Multiplexed activation of endogenous genes by CRISPRa elicits potent antitumor immunity.

Immunotherapy has transformed cancer treatment. However, current immunotherapy modalities face various limitations. In the present study, we developed multiplexed activation of endogenous genes as an immunotherapy (MAEGI), a new form of immunotherapy that elicits antitumor immunity through multiplexed activation of endogenous genes in tumors. We leveraged CRISPR activation (CRISPRa) to directly augment the in situ expression of endogenous genes, and thereby the presentation of tumor antigens, leading to dramatic antitumor immune responses. Deploying this as a cell-based vaccination strategy showed efficacy in both prophylactic and therapeutic settings. Intratumoral adeno-associated virus delivery of CRISPRa libraries elicited strong antitumor immunity across multiple cancer types. Precision targeting of mutated gene sets eradicated a large fraction of established tumors at both local and distant sites. This treatment modality led to alterations in the tumor microenvironment, marked by enhanced T cell infiltration and antitumor immune signatures. Multiplexed endogenous gene activation is a versatile and highly scalable strategy to elicit potent immune responses against cancer, distinct from all existing cancer therapies.

Electrochemical Failure Mechanism of δ-MnO2 in Zinc Ion Batteries Induced by Irreversible Layered to Spinel Phase Transition.

Phase transitions of Mn-based cathode materials associated with the charge and discharge process play a crucial role on the rate capability and cycle life of zinc ion batteries. Herein, a microscopic electrochemical failure mechanism of Zn-MnO2 batteries during the phase transitions from δ-MnO2 to λ-ZnMn2O4 is presented via systematic first-principle investigation. The initial insertion of Zn2+ intensifies the rearrangement of Mn. This is completed by the electrostatic repulsion and co-migration between guest and host ions, leading to the formation of λ-ZnMn2O4. The Mn relocation barrier for the λ-ZnMn2O4 formation path with 1.09 eV is significantly lower than the δ-MnO2 re-formation path with 2.14 eV, indicating the irreversibility of the layered-to-spinel transition. Together with the phase transition, the rearrangement of Mn elevates the Zn2+ migration barrier from 0.31 to 2.28 eV, resulting in poor rate performance. With the increase of charge-discharge cycles, irreversible and inactive λ-ZnMn2O4 products accumulate on the electrode, causing continuous capacity decay of the Zn-MnO2 battery.

An amino-rich polymer-coated magnetic nanomaterial for ultra-rapid separation of phosphorylated peptides in the serum of Parkinson's disease patients.

The elucidation of disease pathogenesis can be achieved by analyzing the low-abundance phosphopeptides in organisms. Herein, we developed a novel and easy-to-prepare polymer-coated nanomaterial. By improving the hydrophilicity and spatial conformation of the material, we effectively enhanced the adsorption of phosphopeptides and demonstrated excellent enrichment properties. The material was able to successfully enrich the phosphopeptides in only 1 min. Meanwhile, the material has high selectivity (1:2000), good loading capacity (100 µg/mg), excellent sensitivity (0.5 fmol), and great acid and alkali resistance. In addition, the material was applied to real samples, and 70 phosphopeptides were enriched from the serum of Parkinson's disease (PD) patients and 67 phosphopeptides were enriched from the serum of normal controls. Sequences Logo showed that PD is probably associated with threonine, glutamate, serine, and glutamine. Finally, gene ontology (GO) analysis was performed on phosphopeptides enriched in PD patients' serum. The results showed that PD patients expressed abnormal expression of the cholesterol metabolic process and cell-matrix adhesion in the biological process (BP), endoplasmic reticulum and lipoprotein in the cellular component (CC), and heparin-binding, lipid-binding, and receptor-binding in the molecular function (MF) as compared with normal individuals. All the experiments indicate that the nanomaterials have great potential in proteomics studies.

Biallelic mutations in ARMC12 cause asthenozoospermia and multiple midpiece defects in humans and mice.

