Enhanced apoptotic index in hepatocytes, Kupffer cells, and inflammatory infiltrate showed positive correlation with hepatic lesion intensity, parasite load, and clinical status in naturally Leishmania-infected dogs.

It is unknown if Leishmania amastigote infections affect hepatocytes and Kupffer cell apoptosis, and the role played by apoptosis in liver lesions in leishmaniasis is still unclear. Clinically affected and subclinically infected dogs with leishmaniosis and uninfected controls were assessed. Parasite load, biochemical markers for evaluation of liver damage, morphometry (area, perimeter, number of inflammatory focus, major and minor diameters), apoptosis in hepatic tissue (hepatocytes, Kupffer cells, and inflammatory infiltrates) and cellularity in inflammatory foci were quantified. The parasite load in clinically affected dogs proved to be higher than in the other groups. All morphometric parameters (area, perimeter, number of inflammatory focus, major and minor diameters) from clinically affected were higher than the values found in the subclinically infected and uninfected control dogs. Only clinically affected dogs presented high levels of ALT, FA, GGT and cholesterol in serum. Strong positive correlation was observed between biochemical markers for evaluation of liver damage (ALT, FA, GGT and cholesterol) and hepatic apoptosis (hepatocytes, Kupffer cells, and inflammation). Clinically affected dogs showed a more intense hepatic lesion. Hepatocytes showed a higher rate of apoptosis in Leishmania-infected dogs than in uninfected control dogs. The Kupffer cell apoptotic index and apoptosis within the inflammatory infiltrates were higher in clinically affected dogs. The apoptotic index evaluated in hepatocytes, Kupffer cells, and inflammatory infiltrates showed a positive correlation with the intensity of the hepatic lesion, parasite load, and clinical status. Apoptotic cells also showed positive immunostaining for TUNEL, Bcl2, and Bax. Our data showed that hepatic apoptosis was related to the severity of liver damage, the progression of infection, and the parasite load in leishmaniasis. Apoptotic regulated cell recruitment modulated the inflammatory response and favored the survival and dissemination of parasites, depending on the clinical status of the Leishmania-infected dogs.

Enhanced activation of eosinophils in peripheral blood and implications for eosinophilic esophagitis diagnosis.

BACKGROUND AND AIM: Eosinophils are markers of the eosinophilic esophagitis (EoE) disease, and this work aimed to assess whether activation of eosinophils could be a noninvasive test to contribute for EoE diagnosis. METHODS: The activation state of peripheral blood eosinophils in EoE patients and control subjects was assessed based on the morphological aspects of the eosinophil after adherence to slide. Cyclooxygenase-2 and 5-lipoxygenase expressions were evaluated by means of immunofluorescence microscopy to verify if and which eicosanoid pathway is triggered in eosinophils in blood in EoE. RESULTS: The eosinophils of patients with EoE were significantly more activated than those of control individuals. The lowest percentage of normal eosinophils for control subjects was 40%, while the highest percentage of eosinophils of normal aspect for patients with EoE was 32%. Considering 36% as a cutoff for normal eosinophils, this value differentiated all individuals with EoE from individuals without the disease with a sensitivity of 100%, considering the diagnosis of EoE as currently defined. Eosinophils of EoE patients showed higher expression of cyclooxygenase-2 than those of control subjects. CONCLUSIONS: The quantification of morphological changes in eosinophils is a feasible, easy, and reliable manner to identify EoE patients. Therefore, patients with symptoms of esophageal dysfunction showing higher than 36% activated eosinophils in peripheral blood could be a useful way to help definition and diagnostic criterion for EoE.

The effects of EPA and DHA enriched fish oil on nutritional and immunological markers of treatment naïve breast cancer patients: a randomized double-blind controlled trial.

