Healing outcomes at implants installed in grafted sites: an experimental study in dogs

Aim: To evaluate the integration of implants installed using a surgical guide in augmented sites with autologous bone or deproteinized bovine bone mineral (DBBM) blocks, concomitantly with a collagen membrane.Material and methods: Mandibular molars were extracted bilaterally in six Labrador dogs, the buccal bony wall was removed, and a box-shaped defect was created. After 3 months, flaps were elevated, a bony graft was harvested from the ascending ramus, and secured to the lateral wall of the defect by means of screws. In the left mandibular side, a DBBM block was fixed into the defect. A resorbable membrane was applied at both sides, and the flaps were sutured. After 3 months, flaps were elevated, and a customized device was used as surgical guide to prepare the recipient sites in the interface between grafts and parent bone. One implant was installed in each side of the mandible. After 3 months, biopsies were harvested, and ground sections were prepared for histologic evaluation.Results: One autologous bone block graft was lost before implant installation. The width of the alveolar crest at the test sites (DBBM) was 5.4 +/- 1.2 mm before, 9.4 +/- 1.2 mm immediately after grafting, and 9.3 +/- 1 mm at implant installation. At the control sites (autologous bone), the corresponding values were: 5.2 +/- 1, 9 +/- 1.2, and 8.7 +/- 0.9 mm, respectively. All implants installed were available for histologic evaluation (n = 5). The autologous bone grafts, rich in vessels and cells, were integrated in the parent bone, and only little non-vital bone was found. The BIC% was 56.7 +/- 15.6% and 54.2 +/- 13.2% at the buccal and lingual aspects, respectively. At the test sites, the DBBM appeared to be embedded into connective tissue, and very little newly formed bone was encountered within the grafts. The BIC% was 5.8 +/- 12.3% and 51.3 +/- 14.2% at the buccal and lingual aspects, respectively.Conclusions: Autologous bone blocks used to augment the alveolar bony crest horizontally allowed the complete osseointegration of implants installed after 3 months of healing. However, similar blocks of DBBM did not promote osseointegration, although the installed implants were stable owing to the osseointegration in the sites of the parent bone.

Optical emission and electron capture of rare-earth trivalent ions located at distinct sites in SnO(2) thin films

We present photoluminescence and decay of photo excited conductivity data for sol-gel SnO(2) thin films doped with rare earth ions Eu(3+) and Er(3+), a material with nanoscopic crystallites. Photoluminescence spectra are obtained under excitation with several monochromatic light sources, such as Kr(+) and Ar(+) lasers, Xe lamp plus a selective monochromator with UV grating, and the fourth harmonic of a Nd: YAG laser (4.65eV), which assures band-to-band transition and energy transfer to the ion located at matrix sites, substitutional to Sn(4+). The luminescence structure is rather different depending on the location of the rare-earth doping, at lattice symmetric sites or segregated at grain boundary layer, where it is placed in asymmetric sites. The decay of photo-excited conductivity also shows different trapping rate depending on the rare-earth concentration. For Er-doped films, above the saturation limit, the evaluated capture energy is higher than for films with concentration below the limit, in good agreement with the different behaviour obtained from luminescence data. For Eu-doped films, the difference between capture energy and grain boundary barrier is not so evident, even though the luminescence spectra are rather distinct.

Western blot analysis of tick antigens from a Rhipicephalus sanguineus unfed larval extract and identification of antigenic sites in tick sections using immunohistochemistry. A comparative study between resistant and susceptible host species

Most parasite-host relationships are characterized by the development of resistance by the host, thus limiting the number of parasites. However, some cases are very unusual. In the relationship of the domestic dog with the brown dog-tick Rhipicephalus sanguineus this does not occur, whereas guinea pigs develop efficient resistance. Sera from domestic dogs, crab-eating foxes and guinea pigs collected before and after infestation with R. sanguineus ticks, and after immunization with a whole tick adult or larval homogenate, were used in Western blot analysis to compare and identify potential important antigens from a tick larval homogenate. The same sera were tested in an indirect immunohistochemistry assay in an attempt to compare relevant antigenic sites on histological tick sections. The immunoblotting displayed antigens recognized only by the guinea pigs, as well as several shared antigens between host species, depending on the kind of immunization. Immunohistochemistry revealed probable antigenic sites on the cells and tissues of ticks, which varied depending on the kind of immunization (infestation or vaccination) and the animal species involved.

