Wisconsin National Primate Research Center

Purpose The kappa opioid receptor (KOR) plays a pivotal role in stress- and anxiety-related behaviors, with growing evidence linking it to stress induced by social isolation and separation. Despite this, tools for studying KOR in clinically relevant social contexts remain limited. The socially monogamous coppery titi monkey offers a translational model for investigating pair bonding. This study evaluated the feasibility of [ ¹¹ C]GR103545 PET imaging to characterize KOR activity in vivo , and its pharmacological blockade for the first time in titi monkeys. Methods Adult titi monkeys (N = 6) underwent [ ¹¹ C]GR103545 PET brain scans at baseline and following administration of the KOR antagonist CERC-501. Non-displaceable binding potential (BP ND ) was calculated across 14 brain volumes of interest (VOIs) implicated in social bonding, using Simplified and Logan reference tissue models (SRTM and LRTM), with the cerebellum as the reference region. Results Baseline [ ¹¹ C]GR103545 uptake patterns across VOIs were consistent with reports in humans, other primates and published autoradiography data. CERC-501 pretreatment significantly reduced BP ND (SRTM: 55.99%, LRTM: 59.68%) across several, but not all brain VOIs. Conclusions This study establishes [ ¹¹ C]GR103545 PET as a viable tool for assessing KOR binding dynamics in titi monkeys, providing new opportunities to explore KOR modulation in social bonding and separation.

The human genome contains millions of copies of retrotransposons that are silenced but many of these copies have potential to become active if mutated, having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affect their jumping activity. Here, we develop a massively parallel jumping assay (MPJA) that tests the jumping potential of thousands of transposons en masse. We generate a nucleotide variant library of four Alu retrotransposons containing 165,087 different haplotypes and test them for their jumping ability using MPJA. We found 66,821 unique jumping haplotypes, allowing us to pinpoint domains and variants vital for transposition. Mapping these variants to the Alu-RNA secondary structure revealed stem-loop features that contribute to jumping potential. Combined, our work provides a high-throughput assay that assesses the ability of retrotransposons to jump and identifies nucleotide changes that have the potential to reactivate them in the human genome.

Cervical mucus changes throughout the menstrual cycle in response to hormonal fluctuations, regulating access of sperm and pathogens to the reproductive tract. CFTR is an anion channel that plays a critical role in mediating epithelial mucus secretions. Primary endocervical cells obtained from rhesus macaques Macaca mulatta were cultured using conditional reprogramming and treated with vehicle controls or CFTR inhibitors. In order to measure changes in hydration and viscosity of secreted mucus, we adapted two airway mucus assays, airway surface liquid and particle-tracking microrheology, for our endocervical culture system. Endocervical cells treated with CFTR inhibitors demonstrated dehydrated, thicker mucus secretions compared to controls in both assay outputs. Our studies suggest that CFTR may be an important mediator of fertility changes and provide experimental evidence for the infertility phenotype seen in women with cystic fibrosis. Additionally, assays developed in these studies provide new endpoints for assessing cervical mucus changes in vitro.

Vitrification is increasingly used to cryopreserve gametes and embryos in assisted reproductive technology. Our prior research demonstrates that vitrification preserves the viability and functionality of follicles. However, its impact on oocyte remains unknown. The current study investigates whether vitrification maintains the oocyte transcriptome during in vitro follicle development and oocyte maturation. Immature mouse preantral follicles were vitrified, then warmed and cultured in vitro for 8 days to grow to the preovulatory stage, followed with induction of ovulation and oocyte maturation on day 9, with fresh follicles as the control. Oocytes at germinal vesicle (GV) stage from grown preovulatory follicles on day 8 and oocytes at metaphase II (MII) post-ovulation on day 9 were collected for single-oocyte Smart-Seq2 RNA sequencing. Principal component analysis separated GV and MII oocytes into two distinct clusters, but oocytes from fresh and vitrified follicles largely overlapped. Differentially expressed genes (DEG) analysis revealed that oocytes from fresh and vitrified follicles, at either GV or MII stage, had comparable expression of maternal effect genes and other genes related to oocyte meiotic and developmental competence. There was a significant transcriptomic change in oocytes during GV-to-MII transition. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified DEGs between GV and MII oocytes related to cell cycle, RNA processing, mitochondrion, and ribosome. In summary, our study demonstrates that vitrification preserves oocyte transcriptome during in vitro follicle development and oocyte maturation, supporting its potential for fertility preservation. Moreover, key DEGs identified during GV-to-MII transition indicate their potential functions in oocyte meiotic and developmental competence.

