Elimination Of Pathogens Research Articles

The Src-family Kinase Lyn in Immunoreceptor Signaling.

Effective regulation of immune-cell activation is critical for ensuring that the immune response, and inflammation generated for the purpose of pathogen elimination, are limited in space and time to minimize tissue damage. Autoimmune disease can occur when immunoreceptor signaling is dysregulated, leading to unrestrained inflammation and organ damage. Conversely, tumors can coopt the tissue healing and immunosuppressive functions of hematopoietic cells to promote metastasis and evade therapy. The Src-family kinase Lyn is an essential regulator of immunoreceptor signaling, initiating both proinflammatory and suppressive signaling pathways in myeloid immune cells (eg, neutrophils, dendritic cells, monocytes, macrophages) and in B lymphocytes. Defects in Lyn signaling are implicated in autoimmune disease, but mechanisms by which Lyn, expressed along with a battery of other Src-family kinases, may uniquely direct both positive and negative signaling remain incompletely defined. This review describes our current understanding of the activating and inhibitory contributions of Lyn to immunoreceptor signaling and how these processes contribute to myeloid and B-cell function. We also highlight recent work suggesting that the 2 proteins generated by alternative splicing of lyn, LynA and LynB, differentially regulate both immune and cancer-cell signaling. These principles may also extend to other Lyn-expressing cells, such as neuronal and endocrine cells. Unraveling the common and cell-specific aspects of Lyn function could lead to new approaches to therapeutically target dysregulated pathways in pathologies ranging from autoimmune and neurogenerative disease to cancer.

Evolution of Toll, Spatzle and MyD88 in insects: the problem of the Diptera bias

BackgroundArthropoda, the most numerous and diverse metazoan phylum, has species in many habitats where they encounter various microorganisms and, as a result, mechanisms for pathogen recognition and elimination have evolved. The Toll pathway, involved in the innate immune system, was first described as part of the developmental pathway for dorsal-ventral differentiation in Drosophila. Its later discovery in vertebrates suggested that this system was extremely conserved. However, there is variation in presence/absence, copy number and sequence divergence in various genes along the pathway. As most studies have only focused on Diptera, for a comprehensive and accurate homology-based approach it is important to understand gene function in a number of different species and, in a group as diverse as insects, the use of species belonging to different taxonomic groups is essential.ResultsWe evaluated the diversity of Toll pathway gene families in 39 Arthropod genomes, encompassing 13 different Insect Orders. Through computational methods, we shed some light into the evolution and functional annotation of protein families involved in the Toll pathway innate immune response. Our data indicates that: 1) intracellular proteins of the Toll pathway show mostly species-specific expansions; 2) the different Toll subfamilies seem to have distinct evolutionary backgrounds; 3) patterns of gene expansion observed in the Toll phylogenetic tree indicate that homology based methods of functional inference might not be accurate for some subfamilies; 4) Spatzle subfamilies are highly divergent and also pose a problem for homology based inference; 5) Spatzle subfamilies should not be analyzed together in the same phylogenetic framework; 6) network analyses seem to be a good first step in inferring functional groups in these cases. We specifically show that understanding Drosophila’s Toll functions might not indicate the same function in other species.ConclusionsOur results show the importance of using species representing the different orders to better understand insect gene content, origin and evolution. More specifically, in intracellular Toll pathway gene families the presence of orthologues has important implications for homology based functional inference. Also, the different evolutionary backgrounds of Toll gene subfamilies should be taken into consideration when functional studies are performed, especially for TOLL9, TOLL, TOLL2_7, and the new TOLL10 clade. The presence of Diptera specific clades or the ones lacking Diptera species show the importance of overcoming the Diptera bias when performing functional characterization of Toll pathways.

The Complement System: A Powerful Modulator and Effector of Astrocyte Function in the Healthy and Diseased Central Nervous System.

The complement system, an effector arm of the innate immune system that plays a critical role in tissue inflammation, the elimination of pathogens and the clearance of dead cells and cell debris, has emerged as a regulator of many processes in the central nervous system, including neural cell genesis and migration, control of synapse number and function, and modulation of glial cell responses. Complement dysfunction has also been put forward as a major contributor to neurological disease. Astrocytes are neuroectoderm-derived glial cells that maintain water and ionic homeostasis, and control cerebral blood flow and multiple aspects of neuronal functioning. By virtue of their expression of soluble as well as membrane-bound complement proteins and receptors, astrocytes are able to both send and receive complement-related signals. Here we review the current understanding of the multiple functions of the complement system in the central nervous system as they pertain to the modulation of astrocyte activity, and how astrocytes use the complement system to affect their environment in the healthy brain and in the context of neurological disease.

