Flow cytometric quantitation of EpCAM-positive extracellular vesicles by immunomagnetic separation and phospholipid staining method.

Extracellular vesicles (EV) have attracted attention as circulating biomarkers for many diseases, particularly cancer. Conventional immunofluorescence staining has been used for the detection of target antigens on EV by flow cytometry. However, the staining intensity depends on the amount of antigen expressed on the vesicles and is often only around the noise level. Instead of immunofluorescence, we combined immunomagnetic separation using nanosize MACS® MicroBeads with phospholipid staining of EV (IMS-PS method). EpCAM-positive EV were prepared from the culture supernatants of OVCAR3 (EpCAM-high), A431 (EpCAM-low) or Colon-26 (non-human control) cells as cancer models and were examined by the IMS-PS method using EpCAM mAb-coated MicroBeads. By employing Polaric-500c6F as the dye for staining EV phospholipids and using appropriate flow cytometry settings, autofluorescence was excluded, whereas pretreatment of the MicroBeads with conventional blocking agents reduced nonspecific binding to non-target vesicles. These modifications resulted in a linear relation between the number of EV detected and the sample volume, regardless of the level of EpCAM expression on the vesicles. A431 EV spiked into healthy volunteer plasma were enumerated with good accuracy. The IMS-PS method may be useful for clinical evaluation of EV with low levels of antigen expression that are difficult to detect by conventional immunofluorescence.

Synthesis and Evaluation of a 1,3a,6a-Triazapentalene (TAP)-Bonded System.

A method of synthesizing a directly connected 1,3a,6a-triazapentalene (TAP) ring system as a linearly bonded aromatic system with a planar form was established. Various TAP-dimers and a 2-alkyl-TAP-trimer were synthesized and their fluorescence properties were evaluated. Although the direct connection of the TAP ring with other TAP rings did not affect the fluorescence properties in diluted solvent, TAP-dimers showed unique fluorescence properties derived from the aggregation state under highly concentrated conditions. In particular, TAP-dimer 5 f showed aggregation-induced emission in highly concentrated solution, and 5 b showed typical mechanochromic fluorescence in the solid state despite their compact molecular size.

Cardiotocography use for fetal assessment during labor in low- and middle-income countries: A scoping review.

BACKGROUND: The use of cardiotocography (CTG) to improve neonatal outcomes is controversial. The medical settings, subjects, utilizations, and interpretation guidelines of CTG are unclear for low- and middle-income countries (LMICs). OBJECTIVES: To assess and review CTG use for studies identified in LMICs and provide insights on the potential for effective use of CTG to improve maternal and neonatal outcomes. SEARCH STRATEGY: The databases Medline, CINAHL, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) were searched for published and unpublished literature through September 2023. SELECTION CRITERIA: Publications were identified which were conducted in LMICs, based on the World Bank list of economies for 2019; targeting pregnant women in childbirth; and focusing on the utilization of CTG and neonatal outcomes. DATA COLLECTION AND ANALYSIS: Publications were screened, and duplicates were removed. A scoping review was conducted using PRISMA-ScR guidelines. RESULTS: The searches generated 1157 hits, of which 67 studies were included in the review. In the studies there was considerable variation and ambiguity regarding the study settings, target populations, utilizations, timing, frequency, and duration of CTG. While cesarean section rates were extensively investigated as an outcome of studies of CTG itself and the effect of additional techniques on CTG, other clinically significant outcomes, including neonatal mortality, were not well reported. CONCLUSIONS: Variations and ambiguities were found in the use of CTG in LMICs. Due to the limited amount of evidence, studies are needed to examine CTG availability in the context of LMICs.

The short and long-term efficacy of nurse-led interventions for improving blood pressure control in people with hypertension in primary care settings: a systematic review and meta-analysis.

