Involvement of GPR43 Receptor in Effect of Lacticaseibacillus rhamnosus on Murine Steroid Resistant Chronic Obstructive Pulmonary Disease: Relevance to Pro-Inflammatory Mediators and Oxidative Stress in Human Macrophages.

BACKGROUND: Cytokine storm and oxidative stress are present in chronic obstructive pulmonary disease (COPD). Individuals with COPD present high levels of NF-κB-associated cytokines and pro-oxidant agents as well as low levels of Nrf2-associated antioxidants. This condition creates a steroid-resistant inflammatory microenvironment. Lacticaseibacillus rhamnosus (Lr) is a known anti-cytokine in lung diseases; however, the effect of Lr on lung inflammation and oxidative stress in steroid-resistant COPD mice remains unknown. OBJECTIVE: Thus, we investigated the Lr effect on lung inflammation and oxidative stress in mice and macrophages exposed to cigarette smoke extract (CSE) and unresponsive to steroids. METHODS: Mice and macrophages received dexamethasone or GLPG-094 (a GPR43 inhibitor), and only the macrophages received butyrate (but), all treatments being given before CSE. Lung inflammation was evaluated from the leukocyte population, airway remodeling, cytokines, and NF-κB. Oxidative stress disturbance was measured from ROS, 8-isoprostane, NADPH oxidase, TBARS, SOD, catalase, HO-1, and Nrf2. RESULTS: Lr attenuated cellularity, mucus, collagen, cytokines, ROS, 8-isoprostane, NADPH oxidase, and TBARS. Otherwise, SOD, catalase, HO-1, and Nrf2 were upregulated in Lr-treated COPD mice. Anti-cytokine and antioxidant effects of butyrate also occurred in CSE-exposed macrophages. GLPG-094 rendered Lr and butyrate less effective. CONCLUSIONS: Lr attenuates lung inflammation and oxidative stress in COPD mice, suggesting the presence of a GPR43 receptor-dependent mechanism also found in macrophages.

Lactobacillus rhamnosus Modulates Lung Inflammation and Mitigates Gut Dysbiosis in a Murine Model of Asthma-COPD Overlap Syndrome.

The asthma-COPD overlap syndrome (ACOS) presents lung inflammation similar to both asthma and chronic obstructive pulmonary disease (COPD). Due to the immune response between the lung and gut, it is possible that ACOS individuals present gut dysbiosis. Due to therapeutic limitations in ACOS, Lactobacillus rhamnosus (Lr) have received attention once Lr has been effective in asthma and COPD. However, there is no data about the Lr effect on both lung inflammation and gut dysbiosis in ACOS. Thus, our study investigated the Lr effect on lung inflammation, bronchoconstriction, airway remodeling, and gut dysbiosis in the murine ACOS model. Treated mice with Lr were exposed to HDM and cigarette smoke to induce ACOS. Sixty days after ACOS induction, mice were euthanized. Lung inflammation was evaluated in leukocytes in bronchoalveolar lavage fluid (BALF), airway remodeling, cytokine secretion, and transcription factor expression in the lung. The gut microbiota was assayed by 16S mRNA sequencing from a fecal sample. Leukocyte population, bronchial hyperreactivity, pro-inflammatory cytokines, and airway remodeling were attenuated in Lr-treated ACOS mice. Likewise, IL-4, IL-5, and IL-13, STAT6 and GATA3, as well as IL-17, IL-21, IL-22, STAT3, and RORÉ£t were reduced after Lr. In addition, IL-2, IL-12, IFN-γ, STAT1, and T-bet as well as IL-10, TGF-ß, STAT5, and Foxp3 were restored after the Lr. Firmicutes was reduced, while Deferribacteres was increased after Lr. Likewise, Lr decreased Staphylococcus and increased Mucispirillum in ACOS mice. Lr improves fecal bacterial ß-diversity. Our findings show for the first time the Lr effect on lung inflammation and gut dysbiosis in murine ACOS.

Higher systemic inflammatory status and cardiovascular risk associated with Charcot arthropathy unrelated to infection or extremity amputation.

