A distinct tymovirus infecting Cassia hoffmannseggii in Brazil

Leaves of Cassia hoffmannseggii, a wild fabaceous species found in the Atlantic Forest, with a severe mosaic symptom were collected in Pernambuco State, Brazil. By transmission electron microscopy, two types of virus particles were found: the first was recognized as particles of a potyvirus, which was later identified as Cowpea aphid-borne mosaic virus; and the second was isometric and present in high concentration. The observation of vesicles at the periphery of chloroplasts suggested a tymovirus infection, which was confirmed by subsequent assays. A serological assay against several tymovirus antisera resulted in positive reaction of this tymo-like virus with an antiserum of Passion fruit yellow mosaic virus. By means of RT-PCR and using degenerated primers for the conserved region of RNA-dependent RNA polymerase (RdRp) gene of tymoviruses, a specific DNA fragment was amplified and sequenced. Based on this sequence, a specific forward primer was synthesized and successfully used to amplify the 3' terminal genome region, containing the partial RdRp gene and the complete coat protein (CP) sequences. The CP was 188 amino acids (aa) long, and the highest CP aa identity was observed with Kennedya yellow mosaic virus (61 %). Based on the current ICTV demarcation criterion, this isolate was considered as a distinct tymovirus and tentatively named as Cassia yellow mosaic-associated virus.

A retrospective serologic survey of hantavirus infections in the county of Cassia dos Coqueiros, state of Sao Paulo, Brazil

Introduction: In recent years, hantavirus infections producing severe diseases have obtained an increased attention from public health authorities from the countries of Eurasia to the Americas. Brazil has reported 1,300 cases of hantavirus cardiopulmonary syndrome (HCPS) from 1993 to 2010, with about 80 of them occurring in the northeast of the state of Sao Paulo, with 48% fatality rate. Araraquara virus was the causative agent of HCPS in the region. Considering that hantaviruses causing human disease in the Americas were unknown until 1993, we have looked for hantavirus infections in the population of Cassia dos Coqueiros county, northeast of the state of Sao Paulo, Brazil, before this time. This county has about 2,800 inhabitants and an economy based on agriculture, including cultivation of Brachiaria decumbens grass. The grass seeds are an important rodent attraction, facilitating transmission of hantavirus to man. Four HCPS cases were reported so far in the county. Methods: In this study, 1,876 sera collected from 1987 to 1990 were tested for IgG to hantavirus by IgG-ELISA, using the N recombinant protein of Araraquara virus as antigen. Results: Positive results were observed in 89 (4.7%) samples, which were all collected in 1987. The positivity among urban inhabitants was 5.3%, compared with 4.3% among those living in rural areas. Conclusions: Our results showed that hantavirus infections occurred in Cassia dos Coqueiros, completely unrecognized, even before hantaviruses were described in the Americas.

BRAZILIAN OFFSHORE WAVE CLIMATE BASED ON NWW3 REANALYSIS

This paper provides a description of the wave climate off the Brazilian coast based on an eleven-year time series (Jan/1997-Dec/2007) obtained from the NWW3 operational model hindcast reanalysis. Information about wave climate in Brazilian waters is very scarce and mainly based on occasional short-term observations, the present analysis being the first covering such temporal and spatial scales. To define the wave climate, six sectors were defined and analyzed along the Brazilian shelf-break: South (W1), Southeast (W2), Central (W3), East (W4), Northeast (W5) and North (W6). W1, W2 and W3 wave regimes are determined by the South Atlantic High (SAH) and the passage of synoptic cold fronts; W4, W5 and W6 are controlled by the Intertropical Convergence Zone (ITCZ) and its meridional oscillation. The most energetic waves are from the S, generated by the strong winds associated to the passage of cold fronts, which mainly affect the southern region. Wave power presents a decrease in energy levels from south to north, with its annual variation showing that the winter months are the most energetic in W1 to W4, while in W5 and W6 the most energetic conditions occur during the austral summer. The information presented here provides boundary conditions for studies related to coastal processes, fundamental for a better understanding of the Brazilian coastal zone.