BACKGROUND: We evaluated the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids enriched fish oil (FO) on nutritional and immunological parameters of treatment naïve breast cancer patients. METHODS: In a randomized double blind controlled trial, the FO group (FG) patients were supplemented with 2 g/ day of FO concentrate containing 1.8 g of n-3 fatty acids during 30 days. The placebo group (PG) received 2 g/ day of mineral oil. At baseline and after the intervention, plasma levels of n-3 fatty acids, dietary intake, weight, body composition, biochemical and immunological markers were assessed. RESULTS: At the end of the intervention period, no between group differences were observed regarding anthropometric parameters. There was a significant increase in the plasma phospholipid EPA (p = 0.004), DHA (p = 0.007) of the FG patients. In FG patients the percentages of peripheral blood CD4+ T lymphocytes and serum high sensitivity C-reactive protein (hsCRP) levels were maintained while in PG patients there was a significant increase in hsCRP (p = 0.024). We also observed a significant reduction in the percentage of CD4+ T lymphocytes in the peripheral blood (p = 0.042) of PG patients. No changes in serum proinflammatory cytokine and prostaglandin E2 levels were observed. CONCLUSIONS: Supplementation of newly diagnosed breast cancer patients with EPA and DHA led to a significant change in the composition of plasma fatty acids, maintained the level of CD4+ T cells and serum levels of hsCRP, suggestive of a beneficial effect on the immune system and less active inflammatory response. TRIAL REGISTRATION: Brazilian Clinical Trials Registry (REBEC): RBR-2b2hqh. Registered 29 April 2013, retrospectively registered.

Enhanced neutrophil phagocytic capacity in rheumatoid arthritis related to the autoantibodies rheumatoid factor and anti-cyclic citrullinated peptides.

BACKGROUND: There is no consensus on the mechanisms by which anti-cyclic citrullinated peptide antibodies (anti-CCP) and rheumatoid factor (RF) influence the pathogenesis of rheumatoid arthritis (RA). The current study verified if the presence of RF or anti-CCP is associated with phagocytic capacity and reactive oxygen species (ROS) production by phagocytes in RA patients to better clarify the role played by these antibodies in pathogenesis of the disease. METHODS: A cohort of 30 RA patients followed from early stages of the disease were characterized by positivity for RF or anti-CCP, disease activity score (DAS-28), health assessment questionnaire (HAQ), use of synthetic or biologic therapy, lifestyle, comorbidities and radiographic erosions. Phagocytic capacity against Saccharomyces cerevisiae and superoxide anion production were assessed in RA patients and compared with 20 healthy controls. Phagocytic capacity and superoxide anion production were also compared between RF- and anti-CCP-positive and -negative RA patients. RESULTS: Anti-CCP- and RF-positive RA patients had higher neutrophil phagocytic capacity than anti-CCP- (p = 0.005) and RF (p = 0.005)-negative individuals through pattern-recognition receptors. As assessed via pattern recognition or opsonin receptors, neutrophils and monocytes from RA patients presented overall higher phagocytic capacity than neutrophils and monocytes from healthy controls (p < 0.05). Furthermore, RA patients also showed a higher capacity for producing cytotoxic oxygen radicals (p = 0.0026). Phagocytosis and superoxide anion production did not correlate with any of the clinical variables analyzed in this study. CONCLUSIONS: This study showed increased phagocytosis by neutrophils in RA patients who were positive for anti-CCP and RF autoantibodies. Furthermore, there was an overall hyperactivation of the phagocytes in RA patients. Our data suggest that anti-CCP and RF may indirectly enhance the inflammation cascade involving neutrophils and may indirectly sustain tissue damage in RA. Targeting the production of these autoantibodies may be a promising strategy in the management of RA.

[Epidemiological situation of malaria in the brazilian amazon region, 2003 to 2012]. / Situação epidemiológica da malária na região amazônica brasileira, 2003 a 2012.

