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Autism spectrum disorder (ASD) is characterized by atypical communication, social interactions, and restricted interests. In ASD, there are dysfunctional immune regulatory control mechanisms that can lead to immune activation. Notably lower frequencies of regulatory T cells (Tregs) and reduced immunosuppressive cytokines are reported and associated with more impaired behaviors impacting both individuals with autism and their families. Therefore, therapeutic approaches that enhance immune regulation may offer substantial benefits. Using the maternal immune activation (MIA) model, we investigated whether adoptive transfer of wildtype Tregs into MIA offspring recipients could rescue immune activation, brain transcriptome changes and behaviors exhibited in adult MIA offspring. We also aimed to explore potential sex-differences in responses. In male but not female MIA offspring, Tregs transfer reduced the frequency of T helper (T _H )-17 (RORγT⁺ CD4⁺) T cells in both the mesenteric lymph node (MLN) and spleen. Moreover, the frequency of CD25⁺Foxp3⁺ T cells was increased in the MLN and spleen of male but not female MIA offspring following Treg transfer. Splenocytes from male MIA offspring receiving Tregs showed reduced production of inflammatory cytokines (e.g., IL-6 and TNFα) following PMA/Ionomycin stimulations. In contrast, female MIA offspring that received Tregs exhibited different cytokine profiles characterized by increased production of cytokines, including GM-CSF, IFNγ, and IL-10. In the brain, bulk mRNA sequencing in the cerebellum, frontal cortex, and hippocampus revealed that Tregs-treated male MIA offspring had differentially expressed genes involved in neurodevelopmental disorders, synaptic function, and epigenetic regulation. Minimal gene expression differences were observed in female counterparts. There was significant improvement in self-grooming behaviors in males MIA offspring that received Tregs. In females, social novelty improved after Tregs treatment. In summary, adoptive Tregs transfer reduced systemic inflammation, brain transcription and behavior alterations in a sex dependent manner in the context of MIA. These findings suggest that adoptive Treg transfer may represent a viable therapeutic avenue for mitigating systemic inflammation and comorbidities associated with MIA and neurodevelopmental disorders such as ASD.

Background: Although hypermobile Ehlers-Danlos syndrome (hEDS) and hypermobility spectrum disorders (HSD) are inherited disorders expected to display a 1:1 sex ratio, most studies report a strong female dominance. This study analyzed sex differences in 122 self-reported symptoms and comorbidities in patients diagnosed with hEDS or HSD using the 2017 criteria. Methods: Self-reported data was obtained from November 1, 2019, to April 25, 2025, ( n =2,451) from an Intake Questionnaire of adult (≤18 years old) patients diagnosed with hEDS or HSD at the EDS Clinic at Mayo Clinic Florida and analyzed by sex. Results: We found that 90.6% ( n =575) of hEDS patients were female and 9.4% ( n =60) were male, with a sex ratio of 9.6:1 female-to-male, while 95.2% ( n =1,728) of HSD patients were female and 4.8% ( n =88) were male, with a sex ratio of 19.6:1 female-to-male. Females were older than males: hEDS 35 vs. 28 years of age ( p <0.001), HSD 34 vs. 31 years of age ( p =0.004). Overall, females self-reported more symptoms/comorbidities than males: hEDS 31/122 (25.4%), HSD 59/122 (48.4%). In contrast, only 5/122 (4.1%) symptoms/comorbidities were self-reported more often in males including attention deficit disorder/attention deficit hyperactivity disorder, delay in developmental milestones, snoring, autism spectrum disorder, and obstructive sleep apnea. Females with HSD had more mast cell-related symptoms (i.e., hives p <0.001) which were reflected in higher mast cell scores than males ( p <0.001). Patients with higher mast cell scores also had higher serum levels of total IgE ( p =0.029). More females with HSD were diagnosed with fibromyalgia ( p <0.001) and reported symptoms associated with autonomic dysfunction than males. Aside from abuse, which was higher in females (hEDS p =0.039, HSD p <0.001), few sex differences were reported for psychological symptoms/comorbidities. In support for the idea that elevated mast cell activity in females may lead to extracellular matrix remodeling that promotes hypermobility, females with hEDS/HSD had greater serum collagen-4α1 ( p <0.0001) and fibronectin ( p =0.015) than males by ELISA. Conclusions: Females with hEDS and HSD report a higher burden of symptoms/comorbidities than males, providing a possible explanation for fewer males being reported in demographic data. Sex differences in symptoms/comorbidities may reflect sex differences in pathogenic drivers of disease.

