Ações educativas sobre prevenção de acidentes infantis com professoras de educação infantil

Pós-graduação em Educação - FFC

Saúde bucal e qualidade de vida em indivíduos com paralisia cerebral e Síndrome de Down: percepção dos cuidadores

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fluorose dentária em Araraquara-SP: prevalência, severidade e associação com fatores de risco

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Processamento auditivo (central) em escolares das séries iniciais de alfabetização

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Community Life and Housing in Rural Nebraska: 2005 Nebraska Rural Poll Results

Many rural communities are experiencing population decline. However, rural residents have continued to show a strong attachment to their communities. How do rural Nebraskans feel about their community? Are they satisfied with the services provided? Do they own their home? What is the condition of their home? This report details 2,851 responses to the 2005 Nebraska Rural Poll, the tenth annual effort to understand rural Nebraskans’ perceptions. Respondents were asked a series of questions about their community and housing. Trends for some of these questions are examined by comparing data from the nine previous polls to this year’s results. For all questions, comparisons are made among different respondent subgroups, that is, comparisons by age, occupation, region, etc. Based on these analyses, some key findings emerged: Rural Nebraskans’ views of the change in their community are similar to those expressed last year. This year, 28 percent believe their community has changed for the better, compared to 26 percent last year. And, in 2005, only 20 percent think their community has changed for the worse, compared to 22 percent last year. The proportion of expected movers who plan to leave the state decreased this year. Last year, 56 percent of the persons planning to move from their community expected to leave the state. That proportion decreased to 47 percent this year. Rural Nebraskans living in or near the largest communities are more likely than persons living in or near the smaller communities to say their community has changed for the better. Thirty-nine percent of persons living in or near communities with populations of 10,000 or more believe their community has changed for the better during the past year, but only 15 percent of persons living in or near communities with less than 500 people share this opinion. The community services and amenities that rural Nebraskans are most dissatisfied with include: entertainment, retail shopping and restaurants. At least one-third of rural Nebraskans express dissatisfaction with these three services. They are most satisfied with parks and recreation, library services, basic medical care services, highways and bridges, and education (K - 12). At least one-half of rural Nebraskans are satisfied with the following items in their community: appearance of residential areas (66%), crime control (61%), maintenance of sidewalks and public areas (57%) and noise (54%). Rural Nebraskans generally have positive views about their community. Sixty percent agree that their community is an ideal place to live and 52 percent say their community has good business leaders. Rural Nebraskans have mixed opinions about the future of their community. Fortyfour percent agree that their community’s future looks bright, but 42 percent disagree with this statement. Fourteen percent have no opinion. Rural Nebraskans living in or near the larger communities are more likely than residents of the smaller communities to think their community’s future looks bright. Fifty-nine percent of persons living in or near communities with populations of 10,000 or more agree with this statement, compared to only 25 percent of residents living in or near communities with less than 500 people. Further, 61 percent of the residents of the smallest communities disagree with this statement, compared to only 28 percent of the residents of the largest communities. Over three-quarters of rural Nebraskans disagree that younger residents of their community tend to stay there after completing high school. Seventy-six percent disagree with this statement, 16 percent have no opinion and eight percent agree that younger residents stay after completing high school. When comparing responses by age, younger persons are more likely than older persons to agree that younger residents stay in their community after high school. Sixteen percent of persons age 19 to 29 agree with this statement, compared to only six percent of persons age 50 to 64. Younger persons are more likely than older persons to be planning to move from their community next year. Fifteen percent of persons between the ages of 19 and 29 are planning to move next year, compared to only two percent of persons age 65 and older. An additional 17 percent of the younger respondents indicate they are uncertain if they plan to move. Most rural Nebraskans own their home. Eighty-four percent of rural Nebraskans own their home. Older persons are more likely than younger persons to own their home. Eighty-eight percent of persons over the age of 50 own their home, compared to only 52 percent of persons age 19 to 29. Housing in rural Nebraska has an average age of 50 years. Twenty-four percent of residences were built before 1930. Another 24 percent were built between 1930 and 1959. Twenty-nine percent were built between 1960 and 1979 and the remaining 24 percent were built in 1980 or later. The housing stock in smaller communities is older than the housing located in larger communities. Over one-third (35%) of the residences in communities with less than 1,000 people were built before 1930. Only 12 percent of the homes in communities with populations of 10,000 or more were built in this time period. Most rural Nebraskans appear satisfied with their home. Only 24 percent say the current size of their home does not meet their needs. The same proportion (24%) say their home is in need of major repairs. Thirty-eight percent agree that their home needs a lot of routine maintenance, but 87 percent like the location (neighborhood) of their home. One-third of rural Nebraskans living in or near the smallest communities say their home is in need of major repairs. Only 19 percent of persons living in or near communities with populations of 5,000 or more are facing this problem. Home ownership is very important to most rural Nebraskans. Eighty-two percent believe it is very important to own their home. An additional 12 percent say it is somewhat important and six percent say it is not at all important. However, persons who do not currently own their home do not feel it is important for them to do so. Only 32 percent of renters say it is very important to own their home, compared to 91 percent of home owners. And, 35 percent of renters say it is not at all important to own their home.

