The Circadian Clock in the Retinal Pigment Epithelium Controls the Diurnal Rhythm of Phagocytic Activity.

The diurnal peak of phagocytosis by the retinal pigment epithelium (RPE) of photoreceptor outer segments (POS) is under circadian control and believed that this process involves interactions from the retina and RPE. Previous studies have demonstrated that a functional circadian clock exists within multiple retinal cell types and RPE. Thereby, the aim of this study was to determine whether the clock in the retina or RPE controls the diurnal phagocytic peak and whether disruption of the circadian clock in the RPE would affect cellular function and the viability during aging. To that, we generated and validated an RPE tissue-specific KO of the essential clock gene, Bmal1, and then determined the daily rhythm in phagocytic activity by the RPE in mice lacking a functional circadian clock in the retina or RPE. Then, using electroretinography, spectral domain-optical coherence tomography, and optomotor response of visual function we determined the effect of Bmal1 removal in young (6 months) and old (18 months) mice. RPE morphology and lipofuscin accumulation was determined in young and old mice. Our data shows that the clock in the RPE, rather than the retina clock, controls the diurnal phagocytic peak. Surprisingly, absence of a functional RPE clock and phagocytic peak does not result in any detectable age-related degenerative phenotype in the retina or RPE. Thus, our results demonstrate that the circadian clock in the RPE controls the daily peak of phagocytic activity. However, the absence of the clock in the RPE does not result in deterioration of photoreceptors or the RPE during aging.

[Clinical significance of oxidized low-density lipoprotein antibody in antiphospholipid syndrome].

OBJECTIVE: To investigate the significance and distribution of oxidized low-density lipoprotein antibodies (ox-LDL-Ab) in patients with antiphospholipid syndrome (APS). METHODS: In this study, 334 patients who were hospitalized in the Department of Rheumatology and Immunology, Peking University People's Hospital were included. There were 162 APS patients, 122 patients with other autoimmune diseases without thrombosis or obstetric disease as disease control and 50 healthy controls. The clinical data and laboratory indicators were retrospectively collected. The ox-LDL-Ab, anticardiolipin (aCL) IgG/IgA/IgM, and anti-ß2-glycoprotein Ⅰ (aß2GPI) IgG/IgA/IgM were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between ox-LDL-Ab and clinical and laboratory parameters were analyzed by SPSS 27.0. RESULTS: In APS group, 60.5% of patients had thrombosis, 48.1% had pregnancy morbidity, 34.0% had thrombocytopenia. The positive rates of aCL, aß2GPI and lupus anticoagulant (LAC) were 17.9%, 34.6%, and 46.9%, respectively. The ox-LDL-Ab titers and positive rate in APS group were higher than that in healthy controls [titers: 40.8 (25.4-66.0) U/mL vs. 24.1 (12.3-36.5) U/mL, P=0.001; positive rate: 67.3% vs. 36.0%, P=0.001]. The diffe-rences in titers and positive rate of ox-LDL-Ab between APS patients and disease controls were not statistically significant [titers: 40.8 (25.4-66.0) U/mL vs. 35.9 (24.2-53.1) U/mL, P=0.118; positive rate: 67.3% vs. 61.5%, P=0.318]. The area under curve (AUC) for aß2GPI, aCL, and ox-LDL-Ab were 0.745 (95%CI: 0.692-0.797), 0.666 (95%CI: 0.608-0.724), 0.609 (95%CI: 0.549-0.669), respectively. The Youden's index was 0.388, 0.269, and 0.132, respectively. The AUC for ox-LDL-Ab in seronegative APS patients was 0.562 (95%CI: 0.480-0.645). The sensitivity and specificity of ox-LDL-Ab in seronegative APS patients were 63.9% and 47.0%, respectively, and the Youden's index was 0.109. The ox-LDL-Ab positive group had higher positive rate of aß2GPI (42.2% vs. 18.9%, P=0.003) and aCL (22.9% vs. 7.5%, P=0.017) than the ox-LDL-Ab negative group. There was no correlation between ox-LDL-Ab and thrombosis, coronary artery disease, pregnancy morbidity, hyperlipidemia, hypocomplementemia, and LAC positivity. CONCLUSION: Ox-LDL-Ab was correlated with aCL and aß2GPI, and no association were observed between ox-LDL-Ab and thrombosis, coronary artery disease, and pregnancy morbidity.

