Humanized CD19 CAR-T cells in relapsed/refractory B-ALL patients who relapsed after or failed murine CD19 CAR-T therapy.

BACKGROUND: For CD19-positive relapsed/refractory B-cell acute lymphoblastic leukemia (r/r B-ALL) after treatment with murine CD19 (mCD19) CAR-T, the reinfusion of mCD19 CAR-T cells may be ineffective due to anti-mouse single-chain variable fragment (scFv) antibody caused by mCD19 CAR. To overcome this immunogenicity, we applied humanized CD19 (hCD19) CAR-T cells to treat r/r B-ALL patients with prior mCD19 CAR-T therapy. METHODS: Nineteen pediatric and adult patients were included, 16 relapsed after and 3 were primarily resistant to mCD19 CAR-T. All patients presented with more than 5% blasts in bone marrow and/or extramedullary disease, and still showed CD19 antigen expression. Humanized CD19-CARs were lentiviral vectors carrying a second generation CAR with 4-1-BB co-stimulatory and CD3ζ signaling domains. Patient-derived cells were collected for producing CAR-T cells, the median dose of infused hCD19 CAR-T cells was 2.4 × 105/kg (range, 1.0-18.0 × 105/kg). RESULTS: hCD19 CAR-T resulted in a complete remission (CR) rate of 68% (13/19). Among 13 remission patients, 11 underwent allogeneic hematopoietic cell transplantation (allo-HCT) (3 were second HCT) and 10 remained in CR; the event-free survival rates at 12-18 months were 91% in 11 patients received following allo-HCT and 69% in all CR patients. Six cases had no response to hCD19 CAR-T, 3 died of disease progression; another 3 received salvage second transplantation, of them, 2 relapsed again (one died). Cytokine release syndrome (CRS) occurred in 95% (18/19) of patients, most CRS events were grade 1 and grade 2 (n = 17), there was only one grade 4 CRS. Two cases experienced grade 1 neurotoxicity. CONCLUSIONS: Humanized CD19 CAR-T cell therapy could be a treatment option for CD19-positive B-ALL patients who relapsed after or resisted prior murine CD19 CAR-T, hCD19 CAR-T followed by allo-HCT provided a longer remission in CR patients. Nevertheless, the prognosis of non-responders to hCD19 CAR-T remained dismal. TRIAL REGISTRATION: Chinese Clinical Trial Registry/WHO International Clinical Trial Registry ( ChiCTR1900024456 , URL: www.chictr.org.cn ); registered on July 12, 2019.

Combination of CD19 and CD22 CAR-T cell therapy in relapsed B-cell acute lymphoblastic leukemia after allogeneic transplantation.

The prognosis of relapsed acute lymphoblastic leukemia (ALL) after allogeneic transplantation is dismal when treated with conventional approaches. While single-target CD19 or CD22 chimeric antigen receptor (CAR) T-cell therapy has achieved high complete remission (CR) rates in refractory/relapsed B-ALL, it could not maintain a durable remission in most patients. To prolong relapse-free survival, we sequentially combined CD19 and CD22 CAR-T cells to treat post-transplant relapsed B-ALL patients with both CD19/CD22 antigen expression on lymphoblasts. Patient-derived donor cells were collected to produce CAR-T cells that were transfected by lentiviral vectors encoding second generation CARs composed of CD3ζ and 4-1BB. The second T-cell infusion was scheduled at least 1 month, and usually within 6 months after the first CAR-T treatment. Twenty-seven adult and pediatric patients, including 11 (41%) with extramedullary diseases (EMD), received the first CD19 CAR-T and 23 (85%) achieved CR. Subsequently, 21 out of 27 patients received the second CD22 CAR-T and were followed-up for a median of 19.7 (range, 5.6-27.3) months; 14 cases remained in CR, seven relapsed and two of them died from disease progression; Kaplan-Meier survival analysis showed overall survival and event-free survival rates of 88.5% and 67.5%, respectively, at both 12 months and 18 months. CAR-T associated graft-versus-host disease (GVHD) occurred in 23% of patients, with 8% new-onset acute GVHD and 15% persistent or worsened pre-existing cGVHD before CAR-T. This combination strategy of sequential CD19 and CD22 CAR-T therapy significantly improved the long-term survival in B-ALL patients who relapsed after transplantation.

