Ana Campa | Universidade de São Paulo (original) (raw)
Papers by Ana Campa
We studied here the effect of the acute phase protein serum amyloid A (SAA) on the oxidative burs... more We studied here the effect of the acute phase protein serum amyloid A (SAA) on the oxidative burst of neutrophils. Incubation of neutrophils with SAA increased the rate of oxygen uptake and the production of reactive oxygen species of neutrophils activated with opsonized zymosan (OZ). The increment in the neutrophil oxidative burst was dependent on SAA concentration in the range of 3^33 Wg protein ml 31 and was observed only in the presence of a relatively low amount of OZ (1U10 6 particles ml 31 ). SAA did not affect oxygen consumption and reactive oxygen production triggered by other stimuli, such as f-Met-Leu-Phe, phorbol myristate acetate or nonopsonized zymosan. Our finding points to a priming effect of SAA probably associated with mobilization of receptors for opsonized particles and strengthens the role of SAA as an effector of neutrophil functions in inflammation.
H In the presence of hydrogen peroxide, horseradish eroxidase (HRP) catalyzes the production of N... more H In the presence of hydrogen peroxide, horseradish eroxidase (HRP) catalyzes the production of Ncetyl-N-formyl-5-methoxykynuramine from melatoin. This reaction consumes oxygen and exhibits hemiluminescence in the 440–540 nm region. The exited cleavage product derived from the thermolysis f an intermediate dioxetane is suggested to be the mitting species. Chemiluminescence and the indole ing cleavage product were also observed when HRP/ 2O2 was replaced by phorbol myristate acetate or psonized zymosan-activated neutrophils. Azide, a yeloperoxidase inhibitor, strongly suppressed melaonin oxidation. Superoxide dismutase has a strong nhibitory effect on light emission but catalase and ric acid are without effect on the emission. The oxiation of melatonin by activated neutrophils may be elevant to the in vivo functions of myeloperoxidase nd melatonin. The possible biological implication of elatonin oxidation by neutrophils, especially in inammatory conditions, is discussed. © 2000 Acade...
Journal of Biological Methods
Research on melanogenesis, its regulation in health and disease, and the discovery of new molecul... more Research on melanogenesis, its regulation in health and disease, and the discovery of new molecules with pigmenting and depigmenting activities use different models. Here we standardize a protocol based on previous ones using primary human melanocytes and keratinocytes in co-cultures, in which melanogenesis was induced under mild conditions by the addition of tyrosine plus ammonium chloride (NH4Cl). The expression of MITF, TYR, TYRP1, and Melan-A as well as melanin content were measured. Furthermore, we extended this study to a reconstructed 3D model. Pigmentation was visually observable and melanosomes were identified by Fontana-Masson staining by the addition of tyrosine plus NH4Cl during the stratification phase. The 2D and 3D protocols proposed here circumvent limitations of previous models, using human primary cells and mild conditions for melanogenesis. These protocols offer a viable, robust, simple, and animal-free investigational option for human skin pigmentation studies an...
Cancer Cytopathology
Human papillomavirus (HPV) infection is the central factor for cervical cancer, whereas epithelia... more Human papillomavirus (HPV) infection is the central factor for cervical cancer, whereas epithelial immune mechanisms contribute to the progression of HPV infection and its associated lesions. The authors evaluated the expression of indoleamine 2,3‐dioxygenase (IDO) and tryptophan 2,3‐dioxygenase (TDO) in cervicovaginal samples from women with normal cervical epithelium or with different degrees of squamous intraepithelial lesions (SILs) and cervical cancer.