Preoperative mean platelet volume and platelet distribution associated with thyroid cancer.

Thyroid cancer is the most frequent cancer of the endocrine glands and the fifth most frequent cancer in women. Activated platelets play a crucial role in thrombosis, inflammation, and cancer. Mean platelet volume (MPV) and platelet distribution width (PDW) are early index of platelet activation. The purpose of this study is to investigate platelet indices levels in thyroid cancer. The study enrolled 280 patients with thyroid cancer and 280 control subjects. Patients' characteristics and hematologic tests data were collected at the time of diagnosis. Correlations between platelet indices and clinical characteristics were analyzed. The odds ratios (ORs) and 95% confidence intervals (CIs) for thyroid cancer were calculated using multivariate logistic regression analyses across MPV and PDW quartiles. The patients with thyroid cancer had lower MPV and higher PDW compared with control subjects. MPV was correlated with tumor-nodus-metastases (TNM) stage and lymph node metastasis. Moreover, after adjusting for other risk factors, the prevalence risk of thyroid cancer for the lowest quartile of MPV was 7.242 (4.069-12.887) (P < 0.001) and for the highest quartile of PDW was 6.065 (3.321-11.076) (P < 0.001), respectively. The study showed that the patients with thyroid cancer have lower MPV and higher PDW compared to control subjects. Moreover, MPV and PDW were independently associated with the presence of thyroid cancer. Further studies are needed to evaluate the utility of MPV and PDW as novel diagnostic screening tools for thyroid cancer.

Conditional knockdown of DNA methyltransferase 1 reveals a key role of retinal pigment epithelium integrity in photoreceptor outer segment morphogenesis.

Dysfunction or death of photoreceptors is the primary cause of vision loss in retinal and macular degenerative diseases. As photoreceptors have an intimate relationship with the retinal pigment epithelium (RPE) for exchange of macromolecules, removal of shed membrane discs and retinoid recycling, an improved understanding of the development of the photoreceptor-RPE complex will allow better design of gene- and cell-based therapies. To explore the epigenetic contribution to retinal development we generated conditional knockout alleles of DNA methyltransferase 1 (Dnmt1) in mice. Conditional Dnmt1 knockdown in early eye development mediated by Rx-Cre did not produce lamination or cell fate defects, except in cones; however, the photoreceptors completely lacked outer segments despite near normal expression of phototransduction and cilia genes. We also identified disruption of RPE morphology and polarization as early as E15.5. Defects in outer segment biogenesis were evident with Dnmt1 exon excision only in RPE, but not when excision was directed exclusively to photoreceptors. We detected a reduction in DNA methylation of LINE1 elements (a measure of global DNA methylation) in developing mutant RPE as compared with neural retina, and of Tuba3a, which exhibited dramatically increased expression in mutant retina. These results demonstrate a unique function of DNMT1-mediated DNA methylation in controlling RPE apicobasal polarity and neural retina differentiation. We also establish a model to study the epigenetic mechanisms and signaling pathways that guide the modulation of photoreceptor outer segment morphogenesis by RPE during retinal development and disease.

Conditional Induction of Oxidative Stress in RPE: A Mouse Model of Progressive Retinal Degeneration.

An appropriate animal model is essential to screening drugs or designing a treatment strategy for geographic atrophy. Since oxidative stress contributes to the pathological changes of the retinal pigment epithelium (RPE), we are reporting a new mouse AMD model of retinal degeneration by inducing mitochondrial oxidative stress in RPE. Sod2 the gene for manganese superoxide dismutase (MnSOD) was deleted in RPE layer using conditional knockout strategy. Fundus microscopy, SD-OCT and electroretinography were used to monitor retinal structure and function in living animals and microscopy was used to assess pathology post mortem. Tissue specific deletion of Sod2 caused elevated signs of oxidative stress, RPE dysfunction and showed some key features of AMD. Due to induction of oxidative stress, the conditional knockout mice show progressive reduction in ERG responses and thinning of outer nuclear layer (ONL) compared to non-induced littermates.