OBJECTIVE: To describe the epidemiological status of malaria in the Brazilian Amazon region between 2003 and 2012. METHODS: The present retrospective ecological study employed data from the Brazilian Epidemiological Surveillance and Malaria Communication System (SIVEP-Malária/SVS/MS), Hospital Admissions System (SIH/DATASUS/MS), and Mortality Information System (SIM). For each year, the percentage of Plasmodium falciparum cases, the number of admissions, and deaths and lethality due to malaria were determined. The distribution of P. falciparum cases in each state was also described. Data from 2012 were compared to data from 2005, when the Amazon region recorded a peak number of cases, and with data from 2011. RESULTS: In 2012, 241 806 malaria cases were recorded in the region, a reduction of 60.1% vs. 2005 and of 9.1% vs. 2011. Between 2003 and 2005, there was an increase of 48.3% in the number of cases, with 606 069 recorded cases in 2005. Since 2006, a declining trend in number of cases has been observed, especially for P. falciparum, with 155 169 cases notified in 2005 vs. 35 385 in 2012 (reduction of 77.2%). Between 2005 and 2012, the number of malaria hospital admissions (74,6%) and deaths (54,4%) was also reduced. CONCLUSIONS: Despite a decline in the number of malaria cases, the possible emergence of drug-resistant parasites and the lower frequency of P. falciparum indicate the need to adopt new surveillance strategies, more sensitive tools, and integrated vector management to achive a bold, but not impossible, goal: the elimination of P. falciparum.

Pravastatin modulates macrophage functions of Leishmania (L.) amazonensis-infected BALB/c mice.

The control of leishmaniases poses an important challenge due to the scarcity and toxicity of the pharmacological options available. We have previously shown that pravastatin significantly improves the course of the disease in Leishmania (L.) amazonensis-infected BALB/c mice. Since the drug caused no direct effect on the parasite, we decided to evaluate its immunomodulatory action in this experimental model. To evaluate the impact of pravastatin treatment, BALB/c mice infected or not with L. (L.) amazonensis were treated with pravastatin (20 mg/kg daily) or saline during 30 or 90 days and phagocytosis, hydrogen peroxide, nitric oxide and the tumor necrosis factor production by peritoneal macrophages were assessed. We showed that pravastatin increased the phagocytosis mediated by complement and immunoglobulin receptors (63.5 to 130.3; p=0.03, t test), but not that occurring via pattern recognition receptors, induced a rise of nitric oxide production by macrophages (2.1 µM to 12.9 µM; p=0.04, Mann-Whitney test), endowing these cells to better kill ingested leishmania organisms, caused no modification of the otherwise increased production of hydrogen peroxide by macrophages, and reduced the overproduction of tumor necrosis factor (166.6 pg/mL to 3.9 pg/mL; p=0.016, Mann-Whitney test), a major component of the exacerbated inflammation associated to leishmaniases. Our findings point to the potential usefulness of pravastatin as an adjunct to the treatment of leishmaniases, based on its powerful immunomodulatory effects and low toxicity.

MCP-1/IL-12 ratio expressions correlated with adventitial collagen depositions in renal vessels and IL-4/IFN-γ expression correlated with interstitial collagen depositions in the kidneys of dogs with canine leishmaniasis.