ABSTRACTAimIntense endotoxaemia and infection are able to reduce appetite and induce a catabolic ... more ABSTRACTAimIntense endotoxaemia and infection are able to reduce appetite and induce a catabolic state, therefore leading to weight loss. However, it is underexplored its late effects on energy homeostasis, regulation of body weight and glucose metabolism. Here we addressed whether serial intense endotoxaemia, characterized by an acute phase response and weight loss, could be an aggravating or predisposing factor to diet-induced obesity (DIO) and associated metabolic impairments.MethodsMale Swiss Webster mice were submitted to 8 consecutive doses of lipopolysaccharide (LPS - 10 mg/kg), followed by 10 weeks in high-fat diet (HFD).ResultsAfter the end of the acute endotoxaemia period, mice under chow diet recovered their weight rapidly, within one-week recovery period, which remained similar to its control counterparts. However, acute endotoxaemia caused a long-lasting adipose tissue expression of the inflammatory markers TLR-4, CD14 and serum amyloid A (SAA) and, when challenged by a...
Journal of neuro-oncology, Jan 11, 2017
Serum amyloid A1 (SAA1) is a sensitive acute phase reactant primarily produced by the liver in re... more Serum amyloid A1 (SAA1) is a sensitive acute phase reactant primarily produced by the liver in response to acute inflammation. We have recently shown that SAA affects proliferation, migration, and invasion of glioblastoma cell lines, which suggest its participation in the malignant process. Consistently, levels of SAA have been used as a non-invasive biomarker for the prognosis of many cancers. In this study, we aimed to investigate SAA serum levels and expression of SAA genes in human astrocytomas tissues. Serum and tissue samples were obtained from patients with astrocytoma grades I to III and glioblastoma (GBM or grade IV). Levels of circulating SAA were significantly higher in the serum of patients with AGII-IV when compared to non-neoplastic samples derived from non-neoplastic patients (NN) (p > 0.0001). Quantitative real time PCR (qRT-PCR) of 148 astrocytomas samples (grades I-IV) showed that SAA1 mRNA was significantly higher in GBM when compared to AGI-III and NN samples ...
Nutrients, Jan 21, 2017
Serum amyloid A (SAA) was recently associated with metabolic endotoxemia, obesity and insulin res... more Serum amyloid A (SAA) was recently associated with metabolic endotoxemia, obesity and insulin resistance. Concurrently, insufficient sleep adversely affects metabolic health and is an independent predisposing factor for obesity and insulin resistance. In this study we investigated whether sleep loss modulates SAA production. The serum SAA concentration increased in C57BL/6 mice subjected to sleep restriction (SR) for 15 days or to paradoxical sleep deprivation (PSD) for 72 h. Sleep restriction also induced the upregulation of Saa1.1/Saa2.1 mRNA levels in the liver and Saa3 mRNA levels in adipose tissue. SAA levels returned to the basal range after 24 h in paradoxical sleep rebound (PSR). Metabolic endotoxemia was also a finding in SR. Increased plasma levels of SAA were also observed in healthy human volunteers subjected to two nights of total sleep deprivation (Total SD), returning to basal levels after one night of recovery. The observed increase in SAA levels may be part of the i...
Molecular and Biological Physics of Living Systems, 1990
Since excited singlet species can be formed in biological systems (bioluminescence), the question... more Since excited singlet species can be formed in biological systems (bioluminescence), the question arises as to whether triplet species can also be formed biochemically. Electronically excited triplet species have an intrinsic lifetime which is much longer than that of singlet species, whereby they may potentially be much more useful. This raises a second question; if triplet species are formed, do they have a functional role? This question is of special importance since there are several photochemical-like processes which occur in vivo in the complete absence of light |1|. Substantial progress in the search for biochemically generated triplet species was made only as of the late seventies; the available |2–5| reviews contain references pertaining to a possible role of excited states in biological processes as considered by other authors.
