Copaiba oil minimizes inflammation and promotes parenchyma re-epithelization in acute allergic asthma model induced by ovalbumin in BALB/c mice.

Introduction: Asthma is a condition of airflow limitation, common throughout the world, with high mortality rates, especially as it still faces some obstacles in its management. As it constitutes a public health challenge, this study aimed to investigate the effect of copaiba oil (e.g., Copaifera langsdorffii), as a treatment resource, at doses of 50 and 100 mg/kg on certain mediators of acute lung inflammation (IL-33, GATA3, FOXP3, STAT3, and TBET) and early mechanisms of lung remodeling (degradation of elastic fiber tissues, collagen deposition, and goblet cell hyperplasia). Methods: Using an ovalbumin-induced acute allergic asthma model in BALB/c mice, we analyzed the inflammatory mediators through immunohistochemistry and the mechanisms of lung remodeling through histopathology, employing orcein, Masson's trichrome, and periodic acid-Schiff staining. Results: Copaiba oil treatment (CO) reduced IL-33 and increased FOXP3 by stimulating the FOXP3/GATA3 and FOXP3/STAT3 pathways. Additionally, it upregulated TBET, suggesting an additional role in controlling GATA3 activity. In the respiratory epithelium, CO decreased the fragmentation of elastic fibers while increasing the deposition of collagen fibers, favoring epithelial restructuring. Simultaneously, CO reduced goblet cell hyperplasia. Discussion: Although additional research is warranted, the demonstrated anti-inflammatory and re-epithelializing action makes CO a viable option in exploring new treatments for acute allergic asthma.

Kaurenoic acid nanocarriers regulates cytokine production and inhibit breast cancer cell migration.

Breast cancer is the type of cancer with the highest incidence in women around the world. Noteworthy, the triple-negative subtype affects 20% of the patients while presenting the highest death rate among subtypes. This is due to its aggressive phenotype and the capability of invading other tissues. In general, tumor-associated macrophages (TAM) and other immune cells, are responsible for maintaining a favorable tumor microenvironment for inflammation and metastasis by secreting several mediators such as pro-inflammatory cytokines IL-1ß, IL-6, and TNF-α, chemokines like CCL2, and other proteins, as metalloproteinases of matrix (MMP). On the other hand, immunomodulatory agents can interfere in the immune response of TAM and change the disease prognosis. In this work, we prepared nanostructured lipid carriers containing kaurenoic acid (NLC-KA) to evaluate the effect on cytokine production in vitro of bone marrow-derived macrophages (BMDM) and the migratory process of 4 T1 breast cancer cells. NLC-KA prepared from a blend of natural lipids was shown to have approximately 90 nm in diameter with low polydispersity index. To test the effect on cytokine production in vitro in NLC-KA treated BMDM, ELISA assay was performed and pro-inflammatory cytokines IL-1ß, IL-6, and TNF-α were quantified. The formulation reduced the secretion of IL-1ß and TNF-α cytokines while presenting no hemolytic activity. Noteworthy, an anti-migratory effect in 4 T1 breast cancer cells treated with NLC-KA was observed in scratch assays. Further, MMP9 and CCL2 gene expressions in both BMDM and 4 T1 treated cells confirmed that the mechanism of inhibition of migration is related to the blockade of this pathway by KA. Finally, cell invasion assays confirmed that NLC-KA treatment resulted in less invasiveness of 4 T1 cells than control, and it is independent of CCL2 stimulus or BMDM direct stimulus. Ultimately, NLC-KA was able to regulate the cytokine production in vitro and reduce the migration of 4 T1 breast cancer cells by decreasing MMP9 gene expression.

Cross-sectional assessment of mild cognitive impairment in pre-dialysis chronic kidney disease and its association with inflammation and changes seen on MRI: what the eyes cannot see / Avaliação transversal do comprometimento cognitivo leve em doença renal crônica pré-dialítica e associação com inflamação e alterações na ressonância magnética: o que os olhos não veem

Abstract Introduction: Mild cognitive impairment (MCI) is a prevalent and underdiagnosed condition in chronic kidney disease (CKD), that shares common pathophysiological factors such as chronic inflammation. Objective: To evaluate the association of MCI in CKD stages 1-5 using inflammatory markers and changes by magnetic resonance imaging (MRI). Patients and Methods: Cross-sectional study in adult patients with pre-dialysis CKD. MCI was assessed by the Montreal Cognitive Assessment (MoCA) and the estimated glomerular filtration rate (eGFR) by the Chronic Kidney Disease Epidemiology Collaboration equation. Sociodemographic and clinical data were collected from medical records. The cytokines IL-4, IL-6, IL-17, TNF-α and hs-CRP were determined. Brain MRI was performed in a 1.5 Tesla device, without paramagnetic contrast. A descriptive analysis followed by a comparison of abnormal versus normal MoCA scores among all studied variables. A linear regression analysis was performed using MoCA as a dependent variable, adjusted for confounding factors. Results: Of 111 invited patients, eighty completed the neuropsychological assessment and 56 underwent MRI, and were included in the study. Mean age was 56.3 ± 8.3 years and 51.8% (n = 29) had altered MoCA. When compared to the group with normal MoCA, the group with altered MoCA had higher levels of IL-6 and IL-17. There was no correlation between altered MoCA with eGFR or with MRI abnormalities. Conclusão: MCI assessed by MoCA was prevalent in patients with pre-dialysis CKD, it was associated with inflammation and showed no correlation with MRI changes.