CARD9 Deficiency Increases Hippocampal Injury Following Acute Neurotropic Picornavirus Infection but Does Not Affect Pathogen Elimination

Neurotropic viruses target the brain and contribute to neurologic diseases. Caspase recruitment domain containing family member 9 (CARD9) controls protective immunity in a variety of infectious disorders. To investigate the effect of CARD9 in neurotropic virus infection, CARD9−/− and corresponding C57BL/6 wild-type control mice were infected with Theiler’s murine encephalomyelitis virus (TMEV). Brain tissue was analyzed by histology, immunohistochemistry and molecular analyses, and spleens by flow cytometry. To determine the impact of CARD9 deficiency on T cell responses in vitro, antigen presentation assays were utilized. Genetic ablation of CARD9 enhanced early pro-inflammatory cytokine responses and accelerated infiltration of T and B cells in the brain, together with a transient increase in TMEV-infected cells in the hippocampus. CARD9−/− mice showed an increased loss of neuronal nuclear protein+ mature neurons and doublecortin+ neuronal precursor cells and an increase in β-amyloid precursor protein+ damaged axons in the hippocampus. No effect of CARD9 deficiency was found on the initiation of CD8+ T cell responses by flow cytometry and co-culture experiments using virus-exposed dendritic cells or microglia-enriched glial cell mixtures, respectively. The present study indicates that CARD9 is dispensable for the initiation of early antiviral responses and TMEV elimination but may contribute to the modulation of neuroinflammation, thereby reducing hippocampal injury following neurotropic virus infection.

Characterizing the Mechanism of NE and P3 Cross-Presentation in B Cell-Acute Lymphoblastic Leukemia

IntroductionPR1 is a nonameric HLA-A2 restricted leukemia-associated antigen derived from serine proteases neutrophil elastase (NE) and proteinase 3 (P3). NE and P3 are primarily expressed in cells of myeloid lineage including granulocytes, bone marrow progenitors and myeloid leukemia. However, NE and P3 are also cross-presented by antigen presenting cells (APCs) such as B-cells and some non-hematopoietic solid tumors, a mechanism whereby exogenous antigens are endocytosed and presented on HLA class I molecules. Therefore, identifying non-myeloid tumors capable of PR1 cross-presentation broadens the application of PR1-targeted immunotherapies, which to date include PR1 peptide vaccine, PR1 cellular therapy, and 8F4, a T cell receptor (TCR)-like monoclonal antibody (mAb).One possible source of NE and P3 in the microenvironment is neutrophil extracellular traps (NETs). NETs are composed of deoxyribonucleic acid (DNA)/histones and the antimicrobial proteases, NE and P3, which are extruded from activated polymorphonuclear neutrophils (PMNs) during inflammation. Although a main function of NETs is elimination of pathogens, studies have shown immune-regulatory effects of NETs. Thus there is a great interest in understanding the role of NETs in modulating adaptive immune system in the setting of anti-tumor immunity. We hypothesized that NETs facilitate NE and P3 cross-presentation in malignant B-cell acute lymphoblastic leukemia (ALL) due to their high abundance in the bone marrow, which could lead to susceptibility to PR1-targeted immunotherapies.MethodsB-ALL cell lines SB, Nalm6, RS4 and Nalm6 were co-cultured with soluble NE and P3 (10 ug/mL) and PMNs at a ratio of 3:1 (PMN: ALL) in low serum media and analyzed for NE and P3 cross-presentation by staining for HLA-A2/PR1 complex using 8F4 mAb. T2 cells pulsed with PR1 and HIV peptide were used as a positive and negative control, respectively. To determine susceptibility to killing my PR1-CTLs, SB cell line was co-cultured with 10 μg/mL soluble NE and P3 for 24 hours and incubated with healthy PR1-CTLs for 4 hours following a protocol for a standard calcein-AM cytotoxicity assay. T2 cells were pulsed with PR1 and CG1 peptide as a positive and negative control, respectively.To identify NETs as a source of NE and P3, healthy PMNs were treated with ionomycin to induce NETosis, co-cultured with B-ALL cell lines at a ratio of 3:1 (PMN: ALL) overnight and analyzed for NE and P3 uptake. NETs were also treated with DNAse to determine the effect on uptake.ResultsThe B-ALL cell line, SB, cross-presented PR1 from soluble P3 and NE, as well as supernatants derived from quiescent PMNs. There was a 1.3 fold significant increase in PR1/HLA-A2 surface expression when co-cultured with soluble and PMN-associated protease (Figure 1A). We then investigated whether PR1 cross-presentation by SB cell line rendered it susceptible to killing by PR1-CTLs. Dose-dependent killing of SB cells was observed following PR1 cross-presentation compared to nonpulsed cells (Figure 1B). The percent specific lysis was evident even at a very low ratio of 0.625:1 Effector: Target cells (E:T). As controls, T2 cells pulsed with PR1 showed dose-dependent killing by PR1-CTLs, whereas T2 cells pulsed with CG1 and nonpulsed did not, as expected (Figure 1B).To evaluate whether NET-associated NE and P3 facilitate uptake in B-ALL, healthy PMNs were co-cultured with B-ALL cell lines overnight. The co-culture resulted in both NE (Figure 2A-B) and P3 (Figure 2C-D) uptake in RS4 and Nalm6 cell lines with an average MFI of 350 and 426, respectably, compared to nonpulsed MFI of 145. Measurement of intracellular P3 and NE by flow cytometry showed that although both NET-associated and PMN-associated NE and P3 are taken up, uptake of supernatants from resting PMNs was significantly greater than NET-associated (Figure 2). Similar to uptake of soluble and PMN-associated proteases, uptake between B-ALL cell lines was heterogeneous.ConclusionCollectively, our data demonstrate that B-ALL cell lines are capable of PR1 cross-presentation rendering them susceptible to killing by PR1-CTLs. Further, NET-associated NE and P3 are a source of protease uptake. Because NET-associated antigens are less efficient at uptake, our data suggest that NETs produced during systemic inflammation might be less immunogenic in B-cell malignancies. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Поліпшення ефективності лікування інфекції сечовивідних шляхів у вагітних