Photochem Photobiol, 1989
Cellular immunology, Jan 26, 2015
Myeloperoxidase (MPO) is an important enzyme in the front-line protection against microorganisms.... more Myeloperoxidase (MPO) is an important enzyme in the front-line protection against microorganisms. In peripheral blood, it is accepted that MPO is only produced by myeloid-lineage cells. Thus, MPO presence is unexpected in lymphocytes. We showed recently that B1-lymphocytes from mice have MPO. Here, we showed that subsets of human peripheral B, CD4(+) and CD8(+) T lymphocytes express MPO. The content of MPO in lymphocytes was very low compared to neutrophils/monocytes with a preferential distribution in the nucleus and perinuclear region. Also, we performed a MPO mRNA expression analysis from human blood cells derived from microarray raw data publicly available, showing that MPO is modulated in infectious disease. MPO was increased in CD4(+) T lymphocytes from HIV chronic infection and in CD8(+) T lymphocytes from HCV-positive patients. Our study points out MPO as a multifunctional protein due to its subcellular localization and expression modulation in lymphocytes indicating alterna...
Physiological chemistry and physics and medical NMR
Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a m... more Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a membrane receptor-binding stimulus triggering an oxidative burst. One other considered sign of neutrophil activation is the appearance of a high-resolution 'H-NMR spectrum probably resulting from a rearrangement of neutral lipids in the membrane. The relationship between these two events is here studied. Neutrophils were recovered from a subcutaneous fungal infection using two murine lineages that respond differently to infection. It was concluded that rearrangement of lipids in the membrane, observable by NMR, appears also in neutrophils that do not exhibit the classical sign of activation, that is the increased expression of membrane receptors.
Role of Melatonin and Pineal Peptides in Neuroimmunomodulation, 1991
The Journal of rheumatology, 2007
To verify the importance of interleukin 18 (IL-18) in the pathogenesis of juvenile idiopathic art... more To verify the importance of interleukin 18 (IL-18) in the pathogenesis of juvenile idiopathic arthritis (JIA). We measured IL-18 levels in synovial fluid (SF) and serum, and determined their correlation with measures of disease activity and severity. Fifty patients with JIA (13 systemic, 13 polyarticular, 24 oligoarticular) and 25 matched controls were analyzed. Cytokine levels (IL-1beta, IL-1Ra, IL-6, and IL-18) were quantified in serum and SF by ELISA, and disease activity measures were evaluated immediately after knee articular puncture. Radiological assessment was made according to the Steinbrocker method. Statistical analysis was performed by Spearman's rank-order correlation and Mann-Whitney rank test. All the analyzed cytokine levels (IL-1, IL-1Ra, IL-6, and IL-18) were higher in patients' sera than in controls. Remarkably, in patients with JIA, IL-18 SF levels did not differ from those of serum; they were positively correlated. The levels of IL-18 (SF and serum) were...
Physiological chemistry and physics and medical NMR, 2001
Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a m... more Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a membrane receptor-binding stimulus triggering an oxidative burst. One other considered sign of neutrophil activation is the appearance of a high-resolution 'H-NMR spectrum probably resulting from a rearrangement of neutral lipids in the membrane. The relationship between these two events is here studied. Neutrophils were recovered from a subcutaneous fungal infection using two murine lineages that respond differently to infection. It was concluded that rearrangement of lipids in the membrane, observable by NMR, appears also in neutrophils that do not exhibit the classical sign of activation, that is the increased expression of membrane receptors.
Journal of leukocyte biology, 1996
The oxidative burst of polymorphonuclear neutrophils (PMN) and their ability to inhibit Paracocci... more The oxidative burst of polymorphonuclear neutrophils (PMN) and their ability to inhibit Paracoccidioides brasiliensis growth was studied in susceptible (B10.A) and resistant (A/J) mice. The cells were obtained after subcutaneous inoculation in air pouches, yielding highly pure PMN preparations; the number of cells was similar for both strains at 24 h and five times higher in the resistant strain at 15 days. The oxidative metabolism of these PMN was evaluated by the luminol and lucigen-enhanced chemiluminescence upon stimulation with PMA or killed P. brasiliensis (Pb). At 24 h of infection PMN from both strains showed similar responses. However, at 15 days a great enhancement of the Pb-stimulated luminol-enhanced chemiluminescence was observed only in PMN from resistant mice. Such increase was markedly inhibited by the addition of catalase. Independent of the mouse strain or time of infection of lucigen-enhanced chemiluminescence showed the same intensity. The lucigen-enhanced chemil...
