescence, circular dichroism (CD) spectra, thermal DNA denaturation as well as viscosity measurements, their binding modes and intrinsic binding constants (K(b)) to calf DNA (CT DNA) were comparatively studied and also compared with those of 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin (TMPyP). The results suggest that the K(b) values of these porphyrins are greatly influenced by the number of positive charges and steric hindrance. Theoretical calculations applying the density functional theory (DFT) have been carried out and explain their DNA-binding properties reasonably. The efficiency of DNA photocleavage by these porphyrins shows high dependence on the values of K(b).

A novel cationic porphyrin-anthraquinone (Por-AQ) hybrid has been synthesized and characterized. Using the combination of absorption titration, fluorescence spectra, circular dichroism (CD) as well as viscosity measurements, the binding properties of the hybrid to calf thymus (CT) DNA have been investigated compared with its parent porphyrin. The experimental results show that at low [Por]/[DNA] ratios, the parent porphyrin binds to DNA in an intercalative mode while the hybrid binds in a combined mode of outside binding (for porphyrin moiety) and partial intercalation (for anthraquinone). Ethidium bromide (EB) competition experiment determined the binding affinity constants (K(app)) of the compounds for CT DNA. Theoretical calculational results applying the density functional theory (DFT) can explain the different DNA binding behaviors reasonably. (1)O(2) was suggested to be the reactive species responsible for the DNA photocleavage of porphyrin moieties in both two compounds. The wavelength-depending cleavage activities of the compounds were also investigated.

Alkylphenols (APs) are widely used as important industrial materials and have attracted lots of attention because of their potential estrogenic activities. In this study, we developed human estrogen receptor alpha (hERalpha) and rat estrogen receptor alpha (rERalpha) mediated reporter gene assays and compared the estrogenic activity of APs and related chemicals based on the two ERalpha. Human breast cancer cell line MCF-7 was co-transfected with Gal4-fused hERalpha and corresponding reporter plasmid; African green monkey kidney cell line CV-1 was co-transfected with rERalpha and reporter gene. Both assays showed acceptable response to natural estrogen 17beta-estradiol (E2) with EC50 of 0.16 nM and 4.7 nM. Then the estrogenic activity of 4-APs, 4-phenylphenol and bisphenol-A were evaluated and compared with the effects of E2. The data suggested that test APs and related chemicals possessed weakly estrogenic activity and the activity of test APs increased with the increase of substituent size. This structure-activity relationship helped to infer the activity of chemicals with similar feature. Furthermore, test APs showed similar effect on the function of hERalpha and rERalpha. This consistency helped to extrapolate in vivo rodent data to human being when performing risk assessment of endocrine disruptors.

A combined computational and experimental study on DNA-photocleavage by Ru(II) polypyridyl complexes [Ru(bpy)2(L)]2+ 1-3 (bpy = 2,2-bipyridine; L: pip = 2-phenylimidazo[4,5-f]1,10-phenanthroline, o-mopip = 2-(2-methoxyphenyl)imidazo[4,5-f]1,10-phenanthroline and p-mopip = 2-(4-methoxyphenyl)imidazo[4,5-f]1,10-phenanthroline) has been carried out. The DNA-photocleavage behavior of these complexes was comparably measured by the gel electrophoresis experiments. The experimental results show that they can induce considerable DNA-photocleavage, and have different DNA-photocleavage efficiencies (phi) following the order phi (1) < phi (2) < phi (3). In order to understand their DNA-photocleavage mechanism and trend, the theoretical studies on the geometric and electronic structures of these complexes in the ground state (S0), the first singlet excited state (S1) and triplet excited states (T1), have been carried out using the density functional theory (DFT/TD-DFT), Hartree-Fock (HF) and configuration interaction singles (CIS) methods. In particular, the reduction potentials (E*red) of the excited complexes in aqueous solution, which seem to be closely responsible for the DNA-photocleavage behavior, were calculated to be 0.966 V (vs. SCE) for complex , 1.024 V (vs. SCE) for complex and 1.030 V (vs. SCE) for complex , respectively. Such computational results show that the reduction potentials of the excited complexes reach the theoretical range for oxidizing some DNA-bases, and follow the order E*red (1) < E*red (2) < E*red (3). Therefore, here, in addition to the general theoretical explanation of their DNA-photocleavage mechanism according to our recent report, a further explanation on the trend of their DNA-photocleavage efficiencies, i.e., phi (1) < phi (2) < phi (3), was reasonably carried out, on the basis of the calculated electrochemical properties in the excited states as well as general photochemical insights.

