Expansion of Anticomplement Therapy Indications from Rare Genetic Disorders to Common Kidney Diseases.

Complement constitutes a major part of the innate immune system. The study of complement in human health has historically focused on infection risks associated with complement protein deficiencies; however, recent interest in the field has focused on overactivation of complement as a cause of immune injury and the development of anticomplement therapies to treat human diseases. The kidneys are particularly sensitive to complement injury, and anticomplement therapies for several kidney diseases have been investigated. Overactivation of complement can result from loss-of-function mutations in complement regulators; gain-of-function mutations in key complement proteins such as C3 and factor B; or autoantibody production, infection, or tissue stresses, such as ischemia and reperfusion, that perturb the balance of complement activation and regulation. Here, we provide a high-level review of the status of anticomplement therapies, with an emphasis on the transition from rare diseases to more common kidney diseases.

MASP3 Deficiency in Mice Reduces but Does Not Abrogate Alternative Pathway Complement Activity Due to Intrinsic Profactor D Activity.

Complement factor D (FD) is a rate-limiting enzyme of the alternative pathway (AP). Recent studies have suggested that it is synthesized as an inactive precursor and that its conversion to enzymatically active FD is catalyzed by mannan-binding lectin-associated serine protease 3 (MASP3). However, whether MASP3 is essential for AP complement activity remains uncertain. It has been shown that Masp1/3 gene knockout did not prevent AP complement overactivation in a factor H-knockout mouse, and a human patient lacking MASP3 still retained AP complement activity. In this study, we have assessed AP complement activity in a Masp3-knockout mouse generated by CRISPR/Cas9 editing of the Masp1/3 gene. We confirmed specific Masp3 gene inactivation by showing intact MASP1 protein expression and absence of mature FD in the mutant mice. Using several assays, including LPS- and zymosan-induced C3b deposition and rabbit RBC lysis tests, we detected plasma concentration-dependent AP complement activity in Masp3 gene-inactivated mice. Thus, although not measurable in 5% plasma, significant AP complement activity was detected in 20-50% plasma of Masp3 gene-inactivated mice. Furthermore, whereas FD gene deletion provided more than 90% protection of CD55/Crry-deficient RBCs from AP complement-mediated extravascular hemolysis, Masp3 gene deletion only provided 30% protection in the same study. We also found pro-FD to possess intrinsic catalytic activity, albeit at a much lower level than mature FD. Our data suggest that MASP3 deficiency reduces but does not abrogate AP complement activity and that this is explained by intrinsic pro-FD activity, which can be physiologically relevant in vivo.

Effects of brief family psychoeducation on family caregiver burden of people with schizophrenia provided by psychiatric visiting nurses: a cluster randomised controlled trial.

BACKGROUND: The purpose of this study was to examine the effects of a brief family psychoeducation (BFP) programme provided by psychiatric visiting nurses on caregiver burden of family caregivers of people with schizophrenia through a cluster randomised controlled trial (cRCT). METHODS: The study was a two-arm, parallel-group cRCT. Forty-seven psychiatric visiting nurse agencies were randomly allocated to the BFP programme group (intervention group) or treatment as usual group (TAU; control group). Caregivers of people with schizophrenia were recruited by psychiatric visiting nurses using a randomly ordered list. The primary outcome was caregiver burden, measured using the Japanese version of the Zarit Burden Interview. Outcome assessments were conducted at baseline, 1-month follow-up, and 6-month follow-up. Intention-to-treat analysis was conducted to examine the effects of the BFP programme on caregiver burden. RESULTS: Thirty-four psychiatric visiting nurse agencies and 83 family caregivers of people with schizophrenia participated in the study. The participant attrition rate was less than 20%. Adherence to the program was 100%. Compared with TAU group, the BFP programme group had decreased caregiver burden. However, this improvement was not significant at 1-month follow-up (adjusted mean difference [aMD] = 0.27, 95% CI = - 5.48 to 6.03, p = 0.93, d = 0.01) or 6-month follow-up (aMD = - 2.12, 95% CI = - 7.80 to 3.56, p = 0.45, d = 0.11). CONCLUSIONS: The BFP programme provided by psychiatric visiting nurses did not achieve significant decreases in caregiver burden. This result may be attributed to the difficulty in continuing the research due to the COVID-19 pandemic, which prevented us from achieving the targeted sample size necessary to meet the statistical power requirements, as well as to the participation of caregivers with relatively low burden. However, the program had the advantage of high adherence to treatment plan. Further studies should be conducted with a larger sample size and a more diverse sample that includes caregivers with a higher care burden. TRIAL REGISTRATION: The study protocol was registered in the University Hospital Medical Information Network Clinical Trials Registry (UMIN000038044) on 2019/09/18.