Development of an efficient neutralization assay for Coxsackievirus A10.

Coxsackievirus A10 (CVA10) recently has become one of the major pathogens of hand, foot, and mouth disease (HFMD) in children worldwide, but no cure or vaccine against CVA10 is available yet. Serological evaluation of herd immunity to CVA10 will promote the development of vaccine. The traditional neutralization assay based on inhibition of cytopathic effect (Nt-CPE) is a common method for measuring neutralizing antibody titer against CVA10, which is time-consuming and labor-intensive. In this study, an efficient neutralization test based on a monoclonal antibody (mAb) 3D1 against CVA10, called Elispot-based neutralization test (Nt-Elispot), was developed. In the Nt-Elispot, the mAb 3D1 labeled with horseradish peroxidase (HRP) was used to detect the CVA10-infected RD cells at a 1:4000 dilution and the optimal infectious dose of CVA10 was set at 105 TCID50/well when combined with a fixed incubation time of 14 h. Compared with the Nt-CPE, the Nt-Elispot method effectively shortened the detection period and presented a good correlativity with it. Using the Nt-Elispot, a total of 123 sera from healthy children were tested for neutralizing antibody against CVA10, demonstrating that the overall seroprevalence was 49.3% (54/123) and the geometric mean titer (GMT) had been calculated as 574.2. Furthermore, 2 anti-CVA10 neutralizing mAbs were obtained by screening via the Nt-Elispot. Overall, the established Nt-Elispot could be used as an efficient and high-throughput method for evaluating immunity to CVA10 and screening the neutralizing antibodies.

Enhanced enantioselectivity of native α-cyclodextrins by the synergy of chiral ionic liquids in capillary electrophoresis.

In the cyclodextrins family, the native α-cyclodextrin has almost been abandoned in capillary electrophoresis chiral separation due to its much weaker enantioselectivity compared with ß-cyclodextrin and their derivatives. In this work, several amino acid chiral ionic liquids were selected to establish synergistic enantioseparation systems with native α-cyclodextrin. Enhanced enantioselectivities were observed in the chiral ionic liquids/α-cyclodextrin synergistic systems compared with single α-cyclodextrin system. A series of comparison experiments were performed to demonstrate the superiority of the synergistic systems. Primary parameters affecting the enantioseparation were systematically optimized, including the type and concentration of chiral ionic liquids, α-cyclodextrin concentration, buffer pH, and applied voltage. Best separations of the model enantiomers were obtained in a 20 mM Tris/H3 PO4 buffer at pH 2.5 containing 3% (m/v) α-cyclodextrin and 30 mM tetramethylammonium-l-arginine. The results show that the α-cyclodextrin is also worth our attention when selecting chiral selectors for capillary electrophoresis enantioseparation of specific racemic compound.

[Isolation and identification of an antimicrobial endophytic actinomycete from Macleaya cordata].

OBJECTIVE: An antimicrobial endophytic actinomycete strain was isolated from the root of Macleaya cordata, and its phylogenetic position was also investigated. METHODS: Endophytic actinomycetes were isolated from the root of Macleaya cordata growing in Dabie Mountain by tablet coating method. Antimicrobial activities of endophytic actinomycetes were determined by the agar block method,and then a potential strain was identified by morphology, biochemical characterization and molecular methods. RESULTS: A total of 15 strains of endophytic actinomycetes were isolated from the samples, six of them had antimicrobial activity to the test strains which accounts for 40% of all strains. Among the six strains,the strain BL7 was found to produce antibiotic substances which showed significant inhibitory effects on Staphylococcus aureus and Bacillus subtilis,with a formed inhibition zone of 29 mm and 20 mm respectively. Based on the morphology, biochemical characterization and 16S rDNA sequence analysis,the strain BL7 was preliminary identified as Streptomyces mobaraensis. CONCLUSION: Some members of Streptomyces genus have a very wide application prospect in medicine. This is the first report that Streptomyces mobaraensis strain was isolated from Macleaya cordata, and this study provides basis for the further exploration of the strain.