Organophosphates and pyrethroids are among the most common pesticides currently in use worldwide. Several pesticides have been reported to possess hormonal activities, and thus are classified as endocrine disruptors. The present study was planned to evaluate potential androgenic and antiandrogenic activities of the pesticides. The selected chemicals are three organophosphate pesticides including dichlorvos, parathion and trichlorphon, and two pyrethroid pesticides including permethrin and cypermethrin. We evaluated the pesticides for androgen receptor (AR)-mediated mechanisms using a human AR reporter gene assay in African monkey kidney cell line CV-1 transiently transfected with the constructed reporter gene plasmid pMMTV-CAT and the hAR expression plasmid AR/pcDNA3.1. We demonstrated that parathion showed significant inhibitory effects on the transcriptional activity induced by 1 nM of DHT with IC(50) value of (2.01+/-0.42) x 10(-7) M, though trichlorphon and dichlorvos lacked this activity. The two pyrethroid pesticides permethrin and cypermethrin exhibited lower activity than parathion with IC(50) value of (5.68+/-2.20) x 10(-5) and (6.80+/-2.30) x 10(-5) M, respectively. On the other hand, we failed to find AR-mediated androgenic activities of the tested chemicals. It is suggested that parathion possesses the highest activity, and permethrin and cypermethrin acted as poor antiandrogens. The present study provides insight into the mechanism of the pesticides.

Atomic force microscopy (AFM) was used to directly measure the adhesion forces between three test proteins and low density polyethylene (LDPE) surfaces treated by glow discharge plasma to yield various levels of water wettability. The adhesion of proteins to the LDPE substrates showed a step dependence on the wettability of surfaces as measured by the water contact angle (theta). For LDPE surfaces with theta> approximately 60-65 degrees , stronger adhesion forces were observed for bovine serum albumin, fibrinogen and human FXII than for the surfaces with theta<60 degrees . Smaller adhesion forces were observed for FXII than for the other two proteins on all surfaces although trends were identical. Increasing the contact time from 0 to 50s for each protein-surface combination increased the adhesion force regardless of surface wettability. Time varying adhesion data was fit to an exponential model and free energies of protein unfolding were calculated. This data, viewed in light of previously published studies, suggests a 2-step model of protein denaturation, an early stage on the order of seconds to minutes where the outer surface of the protein interacts with the substrate and a second stage involving movement of hydrophobic amino acids from the protein core to the protein/surface interface. Impact statement: The work described in this manuscript shows a stark transition between protein adherent and protein non-adherent materials in the range of water contact angles 60-65 degrees , consistent with known changes in protein adsorption and activity. Time-dependent changes in adhesion force were used to calculate unfolding energies relating to protein-surface interactions. This analysis provides justification for a 2-step model of protein denaturation on surfaces.

A total of 180 farmland soil samples were collected from the suburbs of Fuxin City, Liaoning Province, to investigate the contents and spatial distribution patterns of Cu, Zn, As and Ni, with the effects of mining on their accumulation and spatial distribution in test farmland soils discussed. The results indicated that in study area, farmland soils had an accumulation of Cu, Zn and Ni, with the geometric mean concentrations higher than the background values. Among the soils planted with different crops, vegetable soil had significantly higher contents of Cu, Zn and As than grain crop soil. The spatial distribution of four test metals showed the same pattern of urban > suburban, and the regions with high concentrations of heavy metals were well overlapped with mining areas, suggesting that mining activities might be one of the most important source of heavy metals in the farmland soils. The rapid increase of animal manure also contributed to the import of pollutants to farmland soils to some degree. The potential risks of soil heavy metals pollution should be considered in the arrangement of local agricultural and animal husbandry productions.

We recently developed a new group of folate-conjugated Vinca alkaloids, one of which, EC145, emerged as a candidate for clinical development. Brief treatment of nude mice bearing approximately 100 mm(3) folate receptor-positive human xenografts led to complete response (CR) in 5/5 mice and cures (i.e., remission without a relapse for >90 days post-tumor implantation) in 4/5 mice. Multiple CRs and cures were also noted when EC145 was used to treat mice initially bearing tumors as large as 750 mm(3). Likewise, complete cures (5/5) resulted following the treatment of an aggressive folate receptor-positive J6456 lymphoma model. The activity of EC145 was not accompanied by noticeable weight loss or major organ tissue degeneration. Furthermore, no significant antitumor activity (0/5 CR) was observed in EC145-treated animals that were co-dosed with an excess of a benign folate ligand, thus demonstrating the target-specific activity of EC145. The enhanced therapeutic index due to folate conjugation was also evidenced by the fact that the unconjugated drug (desacetylvinblastine monohydrazide) was found to be completely inactive when administered at nontoxic dose levels and only marginally active when given at highly toxic dose levels. Subsequent dose regimen studies confirmed that EC145 given on a more frequent, qdx5 schedule resulted in the most effective antitumor response as compared with an equivalent total dose given on thrice- or single-injection-per-week schedule. Taken together, these studies show that EC145 has significant antiproliferative activity and tolerability, thus lending support to an ongoing phase 1 trial for the treatment of advanced malignancies.