Rehospitalisation rates after long-term follow-up of patients with severe mental illness admitted for more than one year: a systematic review.

AIMS: This study aimed to conduct a systematic review of studies on the outcomes of long-term hospitalisation of individuals with severe mental illness, considering readmission rates as the primary outcome. METHODS: Studies considered were those in which participants were aged between 18 and 64 years with severe mental illness; exposure to psychiatric hospitals or wards was long-term (more than one year); primary outcomes were readmission rates; secondary outcomes were duration of readmission, employment, schooling, and social participation; and the study design was either observational or interventional with a randomised controlled trial (RCT) design. Relevant studies were searched using MEDLINE, PsycINFO, Web of Science, CINAHL, and the Japan Medical Abstract Society. The final search was conducted on 1 February 2022. The risk of bias in non-randomised studies of interventions was used to assess the methodological quality. A descriptive literature review is also conducted. RESULTS: Of the 11,999 studies initially searched, three cohort studies (2,293 participants) met the eligibility criteria. The risk of bias in these studies was rated as critical or serious. The 1-10 years readmission rate for patients with schizophrenia who had been hospitalised for more than one year ranged from 33 to 55%. The average of readmission durations described in the two studies was 70.5 ± 95.6 days per year (in the case of a 7.5-year follow-up) and 306 ± 399 days (in the case of a 3-8-year follow-up). None of the studies reported other outcomes defined in this study. CONCLUSIONS: The readmission rates in the included studies varied. Differences in the follow-up period or the intensity of community services may have contributed to this variability. In countries preparing to implement de-institutionalisation, highly individualised community support should be designed to avoid relocation to residential services under supervision. The length of stay for readmissions was shorter than that for index admissions. The results also imply that discharge to the community contributes to improved clinical outcomes such as improved social functioning. The validity of retaining patients admitted because of the risk of rehospitalisation was considered low. Future research directions have also been discussed.

Integrated consumers' sustainable and healthy dietary behavior patterns: Associations between demographics, psychological factors, and meal preparation habits among Japanese adults.

The consumers' Sustainable and Healthy Dietary Behaviors (SHDBs) links sustainable behaviors related to eating (e.g., choosing, cooking, eating, and disposing of foods) with the level of "dietary behavior." It was hypothesized that SHDBs will vary depending on consumers' meal preparation habits, such as food selection, cooking frequency, and skill. This study examined (1) some of the behavioral patterns of SHDBs and (2) independent predictors of each pattern. In total, 508 Japanese adults who completed cross-sectional self-administered questionnaire surveys in December 2021 were included in the study. Principal component analysis (PCA) was used to identify the behavioral patterns of the 30-item SHDBs developed previously by the authors. Additionally, independent predictors of each component score were analyzed using multiple linear regression models, which included demographics, psychological factors concerning SHDBs (pros and cons for the decisional balance of SHDBs and number of environmental issues of interest), and meal preparation habits. Participants' median age was 41 (18-59) years (women: n = 259, 51.0%). Five SHDB patterns with several independent predictors were identified: amalgamation of all SHDBs, healthy food choices but unsustainable cooking, sustainable disposal, avoiding plastic products, and sustainable behaviors regarding expiry date. The multiple linear regression analysis demonstrated the independent predictors of the amalgamation of all SHDBs, such as being a man (ß = -0.105), greater pros (ß = 0.134), higher cooking frequency (ß = 0.443) and skill (ß = 0.244). Greater pros and/or smaller cons were associated with four of the five SHDB patterns, including an amalgamation of all SHDBs, healthy food choice and unsustainable cooking, avoiding disposal, and avoiding plastic products. Sustainable and healthy diet education must be implemented to ensure the adoption of all SHDBs.