Resumo Introdução: O comprometimento cognitivo leve (CCL) é prevalente e subdiagnosticado na doença renal crônica (DRC), condição com a qual compartilha fatores fisiopatológicos como a inflamação crônica. Objetivo: Avaliar a associação do CCL na DRC estágios 1 a 5, com marcadores inflamatórios e alterações de exames de imagem por ressonância magnética (RM). Pacientes e métodos: Estudo transversal em pacientes adultos, com DRC pré-dialítica. CCL foi avaliado pelo Montreal Cognitive Assessment (MoCA) e a taxa de filtração glomerular estimada (TFGe), pela equação do CKD-EPI. Dados sociodemográficos e clínicos foram coletados nos prontuários médicos. Dosadas citocinas IL-4, IL-6, IL-17, o TNF-α e PCR-us. A RM do encéfalo foi realizada em aparelho de 1,5 Tesla, sem contraste. Realizada análise descritiva seguida por comparação de pontuações do MoCA anormais versus normais entre todas as variáveis estudadas. A regressão linear foi realizada usando MoCA como uma variável dependente, ajustada para fatores de confusão. Resultados: De 111 pacientes convidados, oitenta completaram a avaliação neuropsicológica, 56 realizaram RM, tendo sido incluídos no estudo. A média de idade foi de 56,3 ± 8,3 anos e 51,8% (n = 29) apresentavam MoCA alterado. Quando comparado ao grupo MoCA normal, o grupo MoCA alterado apresentou níveis mais elevados de IL-6 e IL-17. Não houve correlação entre MoCA alterado com TFGe nem com anormalidades na RM. Nos modelos ajustados, a IL-6 foi preditor independente do MoCA alterado Conclusão: O CCL avaliado pelo MoCA foi prevalente em pacientes com DRC pré-dialítica, associou-se com inflamação e não apresentou correlação com alterações da RM.

Impact of an educational intervention in the management of individuals with uncontrolled type 2 diabetes mellitus using insulin therapy.

OBJECTIVE: To investigate the effects of problematizing intervention in the treatment of individuals with type 2 diabetes mellitus. METHODOLOGY: A randomized clinical trial was conducted in 41 patients ages 18 to 64 with type 2 diabetes who were treated with insulin and had glycosylated hemoglobin greater than 7.0%. The mean age of participants was 55.9 (SD = 5.49). A high percentage of patients had comorbidities such as hypertension (92.7%), dyslipidemia (68.3%), overweight (95%), retinopathy (41%), and neuropathy (39%). The patients in the intervention group participated in 6 educational groups using problematization methodology, whereas the patients in the control group attended only routine consultations. Sociodemographic, clinical, behavioral, and lifestyle variables were assessed. RESULTS: After 6 months of follow-up, no statistically significant difference in glycemic control and anthropometric parameters was observed between participants in either study group. The intervention group showed an increase in knowledge about the disease, and an improvement in total cholesterol and uric acid levels. CONCLUSION: The use of a problematizing intervention provided an improvement in behavioral as well as specific clinical parameters, compared to routine diabetes care. However, longer follow-up time for these patients could bring benefits regarding glycemic control.

Cross-sectional assessment of mild cognitive impairment in pre-dialysis chronic kidney disease and its association with inflammation and changes seen on MRI: what the eyes cannot see.

INTRODUCTION: Mild cognitive impairment (MCI) is a prevalent and underdiagnosed condition in chronic kidney disease (CKD), that shares common pathophysiological factors such as chronic inflammation. OBJECTIVE: To evaluate the association of MCI in CKD stages 1-5 using inflammatory markers and changes by magnetic resonance imaging (MRI). PATIENTS AND METHODS: Cross-sectional study in adult patients with pre-dialysis CKD. MCI was assessed by the Montreal Cognitive Assessment (MoCA) and the estimated glomerular filtration rate (eGFR) by the Chronic Kidney Disease Epidemiology Collaboration equation. Sociodemographic and clinical data were collected from medical records. The cytokines IL-4, IL-6, IL-17, TNF-α and hs-CRP were determined. Brain MRI was performed in a 1.5 Tesla device, without paramagnetic contrast. A descriptive analysis followed by a comparison of abnormal versus normal MoCA scores among all studied variables. A linear regression analysis was performed using MoCA as a dependent variable, adjusted for confounding factors. RESULTS: Of 111 invited patients, eighty completed the neuropsychological assessment and 56 underwent MRI, and were included in the study. Mean age was 56.3 ± 8.3 years and 51.8% (n = 29) had altered MoCA. When compared to the group with normal MoCA, the group with altered MoCA had higher levels of IL-6 and IL-17. There was no correlation between altered MoCA with eGFR or with MRI abnormalities. CONCLUSÃO: MCI assessed by MoCA was prevalent in patients with pre-dialysis CKD, it was associated with inflammation and showed no correlation with MRI changes.

Seroprevalence estimate and risk factors for Coxiella burnetii infections among humans in a highly urbanised Brazilian state.