The structures and related properties of the complex [Ru(phen)2(6-OH-dppz)]2+ (phen = 1,10-phenanthroline; dppz = dipyrido [3,2-a:2',3'-c]phenazine) in the ground state (S0), the first singlet excited state (S1), and the first triplet excited state (T1) have been studied using density functional theory (DFT), time-dependent (TD) DFT, Hartree-Fock (HF), and configuration interaction singles (CIS) methods. Three electronic absorption-spectral bands (1MLCT, 1LL, and 1LL) lying in the range of 250-550 nm in vacuo and in aqueous solution were theoretically calculated, simulated, and assigned with TDDFT method. In particular, the theoretical results show the following: (1) The positive charges of central Ru atom in the excited states (S1 and T1) are greatly increased relative to those in the ground state (S0), and thus the Ru atom in the excited states can be regarded as Ru(III). (2) The positive charges on the main ligand (6-OH-dppz) in the excited states are considerably reduced, and thus the interaction between the main ligand (intercalative ligand) and DNA base pairs is considerably weakened. (3) The geometric structures in excited states are also distorted, resulting in obvious increase in the coordination bond length. It is advantageous to the complex forming a high oxidizing center (i.e., Ru(III) ion). On the basis of these results, a theoretical explanation on photoinduced oxidation reduction mechanism of DNA photocleavage by [Ru(phen)2(6-OH-dppz)](2+) has been presented.

Atomic force microscopy utilizing both tapping mode and force mode imaging is used to visualize the separated microphases in poly(urethane urea) films under ambient and aqueous conditions. The topography of the PUU surface changed upon hydration with the formation of nanometer-sized features on the surface. The surface becomes enriched in hard domains with hydration time and this enrichment is irreversible after dehydration. Force mode measurements were used to quantify mechanical properties as both indentation and modulus measurements. Analysis of the modulus during indentation reveals the three-dimensional nature of the structures, with the surface being covered by a 2-20-nm-thick soft segment overlayer under ambient conditions, while hydration leads to the loss of this overlayer. The force measurements also reveal the presence of regions having modulus values between those of the hard and soft phases and located spatially near the interface between the hard and soft domains. However, such regions with intermediate modulus were only rarely seen following hydration. Calculation of the Young's modulus from the compression data shows that hydration increases the modulus of the PUU surface by both enrichment of the amount of hard domain present and increasing the modulus of the individual hard and soft phases themselves. Direct visualization of the distribution of these different domains on the surface by nanoscale measurements provides an important path to characterizing the relationships between the surface properties of these materials and subsequent performance in biomedical applications.

Massive amounts of pig manure are produced by intensive pig farm in China, and the composition of pig manure has changed much due to the use of feed additives. However, little is known about the exact Cu (copper) feed as additives or present as contaminants in pig feed and the residues in feces. One hundred and thirty-seven feeds and one hundred and forty-two fecal samples from 48 pig farms were collected in Beijing and Fuxin cities in 1999 and 2005, respectively. The concentrations of Cu were in the range of 6.86-395.19 mg/kg in the feed samples, and the mean values were in the order of weaner > grower-finisher > sow's feeds. The high concentrations over EU recommendations implied that excessive levels of Cu are fed on many pig farms in Beijing and Fuxin. Cu was also present in high concentrations in feces, and concentrations were highly variable. Cu concentrations in the feces from grower-finisher and weaner pigs were significantly greater than feces of sows. The super-intensive and small-scale farms had higher levels of Cu in feces than the middle farms. Cu concentrations in pig feces were approximately 5-times greater than in pig feeds. Feed management in grower-finisher pigs on super-intensive and small-scale pig farms is needed to reduce high Cu concentrations in feces and risks to soil contamination while feces are land-applied.

Mechanical stimulation of osteoblasts by fluid flow promotes a variety of pro-differentiation effects and improving the efficiency of these mechanical signals could encourage specific differentiation pathways. One way this could be accomplished is by altering mechanical properties of osteoblasts. In this study, murine osteoblastic MC3T3-E1 cells were cultured on surfaces covered with nanometer-sized islands to examine the hypothesis that the elastic modulus of osteoblastic cells is affected by nanoscale topography. Nanoislands were produced by polymer demixing of polystyrene and poly(bromostyrene), which leads to a segregated polymer system and formation of nanometer-sized topographical features. The elastic modulus of MC3T3-E1 cells was determined using atomic force microscopy in conjunction with the Hertz mathematical model. Osteoblastic cells cultured on nanotopographic surfaces (11-38 nm high islands) had a different distribution of cellular modulus values, e.g., the distribution shifted toward higher modulus values, relative to cells on flat control surfaces. There were also differences in cell modulus distribution between two flat controls as surface chemistry was changed between polystyrene and glass. Taken together, our results demonstrate that both surface nanotopography and chemistry affect the mechanical properties of cells and may provide new methods for altering the response of cells to external mechanical signals.

