Blocking EP4 down-regulates tumor metabolism and synergizes with anti-PD-1 therapy to activate natural killer cells in a lung adenocarcinoma model.

Prostaglandin E2 (PGE2), a product of the cyclooxygenase (COX) pathway, is produced by tumors and surrounding stromal cells. It stimulates tumor progression, promotes angiogenesis and suppresses the anti-tumor response. Pharmacological inhibition of PGE2 synthesis has been shown to suppress tumor initiation and growth in vivo. In the current study, we demonstrated that the growth of the Ptgs2-deficient 3LL lung adenocarcinoma cell line was down-regulated in vivo through natural killer (NK) cell activation and a reduction in the population of polymorphonuclear leukocyte-myeloid-derived suppressor cells (PMN-MDSCs) and tumor-associated macrophages (TAMs). On the basis of these results, the therapeutic effect of ONO-AE3-208 (EP4i), an inhibitor of EP4 (a PGE2 receptor), combined with anti-PD-1 antibody was evaluated. EP4i, but not anti-PD-1 antibody, decreased tumor metabolism including glycolysis, fatty acid oxidation and oxidative phosphorylation. EP4i induced IFNγ production from only NK cells (not from T cells) and a shift from M2-like to M1-like macrophages in TAMs. These effects were further enhanced by anti-PD-1 antibody treatment. Although CD8 T-cell infiltration was increased, IFNγ production was not significantly altered, even with combination therapy. Tumor hypoxia was ameliorated by either EP4i or anti-PD-1 antibody treatment, which was further affected by the combination. Normalization of tumor vessels was significant only for the combination therapy. The results indicated a novel effect of EP4i for the metabolic reprogramming of tumors and revealed unique features of EP4i that can synergize with anti-PD-1 antibody to promote IFNγ production by NK cells, polarize TAMs into the M1 phenotype, and reduce hypoxia through normalization of the tumor vasculature.

Massive digital gene expression analysis reveals different predictive profiles for immune checkpoint inhibitor therapy between adenocarcinoma and squamous cell carcinoma of advanced lung cancer.

BACKGROUND: Immune checkpoint inhibitors prolong the survival of non-small cell lung cancer (NSCLC) patients. Although it has been acknowledged that there is some correlation between the efficacy of anti-programmed cell death-1 (PD-1) antibody therapy and immunohistochemical analysis, this technique is not yet considered foolproof for predicting a favorable outcome of PD-1 antibody therapy. We aimed to predict the efficacy of nivolumab based on a comprehensive analysis of RNA expression at the gene level in advanced NSCLC. METHODS: This was a retrospective study on patients with NSCLC who were administered nivolumab at the Kansai Medical University Hospital. To identify genes associated with response to anti-PD-1 antibodies, we grouped patients into responders (complete and partial response) and non-responders (stable and progressive disease) to nivolumab therapy. Significant genes were then identified for these groups using Welch's t-test. RESULTS: Among 42 analyzed cases (20 adenocarcinomas and 22 squamous cell carcinomas), enhanced expression of MAGE-A4, BBC3, and OTOA genes was observed in responders with adenocarcinoma, and enhanced expression of DAB2, HLA-DPB,1 and CDH2 genes was observed in responders with squamous cell carcinoma. CONCLUSIONS: This study predicted the efficacy of nivolumab based on a comprehensive analysis of mRNA expression at the gene level in advanced NSCLC. We also revealed different gene expression patterns as predictors of the effectiveness of anti PD-1 antibody therapy in adenocarcinoma and squamous cell carcinoma.

Dynamics and Mass Balance of Polycyclic Aromatic Hydrocarbons in and Around the Seto Inland Sea, Japan.

Polycyclic aromatic hydrocarbons (PAHs) were analyzed to elucidate the distribution, ecological risk, pathways, and fluxes of these pollutants in and around the Seto Inland Sea, an industrialized coastal region of Japan. High molecular weight PAHs (5-6 rings) were primarily found in regions close to the bay estuaries, and their proportions decreased at distances further from the estuaries (offshore areas), where low molecular weight PAHs (2-4 rings) were more ubiquitous. Screening-level risk assessments revealed that the PAHs found in the sediments should have no adverse effects on benthic communities. A mass balance for PAHs in the Seto Inland Sea, calculated based on data collected in the field and published literature findings, showed the PAH flux into the Seto Inland Sea from atmospheric deposition were ca. 6 times higher than that from riverine inflows. Comparison of the amount of the PAH mass flux between the Seto Inland Sea and the sea of the Europe and Asian countries indicated that the Seto Inland Sea is less polluted than the Gulf of Lion, the Mediterranean Sea, and the Bohai Sea, China and more polluted than the Yellow Sea. This paper is the first to determine the fluxes of PAHs in the coastal region of Japan.

