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Eosinophils are mainly associated with parasitic infections and allergic manifestations. They produce many biologically active substances that contribute to the destruction of pathogens through the degranulation of microbicidal components and inflammatory tissue effects. In leishmaniasis, eosinophils have been found within inflammatory infiltrate with protective immunity against the parasite. We analyzed the responses of eosinophils from patients with localized (LCL) and diffuse (DCL) cutaneous leishmaniasis, as well as from healthy subjects, when exposed to Leishmania mexicana . All DCL patients exhibited blood eosinophilia, along with elevated eosinophil counts in non-ulcerated nodules. In contrast, only LCL patients with prolonged disease progression showed eosinophils in their blood and cutaneous ulcers. Eosinophils from DCL patients secreted significantly higher levels of IL-6, IL-8, and IL-13, compared to eosinophils from LCL patients. Additionally, DCL patients displayed higher serum levels of anti- Leishmania IgG antibodies. We also demonstrated that eosinophils from both LCL and DCL patients responded to L . mexicana promastigotes with a robust oxidative burst, which was equally intense in both patient groups and significantly higher than in healthy subjects. Coincubation of eosinophils (from donors with eosinophilia) with L . mexicana promastigotes in vitro revealed various mechanisms of parasite damage associated with different patterns of granule exocytosis: 1) localized degranulation on the parasite surface, 2) the release of cytoplasmic membrane-bound “degranulation sacs” containing granules, 3) release of eosinophil extracellular traps containing DNA and granules with major basic protein. In conclusion, eosinophils damage L . mexicana parasites through the release of granules via diverse mechanisms. However, despite DCL patients having abundant eosinophils in their blood and tissues, their apparent inability to provide protection may be linked to the release of cytokines and chemokines that promote a Th2 immune response and disease progression in these patients.

Background: Bacterial lysates are known to modulate the immune response against respiratory infections. However, the effects of the commercial bacterial lysate Pulmonarom® on dendritic cells—particularly human monocyte-derived dendritic cells (moDCs)—have not been studied. Additionally, limited data are available on the expression of Toll-like receptors (TLRs) and cytokines following stimulation with bacterial lysates. Methods: Human monocytes were isolated from buffy coats and differentiated into moDCs. Pulmonarom® was lyophilized, quantified, and used to stimulate moDCs. Ultrastructural changes were evaluated using transmission electron microscopy. The expression of TLRs and selected cytokines was analyzed by flow cytometry. Results: Pulmonarom® stimulation induced morphological changes in moDCs, including an increased number of dendrites and lysosomes. It also led to the upregulation of MHC class II molecules and TLRs 2, 3, 6, and 7. Additionally, the production of IL-4, IL-6, IL-8, and MCP-1 was significantly increased. Conclusions: Pulmonarom® promotes moDC maturation, characterized by enhanced antigen presentation capabilities and lysosomal activity, along with increased expression of specific TLRs and cytokines. These features suggest a trained immunity phenotype in moDCs, potentially improving their ability to initiate adaptive immune responses against respiratory pathogens. To our knowledge, this is the first study to investigate the immunomodulatory effects of Pulmonarom® on human moDCs, providing novel insights into its potential as an immunotherapeutic adjuvant.