Many pesticides possess hormonal activity and have thus been classified as endocrine disruptors. Pyrethroids are commonly used pesticides worldwide, but little has been done to characterize their antiandrogenic activity potential. We tested three frequently encountered pyrethroids (fenvalerate, cypermethrin, permethrin) and their metabolite 3-phenoxybenzoic acid (3-PBA) for antiandrogenic and androgenic activity using a human androgen receptor (AR) mediated luciferase reporter gene assay in CV-1 African green monkey kidney cell. The assay displayed appropriate response to the known AR agonist 5alpha-dihydrotestosterone and AR antagonist nilutamide and flutamide. At 0.1mM, all the three tested pyrethroids significantly suppressed the luciferase expression. Further, their metabolite 3-PBA also showed antagonist activity. None of the test chemicals showed androgenic activity. Through the antiandrogenic pathways, exposure to certain pyrethroids may contribute to the damage of reproductive system. In conclusion, pyrethroid pesticides can act as antiandrogen in vitro, and metabolizing to 3-PBA cannot eliminate the antagonist activity. This result provides useful information for risk assessment of pyrethroid pesticides.

Folic acid (pteroylglutamic acid) represents a useful ligand for targeted cancer therapies because it binds to a common epithelial tumor antigen known as the folate receptor. We previously devised an immunotherapy strategy that uses a bispecific ligand, a folate-hapten (FITC) conjugate, to redirect endogenously induced anti-FITC antibodies to folate receptor-positive tumor cells following parenteral administration. Here, we present results from preclinical pharmacokinetic and tissue biodistribution studies using a radioactive folate-FITC conjugate and results from dose optimization studies done in tumor-bearing animals. Folate-FITC was found to be rapidly eliminated in non-immunized mice; however, in immunized hosts, folate-FITC was shown to form immune complexes with FITC-specific antibodies, the consequence of which was a approximately 173-fold increase in drug exposure (i.e., area under the curve). Using a newly developed ELISA assay, the extent of circulating anti-FITC antibodies occupied by parenterally given folate-FITC was determined to be proportional to the given dose. Furthermore, high doses of folate-FITC were found to promote the cosaturation of tumor cell surface folate receptors and circulating FITC-specific antibodies, blocking the immune recognition of tumor cells and thereby reducing antitumor activity. Nonetheless, by extending the duration of treatment and administering subsaturating doses of folate-FITC, enhanced antitumor response was observed in mice bearing established folate receptor-positive M109 tumors. Overall, results from the present study may help to guide clinicians through on-going clinical investigations of folate-targeted immunotherapy.

Previously, we developed an androgen receptor (AR)-mediated reporter gene assay system, and verified the antiandrogenic activity of bisphenol A (BPA) based on this assay. There were some similar phenol-ring structure chemicals which were widely used while little had been done to evaluate their effect to androgen. In the present study, we applied this assay to evaluate the androgenic and antiandrogenic activities of some phenolic chemicals including BPA, tetrachlorobisphenol A (TCBPA), and pentachlorophenol (PCP). The results showed that BPA exhibited potent antiandrogenic activity with IC50 value of 2.14 microM. TCBPA showed relative weaker activity with IC50 value of 10.45 microM and PCP did not show AR antagonistic activity. On the other hand, we failed to find AR agonistic effects of the three chemicals. To study the reason why BPA and TCBPA reduced the reporter gene expression, we increased the DHT concentration to 100 nM and 10 microM in the reporter assay. Then, we found that the antagonist activity of BPA and TCBPA decreased with the increase of concentration of DHT. This result indicated that these two test chemicals could compete with DHT for the binding of AR. When comparing the structure difference between BPA, TCBPA and PCP, we got a preliminary founding that 3,5-substituents of phenol ring of BPA decrease its antiandrogenic activity. These results provide some useful information for the assessment of the antiandrogenic activity of BPA like chemicals and the potential mechanism.

Studies on the electronic structures and trend in DNA-binding affinities of a series of Ru(II) complexes [Ru(bpy)2(p-R-pip)]2+ (bpy = 2,2-bipyridine; pip = 2-phenylimidazo[4,5-f] [1,10]-phenanthroline; R = -OH, -CH3, -H, -NO2) 1-4 have been carried out, using the density functional theory (DFT) at the B3LYP/LanL2DZ level. The electronic absorption spectra of these complexes were also investigated using time-dependent DFT (TDDFT) at the B3LYP//LanL2DZ/6-31G level. The computational results show that the substituents on the parent ligand (pip) have a significant effect on the electronic structures of the complexes, in particular, on the energies of the lowest unoccupied molecular orbital (LUMO) and near some unoccupied molecular orbitals (LUMO+x, x = 1-4). With the increase in electron-withdrawing ability of the substituent in this series, the LUMO+x (x = 0-4) energies of the complexes are substantially reduced in order, for example, epsilon(LUMO)(1) approximately epsilon(LUMO)(2) > epsilon(LUMO)(3) > epsilon(LUMO)(4), whereas the pi-component populations of the LUMO+x (x = 0-4) are not substantially different. Combining the consideration of the bigger steric hindrance of complex 2, the trend in DNA-binding affinities (K(b)) of the complexes, that is, K(b)(2) < K(b)(1) < K(b)(3) < K(b)(4) can be reasonably explained. In addition, the experimental singlet metal-to-ligand charge transfer ((1)MLCT) spectra of these complexes can be well simulated and discussed by the TDDFT calculations.