Investigating the virulence of coxsackievirus B6 strains and antiviral treatments in a neonatal murine model.

Coxsackievirus B6 (CVB6), a member of the human enterovirus family, is associated with severe diseases such as myocarditis in children. However, to date, only a limited number of CVB6 strains have been identified, and their characterization in animal models has been lacking. To address this gap, in this study, a neonatal murine model of CVB6 infection was established to compare the replication and virulence of three infectious-clone-derived CVB6 strains in vivo. The results showed that following challenge with a lethal dose of CVB6 strains, the neonatal mice rapidly exhibited a series of clinical signs, such as weight loss, limb paralysis, and death. For the two high-virulence CVB6 strains, histological examination revealed myocyte necrosis in skeletal and cardiac muscle, and immunohistochemistry confirmed the expression of CVB6 viral protein in these tissues. Real-time PCR assay also revealed higher viral loads in the skeletal and cardiac muscle than in other tissues at different time points post infection. Furthermore, the protective effect of passive immunization with antisera and a neutralizing monoclonal antibody against CVB6 infection was evaluated in the neonatal mouse model. This study should provide insights into the pathogenesis of CVB6 and facilitate further research in the development of vaccines and antivirals against CVBs.

The carbon reduction channel through which financing methods affect total factor productivity: mediating effect tests from 23 major carbon-emitting countries.

The development of modern finance has played a catalytic role in the economic transition to renewable and clean energy, which in turn has an impact on total factor productivity (TFP). However, existing studies have not together addressed financing methods, carbon emissions, and TFP. We analyse how different financing methods affect TFP through the carbon reduction channel. Using data from 1995 to 2019 for 23 major carbon emitters, we adopt a mediation effect model with Stata 17.0. We draw three conclusions. First, financing methods have a differentiated impact on TFP. For every unit increase in debt financing relative to equity financing, TFP decreases by 0.058 units (overall level) or 0.056 units (welfare level). Second, financing has a mediating effect on TFP through carbon emissions. Debt financing reduces TFP through the carbon emission reduction mechanism. The greater the scale of debt financing relative to equity financing, the greater the negative impact on TFP through the carbon emission reduction mechanism, while equity financing plays a positive role on TFP through the carbon emission reduction mechanism. Third, a heterogeneity test demonstrates that the mediating effect is most significant in developed countries and weakest in developing countries. The difference-in-difference framework based on the Equator Principles demonstrates that the marginal contribution to TFP of debt financing aimed at carbon reduction is 0.02 (overall level) and 0.012 (welfare level). From the perspective of financing methods, this study provides enlightenment for promoting carbon emission reduction and improving TFP. First, countries should strengthen the development of the green debt financing market, strengthen the disclosure of information on environmental benefits, and reverse the negative effect of debt financing. Second, they should develop the equity market to activate the role of carbon reduction channel, promote the Equator Principles in the banking industry, and encourage more banks to pay attention to environmental risks. All these financial measures can raise TFP.

CAR-T therapy as a consolidation in remission B-ALL patients with poor prognosis.

BACKGROUND: To date, almost all studies regarding chimeric antigen receptor (CAR)-T cell therapy for B-cell acute lymphoblastic leukemia (B-ALL) were performed in refractory/relapsed (r/r) or minimal residual disease-positive patients. CAR-T therapy in remission patients has not been reported. AIM: To observe the treatment outcome of CAR-T cells for remission B-ALL patients with poor prognosis. METHODS AND RESULTS: CAR-T treatment was applied to two B-ALL patients in remission status who had poor prognostic factors and refused transplantation, and one case was unable to accept standard chemotherapy owing to multiple complications. The procedure of CAR-T therapy in these two remission patients was the same as that in r/r B-ALL patients. Lentiviral vectors encoding second generation CARs composed of CD3ζ and 4-1BB were used to produce CAR-T cells. Lymphodepleting agents fludarabine and cyclophosphamide were administered prior to cell infusion. Grade I cytokine release syndrome occurred after each T-cell infusion and there was no neurotoxicity. CAR-T treatment followed by non-intensive maintenance chemotherapy and targeted drugs allowed both patients to obtain a long-term event-free survival of more than three and a half years without transplantation. CONCLUSIONS: CAR-T therapy could be used in high-risk B-ALL patients as a consolidation to avoid transplantation, the combination of CAR-T and following maintenance therapy may be better than CAR-T alone for durable remission.

