Study on the mechanism of Yupingfeng powder in the treatment of immunosuppression based on UPLC⁃QTOF⁃MS, network pharmacology and molecular biology verification.

AIMS: The current study aims to investigate the effect of Yupingfeng (YPF) powder on immunosuppression, and explore the possible mechanisms. MAIN METHODS: Firstly, the monomer components of YPF powder were analyzed by UPLC-QTOF-MS combined with UNIFI automatic analysis platform, then the mechanism of YPF on immunosuppressive treatment was investigated using network pharmacological method, and finally the prediction was verified in a Candida albicans (Can)-induced immunosuppressive BALB/c mouse model. KEY FINDINGS: 98 monomer compounds in YPF were obtained. Through virtual analysis and screening on the oral utilization and drug likeness properties of the components, 47 effective components were got. 9 core targets obtained were enriched in IL-17 signaling pathway. In the mouse model, YPF could reduce the number of Can and alleviate Can-induced inflammation in the kidney effectively, upregulate Can-induced low proportion of CD4+/CD8+ of splenic lymphocytes, and increase Can-induced low activity of IL-17 pathway. SIGNIFICANCE: These results demonstrate that YPF could improve the immunity of Can-induced immunosuppression in BALB/c mice through upregulating the activity of IL-17 pathway.

Evaluation of In Vitro Immunomodulatory Activity of Withania somnifera Roots on Human Neutrophils.

The present study encompasses in vitro immunomodulatory activity of Withania somnifera WS (Solanaceae) examined on human neutrophils. The isolated compound of Withania somnifera roots was screened for its possible immunomodulatory activity by nitro blue tetrazolium test, phagocytosis of killed Candida albicans, neutrophil locomotion and chemotaxis. The extracted compound was examined at concentrations, viz. 10 µg/ml, 20 µg/ml, 40 µg/ml, 100 µg/ml and 1000 µg/ml. The activity expressed by the compound on human neutrophils was found to be significant for examined parameters.

Fungal sensitization and positive fungal culture from sputum in children with asthma are associated with reduced lung function and acute asthma attacks respectively.

BACKGROUND: Sensitization to thermotolerant fungi, including filamentous fungi and Candida albicans, is associated with poor lung function in adults with severe asthma. Data in children are lacking. Environmental exposure to fungi is linked with acute severe asthma attacks, but there are few studies reporting the presence of fungi in the airways during asthma attacks. METHODS: We investigated the association between fungal sensitization and/or positive fungal sputum culture and markers of asthma severity in children with chronic and acute asthma. Sensitization was determined using serum-specific IgE and skin prick testing against a panel of five fungi. Fungal culture was focused towards detection of filamentous fungi from sputum samples. RESULTS: We obtained sensitization data and/or sputum from 175 children: 99 with chronic asthma, 39 with acute asthma and 37 controls. 34.1% of children with chronic asthma were sensitized to thermotolerant fungi compared with no children without asthma (p =< 0.001). These children had worse pre-bronchodilator lung function compared with asthmatics without sensitization including a lower FEV1 /FVC ratio (p < .05). The isolation rate of filamentous fungi from sputum was higher in children with acute compared with chronic asthma. CONCLUSIONS: Fungal sensitization is a feature of children with chronic asthma. Children sensitized to thermotolerant fungi have worse lung function, require more courses of systemic corticosteroids and have greater limitation of activities due to asthma. Asthma attacks in children were associated with the presence of filamentous fungi positive sputum culture. Mechanistic studies are required to establish whether fungi contribute directly to the development of acute asthma.

Active neutrophil responses counteract Candida albicans burn wound infection of ex vivo human skin explants.

