Cytokine status indicators in children with acute respiratory viral infections after treatment with intranasal interferon-based medicine

Objective: to study the dynamics of local and systemic cytokine production in children with different clinical forms of acute respiratory viral infections (ARVI), including COVID-19, and to assess the effectiveness of local interferon-based therapy.Patients and methods: The study included 180 patients aged from 1 month to 17 years with сonfirmed acute respiratory viral infections (ARVI), including COVID-19. Patients were divided into 2 groups (main and control) of 90 people each. In the main group patients received the intranasal interferon-based medicine Grippferon® in addition to the basic therapy, the control group patients received only basic therapy. The cytokine status was assessed by the content of IFN-α and -γ, IL-1β, IL-8, IL-4, IL-10, IL-17 in blood serum and in nasopharyngeal secretions by enzyme immunoassay kits ("Cytokine", St. Petersburg).Results: Statistically significant differences were revealed in the systemic and local content of individual cytokines in ARVI of different etiologies, depending on the level of damage to the respiratory tract. The use of the interferon-based medicine Grippferon® for intranasal use in children in the early stages of ARVI, including COVID-19, helps to decrease the high content of cytokines IL-1β and IL-8 in the nasopharynx by reducing the viral load. As a result, the duration of catarrhal disease symptoms and intoxication was also significantly reduced as well as the pathogen elimination time.

Plant Extracts for the Control of Listeria monocytogenes in Meat Products

The antimicrobial activity of garlic (Allium sativum L.) and onion (Allium cepa L.) plant active extracts was determined against Listeria monocytogenes in two meat products. Samples of sausages “cacciatore” and cooked ham in vacuum-packaged slices were artificially contaminated, and the presence of Listeria was evaluated during the sausages ripening and throughout the shelf-life of the cooked ham. The test carried out on sausages did not show differences among treated and untreated samples. The antagonistic activity of the plant extracts against the pathogen was probably hidden by the competition from the sausages microbial flora and the pH and the water activity (aw) decrease. On the other hand, the plant extracts determined an initial reduction of about 1.00 log cfu/g of the L. monocytogenes viable count in the cooked ham slices contaminated with 103 cfu/g, but the best result was obtained with the contamination of 102 cfu/g of L. monocytogenes. In addition to the pathogen’s initial decrease, we observed an extension of the lag phase and a reduction of the Listeria growth rate. Considering that the presence of L. monocytogenes during the slicing phase of the cooked ham does not exceed 10 cfu/g, the use of plant extracts can lead to complete pathogen elimination.

Hybrid Hydrogen Peroxide for Viral Disinfection

Decontamination is often necessary in facilities with sensitive spaces where pathogen elimination is critical. Historically, high concentration vaporized hydrogen peroxide technologies have been applied in these areas for pathogen disinfection. While effective, these high concentration solutions come with inherent risks to human health and safety. Alternatively, one recent innovation is a hybrid hydrogen peroxide system which combines a 7% hydrogen peroxide solution with a calibrated fogging device that delivers a mixture of vaporous and micro aerosolized particles, significantly lowering the risk of exposure to high-concentration hazardous chemicals. Studies performed with this technology demonstrate high level pathogen decontamination across a variety of tested pathogens and substrates. This chapter will cover a brief history of hydrogen peroxide technologies and their application processes; examine the correlations between viral inactivation, viral disinfection, and biological indicators for validation; demonstrate the necessity of dwell time for optimal efficacy; discuss the effects of viral disinfectant use on laboratory surfaces; and examine various studies, including virologic work performed in Biosafety Level 3 facilities and good laboratory practice (GLP) data performed by EPA-approved laboratories. This chapter will provide readers a deeper understanding of essential components and considerations when implementing hydrogen peroxide systems for viral decontamination.

