Native fungi from Amazon with potential for control of Aedes aegypti L. (Diptera: Culicidae).

Aedes aegypti L. (Diptera: Culicidae) is the main transmitter of pathogens that cause human diseases, including dengue, chikungunya, zika and yellow fever. Faced with this problem, this study aims to select fungi with entomopathogenic potential against Ae. aegypti and develop formulations that optimize the control action of entomopathogenic fungi in the semi-field condition. 23 fungal strains native from Amazon were inoculated in Potato-Dextrose-Agar (PDA) culture medium for 14 days and then transferred by scraping to tubes containing 0.9% NaCl solution. To obtain the larvae, eggs were collected using traps in peridomestic environments for 7 days. 20 larvae of Ae. aegypti in 125 mL erlenmeyers containing 20 mL of conidial suspension at a concentration of 1x106 conidia/mL for initial selection and 1×104, 1×105, 1×106 and 1×107 conidia/mL for determination of LC50. Mortality was checked every 24 h for 5 days. The three fungi with the best virulence rates were identified using molecular techniques. The compatibility between fungi at a concentration of 1×106 conidia/mL and oily adjuvants, mineral oil and vegetable oil (andiroba, chestnut and copaiba) at concentrations of 0.1, 0.5 and 1% was evaluated. The germination capacity of 100 conidia per treatment was evaluated after incubation at 28 ºC for 24 h. To evaluate the entomopathogenic potential of the fungal formulations, conidial suspensions (1×106 conidia/mL) were added with 0.1% mineral and vegetable oil. The treatments were submitted to laboratory and semi-field conditions and mortality was verified every 24 h for 5 days. Beauveria sp. (4,458) (LC50 = 8.66× 103), Metarhizium anisopliae (4,420) (LC50 = 5.48×104) and M. anisopliae (4,910) (LC50 = 1.13×105) were significantly more effective in the larval control of Ae. aegypti, in relation to the other fungal morphospecies evaluated. Mineral oil was better compatible in all treatments evaluated. Beauveria sp. (4,458) was considerably less virulent under semi-field conditions. M. anisopliae (4,910) formulated with mineral oil increased larval mortality to 100% on the 4th day in the laboratory and on the 5th day in the semi-field. Fungal formulations developed from native Amazonian isolates represent a promising tool for the development of strategies to control Ae. aegypti.

Population Development of Zabrotes subfasciatus (Coleoptera: Chrysomelidae) in Landrace Bean Varieties Occurring in Southwestern Amazonia.

The common bean, Phaseolus vulgaris (L.), is one of the most important sources of protein worldwide, and Latin America is one of the recognized centers of diversity of this species. However, storage of this product after harvest is not feasible because of bruchid attacks. This study determined the accumulated normalized rate of emergence and the daily emergence rate of Zabrotes subfasciatus (Boheman) (Coleoptera: Chrysomelidae:Bruchinae) in five landrace varieties of common bean (BRL 01, SNA 01, RDR 01, RBC 01, and RBC 13) that occurin southwestern Amazonia. These varieties were selected for this study because they are well-distributed throughout the Amazonian communities. Beans of each variety were infested with 50 unsexed adults, and the insects were removed 13 d after beginning the bioassays. The adult progeny obtained from the feeding substrate were counted and removed every other day after the first emergence, until the end of the emergence period. Differences were observed in the calculated rates of development; however, the time required for development and emergence of the insects was independent. Of the five varieties of bean investigated, we observed that the RDR 01, BRL 01, and SNA 01 cultivars are resistant to Z. subfasciatus; the results indicate that the use of these three varieties can reduce problems associated with bruchid attacks and enable storage of the product after harvesting.

Immunogenic activity of the fish tapeworm Pterobothrium heteracanthum (Trypanorhyncha: Pterobothriidae) in BALB/c mice.

The aim of this study was to verify the immunogenicity of Pterobothrium heteracanthum (Cestoda: Trypanorhyncha) crude protein extract (PH-CPE) in BALB/c mice. The parasites were obtained from Micropogonias furnieri (Osteichthyes: Sciaenidae). Groups of six mice were each immunized with 10, 50 or 100 µg of PH-CPE, on days 0 and 35. Both specific IgG and IgE responses were developed after immunization. The immunoblot assay revealed that specific IgG recognizes PH-CPE proteins with two molecular weight ranges, 60-75 and 30-40 kDa, and that IgE recognizes larger proteins over 120 kDa. This appears to be the first report on the immunogenicity of metacestodes within the Pterobothriidae and that PH-CPE is a potential inducer of a specific IgE response.