Purpose: People with diabetes and Charcot arthropathy have higher mortality than people with diabetes without this complication. Are the causes of this higher mortality exclusively infectious or of a cardiovascular origin? We aimed to study aspects related to cardiovascular risk and inflammation in a population of people with type 2 diabetes with and without Charcot arthropathy. Methods: A cross-sectional study was performed in people with diabetes and Charcot Eickenholtz III arthropathy, matched for sex and age, with two groups of people with diabetes without Charcot arthropathy with and without peripheral sensory-motor neuropathy, in the absence of active infection. All participants underwent clinical and laboratory evaluation at the time of the interview, and their cardiovascular risk was calculated according to the United Kingdom Prospective Diabetes Study (UKPDS) calculator. Results: We evaluated 69 people with type 2 diabetes (21 with Charcot arthropathy, 24 with diabetic peripheral neuropathy and 24 without this neuropathy), with a mean age between 57 and 61 years and with a diabetes duration of more than 10 years. Patients with Charcot arthropathy had a longer duration of diabetes; more frequency of dyslipidemia; and higher levels of uric acid, microalbuminuria and C-reactive protein than the other two groups. A total of 73.9% of the people evaluated had high or very high cardiovascular risk according to the UKPDS score. Conclusion: The people with type 2 diabetes presenting Charcot arthropathy had greater values of systemic inflammatory parameters, even in the chronic phase of the complication, with a higher risk of mortality unrelated to infections.

Influence of thyroid hormone in the expression of the marker pro-apoptosis BID, in spite of the predominance of anti-apoptosis activation in intratiroidal lymphocytic infiltration in Hashimoto's thyroiditis.

Cell destruction in Hashimoto's thyroiditis (HT) involves autoantibodies and cytotoxic T lymphocytes. Thyrocytes maintenance occurs by pro-apoptotic, anti-apoptotic and cell proliferation balance. OBJECTIVES: To characterize factors related to the mechanisms of apoptosis and cell proliferation in thyroid cells and intrathyroid lymphocytic infiltrate in HT. METHODS: We assessed lymphocytic infiltrate and thyroid cells from HT and normal thyroid by immunohistochemical analysis of cell proliferation (Ki-67), antiproliferation (p27Kip1), pro-apoptosis (Fas, Fas-ligand, BID) and anti-apoptosis (MCL-1, BCL2) markers. RESULTS: Lymphocytic infiltrate presented BCL2 and MCL-1 higher expression, Ki-67 and p27kip1 balance. Thyrocytes exhibited Fas and FasL balance, higher BID expression; MCL-1, BCL-2, Ki-67 similar to the normal thyroid. T4 and higher lymphocytes BID expression were associated. CONCLUSIONS: In lymphocytic infiltrate predominated anti-apoptosis in relation to pro-apoptosis except for BID. Thyrocytes presented pro-apoptosis and anti-apoptosis balance and cell proliferation similar to normal thyroid. T4-associated BID expression in HT lymphocytes suggests the influence of thyroid hormone as a signal to up-regulate the BID pro-apoptotic protein and thus increase lymphocytic apoptosis rates.

Ocular Manifestations and Neuropathy in Type 2 Diabetes Patients With Charcot Arthropathy.

Objective: Diabetes can affect the eye in many ways beyond retinopathy. This study sought to evaluate ocular disease and determine any associations with peripheral neuropathy (PN) or cardiac autonomic neuropathy (CAN) in type 2 diabetes (T2D) and Charcot arthropathy (CA) patients. Design: A total of 60 participants were included, 16 of whom were individuals with T2D/CA, 21 of whom were individuals with T2D who did not have CA, and 23 of whom were healthy controls. Ocular surface evaluations were performed, and cases of dry eye disease (DED) were determined using the Ocular Surface Disease Index (OSDI) questionnaire, ocular surface staining, Schirmer test, and Oculus Keratograph 5M exams. All variables were used to classify DED and ocular surface disorders such as aqueous deficiency, lipid deficiency, inflammation, and ocular surface damage. Pupillary and retinal nerve fiber measurements were added to the protocol in order to broaden the scope of the neurosensory ocular evaluation. PN and CAN were ascertained by clinical examinations involving the Neuropathy Disability Score (for PN) and Ewing's battery (for CAN). Results: Most ocular variables evaluated herein differed significantly between T2D patients and controls. When the controls were respectively compared to patients with T2D and to patients with both T2D and CA, they differed substantially in terms of visual acuity (0.92 ± 0.11, 0.73 ± 0.27, and 0.47 ± 0.26, p=0.001), retinal nerve fiber layer thickness (96.83 ± 6.91, 89.25 ± 10.44, and 80.37 ± 11.67 µm, p=0.03), pupillometry results (4.10 ± 0.61, 3.48 ± 0.88, and 2.75 ± 0.81 mm, p=0.0001), and dry eye symptoms (9.19 ± 11.71, 19.83 ± 19.08, and 24.82 ± 24.40, p=0.03). DED and ocular surface damage also differed between individuals with and without CA, and were associated with PN and CAN. Conclusion: CA was found to be significantly associated with the severity of ocular findings. DED in cases of CA was also associated with PN and CAN. These findings suggest that intrinsic and complex neurosensory impairment in the eyes, peripheral sensory nerves, and the autonomic nervous system are somehow connected. Thus, a thorough ocular evaluation may be useful to highlight neurological complications and the impact of diabetes on ocular and systemic functions and structures.

