Applying the Discrepancy Consistency Method on CAS-2: Brief Data in a Sample of Greek-Speaking Children.

This study aimed to examine whether we could use the discrepancy consistency method on CAS-2: Brief data collected in Cyprus. A total of 438 Grade 6 children (201 boys, 237 girls, Mage = 135.75 months, SD = 4.05 months) from Cyprus were assessed on the Cognitive Assessment System-2: Brief that is used to operationalize four neurocognitive processes, namely Planning, Attention, Simultaneous, and Successive (PASS) processing. They were also assessed on two measures of reading (Wordchains and CBM-Maze) and mathematics (Mathematics Achievement Test and Mathematics Reasoning Test). The results showed that 31.5% of our sample had a PASS disorder, and 8% to 10% of our sample had both a PASS disorder and an academic disorder. These numbers are similar to those reported in previous studies that used DCM in North America and suggest that the method can be used to inform instruction, particularly in places where no screening for learning disabilities is available.

Hybrid immunity to SARS-CoV-2 arises from serological recall of IgG antibodies distinctly imprinted by infection or vaccination.

We used plasma IgG proteomics to study the molecular composition and temporal durability of polyclonal IgG antibodies triggered by ancestral SARS-CoV-2 infection, vaccination, or their combination ("hybrid immunity"). Infection, whether primary or post-vaccination, mainly triggered an anti-spike antibody response to the S2 domain, while vaccination predominantly induced anti-RBD antibodies. Immunological imprinting persisted after a secondary (hybrid) exposure, with >60% of the ensuing serological response originating from the initial antibodies generated during the first exposure. We highlight one instance where hybrid immunity arising from breakthrough infection resulted in a marked increase in the breadth and affinity of a highly abundant vaccination-elicited plasma IgG antibody, SC27. With an intrinsic binding affinity surpassing a theoretical maximum (K D < 5 pM), SC27 demonstrated potent neutralization of various SARS-CoV-2 variants and SARS-like zoonotic viruses (IC 50 ∼0.1-1.75 nM) and provided robust protection in vivo . Cryo-EM structural analysis unveiled that SC27 binds to the RBD class 1/4 epitope, with both VH and VL significantly contributing to the binding interface. These findings suggest that exceptionally broad and potent antibodies can be prevalent in plasma and can largely dictate the nature of serological neutralization. HIGHLIGHTS: ▪ Infection and vaccination elicit unique IgG antibody profiles at the molecular level▪ Immunological imprinting varies between infection (S2/NTD) and vaccination (RBD)▪ Hybrid immunity maintains the imprint of first infection or first vaccination▪ Hybrid immune IgG plasma mAbs have superior neutralization potency and breadth.

Human Tumor-Associated Macrophages and Neutrophils Regulate Antitumor Antibody Efficacy through Lethal and Sublethal Trogocytosis.

The clinical benefits of tumor-targeting antibodies (tAb) are modest in solid human tumors. The efficacy of many tAbs is dependent on Fc receptor (FcR)-expressing leukocytes that bind Fc fragments of tAb. Tumor-associated macrophages (TAM) and neutrophils (TAN) represent the majority of FcR+ effectors in solid tumors. A better understanding of the mechanisms by which TAMs and TANs regulate tAb response could help improve the efficacy of cancer treatments. Here, we found that myeloid effectors interacting with tAb-opsonized lung cancer cells used antibody-dependent trogocytosis (ADT) but not antibody-dependent phagocytosis. During this process, myeloid cells "nibbled off" tumor cell fragments containing tAb/targeted antigen (tAg) complexes. ADT was only tumoricidal when the tumor cells expressed high levels of tAg and the effectors were present at high effector-to-tumor ratios. If either of these conditions were not met, which is typical for solid tumors, ADT was sublethal. Sublethal ADT, mainly mediated by CD32hiCD64hi TAM, led to two outcomes: (i) removal of surface tAg/tAb complexes from the tumor that facilitated tumor cell escape from the tumoricidal effects of tAb; and (ii) acquisition of bystander tAgs by TAM with subsequent cross-presentation and stimulation of tumor-specific T-cell responses. CD89hiCD32loCD64lo peripheral blood neutrophils (PBN) and TAN stimulated tumor cell growth in the presence of the IgG1 anti-EGFR Ab cetuximab; however, IgA anti-EGFR Abs triggered the tumoricidal activity of PBN and negated the stimulatory effect of TAN. Overall, this study provides insights into the mechanisms by which myeloid effectors mediate tumor cell killing or resistance during tAb therapy. SIGNIFICANCE: The elucidation of the conditions and mechanisms by which human FcR+ myeloid effectors mediate cancer cell resistance and killing during antibody treatment could help develop improved strategies for treating solid tumors.