Both proteins and polysaccharides are biopolymers present on a bacterial surface that can simultaneously affect bacterial adhesion. To better understand how the combined presence of proteins and polysaccharides might influence bacterial attachment, adhesion forces were examined using atomic force microscopy (AFM) between colloids (COOH- or protein-coated) and polymer-coated surfaces (BSA, lysozyme, dextran, BSA+dextran and lysozyme+dextran) as a function of residence time and ionic strength. Protein and dextran were competitively covalently bonded onto glass surfaces, forming a coating that was 22-33% protein and 68-77% dextran. Topographic and phase images of polymer-coated surfaces obtained with tapping mode AFM indicated that proteins at short residence times (<1 s) were shielded by dextran. Adhesion forces measured between colloid and polymer-coated surfaces at short residence times increased in the order protein+dextran < or = protein < dextran. However, the adhesion forces for protein+dextran-coated surface substantially increased with longer residence times, producing the largest adhesion forces between polymer coated surfaces and the colloid over the longest residence times (50-100 s). It was speculated that with longer interaction times the proteins extended out from beneath the dextran and interacted with the colloid, leading to a molecular rearrangement that increased the overall adhesion force. These results show the importance of examining the effect of the combined adhesion force with two different types of biopolymers present and how the time of interaction affects the magnitude of the force obtained with two-polymer-coated surfaces.

Proteins are important in bacterial adhesion, but interactions at molecular-scales between proteins and specific functional groups are not well understood. The adhesion forces between four proteins [bovine serum albumin (BSA), protein A, lysozyme, and poly-d-lysine] and COOH, NH2 and OH-functionalized (latex) colloids were examined using colloid probe atomic force microscopy (AFM) as the function of colloid residence time (T) and solution ionic strength (IS). For three of the proteins, OH-functionalized colloids produced higher adhesion forces to proteins (2.6-30.5 nN; IS=1 mM, T=10s) than COOH- and NH2-functionalized colloids (1.6-6.8 nN). However, protein A produced the largest adhesion force (8.1+/-1.0 nN, T=10 s) with the COOH-functionalized colloid, demonstrating the importance of specific and unanticipated protein-functional group interactions. The NH2-functionalized colloid typically produced the lowest adhesion forces with all proteins, likely due to repulsive electrostatic forces and weak bonds for NH2-NH2 interactions. The adhesion force (F) between functionalized colloids and proteins consistently increased with residence time (T), and data was well fitted by F=ATn. The constant value of n=0.21+/-0.07 for all combinations of proteins and functionalized colloids indicated that water exclusion and protein rearrangement were the primary factors affecting adhesion over time. Adhesion forces decreased inversely with IS for all functional groups interacting with surface proteins, consistent with previous findings. These results demonstrate the importance of specific molecular-scale interactions between functional groups and proteins that will help us to better understand factors colloidal adhesion to surfaces.

In this study, we have developed a transient human androgen receptor (hAR) reporter gene assay using African monkey kidney cell line CV-1. The assay displayed appropriate response to the known androgen receptor (AR) agonist 5alpha-dihydrotestosterone (DHT) and AR antagonist nilutamide. DHT induced AR-mediated transcriptional activity in a concentration-dependent manner with median effective concentration (EC(50)) value of 3.90 x 10(-10) M. Nilutamide exhibited potent antiandrogenic activity with median inhibitory concentration (IC(50)) value of 8.90 x 10(-7) M. The transcription could not be activated by glucocorticoid receptor (GR) agonist dexamethasone, which suggested that the assay system be highly specific to androgenic compounds without cross-talk to GR agonist. The assay shows acceptable repeatability to DHT with intra coefficient of variability (CV) of 9.3% and inter CV of 10.9%. We applied this assay to evaluate the androgenic and antiandrogenic activities of some pesticides including organochlorine p,p'-dichlorodiphenyldichloroethylene (DDE), pyrethroid fenvalerate and organophosphorus phoxim. The results showed that p,p'-DDE exhibited potent antiandrogenic activity. Fenvalerate acted as weak AR antagonist and phoxim did not show AR antagonistic activity. We failed to find AR agonistic effects of the three pesticides. The present study provided a promising tool to screen for AR agonists and antagonists.