Benign recurrent intrahepatic cholestasis (BRIC) is a rare genetic liver disorder characterized by recurrent episodes of jaundice and severe pruritus without significant liver damage. Here, we present the case of a man in his early 20s, standing 143 cm tall, who has experienced recurrent jaundice and pruritus over an 18-year period. Laboratory investigations and liver biopsy examination indicated typical intrahepatic cholestasis with normal gamma-glutamyl transpeptidase levels, excluding viral, metabolic, and autoimmune causes. Further genetic analysis confirmed the diagnosis of BRIC with two compound heterozygous mutations in ATP8B1 gene on chromosome 18: NM_001374385.1: c.2081T > C p. (Ile694Thr) (rs541474497) in exon 18 and NC_000018.10 (NM_001374385.1): c.1631-2A > G in intron 15. To our knowledge, the latter has not been reported. Family lineage analysis and Sanger sequencing showed that these mutations were de novo rather than hereditary. Treatment with ursodeoxycholic acid and cholestyramine resulted in complete resolution of jaundice and pruritus, with normalization of serum bilirubin level after six months of therapy.

Encephalomyocarditis virus (EMCV) causes sporadic and epizootic outbreaks among various domesticated and non-domesticated animal species worldwide. Although outbreaks are mostly reported in domestic pigs, mortality is reported in elephants, ungulates, nonhuman primates (NHPs), and rodents. Rats of the genus Rattus serve as primary reservoirs and vectors, but alternative infection routes have been proposed. Clinical disease is characterized by acute heart failure in most taxonomic groups, often culminating in rapid death. Due to the rapid progression of the disease, diagnostic confirmation is most commonly obtained postmortem. Pathological examination reveals interstitial lymphohistiocytic myocarditis and multiorgan congestion in most cases. EMCV is often demonstrated with RT-PCR or virus isolation techniques, but other methods, e.g., serology and immunohistochemistry, are available. The rapid progression of EMCV precludes effective therapeutic intervention, though agents such as interferon, verapamil, and curcumol have shown potential efficacy. Preventative strategies are crucial, emphasizing biosecurity measures to mitigate rodent contamination of feed and water. Inactivated vaccines have demonstrated protective efficacy in experimental models involving mice, pigs, and elephants, with analogous immunogenic responses observed in various zoological species. Live attenuated vaccines have conferred protection in pigs and NHPs, albeit with variable seroconversion rates in different species.

The success of chimeric antigen receptor (CAR)-T cell (Tc) immunotherapy in refractory B-cell acute lymphoblastic leukemia (B-ALL) suggests adaptation of this strategy toward HIV. Because cytomegalovirus (CMV) vaccine vectors generated Tc responses that controlled viral replication, these studies aim to genetically modify CMV-specific Tc with HIV-CAR2 vectors and link HIV immunotherapy to persistent CMV antigen stimulation. To mimic a clinical scenario, rhesus macaques were challenged with the CCR5-tropic simian/human immunodeficiency virus (SHIV-D) prior to antiretroviral therapy (ART). Autologous CMV-specific Tc were transduced with the control CEA-CAR2 or CD4-CAR2/maC46 vectors and reinfused. After stopping ART, the plasma viral load (PVL) in the control rebounded and was sustained above 1.7 × 10 ⁴ copies/mL; PVL in CD4-CAR2-treated animals was delayed up to 6 weeks and 10-fold lower. The CD4 CAR-Tc frequency peaked at day 7 and was detected in lymphoid tissues at 6 weeks. Both CEA-CAR2 and CD4-CAR2 persisted in PBMCs for about 2 years, which indicates that the CMV-specific CAR Tc were maintained based on their CMV specificity. However, long-term PVL was stable in all animals. Thus, CMV-specific CAR-Tc were active initially, persisted long term, but failed to control viral replication. IMPORTANCE Because of latent viral reservoirs and a dysfunctional immune response, HIV replication rebounds when antiretroviral therapy is interrupted. Therefore, cytomegalovirus (CMV)-specific Tc were genetically modified with anti-HIV CD4-CAR2 vectors to link the targeting of the HIV envelope to the persistent CMV immune response. In this clinical scenario with simian/human immunodeficiency virus (SHIV) challenge and antiretroviral therapy (ART) suppression, early activity of the CAR Tc delayed rebound in the rhesus macaque/SHIV challenge model. However, even with long-term persistence of CAR Tc in the blood, control of viral replication was not achieved. These data suggest that CAR Tc will require additional interventions to cure HIV infection.

