Purification and characterization of a thermostable alpha-amylase produced by the fungus Paecilomyces variotii

An alpha-amylase produced by Paecilomyces variotii was purified by DEAE-cellulose ion exchange chromatography, followed by Sephadex G-100 gel filtration and electroelution. The alpha-amylase showed a molecular mass of 75 kDa (SDS-PAGE) and pl value of 4.5. Temperature and pH optima were 60 degrees C and 4.0, respectively. The enzyme was stable for 1 h at 55 degrees C, showing a t(50) of 53 min at 60 degrees C. Starch protected the enzyme against thermal inactivation. The a-amylase was more stable in alkaline pH. It was activated mainly by calcium and cobalt, and it presented as a glycoprotein with 23% carbohydrate content. The enzyme preferentially hydrolyzed starch and, to a lower extent, amylose and amylopectin. The K(m) of alpha-amylase on Reagen (R) and Sigma (R) starches were 4.3 and 6.2 mg/mL, respectively. The products of starch hydrolysis analyzed by TLC were oligosaccharides such as maltose and maltotriose. The partial amino acid sequence of the enzyme presented similarity to alpha-amylases from Bacillus sp. These results confirmed that the studied enzyme was an a-amylase ((1 -> 4)-alpha-glucan glucanohydrolase). (C) 2010 Elsevier Ltd. All rights reserved.

Left Atrial Function After Ablation for Paroxysmal Atrial Fibrillation

Radiofirequency ablation of the pulmonary veins has been used to treat patients with paroxysmal atrial fibrillation (AF), and atrial damage after ablation is an issue of concern. To evaluate left atrial function shortly and midterm after ablation, 33 consecutive patients with paroxysmal AF were studied at baseline, 24 hours, and >= 6 months after ablation. Patients in sinus rhythm with normal ventricular function were included in the study. Echocardiographic measurements of left atrial volumes (Simpson`s rule) and transmitral and tissue Doppler myocardial (A`) velocities at the septal and lateral mitral annulus were undertaken at each time. Left atrial emptying fraction (EF; maximal - minimal left atrial volume/maximal left atrial volume) was used to express left atrial function. After 8 +/- 2 months, 30 of 33 patients returned (23 men, age 53 +/- 13 years), and all except 2 were in sinus rhythm. Shortly after ablation, left atrial minimal volumes increased (from 30 +/- 15 to 35 +/- 15 ml; p = 0.02), with maximal volumes unchanged, resulting in decreased left atrial EF (from 47 +/- 8 to 40 +/- 7 ml; p <0.05). Tissue Doppler septal A` velocities also decreased (from 8.2 +/- 1.8 to 6.9 +/- 2.0 cm/s; p <0.05). However, after midterm follow-up, both left atrial EF and septal A` velocities had slightly increased compared with shortly after ablation, although left atrial volumes remained similar to baseline. Septal A` velocity changes paralleled left atrial EF both shortly (r = 0.46, p = 0.02) and at midterm after ablation (r = 0.47, p = 0.01). In conclusion, after radiofrequency ablation, patients with paroxysmal AF experienced an initial impairment in atrial function, with improvement at longer term follow-up. (C) 2009 Elsevier Inc. All rights reserved. (Am J Cardiol 2009;103: 395-398)

Consistency of FDG-PET Accuracy and Cost-Effectiveness in Initial Staging of Patients With Hodgkin Lymphoma Across Jurisdictions

