Preclinical assessment of an anti-HTLV-1 heterologous DNA/MVA vaccine protocol expressing a multiepitope HBZ protein.

BACKGROUND: Human T-lymphotropic virus 1 (HTLV-1) is associated with the development of several pathologies and chronic infection in humans. The inefficiency of the available treatments and the challenge in developing a protective vaccine highlight the need to produce effective immunotherapeutic tools. The HTLV-1 basic leucine zipper (bZIP) factor (HBZ) plays an important role in the HTLV-1 persistence, conferring a survival advantage to infected cells by reducing the HTLV-1 proteins expression, allowing infected cells to evade immune surveillance, and enhancing cell proliferation leading to increased proviral load. METHODS: We have generated a recombinant Modified Virus Vaccinia Ankara (MVA-HBZ) and a plasmid DNA (pcDNA3.1(+)-HBZ) expressing a multiepitope protein based on peptides of HBZ to study the immunogenic potential of this viral-derived protein in BALB/c mice model. Mice were immunized in a prime-boost heterologous protocol and their splenocytes (T CD4+ and T CD8+) were immunophenotyped by flow cytometry and the humoral response was evaluated by ELISA using HBZ protein produced in prokaryotic vector as antigen. RESULTS: T CD4+ and T CD8+ lymphocytes cells stimulated by HBZ-peptides (HBZ42-50 and HBZ157-176) showed polyfunctional double positive responses for TNF-α/IFN-γ, and TNF-α/IL-2. Moreover, T CD8+ cells presented a tendency in the activation of effector memory cells producing granzyme B (CD44+High/CD62L-Low), and the activation of Cytotoxic T Lymphocytes (CTLs) and cytotoxic responses in immunized mice were inferred through the production of granzyme B by effector memory T cells and the expression of CD107a by CD8+ T cells. The overall data is consistent with a directive and effector recall response, which may be able to operate actively in the elimination of HTLV-1-infected cells and, consequently, in the reduction of the proviral load. Sera from immunized mice, differently from those of control animals, showed IgG-anti-HBZ production by ELISA. CONCLUSIONS: Our results highlight the potential of the HBZ multiepitope protein expressed from plasmid DNA and a poxviral vector as candidates for therapeutic vaccine.

Transmission of intra-cellular genetic information: a system proposal.

One of the great challenges of the scientific community on theories of genetic information, genetic communication and genetic coding is to determine a mathematical structure related to DNA sequences. In this paper we propose a model of an intra-cellular transmission system of genetic information similar to a model of a power and bandwidth efficient digital communication system in order to identify a mathematical structure in DNA sequences where such sequences are biologically relevant. The model of a transmission system of genetic information is concerned with the identification, reproduction and mathematical classification of the nucleotide sequence of single stranded DNA by the genetic encoder. Hence, a genetic encoder is devised where labelings and cyclic codes are established. The establishment of the algebraic structure of the corresponding codes alphabets, mappings, labelings, primitive polynomials (p(x)) and code generator polynomials (g(x)) are quite important in characterizing error-correcting codes subclasses of G-linear codes. These latter codes are useful for the identification, reproduction and mathematical classification of DNA sequences. The characterization of this model may contribute to the development of a methodology that can be applied in mutational analysis and polymorphisms, production of new drugs and genetic improvement, among other things, resulting in the reduction of time and laboratory costs.

Comparative Analysis of Gait Speed Estimation Using Wideband and Narrowband Radars, Thermal Camera, and Motion Tracking Suit Technologies.

Research has shown that cognitive and physical functioning of older adults can be reflected in indicators such as walking speed. While changes in cognition, mobility, or health cause changes in gait speed, often gradual variations in walking speed go undetected until severe problems arise. Discrete clinical assessments during clinical consultations often fail to detect changes in day-to-day walking speeds and do not reflect walking speeds in everyday environments, where most of the mobility issues happen. In this paper, we compare four walking speed measurement technologies to a GAITRite mat (gold standard): (1) an ultra wideband radar (covering the band from 3.3 GHz to 10 GHz), (2) a narrow band 24-GHz radar (with a bandwidth of 250 MHz), (3) a perception Neuron Motion Tracking suit, and (4) a thermal camera. Data were collected in parallel using all sensors at the same time for 10 healthy adults for normal and slow walking paces. A comparison of the sensors indicates better performance at lower gait speeds, with offsets (when compared to GAITRite) between 0.1 and 20% for the ultra wideband radar, 1.9 and 17% for the narrowband radar, 0.1 and 38% for the thermal camera, and 1.7 and 38% for the suit. This paper supports the potential of unobtrusive radar-based sensors and thermal camera technologies for ambient autonomous gait speed monitoring for contextual, privacy-preserving monitoring of participants in the community.