We studied here the effect of the acute phase protein serum amyloid A (SAA) on the oxidative burs... more We studied here the effect of the acute phase protein serum amyloid A (SAA) on the oxidative burst of neutrophils. Incubation of neutrophils with SAA increased the rate of oxygen uptake and the production of reactive oxygen species of neutrophils activated with opsonized zymosan (OZ). The increment in the neutrophil oxidative burst was dependent on SAA concentration in the range of 3^33 Wg protein ml 31 and was observed only in the presence of a relatively low amount of OZ (1U10 6 particles ml 31 ). SAA did not affect oxygen consumption and reactive oxygen production triggered by other stimuli, such as f-Met-Leu-Phe, phorbol myristate acetate or nonopsonized zymosan. Our finding points to a priming effect of SAA probably associated with mobilization of receptors for opsonized particles and strengthens the role of SAA as an effector of neutrophil functions in inflammation.
H In the presence of hydrogen peroxide, horseradish eroxidase (HRP) catalyzes the production of N... more H In the presence of hydrogen peroxide, horseradish eroxidase (HRP) catalyzes the production of Ncetyl-N-formyl-5-methoxykynuramine from melatoin. This reaction consumes oxygen and exhibits hemiluminescence in the 440–540 nm region. The exited cleavage product derived from the thermolysis f an intermediate dioxetane is suggested to be the mitting species. Chemiluminescence and the indole ing cleavage product were also observed when HRP/ 2O2 was replaced by phorbol myristate acetate or psonized zymosan-activated neutrophils. Azide, a yeloperoxidase inhibitor, strongly suppressed melaonin oxidation. Superoxide dismutase has a strong nhibitory effect on light emission but catalase and ric acid are without effect on the emission. The oxiation of melatonin by activated neutrophils may be elevant to the in vivo functions of myeloperoxidase nd melatonin. The possible biological implication of elatonin oxidation by neutrophils, especially in inammatory conditions, is discussed. © 2000 Acade...
Journal of Biological Methods
Research on melanogenesis, its regulation in health and disease, and the discovery of new molecul... more Research on melanogenesis, its regulation in health and disease, and the discovery of new molecules with pigmenting and depigmenting activities use different models. Here we standardize a protocol based on previous ones using primary human melanocytes and keratinocytes in co-cultures, in which melanogenesis was induced under mild conditions by the addition of tyrosine plus ammonium chloride (NH4Cl). The expression of MITF, TYR, TYRP1, and Melan-A as well as melanin content were measured. Furthermore, we extended this study to a reconstructed 3D model. Pigmentation was visually observable and melanosomes were identified by Fontana-Masson staining by the addition of tyrosine plus NH4Cl during the stratification phase. The 2D and 3D protocols proposed here circumvent limitations of previous models, using human primary cells and mild conditions for melanogenesis. These protocols offer a viable, robust, simple, and animal-free investigational option for human skin pigmentation studies an...
Cancer Cytopathology
Human papillomavirus (HPV) infection is the central factor for cervical cancer, whereas epithelia... more Human papillomavirus (HPV) infection is the central factor for cervical cancer, whereas epithelial immune mechanisms contribute to the progression of HPV infection and its associated lesions. The authors evaluated the expression of indoleamine 2,3‐dioxygenase (IDO) and tryptophan 2,3‐dioxygenase (TDO) in cervicovaginal samples from women with normal cervical epithelium or with different degrees of squamous intraepithelial lesions (SILs) and cervical cancer.
