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The aim of this study was to evaluate the physicochemical characteristics of 3 mineralized bovine inorganic biomaterials and correlate them with the dissolution rate. Bio-Oss(r), GenoxInorgânico(r), and Bonefill(r) were examined using field emission gun scanning electron microscopy (FEG-SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), specific surface area (BET), calcium/phosphorous molar ratio and a dissolution assay. Bio-Oss(r) showed a micro- and nanoporous structure consisting of 15-nm hydroxyapatite (HA) crystallites; Genox(r), a microporous structure composed of 39-nm HA crystallites; and Bonefill(r), micro- and nanoporous structure of indeterminable crystallite size. FTIR analysis showed that Bio-Oss(r) and Genox(r) were composed of calcium phosphate. The absorption bands of phosphate were poorly defined in Bonefill(r). By XRD, Bio-Oss(r) was shown to contain peaks related to the carbonated HA, whereas Genox(r) only contained peaks corresponding to HA. The broad bands in Bonefill(r) indicated low crystallinity. Bio-Oss(r) showed a greater surface area and calcium release rate than that of Genox(r). Although all biomaterials were of bovine origin, the different manufacturing processes result in materials with different physicochemical properties and may influence the biological and clinical response.

Purpose: The objective of this systematic review and meta-analysis was to assess and compare the marginal bone loss around implants supporting single fixed prostheses and multiple-unit screw-retained prostheses., Materials and Methods: The literature was searched manually and electronically to identify studies in which the marginal peri-implant bone loss around single-implant prostheses and screw-retained multiple-implant prostheses was evaluated radiographically. Two reviewers independently selected the literature and extracted the data. The random-effects method was used to obtain estimates of marginal peri-implant bone loss (means and 95% confidence intervals [CIs])., Results: Of the 2,107 studies identified by a preliminary search, 17 fulfilled the inclusion criteria; 7 were related to single-implant prostheses and 10 to multiple-implant screw-retained fixed prostheses. The mean marginal peri-implant bone loss was 0.9 mm (95% CI, 0.49 to 1.32 mm) for multiple-implant screw-retained prostheses and 0.58 mm (95% CI, 0.37 to 0.80 mm) for single-implant prostheses., Conclusion: This indirect comparison provided no evidence to support the assertion that there are differences in marginal peri-implant bone loss between single implant prostheses and multiple screw-retained prostheses.

Objective: The aim of this work was to compare the effects of estradiol and genistein treatment on hyaluronic acid (HA) concentration in postmenopausal facial skin., Methods: In this study, 30 postmenopausal women were evaluated in a prospective, randomized, double-blind trial. The volunteers were postmenopausal women treated in the Gynecology Department of the Federal University of São Paulo. The participants were divided into two groups: group E, treated with 0.01% 17β-estradiol gel (n = 15), and group G, treated with 4% genistein gel (isoflavones, n = 15). The treatment lasted for 24 consecutive weeks. Preauricular skin biopsies were performed for each participant at baseline (E1 and G1) and after treatment (E2 and G2) to evaluate HA concentration in tissue. The materials were processed using immunohistochemical and biochemical methods., Results: After 24 weeks of treatment, HA concentration increased in both groups, but the effect was greater for estradiol treatment than for genistein treatment., Conclusions: Our data suggest that both treatments may enhance HA concentration in postmenopausal skin but that estrogen produces results that are greater than those produced by isoflavones.

Purpose: The objective of the present study was to realize an indirect morphometric evaluation of bone thickness lateral to the mandibular canal., Material and Methods: In 30 partially or totally edentulous dry jaws, the first and second molar areas were analyzed using computed tomography., Results: The findings indicated that 28.33% of the sites could be considered for the installation of a standard 3.75-mm-diameter implant laterally between the mandibular canal and its bony counterpart., Conclusion: Implant installation in the posterior mandible region, lateral to the mandibular canal, is an alternative to more complex techniques, such as vertical ridge augmentation.