BACKGROUND: Q fever is among the top 13 global priority zoonoses, however, it is still neglected and under-reported in most of the world, including Brazil. Thus, we evaluated the seroprevalence of and the risk factors for Coxiella burnetii infections in humans from Minas Gerais, a highly urbanised Brazilian state. METHODS: Coxiella burnetii was searched for patient samples (n=437), which were suspected of then later confirmed as negative for dengue fever, by the indirect immunofluorescence technique and real-time PCR. Risk factors for infections and spatial clusters for both C. burnetii-seropositive individuals and livestock concentration were evaluated. RESULTS: We found that 21 samples (4.8%; 95% CI 3.0 to 7.2%) were reactive for at least one class of anti-C. burnetii antibodies (titer of ≥64), with rural residence (p=0.036) being a risk factor. Also, two spatial clusters of seropositivity were found within a significant area by Scan, and a probable relationship between the Scan result and the livestock concentration by area was found. CONCLUSIONS: Seropositive individuals were associated with rural residence, with a likely relationship with the livestock concentration. Thus, this study establishes baseline figures for C. burnetii seroprevalence in humans in a state of Brazil, allowing the monitoring of trends and setting of control targets, as well as more representative longitudinal and risk analysis studies.

Adjusting RT-qPCR conditions to avoid unspecific amplification in SARS-CoV-2 diagnosis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and quickly spread around the world, forcing global health authorities to develop protocols for its diagnosis. Here we report dimer formation in the N2 primers-probe set (CDC 2019-nCoV Real-Time RT-PCR) used in the diagnostic routine, and propose alternatives to reduce dimerization events. Late unspecific amplifications were visualized in 56.4% of negative samples and 57.1% of no-template control, but not in positive samples or positive control. In silico analysis and gel electrophoresis confirmed the dimer formation. The RT-qPCR parameters were optimized and the late unspecific amplifications decreased to 11.5% in negative samples and no-template control. The adjustment of PCR parameters was essential to reduce the risk of false-positives results and to avoid inclusive results requiring repeat testing, which increases the costs and generates delays in results or even unnecessary requests for new samples.

Infliximab-induced remission improves physical activity in patients with active Crohn's disease.

AIM: To compare the level of physical activity (PA), exercise capacity, and body composition before and after infliximab-induced clinical remission in patients with Crohn's disease (CD). METHODS: This prospective longitudinal study evaluated 44 adult outpatients with active CD before infliximab administration and 24 weeks after infliximab therapy. The patients were evaluated for PA in daily life, exercise capacity, muscle strength, and body composition. RESULTS: 38 (86.4%) patients achieved infliximab-induced remission at 24 weeks and presented an increment in the number of steps taken of 1092 (7440±2980 vs. 6348±3177, respectively; p=0.006). The inactive time was reduced when compared to the baseline value (454.2±106.3 vs. 427.9±97.8, respectively; p=0.033). There was no difference in the distance walked before and after infliximab therapy, while there was an increase in the fat mass index in responders to infliximab compared to the baseline (19.1±7.6 vs. 14.9±5.8; p=0.001). CONCLUSIONS: Infliximab-induced remission was shown to be effective for increasing physical activity by improving the number of steps and reducing inactive time. The maintenance of clinical remission associated with incentives to regular PA may contribute to making these patients reach an ideal level of PA.

Infliximab-induced remission improves physical activity in patients with active Crohn's disease

SUMMARY AIM: To compare the level of physical activity (PA), exercise capacity, and body composition before and after infliximab-induced clinical remission in patients with Crohn's disease (CD). METHODS: This prospective longitudinal study evaluated 44 adult outpatients with active CD before infliximab administration and 24 weeks after infliximab therapy. The patients were evaluated for PA in daily life, exercise capacity, muscle strength, and body composition. RESULTS: 38 (86.4%) patients achieved infliximab-induced remission at 24 weeks and presented an increment in the number of steps taken of 1092 (7440±2980 vs. 6348±3177, respectively; p=0.006). The inactive time was reduced when compared to the baseline value (454.2±106.3 vs. 427.9±97.8, respectively; p=0.033). There was no difference in the distance walked before and after infliximab therapy, while there was an increase in the fat mass index in responders to infliximab compared to the baseline (19.1±7.6 vs. 14.9±5.8; p=0.001). CONCLUSIONS: Infliximab-induced remission was shown to be effective for increasing physical activity by improving the number of steps and reducing inactive time. The maintenance of clinical remission associated with incentives to regular PA may contribute to making these patients reach an ideal level of PA.

RESUMO OBJETIVO: Comparar o nível de atividade física (AF), capacidade de exercício e composição corporal antes e após remissão clínica induzida por infliximabe em pacientes com doença de Crohn (DC). MÉTODOS: Neste estudo longitudinal prospectivo, foram envolvidos 44 pacientes ambulatoriais adultos com DC ativa avaliados antes e depois de 24 semanas de terapia com infliximabe. Os pacientes foram avaliados quanto à AF, capacidade de exercício, força muscular e composição corporal. RESULTADOS: 38(86,4%) pacientes alcançaram remissão induzida por infliximabe em 24 semanas e apresentaram aumento no número de passos de 1092 (7440±2980 vs. 6348±3177, respectivamente; p=0,006). O tempo de inatividade foi reduzido quando comparado ao basal (454,2±106,3 vs. 427,9±97,8, respectivamente; p=0,033). Não houve diferença na distância percorrida antes e após a terapia com infliximabe, enquanto houve aumento no índice de massa gorda nos respondedores ao infliximabe em comparação ao basal (19,1±7,6 vs. 14,9±5,8; p=0,001). CONCLUSÕES: A remissão induzida pelo infliximabe mostrou-se eficaz no aumento da atividade física, melhorando o número de passos e reduzindo o tempo inativo. A manutenção da remissão clínica associada a incentivos à AF regular pode contribuir para que esses pacientes atinjam um nível ideal de AF.