Contrasting Direct Instruction in Morphological Decoding and Morphological Inquiry-Analysis Interventions in Grade 3 Children With Poor Morphological Awareness.

We report on a school-based randomized control trial study comparing two morphological interventions with untaught controls: one focusing on direct instruction targeting print morphological decoding (direct decoding condition) and the other on inquiry-focused pedagogy using oral morphological analysis (inquiry-analysis condition). We identified 63 Grade 3 children with below-average morphological awareness following screening (from N = 163). This sub-sample showed average pseudoword decoding but poor language and word reading abilities. Following a 13-week supplemental intervention randomized within the 63 children, results showed a statistically significant main effect of intervention on standardized reading vocabulary measures at immediate post-test in the direct decoding condition. Pre-test morphological awareness moderated reading vocabulary effects for the untaught control group. Statistically significant moderation of growth in sentence comprehension at post- by pre-test morphological awareness was also evident in the inquiry-analysis condition. Universal screening for below-average morphological awareness followed by inquiry-based or direct instruction interventions focusing on the meaning dimensions of morphemes may be modestly efficacious for supporting reading vocabulary and sentence comprehension in such at risk learners, potentially aiding school-wide literacy improvement.

Internalizing problems in individuals with reading, mathematics and unspecified learning difficulties: a systematic review and meta-analysis.

We conducted a systematic review and meta-analysis to examine if individuals with reading difficulties (RD), mathematics difficulties (MD), or unspecified learning difficulties (ULD) experience internalizing problems (i.e., anxiety, depression, somatic complaints, and social withdrawal) to the same extent, and if the effect sizes are influenced by moderators (age, internalizing problems type, anxiety type, rater type, selection criteria, and attention control). We reviewed 2,806 studies published in English between January 2000 and April 2023. Our final sample consisted of 96 studies that reported effect sizes or data to calculate them (a total of 120 unique samples, 83,260 participants, age range = 7.3 - 34.8 years). Risk of bias and sensitivity were assessed. A random-effects model analysis revealed a significant and moderate overall effect size (Hedge's g = -.54), indicating that individuals with RD, MD or ULD experience more internalizing problems than their chronological-age (CA) controls. Follow-up analyses showed that neither learning difficulties type nor age of participants were significant moderators, but selection criteria (diagnosis versus screening) and internalizing problems type were. These findings suggest that individuals with RD do not differ from those with MD or ULD in internalizing problems, and all fare poorly compared to CA controls.

SARS-COV-2 Omicron variants conformationally escape a rare quaternary antibody binding mode.

The ongoing evolution of SARS-CoV-2 into more easily transmissible and infectious variants has provided unprecedented insight into mutations enabling immune escape. Understanding how these mutations affect the dynamics of antibody-antigen interactions is crucial to the development of broadly protective antibodies and vaccines. Here we report the characterization of a potent neutralizing antibody (N3-1) identified from a COVID-19 patient during the first disease wave. Cryogenic electron microscopy revealed a quaternary binding mode that enables direct interactions with all three receptor-binding domains of the spike protein trimer, resulting in extraordinary avidity and potent neutralization of all major variants of concern until the emergence of Omicron. Structure-based rational design of N3-1 mutants improved binding to all Omicron variants but only partially restored neutralization of the conformationally distinct Omicron BA.1. This study provides new insights into immune evasion through changes in spike protein dynamics and highlights considerations for future conformationally biased multivalent vaccine designs.