Current cell transfection systems face efficiency limitations, particularly for challenging cell types. We present a sponge-assisted transfection method enabling efficient mRNA transfection at nanogram-scale doses-a several orders of magnitude-reduction compared to the microgram quantities typically required by lipid-based and membrane-disruption methods. This approach achieves robust, dose-dependent transfection across difficult-to-transfect cell types, establishing broad applicability for both basic research and therapeutic use.

The histone variant H2A.Z resides within active gene promoters, but how it influences chromatin state and nucleosome stability remains poorly understood. Here, we probe two H2A.Z binding proteins with seemingly opposing function: VPS72, an SRCAP subunit that aids in H2A.Z deposition, and ANP32E, a histone chaperone which is thought to remove H2A.Z. Using functional genomics and biochemical assays, we show that VPS72 and ANP32E co-occupy active promoters yet function antagonistically. VPS72 promotes H2A.Z incorporation, acetylation, BRG1 recruitment, and transcription, whereas ANP32E constrains these features by stabilizing nucleosomes. Loss of ANP32E increases VPS72 binding, chromatin accessibility, and transcription, while co-depletion of VPS72 reverses these effects. Reconstitution assays show that ANP32E promotes nucleosome assembly and prevents DNA unwrapping, providing a mechanistic basis for in vivo function. Our results support a model where promoter accessibility arises from antagonistic and cooperative actions, revealing a general paradigm in which counterbalancing factors govern gene regulatory potential.

TFIID is instrumental in recognizing promoter sequences and initiating transcription, yet a cohesive understanding of how this complex interacts with and functions at different promoter types in vivo is still lacking. Here, we employed ChIP-nexus to capture high resolution binding footprints of all Drosophila TFIID subunits across the genome. These footprints reveal TFIID sub-modules whose DNA contacts suggest new structural details. At different promoter types, the footprints of the TAFs are very similar, suggesting the presence of engaged TFIID across all promoters. In contrast, the binding profile of TBP is promoter-specific, enabling us to identify TATA, DPR, and TCT/housekeeping promoters de novo, along with their underlying core promoter elements. Notably, our data point to NC2 being specific for TBP binding at the TATA box and suggest that TATA promoters show both TAF-dependent and TAF-independent initiation in vivo. These data suggest a model for the increased burst size observed at TATA promoters and provide a comprehensive resource for linking structural and biochemical results to in vivo data.

We tend to think of neurons as either excitatory or inhibitory, but certain neurons chemically inhibit their downstream targets while electrically exciting their neighbors. For example, in the cerebellum, molecular layer interneurons (MLIs) inhibit Purkinje cells (P-cells) via release of GABA but promote spiking in each other via gap junctions. What is gained by having an inhibitory neuron excite its neighbor? Here, we recorded activities of P-cells and MLIs as marmosets performed saccadic eye movements and found that spike timing in pairs of neighboring neurons of the same type exhibited a mathematical regularity: as firing rates increased, rate of spikes that were within 1ms of each other grew disproportionately while 2-4ms intervals were suppressed. To uncover the purpose of this coordination, during saccades we recorded thousands of neuron triplets in which two MLIs converged onto a single target P-cell. When the MLIs spiked within 1ms of each other, they produced superposition of their individual effects on their target; a deep inhibition followed by a post-inhibitory rebound. However, when the MLIs spiked 2-4ms apart, the two spikes interfered with each other, producing partial cancellation. Thus, electrical coupling between inhibitory neurons orchestrated their spike timing so that as firing rates increased, the temporal intervals that induced constructive superposition were promoted while the intervals that caused destructive competition were suppressed.