A better understanding of the molecular mechanisms of renal cell carcinogenesis could contribute to a decrease in the mortality rate of this disease. The aim of this study was to evaluate the glycosaminoglycans profile and heparanase expression in renal cell carcinoma. The study included 24 patients submitted to nephrectomy with confirmed pathological diagnosis of renal cell carcinoma. The majority of the samples (87.5%) were classified in the initial stage of renal cell carcinoma (clinical stages I and II). Heparanase messenger ribonucleic acid expression was evaluated by quantitative real-time reverse transcription polymerase chain reaction, and sulfated glycosaminoglycans were identified and quantified by agarose gel electrophoresis of renal cell carcinoma samples or non-neoplastic tissues obtained from the same patients (control group). The sulfated glycosaminoglycans and hyaluronic acid were analyzed in urine samples of the patients before and after surgery. The data showed a significant statistical increase in chondroitin sulfate, and a decrease in heparan sulfate and dermatan sulfate present in neoplastic tissues compared with non-neoplastic tissues. Higher heparanase messenger ribonucleic acid expression in the neoplastic tissues was also shown, compared with the non-neoplastic tissues. The urine glycosaminoglycans profile showed no significant difference between renal cell carcinoma and control samples. Extracellular matrix changes observed in the present study clarify that heparanase is possibly involved with heparan sulfate turnover, and that heparanase and the glycosaminoglycans can modulate initial events of renal cell carcinoma development., (© 2012 The Japanese Urological Association.)

Objectives: To evaluate the effectiveness of a high-pressure sodium bicarbonate spray protocol to decontaminate implant surfaces intentionally inoculated with bacteria., Materials and Methods: Twenty commercially pure titanium implants, 10 with machined surfaces and 10 with rough surfaces, were inoculated with Streptococcus sanguis and then submitted to a decontamination protocol using a high-pressure sodium bicarbonate spray device for 1 minute under aseptic conditions., Results: After the application of the decontamination protocol, all bacterial cells were removed from the tested implants, regardless of surface roughness., Conclusion: The results suggest that regardless of the implant surface roughness, the protocol using high-pressure sodium bicarbonate spray for 1 minute, under aseptic conditions, was effective in removing all the viable bacterial cells.

Purpose: An in vitro model was developed and tested to evaluate the precision of guided implant systems. The accuracy of dental implants placed with a flapless technique was analyzed using a stereolithographic template in vitro. Differences between the virtual and actual positions of the implants were measured., Materials and Methods: Six polyurethane mandibles with artificial silicone gums were fabricated, and each was fitted with an individual computed tomography (CT) guide. Stereolithographic guides were created using computer-aided design/computer-assisted manufacturing technology and virtual planning software. All stereolithographic guides had four holes for stabilization pins and three holes for cylindric implants. After implant placement, the mandibles were subjected to another CT scan to compare the actual implant positions with the planned positions. The pre- and postimplantation CT images were superimposed using digital processing image software to evaluate the linear and angular deviations between the virtual planning data and the surgical results., Results: The mean angular discrepancy between the virtual and actual positions of the 18 placed implants was 2.16 ± 0.92 degrees. Among the placed implants, 66.7% were situated a mean of 0.38 ± 0.03 mm apical to the planned vertical position, and 33.3% were situated 0.39 ± 0.03 mm coronal to the planned position., Conclusions: Within the limitations of the present study, this tool showed promising accuracy in virtual implant placement.

Purpose: The aim of the present study was to verify the presence of a microgap between implants and prosthetic abutments on their external surfaces in five different external-hex implant systems and to determine whether bacterial leakage occurs through the implant-abutment (I-A) interface., Materials and Methods: Ten samples of each implant system were used. Eight samples of each implant system were inoculated with 0.3 μL of a suspension of Escherichia coli for bacterial leakage evaluation. Two other samples were used as controls. Sample analysis was accomplished at 24 and 48 hours and on the 5th, 7th, and 14th day after inoculation. After microbiologic analysis, all samples were prepared for I-A interface evaluation by scanning electron microscopy. The mean of the microgap misfit was obtained from six points at x1,000 and x3,000 magnifications., Results: The Neodent implant system (0.51 ± 0.39 μm) showed the closest adaptation, followed by Dentoflex (1.44 ± 0.73 μm), Titanium Fix (1.88 ± 1.28 μm), SIN (2.46 ± 3.38 μm), and Conexão (2.68 ± 3.02 Μm). Twenty-five percent of the Dentoflex samples showed bacterial leakage through the I-A interface; no other implant system presented bacterial leakage., Conclusion: The width of the microgap at the I-A interface was less than 3 μm in all systems. A direct correlation between bacterial leakage and I-A interface misfit was not observed.