ABSTRACTAimIntense endotoxaemia and infection are able to reduce appetite and induce a catabolic ... more ABSTRACTAimIntense endotoxaemia and infection are able to reduce appetite and induce a catabolic state, therefore leading to weight loss. However, it is underexplored its late effects on energy homeostasis, regulation of body weight and glucose metabolism. Here we addressed whether serial intense endotoxaemia, characterized by an acute phase response and weight loss, could be an aggravating or predisposing factor to diet-induced obesity (DIO) and associated metabolic impairments.MethodsMale Swiss Webster mice were submitted to 8 consecutive doses of lipopolysaccharide (LPS - 10 mg/kg), followed by 10 weeks in high-fat diet (HFD).ResultsAfter the end of the acute endotoxaemia period, mice under chow diet recovered their weight rapidly, within one-week recovery period, which remained similar to its control counterparts. However, acute endotoxaemia caused a long-lasting adipose tissue expression of the inflammatory markers TLR-4, CD14 and serum amyloid A (SAA) and, when challenged by a...
Journal of neuro-oncology, Jan 11, 2017
Serum amyloid A1 (SAA1) is a sensitive acute phase reactant primarily produced by the liver in re... more Serum amyloid A1 (SAA1) is a sensitive acute phase reactant primarily produced by the liver in response to acute inflammation. We have recently shown that SAA affects proliferation, migration, and invasion of glioblastoma cell lines, which suggest its participation in the malignant process. Consistently, levels of SAA have been used as a non-invasive biomarker for the prognosis of many cancers. In this study, we aimed to investigate SAA serum levels and expression of SAA genes in human astrocytomas tissues. Serum and tissue samples were obtained from patients with astrocytoma grades I to III and glioblastoma (GBM or grade IV). Levels of circulating SAA were significantly higher in the serum of patients with AGII-IV when compared to non-neoplastic samples derived from non-neoplastic patients (NN) (p > 0.0001). Quantitative real time PCR (qRT-PCR) of 148 astrocytomas samples (grades I-IV) showed that SAA1 mRNA was significantly higher in GBM when compared to AGI-III and NN samples ...
Nutrients, Jan 21, 2017
Serum amyloid A (SAA) was recently associated with metabolic endotoxemia, obesity and insulin res... more Serum amyloid A (SAA) was recently associated with metabolic endotoxemia, obesity and insulin resistance. Concurrently, insufficient sleep adversely affects metabolic health and is an independent predisposing factor for obesity and insulin resistance. In this study we investigated whether sleep loss modulates SAA production. The serum SAA concentration increased in C57BL/6 mice subjected to sleep restriction (SR) for 15 days or to paradoxical sleep deprivation (PSD) for 72 h. Sleep restriction also induced the upregulation of Saa1.1/Saa2.1 mRNA levels in the liver and Saa3 mRNA levels in adipose tissue. SAA levels returned to the basal range after 24 h in paradoxical sleep rebound (PSR). Metabolic endotoxemia was also a finding in SR. Increased plasma levels of SAA were also observed in healthy human volunteers subjected to two nights of total sleep deprivation (Total SD), returning to basal levels after one night of recovery. The observed increase in SAA levels may be part of the i...
Molecular and Biological Physics of Living Systems, 1990
Since excited singlet species can be formed in biological systems (bioluminescence), the question... more Since excited singlet species can be formed in biological systems (bioluminescence), the question arises as to whether triplet species can also be formed biochemically. Electronically excited triplet species have an intrinsic lifetime which is much longer than that of singlet species, whereby they may potentially be much more useful. This raises a second question; if triplet species are formed, do they have a functional role? This question is of special importance since there are several photochemical-like processes which occur in vivo in the complete absence of light |1|. Substantial progress in the search for biochemically generated triplet species was made only as of the late seventies; the available |2–5| reviews contain references pertaining to a possible role of excited states in biological processes as considered by other authors.
Photochem Photobiol, 1989
Cellular immunology, Jan 26, 2015
Myeloperoxidase (MPO) is an important enzyme in the front-line protection against microorganisms.... more Myeloperoxidase (MPO) is an important enzyme in the front-line protection against microorganisms. In peripheral blood, it is accepted that MPO is only produced by myeloid-lineage cells. Thus, MPO presence is unexpected in lymphocytes. We showed recently that B1-lymphocytes from mice have MPO. Here, we showed that subsets of human peripheral B, CD4(+) and CD8(+) T lymphocytes express MPO. The content of MPO in lymphocytes was very low compared to neutrophils/monocytes with a preferential distribution in the nucleus and perinuclear region. Also, we performed a MPO mRNA expression analysis from human blood cells derived from microarray raw data publicly available, showing that MPO is modulated in infectious disease. MPO was increased in CD4(+) T lymphocytes from HIV chronic infection and in CD8(+) T lymphocytes from HCV-positive patients. Our study points out MPO as a multifunctional protein due to its subcellular localization and expression modulation in lymphocytes indicating alterna...