Burn wounds are highly susceptible sites for colonization and infection by bacteria and fungi. Large wound surface, impaired local immunity, and broad-spectrum antibiotic therapy support growth of opportunistic fungi such as Candida albicans, which may lead to invasive candidiasis. Currently, it remains unknown whether depressed host defenses or fungal virulence drive the progression of burn wound candidiasis. Here we established an ex vivo burn wound model, where wounds were inflicted by applying preheated soldering iron to human skin explants, resulting in highly reproducible deep second-degree burn wounds. Eschar removal by debridement allowed for deeper C. albicans penetration into the burned tissue associated with prominent filamentation. Active migration of resident tissue neutrophils towards the damaged tissue and release of pro-inflammatory cytokine IL-1ß accompanied the burn. The neutrophil recruitment was further increased upon supplementation of the model with fresh immune cells. Wound area and depth decreased over time, indicating healing of the damaged tissue. Importantly, prominent neutrophil presence at the infected site correlated to the limited penetration of C. albicans into the burned tissue. Altogether, we established a reproducible burn wound model of candidiasis using ex vivo human skin explants, where immune responses actively control the progression of infection and promote tissue healing.

Human Mesenchymal Stromal Cell Secretome Promotes the Immunoregulatory Phenotype and Phagocytosis Activity in Human Macrophages.

Human mesenchymal stromal/stem cells (hMSCs) show great promise in cell therapy due to their immunomodulatory properties. The overall immunomodulatory response of hMSCs resembles the resolution of inflammation, in which lipid mediators and regulatory macrophages (Mregs) play key roles. We investigated the effect of hMSC cell-cell contact and secretome on macrophages polarized and activated toward Mreg phenotype. Moreover, we studied the effect of supplemented polyunsaturated fatty acids (PUFAs): docosahexaenoic acid (DHA) and arachidonic acid, the precursors of lipid mediators, on hMSC immunomodulation. Our results show that unlike hMSC cell-cell contact, the hMSC secretome markedly increased the CD206 expression in both Mreg-polarized and Mreg-activated macrophages. Moreover, the secretome enhanced the expression of programmed death-ligand 1 on Mreg-polarized macrophages and Mer receptor tyrosine kinase on Mreg-activated macrophages. Remarkably, these changes were translated into improved Candida albicans phagocytosis activity of macrophages. Taken together, these results demonstrate that the hMSC secretome promotes the immunoregulatory and proresolving phenotype of Mregs. Intriguingly, DHA supplementation to hMSCs resulted in a more potentiated immunomodulation with increased CD163 expression and decreased gene expression of matrix metalloproteinase 2 in Mreg-polarized macrophages. These findings highlight the potential of PUFA supplementations as an easy and safe method to improve the hMSC therapeutic potential.

Mechanisms of microbial pathogenesis and the role of the skin microbiome in psoriasis: A review.

The pathogenesis of psoriasis may involve a breakdown of immune tolerance to cutaneous microorganisms. Psoriasis is associated with a higher incidence of Crohn disease and periodontitis, two diseases involving impaired tolerance and abnormal immune activation in response to intestinal and oral microbiota, respectively. In addition, guttate and chronic plaque psoriasis are associated with Streptococcus pyogenes colonization. The aim of this review is to characterize the microorganisms implicated in psoriasis by examining results of major association studies and possible mechanisms of pathogenesis. Although studies show relative increases in Streptococcus and Staphylococcus and decreases in Malassezia and Cutibacterium, they differ in methods of sampling and methods of microbial analysis. As such, no definitive associations between microbes and psoriasis have been found to date. It also remains unclear if changes in the microbiomal composition have a causal association with psoriasis or are simply a consequence of the inflammatory microenvironment. Techniques enabling strain-level analysis rather than species-level analysis of the skin microbiome are likely necessary to determine microbiomal signatures of psoriasis. Future investigations may lead to new diagnostic tests and novel treatments, such as probiotics or bacterial transplantation.

Efeitos do jato de plasma frio sob pressão atmosférica sobre Candida albicans / Cell effects of atmospheric pressure low temperature plasma jet on Candida albicans

