Background
Over 500 clinical trials combining radiation (RT) and immune checkpoint blockade (ICB) have been initiated based on preclinical evidence that RT can augment local immunity and improve the efficacy of ICB. However, the majority of clinical trials have found no benefit of combining RT and ICB, raising questions about whether a synergy exists. We examined whether RT and ICB interact to beneficially stimulate the immune response in patients and explored genomic biomarkers of response to combination RT and ICB.

Methods
We analyzed 2,046 patients from 4 cohorts. The COSINR dataset is a randomized clinical trial of 22 patients with non-small cell lung cancer (NSCLC) treated with concurrent or sequential RT and ICB. Paired pre/on-treatment biopsies of an irradiated metastasis underwent exome and RNA-seq. On-treatment biopsies were obtained after RT and prior to ICB (sequential) or after RT and one cycle of ICB (concurrent). The UC cohort contains targeted sequencing of 58 NSCLCs treated with ICB alone, sequential RT and ICB, or concurrent RT and ICB. The SU2C cohort comprises targeted sequencing of 306 ICB-treated NSCLCs from 11 institutions. The pan-cancer MSKCC cohort contains targeted sequencing of 1,660 patients treated with ICB. 

Aneuploidy score (AS) was defined as the fraction of chromosome arms with arm-level copy number alterations. Single-sample gene set enrichment analysis was used to compute Hallmark immune signaling pathway scores. Survival analyses utilized the Kaplan-Meier method and Cox proportional hazards models. 

Results
In the COSINR trial, RT and ICB increased, whereas RT alone decreased, expression of effector T-cell IFN-gamma and adaptive immune signatures (P<0.05). Baseline AS was negatively associated with expression of interferon, interleukin, and inflammatory signaling pathways (P<0.05). High AS was independently associated with poor OS following ICB after controlling for TMB and ICB paradigm in the MSKCC cohort (hazard ratio[95% CI]: 1.44[1.07–1.94], P=0.014) and controlling for TMB in the SU2C cohort (hazard ratio[95% CI]: 3.06[1.58–5.94], P=9.5e-4). In the UC cohort, high (≥median) AS tumors treated with concurrent RT and ICB had superior 1-year OS vs. sequential RT and ICB or ICB alone (76% vs. 38% vs. 31%, P=0.044). RT did not improve OS in low AS patients. Our findings were corroborated in the COSINR cohort: patients whose tumors exhibited high, but not low, AS had improved survival when treated with concurrent vs. sequential RT and ICB (1-year overall survival [OS] 100% vs. 17%, P=0.025).

Conclusions
Our findings distinguish the genomic and transcriptomic effects of RT versus RT and ICB and challenge the prevailing paradigm that local ablative RT alone positively stimulates the immune response. We propose the use of aneuploidy as a biomarker in personalizing immunotherapeutic treatment approaches. Concurrent RT and ICB may augment survival in patients with highly aneuploid, immunologically cold tumors.

Concurrent radiation and immunotherapy augments immunity and survival in aneuploid lung cancer

Introduction
Colon cancer is one of the top five leading causes of cancer in both males and females. There is a well-known association between colon cancer and microbial infections. The presence of certain bacterial infections in the appropriate clinical context should prompt further investigation of colon cancer.

Case Presentation
A 41-year-old male with history of alcohol use disorder and bipolar disorder who presented for acute encephalopathy after cardiac arrest with return of spontaneous circulation within 2 minutes in the field.
In the emergency department he was found to be hypotensive and hypoxic; labs revealed leukocytosis (50.9 K/ul) with bandemia (15%), anemia (5.9g/dl), and LFTs consistent with acute liver failure. CT abdomen/pelvis showed hepatosplenomegaly, pelvic ascites, colonic wall thickening and luminal narrowing in mid-sigmoid colon.
He was admitted to the ICU for twelve days for septic shock intermittently requiring vasopressors, acute liver failure, and respiratory failure. A diagnostic paracentesis revealed a transudative effusion; blood cultures were positive for Clostridium septicum and Enterococcus avium.
A subsequent colonoscopy demonstrated a sigmoid colonic mass whose pathology was consistent with adenocarcinoma. Due to his acute decompensated liver failure secondary to alcohol use; he was not a candidate for surgery. The family placed the patient in hospice care and he eventually expired.