Nanoparticles and surfaces presenting antifungal, antibacterial and antiviral properties.

Here, we present new antimicrobial nanoparticles based on silica nanoparticles (SNPs) coated with a quaternary ammonium cationic surfactant, didodecyldimethylammonium bromide (DDAB). Depending on the initial concentration of DDAB, SNPs immobilize between 45 and 275 µg of DDAB per milligram of nanoparticle. For high concentrations of DDAB adsorbed to SNP, a bilayer is formed as confirmed by zeta potential measurements, thermogravimetry, and diffuse reflectance infrared Fourier transform (DRIFT) analyses. Interestingly, these nanoparticles have lower minimal inhibitory concentrations (MIC) against bacteria and fungi than soluble surfactant. The electrostatic interaction of the DDAB with the SNP is strong, since no measurable loss of antimicrobial activity was observed after suspension in aqueous solution for 60 days. We further show that the antimicrobial activity of the nanoparticle does not require the leaching of the surfactant from the surface of the NPs. The SNPs may be immobilized onto surfaces with different chemistry while maintaining their antimicrobial activity, in this case extended to a virucidal activity. The versatility, relative facility in preparation, low cost, and large antimicrobial activity of our platform makes it attractive as a coating for large surfaces.

Conjoined twins pregnancies: experience with 36 cases from a single center.

OBJECTIVE: To review a single center's experience in the management of twin pregnancies with conjoined fetuses. METHODS: Retrospective study describing prenatal findings, delivery details, surgical treatment and perinatal outcome. RESULTS: The study included 36 twin pregnancies with conjoined twins seen over a period of 12 years in a single tertiary hospital: 69.4% were thoracopagus, 13.9% parapagus, 8.3% omphaloischiopagus 5.6% omphalopagus and 2.8% cephalopagus. Cardiac defects were present in 91.6% of twin pairs and associated malformations were present in 61.8% of the cases: limb abnormalities in 36.1%, abdominal wall defects in 25.0%, cleft lip and/or palate in 13.9% and congenital diaphragmatic hernia in 5.5%. Surgical separation was considered not feasible and prognosis lethal in 30 (83.3%) cases. Termination of pregnancy was performed in 12 pregnancies of poor prognosis. Cesarean section was performed in all remaining cases. Five sets of twins underwent surgical separation and six children survived. Overall survival in our series was 8.3% and, among the livebirths, 13.6%. CONCLUSION: Conjoined twin pregnancies should be referred to tertiary centers for detailed fetal anomaly and echocardiographic assessment to evaluate prognosis and determine the possibility of postnatal surgical separation.

Fusion proteins containing family 1 and family 2 PspA fragments elicit protection against Streptococcus pneumoniae that correlates with antibody-mediated enhancement of complement deposition.

PspA is an important pneumococcal vaccine candidate that is capable of inducing protection in different animal models. Because of its structural diversity, a PspA-based vaccine should contain at least one fragment from each of the two major families (1 and 2) in order to elicit broader protection. In the present work, we have tested the potential of PspA hybrids containing fused portions of family 1 and 2 (PspA1ABC-4B and PspA1ABC-3AB) PspA fragments to induce protection against pneumococci bearing distinct PspA fragments. Sera from mice immunized with these hybrid PspA fragments were able to increase C3 deposition on pneumococci bearing PspA fragments from both families, in contrast with sera made against the PspA family 1 (PspA1ABC) and PspA family 2 (PspA3ABC) fragments, which were effective only within the same family. Although PspA hybrids were able to extend protection against pneumococcal infection with strains bearing diverse PspA fragments, the immunity elicited by family 2 was clade dependent, suggesting that PspA fragments from family 2 clades 3 and 4 should both be included in a comprehensive PspA vaccine. These results indicate that PspA fusion proteins constitute an efficient immunization strategy for future PspA-based antipneumococcal vaccines since they are able to extend protection provided by a protein derived from a single transcript.

Three-dimensional ultrasound fetal urinary bladder volume measurement: reliability of rotational (VOCAL) technique using different steps of rotation.

