Rapid detection of bacteria, Enterococcus faecalis, in airborne particles of Hermosillo, Sonora, México

Aim: The microorganisms present in the air have great biological and economic importance because they can seriously affect living organisms. The aim of this work was to implement a real-time PCR technique as a rapid method for detecting Enterococcus faecalis in filters containing airborne particles, seeking to avoid the loss of time in bacterial culture on agar plates. Methodology: Air filter samples were collected from four monitoring stations. Filter samples with PM10 and TSP particles were subjected to nucleic acid extraction and PCR for identification of E. faecalis. Results: The PCR technique developed in this work showed high sensitivity and good specificity for detecting the presence of E. faecalis in airborne particles. The results of the microbiological analysis using traditional identification techniques confirmed the presence of E. faecalis in all sampling sites. Winter was the period with the highest percentage of positive samples. Interpretation: The results suggest that PCR technique can be used for rapid detection of E. faecalis in filters containing PM10 and TSP particles.

Comportamento ingestivo de bovinos leiteiros alimentados com farelo de crambe / Ingestive behavior of dairy catle fed crambe bran

O objetivo deste trabalho foi estudar os efeitos da utilização de farelo de crambe em substituição ao farelo de soja sobre o comportamento ingestivo de bovinos leiteiros. Foram utilizados quatro machos castrados Holandês x Zebu, fistulados no rúmen, com peso vivo médio de 664kg, distribuídos em delineamento quadrado latino 4 x 4. Os tratamentos consistiram em quatro dietas isoproteicas e isoenergéticas, formuladas com relação volumoso:concentrado 60:40 com base na matéria seca (MS). O volumoso foi composto de silagem de milho (51% MS) e feno de Tifton (49% MS), e o concentrado formulado com níveis crescentes de substituição do farelo de soja pelo farelo de crambe em 0%, 2,8%, 6,4% e 11,0% na MS da dieta. O comportamento ingestivo foi avaliado por meio do método direto de avaliação visual, em intervalos de 10 minutos, durante períodos de 24 horas. Registraram-se a frequência de alimentação, a ruminação e o ócio, bem como a posição do animal (em pé ou em decúbito). As variáveis em pé e em decúbito não diferiram entre os tratamentos, assim como os tempos gastos em alimentação, ruminação e ócio. O consumo de matéria seca e de FDN expressos em g/dia e gFDN/dia, respectivamente, a eficiência de ruminação expressa em gMS/min, a eficiência de ruminação expressa em gFDN/dia e o tempo de mastigação total não diferiram significativamente. No entanto, a eficiência de alimentação (gMS/min) variou de forma linear decrescente com a inclusão do farelo de crambe. Os períodos do dia influenciaram todas as atividades. O maior tempo de alimentação foi observado nos períodos após o fornecimento da dieta, e a maior atividade de ruminação foi verificada no período noturno. A substituição de farelo de soja por farelo de crambe não afetou o comportamento ingestivo, exceto para o parâmetro eficiência de alimentação. Nesse sentido, considerando o comportamento ingestivo, recomenda-se a substituição do farelo de soja por farelo de crambe para alimentação de bovinos leiteiros.

The aim of this work was to study the effects of the use of crambe meal replacing soy bean meal on the ingestive behavior of dairy cattle. Four male Holstein x Zebu rumen, with average live weight of 664 kg distributed in a 4x4 latin square design. The treatments consisted of four isonitrogenous and isocaloric diets, formulated with a roughage:concentrate ratio of 60:40 based on dry matter (DM). The forage was corn silage (51% DM) and Tifton grass hay (49% DM), and the concentrate was formulated with increasing levels of substitution of soy bean meal by crambe meal at 0%, 2.8%, 6.4% and 11.0% of the diet DM. The feeding behavior was evaluated using the direct method of visual assessment in 10 minute intervals during a 24 hour period. The frequency of feeding, idling time and the animal's position (standing or supine) were recorded. The variables standing and supine did not differ between treatments as well as time spent feeding, ruminating and idling times. The consumption of dry matter and NDF expressed in g/day and gNDF/day respectively, the efficiency of rumination expressed in gDM/min, the efficiency of rumination expressed in gNDF/day and the total chewing time did not differ significantly. However, the efficiency of power (gDM/min) decreased linearly with the inclusion of crambe meal. Periods of the day influenced all the activities. The longer feeding time was observed in the periods after the diet intake and greater rumination activity was observed at night. The replacement of soy bean meal by meal crambe did not affect feeding behavior, except for the feeding efficiency parameter. In this sense, considering the feeding behavior, the replacement of soy bean meal by crambe meal to feed dairy cattle is recommended.