Background Wound healing is a complex physiological process essential for restoring tissue integrity following various injuries, ranging from minor, everyday incidents to post-surgical complications. Emerging studies have demonstrated that lactic acid bacteria (LAB) can offer benefits beyond gut health, extending their positive effects on skin health. This study investigated the potential of Lactobacillus reuteri NCHBL-005, a honeybee-derived probiotic strain, to enhance fibroblast-mediated wound healing. Method L929 cells and mouse embryonic fibroblasts (MEFs) were utilized as models to specifically target fibroblasts. To assess the wound healing potential in vitro, a scratch assay was performed, providing insights into wound closure. Additionally, we created wound models in mice to evaluate the in vivo effects of the treatment. Results Our results showed that L. reuteri NCHBL-005 significantly accelerated wound closure in L929 fibroblast compared to other lactobacilli and exhibited superior efficacy in activating the mitogen-activated protein kinase (MAPK) pathway. Through MAPK inhibition assays, we confirmed that the wound healing effects of L. reuteri NCHBL-005 were MAPK-dependent, promoting fibroblast proliferation and differentiation. Notably, L. reuteri NCHBL-005 treatment did not facilitate wound healing in MEF cells derived from Toll-like-receptor 2 knockout (TLR2 −/− ) mice, highlighting the critical role of TLR2 in this mechanism. In vivo studies further corroborated these findings, in which topical administration of L. reuteri NCHBL-005 enhanced wound healing and stimulated fibroblast proliferation and activation, as confirmed by histopathological analysis. Conclusion These findings revealed that L. reuteri NCHBL-005 activates fibroblasts through TLR2 stimulation and subsequent MAPK pathway activation, suggesting its potential as a promising therapeutic candidate for wound management.

The most dynamic and repetitive regions of great ape genomes have traditionally been excluded from comparative studies1, 2–3. Consequently, our understanding of the evolution of our species is incomplete. Here we present haplotype-resolved reference genomes and comparative analyses of six ape species: chimpanzee, bonobo, gorilla, Bornean orangutan, Sumatran orangutan and siamang. We achieve chromosome-level contiguity with substantial sequence accuracy (<1 error in 2.7 megabases) and completely sequence 215 gapless chromosomes telomere-to-telomere. We resolve challenging regions, such as the major histocompatibility complex and immunoglobulin loci, to provide in-depth evolutionary insights. Comparative analyses enabled investigations of the evolution and diversity of regions previously uncharacterized or incompletely studied without bias from mapping to the human reference genome. Such regions include newly minted gene families in lineage-specific segmental duplications, centromeric DNA, acrocentric chromosomes and subterminal heterochromatin. This resource serves as a comprehensive baseline for future evolutionary studies of humans and our closest living ape relatives.

Major developmental events occurring in the hippocampus during the third trimester of human gestation and neonatally in altricial rodents include rapid and synchronized dendritic arborization and astrocyte proliferation and maturation. We tested the hypothesis that signals sent by developing astrocytes to developing neurons modulate dendritic development in vivo. First, we altered neuronal development by exposing neonatal (third trimester‐equivalent) mice to ethanol, which increased dendritic arborization in hippocampal pyramidal neurons. We next assessed concurrent changes in the mouse astrocyte translatome by translating ribosomal affinity purification (TRAP)‐seq. We followed up on ethanol‐inhibition of astrocyte Chpf2 and Chsy1 gene translation because these genes encode biosynthetic enzymes of chondroitin sulfate glycosaminoglycan (CS‐GAG) chains (extracellular matrix components that inhibit neuronal development and plasticity) and have not been explored before for their roles in dendritic arborization. We report that Chpf2 and Chsy1 are enriched in astrocytes, and their translation is inhibited by ethanol, which also reduces the levels of CS‐GAGs measured by Liquid Chromatography/Mass Spectrometry. Finally, astrocyte‐conditioned medium derived from Chfp2 ‐silenced astrocytes increased neurite length and branching of hippocampal neurons in vitro, mechanistically linking changes in CS‐GAG biosynthetic enzymes in astrocytes to altered neuronal development. These results demonstrate that CS‐GAG biosynthetic enzymes in astrocytes regulate dendritic arborization in developing neurons and are involved in ethanol‐induced altered neuronal development.