Loss of mTOR signaling affects cone function, cone structure and expression of cone specific proteins without affecting cone survival.

Cones are the primary photoreceptor (PR) cells responsible for vision in humans. They are metabolically highly active requiring phosphoinositide 3-kinase (PI3K) activity for long-term survival. One of the downstream targets of PI3K is the kinase mammalian target of rapamycin (mTOR), which is a key regulator of cell metabolism and growth, integrating nutrient availability and growth factor signals. Both PI3K and mTOR are part of the insulin/mTOR signaling pathway, however if mTOR is required for long-term PR survival remains unknown. This is of particular interest since deregulation of this pathway in diabetes results in reduced PR function before the onset of any clinical signs of diabetic retinopathy. mTOR is found in two distinct complexes (mTORC1 & mTORC2) that are characterized by their unique accessory proteins RAPTOR and RICTOR respectively. mTORC1 regulates mainly cell metabolism in response to nutrient availability and growth factor signals, while mTORC2 regulates pro-survival mechanisms in response to growth factors. Here we analyze the effect on cones of loss of mTORC1, mTORC2 and simultaneous loss of mTORC1 & mTORC2. Interestingly, neither loss of mTORC1 nor mTORC2 affects cone function or survival at one year of age. However, outer and inner segment morphology is affected upon loss of either complex. In contrast, concurrent loss of mTORC1 and mTORC2 leads to a reduction in cone function without affecting cone viability. The data indicates that PI3K mediated pro-survival signals diverge upstream of both mTOR complexes in cones, suggesting that they are independent of mTOR activity. Furthermore, the data may help explain why PR function is reduced in diabetes, which can lead to deregulation of both mTOR complexes simultaneously. Finally, although mTOR is a key regulator of cell metabolism, and PRs are metabolically highly active, the data suggests that the role of mTOR in regulating the metabolic transcriptome in healthy cones is minimal.

Effect of p53 gene polymorphism on functions of prostate cancer cells.

Prostate cancer cells were transfected with plasmids [empty plasmids, wild-type pcDNA3.1-p53 (V/V), mutant type pcDNA3.1- p53 (G/G)] to analyze the effect of p53 gene polymorphisms on the proliferation, cycle, and apoptosis of prostatic cancer cells. Empty plasmids containing wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1- p53 (G/G) were used to transfect PC3 and LNCaP cells, respectively. Cell proliferation was detected at 0, 24, 48, and 72 h using the MTT method. Cells were collected at 24 and 72 h. The distribution of cell cycles in various groups was detected using flow cytometry (propidium iodide staining method) and the apoptosis rate was detected using annexin V + propidium iodide double staining. Compared with the control group, wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1-p53 (G/G) showed a significant inhibitory effect on cell proliferation (P < 0.05); the inhibitory effect of the mutant type was stronger than that of the wild-type. There was no significant difference between PC3 cells and LNCaP cells. After transfection with wild-type pcDNA3.1-p53 (V/V) and mutant type pcDNA3.1-p53 (G/G), PC3 and LNCaP cells were arrested in the G0/G1 stage. Transfection with pcDNA3.1-p53 (G/G) showed a more significant effect than transfection with pcDNA3.1-p53 (V/V). Both the wild-type pcDNA3.1-p53 (V/V) and mutant-type pcDNA3.1-p53 (G/G) led to an increased apoptosis rate of PC3 and LNCaP cells. The p53 gene polymorphism affects the proliferation, apoptosis, and cycle of prostate cancer cells and may serve as a reliable index for the diagnosis and treatment of prostate cancer.

The variation of volatile fatty acid compositions in sewer length, and its effect on the process design of biological nutrient removal.