The aim is to evaluate the effectiveness of Phytolysin Active prescription in therapy of urinary tract infections in pregnant women. Materials and methods. The efficacy of Phytolysin Active (1 capsule per day for 21 days) in the complex treatment for urinary tract infection was studied in comparison with the group of pregnant women using traditional antimicrobial therapy. The first group consisted of 27 pregnant women who received complex treatment (antibiotic therapy + Phytolysin Active), the second consisted of 25 women who used exclusively antibiotic therapy. Among pregnant women in the first group, symptomatic gestational cystitis was observed in 9 (33.3%) cases, and in the second group in 8 (32%) patients. Results. The effectiveness of combination therapy with the use of Phytolysin Active was proved by significant improvement in general well-being and disappearance of cystitis symptoms in 8 (88.9%) women compared with 5 (62.5%) patients in the treatment group who used exclusively antimicrobial drugs. After treatment with the addition of Phytolysin Active to antibiotic therapy, Escherichiaсoli pathogen, which is most often diagnosed in this pathology, was observed in 1 (3.7%) women compared to 5 (20%) cases in the second group. Also, 25 (92.6%) women of the first group had no recurrence of urinary tract infections for 3 months. Conclusions. Complex treatment of asymptomatic bacteriuria in pregnant women with the use of the drug Phytolysin Active is more effective than the use of antibiotic therapy alone, which is proved by the high rate of the pathogen elimination and absence of infection recurrence for 3 months. The synergistic action of two active components of Phytolysin Active (Polpharma) – cranberry proanthocyanidins and lactobacillus acidophilus – improves the effectiveness of treatment and prevents relapse. The study was carried out in accordance with the principles of the Declaration of Helsinki. The study protocol was approved by the Local Ethics Committee of the institution. The informed consent of women was obtained for the research. The authors declare no conflicts of interest. Key words: pregnancy, urinary tract infections, treatments, herbal medicine, cranberry, lactobacilli.

Improvement of Must Fermentation from Late Harvest cv. Tempranillo Grapes Treated with Pulsed Light.