BACKGROUND: Previous systematic reviews suggest that nurse-led interventions improve short-term blood pressure (BP) control for people with hypertension. However, the long-term effects, adverse events, and appropriate target BP level are unclear. This study aimed to evaluate the long-term efficacy and safety of nurse-led interventions. METHODS: We conducted a systematic review and meta-analysis. We searched the Cochrane Central Register of Controlled Trials, PubMed, and CINAHL, as well as three Japanese article databases, as relevant randomized controlled trials from the oldest possible to March 2021. This search was conducted on 17 April 2021. We did an update search on 17 October 2023. We included studies on adults aged 18 years or older with hypertension. The treatments of interest were community-based nurse-led BP control interventions in addition to primary physician-provided care as usual. The comparator was usual care only. Primary outcomes were long-term achievement of BP control goals and serious adverse events (range: 27 weeks to 3 years). Secondary outcomes were short-term achievement of BP control goals and serious adverse events (range: 4 to 26 weeks), change of systolic and diastolic BP from baseline, medication adherence, incidence of hypertensive complications, and total mortality. RESULTS: We included 35 studies. Nurse-led interventions improved long-term BP control (RR 1.10, 95%CI 1.03 to 1.18). However, no significant differences were found in the short-term effects of nurse-led intervention compared to usual care about BP targets. Little information on serious adverse events was available. There was no difference in mortality at both terms between the two groups. Establishing the appropriate target BP from the extant trials was impossible. CONCLUSIONS: Nurse-led interventions may be more effective than usual care for achieving BP control at long-term follow-up. It is important to continue lifestyle modification for people with hypertension. We must pay attention to adverse events, and more studies examining appropriate BP targets are needed. Nurse-led care represents an important complement to primary physician-led usual care.

Current Status and Issues of Collaboration Between Physicians and Pharmacists in Fertility Preservation.

Purpose: This study addresses the critical issue of fertility preservation among young patients with cancer in Japan, recognizing the brief decision-making window and the need for comprehensive support. Pharmacists, well-versed in the side effects of anticancer drugs, can play a vital role in this support process. However, the extent of pharmacists' involvement in fertility preservation remains unclear. We aimed to investigate pharmacists' roles in addressing cancer treatment-induced fertility concerns and their collaboration with physicians, offering insights into enhancing pharmacist participation in fertility preservation. Methods: A survey conducted between April and July 2022 targeted doctors and pharmacists at cancer treatment hospitals, along with pharmacists affiliated with the Japanese Society of Pharmaceutical Health Care and Sciences. Results: Our findings indicated that although pharmacists had limited knowledge about gonadotoxicity and fertility, they expressed readiness to conduct research and provide information when consulted. Approximately 10%-20% of the pharmacists participated in explaining the primary disease at diagnosis. Pharmacists played a more prominent role after establishing chemotherapy regimens, with less involvement in its formulation. Notably, treatment decision case conferences emerged as crucial forums for gathering patient data, confirming treatment plans, and identifying those in need for fertility preservation information. Roughly half of the pharmacists attended these conferences, suggesting a need for increased participation. Conclusion: Enhancing physician-pharmacist collaboration could be pivotal for effective fertility preservation. This requires augmenting the knowledge and awareness of both professions and encouraging greater participation in case conferences to create a conducive environment for addressing this critical aspect of cancer care.

Components and Delivery Formats of Cognitive Behavioral Therapy for Chronic Insomnia in Adults: A Systematic Review and Component Network Meta-Analysis.