ICHTHYOPLANKTON DISTRIBUTION AND ABUNDANCE IN THE NORTHERN TODOS OS SANTOS AND CAMAMU BAYS, BAHIA STATE - BRAZIL

The occurrence, distribution and abundance of ichthyoplankton in Todos os Santos and Camamu Bays were analyzed based on four samplings (winter 2003, summer 2003, winter 2004 and summer 2005). Samples were obtained by surface horizontal hauls, using a 200-mu m mesh conical-cylinder plankton net. The distribution and abundance of eggs indicate a remarkable seasonal and annual variation of spawning activity in the region, especially when the two summer campaigns are compared. In summer 2003 the highest quantitative values were recorded, especially for Camamu, where the maximum reached 106.56 eggs.m(-3), with an overall average of 43.46 eggs.m(-3) for the two areas. In summer 2005 values were relatively low, the overall average being 3.49 eggs.m(-3). The larval taxonomic composition is characterized by the predominance of gobiids, with small variation from summer to winter. Considering all the campaigns and samplings undertaken in both areas, larvae of 11 families were identified: Engraulidae, Clupeidae, Mugilidae, Atherinopsidae, Hemiramphidae, Syngnathidae, Blenniidae, Carangidae, Gobiidae, Achiridae and Tetraodontidae.

Anthropogenic influences in a lagoonal environment: a multiproxy approach at the valo grande mouth, Cananéia-Iguape system (SE Brazil)

The Cananeia-Iguape system, SE Brazil, consists of a complex of lagoonal channels, located in a United Nations Educational, Scientific and Cultural Organization (UNESCO) Biosphere Reserve. Nevertheless, important environmental changes have occurred in approximately the last 150 yrs due to the opening of an artificial channel, the Valo Grande, connecting the Ribeira de Iguape River to the lagoonal system. Our objective is to assess the historical record of the uppermost layers of the sedimentary column of the lagoonal system in order to determine the history of environmental changes caused by the opening of the artificial channel. In this sense, an integrated geochemical-faunal approach is used. The environmental changes led significant modifications in salinity, in changes of the depositional patterns of sediments and foraminiferal assemblages (including periods of defaunation), and, more drastically, in the input of heavy metals to the coastal environment. The concentrations Pb in the core analyzed here were up to two times higher than the values measured in contaminated sediments from the Santos estuary, the most industrialized coastal zone in Brazil.

Purification and characterization of aprotinin from porcine lungs

Aprotinin, the most studied serine proteinase inhibitor, was isolated from porcine lung for the first time. The purified porcine aprotinin had an Mr value of similar to 7 kDa. It cross- reacted with polyclonal serum anti- commercial aprotinin. About 1 mu g porcine aprotinin inhibited 6 mu g trypsin whereas 1 mu g commercial soybean inhibitor inhibited only 1 mu g trypsin. The aprotinin gene was also isolated from porcine lung: the deduced amino acid sequence showed 74% identity to bovine aprotinin.

Body temperature, Aldrete-Kroulik index, and patient discharge from the Post-Anesthetic Recovery Unit

Patient discharge from post-anesthetic recovery (PAR) depends, among other factors, on normothermia and the patient's score on the Aldrete-Kroulik index. The objective of this study was to verify the relationship between the Aldrete-Kroulik index and body temperature in patients. This study was performed at the University of Sao Paulo University Hospital. Convenience sampling was used, and the sample consisted of 60 patients of ages between 18 and 60 years who underwent general anesthesia. The patients' body temperature was obtained by tympanic measurement, and the Aldrete-Kroulik index was measured on admission and at discharge from post-anesthetic recovery. The data were processed using SPSS, considering a significance level of 5%, and the Spearman and Wilcoxon tests were applied. In conclusion, no significant correlation was found between the two parameters for discharge.