As infecções causadas por Candida albicans são consideradas um desafio importante na área médica. Além dos antifúngicos convencionais apresentarem elevada toxicidade, casos de resistência de C. albicans a estes medicamentos têm sido descritos com frequência. Devido a estas limitações do uso de antifúngicos, a busca por terapias alternativas é alvo crescente de estudos. Dentre as novas terapias, o jato de plasma frio destaca-se a como agente eficaz frente a cepas de C. albicans, sendo considerada uma alternativa promissora. Seu efeito antifúngico é associado à presença de espécies reativas de oxigênio e nitrogênio liberadas, assim como de íons e fótons. Todavia até o presente momento, o exato mecanismo de ação não foi elucidado. Portanto, o objetivo deste estudo é prospectar os efeitos do jato de plasma frio sobre C. albicans. Perseguindo este objetivo, foram utilizadas duas técnicas complementares às técnicas microbiológicas clássicas, a espectroscopia no Infra-vermelho (FT-IR) e a microscopia de força atômica (AFM). Suspensões padronizadas da cepa C. albicans SC 5314 e 1 cepa clínica foram expostas ao jato de plasma. A seguir, foram analisadas por espectroscopia no infra-vermelho (FT-IR) e microscopia de força atômica (AFM) para identificar os efeitos do plasma sobre a parede celular de C. albicans. Controle negativo (sem exposição) e positivo (caspofungina) também foram analisados. Os ensaios foram realizados em triplicata em três experimentos independentes e a análise estatística foi realizada ao nível de significância de 5 %. As alterações morfológicas e topográficas nas células tratadas com plasma foram similares às do grupo tratado com caspofungina. O perfil bioquímico das células após tratamento com o plasma foi também similar ao das células tratadas com caspofungina. Conclui-se que o plasma agiu diretamente sobre as estruturas da parede celular fúngica, mais especificamente sobre os polissacarídeos, com provável influencia na redução de glucanos e aumento de mananas e quitinas. Esse remodelamento da parede celular pode ter correlação com as alterações morfológicas observadas, tais como mudança no tamanho da célula e aumento da rugosidade superficial(AU)

Infections caused by Candida albicans are considered important challenges in the medical area. Besides the high toxicity of conventional antifungals, cases of C. albicans resistance have been reported with increasing frequency. Due to these limitations of the antifungals, the search for alternative therapies is increasing. Among these therapies, cold plasma showed effectiveness against C. albicans isolates and is considered a promising alternative. Its antifungal effect is associated to the presence of oxygen and nitrogen reactive species, as well as ions. Nonetheless, the exact mechanism of action on C. albicans has not yet been elucidated. The aim of this study is to investigate the effects of plasma jet on C. albicans cell wall. For this purpose, two techniques complementing to classic the microbiological methods were used, the infrared spectroscopy (FT-IR) and atomic force microscopy (AFM). Standardized suspensions of C. albicans SC 5314 and 1 clinical isolate was exposed to plasma jet. Samples were analyzed by FT-IR and AFM to identifying the effects of plasma on C. albicans. Negative control (no exposition) and positive (caspofungin) were also analyzed. The experiments will be done in triplicate in three independent experiments and the statistical analyzes were done adopting the level of significance of 5%. The morphological and topographic alterations in cells treated with plasma were similar to the group treated with caspofungin. The biochemical profile of the cells after treatment with the plasma jet was also similar to those treated with caspofungin. It could be concluded that plasma acted directly on the structures of fungal cell wall, more specifically on polysaccharides, influencing the reduction of glucans and increased of mannans and chitins. This remodeling of cell wall can be correlated with the detected morphological alterations, such as changes in cell size and increase in superficial roughness(AU)

Influence of Acrylic Resin Denture Base Soaking Length in Siwak Extract Solution (Salvadora persica) on the Growth of Candida albicans

Objective: To know the influence of immersion length of denture base of acrylic resin in Siwak solution (Salvadora persica) on Candida albicans growth. Material and Methods: This type of research is laboratory experimental. The Siwak plant (Salvadora persica) was extracted and 1% solution was formed. The media used were Sabouraud Dextrose Broth (SDB) and Sabouraud Dextrose Agar (SDA). Candida albicans was cultured as 1 dose in 100 ml of SDB medium, and then incubated at shaker rotation for 1x24 hours. The concentration chosen to test the effectiveness of siwak extract solution was 1%, 10%, 15% and 25%. Data were presented as mean ± SD. Results: Zone of 6 hours Siwak extract immersion inhibitor by 43.47 ± 0.35, 8 hours to know the influence of immersion length of denture base of acrylic resin in Siwak solution (Salvadora persica) on candida albicans growth 44.42 ± 0.02, 10 hours of 52.79 ± 0.03. Conclusion: There is a difference of immersion length of denture base of acrylic resin on Candida albicans growth in Siwak extract solution (Salvadora persica).