Introduction: Two hundred ten patients with newly diagnosed Hodgkin`s lymphoma (HL) were consecutively enrolled in this prospective trial to evaluate the cost-effectiveness of fluorine-18 ((18)F)-fluoro-2-deoxy-D-glucose-positron emission tomography (FDG-PET) scan in initial staging of patients with HL. Methods: All 210 patients were staged with conventional clinical staging (CCS) methods, including computed tomography (CT), bone marrow biopsy (BMB), and laboratory tests. Patients were also submitted to metabolic staging (MS) with whole-body FDG-PET scan before the beginning of treatment. A standard of reference for staging was determined with all staging procedures, histologic examination, and follow-up examinations. The accuracy of the CCS was compared with the MS. Local unit costs of procedures and tests were evaluated. Incremental cost-effectiveness ratio (ICER) was calculated for both strategies. Results: In the 210 patients with HL, the sensitivity for initial staging of FDG-PET was higher than that of CT and BMB in initial staging (97.9% vs. 87.3%; P < .001 and 94.2% vs. 71.4%, P < 0.003, respectively). The incorporation of FDG-PET in the staging procedure upstaged disease in 50 (24%) patients and downstaged disease in 17 (8%) patients. Changes in treatment would be seen in 32 (15%) patients. Cumulative cost for staging procedures was $3751/patient for CCS compared to $5081 for CCS + PET and $4588 for PET/CT. The ICER of PET/CT strategy was $16,215 per patient with modified treatment. PET/CT costs at the beginning and end of treatment would increase total costs of HL staging and first-line treatment by only 2%. Conclusion: FDG-PET is more accurate than CT and BMB in HL staging. Given observed probabilities, FDG-PET is highly cost-effective in the public health care program in Brazil.

The effect of Sao Paulo`s smoke-free legislation on carbon monoxide concentration in hospitality venues and their workers

Background Studies have shown that there is no safe level of secondhand smoke (SHS) exposure and there is a close link between SHS and the risk of coronary heart disease and stroke. Carbon monoxide (CO) is one of the most important components present in SHS. Objective To evaluate the impact of the smoking ban law in the city of Sao Paulo, Brazil, on the CO concentration in restaurants, bars, night clubs and similar venues and in their workers. Methods In the present study we measured CO concentration in 585 hospitality venues. CO concentration was measured in different environments (indoor, semi-open and open areas) from visited venues, as well as, in the exhaled air from approximately 627 workers of such venues. Measurements were performed twice, before and 12 weeks after the law implementation. In addition, the quality of the air in the city during the same period of our study was verified. Results The CO concentration pre-ban and pot-ban in hospitality venues was indoor area 4.57 (3.70) ppm vs 1.35 (1.66) ppm (p<0.0001); semi-open 3.79 (2.49) ppm vs 1.16 (1.14) ppm (p<0.0001); open area 3.31 (2.2) ppm vs 1.31 (1.39) ppm (p<0.0001); smoking employees 15.78 (9.76) ppm vs 11.50 (7.53) ppm (p<0.0001) and non-smoking employees 6.88 (5.32) ppm vs 3.50 (2.21) ppm (p<0.0001). The average CO concentration measured in the city was lower than 1 ppm during both pre-ban and post-ban periods. Conclusion Sao Paulos smoking-free legislation reduced significantly the CO concentration in hospitality venues and in their workers, whether they smoke or not.

Expression of heterochromatin protein 1 in the primary tumor of breast cancer patients in the presence or absence of occult metastatic cells in the bone marrow

Gene silencing may occur in breast cancer samples from patients presenting with occult metastatic cells in the bone marrow and one mechanism regulating gene suppression is heterochromatin formation. We have studied whether members of the heterochromatin protein 1 family Hp1(Hs alpha), Hp1(Hs beta) and Hp1(Hs gamma) which take part in chromatin packaging and gene expression regulation, were differentially expressed in tumors from patients with and without occult metastatic cells in their bone marrow. Tumor samples and bone marrow aspirates were obtained from 37 breast cancer patients. Median age was 63 years and 68% of the patients presented with clinical stage I/II disease. Presence of occult metastatic cells in bone marrow was detected through keratin-19 expression by nested RT-PCR in samples from 20 patients (54.1%). The presence of occult metastatic cells in bone marrow was not associated with node involvement, histological grade, estrogen receptor and ERBB2 immunoexpression. Relative gene expression of HP1(Hs alpha), HP1(Hs beta) and HP1(Hs gamma) was determined by real-time RT-PCR and did not vary according to the presence of occult metastatic cells in bone marrow. In addition, the combined expression of these three transcripts could not be used to classify samples according to the presence of bone marrow micrometastasis. Our work indicates that regulation of heterochromatin formation through HP1 family members may not be the sole mechanism implicated in the metastatic process to the bone marrow. (Int J Biol Markers 2008; 23: 219-24)