Importance: Chronic insomnia disorder is highly prevalent, disabling, and costly. Cognitive behavioral therapy for insomnia (CBT-I), comprising various educational, cognitive, and behavioral strategies delivered in various formats, is the recommended first-line treatment, but the effect of each component and delivery method remains unclear. Objective: To examine the association of each component and delivery format of CBT-I with outcomes. Data Sources: PubMed, Cochrane Central Register of Controlled Trials, PsycInfo, and International Clinical Trials Registry Platform from database inception to July 21, 2023. Study Selection: Published randomized clinical trials comparing any form of CBT-I against another or a control condition for chronic insomnia disorder in adults aged 18 years and older. Insomnia both with and without comorbidities was included. Concomitant treatments were allowed if equally distributed among arms. Data Extraction and Synthesis: Two independent reviewers identified components, extracted data, and assessed trial quality. Random-effects component network meta-analyses were performed. Main Outcomes and Measures: The primary outcome was treatment efficacy (remission defined as reaching a satisfactory state) posttreatment. Secondary outcomes included all-cause dropout, self-reported sleep continuity, and long-term remission. Results: A total of 241 trials were identified including 31 452 participants (mean [SD] age, 45.4 [16.6] years; 21 048 of 31 452 [67%] women). Results suggested that critical components of CBT-I are cognitive restructuring (remission incremental odds ratio [iOR], 1.68; 95% CI, 1.28-2.20) third-wave components (iOR, 1.49; 95% CI, 1.10-2.03), sleep restriction (iOR, 1.49; 95% CI, 1.04-2.13), and stimulus control (iOR, 1.43; 95% CI, 1.00-2.05). Sleep hygiene education was not essential (iOR, 1.01; 95% CI, 0.77-1.32), and relaxation procedures were found to be potentially counterproductive(iOR, 0.81; 95% CI, 0.64-1.02). In-person therapist-led programs were most beneficial (iOR, 1.83; 95% CI, 1.19-2.81). Cognitive restructuring, third-wave components, and in-person delivery were mainly associated with improved subjective sleep quality. Sleep restriction was associated with improved subjective sleep quality, sleep efficiency, and wake after sleep onset, and stimulus control with improved subjective sleep quality, sleep efficiency, and sleep latency. The most efficacious combination-consisting of cognitive restructuring, third wave, sleep restriction, and stimulus control in the in-person format-compared with in-person psychoeducation, was associated with an increase in the remission rate by a risk difference of 0.33 (95% CI, 0.23-0.43) and a number needed to treat of 3.0 (95% CI, 2.3-4.3), given the median observed control event rate of 0.14. Conclusions and Relevance: The findings suggest that beneficial CBT-I packages may include cognitive restructuring, third-wave components, sleep restriction, stimulus control, and in-person delivery but not relaxation. However, potential undetected interactions could undermine the conclusions. Further large-scale, well-designed trials are warranted to confirm the contribution of different treatment components in CBT-I.

The Role of Autophagy in the Female Reproduction System: For Beginners to Experts in This Field.

Autophagy is a fundamental process involved in regulating cellular homeostasis. Autophagy has been classically discovered as a cellular process that degrades cytoplasmic components non-selectively to produce energy. Over the past few decades, this process has been shown to work in energy production, as well as in the reduction of excessive proteins, damaged organelles, and membrane trafficking. It contributes to many human diseases, such as neurodegenerative diseases, carcinogenesis, diabetes mellitus, development, longevity, and reproduction. In this review, we provide important information for interpreting results related to autophagic experiments and present the role of autophagy in this field.

Risk factors associated with fatal pulmonary hemorrhage in locally advanced non-small cell lung cancer treated with chemoradiotherapy.

BACKGROUND: The purpose of this study was to identify the risk factors associated with fatal pulmonary hemorrhage (PH) in patients with locally advanced non-small cell lung cancer (NSCLC), treated with chemoradiotherapy. METHODS: The medical records of 583 patients with locally advanced NSCLC, who were treated with chemoradiotherapy between July 1992 and December 2009 were reviewed. Fatal PH was defined as PH leading to death within 24 h of its onset. Tumor cavitation size was defined by the cavitation diameter/tumor diameter ratio and was classified as minimum (< 0.25), minor (≥ 0.25, but < 0.5), and major (≥ 0.5). RESULTS: Of the 583 patients, 2.1% suffered a fatal PH. The numbers of patients with minimum, minor, and major cavitations were 13, 11, and 14, respectively. Among the 38 patients with tumor cavitation, all 3 patients who developed fatal PH had major cavitations. On multivariate analysis, the presence of baseline major cavitation (odds ratio, 17.878), and a squamous cell histology (odds ratio, 5.491) proved to be independent significant risk factors for fatal PH. Interestingly, all patients with fatal PH and baseline major cavitation were found to have tumors with squamous cell histology, and the occurrence of fatal PH in patients having both risk factors was 33.3%. CONCLUSIONS: Patients at high risk of fatal PH could be identified using a combination of independent risk factors.

