Evaluation of Phosphorus Enrichment in Groundwater by Legacy Phosphorus in Orchard Soils with High Phosphorus Adsorption Capacity Using Phosphate Oxygen Isotope Analysis.

Long-term phosphorus (P) fertilization results in P accumulation in agricultural soil and increases the risk of P leaching into water bodies. However, evaluating P leaching into groundwater is challenging, especially in clay soil with a high P sorption capacity. This study examined whether the combination of PO4 oxygen isotope (δ18OPO4) analysis and the P saturation ratio (PSR) was useful to identify P enrichment mechanisms in groundwater. We investigated the groundwater and possible P sources in Kubi, western Japan, with intensive citrus cultivation. Shallow groundwater had oxic conditions with high PO4 concentrations, and orchard soil P accumulation was high compared with forest soil. Although the soil had a high P sorption capacity, the PSR was above the threshold, indicating a high risk of P leaching from the surface orchard soil. The shallow groundwater δ18OPO4 values were higher than the expected isotopic equilibrium with pyrophosphatase. The high PSR and δ18OPO4 orchard soil values indicated that P leaching from orchard soil was the major P enrichment mechanism. The Bayesian mixing model estimated that 76.6% of the P supplied from the orchard soil was recycled by microorganisms. This demonstrates the utility of δ18OPO4 and the PSR to evaluate the P source and biological recycling in groundwater.

Assessment of nitrogen budget in detailed spatial pattern using high precision modeling approach with constructed accurate agricultural behavior.

Changes in the nitrogen cycle due to fertilizer use can cause severe environmental pollution, particularly groundwater pollution, and threaten biosphere integrity. There are many difficulties and limitations in assessing groundwater pollution and a detailed nitrogen budget in an agricultural catchment. Previous methodologies have failed in an accurate assessment of the nitrogen budget in detailed spatial patterns. Herein, we designed a new modeling approach to assess the nitrogen budget using detailed spatial patterns in an agricultural catchment in the Nara Basin. We revised the Soil and Water Assessment Tool file output format, added the results for river nutrient concentrations and ammonia volatilization to the original output file. In this study, we calibrated and validated crop harvests, paddy evapotranspiration, streamflow, and river water concentrations of nitrate-nitrogen and total nitrogen to improve model accuracy as much as possible. Among them, data for evapotranspiration was obtained from a newly released Landsat dataset. The results showed that the amount of nitrogen leaching in rice paddies was 42 kg/ha, accounting for 65 % of total leaching in the study catchment. Cambisols and Fluvic Gleysols were prone to denitrification, and nitrogen leaching or denitrification occurred relatively more readily in low-slope areas. Furthermore, a detailed analysis of nitrogen cycle processes with high spatial precision indicates that areas with severe surface water pollution may also exhibit significant groundwater pollution. Our findings provide new solutions for assessing the nitrogen budget and groundwater pollution in catchments.

Spatial and temporal dynamics and fluxes estimation of manganese fractions in sediments from the Pearl River Estuary, southern China.

Sequential extraction was used to study the historical dynamics and fluxes of the chemical fractions of manganese (Mn) in sediments collected from the Pearl River Estuary (PRE), southern China. Results revealed that the proportion of Mn associated with different fractions decreased in the order of acid-soluble fraction (F1) > reducible fraction (F2) > residual fraction (F4) > oxidizable fraction (F3). F1 (47%) was the dominant Mn fraction, indicating the strong bioavailability of Mn to aquatic organisms in the PRE. In addition, the Mn fraction F2 was present at an average rate of 27.93 % in surface sediments, indicating that it could be mobilized when environmental conditions become increasingly reducing or oxidizing. The decline in Mn fraction fluxes after 2006 indicated that the region has partially decreased due to the removal of heavily polluting industries and effective control of sewage discharge.

Spatial and Temporal Variations and Risk Assessment of Heavy Metal Fractions in Sediments of the Pearl River Estuary, Southern China.