A standardized measurement technique may improve the reliability of measurements of endometrial thickness and volume

Objective To investigate whether standardization of the multiplanar view (SMV) when evaluating the uterus using three-dimensional ultrasonography (3D-US) improves intra-and interobserver reliability and agreement with regard to endometrial measurement. Methods Two-dimensional (2D) and 3D-US was used to measure endometrial thickness by two observers in 30 women undergoing assisted reproduction treatment. Endometrial volume was measured with Virtual Organ Computer-aided AnaLysis (VOCAL (TM)) in the longitudinal (A) and coronal (C) planes using an unmodified multiplanar view (UMV) and a standardized multiplanar view (SMV). Measurement reliability was evaluated by intraclass correlation coefficient (ICC) and agreement was examined using Bland-Altman plots with limits of agreement (LoA). The ease of outlining the endometrial-myometrial interface was compared between the A-and C-planes using subjective assessment. Results Endometrial volume measurements using the SMV and A-plane were more reliable (intra-and interobserver ICCs, 0.979 and 0.975, respectively) than were measurements of endometrial thickness using 2D-US (intra-and interobserver ICCs, 0.742 and 0.702, respectively) or 3D-US (intra-and interobserver ICCs, 0.890 and 0.784, respectively). The LoAs were narrower for SMV than for UMV. Reliability and agreement were not much different between the A- and C-planes. However the observers agreed that delineating the endometrial-myometrial interface using the A-plane was easier (first and second observer, 50.0 and 46.7%, respectively) or `comparable` (50 and 53.3%, respectively), but never more difficult than using the C-plane. Conclusions Endometrial volume measurements are more reliable than endometrial thickness measurements and are best performed using SMV and the A-plane. Copyright (C) 2011 ISUOG. Published by John Wiley & Sons, Ltd.

Gravitational field of a straight cosmic string in the framework of higher-derivative gravity

The gravitational properties of a straight cosmic string are studied in the linear approximation of higher-derivative gravity. These properties are shown to be very different from those found using linearized Einstein gravity: there exists a short range gravitational (anti-gravitational) force in the nonrelativistic limit; in addition, the derection angle of a light ray moving in a plane orthogonal to the string depends on the impact parameter.

ACE inhibition by astilbin isolated from Erythroxylum gonocladum (Mart.) OE Schulz

Erythroxylum species have several traditional uses in different countries, including the treatment of hypertension. The ethanol extract from E. gonocladum aerial parts, a species endemic to the Brazilian cerrado, elicited a concentration-dependent inhibition of angiotensin converting enzyme (ACE) (pIC(50)=4.53 +/- 0.05). Extract fractionation led to the isolation of two compounds, whose structures were assigned by spectrometric data as astilbin and beta-sitosterol, along with a mixture of palmitic, stearic and linolenic acids. This is the first report on the occurrence of these compounds on E. gonocladum. Astilbin promoted significant ACE inhibition in vitro (pIC(50)=5.86 +/- 0.33) and its activity did not differ from captopril, when both compounds were assayed at 10 mu M concentration. (C) 2009 Elsevier GmbH. All rights reserved.

Transiting exoplanets from the CoRoT space mission X. CoRoT-10b: a giant planet in a 13.24 day eccentric orbit

Context. The space telescope CoRoT searches for transiting extrasolar planets by continuously monitoring the optical flux of thousands of stars in several fields of view. Aims. We report the discovery of CoRoT-10b, a giant planet on a highly eccentric orbit (e = 0.53 +/- 0.04) revolving in 13.24 days around a faint (V = 15.22) metal-rich K1V star. Methods. We used CoRoT photometry, radial velocity observations taken with the HARPS spectrograph, and UVES spectra of the parent star to derive the orbital, stellar, and planetary parameters. Results. We derive a radius of the planet of 0.97 +/- 0.07 R(Jup) and a mass of 2.75 +/- 0.16 M(Jup). The bulk density,rho(p) = 3.70 +/- 0.83 g cm(-3), is similar to 2.8 that of Jupiter. The core of CoRoT-10b could contain up to 240 M(circle plus) of heavy elements. Moving along its eccentric orbit, the planet experiences a 10.6-fold variation in insolation. Owing to the long circularisation time, tau(circ) > 7 Gyr, a resonant perturber is not required to excite and maintain the high eccentricity of CoRoT-10b.

