Pic-Producing Escherichia coli Induces High Production of Proinflammatory Mediators by the Host Leading to Death by Sepsis.

Escherichia coli is an important pathogen responsible for a variety of diseases. We have recently shown that Pic, a serine protease secreted by E. coli, mediates immune evasion by the direct cleavage of complement molecules. The aim of this study was to investigate the action of a Pic-producing bacteria in a murine model of sepsis. Mice were infected with Pic-producing E. coli (F5) or F5∆pic mutant. Animal survival was monitored for five days, and a subset of mice was euthanized after 12 h for sample acquisition. The inoculation of Pic-producing bacteria induced 100% death within 24 h. The colony forming units count in the organs was significantly higher in F5. Hematological analysis showed a decrease of total leukocytes. Nitric oxide and cytokines were detected in serum, as well as on peritoneal lavage of the F5 group in higher levels than those detected in the other groups. In addition, immunophenotyping showed a decrease of activated lymphocytes and macrophages in the F5 group. Therefore, Pic represents an important virulence factor, allowing the survival of the bacterium in the bloodstream and several organs, as well as inducing a high production of proinflammatory mediators by the host, and concomitantly a cellular immunosuppression, leading to sepsis and death.

Prophylactic use of standardized extract of propolis of Apis mellifera (EPP-AF®) reduces lung inflammation and improves survival in experimental lethal sepsis.

ETHNOPHARMACOLOGICAL RELEVANCE: Sepsis poses one of the biggest public health problems, necessitating the search for new therapeutic alternatives. For centuries, propolis has been widely used in folk medicine to treat various inflammatory and infectious diseases. Given its extensive use, it has excellent potential as an adjuvant treatment for patients with sepsis. OBJECTIVE: This study evaluated prophylactic treatment with standardized propolis extract (EPP-AF®) and followed the prognosis of sepsis induced by ligation and cecal ligation and puncture (CLP). METHODS: Initially, for survival assessment, Swiss mice were separated into five groups: Sham (false operated), control (PBS), ATB (received antibiotic, 8 mg/kg), P10 (received EPP-AF®, 10 mg/kg), and P100 (received EPP-AF®, 100 mg/kg). The animals received PBS, antibiotic, or EPP-AF® by the subcutaneous route 6 h before the CLP procedure. Animal survival was assessed every 12 h for five days when all of them were euthanized. RESULTS: We show that the treatment with EPP-AF® significantly increased the life expectancy of animals with sepsis compared to the control group. Interestingly, prophylactic treatment with EPP-AF® showed no effect on the number of colony-forming units in the peritoneum, blood, or lung. However, there was a decrease in cellular influx in the peritoneum. This alteration was unrelated to the number of bone marrow cells or the differential counting of peripheral blood cells. The coagulogram remained unchanged, including the number of platelets and prothrombin time-activated partial thromboplastin time. However, the inflammatory infiltrate and bleeding in the lung tissue were lower in the animals that received EPP-AF®. CONCLUSION: Thus, it was possible to conclude that prophylactic treatment with EPP-AF® preserved the lung parenchyma, resulting in an increased lifespan of mice with sepsis. It can be a helpful adjuvant in prophylactic treatment with antibiotics in presurgical conditions.

MIR-376A-2-5P as a potential prognostic marker for advanced penile squamous cell carcinomas trough HPV-dependent pathways.

Penile cancer (PeCa) is a rare tumor, generally associated with socioeconomic conditions in low-income countries. Hence, a delay in diagnosis and treatment leads in more advanced tumors, to higher comorbidity, and mortality. Human papillomavirus (HPV) infection has been identified as one of the major risk factors for PeCa. In addition, viral integration sites have been related to copy number alterations, impacting miRNAs/mRNA interactions and, consequently, the molecular pathways related to them. Nonetheless, studies on differentially expressed miRNAs (miRDEs) in PeCa are still scarce, especially in PeCa associated with high-risk HPV (hrHPV). To investigate the role of these gene regulators in PeCa progression, 827 miRNAs (Nanostring Technologies™, Seattle, WA, USA) were evaluated in 22 hrHPV-associated penile squamous cell carcinomas and five non-tumor penile tissues. For functions of miRNAs/target genes and relationship with HPV we conducted an integrated analysis by Diana Tools, KEGG, HPVbase, and InterSPPI-HVPPI platforms. We found that 25 miRNAs of the most differentially expressed impact 43 top molecular pathways, of which the fatty acid biosynthesis pathway, prions, miRNAs in cancer and hippo signaling (P<1.0-325, for each) were the most statistically significant. Notably, 23 out of 25 are located at HPV integration sites (HPVis). MiR-1206, miR-376b-3p and miR-495-3p were downregulated and associated with perineural invasion. In addition, a comparison between advanced and early diseases revealed 143 miRDEs. ROC analysis of a single (miR-376a-2-5p), paired (miR-376a-2-5p, miR-551b-3p) or combination of five miRDEs (miR-99a-5p, miR-150-5p, miR-155-5p, let-7c-5p, miR-342-3p) showed robust discriminatory power (AUC = 0.9; P = 0.0114, for each). Strikingly, miR-376a-2-5p exhibited the highest values of sensitivity and specificity, with 100% and 83.3%, respectively, indicating this miRNA as a potential prognostic marker in hrHPV-penile carcinogenesis.

