Development of an Analytical Method for Nitric Oxide Radical Determination in Natural Waters

The measurement of photochemically generated nitric oxide radicals (NO) in natural waters has long been an arduous task because of a lack of simple analytical techniques, even though the environmental significance of this radical is paramount. We have developed a simple analytical method for the determination of photochemically generated NO in natural waters using 4,5-diaminofluorescein (DAF-2) as a probe compound. This method is based on the reaction of photoformed NO with DAF-2 in air-saturated solution to produce a highly fluorescent triazolofluorescein (DAF-2T) product. DAF-2T was determined by using reversed-phase HPLC with fluorescence detection, with excitation and emission wavelengths of 495 and 515 nm, respectively. Under optimum conditions, the calibration curve exhibited linearity in the range of 0.025−10 nM DAF-2T. The coefficients of variance for the measurements of the signal intensities of DAF-2T (from the photolysis of 0.5 μM and 5 μM NO2− with DAF-2) were less than 5% and 3%, respectively. For a total irradiation time of 30 min, the detection limit of the photoformation rate of NO was 1.65 × 10−13 M s−1, defined as 3σ of the lowest measured DAF-2T concentration (0.025 nM). The proposed method is relatively unaffected by potential interferents in seawater. The method was employed to determine the photoformation rate of NO in the Seto Inland Sea and the Kurose River in Hiroshima Prefecture, Japan. The measured NO photoformation rates in seawater and river water samples ranged from (5.3−32) × 10−12 M s−1 and (9.4−300) × 10−12 M s−1, respectively

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

Application of Fenton reaction for nanomolar determination of hydrogen peroxide in seawater

A simple and sensitive method for the determination of nanomolar levels of hydrogen peroxide (H2O2) in seawater has been developed and validated. This method is based on the reduction of H2O2 by ferrous iron in acid solution to yield hydroxyl radical (•OH) which reacts with benzene to produce phenol. Phenol is separated from the reaction mixture by reversed phase high performance liquid chromatography and its fluorescence intensity signals were measured at excitation and emission of 270 and 298 nm, respectively. Under optimum conditions, the calibration curve exhibited linearity in the range of (0–50) × 103 nmol L−1 H2O2. The relative standard deviations for five replicate measurements of 500 and 50 nmol L−1 H2O2 are 1.9 and 2.4%, respectively. The detection limit for H2O2, defined as three times the standard deviation of the lowest standard solution (5 nmol L−1 H2O2) in seawater is 4 nmol L−1. Interference of nitrite ion (NO2−) on the fluorescence intensity of phenol was also investigated. The result indicated that the addition of 10mol L−1 NO2− to seawater samples showed no significant interference, although, the addition of 50mol L−1 NO2 − to the seawater samples decreases the fluorescence intensity signals of phenol by almost 40%. Intercomparison of this method with well-accepted (p-hydroxyphenyl) acetic acid (POHPAA)-FIA method shows excellent agreement. The proposed method has been applied on-board analysis of H2O2 in Seto Inland seawater samples

Periphyton contribution to nitrogen dynamics in the discharge from a wastewater treatment plant

To evaluate the importance of periphyton to nitrogen dynamics in the discharge from wastewater treatment plants (WWTPs), we examined changes in total and inorganic nitrogen content downstream from a WWTP on the Kurose River in Hiroshima Prefecture, Japan. At 0.7 km downstream of the WWTP (point A), NH4+-N was the dominant form of inorganic nitrogen, but concentrations decreased rapidly to 5 km downstream (point B). In contrast, no significant change in the [NO2– + NO3–]-N concentration was observed between the two points. Total nitrogen (TN) load decreased significantly between the two points, suggesting that sorption and/or denitrification occurred in the river channel. Potential rates of nitrogen sorption and transformation by periphyton were determined in a laboratory experiment in which changes in the nitrogen content of river water were examined in an acrylic chamber with periphyton. Nitrification and nitrogen removal occurred mainly in the periphyton. The contributions of periphyton activity to TN and NH4+-N decrease in the field, as estimated from the results of the laboratory experiments, were 6%–18% and 23%–72%, respectively. These results suggest that periphyton plays an important role in decreasing NH4+-N concentration in the discharge from wastewater treatment plants

A case of left ventricular free wall rupture after insertion of an IMPELLA® left ventricular assist device diagnosed by transesophageal echocardiography

[Background]The IMPELLA® is a minimally invasive left ventricular assist device. We report a case in which transesophageal echocardiography (TEE) was useful in diagnosis of left ventricular rupture after IMPELLA® insertion. [Case presentation]A 75-year-old man presented to the emergency room with chest pain and underwent percutaneous coronary intervention for 100% stenosis of the left anterior descending branch #7. An IMPELLA® was inserted to stabilize the circulation, but hypotension persisted. Transthoracic echocardiography revealed increased pericardial effusion and suspicion of free wall left ventricular rupture, leading to emergency surgery. TEE revealed the IMPELLA® straying into the left ventricle apical wall and cardiac tamponade. Hemorrhage was observed from the thinning free wall and the tip of the IMPELLA® was palpable. The IMPELLA® was removed and the left ventricular wall was repaired. [Conclusions]The IMPELLA® requires implantation of the tip in the left ventricle, but it should be noted that a fragile ventricular wall can be easily perforated

Mutational study of sapovirus expression in insect cells

Human sapovirus (SaV), an agent of human gastroenteritis, cannot be grown in cell culture, but expression of the recombinant capsid protein (rVP1) in a baculovirus expression system results in the formation of virus-like particles (VLPs). In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence (Wt construct), whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent (MEG-1076 construct). While both constructs formed VLPs morphologically similar to native SaV, Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore, Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly, the position of the mutated residue was strictly conserved residue among other human SaV strains, suggesting an important role for rVP1 expression