Polymorphisms and expression of inflammasome genes are associated with the development and severity of rheumatoid arthritis in Brazilian patients.

OBJECTIVE: In the present study, we analyzed the possible association of inflammasome gene variants and expression to rheumatoid arthritis (RA)'s development and severity in the Brazilian population. MATERIALS AND METHODS: Thirteen single nucleotide polymorphisms within six inflammasome genes (NLRP1, NLRP3, NLRC4, AIM2, CARD8, CASP1) as well as IL1B and IL18 genes in two different Brazilian populations (from Northeast and Southeast Brazil) were analyzed. We also evaluated inflammasome gene expression profile in resting and LPS + ATP-treated monocytes from RA patients and healthy individuals. For genetic association study, 218 patients and 307 healthy controls were genotyped. For gene expression study, inflammasome genes mRNA levels of 12 patients and ten healthy individuals were assessed by qPCR. RESULTS: Our results showed that rs10754558 NLRP3 and rs2043211 CARD8 polymorphisms are associated with RA development (p value = 0.044, OR = 1.77, statistical power = 0.999) and severity measured by Health Assessment Questionnaire (HAQ) (p value = 0.03), respectively. Gene expression analyses showed that RA patients display activation of CASP1, IL1B and IL1R genes independently of LPS + ATP activation. In LPS + ATP-treated monocytes, NLRP3 and NLRC4 expressions were also significantly higher in patients compared with controls. CONCLUSIONS: The first reported results in Brazilian populations support the role of inflammasome in the development of RA.

Smartphone based monitoring system for long-term sleep assessment.

The diagnosis of sleep disorders, highly prevalent in Western countries, typically involves sophisticated procedures and equipment that are highly intrusive to the patient. The high processing capabilities and storage capacity of current portable devices, together with a big range of available sensors, many of them with wireless capabilities, create new opportunities and change the paradigms in sleep studies. In this work, a smartphone based sleep monitoring system is presented along with the details of the hardware, software and algorithm implementation. The aim of this system is to provide a way for subjects, with no pre-diagnosed sleep disorders, to monitor their sleep habits, and on the initial screening of abnormal sleep patterns.

Hypnogram and sleep parameter computation from activity and cardiovascular data.

The automatic computation of the hypnogram and sleep Parameters, from the data acquired with portable sensors, is a challenging problem with important clinical applications. In this paper, the hypnogram, the sleep efficiency (SE), rapid eye movement (REM), and nonREM (NREM) sleep percentages are automatically estimated from physiological (ECG and respiration) and behavioral (Actigraphy) nocturnal data. Two methods are described; the first deals with the problem of the hypnogram estimation and the second is specifically designed to compute the sleep parameters, outperforming the traditional estimation approach based on the hypnogram. Using an extended set of features the first method achieves an accuracy of 72.8%, 77.4%, and 80.3% in the detection of wakefulness, REM, and NREM states, respectively, and the second an estimation error of 4.3%, 9.8%, and 5.4% for the SE, REM, and NREM percentages, respectively.

Sleep and wakefulness state detection in nocturnal actigraphy based on movement information.

Wrist actigraphy (ACT) is a low-cost and well-established technique for long-term monitoring of human activity. It has a special relevance in sleep studies, where its noninvasive nature makes it a valuable tool for behavioral characterization and for the detection and diagnosis of some sleep disorders. The traditional sleep/wakefulness state estimation algorithms from the nocturnal ACT data are unbalanced from a sensitivity and specificity points of view since they tend to overestimate sleep state, with severe consequences from a diagnosis point of view. They usually maximize the overall accuracy that does not take into account the highly unbalanced state distribution. In this paper, a method is proposed to appropriately deal with this unbalanced problem, achieving similar sensitivity and specificity scores in the state estimation process. The proposed method combines two linear discriminant classifiers, trained with two different criteria involving movement detection to generate a first state estimate. This result is then refined by a Hidden Markov Model-based algorithm. The global accuracy, the sensitivity, and the specificity of the method are 77.8%, 75.6%, and 81.6%, respectively, performing better than the tested algorithms. If the performance is assessed only for movement periods, this improvement is even higher.

Previous contact with Strongyloides venezuelensis contributed to prevent insulitis in MLD-STZ diabetes.