Collagen deposition is a common event in chronic inflammation, and canine Leishmaniosis (CanL) is generally associated with a long and chronic evolution. Considering that the kidney shows fibrinogenic changes during CanL, and the balance of cytokines/chemokines regulates the profibrinogenic and antifibrinogenic immune responses differently, it can be hypothesized that the balance of cytokines/chemokines can be differentially expressed in the renal tissue in order to determine the expression of collagen depositions in the kidneys. This study aimed to measure collagen deposition and to evaluate cytokine/chemokine expressions in the kidney by means of qRT-PCR in sixteen Leishmania-infected dogs and six uninfected controls. Kidney fragments were stained with hematoxylin & eosin (H&E), Masson's Trichrome, Picrosirius Red, and Gomori's reticulin. Intertubular and adventitial collagen depositions were evaluated by the morphometric approach. Cytokine RNA expressions were measured by means of qRT-PCR to identify molecules involved in chronic collagen depositions in kidneys with CanL. Collagen depositions were related to the presence of clinical signs, and more intense intertubular collagen depositions occurred in infected dogs. Adventitial collagen deposition, as morphometrically measured by the average area of the collagen, was more intense in clinically affected dogs than in subclinically infected dogs. TNF-α/TGF-ß, MCP1/IL-12, CCL5/IL-12, IL-4/IFN-γ, and IL-12/TGF-ß expressions were associated with clinical manifestations in dogs with CanL. The IL-4/IFN-α ratio was more commonly expressed and upregulated in clinically affected dogs, and downregulated in subclinically infected dogs. Furthermore, MCP-1/IL-12 and CCL5/IL-12 were more commonly expressed in subclinically infected dogs. Strong positive correlations were detected between morphometric values of interstitial collagen depositions and MCP-1/IL-12, IL-12, and IL-4 mRNA expression levels in the renal tissues. Adventitial collagen deposition was correlated with TGF-ß, IL-4/IFN-γ, and TNF-α/TGF-ß. In conclusion, our results showed the association of MCP-1/IL-12 and CCL5/IL-12 ratios with an absence of clinical signs, as well as an IL-4/IFN-α ratio with adventitial and intertubular collagen depositions in dogs with visceral leishmaniosis.

Association between human papillomavirus infection and cytological abnormalities during early follow-up of invasive cervical cancer.

Considering the limitations of cytology for detection of residual cervical cancer after radiotherapy, the aim of this study was to evaluate the frequency and viral load of high-risk HPV in cervical and vaginal samples during the early follow-up of patients treated for invasive cervical cancer and correlate the results with cytological examination. Conventional cytology and hybrid capture test were performed on cervical and vaginal samples of 52 women with invasive cervical carcinoma 3 months after therapy. High-risk HPV was detected in 46.1% of the samples and the median and the range of the ratio relative light unit (RLU)/cutoff (CO) (estimated viral load) in positive samples was 1.71 (1-2120.03). No significant difference was observed in viral frequency and in median of the ratio RLU/CO between samples of patients at different stages (I-III) and between cervical samples, from patients treated by exclusive radiotherapy with or without chemotherapy, and vaginal samples, from patients who underwent to hysterectomy and radiotherapy with or without chemotherapy. Cytological abnormalities were more frequent significantly in samples with HPV than in samples without HPV. The viral load was also higher significantly in samples with cytological abnormalities when compared with the samples without cytological abnormalities. In conclusion, HPV detection methods may be useful during the early follow-up as a complement to conventional cytology for the diagnosis of residual cervical cancer after radiotherapy.

Effects of periodontal therapy on phagocytic activity of peripheral blood neutrophils - evidence for an extrinsic cellular defect.

PURPOSE: To compare phagocytic activity of peripheral blood neutrophils from subjects with and without periodontal disease and evaluate the effects of periodontal therapy in individuals with similar levels of resolution of inflammation at the end of treatment. MATERIALS AND METHODS: To compare the phagocytic activity of neutrophils, peripheral blood was collected from 27 control subjects with a healthy periodontium and 28 periodontitis subjects before and after treatment. The phagocytosis of killed Saccharomyces cerevisiae, pre-sensitised or non-sensitised with fresh serum from the donor, was quantified and a phagocytic index was calculated as the mean number of yeast cells phagocytised by the percentage of neutrophils involved in phagocytosis. RESULTS: Prior to periodontal treatment, subjects with periodontitis exhibited significantly lower neutrophil phagocytic activity than control subjects with a healthy periodontium. Periodontal treatment significantly improved in clinical periodontal status and resulted in significantly increased phagocytosis of both pre-sensitised (from 113.0 pre- to 157.0 post-treatment, P = 0.02) and non-sensitised S. cerevisiae (from 1.5 pre- to 3.5 post-treatment, P = 0.001), to levels observed in control subjects. CONCLUSIONS: The phagocytic activity of peripheral blood neutrophils from subjects with periodontal disease was lower than that of healthy controls. Subjects who underwent non-surgical periodontal treatment and strict supportive therapy for 6 months showed improved phagocytic activity in peripheral blood neutrophils. The phagocytic index values from subjects with periodontal disease after treatment achieved those found in the control group.