DPP4 activity is related to body weight and central fat in postmenopausal women.

AIMS: Dipeptidyl peptidase-4 (DPP4) is a new adipokine increased in central obesity and related to insulin resistance (IR). Postmenopausal (PM) state may be associated with increase in body weight and central fat distribution. We hypothesize that DPP4 is increased in PM women. MATERIALS AND METHODS: Twenty-two non-obese PM and 22 non-obese premenopausal women (PreM), were evaluated. DPP4 activity, lipid profile, HbA1c, FSH, estradiol and sex hormone-binding globulin (SHBG) were measured; an oral glucose tolerance test (OGTT) was performed and IR calculated. Body composition was assessed by dual X-ray absorptiometry (DXA). Correlations between DPP4 and the anthropometric and metabolic variables and body fat distribution were studied. RESULTS: DPP4 activity was not different between the two groups (PM 5309 ±â€¯650 vs PreM 5387 ±â€¯704 RLU; p = 0,70). In the PM group there was a significant correlation between DPP4 and body weight (r = 0,498; p = 0,03; n = 22) and trunk fat (r = 0,477; p = 0,03; n = 21). There was also a trend for correlation with android (r = 0,418; p = 0,06; n = 21) and total fat (r = 0,409; p = 0,06; n = 21). When stratified by BMI, DPP4 was significantly higher in PM women with BMI ≥25 kg/m2 (p = 0,02). CONCLUSION: DPP4 was not increased in PM but is associated with body weight and body fat centralization.

Comparative analysis for strength serum sodium and potassium in three different methods: Flame photometry, ion-selective electrode (ISE) and colorimetric enzymatic.

BACKGROUND: Hydroelectrolytic disorders are common in clinical situations and may be harmful to the patient, especially those involving plasma sodium and potassium dosages. Among the possible methods for the dosages are flame photometry, ion-selective electrode (ISE) and colorimetric enzymatic method. METHODS: We analyzed 175 samples in the three different methods cited from patients attending the laboratory of the University Hospital of the Federal University of Juiz de Fora. The values obtained were statistically treated using SPSS 19.0 software. The present study aims to evaluate the impact of the use of these different methods in the determination of plasma sodium and potassium. RESULTS: The averages obtained for sodium and potassium measurements by flame photometry were similar (P > .05) to the means obtained for the two electrolytes by ISE. The averages obtained by the colorimetric enzymatic method presented statistical difference in relation to ISE, both for sodium and potassium. In the correlation analysis, both flame photometry and colorimetric enzymatic showed a strong correlation with the ISE method for both dosages. CONCLUSION: At the first time in the same work sodium and potassium were analyzed by three different methods and the results allowed us to conclude that the methods showed a positive and strong correlation, and can be applied in the clinical routine.

Is DPP4 activity increased in PCOS?

AIMS: Dipeptidyl peptidase-4 (DPP4) is an adipokine with greater expression in visceral fat and related with insulin resistance (IR). Polycystic ovary syndrome (PCOS) is also associated with IR. Our study aims to evaluate DPP4 activity in PCOS. MATERIALS AND METHODS: Thirty PCOS patients were compared to 28 healthy women. Body composition by dual X-ray absorptiometry (DXA), plasma activity of DPP4 and biochemical variables were performed. All participants underwent an oral glucose tolerance test for insulin and glucose analysis. RESULTS: DPP4 activity was similar in both groups (PCOS 5823 ±â€¯926 vs Control 5501.8 ±â€¯975; p = 0.20). PCOS patients were more IR with lower levels of SHBG (32 vs 47, p = 0.02) and Matsuda index (15.6 vs 20.4, p = 0.03) and higher HOMA-IR (2.8 vs 1.7, p < 0.01), in addition to increased levels of testosterone (55 vs 25, p < 0.01). DPP4 was correlated to HbA1c (r = 0.279, p = 0.03), HDL-c (r = -0.28, p = 0.03) and SHBG (r = -0.256, p = 0.05). CONCLUSIONS: Although PCOS was well characterized as IR and hyperandrogenic, DPP4 was not different in this group. However, a relationship between DPP4 and markers of IR were found. More studies are warranted.

ß-Caryophyllene ameliorates the development of experimental autoimmune encephalomyelitis in C57BL/6 mice.

