ABSTRACT
Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype. Currently, paclitaxel (PTX) represents the first-line therapy for TNBC; however it presents a hydrophobic behavior and produces severe adverse effects. The aim of this work is to improve the therapeutic index of PTX through the design and characterization of novel nanomicellar polymeric formulations composed of a biocompatible copolymer Soluplus® (S), surface-decorated with glucose (GS), and co-loaded either with histamine (HA, 5 mg/mL) and/or PTX (4 mg/mL). Their micellar size, evaluated by dynamic light scattering, showed a hydrodynamic diameter between 70 and 90 nm for loaded nanoformulations with a unimodal size distribution. Cytotoxicity and apoptosis assays were performed to assess their efficacy in vitro in human MDA-MB-231 and murine 4T1 TNBC cells rendering optimal antitumor efficacy in both cell lines for the nanoformulations with both drugs. In a model of TNBC developed in BALB/c mice with 4T1 cells, we found that all loaded micellar systems reduced tumor volume and that both HA and HA-PTX-loaded SG micelles reduced tumor weight and neovascularization compared with the empty micelles. We conclude that HA-PTX co-loaded micelles in addition to HA-loaded formulations present promising potential as nano-drug delivery systems for cancer chemotherapy.
Subject(s)
Antineoplastic Agents, Phytogenic , Triple Negative Breast Neoplasms , Mice , Humans , Animals , Paclitaxel , Histamine , Micelles , Antineoplastic Agents, Phytogenic/chemistry , Cell Line, Tumor , Polyethylene Glycols/chemistry , Polymers , Drug Carriers/chemistry , Mice, Inbred BALB CABSTRACT
BACKGROUND: Shigella specie is a globally important intestinal pathogen disseminated all over the world. In this study we analyzed the genome and the proteomic component of two Shigella flexneri 2a clinical isolates, collected from pediatric patients with gastroenteritis of the Northwest region of Argentina (NWA) in two periods of time, with four years of difference. Our goal was to determine putative changes at molecular levels occurred during these four years, that could explain the presence of this Shigella`s serovar as the prevalent pathogen in the population under study. RESULTS: As previously reported, our findings support the idea of Shigella has a conserved "core" genome, since comparative studies of CI133 and CI172 genomes performed against 80 genomes obtained from the NCBI database, showed that there is a large number of genes shared among all of them. However, we observed that CI133 and CI172 harbors a small number of strain-specific genes, several of them present in mobile genetic elements, supporting the hypothesis that these isolates were established in the population by horizontal acquisition of genes. These differences were also observed at proteomic level, where it was possible to detect the presence of certain secreted proteins in a culture medium that simulates the host environment. CONCLUSION: Great similarities were observed between the CI133 and CI172 strains, confirming the high percentage of genes constituting the "core" genome of S. flexneri 2. However, numerous strain specific genes were also determined. The presence of the here identified molecular elements into other strain of our culture collation, is currently used to develop characteristic markers of local pathogens. In addition, the most outstanding result of this study was the first description of a S. flexneri 2 producing Colicin E, as one of the characteristics that allows S. flexneri 2 to persist in the microbial community. These findings could also contribute to clarify the mechanism and the evolution strategy used by this pathogen to specifically colonize, survive, and cause infection within the NWA population.
Subject(s)
Dysentery, Bacillary , Shigella , Argentina/epidemiology , Child , Genomics , Humans , Infant , Proteomics , Shigella flexneri/geneticsABSTRACT
The discovery of the human histamine H4 receptor (H4R) has contributed to our understanding of the role of histamine in numerous physiological and pathological conditions, including tumor development and progression. The lymph nodes of patients with malignant lymphomas have shown to contain high levels of histamine, however, less is known regarding the expression and function of the H4R in T-cell lymphoma (TCL). In this work we demonstrate the expression of H4R isoforms (mRNA and protein) in three human aggressive TCL (OCI-Ly12, Karpas 299, and HuT78). Histamine and specific H4R agonists (VUF8430 and JNJ28610244) significantly reduced cell viability in a dose-dependent manner (p < 0.05). The combined treatment with the H4R antagonist (JNJ7777120, 10 µM) reversed the effects of the H4R ligands. Importantly, we screened a drug repurposing library of 433 FDA-approved compounds (1 µM) in combination with histamine (10 µM) in Hut78 cells. Histamine produced a favorable antitumor effect with 18 of these compounds, including the histone deacetylase inhibitor panobinostat. Apoptosis, proliferation, and oxidative stress studies confirmed the antitumoral effects of the combination. We conclude that the H4R is expressed in TCL, and it is involved in histamine-mediated responses.