Basic life support knowledge of first-year university students from Brazil

We aimed to evaluate knowledge of first aid among new undergraduates and whether it is affected by their chosen course. A questionnaire was developed to assess knowledge of how to activate the Mobile Emergency Attendance Service - MEAS (Serviço de Atendimento Móvel de Urgência; SAMU), recognize a pre-hospital emergency situation and the first aid required for cardiac arrest. The students were also asked about enrolling in a first aid course. Responses were received from 1038 of 1365 (76.04%) new undergraduates. The questionnaires were completed in a 2-week period 1 month after the beginning of classes. Of the 1038 respondents (59.5% studying biological sciences, 11.6% physical sciences, and 28.6% humanities), 58.5% knew how to activate the MEAS/SAMU (54.3% non-biological vs 61.4% biological, P=0.02), with an odds ratio (OR)=1.39 (95%CI=1.07-1.81) regardless of age, sex, origin, having a previous degree or having a relative with cardiac disease. The majority could distinguish emergency from non-emergency situations. When faced with a possible cardiac arrest, 17.7% of the students would perform chest compressions (15.5% non-biological vs 19.1% biological first-year university students, P=0.16) and 65.2% would enroll in a first aid course (51.1% non-biological vs 74.7% biological, P<0.01), with an OR=2.61 (95%CI=1.98-3.44) adjusted for the same confounders. Even though a high percentage of the students recognized emergency situations, a significant proportion did not know the MEAS/SAMU number and only a minority had sufficient basic life support skills to help with cardiac arrest. A significant proportion would not enroll in a first aid course. Biological first-year university students were more prone to enroll in a basic life support course.

Time-course effects of aerobic exercise training on cardiovascular and renal parameters in 2K1C renovascular hypertensive rats

Exercise training (Ex) has been recommended for its beneficial effects in hypertensive states. The present study evaluated the time-course effects of Ex without workload on mean arterial pressure (MAP), reflex bradycardia, cardiac and renal histology, and oxidative stress in two-kidney, one-clip (2K1C) hypertensive rats. Male Fischer rats (10 weeks old; 150–180 g) underwent surgery (2K1C or SHAM) and were subsequently divided into a sedentary (SED) group and Ex group (swimming 1 h/day, 5 days/week for 2, 4, 6, 8, or 10 weeks). Until week 4, Ex decreased MAP, increased reflex bradycardia, prevented concentric hypertrophy, reduced collagen deposition in the myocardium and kidneys, decreased the level of thiobarbituric acid-reactive substances (TBARS) in the left ventricle, and increased the catalase (CAT) activity in the left ventricle and both kidneys. From week 6 to week 10, however, MAP and reflex bradycardia in 2K1C Ex rats became similar to those in 2K1C SED rats. Ex effectively reduced heart rate and prevented collagen deposition in the heart and both kidneys up to week 10, and restored the level of TBARS in the left ventricle and clipped kidney and the CAT activity in both kidneys until week 8. Ex without workload for 10 weeks in 2K1C rats provided distinct beneficial effects. The early effects of Ex on cardiovascular function included reversing MAP and reflex bradycardia. The later effects of Ex included preventing structural alterations in the heart and kidney by decreasing oxidative stress and reducing injuries in these organs during hypertension.

Silagem de capim-elefante adicionada de casca de maracujá / Elephant grass silage added to passion fruit's peel

