Occurrence of selected pharmaceuticals in river water in Japan and assessment of their environmental risk.

The existence of pharmaceuticals in the water environment is thought to be a potential problem for aquatic organisms. In this study, we conducted a nationwide survey to clarify the occurrence of 24 selected pharmaceuticals in major Japanese rivers and evaluated their environmental risk to aquatic organisms. We found a total of 22 substances in river waters at concentrations from several nanograms per liter to several micrograms per liter. We found the highest, which was 2.4 µg/L of caffeine, followed by 1.5 µg/L of crotamiton and 1.4 µg/L of sulpiride. We conducted an environmental risk assessment of the 22 pharmaceuticals detected in river water, for which predicted no-effect concentration (PNEC) values for crustacea and algae had been obtained. The measured environmental concentration/PNEC values of four substances, caffeine, carbamazepine, clarithromycin, and ketoprofen, exceeded 0.1 with the maximum value of 9.0 for clarithromycin. As clarithromycin exhibits a high environmental risk to aquatic organisms, particular attention is required.

Field survey of pollutants discharged from different types of residential area in the Yamuna River Basin, India.

The Ganges River, one of the most heavily populated and urbanized river basins in Asia, is polluted by increasing wastewater influent and water-borne diseases are caused in the metropolitan area. This study focused on the Yamuna River, a major tributary of the Ganges. We determined the pollutant load per unit of urban area classified by the income of the residents to help design an appropriate sewerage system. In addition, a simple method of estimating runoff pollutant load was examined using data on pollutant load per unit and runoff coefficient.

Extraction of nonylphenol and nonylphenol ethoxylates from sewage sludge using the pressurized fluid extraction method and the supercritical fluid extraction method.

To establish endocrine disruptor (ED) monitoring and control methods for wastewater treatment plants, it is necessary to clarify the fate of EDs in each process of a sludge treatment system. However, the ED analysis method for sewage sludge containing large quantities of organic matters has not been established. So, a highly reliable analytical method that accurately measures EDs in sewage sludge must be developed. This paper reports on the results of applying the Pressurized Fluid Extraction (PFE) method and the Supercritical Fluid Extraction (SFE) method to the extraction of nonylphenol (NP) and nonylphenol ethoxylate (NPnEO) existence of which has often been confirmed in past surveys. To clarify the quantity of EDs in sludge specimens and the fate of EDs in the sludge treatment process, appropriate analytical methods should be developed. In this study, the PFE method and the SFE method were tested to extract NP and NPnEO from sewage sludge specimens. The findings indicate that the extraction from dried sludge by the PFE method can extract a larger quantity of NP and NPnEO in a shorter time than the widely used heating reflux method. Regarding the extraction by the SFE method, conditions for extraction of NP and NPnEO at a higher rate than the heating reflux method were not found.

Heat island mitigation using water retentive pavement sprinkled with reclaimed wastewater.

In Japan, reclaimed wastewater has been recycled widely for non-potable urban applications and it is to be used for sprinkling roads to mitigate heat island in urban areas. To assess the heat island mitigation effects of the sprinkling reclaimed wastewater on water retentive pavement, we carried out a survey at Shiodome-District, Tokyo. The temperatures of air and roads, humidity, and WBGT (Wet-bulb globe temperature) were measured and heat flux was estimated to compare the condition of the areas with/without sprinkling. The following results were obtained. 1) Sprinkling reclaimed wastewater decreased the road surface temperature by 8 degrees during the daytime and by 3 degrees at night: temperatures equal to those on planting zones. Nevertheless sprinkling was done only in the daytime, the temperature decrease effect was not only obtained during the daytime: it continued through the night, due to the water retentive pavement. 2) Sprinkling reclaimed wastewater reduced the amount of sensible heat flux and increased that of latent heat flux. These results suggest that sprinkling reclaimed wastewater on water retentive pavement can effectively mitigate the heat island phenomenon.

Fate of nonylphenol and 17beta-estradiol contained in composted sewage sludge after land application.