Glucose-induced insulin secretion is impaired and insulin-induced phosphorylation of the insulin receptor and insulin receptor substrate-1 are increased in protein-deficient rats

Malnutrition is related to diabetes in tropical countries. In experimental animals, protein deficiency may affect insulin secretion. However, the effect of malnutrition on insulin receptor phosphorylation and further intracellular signaling events is not known. Therefore, we decided to evaluate the rate of insulin secretion and the early molecular steps of insulin action in insulin-sensitive tissues of an animal model of protein deficiency. Pancreatic islets isolated from rats fed a standard (17%) or a low (6%) protein diet were studied for their secretory response to increasing concentrations of glucose in the culture medium. Basal as well as maximal rates of insulin secretion were significantly lower in the islets isolated from rats fed a low protein diet. Moreover, the dose-response curve to glucose was significantly shifted to the right in the islets from malnourished rats compared with islets from control rats. During an oral glucose tolerance test, there were significantly lower circulating concentrations of insulin in the serum of rats fed a low protein diet in spite of no difference in serum glucose concentration between the groups, suggesting an increased peripheral insulin sensitivity. Immunoblotting and immunoprecipitation were used to study the phosphorylation of the insulin receptor and the insulin receptor substrate-1 as well as the insulin receptor substrate-1-p85 subunit of phosphatidylinositol 3-kinase association in response to insulin. Values were greater in hind-limb muscle from rats fed a low protein diet compared with controls. No differences were detected in the total amount of protein corresponding to the insulin receptor or insulin receptor substrate-1 between muscle from rats fed the two diets. Therefore, we conclude that a decreased glucose-induced insulin secretion in pancreatic islets from protein-malnourished rats is responsible, at least in part, for an increased phosphorylation of the insulin receptor, insulin receptor substrate-1 and its association with phosphatidylinositol 3-kinase. These might represent some of the factors influencing the equilibrium in glucose concentrations observed in animal models of malnutrition and undernourished subjects.

The use of shallow seismic reflection technique in near surface exploration of urban sites: An evaluation in the city of SãO Paulo, Brazil

In order to evaluate the use of shallow seismic technique to delineate geological and geotechnical features up to 40 meters depth in noisy urban areas covered with asphalt pavement, five survey lines were conducted in the metropolitan area of São Paulo City. The data were acquired using a 24-bit, 24-channel seismograph, 30 and 100 Hz geophones and a sledgehammer-plate system as seismic source. Seismic reflection data were recorded using a CMP (common mid point) acquisition method. The processing routine consisted of: prestack band-pass filtering (90-250 Hz); automatic gain control (AGC); muting (digital zeroin) of dead/noisy traces, ground roll, air-wave and refracted-wave; CMP sorting; velocity analyses; normal move-out corrections; residual static corrections; f-k filtering; CMP stacking. The near surface is geologically characterized by unconsolidated fill materials and Quaternary sediments with organic material overlying Tertiary sediments with the water table 2 to 5 m below the surface. The basement is composed of granite and gneiss. Reflections were observed from 40 milliseconds to 65 ms two-way traveltime and were related to the silt clay layer and fine sand layer contact of the Tertiary sediments and to the weathered basement. The CMP seismic-reflection technique has been shown to be useful for mapping the sedimentary layers and the bedrock of the São Paulo sedimentary basin for the purposes of shallow investigations related to engineering problems. In spite of the strong cultural noise observed in these urban areas and problems with planting geophones we verified that, with the proper equipment, field parameters and particularly great care in data collection and processing, we can overcome the adverse field conditions and to image reflections from layers as shallow as 20 meters.

Spectroscopic characteristics of Er3+ in the two crystallographic sites of Gd2SiO5

Gd2SiO5 is among the interesting and suitable hosts for Er3+ which find extensive applications in the infrared, visible and ultraviolet spectral regions. In order to investigate its potentialities, a prior study of the spectroscopic behaviour of Er3+ substituting for Gd3+ ions in the two crystallographic sites of the host was performed. Absorption, excitation, site-selective emission and time-resolved spectroscopies were employed in the visible spectral region to study transitions between excited 4S3/2 and ground 4I15/2 states. These levels multiplets were attributed to each site separately, and their corresponding 4S3/2 lifetimes (1.8 ± 0.1 μs for site 1 and 3.2 ± 0.1 μs for site 2) were determined.

Distinct regions of loss of heterozygosity on 22q at different sites of head and neck squamous cell carcinomas

Background: Frequent loss of heterozygosity (LOH) has been reported in many types of cancer, including head and neck carcinomas. Somatic deletions involving specific chromosomal regions are strongly associated with inactivation of the allele of a tumor suppressor gene located within the deleted region. In most studies concerning LOH in head and neck squamous cell carcinomas (HNSCC) the different anatomical sites are not distinguished. The behavior of tumors arising at various sites differs significantly, however, suggesting different intrinsic tumor properties. In this study we compared the LOH on 22q and its relationship to clinicopathological parameters at the three major sites of HNSCC: oral cavity, larynx and pharynx. Material/Methods: LOH and microsatellite instability (MSI) were studied using seven polymorphic microsatellite markers mapped to the 22q11-q13.3 region in 37 oral, 32 laryngeal, and 31 pharyngeal carcinomas. Results: Two separate regions of LOH were identified in the laryngeal (22q11.2-12.1) and oral cavity (22q13.1-13.31) tumors. When the different anatomical sites were compared, a statistically significant difference was found between the presence of LOH at D22S421 (p<0.001), D22S315 (p=0.014) and D22S929 (p=0.026) in the laryngeal tumors. Conclusions: These data suggest that distinct regions on 22q are involved in LOH in oral cavity and laryngeal tumorigenesis but do not support a similar association between the development of pharyngeal tumors and genes located on 22q. These findings implicate the presence of different tumor suppressor genes mapping to distinct regions on chromosome 22q in oral and laryngeal carcinomas.