Induction, biochemical trait and phytochemical screening of calluses of Amburana cearensis (Allemão) A. C. Smith

Amburana cearensisis an arboreal legume of the Fabaceaefamily,with high phytotherapic and medicinal potential due the presence of secondary metabolites. The objective of this study was to evaluate the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-2,5,6-trichloro-2-pyridinecarboxylic acid (picloram) on the in vitroinduction of callogenesis of A. cearensisand analyze the biochemical and phytochemical potential of these calluses. For callus induction, leaf and cotyledon segments were used as explants, which were inoculated in woody plant medium (WPM) supplemented with different concentrations of 2,4-D (0, 5, 10, 20, 40 μM) or picloram (0, 5, 10, 20, 40, 80 μM). The callus growth curve was estimated based on fresh weight, measured at 7-day intervals until 28 days after inoculation. The calluses were analyzed by biochemical tests to quantify the reducing sugars and total proteins. Phytochemical screening and high-performance liquid chromatography were performed to establish the phytochemical profile of extracts from calluses. The concentrations of 21.94 μMand 26.46 μMof 2,4-Dinduced the greatest formation of compact and friable calluses from the leaf and cotyledon segments, respectively. The growth curve had two distinct phases(lag and exponential) for both types of calluses evaluated. The maximum levels of reducing sugars and total proteins in the calluses from leaf and cotyledon segments were obtained on the day of inoculation and after 28 days of cultivation, respectively. The results of the phytochemical analysis identified the presence of coumarin in all the extracts evaluated, this secondary metabolite has high pharmacological potential.

Value of Apoptotic, Antiapoptotic, and Cell Proliferation Markers in the Treatment of Graves' Disease.

To better understand the genesis of autoimmunity in Graves' disease (GD), it is essential to study the mechanism of apoptosis and cell proliferation in thyroid cells and intrathyroidal lymphocytic infiltrate of GD patients. Methods. A cross sectional, observational study performed by evaluating histopathological samples of thyroidectomy products from GD patients using immunohistochemistry. New histological sections were prepared for immunohistochemical analysis with markers of cell proliferation, antiproliferation, apoptosis, and antiapoptosis. Results. Patients with GD who underwent radioiodine therapy (RIT) had a lower lymphocytic expression level of p27Kip1, and those who took beta-blockers had higher expression levels of BID (BH3-interacting domain) and a lower Ki-67 expression level in thyrocytes than those who did not. The association of a shorter diagnostic time with a lower expression level of MCL-1 in thyroid cells suggests that the hyperthyroid state was related to a lower antiapoptotic effect on thyrocytes. In comparison to patients with GD not using antithyroid drugs (ATD), we found a lower expression level of BID in lymphocytes for those who used ATD. Conclusion. In GD, the hyperthyroid state was associated with a lower antiapoptotic effect on thyroid cells. RIT, beta-blockers, and thionamide act by stimulating apoptosis of thyrocytes by intrathyroidal lymphocytes.