Sodium chloride and water transport in the medullary thick ascending limb of Henle. Evidence for active chloride transport.

Transport of NaCl and water was examined in the rabbit medullary thick ascending limb of Henle (ALH) by perfusing isolated segments of these nephrons in vitro. Osmotic water permeability was evaluated by perfusing tubules against imposed osmotic gradients. In these experiments the net transport of fluid remained at zero when segments of thick ALH were perfused with isotonic ultrafiltrate in a bath of rabbit serum in which the serum osmolality was increased by the addition of either 239+/-8 mosmol/liter of raffinose or 232+/-17 mosmol of NaCl indicating that the thick ascending limb of Henle is impermeant to osmotic flow of water. When these tubules were perfused at slow rates with isosmolal ultrafiltrate of same rabbit serum as used for the bath, the effluent osmolality was consistently lowered to concentrations less than the perfusate and the bath. That this decrease in collected fluid osmolality represented salt transport was demonstrated in a separate set of experiments in which it was shown that the sodium and chloride concentrations decreased to 0.79+/-0.02 and 0.77+/-0.02 respectively when compared with the perfusion fluid concentrations. In each instance the simultaneously determined transtubular potential difference (PD) revealed the lumen to be positive with the magnitude dependent on the perfusion rate. At flow rates above 2 nl.min(-1), the mean transtubular PD was stable and equal to 6.70+/-0.34 mv. At stop-flow conditions this PD became more positive. Ouabain and cooling reversibly decreased the magnitude of this PD. The transtubular PD remained positive, 3.3+/-0.2 mV, when complete substitution of Na by choline was carried out in both the perfusion fluid and the bathing media. These results are interpreted to indicate that the active transport process is primarily an electrogenic chloride mechanism. The isotopic permeability coefficient for Na was 6.27+/-0.38 x 10(-5) cm.s(-1) indicating that the thick ALH is approximately as permeable to Na as the proximal convoluted tubule. The chloride permeability coefficient for the thick ALH was 1.06+/-0.12 x 10(-5) cm.s(-1) which is significantly less than the chloride permeability of the proximal tubule. These data demonstrate that the medullary thick ascending limb of Henle is water impermeable while having the capacity for active outward solute transport as a consequence of an electrogenic chloride pump. The combination of these characteristics allows this segment to generate a dilute tubular fluid and participate as the principal energy source for the overall operation of the countercurrent multiplication system.

Calcium and phosphate transport in isolated segments of rabbit Henle's loop.

Calcium and phosphate transport was examined in rabbit thin descending, thin ascending, and thick ascending limbs of Henle by in vitro perfusion of isolated tubular segments. Permeability coefficients for these segments with 45Ca and 32PO4 were determined for both lumen-to-bath and bath-to-lumen directions. Both the thin descending and thin ascending limbs were found to be relatively impermeable to both 45Ca and 32PO4. In neither segment were we able to show evidence for net transport of calcium or phosphate. In contrast, the thick ascending limb of Henle showed a decrease in calcium lumen-to-bath concentration from 0.97 +/- 0.02 to 0.88 +/- 0.02 when perfused at 4.8 nl min-1. 45Ca lumen-to-bath and bath-to-lumen fluxes were 19.96 +/- 1.05 and 9.89 +/- 0.02 peq-min-1-cm-1, respectively, and the potential difference was +3.8 +/- 0.3 mV (lumen positive). The observed calcium flux ratio was significantly higher than that predicted by Ussing's equation. When ouabain was added to the bath the potential difference fell to +1.1 +/- 0.3 mV, whereas the calcium efflux was only slightly diminished (29.5 +/- 5.3-23.7 +/- 5.1 peq-cm-1-min-1). Ouabain had no effect on the influx of Ca across the thick ascending limb of Henle. There was no net transport of phosphate across the thick ascending limb. Phosphate permeability was exceedingly low bidirectionally across the thick ascending limb. Our findings indicate: (a) all segments of Henle's loop are relatively impermeable to calcium and phosphate; (b) net transport of phosphate seems to be absent in Henle's loop; (c) net calcium reabsorption, which cannot be explained by passive mechanisms, occurs in the thick ascending limb.