Sequential extraction was used to study the mobility and ecological risk of chemical fractions of six heavy metals in sediments collected from the Pearl River Delta (PRE) in China. Results revealed that residual fractions (F4) were the dominant forms for Cr and Ni in surface sediments, indicating that they were primarily stable in nature and had low bioavailability and ecotoxicity. Cd had a high environmental risk owing to its higher availability in acid-soluble fraction (F1), whereas Pb occurred predominantly in the reducible fraction (F2) in surface sediments. The profile variations of bioavailable fractions were generally consistent with socioeconomic development in the Pearl River Delta (PRD). A decreasing trend after 2006 suggested a reduction in heavy metal bioavailable fractions owing to the removal of heavy polluting industries and the effective control of sewage discharge. The risk assessment code suggested that the high mobility of Cd posed an extremely high risk and a threat to the aquatic environment.

A new sampling method with zirconium-loaded resin for phosphate oxygen isotope analysis in oligotrophic freshwater systems.

RATIONALE: The phosphate oxygen isotope ratio ( δ 18 O PO 4 ) is a useful technique to trace the sources and biogeochemical cycles of phosphorus (P) in aquatic ecosystems. However, δ 18 O PO 4 has not been widely used in oligotrophic freshwater systems due to technical and methodological difficulties in collecting sufficient phosphate (PO4 ) for the δ 18 O PO 4 analysis, which sometimes requires hundreds of liters of the water sample. In this study, a new approach (PaS-Zir) was developed for the δ 18 O PO 4 analysis in oligotrophic freshwater systems using zirconium (Zr)-loaded (ZrIRC) resin, which has a high affinity for PO4 . METHODS: ZrClO2 was added to Amberlite IRC748 to obtain the ZrIRC resin. The adsorption/desorption experiment using KH2 PO4 with a known value of δ 18 O PO 4 was conducted to determine the adsorption/desorption properties of the resin and the likelihood of isotopic fractionation. By installing mesh bags filled with the resin, the PaS-Zir approach was used in two rivers with low PO4 concentrations (0.2 and 5.3 µmol/L). A conventional sampling method was also performed in the study river with a higher PO4 concentration to validate the efficacy of the PaS-Zir method. RESULTS: The adsorption/desorption experiment demonstrated that the ZrIRC resin possessed a sufficient adsorption capacity (153 µmol/resin-mL) and exhibited little isotopic fractionation during the adsorption/desorption processes. Using the PaS-Zir method, we were able to collect sufficient PO4 samples for the δ 18 O PO 4 analysis from the rivers within at least 4 days of mesh bag installation. The δ 18 O PO 4 values (14.2‰ ± 0.2‰) obtained using the PaS-Zir method were comparable to those obtained using the conventional method (14.0‰ ± 0.03‰). CONCLUSION: We proved that the PaS-Zir method is applicable to oligotrophic freshwater systems and is generally more efficient than the conventional method. In addition, our method is useful for improving the understanding of the P dynamics of oligotrophic ecosystems because of the extremely low concentration of PO4 commonly found in them, which are often prone to P pollution.

A new method for phosphate purification for oxygen isotope ratio analysis in freshwater and soil extracts using solid-phase extraction with zirconium-loaded resin.