Pulsed light irradiation is a nonthermal technology currently used for the elimination of pathogens from a diverse range of food products. In the last two decades, the results obtained using PL at laboratory scale are encouraging wine experts to use it in the winemaking industry. PL can reduce native yeast counts significantly, which facilitates the use of starter cultures, reducing SO2 requirements at the same time. In this experimental set up, Tempranillo grapes were subjected to pulsed light treatment, and the fermentative performance of non-Saccharomyces yeasts belonging to the species Schizosaccharomyces pombe, Lachancea thermotolerans, Torulaspora delbrueckii, Metschnikowia pulcherrima and Hanseniaspora vineae was monitored in sequential fermentations against spontaneous fermentation and pure culture fermentation with the species Saccharomyces cerevisiae. The experimental analyses comprised the determination of anthocyanin (High performance liquid chromatography with photodiode array detector—HPLC-DAD), polyphenol index and colour (Ultraviolet-visible spectroscopy—UV-Vis spectrophotometer), fermentation-derived volatiles (Gas chromatography with flame ionization detector—GC-FID), oenological parameters (Fourier transform Infrared spectroscopy—FT-IR) and structural damage of the skin (atomic force microscopy—AFM). The results showed a decrease of 1.2 log CFU/mL yeast counts after pulsed light treatment and more rapid and controlled fermentation kinetics in musts from treated grapes than in untreated samples. The fermentations done with treated grapes allowed starter cultures to better implant in the must, although a larger anthocyanin loss (up to 93%) and an increase in hue values (1 unit) towards more yellow hues were observed for treated grapes. The development of biomass was larger in musts from treated grapes. The profile of volatile compounds and oenological parameters reveals that fermentations carried out with untreated grapes are prone to deviations from native microbiota (e.g., production of lactic acid). Finally, no severe damage on the skin was observed with the AFM on treated grapes.

MO669FIRST RESULTS OF THE COVID-19 PATIENTS TREATED WITH THE SERAPH® 100 MICROBIND® AFFINITY FILTER (COSA) REGISTRY

Background and AimsThe COVID-19 pandemic has serious impact on health and economics worldwide. Despite the recent advent of SARS-Cov-2 vaccines, treatment options are needed, yet pharmacologic interventions remain limited. Several extracorporeal treatments are currently explored concerning their potential to improve the clinical course and outcome of critically ill patients with COVID-19. The Seraph® 100 Microbind® Affinity adsorber (Exthera Medical, CA, USA) has recently been introduced for the elimination of several pathogens from the blood and an emergency authorization in patients with COVID-19 was granted by the FDA. Bacteria, viruses (including the SARS-CoV-2 spike glycoprotein), fungi and toxins have been shown to bind to the immobilized heparin on the ultra-high molecular weight polyethylene beads of the device in a similar way to the interaction with heparan sulfate on the cell-surface and are thereby removed from the bloodstream. Here we report the interim analysis of the COVID-19 patients treated with the Seraph® 100 Microbind® Affinity filter (COSA) registry. The goal of the registry is to gather data regarding the safety and efficacy of the Seraph® 100 in the treatment of COVID-19 patients.MethodParticipating sites were advised to insert patient data of COVID-19 patients, treated with the Seraph® 100, during their hospital stay into the COSA registry (ClinicalTrials.gov Identifier: NCT04361500). A total of 66 items were asked in a web-based platform.ResultsUntil January 2021, 33 patients with 39 treatment sessions form six different hospitals were reported to the register (seven female, median age 61 years, Table 1). The patients were treated between March and December 2020. Eleven patients with a hospital admission between March and August and 22 between September and December 2020. The Seraph® 100 treatment was initiated 9 days after symptom onset, without any difference between survivors and non-survivors. Overall, a mortality of 27% was reported. Serious comorbidities (as preadmission immunosuppressive therapy, lung fibrosis or CKD5T) were reported in all of the non-survivors. Invasive ventilation was needed in 67% of these patients when Seraph treatment was initiated. A non-significant trend towards higher Ferritin levels in non-survivors (2000 (1963 – 8326) vs. 989 (644 – 2000), p=0.09) was reported. All treatments were well tolerated, three clotting events were reported.ConclusionViral SARS-CoV-2 RNA is frequently (up to 78%) seen in the blood of critically ill COVID-19 patients and correlates with the severity of the disease. The Seraph® 100 can bind to viral spike protein, proinflammatory cytokines may be reduced by the device and hemodynamic stabilization has been reported during the Seraph®100 treatment of COVID-19 patients. Platelets can be hyperactivated in association with SARS-CoV-2 proteins and thus presumably trigger the hypercoagulation and thrombosis. In this context, the removal of SARS-CoV-2 proteins to prevent hyperactivated platelets appears to be a sensible approach.All reported deaths were associated with serious preexisting comorbidities, immunosuppression, dialysis dependent renal failure, or a combination of these factors. Hence, Seraph® 100 treatment may be most beneficial in COVID-19 courses of patients without multi organ failure. More clinical data is needed to describe possible benefits of the Seraph®100 in the treatment of COVID-19 patients.