Epidemiological and experimental studies support the idea that helminth infections can induce a protective effect against the development of autoimmune and allergic diseases. In this study we characterized the immune response induced by Strongyloides venezuelensis infection in C57BL/6 mice and then evaluated the effect of a previous contact with this helminth in the outcome of type 1 diabetes. Animals were initially infected with 2000 L3 larvae from S. venezuelensis and euthanized 22 days later. An acute phase, identified by a high amount of eggs per gram of feces, was established between days 7 and 9 post-infection. Recovery from infection was associated with a Th2 polarized response characterized by a significant level of serum IgG1 specific antibodies and also a significant production of IL-5 and IL-10 by spleen cells stimulated with S. venezuelensis soluble antigen. Immunization with soluble S. venezuelensis antigen associated with complete Freund's adjuvant followed by infection with S. venezuelensis protected mice from diabetes development induced by streptozotocin. Protection was characterized by a higher body weight gain, lower glycemic levels, much less severe insulitis and preserved insulin production. Together, these results indicate that S. venezuelensis contributed to protect C57BL/6 mice against experimental diabetes induced by streptozotocin.

Developmental exposure to diuron causes splenotoxicity in male Sprague-Dawley rat pups.

This study investigated whether perinatal exposure to diuron [3-(3,4-dichlorophenyl)-1-1-dimethylurea] might exert adverse effects on rat lymphoid organs. Pregnant Sprague-Dawley (SD) rats were exposed to diuron at 500, 750 or 1250 ppm in the diet from gestational days (GD) 12-21 and during lactation. At postnatal day (PND) 42, male pups were euthanized and thymus, spleen, mesenteric lymph node and femur were collected for histopathological analysis. Food consumption and body weight gain were significantly reduced in dams exposed to 1250 ppm during gestation period. Also, Diuron at 750 and 1250 ppm produced: (1) increased relative spleen weight associated histologically with severe congestion in red pulp, (2) enhanced extramedullary hematopoiesis and hemosiderosis as well as (3) depletion of lymphoid follicles in white pulp. Flow cytometric analysis revealed a significant reduction in B lymphocytes (CD45RA+) in male pups but T lymphocytes (CD4+, CD8+ and CD4+/CD8+) were not markedly affected. Thus, data suggest that Diuron-induced maternal toxicity in dams exposed to high dose and perinatal exposure to this herbicide produced spleen toxicity as evidenced by a reduction in B lymphocyte number in male SD pups.

Prevention of experimental diabetes by Uncaria tomentosa extract: Th2 polarization, regulatory T cell preservation or both?

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria tomentosa (Willd.) DC (Rubiaceae) is a species native to the Amazon rainforest and surrounding tropical areas that is endowed with immunomodulatory properties and widely used around the world. In this study we investigated the immunomodulatory potential of Uncaria tomentosa (UT) aqueous-ethanol extract on the progression of immune-mediated diabetes. MATERIALS AND METHODS: C57BL/6 male mice were injected with MLDS (40 mg/kg) and orally treated with UT at 10-400mg/kg during 21 days. Control groups received MLDS alone or the respective dilution vehicle. Pancreatic mononuclear infiltrate and ß-cell insulin content were analyzed by HE and immunohistochemical staining, respectively, and measured by digital morphometry. Lymphocyte immunophenotyping and cytokine production were determined by flow cytometry analysis. RESULTS: Treating the animals with 50-400mg/kg of UT caused a significant reduction in the glycemic levels, as well as in the incidence of diabetes. The morphometric analysis of insulitis revealed a clear protective effect. Animals treated with UT at 400mg/kg presented a higher number of intact islets and a significant inhibition of destructive insulitis. Furthermore, a significant protection against the loss of insulin-secreting presented ß-cells was achieved, as observed by a careful immunohistochemical evaluation. The phenotypic analysis indicated that the groups treated with higher doses (100-400mg/kg) presented CD4(+) and CD8(+) T-cell values similar to those observed in healthy animals. These same higher doses also increased the number of CD4(+)CD25(+)Foxp3(+) regulatory T-cells. Moreover, the extract modulated the production of Th1 and Th2, with increased levels of IL-4 and IL-5. CONCLUSIONS: The extract was effective to prevent the progression of immune-mediated diabetes by distinct pathways.