RATIONALE: Phosphate (PO4 ) oxygen isotope (δ18 OPO4 ) analysis is increasingly applied to elucidate phosphorus cycling. Due to its usefulness, analytical methods continue to be developed and improved to increase processing efficiency and applicability to various sample types. A new pretreatment procedure to obtain clean Ag3 PO4 using solid-phase extraction (SPE) with zirconium-loaded resin (ZrME), which can selectively adsorb PO4 , is presented and evaluated here. METHODS: Our method comprises (1) PO4 concentration, (2) PO4 separation by SPE, (3) cation removal, (4) Cl- removal, and (5) formation of Ag3 PO4 . The method was tested by comparing the resulting δ18 OPO4 of KH2 PO4 reagent, soil extracts (NaHCO3 , NaOH, and HCl), freshwater, and seawater with data obtained using a conventional pretreatment method. RESULTS: PO4 recovery of our method ranged from 79.2% to 97.8% for KH2 PO4 , soil extracts, and freshwater. Although the recovery rate indicated incomplete desorption of PO4 from the ZrME columns, our method produced high-purity Ag3 PO4 and accurate δ18 OPO4 values (i.e., consistent with those obtained using conventional pretreatment methods). However, for seawater, the PO4 recovery was low (1.1%), probably due to the high concentrations of F- and SO4 2- which interfere with PO4 adsorption on the columns. Experiments indicate that the ZrME columns could be regenerated and used repeatedly at least three times. CONCLUSIONS: We demonstrated the utility of ZrME for purification of PO4 from freshwater and soil extracts for δ18 OPO4 analysis. Multiple samples could be processed in three days using this method, increasing sample throughput and potentially facilitating more widespread use of δ18 OPO4 analysis to deepen our understanding of phosphorus cycling in natural environments.

Assessment of long-term phosphorus budget changes influenced by anthropogenic factors in a coastal catchment of Osaka Bay.

Phosphorus usage is irreplaceable in agriculture; however, its excessive use leads to wastage of invaluable resources and significant soil surplus. Agronomic soil phosphorus surplus in Asian regions has a much higher level than the global average. And with rapid urbanization and population growth in the recent decades, Asian countries have seen a rise in environmental pollution levels also. This study assessed the detailed phosphorus budget in the Yamato River catchment, an urbanized coastal catchment in Asia, from 1940s to 2010s using Soil and Water Assessment Tool, comprehensively analyzed the effect of anthropogenic factors on long-term phosphorus loading and agronomic soil phosphorus balance. The results showed the peak period of total phosphorus loading and agronomic soil phosphorus surplus occurred in 1970s, at 895 tons/year and 36.6 kg/ha, respectively. The major reasons for increased phosphorus loading and soil surplus during 1940-1970 were rapid population growth and increased fertilizer usage, respectively. Since the 1980s, the construction of wastewater treatment system and reduction in agricultural land contributed to environmental improvement. These anthropogenic factors had a much stronger impact on phosphorus budget than climate change in the study catchment. Soil phosphorus balance is affected by a combination of factors, such as soil properties, fertilizer usage and applied schedule, precipitation event, and crop types. And soil phosphorus surplus may be severely overestimated if the non-point source loss due to precipitation factor is not fully considered.

Effects of forest growth in different vegetation communities on forest catchment water balance.

Forest ecosystems are critical for adjusting the dynamic balance of the hydrological cycle. This balance is affected by vegetation community types, phenology, and forest density. Previous long-term catchment-scale model studies have focused on changes in forest areas while ignoring the above factors. Since the 1980s, climate change caused by increases in atmospheric CO2 levels has enhanced forest growth. Moreover, amendments to forest management policies, including intermediate cuttings caused by economic factors, have yielded unprecedented changes in forest ecosystems. In this study, we designed a methodology and created a credible model using the Soil and Water Assessment Tool (SWAT) that can precisely reflect water balance variations caused by different ecosystem situations during long-term changes in forest density. We focused on the Yamato River catchment in Western Japan, which includes three planted forests and one primeval forest, each markedly different with respect to vegetation community composition and management policy. In the process, we examined the ratio of coniferous vegetation and broad-leaved vegetation in different forest areas, used remote sensing methods to quantify the maximum and minimum leaf area index (LAI) of each forest region over 40 years, and calibrated the model by comparing the LAI growth curve, evapotranspiration, and streamflow with observed data. Moreover, we separated the decadal canopy evaporation, transpiration, and soil evaporation from the SWAT output results. We found that (1) forest evapotranspiration has increased in recent decades because of the above reasons; (2) in young or well-managed forests, the forest water balance may have changed significantly with forest growth. For long-term studies, it is necessary to distinguish the growth characteristics of different forests during different periods, and a detailed definition of a mixed forest is required. The forest parameters and growth characteristics are critical for understanding forest ecosystems and cannot be ignored at catchment-scale.