Sensitivity of AMPA receptor channel to calcium oscillations: A computational study

We used a computational model of biochemical pathways that are involved in the phosphorylation/dephosphorylation of AMPA receptor to study the receptor responses to calcium oscillations. In the model, the biochemical pathways are assumed to be located immediately under the postsynaptic membrane and we included three states of AMPA receptor: dephosphorylated, and phosphorylated in one or in two sites. To characterize the effects of calcium oscillations on the AMPA receptor, we exposed the model to stimuli with three varying parameters, namely frequency, number of pulses and calcium spike duration. Our model showed sensitivity to all of these three parameters. © 2002 Elsevier Science B.V. All rights reserved.

Mapping eIF5A binding sites for Dys1 and Lia1: In vivo evidence for regulation of eIF5A hypusination

The evolutionarily conserved factor eIF5A is the only protein known to undergo hypusination, a unique posttranslational modification triggered by deoxyhypusine synthase (Dys1). Although eIF5A is essential for cell viability, the function of this putative translation initiation factor is still obscure. To identify eIF5A-binding proteins that could clarify its function, we screened a two-hybrid library and identified two eIF-5A partners in S. cerevisiae: Dys1 and the protein encoded by the gene YJR070C, named Lia1 (Ligand of eIF5A). The interactions were confirmed by GST pulldown. Mapping binding sites for these proteins revealed that both eIF5A domains can bind to Dys1, whereas the C-terminal domain is sufficient to bind Lia1. We demonstrate for the first time in vivo that the N-terminal α-helix of Dys1 can modulate enzyme activity by inhibiting eIF5A interaction. We suggest that this inhibition be abrogated in the cell when hypusinated and functional eIF5A is required. © 2003 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

Physiological and cellular responses in two populations of the mussel Perna perna collected at different sites from the Coast of São Paulo, Brazil

The physiological conditions of mussels from Ubatuba and Santos and also of organisms transplanted from Ubatuba to Santos were studied by using different techniques. Assays for lysosomal stability were conducted on the haemolymph. Heart rate activity was monitored for 6h. The embryonic development of larvae obtained from the collected mussels was analysed. For all the compared groups of mussels, no significant differences were observed for the cardiac activity monitoring and the embryonic bioassays. The mean Neutral Red (NR) retention time was similar for the animals from Santos and Ubatuba, whereas the organisms transplanted to Santos showed a reduction in the retention time of the dye, indicating damage in the lysosomal membranes. These differences were possibly due to environmental factors, but further investigations are required to confirm this hypothesis.

Eucalyptus ESTs corresponding to the enzyme glutamine synthetase and the protein D1, sites of action of herbicides that cause oxidative stress

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Localization of rDNA sites in holocentric chromosomes of three species of triatomines (Heteroptera, Triatominae).

Chromatin organization in the holocentric chromosomes of three triatomines species was cytologically studied by fluorescent in situ hybridization with a 45S rDNA probe of Drosophila melanogaster to localize ribosomal genes. In Triatoma tibiamaculata, metaphases I showed telomeric highlights in a single, larger bivalent. In T. protacta, hybridization was detected in one of the telomeres of an autosomal chromosome. In T. platensis, there were highlights in a single, smaller chromosome (X chromosome). The results obtained did not agree with the expected localization of rDNA genes in the sex chromosomes of triatomines, as demonstrated by silver impregnation, and suggest that the chromosome reorganization that occurred in this group during evolution may be a more important mechanism involved in rDNA distribution.

Calling sites and acoustic partitioning in species of the Hyla nana and rubicundula groups (Anura, Hylidae)

We analysed spatial and acoustic partitioning among four species of Hyla belonging to two species-groups: nana (H. nana and H. sanborni) and rubicundula (H. elianeae and H. jimi). Field activities were conducted at three permanent ponds, from 1998 through 2001. Four attributes of the calling sites were analysed: perch height, distance of the perch from the edge of the pond, type of perch (vegetation) and the individual's position on the perch. There was extensive overlap in the four calling-site variables analysed. However, we found spatial segregation did occur in calling site height and the distance of perches from pond edges. Bioacoustic analyses revealed behavioural differences among species in calling activity, both time of onset and peak calling in chorus. There was acoustic partitioning among species the fundamental frequency of the advertisement calls, principally as a function of the temporal structure (e.g. note duration, rate of note repetition, duration and rate of repetition of the calling pulses). We propose that differences in physical attributes of calling site and in characteristics of calls allow these species to exist in sympatry.