Discussion
Colon malignancy has been associated to certain bacterial strains, with the most common being Streptococcus bovis/gallolyticus, and less frequently Clostridium septicum, Enterococci species, Bacteroides fragilis, Fusobacterium nucleatum, and other Streptococci species. In fact, of the approximately 106,180 new cases of colon cancer reported yearly, 10-25% are associated with infections caused by Streptococcus bovis and Clostridium septicum.
Clostridium septicum is an opportunistic, gram-positive anaerobe that has several virulence factors, including alpha toxin. It can survive in acidic environments due to its ability to produce spores and has the capability to perforate the colonic epithelial lining, releasing spores into the blood and leading to bacteremia. The risk factors for C. septicum infection include immunosuppressed states, hematological malignancies, neutropenia, and diabetes mellitus. Unfortunately, C. septicum infection has a high mortality rate of 60%, which can only be diagnosed through positive cultures.
Treatment for C. Septicum infection varies depending on the presentation, as it can cause aortitis, myonecrosis, gangrene, cellulitis, septic shock, fasciitis, hemolysis, and abscess. However, early antibiotics and possible surgical intervention are recommended for effective management. Piperacillin/tazobactam or penicillin G are the first-line treatments for C. septicum infection. Therefore, it is crucial to identify and manage this infection as early as possible to prevent the high mortality rate associated with it.
Conclusion
Clostridium septicum infection and colonic malignancy is a well-known association therefore clinicians should have a high suspicion for colon cancer in patients who present with this infection.

A Potentially Lethal Harbinger: Clostridium Septicum Bacteremia in a Patient Found to have Adenocarcinoma of the Colon

Background: Acinetobacter baumannii is one of the world’s most multi-drug resistant (MDR) pathogens. Traditionally responsible for causing nosocomial pneumonia, bacteremia, and soft tissue infections in critically ill and immunocompromised patients, A. baumannii has evolved to propagate a growing number of atypical infections, including fatal cases of necrotizing fasciitis (NF; i.e., flesh-eating disease), over the past decade. Concerningly, these NF A. baumannii (NFAb) isolates display enhanced virulence and antibiotic resistance uncharacteristic of type strains, resulting in limited efficacy of current therapeutics. As such, understanding of mechanisms by which these strains evade host defenses, environmental stress, and previously effective antibiotics is imperative, yet remains scant. Our lab has obtained two MDR NFAb isolates from a case of NF that led to sepsis and death over 36-hours. The present study sought to elucidate resistance mechanisms of these isolates for the identification of new drug targets to treat MDR A. baumannii infections.

Methods: To determine if NFAb resistance and propagation of infection involves the mechanism of host immune cell entry, NFAb isolates (i.e., NFAb-1 collected pre-mortem from blood and NFAb-2 collected post-mortem from tissue) were co-incubated with human-derived THP-1 macrophages with or without pre-treatment with Cytochalasin D, an inhibitor of phagocytosis. A colistin-protection assay was performed to isolate viable intracellular bacteria, which were quantified by colony forming units (CFUs) after 24 hours. To assess resistance to desiccation, dried NFAb isolates and type strains were resuspended, and surviving cells were quantified by CFUs at 1, 5, and 7 day intervals. Lastly, transformation efficiency was assessed using selective media and fluorescence via flow cytometry, then reported as transformant CFU:total CFU.

Results: Similar to type strains, NFAb isolates were found to primarily enter macrophages via macrophage-mediated phagocytosis; however, both NFAb isolates displayed greater intracellular entry and survival in the host immune cells than type strains. Interestingly, NFAb-1 demonstrated greater resistance to desiccation than NFAb-2 over the course of 7 days. Although previous studies highlighted genomic differences between NFAb-1 and NFAb-2, no differences were observed in transformation efficiency between NFAb isolates and type strains.

Conclusion: Collectively, these results suggest NFAb isolates utilize a variety of resistance strategies to survive, and possess a profound ability to rapidly adapt, even within the context of a single infection. Moving forward, we aim to identify the genes propagating the intracellular survival and infection demonstrated by these isolates to serve as potential targets for novel therapeutics.

Utilizing Atypical Isolates of Acinetobacter baumannii to Uncover Novel Mechanisms of Resistance

The role of Lcn2 in Triple Negative Breast Cancer Organoids

Background: Research shows that increased severity of congenital heart disease (CHD) is associated with increased frequency of neurological deficits in children with structural cardiac abnormalities. Young children with more complex CHDs who require cardiopulmonary bypass (CPB) have a significantly higher incidence of developmental abnormalities and neurological deficits, including academic difficulties and behavioral abnormalities. Microglia, the resident immune cells in the central nervous system, play a key role in neural immune function. Mesenchymal stromal cells (MSCs) are multipotent, non-hematopoietic cells that possess immunomodulatory and regenerative properties and regulate microglia activation. It is hypothesized that intra-arterial MSC delivery through CPB in a survival porcine model is neuroprotective by modulating systemic and brain-specific inflammatory responses.