Nucleolar organizer region proteins enhancement in nucleoplasm's of renal tubular cells is an indication of kidney impairment in Leishmania-infected dogs.

Hyperactivation of tubular cells contributes for the progression of kidney lesions. The exacerbated expression of immunological proteins and ribosomal DNA (rDNA) transcriptional activity are observed in tubular cells. This intensified expression results in more prominent hypertrophic changes and is often accompanied by increased expression of factors involved in different phases of ribosomal biosynthesis, such as the nucleolar organizer regions (NOR). The aim of this study was to evaluate whether there is an association between NOR proteins, renal impairment, and clinical status in Leishmania-infected dogs (CanL). Forty-five dogs with CanL and six uninfected controls were assessed in this study. PCR was performed to detect parasites' nucleic acids in kidney. Histopathological analyses were performed in kidney fragments, and NOR was detected by Ag stain (AgNOR). Leishmania-infected dogs showed more intense inflammation and collagen deposition compared with uninfected controls. Biochemical alterations were observed only in Leishmania-infected dogs. AgNORs per cell were significantly higher in clinically affected dogs and higher histopathological lesion score was observed in Leishmania-infected dogs. Positive correlations between number of NORs per cell in medullary region and histopathological lesion score were observed. Furthermore, AgNOR expression, intensity of renal lesions, and clinical sigs was associated in Leishmania-infected dogs. We propose that the detection of AgNOR proteins could be used to better estimate the kidney tubular damage at the time of examination in Leishmania-infected dogs as a marker to estimate renal impairment in dogs with CanL.

Influence of long-term treatment with pravastatin on the survival, evolution of cutaneous lesion and weight of animals infected by Leishmania amazonensis.

The high toxicity of current drugs for treatment of leishmaniasis is a major hindrance for controlling the disease. Pravastatin is a well-known drug with anti-inflammatory and immunomodulatory properties that may modulate host defense mechanisms against Leishmania. We evaluated the influence of prolonged pravastatin treatment on the survival of Leishmania amazonensis-infected animals (BALB/c, C57BL6 mice and Syrian hamsters), including weekly measurement of cutaneous lesions (footpad thickness) and weight. Pravastatin improved survival of Leishmania-infected BALB/c mice but not of infected C57BL6 mice or hamsters. On the 50th week of follow-up, 71% of pravastatin-treated Leishmania-infected BALB/c mice were alive against 29% of control group (p<0.01). Low footpad thickness was found on BALB/c pravastatin treated mice from the 14th week (p<0.05), and 20th week onward for C57BL6 treated mice. Pravastatin treatment decreased weight loss in Leishmania-infected C57BL6 mice and Syrian hamsters, but not infected BALB/c mice. Our results points to beneficial effects of pravastatin on the evolution of the disease in the murine leishmaniasis model.

Serum fatty acid synthase levels and n-3 fatty acid intake in patients with breast cancer.

PURPOSE: Fatty acid synthase (FASN) is a key enzyme in fatty acid biosynthesis that is usually over-expressed in patients with breast cancer, but its relationship with the patient's dietary habit is not clear. A higher intake of n-3 polyunsaturated fatty acids is related to reduced breast carcinogenesis in vitro and in vivo. The aim of this study was to clinically investigate the association between serum FASN levels and fatty acid intake in women newly diagnosed with breast cancer. METHODS: In a case-control cross-sectional study, with 18 breast cancer patients and 29 controls, we evaluated nutritional status, dietary intake, and serum FASN levels. Statistical analyses were carried out with parametric and non-parametric tests, according to the sample's normality distribution. RESULTS: The mean age of breast cancer group (n = 18) and control group (n = 29) was 46.8 ± 9.7 y and 44.4. ± 8.6 y, respectively. Mean serum concentration of FASN in breast cancer group was significantly higher (132.51 ± 95.05 ng/mL) than in control group (36.88 ± 20.87 ng/mL) (p < 0.0001). Among breast cancer group, serum FASN levels of premenopausal women were significantly higher than those of postmenopausal women (p = 0.026). There was no significant difference between the early and late disease stages in regard to serum FASN levels in breast cancer group. Mean nutrient intake was similar and n-3 docosahexaenoic acid intake was low in both groups. We observed no association regarding fatty acid intake and serum FASN levels. CONCLUSION: These data suggest that dietary n-3 fatty acid has no association with serum FASN levels among newly diagnosed breast cancer patients.