Effect of probiotic Lactobacillus rhamnosus by-products on gingival epithelial cells challenged with Porphyromonas gingivalis

ObjectiveProbiotics are usually given as living cells, but their effects may be also achieved by postbiotics. We hypothesized that probiotics products (spent media and lysate) altered the response induced by P. gingivalis in gingival epithelial cells (GECS). MethodsImmortalized human OBA-9 GECs (∼2,5 × 105cells/well) were challenged with P. gingivalis ATCC33277, and co-infected with L. rhamnosus Lr-32 for 4 h. L. rhamnosus Lr-32 spent medium or cells lysate was added to GECs co-infected with P. gingivalis. Another set of OBA-9 GECs were first exposed to P. gingivalis ATCC 33277 and then to the living probiotic or probiotic products. Transcription of genes encoding inflammatory mediators (IL-1β, TNF-α, IL-6, and CXCL-8) and receptors (TLR2 and TLR4) were evaluated by RT-qPCR. P. gingivalis growth under L. rhamnosus Lr-32 postbiotics was also evaluated. ResultsL. rhamnosus Lr-32 spent media decreased cell viability, while living cells and cell lysates did not. L. rhamnosus Lr-32 lysate, but not spent media, upregulated transcription of inflammatory mediators (IL-1β, TNF-α, IL-6, and CXCL-8) in GECs infected with P. gingivalis. Transcription of TRL2 was upregulated in all experimental groups compared to control, whereas TLR4 was upregulated by the probiotic or its postbiotics in P. gingivalis infected cells. Spent media and lysates reduced the growth of P. gingivalis. ConclusionL. rhamnosus Lr-32 cell components rather than live probiotic enhanced the expression of inflammatory mediators in P. gingivalis infected gingival epithelial cells. The increased potential of Lr-32 cell lysates to promote immune response to the periodontopathogen may favor pathogen elimination but may also lead to additional deleterious effects of the exacerbated inflammation.

Health Monitoring of Laboratory Rodent Colonies-Talking about (R)evolution.

Simple SummaryWhen working with laboratory animals, scientists need to ensure that animals are not suffering from natural infections. Therefore, hygienic standards have been developed over the last 100+ years. The key element of hygienic standardization is the monitoring of infectious agents that can compromise the animal’s health, are dangerous for the personnel or interfere with the research. However, scientists became aware that by eliminating such unwanted infectious agents the overall diversity of all microbes in research animals, the so-called microbiome, has also been reduced. Moreover, it became clear that the microbiome composition has an enormous impact on how research models react, e.g., to treatments. This might hinder the translation of findings in preclinical research to the clinical situation. Therefore, new concepts in hygienic standardization need to be developed that take animal welfare as well as scientific value into account. In this review, we will give an overview of how classical monitoring has been evolved and demonstrate concepts regarding how to handle the microbiome as a yet unknown variable in animal-based research in order to enhance the validity of research findings.The health monitoring of laboratory rodents is essential for ensuring animal health and standardization in biomedical research. Progress in housing, gnotobiotic derivation, and hygienic monitoring programs led to enormous improvement of the microbiological quality of laboratory animals. While traditional health monitoring and pathogen detection methods still serve as powerful tools for the diagnostics of common animal diseases, molecular methods develop rapidly and not only improve test sensitivities but also allow high throughput analyses of various sample types. Concurrently, to the progress in pathogen detection and elimination, the research community becomes increasingly aware of the striking influence of microbiome compositions in laboratory animals, affecting disease phenotypes and the scientific value of research data. As repeated re-derivation cycles and strict barrier husbandry of laboratory rodents resulted in a limited diversity of the animals’ gut microbiome, future monitoring approaches will have to reform—aiming at enhancing the validity of animal experiments. This review will recapitulate common health monitoring concepts and, moreover, outline strategies and measures on coping with microbiome variation in order to increase reproducibility, replicability and generalizability.

A C-Type Lectin Highly Expressed in Portunus trituberculatus Intestine Functions in AMP Regulation and Prophenoloxidase Activation.

