Using live videography observation and Bayesian isotope mixing model to identify food composition and dietary contribution to inorganic mercury and methylmercury intake by songbird nestlings.

Mercury (Hg) exposure is increasing in terrestrial birds; however, studies on its sources are scarce. In the present study, we elucidated the food composition of green-backed tit nestlings from three urban forest parks (CPL, AHL, and LCG) using live videography observation (LVO). Furthermore, the daily dietary intakes of inorganic Hg (IHg) (MDIIHg) and methylmercury (MeHg) (MDIMeHg) were determined using the Bayesian isotope mixing model (BIMM) to uncover the nestlings' specific dietary Hg contribution. Both LVO and BIMM indicated that Lepidoptera (primarily caterpillar) constituted the primary food source for the nestlings in the three forests, accounting for approximately 60% of their diet in all three forest parks. The estimated MDI of Hg revealed that lepidopterans and spiders primarily contributed to IHg exposure, with a co-contribution ratio of 71.8%-97.7%. Unexpectedly, dietary MeHg was mostly derived from spiders; the highest contribution ratio of 93.6% was recorded at CPL, followed by another peak ratio of 92.9% at LCG. However, the dietary exposure was primarily IHg, accounting for 69.8% (AHL), 62.0% (LCG), and 61.3% (CPL) of the nestlings. Our study findings highlight the importance of dietary IHg transfer in evaluating the effects of Hg in nestlings. LVO, coupled with BIMM, is an effective tool for determining the food compositions of songbird nestlings and estimating the contribution of specific diets.

Home-produced eggs: An important pathway of methylmercury exposure for residents in mercury mining areas, southwest China.

In light of the documented elevated concentrations of total mercury (Hg) and methylmercury (MeHg) in poultry originating from Hg-contaminated sites, a knowledge gap persists regarding the levels of Hg found in home-produced eggs (HPEs) and the associated dietary exposure risks in regions affected by Hg mining. To address this knowledge gap, a comprehensive investigation was undertaken with the primary objectives of ascertaining the concentrations of THg and MeHg in HPEs and evaluating the potential hazards associated with the consumption of eggs from the Wanshan Hg mining area in Southwest China. The results showed that THg concentrations in HPEs varied within a range of 10.5-809 ng/g (with a geometric mean (GM) of 64.1 ± 2.7 ng/g), whereas MeHg levels spanned from 1.3 to 291 ng/g (GM, 23.1 ± 3.4 ng/g). Remarkably, in half of all eggs, as well as those collected from regions significantly impacted by mining activities, THg concentrations exceeded the permissible maximum allowable value for fresh eggs (50 ng/g). Consumption of these eggs resulted in increased exposure risks associated with THg and MeHg, with GM values ranging from 0.024 to 0.17 µg/kg BW/day and 0.0089-0.066 µg/kg BW/day, respectively. Notably, the most substantial daily dosage was observed among children aged 2-3 years. The study found that consuming HPEs could result in a significant IQ reduction of 34.0 points for the whole mining area in a year. These findings highlight the potential exposure risk, particularly concerning MeHg, stemming from the consumption of local HPEs by residents in mining areas, thereby warranting serious consideration within the framework of Hg exposure risk assessment in mining locales.

Enhanced Mercury Accumulation in Riparian Spiders: An Evidence of Insects' Emergence Effect in Aquatic and Upland Terrestrial Crossed Habitat.

Many studies have focused on mercury (Hg) accumulation in both aquatic and terrestrial organisms, but the effects of aquatic Hg on terrestrial organisms have rarely been documented. Here we report the accumulation of Hg in two species of spiders, Argiope bruennichi, inhabiting paddy fields, and Nephila clavata, inhabiting small forests in the riparian zones of two hydroelectric reservoirs in Guiyang, southwest China. The mean concentration of total mercury (THg) was higher in N. clavata (0.38 mg kg-1) than in A. bruennichi (0.20 mg kg-1). The monthly average THg in N. clavata, collected consecutively from May to October, and the highest values for THg in June (1.2 mg kg-1) could be related to the emergence of aquatic insects during early summer, suggesting that emerging insects play a crucial role in the accumulation of Hg in riparian spiders. The high values could also be attributable to the different times of spider sampling or individual differences.

