Gram-negative bacteria associated with a dominant arboreal ant species outcompete phyllosphere-associated bacteria species in a tropical canopy.

Ants have efficient and well-studied social immunity mechanisms, which prevent the colony contamination. Little is known about how workers keep their outside territory clear of diseases. We investigated the interactions between Azteca chartifex ants, their associated bacteria and bacteria on the phyllosphere of Byrsonima sericea trees, comparing plants patrolled and not by the ants. The hypothesis is that bacteria associated with the worker's exoskeleton may outcompete the leaf bacteria. Culturable bacteria were isolated from ants, from the main and satellite nests, and from phyllosphere of B. sericea taken from trees that had A. chartifex nests and from trees without nests. The isolates were grouped by Gram guilds and identified at the genus level. There was a higher percentage of Gram-negative isolates in the ants and on the leaves patrolled by them. There was a higher growth rate of ant bacteria from the main nest compared to those found in ants from the satellite nests. The most representative genus among ant isolates was Enterobacter, also found on leaves patrolled by ants. Under favourable in vitro conditions, A. chartifex Gram-negative bacteria outcompete leaf bacteria by overgrowth, showing a greater competition capacity over the Gram-positive bacteria from leaves with no previous interaction with ants in the field. It was demonstrated that ants carry bacteria capable of outcompeting exogenous bacteria associated with their outside territory. The leaf microbiota of a patrolled tree could be shaped by the ant microbiota, suggesting that large ant colonies may have a key role in structuring canopy plant-microbe interactions.

The Importance of Forest Simplification and Litter Disturbance in Defining the Assembly of Ground-Foraging Ants.

Currently, we are facing many ecosystem changes derived from years of anthropogenic disturbances. Habitat simplification stands out among human-derived impacts, due to its detrimental effects on vegetation structure and associated biota. Here, we assessed the effects of litter disturbance and forest simplification on a tropical ground-foraging ant community. To do that, we tested whether ant richness will be negatively affected by litter disturbance and habitat simplification. Additionally, we tested whether litter disturbance affects the time of resource discovery and dominance, and if so, whether its effects are intensified by forest simplification. This study occurred at Rio Doce State Park, a preserved area of Atlantic Forest in Southeastern Brazil. We experimentally simulated litter disturbance by removing the leaf litter and superficial soil layer in a mahogany monoculture forest and preserved Atlantic Forest. We sampled ants using paired-mixed baits of protein and carbohydrate in 12 points, half of them in each forest type. As expected, we found higher richness in the preserved and non-disturbed forest. Moreover, resource discovery was faster in disturbed monoculture, but bait dominance was higher in the undisturbed preserved forest. Litter heterogeneity seems to play an important role in determining ant dispersion and intra-specific communication, as we observed that litter disturbance impacts were strengthened by forest simplification. Our results highlight the efficiency of ground-foraging ants as bioindicators of disturbance and habitat quality. Moreover, our study indicates how distinct types of disturbances can act synergistically, changing the assembly of associated biota.

Estudo químico e da atividade biológica cardiovascular do óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L.Burtt & R.M.Sm. em ratos / Phytochemistry and cardiovascular biological activity of the essential oil from leaves of Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. in rats