Objective: To compare and evaluate concentrations of hyaluronic acid (HA) in normal vocal folds of both young and elderly people of both sexes., Study Design: Experimental study., Setting: The research was carried out at the ENT, Head and Neck Surgery Department and the Pharmacology Institute of São Paulo Federal University., Subjects and Methods: Enzyme-linked immunosorbent assay plates coated with biotin-conjugated binding protein and streptavidin-europium conjugate were sequentially incubated with samples from the middle portion of the cover, vocal ligament, and vocal muscle of 19 young and 20 elderly subjects of both sexes. After the release of europium in an enhancement solution, final fluorescence was measured using a fluorometer., Results: Statistically significant comparisons: (1) all subject groups combined exhibited a higher HA concentration in the cover and ligament compared with the muscle samples; (2) women subjects had a significantly lower HA concentration in muscle compared with the ligament, and male subjects showed significantly lower HA in muscle compared with the cover; and (3) both genders of young subjects showed a higher HA concentration in ligament samples compared with the muscle., Conclusion: HA concentrations are higher in the cover and ligament compared with the muscle in both genders and age groups, and there is a higher HA concentration in ligament compared with the muscle of young subjects of both genders. HA levels in the cover samples of younger women showed great variability that may relate to ovarian hormone levels, reflecting different phases of the menstrual cycle.

We studied the induction of antigen-specific IgA memory B cells (B(M)) in volunteers who received live attenuated Shigella flexneri 2a vaccines. Subjects ingested a single oral dose of 10(7), 10(8) or 10(9) CFU of S. flexneri 2a with deletions in guaBA (CVD 1204) or in guaBA, set and sen (CVD 1208). Antigen-specific serum and stool antibody responses to LPS and Ipa B were measured on days 0, 7, 14, 28 and 42. IgA B(M) cells specific to LPS, Ipa B and total IgA were assessed on days 0 and 28. We show the induction of significant LPS-specific IgA B(M) cells in anti-LPS IgA seroresponders. Positive correlations were found between anti-LPS IgA B(M) cells and anti-LPS IgA in serum and stool; IgA B(M) cell responses to IpaB were also observed. These B(M) cell responses are likely play an important role in modulating the magnitude and longevity of the humoral response., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Purpose: Because almost all implant systems reveal marginal bone loss and some implants can be affected by periimplantitis, to realize an effective maintenance treatment it is important to understand the biofilm development over different implant surfaces. The objective of this human study is to comparatively analyze the kinetics of microorganism species succession on 3 surfaces., Materials and Methods: Three commercially pure titanium sheets with different roughness were randomly fixed bilaterally on the lingual regions of Hawley-type plates of 6 volunteers. The participants used the apparatuses in the following experimental periods: 1, 3, 7, 14, and 21 days, and were instructed to remove the acrylic plates only during oral hygiene procedures. Samples of biofilm were harvested from the central areas of each sheet, and the presence of 24 bacterial species was identified using Checkerboard DNA-DNA Hybridization method., Results: The results showed no statistically significant differences with regard to the kinetics of bacterial species succession on the different surfaces, in all periods of time. Rough surfaces were not more prone to colonization by pathogenic bacteria than the smooth surfaces., Conclusion: The surface roughness does not affect the bacterial species succession in the biofilm. The succession of species was similar during all periods of the study.

Attenuated live oral typhoid vaccine candidate CVD 909 constitutively expresses Salmonella Typhi capsular polysaccharide antigen (Vi). A randomized, double-blind, heterologous prime-boost clinical study was conducted to determine whether immunity to licensed parenteral Vi vaccine could be enhanced by priming with CVD 909. Priming with CVD 909 elicited higher and persistent, albeit not significant, anti-Vi IgG and IgA following immunization with Vi, than placebo-primed recipients. Vi-specific IgA B memory (B(M)) cells were significantly increased in CVD 909-primed subjects. S. Typhi-specific LPS and flagella IgA B(M) cells were observed in subjects immunized with CVD 909 or with the licensed Vi-negative oral typhoid vaccine Ty21a. CVD 909-induced B(M) cells exhibited a classical B(M) phenotype (i.e., CD3(-)CD19(+)IgD(-)CD27(+)). This is the first demonstration of classical B(M) cells specific for bacterial polysaccharide or protein antigens following typhoid immunization. The persistent IgA B(M) responses demonstrate the capacity of oral typhoid vaccines to prime mucosally relevant immune memory., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Successful T cell priming in early postnatal life that can generate effective long-lasting responses until adulthood is critical in HIV vaccination strategies because it prevents early sexual initiation and breastfeeding transmission of HIV. A chimeric DNA vaccine encoding p55 HIV gag associated with lysosome-associated membrane protein 1 (LAMP-1; which drives the antigen to the MIIC compartment), has been used to enhance cellular and humoral antigen-specific responses in adult mice and macaques. Herein, we investigated LAMP-1/gag vaccine immunogenicity in the neonatal period in mice and its ability to generate long-lasting effects. Neonatal vaccination with chimeric LAMP/gag generated stronger Gag-specific immune responses, as measured by the breadth of the Gag peptide-specific IFN-gamma, proliferative responsiveness, cytokine production and antibody production, all of which revealed activation of CD4+ T cells as well as the generation of a more robust CTL response compared to gag vaccine alone. To induce long-lived T and B cell memory responses, it was necessary to immunize neonates with the chimeric LAMP/gag DNA vaccine. The LAMP/gag DNA vaccine strategy could be particularly useful for generating an anti-HIV immune response in the early postnatal period capable of inducing long-term immunological memory., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Liver biopsy is the gold-standard method to stage fibrosis; however, it is an invasive procedure and is potentially dangerous. The main objective of this study was to evaluate biological markers, such as cytokines IL-13, IFN-gamma, TNF-alpha and TGF-beta, platelets, bilirubins (Bil), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), total proteins, gamma-glutamil transferase (gamma-GT) and alkaline phosphatase (AP), that could be used to predict the severity of hepatic fibrosis in schistosomiasis and hepatitis C (HC) as isolated diseases or co-infections. The following patient groups were selected: HC (n = 39), HC/hepatosplenic schistosomiasis (HSS) (n = 19), HSS (n = 22) and a control group (n = 13). ANOVA and ROC curves were used for statistical analysis. P < 0.05 was considered significant. With HC patients we showed that TNF-alpha (p = 0.020) and AP (p = 0.005) could differentiate mild and severe fibrosis. With regard to necroinflammatory activity, AST (p = 0.002), gamma-GT (p = 0.034) and AP (p = 0.001) were the best markers to differentiate mild and severe activity. In HC + HSS patients, total Bil (p = 0.008) was capable of differentiating between mild and severe fibrosis. In conclusion, our study was able to suggest biological markers that are non-invasive candidates to evaluate fibrosis and necroinflammatory activity in HC and HC + HSS.