Influência de extratos vegetais nos fatores de virulência de Candida albicans em biofilme: estudo in vitro / Plant extract and the virulence factors of Candida albicans

Cepas isoladas de infecções graves por Candida albicans expressam inúmeros fatores de virulência e a compreensão destes fatores permite esclarecer os mecanismos inerentes ao patógeno no processo infeccioso, assim como ferramentas terapêuticas que modulem a expressão destes fatores e permitam o controle de infecções persistentes. O objetivo deste estudo foi analisar a influência de 4 extratos vegetais no crescimento deste fungo quando em bifilme, assim como na expressão dos fatores de virulência (proteinase, fosfolipase e hemolisina) de C. albicans. Para isso, biofilmes de 48 horas de C. albicans (ATCC 18804) foram expostos a diferentes concentrações dos extratos glicólicos de Hamamelis virginiana, Persea. americana, Cynara scolymus L e Stryphnodendro barbatiman M, por 5 minutos e 24 h, para verificar a atividade antifúngica (redução em relação ao controle), e a secreção de proteinase, fosfolipase e hemolisina, utilizando meios especificos. Todos os extratos testados foram efetivos na redução do biofilme de C. albicans. Quando em contato por 5 min os extratos foram capazes de reduzir o biofilme em 50% em média, reduções mais expressivas foram encontradas depois de 24 h de exposição. O extrato de P. americana (25 mg/mL) reduziu o biofilme em 90%, em relação ao controle, seguido de C. scolymus (100 mg/mL), que reduziu em 85%. A secreção de proteinase foi alterada, tanto em 5 min como em 24 h, sendo a Pz (atividade enzimática) média de 0,69, em relação ao controle Pz= 0,49. Cynara scolymus foi o extrato que induziu a maior Pz média na concentração de 100 mg/mL, 0,73 e 0,78, quando tratados por 5 min e 24 h, respectivamente, em relação ao controle (Pz=0,48) (p<0,0001). A secreção de fosfolipase sofreu alteração depois do tratamento, sendo o extrato de S. barbatiman (100 mg/mL) o mais efetivo em 24 h, com Pz de 0,74 em relação ao controle (Pz= 0,50) (p<0,0001). A secreção de hemolisina foi alterada pela exposição ao extrato de H. virginiana (12,5 mg/mL) em 5 min de exposição. Em 24 h todos os extratos foram capazes de alterar o padrão hemolítico da cepa testada, sendo o extrato de H. virginiana (12,5 mg/mL) a apresentar Pz= 0,70. Os extratos vegetais testados demostram potencial antifúngico; e influenciam negativamente a secreção dos fatores de virulência testados, podendo ser apontados como ferramentas terapêuticas alternativas visando a redução da morbidade das infecções causadas por este patógeno (AU)

Strains isolated from severe Candida. albicans infections express numerous virulence factors, which suggest the understanding of these factors allows to clarify the mechanisms inherent to the pathogen in the infectious process, as well as therapeutic tools that modulate the expression of these factors and allow the control of persistent infections. Biofilms of 48 hours of C. albicans (ATCC 18804) were exposed to different concentrations of the glycolic extracts of Hamamelis virginiana and Persea. americana, Cynara. scolymus L and Stryphnodendron. barbatiman M, for 5 minutes and 24 h, to verify the antifungal activity, and secretion of proteinase, phospholipase and hemolysin. All extracts tested were effective in reducing the biofilm of C. albicans. When in contact for 5 min the extracts were able to reduce the mean of 50% of the biofilm. More significant reductions were found after 24 h of exposure. The extract of P. americana (25 mg/ml) reduced biofilm by 90% compared to control, followed by C. scolymus (100 mg/ml), which reduced by 85%. The enzymatic activiti (Pz) of proteinase was altered, both in 5 min and 24 h, the mean Pz being 0.69, in relation to the control Pz = 0.49. C. scolymus was the extract with the highest mean Pz at the concentration of 100 mg/ml, 0.73 and 0.78, when treated for 5 min and 24 h, respectively, in relation to the control (Pz = 0.48) (p <0,0001). Phospholipase secretion was altered after treatment, with S. barbatiman (100 mg/ml) being the most effective in 24 h, with Pz of 0.74 in relation to the control (Pz = 0.50) (p <0,0001). Hemolysin secretion was altered by exposure to H. virginiana (12.5 mg/ml) in 5 min of exposure. In 24 h all the extracts were able to change the hemolytic pattern of the tested strain, being the H. virginiana extract (12.5 mg/ml) to present Pz = 0,70. The tested plant extracts show antifungal potential; and negatively influence the secretion of the virulence factors tested, and can be indicated as alternative therapeutic tools aiming at reducing the morbidity of the infections caused by this pathogen(AU)

