Microglial-derived C1q integrates into neuronal ribonucleoprotein complexes and impacts protein homeostasis in the aging brain.

Neuroimmune interactions mediate intercellular communication and underlie critical brain functions. Microglia, CNS-resident macrophages, modulate the brain through direct physical interactions and the secretion of molecules. One such secreted factor, the complement protein C1q, contributes to complement-mediated synapse elimination in both developmental and disease models, yet brain C1q protein levels increase significantly throughout aging. Here, we report that C1q interacts with neuronal ribonucleoprotein (RNP) complexes in an age-dependent manner. Purified C1q protein undergoes RNA-dependent liquid-liquid phase separation (LLPS) in vitro, and the interaction of C1q with neuronal RNP complexes in vivo is dependent on RNA and endocytosis. Mice lacking C1q have age-specific alterations in neuronal protein synthesis in vivo and impaired fear memory extinction. Together, our findings reveal a biophysical property of C1q that underlies RNA- and age-dependent neuronal interactions and demonstrate a role of C1q in critical intracellular neuronal processes.

CXCR2 inhibition in G-MDSCs enhances CD47 blockade for melanoma tumor cell clearance.

The use of colony-stimulating factor-1 receptor (CSF1R) inhibitors has been widely explored as a strategy for cancer immunotherapy due to their robust depletion of tumor-associated macrophages (TAMs). While CSF1R blockade effectively eliminates TAMs from the solid tumor microenvironment, its clinical efficacy is limited. Here, we use an inducible CSF1R knockout model to investigate the persistence of tumor progression in the absence of TAMs. We find increased frequencies of granulocytic myeloid-derived suppressor cells (G-MDSCs) in the bone marrow, throughout circulation, and in the tumor following CSF1R deletion and loss of TAMs. We find that G-MDSCs are capable of suppressing macrophage phagocytosis, and the elimination of G-MDSCs through CXCR2 inhibition increases macrophage capacity for tumor cell clearance. Further, we find that combination therapy of CXCR2 inhibition and CD47 blockade synergize to elicit a significant anti-tumor response. These findings reveal G-MDSCs as key drivers of tumor immunosuppression and demonstrate their inhibition as a potent strategy to increase macrophage phagocytosis and enhance the anti-tumor efficacy of CD47 blockade in B16-F10 melanoma.

Neogenin-1 distinguishes between myeloid-biased and balanced Hoxb5+ mouse long-term hematopoietic stem cells.

Hematopoietic stem cells (HSCs) self-renew and generate all blood cells. Recent studies with single cell transplants and lineage tracing suggest that adult HSCs are diverse in their reconstitution and lineage potentials. However, prospective isolation of these subpopulations has remained challenging. Here, we identify Neogenin-1 (NEO1) as a unique surface marker on a fraction of mouse HSCs labeled with Hoxb5, a specific reporter of long-term HSCs (LT-HSCs). We show that NEO1+Hoxb5+ LT-HSCs expand with age and respond to myeloablative stress in young mice while NEO1-Hoxb5+ LT-HSCs exhibit no significant change in number. Furthermore, NEO1+Hoxb5+ LT-HSCs are more often in the G2/S cell cycle phase compared to NEO1-Hoxb5+ LT-HSCs in both young and old bone marrow. Upon serial transplantation, NEO1+Hoxb5+ LT-HSCs exhibit myeloid-biased differentiation and reduced reconstitution while NEO1-Hoxb5+ LT-HSCs are lineage-balanced and stably reconstitute recipients. Gene expression analysis reveals erythroid and myeloid priming in the NEO1+ fraction and association of quiescence and self-renewal-related transcription factors with NEO1- LT-HSCs. Finally, transplanted NEO1+Hoxb5+ LT-HSCs rarely generate NEO1-Hoxb5+ LT-HSCs while NEO1-Hoxb5+ LT-HSCs repopulate both LT-HSC fractions. This supports a model in which dormant, balanced NEO1-Hoxb5+ LT-HSCs can hierarchically precede active, myeloid-biased NEO1+Hoxb5+ LT-HSCs.

Mechanism of substrate specificity in 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidases.

5'-Methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidase (MTAN) plays a key role in the methionine-recycling pathway of bacteria and plants. Despite extensive structural and biochemical studies, the molecular mechanism of substrate specificity for MTAN remains an outstanding question. Bacterial MTANs show comparable efficiency in hydrolyzing MTA and SAH, while the plant enzymes select preferentially for MTA, with either no or significantly reduced activity towards SAH. Bacterial and plant MTANs show significant conservation in the overall structure, and the adenine- and ribose-binding sites. The observation of a more constricted 5'-alkylthio binding site in Arabidopsis thalianaAtMTAN1 and AtMTAN2, two plant MTAN homologues, led to the hypothesis that steric hindrance may play a role in substrate selection in plant MTANs. We show using isothermal titration calorimetry that SAH binds to both Escherichia coli MTAN (EcMTAN) and AtMTAN1 with comparable micromolar affinity. To understand why AtMTAN1 can bind but not hydrolyze SAH, we determined the structure of the protein-SAH complex at 2.2Å resolution. The lack of catalytic activity appears to be related to the enzyme's inability to bind the substrate in a catalytically competent manner. The role of dynamics in substrate selection was also examined by probing the amide proton exchange rates of EcMTAN and AtMTAN1 via deuterium-hydrogen exchange coupled mass spectrometry. These results correlate with the B factors of available structures and the thermodynamic parameters associated with substrate binding, and suggest a higher level of conformational flexibility in the active site of EcMTAN. Our results implicate dynamics as an important factor in substrate selection in MTAN.

