Acute effect of passive cycle-ergometry and functional electrical stimulation on nitrosative stress and inflammatory cytokines in mechanically ventilated critically ill patients: a randomized controlled trial.

Early mobilization is beneficial for critically ill patients because it reduces muscle weakness acquired in intensive care units. The objective of this study was to assess the effect of functional electrical stimulation (FES) and passive cycle ergometry (PCE) on the nitrous stress and inflammatory cytometry in critically ill patients. This was a controlled, randomized, open clinical trial carried out in a 16-bed intensive care unit. The patients were randomized into four groups: Control group (n=10), did not undergo any therapeutic intervention during the study; PCE group (n=9), lower-limb PCE for 30 cycles/min for 20 min; FES group (n=9), electrical stimulation of quadriceps muscle for 20 min; and FES with PCE group (n=7), patients underwent PCE and FES, with their order determined randomly. The serum levels of nitric oxide, tumor necrosis factor alpha, interferon gamma, and interleukins 6 and 10 were analyzed before and after the intervention. There were no differences in clinical or demographic characteristics between the groups. The results revealed reduced nitric oxide concentrations one hour after using PCE (P<0.001) and FES (P<0.05), thereby indicating that these therapies may reduce cellular nitrosative stress when applied separately. Tumor necrosis factor alpha levels were reduced after the PCE intervention (P=0.049). PCE and FES reduced nitric oxide levels, demonstrating beneficial effects on the reduction of nitrosative stress. PCE was the only treatment that reduced the tumor necrosis factor alpha concentration.

Acute effect of passive cycle-ergometry and functional electrical stimulation on nitrosative stress and inflammatory cytokines in mechanically ventilated critically ill patients: a randomized controlled trial

Early mobilization is beneficial for critically ill patients because it reduces muscle weakness acquired in intensive care units. The objective of this study was to assess the effect of functional electrical stimulation (FES) and passive cycle ergometry (PCE) on the nitrous stress and inflammatory cytometry in critically ill patients. This was a controlled, randomized, open clinical trial carried out in a 16-bed intensive care unit. The patients were randomized into four groups: Control group (n=10), did not undergo any therapeutic intervention during the study; PCE group (n=9), lower-limb PCE for 30 cycles/min for 20 min; FES group (n=9), electrical stimulation of quadriceps muscle for 20 min; and FES with PCE group (n=7), patients underwent PCE and FES, with their order determined randomly. The serum levels of nitric oxide, tumor necrosis factor alpha, interferon gamma, and interleukins 6 and 10 were analyzed before and after the intervention. There were no differences in clinical or demographic characteristics between the groups. The results revealed reduced nitric oxide concentrations one hour after using PCE (P<0.001) and FES (P<0.05), thereby indicating that these therapies may reduce cellular nitrosative stress when applied separately. Tumor necrosis factor alpha levels were reduced after the PCE intervention (P=0.049). PCE and FES reduced nitric oxide levels, demonstrating beneficial effects on the reduction of nitrosative stress. PCE was the only treatment that reduced the tumor necrosis factor alpha concentration.

Morphological characterization of hemocytes from Biomphalaria glabrata and Biomphalaria straminea.

Biomphalaria glabrata and Biomphalaria straminea have been identified as intermediate hosts for Schistosoma mansoni. Several studies have found two cell types in the hemolymph of B. glabrata (hyalinocytes and granulocytes). However, there are no studies describing the hemocytes of B. straminea. With the aim of further describing the hemocyte subsets in B. glabrata and B. straminea, we conducted a detailed study using optical microscopy and transmission electron microscopy. Based on the morphological characteristics of the cells, we identified the same types of hemocytes in two species of molluscs, namely: blast-like cells, granulocytes, type I hyalinocytes, type II hyalinocytes and type III hyalinocytes. Blast-like cells had a spherical profile with a central nucleus filling almost the whole cell. Granulocytes were characterized by presenting variable numbers of granules. Type I hyalinocytes were the most abundant cell type and displayed various cytoplasmic projections. Type II and type III hyalinocytes had never previously been reported. They were few in number and were characterized by having an eccentric nucleus. From these results, it is concluded that there are five types of cells in the hemolymph of B. glabrata and B. straminea. Further studies are now needed to identify the role of these hemocytes in the immune response of these snails.

Apolipoprotein E genotype as a determinant of survival in chronic lymphocytic leukemia.