Development of a rapid neutralization assay for the detection of neutralizing antibodies against coxsackievirus B1.

Coxsackievirus B1 (CVB1) is a major pathogen that causes viral myocarditis and aseptic meningitis and is implicated as a cause of type 1 diabetes mellitus. The rapid detection of neutralizing antibodies can help in the prevention and diagnosis of viral infection. The traditional cytopathic effect (CPE)-based neutralization assay (Nt-CPE) is time-consuming and labor-intensive. In this study, an efficient neutralization test based on an enzyme-linked immunospot assay and a monoclonal antibody 2E6 against CVB1 (Nt-Elispot) was developed. In this optimal Nt-Elispot, a multiplicity of infection (MOI) of 1 per well was set as the infection dose, and an incubation time of 18 hours was selected as the checkpoint. Compared with Nt-CPE, Nt-Elispot significantly shortened the detection period and displayed a good correlation with it. This established CVB1 Nt-Elispot could be applied to efficiently screen neutralizing antibodies and evaluate the level of NAb against CVB1 in large cohorts.

Development of a rapid neutralization testing system for Rhinovirus C15 based on the enzyme-linked immunospot assay.

Human Rhinoviruses (RVs) are dominant pathogens causing a wide range of respiratory tract diseases, posing a huge threat to public health worldwide. Viruses belonging to the RV-C species are more likely to cause severe illnesses and are strongly associated with asthma onset or exacerbations than RV-A or RV-B. Rapid and sensitive detection of neutralizing antibodies (NAbs) against RV-C can promote the development of vaccines and antiviral drugs and help in the diagnosis of viral infection. In this study, a rapid neutralization testing system for RV-C15, based on an enzyme-linked immunospot assay (Nt-ELISPOT) was developed. A monoclonal antibody (MAb), named 9F9, with high binding efficacy for RV-C15 conjugated to horseradish peroxidase (HRP), was used to detect RV-C15-infected cells at a concentration of 2 µg/ml. The optimal infectious dose of RV-C15 was set at 1 × 104 TCID50/well and the cells were fixed with 0.5% formaldehyde diluted in PBS after incubation for 20 h. Compared with the traditional cytopathic effect (CPE)-based neutralization assay (Nt-CPE), Nt-ELISPOT significantly shortened the detection period and showed good consistency with the detection of neutralizing titers of both sera and NAbs. Using Nt-ELISPOT, three anti-RV-C15 NAbs were obtained with IC50 values of 0.16, 0.27, and 11.8 µg/ml, respectively. Moreover, 64 human serum samples collected from a wide range of age groups were tested for NAb against RV-C15 by Nt-ELISPOT. The total seroprevalence was 48.4% (31/64) and the positive rate was lowest in the group under 6 years old. Thus, the Nt-ELISPOT established in this study can be used as a high-throughput and rapid neutralization assay for the screening of NAbs and for seroepidemiological investigation against RV-C15.

Structural basis for the synergistic neutralization of coxsackievirus B1 by a triple-antibody cocktail.