Many environmental problems caused by endocrine disrupters (EDs) have been reported. Because little is known about the fate of EDs accumulated in sewage sludge, we carried out a study to clarify the fate of EDs in composted sludge after its application to soil. Nonylphenol (NP) and 17beta-estradiol (E2) were measured for leachate and soil. High concentrations of NP and E2 were detected in the leachate at the early stage, but they decreased rapidly. Also, the high contents of NP and E2 in soil decreased significantly within 300 days. Because the decrease of NP and E2 in the soil was much larger than that of NP and E2 in the leachate, there must have been a physicochemical or biological decomposition mechanism in the soil layer. We also tried to clarify the transfer of NPs to plants from compost. In the experimental conditions of this study, the transfer of NPs to plants from compost was not observed.

Management of chemical substances in a water environment communicating among stakeholders.

With the implementation of Pollutant Discharge and Transfer Register (PRTR) in fiscal 2003, information on the quantity of chemical substances discharged annually from each business is released in Japan. Such information is expected to lead to heightened public concern about the presence of chemical substances in environment. This paper showed the effects of using PRTR data for managing chemical risk in a water environment communicating among stakeholders based on the results of the case study held in Japan. First, it was possible to identify the discharge sources using PRTR and related data for the chemicals such as Zn and NPs which were shown that they had relatively high risks based on the survey in a model area. Second, it was suggested that the chemical risk communication among the stakeholders including people using PRTR and related data would be a good way to encourage environmental activities of businesses, and to build a good relationship among stakeholders including people who have anxiety about chemical risk.

Relationship of nutrient and residual chlorine concentration in treated wastewater with periphytic algae grown in a stream receiving treated wastewater.

The ratio of the amount of treated wastewater to river water is increasing in urban areas due to the spread of sewage systems. Treated wastewater is also sometimes extensively used to create streams and other water environments in urban areas. Rivers, streams and other water environments provide valuable habitats for all kinds of aquatic species, but the relationship between such aquatic species and the quality of treated wastewater they inhabit is not clearly understood. This study was carried out to clarify the effect of the water quality of treated wastewater such as nutrients and residual chlorine on periphytic algae grown in a stream receiving treated wastewater using laboratory-scale experimental channels. The following results were obtained. (1) When the range of phosphate (PO4-P) concentration was 0.04 to 0.09 mg/L, the higher the PO4-P concentration, the higher the biomass of periphytic algae and the more dominant the Chlorophyceae. (2) When the range of total residual chlorine (TRC) concentration was 0.07 to 5.8 mg/L, the higher the TRC concentration, the lower the biomass of periphytic algae. When the range of TRC concentration was 0.93 to 5.8 mg/L, this tendency was more pronounced.

Comprehensive management of chemical substances in a water environment using PRTR data.

The annual quantity of discharge of 354 chemical substances from each business has been released based on PRTR law in Japan since 2003. MLIT and municipalities should be responsible for managing chemical risk in public water bodies through communication with stakeholders, such as the public and businesses. However, it is economically difficult to measure the discharge loads and behaviour of all chemical substances and reveal which substances should be particularly managed in each basin. NILIM has begun studies on survey methods to understand the discharge and behaviour of chemical substances in each basin based on PRTR information, and to conduct risk management of chemical substances in cooperation with stakeholders such as the public and businesses. We selected 30 chemical substances, such as those listed or nominated in environmental criteria and endocrine disrupters, collected the volume of discharge of chemical substances from public and industrial wastewater treatment plants using PRTR information, and surveyed their behaviour in the river in the model area. As a result, the discharge of 12 chemical substances were reported in PRTR, and 17 substances were detected in the river. Some inorganic compounds were detected intensively near discharge sites reported in the PRTR. But some organic compounds and endocrine disrupters, such as oestrone, were detected although their discharge was not reported in the model area in the PRTR. It suggested that PRTR information is useful to identify hot spots, but further study is needed to understand the discharge of chemical substances from small businesses, farmland and houses.

Fate of nonylphenol polyethoxylates and nonylphenoxy acetic acids in an anaerobic digestion process for sewage sludge treatment.