Physiological chemistry and physics and medical NMR
Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a m... more Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a membrane receptor-binding stimulus triggering an oxidative burst. One other considered sign of neutrophil activation is the appearance of a high-resolution 'H-NMR spectrum probably resulting from a rearrangement of neutral lipids in the membrane. The relationship between these two events is here studied. Neutrophils were recovered from a subcutaneous fungal infection using two murine lineages that respond differently to infection. It was concluded that rearrangement of lipids in the membrane, observable by NMR, appears also in neutrophils that do not exhibit the classical sign of activation, that is the increased expression of membrane receptors.
Role of Melatonin and Pineal Peptides in Neuroimmunomodulation, 1991
The Journal of rheumatology, 2007
To verify the importance of interleukin 18 (IL-18) in the pathogenesis of juvenile idiopathic art... more To verify the importance of interleukin 18 (IL-18) in the pathogenesis of juvenile idiopathic arthritis (JIA). We measured IL-18 levels in synovial fluid (SF) and serum, and determined their correlation with measures of disease activity and severity. Fifty patients with JIA (13 systemic, 13 polyarticular, 24 oligoarticular) and 25 matched controls were analyzed. Cytokine levels (IL-1beta, IL-1Ra, IL-6, and IL-18) were quantified in serum and SF by ELISA, and disease activity measures were evaluated immediately after knee articular puncture. Radiological assessment was made according to the Steinbrocker method. Statistical analysis was performed by Spearman's rank-order correlation and Mann-Whitney rank test. All the analyzed cytokine levels (IL-1, IL-1Ra, IL-6, and IL-18) were higher in patients' sera than in controls. Remarkably, in patients with JIA, IL-18 SF levels did not differ from those of serum; they were positively correlated. The levels of IL-18 (SF and serum) were...
Physiological chemistry and physics and medical NMR, 2001
Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a m... more Neutrophils exposed to pro-inflammatory substances have an increased capability to respond to a membrane receptor-binding stimulus triggering an oxidative burst. One other considered sign of neutrophil activation is the appearance of a high-resolution 'H-NMR spectrum probably resulting from a rearrangement of neutral lipids in the membrane. The relationship between these two events is here studied. Neutrophils were recovered from a subcutaneous fungal infection using two murine lineages that respond differently to infection. It was concluded that rearrangement of lipids in the membrane, observable by NMR, appears also in neutrophils that do not exhibit the classical sign of activation, that is the increased expression of membrane receptors.
Journal of leukocyte biology, 1996
The oxidative burst of polymorphonuclear neutrophils (PMN) and their ability to inhibit Paracocci... more The oxidative burst of polymorphonuclear neutrophils (PMN) and their ability to inhibit Paracoccidioides brasiliensis growth was studied in susceptible (B10.A) and resistant (A/J) mice. The cells were obtained after subcutaneous inoculation in air pouches, yielding highly pure PMN preparations; the number of cells was similar for both strains at 24 h and five times higher in the resistant strain at 15 days. The oxidative metabolism of these PMN was evaluated by the luminol and lucigen-enhanced chemiluminescence upon stimulation with PMA or killed P. brasiliensis (Pb). At 24 h of infection PMN from both strains showed similar responses. However, at 15 days a great enhancement of the Pb-stimulated luminol-enhanced chemiluminescence was observed only in PMN from resistant mice. Such increase was markedly inhibited by the addition of catalase. Independent of the mouse strain or time of infection of lucigen-enhanced chemiluminescence showed the same intensity. The lucigen-enhanced chemil...