Probiotic-mediated p38 MAPK immune signaling prolongs the survival of Caenorhabditis elegans exposed to pathogenic bacteria

The host-microbiota cross-talk represents an important factor contributing to innate immune response and host resistance during infection. It has been shown that probiotic lactobacilli exhibit the ability to modulate innate immunity and enhance pathogen elimination. Here we showed that heat-inactivated probiotic strain Lactobacillus curvatus BGMK2-41 stimulates immune response and resistance of the Caenorhabditis elegans against Staphylococcus aureus and Pseudomonas aeruginosa. By employing qRT-PCR and western blot analysis we showed that heat-inactivated BGMK2-41 activated PMK-1/p38 MAPK immunity pathway which prolongs the survival of C. elegans exposed to pathogenic bacteria in nematode killing assays. The C. elegans pmk-1 mutant was used to demonstrate a mechanistic basis for the antimicrobial potential of BGMK2-41, showing that BGMK2-41 upregulated PMK-1/p38 MAPK dependent transcription of C-type lectins, lysozymes and tight junction protein CLC-1. Overall, this study suggests that PMK-1/p38 MAPK‐dependent immune regulation by BGMK2-41 is essential for probiotic-mediated C. elegans protection against gram-positive and gram-negative bacteria and could be further explored for development of probiotics with the potential to increase resistance of the host towards pathogens.

A novel pneumococcal protein–polysaccharide conjugate vaccine based on biotin–streptavidin

Pneumococcal disease is a serious public health problem worldwide and an important cause of morbidity and mortality among children and adults in developing countries. Although vaccination is among the most effective approaches to prevent and control pneumococcal diseases, approved vaccines have limited protective effects. We developed a pneumococcal protein–polysaccharide conjugate vaccine that is mediated by the non-covalent interaction between biotin and streptavidin. Biotinylated type IV capsular polysaccharide was incubated with a fusion protein containing core streptavidin and Streptococcus pneumoniae virulence protein and relying on the non-covalent interaction between biotin and streptavidin to prepare the protein–polysaccharide conjugate vaccine. Analysis of vaccine efficacy revealed that mice immunized with the protein–polysaccharide conjugate vaccine produced antibodies with high potency against virulence proteins and polysaccharide antigens and were able to induce Th1 and Th17 responses. The antibodies identified using an opsonophagocytic assay were capable of activating the complement system and promoting pathogen elimination by phagocytes. Additionally, mice immunized with the protein–polysaccharide conjugate vaccine and then infected with a lethal dose of Streptococcus pneumoniae demonstrated induced protective immunity. The data indicated that the pneumococcal protein–polysaccharide (biotin–streptavidin) conjugate vaccine demonstrated broad-spectrum activity applicable to a wide range of people and ease of direct coupling between protein and polysaccharide. These findings provide further evidence for the application of biotin–streptavidin in S. pneumoniae vaccines.

Identification of Pulpitis-Related Potential Biomarkers Using Bioinformatics Approach

Inflammatory reaction of pulp tissue plays a role in the pathogen elimination and tissue repair. The evaluation of severity of pulpitis can serve an instructive function in therapeutic scheme. However, there are many limitations in the traditional evaluation methods for the severity of pulpitis. Based on the Gene Expression Omnibus (GEO) database, our study discovered 843 differentially expressed genes (DEGs) related to pulpitis. Afterwards, we constructed a protein-protein interaction (PPI) network of DEGs and used MCODE plugin to determine the key functional subset. Meanwhile, genes in the key functional subset were subjected to GO and KEGG enrichment analyses. The result showed that genes were mainly enriched in inflammatory reaction-related functions. Next, we screened out intersections of PPI network nodes and pulpitis-related genes. Then, 20 genes were obtained as seed genes. In the PPI network, 50 genes that had the highest correlation with seed genes were screened out using random walk with restart (RWR). Furthermore, 4 pulpitis-related hub genes were obtained from the intersection of the top 50 genes and genes in the key functional subset. Finally, GeneMANIA was utilized to predict genes coexpressed with hub genes, and expression levels of the 4 hub genes in normal and pulpitis groups were analyzed based on GEO data. The result demonstrated that the 4 hub genes were mainly coexpressed with chemokine-related genes and were remarkably upregulated in the pulpitis group. In short, we eventually determined 4 potential biomarkers of pulpitis.