Periodontal disease is considered one of the most prevalent chronic infectious diseases, often leading to the disruption of tooth-supporting tissues, including alveolar bone, causing tooth mobility and loss. is considered the major etiological agent of this disease, having a plethora of virulence factors, including, lipopolysaccharides (LPS), hemolysins, and proteinases. Antimicrobial peptides are one of the main components of the innate immune response that inhibit the growth of . The aim of this study was to analyze the antimicrobial activity of cystatin C and to assess the effect on the inflammatory and anti-inflammatory cytokines, the production of reactive oxygen species, and in the release of nitric oxide by human gingival fibroblasts incubated with in the presence and absence of cystatin C. ATCC 33277 was exposed to cystatin C for 24h and co-cultured with human gingival fibroblasts (HGFs) ATCC CRL-2014. The effect of cystatin on growth of and HGFs was evaluated. Pro-inflammatory (TNF , IL-1 ) and anti-inflammatory (IL-10) cytokines were determined by ELISA in the supernatants of HGFs incubated with exposed to cystatin C. Additionally, nitrites and reactive oxygen species (ROS) production were evaluated. Cystatin Cinhibited the growth of without affecting HGFs. Incubation of HGFs with led to a significant increase of TNF- and IL-1 . In contrast, HGFs incubated with exposed to cystatin C showed a decreased production of both cytokines, whereas IL-10 was enhanced. Incubation of HGFs with led to an increase of nitric oxide (NO) and ROS production, which was reduced in the presence of the peptide. Cystatin C inhibits the growth of P. gingivalis and decreases the inflammatory cytokines, ROS, and NO production during infection of HGFs with . Knowledge on the antimicrobial and immunomodulatory properties of cystatin C could aid in the design of new therapeutic approaches to facilitate the elimination of this bacterium to improve the treatment of periodontal disease.

Exosomes are extracellular microvesicles of endosomal origin (multivesicular bodies, MVBs) constitutively released by eukaryotic cells by fusion of MVBs to the plasma membrane. The exosomes from Leishmania parasites contain an array of parasite molecules such as virulence factors and survival messengers, capable of modulating the host immune response and thereby favoring the infection of the host. We here show that exosomes of L. mexicana amastigotes (aExo) contain the virulence proteins gp63 and PP2C. The incubation of aExo with bone marrow-derived macrophages (BMMs) infected with L. mexicana led to their internalization and were found to colocalize with the cellular tetraspanin CD63. Furthermore, aExo inhibited nitric oxide production of infected BMMs, permitting enhanced intracellular parasite survival. Expressions of antigen-presenting (major histocompatibility complex class I, MHC-I, and CD1d) and costimulatory (CD86 and PD-L1) molecules were modulated in a dose-dependent fashion. Whereas MHC-I, CD86 and PD-L1 expressions were diminished by exosomes, CD1d was enhanced. We conclude that aExo of L. mexicana are capable of decreasing microbicidal mechanisms of infected macrophages by inhibiting nitric oxide production, thereby enabling parasite survival. They also hamper the cellular immune response by diminishing MHC-I and CD86 on an important antigen-presenting cell, which potentially interferes with CD8 T cell activation. The enhanced CD1d expression in combination with reduction of PD-L1 on BMMs point to a potential shift of the activation route towards lipid presentations, yet the effectivity of this immune activation is not evident, since in the absence of costimulatory molecules, cellular anergy and tolerance would be expected.

Purpose Leishmania transmission by sand flies is detected by dermal cells that recognize ligands, such as lipophosphoglycan (LPG) on the promastigote glycocalyx. Resident dermal cells include γδ T cells, that recognize antigens by TCR or innate receptors, such as TLRs. We analyzed the response of dermal γδ T cells to Leishmania mexicana infections or inoculation of LPG, and whether parasite LPG activates γδ T cells through TLR2 . Methods We stimulated γδ T cells with LPG and analyzed colocalization of LPG and TLR2 by confocal microscopy. Activation of TLR2 was evaluated by IκBα phosphorylation. BALB/c mice were inoculated with L. mexicana or LPG in the dermis of earlobes, and LPG + TLR2 + γδ T cells were analyzed by flow cytometry. TNF + γδ T cells were examined in earlobe dermis by confocal microscopy. Results Stimulation with purified LPG showed activation of TLR2 with IκBα phosphorylation in γδ T cells. Inoculation of L. mexicana parasites or LPG into earlobe dermis showed co-expression of LPG + and TLR2 + in γδ T cells, demonstrating their interaction during infections. A subset of γδ T cells (LPG + and TLR2 − ) provided evidence that additional receptors recognize LPG. Inoculation of LPG enhanced overall γδ T cell numbers, including those expressing TNF, whereas infection with the parasite mostly enhanced γδ T cells expressing TNF. Conclusion L. mexicana LPG is a ligand recognized by TLR2 on γδ-T cells leading to their activation, although contribution of other receptors cannot be ruled out and need to be analyzed to elucidate their contribution during Leishmania infections.