OBJECTIVE: The aim of the study was to investigate the inter- and intraobserver reliability in measuring the fetal bladder volume by 3d ultrasound imaging by VOCAL. In addition a comparison of 15 degrees and 30 degrees rotation steps calculation had been performed. METHODS: Measurements (n = 240) of FB volume were performed in 30 low-risk patients with gestation times ranging from 22 to 39 weeks by two independent observers blind to each other. All fetuses were examined and no ultrasound detectable malformation was found. The volume for each FB was measured twice by the observers who used the Virtual Organ Computed-aided Analysis (VOCAL) technique. Distinct sets of 12 and 6 planes were obtained after sequential rotations of 15 degrees and 30 degrees, respectively. The internal contour of fetal bladder was determined manually and carefully as to exclude adjacent structures from volume calculation. Wilcoxon signed-rank test was used for the comparison of paired samples in the cases of replication within and between observers. Spearman's rank correlation was used to study the relationship among angles. Bland and Altman's graphical approach was used to investigate the agreement between observers. RESULTS: Both techniques were shown to be highly reliable. No significant difference was found between the measurements of FB volume with the VOCAL technique by varying either the steps of rotation or the observers. CONCLUSION: Excellent correlations were found for both observers in the use of rotation angles of 15 degrees and 30 degrees . Since a significantly faster evaluation was obtained by using a 30 degrees rotation step it must be preferred to assess the FB volume. Our results show that the VOCAL technique, with a plane rotation of 30 degrees , can be used in a simple way to estimate fetal urine production.

Antimycobacterial activity of some Brazilian indigenous medicinal drinks

Tuberculosis (TB) is a very serious problem worldwide and the increasing number of multiple drugs resistant TB cases makes the search for new anti-TB drugs an urgent need. Indigenous knowledge about the use of native plants to treat illnesses has contributed to the discovery of new medicines. In this study, the antimycobacterial activity ofseven medicinal drinks was assessed: Ananas sativus (hydroalcoholic fruit extract), Aristolochia triangularis(aqueous and hydroalcoholic leaf, root and stem extracts), Bromelia antiacantha (hydroalcoholic fruit extract), Stryphnodendron adstringens (hydroalcoholic bark extract), Tabebuia ovellanedae (hydroalcoholic bark extract), Vernonia polyanthes (hydroalcoholic root extract), all used by the Vanuíre indigenous community in the treatment of respiratory diseases. The activity was evaluated by using a time-to-kill assay, in which Mycobacterium tuberculosis H37Rv was cultured on Lowenstein-Jensen medium, after thirty minutes, one, three, six, twelve and twenty-four hours contact of the bacteria with each drink. Within half to one hour contact, the hydroalcoholic drinks of A. triangularis, S. adstringens, T. ovellanedae and V. polyanthes reduced the bacterial growth by 2 orders of magnitude in CFU/mL, and all bacterial growth was absent after three hours contact. In contrast, no mycobactericidal effect was detected in the aqueous extract of A. triangularis or in the hydroalcoholic beverages of A. sativus and B. antiacantha, even aftertwenty-four hours contact.

Identification of oligomers in polyethyleneterephthalate bottles for mineral water and fruit juice. Development and validation of a high-performance liquid chromatographic method for the determination of first series cyclic trimer.

Cyclic oligomers were identified in PET bottles used for mineral water and fruit juice using MS and (1)H and (13)C NMR: a first series cyclic trimer, a first series cyclic tetramer, a first series cyclic dimmer and a second series cyclic trimer. An analytical method to determine first series cyclic trimer in these bottles was developed and validated, using HPLC. The first series cyclic trimer levels were 316-462 mg/100g of PET bottle.

Ductus venosus blood flow assessment at 11 to 14 weeks of gestation and fetal outcome.

OBJECTIVE: To evaluate the association between abnormal ductus venosus (DV) at 11-14 weeks' gestation and chromosomal abnormalities, structural defects and fetal outcome. METHODS: DV flow-velocity waveform (DV-FVW) and nuchal translucency thickness (NT) were prospectively evaluated in 1217 singleton pregnancies. RESULTS: The DV-FVW was abnormal in 84 fetuses, NT was above the 95th centile in 160 fetuses and both markers were observed in 41 fetuses. Chromosomal defects were diagnosed in 22 fetuses. The sensitivity, specificity and positive and negative predictive values for an abnormal karyotype were 86.4%, 86.9%, 11.9% and 99.7%, respectively, for an increased NT. These values were 68.2%, 96.9%, 31.3% and 99.3%, respectively, for DV-FVW abnormalities and 68.2%, 97.6%, 36.6% and 99.3%, respectively, when both markers were found simultaneously. Regarding structural defects, these values were 43.8%, 92.9%, 8.3% and 99.1% for an abnormal NT, 25.0%, 92.6%, 4.8% and 98.8% for DV-FVW abnormalities and 25.0%, 97.9%, 15.4% and 98.9% for both together. Considering those cases of unexplained fetal demise, the values were 44.4%, 85.9%, 5.0% and 98.9% for NT abnormalities, 22.2%, 92.6%, 4.8% and 98.6% for an abnormal DV-FVW and 22.2%, 98%, 15.4% and 98.7% for both. In cases with increased NT, the percentage of live births with normal karyotype and no major fetal structural defects decreased from 93.8% in normal DV-FVW fetuses to 77.3% in abnormal ones. CONCLUSION: DV assessment at 11-14 weeks' gestation is useful in screening for fetal chromosomal abnormalities and may help to reduce the false-positive rate when combined with NT measurement. Abnormal DV-FVW is also associated with an increase in adverse perinatal outcome in fetuses with enlarged NT. However, the value of DV-FVW assessment in cases with normal NT is unclear.