The potential of enhanced biological phosphorus removal (EBPR) in biological nutrient removal (BNR) systems critically depends on the availability and types of volatile fatty acids (VFAs) in sewage. Although the characteristics of VFAs in sewage are strongly related with the biochemical transformations in the sewer system, they have not been studied thoroughly in terms of BNR process design. We have investigated the characteristics of VFAs in influent of nine sewage treatment plants which represent typical small to very large sewer systems in Korea. We found that influent total VFACOD (VFA as chemical oxygen demand) concentrations ranged from 20.4 to 65.2 mg/L. Acetic acid was a predominant VFA in sewage, and the propionic acid (HPr) portion averaged 38.7% of total VFACOD. However the sewage from longer sewer systems showed more HPr content, indicating that type of VFA varied with the total sewer length. The finding is a particularly important consideration for BNR process design since availability of HPr positively behaved to suppress the unfavorable growth of glycogen-accumulating organisms. The implication of these findings for BNR process design is discussed in this paper.

First-time donors responding to a national disaster may be an untapped resource for the blood centre.

BACKGROUND: On 12 May 2008, a severe earthquake struck Sichuan in China. Many people donated blood for the first time, leading us to question whether these donors might become repeat donors in the future. The return pattern of post-earthquake first-time donors (PEFTD) was compared with that of first-time donors (FTD) in a comparable period. METHODS: Demographic characteristics, transfusion-transmissible infection rates and 1-year return rates were compared between 5147 PEFTD (5/13-5/19, 2008) and 3176 FTD (5/13-5/19, 2009) from five Chinese blood centres using chi-squared tests. Adjusted logistic regression was used to detect earthquake effect on donor return. RESULTS: Post-earthquake first-time donors were more frequently between 26 and 45 years, men, and better educated compared with the control group. Slightly higher but not statistically significant increased rates of hepatitis B virus surface antigen (HBsAg) (0·87% vs. 0·50%, P=0·054), hepatitis C virus (HCV) (0·70% vs. 0·63%, P=0·414), syphilis (0·9% vs. 0·7%, P=0·489) and lower rates of human immunodeficiency virus (HIV) (0·31% vs. 0·60%, P=0·078) reactivity were detected for PEFTD. The 1-year return rate for PEFTD was significantly lower than that of the controls (8·0% vs. 13·0%, P<0·001). After adjusting for demographic factors, donation volume and sites, the PEFTD were less likely to return in 1 year than the controls (OR: 0·520; 95% CI: 0·442, 0·611). CONCLUSION: Post-earthquake first-time donors may be less likely to donate again without continuing motivation strategies. Further studies on PEFTD's lack of motivation to return for donation are needed to design recruitment strategies to convert PEFTD to become repeat donors to continuously replenish the blood supply.

Trafficking of membrane proteins to cone but not rod outer segments is dependent on heterotrimeric kinesin-II.

Heterotrimeric kinesin-II is a molecular motor localized to the inner segment, connecting cilium and axoneme of mammalian photoreceptors. Our purpose was to identify the role of kinesin-II in anterograde intraflagellar transport by photoreceptor-specific deletions of kinesin family member 3A (KIF3A), its obligatory motor subunit. In cones lacking KIF3A, membrane proteins involved in phototransduction did not traffic to the outer segments resulting in complete absence of a photopic electroretinogram and progressive cone degeneration. Rod photoreceptors lacking KIF3A degenerated rapidly between 2 and 4 weeks postnatally, but the phototransduction components including rhodopsin trafficked to the outer segments during the course of degeneration. Furthermore, KIF3A deletion did not affect synaptic anterograde trafficking. The results indicate that trafficking of membrane proteins to the outer segment is dependent on kinesin-II in cone, but not rod photoreceptors, even though rods and cones share similar structures, and closely related phototransduction polypeptides.

Psychological and behavioural impacts of the 2008 China earthquake on blood donors.