BACKGROUND: Asthenozoospermia is a major factor contributing to male infertility. The mitochondrial sheath (MS), an important organelle in the midpiece of spermatozoa, is crucial to sperm motility. ARMC12 is a mitochondrial peripheral membrane protein. Deletion of Armc12 impairs the arrangement of MS and causes infertility in mice. However, the role of ARMC12 in human asthenozoospermia remains unknown. OBJECTIVE: To study the genetic defects in patients with asthenozoospermia. METHODS: A total of 125 patients with asthenozoospermia and 120 men with proven fertility were recruited. Whole-exome sequencing and Sanger sequencing were performed for genetic analysis. Papanicolaou staining, HE staining, immunofluorescent staining, transmission electron microscopy and field emission scanning electron microscopy were employed to observe the morphological and structural defects of the spermatozoa and testes. Armc12-knockout mice were generated using the CRISPR-Cas9 system. Intracytoplasmic sperm injection was used to treat the patients. RESULTS: Biallelic ARMC12 mutations were identified in three patients, including homozygous mutations in two siblings from a consanguineous family and compound heterozygous mutations in one sporadic patient. ARMC12 is mainly expressed in the midpiece of elongated and late spermatids in the human testis. The patients' spermatozoa displayed multiple midpiece defects, including absent MS and central pair, scattered or forked axoneme and incomplete plasma membrane. Spermatozoa from Armc12-/- mice showed parallel defects in the midpiece. Moreover, two patients were treated with intracytoplasmic sperm injection and achieved good outcomes. CONCLUSION: Our findings prove for the first time that defects in ARMC12 cause asthenozoospermia and multiple midpiece defects in humans.

Community exposure to gun homicide and adolescents' educational aspirations.

INTRODUCTION: Witnessing violence and violent victimization have detrimental effects on adolescents' emotional functioning and ability to envision and plan for their futures. However, research is limited on the impact of violence that occurs in adolescents' communities-whether or not it was witnessed or experienced firsthand. This paper investigated the associations between community exposure to gun homicide and adolescents' high school and college graduation aspirations. METHODS: We analyzed data from the Future of Families and Child Wellbeing Study (N = 3031), a cohort study of children born 1998-2000 in 20 large US cities, merged with incident-level data on deadly gun violence from the Gun Violence Archive (2014-2017). Outcomes were reported by adolescents (girls and boys) during wave 6 (2014-2017) of the study, conducted when the children were 15 years of age. We employed ordinary least squares regression, ordered logistic regression, and multilevel stratification to examine the average and heterogeneous impacts of community exposure to gun homicide on adolescents' educational aspirations. RESULTS: Community exposure to gun homicide was associated with reduced high school graduation aspirations, particularly among adolescents with the lowest risk of exposure to gun homicide. Gun homicide exposure was also associated with increased college graduation aspirations; this association was concentrated among adolescents with moderate-high risk of exposure. CONCLUSIONS: Given the importance of education for job opportunities and the better health that accompanies education and occupational attainment, preventing early exposure to gun violence and providing institutional supports to help adolescents facing adversity realize their goals is essential to their long-term health and success.

Defect-Engineered VS2 Electrocatalysts for Lithium-Sulfur Batteries.

Defective two-dimensional transition metal dichalcogenides can be effective electrocatalysts for Li-S batteries, but the relationship between defect types and battery performance is unclear. In this work, we designed S vacancy-type SV-VS2 and V self-intercalated-type VI-VS2 and measured their catalytic activities in Li-S batteries. Compared with self-intercalating V atoms, S vacancies accelerated Li+ diffusion and SV-VS2 as a Li+ "reservoir" promoted the sulfur conversion kinetics significantly. In addition, the presence of sulfur vacancies promoted the lithiation behavior of SV-VS2 during discharge, leading to an enhancement of the catalytic ability of SV-VS2. However, this lithiation phenomenon weakened the catalytic activity of VI-VS2. Overall, SV-VS2 had better adsorption and catalytic activity. Li-S batteries with SV-VS2-coated separators delivered high rate performance and excellent cycling stability, with a capacity decay rate of 0.043% over 880 cycles at 1.0 C. This work provides an effective strategy for designing efficient Li-S battery electrocatalysts using defect engineering.