Influence of the use of Aliquat 336 in the immobilization procedure in sol-gel of lipase from Bacillus sp ITP-001

Aliquat 336, a liquid hydrophobic material, was used at different concentrations (0.5-3.0%, w/v) as an additive in the preparation of encapsulated lipase from Bacillus sp. ITP-001 on sol-gel silica matrices using tetraethoxysilane (TEOS) as the precursor. The resulting hydrophobic matrices and immobilized lipases were characterized with regard to specific surface area (BET method), adsorption-desorption isotherms, pore volume (Vp) and size (dp) by nitrogen adsorption (BJH method) and scanning electron microscopy (SEM). The catalytic activities and the corresponding coupling yields were assayed in the hydrolysis of olive oil. In comparison with pure silica matrices, the immobilization process in the presence of Aliquat 336 decreased the values for specific surface area and increased the values for pore specific volume (Vp) and mean pore diameter (dp). This behavior may be related to the partial adsorption of the enzyme on the external surface of the hydrophobic matrix as indicated by scanning electron microscopy. Aliquat 336 concentrations in the range from 0.5 to 1.5% (w/v) provided immobilized derivatives with higher coupling yields and better substrate affinity. The highest coupling yield (Y-A = 71%) was obtained for the immobilized enzyme prepared in the presence of 1.5% Aliquat which gave the following morphological properties: specific surface area = 183 m(2)/g, pore specific volume (Vp) = 0.36 cc/g and mean pore diameter (dp)= 91 angstrom. (c) 2012 Elsevier B.V. All rights reserved.

Gram-negative osteomyelitis: clinical and microbiological profile

Introduction: Despite the growing interest in the study of Gram-negative bacilli (GNB) infections, very little information on osteomyelitis caused by GNB is available in the medical literature. Objectives and methods: To assess clinical and microbiological features of 101 cases of osteomyelitis caused by GNB alone, between January 2007 and January 2009, in a reference center for the treatment of high complexity traumas in the city of Sao Paulo. Results: Most patients were men (63%), with median age of 42 years, affected by chronic osteomyelitis (43%) or acute osteomyelitis associated to open fractures (32%), the majority on the lower limbs (71%). The patients were treated with antibiotics as inpatients for 40 days (median) and for 99 days (median) in outpatient settings. After 6 months follow-up, the clinical remission rate was around 60%, relapse 19%, amputation 7%, and death 5%. Nine percent of cases were lost to follow-up. A total of 121 GNB was isolated from 101 clinical samples. The most frequently isolated pathogens were Enterobacter sp. (25%), Acinetobacter baumannii (21%) e Pseudomonas aeruginosa (20%). Susceptibility to carbapenems was about 100% for Enterobacter sp., 75% for Pseudomonas aeruginosa and 60% for Acinetobacter baumannii. Conclusion: Osteomyelitis caused by GNB remains a serious therapeutic challenge, especially when associated to nonfermenting bacteria. We emphasize the need to consider these agents in diagnosed cases of osteomyelitis, so that an ideal antimicrobial treatment can be administered since the very beginning of the therapy. (C) 2012 Elsevier Editora Ltda. All rights reserved.

Assessing Pathogenicity for Novel Mutation/Sequence Variants: The Value of Healthy Older Individuals

Improvement in DNA technology is increasingly revealing unexpected/unknown mutations in healthy persons and generating anxiety due to their still unknown health consequences. We report a 44-year-old healthy father of a 10-year-old daughter with bilateral coloboma and hearing loss, but without muscle weakness, in whom a whole-genome CGH revealed a deletion of exons 38-44 in the dystrophin gene. This mutation was inherited from her asymptomatic father, who was further clinically and molecularly evaluated for prognosis and genetic counseling (GC). This deletion was never identified by us in 982 Duchenne/Becker patients. To assess whether the present case represents a rare case of non-penetrance, and aiming to obtain more information for prognosis and GC, we suggested that healthy older relatives submit their DNA for analysis, to which several complied. Mutation analysis revealed that his mother, brother, and 56-year-old maternal uncle also carry the 38-44 deletion, suggesting it an unlikely cause of muscle weakness. Genome sequencing will disclose mutations and variants whose health impact are still unknown, raising important problems in interpreting results, defining prognosis, and discussing GC. We suggest that, in addition to family history, keeping the DNA of older relatives could be very informative, in particular for those interested in having their genome sequenced.