Coxsackievirus B1 (CVB1) is an emerging pathogen associated with severe neonatal diseases including aseptic meningitis, myocarditis, and pancreatitis and also with the development of type 1 diabetes. We characterize the binding and therapeutic efficacies of three CVB1-specific neutralizing antibodies (nAbs) identified for their ability to inhibit host receptor engagement. High-resolution cryo-EM structures showed that these antibodies recognize different epitopes but with an overlapping region in the capsid VP2 protein and specifically the highly variable EF loop. Moreover, they perturb capsid-receptor interactions by binding various viral particle forms. Antibody combinations achieve synergetic neutralization via a stepwise capsid transition and virion disruption, indicating dynamic changes in the virion in response to multiple nAbs targeting the receptor-binding site. Furthermore, this three-antibody cocktail protects against lethal challenge in neonatal mice and limits pancreatitis and viral replication in a non-obese diabetic mouse model. These results illustrate the utility of nAbs for rational design of therapeutics against picornaviruses such as CVB.

Reduced Intensity Conditioning Followed by Allogeneic Hematopoietic Stem Cell Transplantation Is a Good Choice for Acute Myeloid Leukemia and Myelodysplastic Syndrome: A Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: Reduced intensity conditioning (RIC) before allogeneic hematopoietic stem cell transplantation (allo-HSCT) has been reported to have the same overall survival (OS) as myeloablative conditioning (MAC) for patients with acute myeloid leukemia (AML) in complete remission (CR) and myelodysplastic syndrome (MDS). However, results from different studies are conflicting. Therefore, we conducted a systematic review and meta-analysis guided by PRISMA 2009 to confirm the efficacy and safety of RIC vs. MAC for AML in CR and MDS. METHODS: We search PubMed, Web of Science, Embase, Cochrane central, clinical trial registries and related websites, major conference proceedings, and field-related journals from January 1, 1980, to July 1, 2020, for studies comparing RIC with MAC before the first allo-HSCT in patients with AML in CR or MDS. Only randomized controlled trials (RCTs) were included. OS was the primary endpoint and generic inverse variance method was used to combine hazard ratio (HR) and 95% CI. RESULTS: We retrieved 7,770 records. Six RCTs with 1,413 participants (711 in RIC, 702 in MAC) were included. RIC had the same OS (HR = 0.95, 95% CI 0.64-1.4, p = 0.80) and cumulative incidence of relapse as MAC (HR = 1.18, 95% CI 0.88-1.59, p = 0.28). Furthermore, RIC significantly reduced non-relapse mortality more than total body irradiation/busulfan-based MAC (HR = 0.53, 95% CI 0.36-0.80, p = 0.002) and had similar long-term OS and graft failure as MAC. CONCLUSION: RIC conditioning regimens are recommended as an adequate option of preparative treatment before allo-HSCT for patients with AML in CR or MDS. SYSTEMATIC REVIEW REGISTRATION: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=185436.

Government Epidemic Prevention and Economic Growth Path Under Public Health Emergency: Theoretical Model and Simulation Analysis.

This paper constructs a partial equilibrium model under public health emergency shocks based on economic growth theory, and investigates the relationship between government intervention and virus transmission and economic growth path. We found that both close contacts tracing measures and isolation measures are beneficial to human capital stock and economic output per capita, and the effect of close contact tracing measures is better than that of isolation measures. For infectious diseases of different intensities, economic growth pathways differed across interventions. For low contagious public health emergencies, the focus should be on the coordination of isolation and tracing measures. For highly contagious public health emergencies, strict isolation, and tracing measures have limited effect in repairing the negative economic impact of the outbreak. The theoretical model provides a basic paradigm for the future researches to study economic growth under health emergencies, with good scalability and robustness.

The COVID-19 Pandemic and Economic Growth: Theory and Simulation.

The outbreak of COVID-19 pandemic has caused profound consequences on world economy. In order to explore the long-term impact of the pandemic on economic growth and the effects of different policy responses, this paper combines economic theory with epidemiological model to construct an interdisciplinary model, in which labor supply is dynamically constrained by pandemic conditions. Analysis of model equilibrium suggests that outbreaks of infectious disease reduce labor supply and negatively affect economic output. The accumulation of health capital can suppress the spread of disease and improve the recovery rate of infected individuals, which will alleviate the labor supply constraint caused by the pandemic and lead to an increase in output and consumption. The model is then calibrated to Chinese economy. The simulation results imply that government's public health policy can enhance the role of health capital in promoting economic growth. But the marginal effect of certain policies is diminishing. Therefore, the government needs to balance pandemic prevention and control costs and marginal benefits when formulating public health policies. When the pandemic is under control, the resumption of production is feasible and the economic stimulus package could lead to economic recovery.