Total and methylmercury concentrations in nocturnal migratory birds passing through Mount Ailao, Southwest China.

Despite growing concerns over mercury (Hg) accumulation in birds in recent decades, little is known about Hg exposure in nocturnal migratory birds. Here, total mercury (THg) and methylmercury (MeHg) were detected in the feathers of nocturnal migratory birds (n = 286, belonging to 46 species) passing through Mount Ailao in Southwest China. The stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) were also determined to clarify the effects of trophic position, foraging guild, and foraging behavior on Hg bioaccumulation. Our results show that the THg and MeHg concentrations varied by two orders of magnitude among all nocturnal migratory birds investigated, with the lowest values (THg: 0.056 mg kg-1; MeHg: 0.038 mg kg-1) in the Asian koel (Eudynamys scolopaceus) and the highest (THg: 12 mg kg-1; MeHg: 7.8 mg kg-1) in the hair-crested drongo (Dicrurus hottentottus). Waterbirds showed higher δ15N values and higher THg and MeHg concentrations than songbirds, and the Hg concentrations in piscivorous species were significantly higher than those in herbivores, omnivores, and insectivores. Significant effects of foraging guilds (Kruskal-Wallis one-way ANOVA, p < 0.001) and foraging behaviors (Kruskal-Wallis one-way ANOVA, p < 0.001) on the Hg concentrations in migratory bird feathers were detected. A risk assessment indicated that approximately 7.0% of individuals were at moderate (2.4-5.0 mg kg-1) to high (>5.0 mg kg-1) risk of Hg exposure, and were therefore vulnerable to adverse physiological and behavioral effects. A long-term monitoring campaign during the migratory period is highly recommended to better understand the bioaccumulation of Hg in these nocturnal migratory bird populations over time.

Extremely Elevated Total Mercury and Methylmercury in Forage Plants in a Large-Scale Abandoned Hg Mining Site: A Potential Risk of Exposure to Grazing Animals.

Ninety-five wild forage plants (belonging to 22 species of 18 families) and their corresponding rhizosphere soil samples were collected from wastelands of a large-scale abandoned Hg mining region for total Hg (THg) and methylmercury (MeHg) analysis. The forage plant communities on the wastelands were dominated by the Asteraceae, Crassulaceae, and Polygonaceae families. The THg and MeHg concentrations in the forage plants varied widely and were in the range of 0.10 to 13 mg/kg and 0.19 to 23 µg/kg, respectively. Shoots of Aster ageratoides showed the highest average THg concentration of 12 ± 1.1 mg/kg, while those of Aster subulatus had the highest average MeHg concentrations of 7.4 ± 6.1 µg/kg. Both the THg and MeHg concentrations in the aboveground plant parts exhibited positive correlations with the THg (r = 0.70, P < 0.01) and MeHg (r = 0.68, P < 0.01) concentrations in the roots; however, these were not correlated with the THg and MeHg concentrations in their rhizosphere soils. The species A. ageratoides, A. subulatus, and S. brachyotus showed strong accumulation of Hg and are of concern for herbivorous/omnivorous wildlife and feeding livestock. Taking the provisional tolerable weekly intake (PTWI) values for IHg recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA in Summary and conclusions of the seventy-second meeting of the joint FAO/WHO expert committee on food additives Rome, Italy, 2010) for human dietary exposure of 4 ng/g into account, the daily intake of IHg by a 65 kg animal grazing on 1.0 kg of forage (dry weight) would be between 190 and 13,200 µg, three to five orders of magnitude higher than the permitted limit, suggesting a potential risk of exposure.

Distributions of Total Mercury and Methylmercury in Dragonflies from a Large, Abandoned Mercury Mining Region in China.