Defective E-cadherin/catenin complexes in human cancer.

Cancer is caused by a series of genomic changes leading directly or indirectly to disturbance of growth, differentiation and tissue integrity. Genomic, transcriptional or posttranscriptional alterations of E-cadherin/catenin complexes that are implicated in various steps of cancer development comprise mutational inactivation, transcriptional downregulation of E-cadherin sometimes accompanied by upregulation of N-cadherin, proteolysis of E-cadherin and posttranslational stabilisation of beta-catenin and plakoglobin. The E-cadherin/catenin complex serves not only cell-cell adhesion but also transduces signals to the nucleus and to the cytoskeleton, either directly or through its connections with multiple other complexes. We review here the expression of E-cadherin/catenin in human cancers, emphasising methods of observation and prognostic interpretation of results. This is illustrated in thyroid lesions from the benign follicular adenoma to the extremely malignant anaplastic carcinoma. The eye is an organ largely neglected by students of cadherins and catenins. The implication of a variety of members of these molecular families in the embryonic development of the eye strongly suggests that disturbances of cadherin/catenin complexes are crucial also in the development of ocular tumours.

Calcium transport in isolated perfused pars recta of proximal tubule.

1. Ca2+ transport was examined in segments of rabbit pars recta isolated from the renal cortex by the technique of in vitro perfusion. 2. The bidirectional Ca2+ flux was determined by adding 45Ca to the bath and to the perfusate sequentially. Five tubules were perfused with Ringer-HCO-3 solution containing 0.5% albumin and bathed in the same solution, which maintained the ionized Ca2+ at the same concentration in the perfusate and bath. The outflux of Ca2+ was 169 +/- 18 pEq cm-1 min-1, while the influx was 53 +/- 13 pEq cm-1 min-1. The difference between these two numbers, the net Ca2+ absorption, is equal to 116 +/- 19 pEq cm-1 min-1. The flux ratio, 3.79 +/- 0.87, was significantly higher than 0.91 +/- 0.01 predicted by the Ussing equation from the transtubular potential difference (PD = -1.3 +/- 0.2 mV). 3. The effect of ouabain and transtubular PD on Ca2+ outflux was studied with rabbit serum ultrafiltrate as perfusate and rabbit serum as the bath. When 10(-5) M ouabain was added to the bath the transtubular PD fell from -2.4 +/- 0.3 mV to -0.65 +/- 0.06mV and fluid absorption decreased from 0.83 +/- 0.15 to 0.42 +/- 0.2 nl min-1 mm-1, whereas Ca2+ outflux remained unchanged (before = 160 +/- 13 pEq cm-1 min-1, after = 167 +/- 7 pq cm-1 min-1). 4. When tubules were perfused with and Na+ -free solution (NaCl replaced by choline chloride), Ca2+ outflux, 170 +/- 170 11 pEq cm-1 min-1, was the same as that obtained with the normal perfusion solution. 5. These results indicate that: a) the pars recta has the capacity for net absorption of significant amounts of Ca2+ that is apparently independent of Na+ efflux; b)Ca2+ transport across the pars recta occurs by a mechanism other than simple diffusion and c), unlike Na+ transport, Ca2+ outflux is not sensitive to ouabain.

[Coronary artery dissection without acute myocardial infarction]. / Dissecção de Artéria coronária sem infarto agudo do miocárdio.

A 57 year-old-man with acute aortic dissection (DeBakey type I) who developed right coronary artery dissection without acute myocardial infarction. He was successful surgically treated and became asymptomatic.

[Myocardial revascularization in left coronary trunk lesion in patients over 65 years old]. / Cirurgia de revascularização miocárdica na lesão do tronco da coronária esquerda em pacientes acima de 65 anos de idade.

