Viral defense at mucosal sites depends on interferons (IFN) and IFN stimulated genes (ISGs), either of which may be constitutively expressed to maintain an “antiviral state” (AVS). However, the mechanisms that govern the AVS are poorly defined. Using a BEAS-2B respiratory epithelial cell line deficient in IRF1, we demonstrate higher susceptibility to infection with vesicular stomatitis virus (VSV) and influenza virus. IRF1-mediated restriction of VSV is IFN-independent, as blockade of types I and III IFNs and JAK-STAT signaling before infection did not affect VSV infection of either parent or IRF1 KO cells. Transcriptome analysis revealed that IRF1 regulates constitutive expression of ~300 genes, including antiviral ISGs: OAS2, BST2 , and RNASEL and knockdown of any of these IRF1-dependent genes increased VSV infection. Additionally, IRF1 enhances rapid expression of IFNβ and IFNλ after stimulation with poly I:C and also regulates ISG expression. Mechanistically, IRF1 enhances recruitment of BRD4 to promotor-enhancer regions of ISGs for rapid expression and maintains levels of histone H3K4me1 for optimal constitutive expression. Finally, IRF1 also regulates constitutive expression of TLR2 and TLR3 and promotes signaling through these pattern recognition receptors (PRR). These data reveal multiple roles for IRF1 toward effective anti-viral responses by maintaining IFN-independent constitutive expression of anti-viral ISGs and supporting early IFN-dependent responses to PRR stimulation.

After viral infection, type I and III interferons (IFNs) are coexpressed by respiratory epithelial cells (RECs) and activate the ISGF3 transcription factor (TF) complex to induce expression of a cell-specific set of interferon-stimulated genes (ISGs). Type I and III IFNs share a canonical signaling pathway, suggesting that they are redundant. Animal and in vitro models, however, have shown that they are not redundant. Because TFs dictate cellular phenotype and function, we hypothesized that focusing on TF-ISG will reveal critical combinatorial and nonredundant functions of type I or III IFN. We treated BEAS-2B human RECs with increasing doses of IFNβ or IFNλ1 and measured expression of TF-ISG. ISGs were expressed in a dose-dependent manner with a nonlinear jump at intermediate doses. At subsaturating combinations of IFNβ and IFNλ1, many ISGs were expressed in a pattern that we modeled with a cubic equation that mathematically defines this threshold effect. Uniquely, IFNβ alone induced early and transient IRF1 transcript and protein expression, while IFNλ1 alone induced IRF1 protein expression at low levels that were sustained through 24 h. In combination, saturating doses of these 2 IFNs together enhanced and sustained IRF1 expression. We conclude that the cubic model quantitates combinatorial effects of IFNβ and IFNλ1 and that IRF1 may mediate nonredundancy of type I or III IFN in RECs.

Examined effect of long wait-time (≥8 weeks) for surgical treatment on survival of women with early-stage cervical cancer.• Long surgery wait-time was not associated with short-term disease recurrence of women with early-stage cervical cancer.• Systematic review suggests that surgery wait-time for up to 8 weeks may have limited/modest effect on disease recurrence.

Micro(mi)RNAs are 21-to 23-nt RNAs that regulate multiple biological processes. In association with Argonaute (Ago) proteins and other factors that form the RNA-induced silencing complex (RISC), miRNAs typically bind mRNA 3 ′ untranslated regions (UTRs) and repress protein production through antagonizing translation and transcript stability. For a given mRNA-miRNA interaction, cis-acting RNA elements and trans-acting RNA-binding proteins (RBPs) may influence mRNA fate. This is particularly true of the hepatitis C virus (HCV) genome which interacts with miR-122, an abundant liver miRNA. miR-122 binding to HCV RNA considerably stimulates virus replication in cultured cells and primates, but the mechanism(s) and associated host factors required for enhancement of HCV replication have not been fully elucidated. We recapitulated miR-122-HCV RNA interactions in a cell-free translation system derived from cells that express miR-122. Specifically, lysates produced from HEK-293 cells that inducibly transcribe and process pri-miR-122 were characterized alongside those from isogenic cells lacking miR-122 expression. We observed a stimulatory effect of miR-122 on HCV reporter mRNAs in a manner that depended on expression of miR-122 and intact target sites within the HCV 5 ′ UTR. We took advantage of this system to affinity-purify miR-122-HCV RNP complexes. Similar to functional assays, we found that association of immobilized HCV internal ribosome entry site (IRES) RNA with endogenous Ago2 requires both miR-122 expression and intact miR-122 target sites in cis. This combined approach may be generalizable to affinity purification of miRNP complexes for selected target mRNAs, allowing identification of miRNP components and RBPs that may contribute to regulation.