Subject(s)
Antineoplastic Agents/pharmacology , Histamine Agonists/pharmacology , Lymphoma, T-Cell/drug therapy , Receptors, Histamine H4/metabolism , Apoptosis/drug effects , Cell Line , Cell Line, Tumor , Cell Proliferation/drug effects , HEK293 Cells , Histamine/metabolism , Histamine Antagonists/pharmacology , Humans , Lymphoma, T-Cell/metabolism , Oxidative Stress/drug effectsABSTRACT
Resumen Introducción. El tumor fibroso solitario de pleura (TFSP) es una neoplasia poco frecuente, con aproximadamente 1.000 casos reportados en la literatura mundial. La aproximación diagnóstica inicial se realiza con estudios imagenológicos. Métodos. De forma retrospectiva, se recopilaron cuatro casos de pacientes con TFSP gigante operados en nuestra institución. Se describen las características sociodemográficas, clínicas, imagenológicas, macroscópicas y microscópicas de cada caso. Resultados. Todos los pacientes de la serie cursaron con manifestaciones clínicas, con un promedio de 23,75 meses de evolución. El 50% de los tumores se localizaron en la cavidad pleural derecha y el 50% en la izquierda. En tomografía computarizada (TC) de tórax, los cuatro casos se presentaron como una masa sólida, de densidad heterogénea, con diámetros mayores entre 17 y 22 cm y contornos variables (lisos en tres casos y lobulados en un paciente). Se observaron calcificaciones intratumorales en dos casos y derrame pleural en tres pacientes. En cirugía, todas las masas presentaron pedículos. El análisis histológico e inmuno-químico confirmó la naturaleza benigna de tres casos y malignidad en una de las neoplasias. Conclusiones. Los TFSP generalmente son benignos y de buen pronóstico. Sin embargo, entre 10 y 20% de esos tumores son malignos. Las imágenes diagnósticas pueden sugerir el diagnóstico de TFSP, pero la confirmación de la naturaleza de la lesión debe realizarse con el análisis histopatológico de toda la pieza quirúrgica.
Abstract Introduction. Solitary fibrous tumor of the pleura (SFTP) is a rare neoplasm, with ~1.000 cases reported in the worldwide literature. The initial diagnostic approach is performed by imaging studies. Methods. Retrospectively, we collected four cases of patients with giant SFTP operated in our institution. The sociodemographic, clinical, imaging, macroscopic, and microscopic characteristics of each case are described. Results. All the patients in the series had clinical manifestations, with an average of 23.75 months of evolution. 50% of the tumors were located in the right pleural cavity and 50% in the left. In chest computed tomography (CT), the four cases presented as a solid mass, of heterogeneous density, with greater diameters between 17 and 22 cm, and variable contours (smooth in three cases and lobulated in one patient). Intratumoral calcifications were observed in two cases and pleural effusion in three patients. In surgery, all masses presented pedicles. The histological and immunochemical analysis confirmed the benign nature of three cases and malignancy in one of them. Conclusions. SFTPs are usually benign and have a good prognosis. However, between 10 and 20% of these tumors are malignant. Diagnostic images may suggest the diagnosis of SFTP, but confirmation of the nature of the lesion should be made with the histopathological analysis of the entire surgical specimen.
ABSTRACT
Cancer is the second leading cause of death globally and its incidence and mortality are rapidly increasing worldwide. The dynamic interaction of immune cells and tumor cells determines the clinical outcome of cancer. Immunotherapy comes to the forefront of cancer treatments, resulting in impressive and durable responses but only in a fraction of patients. Thus, understanding the characteristics and profiles of immune cells in the tumor microenvironment (TME) is a necessary step to move forward in the design of new immunomodulatory strategies that can boost the immune system to fight cancer. Histamine produces a complex and fine-tuned regulation of the phenotype and functions of the different immune cells, participating in multiple regulatory responses of the innate and adaptive immunity. Considering the important actions of histamine-producing immune cells in the TME, in this review we first address the most important immunomodulatory roles of histamine and histamine receptors in the context of cancer development and progression. In addition, this review highlights the current progress and foundational developments in the field of cancer immunotherapy in combination with histamine and pharmacological compounds targeting histamine receptors.