BACKGROUND AND OBJECTIVES: On May 12, 2008, a severe earthquake hit Sichuan province in China. A post-earthquake survey was conducted to study the earthquake's effect on blood donor behaviour and stress at three blood centres at varying distances from the epicentre. MATERIALS AND METHODS: A questionnaire was developed to assess donor post-traumatic stress disorders (PTSD) and attitudes toward giving blood. Responses were compared by centre and donor characteristics using multivariate logistic regression techniques. RESULTS: Of all 17 456 donors, the overall prevalence of PTSD was 13.2%. Donors who knew someone killed or injured by the earthquake were 2.1 times more likely to have PTSD than others (95% CI: 1.8-2.4). 85.2% of donors cited the earthquake as their reason for donating. 16.1% of donors felt it acceptable to be less honest about one's health history in an emergency. After adjusting for PTSD, geographic and demographic characteristics, the donors knowing someone killed or injured by the earthquake were 1.4 times (95% CI: 1.2-1.7) more likely to cite the earthquake as reason for donating, and 1.8 times (95% CI: 1.5-2.1) more likely to accept being less honest about one's health history in case of national emergency. CONCLUSIONS: The psychological and behavioural impacts of the earthquake on blood donors extended far from the epicentre. After a disaster, it is important to emphasize that donors must be truthful on the donor questionnaire as some donors appear willing to be less than honest when they perceive an increased need for blood products.

Poor outcome of adenovirus infections in adult hematopoietic stem cell transplant patients with sustained adenovirus viremia.

The outcome of adenovirus (ADV) infections in adult hematopoietic stem cell transplant (HSCT) patients remains poorly characterized. We studied 14 adults and 3 children, who had undergone HSCT and had developed ADV viremia. Peak ADV DNA levels were significantly higher in patients with ADV diseases than in those without (P=0.03). All children survived the ADV infections. Among the 14 adult HSCT patients, 11 were treated with cidofovir, 2 with ribavirin, and 1 did not receive antiviral treatment. Six of the 13 (46%) treated patients developed ADV diseases and 3 of them (23%) died of ADV infections. Sustained viremia (≥3 positive polymerase chain reaction assays during follow-up) was detected in all patients who finally died of ADV infections. However, 2 adults having had transient ADV viremia either survived or died of diseases other than ADV infections. Our study indicates that the outcome of adult HSCT patients with sustained ADV viremia may be poor, even for those who have received anti-ADV treatment.

Mathematical models for biomass in membrane-BNR process for wastewater treatment.

The equation of biomass is related to the mass-balance equation of substrate. This equation of substrate is expressed according to a model using the Monod equation, which indicates some limits for calculating the amounts of VSS in the MBR process. Some degradation of biomass which is caused by long SRT might result in the generation of substrate based on COD. Research was conducted by lab-scale tests with two membrane-BNR (Biological Nutrients Removal) processes. These were composed of multi-reactors as anaerobic, anoxic, aerobic tank and oxygen exhauster. The aerobic tank was also divided into 3 reactors, which were oxic for nitrification, oxic-media containing fluidized sponge typed media for simultaneous nitrification and denitrification, and oxic-membrane for submerged membrane. This membrane-BNR process could remove most of the organics, suspended solids and nutrient substances like nitrogen thus satisfying the reuse guidelines issued by the Korean Ministry of Environment. The value measured of VSS (X(v)) through the experiment with SRT of 35 days was similar to the biomass using the conventional equation while the one with SRT of 60 days was close to the concentration of VSS calculated by a revised equation which considered the biomass degraded with long SRT.

Effects of Cone Connexin-36 Disruption on Light Adaptation and Circadian Regulation of the Photopic ERG.

Purpose: The present study tested the hypothesis that connexin-36 (Cx36) and gap junctions between photoreceptor cells contribute to the circadian rhythm of the photopic electroretinogram (ERG) b-wave amplitude. Methods: Cone-specific disruption of Cx36 was obtained in mice with a floxed Gjd2 gene and human red/green pigment promoter (HRGP)-driven Cre recombinase. Standard ERG, spectral-domain optical coherence tomography (SD-OCT) and histochemical methods were used. Results: HRGPcreGjd2fl/fl mice had a selective reduction in Cx36 protein in the outer plexiform layer; no reduction in Cx36 was observed in the inner plexiform layer. Cx36 disruption had no effect on the number of cones, the thickness of the photoreceptor layer, or the scotopic ERG responses. However, there was a reduction of the photopic ERG circadian rhythm, with b-wave amplitudes in the day and the night locked in the daytime, light-adapted state. In HRGPcreGjd2+/+and Gjd2fl/fl controls, the circadian rhythm of light-adapted ERG persisted, similar to that in wild type mice. Conclusions: Cx36 regulation contributes to the circadian rhythm of light-adapted ERG; in the absence of photoreceptor gap junctions, mice appear to be in a fully light-adapted state regardless of the time of day. The higher amplitudes and reduced circadian regulation of the b-wave of HRGPcreGjd2fl/fl mice may be due to increased synaptic strength at the cone to ON bipolar cell synapse due to electrotonic isolation of the terminals lacking gap junctions.