The balance between IL-12/IL4 in renal tissue switches the inflammatory response arm and shows relationship with the clinical signs in Leishmania-infected dogs.

The pathogenesis of Canine leishmaniosis (CanL) is associated with altered cytokine expression and parasitic tissue shows a lot of inflammation. The aim of this study was to assess the renal inflammation and cytokine expression in eight symptomatic and eight asymptomatic Leishmania- infected dogs, and seven uninfected control dogs. Kidney fragments were stained with hematoxylin and eosin for morphometric evaluation. mRNA expression levels of interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), interleukin (IL)-2, IL-4, IL-10, and IL-12 were assessed in the kidney fragments using quantitative real time-polymerase chain reaction. Inflammation, quantified by the average area of the infiltrated immune cells, was greater in symptomatic dogs than in those asymptomatic, whereas asymptomatic dogs exhibited higher inflammation than the control dogs (p > 0.05, Tukey's test). Expression levels of IFN-γ, TNF-α, IL-4, IL-10, and IL-12 were upregulated in symptomatic dogs and downregulated in asymptomatic dogs compared with those of the uninfected group. Furthermore, IL-4 showed higher expression in symptomatic dogs than in asymptomatic ones (p < 0.05, Mann-Whitney test), which was directly associated with clinical manifestations (p < 0.05, Chi-square test). However, IL-12 was predominantly expressed in symptomatic dogs, shifting the balance from IL-12/IL-4 to IL-12, which elicits a change in the inflammatory response. Leishmania was not found in the renal tissues in any one of the studied groups. Our data suggests that the balance between IL-12 and IL-4 plays an important role in the regulation of inflammation in renal tissue and clinical presentations in CanL.

Enhanced apoptotic index, chemokines and inflammatory recruitment in renal tissues shows relationship with the clinical signs in Leishmania-infected dogs.

Apoptosis is associated with resolution of inflammation. However, apoptosis may also occur in active inflammation, balancing inflammatory recruitment instead of a resolution event. To test that hypothesis, we measured apoptosis and chemokines expression, involved in recruitment of inflammatory cells. Clinical affected and subclinically infected dogs with canine leishmaniosis (CanL) and uninfected controls were assessed. Apoptosis in renal tissue (glomeruli, tubules, and inflammatory infiltrate) and cellularity in inflammatory foci were quantified. Messenger RNA of CCL5, CCL4, MCP-1, MCP-2, Caspase (Casp) 3, Casp 8, Casp 9, Bax, Bcl2 and Fas were quantified by qRT PCR. Clinical affected dogs showed more intense inflammation and higher cellularity in the inflammatory infiltrates than subclinically infected ones, which were higher than controls. Glomerular and tubular cells showed higher apoptotic index in clinical affected dogs when compared to controls. Apoptosis within the inflammatory infiltrates was higher in clinical affected dogs. Bax/Bcl2 ratio and CCL4 showed higher expression in kidney from clinical affected when compared to subclinically infected dogs. Casp 3/CCL4 ratio expression were higher in subclinically infected dogs than in the clinical affected group. Additionally, results suggest that Casp 3/CCL4 ratio is balancing towards an inflammatory recruitment and CCL4 and Bax/Bcl2 ratio expression is associated with active inflammation in clinical affected CanL. Data demonstrate that apoptosis was not always correlated with resolution of inflammation, when a morphometric and a molecular evaluation were performed concomitantly. In kidneys of Leishmania infected dogs, apoptosis and chemokines may be balancing inflammatory recruitment. In conclusion, Bax/Bcl2 ratio, chemokines, Casp 8, Casp 3 and Fas were associated with renal apoptosis, active inflammation and increased inflammatory recruitment observed in clinical affected animals, influencing the clinical presentation of leishmaniosis.