Sensory profile of picky eaters among college female students.

Picky eaters are at risk of an unbalanced diet, which is critical for women of reproductive age. A sensory profile, which is a potential factor in picky eating, has not been well researched. This study assessed the differences in sensory profile and dietary intake according to the picky eating status among female Japanese undergraduate college students. Cross-sectional data were obtained from the Ochanomizu Health Study conducted in 2018. The questionnaire included items regarding demographic characteristics, picky eating status, sensory profile, and dietary intake. Sensory profile was assessed using the Adult/Adolescent Sensory Profile questionnaire, and dietary intakes were calculated using a brief-type self-administered diet history questionnaire. Among the 111 participants, 23% were picky eaters and 77% were non-picky eaters. The age, body mass index and household status did not differ between the picky eaters and non-picky eaters. Being a picky eater was associated with higher scores on sensory sensitivity and sensation avoiding, and lower thresholds for taste and smell, touch, and auditory stimuli than being a non-picky eater. Of the picky eaters, 58% and 100% were at a high risk for folate and iron deficiencies, respectively, compared to 35% and 81% of non-picky eaters. Nutrition education for picky eaters in reproductive age to increase vegetable dishes comfortably in their diet is suggested to prevent anemia during their future pregnancy.

Mixed-Methods Evaluation of Father Participation in an Adolescent Obesity Prevention Program With Multiple Delivery Methods.

Padres Preparados, Jóvenes Saludables was a Latino family-based obesity prevention intervention implemented from 2017 to 2020 across eight programs in-person only, in a blended format (online/in-person), and online only. The intervention aimed to enhance father parenting skills to improve adolescent diet and activity behaviors. Mothers were encouraged to attend. Factors associated with participation were explored using a mixed-methods, qualitative (focus group/individual interviews by Zoom) and quantitative (process evaluation) design. Eleven focus group and 24 individual interviews were completed after participation with 24 fathers, 27 mothers, and 40 adolescents with responses not sorted by delivery method before analysis. Binomial logistic regression models examined associations between fathers' program completion and predictor variables of delivery characteristics, father demographic characteristics, and family attendance patterns. Parents were married (96% fathers, 76% mothers), had low income, a high school education or less (68% fathers, 81% mothers), and had lived in the United States a mean of 19 years. Parents were motivated to participate to improve health, and to be involved with and improve communication with their child. Common barriers to participation were work and life priorities and programmatic factors including scheduling conflicts and technological issues. Participation was greater for fathers attending sessions in-person compared with online only (OR = 11.6). Fathers were more likely to participate if they attended sessions with family members vs. not attending with family members (OR = 7.2). To maximize participation, findings suggest involving multiple parents/caregivers and adolescents, addressing contextual and programmatic barriers, and promoting benefits of better health and relations with family members.

The rice OsERF101 transcription factor regulates the NLR Xa1-mediated immunity induced by perception of TAL effectors.

Plant nucleotide-binding leucine-rich repeat receptors (NLRs) initiate immune responses by recognizing pathogen effectors. The rice gene Xa1 encodes an NLR with an N-terminal BED domain, and recognizes transcription activator-like (TAL) effectors of Xanthomonas oryzae pv oryzae (Xoo). Our goal here was to elucidate the molecular mechanisms controlling the induction of immunity by Xa1. We used yeast two-hybrid assays to screen for host factors that interact with Xa1 and identified the AP2/ERF-type transcription factor OsERF101/OsRAP2.6. Molecular complementation assays were used to confirm the interactions among Xa1, OsERF101 and two TAL effectors. We created OsERF101-overexpressing and knockout mutant lines in rice and identified genes differentially regulated in these lines, many of which are predicted to be involved in the regulation of response to stimulus. Xa1 interacts in the nucleus with the TAL effectors and OsERF101 via the BED domain. Unexpectedly, both the overexpression and the knockout lines of OsERF101 displayed Xa1-dependent, enhanced resistance to an incompatible Xoo strain. Different sets of genes were up- or downregulated in the overexpression and knockout lines. Our results indicate that OsERF101 regulates the recognition of TAL effectors by Xa1, and functions as a positive regulator of Xa1-mediated immunity. Furthermore, an additional Xa1-mediated immune pathway is negatively regulated by OsERF101.