Multiple sclerosis is the most common autoimmune inflammatory and demyelinating disease of the central nervous system. The experimental autoimmune encephalomyelitis (EAE) is an appropriate and a well-establish model for studying the pathogenesis of MS. ß-caryophyllene (BCP), a natural sesquiterpene found in many plant species, is a potent anti-inflammatory compound. Herein we investigated the in vitro and in vivo immunomodulatory effects of BCP on C57BL/6 mice induced with EAE. BCP was in vitro evaluated (4, 20, and 40µM) on splenocytes obtained from EAE-induced C57BL/6 mice, and in vivo (25 or 50mg/kg/day) orally administered on EAE-mice. The clinical course, body weight, cytokines and oxygen radicals production were investigated in C57BL/6 EAE-mice. In vitro and in vivo immunological responses were evaluated by ELISA, and CNS sections were stained by hematoxylin and eosin methods The in vitro production of H2O2, NO, IFN-γ, and TNF- α was inhibited by BCP (20 and 40µM) in cultured cells from EAE-mice. BCP (25 and 50mg/kg/day) reduced clinical score and severity of EAE and inhibited H2O2, NO, TNF-α, IFN-γ and, IL-17 production. EAE-mice, orally treated with BCP (mainly at 50mg/kg/day), displayed levels of cytokines and clinical signs similar to animals with no EAE disease, demonstrating the therapeutic action of BCP on EAE animals. Histopathological and histomorphometric analysis confirmed that BCP treatment significantly reduced the numbers of inflammatory infiltrates and attenuated neurological damages in the CNS of EAE-mice.

Sarcopenia in patients with chronic kidney disease not yet on dialysis: Analysis of the prevalence and associated factors.

INTRODUCTION: Sarcopenia is a chronic condition that is associated with aging and characterized by a reduction of muscle mass, strength, and function. Sarcopenia is prevalent in patients with chronic kidney disease (CKD) and associated with increased morbidity and mortality, as well as cardiovascular complications. OBJECTIVES: To investigate the prevalence of sarcopenia in patients with CKD not yet on dialysis and its correlation with clinical and laboratory variables and inflammatory markers. METHODS: A total of 100 patients of both sexes aged over 18 were evaluated. Sarcopenia was defined using the criteria of the European Working Group on Sarcopenia in Older People (EWGSOP) and of the Foundation for the National Institutes of Health (FNIH) Sarcopenia Project. Sociodemographic and clinical data, activities of daily living, functional capacity, and physical activity were also evaluated. Inflammation was assessed by the serum levels of high-sensitivity C-reactive protein (hsCRP) and interleukin (IL) 4 and 6. RESULTS: The prevalence of sarcopenia was 11.9% and 28.7% using the EWGSOP and FNIH criteria, respectively. Sarcopenia was more prevalent in the more advanced stages of CKD (34.5% in stages 2 and 3A; and 65.5% in stages 3B, 4, and 5) and associated with worse performance in activities of daily living (p = 0.049), lower walking speeds (p < 0.001), and higher body mass indexes (BMIs) (p = 0.001) in the non-adjusted model. In addition, patients with sarcopenia had lower functional capacity (p = 0.012) and higher prevalence of physical inactivity (p = 0.041) compared with patients without sarcopenia. After adjustment for confounding variables, sarcopenia was still significantly correlated with walking speed (p = 0.004) and BMI (p = 0.002). HsCRP levels were inversely correlated with appendicular lean mass adjusted for BMI (p = 0.007) and were also positively associated with BMI (p = 0.001). IL4 levels were positively correlated with walking speed (p = 0.007) and lean mass in the lower limbs (p = 0.022). CONCLUSIONS: Sarcopenia is common in patients with CKD, particularly in the most advanced stages of the disease. We observed an association between the levels of inflammatory markers and peripheral lean body mass, physical performance, and BMI. This association between sarcopenia and modifiable factors highlights the importance of early diagnosis and the implementation of therapeutic measures to minimize adverse outcomes in patients with CKD not yet on dialysis.

Efeito da talidomida sobre o processo inflamatório e desmielinização na encefalomielite auto-imune experimental: estudo histopatológico e expressão de TNF-α e iNOS / Effect of thalidomide on the inflammatory process and demyelination in experimental autoimmune encephalomyelitis: histopathologic study and expression of TNF-α e iN

A encefalomielite autoimune experimental (EAE) é uma doença inflamatória e desmielinizante do sistema nervoso central (SNC) caracterizada por incapacidades temporárias ou permanentes. A patogênese envolve a reação auto-imune associada com a produção de citocinas pró inflamatórias, tais como o fator de necrose tumoral alfa (TNF-α). Esta citocina está associada com o aumento de radicais livres de oxigênio, como o óxido nítrico, liberados pelas células imunes ativadas. Além de aumentar a inflamação, tanto o fator de necrose tumoral, como o óxido nítrico causam lesão tecidual direta. Este estudo avaliou o efeito da talidomida na progressão clínica da doença, desenvolvimento da reação inflamatória e desmielinização. A expressão tecidual "in situ" do TNF-α e iNOS, uma enzima associada com a produção de óxido nítrico, foi investigada em amostras do SNC obtidos durante o desenvolvimento do modelo de EAE em ratos Lewis. Métodos: Ratos Lewis(n = 30) foram divididos em grupo de controle saudável (I), grupo experimental de encefalomielite autoimune (II) e o grupo tratado com talidomida (III). Os ratos foram monitorizados durante 15 dias para determinação da condição clínica, após este período, os animais foram eutanasiados e as amostras do sistema nervoso central foram obtidas para a realização de estudo histopatológico e imuno-histoquímico Resultados: Todos os animais do grupo II tiveram sintomas relacionados a EAE, enquanto apenas um do grupo tratado talidomida apresentaram alterações clínicas. O estudo histopatológico revelou que as amostras de SNC do grupo II apresentaram áreas de intenso infiltrado inflamatório mononuclear difuso e presença de áreas de desmielinização. No entanto, os animais tratados com talidomida apresentaram ocasionalmente um leve infiltrado inflamatório e bainhas de mielina bem organizadas. Além disso, a expressão de TNF-α e iNOS foram significativamente maiores no grupo II, quando comparado com o grupo tratado com a talidomida. Conclusões: Os resultados considerados em conjunto sustentam a hipótese de que a talidomida inibe a intensidade do processo inflamatório e desmielinização, assim como reduz a produção de mediadores inflamatórios modulando o desenvolvimento da encefalomielite auto-imune experimental em ratos Lewis.