Methods: To mimic the effects in young CHD patients, two-week old female piglets were randomly assigned to one of three treatment groups: control, CPB, and CPB+MSC. The CPB+MSC group consisted of a CPB surgery followed by MSC administration before weaning. Microglia undergo morphological changes from a naïve, ramified to an amoeboid shape under activated condition. To analyze the effects of MSCs on microglia activation, changes in the length of microglial processes and number of branch points from the nuclei were assessed.

Results: Four weeks post-surgery, analysis using Imaris software showed that CPB reduced both the length of processes and number of branch points compared to control in three different cortex regions. CPB+MSC revealed significantly longer processes (p=0.0281) and more branch points (p=0.0232) than CPB alone, indicating that morphological changes in microglia were normalized after MSC delivery.

Conclusion: MSCs regulate microglial activation and participate in the phenotypic switch from a pro-inflammatory state to a repair-permissive state. In contrast to their ramified morphology under normal conditions, activated microglia undergo structural remodeling and adopt an amoeboid morphology with highly retracted processes, a hallmark of brain inflammation, as shown in the CPB group. Microglia routinely migrate toward sites of neuronal degeneration induced by excitotoxic injury. In MSC-treated animals, there are significantly fewer amoeboid microglia with an associated increase in the length of processes and number of branch points to mitigate the effects of CPB, indicating attenuated activation of the microglia. MSC delivery during CPB is highly effective and shows translational potential to minimize CPB-induced systemic inflammation and microglial activation in children with CHD.

Effect of mesenchymal stromal cell delivery through cardiopulmonary bypass in a piglet model

Background: 
Monkeypox, caused by the Monkeypox virus (MPV), is a double-stranded DNA virus belonging to the genus Orthopoxvirus. While confirmed cases of Mpox disease have been reported in since the 1970s, the recent global outbreak has escalated Monkeypox to a public health emergency. Although Monkeypox is typically considered a self-limiting disease, the recent outbreak has demonstrated an increase in disease morbidity and mortality, particularly in those with immunocompromised status. For those with HIV infection, co-infection with Monkeypox can lead to atypical disease presentation. This poster highlights an example of a highly atypical dermatological presentation in a patient with HIV and Monkeypox co-infection. 

Discussion: A 28-year-old male with a history of untreated HIV/AIDS presented with diffuse bilateral eschars with well-extended surrounding erythema for two days. The lesions were expansive and located diffusely across the bilateral upper extremities. The patient reported the lesions were slowly enlarging and mildly pruritic. Additionally, bilateral tender inguinal lymphadenopathy and a single painful shallow ulcer on the right-side penile shaft was present. While Monkeypox typically presents with small well-localized eschars surrounded by slight erythema, this patient demonstrated well-extended erythema and diffuse eschars on the bilateral upper extremities. This case illustrates that HIV and Monkeypox co-infection may lead to exacerbation of the typical symptoms of each disease– resulting in atypical dermatologic presentations, unique disease manifestations, and more difficult management. 


Rare presentation of Monkeypox in a Patient with Coexisting HIV

Background: SARS-CoV-2 or (CoV2) utilizes the receptor binding domain (RBD) of its surface SPIKE protein to infect human cells that express ACE2 receptor. Infected persons can be asymptomatic or progress to a potentially fatal pneumonia-associated illness. Antibodies that prevent SPIKE/RBD/ACE2 interaction can result from infection or vaccination. However, their quantity, function, and cross-reactive protection requires serological testing. We obtained 50 deidentified serum samples, and were blinded to CoV2 vaccination (Vax+; Vax-) or CoV2 positive test (CoV2+; CoV2-). Serum IgG antibodies to SPIKE and RBD were quantified and efficacy in preventing CoV2 infection in vitro determined. We hypothesized that high titer RBD IgG would be efficient at preventing viral infection and be effective against a later CoV2 variant.

Methods: ELISA was used to quantify SPIKE and RBD antibodies in sera. Neutralizing capability of antibodies was determined by mixing sera with a firefly luciferase lentivirus psuedotyped with Wuhan CoV2 SPIKE. The mixture was applied to ACE2 expressing cells and luciferase activity used to calculate infection rates. Samples were unblinded to determine correlation between in vitro results and CoV2 exposure or vaccination. Cross-protection was determined by repeating neutralization assays with the Omicron BA.2 variant.