Amlexanox enhances the antitumor effect of anti-PD-1 antibody.

Cancer immunotherapy, especially treatment with monoclonal antibodies (mAbs) that block programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) signaling, has attracted attention as a new therapeutic option for cancer. However, only a limited number of patients have responded to this treatment approach. In this study, we searched for compounds that enhance the efficacy of anti-PD-1 mAb using mixed lymphocyte reaction (MLR), which is a mixed culture system of the two key cells (dendritic and T cells) involved in tumor immunity. We found that amlexanox enhanced production of interferon (IFN)-γ, an indicator of T cell activation, by anti-PD-1 mAb. Amlexanox also induced PD-L1 expression in dendritic cells in MLR, whereas it did not stimulate interleukin-2 production by Jurkat T cells. These results suggest that amlexanox acts on dendritic cells, not T cells, in MLR. Furthermore, it enhanced the antitumor effect of the anti-PD-1 mAb in vivo in a mouse tumor-bearing model. The combination of amlexanox and anti-PD-1 mAb increased the expression of Ifng encoding IFN-γ, IFN-γ-related genes, Cd274 encoding PD-L1, and cytotoxic T cell-related genes in tumors. In conclusion, amlexanox stimulates the antitumor effect of anti-PD-1 mAb by acting on dendritic cells, which in turn activates cytotoxic T cells in tumors.

Singlet Oxygen Photogeneration in Coastal Seawater: Prospect of Large-Scale Modeling in Seawater Surface and Its Environmental Significance.

Chromophoric-dissolved organic matter (CDOM) acts as the precursor to singlet oxygen (1O2) in natural waters, while water acts as the main scavenger. In this study, we showed that 1O2 in coastal seawater can be successfully predicted from CDOM parameters. The 1O2 steady-state concentration [1O2]ss and photoformation rate (R1O2) varied by a factor of 6 across 13 sampling stations in the Seto Inland Sea, Japan, ranging from 1.2 to 8.2 × 10-14 M and 3.32 to 22.7 × 10-9 M s-1, respectively. Investigation of CDOM optical properties revealed that CDOM abundance measured as the absorption coefficient at 300 nm (a300) had the strongest correlation (r = 0.96, p < 0.001) with [1O2]ss, while parameters indicative of CDOM quality (e.g., spectral slope) did not influence [1O2]ss. A linear relationship between [1O2]ss and a300, normalized to a sunlight intensity of 0.91 kW/m2, was derived as [1O2]ss (10-14 M) = 2.12(a300) + 0.48. This was then used to predict [1O2]ss using a300 values from a subsequent, independent sampling exercise conducted 2 years after the first sampling. There was a good agreement (r = 0.93, p < 0.001) between the predicted values and the experimentally determined values based on a 95% prediction interval plot. Kinetic estimations using [1O2]ss suggest that 1O2 mediates the degradation of tetrabromobisphenol A in surface seawater (t1/2 = 0.63 days) while also contributing to the indirect photolysis of methyl mercury. The findings from this study suggest that large-scale modeling of 1O2 generation in surface seawater from CDOM parameters is possible with useful environmental significance for determining the fate of pollutants.

Evaluation of Levels, Sources and Health Hazards of Road-Dust Associated Toxic Metals in Jalalabad and Kabul Cities, Afghanistan.