Primary palliative care in Japan: needs estimation and projections - national database study with international comparisons.

OBJECTIVES: We aimed to estimate the potential population that requires palliative care, clarify the relationship between this population and the rate of ageing in Japan, and compare these trends with those of other countries. DESIGN: We used the national death registration data and population projections for Japan to estimate the population in need of palliative care using the minimal estimate method developed by Murtagh et al. Linear regression was used to create a model of mortality using sex, age at intervals of 5 years, and each major disease classification. We calculated the future population in need of palliative care until 2040 and compared the ageing data to those of other countries. SETTING/PARTICIPANTS: All adults in Japan who died from 1980 to 2040 at intervals of 5 years. RESULTS: The number of people who might need palliative care from 2020 to 2040 will also increase linearly from 1 059 000 to 1 405 000. The proportion of Alzheimer's, dementia and senility of the total need for palliative care will increase to 43.4% in 2040. The correlation coefficient between the proportion of the population in need of palliative care and the rate of ageing was 0.24 in developed countries. CONCLUSION: In Japan, the population requiring palliative care in 2040 will be 1.5 times that in 2015. Palliative care needs to be provided urgently for people with Alzheimer's disease, dementia and senility. The proportion of patients in need of palliative care may not change, although the number of patients requiring such gradually increases in developed countries.

Non-pharmacological interventions for cancer-related fatigue in terminal cancer patients: a systematic review and meta-analysis.

BACKGROUND: Cancer-related fatigue (CRF) is a highly distressing symptom in patients with cancer. Although various interventions have been reported to reduce fatigue, few are available for use in terminally ill cancer patients, and it is unknown which interventions are effective. They are also often difficult to implement in terminally ill patients with cancer. We, therefore, assessed the recommended interventions to reduce CRF in terminally ill cancer patients. METHODS: Four electronic databases were searched to identify studies published between January 2015 and March 2021. The inclusion criteria were terminally ill cancer patients; non-pharmacological interventions; studies in which usual care or control groups were compared, or comparisons were made prior to the post-intervention period; studies in which the primary outcome was fatigue scale or symptom scale (including those measuring fatigue on a subscale); and experimental study designs including randomised controlled trials (RCTs) and quasi-experimental studies. A summary of the data extracted from each study was created. We also conducted a meta-analysis of the RCTs. RESULTS: A total of 1,954 publications were identified from the initial database, eight of which were included in this study. Three RCTs and five non-RCTs were included in the final evaluation. Most of the studies had a small number of participants. We conducted a meta-analysis of two of the three RCTs included in this study. There was insufficient evidence to determine the effects of the interventions compared to the controls [standard mean difference, -0.05; 95% confidence interval (CI): -0.48 to 0.37; two studies; 290 participants; I2=65%]. CONCLUSIONS: Few reports exist on non-pharmacological interventions for patients with terminal cancer and there was insufficient evidence to determine the effect of the interventions on fatigue. This highlights the lack of RCTs on non-pharmacological procedures and therapies for reducing fatigue.

Pilot plant demonstration of temperature impacts on the methanogenic performance and membrane fouling control of the anaerobic membrane bioreactor in treating real municipal wastewater.