Fungal-derived cues promote ocular autoimmunity through a Dectin-2/Card9-mediated mechanism.

Uveitis (intraocular inflammation) is a leading cause of loss of vision. Although its aetiology is largely speculative, it is thought to arise from complex genetic-environmental interactions that break immune tolerance to generate eye-specific autoreactive T cells. Experimental autoimmune uveitis (EAU), induced by immunization with the ocular antigen, interphotoreceptor retinoid binding protein (IRBP), in combination with mycobacteria-containing complete Freund's adjuvant (CFA), has many clinical and histopathological features of human posterior uveitis. Studies in EAU have focused on defining pathogenic CD4+ T cell effector responses, such as those of T helper type 17 (Th17) cells, but the innate receptor pathways precipitating development of autoreactive, eye-specific T cells remain poorly defined. In this study, we found that fungal-derived antigens possess autoimmune uveitis-promoting function akin to CFA in conventional EAU. The capacity of commensal fungi such as Candida albicans or Saccharomyces cerevisae to promote IRBP-triggered EAU was mediated by Card9. Because Card9 is an essential signalling molecule of a subgroup of C-type lectin receptors (CLRs) important in host defence, we evaluated further the proximal Card9-activating CLRs. Using single receptor-deficient mice we identified Dectin-2, but not Mincle or Dectin-1, as a predominant mediator of fungal-promoted uveitis. Conversely, Dectin-2 activation by α-mannan reproduced the uveitic phenotype of EAU sufficiently, in a process mediated by the Card9-coupled signalling axis and interleukin (IL)-17 production. Taken together, this report relates the potential of the Dectin-2/Card9-coupled pathway in ocular autoimmunity. Not only does it contribute to understanding of how innate immune receptors orchestrate T cell-mediated autoimmunity, it also reveals a previously unappreciated ability of fungal-derived signals to promote autoimmunity.

Antifungal Activity, Phytochemical Characterization and Thermal Profile of Anadenanthera colubrina (Vell. ) Brenan

Objective: To investigate the antifungal potential of A. colubrina, and its phytochemical characteristics, thermal profile and toxicity. Material and Methods: To assess potential antifungal activity, the technique of microdilution was used with the determination of the Minimum Inhibitory Concentration and Minimum Fungicidal Concentration, using standard species of Candida and recent clinical isolates of Candida albicans. Analyses of action of the extract were performed on the wall and cell morphology of C. albicans, of the interactive effect between the plant extract and nystatin on C. albicans through the checkerboard method, and of growth kinetics. The phytochemical screening was determined by spectrophotometry. The thermal profile was traced with the determination of thermogravimetric curves (TG) and differential scanning calorimetry (DSC). The toxicity was evaluated by the method of hemolysis. Results: The extract of A. colubrina showed a fungistatic potential against all bacteria tested and it acted by modifying the cellular morphology of C. albicans. There was a synergism between nystatin and the plant extract (FIC=0.375), and 53.18% of total polyphenols were determined. The TG curve showed the occurrence of three steps of thermal decomposition. None of the tested concentrations became the effective cytotoxic concentration. Conclusion: Further studies should be conducted to understand the efficacy and the mechanisms of action involved in the antifungal activity of the plant extract of A. colubrina in order to produce a new drug for the treatment of oral candidiasis.