210Pb dating to investigate the historical variations and identification of different sources of heavy metal pollution in sediments of the Pearl River Estuary, Southern China.

In this study, we investigated the historical variation, source identification, and distribution of heavy metal pollution in sediments of the Pearl River Estuary (PRE) using 210Pb dating. Our results suggest that the heavy metal concentrations were higher in the western part of the estuary. For all heavy metals, Cd was significantly enriched in the sediments. The Pearl River Delta (PRD) has experienced rapid economic development in the past 40 years, a decreasing trend in heavy metal fluxes after 2004 was identified, which suggests a reduction in heavy metal concentrations due to the removal of heavy polluting industries and the effective control of sewage discharge. A binary mixing model reveals that the contributions of anthropogenic Pb ranged from 45.4 to 64%. Based on lead isotopic ratios (206/207Pb and 208/206Pb), it was found that geologic materials and industrial pollution were the main sources of heavy metals in the PRE sediments.

Hypoelectrolytic isoosmotic solution for infusion prevents saline-induced ultrastuructural artifacts of renal biopsy specimens.

Artifacts in the process of specimen preparation are frequent in ultrastructural evaluation of renal biopsy. We hypothesized that the common practice of wrapping kidney biopsy specimens in saline-soaked gauze to prevent the drying of the specimens could be the major factor of artifacts. In this study, whole kidneys from two male Sprague-Dawley rats were used. Before fixation, fresh small cubes of kidney tissue were macerated in saline (Saline group) or hypoelectrolytic isoosmotic solution for infusion (HISI group) (Sorita T3 or SOLDEM 3A) for 10 or 30 min. Then, the specimens were processed by 1% OsO(4) in 0.1 M phosphate buffer (pH 7.4) and embedded by EPON 812 for ultramicroscopic analysis. In the Saline group, ultrastructural examination revealed swollen podocyte, swollen capillary protuberance of the mesangium into the glomerular capillary loop, tubular cells with swollen mitochondria and microvilli, and the smooth muscle cells in the arteriolar wall with marked vacuolar degeneration were detected after 10 min maceration in saline and these findings become more pronounced after 30 min maceration. However, in the HISI group, these artifacts were not identified or limited within 30 min. It is postulated that HISI solution could prevent the artifacts, and be used for soaking and wrapping instead of physiologic saline solution.

The morphology and functions of articular chondrocytes on a honeycomb-patterned surface.

The present study investigated the potential of a novel micropatterned substrate for neocartilage formation. Articular chondrocytes were cultured on poly( ɛ-caprolactone) materials whose surfaces were either flat or honeycomb-patterned. The latter was prepared using a novel self-organization technique, while the former, was prepared by spin-coating. The chondrocytes attached and proliferated on both surfaces. On the honeycomb films, chondrocytes were found at the top surface and encased within the 10 µm pores. Meanwhile, chondrocytes on the spin-coated surface flattened out. Accumulation of DNA and keratin sulphate was comparatively higher on the honeycomb films within the first 7 days. At their respective peaks, DNA concentration increased on the honeycomb and flat surfaces by approximately 210% and 400% of their day 1 values, respectively. However, cultures on the flat surface took longer to peak. Extracellular Matrix (ECM) concentrations peaked at 900% and 320% increases for the honeycomb and flat cultures. Type II collagen was upregulated on the honeycomb and flat surfaces by as much as 28% and 25% of their day 1 values, while aggrecan was downregulated with time, by 3.4% and 7.4%. These initial results demonstrate the potential usefulness of honeycomb-based scaffolds during early cultures neocartilage and soft tissue engineering.

Gene expression profile of the cartilage tissue spontaneously regenerated in vivo by using a novel double-network gel: comparisons with the normal articular cartilage.