A C-type lectin (PtCLec2) from Portunus trituberculatus was identified for characterization of its role in defense and innate immunity. PtCLec2 contains a single carbohydrate-recognition domain (CRD) with a conserved QPD motif, which was predicted to have galactose specificity. The mRNA expression of PtCLec2 was predominantly detected in intestine and increased rapidly and significantly upon pathogen challenge. The recombinant PtCLec2 (rPtCLec2) could bind various microorganisms and PAMPs with weak binding ability to yeast and PGN. It agglutinated the tested Gram-negative bacteria (Vibrio alginolyticus and Pseudomonas aeruginosa), Gram-positive bacteria (Staphylococcus aureus and Micrococcus luteus), and rabbit erythrocytes in the presence of exogenous Ca2+, and these agglutination activities were suppressed by LPS, d-galactose, and d-mannose. Further, rPtCLec2 enhanced phagocytosis and clearance of V. alginolyticus, and displayed inhibitory activities against the tested bacteria. Knockdown of PtCLec2 decreased the transcription of two phagocytosis genes (PtArp and PtMyosin), three prophenoloxidase (proPO) system-related genes (PtPPAF, PtcSP1, and PtproPO), six antimicrobial peptides (AMPs) (PtALF4-7, PtCrustin1, and PtCrustin3), and PtRelish but upregulated the expression levels of PtJNK, PtPelle, and PtTLR. These results collectively indicate that PtCLec2 might perform its immune recognition function via binding and agglutination, and mediate pathogen elimination via regulating hemocyte phagocytosis, AMP synthesis, and proPO activation.

Daily intake of a homeopathic agent by dogs modulates white cell defenses and reduces bacterial counts in feces

Motivated by conventional medicines' ineffectiveness and the appearance of bacterial resistance, homeopathic medicines are being increasingly demanded by the veterinary market. Dog feces contain high bacterial and pathogenic loads, which can cause diseases in animals and facilitate zoonotic transmission. Adequate animal immunity contributes to the elimination of potential pathogens. This work's objective was to evaluate the effects of using a homeopathic medication on blood cell counts, serum protein levels, and fecal bacterial counts in dogs. Two groups were used, each including five 19-month-old Beagle dogs. The homeopathic product Orgainfecto® was sprayed on the two daily meals in the treated group (T), at 0.5 mL/animal/day for 60 days; the group that did not receive the homeopathic was considered the control (C), who were fed only the vehicle used in the production of the homeopathic as a placebo. On days 1, 30, and 60, individual stool samples were collected for bacterial counts in the stool, blood samples for blood counts, and serum for biochemical tests. Total bacterial counts in feces were lower in group T on days 30 and 60 of the experiment, while total coliforms and E coli. were lower in this group on day 60. Total leukocyte counts were lower on day 60 in group T, unlike the neutrophil count, higher in group T on day 30, and eosinophils on day 60. However, the lymphocyte count was lower in group T on days 30 and 60. There was an increase in globulin and total protein in group T on day 60. We conclude that the tested homeopathic causes neutrophils and eosinophils' stimulation and increases globulins levels even with reduced lymphocytes. The daily consumption of homeopathic by dogs decreases the intestinal bacterial load, resulting in less environmental contamination and animal and human colibacillosis risks.

Optimising pyrolysis conditions for high-quality biochar production using black soldier fly larvae faecal-derived residue as feedstock

The disposal of feacal matter from Urine Diversion Dry Toilets is a significant challenge due to limited land availability, possible underground water contamination, and the risk of spreading diseases. The collected faecal matter can be fed to Black Soldier Fly Larvae to produce protein-rich larvae used as animal feed. The disposal of the leftover waste (BSFL residue) is still a problem due to the risk of residual pathogen contamination. The BSFL residue contains residual plant nutrients and can be further processed into biochar. Faecal matter biochar offers an exciting value proposition where the pyrolysis process guarantees a 100% pathogen elimination. It also results in significant waste reduction in transport, storage weight, and volume. A preliminary study was conducted to (i) optimise pyrolysis conditions (optimal temperature treatment and residence time) for biochar production using residue obtained after faecal matter from urine diversion dry toilets was fed to black soldier fly larvae as feedstock; and (ii) determine the physicochemical and morphological characteristics of biochar produced. The residue was pyrolysed at 300, 400, and 500 °C and characterised for chemical, biological and physical characteristics. Surface area (6.61 m2 g−1), pore size, and C: N (9.28) ratio increased at 500 °C for 30 min. Exchangeable bases, (Calcium) Ca, (Magnesium) Mg, (Potassium) K, and (Sodium) Na increased with increasing pyrolysis temperature. The increase in basic cations resulted in an increase in pH from 6.7 in the residue to 9.8 in biochar pyrolysed at 500 °C. Biochar pyrolysed at 500 °C can therefore be used to improve acidic soils. Phosphorus increased with increasing pyrolysis temperature to 3 148 mg kg−1 at 500 °C. Biochar produced at 500 °C for 30 min had desirable characteristics: surface area, exchangeable bases, and pH. Also, biochar can be used as a phosphorus source with potential for crop production, although an external nitrogen source is needed to meet crop nutrient requirements.