Phylloseptin-1 is Leishmanicidal for Amastigotes of Leishmaniaamazonensis Inside Infected Macrophages.

Leishmania protozoans are the causal agents of neglected diseases that represent an important public health issue worldwide. The growing occurrence of drug-resistant strains of Leishmania and severe side effects of available treatments represent an important challenge for the leishmaniases treatment. We have previously reported the leishmanicidal activity of phylloseptin-1 (PSN-1), a peptide found in the skin secretion of Phyllomedusaazurea (=Pithecopus azureus), against Leishmaniaamazonensis promastigotes. However, its impact on the amastigote form of L. amazonensis and its impact on infected macrophages are unknown. In this work, we evaluated the effects of PSN-1 on amastigotes of L. amazonensis inside macrophages infected in vitro. We assessed the production of hydrogen peroxide and nitric oxide, as well as the levels of inflammatory and immunomodulatory markers (TGF-ß, TNF-α and IL-12), in infected and non-infected macrophages treated with PSN-1. Treatment with PSN-1 decreased the number of infected cells and the number of ingested amastigotes per cell when compared with the untreated cells. At 32 µM (64 µg/mL), PSN-1 reduced hydrogen peroxide levels in both infected and uninfected macrophages, whereas it had little effect on NO production or TGF-ß release. The effect of PSN-1 on IL-12 and TNF-α secretion depended on its concentration, but, in general, their levels tended to increase as PSN-1 concentration increased. Further in vitro and in vivo studies are needed to clarify the mechanisms of action of PSN-1 and its interaction with the immune system aiming to develop pharmacological applications.

Mechanical Periodontal Therapy Recovered the Phagocytic Function of Monocytes in Periodontitis.

BACKGROUND: Several studies have focused on the association between periodontitis and systemic implications; however, the biological mechanisms of the immune responses before and after periodontal therapy involved in this relationship, such as phagocytic functions, remain unclear. OBJECTIVES: This study aimed to investigate whether periodontal treatment improves the phagocytic function of blood monocytes in patients with severe periodontitis. Materials and Methods. A nonrandomized sample of 55 participants was enrolled in the study. Two groups were studied: control (n = 27, healthy subjects without periodontal disease) and patients (n = 27, healthy subjects without periodontal disease) and patients (. RESULTS: Periodontitis induced impaired phagocytosis by monocytes. Phagocytosis at baseline was significantly lower in periodontitis patients [median, 13.2 (range of 7.1 to 20.8) and 60.7 (40.6 to 88.6)] than in controls [27.4 (15.5 to 40.5)] and 98 (68.2 to 122.9)] for nonsensitized or sensitized samples, respectively. After supportive therapy, patients showed a significant enhancement of phagocytic functions [33.7 (14.6 to 53.2) and 108.5 (99.6 to 159.5)] for nonsensitized and sensitized samples, respectively. Periodontal treatment increased the phagocytic capacity to a level similar to that observed in the control group and improved the capacity of phagocytes to produce superoxide anion. CONCLUSIONS: The results suggest that periodontal therapy in patients with severe periodontitis provides a state of homeostasis due to the reestablishment of phagocytic function and increased production of NBT (Regional Registry No. RBR-24T799; Universal Registry No. U1111-1133-5512).

Antiplasmodial and antileishmanial activities of phylloseptin-1, an antimicrobial peptide from the skin secretion of Phyllomedusa azurea (Amphibia).

