Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831)

Abstract: The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs) was as follows: CD34 (positive), CD14 (negative), CD45 (negative), CD73 (positive), CD79 (negative), CD90 (positive), CD105 (positive), demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.

Chemical and volatile composition of jujube wines fermented by Saccharomyces cerevisiae with and without pulp contact and protease treatment

Abstract This study evaluated the chemical and volatile composition of jujube wines fermented with Saccharomyces cerevisiae A1.25 with and without pulp contact and protease treatment during fermentation. Yeast cell population, total reducing sugar and methanol contents had significant differences between nonextracted and extracted wine. The nonextracted wines had significantly higher concentrations of ethyl 9-hexadecenoate, ethyl palmitate and ethyl oleate than the extracted wines. Pulp contact also could enhance phenylethyl alcohol, furfuryl alcohol, ethyl palmitat and ethyl oleate. Furthermore, protease treatment can accelerate the release of fusel oils. The first principal component separated the wine from the extracted juice without protease from other samples based on the higher concentrations of medium-chain fatty acids and medium-chain ethyl esters. Sensory evaluation showed pulp contact and protease could improve the intensity and complexity of wine aroma due to the increase of the assimilable nitrogen.

Climate strategies of firms in the automotive and pulp & paper industries in Brazil: insights from an international perspective

This article focuses on the results of the final stage of research into the climate strategies of firms in the automotive and pulp-and-paper industries in Brazil, a country that is becoming increasingly important also in terms of climate change issues. In the first stage, the Climate Strategy Model (CSM) was developed to assess whether firms were adopting the necessary practices to assure the successful implementation of climate strategies. In the second, the CSM was applied to firms in the above mentioned industries that were chosen because of their important role in the domestic economy. In the final stage, interviews with executives of these firms were conducted to identify root causes of climate strategy implementation deficiencies and obtain new insights from an international perspective.

Internal consistency of the self-reporting questionnaire-20 in occupational groups

ABSTRACT OBJECTIVE To assess the internal consistency of the measurements of the Self-Reporting Questionnaire (SRQ-20) in different occupational groups. METHODS A validation study was conducted with data from four surveys with groups of workers, using similar methods. A total of 9,959 workers were studied. In all surveys, the common mental disorders were assessed via SRQ-20. The internal consistency considered the items belonging to dimensions extracted by tetrachoric factor analysis for each study. Item homogeneity assessment compared estimates of Cronbach’s alpha (KD-20), the alpha applied to a tetrachoric correlation matrix and stratified Cronbach’s alpha. RESULTS The SRQ-20 dimensions showed adequate values, considering the reference parameters. The internal consistency of the instrument items, assessed by stratified Cronbach’s alpha, was high (> 0.80) in the four studies. CONCLUSIONS The SRQ-20 showed good internal consistency in the professional categories evaluated. However, there is still a need for studies using alternative methods and additional information able to refine the accuracy of latent variable measurement instruments, as in the case of common mental disorders.

Growth curves of Leishmania braziliensis braziliensis Promastigotes and surface antigen expression before and after adaptation to Schneider's drosophila medium as assessed by anti-Leishmania human sera

Leishmania braziliensis braziliensis(MHOM/BR/75/M2903) was grown in Schneider's Drosophila medium. In one set of experiments promastigotes were already adapted to the medium by means of serial passages whereas in the second cells were grown in a biphasic medium and transfered to the liquid. Growth was more abundant for culture medium adapted cells; degenerate cells in small numbers as well as dead ones were present from day 5 for promastigotes adapted to liquid medium and from day 3 for newly adapted cells. Synthesis of surface antigens differed according to length of cell culture as assessed by the titer of five mucocutaneous leishmaniasis sera on subsequent days. Five days of culture for cells already adapted to the culture medium and 3 days for newly adapted ones were judged to be the best for the preparation of immunofluorescence antigens.

Induction of iron regulated proteins during normal growth of Neisseria meningitidis in a chemically defined medium

The expression of iron regulated proteins (IRPs) in vitro has been obtained in the past by adding iron chelators to the culture after bacterial growth, in the presence of an organic iron source. We have investigated aspects concerning full expression of the meningococcal IRPs during normal growth, in defined conditions using Catlin medium, Mueller Hinton and Tryptic Soy Broth (TSB). The expression of IRPs varied between different strains with respect to Ethylenediamine Di-ortho-Hidroxy-phenyl-acetic acid (EDDA) concentrations, and according to culture medium, and also between different lots of TSB. For each strain, a specific set of IRPs were expressed and higher EDDA concentrations, or addition of glucose, or use of different culture media did not resulted in a differential expression of IRPs. We were not able to grow N. meningitidis under normal growth conditions using Desferal. We looked for a good yield of outer membrane vesicles (OMVs) expressing IRPs in iron-deficient Catlin medium containing EDDA and Hemin. Culture for 32 h at 30ºC after growing for 16 h at 37ºC supported good bacterial growth. Bacterial lysis was noted after additional 24 h at 30ºC. Approximately 4 times more OMVs was recoverable from a culture supernatant after 24 h at 30ºC than from the cells after 16 h at 37ºC. The IRP were as well expressed in OMVs from culture supernatant obtained after 24 h at 30ºC as from the cells after 16 h at 37ºC.

