BacScan: a novel genome-wide strategy for uncovering broadly immunogenic proteins in bacteria.

In response to the global threat posed by bacterial pathogens, which are the second leading cause of death worldwide, vaccine development is challenged by the diversity of bacterial serotypes and the lack of immunoprotection across serotypes. To address this, we introduce BacScan, a novel genome-wide technology for the rapid discovery of conserved highly immunogenic proteins (HIPs) across serotypes. Using bacterial-specific serum, BacScan combines phage display, immunoprecipitation, and next-generation sequencing to comprehensively identify all the HIPs in a single assay, thereby paving the way for the development of universally protective vaccines. Our validation of this technique with Streptococcus suis, a major pathogenic threat, led to the identification of 19 HIPs, eight of which conferred 20-100% protection against S. suis challenge in animal models. Remarkably, HIP 8455 induced complete immunity, making it an exemplary vaccine target. BacScan's adaptability to any bacterial pathogen positions it as a revolutionary tool that can expedite the development of vaccines with broad efficacy, thus playing a critical role in curbing bacterial transmission and slowing the march of antimicrobial resistance.

De novo identification of bacterial antigens of a clinical isolate by combining use of proteosurfaceomics, secretomics, and BacScan technologies.

Background: Emerging infectious diseases pose a significant threat to both human and animal populations. Rapid de novo identification of protective antigens from a clinical isolate and development of an antigen-matched vaccine is a golden strategy to prevent the spread of emerging novel pathogens. Methods: Here, we focused on Actinobacillus pleuropneumoniae, which poses a serious threat to the pig industry, and developed a general workflow by integrating proteosurfaceomics, secretomics, and BacScan technologies for the rapid de novo identification of bacterial protective proteins from a clinical isolate. Results: As a proof of concept, we identified 3 novel protective proteins of A. pleuropneumoniae. Using the protective protein HBS1_14 and toxin proteins, we have developed a promising multivalent subunit vaccine against A. pleuropneumoniae. Discussion: We believe that our strategy can be applied to any bacterial pathogen and has the potential to significantly accelerate the development of antigen-matched vaccines to prevent the spread of an emerging novel bacterial pathogen.

Melatonin blunts the tumor-promoting effect of cancer-associated fibroblasts by reducing IL-8 expression and reversing epithelial-mesenchymal transition.

BACKGROUND: Most studies on melatonin have focused on tumor cells but have ignored the tumor microenvironment (TME), especially one of its important components, the cancer-associated fibroblasts (CAFs). Therefore, we attempted to explore the role of melatonin in TME. METHODS: We investigated the regulatory role of melatonin in the tumor-promoting effect of CAFs and its underlying mechanism by using cell and animal models. RESULTS: CAFs promoted tumor progression, but melatonin weakened the tumor-promoting effect of CAFs. Compared with tumor cells, IL-8 was mainly expressed in CAFs. CAFs-overexpressing IL-8 induced the epithelial-mesenchymal transition (EMT) of tumor cells, and a positive crosstalk was observed between CAFs and tumor cells undergoing EMT, thereby further promoting the IL-8 expression. Melatonin suppressed this crosstalk by inhibiting the NF-κB pathway, thereby impeding the IL-8 expression from CAFs. Importantly, melatonin reversed CAFs-derived IL-8-mediated EMT by inhibiting the AKT pathway. Melatonin was found to directly and indirectly inhibit tumor progression. CONCLUSION: Our research reveals the potential action mechanism of melatonin in regulating the CAF-tumor cell interaction and suggests the potential of melatonin as an adjuvant of tumor therapy.

Expression of the prognostic marker IL-8 correlates with the immune signature and epithelial-mesenchymal transition in breast cancer.