Experimental autoimmune encephalomyelitis is a inflammatory and demyelinating disease of central nervous system (CNS) characterized by permanents or temporary disabilities. Its pathogenesis involves autoimmune reaction associated with the production of pro inflammatory cytokines such as tumor necrosis factor alpha (TNF-α). This cytokine is associated with increase of reactive oxygen free radicals, such as nitric oxide, released by activated immune cells. Besides enhancing inflammation, both tumor necrosis factor as nitric oxide cause pathologically direct destruction of proteins and enzyme oxidation. This study focuses on clinical disease progression, development of the inflammatory reaction and evaluation axonal myelination . The " in situ" tissue expression of the TNF-α and inducible nitric oxide synthase iNOS ,an enzyme associated with the production of nitric oxide , were also investigated in CNS samples obtained during the development of experimental autoimmune encephalomyelitis model in Lewis rats. Methods: Lewis rats were used to perform the classical model of EAE. The rats ( n=30) were divided into the healthy control group (I), experimental autoimmune encephalomyelitis group (II) and thalidomide treated group (III). The rats were monitored for 15 days for determination of clinical score , after this period , the animals were euthanized and samples were obtained from the central nervous system in which histopatological study and immunohistochemistry for SNC in situ detection of TNF-α and inducible nitric oxide synthase (iNOS) were performed. Results: All animals of group II had symptoms related to experimental encephalomyelitis , while only one of the thalidomide treated group showed clinical changes. The histopatological study revealead that SNC samples of group II presented areas of intense focal and diffuse mononuclear inflammation and the myelin sheaths were scarce and poorly stained. However, thalidomide treated rats presented occasionally a mild perivascular inflammatory infiltrate and myelin sheaths were organized and well evidenced. In addition, the expression of TNF-α and iNOS were significantly higher in the group II when compared with thalidomide treated group. Conclusions: The results taken together support the hypothesis that thalidomide inhibits the intensity of the inflammation and demyelination process and as well as reduces the production of inflammatory mediators influencing the development of experimental autoimmune encephalomyelitis in Lewis rats

Evaluation of platelet aggregation in the presence of antiphospholipid antibodies: anti-ß2GP1 and anticardiolipin.

INTRODUCTION: The antiphospholipid syndrome (APS) is an autoimmune condition characterized by recurrent arterial and venous thrombosis, besides obstetric complications. The pathogenesis is associated with the presence of antiphospholipid and/or anti-b2-glicoprotein I (anti-b2GPI) antibodies that appear to change the anticoagulant activity of b2GPI. Antibody-induced dimerization of b2GPI seems to be related to the induction of platelet aggregation, contributing to the development of thrombosis in APS. OBJECTIVES: The objective of the present study is to demonstrate the influence of antiphospholipid antibodies in platelet aggregation tests with different agonists (ADP, collagen, and adrenaline). METHODS: We analyzed platelet aggregation tests with different agonists (ADP, collagen, adrenalin) when normal platelets were exposed to serum with different concentrations of antiphospholipid antibodies. RESULTS: Results demonstrated a significant inhibition in adrenalin- and ADP-induced platelet aggregation curves (P < 0.05) in all antibody concentrations tested when compared to the control. The paradox between the prothrombotic state and the presence of autoantibodies that show anticoagulant activity in vitro was demonstrated in the literature, making it difficult to understand the pathophysiologic mechanism of the antiphospholipid syndrome. CONCLUSION: Results showed that anticardiolipin and anti-b2GPI antibodies-rich serum, both of which belonging to the IgG class, can interfere with platelet aggregation curves.

Urine storage under refrigeration preserves the sample in chemical, cellularity and bacteriuria analysis of ACS / Armazenamento da urina, sob refrigeração, preserva a amostra nas análises químicas, celularidade e bacteriuria no EAS

INTRODUCTION: The analysis of urine abnormal constituents and sediment (ACS) comprises tests of great diagnostic and prognostic value in clinical practice. When the analysis of ACS cannot be performed within two hours after collection, the sample must be preserved in order to avoid pre-analytical interferences. Refrigeration is the most applied technique due to its cost effectiveness. Moreover, it presents fewer inconveniences when compared to chemical preservation. However, changes in ACS may also occur in samples under refrigeration. OBJECTIVE: To analyze the influence of refrigeration at 2 to 8ºC on the storage of urine samples within 24 hours. MATERIAL AND METHOD: A total of 80 urine samples were selected from patients admitted at Universidade Federal de Juiz de Fora (UFJF) university hospital, which were tested for ACS at room temperature and stored under refrigeration for 6, 12 and 24 hours. RESULTS: The results showed that refrigeration proved to be effective when compared to samples kept at room temperature, inasmuch as the physical, chemical, microbial and cellularity features were preserved. Nevertheless, crystalluria was present after a 6- hour storage period. CONCLUSION: The tests revealed that cooling preserved cellularity and chemical characteristics of urine samples for up to 12 hours. Nonetheless, the precipitation of crystals was evident in this storage method. Thus, the possible consequences of storing urine samples for ACS test under these conditions should be included in the analysis report.