Survival of chronic lymphocytic leukemia (CLL) cells requires sustained activation of the antiapoptotic PI-3-K/Akt pathway, and many therapies for CLL cause leukemia cell death by triggering apoptosis. Blood lipoprotein particles are either pro- or antiapoptotic. High-density lipoprotein particles are antiapoptotic through sphingosine-1-phosphate receptor 3-mediated activation of the PI-3-K/Akt pathway. Apolipoprotein E4 (apoE4)-very low density lipoproteins (VLDL) increase apoptosis, but the apoE2-VLDL and apoE3-VLDL isoforms do not. As increased B-cell apoptosis favors longer survival of CLL patients, we hypothesized that APOE4 genotype would beneficially influence the clinical course of CLL. We report here that women (but not men) with an APOE4 genotype had markedly longer survival than non-APOE4 patients. VLDL is metabolized to low-density lipoprotein through lipoprotein lipase. Higher levels of lipoprotein lipase mRNA in these CLL patients correlated with shorter survival. The beneficial effect of APOE4 in CLL survival is likely mediated through APOE4 allele-specific regulation of leukemia cell apoptosis. The APOE allele and genotype distribution in these CLL patients is the same as in unaffected control populations, suggesting that although APOE genotype influences CLL outcome and response to therapy, it does not alter susceptibility to developing this disease.

Thie expression of an intraspecific aggressive reaction in the face of a stressor agent alters the immune response in rats.

The repercussion on the immune response of the expression of intraspecific aggressiveness in the face of a stressor agent was investigated in rats. Ninety-day-old animals were divided into three groups: the control group (only immunological measurements were performed), the foot-shock (FS) (animals individually receiving FS), and the intraspecific aggressive response (IAR) group (animals receiving FS and presenting IAR). For immunological measurements, blood samples were collected promptly at 7 and 15 days after FS or IAR. The FS reduced the total leukocyte amount presented. However, aggressiveness triggered not only reduction of the leukocytes, but also lymphocyte decrease and neutrophil increase. Moreover, an elevation in total leukocytes associated with an increase in the humoral immune response was also observed one week after IAR. In this study, the expression of intraspecific aggressiveness in the face of a stressor seemed to activate the immune system and to potentiate the antigen specific humoral response.

The expression of an intraspecific aggressive reaction in the face of a stressor agent alters the immune response in rats

A repercussão sobre a resposta imune da expressão da agressividade intra-específica diante de um estressor foi investigada em ratos. Aos 90 dias de vida, os animais foram divididos em três grupos: grupo-controle (foram realizadas apenas mensurações imunológicas), choque nas patas (FS) (os animais receberam FS individualmente) e grupo resposta agressiva intra-específica (IAR) (os animais receberam FS e apresentaram IAR). Para as medições imunológicas, amostras de sangue foram coletadas imediatamente, 7 e 15 dias após FS ou IAR. O FS reduziu a quantidade total de leucócitos. Contudo, a agressividade foi acompanhada, além da redução do número de leucócitos, por diminuição de linfócitos e aumento de neutrófilos. Além disso, também foi observada elevação no número de leucócitos associada a aumento na resposta imune humoral uma semana após as IAR. Neste estudo, a expressão da agressividade intra-específica diante de um estressor parece ativar o sistema imune e potencializar a resposta humoral antígeno específica.

Malnutrition during brain growth spurt alters the effect of fluoxetine on aggressive behavior in adult rats.

Malnutrition effect during the suckling period on aggressive behavior was investigated in adult rats treated and not treated with fluoxetine, a selective serotonin reuptake inhibitor. Sixty-four Wistar male rats were allocated in two groups, according to their mothers' diet during lactation. The well-nourished group was fed by mothers receiving a 23% protein diet; the malnourished one by mothers receiving a 8% protein diet. Following weaning, all rats received the 23% protein diet. On the 90th day after birth, each nutritional group was divided into two subgroups, one receiving a single daily injection of fluoxetine (10 mg/kg) and the other of a saline solution (0.9% NaCl) for 14 days. Treatment with Fluoxetine reduced aggressive response in well-nourished but not in malnourished rats. These findings suggest that the serotoninergic system was affected by malnutrition during the critical period of brain development, and persisted even after a long period of nutritional recovery.

Early malnourished rats are not affected by anorexia induced by a selective serotonin reuptake inhibitor in adult life.