Gold Nanomaterials as a Promising Integrated Tool for Diagnosis and Treatment of Pathogenic Infections—A Review

This review summarizes research on functionalized gold nanomaterials as pathogen detection sensors and pathogen elimination integrated tools. After presenting the challenge of current severe threat from pathogenic bacteria and the increasingly serious growth rate of drug resistance, the first section mainly introduces the conspectus of gold nanostructures from synthesis, characterization, physicochemical properties and applications of gold nanomaterials. The next section deals with gold nanomaterials-based pathogen detection sensors such as colorimetric sensors, fluorescence sensors and Surface-Enhanced Raman Scattering sensors. We then discuss strategies based on gold nanomaterials for eliminating pathogenic infections, such as the dual sterilization strategy for grafting gold nanomaterials with antibacterial substances, photothermal antibacterial and photodynamic antibacterial methods. The fourth part briefly introduces the comprehensive strategy for diagnosis and sterilization of pathogen infection based on gold nanomaterials, such as the diagnosis and treatment strategy for pathogen infection using Roman signals real-time monitoring and photothermal sterilization. A concluding section that summarizes the current status and challenges of the novel diagnosis and treatment integrated strategy for pathogenic infections, gives an outlook on potential future perspectives.

Neutrophil subtypes shape HIV-specific CD8 T-cell responses after vaccinia virus infection

Neutrophils are innate immune cells involved in the elimination of pathogens and can also induce adaptive immune responses. Nα and Nβ neutrophils have been described with distinct in vitro capacity to generate antigen-specific CD8 T-cell responses. However, how these cell types exert their role in vivo and how manipulation of Nβ/Nα ratio influences vaccine-mediated immune responses are not known. In this study, we find that these neutrophil subtypes show distinct migratory and motility patterns and different ability to interact with CD8 T cells in the spleen following vaccinia virus (VACV) infection. Moreover, after analysis of adhesion, inflammatory, and migration markers, we observe that Nβ neutrophils overexpress the α4β1 integrin compared to Nα. Finally, by inhibiting α4β1 integrin, we increase the Nβ/Nα ratio and enhance CD8 T-cell responses to HIV VACV-delivered antigens. These findings provide significant advancements in the comprehension of neutrophil-based control of adaptive immune system and their relevance in vaccine design.

Molecular Mechanisms Responsible for Mesenchymal Stem Cell-Based Treatment of Viral Diseases.

Mesenchymal stem cells (MSCs) are adult, immunomodulatory stem cells which reside in almost all postnatal tissues. Viral antigens and damage-associated molecular patterns released from injured and infected cells activate MSCs, which elicit strong antiviral immune response. MSC-sourced interferons and inflammatory cytokines modulate the cytotoxicity of NK cells and CTLs, enhance the antigen-presentation properties of DCs and macrophages, regulate cytokine synthesis in CD4+ T helper cells and promote antibody production in B cells. After the elimination of viral pathogens, MSCs produce immunoregulatory cytokines and trophic factors, prevent the over-activation of immune cells and promote tissue repair and regeneration. In this review article, we summarize the current knowledge on the molecular mechanisms that are responsible for the MSC-dependent elimination of virus-infected cells, and we emphasize the therapeutic potential of MSCs and their secretomes in the treatment of viral diseases.

Minimising the risk of cross infection between siblings with cystic fibrosis (CF) within the home: Successful domestic steam disinfection of CF bacterial and foodborne pathogens on common household cutlery and crockery utensils

Avoidance of cross infection is important between CF siblings living in the same household. In this study, we examined the ability of domestic steam disinfection to eradicate 16 species of CF bacterial and foodborne pathogens from the surface of contaminated crockery and cutlery. Domestic steam disinfection employing baby bottle disinfector devices, when performed properly under manufacturer's instructions, eradicated all organisms tested and offers a relatively inexpensive, simple, versatile and widely available technology for the elimination of common CF bacterial and foodborne pathogens from contaminated crockery and cutlery utensils. Most CF households may already have a baby bottle disinfector device, in order to disinfect nebulizers, respiratory equipment and toothbrushes and therefore employment of this device to disinfect crockery and cutlery would be easily achieved. We therefore advocate the employment of such devices to disinfect such common household utensils, as a critical control in the elimination of these organisms from these sources, thereby enhancing CF sibling safety.

