The receptor CXCR4 and its ligand CXCL12 play crucial roles in breast cancer. Despite the fact th... more The receptor CXCR4 and its ligand CXCL12 play crucial roles in breast cancer. Despite the fact that the spontaneous feline mammary carcinoma (FMC) is considered a suitable model for breast cancer studies, the importance of the CXCR4/CXCL12 axis in FMC is completely unknown. Therefore, this work aims to elucidate the role of CXCR4 and its ligand in the progression of FMC and metastatic disease. CXCR4 and CXCL12 expression was analyzed by immunohistochemistry and immunofluorescence on primary tumors (PT), regional and distant metastases of female cats with mammary carcinoma and correlated with serum CXCL12 levels, tumor molecular subtypes and clinicopathological features. CXCR4 was more expressed in PT than in metastases (p = 0.0067), whereas CXCL12 was highly expressed in metastatic lesions located in liver and lung (p < 0.0001), as reported for human breast cancer. Moreover, cats with CXCR4 positive PT exhibited significantly lower serum CXCL12 levels than cats with CXCR4 negativ...
The feline mammary carcinoma (FMC) is the third most common tumor in cat, sharing many clinicopat... more The feline mammary carcinoma (FMC) is the third most common tumor in cat, sharing many clinicopathological features with human breast cancer and thus, considered a suitable model for comparative oncology. Due to its poor prognosis, further studies are required to improve the diagnostic accuracy and treatment of cats with spontaneous mammary carcinoma. Recently, it was reported that the overexpression of stromal cell-derived factor-1 (SDF-1) has great value in human breast cancer diagnosis, suggesting that diagnostic tools and therapies targeting the SDF-1 ligand can improve the clinical outcome. In this study, we aimed to evaluate if serum SDF-1 levels can also be used as a biomarker of mammary carcinoma in cats and to analyze if serum SDF-1 levels are associated with clinicopathological features, linked to a specific FMC subtype or correlated with the tumor expression of SDF-1 receptor, the chemokine C-X-C motif receptor 4 (CXCR4). Results showed that cats with mammary carcinoma ha...
The monitoring of the microbial quality of fresh products in the industrial environment has mainl... more The monitoring of the microbial quality of fresh products in the industrial environment has mainly focused on bacterial indicators. Protozoa, such as Giardia duodenalis, Cryptosporidium spp., Toxoplasma gondii, and Cyclospora cayetanensis, are routinely excluded from detection and surveillance systems, despite guidelines and regulations that support the need for tracking and monitoring these pathogens in fresh food products. Previous studies performed by our laboratory, within the scope of the SafeConsume project, clearly indicated that consumption of fresh produce may be a source of T. gondii, thus posing a risk for the contraction of toxoplasmosis for susceptible consumers. Therefore, preliminary work was performed in order to assess the microbiological quality of vegetables, highlighting not only bacteria (Escherichia. coli, Listeria monocytogenes, and Salmonella spp.), but also the zoonotic protozoa G. duodenalis and Cryptosporidium spp. Although all samples were found to be acc...
Comparative Immunology, Microbiology and Infectious Diseases, 2015
The interaction between polymorphonuclear leukocytes (PMN) or neutrophils and Leishmania became a... more The interaction between polymorphonuclear leukocytes (PMN) or neutrophils and Leishmania became an interesting focus of research, since PMN turn out to be essential cells in transiently hosting the parasites. This study aims to evaluate whether L. infantum, the etiological agent of zoonotic visceral leishmaniasis, influences the in vitro functional activity of murine neutrophils. Phagocytosis, chemotaxis, oxidative burst, degranulation and apoptosis assays were performed. Cytokines, chemokines and toll-like receptors gene expression were evaluated by Real-time PCR. Results indicate that some of the innate features of PMN immunity were activated when in contact with L. infantum. However, parasites might negatively interfere with PMN defense mechanisms compromising the link between innate and acquired immunity. This work provides additional insights on the inflammatory immune interactions between neutrophils and L. infantum highlighting the role of PMN in Leishmania infection.
Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in... more Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in cellular and humoral immune responses in experimental leishmaniasis have been conducted. In this work, immunopathological changes induced by L. shawi in chronically infected BALB/c mice were investigated. Infected BALB/c mice developed increased lesion size associated with strong inflammatory infiltrate diffusely distributed in the dermis, with highly infected macrophages. The humoral immune response was predominantly directed toward the IgG1 isotype. The functional activity of CD4(+) and CD8(+) T cells showed significantly increased TNF-alpha mRNA levels associated with reduced IFN-gamma expression by CD4(+) T cells and the double negative (dn) CD4CD8 cell subset. High IL-4 levels expressed by CD8(+) T cells and dnCD4CD8 and TGF-beta by CD4(+) and CD8(+) T cells were detected, while IL-10 was highly expressed by all three cell subpopulations. Taken together, these results show an evident imbalance between TNF-alpha and IFN-gamma that is unfavorable to amastigote replication control. Furthermore, L. shawi seems to regulate different cell populations to express deactivating cytokines to avoid its own destruction. This study indicates BALB/c mice as a potentially good experimental model for further studies on American cutaneous leishmaniosis caused by L. shawi.