Multiple stakeholders' perspectives on patient and public involvement in community mental health services research: A qualitative analysis.

BACKGROUND: Patient and public involvement (PPI) has become essential in health research. However, little is known about multiple stakeholders' perspectives on the implementation of PPI in community mental health research settings. The present study aimed to qualitatively analyse multiple stakeholders' views on PPI, including potential concerns, barriers and approaches. METHODS: This study involved conducting focus group interviews and collecting qualitative data from 37 participants in multiple stakeholder groups (patients = 6, caregivers = 5, service providers = 7, government staff = 5 and researchers = 14) in the community mental health field. The data were qualitatively analysed using a data-driven approach that derived domains, themes and subthemes related to perspectives on PPI and to specific challenges and approaches for implementing PPI. RESULTS: The qualitative analysis identified four domains. The 'Positive views and expectations regarding PPI' domain consisted of themes related to supportive views of PPI in a mental health service research setting and improvements in the quality of research and service. The 'General concerns about PPI' domain included themes concerning the need for non-PPI research and tokenism, excessive expectations concerning social changes and use of evidence from PPI research, and heavy burdens resulting from PPI. The 'Specific issues regarding the implementation of PPI' domain consisted of four themes, including academic systems, selection methods (e.g., representativeness and conflict of interest issues), relationship building, and ambiguous PPI criteria. In particular, all stakeholder groups expressed concerns about relational equality during PPI implementation in Japan. The 'Approaches to PPI implementation' domain included themes such as facilitating mutual understanding, creating a tolerant atmosphere, establishing PPI support systems (e.g., training, ethics and human resource matching) and empowering patient organizations. CONCLUSION: The study replicated most of the barriers and approaches to PPI reported by qualitative research in Western counties. However, utilization of evidence produced by PPI research and partnership in the PPI process may be particularly serious issues in Japan. Future PPI studies should carefully address solutions that fit each culture. PATIENT OR PUBLIC CONTRIBUTION: A patient-researcher was involved in all stages of this project, from development of the research topic and the protocol to manuscript preparation.

Feasibility of a remotely delivered program to prevent Latino adolescent obesity in the Midwestern USA.

Adolescent obesity prevention programs focusing on Latino fathers may be useful to address Latino adolescent obesity. Adolescent obesity has become an urgent issue because of the coronavirus disease 2019 pandemic, with limited ability to deliver prevention programs in-person. This study aimed to assess the feasibility of a community-based, adolescent obesity prevention program for Latino father-adolescent dyads delivered remotely, adapted from the in-person Padres Preparados, Jóvenes Saludables program. A quasi-experimental design was used to assess the feasibility of the remotely delivered program based on criteria adapted from other feasibility studies of community-based health promotion programs for Latino families. Father-adolescent dyads were recruited at two sites in a Midwestern state during 2020; mothers were also encouraged to participate. Recruitment met the feasibility criteria (65 families expressed interest between August and October) with 26 families participating in this study. The retention ratio (percentage of participants who completed a post-session survey to those who completed the baseline survey) among fathers was acceptable (77%), while a lower percentage of adolescents were retained (68%). The session attendance ratio (number of sessions attended of the eight total sessions offered) was higher among adolescents (88%) and lower among fathers (68%) compared to the criteria. Satisfaction ratings (≥88%) and completeness of data collection at both baseline and post-session survey (≥76%) were acceptable. Favorable results were obtained for parent outcomes, while adolescent outcomes were not favorably changed. This remotely delivered adolescent obesity prevention program was feasible for Latino fathers; however, additional engagement with adolescents may be needed.