Processamento auditivo (central) em escolares das séries iniciais de alfabetização

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Low-energy electron collisions with glycine

We report cross sections for elastic electron scattering by gas phase glycine (neutral form), obtained with the Schwinger multichannel method. The present results are the first obtained with a new implementation that combines parallelization with OpenMP directives and pseudopotentials. The position of the well known pi* shape resonance ranged from 2.3 eV to 2.8 eV depending on the polarization model and conformer. For the most stable isomer, the present result (2.4 eV) is in fair agreement with electron transmission spectroscopy assignments (1.93 +/- 0.05 eV) and available calculations. Our results also point out a shape resonance around 9.5 eV in the A' symmetry that would be weakly coupled to vibrations of the hydroxyl group. Since electron attachment to a broad and lower lying sigma* orbital located on the OH bond has been suggested the underlying mechanism leading to dissociative electron attachment at low energies, we sought for a shape resonance around similar to 4 eV. Though we obtained cross sections with the target molecule at the equilibrium geometry and with stretched OH bond lengths, least-squares fits to the calculated eigenphase sums did not point out signatures of this anion state (though, in principle, it could be hidden in the large background). The low energy (similar to 1 eV) integral cross section strongly scales as the bond length is stretched, and this could indicate a virtual state pole, since dipole supported bound states are not expected at the geometries addressed here. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.3687345]

OmpL1 Is an Extracellular Matrix- and Plasminogen-Interacting Protein of Leptospira spp.

Leptospirosis is a zoonosis with multisystem involvement caused by pathogenic strains of the genus Leptospira. OmpL1 is an outer membrane protein of Leptospira spp. that is expressed during infection. In this work, we investigated novel features of this protein. We describe that OmpL1 is a novel leptospiral extracellular matrix (ECM)-binding protein and a plasminogen (PLG) receptor. The recombinant protein was expressed in Escherichia coli BL21(DE3) Star/pLysS as inclusion bodies, refolded, and purified by metal-chelating chromatography. The protein presented a typical beta-strand secondary structure, as evaluated by circular dichroism spectroscopy. The recombinant protein reacted with antibodies in serum samples from convalescent leptospirosis patients with a high specificity compared to serum samples from individuals with unrelated diseases. These data strengthen the usefulness of OmpL1 as a diagnostic marker of leptospirosis. The characterization of the immunogenicity of recombinant OmpL1 in inoculated BALB/c mice showed that the protein has the capacity to elicit humoral and cellular immune responses, as denoted by high antibody titers and the proliferation of lymphocytes. We demonstrate that OmpL1 has the ability to mediate attachment to laminin and plasma fibronectin, with KD (equilibrium dissociation constant) values of 2,099.93 +/- 871.03 nM and 1,239.23 +/- 506.85 nM, respectively. OmpL1 is also a PLG receptor, with a KD of 368.63 +/- 121.23 nM, capable of generating enzymatically active plasmin. This is the first report that shows and characterizes OmpL1 as an ECM-interacting and a PLG-binding protein of Leptospira spp. that may play a role in bacterial pathogenesis when expressed during infection.