Isolation of Skin Leukocytes Uncovers Phagocyte Inflammatory Responses During Induction and Resolution of Cutaneous Inflammation in Fish

Leukocytes offer a critical layer of protection to the host following skin infections. Delineating the kinetics of cutaneous leukocyte recruitment as well as their anti-microbial and regulatory profiles is challenging since it requires the isolation of adequate cell numbers and maintenance of their functional properties. Herein, we took advantage of a modified procedure to gain insights into the contributions of fish phagocytes through induction and resolution phases of acute cutaneous inflammation in goldfish (Carassius auratus). Our data shows early upregulation of pro-inflammatory cytokines and chemokines, which was paired with neutrophil-dominant leukocyte migration of neutrophils from circulation to the injury site. Recruited neutrophils were associated with high levels of reactive oxygen species (ROS). Following pathogen elimination, a reduction in ROS levels and pro-inflammatory cytokines expression preceded the resolution of inflammation. These results provide a better understanding of the cutaneous immune responses in fish. Moreover, the increased viability and functionality of isolated skin leukocytes opens the door to better understand a range of additional skin diseases.

Immunogenetic Epidemiology of Type 1 Diabetes in 14 Continental Western European Countries

Human leukocyte antigen (HLA) is widely recognized to influence individual Type 1 diabetes (T1D) risk. Here we utilized an immunogenetic epidemiological approach to evaluate the influence of HLA on T1D at the population level. Specifically, we evaluated the correlations between the population frequencies of 127 HLA Class I and II alleles and the population prevalence of T1D in 14 Continental Western European countries to identify a population-level HLA profile for T1D. The results of these analyses generally corroborated prior findings regarding the influence of HLA on T1D risk and protection and revealed several novel HLA-T1D associations. The findings, discussed within the context of the role of HLA in pathogen elimination and autoimmunity, point to a contributory role of exposure to pathogens in the absence of protective HLA in underlying the autoimmune destruction of pancreatic beta cells in T1D.

Antibacterial Agents Adsorbed on Active Carbon: A New Approach for S. aureus and E. coli Pathogen Elimination

Antibiotic overuse and mass production have led to a global problem with the treatment of antibacterial infections. Thus, any possibility to limit the number of antibacterial drugs used will contribute to a decrease in the development of pathogenic bacterial resistance. In this study, the enhanced bacterial growth reduction of pharmaceutical activated carbon (PAC) material with adsorbed antimicrobial agents compared to the activity of pure antibacterial drugs was investigated. Sulfamethoxazole (SMZ) at a concentration of 1.1 mg/mL retained the growth of S. aureus and E. coli at 20.5% and 26.5%, respectively, whereas SMZ adsorbed on PAC increased the reduction of the tested bacteria in the range of 47–72%. The use of PAC with adsorbed gentamycin (G) over 24 h improved the effectiveness of E. coli growth reduction by 50% compared to the application of pure antibiotic (3.6 µg/mL). The increased reduction of S. aureus growth by 6% using G with PAC for a 24-h incubation time compared to the use of pure antibiotics at a concentration of 3.6 µg/mL was observed. The results provide proof-of-principle that the new approach of activated carbon with adsorbed antimicrobial agents could yield an attractive background with potential as a new starting material for S. aureus and E. coli pathogen elimination, e.g., in wound-healing treatment in the future.

Targeting Cytokines, Pathogen-Associated Molecular Patterns, and Damage-Associated Molecular Patterns in Sepsis via Blood Purification

Sepsis is characterized by a dysregulated immune response to infections that causes life-threatening organ dysfunction and even death. When infections occur, bacterial cell wall components (endotoxin or lipopolysaccharide), known as pathogen-associated molecular patterns, bind to pattern recognition receptors, such as toll-like receptors, to initiate an inflammatory response for pathogen elimination. However, strong activation of the immune system leads to cellular dysfunction and ultimately organ failure. Damage-associated molecular patterns (DAMPs), which are released by injured host cells, are well-recognized triggers that result in the elevation of inflammatory cytokine levels. A cytokine storm is thus amplified and sustained in this vicious cycle. Interestingly, during sepsis, neutrophils transition from powerful antimicrobial protectors into dangerous mediators of tissue injury and organ dysfunction. Thus, the concept of blood purification has evolved to include inflammatory cells and mediators. In this review, we summarize recent advances in knowledge regarding the role of lipopolysaccharides, cytokines, DAMPs, and neutrophils in the pathogenesis of sepsis. Additionally, we discuss the potential of blood purification, especially the adsorption technology, for removing immune cells and molecular mediators, thereby serving as a therapeutic strategy against sepsis. Finally, we describe the concept of our immune-modulating blood purification system.