Uniform Zinc Deposition Regulated by a Nitrogen-Doped MXene Artificial Solid Electrolyte Interlayer.

The dendrite growth and side reactions of zinc metal anode in mildly acidic electrolytes seriously hinder the practical application of aqueous zinc-ion battery. To address these issues, an artificial protective layer of nitrogen-doped MXene (NMX) is used to protect the zinc anode. The NMX protective layer has high conductivity and uniformly distributed zincophilic sites, which can not only homogenize the local electric field on the electrode interface but also accelerate the kinetics for Zn deposition. As a result, the NMX protective layer induces uniform zinc deposition and reduces the overpotential of the electrode. Encouragingly, this NMX-protected Zn anode can cycle stably for 1900 h at 1 mA cm-2 and 1 mAh cm-2 . In asymmetric cells, it achieves high cycle reversibility with an average Coulomb efficiency of 99.79% for 4800 cycles at 5 mA cm-2 .

A novel hydrophilic polymer-coated magnetic nanomaterial based on the HILIC strategy for fast separation of glycopeptides and glycosylated exosomes.

Novel hydrophilic poly(N, N-methylenebisacrylamide/1,2-epoxy-5-hexene) coated magnetic nanospheres functionalized with 2-aminopurine (denoted as Fe3O4@poly(MBA/EH)@2AP) for enriching glycopeptides and glycosylated exosomes were successfully obtained using a simple and green method on the basis of the HILIC (hydrophilic interaction liquid chromatography) enrichment strategy. The high density of polar groups endows the material with amazing hydrophilicity, enabling the nanomaterial to successfully capture glycopeptides and glycosylated exosomes within 1 min. Meanwhile, the materials demonstrated great sensitivity (0.01 fmol/µL), good loading capability (125 µg/mg), high selectivity (BSA:HRP = 1000:1), and repeatability (more than 10 times). Besides, the material was applied in the analysis of bio-samples, a total of 290 glycosylated peptides and 184 glycosylation sites mapping to 185 glycoproteins were identified in the serum of uremic patients. Besides, 42 glycopeptides were enriched from the saliva of healthy people. At the same time, it was verified by TEM and western blot that the complete glycosylated exosomes were successfully captured from the serum of the uremic patients. All experiments have demonstrated that Fe3O4@poly(MBA/EH)@2AP has a promising future in practical applications.

Beyond orchids and dandelions: Susceptibility to environmental influences is not bimodal.

This study focused on generality versus specificity of susceptibility of effects of eight family and child-care exposures measured between 3 and 54 months of age (e.g., sensitive parenting, child-care quality) on five child development outcomes assessed at age 4.5 years (e.g. behavior problems, preacademic skill), using data from The National Institute of Child Health and Human Development (NICHD) Study of Early Child Care and Youth Development (n = 1,364, boys = 705; White = 1,097, Black = 176, other = 91), while applying a novel influence-statistics method. Results indicated that susceptibility across the environment-predictor:child-outcome associations is normally rather than bimodally (i.e., orchid-dandelion) distributed. Analysis of susceptibility documents both domain generality and specificity of developmental plasticity, with effect sizes proving small in the former case. As predicted, children who as infants had difficult temperaments or who scored higher on a polygenic-plasticity score (serotonin-transporter-linked promoter region [5-HTTLPR], dopamine receptor D4 [DRD4], brain-derived neurotrophic factor [BDNF]) proved somewhat more susceptible to some of the environmental effects investigated. Results lead to the recommendation that two-types-of-individuals vis-a-vis susceptibility to environmental influences be questioned and general-trait conceptions of susceptibility be further investigated.

Fabrication of a polymer brush-functionalized porphyrin-based covalent organic framework for enrichment of N-glycopeptides.