A 5,000-L anaerobic membrane bioreactor (AnMBR) fed with actual municipal wastewater was employed to study the impact of temperature drops on methanogenic performance and membrane fouling. With temperature dropped from 25 °C to 15 °C, the methane yield decreased from 0.244 to 0.205 NL-CH4/g-CODremoval and the dissolved methane increased from 29% to 43%, resulted in the methanogenic performance reduced by 25%. The membrane rejection offset the deteriorated anaerobic digestion at low temperatures and ensured the stable COD removal efficiency of 84.5%-90.0%. The synergistic effects of the increased microbial products and viscosity and the residual inorganic foulants aggravated the membrane fouling at lower temperatures. As the organic fouling was easily removed by NaClO, the inorganics related to the elements of S, Ca and Fe were the stubborn membrane foulants and required the enhanced acid membrane cleaning. These findings obtained under the quasi-practical condition are expected to promote the practical applications of mainstream AnMBR.

Assessment of blinding in randomized controlled trials of antidepressants for depressive disorders 2000-2020: A systematic review and meta-analysis.

Background: In double-blind randomized controlled trials (RCTs) of antidepressants, blinding can be broken due to the apparent side effects, and unsuccessful blinding can lead to overestimation of effect sizes. New generation antidepressants with less severe side effects may be less susceptible to broken blinding. However, successfulness of blinding in new generation antidepressant trials and its influence on trial effect size estimates remain unclear. Methods: Extending a previous systematic review assessing blinding successfulness in psychiatric trials (2000-2010), we searched PubMed/Medline for double-blinded antidepressant RCTs (2010-2020) for trials assessing blinding success. Our primary outcome was the degree of blinding successfulness, measured as kappa statistics between guesses and true allocations. We used random-effects meta-analysis to synthesize studies. We used meta-regression and Pearson's r to examine the relationship between blinding success and effect sizes. This study is registered with PROSPERO (CRD42021249973). Findings: Among 154 eligible studies, 11 (7·1%) contained information on blinding assessment between 2010 and 2020. Five studies were added from the previous review, and altogether nine of the 16 studies provided usable data. Agreement in individual studies ranged from κ=-0·14 to 0·38. The summary agreement between guesses and the truth was 0·21 (95% CI: 0·14 to 0·28) among patients and 0·17 (95% CI: 0·05 to 0·30) among assessors. Blinding success was not associated with effect size (patients: r = 0·37, p = 0·32; assessors: r = 0·28; p = 0·72). Meta-regression also failed to find a significant relationship between blinding success and depression effect sizes (ß=0·06, p = 0·09). Interpretation: Less than 10% of the antidepressant RCTs reported blinding assessment. The results in new generation antidepressant trials indicated that patients and assessors were unlikely to be able to judge treatment allocation. There was little evidence that the extent of unblinding biased the effect size estimates of new generation antidepressants. Funding: None.

Japanese Version of the Mobile App Rating Scale (MARS): Development and Validation.

BACKGROUND: The number of mobile health (mHealth) apps continues to rise each year. Widespread use of the Mobile App Rating Scale (MARS) has allowed objective and multidimensional evaluation of the quality of these apps. However, no Japanese version of MARS has been made available to date. OBJECTIVE: The purposes of this study were (1) to develop a Japanese version of MARS and (2) to assess the translated version's reliability and validity in evaluating mHealth apps. METHODS: To develop the Japanese version of MARS, cross-cultural adaptation was used using a universalist approach. A total of 50 mental health apps were evaluated by 2 independent raters. Internal consistency and interrater reliability were then calculated. Convergent and divergent validity were assessed using multitrait scaling analysis and concurrent validity. RESULTS: After cross-cultural adaptation, all 23 items from the original MARS were included in the Japanese version. Following translation, back-translation, and review by the author of the original MARS, a Japanese version of MARS was finalized. Internal consistency was acceptable by all subscales of objective and subjective quality (Cronbach α=.78-.89). Interrater reliability was deemed acceptable, with the intraclass correlation coefficient (ICC) ranging from 0.61 to 0.79 for all subscales, except for "functionality," which had an ICC of 0.40. Convergent/divergent validity and concurrent validity were also considered acceptable. The rate of missing responses was high in several items in the "information" subscale. CONCLUSIONS: A Japanese version of MARS was developed and shown to be reliable and valid to a degree that was comparable to the original MARS. This Japanese version of MARS can be used as a standard to evaluate the quality and credibility of mHealth apps.