Subject(s)
Histamine/metabolism , Neoplasms/metabolism , Receptors, Histamine/metabolism , Tumor Microenvironment/immunology , Adaptive Immunity/immunology , Antineoplastic Agents, Immunological/therapeutic use , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Basophils/immunology , Basophils/metabolism , Histamine/immunology , Humans , Immunity, Innate/immunology , Immunotherapy , Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Macrophages/immunology , Macrophages/metabolism , Mast Cells/immunology , Mast Cells/metabolism , Neoplasms/immunology , Neoplasms/therapy , Neutrophils/immunology , Neutrophils/metabolism , Receptors, Histamine/immunology , T-Lymphocytes/immunology , T-Lymphocytes/metabolismABSTRACT
This paper presents data on free-living ticks collected by flagging and using CO2 traps in three natural areas in Costa Rica: Carara National Park (CNP), Palo Verde National Park (PVNP), and a Private Forest Reserve in Sarapiquí (SPR). Data were analyzed calculating aspects of alpha diversity (species richness, entropy; dominance index, and evenness); and for beta diversity, compositional similarity between communities of ticks was also calculated. We collected 12,795 ticks belonging to 10 species: Amblyomma coelebs, Amblyomma dissimile, Amblyomma mixtum, Amblyomma naponense, Amblyomma cf. oblongoguttatum, Amblyomma cf. parvum, Amblyomma sabanerae, Amblyomma tapirellum, Haemaphysalis juxtakochi and Ixodes affinis. The number of species and individuals varied between sites: 5970 ticks were collected in CNP, 4443 in PVNP, and 2382 in SPR. Amblyomma cf. oblongoguttatum and A. cf. parvum were collected at all three sites, but A. mixtum was the most abundant species, even though it was not collected in SPR. Values of alpha diversity were calculated for CNP and SPR, while diversity in PVNP was the lowest of the three locations. Evenness was highest in SPR and lowest in CNP. The only community that presented dominance was PVNP. Beta diversity showed low similarity between the three locations with the lowest being CNP and SPR. For the three localities, estimates of the number of tick species based on presence/absence data was higher using flagging than CO2; and considering the stage of the ticks collected. More larvae were captured using CO2 traps than by flagging, while flagging was better for collecting adults. To our knowledge this is the first study in Costa Rica that compares these two sampling methods in three different environmental areas.
Subject(s)
Animal Distribution , Biodiversity , Ixodes/physiology , Animals , Costa Rica , Environment , Ixodes/growth & development , Larva/growth & development , Larva/physiology , Nymph/growth & development , Nymph/physiology , Parks, RecreationalABSTRACT
BACKGROUND: The histamine H4 receptor (H4R) is preferentially expressed in immune cells and is a potential therapeutic target for inflammatory and autoimmune diseases. This study aimed at further exploring the role of H4R in the immunobiology of breast cancer. METHODS: We used wild type (WT) and H4R deficient mice (KO) to evaluate whether H4R genotypes show a different distribution of T cell subsets in spleens, tumours and tumour draining lymph nodes (TDLN) in a syngeneic ErbB2-positive breast cancer model developed orthotopically with LM3 cells and its impact on tumour growth. RESULTS: The presence of tumours had a differential impact on the distribution of T cells in TDLN from KO mice compared to WT ones. At day 21 post-inoculation (p.i.) of cells, despite no significant changes in the tumour weight, TDLN from KO mice showed a significantly increased proportion of CD8+ T cells compared to WT mice. At day 38 p.i. of cells a reduced tumour weight was evident in KO mice. This was accompanied by a decreased proportion of CD4+CD25+FoxP3+ regulatory T cells in TDLN of KO compared to WT mice. Tumour-bearing KO mice showed a better survival compared to WT mice. CONCLUSIONS: H4R-mediated mechanisms may modulate the immune tumour microenvironment, promoting an immunosuppressive milieu. Results suggest that H4R could be explored as an immunotherapeutic target with potential benefit in combination with immunotherapy. Further preclinical and clinical studies are necessary to confirm this hypothesis.