A surface-initiated atom transfer radical polymerization method combining click chemistry was employed to prepare a novel porphyrin-based covalent organic framework composite grafted with polymer brushes (TAPBB@GMA@AMA@Cys) for the specific enrichment of N-glycopeptides. The material successfully realized the high efficiency enrichment of N-glycopeptides with good selectivity (1:1000), low detection limit (0.2 fmol/µL), and high loading capacity (133.3 mg·g-1). The TAPBB@GMA@AMA@Cys was successfully applied to actual sample analysis; 235 N-glycopeptides related to 125 glycoproteins and 210 N-glycopeptides related to 121 glycoproteins were recognized from the serum of normal individuals and Alzheimer's disease patients, respectively. Gene ontology studies of molecular functions, cellular components, and biological processes have revealed that identified glycoproteins are strongly associated with neurodegenerative diseases involving innate immune responses, basement membranes, calcium binding, and receptor binding. The above results confirm the surprising potential of materials in glycoproteomics research and practical sample applications.

Ti4+ functionalized ß-cyclodextrin covalent organic framework as a new immobilized metal ion affinity chromatography platform for selective capture of phosphorylated peptides and exosomes.

A Ti4+ functionalized ß-cyclodextrin covalent organic framework nanoparticle (named as ß-CD-COF@Ti4+) was synthesized using a one-pot method successfully realizing the enrichment of phosphorylated peptides and exosomes based on the immobilized metal ion affinity chromatography strategy. The functionalized ß-CD-COF@Ti4+ exhibited superior performance on the enrichment of phosphopeptides, including high selectivity (1:1000), low detection limit (0.5 fmol), and loading capacity for phosphopeptides (100 mg·g-1). After treatment with ß-CD-COF@Ti4+, 9 phosphopeptides from defatted milk, 29 phosphopeptides related to 23 phosphoproteins from normal group serum, and 24 phosphopeptides related to 22 phosphoproteins from the serum of uremia patients were captured. Through the analysis of Gene Ontology, the captured phosphoprotein is closely related to kidney disease, including lipoprotein metabolism, very-low-density lipoprotein particle, high-density lipoprotein particle, and lipid binding activity process. Furthermore, western blot verification showed that this nanoparticle could successfully capture exosomes from human serum. This study demonstrates great prospects for the enrichment of phosphopeptides and exosomes from actual bio-samples.

Binding properties of odorant-binding protein 4 from bean bug Riptortus pedestris to soybean volatiles.

The bean bug Riptortus pedestris is a notorious insect pest that can damage various crops, especially soybean, in East Asia. In insects, the olfactory system plays a crucial role in host finding and feeding behaviour in which the odorant-binding proteins (OBPs) are believed to be involved in initial step in this system. In this study, we produced the R. pedestris adult antennae-expressed RpedOBP4 protein using a recombinant expression system in E. coli. Fluorescence competitive binding confirmed that RpedOBP4 has binding affinities to 7 of 20 soybean volatiles (ligands), and that a neutral condition is the best environment for it. The binding property of RpedOBP4 to these ligands was further revealed by integrating data from molecular docking, site-directed mutagenesis and ligand binding assays. This demonstrated that five amino acid residues (I30, L33, Y47, I57 and Y121) are involved in the binding process of RpedOBP4 to corresponding ligands. These findings will not only help us to more thoroughly explore the olfactory mechanism of R. pedestris during feeding on soybean, but also lead to the identification of key candidate targets for developing environmental and efficient behaviour inhibitors to prevent population expansion of R. pedestris in the future.

Heterogeneous effects of spatially proximate firearm homicide exposure on anxiety and depression symptoms among U.S. youth.

The burden of firearm homicide in the United States is not evenly distributed across the population; rather, it disproportionately affects youth in disadvantaged and marginalized communities. Research is limited relevant to the impacts of exposure to firearm violence that occurs near where youth live or attend school - spatially proximate firearm violence - on youths' mental health and whether those impacts vary by characteristics that shape youths' risk for experiencing that exposure in the first place. Using a dataset linking the Fragile Families and Child Wellbeing Study with the Gun Violence Archive (N = 3086), we employed propensity score matching and multilevel stratification to examine average and heterogeneous associations between spatially proximate firearm homicide exposure and anxiety and depression among all youth and then separately for boys and girls. We found a statistically significant average association between firearm homicide exposure and symptoms of depression among youth. Furthermore, heterogeneous effects analyses yielded evidence that the average association is driven by youth, and particularly boys, who are the most disadvantaged and have the highest risk of firearm homicide exposure. The results of this study suggest that the accumulation of stressors associated with structural disadvantage and neighborhood disorder, coupled with exposure to spatially proximate and deadly firearm violence, may make boys and young men, particularly Black boys and young men, uniquely vulnerable to the mental health impacts of such exposure. Ancillary analyses of potential effect moderators suggest possible future areas of investigation.