Oral immunization with attenuated Salmonella vaccine expressing Escherichia coli O157:H7 intimin gamma triggers both systemic and mucosal humoral immunity in mice

Human infections with EHEC such as O157:H7 have been a great concern for worldwide food-industry surveillance. This pathogen is commonly associated with bloody diarrhea that can evolve to the life-threatening hemolytic uremic syndrome. Animals are the natural reservoir where this pathogen remains asymptomatically, in steps of ingestion and colonization of the bowel. The bacterium is shed in the feces, contaminating the surroundings, including water and food that are directed for human consumption. A major player in this colonization process is intimin, an outer membrane adhesion molecule encoded by the E. coli attachment and effacement (eae) gene that has been shown to be essential for intimate bacterial attachment to eukaryotic host cells. In an attempt to reduce the colonization of animal reservoirs with EHEC O157:H7, we designed a vaccine model to induce an immune response against intimin gamma. The model is based on its recombinant expression in attenuated Salmonella, used as a suitable vaccine vector because of its recognized ability to deliver recombinant antigens and to elicit all forms of immunity: mucosal, systemic, and humoral responses. To test this model, mice were orally immunized with a S. enterica serovar Typhimurium strain carrying the pYA3137eaeA vector, and challenged with E. coli O157:H7. Here we show that immunization induced the production of high levels of specific IgG and IgA antibodies and promoted reduction in the fecal shedding of EHEC after challenge. The live recombinant vaccine reported herein may contribute to the efforts of reducing animal intestinal mucosa colonization.

Lsa30, a novel adhesin of Leptospira interrogans binds human plasminogen and the complement regulator C4bp

Pathogenic Leptospira is the etiological agent of leptospirosis, a life-threatening disease that affects populations worldwide. Surface proteins have the potential to promote several activities, including adhesion. This work aimed to study the leptospiral coding sequence (CDS) LIC11087, genome annotated as hypothetical outer membrane protein. The LIC11087 gene was cloned and expressed in Escherichia coil BL21 (DE3) strain by using the expression vector pAE. The recombinant protein tagged with N-terminal 6XHis was purified by metal-charged chromatography and characterized by circular dichroism (CD) spectroscopy. The recombinant protein has the ability to mediate attachment to the extracellular matrix (ECM) components, laminin and plasma fibronectin, and was named Lsa30 (Leptospiral surface adhesin of 30 kDa). Lsa30 binds to laminin and to plasma fibronectin in a dose-dependent and saturable manner, with dissociation equilibrium constants (K-D) of 292 +/- 24 nM and 157 +/- 35 nM, respectively. Moreover, the Lsa30 is a plasminogen (PLC) receptor, capable of generating plasmin, in the presence of activator. This protein may interfere with the complement cascade by interacting with C4bp regulator. The Lsa30 is probably a new surface protein of Leptospira as revealed by immunofluorescence assays with living organisms and the reactivity with antibodies present in serum samples of experimentally infected hamsters. Thus, Lsa30 is a novel versatile protein that may play a role in mediating adhesion and may help pathogenic Leptospira to overcome tissue barriers and to escape the immune system. (C) 2012 Elsevier Ltd. All rights reserved.

An unusual feature of yolk sac placentation in Necromys lasiurus (Rodentia, Cricetidae, Sigmodontinae)

We studied the development of the inverted yolk sac in a New World rodent, Necromys lasiurus during early placentation. Ten implantation sites were investigated by means of histology, immunohistochemistry and electron microscopy. The yolk sac was villous near its attachment to the placenta. Elsewhere it was non-villous and closely attached to the uterus. The uterine glands were shallow and wide mouthed. They were associated with vessels and filled with secretion, suggesting the release of histotroph. This feature was absent at later stages. The intimate association of the yolk sac with specialized glandular regions of the uterus may represent a derived character condition of Necromys and/or sigmodont rodents. (C) 2012 Elsevier Ltd. All rights reserved.

