Pharmacogenomics of the human ABC transporter ABCG2: from functional evaluation to drug molecular design

In the post-genome-sequencing era, emerging genomic technologies are shifting the paradigm for drug discovery and development. Nevertheless, drug discovery and development still remain high-risk and high-stakes ventures with long and costly timelines. Indeed, the attrition of drug candidates in preclinical and development stages is a major problem in drug design. For at least 30% of the candidates, this attrition is due to poor pharmacokinetics and toxicity. Thus, pharmaceutical companies have begun to seriously re-evaluate their current strategies of drug discovery and development. In that light, we propose that a transport mechanism-based design might help to create new, pharmacokinetically advantageous drugs, and as such should be considered an important component of drug design strategy. Performing enzyme- and/or cell-based drug transporter, interaction tests may greatly facilitate drug development and allow the prediction of drug–drug interactions. We recently developed methods for high-speed functional screening and quantitative structure–activity relationship analysis to study the substrate specificity of ABC transporters and to evaluate the effect of genetic polymorphisms on their function. These methods would provide a practical tool to screen synthetic and natural compounds, and these data can be applied to the molecular design of new drugs. In this review article, we present an overview on the genetic polymorphisms of human ABC transporter ABCG2 and new camptothecin analogues that can circumvent AGCG2-associated multidrug resistance of cancer.     

Impact of FEBRA (fermented brown rice with Aspergillus oryzae) intake and concentrations of PCDDs, PCDFs and PCBs in blood of humans from Japan

Impact of FEBRA (fermented brown rice with Aspergillus oryzae, a rich dietary fiber) intake on the concentrations of polychlorinated-biphenyls (PCBs), -dioxin-like polychlorinated biphenyls (DLPCBs), -dibenzo-p-dioxins (PCDDs), and -dibenzofurans (PCDFs) in blood of nine married Japanese were studied for 2 years. Concentrations of total PCBs (average+/-standard deviation) were greater 326,000+/-263,000 and 207,000+/-89,000 in FEBRA-intake and non-FEBRA-intake groups, respectively, on pg/g lipid (from now onwards data expressed on pg/g lipid). DLPCBs were second major contaminants 94,000+/-77,000 and 60,000+/-28,000, orderly in FEBRA-intake and non-FEBRA-intake groups. The concentrations of PCDD/DFs were several orders of magnitude lower than those of PCBs, whereas PCDDs were greater (433+/-233 and 512+/-511, in FEBRA-intake and non-FEBRA-intake groups, respectively) than PCDFs (56+/-50 and 43+/-35). Calculated toxic equivalency (TEQ) concentrations were 34+/-24 and 23+/-10 in FEBRA-intake and non-FEBRA-intake groups, respectively. In order to corroborate the impact of FEBRA, the changes of PCDD/DFs and PCB concentrations for 0.5-year, 1-year, 1.5-year, and 2-year samples were estimated from average concentration of 1-day before study commences and 1-week study samples. The results explicit FEBRA-intake group eliminated 7%, 33%, 45%, 36%, and 26% and non-FEBRA-intake group eliminated 8%, 28%, 36%, 31%, and 20% of DLPCBs, PCDDs, PCDFs, PCDD/DF TEQ and total TEQ, respectively. This study is the first of its kind showing the possible elimination of PCDDs, PCDFs and their TEQ in humans from Japan after the consumption of FEBRA.     

Waste wood recycling as animal bedding and development of bio-monitoring tool using the CALUX assay

Animal bedding made of waste wood samples from seven different plants in Japan were chemically analyzed in terms of persistent organic pollutants (POPs) including polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/DFs), coplanar polychlorinated biphenyls (Co-PCBs), drin compounds, chlordane compounds and various inorganic toxic compounds (Cr, Cu, As, B, Cd and Pb) to investigate the chemical characteristics and levels of contamination. Further investigation was conducted to determine the success of applying the Chemically Activated Luciferase Expression (CALUX) bioassay to the waste wood samples in combination with a cleanup procedure for the detection of dioxin-like compounds in order to develop the CALUX bioassay as a rapid and cost-effective screening/monitoring method and a contributive tool to risk management in the waste wood recycling process. For the cleanup procedure, crude extracts from wood samples were prepared by dimethylsulfoxide (DMSO)/n-hexane extraction, and then the extracts were processed by silica gel-44% sulfuric acid reflux treatment at 70 degrees C for 60 min to yield the bioassay fractions. The presence of POPs and inorganic toxic compounds were confirmed in most of the litter samples. In particular, Co-PCBs in one sample (litter dust) showed a high concentration level (1200000 pg/g, 240 pg TEQ/g), suggesting the potential for contamination from demolition waste. The CALUX assay-determined TEQs (CALUX-TEQs) were significantly high in the sample after DMSO/n-hexane extraction, probably due to labile aryl hydrocarbon receptor (AhR) ligands such as PAHs; however, they were remarkably reduced through a single silica gel-44% sulfuric acid reflux treatment. The ratio between CALUX-TEQ values and WHO toxicity equivalent values (WHO-TEQ) obtained by congener-specific chemical analysis ranged from 0.058 to 22 and show comparatively good agreement. Underestimation in some samples, however, was observed where WHO-TEQ values of Co-PCBs contributed greatly to total WHO-TEQ values. Reasons for this gap could be lower CALUX assay-determined relative potencies (REPs) than the WHO-TEFs for these congeners or AhR-antagonistic effects of non dioxin-like PCBs which coexist at higher concentration than Co-PCBs. The CALUX assay is proposed as a promising application in the recycling process of wooden materials.     