Use of selective medium for Burkholderia cepacia isolation in respiratory samples from cystic fibrosis patients

Burkholderia cepacia colonizes cystic fibrosis (CF) patients. We evaluated the impact of the use of a selective medium in the rate of B. cepacia recovery from respiratory samples of CF patients. During a 6-month period, respiratory samples were collected from 106 CF patients and cultivated on selective media including a B. cepacia selective medium. Confirmation of the identity of B. cepacia isolates was carried out by species specific PCR and determination of genomovar status performed by a sequential PCR approach. Results of B. cepacia isolation during this period were compared to the preceding two years, when the sample processing was identical except for the lack of the B. cepacia selective medium. B. cepacia was isolated in 11/257 (4.2%) of the samples using the selective medium, in contrast with the preceding two years, when it was isolated in 6/1029 samples (0.58%), p < 0.0001. Identity of all 11 isolates was confirmed by PCR and genomovar determination was accomplished in all but one isolate. These results suggest that the use of a selective medium increases recovery rate of B. cepacia from respiratory samples.

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.

CO-INFECTION OF DENGUE VIRUS BY SEROTYPES 1 AND 4 IN PATIENT FROM MEDIUM SIZED CITY FROM BRAZIL

SUMMARY The natural co-infection with dengue virus can occur in highly endemic areas where different serotypes have been observed for many years. We report one case of DENV-1/DENV-4 co-infection in human serum detected by molecular tests. Phylogenetic analysis of the sequences obtained indicated the presence of genotype V and II for DENV-1 and DENV-4, respectively.

The comparison of the Butzler medium, filtration technique and their association with isolation of Campylobacter ssp

Campylobacter was isolated in 178 out of 622 stool samples (200 porcine, 220 bovine, and 202 canine). From these 178 samples, the microorganism was identified in 64 samples (36%) isolated only in Butzler selective medium (BSM), 34 samples (19%) using filtration technique (FT), and in 80 samples (45%) using both BSM and FT. Comparison of the proportion of positivity using both techniques showed a significant value (c2 = 9,184; p > 0.001); BSM (36%) being more efficient than FT (19%). The use of both techniques yielded the highest isolation positivity (45%).

Medium-term cryopreservation of rabies virus samples

Introduction The cryopreservation of rabies virus has been described in detail in the literature. To date, little information is available on the use of cryoprotective agents for cold preservation of this virus, and the available data focus only on short-term virus preservation. In this study, we investigated the medium-term cryopreservation of samples of rabies virus using different cryopreservation protocols. Methods The cryopreservation protocols for the rabies virus samples were performed at -20°C and were divided according to the variables of time and cryoprotectant type used. The laboratory tests (intracerebral inoculation of mice, viral titration and direct immunofluorescence) were performed at regular intervals (360 and 720 days) to assess the viability of the viral samples according to the different preservation techniques used. Results After 1 year of cryopreservation, the fluorescence intensity of intracellular corpuscles of the rabies virus and the median survival time of the mice differed between the positive controls and the treatments with the cryoprotectants. After 2 years, most of the samples subjected to the cryopreservation protocols (including the controls) did not produce fluorescence. However, the virus samples exposed to the cryoprotectant sucrose (68% solution) responded positively in the direct immunofluorescence assay and in the intracerebral inoculation of the mice. Conclusions Medium-term cryopreservation of the rabies virus inactivates the viral sample. However, the cryoprotectant agent sucrose (68%) produces a preservative effect in cryopreserved rabies virus samples.

Flowering levels, harvest season and yields of cupuassu (Theobroma grandiflorum)

Cupuassu (Theobroma grandiflorum), a specie native to Amazonia, has been planted commercially in Brazil to satisfy the demand for the flavorful juice obtained from the pulp around its seeds. The trees are notorious for low and irregular fruit production. Data gathered over two seasons from trees in a germplasm collection in Pará, Brazil, showed that some of them fruited more regularly than others. Differences in fruit production correlated to differences in flower production. Tree-to-tree variation in flower production, fruit production, and consistency of both over time suggest considerable scope for improving yields by selection. Hand pollinations resulted in a much higher frequency of fruit set than open pollinations, indicating that lack of effective pollination is also a reason for low yield. However, attempts to increase the level of effective pollination are handicapped by low knowledge about the pollinators of cupuassu and their behavior.