Subject(s)
Breast Neoplasms/immunology , Breast Neoplasms/metabolism , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Receptors, Histamine H4/deficiency , Receptors, Histamine H4/immunology , T-Lymphocytes, Regulatory/immunology , Animals , CD4-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/metabolism , Disease Models, Animal , Female , Histamine/immunology , Interleukin-2 Receptor alpha Subunit/immunology , Mice , Mice, Inbred BALB C , Mice, Knockout , T-Lymphocytes, Regulatory/metabolism , Tumor Microenvironment/immunologyABSTRACT
Northwest Argentina (NWA) is a poor economic-geographical region, with the highest rate of diarrhea diseases. At the moment, there are no reports showing the epidemiological status of this region that would allow to establish methods for prevention and control of these infections and to indicate of the prevalent pathogen that produces them. Therefore we carried out an epidemiological study of the gastroenteritis etiological agents and their incidence in the pediatric population. A total of 17 823 fecal samples were collected, 14 242 from HNJ-Tuc, 2,257 from CePSI-Stgo and 1,324 from HINEP-Cat. In 2,595 samples a bacterial agent was identified, the 93.64% corresponded to Shigella/Salmonella clinical isolates. Shigella genus was the prevalent pathogen, being Shigella flexneri 2 the most frequent serotype. Most of the Shigella clinical isolates presented themselves as multidrug-resistant (MDR), harboring 2 to 3 genetic resistance determinants. 50% of the affected patients were children under 4 years old. Here, we demonstrate that bacterial gastrointestinal diseases strongly affect the health of NWA population. The appearance of epidemic outbreaks, as happened during 2014, suggest that they may be related to the socio-economic poverty of NWA. Recently, Shigella flexneri 2 has become the highest NWA´s incidence infectious agent. The acquisition of new antibiotic resistance determinants may play an important role in their adaptation and persistence.
Subject(s)
Bacterial Infections/epidemiology , Diarrhea/microbiology , Salmonella/isolation & purification , Shigella/isolation & purification , Adolescent , Argentina/epidemiology , Child , Child, Preschool , Diarrhea/epidemiology , Drug Resistance, Multiple, Bacterial , Dysentery, Bacillary/epidemiology , Epidemiologic Studies , Feces/microbiology , Female , Humans , Infant , Infant, Newborn , Male , Multiplex Polymerase Chain Reaction , Poverty , Prevalence , Salmonella/classification , Salmonella/genetics , Salmonella Infections/epidemiology , Serogroup , Shigella/classification , Shigella/geneticsABSTRACT
Epithelial-mesenchymal transition (EMT) contributes to cell invasion and metastasis during the progression of epithelial cancers. Though preclinical evidence suggests a role for histamine H4 receptor (H4R) in breast cancer growth, its function in the EMT is less known. In this study we proposed to investigate the effects of H4R ligands on EMT and mammosphere formation as a surrogate assay for cancer stem cells in breast cancer cells with different invasive phenotype. We also investigated the participation of Src and TGF-ß signaling in these events. Breast cancer cells were treated with the H4R agonists Clobenpropit, VUF8430 and JNJ28610244 and the H4R antagonist JNJ7777120. Immunodetection studies showed cytoplasmic E-cadherin, cytoplasmic and nuclear beta-catenin, nuclear Slug and an increase in vimentin and α-smooth muscle actin expression. There was also an enhancement in cell migration and invasion assessed by transwell units. All these effects were prevented by JNJ7777120. Moreover, H4R agonists induced an increase in phospho-Src levels detected by Western blot. Results revealed the involvement of phospho-Src in EMT events. Upon treatment with H4R agonists there was an increase in phospho-ERK1/2 and TGF-ß1 levels by Western blot, in Smad2/3 positive nuclei by indirect immunofluorescence, and in tumor spheres formation by the mammosphere assay. Notably, the selective TGF-ß1 kinase/activin receptor-like kinase inhibitor A83-01 blocked these effects. Moreover, cells derived from mammospheres exhibited higher Slug expression and enhanced migratory behavior. Collectively, findings support the interaction between H4R and TGF-ß receptor signaling in the enhancement of EMT features and mammosphere formation and point out intracellular TGF-ß1 as a potential mediator of these events.