This study was designed to investigate selected road-dust associated heavy metals, their relations with natural and anthropogenic sources, and potential human and environmental health risks. For this purpose, 42 and 36 road-dusts samples were collected from Jalalabad and Kabul cities (Afghanistan), respectively. The following elements were found in descending concentrations: Mn, Zn, Pb, Ni, Cu, Cr, Co, and Cd in Jalalabad; and Mn, Zn, Ni, Cu, Cr, Pb, Co, and Cd in Kabul. Except for Ni, all the elemental contents were less than the Canadian permissible limits in residential/parkland soils. Principle Component Analysis and enrichment of Cd, Cu, Ni, Pb, and Zn pointed to anthropogenic sources, whereas Co, Cr, and Mn indicated crustal inputs. Broadly, Cd monomial risk index ([Formula: see text]) was considerable; however, one site each in both cities showed high risk ([Formula: see text] ≥ 350). The potential ecological risk (RI) is mostly low; however, at some sites, the risk was considerable. Ingestion appeared to be the main exposure route (99%) for heavy metals and contributed > 90% to noncancerous (all residents), as well as 92% (children) and 75-89% (adults) cancerous risks. The noncancerous risks of all metals and their integrated risks for all residents were within acceptable levels. Moreover, potential cancer risks in children from Ni and Cr were slightly higher than the US-EPA safe levels but were within acceptable levels for adults. This study found higher risks to children and therefore recommends proper management and ways to control metals pollution load in these areas to decrease human health and RIs.

Spatio-temporal variations of selected pesticide residues in the Kurose River in Higashi-Hiroshima city, Japan.

This study investigated spatio-temporal variations of selected pesticide residues in the Kurose River in Higashi-Hiroshima city (Hiroshima Prefecture), Japan. Water samples were collected from the river at seven sites every month for 1 year (March 2016 to February 2017). Pesticide residues were extracted from the samples by a solid phase extraction using Sep Pack C18 cartridges. Once extracted, the samples were analyzed for cyanazine, simetryn, fenarimol, isoprothiolane, and diazinon using a reversed-phase high-performance liquid chromatography ultraviolet visible (HPLC-UV Vis) system. The limits of detection were 3.60, 4.10, 2.80, 6.50, and 7.30 ng L-1 for cyanazine, simetryn, fenarimol, isoprothiolane, and diazinon, respectively. Good recovery rates (88%-102%), and mean percent relative standard deviation range (1.00%-5.70%) (n = 6) were obtained with a spiking at 0.20 µg L-1. The maximum concentrations of 282, 391, 60, 1086, and 1194 ng L-1 were obtained for cyanazine, simetryn, isoprothiolane, fenarimol, and diazinon, respectively. Cyanazine was the most frequently detected pesticide (64% of the samples, n = 84), followed by simetryn (58%), and then diazinon (57%). The highest and lowest pesticide concentrations were measured during the periods May-June, and January-February, respectively. Principal component analysis revealed three principal components in which the pesticides were linked to dissolved organic matter and total suspended solids. The major water quality parameters (electrical conductivity, pH, Na+, K+, Mg2+, Ca2+, NH4+, NO3-, Cl-, SO42-, NO2-, and temperature) showed no clear trends for these pesticides. The presence of simetryn and isoprothiolane was largely attributed to rice paddy farms, whereas diazinon was associated mostly with vegetable farms and orchards. The diazinon and isoprothiolane patterns were consistent with their use of controlling insects and fungi in the prefecture. The maximum diazinon concentration detected was higher than the human safe level specified by the European Union (100 ng L-1) in Council Directive 98/83/EC. This is of concern because of the bioconcentration potential of these residues in fish and other marine animals consumed by humans.

Novel Fluorometric Method for the Determination of Production Rate and Steady-State Concentration of Photochemically Generated Superoxide Radical in Seawater Using 3',6'-(Diphenylphosphinyl)fluorescein.

Superoxide radical (O2(•-)) is an important reactive oxygen species in seawater. Measurements of its production rates and steady-state concentrations generated by photochemical processes have been a Herculean task over the years. In this study, a probe - 3'6'-(diphenylphosphinyl)fluorescein (PF-1) - was used to trap photochemically generated O2(•-) in seawater, thereby yielding fluorescein. The fluorescein produced was measured by an isocratic fluorescence HPLC at excitation/emission wavelengths of 490/513 nm, respectively. The reaction rate constant of PF-1 with O2(•-) (kPF-1) was pH-dependent: (3.2-23.5) × 10(7) M(-1) s(-1) at pHTOT 7.65-8.50. By applying appropriate equations, both the production rate and the steady-state concentration of O2(•-) generated by photochemical reactions in the seawater were quantified. Under the optimized experimental conditions, fluorescein standards (3-50 nM) exhibited linearity in the seawater by HPLC. The photoformation of fluorescein, due to the reaction of PF-1 with the O2(•-) photochemically produced in the seawater, was linear within the 20 min irradiation. The detection limit of the fluorescein photoformation rate was 0.03 pM s(-1), defined as 3σ of the lowest standard fluorescein concentration per 20 min irradiation. Using this value, the yield of fluorescein, and the fraction of O2(•-) that reacted with PF-1 in the seawater, the detection limit of the O2(•-) photoformation rate was 1.78 pM s(-1). Superoxide measurements using the proposed method were relatively unaffected by the potential interfering species in seawater. Application of the proposed method to ten (10) seawater samples from the Seto Inland Sea, Japan, resulted in measured O2(•-) photoformation rates of 3.1-8.5 nM s(-1), with steady-state concentrations ranging (0.06-0.3) × 10(-10) M. The method is simple, requires no technical sample preparation, and can be used to analyze a large number of samples.