Lantana camara (L.) is employed by several ethnical groups to treat numerous diseases. Although there are no ethnomedical reports on its use against leishmaniasis, organic extracts prepared from L. camara were shown to display leishmanicidal activity. In the present study, we carried out a bioassay-guided fractionation of the dichloromethane extract from Mexican L. camara in order to identify the compounds responsible for the leishmanicidal activity. Eighteen chromatographic fractions (FI–FXVIII) were evaluated in vitro against Leishmania mexicana and L. amazonensis. FII, FX, FXI, FXV, and FXVI showed significant activity against both Leishmania strains, the most potent of which was FXV. Eicosane (1), squalene (2), β-ionone (3), caryophyllene oxide (4), β-caryophyllene (5), hexanoic acid (6), tiglic acid (7), a mixture of lantanilic (8) and camaric (9) acids, and lantadene B (10) were identified and obtained from the active fractions and evaluated for their leishmanicidal activity. The mixture of lantanilic (8) and camaric (9) acids (79%/21%) was the most potent one (half maximal inhibitory concentration (IC50) = 12.02 ± 0.36 μM). This study indicates that this cultivar of L. camara has high potential for the development of phytomedicines or as a source of natural products, which might represent lead compounds for the design of new drugs against leishmaniasis.

The propolis produced by stingless bees is a complex mixture of natural sticky components mixed with soil or clay. Global research on propolis has focused on studying the biological and pharmacological properties and chemical composition of stingless bee propolis from Brazil, Indonesia, and other regions. However, studies of stingless bee propolis produced in Mexico are scarce. This study aimed to determine the chemical composition of the geopropolis of Scaptotrigona mexicana collected in the Totonacapan region and to evaluate its antioxidant and antibacterial activities. The phenolic contents of the ethanolic extract of the collected propolis ranged from 2.45 ± 0.03 mg GAE/g to 3.48 ± 0.56 mg GAE/g of dry extract. The total flavonoid content ranged from 0.69 ± 0.03 mg QE/g to 0.84 ± 0.009 mg QE/g of dry extract. The antioxidant activity of the ethanolic extracts was assessed via DPPH, ABTS, and FRAP assays. The minimum inhibitory concentration values exhibited by the ethanolic extract (>512 g/mL) for Gram-negative bacteria (Pseudomonas aerugunosa and Phorphyromonas gingivalis) were higher than those of Gram-positive bacteria. The stingless bee propolis extract showed the highest antibacterial activity against Streptococcus mutans (256 g/mL). Five known compounds, taraxeryl acetate (1), lupeol (3), cicloart-23-en-3β,25-diol (5), mangiferoic acid (6), and 5-(11’Z-heptadecenyl)-resorcinol (7), and two irresoluble mixtures of 3-O-acetyl-α-(2a) and 3-O-acetyl-β-amyrins (2b), and α- (4a) and -amyrins (4b), were identified by nuclear magnetic resonance spectroscopy and mass spectrometry. Additionally, 39 volatile compounds were identified via headspace-solid phase microextraction using the hyphenated gas chromatography coupled to mass spectrometry time-of-flight. The main volatile compounds detected include trans-α-bergamotene (8.15%), hexanal (7.17%), 2-heptanone (7.60%), and α-copaene (7.09%).