INTRODUÇÃO: A pesquisa de elementos anormais e sedimentoscopia na urina (EAS) compreende testes de grande valor diagnóstico e prognóstico na prática clínica. Quando a análise do EAS não puder ser realizada dentro de duas horas após a coleta da amostra, esta deve ser conservada para que interferências pré-analíticas sejam evitadas. A refrigeração é a técnica mais utilizada devido ao custo-benefício e por apresentar menos inconvenientes quando comparada com conservantes químicos. No entanto, alterações no EAS também podem ocorrer na amostra sob refrigeração. OBJETIVO: Analisar a influência da refrigeração entre 2 a 8ºC no armazenamento do EAS por um período de até 24 horas. MATERIAL E MÉTODO: Foram selecionadas 80 amostras de urina de pacientes internados no hospital da Universidade Federal de Juiz de Fora (UFJF) testadas para EAS, à temperatura ambiente, e armazenadas sob refrigeração em 6, 12 e 24 horas. RESULTADOS: Os resultados mostraram que a refrigeração foi eficaz quando comparada com amostras mantidas à temperatura ambiente, já que as características físicas, químicas, da celularidade e da microbiota da urina foram preservadas. No entanto, a cristalúria se fez presente desde as 6 horas de armazenamento. CONCLUSÃO: Os testes demonstraram que a refrigeração preservou as características químicas e a celularidade da urina por até 12 horas. No entanto, precipitações de cristais mostraram-se evidentes neste método de armazenamento. Dessa forma, a sugestão de se relatar no laudo as possíveis consequências dessa forma de armazenamento de urina para o EAS pode ser importante.

Avaliação da agregação plaquetária em presença de anticorpos antifosfolípides: anti-β2GP1 e anticardiolipina / Evaluation of platelet aggregation in the presence of antiphospholipid antibodies: anti-β2GP1 and anticardiolipin

INTRODUÇÃO: A síndrome antifosfolípide (SAF) é uma condição autoimune que apresenta fenômenos trombóticos arteriais e venosos de repetição além de complicações obstétricas. Sua patogênese está associada à presença de anticorpos antifosfolípides e/ou anti-β2 glicoproteína I (β2GPI) que aparentemente modificam o efeito anticoagulante da β2GPI. A dimerização da β2GPI induzida por anticorpos parece estar relacionada à indução da agregação plaquetária contribuindo para o estado trombofílico na SAF. OBJETIVOS: O presente trabalho objetiva demonstrar a influencia dos anticorpos antifosfolípides em testes de agregação plaquetária com diferentes agonistas (ADP, colágeno e adrenalina). MÉTODOS: Foram analisados testes de agregação de plaquetas normais com diferentes agonistas (ADP, colágeno, adrenalina) na presença de soro contendo anticorpos antifosfolípides em diferentes concentrações. RESULTADOS: As análises obtidas mostraram uma inibição significativa (P < 0,05) nas curvas de agregação plaquetária induzidas por ADP e adrenalina quando comparadas ao controle. O paradoxo entre o estado protrombótico e a presença de autoanticorpos que in vitro apresentam atividade anticoagulante foi demonstrado na literatura, dificultando o entendimento patofisiológico da síndrome antifosfolípide. CONCLUSÃO: Os resultados obtidos demonstraram que o soro rico em anticorpos anticardiolipina e anti-β2GPI, ambas da classe IgG, interferem em testes de curvas de agregação plaquetária.

INTRODUCTION: The antiphospholipid syndrome (APS) is an autoimmune condition characterized by recurrent arterial and venous thrombosis, besides obstetric complications. The pathogenesis is associated with the presence of antiphospholipid and/or anti-b2-glicoprotein I (anti-b2GPI) antibodies that appear to change the anticoagulant activity of b2GPI. Antibody-induced dimerization of b2GPI seems to be related to the induction of platelet aggregation, contributing to the development of thrombosis in APS. OBJECTIVES: The objective of the present study is to demonstrate the influence of antiphospholipid antibodies in platelet aggregation tests with different agonists (ADP, collagen, and adrenaline). METHODS: We analyzed platelet aggregation tests with different agonists (ADP, collagen, adrenalin) when normal platelets were exposed to serum with different concentrations of antiphospholipid antibodies. RESULTS: Results demonstrated a significant inhibition in adrenalin- and ADP-induced platelet aggregation curves (P < 0.05) in all antibody concentrations tested when compared to the control. The paradox between the prothrombotic state and the presence of autoantibodies that show anticoagulant activity in vitro was demonstrated in the literature, making it difficult to understand the pathophysiologic mechanism of the antiphospholipid syndrome. CONCLUSION: Results showed that anticardiolipin and anti-b2GPI antibodies-rich serum, both of which belonging to the IgG class, can interfere with platelet aggregation curves.

[Waist circumference measured before the 12th week of pregnancy: correlation with serum leptin levels]. / Circunferência abdominal avaliada antes da 12(a) semana de gestação: correlação com níveis séricos de leptina.