Avaliaram-se-se as características fermentativas e químico-bromatológicas da silagem de capim-elefante contendo diferentes níveis de inclusão do coproduto da extração da polpa do maracujá, a casca de maracujá (CM). Utilizaram-se níveis crescentes de adição desse coproduto (0,0%; 12,5%; 25,0%; 37,5% e 50,0%) em relação à matéria natural do capim-elefante durante a ensilagem. As silagens foram obtidas a partir do corte da forrageira aos 150 dias de idade. O material foi ensilado em silos laboratoriais e, após 180 dias, os silos foram abertos e coletaram-se amostras para a determinação dos teores de matéria seca (MS); carboidratos solúveis (CHOS); potencial hidrogeniônico (pH); nitrogênio amoniacal em porcentagem do nitrogênio total (NNH3/NT); extrato etéreo (EE); proteína bruta (PB); proteína insolúvel em detergente neutro em porcentagem do nitrogênio total (PIDN/NT); proteína insolúvel em detergente ácido em porcentagem do nitrogênio total (PIDA/NT); fibra em detergente neutro (FDN); fibra em detergente ácido (FDA); celulose (CEL); lignina (LIG); nitrogênio insolúvel em detergente neutro (NIDN); nitrogênio insolúvel em detergente ácido (NIDA) e digestibilidade in vitro da matéria seca (DIVMS). Para a avaliação do efeito dos tratamentos, utilizou-se delineamento experimental inteiramente ao acaso, com quatro repetições, executando-se o estudo de regressão para cada variável analisada (P<0,05). De acordo com as equações de regressão, para as características fermentativas, as silagens podem ser consideradas de muito boa qualidade. Quanto às frações fibrosas da silagem, a inclusão de níveis crescentes desse coproduto contribuiu para diminuí-las, o que resultou em maior DIVMS. Os parâmetros analisados permitem concluir que a inclusão da CM in natura durante a ensilagem do capim-elefante é recomendada em todos os níveis avaliados.

We evaluated the fermentative, chemical and qualitative characteristics of elephant grass silage containing different levels of inclusion of coproduct extraction of the passion fruit pulp in nature, passion fruit peel (CM). We used increasing levels of addition of this coproduct (0.0, 12.5, 25.0, 37.5 and 50.0%) in relation to natural field of elephant grass during ensiling. The silages were obtained from the cut forage at 150 days of age. The material was ensiled in laboratory silos, and after 180 days, the silos were opened and samples were collected for determination of dry matter (DM); soluble carbohydrates (CHOS); hydrogen potential (pH); ammonia nitrogen in percentage of total nitrogen (NNH3/NT); ether extract (EE); crude protein (CP); neutral detergent insoluble protein as a percentage of total nitrogen (NDIP/NT); acid detergent insoluble protein as a percentage of total nitrogen (PIDA/NT); neutral detergent fiber (NDF); acid detergent fiber (ADF); cellulose (CEL); lignin (LIG); neutral detergent insoluble nitrogen (NDIN); acid detergent insoluble nitrogen (ADIN) and in vitro digestibility of dry matter (IVDDM). To assess the effect of the treatments we used a completely randomized design with four replications, running the regression analysis for each variable analyzed (P<0.05). According to the regression equations for the fermentative characteristics, silage can be considered very good quality. As for fiber digestibility of silage, the increasing levels of this coproduct contributed to decrease it, which resulted in higher IVDDM. The parameters analyzed showed that the inclusion of CM in nature with elephant grass silage is recommended at all levels evaluated.

Effects of short-term administration of estradiol on reperfusion arrhythmias in rats of different ages

Little is known about age-related differences in short-term effects of estradiol on ischemia-reperfusion (I/R) insults. The present study was designed to evaluate the effects of short-term treatment with estradiol on reperfusion arrhythmias in isolated hearts of 6-7-week-old and 12-14-month-old female rats. Wistar rats were sham-operated, ovariectomized and treated with vehicle or ovariectomized and treated with 17β-estradiol (E2; 5 µg·100 g-1·day-1) for 4 days. Hearts were perfused by the Langendorff technique. Reperfusion arrhythmias, i.e., ventricular tachycardia and/or ventricular fibrillation, were induced by 15 min of left coronary artery ligation and 30 min of reperfusion. The duration and incidence of I/R arrhythmias were significantly higher in young rats compared to middle-aged rats (arrhythmia severity index: 9.4 ± 1.0 vs 3.0 ± 0.3 arbitrary units, respectively, P < 0.05). In addition, middle-aged rats showed lower heart rate, systolic tension and coronary flow. Four-day E2 treatment caused an increase in uterine weight. Although E2 administration had no significant effect on the duration of I/R arrhythmias in middle-aged rats, it induced a marked reduction in the rhythm disturbances of young rats accompanied by a decrease in heart rate of isolated hearts. Also, this reduction was associated with an increase in QT interval. No significant changes were observed in the QT interval of middle-aged E2-treated rats. These data demonstrate that short-term estradiol treatment protects against I/R arrhythmias in hearts of young female rats. The anti-arrhythmogenic effect of estradiol might be related to a lengthening of the QT interval.