The Portuguese language version of social phobia and Anxiety Inventory: analysis of items and internal consistency in a Brazilian sample of 1,014 undergraduate students

OBJECTIVE: Theoretical and empirical analysis of items and internal consistency of the Portuguese-language version of Social Phobia and Anxiety Inventory (SPAI-Portuguese). METHODS: Social phobia experts conducted a 45-item content analysis of the SPAI-Portuguese administered to a sample of 1,014 university students. Item discrimination was evaluated by Student's t test; interitem, mean and item-to-total correlations, by Pearson coefficient; reliability was estimated by Cronbach's alpha. RESULTS: There was 100% agreement among experts concerning the 45 items. On the SPAI-Portuguese 43 items were discriminative (p < 0.05). A few inter-item correlations between both subscales were below 0.2. The mean inter-item correlations were: 0.41 on social phobia subscale; 0.32 on agoraphobia subscale and 0.32 on the SPAI-Portuguese. Item-to-total correlations were all higher then 0.3 (p < 0.001). Cronbach's alphas were: 0.95 on the SPAI-Portuguese; 0.96 on social phobia subscale; 0.85 on agoraphobia subscale. CONCLUSION: The 45-item content analysis revealed appropriateness concerning the underlying construct of the SPAI-Portuguese (social phobia, agoraphobia) with good discriminative capacity on 43 items. The mean inter-item correlations and reliability coefficients demonstrated the SPAI-Portuguese and subscales internal consistency and multidimensionality. No item was suppressed in the SPAI-Portuguese but the authors suggest that a shortened SPAI, in its different versions, could be an even more useful tool for research settings in social phobia.

Abundance of Conepatus chinga (Carnivora, Mephitidae) and other medium-sized mammals in grasslands of southern Brazil

Between January 2007 and December 2010, the abundance of medium-sized mammals was studied, with special focus on the Molina's hog-nosed skunk, Conepatus chinga (Molina, 1782), at four locations in southern Brazil. In this study, transect line methodology was used to obtain data for Distance Analyses. Transects were traveled by car at night, searching with spotlights along the edges of secondary roads in agricultural landscapes. Along 1,811 km, we obtained 620 observations of 20 mammal species. The most common species was the exotic European hare, Lepus europaeus (Pallas, 1778); the highest abundance estimated for South America was observed in one of the study areas, where its density was estimated as 32 individuals/km². Carnivores were the most commonly recorded mammals, represented by 10 species and comprising 51% of all observations. Molina's hog-nosed skunk occurred in all study areas, but occurred in sufficient numbers to obtain density estimates in only two of the areas. We estimated 1.4 to 3.8 individuals/km², in the first density estimate made by the transect method for a member of Conepatus in the Neotropics. These values are similar to those estimated for North American species of Mephitidae. In Brazil, C. chinga is apparently more abundant in the Pampa biome than in the grasslands of the Atlantic Forest. For two other carnivores, Lycalopex gymnocercus (Fisher, 1814) and Cerdocyon thous (Linnaeus, 1766), we estimated preliminary densities that were similar to those previously cited for different regions.

Diversity and habitat preference of medium and large-sized mammals in an urban forest fragment of southwestern Amazon

We assessed the species composition and abundance of medium and large-sized mammals in an urban forest fragment in the Brazilian Amazon, and recorded the preference of some species for particular phytophysiognomies. We placed nine transects with 20 sand plots each in three phytophysiognomies: open rainforest with a dominance of bamboos (OFB), open rainforest with palm trees (OFP), and dense rainforest (DF). We calculated species abundance as the number of records/plot.day, in a total of 2,700 plots.day. We recorded twelve mammal species; Sylvilagus brasiliensis (Linnaeus, 1758) and Dasyprocta fuliginosa (Wagler, 1831) were the most abundant. The results differed among phytophysiognomies: DF presented the highest mammal diversity, whereas the species composition of OFP was less similar than that of other phytophysiognomies. Rodents showed higher preference for OFP and Sylvilagus brasiliensis was more abundant in OFB. The study area showed high species richness, with the occurrence of mesopredators, but there was a predominance of common species adaptable to disturbed environments, which reflects the severe isolation degree of the forest fragment and the hunting pressure that is still present.