Dragonflies (Order Odonata) often are considered to be biosentinels of environmental contamination, e.g., heavy metals and/or persistent organic pollutants (POPs). Dragonflies (n = 439) belonging to 15 species of 8 genera were collected from an abandoned mercury (Hg) mining region in China to investigate the bioaccumulation of total Hg (THg) and methylmercury (MeHg). THg and MeHg concentrations in dragonflies varied widely within ranges of 0.06-19 mg/kg (average: 1.5 ± 2.2 mg/kg) and 0.02-5.7 mg/kg (average: 0.75 ± 0.65 mg/kg), respectively. THg and MeHg were positively correlated with bodyweight (THg: r2 = 0.10, P = 0.000; MeHg: r2 = 0.09, P = 0.000). Significant variations were observed among species, with the highest MeHg value (in Orthetrum triangulare) was fivefold higher than the lowest (in Pantala flavescens). These variations were consistent with those of nitrogen isotope (δ15N) values, indicating that increased δ15N, i.e., trophic levels, may reflect increased exposure and uptake of biomagnifying MeHg in dragonflies. A toxicological risk assessment found hazard quotients for specialist dragonfly-consuming birds of up to 7.2, which is 2.4 times greater than the permissible limit of 3, suggesting a potential toxicological risk of exposure.

Inhibition of cyclin-dependent kinase 7 down-regulates yes-associated protein expression in mesothelioma cells.

Cyclin-dependent kinase 7 (CDK7) is a protein kinase that plays a major role in transcription initiation. Yes-associated protein (YAP) is a main effector of the Hippo/YAP signalling pathway. Here, we investigated the role of CDK7 on YAP regulation in human malignant pleural mesothelioma (MPM). We found that in microarray samples of human MPM tissue, immunohistochemistry staining showed correlation between the expression level of CDK7 and YAP (n = 70, r = .513). In MPM cells, CDK7 expression level was significantly correlated with GTIIC reporter activity (r = .886, P = .019). Inhibition of CDK7 by siRNA decreased the YAP protein level and the GTIIC reporter activity in the MPM cell lines 211H, H290 and H2052. Degradation of the YAP protein was accelerated after CDK7 knockdown in 211H, H290 and H2052 cells. Inhibition of CDK7 reduced tumour cell migration and invasion, as well as tumorsphere formation ability. Restoration of the CDK7 gene rescued the YAP protein level and GTIIC reporter activity after siRNA knockdown in 211H and H2052 cells. Finally, we performed a co-immunoprecipitation analysis using an anti-YAP antibody and captured the CDK7 protein in 211H cells. Our results suggest that CDK7 inhibition reduces the YAP protein level by promoting its degradation and suppresses the migration and invasion of MPM cells. Cyclin-dependent kinase 7 may be a promising therapeutic target for MPM.

Synthesis of Sulfonylated Lactams by Copper-Mediated Aminosulfonylation of 2-Vinylbenzamides with Sodium Sulfinates.

Treatment of 2-vinylbenzamide derivatives with sulfinate sodium in the presence of Cu(NO3)2·3H2O led to an intra/intermolecular aminosulfonylation reaction to produce sulfonylated lactams in moderate to good yields. The developed method features the easily available and stable sulfone reagents, ease of operation, and a broad functional group tolerance.

Radiosynthesis of a carbon-11-labeled AMPAR allosteric modulator as a new PET radioligand candidate for imaging of Alzheimer's disease.

To develop PET tracers for imaging of Alzheimer's disease, a new carbon-11-labeled AMPAR allosteric modulator 4-cyclopropyl-7-(3-[11C]methoxyphenoxy)-3,4-dihydro-2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide ([11C]8) has been synthesized. The reference standard 4-cyclopropyl-7-(3-methoxyphenoxy)-3,4-dihydro-2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide (8) and its corresponding desmethylated precursor 4-cyclopropyl-7-(3-hydroxyphenoxy)-3,4-dihydro-2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide (9) were synthesized from 4-methoxyabiline and chlorosulfonyl isocyanate in eight and nine steps with 3% and 1% overall chemical yield, respectively. The target tracer [11C]8 was prepared from the precursor 9 with [11C]CH3OTf through O-[11C]methylation and isolated by HPLC combined with SPE in 10-15% radiochemical yield, based on [11C]CO2 and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the molar activity (AM) at EOB was 370-740 GBq/µmol with a total synthesis time of 35-40-minutes from EOB.