Detection and average content levels of carcinogenic and mutagenic compounds from the particulates on diesel and gasoline engine mufflers

Carcinogenic and mutagenic compounds, which were extracted from the particulates that adhered to inner surfaces of diesel and gasoline engine mufflers, were quantified by the series method of Soxhlet extraction, liquid-liquid partition, thin-layer chromatography, and spectrofluorometry. Mutagenic activity of their neutral and acidic fractions was tested in the improved Ames assay by the preincubation method with Salmonella typhimurium TA98 in the presence and absence of metabolic activation system (S-9 mix). The average content levels (μg/g tar) of polycyclic aromatic hydrocarbons from gasoline engine cars were greater than those from diesel engine vehicles. However, the levels of nitro derivatives of PAHs and polycyclic quinones from the diesel engines were greater than from the gasoline engines. Mutagenic activity of the diesel acidic fraction was the highest among the diesel and gasoline fractions, and was significantly higher in the absence of the S-9 mix. Furthermore, the relative value (R_{c = 0}) of infrared absorption of carbonyl stretching vibration to that of methylene asymmetric stretching vibration of the diesel acidic fraction was the highest among the diesel and gasoline fractions. These results strongly suggest that highly direct-acting mutagens in the acidic fraction are at higher levels in diesel emission particulates than those from gasoline, and that these mutagens are carboxylic acid, aldehyde, and alcohol derivatives of PAHs and NPAHs.     

Detonation characteristics of ammonium nitrate and activated carbon mixtures

To better understand the detonation characteristics of ammonium nitrate (AN) and activated carbon (AC) mixtures, steel tube tests were carried out for AN/AC mixtures of various compositions and different forms of AN (powdered, prilled, phase stabilized and granular), and the detonation velocity was measured. The powdered AN/AC mixtures gave higher detonation velocities than the other AN forms. For all the AN/AC mixtures, the experimentally observed detonation velocities at each loading density were far below the theoretically predicted values calculated by the CHEETAH code based on thermohydrodynamics, exhibiting so-called non-ideal detonation. The lowest detonation velocity of powdered AN/AC mixtures was obtained as D=1.25 km/s for an AC content of 0.1 wt%. This was considered to be close to the critical condition for stable detonation.     

Substance flow analysis of coplanar PCBs released from waste incineration processes

Previous reports have focused on the emission of coplanar polychlorinated biphenyls (Co-PCBs) which have a toxic mechanism similar to that of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDDs/DFs) released from municipal solid waste (MSW) incineration. Such emissions accounted for a small percentage of all the dioxins (PCDDs/DFs and Co-PCBs) recorded at the toxicity equivalent (TEQ) level. There is, however, very little information about Co-PCBs, such as the quantities being released and their effect on overall environmental pollution. The aim of this research has been to clarify the substance flow of Co-PCBs from MSW incineration processes. The results reveal that whereas the input of Co-PCBs into the MSW incineration facilities in Kyoto City was 0.13–0.29 μg-TEQ per ton waste, the total output of Co-PCBs (the sum of Co-PCBs released from emission gas, fly ash, and bottom ash) was 4.9 μg-TEQ per ton waste. The total output was therefore found to be higher than the total input. Over 90% of the total PCBs were decomposed in the incineration process. In comparing the profiles of congeners and homologues, those in the MSW were found to be similar to those detected in the atmosphere and products containing PCBs, but different from those in the MSW incineration gas. Received: August 26, 1998 / Accepted: March 2, 1999     

Contamination,dynamics, and health risk assessment of pesticides in seawater and marine samples from the Seto Inland Sea,Japan

Environmental Science and Pollution Research - We assessed the contamination, dynamics, and health risks of the pesticides cyanazine, simetryn, fenarimol, isoprothiolane, diazinon, irgarol,...     