Diuron exposure induces systemic and organ-specific toxicity following acute and sub-chronic exposure in male Wistar rats.

Diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] is a substitute urea herbicide widely used on agricultural crops with potential mutagenic, teratogenic, reproductive and carcinogenic effects. Nonetheless, its toxic potential on the immune system needs a detailed assessment. Thus, in order to evaluate the adverse effect of this herbicide on lymphohematopoietic organs and macrophage activity, male Wistar rats were orally treated with Diuron at 125, 1250 and 2500 ppm for 14, 28 or 90 days. General signs of toxicity were observed in Diuron-treated groups (1250 and 2500 ppm), including reduced food intake and body weight gain, as well as higher relative weights for spleen, kidneys and liver (28 and 90-day toxicity studies) and elevated serum levels of ALT, albumin, total protein, creatinine and urea (28-day toxicity study). Diuron exposure caused a severe depletion of splenic white pulp compartments and cellularity, followed by a decreased number of CD4(+) T lymphocytes, increased extramedullary hematopoiesis and deposition of hemosiderin in red pulp. Despite alteration in macrophage spreading, the macrophagic activity was not significantly affected by the herbicide. Under these experimental conditions, the results suggest that Diuron exerts systemic and target-organ toxicity, mainly at higher concentration.

Uncaria tomentosa aqueous-ethanol extract triggers an immunomodulation toward a Th2 cytokine profile.

Uncaria tomentosa (Willd.) DC (Rubiaceae) is a large woody vine that is native to the Amazon and Central American rainforests and is used widely in traditional medicine for its immunomodulatory and antiinflammatory activities. The present work used in vivo immunotoxic and in vitro immunomodulatory experiments to investigate the effects of a pentacyclic oxindole alkaloid extract from U. tomentosa bark on lymphocyte phenotype, Th1/Th2 cytokine production, cellular proliferation and cytotoxicity. For the in vivo immunotoxicity testing, BALB/c male mice were treated once a day with 125, 500 or 1250 mg/kg of U. tomentosa extract for 28 days. For the in vitro protocol, lymphocytes were cultured with 10-500 µg/mg of the extract for 48 h. The extract increased the cellularity of splenic white pulp and the thymic medulla and increased the number of T helper lymphocytes and B lymphocytes. Also, a large stimulatory effect on lymphocyte viability was observed. However, mitogen-induced T lymphocyte proliferation was significantly inhibited at higher concentrations of U. tomentosa extract. Furthermore, an immunological polarization toward a Th2 cytokine profile was observed. These results suggest that the U. tomentosa aqueous-ethanol extract was not immunotoxic to mice and was able to modulate distinct patterns of the immune system in a dose-dependent manner.

Statistical characterization of actigraphy data during sleep and wakefulness states.

Human activity can be measured with actimetry sensors used by the subjects in several locations such as the wrists or legs. Actigraphy data is used in different contexts such as sports training or tele-medicine monitoring. In the diagnosis of sleep disorders, the actimetry sensor, which is basically a 3D axis accelerometer, is used by the patient in the non dominant wrist typically during an entire week. In this paper the actigraphy data is described by a weighted mixture of two distributions where the weight evolves along the day according to the patient circadian cycle. Thus, one of the distributions is mainly associated with the wakefulness state while the other is associated with the sleep state. Actigraphy data, acquired from 20 healthy patients and manually segmented by trained technicians, is used to characterize the acceleration magnitude during sleep and wakefulness states. Several mixture combinations are tested and statistically validated with conformity measures. It is shown that both distributions can co-exist at a certain time with varying importance along the circadian cycle.

Automatic annotation of actigraphy data for sleep disorders diagnosis purposes.

The diagnosis of Sleep disorders, highly prevalent in the western countries, typically involves sophisticated procedures and equipments that are intrusive to the patient. Wrist actigraphy, on the contrary, is a non-invasive and low cost solution to gather data which can provide valuable information in the diagnosis of these disorders. The acquired data may be used to infer the Sleep/Wakefulness (SW) state of the patient during the circadian cycle and detect abnormal behavioral patterns associated with these disorders. In this paper a classifier based on Autoregressive (AR) model coefficients, among other features, is proposed to estimate the SW state. The real data, acquired from 23 healthy subjects during fourteen days each, was segmented by expert medical personal with the help of complementary information such as light intensity and Sleep e-Diary information. Monte Carlo tests with a Leave-One-Out Cross Validation (LOOCV) strategy were used to assess the performance of the classifier which achieves an accuracy of 96%.