In this study, we have developed a human androgen receptor (hAR) reporter gene assay using African monkey kidney cell line CV-1 transiently transfected with the constructed reporter gene plasmid pMMTV-CAT and the hAR expression plasmid AR/pcDNA3.1. The assay displayed appropriate response to the known AR agonist 5alpha-dihydrotestosterone (DHT) and AR antagonist flutamide. DHT induced AR-mediated transcriptional activity in a concentration-dependent manner with median effective concentration (EC50) value of (3.90+/-1.43)x10(-10)M. Flutamide exhibited potent antiandrogenic activity with median inhibitory concentration (IC50) value of (1.02+/-0.35)x10(-7)M. Bisphenol A (BPA) and alkylphenols (APs) belong to the industrial chemicals that have received considerable attention due to high production and widespread usage. We investigated BPA, 4-octylphenol (OP) and 4-nonylphenol (NP) for their agonistic and antagonistic activities by the AR reporter gene assay. BPA showed significant inhibitory effects on the transcriptional activity induced by DHT with IC50 value of (7.46+/-1.23)x10(-7)M. OP and NP exhibited lower antiandrogenic activities than BPA with IC(50) of (9.71+/-3.82)x10(-5)M and (2.02+/-0.90)x10(-5)M, respectively. On the other hand, we failed to find AR-mediated androgenic activities of the three tested chemicals. In conclusion, BPA, OP and NP all act as AR antagonists, and BPA possesses the highest activity.

The vibrational spectra of meso-tetraphenylporphyrin diacid (H4TPP2+) have been studied with the density functional theory. Raman and IR spectra of H4TPP2+ and its N-deuterated analogue (D4TPP2+) are measured and compared with the computational results. Complete assignments of observed IR and Raman bands were proposed on the bases of calculation results. The DFT calculations reproduce 140 observed fundamentals with the RMS 8.6 cm-1. The computational as well as the experimental results reveal that the saddle-distortion of porphyrin macrocycle for the diacid leads to a significant effect on its vibrational spectra. Especially, several out-of-plane skeletal modes, which were either unobserved or very weak in the Raman spectra of CuTPP and H2TPP, are activated in the Raman spectra of the diacids. In addition, enhancement for the Raman bands of phenyl CC stretching modes were observed and attributed to the conjugation effect of pi-systems of the phenyl and the porphyrinato macrocycles.

Exopolymers are thought to influence bacterial adhesion to surfaces, but the time-dependent nature of molecular-scale interactions of biopolymers with a surface are poorly understood. In this study, the adhesion forces between two proteins and a polysaccharide [Bovine serum albumin (BSA), lysozyme, or dextran] and colloids (uncoated or BSA-coated carboxylated latex microspheres) were analyzed using colloid probe atomic force microscopy (AFM). Increasing the residence time of an uncoated or BSA-coated microsphere on a surface consistently increased the adhesion force measured during retraction of the colloid from the surface, demonstrating the important contribution of polymer rearrangement to increased adhesion force. Increasing the force applied on the colloid (loading force) also increased the adhesion force. For example, at a lower loading force of approximately 0.6 nN there was little adhesion (less than -0.47 nN) measured between a microsphere and the BSA surface for an exposure time up to 10 s. Increasing the loading force to 5.4 nN increased the adhesion force to -4.1 nN for an uncoated microsphere to a BSA surface and to as much as -7.5 nN for a BSA-coated microsphere to a BSA-coated glass surface for a residence time of 10 s. Adhesion forces between colloids and biopolymer surfaces decreased inversely with pH over a pH range of 4.5-10.6, suggesting that hydrogen bonding and a reduction of electrostatic repulsion were dominant mechanisms of adhesion in lower pH solutions. Larger adhesion forces were observed at low (1 mM) versus high ionic strength (100 mM), consistent with previous AFM findings. These results show the importance of polymers for colloid adhesion to surfaces by demonstrating that adhesion forces increase with applied force and detention time, and that changes in the adhesion forces reflect changes in solution chemistry.

A novel folate conjugate of mitomycin C, herein referred to as EC72, was designed and evaluated for biological activity against FR-positive cells and tumors. EC72 was produced by coupling folic acid-gamma-cysteine to 7-N-modified MMC via a disulfide bond. This water soluble conjugate was found to retain high affinity for FR-positive cells, and it produced dose responsive activity in vitro against a panel of folate receptor (FR)-positive cell lines. EC72's activity was considered to be targeted and specific for the FR since (i) excess folic acid blocked biological activity, and (ii) FR-negative cell lines were unresponsive to this drug. Initial in vivo tests confirmed EC72's activity in both syngeneic and xenograft models, and this activity occurred in the apparent absence of gross or pathological toxicity. These results are significant, since daily dosing of EC72 for more than 30 consecutive days yielded no evidence of myelosuppression or toxicity to major organs, including the FR-positive kidneys. The latter observation supports published data, indicating that the apically oriented kidney proximal tubule FRs function to salvage folates prior to their excretion and to return these molecules back into systemic circulation. Overall, EC72's performance in vitro and in vivo warrants further preclinical study before this novel targeted chemotherapeutic is considered for clinical investigation.