Common chromosomal imbalances and stemness-related protein expression markers in endometriotic lesions from different anatomical sites: the potential role of stem cells

Endometriosis is a multifactorial gynecological disease characterized by the presence of functional endometrium-like tissue in ectopic sites. Several studies have focused on elucidating the immunological, endocrine, environmental and genetic factors involved in endometriosis. However, its pathogenesis is still unclear. High-resolution comparative genomic hybridization was applied to screen for genomic imbalances in laser microdissected stromal and epithelial cells from 20 endometriotic lesions and three samples of eutopic endometrium derived from eight patients. The expression of seven stemness-related markers (CD9, CD13, CD24, CD34, CD133, CD117/c-Kit and Oct-4) in endometrial tissue samples was evaluated by immunohistochemistry. Samples of eutopic endometrium showed normal genomic profiles. In ectopic tissues, an average of 68 genomic imbalances was detected per sample. DNA losses were more frequently detected and involved mainly 3p, 5q, 7p, 9p, 11q, 16q, 18q and 19q. Many of the genomic imbalances detected were common to endometriotic stroma and epithelia and also among different endometriotic sites from the same patient. These findings suggested a clonal origin of the endometriotic cells and the putative involvement of stem cells. Positive immunostaining for CD9, CD34, c-Kit and Oct-4 markers was detected in isolated epithelial and/or stromal cells in eutopic and ectopic endometrium in the majority of cases. The presence of shared genomic alterations in stromal and epithelial cells from different anatomical sites of the same patient and the expression of stemness-related markers suggested that endometriosis arises as a clonal proliferation with the putative involvement of stem cells.

Functional Limitation and Intermittent Claudication: Impact of Blood Pressure Measurements

Background: Arterial hypertension is an important risk factor for Lower-Limb Occlusive Arterial Disease (LLOAD). However, the correlation between blood pressure and pulse pressure (PP) with LLOAD severity and functional impairment resulting from this disease is not well established in the Brazilian population. Objective: To verify whether there is a correlation between blood pressure, PP, LLOAD severity and functional capacity in patients with symptomatic LLOAD. Methods: A total of 65 patients (62.2 + 8.1 years, 56.9% males) were evaluated. They were divided into two groups: normal (A) and high (B) blood pressure. LLOAD severity was assessed using the ankle-brachial index (ABI) and functional capacity by the total and pain-free walking distance at the 6-minute walking test (6MWT). Results: Group A consisted of 17 (26.1%) patients. The systolic (SBP), diastolic blood pressure (DBP), and PP were, respectively, 125.4 +/- 11.7, 74.5 +/- 9.1 and 50.9 +/- 10.0 mmHg in group A and 160.7 +/- 19.6, 90.0 +/- 12.2 and 70.7 +/- 20.2 mmHg in group B. The ABI was significantly lower in group B (0.66 +/- 0.12 vs. 0.57 +/- 0.13, p < 0.05). SBP and PP correlated with LLOAD severity and the distances walked at the 6MWT. Patients with PP > 40 mmHg walked shorter distances. Conclusion: SBP and PP significantly correlated with the distances walked in the 6MWT, suggesting they are clinical markers of functional capacity impairment in patients with symptomatic LLOAD. (Arq Bras Cardiol 2012; 98(2): 161-166)