Subject(s)
Breast Neoplasms/metabolism , Epithelial-Mesenchymal Transition/physiology , Oncogene Protein pp60(v-src)/metabolism , Receptors, Histamine H4/agonists , Receptors, Histamine H4/metabolism , Transforming Growth Factor beta1/metabolism , Epithelial-Mesenchymal Transition/drug effects , Female , Humans , Indoles/pharmacology , MCF-7 Cells , Piperazines/pharmacologyABSTRACT
BACKGROUND: The aim of this work was to improve the knowledge of the role of histamine in breast cancer by assessing the therapeutic efficacy of histamine and histamine H4 receptor (H4R) ligands in a triple-negative breast cancer (TNBC) model developed in immunocompetent hosts. By using publicly available genomic data, we further investigated whether histidine decarboxylase (HDC) could be a potential biomarker. METHODS: Tumours of 4T1 TNBC cells were orthotopically established in BALB/c mice. Treatments employed (mg kg-1): histamine (1 and 5), JNJ28610244 (H4R agonist, 1 and 5) and JNJ7777120 (H4R antagonist, 10). RESULTS: Increased HDC gene expression is associated with better relapse-free and overall survival in breast cancer patients. Histamine treatment (5 mg kg-1) of 4T1 tumour-bearing mice reduced tumour growth and increased apoptosis. Although no immunomodulatory effects were observed in wild-type mice, significant correlations between tumour weight and cytotoxic lymphocyte infiltration were detected in H4R knockout mice. H4R agonist or antagonist differentially modulated tumour growth and immunity in 4T1 tumour-bearing mice. CONCLUSIONS: Histamine plays a complex role and stands out as a promising drug for TNBC treatment, which deserves to be tested in clinical settings. HDC expression level is associated with clinicopathological characteristics, suggesting a prognostic value in breast cancer.
Subject(s)
Apoptosis/drug effects , Cell Proliferation/drug effects , Histamine Agonists/pharmacology , Histamine Antagonists/pharmacology , Histidine Decarboxylase/metabolism , Receptors, Histamine H4/genetics , Receptors, Histamine H4/metabolism , Triple Negative Breast Neoplasms/metabolism , Animals , Breast Neoplasms/metabolism , Breast Neoplasms/mortality , Cell Line, Tumor , Cell Survival/drug effects , Databases, Factual , Female , Histamine/pharmacology , Humans , Indoles/pharmacology , Lymphocytes, Tumor-Infiltrating/drug effects , Mice , Mice, Inbred BALB C , Mice, Knockout , Oximes/pharmacology , Piperazines/pharmacology , Prognosis , Triple Negative Breast Neoplasms/mortality , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
The virulence genes of Salmonella are modulated during infection by several regulatory systems, and the RcsCDB system is one of the most important of these. The S. Typhimurium EG14873 (rcsC11) strain harbours the rcsC11 point mutation, displaying a constitutive activation of this system, which is characterized by mucoid colonies and attenuated virulence phenotypes. In this work, the stability of the rcsC11 mutation was analysed under stress conditions. Under acid and anaerobic stresses, we observed the appearance of small and non-mucoid colonies of the rcsC11 strain. The sequencing of the rcsC gene from these colonies showed that the mutation is conserved. Moreover, we found that small colonies were also generated when the wild-type strain grew in acid and anaerobic conditions. It is worth noting that the transition from normal to atypical colonies of both strains only took place after several days of incubation and was not observed during eukaryotic cell infection. Therefore, the appearance of these atypical colonies is a characteristic feature of S. Typhimurium strains under stressful situations and does not involve a reversion of the rcsC11 allele and nor does it imply any risk to mammalian cells. Therefore, we propose that the S. Typhimurium rcsC11 strain is a good candidate for the development of attenuated vaccines.