Dynamics and durability of HIV-1 neutralization are determined by viral replication.

Human immunodeficiency virus type 1 (HIV-1)-neutralizing antibodies (nAbs) that prevent infection are the main goal of HIV vaccine discovery. But as no nAb-eliciting vaccines are yet available, only data from HIV-1 neutralizers-persons with HIV-1 who naturally develop broad and potent nAbs-can inform about the dynamics and durability of nAb responses in humans, knowledge which is crucial for the design of future HIV-1 vaccine regimens. To address this, we assessed HIV-1-neutralizing immunoglobulin G (IgG) from 2,354 persons with HIV-1 on or off antiretroviral therapy (ART). Infection with non-clade B viruses, CD4+ T cell counts <200 µl-1, being off ART and a longer time off ART were independent predictors of a more potent and broad neutralization. In longitudinal analyses, we found nAb half-lives of 9.3 and 16.9 years in individuals with no- or low-level viremia, respectively, and 4.0 years in persons who newly initiated ART. Finally, in a potent HIV-1 neutralizer, we identified lower fractions of serum nAbs and of nAb-encoding memory B cells after ART initiation, suggesting that a decreasing neutralizing serum activity after antigen withdrawal is due to lower levels of nAbs. These results collectively show that HIV-1-neutralizing responses can persist for several years, even at low antigen levels, suggesting that an HIV-1 vaccine may elicit a durable nAb response.

Enhanced SARS-CoV-2 humoral immunity following breakthrough infection builds upon the preexisting memory B cell pool.

The human immune response must continuously adapt to newly emerging SARS-CoV-2 variants. To investigate how B cells respond to repeated SARS-CoV-2 antigen exposure by Wu01 booster vaccination and Omicron breakthrough infection, we performed a molecular longitudinal analysis of the memory B cell pool. We demonstrate that a subsequent breakthrough infection substantially increases the frequency of B cells encoding SARS-CoV-2-neutralizing antibodies. However, this is not primarily attributable to maturation, but to selection of preexisting B cell clones. Moreover, broadly reactive memory B cells arose early and even neutralized highly mutated variants like XBB.1.5 that the individuals had not encountered. Together, our data show that SARS-CoV-2 immunity is largely imprinted on Wu01 over the course of multiple antigen contacts but can respond to new variants through preexisting diversity.

Peripheral T cell activation, not thymic selection, expands the T follicular helper repertoire in a lupus-prone murine model.

Many autoimmune diseases are characterized by the activation of autoreactive T cells. The T cell repertoire is established in the thymus; it remains uncertain whether the presence of disease-associated autoreactive T cells reflects abnormal T cell selection in the thymus or aberrant T cell activation in the periphery. Here, we describe T cell selection, activation, and T cell repertoire diversity in female mice deficient for B lymphocyte-induced maturation protein (BLIMP)-1 in dendritic cells (DCs) (Prdm1 CKO). These mice exhibit a lupus-like phenotype with an expanded population of T follicular helper (Tfh) cells having a more diverse T cell receptor (TCR) repertoire than wild-type mice and, in turn, develop a lupus-like pathology. To understand the origin of the aberrant Tfh population, we analyzed the TCR repertoire of thymocytes and naive CD4 T cells from Prdm1 CKO mice. We show that early development and selection of T cells in the thymus are not affected. Importantly, however, we observed increased TCR signal strength and increased proliferation of naive T cells cultured in vitro with antigen and BLIMP1-deficient DCs compared to control DCs. Moreover, there was increased diversity in the TCR repertoire in naive CD4+ T cells stimulated in vitro with BLIMP1-deficient DCs. Collectively, our data indicate that lowering the threshold for peripheral T cell activation without altering thymic selection and naive T cell TCR repertoire leads to an expanded repertoire of antigen-activated T cells and impairs peripheral T cell tolerance.

Do Performance-Based Measures and Behavioral Ratings of Executive Functioning Complement Each Other in Predicting Reading and Mathematics in Chinese?