Structural and functional cardiac abnormalities identified prior to 16 weeks' gestation in fetuses with increased nuchal translucency.

OBJECTIVE: To determine the accuracy and practicality of fetal echocardiography in the identification of structural and functional cardiac abnormalities prior to 16 weeks' gestation in fetuses with increased nuchal translucency thickness (NT). METHODS: Between January 1996 and June 2002 early fetal echocardiography using the transvaginal route was performed at 12-16 weeks' gestation on 275 fetuses with increased NT. The abnormal cardiac findings were classified as either structural (congenital heart defects) or functional, defined as transient phenomena which might later disappear, such as isolated tricuspid regurgitation and an enlarged ascending aorta. The abnormal findings were related to pregnancy outcome, including autopsy results, karyotyping results, and late fetal and neonatal echocardiography. RESULTS: Cardiac abnormalities were present in 61 fetuses overall (22.2%); including structural cardiac defects in 37 fetuses (13.5%) and functional abnormalities in 24 fetuses (8.7%). Structural cardiac abnormalities were associated with abnormal karyotype in 24 fetuses and normal karyotype in 13 fetuses. Of the 24 fetuses with functional cardiac abnormalities, 2 (8.3%) had isolated tricuspid regurgitation and 22 (91.7%) had enlarged ascending aorta. Abnormal karyotype was present in this group in 4 cases (16.7%). CONCLUSION: Increased NT can be used to define a high-risk group that should receive specialized early fetal echocardiography. This is a reliable technique with great potential for the diagnosis of both structural and functional cardiac abnormalities.

Small-bowel tumors diagnosed by wireless capsule endoscopy: report of five cases.

Tumors of the small bowel, both benign and malignant, are relatively uncommon. As the symptoms are vague and conventional diagnostic tests are unsatisfactory, these tumors often present a clinical, radiological, and endoscopic challenge. We report here on five patients in whom small-bowel tumors were diagnosed using wireless capsule endoscopy. The indications for capsule endoscopy were: obscure gastrointestinal bleeding in four patients (one jejunal capillary hemangioma, one ileal hemangiosarcoma, and two jejunal gastrointestinal stromal tumors) and chronic abdominal pain in one patient (ileal carcinoid). Wireless capsule endoscopy, a new endoscopic method, promises to improve the diagnosis of deep small-bowel pathology.

Detection of fetal structural abnormalities at the 11-14 week ultrasound scan.

The aim of this study was to evaluate the detection of fetal structural abnormalities by the 11-14 week scan. 2853 pregnant women were submitted to a routine ultrasound scan between the 11th and 14th week and the fetal skull, brain, spine, abdominal wall, limbs, stomach and bladder were examined. Following the scans the patients were examined in the second or third trimester of pregnancy. An isolated increased nuchal translucency was not considered an abnormality. However, these patients had an early echocardiography assessment. Fetal structural abnormalities were classified as major or minor and of early or late onset. A total of 130 (4.6%) defects were identified and 29 (22.3%) of these were diagnosed at the 11-14 week scan, including nine cardiac defects associated with increased nuchal translucency. The antenatal ultrasound detection rate was 71.5%, and 31.2% were detected in the first-trimester assessment. 78.8% of the major defects were diagnosed by the prenatal scan and 37.8% by the 11-14 week scan. Fetal structural abnormalities at the 11-14 week scan were detected in approximately 22.3% of the cases, therefore, a second-trimester anomaly scan is important in routine antenatal care to increase the prenatal detection of fetal defects.

Trisomy 10: ultrasound features and natural history after first trimester diagnosis.