BACKGROUND: IL-8 has been implicated in the malignant progression of various types of cancers; however, the precise molecular mechanisms associated with IL-8 in breast cancer (BRCA) are unclear. METHODS: We analyzed the clinical signature and immune characteristics of BRCA patients and its correlation with IL-8 expression using The Cancer Genome Atlas (TCGA) datasets. The role of IL-8 in epithelial-mesenchymal transition (EMT) was verified through Western blotting, Cell Counting Kit-8 assay, and wound healing assays, as well as cell invasion experiments. RESULTS: Through a comprehensive bioinformatics study, we determined that high IL-8 expression was associated with poor prognosis. Enrichment analysis revealed that high IL-8 expression was enriched in immune-related processes and cancer-related signaling pathways. In addition, IL-8 was associated with most of the immune-infiltrating cells, and high IL-8 expression indicated poor response to immunotherapy. Importantly, we found that IL-8 induced EMT in vitro. CONCLUSIONS: Taken together, our data indicate that IL-8 may be a potential and valuable prognostic marker in BRCA, which may induce adverse outcomes by modulating the immune response and promoting EMT in BRCA patients.

Peritumoral tertiary lymphoid structure and tumor stroma percentage predict the prognosis of patients with non-metastatic colorectal cancer.

Background: Tertiary lymphoid structures (TLSs) are crucial in promoting and maintaining positive anti-tumor immune responses. The tumor stroma has a powerful immunosuppressive function that could exclude tumor-infiltrating lymphocytes from the tumor beds and lead to a "cold" phenotype. TLSs and tumor stroma percentage (TSP) are significantly associated with the prognosis of patients with certain cancers. However, the exact roles of TLSs and TSP and their intrinsic relationship are still largely unknown in colorectal cancer (CRC). Methods: TLSs and TSP were assessed using hematoxylin-eosin (H&E) and/or immunohistochemistry (IHC) staining from 114 CRC patients in the training set and 60 CRC patients in the external validation set. The correlation between TILs, TLS and clinicopathological characteristics and their prognostic values were assessed. Finally, we plotted a Nomogram including the TLS, TSP and tumor-node-metastasis (TNM) stage to predict the probability of recurrence-free survival (RFS) at 2- and 5-years in non-metastatic colorectal cancer (nmCRC) patients. Results: Peritumoral TLS (P-TLS), intratumoral TLS (In-TLS) and high TSP (H-TSP, >50%) were present in 99.1%, 26.3% and 41.2% patients, respectively. H-TSP tumor tends to be associated with lower P-TLS density (P =0.0205). The low P-TLS density (< 0.098/mm2) was significantly associated with reduced RFS (HR=6.597 95% CI: 2.882-15.103, P <0.001) and reduced overall survival (OS) (HR=6.628 95% CI: 2.893-15.183, P < 0.001) of nmCRC patients. In-TLS was not of significance in evaluating the clinical outcomes of nmCRC patients. H-TSP was significantly associated with reduced RFS (HR=0.126 95% CI: 0.048-0.333, P <0.001) and reduced OS (HR=0.125 95% CI: 0.047-0.332, P <0.001) of nmCRC patients. The 5-year RFS of the high P-TLS, low-TLS, H-TSP, and L-TSP groups were 89.7%, 47.2%, 53.2%, and 92.5%, respectively. The P-TLS density, TSP and TNM stage were independent prognosis factors of nmCRC patients. The Nomogram, including the P-TLS density, TSP and TNM stage, outperformed the TNM stage. Conclusions: High P-TLS density and low TSP (L-TSP) were independent and favorable prognostic factors of nmCRC patients, which might provide new directions for targeted therapy in the CRC tumor microenvironment, especially the tumor immune microenvironment.

Short-term efficacy and safety of a lower dose of polyethylene glycol recombinant human growth hormone in children with growth hormone deficiency: A randomized, dose-comparison study.