Mulheres com síndromes hipertensivas / Women with hypertensive syndromes

Objetivo: descrever o perfil obstétrico de mulheres com síndrome hipertensiva. Método: estudo quantitativo, exploratório, descritivo, retrospectivo e de delineamento documental, com 196 prontuários de pacientes com síndromes hipertensivas, no período de novembro de 2016 a maio de 2017. Para a coleta dos dados, foi utilizado um formulário. Em seguida, eles foram agrupados e organizados em uma planilha Excel e, posteriormente, convertidos em tabela. Resultados: a faixa etária predominante das pacientes estava entre os 16 aos 30 anos, com 68,88%; em relação ao número de partos, 55,61% eram multíparas; sobre o acompanhamento do pré-natal, 87,76% das gestantes tiveram acima de seis consultas; 89,80% das gestações eram únicas e 10,20%, gemelares; 57,27% dos recém-nascidos eram do sexo masculino e 42,73% eram do sexo feminino, apresentando 0,45% dos recém-nascidos com APGAR menor que sete no quinto minuto de vida e 99,55% com APGAR maior que sete. Conclusão: o estudo mostrou-se necessário para identificar fatores de risco que auxiliam na qualificação da assistência à mulher.(AU)

Muscle activation during resistance training with no external load - effects of training status, movement velocity, dominance, and visual feedback.

OBJECTIVES: To explore the acute effects of training status, movement velocity, dominance, and visual feedback on muscle activation and rating of perceived exertion (RPE) during resistance training with no external load (no-load resistance training; NLRT). METHODS: Thirty-three men (17 untrained and 16 trained), performed elbow flexions in four NLRT sessions: 1) slow velocity with EMG visual feedback, 2) slow velocity without EMG visual feedback, 3) fast velocity with EMG feedback, and 4) fast velocity without EMG feedback. RPE was measured using the Borg Discomfort scale. EMG for the biceps and triceps were recorded for both arms. RESULTS: EMG feedback had no influence on RPE. The peak and mean EMG values were not different for the biceps (93.8±11.5% and 50±13.1%) and triceps (93.7±23.9% and 49.6±16.2%). The results revealed a difference in the training status, with higher peak EMG for untrained than for trained participants (96.9±20% vs. 90.2±15.6%). However the values for mean EMG were not different between the untrained and trained (50.3±15.7% vs. 49.2±13.7%) participants. There was no difference in the peak (92.8±19% vs. 94.7±20.4%) and mean (49.8±15.0% vs. 49.7±14.5%) EMG values for the dominant and non-dominant sides. Peak EMG values were not different between faster and slower velocities (93.6±19.6% and 93.9±17.8%). However, mean EMG was higher for slower (50.5±14.4%) than for faster (48.5±15.4%) velocities. The peak and mean EMG during contractions with (93.3±17.5% and 49.5±14.1%) and without visual feedback (94.2±19.9% and 50±15.4%) were not significantly different. CONCLUSION: NLRT produces high levels of muscle activation independent of training, status, dominance, movement velocity, and visual feedback.

Omega-3 Fatty Acids: Possible Neuroprotective Mechanisms in the Model of Global Ischemia in Rats.

Background. Omega-3 (ω3) administration was shown to protect against hypoxic-ischemic injury. The objectives were to study the neuroprotective effects of ω3, in a model of global ischemia. Methods. Male Wistar rats were subjected to carotid occlusion (30 min), followed by reperfusion. The groups were SO, untreated ischemic and ischemic treated rats with ω3 (5 and 10 mg/kg, 7 days). The SO and untreated ischemic animals were orally treated with 1% cremophor and, 1 h after the last administration, they were behaviorally tested and euthanized for neurochemical (DA, DOPAC, and NE determinations), histological (Fluoro jade staining), and immunohistochemical (TNF-alpha, COX-2 and iNOS) evaluations. The data were analyzed by ANOVA and Newman-Keuls as the post hoc test. Results. Ischemia increased the locomotor activity and rearing behavior that were partly reversed by ω3. Ischemia decreased striatal DA and DOPAC contents and increased NE contents, effects reversed by ω3. This drug protected hippocampal neuron degeneration, as observed by Fluoro-Jade staining, and the increased immunostainings for TNF-alpha, COX-2, and iNOS were partly or totally blocked by ω3. Conclusion. This study showed a neuroprotective effect of ω3, in great part due to its anti-inflammatory properties, stimulating translational studies focusing on its use in clinic for stroke managing.