The development of drug resistance by infectious agents represents a major hindrance for controlling parasitic diseases and has stimulated the search for new compounds. We have previously shown that phylloseptin-1 (PS-1), a cationic peptide from the skin secretion of Phyllomedusa azurea, exhibited potent antimicrobial activity. Now we evaluate the effect of PS-1 on Leishmania amazonensis and Plasmodium falciparum. Concentrations as low as 0.5 microg/mL of PS-1 exhibited antileishmanial activity comparable to that of antimoniate of N-metilglucamine, while the antiplasmodial effect of PS-1 was evident at the concentration of 16 microg/mL, and reached an activity comparable to that of artesunate, at the concentration of 64 microg/mL. The high antiparasitic activity of PS-1, together with the unrelatedness of its chemical structure to any present antimicrobial drug, which prevents the development of cross-resistance, together with its non-toxicity to mammalian cells make this peptide a promising candidate for the treatment of malaria and leishmaniasis.

Impact of polyphenols in phagocyte functions.

Polyphenols are a broad group of substances with potential health benefits found in plant species. Several of these compounds are capable of influencing the activation of intracellular signaling pathways, such as NF-kB, MAPK and JAK-STAT, responsible for the production of various inflammatory mediators such as tumor necrosis factor α (TNF-α) and interleukin 1 beta (IL-1ß) and 12 (IL-12), enzymes involved in the production of reactive species such as inducible nitric oxide synthase (iNOS) and superoxide dehydrogenase (SOD), as well as enzymes involved in the production of eicosanoids, such as cyclooxygenase (COX) and lipoxygenase (LO). There is increased interest in the use of polyphenol-rich foods because of their immunomodulatory effect; however, the mechanisms used during macrophage responses are extremely complex and little is known about the effects of polyphenols on these cells. As such, this review summarizes the current view of polyphenol influences on macrophages.

Atomic Force Microscopy Is a Potent Technique to Study Eosinophil Activation.

Eosinophils are multifunctional cells with several functions both in healthy individuals, and those with several diseases. Increased number and morphological changes in eosinophils have been correlated with the severity of an acute asthma exacerbation. We measured eosinophils obtained from healthy controls and individuals with acute asthma using atomic force microscopy (AFM). In the control samples, cells showed more rounded morphologies with some spreading, while activated cells from symptomatic individuals were spreading, and presenting emission of multiple pseudopods. Eosinophils presenting separate granules close to the cells suggesting some degranulation was also increased in asthma samples. In comparison to histopathological techniques based on brightfield microscopy, AFM showed considerably more details of these morphological changes, making the technique much more sensitive to detect eosinophil morphological changes that indicate functional alteration of this cell. AFM could be an important tool to evaluate diseases with alterations in eosinophil functions.

Anti-EpCAM antibodies for detection of metastatic carcinoma in effusions and peritoneal wash.

Epithelial cell adhesion molecule (EpCAM) has been used as diagnostic/prognostic marker and therapeutic target. The aim of the present study was to compare immunoreactivity of antibodies against distinct epitopes in the ectodomain of EpCAM for detection of carcinoma from different primary sites and of different histological types in effusions and peritoneal wash. Two antibodies against epitopes in the EGF-like domain I (clones Moc-31 and Ber-EP4) and one antibody against the epitope in the cysteine-poor region (158210) of EpCAM were used (all commercially available). Independently of the clone used, EpCAM overexpression was observed in almost all samples when all the adenocarcinoma samples were analyzed together. By using Moc-31, EpCAM overexpression was observed in all samples of adenocarcinoma. Absence of EpCAM overexpression was observed in a few adenocarcinoma samples at some sites of tumor origin, including ovary, breast and stomach, when Ber-EP4 and 158210 were used. Regarding carcinomas aside from adenocarcinomas, histological types, such as squamous cell, urothelial and small cell carcinoma showed different degrees of EpCAM expression according to the antibody used. In squamous cell carcinoma, overexpression was observed only with the clone 158210. It was concluded that, overall, most samples of metastatic carcinoma from effusions showed overexpression of EpCAM. However, there are significant variations in its detection according to the primary site, histological type of the carcinoma and depending on the antibody used. Thus, the use of more than one type of anti-EpCAM antibody would increase the chance of its detection in metastatic carcinoma effusion.