Objective: Polyethylene glycol recombinant human growth hormone (PEG-rhGH, Jintrolong®) is the first long-acting rhGH preparation that is approved to treat children with growth hormone deficiency (GHD) in China. Clinical experience with dose selections of PEG-rhGH is scarce. The present study compared the efficacy and safety of a lower dose to increase dosing regimens of PEG-rhGH treatment. Methods: A multicenter, randomized, open-label, dose-comparison clinical study was conducted to compare the improvements in the height standard deviation score (Ht SDS), height velocity (HV), insulin-like growth factor-1 (IGF-1) SDS, and safety profiles of children with GHD who are treated with 0.2 mg/kg/week of PEG-rhGH dose or 0.14 mg/kg/week for 26 weeks. Results: Ht SDS, HV, and IGF-1 SDS increased significantly after PEG-rhGH treatment in the two dose groups (p < 0.05). The improvements of Ht SDS, HV, and IGF-1 SDS were more significant in the high-dose group than in the low-dose group (p < 0.05). Ht SDS improvement in low-dose group was not non-inferiority to that in the high-dose group (p = 0.2987). The incidences of adverse events were comparable between the two groups. Conclusion: The improvements of Ht SDS, HV, and IGF-1 SDS were more significant in the high-dose group than in the low-dose group (p < 0.05). PEG-rhGH at the dose of 0.14 mg/kg/week was effective and safe for children with GHD. Clinical Trial Registration: clinicaltrials.gov, identifier NCT02908958.

Pressure ulcers in patients with diabetes: a bibliometrics analysis.

BACKGROUND: Pressure ulcers (PU) refer to local tissue ulceration and necrosis caused by long-term compression and friction brought on by tissue ischemia and hypoxia. Diabetic wounds do not easily heal, and once a pressure ulceration occurs, it is difficult to deal with. The purpose of this study was to analyze the current research status of PUs in diabetic patients. METHODS: The Science Citation Index Expanded (SCI-E) database was searched with terms of "Pressure Ulcer" and "Diabetes". Citespace software was used to analyze the annual distribution of the number of target documents and the distribution of countries, institutions, journals, authors, and keywords used in these works. RESULTS: In all, 1271 documents were retrieved, with a total citation frequency of 47,081, and an h-index of 101. The top 5 countries in terms of the number of publications were the United States, the United Kingdom, China, Australia, and the Netherlands; the top 5 countries in centrality were the Netherlands, the United States, Canada, Japan, and France. The institutions with the greatest number of publications were the University of Amsterdam, Cardiff University, The University of Washington, and the University of Manchester. The institutions with the highest centrality were the University of Amsterdam, the University of Groningen, the University of Washington, the University of Adelaide, Baylor College of Medicine, and Queensland University of Technology. The authors with a highest number of publications were Bus SA, Apelqvist J, and the International Working Group on the Diabetic Foot, and Hinchliffe RJ. Only 2 authors had a centrality score above 0.01. Journals such as Diabetes Metabolism Research and Reviews, Diabetes Care, and Journal of Wound Care showed considerable influence in this field. Keyword analysis indicated that the use of keywords in this field is not uniform, and the focus of research on PUs in diabetic patients lies the risk and management of foot ulcers. CONCLUSIONS: There are few studies concerning PUs in patients with diabetes and little collaboration between authors. The current focus in this field is on the risk and management of foot ulcers.

Two-photon scattering and correlation in a four-terminal waveguide system.

Scattering and correlation properties of a two-photon (TP) pulse are studied in a four-terminal waveguide system, i.e., two one-dimensional waveguides connected by a Jaynes-Cummings emitter (JCE). The wave function approach is utilized to exactly calculate the real-time dynamic evolution of the TP transport. When the width of the incident TP Gaussian pulse is much larger than the photon wavelength, the TP transmission spectra approach that of the corresponding single photon cases and are almost independent of the pulse width. On the contrary, as the pulse width is comparable to the photon wavelength, the TP transmission and correlation both show strong dependence on the pulse width. The resonant scattering due to the JCE and the photon interference together determine the TP correlation. When the distance between the TPs is small, the TP correlations between any two terminals for the scattered TP pulse are much different from those for the incident TP pulse and therefore, such a four-terminal waveguide system provides a way to control the TP correlation.

