Dynamic Pneumococcal Genetic Adaptations Support Bacterial Growth and Inflammation during Coinfection with Influenza.

Streptococcus pneumoniae (pneumococcus) is one of the primary bacterial pathogens that complicates influenza virus infections. These bacterial coinfections increase influenza-associated morbidity and mortality through a number of immunological and viral-mediated mechanisms, but the specific bacterial genes that contribute to postinfluenza pathogenicity are not known. Here, we used genome-wide transposon mutagenesis (Tn-Seq) to reveal bacterial genes that confer improved fitness in influenza virus-infected hosts. The majority of the 32 genes identified are involved in bacterial metabolism, including nucleotide biosynthesis, amino acid biosynthesis, protein translation, and membrane transport. We generated mutants with single-gene deletions (SGD) of five of the genes identified, SPD1414, SPD2047 (cbiO1), SPD0058 (purD), SPD1098, and SPD0822 (proB), to investigate their effects on in vivo fitness, disease severity, and host immune responses. The growth of the SGD mutants was slightly attenuated in vitro and in vivo, but each still grew to high titers in the lungs of mock- and influenza virus-infected hosts. Despite high bacterial loads, mortality was significantly reduced or delayed with all SGD mutants. Time-dependent reductions in pulmonary neutrophils, inflammatory macrophages, and select proinflammatory cytokines and chemokines were also observed. Immunohistochemical staining further revealed altered neutrophil distribution with reduced degeneration in the lungs of influenza virus-SGD mutant-coinfected animals. These studies demonstrate a critical role for specific bacterial genes and for bacterial metabolism in driving virulence and modulating immune function during influenza-associated bacterial pneumonia.

Enhanced Cancer Immunotherapy by Microneedle Patch-Assisted Delivery of Anti-PD1 Antibody.

Despite recent advances in melanoma treatment through the use of anti-PD-1 (aPD1) immunotherapy, the efficacy of this method remains to be improved. Here we report an innovative self-degradable microneedle (MN) patch for the sustained delivery of aPD1 in a physiologically controllable manner. The microneedle is composed of biocompatible hyaluronic acid integrated with pH-sensitive dextran nanoparticles (NPs) that encapsulate aPD1 and glucose oxidase (GOx), which converts blood glucose to gluconic acid. The generation of acidic environment promotes the self-dissociation of NPs and subsequently results in the substantial release of aPD1. We find that a single administration of the MN patch induces robust immune responses in a B16F10 mouse melanoma model compared to MN without degradation trigger or intratumoral injection of free aPD1 with the same dose. Moreover, this administration strategy can integrate with other immunomodulators (such as anti-CTLA-4) to achieve combination therapy for enhancing antitumor efficacy.

Synchronous tracheostomy and gastrostomy placement results in shorter length of stay in traumatic brain injury patients.

BACKGROUND: American Society for Parenteral and Enteral Nutrition (ASPEN) guidelines recommend gastrostomy for patients suspected to require enteral access device for 4-6 weeks. Our hypothesis was that traumatic brain injury (TBI) patients undergoing synchronous tracheostomy/gastrostomy (SYNC) compared to tracheostomy first (DELAY) have shorter length of stay (LOS) but higher rates of unnecessary gastrostomy. METHODS: Retrospective review of TBI patients requiring tracheostomy in 2017-2022 â€‹at a Level 1 trauma center was conducted. SYNC and DELAY patients were compared, and CoxPH analysis was performed for LOS. RESULTS: 394 patients were included [mean age: 42 (SD:18); mortality: 9 â€‹%]. The DELAY group had longer LOS (39 vs 32 days, p â€‹< â€‹0.001). There was no significant difference in unnecessary gastrostomy rate between groups (p â€‹= â€‹0.1331). In adjusted hazard analysis, SYNC predicted shorter LOS (HR:1.54; 95 â€‹% CI:1.20-1.98, p â€‹< â€‹0.001). CONCLUSIONS: Synchronous gastrostomy was associated with shorter length of stay and similar rates of unnecessary gastrostomy in TBI patients.