The human immune response to Plasmodium falciparum infection involves the release of cytokines th... more The human immune response to Plasmodium falciparum infection involves the release of cytokines that may contribute to the control of the parasites&amp;amp;amp;amp;amp;amp;#39; replication. These cytokines are also involved in the pathogenesis of the malaria caused by the infection, leading to the appearance of symptoms of varying severity. In a cross-sectional study, the expression of the genes that code for pro-inflammatory cytokines (tumour necrosis factor, interferon-gamma, interleukin-6 and interleukin-12) and anti-inflammatory cytokines (interleukin-10 and interleukin-4) among 80 children infected with P. falciparum (from a malaria-endemic area of Sudan) and five healthy controls (from a non-endemic area) was explored. The infected children were either non-sicklers, with severe malaria (18 children), mild malaria (30) or no symptoms of malaria (18), or asymptomatic sicklers (14). Interleukin-12 was found to be weakly expressed by all the groups of children. In general, compared with the other groups, the asymptomatic non-sicklers had lower expression of all the cytokines studied. The asymptomatic sicklers had significantly lower expression of tumour necrosis factor than the non-sicklers with severe malaria, but these two groups showed similar expression of interferon-gamma, interleukin-4 and interleukin-6. Gene expression of the regulatory cytokine, interleukin-10, by the asymptomatic sicklers was significantly lower than that by the non-sicklers with severe malaria but higher than that recorded in the non-sicklers with mild malaria. Their regulation of cytokine release appears to protect sicklers from clinical malaria.
Journal of controlled release : official journal of the Controlled Release Society, Jan 28, 2015
In the context of ocular pharmacology, there is a growing need for innovative delivery platforms ... more In the context of ocular pharmacology, there is a growing need for innovative delivery platforms for a convenient and sustained drug release into the eye, especially for chronic diseases that require the adoption of a strict insurmountable treatment regimen for a large part of the affected population, as in the case of glaucoma. Due to the large residence time of the contact lenses in the eye, its use for sustained drug delivery is quite promising. However, and despite the numerous therapeutic advantages arising from its use, the low affinity shown by most ophthalmic drugs for conventional contact lenses hinders the practical application of this technology. In this paper we elaborated a review of the various methods exploited so far to improve the contact lenses' characteristics as mechanisms for controlled and prolonged drug release for topical treatment of ocular diseases, with particular emphasis on the treatment of glaucoma.
ABSTRACT Leishmania chagasi is the aetiological agent of visceral leishmaniasis (VL). In the last... more ABSTRACT Leishmania chagasi is the aetiological agent of visceral leishmaniasis (VL). In the last years the role of neutrophils or polymorphonuclear cells (PMN) during Leishmania spp. infection has been the subject of great interest from the scientific community. This immune cell population is described to play protective functions but also with responsibility in disease progression. The main purpose of this work was to evaluate the role of PMN during the initial phase of experimental L. chagasi infection, by immunophenotyping different leukocyte subsets recruited to the site of parasite inoculation, quantification of chemokine and Toll-like receptors expression on the infection site and quantification of cytokine expression in internal organs after PMN depletion in C57BL/6 mice. It was observed that the absence of PMN did not interfere with chemokine and TLR-4 expression and leukocyte recruitment to the site of infection, although it seems to be a delay in parasite elimination. Nevertheless, Leishmania parasites spread to cervical lymph nodes. In these organs, PMN depletion influences the cytokine profile induced by the parasite in the beginning of infection (3 hours), leading to expression of pro and anti-inflammatory cytokines (IL1-β, TNF-α, TGF-β e IL-17). 144 hours post infection, the serine protease granzyme B production along with IFN-γ inhibition appear to be a reflection of the initial deficiency of PMN. With this work we provided evidence that PMN have a major role on cellular immunity during the early phases of L. chagasi infection that could be important for the development of alternative strategies for the control of VL.
In this study, protective effect and immune response elicited by protein fractions LiRic1 (> 7... more In this study, protective effect and immune response elicited by protein fractions LiRic1 (> 75kDa) and LiRic2 (< 37kDa) released by Leishmania infantum promastigotes were analysed in challenged BALB/c mice. Viable parasites were quantified in spleen and ...