We examined what executive functioning (EF) components predict reading and mathematics within the same study and whether the effects of behavioral ratings of EF overlap or complement those of performance-based measures. One hundred and nine Grade 2 Mandarin-speaking Chinese students from Chengdu, China (55 girls, 54 boys, Mage = 8.15 years), were assessed on measures of EF (planning, inhibition, shifting, and working memory), speed of processing, reading and mathematics. Parents also rated their children's EF skills using the Childhood Executive Functioning Inventory. Results of hierarchical regression analyses showed that only working memory among the performance-based EF measures predicted reading and mathematics. In addition, none of the behavioral ratings of EF made a significant contribution to reading and mathematics after controlling for mother's education and speed of processing. Taken together, these findings suggest that working memory is a domain general predictor of academic achievement, but only when measured with cognitive tasks.

Domain Specific and Cross Domain Associations between PASS Cognitive Processes and Academic Achievement.

The purpose of this study was to examine the role of intelligence-operationalized in terms of Planning, Attention, Simultaneous, and Successive (PASS) processing skills-in reading and mathematics. Two hundred and forty-two Grade 6 Greek-speaking students (114 boys and 128 girls, Mage = 135.65 months, SD = 4.12 months) were assessed on PASS processes, speed of processing (Visual Matching), reading (Wordchains and CBM-Maze), and mathematics (Mathematics Achievement Test and Mathematics Reasoning Test). The results of the hierarchical regression analyses showed that, after controlling for family's socioeconomic status and speed of processing, Attention and Successive processing predicted reading and Planning and Simultaneous processing predicted mathematics. Taken together, these findings suggest that different PASS processes may account for individual differences in reading and mathematics.

What Components of Working Memory Are Impaired in Children with Reading and/or Mathematics Difficulties?

Both reading difficulties (RD) and mathematics difficulties (MD) are common neurodevelopmental disorders. The co-occurrence of RD and MD, known as comorbid RDMD, is estimated to range between 21% and 45% of children with learning disabilities. Deficits in working memory have been reported in both RD and MD groups, as well as among comorbid RDMD. However, previous comorbidity studies have only examined the role of some components of working memory, and they do not strictly match their groups on relevant reading and mathematics tasks. Thus, the purpose of this study is to examine the nature of working memory deficits in comorbid RDMD after matching groups based on reading and mathematics tasks. We assessed four groups of children (RD [n = 21, Mage = 10.96 years], MD [n = 24, Mage = 11.04 years], comorbid RDMD [n = 26, Mage = 10.90 years], and chronological-age controls [n = 27, Mage = 10.96 years]) on measures of the phonological loop (word span and digit span forward tasks), central executive (complex word and digit span), and updating tasks (word and digit 2-back). The results of ANCOVA (covarying for gender and non-verbal IQ) showed first that the RD and RDMD groups performed significantly worse than the MD and control groups in both measures of the phonological loop. For the central executive and updating tasks, we found an effect based on stimulus type. For word-related tasks, the RD and comorbid RDMD groups performed worse than the MD and control groups, and for number-related tasks, the MD and comorbid RDMD groups performed worse than the RD and control groups. Taken together, our findings provide support for the correlated liability model of comorbidity, which indicates that working memory deficits experienced by the RDMD group are an additive combination of deficits observed in the RD and MD groups, suggesting that working memory tasks used to examine underlying deficits in reading and/or mathematics difficulties may dictate whether or not significant group differences are found.

Enzyme-mediated depletion of methylthioadenosine restores T cell function in MTAP-deficient tumors and reverses immunotherapy resistance.

Chromosomal region 9p21 containing tumor suppressors CDKN2A/B and methylthioadenosine phosphorylase (MTAP) is one of the most frequent genetic deletions in cancer. 9p21 loss is correlated with reduced tumor-infiltrating lymphocytes (TILs) and resistance to immune checkpoint inhibitor (ICI) therapy. Previously thought to be caused by CDKN2A/B loss, we now show that it is loss of MTAP that leads to poor outcomes on ICI therapy and reduced TIL density. MTAP loss causes accumulation of methylthioadenosine (MTA) both intracellularly and extracellularly and profoundly impairs T cell function via the inhibition of protein arginine methyltransferase 5 (PRMT5) and by adenosine receptor agonism. Administration of MTA-depleting enzymes reverses this immunosuppressive effect, increasing TILs and drastically impairing tumor growth and importantly, synergizes well with ICI therapy. As several studies have shown ICI resistance in 9p21/MTAP null/low patients, we propose that MTA degrading therapeutics may have substantial therapeutic benefit in these patients by enhancing ICI effectiveness.