Highly sensitive detection of hepatitis B virus surface antigen by use of a semiautomated immune complex transfer chemiluminescence enzyme immunoassay.

The performance of hepatitis B surface antigen (HBsAg) screening assays is continuously improved to reduce the risk of transfusion-associated hepatitis B. In this study, a semiautomated immune complex transfer chemiluminescence enzyme immunoassay (ICT-CLEIA) for the detection of HBsAg, which is as sensitive as hepatitis B virus (HBV) DNA PCR, was developed; the ICT-CLEIA assay performance was compared with the performance of the Architect HBsAg QT assay and HBV DNA PCR. The specificities in the initial assay and after retesting were 99.50% (1,988/1,998 samples) and 99.95% (1,997/1,998 samples), respectively. The analytical detection limit was determined to be 0.2 mIU/ml using the 2nd International WHO HBsAg standard, and the cutoff value (0.5 mIU/ml) of the ICT-CLEIA assay was 8.0 standard deviations (SD) above the mean of the HBsAg-negative specimens. The ICT-CLEIA assay could detect HBsAg even in the presence of anti-HBs antibodies and demonstrated a 23.6-day-shorter window period using commercially available HBsAg seroconversion panels than the Architect HBsAg QT assay. Furthermore, the monitoring of the viral kinetics by the ICT-CLEIA assay and the HBV DNA PCR produced very similarly shaped curves during both the HBsAg seroconversion and reverse seroconversion periods. Therefore, the ICT-CLEIA assay may be useful not only for an earlier detection of HBV reactivation but also for the monitoring of hepatitis B patients.

Phenol formation in gamma radiolysis of aqueous benzene solution with sodium hypochlorite.

Phenol formation by gamma radiolysis of an aqueous benzene solution containing sodium hypochlorite is reported. The phenol formation in a benzene solution containing sodium hypochlorite irradiated with (60)Co γ-rays is about six times higher than that without sodium hypochlorite. Ten micromolar sodium hypochlorite enhanced the formation of phenol up to a total dose of 6 Gy. Above 6 Gy in solutions containing sodium hypochlorite, the rate of phenol yield sharply decreased and was essentially the same as that without sodium hypochlorite. The yield of phenol with sodium hypochlorite is 0.89 µmol J(-1) and is larger than the sum of yield for the radicals and reactive oxygen species by water radiolysis such as •OH, e(-), H, H2, and H2O2. The formation of phenol with sodium hypochlorite was reduced by NaCl. Results suggest that the radiolytic formation of phenol in a benzene solution with sodium hypochlorite relates to the reaction process involving chlorine atoms. Sodium hypochlorite can be applied as a sensitizer for a benzene chemical dosimetry system. The lower limit of dose detection calculated from the detection limit of phenol and the G value of phenol was estimated to be 1 × 10(-3) Gy.

Photodegradation of the insecticide fipronil in aquatic environments: photo-dechlorination processes and products.

Fipronil (FIP) is a phenylpyrazole insecticide that, along with neonicotinoid insecticides, is regularly used worldwide. Photodegradation of FIP in aqueous systems is thought mainly to involve the reaction of desulfinylation to give fipronil desulfinyl (FIP-desulfinyl); however, little is known about further degradation reactions. We investigated FIP photodegradation by analyzing photodegradation products by liquid chromatography and liquid chromatography high-resolution tandem mass spectrometry using an Orbitrap instrument. A wide range of products, including dechlorinated compounds, was detected, and the structures were identified. FIP-desulfinyl has previously been found to be an important and persistent FIP photodegradation product; however, we also found that FIP-desulfinyl was photochemically decomposed to a didechlorinated product via a monodechlorinated product. The main photodegradation pathway was probably similar to that of ethiprole, which has a similar skeleton. The photodegradation rate constant was 22.6 times lower for FIP-desulfinyl (0.00372 min-1) than FIP (0.0839 min-1). The photodegradation rate constant was lower for the newly found didechlorinated product (0.001 min-1 or below) than FIP-desulfinyl, suggesting that the product is persistent in aquatic environments and could be an important indicator of long-term FIP contamination.