Targeted monitoring of patients at high risk of post-transplant lymphoproliferative disease by quantitative Epstein-Barr virus polymerase chain reaction.

BACKGROUND: Epstein-Barr virus (EBV)-associated post-transplantation lymphoproliferative disease (PTLD) is a serious complication after allogeneic stem cell transplantation (SCT). The likelihood of PTLD is increased in the presence of specific risk factors. Monitoring of EBV DNA load and early administration of rituximab in patients with high EBV loads is recommended for high-risk patients. METHODS: Patients at high risk of EBV-associated PTLD were defined as those showing an EBV serological mismatch between donor and recipient, those with lymphoma, those given cord blood grafts, and those with primary EBV disease before SCT. High-risk patients were prospectively monitored by weekly measurement of EBV DNA by quantitative polymerase chain reaction assay, and rituximab was given when the EBV load reached 10,000 copies/mL or symptoms were suggestive of EBV disease. During the study period (July 2005 to the end of June 2007) 131 patients underwent SCT, of whom 53 had high risk factors. A historical control group transplanted between January 2003 to the end of June 2005 was retrospectively used to evaluate the effect of the prospective monitoring strategy. RESULTS: Of the patients, 30% were positive for EBV DNA at least once; 10% of patients with EBV DNAemia developed PTLD. Risk factors of EBV DNAemia were younger age (P=0.04), receiving transplants from mismatched family or unrelated donors (P=0.01), and acute graft-versus-host disease grades II-IV (P=0.001). The overall frequency of PTLD was 3%; 5.7% in the high-risk group and 1.3% in the standard-risk group. Previous splenectomy (P=0.046) was the only significant risk factor associated with PTLD. In the control group, 6 of 150 patients (4%) developed PTLD; 5/53 (9.4%) in the high-risk group and 1/97 (1%) in the standard-risk group. Human leukocyte antigen-mismatched donors (P<0.01) and EBV-positive donors/EBV-negative recipients (P=0.01) had a significant impact on the risk of PTLD. CONCLUSION: A targeted monitoring strategy among patients at a high risk of EBV-associated PTLD might be helpful to decrease the risk of development of PTLD. However, larger prospective studies are needed to verify this hypothesis.

Biological nitrogen and phosphorus removal in UCT-type MBR process.

A lab-scale UCT-type membrane bio-reactor (MBR) was operated for biological nitrogen (N) and phosphorus (P) removal simultaneously. In order to examine biological nutrient removal (BNR) characteristics of MBR, the lab unit was fed with a synthetic strong and weak wastewater. With strong wastewater, a simultaneous removal of N and P was achieved while application of weak wastewater resulted in a decrease of both N and P removal. Recycled nitrate due to the limited organic in weak wastewater operation probably caused a nitrate inhibition in anaerobic zone. In step feed modification with weak wastewater, both N and P removal capability recovered in the system, indicating that the allocation of COD for denitrification at anoxic zone was a key to increase the biological P removal. In addition, the analysis on the specific P uptake rate in anoxic zone demonstrated that denitrifying phosphorus accumulating organism (dPAO) played an important role to remove up to 40% of P along with N. The sludge production characteristics of UCT-type MBR were similar to ordinary activated sludge with BNR capability.

Quantification of adenovirus DNA in unrelated donor hematopoietic stem cell transplant recipients.