Misinterpretations of Significance Testing Results Near the P-Value Threshold in the Urologic Literature.

Background The outcome of a statistical test is to accept or reject a null hypothesis. Reporting a metric as "trending toward significance" is a misinterpretation of the p-value. Studies highlighting the prevalence of statistical errors in the urologic literature remain scarce. We evaluated abstracts from 15 urology journals published within the years 2000-2021 and provided a quantitative measure of a common statistical mistake-misconstruing the function of null hypothesis testing by reporting "a trend toward significance." Materials and methods We performed an audit of 15 urology journals, looking at articles published from January 1, 2000, to January 1, 2022. A word recognition function in Microsoft Excel was utilized to identify the use of the word "trend" in the abstracts. Each use of the word "trend" was manually investigated by two authors to determine whether it was improperly used in describing non-statistically significant data as trending toward significance. Statistics and data analysis were performed using Python libraries: pandas, scipy.stats, and seaborn. Results This study included 101,134 abstracts from 15 urology journals. Within those abstracts, the word "trend" was used 2,509 times, 572 uses of which were describing non-statistically significant data as trending toward significance. There was a statistically significant difference in the rate of errors between the 15 journals (p < 0.01). The highest rate of improper use of the word "trend" was found in Bladder Cancer with a rate of 1.6% (p < 0.01) of articles. The lowest rate of improper use was found in European Urology, with a rate of 0.3% (p < 0.01). Our analysis found a moderate correlation between the number of articles published and the number of misuses of the word "trend" within each journal and across all journals every year (r = 0.61 and 0.70, respectively). Conclusion The overall rate of p-value misinterpretation never exceeded 2% of articles in each journal. There is significance in the difference in misinterpretation rates between the different journals. Authors' utilization of the word "trend" describing non-significant p-values as being near significant should be used with caution.

Transformable DNA nanocarriers for plasma membrane targeted delivery of cytokine.

Direct delivery of cytokines using nanocarriers holds great promise for cancer therapy. However, the nanometric scale of the vehicles made them susceptible to size-dependent endocytosis, reducing the plasma membrane-associated apoptosis signaling. Herein, we report a tumor microenvironment-responsive and transformable nanocarrier for cell membrane targeted delivery of cytokine. This formulation is comprised of a phospholipase A2 (PLA2) degradable liposome as a shell, and complementary DNA nanostructures (designated as nanoclews) decorated with cytokines as the cores. Utilizing the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) as a model cytokine, we demonstrate that the TRAIL loaded DNA nanoclews are capable of transforming into nanofibers after PLA2 activation. The nanofibers with micro-scaled lengths efficiently present the loaded TRAIL to death receptors on the cancer cell membrane and amplified the apoptotic signaling with reduced TRAIL internalization.

Synergistic Transcutaneous Immunotherapy Enhances Antitumor Immune Responses through Delivery of Checkpoint Inhibitors.

Despite the promising efficacy of immunoregulation in cancer therapy, the clinical benefit has been restricted by inefficient infiltration of lymphocytes in the evolution of immune evasion. Also, immune-related adverse events have often occurred due to the off-target binding of therapeutics to normal tissues after systematic treatment. In light of this, we have developed a synergistic immunotherapy strategy that locally targets the immunoinhibitory receptor programmed cell death protein 1 (PD1) and immunosuppressive enzyme indoleamine 2,3-dioxygenase (IDO) for the treatment of melanoma through a microneedle-based transcutaneous delivery approach. The embedded immunotherapeutic nanocapsule loaded with anti-PD1 antibody (aPD1) is assembled from hyaluronic acid modified with 1-methyl-dl-tryptophan (1-MT), an inhibitor of IDO. This formulation method based on the combination strategy of "drug A in carriers formed by incorporation of drug B" facilitates the loading capacity of therapeutics. Moreover, the resulting delivery device elicits the sustained release and enhances retention of checkpoint inhibitors in the tumor microenvironment. Using a B16F10 mouse melanoma model, we demonstrate that this synergistic treatment has achieved potent antitumor efficacy, which is accompanied by enhanced effective T cell immunity as well as reduced immunosuppression in the local site.