Modeling the Global Dynamic Contagion of COVID-19.

The COVID-19 infections have profoundly and negatively impacted the whole world. Hence, we have modeled the dynamic spread of global COVID-19 infections with the connectedness approach based on the TVP-VAR model, using the data of confirmed COVID-19 cases during the period of March 23rd, 2020 to September 10th, 2021 in 18 countries. The results imply that, (i) the United States, the United Kingdom and Indonesia are global epidemic centers, among which the United States has the highest degree of the contagion of the COVID-19 infections, which is stable. South Korea, France and Italy are the main receiver of the contagion of the COVID-19 infections, and South Korea has been the most severely affected by the overseas epidemic; (ii) there is a negative correlation between the timeliness, effectiveness and mandatory nature of government policies and the risk of the associated countries COVID-19 epidemic affecting, as well as the magnitude of the net contagion of domestic COVID-19; (iii) the severity of domestic COVID-19 epidemics in the United States and Canada, Canada and Mexico, Indonesia and Canada is almost equivalent, especially for the United States, Canada and Mexico, whose domestic epidemics are with the same tendency; (iv) the COVID-19 epidemic has spread though not only the central divergence manner and chain mode of transmission, but also the way of feedback loop. Thus, more efforts should be made by the governments to enhance the pertinence and compulsion of their epidemic prevention policies and establish a systematic and efficient risk assessment mechanism for public health emergencies.

Development of A Neonatal Mouse Model for Coxsackievirus B1 Antiviral Evaluation.

Coxsackievirus B1 (CVB1) is a leading causative agent of severe infectious diseases in humans and has been reported to be associated with outbreaks of aseptic meningitis, myocarditis, and the development of chronic diseases such as type 1 diabetes mellitus (T1DM). There is no approved vaccine or effective antiviral therapy to treat CBV1 infection. And animal models to assess the effects of antiviral agents and vaccine remain limited. In this study, we established a neonatal mouse model of CVB1 using a clinically isolated strain to characterize the pathological manifestations of virus infection and to promote the development of vaccines and antiviral drugs against CVB1. One-day-old BALB/c mice were susceptible to CVB1 infection by intraperitoneal injection. Mice challenged with CVB1 at a low dose [10 median tissue culture infective dose (TCID50)] exhibited a series of clinical symptoms, such as inactivity, emaciation, limb weakness, hair thinning, hunching and even death. Pathological examination and tissue viral load analysis showed that positive signals of CVB1 were detected in the heart, spinal cord, limb muscle and kidney without pathological damage. Particularly, CVB1 had a strong tropism towards the pancreas, causing severe cellular necrosis with inflammatory infiltration, and was spread by viraemia. Notably, the monoclonal antibody (mAb) 6H5 and antisera elicited from CVB1-vaccinated mice effectively protected the mice from CVB1 infection in the mouse model. In summary, the established neonatal mouse model is an effective tool for evaluating the efficacy of CVB1 antiviral reagents and vaccines.

Cryo-EM structures reveal the molecular basis of receptor-initiated coxsackievirus uncoating.

Enterovirus uncoating receptors bind at the surface depression ("canyon") that encircles each capsid vertex causing the release of a host-derived lipid called "pocket factor" that is buried in a hydrophobic pocket formed by the major viral capsid protein, VP1. Coxsackievirus and adenovirus receptor (CAR) is a universal uncoating receptor of group B coxsackieviruses (CVB). Here, we present five high-resolution cryoEM structures of CVB representing different stages of virus infection. Structural comparisons show that the CAR penetrates deeper into the canyon than other uncoating receptors, leading to a cascade of events: collapse of the VP1 hydrophobic pocket, high-efficiency release of the pocket factor and viral uncoating and genome release under neutral pH, as compared with low pH. Furthermore, we identified a potent therapeutic antibody that can neutralize viral infection by interfering with virion-CAR interactions, destabilizing the capsid and inducing virion disruption. Together, these results define the structural basis of CVB cell entry and antibody neutralization.