Immunochemical Cross-Reactivity of ß-Glucan in the Medicinal Plant, Sasa veitchii (Japanese Folk Medicine Kumazasa), and Medicinal Mushrooms.

Fungal ß-glucan is a representative pathogen-associated molecular pattern from mushroom, yeast, and fungi and stimulates innate as well as acquired immune systems. This ß-glucan is widely applied in functional food to enhance immunity. Humans and animals generally become sensitized to this ß-glucan and gradually produce specific antibodies to ß-glucans. The extracts of plants have been used as folk medicine and are reported to possess various biological activities that are beneficial for human health, such as antitumor, antiallergic, and anti-inflammatory activities. In the present study, the immunochemical cross-reactivity of Sasa extract and fungal ß-glucan was analyzed. We found that the anti-ß-glucan antibody in human sera strongly cross-reacted with the Sasa extract. This result strongly suggested that plant extracts modulate the immunostimulating effects of medicinal mushrooms. The cooperative effects of plants and mushrooms may be an important issue for functional foods.

Effectiveness of magnetic fluid hyperthermia against Candida albicans cells.

Candida albicans is one of the most frequently isolated fungal pathogens causing opportunistic infections in humans. Targeted magnetic fluid hyperthermia (MFH) is a promising method in thermal therapy facilitating selective heating of pathogen cells like C. albicans. In the paper, we used meso-2,3-dimercaptosuccinic acid (DMSA)-coated magnetic nanoparticles (MNPs) and functionalised anti-C. albicans immunomagnetic nanoparticles (IMNPs) to investigate the potential of MFH in combating C. albicans cells in vitro. Using Mössbauer spectroscopy it was found that synthesised MNPs exhibited superparamagnetic phenomena. On the basis of calorimetric experiments, the maximum SAR (specific absorption rate) was found and a proper concentration of MNPs was established to control the temperature. MFH based on both DMSA-coated MNPs and functionalised anti-C. albicans IMNPs was more effective in combating C. albicans cells in vitro than thermostat hyperthermia. Especially promising results were obtained using functionalised IMNPs, which eradicated most of the pathogen colonies at the temperature of 43 °C.

Activity of Extracts from Submerged Cultured Mycelium of Winter Mushroom, Flammulina velutipes (Agaricomycetes), on the Immune System In Vitro.

Extracts from submerged cultured mycelium of two strains of Flammulina velutipes, a popular culinary mushroom, were obtained by ultrasound and tested in vitro to determine their activity in innate immunity (monocytes/ macrophages). In addition, polyclonal antibodies against the extracts were produced. Both extracts have similar glycoproteins that contain mannose and glucose but have different glycoproteins with galactoseamine units. Two novel immunogenic glycoproteins with molecular weights of 32 and 25 kDa have been revealed. It is thought that these proteins are produced only by submerged cultured mycelium. Both extracts show immune-enhancing activity based on the significant modification of various parameters such as cytokine production, phagocytosis, and reactive oxygen species production.

Thymol has antifungal activity against Candida albicans during infection and maintains the innate immune response required for function of the p38 MAPK signaling pathway in Caenorhabditis elegans.

The Caenorhabditis elegans model can be used to study Candida albicans virulence and host immunity, as well as to identify plant-derived natural products to use against C. albicans. Thymol is a hydrophobic phenol compound from the aromatic plant thyme. In this study, the in vitro data demonstrated concentration-dependent thymol inhibition of both C. albicans growth and biofilm formation during different developmental phases. With the aid of the C. elegans system, we performed in vivo assays, and our results further showed the ability of thymol to increase C. elegans life span during infection, inhibit C. albicans colony formation in the C. elegans intestine, and increase the expression levels of host antimicrobial genes. Moreover, among the genes that encode the p38 MAPK signaling pathway, mutation of the pmk-1 or sek-1 gene decreased the beneficial effects of thymol's antifungal activity against C. albicans and thymol's maintenance of the innate immune response in nematodes. Western blot data showed the level of phosphorylation of pmk-1 was dramatically decreased against C. albicans. In nematodes, treatment with thymol recovered the dysregulation of pmk-1 and sek-1 gene expressions, the phosphorylation level of PMK-1 caused by C. albicans infection. Therefore, thymol may act, at least in part, through the function of the p38 MAPK signaling pathway to protect against C. albicans infection and maintain the host innate immune response to C. albicans. Our results indicate that the p38 MAPK signaling pathway plays a crucial role in regulating the beneficial effects observed after nematodes infected with C. albicans were treated with thymol.