Citrus rootstocks influence the population densities of pest mites / Porta-enxertos cítricos influenciam a densidade populacional de ácaros fitófagos

Citrus are attacked by pest mites such as the citrus rust mite Phyllocoptruta oleivora (Ashmead) (Acari: Eriophyidae) and the spider mite Tetranychus mexicanus (McGregor) (Acari: Tetranychidae). However, little is known on citrus rootstocks influencing pest mites. We aimed to evaluate the influence of rootstocks on population densities of pest mites on the sweet oranges 'Pera CNPMF D-6' and 'Valencia Tuxpan' throughout time. Adults of both mite species were monthly counted during 19 months from June 2011 to February 2013. Rootstocks influenced the populations of pest mites, since lower densities of P. oleivora were found on 'Pera CNPMF D-6' sweet orange grafted on the hybrid TSKC × CTTR - 002 and on 'Swingle' citrumelo in comparison with the hybrid LVK × LCR - 010, 'Red' rough lime and 'Santa Cruz' rangpur lime as rootstocks. Similarly, lower densities of T. mexicanus were found on 'Valencia Tuxpan' sweet orange grafted on the hybrid HTR-051 in comparison to 'Indio' citrandarin, 'Sunki Tropical' mandarin and LVK × LCR - 010 as rootstocks. We concluded that densities of the mites P. oleivora and T. mexicanus on the sweet oranges 'Pera CNPMF D-6' and 'Valencia Tuxpan' were affected in some periods of the year by some rootstocks, suggesting influence of some genotypes on these pests.

Os citros são atacados por ácaros-praga como o ácaro-da-falsa-ferrugem Phyllocoptruta oleivora (Ashmead) (Acari: Eriophyidae) e Tetranychus mexicanus (McGregor) (Acari: Tetranychidae). No entanto, pouco é conhecido sobre o efeito de porta-enxertos sobre populações de pragas. O objetivo deste trabalho foi avaliar a influência de porta-enxertos nas densidades populacionais de ácaros-praga nas laranjeiras 'Pera CNPMF D-6' e 'Valência Tuxpan' ao longo do tempo. Adultos das espécies de ácaro citadas foram quantificados mensalmente, durante 19 meses, de junho de 2011 a fevereiro de 2013. Os porta-enxertos influenciaram as populações dos ácaros-praga, uma vez que menores densidades de P. oleivora foram encontradas em laranjeira 'Pera CNPMF D-6', enxertada no híbrido TSKC × CTTR - 002 e no citrumelo 'Swingle', em comparação com o que se observou em relação aos porta-enxertos LVK × LCR - 010, limoeiro 'Rugoso vermelho' e limoeiro 'Cravo Santa Cruz'. Similarmente, menores densidades de T. mexicanus ocorreram em laranjeira 'Valência Tuxpan' enxertada no híbrido HTR-051, em comparação com o que se deu em relação aos porta-enxertos citrandarin 'Indio', tangerineira 'Sunki Tropical' e LVK × LCR - 010. Concluiu-se que as densidades dos ácaros P. oleivora e T. mexicanus nas laranjeiras 'Pera CNPMF D-6' e 'Valencia Tuxpan' foram afetadas pelo porta-enxerto em determinados períodos do ano, sugerindo influência de alguns genótipos sobre essas pragas.

Studies on intracellular delivery of carboxyl-coated CdTe quantum dots mediated by fusogenic liposomes.

The use of Quantum Dots (QDs) as fluorescent probes for understanding biological functions has emerged as an advantageous alternative over application of conventional fluorescent dyes. Intracellular delivery of QDs is currently a specific field of research. When QDs are tracking a specific target in live cells, they are mostly applied for extracellular membrane labeling. In order to study intracellular molecules and structures it is necessary to deliver free QDs into the cell cytosol. In this work, we adapted the freeze and thaw method to encapsulate water dispersed carboxyl-coated CdTe QDs into liposomes of different compositions, including cationic liposomes with fusogenic properties. We showed that labeled liposomes were able to fuse with live human stem cells and red blood cells in an endocytic-independent way. We followed the interactions of liposomes containing QDs with the cells. The results were minutely discussed and showed that QDs were delivered, but they were not freely diffused in the cytosol of those cells. We believe that this approach has the potential to be applied as a general route for encapsulation and delivery of any membrane-impermeant material into living cells.