BACKGROUND: We have recently found a phenomenon that spontaneous regeneration of a hyaline cartilage-like tissue can be induced in a large osteochondral defect by implanting a double-network (DN) hydrogel plug, which was composed of poly-(2-Acrylamido-2-methylpropanesulfonic acid) and poly-(N, N'-Dimetyl acrylamide), at the bottom of the defect. The purpose of this study was to clarify gene expression profile of the regenerated tissue in comparison with that of the normal articular cartilage. METHODS: We created a cylindrical osteochondral defect in the rabbit femoral grooves. Then, we implanted the DN gel plug at the bottom of the defect. At 2 and 4 weeks after surgery, the regenerated tissue was analyzed using DNA microarray and immunohistochemical examinations. RESULTS: The gene expression profiles of the regenerated tissues were macroscopically similar to the normal cartilage, but showed some minor differences. The expression degree of COL2A1, COL1A2, COL10A1, DCN, FMOD, SPARC, FLOD2, CHAD, CTGF, and COMP genes was greater in the regenerated tissue than in the normal cartilage. The top 30 genes that expressed 5 times or more in the regenerated tissue as compared with the normal cartilage included type-2 collagen, type-10 collagen, FN, vimentin, COMP, EF1alpha, TFCP2, and GAPDH genes. CONCLUSIONS: The tissue regenerated by using the DN gel was genetically similar but not completely identical to articular cartilage. The genetic data shown in this study are useful for future studies to identify specific genes involved in spontaneous cartilage regeneration.

Induction of spontaneous hyaline cartilage regeneration using a double-network gel: efficacy of a novel therapeutic strategy for an articular cartilage defect.

BACKGROUND: A double-network (DN) gel, which was composed of poly-(2-acrylamido-2-methylpropanesulfonic acid) and poly-(N,N'-dimetyl acrylamide) (PAMPS/PDMAAm), has the potential to induce chondrogenesis both in vitro and in vivo. PURPOSE: To establish the efficacy of a therapeutic strategy for an articular cartilage defect using a DN gel. STUDY DESIGN: Controlled laboratory study. METHODS: A 4.3-mm-diameter osteochondral defect was created in rabbit trochlea. A DN gel plug was implanted into the defect of the right knee so that a defect 2 mm in depth remained after surgery. An untreated defect of the left knee provided control data. The osteochondral defects created were examined by histological and immunohistochemical evaluations, surface assessment using confocal laser scanning microscopy, and real-time polymerase chain reaction (PCR) analysis at 4 and 12 weeks. Samples were quantitatively evaluated with 2 scoring systems reported by Wayne et al and O'Driscoll et al. RESULTS: The DN gel-implanted defect was filled with a sufficient volume of the hyaline cartilage tissue rich in proteoglycan and type 2 collagen. Quantitative evaluation using the grading scales revealed a significantly higher score in the DN gel-implanted defects compared with the untreated control at each period (P < .0001). The mean relative values of type 2 collagen mRNAs in the regenerated tissue were obviously higher in the DN gel-implanted defect than in the untreated control at each period. The mean surface roughness of the untreated control was significantly higher than the normal cartilage at 12 weeks (P = .0106), while there was no statistical difference between the DN gel-implanted and normal knees. CONCLUSION: This study using the mature rabbit femoral trochlea osteochondral defect model demonstrated that DN gel implantation is an effective treatment to induce cartilage regeneration in vivo without any cultured cells or mammalian-derived scaffolds. CLINICAL RELEVANCE: This study has prompted us to develop a potential innovative strategy to repair cartilage lesions in the field of joint surgery.

Spontaneous hyaline cartilage regeneration can be induced in an osteochondral defect created in the femoral condyle using a novel double-network hydrogel.