BACKGROUND: Adenovirus (AdV) infection is a life threatening condition in immunosuppressed patients. Quantitative AdV assays can improve the clinical management of these patients. OBJECTIVES: To evaluate quantitative measurement of AdV DNA with PCR in blood from hematopoietic stem cell transplant (HSCT) recipients. STUDY DESIGN: Quantitative PCR was used to measure viral DNA levels of AdV in consecutive blood samples from 40 HSCT recipients (27 adults and 13 children) during a 1-year post-engraftment period. All patients received grafts from unrelated donors and were given anti-T-cell antibodies in the conditioning regimen. RESULTS: In the group of 40 patients, six (15%) had detectable AdV DNA in blood for different lengths of time. None of these six patients suffered from severe graft-versus-host disease. In three of the patients a high AdV viral load (>10,000 copies/mL) was detected, one of whom also had high viral load of EBV and CMV and one of EBV only. These three patients died within 2 months after detection of ADV viremia. A low AdV viral load (<500 copies/mL) was detected in three surviving patients and they did not have concomitant high viral load of neither CMV nor EBV. CONCLUSIONS: AdV viremia was present in 15% of the HSCT recipients and a high AdV viral load was associated with fatal outcome. Screening for AdV DNA with quantitative PCR in blood may be of clinical importance in allogeneic HSCT recipients in order to prevent severe clinical virological complications.

Effects of nitrate on the UV photolysis of H2O2 for VOCs degradation in an aqueous solution.

The major objective of this study was to delineate the effect of nitrate on the UV oxidation of benzene and toluene, dissolved in less than 100 microg l(-1), by conducting a bench-scale operation at various reaction times and with various initial concentrations of H2O2 and NO3-. The oxidation of benzene and toluene can be expected to be only about 10% and 18%, respectively, through the photolysis of H2O2 (initial conc. of 50 mg l(-1)), where the reactor was operated at a reaction time of 2 min, with an initial NO(3-)-N concentration of 5 mg l(-1). Nitrate clearly hindered UV oxidation when the initial H2O2 concentration in the reactor was less than 50 mg l(-1). Even if approximately 40% removal could be achieved under the conditions mentioned above (an initial H2O2 concentration of 200 mg l(-1) at a reaction time of 9 min, with a high UV dose), the operating conditions for the 40% removal might be beyond the practical limits applied for effluents discharged from wastewater treatment plants. The results of the experiment also indicate that benzene and toluene can be oxidized in very limited amounts through direct photolysis, without additional oxidation by hydroxyl radicals.

Corneal Epithelial Cells Exhibit Myeloid Characteristics and Present Antigen via MHC Class II.

Purpose: To explore the impact of ocular surface insults on the immunomodulatory capacity and phenotype of corneal epithelial cells (CECs) with a focus on epithelial-mesenchymal transition (EMT). Methods: Corneas were harvested from mice 6 days following scratch injury, ragweed pollen-induced allergy, or herpes simplex virus type 1 (HSV-1) infection and compared to healthy tissue controls. Corneas were enzymatically digested and CECs phenotypically characterized using flow cytometry. CECs were defined as epithelial cell adhesion molecule (EpCAM)-positive CD45-negative cells. CECs were assessed by PCR to evaluate EMT-associated transcripts. Recombinant HSV-1 and transgenic mice were utilized to investigate the role of vascular endothelial growth factor A (VEGFA) on the phenotype observed. The immunomodulatory potential of CECs was assessed in coculture assays with ovalbumin-specific CD4 T cells. Results: Ectopic expression of classic "myeloid" antigens Ly6G, CCR2, and CX3CR1 was identified in CEC subsets from all groups with evidence supporting an underlying partial EMT event resulting from loss of cell-cell contacts. Corneal HSV-1 infection induced Ly6C expression and major histocompatibility complex (MHC)-II upregulation in CECs through a VEGFA-linked mechanism. These Ly6C+ MHC-II+ CECs were found to function as amateur antigen-presenting cells and induced CD4 T cell proliferation in vitro. Conclusions: This study characterizes a novel immunomodulatory CEC phenotype with possible implications for immune privilege, chronic inflammation, and tissue fibrosis. Moreover, the identification of CECs masquerading with multiple "myeloid" antigens warrants careful evaluation of flow cytometry data involving corneal digests.