Purpose: The aim of this study was to compare the osseointegration percentage of biomimetic hydroxyapatite (HA)-coated implants on longitudinal and transverse histologic sections using scanning electron microscopy., Materials and Methods: Ten commercially pure titanium dental implants, measuring 3.75 mm in diameter and 10 mm in length, were coated with HA by the biomimetic process and installed on the proximal metaphysis of the right tibia of 10 adult New Zealand female rabbits (Oryctolagus cunniculus). After a 3-month healing period, the animals were killed and the implants were removed together with the neighboring bone tissue. The specimens were than dehydrated on a crescent concentration of ethanol solution and embedded on resin. The osseointegration percentage of biomimetic HA-coated implants on 1 longitudinal and 1 transverse section were compared, using scanning electron microscopy and computerized histomorphometry., Results: It was verified that there were no statistically significant differences between the results of the different histological section directions (P > 0.05)., Conclusion: Thus, in this study, the direction of histological sections did not affect bone-implant percentage.

Shigella dysenteriae type 1 can cause devastating pandemics with high case fatality rates; a vaccine for Shigella is unavailable currently. Because of the risks associated with performing challenge studies with wild-type S. dysenteriae 1 in human clinical trials to advance vaccine development, an improved nonhuman primate model is needed urgently. In the present study, cynomolgus macaques (Macaca fascicularis) were challenged with various doses of S. dysenteriae 1 strain 1617 to establish a dose that would produce shigellosis. Further, different routes of delivery of S. dysenteriae 1 were compared to establish the most appropriate route for infection. Animals receiving 10(11) cfu S. dysenteriae 1 intragastrically consistently developed signs of shigellosis characterized by the onset of diarrhea and dysentery within 2 to 3 d. Administration of as many as 10(9) cfu S. dysenteriae 1 intraduodenally did not elicit signs characteristic of infection in macaques despite fecal shedding of bacteria for as long as 10 d. S. dysenteriae 1 administered intraduodenally at 10(9) cfu or intragastrically at 10(11) cfu elicited robust IgG and IgA antibody responses to LPS. We have developed a reliable challenge model of infection with wild-type S. dysenteriae 1 in cynomolgus macaques that reproducibly induces disease and elicits robust immune responses. We believe that this animal model may provide unique insights into the immunologic mechanisms of protection to S. dysenteriae 1 infection and in advancing development of a vaccine against shigellosis.