Background In low malaria transmission settings targeting elimination, the World Health Organization recommends a combination of mass (e.g., mass test-and-treat), targeted (e.g., chemoprophylaxis or treatment for travelers), and reactive (e.g., reactive drug administration) strategies. Most of these strategies would not identify and treat individuals with asymptomatic parasitemia. This study was conducted in a pre-elimination setting in Southern Province, Zambia to examine risk factors for asymptomatic parasitemia, its epidemiologic relationship to incident clinical malaria, and evidence of its contribution to ongoing transmission to inform policy on whether these parasitemic individuals need to be identified and treated to achieve malaria elimination. Methods An intensive longitudinal cohort study of 197 households within the catchment area of a single health center was designed to capture all clinical malaria cases and episodes of asymptomatic parasitemia between 2018 and 2020. During monthly collections, all household members and overnight visitors were administered a questionnaire and a blood sample was collected to identify Plasmodium falciparum parasitemia by qPCR. Passive surveillance was also established at the local health center to identify cases of clinical malaria. The statistical analysis examined risk factors for parasitemia and associations between asymptomatic parasitemia and subsequent episodes of clinical malaria within individuals and parasitemia in household members. Results Of the 1071 individuals enrolled in the cohort, 144 (13%) were positive by qPCR for P. falciparum at least once during the two-year study period. Monthly parasite prevalence by qPCR never exceeded 4% and parasite density was very low with a median of four parasites/µL. Incidence of self-reported clinical malaria was 46.7 cases per 1000 person-years. Low-density asymptomatic parasitemia was identified in all age groups, including young children. There was no association between asymptomatic parasitemia and clinical malaria within individuals, nor was there an association between asymptomatic parasitemia and subsequent parasitemia in household members beyond one month of the index case. Conclusion Low-level parasitemia was prevalent despite few cases of clinical malaria in this low transmission setting. There was no evidence that low-level asymptomatic parasitemia led to clinical cases of malaria or transmission to other household members.

Objective To estimate cervical cancer screening rates, prevalence of risk factors, and factors associated with being overdue for screening among people experiencing homelessness in two Indiana cities. Methods Rapid assessment surveys were conducted at two large homelessness service agencies in Indianapolis and Lafayette, Indiana (November 2023 to November 2024). Participants were aged 21-69 years, assigned female at birth, and currently experiencing homelessness. Screening status was categorized as up-to-date (screened within 5 years) or overdue (more than 5 years or never screened). Descriptive statistics and multivariable logistic regression examined predictors of being overdue. Results Among n = 212 participants, 35% were overdue and 49% had not been screened within 3 years. Prevalence of risk factors was high, including smoking (74%), sexual debut before age 18 (73%), and no HPV vaccination (75%). Older age and having experienced homelessness for 5 years or more were associated with higher odds of being overdue. Conclusions Longer duration of homelessness significantly increased the likelihood of being overdue for cervical cancer screening, underscoring the cumulative disadvantage of chronic housing instability. Cervical cancer prevention is a critical unmet need among women experiencing homelessness, and findings highlight the importance of developing targeted interventions to improve screening access in this population.

Primary liver cancers, including hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), arise from the neoplastic transformation of hepatocytes and cholangiocytes, respectively. Loss or downregulation of PTEN, a tumor suppressor negatively regulating the PI3K/AKT pathway, is frequently observed in CCA and HCC. Notably, PTEN mutations are observed at nearly twice the frequency in combined CCA-HCC tumors than either HCC or CCA alone. Using lineage-specific liver-targeted PTEN-deficient mouse models, we demonstrate that PTEN loss drives cellular dedifferentiation and tumorigenesis, a process that is critically dependent on AKT2. Mechanistically, PTEN deficiency induces activation of NOTCH and upregulation of transcriptional factor SOX9, which plays a central role in tumor cell transformation. In parallel, PTEN loss increases SMAD4 expression and sensitizes the tumor cells to TGFβ signaling, with TGFβ treatment repressing SOX9 expression in tumor cells lacking PTEN. Together, our study defined a critical role for PTEN-AKT2 signaling in maintaining liver epithelial lineage fidelity and revealed how its disruption promotes the conversion of mature hepatocytes or cholangiocytes into liver cancer stem-like cells (LCSCs). Furthermore, we identify a PTEN-dependent crosstalk between NOTCH and TGFβ pathways that governs liver tumor development. Together, this work provides mechanistic insight into lineage plasticity in liver cancer with implications for pathway-directed therapy.