An overview of multifaceted role of Trichoderma spp. for sustainable agriculture

The excessive use of chemical fertilizers and pesticides have caused several negative impacts on the environment and human health. They degrade soil fertility, build up resistance on pathogens, inhibit microbial activities and also enhance greenhouse gas emission. It is impossible and inappropriate to control plant pathogens by using chemical pesticides alone. Emphasize should be given towards organic fertilizers and pesticides to attain sustainability in agriculture. The use of Trichoderma is slowly increasing in the recent years among progressive farmers as an alternative to chemical fertilizers and pesticides. Slow rate of multiplication and colonization, susceptible to biotic and abiotic stresses, incomplete elimination of pathogens and high cost are the major problems behind its poor adoption among the farmers. To overcome these challenges different strains of Trichoderma should be identified which can multiply and colonize rapidly, least affected by environmental conditions and having wide host range on pathogens. In addition, farmers should be made aware about the importance of Trichoderma in agriculture through various extension facilities for its wide scale adoption. Trichoderma can be the viable and sustainable alternative which acts as biofertilizer, bioremediator and biocontrol agent. Nevertheless, the use of Trichoderma is limited on research activities and its application at farmers' level is not yet satisfactory. Thus, this study based on critical analysis of the research works from worldwide researchers aims to reveal the present scenario of the use of Trichoderma, its importance, modes of action, methods of application and multiplication, challenges for wide scale adoption and its appropriate solutions.

Back to basics in COVID-19: Antigens and antibodies-Completing the puzzle.

The outbreak of the coronavirus disease 2019 (COVID‐19) has gathered 1 year of scientific/clinical information. This informational asset should be thoroughly and wisely used in the coming year colliding in a global task force to control this infection. Epidemiology of this infection shows that the available estimates of SARS‐CoV‐2 infection prevalence largely depended on the availability of molecular testing and the extent of tested population. Within molecular diagnosis, the viability and infectiousness of the virus in the tested samples should be further investigated. Moreover, SARS‐CoV‐2 has a genetic normal evolution that is a dynamic process. The immune system participates to the counterattack of the viral infection by pathogen elimination, cellular homoeostasis, tissue repair and generation of memory cells that would be reactivated upon a second encounter with the same virus. In all these stages, we still have knowledge to be gathered regarding antibody persistence, protective effects and immunological memory. Moreover, information regarding the intense pro‐inflammatory action in severe cases still lacks and this is important in stratifying patients for difficult to treat cases. Without being exhaustive, the review will cover these important issues to be acknowledged to further advance in the battle against the current pandemia.

Leishmania (V.) braziliensis infection promotes macrophage autophagy by a LC3B-dependent and BECLIN1-independent mechanism

Leishmania (Viannia) braziliensis is one of the main etiological agents of tegumentary leishmaniasis in Latin America. The establishment of a successful infection in host cells requires several key events including phagocytosis, phagolysosomal maturation impairment, and parasite replication. Autophagy is accountable for the physiological turnover of cellular organelles, degradation of macromolecular structures, and pathogen elimination. In many cases, autophagy control leads to a successful infection, both impairing pathogen elimination or providing nutrients. Here, we have investigated the relationship between autophagy and L. braziliensis infection. We observed that BECLIN1 expression was upregulated early on infection in both in vitro macrophage cultures and biopsies of cutaneous lesions from L. braziliensis infected patients. On the other hand, LC3B expression was downregulated in cutaneous lesions biopsies. A transient pattern of LC3+ cells was observed along L. braziliensis infection, but the number of LC3 puncta did not vary. Additionally, autophagy induction, with rapamycin treatment or through starvation, reduced infection. As expected, rapamycin increased the percentage of LC3+ cells and the number of puncta, but the presence of parasite restricted this effect, indicating LC3-associated autophagy impairment by L. braziliensis. Finally, silencing LC3B but not BECLIN1 promoted infection, confirming BECLIN1 independent and LC3B-related control by the parasite. Taken together, these data indicate macrophage autophagic machinery manipulation by L. braziliensis, resulting in successful establishment and survival into the host cell.