The present study was to investigate whether metallothionein (MT) was involved in sensitivity of testis to cadmium (Cd) and protection of rats from Cd-induced testis damages. The rats were treated by intraperitoneal injection with 0.2, 0.4, and 0.8mg Cd/kg BW for 7 days. The atomic absorption spectrophotometry and cadmium hemoglobin affinity assay were applied to evaluate the contents of Cd and MT in testis and liver. The testis glutathione (GSH), malondialdehyde (MDA) and daily sperm production were measured. There were substantial increases of both Cd and MT in the liver after Cd exposure. The testis Cd and MT contents were lower than those in the corresponding liver in Cd-exposed rats. Low doses of Cd (0.2 and 0.4mg/kg BW) induced MT in testis, while a significant decline of MT was found in rats treated with 0.8mg Cd/kg BW. By a concomitant decrease of MT, there was an obvious increase of MDA and marked decreases of GSH, daily sperm production in rats treated with 0.8mg Cd/kg BW. These findings suggested MT was more difficult to be induced in the testis than in the liver by Cd, which might account for the high susceptibility of testis to Cd. MT, increased by a low dose of Cd, played an important role in protecting testis against Cd toxicity by sequestering and antioxidating.

Bacterial surfaces contain proteins, polysaccharides, and other biopolymers that can affect their adhesion to another surface. To better understand the role of proteins in bacterial adhesion, the interactions between two different model colloids (glass beads and carboxylated latex microspheres) and four proteins covalently bonded to glass surfaces were examined using colloid probes and an atomic force microscope (AFM). Adhesion forces between an uncoated glass colloid probe and protein-coated surfaces, measured in retraction force curves, decreased in the order poly-D-lysine > lysozyme > protein A > BSA. This ordering was consistent with the relative calculated charges of the proteins at neutral pH and the zeta-potentials measured for glass beads and latex microspheres coated with these proteins. When the glass bead was coated with a protein (BSA), overall adhesion forces between the protein-coated colloid and the protein-coated surfaces were reduced, and the adhesion force for each protein decreased in the same order observed in experiments with the uncoated glass bead. When latex colloid probes were coated with BSA, adhesion forces were significantly larger than those measured with BSA-coated glass colloid probes under the same conditions, demonstrating that the nature of the underlying colloid can affect the measured interaction forces. In addition, the adhesion forces measured with the BSA-coated latex colloid increased in a different order (BSA < lysozyme < protein A < poly-D-lysine) than that observed using the BSA-coated glass colloid. It was also found that increasing the solution ionic strength consistently decreased adhesion forces. This result is contrary to the general observation that bacterial adhesion increases with ionic strength. It was speculated that conformational changes of the protein produced this decrease in adhesion with increased ionic strength. These results suggest the need to measure nanoscale adhesion forces in order to understand better molecular scale interactions between colloids and surfaces.

Objective: This study was designed to examine the in vitro effects of fenvalerate on steroid production and steroidogenic enzymes mRNA expression level in rat granulosa cells., Methods: Using primary cultured rat granulosa cells (rGCs) as model, fenvalerate of various concentrations (0, 1, 5, 25, 125, 625 micromol/L) was added to the medium for 24 h. In some cases, optimal concentrations of 22(R)-hydroxycholesterol (25 micromol/L), Follicle stimulating hormone (FSH, 2 mg/L), or 8-Bromo-cAMP (1 mmol/L) were provided. Concentrations of 17 beta-estradiol(E2) and progesterone (P4) in the medium from the same culture wells were measured by RIA and the steroidogenic enzyme mRNA level was quantified by semi-quantitative RT-PCR., Results: Fenvalerate decreased both P4 and E2 production in a dose-dependent manner while it could significantly stimulate rGCs proliferation. This inhibition was stronger in the presence of FSH. Furthermore, it could not be reversed by 22(R)-hydroxycholesterol or 8-Bromo-cAMP. RT-PCR revealed that fenvalerate had no significant effect on 3 beta-HSD, but could increase the P450scc mRNA level. In addition, 17 beta-HSD mRNA level was dramatically reduced with the increase of fenvalerate dose after 24 h treatment., Conclusion: Fenvalerate inhibits both P4 and E2 production in rGCs. These results support the view that fenvalerate is considered as a kind of endocrine-disrupting chemicals. The mechanism of its disruption may involve the effects on steroidogenesis signaling cascades and/or steroidogenic enzyme's activity.