The effect of early postnatal malnutrition upon food intake and its modulation by the selective serotonin reuptake inhibitor (SSRI) citalopram, was investigated in adult rats. Sixty four Wistar rats were allocated to two groups, according to their mother's diet during lactation. Mothers receiving a 23% protein diet fed the well-nourished group; mothers receiving 8% protein diet fed the malnourished. After weaning, all rats received the 23% protein diet ad libitum. On the 120th day after birth, each nutritional group was divided in two subgroups (each one, n = 16) which received a single daily injection of citalopram (10 mg/kg) or saline (0.9% NaCl) for 14 days. Chronic treatment with citalopram decreased both the food intake and weight gain in the well-nourished rats, but not in the malnourished ones. These data are consistent with findings concerning the nutritional manipulation of the nervous system during its higher vulnerable phase, suggesting that early malnutrition alters the effect of treatment of SSRI in adult rats, and that malnutrition during the critical period of brain development affects the serotoninergic system.

[Lipid profile among school children in Campinas, Brazil]. / Perfil lipídico em escolares de Campinas, SP, Brasil.

OBJECTIVE: To describe the lipid profile and the prevalence of hypercholesterolemia among schoolchildren aged 7 to 14 years in Campinas, São Paulo State, Brazil. METHODS: Plasma cholesterol levels, fractions, ratios and triglycerides were determined according to age and gender in a total of 1,600 schoolchildren. Hypercholesterolemia was considered borderline for 170 mg/dl/=200 mg/dl. RESULTS: Schoolchildren presented a cholesterol mean of 160 mg/dl, HDL-cholesterol mean of 49 mg/dl, LDL-cholesterol mean of 96 mg/dl, VLDL-cholesterol mean of 16 mg/dl, triglycerides mean of 79 mg/dl, cholesterol/HDL-cholesterol mean of 3.5 and LDL-cholesterol/HDL-cholesterol mean of 2.1. In general, females had higher cholesterol and triglycerides values than males. The prevalence of hypercholesterolemia was 35.0%: 15.6% was borderline high, 9.8% moderate and 9.5% severe. Females presented higher prevalence of hypercholesterolemia than males. CONCLUSIONS: The results pointed to the emergence of hypercholesterolemia as a public health problem in Brazil.

Effect of stress on the production of O(2)(-) in alveolar macrophages.

Alveolar macrophages (AM) of male rats (200-250 g), stressed or not, were evaluated with relationship to superoxide production (SP). Plasma levels of corticosterone were measured. The control group showed larger SP than the stressed group in all intervals of time. Exposure in vitro of AM to a synthetic glucocorticoid for 40 min (the same time of restraint stress) inhibits SP. Therefore, it seems under stress situations there is an increase of plasma levels of corticosterone and a decrease of SP in AM after stimulation with PMA. O(2)(-) is a potent microbicide and its reduction could cause the loss of microbicidal activity of AM.

Genomic structure and chromosomal localization of the novel ETS factor, PE-2 (ERF).

The members of the ETS family of transcription factors are grouped because they share a highly conserved DNA binding domain. These factors are involved in growth factor pathways and regulate both proliferation and differentiation. To identify ETS factors that may be involved in early hematopoietic progenitor regulation, we isolated a novel member of the ETS family by reverse transcriptase-PCR of the conserved DNA binding domain using degenerate oligonucleotides. This gene directs the synthesis of a 2704-nucleotide transcript whose largest open reading frame encodes a 548-amino-acid protein. Northern blot analysis reveals ubiquitous expression in all human tissues and cell lines tested, with highest levels in the testis, ovary, pancreas, and heart. Comparison of this gene with the available databases reveals very significant homology to the ETS factor PE-1 and probable near-identity with the recently cloned factor ERF. The PE-2 gene is composed of four exons spanning over 9 kb of genomic DNA. Sequence analysis of the promoter region reveals a GC-rich sequence without a TATA motif and with putative binding motifs for CREB, c-myb, and AP-1 factors. Using mouse-human somatic hybrids and FISH analysis, the PE-2 gene is localized to human chromosome 19q13.2, a region involved in translocations and deletions in leukemias and several solid tumors, suggesting that this novel ETS factor may play a role in carcinogenesis.

5-Lipoxygenase and endotoxin-induced microvascular albumin exchanges and leucocyte recruitment in guinea-pig lungs.