A espécie vegetal Alpinia zerumbet (Pers.) B.L.Burtt & R.M. Sm. é popularmente empregada para o tratamento de diversas enfermidades, entre elas a hipertensão. Avaliar a composição química, a atividade antihipertensiva e ação na hipertrofia cardíaca do óleo essencial das folhas de Alpinia zerumbet (OEAZ) em ratos foram os objetivos deste estudo. O OEAZ, obtido por hidrodestilação em aparelho Clevenger, teve sua composição química analisada em cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Foram identificados 14 constituintes, sendo terpinen-4-ol (37,45 por cento) o majoritário, seguido pelos óxido de cariofileno (7,56 por cento), trans-hidrato de sabineno (6,61 por cento) e 1,8-cineol (4,02 por cento). A avaliação cardiovascular foi feita após o tratamento crônico de ratos espontaneamente hipertensos (SHR) e seus respectivos controles, ratos Wistar-Kyoto (WKY). Os dados hemodinâmicos revelaram redução da pressão arterial média (PAM) no grupo tratado (SHRP: 160 ± 7 mm Hg; p<0,01) em relação ao não tratado (SHR: 180 ± 5 mm Hg). A relação entre peso do ventrículo esquerdo e peso corporal (VE/PC) do SHRP (2,50 ± 0,03 mg g-1; p<0,01) mostrou-se inferior ao SHR (2,61 ± 0,01 mg g-1), confirmando a redução da hipertrofia cardíaca (HC). Os dados de PAM e VE/PC dos animais SHRP foram estatisticamente diferentes quando comparados com os ratos controle (WKY: 116 ± 2 mm Hg e WKYP: 119 ± 4 mm Hg; p<0,05; WKY: 2,15 ± 0,04 mg g-1 e WKYP: 2,17 ± 0,04 mg g-1 ; p<0,01), indicando não ter havido normalização dos mesmos. Conclui-se que o tratamento crônico com OEAZ foi capaz de determinar redução, mas não a normalização, da PAM e da HC de ratos SHR, provavelmente pela presença dos componentes terpinen-4-ol e 1,8-cineol. Estudos com doses maiores ou período de tratamento superior são necessários para avaliar a possibilidade de o OEAZ normalizar os parâmetros analisados (PAM e HC).

Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. is traditionally employed to treat several diseases such as hypertension. The aim of this study was to evaluate the chemical composition, the anti-hypertensive activity and the capacity to reduce cardiac hypertrophy of the essential oil of A. zerumbet leaves (EOAZ) in rats. EOAZ was obtained through hydrodistillation in Clevenger apparatus and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Several constituents (14) were identified, terpen-4-ol (37.45 percent) being the major component, followed by caryophyllene oxide (7.56 percent), trans-sabinene hydrate (6.61 percent) and 1,8-cineol (4.02 percent). The cardiovascular effect was investigated after chronic treatment with spontaneously hypertensive rats (SHR) and their respective controls, Wistar-Kyoto rats (WKY). The treated group showed a lower mean arterial pressure (MAP) (SHRP: 160 ± 7 mm Hg; p<0.01) than the untreated group (SHR: 180 ± 5 mm Hg). The ratio of left ventricle-to-body weight (LV/BW) for SHRP was lower (2.504 ± 0.03 mg g-1; p<0.01) than that for SHR (2.162 ± 0.01 mg g-1), confirming the cardiac hypertrophy (CH) reduction. There were significant differences in MAP and CH between SHRP animals and control rats (WKY: 116 ± 2 mm Hg and WKYP: 119 ± 4 mm Hg; p<0.05. WKY: 2.152 ± 0.04 mg g-1 and WKYP: 2.168 ± 0.04 mg g-1; p<0.01), indicating that these values were not normalized. Those data showed that the chronic treatment with EOAZ reduces MAP and CH in SHR probably due to the presence of the compounds terpinen-4-ol and 1,8-cineol. Studies with higher doses or longer treatment periods are necessary to evaluate whether EOAZ can reduce the analyzed parameters (MAP and CH) to normal values.

Efeito da administração do Allium sativum sobre as alterações cardiovasculares de ratos Wistar com infarto do miocárdio. / Effects of consumption of Allium sativum on the cardiovascular abnormalities observed in Wistar rats following cardiac infarction.