Engineering T4 Bacteriophage for In Vivo Display by Type V CRISPR-Cas Genome Editing.

Bacteriophage T4 has enormous potential for biomedical applications due to its large size, capsid architecture, and high payload capability for protein and DNA delivery. However, it is not very easy to genetically engineer its genome heavily modified by cytosine hydroxymethylation and glucosylation. The glucosyl hydroxymethyl cytosine (ghmC) genome of phage is completely resistant to most restriction endonucleases and exhibits various degrees of resistance to CRISPR-Cas systems. Here, we found that the type V CRISPR-Cas12a system, which shows efficient cleavage of ghmC-modified genome when compared to the type II CRISPR-Cas9 system, can be synergistically employed to generate recombinant T4 phages. Focused on surface display, we analyzed the ability of phage T4 outer capsid proteins Hoc (highly antigenic outer capsid protein) and Soc (small outer capsid protein) to tether, in vivo, foreign peptides and proteins to T4 capsid. Our data show that while these could be successfully expressed and displayed during the phage infection, shorter peptides are present at a much higher copy number than full-length proteins. However, the copy number of the latter could be elevated by driving the expression of the transgene using the strong T7 RNA polymerase expression system. This CRISPR-inspired approach has the potential to expand the application of phages to various basic and translational research projects.

Covalent Modifications of the Bacteriophage Genome Confer a Degree of Resistance to Bacterial CRISPR Systems.

The interplay between defense and counterdefense systems of bacteria and bacteriophages has been driving the evolution of both organisms, leading to their great genetic diversity. Restriction-modification systems are well-studied defense mechanisms of bacteria, while phages have evolved covalent modifications as a counterdefense mechanism to protect their genomes against restriction. Here, we present evidence that these genome modifications might also have been selected to counter, broadly, the CRISPR-Cas systems, an adaptive bacterial defense mechanism. We found that the phage T4 genome modified by cytosine hydroxymethylation and glucosylation (ghmC) exhibits various degrees of resistance to the type V CRISPR-Cas12a system, producing orders of magnitude more progeny than the T4(C) mutant, which contains unmodified cytosines. Furthermore, the progeny accumulated CRISPR escape mutations, allowing rapid evolution of mutant phages under CRISPR pressure. A synergistic effect on phage restriction was observed when two CRISPR-Cas12a complexes were targeted to independent sites on the phage genome, another potential countermechanism by bacteria to more effectively defend themselves against modified phages. These studies suggest that the defense-counterdefense mechanisms exhibited by bacteria and phages, while affording protection against one another, also provide evolutionary benefits for both.IMPORTANCE Restriction-modification (R-M) and CRISPR-Cas systems are two well-known defense mechanisms of bacteria. Both recognize and cleave phage DNA at specific sites while protecting their own genomes. It is well accepted that T4 and other phages have evolved counterdefense mechanisms to protect their genomes from R-M cleavage by covalent modifications, such as the hydroxymethylation and glucosylation of cytosine. However, it is unclear whether such genome modifications also provide broad protection against the CRISPR-Cas systems. Our results suggest that genome modifications indeed afford resistance against CRISPR systems. However, the resistance is not complete, and it is also variable, allowing rapid evolution of mutant phages that escape CRISPR pressure. Bacteria in turn could target more than one site on the phage genome to more effectively restrict the infection of ghmC-modified phage. Such defense-counterdefense strategies seem to confer survival advantages to both the organisms, one of the possible reasons for their great diversity.

Genetic Engineering of Bacteriophages Against Infectious Diseases.