Results Thirteen participants reported no CoV2 vaccine or CoV2 positive test (Vax-/CoV2-) yet 7 had anti-SPIKE and -RBD titers and neutralizing function suggesting prior asymptomatic CoV2 infection. The remaining 37 participants reported Vax-/CoV2+ (n=8), Vax+/CoV2- (n=27), or Vax+/CoV2+ (n=2). Titers to RBD were comparatively lower than titers to SPIKE in most patient samples. Average RBD antibody titers were higher for the Vax+/CoV2- group (1:2600) versus Vax-/CoV2+ cohort (1:900). Highest RBD antibody titers were found in Vax+/CoV2+ patients (1:5400). Finally, sera with RBD titer ≥1:2700 and neutralization capacity had equal neutralizing function on Omicron BA.2, a variant with 28 amino acid changes in SPIKE and 16 in the critical RBD region.

Conclusion High-titer RBD sera correlated with greater viral neutralization activity highlighting the functional importance of this domain in infection. High-titer anti-RBD IgG to the initial Wuhan CoV2 virus also provided potent neutralization activity against the BA.2 variant despite 16 amino acid changes in its RBD region. This information could be used in future vaccination efforts and passive immunization, as well as provide a greater level of confidence to healthcare workers, first responders, and other high-risk populations regardless of the CoV2 strain. Serial immune studies in high risk populations could provide indications for booster vaccines.

Functional Serologic Testing and Cross Reactivity of Serum Antibodies to SARS-CoV-2 SPIKE Protein

Background: Cholangiocarcinoma (CCA) is a malignancy of the bile ducts with five-year survival rates of less than 10%. For those with early-stage disease, resection or liver transplantation can be curative. Unfortunately, existing diagnostic tools fail to enable definitive diagnosis, as CCA is notoriously difficult to distinguish from fibrotic or inflammatory lesions. Therefore, novel tools are needed to enable early and accurate diagnosis of CCA, especially in those with indeterminate lesions. Proteases are enzymes that are frequently dysregulated in cancer, including CCA, playing diverse roles in angiogenesis, invasion, and metastasis. We have previously developed a class of peptide-based diagnostic agents, activatable zymography probes (AZPs), which bind to cell membranes after proteolytic cleavage and can be used for ex vivo and in vivo cancer imaging. In this work, we sought to develop AZPs to distinguish CCA from fibrosis.

Methods: We induced CCA in the Mdr2-/- mouse model of biliary fibrosis via hydrodynamic injection of plasmids encoding for AKT, YAP1, and the Sleeping Beauty (SB) transposase. We synthesized a panel of 26 AZPs, applied them to liver tissue sections from CCA and Mdr2-/- mice, and performed whole-slide fluorescence imaging. Select AZPs were co-stained with antibodies against vimentin and cytokeratin 7 and validated in a mouse model of FBXW7/AKT-mutant CCA. Finally, AZPs were PEGylated to increase solubility, intravenously injected into CCA and Mdr2-/- mice, and livers were imaged with in vivo imaging system (IVIS) to assess tumor uptake in vivo. 

Results: We found that 14 AZPs, most notably AZP6, exhibited significantly increased binding to CCA tumors (padj¬ < 0.05, fold change > 1.5) relative to fibrotic bile ducts. In CCA tumors, AZP6 bound significantly more strongly to cells with higher expression of vimentin (p < 0.01) and lower expression of cytokeratin 7 (p < 0.001), suggesting that AZP6 is cleaved by mesenchymal cells. AZP6 also specifically labeled tumors from an FBXW7/AKT-mutant CCA model in a pattern similar to that seen in the YAP1/AKT model, demonstrating its generalizability. Finally, after in vivo administration, AZP6 demonstrated significantly more fluorescence in explanted livers from mice with CCA than mice with fibrosis (p < 0.01), enabling perfect discrimination of malignant and benign disease (AUC = 1, p = 0.009).

Conclusion: AZPs detect and localize dysregulated protease activity ex vivo and in vivo in mouse models of CCA, without false positives from benign biliary fibrosis. With further refinement, AZPs may aid in the diagnosis of CCA.

