The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion

An increased plasma concentration of von Willebrand factor (vWF) is detected in individuals with many infectious diseases and is accepted as a marker of endothelium activation and prothrombotic condition. To determine whether ExoU, a Pseudomonas aeruginosa cytotoxin with proinflammatory activity, enhances the release of vWF, microvascular endothelial cells were infected with the ExoU-producing PA103 P. aeruginosa strain or an exoU-deficient mutant. Significantly increased vWF concentrations were detected in conditioned medium and subendothelial extracellular matrix from cultures infected with the wild-type bacteria, as determined by enzyme-linked immunoassays. PA103-infected cells also released higher concentrations of procoagulant microparticles containing increased amounts of membrane-associated vWF, as determined by flow cytometric analyses of cell culture supernatants. Both flow cytometry and confocal microscopy showed that increased amounts of vWF were associated with cytoplasmic membranes from cells infected with the ExoU-producing bacteria. PA103-infected cultures exposed to platelet suspensions exhibited increased percentages of cells with platelet adhesion. Because no modulation of the vWF mRNA levels was detected by reverse transcription-polymerase chain reaction assays in PA103-infected cells, ExoU is likely to have induced the release of vWF from cytoplasmic stores rather than vWF gene transcription. Such release is likely to modify the thromboresistance of microvascular endothelial cells.

Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Infection in a rat model reactivates attenuated virulence after long-term axenic culture of Acanthamoeba spp

Prolonged culturing of many microorganisms leads to the loss of virulence and a reduction of their infective capacity. However, little is known about the changes in the pathogenic strains of Acanthamoeba after long culture periods. Our study evaluated the effect of prolonged culturing on the invasiveness of different isolates of Acanthamoeba in an in vivo rat model. ATCC strains of Acanthamoeba, isolates from the environment and clinical cases were evaluated. The in vivo model was effective in establishing the infection and differentiating the pathogenicity of the isolates and re-isolates. The amoebae cultured in the laboratory for long periods were less virulent than those that were recently isolated, confirming the importance of passing Acanthamoeba strains in animal models.

Acute dacryocystitis: another clinical manifestation of sporotrichosis

Sporotrichosis associated with exposure to domestic cats is hyperendemic in Rio de Janeiro, Brazil. A review of the clinical records at our institute revealed four patients with clinical signs of dacryocystitis and a positive conjunctival culture for Sporothrix who were diagnosed with Sporothrix dacryocystitis. Three patients were children (< 13 years of age) and one patient was an adult. Two patients reported contact with a cat that had sporotrichosis. Dacryocystitis was associated with nodular, ulcerated lesions on the face of one patient and with granulomatous conjunctivitis in two patients; however, this condition manifested as an isolated disease in another patient. All of the patients were cured of the fungal infections, but three patients had chronic dacryocystitis and one patient developed a cutaneous fistula. Sporotrichosis is usually a benign disease, but may cause severe complications when the eye and the adnexa are affected. Physicians, especially ophthalmologists in endemic areas, should be aware of the ophthalmological manifestations and complications of sporotrichosis.

Knockout confirmation for Hurries: rapid genotype identification of Trypanosoma cruzi transfectants by polymerase chain reaction directly from liquid culture

Gene knockout is a widely used approach to evaluate loss-of-function phenotypes and it can be facilitated by the incorporation of a DNA cassette having a drug-selectable marker. Confirmation of the correct knockout cassette insertion is an important step in gene removal validation and has generally been performed by polymerase chain reaction (PCR) assays following a time-consuming DNA extraction step. Here, we show a rapid procedure for the identification of Trypanosoma cruzi transfectants by PCR directly from liquid culture - without prior DNA extraction. This simple approach enabled us to generate PCR amplifications from different cultures varying from 106-108 cells/mL. We also show that it is possible to combine different primer pairs in a multiplex detection reaction and even to achieve knockout confirmation with an extremely simple interpretation of a real-time PCR result. Using the “culture PCR” approach, we show for the first time that we can assess different DNA sequence combinations by PCR directly from liquid culture, saving time in several tasks for T. cruzi genotype interrogation.