Objectives: The objective of the present study is to evaluate the response to dental implants in healthy and osteoporotic bone., Materials and Methods: Ten ovariectomized (OVX) New Zealand rabbits submitted to a hypocalcic diet and 10 sham-aged rabbits were used. All animals were submitted to bone mineral density (BMD) measurements before ovariectomy, and also 4 months afterwards, using dual energy X-ray absorptiometry. The BMD measurements showed a significant loss of bone mass, between the first and second examinations, only in the experimental group (P<0.05). After the bone mass loss induction period, three different implants were installed in the proximal tibia metaphisis of each animal: a titanium alloy implant (Ti), a plasma-spray hydroxyapatite-coated implant (HA-PS), and another implant coated with hydroxyapatite with the biomimetic process (HA-B)., Results: After 3 months, histomorphometry showed a bone-to-implant contact (BIC) for Ti implants of 73.09+/-13.74% in healthy and 66.09+/-30.01% in OVX animals. The BIC for the HA-PS was 64.83+/-15.65% and 90.17+/-8.14% for healthy and OVX animals, respectively, and 88.66+/-5.30% and 87.96+/-10.71% for the HA-B implants placed in the same conditions. The differences between the implants in healthy and OVX conditions were not statistically significant (P>0.05). The only significant difference within groups was observed in the healthy animals between HA-B and Ti implants (P<0.06)., Conclusion: Within the parameters used in this animal model it was not possible to observe BIC differences between osteoporotic and healthy animals.

Investigation of the properties of two different silicate materials, namely MCM-41, with one-dimensional, cylindrical pores (4.04 nm diameter), and Na-magadiite, with a bidimensional, layered structure (1.67 nm between layers) was carried out. These materials may be used as sorbents, and their polarity is determinative to the kind of applications they may be employed at. Pyrene, which presents a vibronically resolved emission spectrum, was used as a fluorescent probe to give information on the polarity of these silicates voids. The ratio of band intensity at 383 nm (band 3) to that at 373 nm (band 1) is known to diminish as the polarity of the medium increases. Pristine sorbents and pyrene-adsorbed ones were characterized by thermogravimetric analysis and X-ray diffraction. UV-VIS emission spectra of included pyrene were recorded at temperatures ranging from 10 to 300 K, and from comparing the I(3) : I(1) ratio it was found that Na-magadiite internal sites correspond to a less polar medium than those in MCM-41. This is in accordance with the thermogravimetric data, which shows that a larger amount of pyrene was included in Na-magadiite, despite its smaller dimensions. Yet, excimer emission, usually a concentration dependent phenomenon, was mainly observed for the MCM-41 samples. Upon temperature lowering, pyrene molecules move to other sites of different polarities, towards an intermediary value for both materials.

Unlabelled: Hormone plays an important role in the larynx. Among other substances, vocal folds contain hyaluronic acid, which tissue concentration may vary according to hormone action., Aim: the objective of this study is to analyze hyaluronic acid concentration in the vocal folds during estral and gravidic-puerperal cycles., Materials and Methods: Experimental study. 40 adult rats were divided into two groups. In the first group we used 20 rats to establish the concentration of hyaluronic acid during the estral cycle and in the second group, 20 animals were submitted to the same procedure but during the gravidic-puerperal cycle., Results: Variations in hyaluronic acid concentration was not observed during the estral cycle. In the gravidic puerperal cycle group, an increase in hyaluronic acid concentration was observed in the puerperal subgroup. Comparing the two groups of estral and gravidic-puerperal cycles, no difference was observed., Conclusions: In comparing all subgroups of estral and gravidic-puerperal cycles, an increase in hyaluronic acid concentration was noticed only in the puerperal phase.

Objective: To quantify and study the immunoexpression of hyaluronic acid (HA) in the uterine horns of the mouse throughout the estrous cycle phases., Design: Experimental study using an ELISA-like fluorometric assay to quantify HA and an avidin-biotin immunoperoxidase method using biotinylated hyaluronan-binding protein for histochemical studies., Setting: University-based laboratory., Animal(s): Forty regularly cycling adult female mice were divided into four groups according to the diagnosed phase of the cycle: proestrus, estrus, metaestrus, and diestrus., Intervention(s): None., Main Outcome Measure(s): Histologic samples of the uterine horns. Immunohistochemical reaction was evaluated by detection of HA and CD44 distribution within the uterine horn. Tissue HA content was determined through an ELISA-like fluorometric assay with the same hyaluronan-binding protein and with europium-labeled streptavidin., Result(s): The immunohistochemical HA and CD44 reactions were most intense during diestrus, mainly below the luminal epithelium. HA was strongly stained in the connective tissue near the myometrium layer during metaestrus. The biochemical data showed that the highest concentration of HA in uterine horns occurred during diestrus (4053.0 +/- 651.4 ng/g dry tissue) compared with other phases., Conclusion(s): Our data show that the expression of HA in mouse uterine horns is highest during the diestrous phase. The fluctuations of HA in the mouse uterus during the estrous phase may be related to the varying estrogen and P levels during the cycle and may be important as far as embryo implantation is concerned.