Optimization strategies for metabolic networks.

BACKGROUND: The increasing availability of models and data for metabolic networks poses new challenges in what concerns optimization for biological systems. Due to the high level of complexity and uncertainty associated to these networks the suggested models often lack detail and liability, required to determine the proper optimization strategies. A possible approach to overcome this limitation is the combination of both kinetic and stoichiometric models. In this paper three control optimization methods, with different levels of complexity and assuming various degrees of process information, are presented and their results compared using a prototype network. RESULTS: The results obtained show that Bi-Level optimization lead to a good approximation of the optimum attainable with the full information on the original network. Furthermore, using Pontryagin's Maximum Principle it is shown that the optimal control for the network in question, can only assume values on the extremes of the interval of its possible values. CONCLUSIONS: It is shown that, for a class of networks in which the product that favors cell growth competes with the desired product yield, the optimal control that explores this trade-off assumes only extreme values. The proposed Bi-Level optimization led to a good approximation of the original network, allowing to overcome the limitation on the available information, often present in metabolic network models. Although the prototype network considered, it is stressed that the results obtained concern methods, and provide guidelines that are valid in a wider context.

Decreased production of TNF-alpha by lymph node cells indicates experimental autoimmune encephalomyelitis remission in Lewis rats.

Experimental autoimmune encephalomyelitis (EAE) is mediated by CD4+ Th1 cells that mainly secrete IFN-gamma and TNF-alpha, important cytokines in the pathophysiology of the disease. Spontaneous remission is, in part, attributed to the down regulation of IFN-gamma and TNF-alpha by TGF-beta. In the current paper, we compared weight, histopathology and immunological parameters during the acute and recovery phases of EAE to establish the best biomarker for clinical remission. Female Lewis rats were immunised with myelin basic protein (MBP) emulsified with complete Freund's adjuvant. Animals were evaluated daily for clinical score and weight prior to euthanisation. All immunised animals developed the expected characteristics of EAE during the acute phase, including significant weight loss and high clinical scores. Disease remission was associated with a significant reduction in clinical scores, although immunised rats did not regain their initial weight values. Brain inflammatory infiltrates were higher during the acute phase. During the remission phase, anti-myelin antibody levels increased, whereas TNF-alpha and IFN-gamma production by lymph node cells cultured with MBP or concanavalin A, respectively, decreased. The most significant difference observed between the acute and recovery phases was in the induction of TNF-alpha levels in MBP-stimulated cultures. Therefore, the in vitro production of this cytokine could be used as a biomarker for EAE remission.

Decreased production of TNF-alpha by lymph node cells indicates experimental autoimmune encephalomyelitis remission in Lewis rats

Experimental autoimmune encephalomyelitis (EAE) is mediated by CD4+ Th1 cells that mainly secrete IFN-γ and TNF-α, important cytokines in the pathophysiology of the disease. Spontaneous remission is, in part, attributed to the down regulation of IFN-γ and TNF-α by TGF-β. In the current paper, we compared weight, histopathology and immunological parameters during the acute and recovery phases of EAE to establish the best biomarker for clinical remission. Female Lewis rats were immunised with myelin basic protein (MBP) emulsified with complete Freund's adjuvant. Animals were evaluated daily for clinical score and weight prior to euthanisation. All immunised animals developed the expected characteristics of EAE during the acute phase, including significant weight loss and high clinical scores. Disease remission was associated with a significant reduction in clinical scores, although immunised rats did not regain their initial weight values. Brain inflammatory infiltrates were higher during the acute phase. During the remission phase, anti-myelin antibody levels increased, whereas TNF-α and IFN-γ production by lymph node cells cultured with MBP or concanavalin A, respectively, decreased. The most significant difference observed between the acute and recovery phases was in the induction of TNF-α levels in MBP-stimulated cultures. Therefore, the in vitro production of this cytokine could be used as a biomarker for EAE remission.