The first pilot-scale demonstration of the partial nitritation/anammox-hydroxyapatite process to treat the effluent of the anaerobic membrane bioreactor fed with the actual municipal wastewater.

Recently, it was reported that for synthetic low-strength wastewater, the excellent nitrogen removal rate (NRR) accompanied with phosphorus removal could be achieved through the partial nitritation/anammox (PNA)-hydroxyapatite (HAP) process. Thus, this research further investigated the performance of the pilot-scale PNA-HAP process treating the effluent of an anaerobic membrane bioreactor (AnMBR) fed with the actual municipal wastewater. The results showed that with the hydraulic retention time of 4.0 h, the influent ammonium concentration ranging from 36.0 to 41.0 mg/L, and the BOD5 ranging from 6.3 to 12.7 mg/L, the average NRR and the nitrogen removal efficiency was 0.13 kg/m3/d and 63.38%, respectively. The specific activity test of sludge confirmed that the PNA process was the main nitrogen metabolism pathway. The effluent nitrate and the BOD5 were almost zero, indicating the existence of denitrification activity in reactor. Given that the oxygenation condition, the heterotrophic organic matter oxidization activity also occurred in reactor. The sludge analysis confirmed the phosphate formation in sludge. Thus, in the reactor, four kinds of biological activities and chemical crystallization occurred harmoniously in sludge. From the mixed liquid volatile suspended solid of 2.4 g/L and the low distribution range of granule size, it was obvious that the sludge had a high dispersity. Based on the well settling ability of sludge during the operation, it was inferred that there was a close bond between biomass and HAP in sludge, which was helpful to enhance the settleability of sludge granule. Besides, the phosphorus-containing sludge was suitable as the fertilizer. In all, this study demonstrated that the PNA-HAP process is an ideal alternative treating the effluent of the AnMBR process in the municipal wastewater treatment.

Development and validation of the Japanese version of the uMARS (user version of the mobile app rating system).

BACKGROUND: Although the global market of Mobile Health Apps (mHealth apps) continues to grow dramatically, most mHealth apps still not only lack evidence base but have even not been evaluated for the basic usability or functionality. The User Version of the Mobile App Rating Scale (uMARS) was developed to allow end users to assess mHealth apps objectively and subjectively. However, there is no Japanese version of uMARS to date. OBJECTIVE: The purpose of this study is (1) to develop a validated Japanese version of uMARS and (2) to assess the translated version's reliability and validity in evaluating mHealth apps. METHODS: The original uMARS was adapted for Japanese use by four specialists using universalist cross-cultural methods. Translation/back-translation was reviewed by the author of the original version of uMARS, and confirmed. Its reliability and validity were further evaluated as part of a prospective cohort study of postoperative patients using a new mHealth app. RESULTS: Conceptual equivalence was analyzed and all items in all subcategories of the original uMARS were included in the Japanese version. Internal consistency was deemed acceptable for all subscales of objective and subjective quality with a Cronbach's alpha of 0.75-0.85. Test-retest reliability of all subscales was also acceptable with intraclass correlation coefficients (ICCs) of 0.57-0.88. Convergent/divergent validity and concurrent validity were also considered acceptable. CONCLUSION: A Japanese version of uMARS was cross-culturally validated and found to be as reliable as the original uMARS. This Japanese version of uMARS is expected to become a standard tool in assessing the quality of mHealth apps in Japan.

Seasonal temperatures impact on the mass flows in the innovative integrated process of anaerobic membrane bioreactor and one-stage partial nitritation-anammox for the treatment of municipal wastewater.