BACKGROUND: Functional repair of articular osteochondral defects remains a major challenge not only in the field of knee surgery but also in tissue regeneration medicine. The purpose is to clarify whether the spontaneous hyaline cartilage regeneration can be induced in a large osteochondral defect created in the femoral condyle by means of implanting a novel double-network (DN) gel at the bottom of the defect. METHODS: Twenty-five mature rabbits were used in this study. In the bilateral knees of each animal, we created an osteochondral defect having a diameter of 2.4-mm in the medial condyle. Then, in 21 rabbits, we implanted a DN gel plug into a right knee defect so that a vacant space of 1.5-mm depth (in Group I), 2.5-mm depth (in Group II), or 3.5-mm depth (in Group III) was left. In the left knee, we did not apply any treatment to the defect to obtain the control data. All the rabbits were sacrificed at 4 weeks, and the gross and histological evaluations were performed. The remaining 4 rabbits underwent the same treatment as used in Group II, and real-time PCR analysis was performed at 4 weeks. RESULTS: The defect in Group II was filled with a sufficient volume of the hyaline cartilage tissue rich in proteoglycan and type-2 collagen. The Wayne's gross appearance and histology scores showed that Group II was significantly greater than Group I, III, and Control (p < 0.012). The relative expression level of type-2 collagen, aggrecan, and SOX9 mRNAs was significantly greater in Group II than in the control group (p < 0.023). CONCLUSIONS: This study demonstrated that spontaneous hyaline cartilage regeneration can be induced in vivo in an osteochondral defect created in the femoral condyle by means of implanting the DN gel plug at the bottom of the defect so that an approximately 2-mm deep vacant space was intentionally left in the defect. This fact has prompted us to propose an innovative strategy without cell culture to repair osteochondral lesions in the femoral condyle.

Joint immobilization inhibits spontaneous hyaline cartilage regeneration induced by a novel double-network gel implantation.

We have recently discovered that spontaneous hyaline cartilage regeneration can be induced in an osteochondral defect in the rabbit, when we implant a novel double-network (DN) gel plug at the bottom of the defect. To clarify whether joint immobilization inhibits the spontaneous hyaline cartilage regeneration, we conducted this study with 20 rabbits. At 4 or 12 weeks after surgery, the defect in the mobile knees was filled with a sufficient volume of the hyaline cartilage tissue rich in proteoglycan and type-2 collagen, while no cartilage tissues were observed in the defect in the immobilized knees. Type-2 collagen, Aggrecan, and SOX9 mRNAs were expressed only in the mobile knees at each period. This study demonstrated that joint immobilization significantly inhibits the spontaneous hyaline cartilage regeneration induced by the DN gel implantation. This fact suggested that the mechanical environment is one of the significant factors to induce this phenomenon.

In vivo imaging of particle-induced inflammation and osteolysis in the calvariae of NFκB/luciferase transgenic mice.

Wear debris causes biological response which can result in periprosthetic osteolysis after total joint replacement surgery. Nuclear factor-kappa B (NFκB), a representative transcription factor involved in inflammation, is believed to play an important role in this event by regulating the production of proinflammatory mediators and osteoclastogenesis. In this study, we sought to determine whether activation of NFκB in response to stimulation by particles could be visualized by in vivo imaging. We loaded polyethylene (PE) particles onto the calvaria of NFκB/luciferase transgenic mouse, and detected luminescence generated by activation of NFκB. On day 7 after loading, the level of luminescence was maximal. Levels of luminescence were significantly correlated with the levels of luciferase activity, proinflammatory mediator mRNAs, and bone resorption parameters. This system, which enabled us to evaluate particle-induced inflammation and osteolysis without sacrificing mice, constitutes a useful tool for evaluating the efficacy of prophylaxis or treatments for particle-induced osteolysis.

Deficiency of macrophage migration inhibitory factor gene delays healing of the medial collateral ligament: a biomechanical and biological study.

The role played by macrophage migration inhibitory factor (MIF) in the process of wound healing is controversial. Besides, there have been no reports that investigated the expression or the role of MIF in the repair process after ligament injury. In this study, we hypothesized that the deficiency in MIF gene might delay ligament healing in mice. The aim of this study was to clarify this hypothesis using MIF gene-deficient mice (MIFKO) and murine model of injury to the medial collateral ligament (MCL). Biomechanical testing showed that the levels of mechanical properties were significantly lower in MIFKO than in wild-type mice (WT) on day 28 after injury. Levels of matrix metalloproteinase (MMP)-2 and -13 mRNA in the healing tissue were significantly lower in MIFKO than in WT on day 28 and on day 7, respectively. Histologically, healing tissues in MIFKO exhibited prolonged hypertrophy, poor vascularity, and prolonged increase in cell number compared with those in WT. Taken together, it was suggested that MIFKO exhibited delayed healing of the MCL, which might be caused by lower mRNA expression of MMP-2 and -13.