The effectiveness of COVID-19 mRNA vaccines is diminished in organ transplant patients. Using a multi-omics approach, we investigate the immunological state of lung transplant (LTX) recipients at baseline and after SARS-CoV-2 mRNA vaccination compared to healthy controls (HCs). LTX patients exhibit a baseline immune profile resembling severe COVID-19 and sepsis, characterized by elevated pro-inflammatory cytokines (e.g., EN-RAGE [also known as S100A12], interleukin [IL]-6), reduced human leukocyte antigen (HLA)-DR expression on monocytes and dendritic cells, impaired cytokine production, and increased plasma microbial products. Single-cell RNA sequencing identifies an enriched monocyte cluster in LTX patients marked by high S100A family expression and reduced cytokine and antigen presentation genes. Post vaccination, LTX patients show diminished antibody, B cell, and T cell responses, along with blunted innate immune signatures. Integrative analysis links these altered baseline immunological features to impaired vaccine responses. These findings provide critical insights into the immunosuppressed condition of LTX recipients and their reduced vaccine-induced adaptive and innate immune responses.

Nonhuman primates with long-term cranial implant chambers require regular chamber cleaning with antiseptic solutions. The purpose of this study was to determine if ATP bioluminescence testing can be used to identify microbial contamination of fomites and environmental samples in the context of cranial implant chamber cleaning procedures. ATP bioluminescent swab samples were compared with traditional bacterial culture swab samples from the same sources collected during the scheduled chamber maintenance procedures for Rhesus macaques ( Macaca mulatta ) that are part of studies related to the modulation of brain circuits in parkinsonism. Over the course of 17 d, samples were collected from the chamber rims, forceps (pre- and postcleaning), povidone-iodine bottle, quaternary ammonium and alcohol-based disinfectant solution containers, and cotton ball jar (ATP swab: n = 10 per environmental source; bacterial culture swab: n = 6 per environmental source). Chamber rims yielded the highest ATP relative light unit values compared with the other environmental sample groups and heavy growth on bacterial culture. A total of 16/36 (44%) swab samples from environmental sources yielded growth on bacterial culture, and clinically relevant bacterial species were identified in samples from the chamber rims, cotton ball jar, povidone-iodine bottle, and forceps. Although high ATP RLU levels and positive bacterial growth were identified for these environmental samples, there was a poor correlation between the ATP RLU values with the semiquantitative bacterial culture scores. Based on the results of this study, a high ATP RLU cutoff threshold would be needed to maximize the accuracy of using this method instead of bacterial culture to identify potential sources of microbial contamination. This study represents the first published microbial surveillance investigation of environmental samples from materials and solutions used in cranial implant chamber maintenance.

Chronic psychosocial stress is associated with increased risk of psychiatric disorders. Magnetic resonance spectroscopy (MRS) in humans has been used to show that glutamate levels in medial prefrontal cortex (mPFC) following acute stress exposure adapt to recent chronic stress levels. Here, we sought to determine the presence of this glutamate stress response adaptation in rhesus macaques, whose societies are maintained by dominance relationships that are enforced by agonistic interactions and result in chronic stress phenotypes seen in humans. We tested the hypothesis that change in mPFC glutamate after an acute stressor would be moderated by behavioral factors related to social subordination in a manner similar to that previously observed in humans. Seventeen adult female rhesus monkeys (Macaca mulatta, 13–23 yrs.) were observed over ten weeks to collect behavioral data and then received two MRS scans. The first scan occurred after acute stress manipulation involving relocation and isolation. The second control scan occurred after acclimation to the new location. As expected, we found that a behavioral measure of social subordination predicted an adaptive glutamate response such that animals experiencing more submissive behavior asymmetry (a behavioral measure related to social subordination) exhibited an attenuated glutamate response to the acute stressor. These data establish the use of MRS to measure the adaptive glutamate stress in non-human primates and will help further our understanding of the neurobiology of stress adaptation.