O alho (Allium sativum) apresenta várias ações benéficas ao sistema circulatório, tais como diminuição dos níveis de colesterol total, LDL-colesterol e da pressão arterial, além de efeito antioxidante. O objetivo deste estudo foi avaliar o efeito da administração do Allium sativum sobre as alterações da hemodinâmica cardiovascular e estruturais macroscópicas do coração de animais com infarto induzido experimentalmente. Ratos Wistar foram tratados, previamente e após indução do infarto, com homogeneizado de alho na dose de 125mg/Kg/dia durante 21 dias, por via oral (uma semana antes e duas depois do procedimento de infarto). Os grupos controle passaram por cirurgia fictícia (SHAM). Os animais foram divididos em grupos controles e infartados com (SHAMT, INFT; respectivamente) ou sem (SHAM, INF; respectivamente) tratamento com alho. Houve redução da hipertrofia do ventrículo direito (INF=0,75±0,05 vs. INFT=0,61±0,03 mg/Kg; p<0,01), da área de infarto (INF=29,7±4,8% vs. INFT=20,4±1,4%; p<0,05) e regularização dos níveis de pressão arterial sistólica (PAS; INF=100±8 vs. INFT=127±7 mmHg; p<0,05) e média (PAM; INF=94±4 vs. INFT=110±6 mmHg; p<0,01) dos animais INFT comparados com os INF. Houve um menor número de animais mortos após o procedimento de infarto no grupo INFT em relação ao grupo INF (20%, n=2; 45,5%, n=5; respectivamente). Esses achados indicam que o alho tem um importante papel na prevenção e no controle de alterações cardiovasculares, uma vez que houve redução do número de mortes pós-infarto e melhor perfil cardiovascular dos animais INFT.

Garlic (Allium sativum) has several beneficial effects on the cardiovascular system, such as reductions of the levels of total cholesterol, LDL-cholesterol and blood pressure, besides acting as an antioxidant. The aim of this study was to assess the effects of administering Allium sativum on changes in the cardiovascular hemodynamics and macroscopic structure that occur in the hearts of animals with experimentally induced cardiac infarction. Male Wistar rats were treated with homogenized garlic at a dose of 125mg/kg b.w./day for 21 days, given orally for one week before and two weeks after the procedure to induce myocardial infarction. The control group was subjected to a fictitious surgery (SHAM). The animals were divided into control and infarcted groups, treated (SHAMT, INFT) or untreated (SHAM, INF) with garlic. There were reductions in right ventricular hypertrophy (INF=0.75±0.05 vs. INFT=0.61±0.03 mg.kg-1; p<0.01) and infarcted area (INF=29.7±4.8 % vs. INFT=20.4±1.4 %; p<0.05) and regularization of the levels of systolic (SAP; INF=100±8 vs. INFT=127±7 mm Hg; p<0.05) and mean arterial pressure (MAP; INF=94±4 vs. INFT=110±6 mm Hg; p<0.01) in the INFT animals, compared to the INF group. Fewer animals died after the cardiac infarction procedure in the group INFT than in INF (20%, n=2; 45.5%, n=5; respectively). These findings suggest that garlic can have an important role in the prevention and control of cardiovascular abnormalities, since there was a reduction in the number of post-infarction deaths and an improvement of the cardiovascular profile in the INFT animals.

Effect of enalapril treatment on the sensitivity of cardiopulmonary reflexes in rats with myocardial infarction.

1. The aim of the present study was to evaluate the effect of treatment with enalapril on the sensitivity of cardiopulmonary reflexes 30 days after myocardial infarction in Wistar rats. 2. Animals were divided into four groups: (i) sham operated, receiving vehicle (SHAM); (ii) infarcted, receiving vehicle (0.9% NaCl; INF); (iii) sham operated, receiving enalapril (SHAME); and (iv) infarcted, receiving enalapril (INFE). 3. Enalapril was administered at a dose of 10 mg/kg per day. Serotonin (4-32 microg/kg, i.v.) was administered in order to activate the Bezold-Jarisch reflex, which was estimated as the percentage of reduction in heart rate. 4. The volume-sensitive cardiopulmonary reflex was induced by saline overload and evaluated as the percentage increase in sodium and volume renal excretion. At the end of the experiments, rats were killed and hearts excised to estimate the size of the infarction. The weight of the kidneys, lungs, liver and cardiac chambers as ratios of bodyweight was used to estimate the extent of hypertrophy. 5. The results showed an impairment in the sensitivity of the cardiopulmonary reflexes in the INF group compared with the SHAM and SHAME groups. We observed right ventricle and pulmonary hypertrophy, a reduction in mean and systolic arterial pressure and an increase in heart rate in INF animals. In the INFE group, nearly all the parameters were normal compared with the INF group, except for systolic arterial pressure, which was only partially improved. 6. The main finding of the present study was that treatment with enalapril normalized the sensitivity of the cardiopulmonary reflexes, which could be due, in part, to the reduction of cardiac hypertrophy. The present study provides information about the beneficial effects of the angiotensin-converting enzyme inhibitors by normalizing the cardiopulmonary reflexes involved with the regulation of volume and sodium, as well as control of arterial pressure and heart rate in infarcted animals.