Proteomic analysis of young and old mouse hematopoietic stem cells and their progenitors reveals post-transcriptional regulation in stem cells.

The balance of hematopoietic stem cell (HSC) self-renewal and differentiation is critical for a healthy blood supply; imbalances underlie hematological diseases. The importance of HSCs and their progenitors have led to their extensive characterization at genomic and transcriptomic levels. However, the proteomics of hematopoiesis remains incompletely understood. Here we report a proteomics resource from mass spectrometry of mouse young adult and old adult mouse HSCs, multipotent progenitors and oligopotent progenitors; 12 cell types in total. We validated differential protein levels, including confirmation that Dnmt3a protein levels are undetected in young adult mouse HSCs until forced into cycle. Additionally, through integrating proteomics and RNA-sequencing datasets, we identified a subset of genes with apparent post-transcriptional repression in young adult mouse HSCs. In summary, we report proteomic coverage of young and old mouse HSCs and progenitors, with broader implications for understanding mechanisms for stem cell maintenance, niche interactions and fate determination.

Esophageal Stricture Caused by Actinomyces in a Patient with No Apparent Predisposing Factors.

Actinomyces species are Gram positive anaerobic or microaerophilic bacteria that are part of the human flora in the oropharyngeal, gastrointestinal, and genitourinary tract. In the presence of a mucosal injury, they can become pathogenic and infect the underlying tissue without respect for tissue planes, leading to abscesses, fistulas, and sinus tracts. Through contiguous and hematogenous spread, virtually any organ can become infected. The presentation can be myriad and often mimics tumors, tuberculosis, or other more common infections and inflammatory conditions. While the cervicofacial region is the most common site of infection, involvement of the esophagus is unusual. Esophageal actinomycosis mostly occurs in patients with compromised immunity or prior esophageal injuries. Occurrence in immunocompetent individuals without risk factors is exceedingly rare. We report a case of esophageal actinomycosis complicated by an esophageal stricture in a patient with no apparent predisposing conditions.

A case of esophagogastroduodenoscopy induced Takotsubo cardiomyopathy with complete heart block.

Takotsubo cardiomyopathy (TC) describes a reversible left ventricular dysfunction characterized by apical ballooning and basal hyperkinesis, commonly triggered by emotional or physical distress. TC associated with an esophagogastroduodenoscopy (EGD) has rarely been reported. We report a case of TC with complete heart block (CHB) in a patient receiving an EGD, who had no underlying cardiac disease, had previously tolerated both local and general anesthesia, and who had previously undergone similar endoscopic procedures without complications. The concurrence of both TC and CHB is unique in this case pertaining to a patient with no significant risk factors. The incidence, mechanism and prognosis of TC-associated arrhythmias are also reviewed.

A delayed presentation of splenic laceration and hemoperitoneum following an elective colonoscopy: A rare complication with uncertain risk factors.

Splenic laceration is a rare yet often underreported complication of colonoscopy that is infrequently discussed with the patient during the consent process. Most cases present within 48 h after the inciting colonoscopy; a delayed presentation is rare. We present a case of splenic laceration with hemoperitoneum that manifested 5 days after the initial colonoscopy. The patient was treated conservatively. Traditionally perceived risk factors such as intra-abdominal adhesions, splenomegaly, anticoagulation use, biopsy, polypectomy, a technically challenging procedure, and anesthesia assistance have not been clearly shown to increase the incidence of splenic injury following a colonoscopy. Since the risk factors of splenic injury remain unclear, the clinical presentation is nonspecific, and the consequences can be serious, the endoscopist should make an effort to inform the patient of this rare complication before the procedure.

The neural correlates of performance in adolescents at risk for schizophrenia: inefficiently increased cortico-striatal responses measured with fMRI.

BACKGROUND: fMRI studies indicate that schizophrenia patients and their adult relatives require greater prefrontal activation to maintain performance at levels equal to controls, but studies have not established if this pattern of inefficiency is observed in child and adolescent offspring of schizophrenia patients (SCZ-Off). METHODS: Using a task with visual working memory demands, we investigated activation in cortico-striatal networks and dorsal prefrontal modulation of regions underlying visual working memory in a group of SCZ-Off (n = 19) and controls with no family history of psychosis (n = 25 subjects) using an event-related design. Trials were divided based on memory performance (correct vs. incorrect) to specifically identify the neural correlates of correct working memory performance. RESULTS: Whereas groups did not differ in terms of behavioral accuracy, SCZ-Off demonstrated significantly increased fMRI-measured activation in dorsal prefrontal cortex and the caudate nucleus during correct, relative to incorrect memory performance. Whereas activation in SCZ-Off was high and independent of performance in each region, in controls the fMRI response was related to behavioral proficiency in the caudate. Further, exploratory analyses indicated that this inefficiency in the dorsal prefrontal cortex response increased with age in SCZ-Off (but in no other regions or group). Finally, these differences were not based in differences in dorsal prefrontal modulation of other regions during successful performance. DISCUSSION: These results are consistent with observed patterns in adult patients and first-degree relatives. Inefficient fronto-striatal responses during working memory may characterize the schizophrenia diathesis and may reflect the effects of the illness and vulnerability for the illness.