Cardiac and renal effects induced by different exercise workloads in renovascular hypertensive rats

We examined the effect of exercise training (Ex) without (Ex 0 percent) or with a 3 percent workload (Ex 3 percent) on different cardiac and renal parameters in renovascular hypertensive (2K1C) male Fisher rats weighing 150-200 g. Ex was performed for 5 weeks, 1 h/day, 5 days/week. Ex 0 percent or Ex 3 percent induced similar attenuation of baseline mean arterial pressure (MAP, 119 ± 5 mmHg in 2K1C Ex 0 percent, N = 6, and 118 ± 5 mmHg in 2K1C Ex 3 percent, N = 11, vs 99 ± 4 mmHg in sham sedentary (Sham Sed) controls, N = 10) and heart rate (HR, bpm) (383 ± 13 in 2K1C Ex 0 percent, N = 6, and 390 ± 14 in 2K1C Ex 3 percent, N = 11 vs 371 ± 11 in Sham Sed, N = 10,). Ex 0 percent, but not Ex 3 percent, improved baroreflex bradycardia (0.26 ± 0.06 ms/mmHg, N = 6, vs 0.09 ± 0.03 ms/mmHg in 2K1C Sed, N = 11). Morphometric evaluation suggested concentric left ventricle hypertrophy in sedentary 2K1C rats. Ex 0 percent prevented concentric cardiac hypertrophy, increased cardiomyocyte diameter and decreased cardiac vasculature thickness in 2K1C rats. In contrast, in 2K1C, Ex 3 percent reduced the concentric remodeling and prevented the increase in cardiac vasculature wall thickness, decreased the cardiomyocyte diameter and increased collagen deposition. Renal morphometric analysis showed that Ex 3 percent induced an increase in vasculature wall thickness and collagen deposition in the left kidney of 2K1C rats. These data suggest that Ex 0 percent has more beneficial effects than Ex 3 percent in renovascular hypertensive rats.

Adaptation of the Medical Research Council Dyspnea Scale and the Oxygen Cost Diagram for its use in Puerto Rico

INTRODUCTION: Dyspnea is a common and disabling symptom for patients with chronic lung diseases. The Medical Research Council Dyspnea Scale (MRC) and the Oxygen Cost Diagram (OCD) are widely used instruments validated in English to measure breathlessness. OBJECTIVE: To translate and culturally adapt the MRC and the OCD for its use in Puerto Rico. METHOD: The scales were translated to Spanish and back translated. They were tested in patients attending a pulmonary clinic to assess its relevance and comprehension. Subjects answered the instruments, had a structured clinical interview and provided feedback. A multidisciplinary committee analyzed the source of misunderstanding using the input of the subjects, the clinical physicians, and the evaluators; made adjustments, and retested the instruments until inconsistencies were not observed. RESULTS: Placing emphasis on time spent walking instead of distance traveled improved the discrimination between grade two and three in the MRC. In the OCD, placing the activities to the right side eased the interpretation of the scale. Numerical symbols were eliminated to minimize discomfort in the severely impaired subjects reluctant to mark the line near zero. Reversing the order of both scales encouraged a thorough reading of the activities from minimal to high energy demanding eliciting a more fitting response compared to structured clinical interview especially in the severely ill patient. CONCLUSION: Using cross-cultural research methodologies to translate the MRC and OCD allowed the identification of differences in conceptualization when assessing the severity of dyspnea in Puerto Rico. Further testing is needed to confirm psychometric properties.

Reduced plasma levels of angiotensin-(1-7) and renin activity in preeclamptic patients are associated with the angiotensin I- converting enzyme deletion/deletion genotype

The relationship between preeclampsia and the renin-angiotensin system (RAS) is poorly understood. Angiotensin I-converting enzyme (ACE) is a key RAS component and plays an important role in blood pressure homeostasis by generating angiotensin II (Ang II) and inactivating the vasodilator angiotensin-(1-7) (Ang-(1-7)). ACE (I/D) polymorphism is characterized by the insertion (I) or deletion (D) of a 287-bp fragment, leading to changes in ACE activity. In the present study, ACE (I/D) polymorphism was correlated with plasma Ang-(1-7) levels and several RAS components in both preeclamptic (N = 20) and normotensive pregnant women (N = 20). The percentage of the ACE DD genotype (60 percent) in the preeclamptic group was higher than that for the control group (35 percent); however, this percentage was not statistically significant (Fisher exact test = 2.86, d.f. = 2, P = 0.260). The highest plasma ACE activity was observed in the ACE DD preeclamptic women (58.1 ± 5.06 vs 27.6 ± 3.25 nmol Hip-His Leu-1 min-1 mL-1 in DD control patients; P = 0.0005). Plasma renin activity was markedly reduced in preeclampsia (0.81 ± 0.2 vs 3.43 ± 0.8 ng Ang I mL plasma-1 h-1 in DD normotensive patients; P = 0.0012). A reduced plasma level of Ang-(1-7) was also observed in preeclamptic women (15.6 ± 1.3 vs 22.7 ± 2.5 pg/mL in the DD control group; P = 0.0146). In contrast, plasma Ang II levels were unchanged in preeclamptic patients. The selective changes in the RAS described in the present study suggest that the ACE DD genotype may be used as a marker for susceptibility to preeclampsia.