Distribution and phylogeny of mercury methylation, demethylation, and reduction genes in the Seto Inland Sea of Japan.

Mercury (Hg) adversely affects human and environmental health. To evaluate the mercury (Hg) speciation (methylation, demethylation, and reduction) of microorganisms in coastal seawater, we analyzed the microbial functional gene sets involved in Hg methylation (hgcA and hgcB), demethylation (merB), and reduction (merA) using a metagenomic approach in the eastern and western parts (the Kii and Bungo channels, respectively) of the Seto Inland Sea (SIS) of Japan. We determined the concentration of dissolved total mercury (dTHg) and methylated mercury (dMeHg) in seawater. The metagenomic analysis detected hgcAB, merA, and merB in both channels, whereas the phylogenies of these genes differed between them. A correlation between Hg concentration (both dTHg and dMeHg) and the relative abundance of each gene was not observed. Our data suggests that microbial Hg methylation and demethylation could occur in the SIS and there could be a distinct microbial Hg speciation process between the Kii and Bungo channels.

Mechanisms of hydrogen sulfide removal with steel making slag.

In the present study, we experimentally investigated the removal of hydrogen sulfide using steel-making slag (SMS) and clarified the mechanism of hydrogen sulfide removal with the SMS. The results proved that SMS is able to remove hydrogen sulfide dissolved in water, and the maximum removal amount of hydrogen sulfide per unit weight of the SMS for 8 days was estimated to be 37.5 mg S/g. The removal processes of hydrogen sulfide were not only adsorption onto the SMS, but oxidation and precipitation as sulfur. The chemical forms of sulfide adsorbed onto the SMS were estimated to be sulfur and manganese sulfide in the ratio of 81% and 19%, respectively. It is demonstrated here that the SMS is a promising material to remediate organically enriched coastal sediments in terms of removal of hydrogen sulfide. Furthermore, using SMS is expected to contribute to development of a recycling-oriented society.

Migration of terephthalate from scraps of poly(ethylene terephthalate) (PET) in water and artificial seawater.

We report the migration of terephthalate and some low molecular weight organic compounds from poly(ethylene terephthalate) (PET) scraps in Milli-Q water and artificial seawater (ASW). The photochemical processes and the subsequent dark reactions were investigated using PET scraps obtained from postconsumer bottles of commercial non­carbonated mineral water. Concentrations of terephthalate exponentially increased with irradiation time, reaching approximately 6-8 µmol L-1 in ASW after 80 h irradiation. The photochemical migrations of compounds related to terephthalate were also observed. Concentrations of terephthalate and related compounds reached higher concentrations in ASW than in Milli-Q water. After 80 h irradiation, two dark experiments were conducted: one on the solutions after irradiation without PET scraps, and the other on photochemically damaged PET scraps. In ASW in the dark without PET scraps, the terephthalate concentration increased, and concentrations of other compounds related to terephthalate also changed. The results suggested that terephthalate was generated by hydrolytic reaction in dark ASW from the scission products of PET which were generated during the irradiation of PET scraps. Photochemically damaged PET scraps released terephthalate and related compounds in the dark. The half-life of the photo-irradiated PET scraps in the dark is approximately 205 years. Our results show that PET bottles in marine environments can continuously release terephthalate and other low molecular weight organic compounds during the day at the sunny surface, at the dark ocean floor, and during the night.