Dysbiosis and Immune Dysregulation in Outer Space.

In space, the lifestyle, relative sterility of spaceship and extreme environmental stresses, such as microgravity and cosmic radiation, can compromise the balance between human body and human microbiome. An astronaut's body during spaceflight encounters increased risk for microbial infections and conditions because of immune dysregulation and altered microbiome, i.e. dysbiosis. This risk is further heightened by increase in virulence of pathogens in microgravity. Health status of astronauts might potentially benefit from maintaining a healthy microbiome by specifically managing their diet on space in addition to probiotic therapies. This review focuses on the current knowledge/understanding of how spaceflight affects human immunity and microbiome.

Regulation of mammalian siderophore 2,5-DHBA in the innate immune response to infection.

Competition for iron influences host-pathogen interactions. Pathogens secrete small iron-binding moieties, siderophores, to acquire host iron. In response, the host secretes siderophore-binding proteins, such as lipocalin 24p3, which limit siderophore-mediated iron import into bacteria. Mammals produce 2,5-dihydroxy benzoic acid, a compound that resembles a bacterial siderophore. Our data suggest that bacteria use both mammalian and bacterial siderophores. In support of this idea, supplementation with mammalian siderophore enhances bacterial growth in vitro. In addition, mice lacking the mammalian siderophore resist E. coli infection. Finally, we show that the host responds to infection by suppressing siderophore synthesis while up-regulating lipocalin 24p3 expression via TLR signaling. Thus, reciprocal regulation of 24p3 and mammalian siderophore is a protective mechanism limiting microbial access to iron.

Adjuvant effect of polysaccharide from fruits of Physalis alkekengi L. in DNA vaccine against systemic candidiasis.

Adjuvant effect mediated by polysaccharide (PPSB) isolated from the fruits of Physalis alkekengi L. in DNA vaccine was evaluated in mice. Recombinant plasmid containing epitope C (LKVIRK) from heat shock protein 90 (HSP90) of Candida albicans (C. albican) was used as DNA vaccine (pD-HSP90C). The results indicated that PPSB significantly enhanced specific antibody titers IgG, IgG1, IgG2b, and concentration of IL-2 and IL-4 in sera of mice immunized with pD-HSP90C (p<0.05). More importantly, it was found that the mice immunized with pD-HSP90C/PPSB not only had fewer CFU (colony forming unites) in the kidneys than mice immunized with pD-HSP90C, but also a statistically significant higher survival rate over PBS-injected group (p<0.05) when the immunized mice were challenged with living C. albican cells. However, no statistically significant difference in survival rate was observed between pD-HSP90C-immunized group and PBS-injected group. Therefore, PPSB can be considered as a promising adjuvant eliciting both Th1 and Th2 responses to enhance the efficacy of DNA vaccines.

Evaluation of Mdh1 protein as an antigenic candidate for a vaccine against candidiasis.

Candida albicans malate dehydrogenase (Mdh1p) has been screened by previous proteome studies as a candidate for a vaccine against candidiasis. In this study, recombinant Mdh1 protein with a His-tag was produced in Escherichia coli and evaluated as an immunogenic protein against candidiasis. Mdh1p was administrated to mice by two methods subcutaneous injection and intranasal administration before challenging them with a lethal dose of C. albicans. After vaccination of Mdh1p, antibody responses were observed. To evaluate the vaccination effect of Mdh1p, survival tests were performed after 35 d. Although all control mice died within 24 d or 25 d, 100% and 80% of mice survived with subcutaneous and intranasal administration, respectively. Therefore, our results indicate that, among C. albicans antigens examined thus far, Mdh1p is currently the most effective antigen for use as a vaccine for C. albicans.