Purpose: The aim of this study was to characterize implants coated with hydroxyapatite (HA) using 2 different processes., Materials and Methods: Commercially pure titanium implants were coated with HA using a modified biomimetic process and the plasma spray process. These implants were characterized by scanning electron microscopy, Fourier-transformed infrared spectroscopy, and x-ray photoelectron spectroscopy., Results: The plasma spray process generates a typical rough topography mainly consisting of HA. The HA coating produced by biomimetic process was partially dissolved in water and only a very thin layer of calcium titanate plus calcium phosphate and probably beta-tricalcium phosphate remained. In vitro test showed that both coatings can be considered bioactive., Conclusion: The modified biomimetic process seems to be a simple and low-cost alternative to coat titanium with a high potential of In vitro application.

We evaluated B memory responses in healthy adult volunteers who received one oral dose of live-attenuated Shigella flexneri 2a vaccine. LPS-specific B(M) cells increased from a median of 0 at baseline to 20 spot forming cells (SFC)/10(6) expanded cells following vaccination (p=0.008). A strong correlation was found between post-vaccination anti-LPS B(M) cell counts and peak serum anti-LPS IgG titers (rs=0.95, p=0.0003). Increases in B(M) specific for IpaB approaching significance were also observed. In sum, oral vaccination with live-attenuated S. flexneri 2a elicits B(M) cells to LPS and IpaB, suggesting that B(M) responses to Shigella antigens should be further studied as a suitable surrogate of protection in shigellosis.

Purpose: The purpose of the present study was to investigate whether osseointegration of HA-coated implants can be affected by irradiation., Materials and Methods: A single dose of 15 Gy was applied to the right tibia of 5 rabbits and the left tibia was not irradiated and used as control. After 3 months, the animals were killed and the quantification of the implant-bone contact surfaces was performed using scanning electron micrographs of the resin sections containing the implants in the irradiated and nonirradiated bones. After the healing period, all implants were stable., Results: The results show a mean percentage of bone-implant contact of 45.52% in the irradiated tissue and 47.56% in the nonirradiated tissue, with no statistic significant difference (P > 0.05)., Conclusion: In conclusion, radiotherapy may not interfere with osseointegration process around HA-coated implants in rabbit cortical bone.

Immunomics research uses in silico epitope prediction, as well as in vivo and in vitro approaches. We inoculated BALB/c (H2d) mice with 17DD yellow fever vaccine to investigate the correlations between approaches used for epitope discovery: ELISPOT assays, binding assays, and prediction software. Our results showed a good agreement between ELISPOT and binding assays, which seemed to correlate with the protein immunogenicity. PREDBALB/c prediction software partially agreed with the ELISPOT and binding assay results, but presented low specificity. The use of prediction software to exclude peptides containing no epitopes, followed by high throughput screening of the remaining peptides by ELISPOT, and the use of MHC-biding assays to characterize the MHC restrictions demonstrated to be an efficient strategy. The results allowed the characterization of 2 MHC class I and 17 class II epitopes in the envelope protein of the YF virus in BALB/c (H2d) mice.

Purpose: The surface properties of dental implants have been considered directly related to the success of the osseointegrated interface. Hydroxyapatite (HA)-coated implants promote a bioactive surface because they facilitate cellular migration and bone growth, resulting in a faster osseointegration., Materials and Methods: This study analyzed and compared 2 different implant surfaces: a group composed of titanium implants and another group coated with HA. The HA coating was produced by the biomimetic process to reduce costs., Results and Conclusions: After histomorphometric analyses it was possible to demonstrate that there were no statistically significant differences between the groups.

Objective: The purpose of this study was to evaluate the presence of headache in women with a previous history or new-onset headache during the current gestation, classify the findings, and describe the clinical characteristics and outcome of the headache., Methods: From January/1998 to June/2002 we prospectively evaluated 1101 pregnant women (12-45 years old), with a history of headache, at two prenatal clinics and an inpatient obstetric public hospital. Women were interviewed using a semi-structured questionnaire during the first, second, and third gestation trimesters and immediately after delivery. All interviews were conducted by one of the authors, using the International Headache Society Classification (IHSC-2004)., Results: In 1029 women there was a history of headache prior to the current pregnancy, 36 (3.4%) women first experienced headache during this pregnancy and 40 patients experienced new types of headache. In these 76 patients with new onset headache during pregnancy, 40 had secondary headache (52.6%), 31 had primary headache (40.8%), and 5 had headache not classified elsewhere (6.6%). According to IHSC- 2004 criteria, we found migraine in 848/1029 women (82.4%), with pregestational headache., Conclusions: Most of the pregnant women presented with headache, mainly in migraine, prior to pregnancy, and most of the headaches improved or disappeared during the second and third gestation trimester. In a relatively small number of pregnant women, a new type of headache started during the gestation.