Detection of cholangiocarcinoma with protease activity probes

High-quality parent language may support preschoolers’ language development by shaping underlying neurodevelopment. Indeed, more interactive language relates to higher frontal brain activity (Romeo et al., 2018). However, it remains to be explored whether other features of parent language may also be related to brain activity. This study examined whether parental linguistic sophistication is related to children’s spontaneous high-frequency neural activity, as indexed by EEG gamma power. Transcripts of parent-child dyads (N=36, children M=4.1 years old) completing a puzzle task were analyzed for mean length of utterance (measure of grammatical complexity) and number of different words (measure of vocabulary sophistication). We found that higher parental mean length of utterance was related to higher regional EEG gamma power (F(2.61,80.78)=2.92, p=.046), suggesting that more grammatically complex parent language is linked with higher child neural maturity. Future studies should explore the directionality of this finding and further measures of parent language quality.

Sophisticated parent language is linked to high-frequency neural activity in preschoolers

A causal illusion occurs when people perceive a causal relationship between two events that are not contingent on each other. How this illusion varies when people reason diagnostically (i.e., in an effect-to-cause direction) is not well known. Thus, this experiment explored how diagnostic reasoning affects causal illusions. Participants learnt about a illusory cause-effect relationship in which the probability of the cause and the probability of the effect were orthogonally manipulated to be either high or low. Participants learnt either predictively (i.e., cause-to-effect) or diagnostically, and at test had to make two causal judgements that encouraged either predictive (cause-to-effect) or diagnostic (effect-to-cause) reasoning. Diagnostic reasoning at test increased the strength of the cause density bias and decreased the strength of the effect density bias. It also decreased causal ratings, but only after predictive learning. These findings highlight the promise of diagnostic reasoning as a tool to debias causal decision making.

Question Framing Modulates the Cause Density and Effect Density Biases in Causal Illusions

Previous researches on multimedia fake news detection include a series of complex feature extraction and fusion networks to gather useful information from the news. However, how cross-modal consistency relates to the fidelity of news and how features from different modalities affect the decision-making are still open questions. This paper presents a novel scheme of Bootstrapping Multi-view Representations (BMR) for fake news detection. Given a multi-modal news, we extract representations respectively from the views of the text, the image pattern and the image semantics. Improved Multi-gate Mixture-of-Expert networks (iMMoE) are proposed for feature refinement and fusion. Representations from each view are separately used to coarsely predict the fidelity of the whole news, and the multimodal representations are able to predict the cross-modal consistency. With the prediction scores, we reweigh each view of the representations and bootstrap them for fake news detection. Extensive experiments conducted on typical fake news detection datasets prove that BMR outperforms state-of-the-art schemes.

Bootstrapping Multi-view Representations for Fake News Detection

In this paper, we develop machine learning techniques to identify unknown printers in early modern (c.~1500--1800) English printed books.
Specifically, we focus on matching uniquely damaged character type-imprints in anonymously printed books to works with known printers in order to provide evidence of their origins.
Until now, this work has been limited to manual investigations by analytical bibliographers.
We present a Contrastive Attention-based Metric Learning approach to identify similar damage across character image pairs, which is sensitive to very subtle differences in glyph shapes, yet robust to various confounding sources of noise associated with digitized historical books. 
To overcome the scarce amount of supervised data, we design a random data synthesis procedure that aims to simulate bends, fractures, and inking variations induced by the early printing process.
Our method successfully improves downstream damaged type-imprint matching among printed works from this period, as validated by in-domain human experts. The results of our approach on two important philosophical works from the Early Modern period demonstrate potential to extend the extant historical research about the origins and content of these books. 

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Concurrent radiation and immunotherapy augments immunity and survival in aneuploid lung cancer

A Potentially Lethal Harbinger: Clostridium Septicum Bacteremia in a Patient Found to have Adenocarcinoma of the Colon

Utilizing Atypical Isolates of Acinetobacter baumannii to Uncover Novel Mechanisms of Resistance

The role of Lcn2 in Triple Negative Breast Cancer Organoids

Effect of mesenchymal stromal cell delivery through cardiopulmonary bypass in a piglet model

Rare presentation of Monkeypox in a Patient with Coexisting HIV

Functional Serologic Testing and Cross Reactivity of Serum Antibodies to SARS-CoV-2 SPIKE Protein

Detection of cholangiocarcinoma with protease activity probes

Sophisticated parent language is linked to high-frequency neural activity in preschoolers

Question Framing Modulates the Cause Density and Effect Density Biases in Causal Illusions

Bootstrapping Multi-view Representations for Fake News Detection

Contrastive Attention Networks for Attribution of Early Modern Print

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