Potencial antimicrobiano dos antissépticos de uso popular anapyon®, água rabelo® e malvatricin® sobre microrganismos do meio ambiente oral / Antimicrobial Potential of the Popular Antiseptics Anapyon®, Água Rabelo® and Malvatricin® against Oral Microorganisms

Objetivo: Verificar, ?in vitro?, o potencial antimicrobiano do Anapyon®, da Água Rabelo® e do Malvatricin® sobre microrganismos presentes na cavidade oral.Método: Para realização do experimento, utilizou-se o protocolo sequenciado durante quatro dias que avaliou, através da medida da densidade óptica, o potencial antimicrobiano dos fármacos nos microrganismos (Staphylococcus aureus, Candida tropicalis, Candida parapsilosis e Candida albicans), em suas formas planctônicas, apenas o Malvatricin® foi avaliado sobre as formas de biofilme por ser o único fármaco que apresentou resultados satisfatórios sobre as formas planctônicas. O estudo adotou como controle negativo a água destilada e controle positivo a Clorexidina®. Os resultados obtidos foram submetidos a uma análise estatística com os testes de Kruskal-Wallis e Mann-Whitney. Resultados: Observou-se, através da medida da densidade óptica, que apenas o Malvatricin® apresentou bons resultados na forma planctônica, sendo estes semelhantes ao controle positivo (clorexidina), considerado padrão nos ensaios antimicrobianos em Odontologia. Os resultados do Malvatricin® foram estatisticamente melhores quando comparados aos demais fármacos (Anapyon®, Água Rabelo®) e ao controle negativo. Este resultado foi semelhante para todos os microrganismos estudados: Staphylococcus aureus (p=0,002), Candida tropicalis (p=0,002), Candida parapsilosis (p=0,001) e Candida albicans (p<0,001). Desta forma, apenas o Mavatricin® foi testado para o microrganismo arranjado em biofilme. Observou-se, então, que para a C.albicans e para o S. aureus, houve diferença significativa entre clorexidina e Malvatricin® (p<0,05), com melhores resultados para o clorexidina. Para C. tropicalis, o Malvatricin® diferiu significativamente (p<0,05) da água destilada. Em relação a C.parapsilosis, nenhuma diferença foi observada em relação ao controle negativo (p=0,468). Conclusão: Apesar de alguns fármacos alternativos serem tidos como antimicrobianos, tais propriedades sobre células planctônicas e, principalmente, sobre biofilme foram observadas apenas para o Malvatricin®.

Objective: To evaluate in vitro the antimicrobial potential of Anapyon®, Água Rabelo and Malvatricin® against oral microorganisms.Method: The experiment used a four-day sequential protocol that evaluated by optical density measurements the antimicrobial potential of these products against Staphylococcus aureus, Candida tropicalis, Candida parapsilosis and Candida albicans in their planktonic forms. Only Malvatricin® was also evaluated against microbial biofilms because it was the only one to produce satisfactory results against the planktonic forms. Distilled water was used as negative control and Chlorhexidine® as positive control. The data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests. Results: According to the optical density readings, only Malvatricin® was effective against the planktonic microorganisms and its results were similar to those of the positive control (Chlorhexidine), which is the gold standard antimicrobial agent in dental research. Malvatricin® presented significantly higher efficacy than the other antiseptics (Anapyon® and Água Rabelo®) and the negative control, and this result was similar for all tested microorganisms: S. aureus (p=0.002), C tropicalis (p=0.002), C. parapsilosis (p=0.001) and C. albicans (p<0.001). For this reason, only Malvatricin® was evaluated against the microorganisms arranged as biofilms. Chlorhexidine presented significantly better results (p<0.05) than Malvatricin® against C. albicans and S. aureus. When tested against C. tropicalis, Malvatricin® differed significantly (p<0.05) from distilled water, while against C. parapsilosis no significant difference (p=0.468) was observed in comparison with the negative control. Conclusion: Although some pharmaceutical products being considered anti-microbial, such properties against planktonic cells and especially the biofilms were observed only for Malvatricin®.