Involvement of TSSA (trypomastigote small surface antigen) in Trypanosoma cruzi invasion of mammalian cells

TSSA (trypomastigote small surface antigen) is a polymorphic mucin-like molecule displayed on the surface of Trypanosoma cruzi trypomastigote forms. To evaluate its functional properties, we undertook comparative biochemical and genetic approaches on isoforms present in parasite stocks from extant evolutionary lineages (CL Brener and Sylvio X-10). We show that CL Brener TSSA, but not the Sylvio X-10 counterpart, exhibits dose-dependent and saturable binding towards non-macrophagic cell lines. This binding triggers Ca2+-based signalling responses in the target cell while providing an anchor for the invading parasite. Accordingly, exogenous addition of either TSSA-derived peptides or specific antibodies significantly inhibits invasion of CL Brener, but not Sylvio X-10, trypomastigotes. Non-infective epimastigote forms, which do not express detectable levels of TSSA, were stably transfected with TSSA cDNA from either parasite stock. Although both transfectants produced a surface-associated mucin-like TSSA product, epimastigotes expressing CL Brener TSSA showed a similar to 2-fold increase in their attachment to mammalian cells. Overall, these findings indicate that CL Brener TSSA functions as a parasite adhesin, engaging surface receptor(s) and inducing signalling pathways on the host cell as a prerequisite for parasite internalization. More importantly, the contrasting functional features of TSSA isoforms provide one appealing mechanism underlying the differential infectivity of T. cruzi stocks.

Electron Interactions with Disulfide Bridges

Because of its electronic properties, sulfur plays a major role in a variety of metabolic processes and, more in general, in the chemistry of life. In particular, S-S bridges between cysteines are present in the amino acid backbone of proteins. Protein disulfur radical anions may decay following different paths through competing intra and intermolecular routes, including bond cleavage, disproportionation, protein-protein cross linking, and electron transfer. Indeed, mass spectrometry ECD (electron capture dissociation massspectroscopy) studies have shown that capture of low-energy (<0.2 eV) electrons by multiply protonated proteins is followed by dissociation of S-S bonds holding two peptide chains together. In view of the importance of organic sulfur chemistry, we report on electron interactions with disulphide bridges. To study these interactions we used as prototypes the molecules dimethyl sulfide [(CH3)2S] and dimethyl disulfide [(H3C)S2(CH3)]. We seek to better understand the electron-induced cleavage of the disulfide bond. To explore dissociative processes we performed electron scattering calculations with the Schwinger Multichannel Method with pseudopotentials (SMCPP), recently parallelized with OpenMP directives and optimized with subroutines for linear algebra (BLAS) and LAPACK routines. Elastic cross sections obtained for different S-S bond lengths indicate stabilization of the anion formed by electron attachment to a σ*SS antibonding orbital, such that dissociation would be expected.

Support infrastructures for multimedia services with guaranteed continuity and QoS

Advances in wireless networking and content delivery systems are enabling new challenging provisioning scenarios where a growing number of users access multimedia services, e.g., audio/video streaming, while moving among different points of attachment to the Internet, possibly with different connectivity technologies, e.g., Wi-Fi, Bluetooth, and cellular 3G. That calls for novel middlewares capable of dynamically personalizing service provisioning to the characteristics of client environments, in particular to discontinuities in wireless resource availability due to handoffs. This dissertation proposes a novel middleware solution, called MUM, that performs effective and context-aware handoff management to transparently avoid service interruptions during both horizontal and vertical handoffs. To achieve the goal, MUM exploits the full visibility of wireless connections available in client localities and their handoff implementations (handoff awareness), of service quality requirements and handoff-related quality degradations (QoS awareness), and of network topology and resources available in current/future localities (location awareness). The design and implementation of the all main MUM components along with extensive on the field trials of the realized middleware architecture confirmed the validity of the proposed full context-aware handoff management approach. In particular, the reported experimental results demonstrate that MUM can effectively maintain service continuity for a wide range of different multimedia services by exploiting handoff prediction mechanisms, adaptive buffering and pre-fetching techniques, and proactive re-addressing/re-binding.