Adolescent obesity became a more urgent issue because of the coronavirus disease 2019 pandemic with less physical activity under lockdown, more food insecure families and limited in-person access to programs. Our study concluded that a remotely delivered version of a community-based, adolescent obesity prevention program for Latino father­adolescent dyads (the Padres Preparados, Jóvenes Saludables program) was feasible based on participation, satisfaction and parent outcomes. Additional engagement in activities to improve adolescent health behaviors may be needed to improve the overall effectiveness of the program.

Predictive Association of Low- and High-Fidelity Supported Employment Programs with Multiple Outcomes in a Real-World Setting: A Prospective Longitudinal Multi-site Study.

PURPOSE: The individual placement and support (IPS) model of supported employment is a leading evidence-based practice in community mental health services. In Japan, individualized supported employment that is highly informed by the philosophy of the IPS model has been implemented. While there is a body of evidence demonstrating the association between program fidelity and the proportion of participants gaining competitive employment, the association between fidelity and a wider set of vocational and individual outcomes has received limited investigation. This study aimed to assess whether high-fidelity individualized supported employment programs were superior to low-fidelity programs in terms of vocational outcomes, preferred job acquisition, and patient-reported outcome measures (PROMs). METHODS: A prospective longitudinal study with 24-month follow-up analyzed 16 individualized supported employment programs. The Japanese version of the individualized Supported Employment Fidelity scale (JiSEF) was used to assess the structural quality of supported employment programs (scores: low-fidelity program, ≤ 90; high-fidelity program, ≥ 91). Job acquisition, work tenure, work earnings, job preference matching (e.g., occupation type, salary, and illness disclosure), and PROMs such as the INSPIRE and WHO-Five Well-being index were compared between groups. RESULTS: There were 75 and 127 participants in the low-fidelity group (k = 6) and high-fidelity group (k = 10), respectively. The high-fidelity group demonstrated better vocational outcomes than the low-fidelity group, i.e., higher competitive job acquisition (71.7% versus 38.7%, respectively, adjusted odds ratio (aOR) = 3.6, p = 0.002), longer work tenure (adjusted mean difference = 140.8, p < 0.001), and better match for illness disclosure preference (92.6% versus 68.0%, respectively, aOR = 5.9, p = 0.003). However, we found no differences between groups in other preference matches or PROM outcomes. CONCLUSION: High-fidelity individualized supported employment programs resulted in good vocational outcomes in a real-world setting. However, enhancing service quality to increase desired job acquisition and improve PROMs will be important in the future. CLINICAL TRIAL REGISTRATION: UMIN000025648.

Influence of paternal general parenting on Latino early adolescents' energy balance-related behaviours and interactions with behaviour-specific parenting practices.

OBJECTIVE: To investigate the influence of general parenting dimensions on adolescents' energy balance-related behaviours (EBRB) and its interactions with behaviour-specific parenting practices based on Darling and Steinberg's contextual model of parenting style. DESIGN: Multiple linear regression analyses and the Hayes PROCESS procedure to analyse self-reported cross-sectional survey data. SETTING: In-person survey. PARTICIPANTS: Latino early adolescents and their fathers (n 225 dyads) recruited using convenience sampling from metropolitan areas of north-central USA. RESULTS: Both paternal parenting dimensions of warmth and autonomy granting were positively associated with adolescents' fruit intake, vegetable intake and physical activity. Coercive control was positively associated with adolescents' sugary drink intake and sweets/salty snack intake. These associations were predominantly mediated by the parenting practices of setting expectations/allowances, role modelling, and managing availability and accessibility for corresponding EBRB. After adjusting for parenting practices, paternal warmth was inversely associated with adolescents' screen time, paternal autonomy was positively associated with sugary drink intake, and both paternal warmth and autonomy granting were positively associated with adolescents' fast food intake. In addition, positive associations between fathers' parenting practices and adolescents' sugary drink intake were significantly stronger among those who perceived high v. low levels of paternal warmth and autonomy granting. CONCLUSIONS: Paternal warmth and autonomy granting showed mixed effects on adolescents' EBRB, and coercive control showed undesirable relationships with adolescents' dietary intake via interactions with behaviour-specific parenting practices. Lifestyle intervention programmes for Latino adolescents need to consider incorporating paternal parenting education components.