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

We assessed the contamination, dynamics, and health risks of the pesticides cyanazine, simetryn, fenarimol, isoprothiolane, diazinon, irgarol, fenitrothion, and diuron in marine samples (seawater, sediments, plankton, fish, and other edible organisms) at various locations in the Seto Inland Sea in Japan in 2016 and 2017. Pesticide concentrations were highest at sampling sites close to the coastline, and mean concentrations in seawater were slightly higher in surface water than in bottom water. All eight pesticides were detected in plankton. Diazinon concentrations (77-387 ng/g dw) were highest in sediments and cyanazine was the most frequently detected pesticide (88%, n = 17) in sediments. Only cyanazine (2.7-41.9 ng/g dw), simetryn (1.0-34.3 ng/g dw), and diazinon (6.3-308.8 ng/g dw) were detected in fish and other edible marine organisms. Based on the calculated bioconcentration factor, the results showed that plankton, fish, and marine animals bioaccumulated pesticides. The highest hazard quotients were calculated for diazinon in red seabream and greenling, indicating a possible risk to consumers. It is, therefore, imperative to promote strict implementation of pollution control, integrated pest management practices, and policy formulation on pesticides. Usage of diazinon must be controlled and monitored to ensure large residues do not reach aquatic ecosystems and marine coastlines.

Optical properties of dissolved organic matter in Japanese rivers and contributions to photoformation of reactive oxygen species.

The optical properties of dissolved organic matter (DOM) from five rivers (Kokubu, Kurose, Ohta, Yamato, and Yodo) in Japan were investigated and contributions of DOM to photoformation of three reactive oxygen species (ROS) (hydroxyl radicals (OH), nitric oxide radicals (NO), and singlet oxygen (1O2)) were assessed. The lowest and highest mean dissolved organic carbon concentrations were for the Ohta River (0.95 (mg C) L-1) and Yamato River (2.85 (mg C) L-1), respectively, and the concentrations correlated with some optical parameters. Absorption ratios (e.g., the E2:E3 and A280/A350 ratios) and the spectral slope S275-295 indicated that DOM from the Yodo and Kokubu rivers had the lowest and highest molecular weights, respectively. PARAFAC models and DOM excitation-emission matrices were used to assess the sources and fates of DOM in the rivers. The PARAFAC model indicated that the main types of fluorescent DOM in the rivers were terrestrial humic-like (TH-L) and tryptophan-like (TP-L) substances. The Kokubu River contained other compounds such as fluorescent whitening agents, autochthonous humic-like substances, and extracellular polymeric substances. Statistically significant relationships between the dissolved organic carbon and TH-L, TP-L, and extracellular polymeric substance concentrations suggested that TH-L, TP-L, and extracellular polymeric substances are important contributors to total DOM in the rivers. TH-L and TP-L substances strongly contribute to ROS photoformation, but TH-L substances play roles in both ROS generation and scavenging. Comprehensive models for estimating the photoformation rates of different ROS (in M s-1) were established by integrating the contributions of the relevant major and minor sources. Examples are ROH (10-12) = 21.0 [NO2-]_µM + 0.460 [TH-L]_QSU + 10.9, RNO (10-12) = 67.9 [NO2-]_µM + 35.2 [a300]_m-1 - 2.51 [TH-L]_QSU - 0.765 [TP-L]_QSU - 8.14, and R1O2 (10-9) = 3.81 [a300]_m-1 - 0.101 [TP-L]_QSU + 11.1.

Heavy metals in urban dusts from Alexandria and Kafr El-Sheikh, Egypt: implications for human health.

A total of 23 road-dust and 9 house-dust samples were collected from Alexandria and Kafr El-Sheikh cities, Egypt in 2016 to investigate heavy metal (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) contamination, spatial distribution, sources, and health risks. The mean concentrations (mg kg-1) of Cd (road-dust (RD) = 0.33, house-dust (HD) = 0.77), Cu (RD = 80, HD = 141), Pb (RD = 70, HD = 260), and Zn (RD = 169, HD = 771) in Alexandria and Zn (RD = 192, HD = 257) in Kafr El-Sheikh were higher than corresponding background (background refers to generic earth crust shale values given in the literature) levels. Whereas average concentrations (mg kg-1) of Co, Cr, Mn, and Ni (Alexandria: RD = 2.7, 24.3, 251, 14.4; HD = 3.2, 29.2, 237, 25.1 and Kafr El-Sheikh: RD = 6.6, 31.9, 343, 20.2; HD = 8.6, 33.4, 438, 23.2) in both cities were much lower than their background values. Spatially, for most heavy metals, the high concentrations were observed in areas characterized with increased anthropogenic activities, heavy traffic, and high population density. Contamination indices revealed moderate contamination (Cd and Cu) to high contamination (Pb: only house-dust from Alexandria), which posed low (most metals) to moderate ecological risk (Cd and Pb). Correlation analysis and factor analysis classified the studied metals in two groups as: natural input (Co, Cr, Mn, Ni, and Fe) and anthropogenic sources (Cd, Cu, Pb, and Zn). The noncancerous risks posed by studied metals ranged from 0.0001 (Cd) to 0.15 (Pb) and were insignificant. The cancerous risk of Pb (1.4 × 10-4) for children on exposure to house-dust form Alexandria exceeded the guideline values and was considered unacceptable, whereas the cancerous risks of other studied metals were acceptable for both subpopulations. The results of health risk revealed that children are facing higher risk than adults.