Facile synthesis of carbon-11-labeled sEH/PDE4 dual inhibitors as new potential PET agents for imaging of sEH/PDE4 enzymes in neuroinflammation.

To develop PET tracers for imaging of neuroinflammation, new carbon-11-labeled sEH/PDE4 dual inhibitors have been synthesized. The reference standard N-(4-methoxy-2-(trifluoromethyl)benzyl)benzamide (1) and its corresponding desmethylated precursor N-(4-hydroxy-2-(trifluoromethyl)benzyl)benzamide (2) were synthesized from (4-methoxy-2-(trifluoromethyl)phenyl)methanamine and benzoic acid in one and two steps with 84% and 49% overall chemical yield, respectively. The standard N-(4-methoxy-2-(trifluoromethyl)benzyl)-1-propionylpiperidine-4-carboxamide (MPPA, 4) and its precursor N-(4-hydroxy-2-(trifluoromethyl)benzyl)-1-propionylpiperidine-4-carboxamide (5) were synthesized from methyl 4-piperidinecarboxylate, propionyl chloride and (4-methoxy-2-(trifluoromethyl)phenyl)methanamine in two and three steps with 62% and 34% overall chemical yield, respectively. The target tracers N-(4-[11C]methoxy-2-(trifluoromethyl)benzyl)benzamide ([11C]1) and N-(4-[11C]methoxy-2-(trifluoromethyl)benzyl)-1-propionylpiperidine-4-carboxamide ([11C]MPPA, [11C]4) were prepared from their corresponding precursors 2 and 5 with [11C]CH3OTf through O-[11C]methylation and isolated by HPLC combined with SPE in 25-35% radiochemical yield, based on [11C]CO2 and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the molar activity (AM) at EOB was 370-740 GBq/µmol with a total synthesis time of 35-40-minutes from EOB.

Cadmium contamination in a soil-rice system and the associated health risk: An addressing concern caused by barium mining.

Cadmium (Cd) is associated with barite; however, its biogeochemical characteristics in environments impacted by barium (Ba) mining are not known. Here, we first revealed the characteristics of Cd concentrations, distributions, and chemical forms in the soil-rice system in Ba mining areas. The associated exposure and risk assessments of Cd via rice consumption were also conducted. Elevated levels of Cd with a wide range of 0.054-91 mg/kg were found in paddy soils, approximately 63% of which exceeded the national Grade II value for soil Cd levels in China (0.3 mg/kg). A significant positive correlation between the soil Cd and soil Ba demonstrated that large amounts of Cd were released into the environment from Ba mining. Cadmium accumulated remarkably in the rice grains (0.007-3.5 mg/kg). The chemical forms in the rice plants indicated that most of the Cd was in the pectate/protein fraction (F2, 92% in the grains and 61-71% in the other tissues), followed by the residual fraction (F3, 7.1% in the grains, 27-38% in the other tissues). A minor portion of Cd was in the soluble and aminophenol fraction (F1, 0.44% in the grains, 0.26-1.4% in the other tissues). The positive correlations observed between the grain Cd and F2 in the roots, stems and leaves suggested that Cd in the rice grain was mainly from F2. Similarly, the root F2 was also positively correlated with that in the stems/leaves, indicating the critical role of F2 in Cd2+ migration in rice tissues. The estimated average hazard quotient (2.5) and annual excess lifetime cancer risk (21 × 10-5 a-1) were higher than the safety levels of 1 and 5.0 × 10-5 a-1, respectively, showing that the dietary intake of Cd via rice consumption posed high health risks to residents. Our study demonstrated that more concerns should be paid to Cd contamination in Ba mining areas.

Effects of Selenium on Mercury Bioaccumulation in a Terrestrial food Chain from an Abandoned Mercury Mining Region.