Initial interactions between HIV-1 and the immune system at mucosal exposure sites play a critical role in determining whether the virus is eliminated or progresses to establish systemic infection. The virus that successfully crosses the mucosal barrier to establish infection in the new host is referred to as the transmitted/founder (TF) virus. Following mucosal HIV-1 transmission, type 1 interferons (IFN-I) are rapidly induced at sites of initial virus replication. The resistance of TF variants to these antiviral effects of the IFN-I has been studied among HIV-1 subtypes B and C. However, their role in restricting HIV-1 replication among subtypes D and AD recombinant remains unexplored. This study assessed the sensitivity of HIV-1 subtype D and AD recombinant TF viruses to IFN-I by infecting peripheral blood mononuclear cells in vitro with infectious molecular clones of these viruses. Cells were exposed to varying concentrations of interferon-α and interferon-β, and viral replicative capacity was measured using HIV-1 p24 antigen ELISA from culture supernatants. Sensitivity to IFN-I was quantified based on viral replication levels. The results showed that interferon-α was more effective in inhibiting viral replication than interferon-β, regardless of the varying amounts of IFN-I used. However, recombinant AD viruses were found to be more resistant to the antiviral effects of IFN-I compared to subtype D viruses. These findings highlight the differential sensitivity of HIV-1 subtypes AD recombinant and D TF viruses to IFN-I and underscore the potential of IFN-I as a therapeutic strategy to target TF viruses and reduce HIV-1 transmission, particularly in populations where subtype D is prevalent.

Folliculogenesis encompasses many stages as the somatic granulosa and theca cells support oocytes through growth and maturation. A novel follicle stage, between primordial and transitional stages, was identified in mice and defined as “zip”. Like all other follicle stages, the “zip” stage is characterized by its granulosa cell morphology. The “wedge” GC morphology in zip follicles is predicted to be the first granulosa cell division, marking the transition from squamous to cuboidal morphology. Here, zip and transitional stages were identified in histological sections of porcine, bovine, rhesus monkey, and human ovaries. Several growth dynamics characterized at these follicle stages were conserved between species. Oocyte diameter and area increased between the primordial and transitional stages in the porcine ovary and between the primordial and primary stages in the rhesus monkey ovary but appeared unchanged in bovine and human ovaries. In all species except for pigs, granulosa cell number and height increased at stages earlier than observed changes in the oocyte. Furthermore, there were differences in the percentage of zip and transitional follicle stages present in the cortical region across species. This implies that there may be species-dependent activation and growth mechanisms that require further study. The parameters defined here for identifying and characterizing the zip and transitional follicle stages across species can act as a tool for measuring factors that perturb or induce primordial follicle activation or effect follicle morphometric parameters in support of future innovations for fertility preservation and restoration.

We assess the proposition that intergroup conflict (IGC) in non-human primates offers a useful comparison for studies of human IGC and its links to parochial altruism and prosociality. That is, for non-linguistic animals, social network integration and maternal influence promote juvenile engagement in IGC and can serve as the initial grounding for sociocultural processes that drive human cooperation. Using longitudinal data from three cohorts of non-adult vervet monkeys ( Chlorocebus pygerythrus ), we show that non-adults are sensitive to personal (age) and situational risk (participant numbers). The frequency and intensity of participation, although modulated by rank and temperament, both mirrors maternal participation and reflects non-adult centrality in the grooming network. The possibility of social induction is corroborated by the distribution of grooming during IGC, with non-adults being more likely to be groomed if they were female, higher-ranking and participants themselves. Mothers were more likely to groom younger offspring participants of either sex, whereas other adults targeted higher-ranking female participants. Although we caution against a facile alignment of these outcomes to human culturally mediated induction, there is merit in considering how the embodied act of participation and the resultant social give-and-take might serve as the basis for a unified comparative investigation of prosociality.