Gene expression, glycocalyx assay, and surface properties of human endothelial cells cultured on hydrogel matrix with sulfonic moiety: Effect of elasticity of hydrogel.

We measured the gene expression, glycocalyx content, and surface properties of human coronary artery endothelial cells (HCAECs) cultured on poly(sodium p-styrene sulfonate) (PNaSS) hydrogels with various levels of elasticity ranged in 3-300 kPa. We found that all HCAECs reached confluence on these hydrogels while retaining the similar expression of EC-specific markers to that on polystyrene (PS), a widely used scaffold in cell culture in vitro. Real-time polymerase chain reaction (PCR) and glycosaminoglycan (GAG) assay showed that the amount of EC-specific glycocalyx secreted by HCAECs cultured on PNaSS gels was higher than that cultured on PS, and it increased with an increase of gel elasticity. Furthermore, the HCAECs cultured on PNaSS gels showed excellent property against platelet adhesion and lower surface friction than that on PS. The platelet adhesion and surface friction of HCAECs cultured on PNaSS gels also depend on the elasticity of gels. The largest amount of EC-specific glycocalyx, excellent blood compatibility, and the lowest friction were observed when the elastic modulus of the gel was larger than 60 kPa. Overall, HCAECs cultured on these hydrogels have better properties than those cultured on PS scaffold, demonstrating the PNaSS gels can be used as potential tissue engineering material for blood vessels.

Effects of environmental regulations on heavy metal pollution decline in core sediments from Manila Bay.

We investigated the high-resolution heavy metal pollution history of Manila Bay using heavy metal concentrations and Pb isotope ratios together with (210)Pb dating to find out the effects of environmental regulations after the 1990 s. Our results suggested that the rate of decline in heavy metal pollution increased dramatically from the end of the 1990 s due to stricter environmental regulations, Administrative Order No. 42, being enforced by the Philippines government. The presented data and methodology should form the basis for future monitoring, leading to pollution control, and to the generation of preventive measures at the pollution source for the maintenance of environmental quality in the coastal metropolitan city of Manila. Although this is the first report of a reduction in pollution in Asian developing country, our results suggest that we can expect to find similar signs of pollution decline in other parts of the world as well.

DNA vaccination against macrophage migration inhibitory factor improves atopic dermatitis in murine models.

BACKGROUND: Atopic dermatitis (AD) is a common chronic inflammatory skin disease. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that has been implicated in the pathogenesis of AD. Recently, we developed a novel DNA vaccine that generates neutralizing endogenous anti-MIF antibodies. OBJECTIVE: This study explores the preventive and therapeutic effects of this MIF-DNA vaccine in mouse models of AD. METHODS: Two different AD model mice (DS-Nh and NC/Nga) received MIF-DNA vaccination to analyze preventive and therapeutic effects, as assessed by clinical skin scores, histologic findings, and serum IgE levels. RESULTS: In murine models of AD, MIF-DNA vaccination prevented the occurrence of the AD skin phenotype. Furthermore, administration of MIF-DNA vaccine to mice that had already developed AD produced a rapid improvement in AD skin manifestation. There were reduced histologic signs of inflammation and lower serum IgE levels in treated mice compared with those seen in control animals. Finally, passive transfer of IgG from MIF-DNA vaccinated mice to AD mice also produced a significant therapeutic effect. These results demonstrate that MIF-DNA vaccination not only prevents the development of AD but also improves the symptoms of pre-existing AD. CONCLUSION: Taken together, the induction of an anti-MIF autoantibody response using MIF-DNA vaccination appears to be a useful approach in the treatment of AD.