Inequities in Community Exposure to Deadly Gun Violence by Race/Ethnicity, Poverty, and Neighborhood Disadvantage among Youth in Large US Cities.

Understanding the burden of gun violence among youth is a public health imperative. While most estimates are based on direct and witnessed victimization, living nearby gun violence incidents may be consequential too. Yet detailed information about these broader experiences of violence is lacking. We use data on a population-based cohort of youth merged with incident-level data on deadly gun violence to assess the prevalence and intensity of community exposure to gun homicides across cross-classified categories of exposure distance and recency, overall and by race/ethnicity, household poverty, and neighborhood disadvantage. In total, 2-18% of youth resided within 600 m of a gun homicide occurring in the past 14-365 days. These percentages were 3-25% for incidents within 800 m and 5-37% for those within a 1300-m radius. Black and Latinx youth were 3-7 times more likely, depending on the exposure radius, to experience a past-year gun homicide than white youth and on average experienced incidents more recently and closer to home. Household poverty contributed to exposure inequities, but disproportionate residence in disadvantaged neighborhoods was especially consequential: for all racial/ethnic groups, the difference in the probability of exposure between youth in low vs high poverty households was approximately 5-10 percentage points, while the difference between youth residing in low vs high disadvantage neighborhoods was approximately 50 percentage points. Given well-documented consequences of gun violence exposure on health, these more comprehensive estimates underscore the importance of supportive strategies not only for individual victims but entire communities in the aftermath of gun violence.

A Ti/Nb-functionalized COF material based on IMAC strategy for efficient separation of phosphopeptides and phosphorylated exosomes.

In this work, on the basis of an immobilized metal ion affinity chromatography enrichment strategy, a new kind of covalent organic framework (COF) material for enrichment of phosphorylated peptides and exosomes was successfully prepared in a facile method, and Ti4+ and Nb5+ were used as dual-functional ions (denoted as COF-S-S-COOH-Ti4+/Nb5+). With the advantage of unbiased enrichment towards phosphopeptides, COF-S-S-COOH-Ti4+/Nb5+ shows ultra-high selectivity (maximum molar ratio of ß-casein: BSA is 1:20,000) and low limit of detection (0.2 fmol). In addition, the material has an excellent phosphopeptide loading capacity (100 µg/mg) and reusability (at least seven times). Furthermore, applying the material to the actual sample, 4 phosphopeptides were selectively extracted from the serum of renal carcinoma patients. At the same time, exosomes with an intact structure in the serum of renal carcinoma patients were successfully isolated rapidly using this strategy. All experiments have shown that COF-S-S-COOH-Ti4+/Nb5+ exhibits exciting potential in practical applications.

Three phases of Gene × Environment interaction research: Theoretical assumptions underlying gene selection.

Some Gene × Environment interaction (G×E) research has focused upon single candidate genes, whereas other related work has targeted multiple genes (e.g., polygenic scores). Each approach has informed efforts to identify individuals who are either especially vulnerable to the negative effects of contextual adversity (diathesis stress) or especially susceptible to both positive and negative contextual conditions (differential susceptibility). A critical step in all such molecular G×E research is the selection of genetic variants thought to moderate environmental influences, a subject that has not received a great deal of attention in critiques of G×E research (beyond the observation of small effects of individual genes). Here we conceptually distinguish three phases of G×E work based on the selection of genes presumed to moderate environmental effects and the theoretical basis of such decisions: (a) single candidate genes, (b) composited (multiple) candidate genes, and (c) GWAS-derived polygenic scores. This illustrative, not exhaustive, review makes it clear that implicit or explicit theoretical assumptions inform gene selection in ways that have not been clearly articulated or fully appreciated.