The interference of the 5-lipoxygenase inhibitor, BW B70C ((E)-N-(3-[3-(4-fluorophenoxy)phenyl]-1(R,S)-methyl prop-2-enyl)-N-hydroxyurea), with Escherichia coli lipopolysaccharide (endotoxin)-induced lung leucocyte sequestration and microvascular albumin exchanges was evaluated in the anaesthetised guinea-pig using radioactive tracers, in parallel to the effects on cell counts in the broncho-alveolar lavage fluid, blood tumour necrosis factor (TNF-alpha) content, secretion of phospholipase A2 and synthesis of leukotriene C4 by alveolar macrophages. Intravenous injections of 0.1 or 1 mg/kg endotoxin induced lung leucocyte sequestration but only the higher dose induced an increase in albumin microvascular exchanges and the infiltration of leucocytes towards the airway lumen. Leukotriene B4, a potential mediator of the 5-lipoxygenase-dependent endotoxin effects, induced a rapid and transient lung leucocyte sequestration and leucopenia associated with a more progressive increase in microvascular exchanges. The 5-lipoxygenase inhibitor, BW B70C, injected i.p. (30 mg/kg) prevented leukotriene C4 synthesis by alveolar macrophages and reduced leucocyte migration to the airways lumen as well as albumin microvascular leakage but did not affect the endotoxin-induced increase in the blood level of TNF-alpha and of secreted phospholipase A2. However, BW B70C failed to modify vascular leucocyte margination induced by 1 mg/kg endotoxin, suggesting that, apart from a role of 5-lipoxygenase, alternative pathways operate in response to endotoxin in guinea-pig.

Interference of dexamethasone with leukocyte blood volume and albumin movements in lungs from endotoxemic guinea-pigs.

Using radioactive tracers, we measured blood volume, albumin exchanges and blood leukocyte sequestration within lungs, following an intravenous injection of lipopolysaccharide (0.1-1 mg/kg). Neutrophil infiltration into the airways was followed in parallel experiments. Dexamethasone pretreatment (20 mg/kg, subcutaneous) failed to prevent early pulmonary changes induced by lipopolysaccharide as decreased blood volume, leukocyte sequestration, leukopenia or the increased trans-endothelial albumin exchanges. However, dexamethasone provided a significant protection against the later albumin leakage through the endothelial/epithelial barrier and the neutrophil accumulation in the airways observed in lipopolysaccharide-treated guinea-pigs. Our results indicate that the protective effect of dexamethasone in lipopolysaccharide-induced lung injury might derive from an initial reduction of leukocyte adhesion and a later decrease in alveolo-capillary permeability.

Fenspiride: an anti-inflammatory drug with potential benefits in the treatment of endotoxemia.

Using a model of endotoxemia triggered by the intravenous injection of bacterial lipopolysaccharide (0.1 and 1 mg/kg) to guinea-pigs, we investigated the interference of fenspiride, an anti-inflammatory drug recommended for the treatment of inflammatory diseases of the upper respiratory tract. Administered orally at 60 mg/kg, fenspiride reduced the lipopolysaccharide-induced early rise of tumor necrosis factor concentrations in serum (4.2 +/- 0.9 vs. 2.3 +/- 0.5 ng/ml, P < 0.05) and in the bronchoalveolar lavage fluid (55.7 +/- 20 vs. 19.7 +/- 7.5 ng/ml, P < 0.05). The lipopolysaccharide-induced primed stimulation of alveolar macrophages, defined as their enhanced release of arachidonic acid metabolites as compared to cells from untreated controls upon stimulation with N-formyl-methionyl-phenylalanine was also reduced by fenspiride (1551.5 +/- 183.7 vs. 771.5 +/- 237.5 pg/mu g protein, P < 0.05 for thromboxane B2 and 12.6 +/- 4.9 vs. 3.6 +/- 0.9 pg/ mu g protein, P < 0.05 for leukotriene C4). Finally, fenspiride reduced the increased serum concentrations of extracellular type II phospholipase A2 (3.9 +/- 1.2 vs. 1.2 +/- 0.1 nmol/ml per min, P < 0.01), the intensity of the neutrophilic alveolar invasion and the lethality due to the lipopolysaccharide. The protective effect of fenspiride may result from the inhibition of the formation of tumor necrosis factor, a major mediator of the effects of lipopolysaccharide.

Modulation by dexamethasone of phospholipase A2 activities in endotoxemic guinea pigs.