Subject(s)
Bacterial Proteins/genetics , Mutation , Salmonella typhimurium/physiology , Stress, Physiological , Acids/metabolism , Anaerobiosis , Animals , Mice , Phenotype , RAW 264.7 Cells , Salmonella typhimurium/genetics , Salmonella typhimurium/growth & development , Salmonella typhimurium/pathogenicity , Vaccines, Attenuated/genetics , Virulence/geneticsABSTRACT
In Salmonella enterica serovar Typhimurium, the RcsCDB regulatory system controls the expression of genes involved in synthesis of colanic acid, formation of flagella and virulence. Here, we show that activation of the RcsCDB system downregulates expression of std, an operon that encodes fimbriae involved in Salmonella attachment to the mucus layer in the large intestine. Bioinformatic analysis predicts the existence of an RcsB-binding site located 180 bp upstream to the +1 transcription start site of the std promoter, and electrophoretic mobility shift assays confirm that RcsB binds the std promoter region in vitro. This study adds RcsB to the list of regulators of std transcription and provides an example of modulation of fimbriae synthesis by a signal transduction system.
Subject(s)
Bacterial Proteins/metabolism , Fimbriae Proteins/genetics , Fimbriae, Bacterial/genetics , Gene Expression Regulation, Bacterial , Salmonella typhimurium/metabolism , Signal Transduction , Bacterial Adhesion , Bacterial Proteins/genetics , Binding Sites , Mutation , Operon , Promoter Regions, Genetic , Salmonella typhimurium/genetics , Transcription, GeneticABSTRACT
The intracellular pathogen Salmonella is an important cause of human foodborne diseases worldwide. Salmonella takes advantage of the phosphorelay regulatory systems to survive in the hostile environment of the host's gastrointestinal tract. It has been reported that the nitrate reductase Z (NR-Z), encoded by the narUZYV operon, is required during Salmonella transition to anaerobic environments and is constitutively produced at low levels, but little is known about the regulatory mechanism involved in the operon gene expression. In this work, we found that the RcsCDB system is activated by high concentrations of specific sugars as a carbon source. In this activation state, the RcsCDB system participates in the negative control of narUZYWV expression. This control strategy occurs during exponential growth when the carbon source is high, allowing for normal aerobic respiration. The RcsCDB system's participation in aerobic respiration is necessary to ensure efficient metabolism and optimal energy consumption when the bacteria are growing exponentially.
Subject(s)
Bacterial Proteins/genetics , Nitrate Reductase/genetics , Salmonella typhimurium/growth & development , Salmonella typhimurium/genetics , Transcription Factors/physiology , Gene Expression Regulation, Bacterial , Operon , Promoter Regions, Genetic , Transcription, GeneticABSTRACT
Bacterial survive and respond to adverse changes in the environment by regulating gene transcription through two-component regulatory systems. In Salmonella Typhimurium the STM1485 gene expression is induced under low pH (4.5) during replication inside the epithelial host cell, but it is not involved in sensing or resisting to this condition. Since the RcsCDB system is activated under acidic conditions, we investigated whether this system is able to modulate STM1485 expression. We demonstrated that acid-induced activation of the RcsB represses STM1485 transcription by directly binding to the promoter. Under the same condition, the RstA regulator activates the expression of this gene. Physiologically, we observed that RcsB-dependent repression is required for the survival of bacteria when they are exposed to pancreatic fluids. We hypothesized that STM1485 plays an important role in Salmonella adaptation to pH changes, during transition in the gastrointestinal tract. We suggest that bacteria surviving the gastrointestinal environment invade the epithelial cells, where they can remain in vacuoles. In this new environment, acidity and magnesium starvation activate the expression of the RstA regulator in a PhoPQ-dependent manner, which in turn induces STM1485 expression. These levels of STM1485 allow increased bacterial replication within vacuoles to continue the course of infection.