Differences in reporting food insecurity and factors associated with differences among Latino fathers and mothers.

BACKGROUND: Food security status has been assessed as a representative score for households; however, different members in the same household may perceive and report food insecurity differently. A high prevalence of food insecurity has been reported among Latino households, therefore understanding differences in reporting food insecurity by Latino father-mother dyads may improve accuracy of assessment and plans to address food insecurity. This study aimed to 1) determine demographic characteristics and/or food-related factors associated with perceptions of food security status among Latino father-mother dyads, and 2) identify factors associated with discordance in perceptions of food insecurity between dyads. METHODS: Baseline data were used from a community-based, youth obesity prevention program among Latino families (n = 106 father-mother dyads). Food security was assessed with a 2-item food insecurity screen. Logistic regression models were used to evaluate associations between reporting food security status and predictor variables for fathers, mothers, and dyad-discordant responses. RESULTS: Food insecurity was reported by 39% of fathers and 55% of mothers. Adjusted odds of reporting food insecurity were significantly higher for fathers perceiving their neighborhood was unsafe vs. safe (OR: 3.7, p < 0.05) and reporting lower vs. higher household income (OR: 3.2, p < 0.05). Adjusted odds of reporting food insecurity were significantly higher for mothers perceiving their neighborhood was unsafe vs. safe (OR: 4.1, p < 0.01) and reporting lower vs. higher home availability of fruit and vegetable (OR: 5.5, p < 0.01). Dyad discordance in reporting food security status occurred in 24% of the dyads. Adjusted odds of dyad discordant reports of food insecurity status were significantly higher for dyads reporting discordant responses regarding previous nutrition education (OR: 3.4, p < 0.05) and higher home fruit and vegetable accessibility (OR: 3.1, p < 0.05) compared to dyads reporting concordant responses. Among the 28 dyads who reported discordant nutrition education participation, 21 reported that fathers had never participated but mothers had participated more than once. CONCLUSIONS: Differential factors were associated with reporting food security among Latino father-mother dyads. Nutrition education for fathers that improves awareness of home food supplies and a better understanding of how food accessibility influences maternal perceptions may improve dyad discordance in reporting household food security.

The leaky pipeline in research grant peer review and funding decisions: challenges and future directions.

The growing literature on gender inequality in academia attests to the challenge that awaits female researchers during their academic careers. However, research has not yet conclusively resolved whether these biases persist during the peer review process of research grant funding and whether they impact respective funding decisions. Whereas many have argued for the existence of gender inequality in grant peer reviews and outcomes, others have demonstrated that gender equality is upheld during these processes. In the present paper, we illustrate how these opinions have come to such opposing conclusions and consider methodological and contextual factors that render these findings inconclusive. More specifically, we argue that a more comprehensive approach is needed to further the debate, encompassing individual and systemic biases as well as more global social barriers. We also argue that examining gender biases during the peer review process of research grant funding poses critical methodological challenges that deserve special attention. We conclude by providing directions for possible future research and more general considerations that may improve grant funding opportunities and career paths for female researchers.

Antibody Inhibition of Properdin Prevents Complement-Mediated Intravascular and Extravascular Hemolysis.