Many environmental problems caused by endocrine disruptors (EDs) have been reported. It is reported that EDs flow into sewage treatment plants, and it has been pointed out that these may be shifted from the wastewater treatment process to the sludge treatment process. Little is known about the fate of EDs accumulated in sewage sludge, so we carried out a study to clarify the fate of EDs in sewage sludge treatment processes, especially in an anaerobic digestion process. In this study, nonylphenol (NP) was selected as a target ED. Nonylphenol ethoxylates (NPnEO) or nonylphenoxy acetic acids (NPnEC), which were the precursor of NP, were added to an anaerobic digestion process, and mass balance was investigated. The following results were obtained from the anaerobic digestion experiments. (1) NP1EO was injected to an anaerobic digestion testing apparatus that was operated at a retention time of approximately 28 d and a temperature of 35 degrees C with thickened sludge sampled from an actual wastewater treatment plant. Approximately 40% of the injected NP1EO was converted to NP. (2) NP1EC was injected to an anaerobic digestion testing apparatus with thickened sludge. As a result, almost all injected NP1EC was converted to NP. When NP2EC was injected, NP2EC was not converted to NP until the 20th day.

Cloning and characterization of the rat TIS11 gene.

The TIS11 gene is an immediate early gene that is induced rapidly and transiently by phorbol 12-myristate 13-acetate and various growth factors. To study transcriptional regulation of the gene, a genomic clone of rat TIS11 was isolated, and the organization of exon-intron structure and transcriptional initiation site were determined. The rat TIS11 gene consisted of 2 exons spanning approximately 2.5 kb. Several canonical sequences for binding of transcriptional factors were found in the 5'-flanking region. The 5.3 kb of the 5'-flanking region fused to a luciferase reporter gene showed promoter activity when introduced into rat pheochromocytoma PC12 cells. Analyses with serial 5'-deletion mutants suggested that the major positive regulatory region is located at the region of -241 to -76, and that the minimum promoter region is within the 76-bp upstream of the transcriptional initiation site. Gel mobility shift assays revealed that PC12 cell nuclear proteins specifically bind to the major positive regulatory region of the TIS11 gene. The identified nuclear protein components may act as the positive trans-acting factors in the basal expression of the TIS11 gene in PC12 cells.

Microvascular blood is distributed more to venules than to arterioles in patients with Buerger's disease. Observation of bulbar conjunctiva by intravital microscope system.

BACKGROUND: It has been a matter of controversy whether abnormalities of organs other than extremities may be a clinical manifestation of Buerger's disease (thromboangiitis obliterans; TAO). In the present investigation, our aim was to quantitatively characterise the configuration of microvascular networks in bulbar conjunctiva, which is not affected apparently, in patients with thromboangiitis obliterans. METHODS: Nine men with thromboangiitis obliterans attended our hospital and nine male volunteers as normal controls were enrolled in this study. We observed and analysed the configuration of the network of a bulbar conjunctiva by use of intravital microscope system with computer assisted image processing functions. Microvessel density was defined as a summation of vessel length in a ROI area and tortuosity was evaluated by a ratio of vessel length to direct distance of both terminals. RESULTS: In the microcirculation of bulbar conjunctiva in thromboangiitis obliterans, arteriole diameter was significantly decreased and density of venules was significantly increased. Increased venular density was mainly explained by increased tortuosity of venules. CONCLUSIONS: Consequently, microvascular blood was distributed more to venules than to arterioles in patients with thromboangiitis obliterans. Venule/arteriole ratios of diameter, tortuosity and microvessel density may be useful parameters to characterize the configuration of microvascular networks in thromboangiitis obliterans.

Macro- and microcirculatory effects of tetramethylpyrazine and extract of Charthamus tinctorius L. in rabbit mesentery.