Association of the Bacteria of the Vermiform Appendix and the Peritoneal Cavity with Complicated Acute Appendicitis in Children.

BACKGROUND: Primary infection has been questioned as the pathogenetic cause of acute appendicitis. We attempted to identify the bacteria involved and to investigate if their species, types, or combinations affected the severity of acute appendicitis in children. METHODS: Samples from both the appendiceal lumen and the peritoneal cavity of 72 children who underwent appendectomy were collected to perform bacterial culture analysis. The outcomes were studied to identify if and how they were associated with the severity of the disease. Regression analysis was performed to identify any risk factors associated with complicated appendicitis. RESULTS: Escherichia coli, Pseudomonas aeruginosa, and Streptococcus species were the most common pathogens found in the study population. The same microorganisms, either combined or separate, were the most common in the appendiceal lumen and the peritoneal cavity of patients with complicated appendicitis. Gram-negative bacteria and polymicrobial cultures in the peritoneal fluid and in the appendiceal lumen were associated with complicated appendicitis. Polymicrobial cultures in the peritoneal cavity presented a four times higher risk of complicated appendicitis. CONCLUSIONS: Polymicrobial presentation and Gram-negative bacteria are associated with complicated appendicitis. Antibiotic regimens should target the combinations of the most frequently identified pathogens, speculating the value of early antipseudomonal intervention.

Enzymatic depletion of l-Met using an engineered human enzyme as a novel therapeutic strategy for melanoma.

Many cancers, including melanoma, have a higher requirement for l-methionine in comparison with noncancerous cells. In this study, we show that administration of an engineered human methionine-γ-lyase (hMGL) significantly reduced the survival of both human and mouse melanoma cells in vitro. A multiomics approach was utilized to identify global changes in gene expression and in metabolite levels with hMGL treatment in melanoma cells. There was considerable overlap in the perturbed pathways identified in the two data sets. Common pathways were flagged for further investigation to understand their mechanistic importance. In this regard, hMGL treatment induced S and G2 phase cell cycle arrest, decreased nucleotide levels, and increased DNA double-strand breaks suggesting an important role for replication stress in the mechanism of hMGL effects on melanoma cells. Further, hMGL treatment resulted in increased cellular reactive oxygen species levels and increased apoptosis as well as uncharged transfer RNA pathway upregulation. Finally, treatment with hMGL significantly inhibited the growth of both mouse and human melanoma cells in orthotopic tumor models in vivo. Overall, the results of this study provide a strong rationale for further mechanistic evaluation and clinical development of hMGL for the treatment of melanoma skin cancer and other cancers.

Cysteine depletion sensitizes prostate cancer cells to agents that enhance DNA damage and to immune checkpoint inhibition.