Cardiovascular actions of angiotensin-(1-7)

Angiotensin-(1-7) (Ang-(1-7)) is now considered to be a biologically active member of the renin-angiotensin system. The functions of Ang-(1-7) are often opposite to those attributed to the main effector component of the renin-angiotensin system, Ang II. Chronic administration of angiotensin-converting enzyme inhibitors (ACEI) increases 10- to 25-fold the plasma levels of this peptide, suggesting that part of the beneficial effects of ACEI could be mediated by Ang-(1-7). Ang-(1-7) can be formed from Ang II or directly from Ang I. Other enzymatic pathways for Ang-(1-7) generation have been recently described involving the novel ACE homologue ACE2. This enzyme can form Ang-(1-7) from Ang II or less efficiently by the hydrolysis of Ang I to Ang-(1-9) with subsequent Ang-(1-7) formation. The biological relevance of Ang-(1-7) has been recently reinforced by the identification of its receptor, the G-protein-coupled receptor Mas. Heart and blood vessels are important targets for the formation and actions of Ang-(1-7). In this review we will discuss recent findings concerning the biological role of Ang-(1-7) in the heart and blood vessels, taking into account aspects related to its formation and effects on these tissues. In addition, we will discuss the potential of Ang-(1-7) and its receptor as a target for the development of new cardiovascular drugs.

Genotoxic effects of rotenone on cultured lymphocytes

Rotenone is a heterocyclic compound widely used as an insecticide, acaricide and piscicide. Its toxicity is mainly caused by the inhibition of mitochondrial respiratory processes and ATP production, resulting in the generation of reactive oxygen species. Reactive oxygen species can interact with DNA, RNA and proteins, leading to cell damage, followed by death. We used the Comet assay, and we analyzed chromosome aberrations, in order to evaluate the genotoxic and clastogenic effects of rotenone on the different phases of the cell cycle. Cultured human lymphocytes were treated with 1.0, 1.5 and 2.0 microg/mL rotenone during the G1, G1/S, S (pulses of 1 and 6 h), and G2 phases of the cell cycle. Rotenone induced DNA damage and was clastogenic, but the clastogenicity was detected only with treatments conducted during the G1/S and S phases of the cell cycle. Rotenone also induced endoreduplication and polyploidy in treatments made during G1, while it significantly reduced the mitotic index in all phases of the cell cycle.

Angiotensin-(1-7) improves the post-ischemic function in isolated perfused rat hearts

We evaluated the effects of angiotensin-(1-7) (Ang-(1-7)) on post-ischemic function in isolated hearts from adult male Wistar rats perfused according to the Langendorff technique. Local ischemia was induced by coronary ligation for 15 min. After ischemia, hearts were reperfused for 30 min. Addition of angiotensin II (Ang II) (0.20 nM, N = 10) or Ang-(1-7) (0.22 nM, N = 10) to the Krebs-Ringer perfusion solution (KRS) before the occlusion did not modify diastolic or systolic tension, heart rate or coronary flow (basal values for Ang-(1-7)-treated hearts: 0.72 ± 0.08 g, 10.50 ± 0.66 g, 216 ± 9 bpm, 5.78 ± 0.60 ml/min, respectively). During the period of occlusion, the coronary flow, heart rate and systolic tension decreased (values for Ang-(1-7)-treated hearts: 2.83 ± 0.24 ml/min, 186 ± 7 bpm, 6.95 ± 0.45 g, respectively). During reperfusion a further decrease in systolic tension was observed in control (4.95 ± 0.60 g) and Ang II-treated hearts (4.35 ± 0.62 g). However, in isolated hearts perfused with KRS containing Ang-(1-7) the further reduction of systolic tension during the reperfusion period was prevented (7.37 ± 0.68 g). The effect of Ang-(1-7) on the systolic tension was blocked by the selective Ang-(1-7) antagonist A-779 (2 nM, N = 9), by the bradykinin B2 antagonist HOE 140 (100 nM, N = 10), and by indomethacin pretreatment (5 mg/kg, ip, N = 8). Pretreatment with L-NAME (30 mg/kg, ip, N = 8) did not change the effect of Ang-(1-7) on systolic tension (6.85 ± 0.61 g). These results show that Ang-(1-7) at low concentration (0.22 nM) improves myocardial function (systolic tension) in ischemia/reperfusion through a receptor-mediated mechanism involving release of bradykinin and prostaglandins