Effects of tetramethylpyrazine (TMP) and that of extracts of Charthamus tinctorius L. (CTL) on the macro- and microcirculation in rabbit mesentery were studied. The intestinal arterial blood flow (Qa) was measured using an electromagnetic flowmeter, together with the arterial blood pressure (Pa). The inner diameter and red cell velocity in single arteriole in the mesentery were measured by a video-image technique and a dual-slit photometric method, respectively. Using the measured diameter and red cell velocity, the arteriolar blood flow (Qm) was calculated. Both the Qa and Qm decreased when Pa was decreased after the intravenous administration of TMP, CTL, Nicardipine, Phentolamine and acetylcholine (Ach). Changes in Qa and Qm with changes in Pa were analyzed, and it was found that (i) both the Pa-Qa and Pa-Qm curves, during the administration of TMP or CTL, show different patterns from those during the administration of Nicardipine or Phentolamine; (ii) the Pa-Qa and Pa-Qm curves after the administration of TMP or CTL show similar patterns with those after the administration of Ach.

[In vivo visualization of renal microcirculation using hydronephrotic rat kidney].

A number of neurohumoral factors participate in the regulation of renal hemodynamics. Several methods have been developed to study directly the regulation of renal microcirculation. Here, we introduce an in vivo method to visualize renal microcirculation by using hydronephrotic rat kidney, a unique method originally developed by Steinhausen et al. More than 10 weeks after unilateral ureteral ligation in rats, the renal parenchyme becomes thinner and suitable for transillumination. After anesthesia, the hydronephrotic kidney was split at the greater curvature with a thermal cautery and then fixed in a water chamber containing Kreb's solution. The renal tissue was transilluminated and microscopically visualized using water immersion objectives. Renal microvessels including arcuate and interlobular arteries, afferent and efferent arterioles and glomerular capillaries could be easily observed on a display monitor at a final magnification of 2,700 times. Topical application of angiotensin II elicited constriction of the interlobular artery and afferent and efferent arterioles dose-dependently. Thus, this preparation is a unique model allowing visualization of the whole renal vascular tree in vivo.

Propagation properties of vasomotion at terminal arterioles and precapillaries in the rabbit mesentery.

A vasomotion activity in the mesentery of anesthetized rabbits were studied by simultaneous measurements of inside diameters at multiple sites in arterioles, precapillaries and their bifurcations. A frame-by-frame diameter determination technique was used with a microcomputer-assisted laser video disk recorder and video-image analysis system. Simultaneous intensity profiles across microvessels were continuously obtained. Applying an automatic wall surface tracer and a graphic editor to construct temporal sequences of intensity profiles, we obtained digitized data of inside diameters of microvessels, and implemented cross-correlation analysis between data sets to calculate phase differences of vasomotion at separated sites. The present analysis of the propagation of vasomotion showed that the vasomotion originated from the orifice of precapillaries at the bifurcations, spreading downstream in the precapillary. The vasomotion wave spreads both upstream and downstream along single arterioles from various origins of the vasomotion activity. The propagation velocity of vasomotion was 0.17 +/- 0.03 mm/sec (n = 19), and it became significantly slower through the branching points than along the arterioles. It is suggested that the vasomotion in terminal arterioles and precapillaries may spread through some mechanical transmission factors.

Vascular resistance of arterioles with nonuniform diameters.

Arterioles in various vascular beds have been often observed to have nonuniform diameters along the vessel axis. The effect of nonuniformity of arteriolar diameter on vascular resistance was investigated using a theoretical model of blood flow in arterioles. Viscous flow of a Newtonian fluid in tubes with periodically changing diameters along the tube axis was analyzed by a finite element method based on the Stokes equations. Vascular resistance for the nonuniform tube was computed over a spatial period of the variation in diameter and was compared to resistance for a uniform tube with a constant diameter equal to the mean diameter of the nonuniform tube. In all cases, resistance for a tube with a nonuniform diameter was larger than resistance for a uniform tube with a diameter equal to the mean diameter of the nonuniform tube. Increases in the amplitude of the variation in diameter resulted in a rapid increase in resistance when the period of the variation remained constant. On the other hand, as the period of the diameter variation increased when amplitude remained constant, resistance decreased and approached the values obtained under the assumption of a Poiseuille flow at each cross section of the tube in the limit of an infinite period. Our theoretical model was applied to our previous in vivo studies of vessel diameter nonuniformity for rabbit mesentery arterioles in a contracted state. It was shown that vascular resistance calculated by our model was 2 to 11% higher than resistance obtained for a uniform tube with a diameter equal to the mean diameter of the arteriole.