PURPOSE: To study the short and long-term follow-up of patients with left main coronary artery disease (LMCAD) and age over 65 years, by comparing the results with patients under 65 years-old. METHODS: Twenty-two patients with LMCAD and mean age of 69 +/- 3.5 years (group I) were underwent isolated coronary artery bypass grafting (CABG) and compared to 31 patients with LMCAD, mean age of 54 +/- 7 years (group II), who also underwent isolated CABG. The life-table Kaplan-Meyer method was used to estimate the post-operative survival. The chi-square and Student "t" test were used when necessary. RESULTS: Despite higher operative mortality in group I (9.1% x 3.2%), the difference was statistically not significant. The operative morbidity was similar in both groups. Actuarial survival at 4 years was 85% in group I and 95% in group II. Actuarial survival free of cardiac events was 69% in group II and 75% in group II. CONCLUSION: The CABG is well tolerated and had low morbidity and acceptable mortality in old patients with LMCAD. The long-term survival in these patients was very similar to the younger patients.

Aortic stenosis. Gender influence on left ventricular geometry and function in patients under 70 years of age.

OBJECTIVE: To verify if adaptive left ventricle (LV) characteristics are also present in individuals under 70 years of age with severe aortic stenosis (AS). METHODS: The study comprised 40 consecutive patients under 70 years of age with AS and no associated coronary artery disease, referred for valve surgery. Out of the 40 patients, 22 were men and 18 women, and the mean age was 49.8 +/- 14.3 years. Cardiac symptoms, presence of systemic hypertension (SH), functional class according to the New York Heart Association (NYHA), and valve lesion etiology were considered. LV cavity dimensions, ejection fraction (EF), fractional shortening (FS), mass (MS), and relative diastolic thickness (RDT) were examined by Doppler echocardiography. RESULTS: Fourteen (63.6%) men and 11 (61.6%) women were classified as NYHA class III/IV (p = 0.70). There was no difference in the frequency of angina, syncope or dyspnea between genders. The incidence of SH was greater in women than in men (10 versus 2, p = 0.0044). Women had a smaller LV end-diastolic diameter index (32.1 +/- 6.5 x 36.5 +/- 5.3 mm/m2, p = 0.027), LV end-systolic diameter index (19.9 +/- 5.9 x 26.5 +/- 6.4 mm/m2, p = 0.0022) and LV mass index (MS) (211.4 +/- 71.1 x 270.9 +/- 74.9 g/m2, p = 0.017) when compared with men. EF (66.2 +/- 13.4 x 52.0 +/- 14.6%, p = 0.0032), FS (37.6 +/- 10.7 x 27.9 +/- 9.6%, p = 0.0046) and RDT (0.58 +/- 0.22 x 0.44 +/- 0.09, p = 0.0095) were significantly greater in women than in men. CONCLUSION: It is the patient gender rather than age that influences left ventricular adaptive response to AS.

Myxoma of the mitral valve.

Only rarely do myxomas originate from the mitral valve. This is the report of a 49-year-old woman presenting with congestive heart failure. The diagnosis of an intracardiac tumor involving the anterior cuspid of the mitral valve was made by transesophageal echocardiography. The patient underwent surgery for tumor resection and plasty of the valve was made with reconstruction and preservation of the valve. The diagnosis of myxoma was confirmed by histology. This is the 23rd case of myxoma of the mitral valve reported in the literature.

[Silent myocardial ischemia. Experience with ambulatory electrocardiographic monitoring]. / Isquemia miocárdica silenciosa. Experiência com a moniorização eletrocardiográfica ambulatoriai.