Molecular basis oh herpes simplex virus entry into the cell and retargeting of the viral tropism for the design of oncolytic herpesviruses

Herpes simplex virus 1 (HSV-1) infects oral epitelial cells, then spreads to the nerve endings and estabilishes latency in sensory ganglia, from where it may, or may not reactivate. Diseases caused by virus reactivation include mild diseases such as muco-cutaneous lesions, and more severe, and even life-threatening encephalitis, or systemic infections affecting diverse organs. Herpes simplex virus represents the most comprehensive example of virus receptor interaction in Herpesviridae family, and the prototype virus encoding multipartite entry genes. In fact, it encodes 11-12 glycoproteins and a number of additional membrane proteins: five of these proteins play key roles in virus entry into subsceptible cells. Thus, glycoprotein B (gB) and glycoprotein C (gC) interact with heparan sulfate proteoglycan to enable initial attachment to cell surfaces. In the next step, in the entry cascade, gD binds a specific surface receptor such as nectin1 or HVEM. The interaction of glycoprotein D with the receptor alters the conformation of gD to enable the activation of gB, glycoprotein H, and glycoprotein L, a trio of glycoproteins that execute the fusion of the viral envelope with the plasma membrane. In this thesis, I described two distinct projects: I. The retargeting of viral tropism for the design of oncolytic Herpesviruses: • capable of infecting cells through the human epitelial growth factor receptor 2 (HER2), overexpressed in highly malignant mammary and ovarian tumors and correlates with a poor prognosis; • detargeted from its natural receptors, HVEM and nectin1. To this end, we inserted a ligand to HER2 in gD. Because HER2 has no natural ligand, the selected ligand was a single chain antibody (scFv) derived from MAb4D5 (monoclonal antibody to HER2), herein designated scHER2. All recombinant viruses were targeted to HER2 receptor, but only two viruses (R-LM113 and R-LM249) were completely detargeted from HVEM and nectin1. To engineer R-LM113, we removed a large portion at the N-terminus of gD (from aa 6 to aa 38) and inserted scHER2 sequence plus 9-aa serine-glycine flexible linker at position 39. On the other hand, to engineer R-LM249, we replaced the Ig-folded core of gD (from aa 61 to aa 218) with scHER2 flanked by Ser-Gly linkers. In summary, these results provide evidence that: i. gD can tolerate an insert almost as big as gD itself; ii. the Ig-like domain of gD can be removed; iii. the large portion at the N-terminus of gD (from aa 6 to aa 38) can be removed without loss of key function; iv. R-LM113 and R-LM249 recombinants are ready to be assayed in animal models of mammary and ovary tumour. This finding and the avaibility of a large number of scFv greatly increase the collection of potential receptors to which HSV can be redirected. II. The production and purification of recombinant truncated form of the heterodimer gHgL. We cloned a stable insect cell line expressing a soluble form of gH in complex with gL under the control of a metalloprotein inducible promoter and purified the heterodimer by means of ONE-STrEP-tag system by IBA. With respect to biological function, the purified heterodimer is capable: • of reacting to antibodies that recognize conformation dependent epitopes and neutralize virion infectivity; • of binding a variety cells at cell surface. No doubt, the availability of biological active purified gHgL heterodimer, in sufficient quantities, will speed up the efforts to solve its crystal structure and makes it feasible to identify more clearly whether gHgL has a cellular partner, and what is the role of this interaction on virus entry.