Objective/hypothesis: To evaluate the concentration and distribution of hyaluronic acid (HA) in human adult vocal folds., Study Design: Experimental study., Methods: Histologic samples of the anterior, middle, and posterior regions of the vocal folds were studied to determine the tissue distribution of HA. The HA was traced with hyaluronan-binding proteins (HAPB) from bovine cartilage. Chemical bonds were detected through the avidin-biotin immunoperoxidase method. A control with the specific Streptomyces hyalurolyticus hyaluronidase was conducted to avoid false-positive results. The concentration levels of HA were determined through a fluorescence-based assay method with the same HAPB and europium-labeled streptavidin., Results: Women presented higher HA concentrations than men in every studied region of the vocal folds and overall HA concentration levels twice as higher. In regard to the distribution of HA, the macula flavae and the vocal fold intermediate and deep layers presented marked staining, with moderate staining around the vessels, on the gland capsules, and the connective tissue surrounding the vocal fold muscular fibers., Conclusions: HA is diffusely distributed along the vocal folds; the profuse amounts of HA observed in the female vocal folds may be related to impact absorption, acting as a protective factor against the characteristic high vibration frequency.

Purpose: To compare total protein concentration in the aqueous humor of primary open-angle glaucoma and non-glaucomatous patients., Methods: Aqueous humor samples were obtained from 22 patients just before trabeculectomy for clinically uncontrolled primary open angle glaucoma (POAG group). Aqueous humor (0.1 mL) was aspirated by inserting a 26-gauge needle into the anterior chamber. The same procedure was performed in 22 non-glaucomatous patients just before cataract surgery (control group). Immediately after collection, the aqueous humor was stored at -20 degrees C. Aqueous humor total protein concentration was determined using a colorimetric assay., Results: The geometric mean of total protein concentration of the aqueous humor samples was 32 mg/dL (range: 8-137 mg/dL) in the primary open angle glaucoma group and 16 mg/dL (range: 2-85 mg/dL) in the control group. The ratio of the protein concentration between the two groups was 2.0 (95% confidence interval: 1.3 to 3.2; p=0.003)., Conclusions: The total protein concentration in primary open-angle glaucoma aqueous humor was approximately two times higher than that in non-glaucomatous subjects.

Ag processing is a critical step in defining the repertoire of epitope-specific immune responses. In the present study, HIV-1 p55Gag Ag was synthesized as a DNA plasmid with either lysosomal-associated membrane protein-1 (LAMP/gag) or human dendritic cell-LAMP (DC-LAMP/gag) and used to immunize mice. Analysis of the cellular trafficking of these two chimeras demonstrated that both molecules colocalized with MHC class II molecules but differed in their overall trafficking to endosomal/lysosomal compartments. Following DNA immunization, both chimeras elicited potent Gag-specific T and B cell immune responses in mice but differ markedly in their IL-4 and IgG1/IgG2a responses. The DC-LAMP chimera induced a stronger Th type 1 response. ELISPOT analysis of T cell responses to 122 individual peptides encompassing the entire p55gag sequence (15-aa peptides overlapping by 11 residues) showed that DNA immunization with native gag, LAMP/gag, or DC-LAMP/gag induced responses to identical immunodominant CD4+ and CD8+ peptides. However, LAMP/gag and DC-LAMP/gag plasmids also elicited significant responses to 23 additional cryptic epitopes that were not recognized after immunization with native gag DNA. The three plasmids induced T cell responses to a total of 39 distinct peptide sequences, 13 of which were induced by all three DNA constructs. Individually, DC-LAMP/gag elicited the most diverse response, with a specific T cell response against 35 peptides. In addition, immunization with LAMP/gag and DC-LAMP/gag chimeras also promoted Ab secretion to an increased number of epitopes. These data indicate that LAMP-1 and DC-LAMP Ag chimeras follow different trafficking pathways, induce distinct modulatory immune responses, and are able to present cryptic epitopes.

Purpose: To evaluate the angulation of the maxillary sinus walls at the apical sinus region., Materials and Methods: Using preoperative computerized tomographies of 15 patients selected for sinus lift procedures, the angulation of the maxillary sinus floor was measured drawing straight lines tangential to the mesial and lateral walls. The measurements were taken from sagittal images at specific areas (i.e., second bicuspid, first molar, and second molar)., Results: The results showed that the second bicuspid sites have a sharper angulation than the second molar sites, and these second molar sites have a sharper angle than the first molar sites., Conclusion: The sharper angle observed in the second bicuspid area can influence the feasibility of schneiderian membrane evaluation when compared to the molar areas.