We report on the ultrasound features and natural history of trisomy 10. At 12 weeks' gestation in a routine scan examination, the fetus presented with increased nuchal translucency thickness, mild skin oedema, bilateral pleural effusion, marked micrognathia, cardiomegaly, unilateral talipes and reversed A-wave in the ductus venosus blood flow. Karyotyping on chorionic villus sampling (CVS) led to the diagnosis of trisomy 10, which was confirmed by fetal blood sampling at 22 weeks' gestation. As the parents opted to continue with the pregnancy, the natural history and following ultrasound features are described. This is the third case of trisomy 10 in the literature reporting on the physical features. The most frequent ultrasound findings presented in trisomy 10 are increased nuchal translucency, micrognathia, renal agenesis, facial cleft, limbabnormalities, cardiac defects and early severe growth retardation.

[Echocardiographic diagnosis of transposition of the great arteries associated with anomalous pulmonary venous connection].

We report 2 cases of transposition of the great arteries associated with anomalous pulmonary venous connection emphasizing the clinical findings, the diagnosis, and the evolution of the association. One of the patients had the anomalous pulmonary venous connection in its total infradiaphragmatic form, in the portal system, and the other patient had a partial form, in which an anomalous connection of the left superior lobar vein with the innominate vein existed. At the time of hospital admission, the patients had cyanosis and respiratory distress with clinical findings suggesting transposition of the great arteries. The diagnosis in 1 of the cases, in which the anomalous connection was partial, was established only with echocardiography, without invasive procedures that would represent risk for the patient; in the other case, in which the anomalous connection was total, the malformation was only evidenced with catheterization. The patients underwent surgery for anatomical correction of the heart disease. Only 1 patient had a good outcome.

Double-outlet left ventricle. Echocardiographic diagnosis.

This is a case report of a double-outlet left ventricle associated with tricuspid atresia and hypoplasia of the right ventricle, diagnosed during echocardiography with color-flow imaging, in a three-month-old child who presented with fatigue and cyanosis. The child underwent palliative pulmonary arterial banding without an invasive procedure, and showed sustained improvement during follow-up.

Inhibition of T-cell response by Escherichia coli heat-labile enterotoxin-treated epithelial cells.

Escherichia coli heat-labile enterotoxin (LT) is an extensively studied adjuvant of mucosal responses. Nevertheless, its mode of action as an adjuvant remains incompletely understood. In this study, we describe a simplified in vitro model with which to look at some aspects of immunoregulation by LT. The interaction of LT with the apical surface of a monolayer of CaCo-2 epithelial cells induces the release of a soluble factor which inhibits the antigen-induced release of interleukin-2 by T cells cultured at the basolateral side of the cells. The release of this factor requires the ADP-ribosylating activity of LT since the isolated B subunit, as well as an enzymatically silent LT mutant, loses biological activity in this model. The inhibitory activity is likely to be due to prostaglandin release, since it is blocked by indomethacin. The contribution of LT-induced prostaglandin release to the complex immunoregulatory activity of LT is discussed.

World experience of percutaneous ultrasound-guided balloon valvuloplasty in human fetuses with severe aortic valve obstruction.

Prenatal alleviation of severe fetal aortic valve obstructions by percutaneous ultrasound-guided balloon valvuloplasty has been performed to improve the fate of affected fetuses. The purpose of this study was to analyze the current world experience of these procedures in human fetuses. Data from 12 human fetuses were available for analysis. The mean gestational age at intervention was 29.2 weeks (range 27 to 33). The mean time period between initial presentation and intervention was 3.3 weeks (range 3 days to 9 weeks). Technically successful balloon valvuloplasties were achieved in 7 fetuses, none of whom had an atretic valve. Only 1 of these fetuses remains alive today. Of the 5 remaining technical failures, 1 patient with severe aortic stenosis underwent successful postnatal intervention and remains alive. Six patients who survived prenatal intervention died from cardiac dysfunction or at surgery in the first days or weeks after delivery. Four fetuses died early within 24 hours after the procedure, 1 from a bleeding complication, 2 from persistent bradycardias, and 1 at valvotomy after emergency delivery. Thus, the early clinical experience of percutaneous ultrasound-guided fetal balloon valvuloplasty in human fetuses with severe aortic valve obstruction has been poor due to selection of severe cases, technical problems during the procedure, and high postnatal operative mortality in fetuses who survived gestation. Improved patient selection and technical modifications in interventional methods may hold promise to improve outcome in future cases.