Standardization of a fluorimetric assay for the determination of tissue angiotensin-converting enzyme activity in rats

The tripeptide Hip-His-Leu was used to standardize a fluorimetric method to measure tissue angiotensin-converting enzyme (ACE) activity in rats. The fluorescence of the o-phthaldialdehyde-His-Leu adduct was compared in the presence and absence of the homogenate (25 µl) to determine whether the homogenate from different tissues interfered with the fluorimetric determination of the His-Leu product. Only homogenates from lung and renal medulla and cortex showed significantly altered fluorescence intensity. To overcome this problem, the homogenate from these tissues were diluted 10 times with assay buffer. The specificity of the assay was demonstrated by the inhibition of ACE activity with 3 µM enalaprilat (MK-422). There was a linear relationship between product formation and incubation time for up to 90 min for homogenates of renal cortex and medulla and liver, for up to 60 min for ventricles and adrenals and for up to 30 min for the aorta, lung and atrium homogenates. In addition, there was a linear relationship between product formation and the amount of protein in the homogenates within the following range: lung, 30-600 µg; renal cortex and medulla, 40-400 µg; atrium and ventricles, 20-200 µg; adrenal, 20-100 µg; aorta, 5-100 µg; liver, 5-25 µg. No peptidase activity against the His-Leu product (31 nmol), assayed in borate buffer (BB), was detected in the different homogenates except the liver homogenate, which was inhibited by 0.1 mM r-chloromercuribenzoic acid. ACE activity in BB was higher than in phosphate buffer (PB) due, at least in part, to a greater hydrolysis of the His-Leu product in PB. ACE activity of lung increased 20 percent when BB plus Triton was used. Enzyme activity was stable when the homogenates were stored at -20o or -70oC for at least 30 days. These results indicate a condition whereby ACE activity can be easily and efficiently assayed in rat tissue samples homogenized in BB using a fluorimetric method with Hip-His-Leu as a substrate.

Angiotensin-(1-7) potentiates the coronary vasodilatatory effect of bradykinin in the isolated rat heart

It has been shown that angiotensin-(1-7) (Ang-(1-7)) infusion potentiates the bradykinin (BK)-induced hypotensive response in conscious rats. The present study was conducted to identify Ang-(1-7)-BK interactions in the isolated rat heart perfused according to the Langendorff technique. Hearts were excised and perfused through the aortic stump under a constant flow with Krebs-Ringer solution and the changes in perfusion pressure and heart contractile force were recorded. Bolus injections of BK (2.5, 5, 10 and 20 ng) produced a dose-dependent hypotensive effect. Ang-(1-7) added to the perfusion solution (2 ng/ml) did not change the perfusion pressure or the contractile force but doubled the hypotensive effect of the lower doses of BK. The BK-potentiating Ang-(1-7) activity was blocked by pretreatment with indomethacin (5 mg/kg, ip) or L-NAME (30 mg/kg, ip). The Ang-(1-7) antagonist A-779 (50 ng/ml in Krebs-Ringer) completely blocked the effect of Ang-(1-7) on BK-induced vasodilation. These data suggest that the potentiation of the BK-induced vasodilation by Ang-(1-7) can be attributed to the release of nitric oxide and vasodilator prostaglandins through an Ang-(1-7) receptor-mediated mechanism.