PURPOSE: To evaluate the correlation between maternal waist circumference measured before the 12th week of gestation and serum leptin levels during pregnancy, as well as to compare the leptin levels of women with and without abdominal obesity diagnosed in early pregnancy. METHODS: Prospective study including 40 pregnant women receiving low-risk prenatal care, older than 20 years, nonsmokers, with singleton pregnancies and without chronic disease. Waist circumference was measured before the 12th week and serum leptin levels were measured between the 9th and 12th, 25th and 28th and 34th and 37th weeks of gestation. According to waist circumference measurement, the cohort was divided into two groups: with and without abdominal obesity. The Mann-Whitney and χ(2) tests were used to assess the differences between groups. The Pearson correlation coeffient was used to assess the association between waist circumference and serum leptin levels during pregnancy. The level of significance was set at p<0.05. RESULTS: The mean weight and body mass index of patients with abdominal obesity (74.4±11.0 kg/28.9±4.1) was higher than that of patients without abdominal obesity (55.6±5.9 kg/21.1±2.4) (p=0.001). The mean leptin levels in pregnant patients with abdominal obesity (41.9±3.5 ng/mL) was higher than in patients without abdominal obesity (23.6±2.7 ng/mL) (p<0.0002). A positive correlation was obtained between the waist circumference measured during the same period and the mean serum leptin levels (r=0.7; p<0.0001). CONCLUSIONS: Waist circumference measured before the 12th week of pregnancy is a valid and simple method to predict the serum leptin levels throughout pregnancy. Pregnant women with abdominal obesity diagnosed before 12th week have higher mean serum leptin levels during pregnancy than those without abdominal obesity.

Circunferência abdominal avaliada antes da 12ª semana de gestação: correlação com níveis séricos de leptina / Waist circumference measured before the 12th week of pregnancy: correlation with serum leptin levels

OBJETIVOS: Avaliar a correlação entre a circunferência abdominal materna, medida antes da 12ª semana de gestação, e os níveis séricos de leptina durante a gravidez, bem como, comparar os níveis médios de leptina entre gestantes com e sem obesidade abdominal, diagnosticada no início da gestação. MÉTODOS: Estudo prospectivo incluindo 40 gestantes atendidas no pré-natal de baixo risco, superiores a 20 anos, não tabagistas, com gestação única, e sem doenças crônicas intercorrentes. A circunferência abdominal foi medida antes da 12ª semana, e os níveis séricos de leptina dosados entre a 9ª e a 12ª, a 25ª e a 28ª e entre a 34ª e a 37ª semanas de gestação. De acordo com a circunferência abdominal, a coorte foi dividida em dois grupos: com e sem obesidade abdominal. Os testes de Mann-Whitney e do χ² avaliaram as diferenças entre os grupos. A correlação de Pearson verificou a associação entre a circunferência abdominal e os níveis séricos de leptina durante a gestação. Considerou-se o valor de p<0,05. RESULTADOS: A média do peso e do índice de massa corpórea das pacientes com obesidade abdominal (74,4±11,0 kg/28,99±4,1) foi maior do que naquelas sem obesidade abdominal (55,6±5,9 kg/21,1±2,40) (p=0,001). A média dos níveis séricos de leptina no grupo das gestantes com obesidade abdominal (41,9±3,5 ng/mL) foi superior ao grupo das pacientes sem obesidade abdominal (23,6±2,7 ng/mL) (p<0,0002). Verificou-se, também, correlação entre a medida da circunferência abdominal e a média dos níveis séricos de leptina (r=0,7; p<0,0001). CONCLUSÕES: A circunferência abdominal medida antes da 12ª semana de gestação é um método válido e simples para se predizer os níveis séricos de leptina durante todo o período gestacional. Gestantes com obesidade abdominal diagnosticada antes da 12ª semana apresentam níveis médios de leptina sérica, durante a gravidez, superiores àquelas sem obesidade abdominal.

PURPOSE: To evaluate the correlation between maternal waist circumference measured before the 12th week of gestation and serum leptin levels during pregnancy, as well as to compare the leptin levels of women with and without abdominal obesity diagnosed in early pregnancy. METHODS: Prospective study including 40 pregnant women receiving low-risk prenatal care, older than 20 years, nonsmokers, with singleton pregnancies and without chronic disease. Waist circumference was measured before the 12th week and serum leptin levels were measured between the 9th and 12th, 25th and 28th and 34th and 37th weeks of gestation. According to waist circumference measurement, the cohort was divided into two groups: with and without abdominal obesity. The Mann-Whitney and χ² tests were used to assess the differences between groups. The Pearson correlation coeffient was used to assess the association between waist circumference and serum leptin levels during pregnancy. The level of significance was set at p<0.05. RESULTS: The mean weight and body mass index of patients with abdominal obesity (74.4±11.0 kg/28.9±4.1) was higher than that of patients without abdominal obesity (55.6±5.9 kg/21.1±2.4) (p=0.001). The mean leptin levels in pregnant patients with abdominal obesity (41.9±3.5 ng/mL) was higher than in patients without abdominal obesity (23.6±2.7 ng/mL) (p<0.0002). A positive correlation was obtained between the waist circumference measured during the same period and the mean serum leptin levels (r=0.7; p<0.0001). CONCLUSIONS: Waist circumference measured before the 12th week of pregnancy is a valid and simple method to predict the serum leptin levels throughout pregnancy. Pregnant women with abdominal obesity diagnosed before 12th week have higher mean serum leptin levels during pregnancy than those without abdominal obesity.