Nutrient Intake of Women 3 Years After Roux-en-Y Gastric Bypass Surgery

Nutritional deficiencies, especially micronutrient deficiencies, can occur in obese individuals. Surgical treatment may aggravate or cause these deficiencies, depending on the type of procedure, food intake and the use of multivitamins, minerals or other supplements. The objective of the study was to evaluate the nutrient intake of women who had undergone Roux-en-Y gastric bypass (RYGB) surgery. A cross-sectional, controlled study was conducted among 44 women after RYGB (operated-group, OG; mean years post-operation = 3.4) and a control group of 38 healthy women (non-operated group, NOG) matched by age and economic condition. The women reported their dietary intake using a 4-day record. The Dietary Reference Intakes was used as a reference. The macronutrient contributions to dietary energy intake presented an acceptable distribution for proteins and carbohydrates. Lipid intake was high among women in the OG and the NOG (43.2 and 55.3 %, respectively). In the evaluation of micronutrients, a statistically significant difference was observed between the groups for iron, zinc and vitamins B1 and B12. Both groups were at high risk for inadequate calcium intake, and the OG was at risk for inadequate zinc, iron and vitamin B1 intake. The nutrient intake of women who had undergone RYGB is very similar to that of non-operated women, with the exception of a reduced intake of iron, zinc and vitamins B1 and B12, which may be due to the difficulty of consuming meat and a balanced diet. The findings of this study emphasize the importance of appropriate nutritional intervention and the regular use of multivitamin and mineral supplements for these patients.

Methane emission from soil under long-term no-till cropping systems

Methane (CH4) emission from agricultural soils increases dramatically as a result of deleterious effect of soil disturbance and nitrogen fertilization on methanotrophic organisms; however, few studies have attempted to evaluate the potential of long-term conservation management systems to mitigate CH4 emissions in tropical and subtropical soils. This study aimed to evaluate the long-term effect (>19 years) of no-till grass- and legume-based cropping systems on annual soil CH4 fluxes in a formerly degraded Acrisol in Southern Brazil. Air sampling was carried out using static chambers and CH4 analysis by gas chromatography. Analysis of historical data set of the experiment evidenced a remarkable effect of high C- and N-input cropping systems on the improvement of biological, chemical, and physical characteristics of this no-tilled soil. Soil CH4 fluxes, which represent a net balance between consumption (-) and production (+) of CH4 in soil, varied from -40 +/- 2 to +62 +/- 78 mu g C m(-2) h(-1). Mean weighted contents of ammonium (NH4+-N) and dissolved organic carbon (DOC) in soil had a positive relationship with accumulated soil CH4 fluxes in the post-management period (r(2) = 0.95, p = 0.05), suggesting an additive effect of these nutrients in suppressing CH4 oxidation and stimulating methanogenesis, respectively, in legume-based cropping systems with high biomass input. Annual CH4 fluxes ranged from -50 +/- 610 to +994 +/- 105 g C ha(-1), which were inversely related to annual biomass-C input (r(2) = 0.99, p = 0.003), with the exception of the cropping system containing pigeon pea, a summer legume that had the highest biologically fixed N input (>300 kg ha(-1) yr(-1)). Our results evidenced a small effect of conservation management systems on decreasing CH4 emissions from soil, despite their significant effect restoring soil quality. We hypothesized that soil CH4 uptake strength has been off-set by an injurious effect of biologically fixed N in legume-based cropping systems on soil methanotrophic microbiota, and by the methanogenesis increase as a result of the O-2 depletion in niches of high biological activity in the surface layer of the no-tillage soil. (C) 2012 Elsevier B.V. All rights reserved.

Evaluation of Distinct Freezing Methods and Cryoprotectants for Human Amniotic Fluid Stem Cells Cryopreservation

Amniotic fluid (AF) was described as a potential source of mesenchymal stem cells (MSCs) for biomedicine purposes. Therefore, evaluation of alternative cryoprotectants and freezing protocols capable to maintain the viability and stemness of these cells after cooling is still needed. AF stem cells (AFSCs) were tested for different freezing methods and cryoprotectants. Cell viability, gene expression, surface markers, and plasticity were evaluated after thawing. AFSCs expressed undifferentiated genes Oct4 and Nanog; presented typical markers (CD29, CD44, CD90, and CD105) and were able to differentiate into mesenchymal lineages. All tested cryoprotectants preserved the features of AFSCs however, variations in cell viability were observed. In this concern, dimethyl sulfoxide (Me2SO) showed the best results. The freezing protocols tested did not promote significant changes in the AFSCs viability. Time programmed and nonprogrammed freezing methods could be used for successful AFSCs cryopreservation for 6 months. Although tested cryoprotectants maintained undifferentiated gene expression, typical markers, and plasticity of AFSCs, only Me2SO and glycerol presented workable viability ratios.