The receptor CXCR4 and its ligand CXCL12 play crucial roles in breast cancer. Despite the fact th... more The receptor CXCR4 and its ligand CXCL12 play crucial roles in breast cancer. Despite the fact that the spontaneous feline mammary carcinoma (FMC) is considered a suitable model for breast cancer studies, the importance of the CXCR4/CXCL12 axis in FMC is completely unknown. Therefore, this work aims to elucidate the role of CXCR4 and its ligand in the progression of FMC and metastatic disease. CXCR4 and CXCL12 expression was analyzed by immunohistochemistry and immunofluorescence on primary tumors (PT), regional and distant metastases of female cats with mammary carcinoma and correlated with serum CXCL12 levels, tumor molecular subtypes and clinicopathological features. CXCR4 was more expressed in PT than in metastases (p = 0.0067), whereas CXCL12 was highly expressed in metastatic lesions located in liver and lung (p < 0.0001), as reported for human breast cancer. Moreover, cats with CXCR4 positive PT exhibited significantly lower serum CXCL12 levels than cats with CXCR4 negativ...
The feline mammary carcinoma (FMC) is the third most common tumor in cat, sharing many clinicopat... more The feline mammary carcinoma (FMC) is the third most common tumor in cat, sharing many clinicopathological features with human breast cancer and thus, considered a suitable model for comparative oncology. Due to its poor prognosis, further studies are required to improve the diagnostic accuracy and treatment of cats with spontaneous mammary carcinoma. Recently, it was reported that the overexpression of stromal cell-derived factor-1 (SDF-1) has great value in human breast cancer diagnosis, suggesting that diagnostic tools and therapies targeting the SDF-1 ligand can improve the clinical outcome. In this study, we aimed to evaluate if serum SDF-1 levels can also be used as a biomarker of mammary carcinoma in cats and to analyze if serum SDF-1 levels are associated with clinicopathological features, linked to a specific FMC subtype or correlated with the tumor expression of SDF-1 receptor, the chemokine C-X-C motif receptor 4 (CXCR4). Results showed that cats with mammary carcinoma ha...
The monitoring of the microbial quality of fresh products in the industrial environment has mainl... more The monitoring of the microbial quality of fresh products in the industrial environment has mainly focused on bacterial indicators. Protozoa, such as Giardia duodenalis, Cryptosporidium spp., Toxoplasma gondii, and Cyclospora cayetanensis, are routinely excluded from detection and surveillance systems, despite guidelines and regulations that support the need for tracking and monitoring these pathogens in fresh food products. Previous studies performed by our laboratory, within the scope of the SafeConsume project, clearly indicated that consumption of fresh produce may be a source of T. gondii, thus posing a risk for the contraction of toxoplasmosis for susceptible consumers. Therefore, preliminary work was performed in order to assess the microbiological quality of vegetables, highlighting not only bacteria (Escherichia. coli, Listeria monocytogenes, and Salmonella spp.), but also the zoonotic protozoa G. duodenalis and Cryptosporidium spp. Although all samples were found to be acc...
Comparative Immunology, Microbiology and Infectious Diseases, 2015
The interaction between polymorphonuclear leukocytes (PMN) or neutrophils and Leishmania became a... more The interaction between polymorphonuclear leukocytes (PMN) or neutrophils and Leishmania became an interesting focus of research, since PMN turn out to be essential cells in transiently hosting the parasites. This study aims to evaluate whether L. infantum, the etiological agent of zoonotic visceral leishmaniasis, influences the in vitro functional activity of murine neutrophils. Phagocytosis, chemotaxis, oxidative burst, degranulation and apoptosis assays were performed. Cytokines, chemokines and toll-like receptors gene expression were evaluated by Real-time PCR. Results indicate that some of the innate features of PMN immunity were activated when in contact with L. infantum. However, parasites might negatively interfere with PMN defense mechanisms compromising the link between innate and acquired immunity. This work provides additional insights on the inflammatory immune interactions between neutrophils and L. infantum highlighting the role of PMN in Leishmania infection.
Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in... more Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in cellular and humoral immune responses in experimental leishmaniasis have been conducted. In this work, immunopathological changes induced by L. shawi in chronically infected BALB/c mice were investigated. Infected BALB/c mice developed increased lesion size associated with strong inflammatory infiltrate diffusely distributed in the dermis, with highly infected macrophages. The humoral immune response was predominantly directed toward the IgG1 isotype. The functional activity of CD4(+) and CD8(+) T cells showed significantly increased TNF-alpha mRNA levels associated with reduced IFN-gamma expression by CD4(+) T cells and the double negative (dn) CD4CD8 cell subset. High IL-4 levels expressed by CD8(+) T cells and dnCD4CD8 and TGF-beta by CD4(+) and CD8(+) T cells were detected, while IL-10 was highly expressed by all three cell subpopulations. Taken together, these results show an evident imbalance between TNF-alpha and IFN-gamma that is unfavorable to amastigote replication control. Furthermore, L. shawi seems to regulate different cell populations to express deactivating cytokines to avoid its own destruction. This study indicates BALB/c mice as a potentially good experimental model for further studies on American cutaneous leishmaniosis caused by L. shawi.