BthMP: a new weakly hemorrhagic metalloproteinase from Bothrops moojeni snake venom

In this work, a new weakly hemorrhagic metalloproteinase (BthMP) was purified from Bothrops moojeni snake venom. This enzyme was homogeneous by native and SDS-PAGE. It showed a polypeptide chain of 23.5 kDa, pI=7.1, and N-terminal blocked. BthMP is comprised of high proteolytic activity on casein, fibrin and bovine fibrinogen, with no coagulating, esterase or phospholipase A(2) activities; it was inhibited by EDTA, EGTA and 1,10-phenanthroline and maintained its activity on pH from 7.0 to 9.0 and temperature from 5-40 degrees C. Assays with metal ions showed that Ca(2+) is an activator, whereas Zn(2+) and Hg(2+) inhibited about 50 and 80% of its activity, respectively. The edema evidenced the important role of the toxin in the inflammatory activity of the venom. BthMP also caused unclotting, and provoked histological alterations in the gastrocnemius muscle of mice inducing hemorrhage, necrosis and leukocytic infiltrate. The molecular mass and the inhibition assays suggest that the metal loproteinase BthMP belongs to class P-I of SVMPs. (c) 2008 Elsevier Ltd. All rights reserved.

Bacterial colonization in hemodialysis temporary dual lumen catheters: A prospective study

Aims. The use of hemodialysis temporary dual-lumen catheters is often complicated by infections, which may be a significant cause of death among patients with end stage renal disease (ESRD). The aim of this study was to assess the incidence of bacteremia and bacterial colonization related to non-tunneled, non-cuffed, dual-lumen temporary catheters in patients with ESRD submitted to hemodialysis. Methods. This study included 29 patients with ESRD. After catheter implantation, patients were monitored throughout the period of catheter permanence by means of blood samples collected weekly from a peripheral vein. Bacteria were isolated and identified according to CLSI recommendations. When catheters were removed for any reason, their tips were evaluated microbiologically. Results. A total of 194 blood samples from the 29 patients implanted with 55 catheters were analyzed. Of these, 15.5% (30 samples) demonstrated bacterial growth, principally Staphylococcus epidermidis (64.5%). Twenty patients (68.9%) presented at least one positive blood culture during follow-up. The median time for catheter colonization was 18.5 days (95% CI: 16.8-30.3). Of the 55 catheters implanted, 28 (50.9%) showed bacterial colonization, corresponding to 23.4 episodes/1000 catheter/days and 9.2 episodes of bacteremia /1000 catheter/days. Fifteen of 28 catheter tips analyzed showed bacterial growth (53.5%). In 14 of these (93.3%), there was agreement between the isolates from the catheter tip and blood cultures. Of 24 episodes of positive blood cultures from 20 different patients in 17 episodes (70.8%), the patients showed no clinical signs or symptoms of bacteremia. Conclusions. The high incidence of catheter colonization, the correlation between blood and catheter tip cultures, and the occurrence of frequent cases of asymptomatic bacteremia justify the proposal of routine peripheral blood collections to monitor patients undergoing hemodialysis with temporary dual-lumen catheters.

Influence of anesthetics on pharyngeal flap surgery: a 23-year study

This study assessed the influence of anesthetics on early complications after pharyngeal flap surgery. A 23-year retrospective chart review was carried out of all patients at the authors` institution who underwent superiorly based pharyngeal flap surgery. Variables analyzed were gender, age at the time of surgery, cleft type, anesthesia procedure used and complications in the early postoperative period. 2299 patients (50% male; 50% female) who underwent pharyngeal flap surgery between 1980 and 2003 were reviewed. The highest number of surgeries was performed in patients aged 11-20 years. There were 1042 patients with at least one type of complication. Of these, 39 required reoperation to control complications such as bleeding and airway obstruction. There were no records of death. Vomiting and pain were the most frequent postoperative complications (16% and 14% of patients, respectively). Lower complication rates were observed when anesthesia protocols included sevoflurane, propofol and opioids.