BACKGROUND: Prostate Cancer (PCa) represents one of the most commonly diagnosed neoplasms in men and is associated with significant morbidity and mortality. Therapy resistance and significant side effects of current treatment strategies indicate the need for more effective agents to treat both androgen-dependent and androgen-independent PCa. In earlier studies, we demonstrated that depletion of L-cysteine/cystine with an engineered human enzyme, Cyst(e)inase, increased intracellular ROS levels and inhibited PCa growth in vitro and in vivo. The current study was conducted to further explore the mechanisms and potential combinatorial approaches with Cyst(e)inase for treatment of PCa. METHODS: DNA single strand breaks and clustered oxidative DNA damage were evaluated by alkaline comet assay and pulsed field gel electrophoresis, respectively. Neutral comet assay and immunofluorescence staining was used to measure DNA double strand breaks. Cell survival and reactive oxygen species level were measured by crystal violet assay and DCFDA staining, respectively. Western blot was used to determine protein expression. FACS analyses were preformed for immune cell phenotyping. Allograft and xenograft tumor models were used for assessing effects on tumor growth. RESULTS: PCa cells treated with Cyst(e)inase lead to DNA single and double strand breaks resulted from clustered oxidative DNA damage (SSBs and DSBs). Cyst(e)inase in combination with Auranofin, a thioredoxin reductase inhibitor, further increased intracellular ROS and DNA DSBs and synergistically inhibited PCa cell growth in vitro and in vivo. A combination of Cyst(e)inase with a PARP inhibitor (Olaparib) also increased DNA DSBs and synergistically inhibited PCa cell growth in vitro and in vivo without additional ROS induction. Knockdown of BRCA2 in PCa cells increased DSBs and enhanced sensitivity to Cyst(e)inase. Finally, Cyst(e)inase treatment altered tumor immune infiltrates and PD-L1 expression and sensitized PCa cells to anti-PD-L1 treatment. CONCLUSIONS: The current results demonstrate the importance of oxidative DNA damage either alone or in combination for Cyst(e)inase-induced anticancer activity. Furthermore, cysteine/cystine depletion alters the tumor immune landscape favoring enhanced immune checkpoint inhibition targeting PD-L1. Thus, combinatorial approaches with Cyst(e)inase could lead to novel therapeutic strategies for PCa.

Serial and discrete naming and reading in Chinese first graders: Testing predictions from the cascaded processing hypothesis.

Recent studies have suggested that-beyond automaticity and prosody-reading fluency involves parallel processing of adjacent items presented in a sequence, termed "cascaded processing." To date, most studies examining cascaded processing have been conducted in alphabetic orthographies. Thus, the purpose of this study was to examine the cascaded processing hypothesis in Chinese. A total of 119 Grade 1 Chinese children (61 boys and 58 girls; Mage = 7.30 years, SD = 0.31) were assessed on serial and discrete naming of digits as well as on serial and discrete naming of high-frequency one- and two-character words and low-frequency one-character words presented with pinyin. Results of hierarchical regression analyses showed, first, that serial digit naming was a unique predictor of discrete naming of low-frequency one-character words and two-character words, but not of high-frequency one-character words. Second, serial digit naming was a unique predictor of reading of high-frequency one- and two-character word reading after controlling for discrete word reading. These findings suggest that Chinese first graders process high-frequency characters holistically (similar to simple digits), which then facilitates parallel processing of multiple stimuli when they are presented in a sequence.

Reading Fluency in Chinese Children With Reading Disabilities and/or ADHD: A Key Role for Morphology.

The Triangle Model of Reading proposes that phonology, orthography, and semantics are crucial to understand word reading and reading disability (RD). Morphology has been added as a binding agent to this model. However, it is unclear how these variables relate to word reading in children with attention deficit/hyperactivity disorder (ADHD) or comorbid ADHD and RD (ADHD+RD). This study examined the performance of Chinese children with RD, ADHD, or ADHD+RD in phonology, orthography, semantics, and morphology, and investigated whether morphology made an additional contribution beyond the other skills in explaining word reading fluency. Participants were 151 Grade 1 to 3 Chinese students: RD (n = 31), ADHD (n = 43), ADHD+RD (n = 27), and typically developing controls (TD, n = 50). Results indicated that children with ADHD+RD (a) showed similar performance to RD and ADHD in tone awareness, orthographic legality, and homophone morpheme awareness; (b) had similar performance to RD but worse than ADHD in phonology, semantics, and morpheme production; and (c) had more severe deficits than RD and ADHD in orthographic reversal, morpheme identification, and homograph awareness. Morphology significantly predicted word reading fluency beyond the other skills, and its predictive effect was more salient for ADHD+RD, ADHD, and TD. The findings provide evidence of both shared and additive effects of RD and ADHD. Morphology may be an important diagnostic factor in identifying Chinese reading and behavioral deficit groups and a worthwhile target for intervention.