Bacteriophages (phages) are the most abundant and widely distributed organisms on Earth, constituting a virtually unlimited resource to explore the development of biomedical therapies. The therapeutic use of phages to treat bacterial infections ("phage therapy") was conceived by Felix d'Herelle nearly a century ago. However, its power has been realized only recently, largely due to the emergence of multi-antibiotic resistant bacterial pathogens. Progress in technologies, such as high-throughput sequencing, genome editing, and synthetic biology, further opened doors to explore this vast treasure trove. Here, we review some of the emerging themes on the use of phages against infectious diseases. In addition to phage therapy, phages have also been developed as vaccine platforms to deliver antigens as part of virus-like nanoparticles that can stimulate immune responses and prevent pathogen infections. Phage engineering promises to generate phage variants with unique properties for prophylactic and therapeutic applications. These approaches have created momentum to accelerate basic as well as translational phage research and potential development of therapeutics in the near future.

Mitigation of the corrosion-causing Desulfovibrio desulfuricans biofilm using an organic silicon quaternary ammonium salt in alkaline media simulated concrete pore solutions.

Organic silicon quaternary ammonium salt (OSA), an environmentally friendly naturally occurring chemical, was used as a bacteriostatic agent against sulphate-reducing bacteria (SRB) on a 20SiMn steel surface in simulated concrete pore solutions (SCP). Four different media were used: No SRB (NSRB), No SRB and OSA (NSRB + OSA), With SRB (WSRB), With SRB and OSA (WSRB + OSA). After biofilm growth for 28 days, optimized sessile SRB cells survived at the high pH of 11.35 and as a result these cells caused the breakdown of the passive film due to the metabolic activities of the SRB. Corrosion prevention results showed that the OSA was effective in mitigating the growth of the sessile SRB cells and reduced corrosion in the SCP. These results were further confirmed by scanning electron microscope images, energy dispersive X-ray analysis, confocal-laser scanning microscopy, X-ray photoelectron spectroscopy and corrosion testing using electrochemical analysis.

Procalcitonin and C-reactive protein in urinary tract infection diagnosis.

BACKGROUND: Urinary infections are a common type of pediatric disease, and their treatment and prognosis are closely correlated with infection location. Common clinical manifestations and laboratory tests are insufficient to differentiate between acute pyelonephritis and lower urinary tract infection. This study was conducted to explore a diagnostic method for upper and lower urinary tract infection differentiation. METHODS: The diagnostic values of procalcitonin (PCT) and C-reactive protein (CRP) were analyzed using the receiver operating characteristic curve method for upper and lower urinary tract infection differentiation. PCT was determined using chemiluminescent immunoassay. RESULTS: The PCT and CRP values in children with acute pyelonephritis were significantly higher than those in children with lower urinary tract infection (3.90 ± 3.51 ng/ml and 68.17 ± 39.42 mg/l vs. 0.48 ± 0.39 ng/ml and 21.39 ± 14.92 mg/l). The PCT values were correlated with the degree of renal involvement, whereas the CRP values failed to show such a significant correlation. PCT had a sensitivity of 90.47% and a specificity of 88% in predicting nephropathia, whereas CRP had sensitivity of 85.71% and a specificity of 48%. CONCLUSIONS: Both PCT and CRP can be used for upper and lower urinary tract infection differentiation, but PCT has higher sensitivity and specificity in predicting pyelonephritis than CRP. PCT showed better results than CRP. PCT values were also correlated with the degree of renal involvement.

[Expression of microtubule-associated protein 1 light chain 3 in laryngeal carcinoma].

OBJECTIVE: To investigate the expression of microtuble-associated protein 1 light chain 3 (LC3) in laryngeal squamous cell carcinoma (LSCC). METHOD: The expression of LC3 in 50 cases of LSCC, 45 cases of para-carcinoma, 10 cases of laryngeal papilloma and 16 cases of polyp of vocal cord were detected by immunohistochemistry (MaxVision method). Expression level of LC3 mRNA was assayed by RT-PCR in 41 of LSCC, 41 of para-carcinoma tissue and 11 of polyp of vocal cord. RESULT: The positive rates of LC3 protein expression were 60.0%, 93.3%, 90.0%, 93.8% in LSCC tissue, para-carcinoma, laryngeal papilloma and poly of vocal card tissues, respectively. The positive rates of LC3 were significantly lower in LSCC than in para-carcinoma and poly of vocal cord (chi2 = 18.135, P < 0.01). The mRNA levels of LC3 were significantly lower in LSCC than in para-carcinoma and poly of vocal cord (0.57 +/- 0.08 )vs (0.99 +/- 0.11) and (1.07 +/- 0.05) , F = -255.872, P < 0.01. The expression of LC3 were related to tumor location and pathological grade (P < 0.05), but not related to age, T stage, clinical stage and lymphoid metastasis (P > 0.05). CONCLUSION: Expression of LC3 are down-regulated in LSCC. The change of autophagic capacity may play an important role in occurrence and development of LSCC.