The control of macro- and microcirculation in the mesentery and intestine.

The total blood flow and microvessel blood flow in the mesentery and the intestine anesthetized rabbits were measured while systemic hemodynamics were altered by baroreceptor stimulation and by noradrenaline injection. The percent change of the total peripheral resistance (Rt) was greater than the change in resistance in the intestinal vascular bed (Ri) during baroreceptor stimulation. The capillary blood flow in the intestinal muscle layer (Qs) and villus (Qv) increased slightly while the intestinal arterial flow (Qa) and the arteriolar flow in the mesentery (Qm) decreased. These data indicated that Qs and Qv were not controlled by the sympathetic vasoconstrictor system. The percent change of Ri was greater than that of Rt during noradrenaline injection. Noradrenaline decreased Qa, but Qs and Qv were decreased while Qm was increased. These results show that noradrenaline stimulates alpha-adrenoceptors on small arteries and arterioles in the intestine. The observed Qm changed was in response to the systemic hemodynamic change caused either by the baroreceptor input or by noradrenaline.

Heterogeneous response in hepatic sinusoidal blood flow to acute ethanol ingestion in rats.

The effect of ethanol ingestion on the hepatic sinusoidal blood flow velocity was investigated in rats, using in vivo microscopy system with dual-slit photometric method. In steady state, before ethanol ingestion, the hepatic sinusoidal blood flow velocity determined at 18 points ranged widely from 54 to 245 micron/sec and the mean value was 144 +/- 13 micron/sec. After acute ethanol ingestion (1g/kg), the averaged sinusoidal blood flow velocity increased significantly to 197 +/- 17 micron/sec, while the width of sinusoids showed no apparent change. In some sinusoids, however, the blood flow velocity did not increase. Thus, the heterogeneous response in hepatic sinusoidal blood flow to the ethanol metabolism might induce the development of an imbalance between delivery and demand of oxygen to hepatocytes around those sinusoids after acute ethanol ingestion.

Thrombus formation by the application of thrombin to the outer surface of mouse mesenteric vein: comparison with the application of ADP.

Mesenteries of mice under anesthesia were stretched over an inverted microscope. A micropipette filled with solution containing various concentrations of ADP or thrombin was brought into contact with the outside of a mesenteric vein by micromanipulation, and then poured over the outer surface of the vein. Morphological characteristics of the thrombi and the time needed for thrombus formation were examined. Application of either thrombin or ADP to the adventitia of mesenteric veins caused thrombus formation. Although thrombi by application of ADP seemed to be anchored by direct adhesion of platelets to the exposed subendothelium, thrombi by application of thrombin seemed to be anchored by deposited fibrin.

High susceptibility to ADP-induced thrombus formation in mast cell-deficient W/Wv mice.

We used mast cell-deficient W/Wv mice to clarify whether mast cell-derived heparin may play a role in inhibiting thrombus formation in the living organism. Small veins in the mesentery of W/Wv or congenic +/+ mice were stretched over an inverted microscope; a micropipette filled with varying concentrations of ADP was set close to the outside of a vein by using a micromanipulator. Thrombus formation was directly examined under the microscope. The concentration of ADP necessary for thrombus formation was significantly lower in the W/Wv mice than in the congenic +/+ mice. Furthermore, the concentration of ADP necessary for aggregation of platelets in platelet-rich plasma (PRP) was significantly lower in W/Wv mice than in +/+ mice. The higher sensitivity of PRP of W/Wv mice is not attributed to the platelets, but to the plasma, since platelets of +/+ mice suspended in platelet-poor plasma (PPP) of W/Wv mice were more sensitive to ADP than platelets of W/Wv mice suspended in PPP of +/+ mice. The present results suggest that plasma of W/Wv mice may lack any inhibitory factor(s) or contain promoting factor(s) for platelet aggregation.