A pilot-scale anaerobic membrane bioreactor (AnMBR) integrated with a one-stage partial nitritation-anammox (PN/A) reactor was operated for the treatment of municipal wastewater (MWW) at seasonal temperatures of 15-25 °C. The removal efficiencies of COD and total nitrogen (TN) were always > 90% and > 75% respectively. The methanogenesis and PN/A were identified as the primary removal pathways of COD and TN, respectively, and were suppressed at low temperatures. With the temperature dropped from 25 °C to 20 °C to 15 °C, the methane-accounted COD decreased from 63.1% to 59.6% to 48.4%, and the PN/A-accounted TN decreased from 58.1% to 51.7% to 45.3%. The AnMBR and PN/A mutually complement each other in this combined process, as the AnMBR removed 8.5%-16.1% of TN by sludge entrainment and the PN/A reactor removed 2.6%-3.4% of COD by denitrification and aerobic oxidation. These results highlighted the strong feasibility of applying the AnMBR-PN/A process to the treatment of MWW in temperate climate.

Treating the filtrate of mainstream anaerobic membrane bioreactor with the pilot-scale sludge-type one-stage partial nitritation/anammox process operated from 25 to 15 °C.

At ambient temperature condition, the one-stage partial nitritation/anammox (PNA) process has been successfully adopted to treat the filtrate from the mainstream anaerobic membrane bioreactor (AnMBR). However, there is no investigation of the performance of this process at low-temperature condition. In this study, the nitrogen removal performance of a pilot-scale PNA reactor at the temperature of 15 °C for treating the filtrate of a mainstream AnMBR was investigated. The nitrogen removal rate of 0.09 kg/m3/d and the nitrogen removal efficiency of 37.6% were achieved. The anammox reaction was the rate-limiting step of the nitrogen removal. Nitrogen removal was attributed in part to denitrification activity. The microbial community analysis confirmed that the main functional bacteria comprised of genus Nitrosomonas and genus Kuenenia. In sum, this research demonstrated the applicability of PNA process for mainstream AnMBR filtrate treatment to some extent and enriched the related knowledge.

Biomass retention and microbial segregation to offset the impacts of seasonal temperatures for a pilot-scale integrated fixed-film activated sludge partial nitritation-anammox (IFAS-PN/A) treating anaerobically pretreated municipal wastewater.

Partial nitritation-anammox (PN/A) is a promising deammonification process to develop energy-neutral wastewater treatment plants. However, the mainstream application of PN/A still faces the challenges of low nitrogen concentration and low temperatures, and has not been studied under a realistic condition of large-scale reactor (kiloliter level), real municipal wastewater (MWW) and seasonal temperatures. In this research, a pilot-scale one-stage PN/A, with integrated fixed-film activated sludge (IFAS) configuration, was operated to treat the real MWW pretreated by anaerobic membrane bioreactor. The removal efficiency of total nitrogen (TN) was 79.4%, 75.7% and 65.9% at 25, 20 and 15°C, corresponding to the effluent TN of 7.3, 9.7 and 12.0 mg/L, respectively. The suppression of ammonium-oxidizing bacteria (AOB) and anammox bacteria (AnAOB) occurred at lower temperatures, and the significant decrease in AOB treatment capacity was the reason for the poorer nitrogen removal at 15°C. Biomass retention and microbial segregation were successfully achieved. Specifically, Candidatus_Brocadia and Candidatus_Kuenenia were main AnAOB genera and mainly enriched on carriers, Nitrosomonas and uncultured f_Chitinophagaceae were main AOB genera and mainly distributed in suspended sludge and retained by sedimentation tank. Moreover, nitrite-oxidizing bacteria (NOB) were sufficiently suppressed by intermittent aeration and low dissolved oxygen, the presence of heterotrophic bacteria upgraded the PN/A to a simultaneous partial nitritation, anammox, denitrification, and COD oxidation (SNADCO) system, which improved the overall removal of TN and COD. The results of this investigation clearly evidence the strong feasibility of PN/A as a mainstream nitrogen removal process in temperate climates.