The Immunomodulatory Activity of Punica granatum L. Peel Extract Increases the Lifespan of Mice with Lethal Sepsis.

Sepsis is an organ dysfunction syndrome associated with high mortality. To date, no effective treatment is available to combat this disease. Punica granatum L. is a potential alternative treatment due to its anti-inflammatory, antimicrobial, and antioxidant properties. Thus, this study aimed to evaluate the effects of a hydroalcoholic crude extract from the peels of P. granatum (HCEPg) in mice with lethal sepsis. Lethal polymicrobial sepsis was induced in female Swiss mice via cecal ligation and puncture (CLP). Initially, the animals were divided into three groups: Sham (false-operated), CLP-control (phosphate-buffered saline), and CLP-HCEPg (single dose, 5 mg/kg, subcutaneous administration). Treatment was initiated immediately after the induction of sepsis, and survival was evaluated every 12 hr for 5 days. Those who survived were euthanized. Serum cytokine levels were measured using a cytometric bead array Mouse Inflammatory Cytokine Kit. The number of colony-forming units, as well as the number of cells in the lymphoid organs and their activation markers, were analyzed. Results showed that treatment with HCEPg increased lifespan and reduced bacterial counts in the peritoneum, bloodstream, and spleen. HCEPg also decreased hydrogen peroxide secretion by phagocytes and augmented serum IL-10 levels, indicating its systemic anti-inflammatory effects. Additionally, treatment with HCEPg attenuated infection-induced lung hemorrhage. Overall, P. granatum extract improved the lifespan of septic mice, possibly due to its antimicrobial, anti-inflammatory, and immunomodulatory effects, thereby regulating bacterial load and translocation, as well as controlling the systemic inflammation induced by sepsis.

Neutrophil-to-Lymphocyte and Platelet-to-Lymphocyte Ratios in Nondialysis Chronic Kidney Patients.

BACKGROUND: The Neutrophil-to-Lymphocyte Ratio (NLR) and the Platelet-to-Lymphocyte Ratio (PLR) are inflammatory biomarkers for several diseases, such as cancer and cardiovascular morbidities; however, there are currently few studies on kidney diseases. We aimed to evaluate nondialysis patients and determine the association of NLR and PLR with inflammation in these patients. METHODS: A prospective cross-sectional study was conducted with 85 patients at different stages of chronic kidney disease (CKD), treated at the Kidney Disease Prevention Center of the University Hospital of the Federal University of Maranhão. This study included adult nondialysis patients diagnosed with CKD. The participants' blood samples were collected for a high-sensitivity C-reactive protein (hs-CRP) test and blood count. They were divided into two groups according to the presence or absence of inflammation based on the hs-CRP value (<0.5 mg/dL). NLR and PLR were calculated based on the absolute number of neutrophils, lymphocytes, and platelets and were compared between them and with hs-CRP. Statistical analysis was performed using the Stata software, with the Shapiro-Wilk, Mann-Whitney, Spearman's Correlation, and receiver operating characteristic curve tests. This study was approved by the local ethics committee. RESULTS: The participants were categorized into two groups: with inflammation (n = 64) and without inflammation (n = 21). The mean age was 61.43 ± 14.63 y. The NLR and PLR values were significantly different between the groups with and without inflammation (p=0.045and p=0.004, respectively). However, only PLR showed a significant positive correlation with hs-CRP (p=0.015). The best cutoff point for NLR to detect inflammation was 1.98, with 76.19% sensitivity and 48.44% specificity. For PLR, it was 116.07, with 85.71% sensitivity and 51.56% specificity. There was no significant difference between the area under the NLR and PLR curve (0.71 vs. 0.64; p=0.186) for this population. CONCLUSIONS: This study showed that PLR was positively correlated with hs-CRP in nondialysis CKD patients and can be used to identify inflammation in this population.

Preparation, characterization and in vitro anticancer performance of nanoconjugate based on carbon quantum dots and 5-Fluorouracil.

This study is focused on the development of a nanodevice for loading and release of 5-Fluorouracil (5-FU) with a view to improving its therapeutic efficiency, using as strategy the fabrication of a nanoconjugate through drug anchorage on the surface of carbon quantum dots (CQD). Several physicochemical and analytical techniques were employed to obtain information about materials morphology, structure, and optical properties. The results indicated that the interactions between both entities resulted in good physicochemical properties and photostability. Acid pH favored drug release, indicating a tendency to release 5-FU from 5-FU-CQD into the tumor microenvironment. The cytotoxicity of CQD and 5-FU-CQD nanoconjugate was evaluated against normal human lung fibroblast (GM07492A) and human breast cancer (MCF-7) cell lines. The CQD was non-toxic, indicating that these materials are biocompatible and can be used as a nanocarrier for 5-FU in biological systems. For the 5-FU-CQD nanoconjugate, it was observed a reduction in toxicity for normal cells compared to free 5-FU, suggesting that drug anchoring in CQD reduced drug-associated toxicity, while for cancer cells exhibited an antitumor effect equivalent to that of the free drug, opening perspectives for the application of this material in anticancer therapy.