Aim: To evaluate the joint action of phoxim and fenvalerate on the reproductive function in male Sprague-Dawley rats., Methods: The 2 x 2 factorial analysis experiment was used in the study. The pesticides were orally given at daily doses of phoxim (Pho) 8.2 mg/kg, fenvalerate (Fen) 3.3 mg/kg and Pho 8.2+Fen 3.3 mg/kg (Pho:Fen = 5:2), 5 days a week for 60 days. Sperm motility was measured with computer-assisted sperm motility analysis (CASA) and daily sperm production estimated. Immunoenzymatic method and electron microscopy were used to evaluate the serum testosterone and the testicular morphology, respectively., Results: There were significant decreases in sperm motility parameters in the treated animals, including straight line velocity (VSL), beat cross frequency (BCF), linearity (LIN) and straightness (STR). After treated with Fen, significant decreases in VSL, LIN and STR were demonstrated. Significant decreases of daily sperm production were seen in animals treated with Pho and Pho+Fen in comparison with the controls. Serum testosterone levels were not significantly changed in the treated groups. Factorial ANOVA showed that no significant interactions were noted between Pho and Fen in sperm motility, sperm production and serum testosterone. Both the single and mixed pesticides caused various degrees of testicular lesions, involving vacuolation of endoplasmic reticulum and necrosis of Sertoli cells., Conclusion: The pesticides may cause sperm motility changes and testicular lesions in male rats. The action of Pho and Fen may be additive.

Aims: To determine sperm nuclear DNA integrity and to investigate the relation between fenvalerate (FE) exposure and spermatozoa DNA damage., Methods: Sperm DNA fragmentation was detected by a modified alkaline single cell gel electrophoresis (Comet) assay and a terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) assay. The olive tail moment (OTM) and percentage tail DNA were measured by the Comet assay, and cell positive percentage was measured by the TUNEL assay for DNA damage evaluation., Results: The DNA integrity of spermatozoa of external and internal control groups were both significantly greater than that of the FE exposed group. The median value of tail DNA percentage in the exposure group was 11.30, which was significantly higher than 5.60 in the internal control group and 5.10 in the external control group. The median value of OTM was 3.80 in the exposure group, significantly higher than 1.50 in the internal control group and 2.00 in the external control group. Mean cell positive was 31.2% in the exposure group, significantly higher than 17.4% in the internal control and 19.6% in the external control groups. Cell positive (%) was significantly correlated with tail DNA percentage and with OTM of whole subjects (n = 63)., Conclusions: Results showed that occupational FE exposure is associated with an increase in sperm DNA damage. A combination of the Comet and TUNEL assays would offer more comprehensive information for a better understanding of sperm DNA damage, and the biological significance of sperm DNA damage in sperm function and male infertility.

Unlabelled: Our group previously reported on the synthesis and characterization of a novel (99m)Tc-based folate-peptide chelator called EC20. This agent was found to bind folate receptor (FR)-positive cells and tissues with high affinity and was deemed useful for radiodiagnostic applications. In this study, we investigated the effect of D-amino acid substitution within EC20 on its tissue biodistribution. Expanded in vivo studies were also performed with unmodified (99m)Tc-EC20 to determine the effect of tumor FR expression, tumor size, tumor location, route of dose administration, and rodent diet on the agent's tissue biodistribution pattern., Methods: EC20 and EC53, the all-D-isomer of EC20, were synthesized and radiolabeled with (99m)Tc. The relative affinity of EC53 to the FR with respect to EC20 was then determined in cultured tumor cells. The ability of (99m)Tc-EC20 and (99m)Tc-EC53 to target tumors in vivo was examined using BALB/c mice with subcutaneously inoculated M109 or 4T1 cells, yielding 0.1- to 0.5-g tumors in 20 d., Results: The D-amino acid substitutions of EC20 were found to reduce the uptake of the agent into tumor and major organs. Subsequent studies using the original (99m)Tc-EC20 agent confirmed that its net tumor uptake was specific and proportional to FR expression levels in tumor cells as well as linear with respect to the overall tumor size. Further, (99m)Tc-EC20 uptake was found to be independent of both solid tumor location (intraperitoneal vs. subcutaneous) and the route of administration (intraperitoneal vs. intravenous). Interestingly, leucovorin supplementation of a commonly used folate-deficient laboratory chow had no effect on the agent's overall tissue biodistribution pattern. But, tumor-to-nontumor ratios could be increased up to 2.7-fold when 1 equivalent of free folic acid was coinjected with (99m)Tc-EC20., Conclusion: Taken together, these results confirm that (99m)Tc-EC20 has the potential to be a clinically useful noninvasive radiodiagnostic agent for detecting the locus of FR-positive cancers.