Enhanced beta-adrenergic response in rat papillary muscle by inhibition of inducible nitric oxide synthase after myocardial infarction.

AIM: Myocardial infarction (MI) induces a progressive ventricular remodelling leading to a contractility depression. During the acute phase of MI inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production increases in the heart. The aim of this study was to investigate the role of iNOS in the left ventricular contractility at 3 days after MI. METHODS: Wistar rats were divided into: sham operated (SHAM, n = 23), infarction (INF, n = 18); sham operated plus the iNOS inhibitor, S-methylisothiourea (SMT) 5 mg kg(-1) day(-1), i.p. treatment (SHAM-SMT, n = 26) and infarction plus SMT (INF-SMT, n = 22). Concentration-response curves for isoprenaline, Ca(2+) and frequency-force curve were studied in isolated papillary muscle from left ventricle. RESULTS: After 3 days infarct area was similar between groups. SMT treatment reduced the time to peak tension during frequency-force curve in the infarct group (SHAM = 63 +/- 3; SHAM-SMT = 71 +/- 3; INF = 90 +/- 4; INF-SMT = 79 +/- 4 ms, P < 0.05) and increased the maximal response to isoprenaline (SHAM = 0.93 +/- 0.11; SHAM-SMT = 1.13 +/- 0.1; INF =0.84 +/- 0.16; INF-SMT = 1.49 +/- 0.15 g mm(-2), P < 0.05). The response to Ca(2+) was equally reduced in the INF and INF-SMT groups. SMT treatment did not change the reduced post-rest potentiation performed by INF group, but attenuated the plasma nitrite and nitrate (NOx) levels in the INF group without any haemodynamic effect. CONCLUSION: These finding suggest that at 3 days after MI the iNOS modulates the isolated papillary muscle response to isoprenaline and its inhibition improves the beta-adrenergic inotropic responses.

Effects of mercury on the contractile activity of the right ventricular myocardium.

The increase in right ventricular systolic pressure observed in vivo after the administration of mercury opposes to the idea that the metal depresses the cardiac pump performance. We then investigated the effects of HgCl(2) (0.1 to 2.5 microM) on the contractile activity of the right ventricular myocardium, measuring isometric and tetanic contractions of right ventricular isolated strips, right ventricular isovolumic systolic and diastolic pressures, and the coronary perfusion pressure (0.03 to 3 microM) in constant-flow Langendorff-perfused rat hearts. The results presented here suggest that the acute effects of mercury on the right ventricular myocardium are distinct. When isolated strips of right ventricular wall are used, the contractile depression produced by mercury is manifested. However, when mercury is administered to isolated perfused hearts or in vivo this depressant effect is not revealed. The possible reasons for this behavior are the increased coronary perfusion pressure, which promotes a positive inotropic effect, manifested during the infusion of increasing concentrations of mercury, or the putative stretch of the ventricular fibers, which might cause the increment of diastolic pressure. An interesting finding is that the mechanical activity of the preparations, in which mercury is administered via coronary circulation, is not depressed and, even more, it can increase systolic pressure. However, the nature of this protective effect of coronary circulation cannot be explained by the results presented here.