Childhood familial environment and adulthood depression: evidence from a Chinese population-based study.

BACKGROUND: Mental disorders have become an important public health issue and evidence is lacking on the impact of childhood experience on adulthood mental health in regions of low and middle income. Using national representative data from the China Health and Retirement Longitudinal Study, we aimed to explore the impact of childhood familial environment on adulthood depression. METHODS: A total of 19 485 subjects were interviewed. The survey collected information on demographic variables, variables of childhood familial environment and potential pathway variables, including childhood health status, adulthood physical health status, adulthood social support and adulthood socio-economic status (SES). Depressive symptoms were measured by the 10-item version of the Center for Epidemiological Studies Depression Scale. RESULTS: Parents' physical and mental health during the subjects' childhood were significantly associated with adulthood mental health. Mothers' smoking, unfair treatment and low family SES were associated with higher depressive symptoms in adulthood. Childhood physical and mental health status, adulthood physical health and adulthood SES might be important mediators in the pathways of childhood familial environment affecting adulthood depressive symptoms. CONCLUSIONS: This study is the first to explore the relationship of childhood familial environment and adulthood depression in China. The results indicate that parents' physical and mental health, health behaviour and treatment equity among children a important predictors for adult depression.

A new approach to removing and recovering phosphorus from livestock wastewater using dolomite.

Recovering phosphorus from livestock wastewater could partly mitigate the global phosphorus resource crisis. Crystallization is a promising method for removing phosphorus from wastewater, but the costs of calcium- and magnesium-containing reagents are increasing. Cheap, available, efficient materials are required to replace conventional calcium and magnesium reagents. Here, we describe a new approach to removing and recovering phosphorus from livestock wastewater of a large pig farm, containing a high phosphorus concentration. The effects of the pH, stirring speed, stirring time, and extract dose (containing calcium and magnesium) on phosphorus removal from livestock wastewater were investigated. The product was characterized by X-ray diffractometry, Fourier-transform infrared spectroscopy, and scanning electron microscopy. Under optimized conditions (pH 9.0, stirring speed 200 r/m, stirring time 600 s, Ca 207.62 mg/L, Mg 122.86 mg/L), 92% of the phosphorus was removed from livestock wastewater. The product was mainly the hydroxyapatite (Ca5(PO4)3OH) precursor amorphous calcium phosphate but also contained 1.65% (by mass) magnesium ammonium phosphate (MgNH4PO4·6H2O) crystals. The cost of dolomite to treat 1 m3 of high-phosphorus wastewater was 0.20 yuan (45.9%, 25.9%, and 75.9% lower than for pure MgCl2, MgSO4, and CaCl2, respectively) in 2019. Using dolomite to provide calcium and magnesium effectively decreases the crystallization process cost and should encourage the use of crystallization to remove phosphorus from wastewater.

Identification of Antibodies with Non-overlapping Neutralization Sites that Target Coxsackievirus A16.

Hand, foot, and mouth disease is a common childhood illness primarily caused by coxsackievirus A16 (CVA16), for which there are no current vaccines or treatments. We identify three CVA16-specific neutralizing monoclonal antibodies (nAbs) with therapeutic potential: 18A7, 14B10, and NA9D7. We present atomic structures of these nAbs bound to all three viral particle forms-the mature virion, A-particle, and empty particle-and show that each Fab can simultaneously occupy the mature virion. Additionally, 14B10 or NA9D7 provide 100% protection against lethal CVA16 infection in a neonatal mouse model. 18A7 binds to a non-conserved epitope present in all three particles, whereas 14B10 and NA9D7 recognize broad protective epitopes but only bind the mature virion. NA9D7 targets an immunodominant site, which may overlap the receptor-binding site. These findings indicate that CVA16 vaccines should be based on mature virions and that these antibodies could be used to discriminate optimal virion-based immunogens.