Evaluation of right ventricle function in children with primary nephrotic syndrome.

BACKGROUND: We aimed to evaluate right ventricle (RV) function in children with primary nephrotic syndrome (PNS). METHODS: RV hemodynamics were evaluated by Doppler echocardiography in 50 children with PNS (aged 2.5-12 years), either at PNS onset (n = 37) or relapse (n = 13), and in 50 normal controls. Heart rate, stroke volume, cardiac output, RV enddiastolic and end-systolic volume, RV ejection fraction, RV end-diastolic pressure, RV peak systolic and end-systolic pressure were determined from pressure-volume loops. The maximal rates of RV pressure upstroke and fall (dP/d t(max) and dP/d t(min), respectively) were calculated. Effective pulmonary arterial elastance was calculated as end-systolic pressure divided by stroke volume. Plasma tumor necrosis factor-alpha (TNF-alpha) and insulin-like growth factor 1 (IGF-1) were also measured. RESULTS: RV end-diastolic pressure was increased by an average of 20% in 39 of the patients with PNS, whereas RV ejection fraction was reduced by an average of 15% compared with controls (p < 0.05 for both). Cardiac output and stroke volume were maintained, indicating compensation at the expense of increased RV end-diastolic and end-systolic volumes and increased RV filling pressure (p < 0.05). Plasma TNF-alpha was elevated in patients with PNS (326 +/- 117 kU/L vs. 75 +/- 23 kU/L, p < 0.05); IGF-1 was similar in PNS patients and controls. CONCLUSION: Right ventricle function was impaired in children with PNS. The characteristics were unrelated to blood pressure and IGF-1, but may be correlated with TNF-alpha and disease duration. Further studies are needed to evaluate the etiology and clinical implications of this abnormality.

A new experimental method for in situ corrosion monitoring under alternate wet-dry conditions.

A new experimental method was applied in in situ corrosion monitoring of mild steel Q235 under alternate wet-dry conditions. The thickness of the electrolyte film during the wet cycle was monitored by a high-precision balance with a sensibility of 0.1 mg. At the same time, an electrochemical impedance technique was employed to study the effect of film thickness on corrosion rates. Experimental results showed that there was a critical electrolyte film condition for which the corrosion rate reached a maximum during wet-dry cycles. For the substrate, the critical condition could be described by a film thickness of about 17 µm. For the rusted specimen, the critical condition could be described by an electrolyte amount of about 0.038 g, which is equivalent to a film thickness of 38 µm. This monitoring system was very useful for studying atmospheric corrosion of metals covered by corrosion products.

[Computerized tomographic evaluation of traumatic ossicles disruption].

OBJECTIVE: To investigate the importance of the high resolution computerized tomography of temporal bone in evaluation of traumatic lesions, particularly of the ossicular disruption. METHOD: Ten patients with traumatic ossicles dislocation underwent preoperative CT scanning followed by surgical exploration of the middle ear. RESULT: The radiographic finding was consistent with the operative demonstration for malleoincudal disarticulation (3 ears), incudostapedial dislocation (7 ears), and the fracture of temporal bone (8 ears). CONCLUSION: The incus was the most vulnerable ossicle in the trauma to the middle ear. CT scans can diagnose exactly the damage position in middle ear.