Culture of mouse peritoneal macrophages with mouse serum induces lipid bodies that associate with the parasitophorous vacuole and decrease their microbicidal capacity against Toxoplasma gondii

Lipid bodies [lipid droplets (LBs)] are lipid-rich organelles involved in lipid metabolism, signalling and inflammation. Recent findings suggest a role for LBs in host response to infection; however, the potential functions of this organelle in Toxoplasma gondii infection and how it alters macrophage microbicidal capacity during infection are not well understood. Here, we investigated the role of host LBs in T. gondii infection in mouse peritoneal macrophages in vitro. Macrophages cultured with mouse serum (MS) had higher numbers of LBs than those cultured in foetal bovine serum and can function as a model to study the role of LBs during intracellular pathogen infection. LBs were found in association with the parasitophorous vacuole, suggesting that T. gondii may benefit from this lipid source. Moreover, increased numbers of macrophage LBs correlated with high prostaglandin E2 (PGE2) production and decreased nitric oxide (NO) synthesis. Accordingly, LB-enriched macrophages cultured with MS were less efficient at controlling T. gondii growth. Treatment of macrophages cultured with MS with indomethacin, an inhibitor of PGE2 production, increased the microbicidal capacity against T. gondii. Collectively, these results suggest that culture with MS caused a decrease in microbicidal activity of macrophages against T. gondii by increasing PGE2 while lowering NO production.

The organizational culture of a Brazilian public hospital

The objective of this research was to analyze the organizational culture of a Brazilian public hospital. It is a descriptive study with quantitative approach of data, developed in a public hospital of São Paulo State, Brazil. The sample was composed by 52 nurses and 146 nursing technicians and auxiliaries. Data were collected from January to June 2011 using the Brazilian Instrument for Assessing Organizational Culture – IBACO. The analysis of the organizational values showed the existence of hierarchical rigidity and centralization of power within the institution, as well as individualism and competition, which hinders teamwork. The values concerning workers’ well-being, satisfaction and motivation were not highly valued. In regard to organizational practices, the promotion of interpersonal relationship, continuous education, and rewarding practices were not valued either. It becomes apparent that traditional models of work organization support work practices and determine the organizational culture of the hospital.


SPECTRA OF MOTHERS OF PREMATURE CHILDREN ABOUT THE EDUCATIVE CIRCLE OF CULTURE

Abstract We sought to know the spectra of mothers of premature children regarding their experience with circle of culture of educational character and identifying the learning provided by the circle of culture about newborn care after hospital discharge. A descriptive study was performed in a hospital located in Fortaleza, Brazil. Three meetings of a circle of culture with 17 mothers of premature newborns were performed. The interpretation of the corpus was performed using thematic analysis. Emerged from the categories: Maternal experience in a circle of culture; Promoted social support among mothers through the circle of culture; and Learning provided by the circle of culture. It was concluded that teaching parents during the hospitalization of the child should be held in a way to involve parents in the care of the newborn, provide moments of health education, opportunities for support and dialogue between professionals and family.

Aeration condition of a clayey oxisol under long-term no-tillage

The hypothesis of this study was that the absence of soil tillage in long-term no-tillage (NT) systems can be detrimental to soil aeration. The objective was to assess the aeration condition of an Oxisol (Rhodic Ferrasol), very clayey texture (750 g kg-1 of clay; 200 g kg-1 of sand), after 30 years of cultivation under NT. The physical property soil air permeability (Ka) is sensitive to changes in the soil pore system. Aside from Ka, the air-filled porosity (ε a) and indices of pore continuity (K1 and N), derived from the relationship between Ka and εa, were used as indices of soil aeration. From the soil layers 0.0-0.1 and 0.1-0.2 m, 240 undisturbed samples were collected along a transect perpendicular to the crop rows, at three sampling positions: corn plant row (CR); center of the interrow (INT); and the equidistant point between CR and INT (PE). The properties Ka and εa were determined at soil matric potentials (Ψm) of -2, -4, -6, -10, -30, and -50 kPa. Soil bulk density (BD) was also determined. The results confirmed the hypothesis. In the 0.0-0.1 m layer, Ka, K1, N and Ψa were significantly greater and BD significantly lower in CR than at the other sampling positions. At a Ψm of -10 kPa, the Ka of CR was 6.9 and 8.4 times higher than in PE and INT, respectively, in the 0.0-0.1 m layer. The properties Ka, K1 and N were sensitive enough to detect changes in the pore system and their differences between the sampling positions demonstrated the importance of the spatial location for soil sampling. Tilling the crop rows provides better soil aeration under NT.