Few reports of the relationship exist between mercury (Hg) and selenium (Se) from locations of severe Hg contamination in terrestrial environments. Here, we report the concentrations of Hg and Se as well as Se:Hg molar ratios in biotic samples collected from a region with a long history of Hg mining. Nitrogen isotopes (δ15N) were analyzed to confirm the trophic levels. Results showed that bird feathers at the top trophic level exhibited the highest Hg concentrations, while the lowest concentrations were found in herbivorous insects, demonstrating a significant biomagnification across the food chain. In contrast, herbivorous insects of different types (generalists vs. specialized rice pests) exhibited both the highest and the lowest concentrations of Se, indicating a lack of biomagnification. Indeed, Se was correlated positively with Hg when Se:Hg ratios were greater than one, suggesting Se:Hg molar ratios can be a controlling influence on Hg in terrestrial organisms.

Inhibition of yes-associated protein suppresses brain metastasis of human lung adenocarcinoma in a murine model.

Yes-associated protein (YAP) is a main mediator of the Hippo pathway and promotes cancer development and progression in human lung cancer. We sought to determine whether inhibition of YAP suppresses metastasis of human lung adenocarcinoma in a murine model. We found that metastatic NSCLC cell lines H2030-BrM3(K-rasG12C mutation) and PC9-BrM3 (EGFRΔexon19 mutation) had a significantly decreased p-YAP(S127)/YAP ratio compared to parental H2030 (K-rasG12C mutation) and PC9 (EGFRΔexon19 mutation) cells (P < .05). H2030-BrM3 cells had significantly increased YAP mRNA and expression of Hippo downstream genes CTGF and CYR61 compared to parental H2030 cells (P < .05). Inhibition of YAP by short hairpin RNA (shRNA) and small interfering RNA (siRNA) significantly decreased mRNA expression in downstream genes CTGF and CYR61 in H2030-BrM3 cells (P < .05). In addition, inhibiting YAP by YAP shRNA significantly decreased migration and invasion abilities of H2030-BrM3 cells (P < .05). We are first to show that mice inoculated with YAP shRNA-transfected H2030-BrM3 cells had significantly decreased metastatic tumour burden and survived longer than control mice (P < .05). Collectively, our results suggest that YAP plays an important role in promoting lung adenocarcinoma brain metastasis and that direct inhibition of YAP by shRNA suppresses H2030-BrM3 cell brain metastasis in a murine model.

Inhibition of yes-associated protein down-regulates PD-L1 (CD274) expression in human malignant pleural mesothelioma.

Although tumour PD-L1 (CD274) expression had been used as a predictive biomarker in checkpoint immunotherapy targeting the PD1/PD-L1 axis in various cancers, the regulation of PD-L1 (CD274) expression is unclear. Yes-associated protein (YAP), an important oncogenic protein in Hippo signalling pathway, reportedly promotes cancer development. We investigated whether inhibition of YAP down-regulates PD-L1 (CD274) in human malignant pleural mesothelioma (MPM). Western blotting showed that 2 human MPM cell lines (H2052 and 211H) had increased PD-L1 protein expression compared to H290, MS-1 and H28 cells. In H2052 and 211H cells, PD-L1 mRNA expression was significantly increased compared to other MPM cell lines; YAP knockdown by small interfering RNA decreased PD-L1 protein and mRNA expression. Forced overexpression of the YAP gene increased PD-L1 protein expression in H2452 cells. Chromatin immunoprecipitation (ChIP) assay showed the precipitation of PD-L1 enhancer region encompassing 2 putative YAP-TEAD-binding sites in H2052 cells. We found that, in human MPM tissue microarray samples, YAP and PD-L1 concurrently expressed in immunohistochemistry stain (n = 70, P < .05, chi-square). We conclude that PD-L1 is correlated with YAP expression, and inhibition of YAP down-regulates PD-L1 expression in human MPM. Further study of how YAP regulates PD-L1 in MPM is warranted.

Synthesis of carbon-11-labeled 5-HT6R antagonists as new candidate PET radioligands for imaging of Alzheimer's disease.