Subject(s)
Acids/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial/drug effects , Salmonella typhimurium/metabolism , Transcription Factors/metabolism , Transcription, Genetic , Base Sequence , Gastrointestinal Tract/drug effects , Gastrointestinal Tract/microbiology , Microbial Viability , Promoter Regions, Genetic , Regulatory Elements, Transcriptional , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Signal Transduction , Transcription Factors/geneticsABSTRACT
BACKGROUND: Although the role of histamine H4 receptor (H4R) in immune cells is being extensively investigated, its immunomodulatory function in cancer is completely unknown. This study aimed to investigate the role of H4R in antitumour immunity in a model of triple-negative breast cancer. METHODS: We evaluated growth parameters, histological characteristics and the composition of tumour, splenic and tumour draining lymph node (TDLN) immune subsets, in a syngeneic model, developed orthotopically with 4T1 cells in H4R knockout (H4R-KO) and wild-type mice. RESULTS: Mice lacking H4R show reduced tumour size and weight, decreased number of lung metastases and percentage of CD4+ tumour-infiltrating T cells, while exhibiting increased infiltration of NK cells and CD19+ lymphocytes. Likewise, TDLN of H4R-KO mice show decreased CD4+ T cells and T regulatory cells (CD4+CD25+FoxP3+), and increased percentages of NK cells. Finally, H4R-deficient mice show decreased Tregs in spleens and non-draining lymph nodes, and a negative correlation between tumour weight and the percentages of CD4+, CD19+ and NK splenic cells, suggesting that H4R also regulates antitumour immunity at a systemic level. CONCLUSIONS: This is the first report that demonstrates the participation of H4R in antitumour immunity, suggesting that H4R could be a target for cancer treatment.
Subject(s)
Breast Neoplasms/genetics , Immunomodulation/genetics , Receptors, Histamine H4/genetics , T-Lymphocytes, Regulatory/immunology , Animals , Antigens, CD19/immunology , Breast Neoplasms/immunology , Breast Neoplasms/therapy , CD4-Positive T-Lymphocytes/immunology , Female , Forkhead Transcription Factors/immunology , Humans , Killer Cells, Natural/immunology , Mice , Mice, Knockout , Receptors, Histamine H4/immunologyABSTRACT
The Salmonella enterica serovar Typhimurium RcsCDB system regulates the synthesis of colanic acid and the flagellum as well as the expression of virulence genes. We previously demonstrated that the rcsC11 mutant, which constitutively activates the RcsB regulator, attenuates Salmonella virulence in an animal model. This attenuated phenotype was also produced by deletion of the slyA gene. In this work, we investigated if this antagonistic behavior is produced by modulating the expression of both regulator-encoding genes. We demonstrated that SlyA overproduction negatively regulates rcsB transcription. A bioinformatics analysis enabled us to identify putative SlyA binding sites on both promoters, P rcsDB and P rcsB , which control rcsB transcriptional levels. We also determined that SlyA is able to recognize and bind to these predicted sites to modulate the activity of both rcsB promoters. According to these results, SlyA represses rcsB transcription by direct binding to specific sites located on the rcsB promoters, thus accounting for the attenuated/virulence antagonistic behaviors. Moreover, we showed that the opposite effect between both regulators also physiologically affects the Salmonella motility phenotype. In this sense, we observed that under SlyA overproduction, P rcsB is repressed, and consequently, bacterial motility is increased. On the basis of these results, we suggest that during infection, the different RcsB levels produced act as a switch between the virulent and attenuated forms of Salmonella Thereby, we propose that higher concentrations of RcsB tilt the balance toward the attenuated form, while absence or low concentrations resulting from SlyA overproduction tilt the balance toward the virulent form.IMPORTANCE The antagonistic behavior of RcsB and SlyA on virulence gene expression led us to hypothesize that there is interplay between both regulators in a regulatory network and these could be considered coordinators of this process. Here, we report that the SlyA virulence factor influences motility behavior by controlling rcsB transcription from the P rcsB promoter. We also demonstrate that SlyA negatively affects the expression of the rcsB gene by direct binding to P rcsDB and P rcsB promoters. We suggest that different levels of RcsB act as a switch between the virulent and attenuated forms of Salmonella, where high concentrations of the regulator tend to tilt the balance toward the attenuated form and low concentrations or its absence tilt it toward the virulent form.