Paroxysmal nocturnal hemoglobinuria (PNH) is a serious blood disorder characterized by dysregulated complement activation on blood cells. Eculizumab, the current standard therapy and a humanized anti-C5 mAb, relieves anemia and thrombosis symptoms of PNH patients by preventing complement-dependent intravascular hemolysis (IVH). However, up to 20% of PNH patients on long-term eculizumab treatment still suffer from significant anemia and are transfusion dependent because of extravascular hemolysis (EVH) of C3-opsonized PNH erythrocytes. In this study, we show that function-blocking anti-properdin (P) mAbs dose-dependently inhibited autologous, complement-mediated hemolysis induced by factor H dysfunction. Furthermore, anti-human P (hP) mAbs potently and dose-dependently inhibited acidified serum-induced hemolysis of PNH erythrocytes (Ham test). In contrast to erythrocytes rescued by anti-C5 mAb, nonlysed PNH erythrocytes rescued by anti-P mAb incurred no activated C3 fragment deposition on their surface. These results suggested that anti-P mAbs may prevent EVH as well as IVH of PNH erythrocytes. To test the in vivo efficacy of anti-hP mAbs in preventing EVH, we generated a P humanized mouse by transgenic expression of hP in P knockout mice (hP-Tg/P-/-). In a murine EVH model, complement-susceptible erythrocytes were completely eliminated within 3 d in control mAb-treated hP-Tg/P-/- mice, whereas such cells were protected and persisted in hP-Tg/P-/- mice treated with an anti-hP mAb. Collectively, these data suggest that anti-P mAbs can inhibit both IVH and EVH mediated by complement and may offer improved efficacy over eculizumab, the current standard therapy for PNH.

Differential contribution of C5aR and C5b-9 pathways to renal thrombic microangiopathy and macrovascular thrombosis in mice carrying an atypical hemolytic syndrome-related factor H mutation.

Atypical hemolytic uremic syndrome (aHUS) is a form of thrombotic microangiopathy (TMA) caused by dysregulated complement activation. Clinically, aHUS is effectively treated by an anti-C5 monoclonal antibody (mAb) but whether the disease is mediated by the C5a receptor (C5aR) or C5b-9 pathway, or both, is unknown. Here we address this in a factor H mutant mouse (FHR/R) which developed complement-mediated TMA as well as macrovascular thrombosis caused by an aHUS-related factor H point mutation (mouse W1206R, corresponding to human W1183R). C5 deficiency and anti-C5 mAb treatment blocked all disease manifestations in FHR/R mice. C5aR1 gene deficiency prevented macrovascular thrombosis in various organs but did not improve survival or reduce renal TMA. Conversely, C6 or C9 deficiency significantly improved survival and markedly diminished renal TMA but did not prevent macrovascular thrombosis. Interestingly, as they aged both FHR/R C6-/- and FHR/R C9-/- mice developed glomerular disease reminiscent of C3 glomerulonephritis. Thus, C5aR and C5b-9 pathways drove different aspects of disease in FHR/R mice with the C5aR pathway being responsible for macrovascular thrombosis and chronic inflammatory injury while the C5b-9 pathway caused renal TMA. Our data provide new understanding of the pathogenesis of complement-mediated TMA and macrovascular thrombosis in FHR/R mice and suggest that C5 blockade is more effective for the treatment of aHUS than selectively targeting the C5aR or C5b-9 pathway alone.

Murine systemic thrombophilia and hemolytic uremic syndrome from a factor H point mutation.

Complement plays a key role in host defense, but its dysregulation can cause autologous tissue injury. Complement activation is normally controlled by regulatory proteins, including factor H (FH) in plasma and membrane cofactor protein (MCP) on the cell surface. Mutations in FH and MCP are linked to atypical hemolytic uremic syndrome, a type of thrombotic microangiopathy (TMA) that causes renal failure. We describe here that disruption of FH function on the cell surface can also lead to disseminated complement-dependent macrovascular thrombosis. By gene targeting, we introduced a point mutation (W1206R) into murine FH that impaired its interaction with host cells but did not affect its plasma complement-regulating activity. Homozygous mutant mice carrying this mutation developed renal TMA as well as systemic thrombophilia involving large blood vessels in multiple organs, including liver, lung, spleen, and kidney. Approximately 30% of mutant mice displayed symptoms of stroke and ischemic retinopathy, and 48% died prematurely. Genetic deficiency of complement C3 and factor D prevented both the systemic thrombophilia and renal TMA phenotypes. These results demonstrate a causal relationship between complement dysregulation and systemic angiopathy and suggest that complement activation may contribute to various human thrombotic disorders involving both the micro- and macrovasculature.