Remediation of toxic metal contaminated soil by washing with biodegradable aminopolycarboxylate chelants

Ex situ soil washing with synthetic extractants such as, aminopolycarboxylate chelants (APCs) is a viable treatment alternative for metal-contaminated site remediation. EDTA and its homologs are widely used among the APCs in the ex situ soil washing processes. These APCs are merely biodegradable and highly persistent in the aquatic environments leading to the post-use toxic effects. Therefore, an increasing interest is focused on the development and use of the eco-friendly APCs having better biodegradability and less environmental toxicity. The paper deals with the results from the lab-scale washing treatments of a real sample of metal-contaminated soil for the removal of the ecotoxic metal ions (Cd, Cu, Ni, Pb, and Zn) using five biodegradable APCs, namely [S,S]-ethylenediaminedisuccinic acid, imminodisuccinic acid, methylglycinediacetic acid, DL-2-(2-carboxymethyl) nitrilotriacetic acid (GLDA), and 3-hydroxy-2,2′-iminodisuccinic acid. The performance of those biodegradable APCs was evaluated for their interaction with the soil mineral constituents in terms of the solution pH and metal-chelant stability constants, and compared with that of EDTA. Speciation calculations were performed to identify the optimal conditions for the washing process in terms of the metal-chelant interactions as well as to understand the selectivity in the separation ability of the biodegradable chelants towards the metal ions. A linear relationship between the metal extraction capacity of the individual chelants towards each of the metal ions from the soil matrix and metal-chelant conditional stability constants for a solution pH greater than 6 was observed. Additional considerations were derived from the behavior of the major potentially interfering cations (Al, Ca, Fe, Mg, and Mn), and it was hypothesized that use of an excess of chelant may minimize the possible competition effects during the single-step washing treatments. Sequential extraction procedure was used to determine the metal distribution in the soil before and after the extractive decontamination using biodegradable APCs, and the capability of the APCs in removing the metal ions even from the theoretically immobilized fraction of the contaminated soil was observed. GLDA appeared to possess the greatest potential to decontaminate the soil through ex situ washing treatment compared to the other biodegradable chelants used in the study.     

Possible change in Asian dust source suggested by atmospheric anthropogenic radionuclides during the 2000s

Decades-long monitoring of anthropogenic radionuclides in the atmospheric deposition in Tsukuba, Japan suggests not only the substantial impacts of the Asian dust (Kosa) on the deposition but also the possible change of the Kosa source region, especially during springs of the 2000s. In order to know more about such change, 4 single wet deposition events occurred in the spring of 2007 were scrutinized. The largest anthropogenic radionuclides wet deposition was supplied by the April 2–4 event. It brought several tens % of the monthly depositions (April 2007) of the dust (residue) mass (4.5 g m^?2) and anthropogenic radionulides (⁹⁰Sr: 16, ¹³⁷Cs: 97 and Pu: 3 mBq m^?2). None of the events observed fulfilled both criteria of the specific activities and ⁹⁰Sr/¹³⁷Cs activity ratio to the Tsukuba soil; they did not exhibit local soil dust signature. The Kosa events in fact have extensive impacts on the atmospheric environment over Japan in spring season. Considering the elevated specific activities as well as greater ¹³⁷Cs/⁹⁰Sr activity ratio in the deposited dust, it is hypothesized that the dust source areas in Asian continent would be shifting from the arid zone to the desert-steppe zone suffering from desertification during the 2000s. This type of the Kosa may be called as the ‘new-regime Kosa’. Chemical observation in the far downwind region of the Kosa dust could allow us to know possible shift in the source regions.     

Evaluating the strength development of mortar using clinker fine aggregate with a combination of fly ash and its inhibitory effects on alkali–silica reaction and delayed ettringite formation

The aggregate composed of cement clinker, an intermediate cement product, improves strength development and mass transfer resistance of concrete. Fly ash (FA) is a supplementary cementitious material that can be substituted by cement. This study investigated the strength development of highly FA-substituted mortar mixed with cement clinker fine aggregate (CL) and tested its inhibitory effect against the alkali–silica reaction (ASR). In addition to these, this study provides the testing results of evaluating the effects of using cement clinker fine aggregate on delayed ettringite formation (DEF), which can be problematic for precast concrete products. The study results revealed that at 91 days of age, in the case of 80% replacement ratio of fly ash to cement, using CL exhibited similar strength development as mortar with limestone fine aggregate and no FA substitution. Furthermore, mortars with 70% and 80% FA substitution did not exhibit clear ASR-induced expansion even at 182 days of age. Lastly, mortar using clinker fine aggregate could suppress DEF-induced expansion at 182 days of age, which was similar to the effect obtained using FA. These results can promote the utilization of CL resources for concrete and using clinker as fine aggregate in precast concrete products.