The new culture of electronic publishing

Local conditions in the past often limited opportunities for scholarly exchange. But now these limits are gone and the global workplace has replaced them. It is important to react to these changes. Every academic department must now adopt new methods and rethink processes. Another is the intense national and international debate about open access to scholarly knowledge. The Open Access Initiative shows that a change is taking place in the communication process. This change is also important for service departments within research institutions. Libraries, computer centers and related units have to ask themselves how to react appropriately to the new conditions. What services must be changed or redeveloped, and in what quality and quantity should they be offered? This article focuses on changes in the scholarly publication process. It describes both technological changes and the changes needed in people's attitudes.

Radiation and energy balance of lettuce culture inside a polyethylene greenhouse

The objective of this paper was to describe the radiation and energy balance, during the lettuce (Lactuca sativa, L. cv. Verônica) crop cycle inside a polyethylene greenhouse. The radiation and energy balance was made inside a tunnel greenhouse with polyethylene cover (100 mum) and in an external area, both areas with 35 m². Global, reflected and net radiation, soil heat flux and air temperature (dry and humid) were measured during the crop cycle. A Datalogger, which operated at 1 Hz frequency, storing 5 minutes averages was utilized. The global (K¯) and reflected (K­) radiations showed that the average transmission of global radiation (K¯in / K¯ex) was almost constant, near to 79.59%, while the average ratio of reflected radiation (K­in / K­ex) was 69.21% with 8.47% standard-deviation. The normalized curves of short-wave net radiation, in relation to the global radiation (K*/ K¯), found for both environments, were almost constant at the beginning of cycle; this relation decreased in the final stage of culture. The normalized relation (Rn/ K¯) was bigger in the external area, about 12%, when the green culture covered the soil surface. The long-wave radiation balance average (L*) was bigger outside, about 50%. The energy balance, estimated in terms of vertical fluxes, showed that, for the external area, in average, 83.07% of total net radiation was converted in latent heat evaporation (LE), and 18% in soil heat flux (G), and 9.96% in sensible heat (H), while inside of the greenhouse, 58.71% of total net radiation was converted in LE, 42.68% in H, and 28.79% in G.

ANATOMICAL STUDIES OF IN VITRO ORGANOGENESIS INDUCED IN LEAF-DERIVED EXPLANTS OF PASSIONFRUIT

With the aim of studying the organogenesis in vitro in Passiflora edulis Sims f. flavicarpa Deg., the passionfruit, leaf-derived explants were cultured on media containing NAA or BAP and incubated either in continuous darkness or in light. The histological events leading to de novo organ formation were evaluated. Darkness induces rhizogenesis in the presence of NAA, whereas direct shoot regeneration is stimulated by light and BAP. This latter condition is recommended for passionfruit micropropagation as several adventitious shoot buds were formed from meristemoids of parenchymal origin.

Body condition score to predict the postpartum fertility of crossbred beef cows

The relationship between changes in body condition score (BCS) during the postpartum and fertility in beef cows suckling calves under extensive conditions were investigated. Cows were subjected to four BCS evaluations over the postpartum period, starting around one month after calving. In the second evaluation cows were treated with medroxy-progesterone acetate impregnated pessaries and received an injection of estradiol benzoate. At the third evaluation, pessaries were removed and calves were separated from the cows for 96 hours, during which time estrous was observed twice a day, and animals artificially inseminated 12 hours after detection. When calves returned to their dams, bulls were introduced until a 60-day mating period was reached. The distribution of BCS differed among calving groups and evaluations. Results indicated that only cows comprising a BCS 3 (1 to 5 scale) around the first month postpartum can be used in an artificial insemination program with possibilities of becoming pregnant. There was no statistical difference between the calving groups in pregnancy rate. The evolution of the BCS of the cows during postpartum can be used to adjust the start of the breeding season to coincide with the time of the year where herd pregnancy rates will be highest.

Tissue culture parameters in sweet orange cultivars

The objective of this work was to establish tissue culture parameters for gene transfer in sweet orange cultivars. Epicotyl explants with different ages were cultured with 6-benzylaminopurine (BAP), kanamycin and hygromycin. Shoots were cultured with alpha-naphthaleneacetic acid (NAA) alone or in combination with indole-3-butyric acid (IBA). The requirement of BAP for shoot development was genotype-specific. Epicotyl explants from 35-day-old seedlings produced significantly more shoots per explant in 'Pêra'. Kanamycin inhibited shoot regeneration for the most cultivars. The percentage of shoots that produced roots in 'Pêra' was significantly higher in medium with NAA and IBA than with NAA alone.

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.