Heat shock proteins (HSPs) are highly conserved among different organisms. A mycobacterial HSP65 DNA vaccine was previously shown to have prophylactic and immunotherapeutic effects against Mycobacterium tuberculosis infection in mice. Here, BALB/c mice were immunized with mycobacterial DNA-hsp65 or with DNA-hsp65 and trehalose dymicolate (TDM), both carried by biodegradable microspheres (MHSP/TDM), and challenged with Leishmania (Leishmania) major. MHSP/TDM conferred protection against L. major infection, as indicated by a significant reduction of edema and parasite loads in infected tissues. Although high levels of interferon-gamma and low levels of interleukin (IL)-4 and IL-10 were detected in mice immunized with DNA-hsp65 or MHSP/TDM, only animals immunized with MHSP/TDM displayed a consistent Th1 immune response, i.e., significantly higher levels of anti-soluble Leishmania antigen (SLA) immunoglobulin G (IgG)2a and low anti-SLA IgG1 antibodies. These findings indicate that encapsulated MHSP/TDM is more immunogenic than naked hsp65 DNA, and has great potential to improve vaccine effectiveness against leishmaniasis and tuberculosis.

Durable antigen (Ag)-specific T- and B-cell anergy induced by oral tolerance is an attractive strategy for immunotherapy of allergic diseases. Here, we address the lasting effect of oral tolerance induction in naïve or primed mice to ovalbumin (OVA) on antibody production. Single feeding with OVA prior to immunization or double feeding, before and after Ag priming, in A/Sn mice, induced a long-lasting suppression of IgE, IgG1 and IgG2a responses up to 8 months after immunization. In contrast, primed-fed mice had transient IgE inhibition. Naive and double-treated mice showed marked Ag-specific unresponsiveness and scarce cytokines production. Inhibition of IL-2 and IFN-gamma secretion in naïve-fed mice were restored in the presence of anti-CD28 mAb plus Ag stimulation. The durable inhibition of Ab production in OVA-fed mice was related to the persistent decrease of B7.2 expression on B cells. Ag feeding in naive and primed status may be a prophylactic measure to avoid later Ag sensitization.

We compared the concentration of hyaluronic acid in the aqueous humor of primary open-angle glaucoma (POAG) patients and non-glaucomatous patients. Aqueous humor samples were obtained from 22 patients just before trabeculectomy for clinically uncontrolled POAG (POAG group). Aqueous humor (0.1 mL) was aspirated by inserting a 26-gauge needle into the anterior chamber. The same procedure was performed for 22 non-glaucomatous patients just before cataract surgery (control group). Immediately after collection, the aqueous humor was stored at -20 degrees C. The concentration of hyaluronic acid was determined by a sensitive, noncompetitive and nonisotopic fluoroassay. The median (range) concentrations of hyaluronic acid of the POAG and control groups were 298.4 microg L(-1) (99.0-743.7 microg L(-1)) and 545.1 microg L(-1) (145.0-2366.0 microg L(-1)), respectively. The difference in concentrations of hyaluronic acid between the groups was statistically significant (P<0.001). In conclusion, the concentration of hyaluronic acid in the aqueous humor in POAG patients is lower than in non-glaucomatous patients. Further studies are necessary to determine the role of hyaluronic acid in the pathophysiology of POAG.

The purpose of the present study was to investigate the efficacy of the decontamination protocol for bacterial removal in titanium surfaces with three different levels of roughness using a high-pressure sodium bicarbonate device for 1 minute under aseptic conditions. Group 1 was composed of 10 as-machined titanium sheets and Groups 2 and 3 of titanium sheets blasted with aluminum oxide (Al2O3, alumina) particles with different diameters: Group 2 was blasted with 65-microm particles and Group 3 with 250-microm particles. The titanium specimens were sterilized and incubated in tubes containing a suspension of Streptococcus sanguis. The colony-forming units were counted before and after the application of the decontamination protocol. The arithmetic mean roughness (R(a)) per group was: Group 1, 0.17 microm +/- 0.01; Group 2, 1.14 microm +/- 0.15; and Group 3, 3.17 microm +/- 0.23. After the contamination period, Group 1 remained with 49 x 10(3) bacterial cells, and the bacterial concentrations of Groups 2 and 3 were 11 x 10(4) and 35 x 10(5), respectively. After the application of the decontamination protocol, no viable bacteria were detected. With the increase of the surface roughness, an exponential increase in bacterial cells was observed. The results showed that the decontamination protocol treatment with a high-pressure sodium bicarbonate device efficiently removed all bacterial cells in all surfaces tested. This indicates that high-pressure sodium bicarbonate spray should be used in the maintenance phase of implant treatment.