Bone morphogenetic protein-2 enhances gonadotropin-independent follicular development via sphingosine kinase 1.

PROBLEM: Pre-ovulatory mature follicles are not readily induced from gonadotropin (Gn)-independent early follicles in the poor ovarian response (POR) state, characterized by reduced number of retrieved oocytes. Bone morphogenetic protein (BMP), which is expressed in the ovary, contributes to early folliculogenesis, but its precise underlying mechanism remains unknown. The purpose of this study was to examine the effects of BMP-2 on granulosa cells (GCs) of Gn-independent early follicles. METHOD OF STUDY: Sphingosine kinase 1 (SPHK1) localization, which produces sphingosine 1-phosphate (S1P), was examined in human early follicles by immunohistochemistry. SPHK1 mRNA levels were examined in Gn-independent bovine GCs (bGCs) and human nonluteinized granulosa cell line (HGrC1) cells. Phosphorylated Yes-associated protein (YAP) expression was evaluated by Western blot, and its localization was evaluated immunocytochemically in bGCs. Verteporfin, a selective YAP inhibitor, was used to explore the influence of YAP on BMP-2-induced bGCs proliferation. RESULTS: The expression of SPHK1 was observed in human GCs of primary and secondary follicles. BMP-2 significantly induced SPHK1 mRNA expression in bGCs and HGrC1 cells. Both BMP-2 and S1P decreased phosphorylated YAP protein levels and induced the nuclear translocation of YAP significantly, thereby increasing the number of bGCs by suppressing the Hippo pathway. This BMP-2-induced cell proliferation was completely blocked by verteporfin. CONCLUSION: This is a first report showing that BMP-2 up-regulated SPHK1 mRNA expression in GCs and promoted GCs proliferation through Hippo pathway suppression. Thus, BMP-2 contributes to Gn-independent folliculogenesis via SPHK1, suggesting a potential therapeutic strategy for the POR patients with follicular dysgenesis.

A successful start-up of an anaerobic membrane bioreactor (AnMBR) coupled mainstream partial nitritation-anammox (PN/A) system: A pilot-scale study on in-situ NOB elimination, AnAOB growth kinetics, and mainstream treatment performance.

In this pilot-scale study, an innovative mainstream treatment process that couples the anaerobic membrane reactor (AnMBR) with a one-stage PN/A system was proposed for advancing the concept of carbon neutrality in the municipal wastewater treatment plant. This work demonstrates the start-up procedure of a pilot-scale one-stage PN/A system for mainstream treatment. The 255-day start-up of the one-stage PN/A system involved the cultivation of ammonium-oxidizing bacteria (AOB) from the activated sludge, suppression of nitrite-oxidizing bacteria (NOB), investigation of in-situ growth kinetics of anammox bacteria (AnAOB), and the 50-day operation of the pilot-scale AnMBR-PN/A process for natural mainstream treatment. It is verified in the pilot-scale system for the first time that the in-situ free ammonia (FA) and free nitrous acid (FNA) exposure could effectively eliminate the Nitrospira (the NOB genus) while retaining the Nitosonomas (the AOB genus) community in the suspended sludge. NOB community rebounding was not detected even at the mainstream conditions with low nitrogen concentrations (Influent ammonium concentration=38±6 mg-NH4+-N/L) by intermittent aeration to control the system dissolved oxygen (DO) below 0.5 mg/L. The results of the mainstream treatment showed that the average effluent total nitrogen (TN) in the coupled process was generally lower than 10 mg-N/L, which meets the discharge limits of most prefectures in Japan. The investigated results of the in-situ anammox bacteria (AnAOB) growth kinetics suggested that the promoted start-up strategy of taking advantage of the warm months with higher mainstream temperature to achieve the rapid in-situ growth of the AnAOB is applicable in the investigated regions. From the perspective of the removal performance of the TN and organic substance, the AnMBR-PN/A process has great potential as the layouts of the carbon-neutral mainstream wastewater treatment plants.