Targeted reduction in IgE reactivity of native allergen extracts to produce allergoids via covalent cross-linking is beneficial in producing safe and efficacious immunotherapies. We present techniques to demonstrate the presence of the relevant allergens in allergoid preparations by tandem mass spectrometry and the molecular fingerprint of those allergoids by high performance-size exclusion chromatography (HPLC-SEC). METHODS: The polymerization profile of sweet grass allergoids was determined by HPLC-SEC, from which separate size fractions of allergoids were collected. Proteomic analysis of each corresponding fraction was purified, subject to tryptic digest and analysed via tandem mass spectrometry. Once the peptide had been identified, it was compared to protein databases such as NCBI or SwissProt, from which the sequence identity was assessed. RESULTS: HPLC-SEC highlighted the spread of allergens/allergoids preand post-modification. The HPLC profiles of the allergoids showed a decrease in retention time (increase in molecular weight) after modification (i.e. polymerization). A greater number of allergens are identified from tandem mass spectrometry (proteomic) analysis as the predicted molecular weight range of each fraction decreases. CONCLUSIONS: Native and modified extracts are not two discrete preparations but are instead a formula of native and modified allergens, within which IgG reactive epitopes are present. Proteomic analysis confirmed the presence of allergens from multiple grass species. This work demonstrates that IgG epitopes remain in an allergoid formulation while IgE epitopes are attenuated, allowing safe administration of a higher strength product in fewer doses.

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INTRODUCTION: The incidence of STIs, specifically gonorrhea, chlamydia, and trichomoniasis among Los Angeles County residents is on the rise. One possible explanation is inadequate treatment, especially of sexual partners. Expedited partner therapy (EPT) is a method of treating patients' partners without a separate healthcare visit, which has been shown to decrease persistent and/or recurrent infections. We report on quality improvement outcomes following interventions to help increase the rates of EPT provision. METHODS: Interventions included creating a protocol with the pharmacy, arranging for Cefixime to be on formulary, and conducting resident training sessions. Data collection was performed on all positive gonorrhea (GC), chlamydia (CT) and trichomoniasis lab results seen from January 1, 2017 to February 27, 2019 across all departments, inclusive of Ob/Gyn. EPT rates before and after interventions are presented. RESULTS: The rates of EPT prescriptions provided by OB/GYN providers for STI treatment increased following the interventions. EPT for GC/CT increased from 40% to 55% of cases; EPT for Trichomonas increased from 38% to 85%. Interestingly, patients were often counseled on the need for partner treatment, but were not given prescriptions. This study includes only female patients seen by Ob/Gyn providers, though the majority of results were detected in the emergency department by non Ob/Gyn providers. CONCLUSION: Following interventions aimed at establishing agreements with the pharmacy as well as increasing awareness among residents, the EPT rates of gonorrhea, chlamydia, and trichomoniasis did increase significantly. This may assist with decreasing the rate of STI reinfection among County hospitals and hospitals supported by medical trainees.

Purpose: To identify patient factors associated with medical management failure and hospital readmission among patients admitted with a diagnosis of tuboovarian abscess at a public hospital in Los Angeles, California. Methods: We conducted a retrospective chart review of admissions for tuboovarian abscess from January 2014 through August 2019, describing treatment plans and their subsequent outcomes. Patient factors associated with failure of medical management, as defined by requiring image-guided drainage or surgery, and hospital readmission were identified via bivariate analyses, using Chi-square, Fisher’s exact, and Wilcoxon signed-rank tests as appropriate. Factors independently linked to failure of medical management were determined via logistic regression. Results: We treated 147 patients with tuboovarian abscess over 5 years, comprised primarily of Hispanic/Latinx (74%; n=109/147) women, median age 39 (range: 18-70) years. The median abscess size was 6.3cm (range: 4.8-8.0). Only 16% (n=23/147) who initially received parenteral antibiotic therapy failed medical management. Factors independently and positively linked to parenteral antibiotic failure included elevated white blood cell count (aOR 1.17; 95%CI 1.04-1.32) and abscess size (aOR 1.07; 95%CI 1.04-1.10). No tuboovarian abscesses under 5.5 centimeters required procedural intervention when treated with parenteral antibiotics alone. We identified rehospitalizations among 13.6% (n=20/147) of patients; no factors were significantly linked to rehospitalization.Conclusions: Tuboovarian abscesses were successfully treated with medical management in more than 80% of cases. Early procedural intervention for tuboovarian abscesses greater than 5.5 centimeters in diameter may help to avoid short and long-term morbidities.