A 24-hour recording of ambulatory EKG (Holter) was obtained in 159 patients with coronary disease: previous myocardial infarction (n = 76), myocardial revascularization (n = 66), severe obstruction during cine-coronarography (n = 13) or during coronary angioplasty (n = 4). Prophylactic anti-anginal drugs were maintained during the Holter and, despite the medication, transitory episodes of myocardial ischemia (MI) were recorded in 51 patients (32%), either with angina (AMI) or silent (SMI), isolated SMI was recorded in 44 patients (86%), 6 subjects had both SMI (16 episodes) and AMI (12 episodes) and one patient had only one episode of AMI. There was a total of 119 episodes of MI, 106 of SMI (89.1%) and 13 AMI (10.9%). The total duration of the episodes of SMI per patient varied from 1 min to 235 min and the IMS maximal duration was 221 min. In the six cases with both SMI and AMI, the total duration of SMI was 461 min and AMI was 306 min. The circadian distribution of the episodes of SMI was: from 12:00 to 18:00-31.4%, 18:00 to 24:00-27.6%, 0:00 to 6:00-7.7% and 6:00 to 12:00-33.3%. The SMI activity was recorded in 89 episodes-23.6% during sleep, 22.4% at rest, 46% during physical activity and 8% during other activities. The mean ST segment depression during SMI (n = 106) was of -2.25 mm and during AMI (n = 13) was of -3.25 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

Caffeine triggers behavioral and neurochemical alterations in adolescent rats.

Caffeine is the psychostimulant most consumed worldwide but concerns arise about the growing intake of caffeine-containing drinks by adolescents since the effects of caffeine on cognitive functions and neurochemical aspects of late brain maturation during adolescence are poorly known. We now studied the behavioral impact in adolescent male rats of regular caffeine intake at low (0.1mg/mL), moderate (0.3mg/mL) and moderate/high (1.0mg/mL) doses only during their active period (from 7:00 P.M. to 7:00 A.M.). All tested doses of caffeine were devoid of effects on locomotor activity, but triggered anxiogenic effects. Caffeine (0.3 and 1mg/mL) improved the performance in the object recognition task, but the higher dose of caffeine (1.0mg/mL) decreased the habituation to an open-field arena, suggesting impaired non-associative memory. All tested doses of caffeine decreased the density of glial fibrillary acidic protein and synaptosomal-associated protein-25, but failed to modify neuron-specific nuclear protein immunoreactivity in the hippocampus and cerebral cortex. Caffeine (0.3-1mg/mL) increased the density of brain-derived neurotrophic factor (BDNF) and proBDNF density as well as adenosine A1 receptor density in the hippocampus, whereas the higher dose of caffeine (1mg/mL) increased the density of proBDNF and BDNF and decreased A1 receptor density in the cerebral cortex. These findings document an impact of caffeine consumption in adolescent rats with a dual impact on anxiety and recognition memory, associated with changes in BDNF levels and decreases of astrocytic and nerve terminal markers without overt neuronal damage in hippocampal and cortical regions.

Anxiolytic-like effects of a new 1-N substituted analog of melatonin in pinealectomized rats.

In spite of the wide variety of drugs available for treating anxiety, this disorder continues to represent a worldwide health problem that is classified within the first 10 causes of disability. Therefore, the search continues for new antianxiety agents, particularly those not related to benzodiazepines. Even though melatonin has been prescribed as an anxiolytic drug, its use is currently limited due to its short half-life and photo-sensitivity, among other disadvantages. The present study explores the antianxiety properties of a new 1-N substituted melatonin analog, M3C, in pinealectomized rats submitted to two behavioral tests (the cumulative burying behavior paradigm and the elevated plus-maze). Results from both tests show that M3C is effective as an anxiolytic-like agent, at doses lower than any other melatonin analog previously reported. The blocking of these actions by luzindole together with the available data suggests that the anxiolytic properties of M3C are mediated by MT1 and MT2 receptors.

Isolation and pathogenicity of Toxoplasma gondii in naturally infected (rustic farm) pigs in southern Brazil.

This study reported a serological test for Toxoplasma gondii infection in 100 pigs from 58 rural farms in the state of Rio Grande do Sul, Brazil. Thirty-six pigs were seropositive (IFAT≥1:64). Bioassays were performed for all 36 seropositive pigs, and 17 isolates were obtained (47.2%). Seven of these isolates (41.2%) were highly pathogenic to mice, as clinical signs of acute infection were observed, and tachyzoites were found in the peritoneal exudates, livers, and lungs. The remaining 10 isolates were able to establish a chronic infection in mice, therefore, they were not highly virulent. The results of this study indicate that pork is a potential source of T. gondii transmission to humans.