Eosinophils are mainly associated with parasitic infections and allergic manifestations. They produce many biologically active substances that contribute to the destruction of pathogens through the degranulation of microbicidal components and inflammatory tissue effects. In leishmaniasis, eosinophils have been found within inflammatory infiltrate with protective immunity against the parasite. We analyzed the responses of eosinophils from patients with localized (LCL) and diffuse (DCL) cutaneous leishmaniasis, as well as from healthy subjects, when exposed to Leishmania mexicana. All DCL patients exhibited blood eosinophilia, along with elevated eosinophil counts in non-ulcerated nodules. In contrast, only LCL patients with prolonged disease progression showed eosinophils in their blood and cutaneous ulcers. Eosinophils from DCL patients secreted significantly higher levels of IL-6, IL-8, and IL-13, compared to eosinophils from LCL patients. Additionally, DCL patients displayed higher serum levels of anti-Leishmania IgG antibodies. We also demonstrated that eosinophils from both LCL and DCL patients responded to L. mexicana promastigotes with a robust oxidative burst, which was equally intense in both patient groups and significantly higher than in healthy subjects. Coincubation of eosinophils (from donors with eosinophilia) with L. mexicana promastigotes in vitro revealed various mechanisms of parasite damage associated with different patterns of granule exocytosis: 1) localized degranulation on the parasite surface, 2) the release of cytoplasmic membrane-bound \"degranulation sacs\" containing granules, 3) release of eosinophil extracellular traps containing DNA and granules with major basic protein. In conclusion, eosinophils damage L. mexicana parasites through the release of granules via diverse mechanisms. However, despite DCL patients having abundant eosinophils in their blood and tissues, their apparent inability to provide protection may be linked to the release of cytokines and chemokines that promote a Th2 immune response and disease progression in these patients.

This proposal evaluates the energy potential of agricultural residues of Zea mays from an indigenous community in Mexico. The study consists of four stages: (a) evaluation of residue production in all community farming areas (b) morphological and physicochemical characterization, using scanning electron microscopy (SEM), as well as infrared spectroscopy (FTIR) and Raman (c) the proximal and functional evaluation of the residues, through fiber analysis, determination of fixed carbon, humidity, estimation of calorific value, ash microanalysis and elemental analysis, and (d) evaluation of energy potential and multicriteria analysis. The results show that Z. mays residues have initial moisture values of less than 10%, ash content below 20%, fixed carbon around 14% and a calorific value of 17.6 MJ/kg associated with polymeric compounds and carbohydrates, as well as a percentage of extractable compounds of the order of 40%. The production of these residues on the 249 hectares (ha) of cultivation used would generate 23 TJ/year, whereas if the total number of hectares available were cultivated, the total energy generation would be 330 TJ/year, which is enough to satisfy the wood fuel demand of approximately seven communities with the characteristics of the study community. Due to this potential, as well as the results of the characterization, the agricultural mentioned residues are an energy alternative to meet the energy demand in communities in Michoacán, Mexico.

Anthelmintic resistance is currently negatively impacting animal production parameters, leading to an increase in the prevalence of gastrointestinal nematodes and resulting in low profitability in small ruminants. Therefore, there is a need to develop alternative control strategies to reduce the prevalence and damage caused by these parasites in extensive systems. One of these strategies involves plant extracts and their secondary metabolites, which have shown antiparasitic properties. The main aim of the present study was the evaluation of Artemisia cina (A. cina) foliage to perform an n-hexane extract and cinaguaiacin as secondary metabolite (mixture of 63% of 3′-demethoxy-6-O-demethylisoguaiacin and 37% norisoguaiacin), previously identified by chromatography technique and relative expression of three antioxidant enzyme genes of infective Haemonchus contortus larvae (L3). The results showed upregulation of glutathione peroxidase (GPx) and catalase (CAT), and decreased expression of superoxide dismutase (SOD) genes after exposure to H. contortus L3 to n-hexane extract of A. cina. Furthermore, cinaguacin displayed up- and downregulation of GPx and superoxide dismutase genes, respectively. These data suggest the active function of reactive oxidative species (ROS) genes of H. contortus L3 exposed by the extract of A. cina and cinaguaiacin to induce the larve death. In this sense, both alternatives could be promising to mitigate resistance to anthelmintic drugs.