Carbon-11-labeled serotonin (5-hydroxytryptamine) 6 receptor (5-HT6R) antagonists, 1-[(2-bromophenyl)sulfonyl]-5-[11C]methoxy-3-[(4-methyl-1-piperazinyl)methyl]-1H-indole (O-[11C]2a) and 1-[(2-bromophenyl)sulfonyl]-5-methoxy-3-[(4-[11C]methyl-1-piperazinyl)methyl]-1H-indole (N-[11C]2a), 5-[11C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1-(phenylsulfonyl)-1H-indole (O-[11C]2b) and 5-methoxy-3-((4-[11C]methylpiperazin-1-yl)methyl)-1-(phenylsulfonyl)-1H-indole (N-[11C]2b), 1-((4-isopropylphenyl)sulfonyl)-5-[11C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1H-indole (O-[11C]2c) and 1-((4-isopropylphenyl)sulfonyl)-5-methoxy-3-((4-[11C]methylpiperazin-1-yl)methyl)-1H-indole (N-[11C]2c), 1-((4-fluorophenyl)sulfonyl)-5-[11C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1H-indole (O-[11C]2d) and 1-((4-fluorophenyl)sulfonyl)-5-methoxy-3-((4-[11C]methylpiperazin-1-yl)methyl)-1H-indole (N-[11C]2d), were prepared from their O- or N-desmethylated precursors with [11C]CH3OTf through O- or N-[11C]methylation and isolated by HPLC combined with SPE in 40-50% radiochemical yield, based on [11C]CO2 and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the molar activity (MA) at EOB was 370-740 GBq/µmol with a total synthesis time of ∼40-min from EOB.

Targeting YAP in malignant pleural mesothelioma.

Malignant mesothelioma is an aggressive cancer that is resistant to current therapy. The poor prognosis of mesothelioma has been associated with elevated Yes-associated protein (YAP) activity. In this study, we evaluated the effect of targeting YAP in mesothelioma. First, we comprehensively studied YAP activity in five mesothelioma cell lines (211H, H2052, H290, MS-1 and H2452) and one normal mesothelial cell line (LP9). We found decreased phospho-YAP to YAP protein ratio and consistently increased GTIIC reporter activity in 211H, H2052 and H290 compared to LP9. The same three cell lines (IC50 s < 1 µM) were more sensitive than LP9 (IC50 = 3.5 µM) to the YAP/TEAD inhibitor verteporfin. We also found that verteporfin significantly reduced YAP protein level, mRNA levels of YAP downstream genes and GTIIC reporter activity in the same three cell lines, indicating inhibition of YAP signaling by verteporfin. Verteporfin also impaired invasion and tumoursphere formation ability of H2052 and H290. To validate the effect of specific targeting YAP in mesothelioma cells, we down-regulated YAP by siRNA. We found siYAP significantly decreased YAP transcriptional activity and impaired invasion and tumoursphere formation ability of H2052 and H290. Furthermore, forced overexpression of YAP rescued GTIIC reporter activity and cell viability after siYAP targeting 3'UTR of YAP. Finally, we found concurrent immunohistochemistry staining of ROCK2 and YAP (P < 0.05). Inhibition of ROCK2 decreased GTIIC reporter activity in H2052 and 211H suggesting that Rho/ROCK signaling also contributed to YAP activation in mesothelioma cells. Our results indicate that YAP may be a potential therapeutic target in mesothelioma.

Synthesis of [11C]HG-10-102-01 as a new potential PET agent for imaging of LRRK2 enzyme in Parkinson's disease.