Effects of small doses of ouabain on the arterial blood pressure of anesthetized hypertensive and normotensive rats

Ouabain increases vascular resistance and may induce hypertension by inhibiting the Na+ pump. The effects of 0.18 and 18 æg/kg, and 1.8 mg/kg ouabain pretreatment on the phenylephrine (PHE; 0.1, 0.25 and 0.5 æg, in bolus)-evoked pressor responses were investigated using anesthetized normotensive (control and uninephrectomized) and hypertensive (1K1C and DOCA-salt treated) rats. Treatment with 18 æg/kg ouabain increased systolic and diastolic blood pressure in all groups studied. However, the magnitude of this increase was larger for the hypertensive 1K1C and DOCA-salt rats than for normotensive animals, while the pressor effect of 0.18 æg/kg ouabain was greater only in DOCA-salt rats. A very large dose (1.8 mg/kg) produced toxic effects on the normotensive control but not on uninephrectomized or 1K1C rats. Rat tail vascular beds were perfused to analyze the effects of 10 nM ouabain on the pressor response to PHE. In all animals, 10 nM ouabain increased the PHE pressor response, but this increase was larger in hypertensive DOCA-salt rats than in normotensive and 1K1C rats. Results suggested that a) increases in diastolic blood pressure induced by 18 æg/kg ouabain were larger in hypertensive than normotensive rats; b) in DOCA-salt rats, smaller ouabain doses had a stronger effect than in other groups; c) hypertensive and uninephrectomized rats were less sensitive to toxic doses of ouabain, and d) after treatment with 10 nM ouabain isolated tail vascular beds from DOCA-salt rats were more sensitive to the pressor effect of PHE than those from normotensive and 1K1C hypertensive rats. These data suggest that very small doses of ouabain, which might produce nanomolar plasma concentrations, enhance pressor reactivity in DOCA-salt hypertensive rats, supporting the idea that endogenous ouabain may contribute to the increase and maintenance of vascular tone in hypertension

Effects of small doses of ouabain on the arterial blood pressure of anesthetized hypertensive and normotensive rats.

Ouabain increases vascular resistance and may induce hypertension by inhibiting the Na+ pump. The effects of 0.18 and 18 microg/kg, and 1.8 mg/kg ouabain pretreatment on the phenylephrine (PHE; 0.1, 0.25 and 0.5 microg, in bolus)-evoked pressor responses were investigated using anesthetized normotensive (control and uninephrectomized) and hypertensive (1K1C and DOCA-salt treated) rats. Treatment with 18 microg/kg ouabain increased systolic and diastolic blood pressure in all groups studied. However, the magnitude of this increase was larger for the hypertensive 1K1C and DOCA-salt rats than for normotensive animals, while the pressor effect of 0.18 microg/kg ouabain was greater only in DOCA-salt rats. A very large dose (1.8 mg/kg) produced toxic effects on the normotensive control but not on uninephrectomized or 1K1C rats. Rat tail vascular beds were perfused to analyze the effects of 10 nM ouabain on the pressor response to PHE. In all animals, 10 nM ouabain increased the PHE pressor response, but this increase was larger in hypertensive DOCA-salt rats than in normotensive and 1K1C rats. Results suggested that a) increases in diastolic blood pressure induced by 18 microg/kg ouabain were larger in hypertensive than normotensive rats; b) in DOCA-salt rats, smaller ouabain doses had a stronger effect than in other groups; c) hypertensive and uninephrectomized rats were less sensitive to toxic doses of ouabain, and d) after treatment with 10 nM ouabain isolated tail vascular beds from DOCA-salt rats were more sensitive to the pressor effect of PHE than those from normotensive and 1K1C hypertensive rats. These data suggest that very small doses of ouabain, which might produce nanomolar plasma concentrations, enhance pressor reactivity in DOCA-salt hypertensive rats, supporting the idea that endogenous ouabain may contribute to the increase and maintenance of vascular tone in hypertension.

Transmembrane arrangement of mitochondrial porin or voltage-dependent anion channel (VDAC).

Porin or voltage-dependent anion-selective channel (VDAC) is the main protein responsible for the high permeability of the outer mitochondrial membrane. The mitochondrial porin is mainly composed of sided beta-strands, in analogy with bacterial porin, whose structure has been resolved at 1.8 A resolution. In mitochondrial porins the N-terminal region forms an amphipathic alpha-helix, a structure conserved in organisms very distant from the evolutionary point of view. This part of the protein is exposed to the water phase, as demonstrated by ELISA assays. Various extramembranous loops have been identified by specific proteolytic cleavages. These overall, combined results were used to draw a model of the transmembrane arrangement of mammalian porin. This model is compared to other mitochondrial and bacterial porin models.