Effect of selective angiotensin antagonists on the antidiuresis produced by angiotensin-(1-7) in water-loaded rats

In the present study we evaluated the nature of angiotensin receptors involved in the antidiuretic effect of angiotensin-(1-7) (Ang-(1-7)) in water-loaded rats. Water diuresis was induced in male Wistar rats weighing 280 to 320 g by water load (5 ml/100 g body weight by gavage). Immediately after water load the rats were treated subcutaneously with (doses are per 100 g body weight): 1) vehicle (0.05 ml 0.9 percenr NaCl); 2) graded doses of 20, 40 or 80 pmol Ang-(1-7); 3) 200 nmol Losartan; 4) 200 nmol Losartan combined with 40 pmol Ang-(1-7); 5) 1.1 or 4.4 nmol A-779; 6) 1.1 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang-(1-7); 7) 4.4 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang-(1-7); 8) 95 nmol CGP 42112A, or 9) 95 nmol CGP 42112A combined with 40 pmol Ang-(1-7). The antidiuretic effect of Ang-(1-7) was associated with an increase in urinary Na+ concentration, an increase in urinary osmolality and a reduction in creatinine clearance (CCr: 0.65 ñ 0.04 ml/min vs 1.45 ñ 0.18 ml/min in vehicle-treated rats, P<0.05). A-779 and Losartan completely blocked the effect of Ang-(1-7) on water diuresis (2.93 ñ 0.34 ml/60 min and 3.39 ñ 0.58 ml/60 min, respectively). CGP 42112A, at the dose used, did not modify the antidiuretic effect of Ang-(1-7). The blockade produced by Losartan was associated with an increase in CCr and with an increase in sodium and water excretion as compared with Ang-(1-7)-treated rats. When Ang-(1-7) was combined with A-779 there was an increase in CCr and natriuresis and a reduction in urine osmolality compared with rats treated with Ang-(1-7) alone. The observation that both A-779, which does not bind to AT1 receptors, and Losartan blocked the effect of Ang-(1-7) suggests that the kidney effects of Ang-(1-7) are mediated by a non-AT1 angiotensin receptor that is recognized by Losartan.

A comparative analysis of preprocessing techniques of cardiac event series for the study of heart rhythm variability using simulated signals

In the present study, using noise-free simulated signals, we performed a comparative examination of several preprocessing techniques that are used to transform the cardiac event series in a regularly sampled time series, appropriate for spectral analysis of heart rhythm variability (HRV). First, a group of noise-free simulated point event series, which represents a time series of heartbeats, was generated by an integral pulse frequency modulation model. In order to evaluate the performance of the preprocessing methods, the differences between the spectra of the preprocessed simulated signals and the true spectrum (spectrum of the model input modulating signals) were surveyed by visual analysis and by contrasting merit indices. It is desired that estimated spectra match the true spectrum as close as possible, showing a minimum of harmonic components and other artifacts. The merit indices proposed to quantify these mismatches were the leakage rate, defined as a measure of leakage components (located outside some narrow windows centered at frequencies of model input modulating signals) with respect to the whole spectral components, and the numbers of leakage components with amplitudes greater than 1 percent, 5 percent and 10 percent of the total spectral components. Our data, obtained from a noise-free simulation, indicate that the utilization of heart rate values instead of heart period values in the derivation of signals representative of heart rhythm results in more accurate spectra. Furthermore, our data support the efficiency of the widely used preprocessing technique based on the convolution of inverse interval function values with a rectangular window, and suggest the preprocessing technique based on a cubic polynomial interpolation of inverse interval function values and succeeding spectral analysis as another efficient and fast method for the analysis of HRV signals.

Effect of angiotensin-(1-7) on reperfusion arrhythmias in isolated rat hearts

There is increasing evidence that angiotensin-(1-7) (Ang-(1-7) is an endogenous biologically active component of the renin-angiotensin system (RAS). In the present study, we investigated the effects of Ang-(1-7) on reperfusion arrhythmias in isolated rat hearts. Isolated rat hearts were perfused with two different media, i.e., Krebs-Ringer (2.52 mM CaCl2) and low-Ca2+ Krebs-Ringer (1.12 mM Cacl2) and low-Ca2+ Krebs-Ringer (1.12 mM CaCl2). In hearts perfused with Krebs-Ringer, Ang-(1-7) produced a concentration-dependent (27-210 nM) reduction in coronary flow (25 percent reduction at highest concentration), while only slight and variable changes in contaction force and heart rate were observed. Under the same conditions, angiotensin II (Ang II; 27 and 70 nM) produced a significant reduction in coronary flow (39 percent and 48 percent, respectively) associated with a significant increase in force. A decrease in heart rate was also observed. In low-Ca2+ Krebs-Ringer solution, perfusion with Ang-(1-7) or Ang II at 27 nM concentration produced similar changes in coronary flow, contraction force and heart rate. In isolated hearts perfused with normal Krebs- Ringer, Ang-(1-7) produced a significant enhancement of reperfusion arrhythmias revealed by an increase in the incidence and duration of ventricular tachycardia and ventricular fibrillation (more than 30-min duration). The faciliation of reperfusion arrhythmias by Ang-(1-7) was associated with an increase in the magnitude of the decreased force usually observed during the postischemic period. The effects of Ang-(1-7) were abolished in isolated rat hearts perfused with low-Ca2+ Krebs-Ringer. The effect of Ang II (27 nM) was similar but less pronounced than that of Ang-(1-7) at the same concentration. These results indicate that the heart is a site of action for Ang-(1-7) and suggest that this heptapeptide may be involved in the mediation of the cardiac effects of the RAS.