The reference standard (4-((5-chloro-4-(methylamino)pyrimidin-2-yl)amino)-3-methoxyphenyl)(morpholino)methanone (HG-10-102-01) and its precursor (4-((5-chloro-4-(methylamino)pyrimidin-2-yl)amino)-3-hydroxyphenyl)(morpholino)methanone (desmethyl-HG-10-102-01) were synthesized from 2,4,5-trichloropyrimide and 3-methoxy-4-nitrobenzoic acid with overall chemical yield 49% in four steps and 14% in five steps, respectively. The target tracer (4-((5-chloro-4-(methylamino)pyrimidin-2-yl)amino)-3-[11C]methoxyphenyl)(morpholino)methanone ([11C]HG-10-102-01) was prepared from the precursor desmethyl-HG-10-102-01 with [11C]CH3OTf through O-[11C]methylation and isolated by HPLC combined with SPE in 45-55% radiochemical yield, based on [11C]CO2 and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the specific activity (SA) at EOB was 370-1110GBq/µmol with a total synthesis time of ∼40-min from EOB.

Knockdown of cullin 4A inhibits growth and increases chemosensitivity in lung cancer cells.

Cullin 4A (Cul4A) has been observed to be overexpressed in various cancers. In this study, the role of Cul4A in the growth and chemosensitivity in lung cancer cells were studied. We showed that Cul4A is overexpressed in lung cancer cells and tissues. Knockdown of the Cul4A expression by shRNA in lung cancer cells resulted in decreased cellular proliferation and growth in lung cancer cells. Increased sensitivity to gemcitabine, a chemotherapy drug, was also noted in those Cul4A knockdown lung cancer cells. Moreover, increased expression of p21, transforming growth factor (TGF)-ß inducible early gene-1 (TIEG1) and TGF beta-induced (TGFBI) was observed in lung cancer cells after Cul4A knockdown, which may be partially related to increased chemosensitivity to gemcitabine. G0/G1 cell cycle arrest was also noted after Cul4A knockdown. Notably, decreased tumour growth and increased chemosensitivity to gemcitabine were also noted after Cul4A knockdown in lung cancer xenograft nude mice models. In summary, our study showed that targeting Cul4A with RNAi or other techniques may provide a possible insight to the development of lung cancer therapy in the future.

Cul4A overexpression associated with Gli1 expression in malignant pleural mesothelioma.

Malignant pleural mesothelioma (mesothelioma) is a highly aggressive cancer without an effective treatment. Cul4A, a scaffold protein that recruits substrates for degradation, is amplified in several human cancers, including mesothelioma. We have recently shown that Cul4A plays an oncogenic role in vitro and in a mouse model. In this study, we analysed clinical mesothelioma tumours and found moderate to strong expression of Cul4A in 70.9% (51/72) of these tumours, as shown by immunohistochemistry. In 72.2% mesothelioma tumours with increased Cul4A copy number identified by fluorescence in situ hybridization analysis, Cul4A protein expression was moderate to strong. Similarly, Cul4A was overexpressed and Cul4A copy number was increased in human mesothelioma cell lines. Because Gli1 is highly expressed in human mesothelioma cells, we compared Cul4A and Gli1 expression in mesothelioma tumours and found their expression associated (P < 0.05, chi-square). In mesothelioma cell lines, inhibiting Cul4A by siRNA decreased Gli1 expression, suggesting that Gli1 expression is, at least in part, regulated by Cul4A in mesothelioma cells. Our results suggest a linkage between Cul4A and Gli1 expression in human mesothelioma.

Synthesis of a PET tau tracer [(11)C]PBB3 for imaging of Alzheimer's disease.

The authentic standard PBB3 and its precursor N-desmethyl-PBB3 as well as TBS-protected N-desmethyl-PBB3 precursor for radiolabeling were synthesized from 5-bromo-2-nitropyridine, acrolein diethyl acetal, 6-methoxy-2-methylbenzothiazole, and diethylchlorophosphate with overall chemical yield 1% in six steps, 2% in five steps, and 1% in six steps, respectively. [(11)C]PBB3 was prepared from either desmethyl-PBB3 or TBS-protected desmethyl-PBB3 with [(11)C]CH3OTf through N-[(11)C]methylation and isolated by HPLC combined with SPE in 20-25% and 15-20% radiochemical yield, respectively, based on [(11)C]CO2 and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the specific activity at EOB was 370-1110 GBq/µmol with a total synthesis time of ~40-min from EOB.