One hour after lipopolysaccharide (LPS) administration (intravenous) in guinea pigs, alveolar macrophages are primed for an ex vivo increased secretion of arachidonic acid metabolites from the cyclooxygenase and the lipoxygenase pathways, with challenge by a second stimulus. At the same time, maximal levels of tumor necrosis factor-alpha (TNF-alpha) are observed in the circulation and in the bronchoalveolar lavage fluid. An extracellular form of phospholipase A2, corresponding probably to the low-molecular-mass type II enzyme, known to accumulate in inflammatory exudates, appears later in the serum of guinea pigs, to reach maximal levels 6 h after the LPS. Unlike the intracellular enzyme, extracellular phospholipase A2 is not increased by LPS in alveolar macrophages or in bronchoalveolar lavage fluids. After 24 h, at the time when neither TNF-alpha nor extracellular phospholipase A2 is present and priming of macrophages is over, maximal neutrophil infiltration is observed in the alveolar space of LPS-treated guinea pigs. Dexamethasone administered repeatedly during 3 days (subcutaneous) before the LPS challenge prevented both early events such as the macrophage priming and the TNF-alpha appearance and later events such as extracellular phospholipase A2 release and neutrophil recruitment.

Assessment of intracardiac masses by transesophageal echocardiography.

Transesophageal echocardiography and 2-dimensional transthoracic echocardiography have proved to be extremely valuable in the diagnosis of cardiac masses. In this report, we review the echocardiographic findings, clinical history, and histopathologic findings in 21 patients with intracardiac masses who underwent transthoracic echocardiography, transesophageal echocardiography, or both, at our institution. Of these patients, 14 had benign masses and 7 had malignant tumors. The potential role of transesophageal echocardiography in the diagnosis and treatment of patients with intracardiac masses is discussed. We believe that transesophageal echocardiography offers the cardiologist and cardiovascular surgeon the capability of more accurate preoperative and intraoperative assessment of cardiac masses.

A novel beta-globin mutation, beta Durham-NC [beta 114 Leu-->Pro], produces a dominant thalassemia-like phenotype.

Mutations within exon 3 of the beta-globin gene are relatively uncommon, and many of these mutations produce a dominant thalassemia-like phenotype. We describe a novel thalassemic hemoglobinopathy caused by a single nucleotide substitution (CTG-->CCG) at codon 114 resulting in a leucine to proline substitution and designate it beta Durham-NC [beta 114 Leu-->Pro]. The mutation producing this thalassemic hemoglobinopathy is located near to the beta Showa-Yakushiji mutation (beta 110 Leu-->Pro). Both of these hemoglobinopathies share similar phenotypic features with moderately severe microcytic anemia. Using computer imaging of the hemoglobin molecule, we examined several reported point mutations within exon 3 of the beta-globin gene. These point mutations cause a single amino acid substitution in the G helix, and result in a thalassemic and/or hemolytic phenotype. Computer imaging of nine separate examples suggests that amino acid substitutions affecting side chains that project into the heme pocket may destabilize the heme moiety within the beta-globin chain, resulting in a thalassemic phenotype. Hemolytic phenotypes may be the result of decreased alpha 1 beta 1 interactions. The beta Durham-NC mutation further characterizes a novel group of thalassemias/hemoglobinopathies that are clinically difficult to identify and require accessory laboratory testing.

Doppler echocardiography in 180 normally functioning St. Jude Medical aortic valve prostheses. Early and late postoperative assessments.

Limited Doppler echocardiographic data are available regarding velocities and gradients across normally functioning St. Jude Medical valves in the aortic position. To establish a standard reference for Doppler characteristics of normal aortic St. Jude Medical prostheses, we recorded continuous-wave Doppler measurements of peak and mean velocities and peak and mean gradients in 180 patients with normally functioning St. Jude aortic valves. There were 119 men and 61 women in the study; the mean age was 57 years. Minimal valvular regurgitation was present in 56 patients (31 percent). Velocities and gradients were reported in five patient groups according to valve sizes of 19 mm, 21 mm, 23 mm, 25 mm, and 27 mm; the mean gradients were 16 +/- 6, 16 +/- 6, 14 +/- 5, 12 +/- 5, and 12 +/- 6, respectively. Differences in velocities and gradients among the five valve sizes were not statistically significant (p = 0.05). Velocities and gradients were also analyzed in three patient groups according to time intervals after valve replacement. The first group (n = 64) underwent Doppler evaluation one to seven days postoperatively (mean, six days); the second group (n = 60) was evaluated after 8 to 30 days postoperatively (mean, 12 days); and the third group (n = 56) was evaluated after more than 30 days postoperatively (mean, 691 days). There were no significant differences in measurements for the three groups.