Photochemical production and consumption mechanisms of nitric oxide in seawater.

Nitric oxide (NO•) is an active odd-nitrogen species that plays a critical role in determining the levels of ozone (O3) and other nitrogen species in the troposphere. Here, we provide experimental evidence for photochemical formation of NO• in seawater. Photoproduction rates and overall scavenging rate constants were measured by irradiation of surface seawater samples collected from the Seto Inland Sea, Japan. Photoproduction rates of NO• ranged from 8.7 × 10⁻¹² M s⁻¹ to 38.8 × 10⁻¹² M s⁻¹ and scavenging rate constants were 0.05-0.33 s⁻¹. The steady state concentrations of NO• in seawater, which were calculated from the photoproduction rates and scavenging rate constants were in the range 2.4-32 × 10⁻¹¹ M. Estimation from the scavenging rate constant showed that the NO• lifetime in seawater was a few seconds. Our results indicate that nitrite photolysis plays a crucial role in the formation of NO•, even though we cannot exclude minor contributions from other sources. Analysis of filtered and unfiltered seawater samples showed no significant difference in NO• photoformation rates, which suggests a negligible contribution of NO• produced by photobiological processes. Using an estimated value of the Henry's law constant (kH ≈ 0.0019 M atm⁻¹), a supersaturation of surface seawater of 2 to 3 orders of magnitude was estimated. On the basis of the average values of the surface seawater concentration and the atmospheric NO• concentration, a sea-to-air NO• flux was estimated.

A fluorescence method for the determination of photochemically generated peroxynitrite in seawater.

Peroxynitrite (ONOO-) is a reactive species which can degrade several classes of organic compounds via both oxidation and nitration reactions. Despite the fact that the photochemical precursors; superoxide (O2·-) and nitric oxide (NO·) radicals, have been measured in seawater under simulated solar radiation, there is no published report on actual measurements of photoformed peroxynitrite in seawater. Hence, this attempt was made to develop a fluorescence method by using coumarin boronic acid (CBA) as a chemical probe. CBA reacts with ONOO- to yield fluorescent 7-hydroxycoumarin (COH). COH was determined by reversed‒phase isocratic HPLC with fluorescence detection at excitation/emission wavelengths of 332/475 nm, respectively. COH standards calibration is linear in range of (0.25-100) × 10-9 M. The detection limit, defined as 3σ of the lowest ONOO- photo-formation rates obtained within 10- min irradiation, was 1.66 × 10-12 M s-1. This method is very precise as coefficient of variation for triplicate measurements of COH photo-formation rates was a maximum of 0.052. Experimental procedures were optimized to handle potential interference by hypochlorite, and the method was applied to measure ONOO- in 13 surface seawater samples obtained from the Seto Inland Sea, Japan. ONOO- photo-formation rates, steady‒state concentrations and lifetimes were determined to be (0.06-5.13) × 10-9 M s-1, (0.98-6.11) × 10-11 M and (0.01-0.16) s, respectively.

Factors controlling the degradation of hydrogen peroxide in river water, and the role of riverbed sand.

Diurnal changes of H2O2 in river water during mid-summer were investigated. H2O2 in river water increased with the increase in intensity of solar radiation in the morning, and reached a maximum at 14:00, although solar radiation reached a maximum around 12:00. In the afternoon, a gradual decrease in H2O2 was observed, and H2O2 reached a minimum just before sunrise. Degradation rate constants determined using unfiltered river water samples were 0.081-0.161 h-1, corresponding to a half-life of 4.3-8.5 h. We simulated diurnal changes in H2O2 using a simple formation, accumulation, and degradation model for static water using formation and degradation rate constants. The results of the modeling suggested that in situ degradation rate constants in rivers could be faster than those determined for unfiltered river water samples. Experiments using river sand indicated that riverbed sand could play an important role in H2O2 decay in rivers. We discussed the decomposition process of H2O2 in rivers.