Subject(s)
Bacterial Proteins/biosynthesis , DNA-Binding Proteins/metabolism , Gene Expression Regulation, Bacterial , Repressor Proteins/metabolism , Salmonella typhimurium/genetics , Transcription Factors/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Binding Sites , Computational Biology , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , DNA-Binding Proteins/genetics , Flagella/physiology , Gene Expression , Locomotion , Promoter Regions, Genetic , Protein Binding , Repressor Proteins/genetics , Salmonella typhimurium/physiology , Transcription Factors/geneticsABSTRACT
RESUMEN Las aguas de desecho dispuestas en una corriente superficial, lagos, ríos, mar, sin ningún tratamiento, ocasionan graves inconvenientes de contaminación, que afectan la flora y la fauna. Estas aguas residuales, antes de ser vertidas en las masas receptoras, deben recibir un tratamiento adecuado, capaz de modificar sus condiciones físicas, químicas y microbiológicas, para evitar que su disposición cause la alteración y la degradación de los ecosistemas asociados y problemas de salud pública. La investigación tuvo como objetivo evaluar el efecto de mezclas de quitosano y extracto acuoso de la cáscara de naranja, a diferentes concentraciones, en el proceso de coagulación-floculación, de aguas residuales, para lo cual, se realizó una prueba de jarras con agitación rápida y lenta, para evaluar turbidez (NTU), demanda química de oxígeno (DQO), demanda bioquímica de oxígeno (DBO), sólidos suspendidos totales (SST) y SS (sólidos sedimentables). Los resultados mostraron que todos los tratamientos presentaron diferencias significativas (p<0,05), con la muestra control y fueron eficientes en la reducción de turbidez, DBO, DQO, SST, SS. El tratamiento que combinó quitosano y extracto acuoso de cáscara de naranja (50-50%), a un pH de 5,5, disminuyó significativamente (p<0,05) la turbidez, en 79%, demostrando, de manera preliminar, que el extracto acuoso de la cáscara de naranja acidifica la mezcla e incrementa la formación de flóculos aglomerados en una fase liviana de las muestras, convirtiéndose en un agente eficiente, para ser usado en el tratamiento de aguas residuales.
ABSTRACT Waste water disposed in a superficial current, lakes, rivers, the sea, without any treatment, causes great inconvenient pollution that affects the flora and fauna of a region. This wastewater, before being poured into the receiving body of water, must receive the appropriate treatment, in order to modify its physical, chemical and microbiological conditions, to avoid that its disposal causes the alteration and degradation of the associated ecosystems and problem with public health. This research had as an objective to assess the effect of chitosan and aqueous extract of orange peel shells at different concentrations on the coagulation-flocculation process of wastewater. In order to do this, a jar test with fast and slow pull was carried out, assessing turbidity (NTU), chemical oxygen demand (COD), biochemical oxygen demand (BOD), total suspended solids (TSS) and SS (solid sediments). The results showed that all the treatments presented significant differences (p<0.05) to the control sample, and were efficient in the decrease of NTU, DQO, DBO, SST, and SS. The treatment that combined chitosan and aqueous extract of orange peel (50-50%) at a pH or 5.5, decreased significantly (p<0.05) the turbidity in a 79%, showing in a preliminary way that the aqueous extract of orange peel shells acidifies the mix and increases the formation of agglomerated floccules in a light phase of the samples, becoming an efficient agent to be used in the treatment of wastewater.
ABSTRACT
Magnetohydrodynamics (MHD) is becoming more popular every day among developers of applications based on microfluidics, such as “lab on a chip” (LOC) and/or “micro-total analysis systems” (micro-TAS). Its physical properties enable fluid manipulation for tasks such as pumping, networking, propelling, stirring, mixing, and even cooling without the need for mechanical components, and its non-intrusive nature provides a solution to mechanical systems issues. However, these are not easy tasks. They all require precise flow control, which depends on several parameters, like microfluidics conductivity, the microfluidics conduit (channel) shape and size configuration, and the interaction between magnetic and electric fields. This results in a mathematical model that needs to be validated theoretically and experimentally. The present paper introduces the design of a 3D laminar flow involving an electrolyte in an annular open channel driven by a Lorentz force. For an organized description, first of all is provided an introduction to MHD applied in microfluidics, then an overall description of the proposed MHD microfluidic system is given, after that is focused in the theoretical validation of the mathematical model, next is described the experimental validation of the mathematical model using a customized vision system, and finally conclusions and future work are stated.