Differential susceptibility 2.0: Are the same children affected by different experiences and exposures?

Differential susceptibility theory stipulates that some children are more susceptible than others to both supportive and adverse developmental experiences/exposures. What remains unclear is whether the same individuals are most affected by different exposures (i.e., domain general vs. specific). We address this issue empirically for the first time using, for illustrative and proof-of-principle purposes, a novel influence-statistics' method with data from the National Institute of Child Health and Human Development (NICHD) Study of Early Child Care. Results indicated that previously documented effects of greater quality of care on enhanced pre-academic skills and greater quantity of care on more behavior problems apply mostly to different children. Analyses validating the new method indicated, as predicted, that (a) the quantity-of-care effect applied principally to children from more socioeconomically advantaged families and that (b) being highly susceptible to both, one or neither childcare effect varied as a function of a three-gene, polygenic-plasticity score (serotonin transporter linked polymorphic region [5-HTTLPR], dopamine receptor D4 [DRD4], brain-derived neurotrophic factor [BDNF]) in a dose-response manner (i.e., 2>1>0). While domain-specific findings involving child-care effects cannot be generalized to other environmental influences, the influence-statistics' approach appears well suited for investigating the generality-specificity of environment effects, that is, of "differential, differential susceptibility."

Infant temperament, early-childhood parenting, and early-adolescent development: Testing alternative models of Parenting × Temperament interaction.

Here we evaluate whether infant difficult temperament (6 months) functions as a vulnerability or more general plasticity factor when investigating effects of early-childhood parenting (8-42 months) on both positive and negative early-adolescent socioemotional development (age 8-11 years). Using data from the Avon Longitudinal Study of Parents and Children (ALSPAC, N = 14,541) and a re-parameterized model-testing approach to distinguish alternative person × environment conceptual models, results indicated that temperament × parenting interacted in predicting externalizing (i.e., hyperactivity, conduct problems), but not other behavior (i.e., emotional symptoms, peer problems), in a (weak) differential susceptibility manner. While more and less supportive parenting predicted, respectively, fewer and more behavior problems, it did so more strongly for children who were more difficult as infants.

Environmental harshness and unpredictability: Do they affect the same parents and children?

Differential susceptibility theory stipulates that individuals vary in their susceptibility to environmental effects, often implying that the same individuals differ in the same way in their susceptibility to different environmental exposures. The latter point is addressed herein by evaluating the extent to which early-life harshness and unpredictability affect mother's psychological well-being and parenting, as well as their adolescent's life-history strategy, as reflected in number of sexual partners by age 15 years, drawing on data from the National Institute of Child Health and Human Development (NICHD) Study of Early Child Care and Youth Development. Results indicated that mothers whose well-being and parenting proved more susceptible to harshness also proved somewhat more susceptible to environmental unpredictability, with the same being true of adolescent sexual behavior. Nevertheless, findings caution against overgeneralizing sample-level findings to all individuals.

Separation and Purification of Two Saponins from Paris polyphylla var. yunnanensis by a Macroporous Resin.

An effective method for separating and purifying critical saponins (polyphyllin II and polyphyllin VII) from a Paris polyphylla var. yunnanensis extract was developed in this study which was environmentally friendly and economical. Static adsorption kinetics, thermodynamics, and the dynamic adsorption-desorption of macroporous resins were investigated, and then the conditions of purification and separation were optimized by fitting with an adsorption thermodynamics equation and a kinetic equation. Effective NKA-9 resin from seven macroporous resins was screened out to separate and purify the two saponins. The static adsorption and dynamic adsorption were chemical and physical adsorption dual-processes on the NKA-9 resin. Under the optimum parameters, the contents of polyphyllin II and polyphyllin VII in the product were 17.3-fold and 28.6-fold those in plant extracts, respectively. The total yields of the two saponins were 93.16%. This research thus provides a theoretical foundation for the large-scale industrial production of the natural drugs polyphyllin II and polyphyllin VII.