Renal actions of angiotensin-(1-7)

The heptapeptide angiotensin-(1-7) is considered to be a biologically active endproduct of the renin-angiotensin system. This angiotensin, which is devoid of the most known actions of angioatensin II such as induction of drinking behavior and vasoconstriction, has several selective effects in the brain and periphery. In the present article we briefly review recent evidence for a physiological role of angiotensin-(1-7) in the control of hydroelectrolyte balance.

Central and peripheral actions of angiotensin-(1-7)

1. In this article we review the physiological actions of the heptapeptide angiotensin-(1-7) [Ang-(1-7)] at the periphery and on central pathways involved in the control of arterial pressure. Peripherally Ang-(1-7) has been shown to present a potent antidiuretic effect on water-loaded rats. Microinjection of pmol amounts of Ang-(1-7) into the dorsomedial or ventrolateral medulla (VLM) of anesthetized rats produces cardiovascular effects comparable to Ang II. In addition, in vitro experiments have shown that Ang-(1-7) has a potent vasopressin and prostaglandin releasing activity and excites neuronal activity in the hypothalamus and medulla. 2. Evidence for the existence of a new angiotensin receptor subtype that mediates the central cardiovascular actions of this active peptide of the renin-angiotensin system (RAS) is also provided. Neither the AT1 receptor antagonist DUP 753 or the AT2 receptor antagonist CGP 42112A blocked the pressor response produced by microinjection of Ang-(1-7) into the rostral VLM. However, the effect of Ang-(1-7) on VLM was completely abolished by the non-specific angiotensin receptor antagonist, Sar1-Thr8-Ang II. 3. The data presented here reinforce the hypothesis of the existence of complex site-specific interactions between multiple angiotensins and multiple receptors in the mediation of important central and peripheral effects of the RAS.

Angiotensin-1(1-7) is a potent antidiuretic peptide in rats

We have shown previously that angiotensin (1-7) (Asp-Arg Val-Tyr-Ile-His-Pro) is a biologically active endogenous angiotensin which is a major product of angiotensin I processing by an angiotensin converting enzyme (ACE)-independent pathway. Intense staining for angiotensin (1-7) immunoreactivity was demonstrable in brain areas related to the maintenance of hydromineral balance, suggesting the involvement of this peptide in this process. In the present study we investigated the antidiuretic effect of angiotensin (1-7), by determining its effect on the water diuresis produced by an ip water load (5 ml/100 g) in male Wistar rats. The peptide had a pronounced antidiuretic effect when administered peripherally in doses ranging from 5.5 to 22 pmol/100 g. In contrast, angiotensin - (1-7) and vasopressin (AVP) showed that both peptides act in the same molar range althouth AVP was slightly more potent than angiotensin (1-7). Urine volumes for 22 pmol/100 g angiotensin (1-7) were 0.85 ñ 0.26 and 3.47 ñ 0.36 ml for hours 1 and 2, respectively, whereas they were 0.54 ñ 0.40 and 2.38 ñ 0.64 ml for 10 pmol/100 g AVP. There was apparent additivity of effect when 10 pmol of each peptide were administered simultaneously (0.0 and 1.72 ñ 0.45 ml vs 2.58 ñ 0.45 and 3.85 ñ 0.35 ml for hours 1 and 2, respectively). The unexpected potency of this peptide in reducing water diuresis and its localization in brain areas related to AVP release suggest a role for angiotensin (1-7) in the mechanisms involved in the maintenance of hydroelectrolyte balance