Effect of Continued Administration of Low-dose Aspirin for Intraoperative Bleeding Control in Gastric Endoscopic Submucosal Dissection.

BACKGROUND: The use of antithrombotic agents for the prevention of cerebro-cardioembolic events has increased, and recent guidelines have recommended the continued administration of low-dose aspirin (LDA) during endoscopic procedures with a high risk of bleeding. However, the influence of LDA on intraoperative bleeding control status during Endoscopic submucosal dissection (ESD) remains unclear. METHODS: We examined 293 consecutive patients who underwent ESD for gastric cancers between January 2014 and February 2018. Patients administered with LDA (n = 52) were compared with those without antithrombotic therapy (n = 241; control) by propensity-score matching (PSM) concerning outcomes of ESD. RESULTS: PSM analysis yielded 50 matched pairs. Comparison showed similar values for frequency of intraoperative major bleeding: 1 (0-4) times (median [range]) in the LDA group and 0 (0-5) in the control group respectively (p = 0.710). Others (frequency of preventive coagulation, procedure time, decrease of hemoglobin levels, en bloc resection, complete resection) were the same with a few adverse events including perforation (0%), and thromboembolism (0%). Postoperative bleeding rate was 1.9% in LDA group. Multivariate analysis indicated that location U and circumference on the posterior wall were associated with for multiple major intraoperative bleeding. CONCLUSION: The study suggests that gastric ESD can be safely accomplished without cessation of LDA.

Blocking Properdin Prevents Complement-Mediated Hemolytic Uremic Syndrome and Systemic Thrombophilia.

Background Properdin (P) is a positive regulator of the alternative pathway of complement activation. Although P inhibition is expected and has been shown to ameliorate the alternative pathway of complement-mediated tissue injury in several disease models, it unexpectedly exacerbated renal injury in a murine model of C3 glomerulopathy. The role of P in atypical hemolytic uremic syndrome (aHUS) is uncertain.Methods We blocked P function by genetic deletion or mAb-mediated inhibition in mice carrying a factor H (FH) point mutation, W1206R (FHR/R), that causes aHUS and systemic thrombophilia with high mortality.Results P deficiency completely rescued FHR/R mice from premature death and prevented thrombocytopenia, hemolytic anemia, and renal disease. It also eliminated macrovessel thrombi that were prevalent in FHR/R mice. All mice that received a function-blocking anti-P mAb for 8 weeks survived the experimental period and appeared grossly healthy. Platelet counts and hemoglobin levels were significantly improved in FHR/R mice after 4 weeks of anti-P mAb treatment. One half of the FHR/R mice treated with an isotype control mAb but none of the anti-P mAb-treated mice developed stroke-related neurologic disease. Anti-P mAb-treated FHR/R mice showed largely normal renal histology, and residual liver thrombi were detected in only three of 15 treated mice.Conclusions These results contrast with the detrimental effect of P inhibition observed in a murine model of C3 glomerulopathy and suggest that P contributes critically to aHUS pathogenesis. Inhibition of P in aHUS may be of therapeutic benefit.

[DNA Extraction for Sensitive Detection of Shiga Toxin-Producing Escherichia coli in Food by Real-Time PCR Assays].

Alkali-heat DNA extraction, a rapid and economical method, was evaluated for use in the detection of Shiga toxin-producing Escherichia coli in food using real-time PCR assays. Alkali-heat DNA extracts led to highly sensitive detection (102-104 CFU/mL) of stx and O-antigen genes in beef liver, ground beef, sliced pork, cheese, lettuce, radish sprouts, tomato, and spinach, equivalent to the sensitivity obtained using a commercial DNA extraction kit that utilizes proteinase K lysis, and silica membrane purification. Although there were differences in DNA concentration and purity between DNA extraction methods, the sensitivity of real-time PCR assays was similar. These results indicate that alkali-heat DNA extraction is a viable method when testing food products with real-time PCR assays for the presence of stx and O-antigen genes.