The human immune response to Plasmodium falciparum infection involves the release of cytokines th... more The human immune response to Plasmodium falciparum infection involves the release of cytokines that may contribute to the control of the parasites&amp;amp;amp;amp;amp;amp;#39; replication. These cytokines are also involved in the pathogenesis of the malaria caused by the infection, leading to the appearance of symptoms of varying severity. In a cross-sectional study, the expression of the genes that code for pro-inflammatory cytokines (tumour necrosis factor, interferon-gamma, interleukin-6 and interleukin-12) and anti-inflammatory cytokines (interleukin-10 and interleukin-4) among 80 children infected with P. falciparum (from a malaria-endemic area of Sudan) and five healthy controls (from a non-endemic area) was explored. The infected children were either non-sicklers, with severe malaria (18 children), mild malaria (30) or no symptoms of malaria (18), or asymptomatic sicklers (14). Interleukin-12 was found to be weakly expressed by all the groups of children. In general, compared with the other groups, the asymptomatic non-sicklers had lower expression of all the cytokines studied. The asymptomatic sicklers had significantly lower expression of tumour necrosis factor than the non-sicklers with severe malaria, but these two groups showed similar expression of interferon-gamma, interleukin-4 and interleukin-6. Gene expression of the regulatory cytokine, interleukin-10, by the asymptomatic sicklers was significantly lower than that by the non-sicklers with severe malaria but higher than that recorded in the non-sicklers with mild malaria. Their regulation of cytokine release appears to protect sicklers from clinical malaria.
Journal of controlled release : official journal of the Controlled Release Society, Jan 28, 2015
In the context of ocular pharmacology, there is a growing need for innovative delivery platforms ... more In the context of ocular pharmacology, there is a growing need for innovative delivery platforms for a convenient and sustained drug release into the eye, especially for chronic diseases that require the adoption of a strict insurmountable treatment regimen for a large part of the affected population, as in the case of glaucoma. Due to the large residence time of the contact lenses in the eye, its use for sustained drug delivery is quite promising. However, and despite the numerous therapeutic advantages arising from its use, the low affinity shown by most ophthalmic drugs for conventional contact lenses hinders the practical application of this technology. In this paper we elaborated a review of the various methods exploited so far to improve the contact lenses' characteristics as mechanisms for controlled and prolonged drug release for topical treatment of ocular diseases, with particular emphasis on the treatment of glaucoma.
ABSTRACT Leishmania chagasi is the aetiological agent of visceral leishmaniasis (VL). In the last... more ABSTRACT Leishmania chagasi is the aetiological agent of visceral leishmaniasis (VL). In the last years the role of neutrophils or polymorphonuclear cells (PMN) during Leishmania spp. infection has been the subject of great interest from the scientific community. This immune cell population is described to play protective functions but also with responsibility in disease progression. The main purpose of this work was to evaluate the role of PMN during the initial phase of experimental L. chagasi infection, by immunophenotyping different leukocyte subsets recruited to the site of parasite inoculation, quantification of chemokine and Toll-like receptors expression on the infection site and quantification of cytokine expression in internal organs after PMN depletion in C57BL/6 mice. It was observed that the absence of PMN did not interfere with chemokine and TLR-4 expression and leukocyte recruitment to the site of infection, although it seems to be a delay in parasite elimination. Nevertheless, Leishmania parasites spread to cervical lymph nodes. In these organs, PMN depletion influences the cytokine profile induced by the parasite in the beginning of infection (3 hours), leading to expression of pro and anti-inflammatory cytokines (IL1-β, TNF-α, TGF-β e IL-17). 144 hours post infection, the serine protease granzyme B production along with IFN-γ inhibition appear to be a reflection of the initial deficiency of PMN. With this work we provided evidence that PMN have a major role on cellular immunity during the early phases of L. chagasi infection that could be important for the development of alternative strategies for the control of VL.
In this study, protective effect and immune response elicited by protein fractions LiRic1 (> 7... more In this study, protective effect and immune response elicited by protein fractions LiRic1 (> 75kDa) and LiRic2 (< 37kDa) released by Leishmania infantum promastigotes were analysed in challenged BALB/c mice. Viable parasites were quantified in spleen and ...
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Papers by Claudia S Marques