Polyphenols from guaraná after in vitro digestion: Evaluation of bioacessibility and inhibition of activity of carbohydrate-hydrolyzing enzymes.

Guaraná (Paullinia cupana) is a typical product from Amazon biota that exerts antioxidant capacity due to the presence of phenolic compounds, such as catechin, epicatechin and proanthocyanidins. The objective of this study was to evaluate the potential inhibitory activity of guaraná extracts, after digestion in vitro, on carbohydrates-metabolism enzymes and to assess the bioacessibility of guaraná polyphenols. The guaraná samples before and after enzymatic digestion, were compared for total phenolic content and phenolic profile. Furthermore, we investigated the uptake of polyphenols from guarana, using Caco-2 cells, and the effect of digested guaraná on carbohydrate metabolism enzymes. The amount of total phenolic compounds extracted from guaraná decreased after digestion in vitro, and native phenolics were not identified after cell permeation. On the other hand, polyphenols from guaraná were able to inhibit α-glucosidase and α-amylase activities. In conclusion, guaraná can be considered as a dietary source with anti-hyperglycemic potential.

Matrix metalloproteinase-8 levels in periodontal disease patients: A systematic review.

Periodontal disease is characterized as a disorder of the oral microbiota resulting in an immune response which, in turn, leads to the destruction of periodontal tissue. Matrix metalloproteinase-8 (MMP-8) has been reported as the major metalloproteinase involved in periodontal disease, being present at high levels in gingival crevicular fluid and salivary fluid (SF). The aim of this systematic review was to evaluate the scientific literature regarding the expression of MMP-8 in gingival crevicular fluid and SF in patients with periodontal disease, analyzing its validity as a possible biomarker in the diagnosis of periodontal disease. A systematic review of the literature was performed using the PubMed/Medline, CENTRAL and Science Direct databases. Studies concerning the use of MMP-8 in the diagnosis of periodontal disease that evaluated its effectiveness as a biomarker for periodontal disease were selected. The search strategy provided a total of 6483 studies. After selection, six articles met all the inclusion criteria and were included in the present systematic review. The studies demonstrated significantly higher concentrations of MMP-8 in patients with periodontal disease compared with controls, as well as in patients presenting more advanced stages of periodontal disease. The findings on higher MMP-8 concentrations in patients with periodontal disease compared with controls imply the potential adjunctive use of MMP-8 in the diagnosis of periodontal disease.

Participation of osteoclastogenic factors in immunopathogenesis of human chronic periapical lesions.

BACKGROUND: Chronic periapical lesions (CPLs) are common lesions of the oral cavity and are the result of caries, tooth fracture, iatrogenic causes, or factors causing contamination and pulp necrosis. Inflammatory cells participate in the expansion of CPLs by releasing factors that stimulate or inhibit osteolytic activity. The objective of this study was to investigate the participation of RANKL, TNF-α, cathepsin K, IL-33, and OPG in the development of radicular cysts (RCs) and periapical granulomas (PGs). METHODS: Paraffin-embedded sections of 30 RCs and 22 PGs were submitted to immunohistochemistry. RESULTS: Immunoexpression of the proteins studied was observed in the epithelium and capsule of RCs, as well as in connective tissue of PGs. The expression of the osteoclastogenic factors studied differed significantly in RCs and PGs (P < .001), with lower expression of OPG in RCs. In PGs, the lowest expression was observed for cathepsin K. Comparison of the 2 lesions showed a similar participation of RANKL and IL33, while a significant difference was observed for OPG (P < .001), TNF-α (P = .002), and cathepsin K (P = .016). No association of the expression of the proteins with lesions size was observed. CONCLUSIONS: This study demonstrated the participation of RANKL, TNF-α, IL-33, cathepsin K, and OPG in the development of RCs and PGs, with emphasis on the highest immunoreactivity of cathepsin in RCs and TNF-α and OPG in PGs. OPG possibly determines the slower growth of PGs compared to RCs.

Brain Death: Knowledge of Future Brazilian Physicians.

BACKGROUND: Thousands of people die on the waiting list for transplants. The shortage of organs and tissues for transplantation is considered a serious global problem. Brain death (BD) is the main source of organs for transplantations. OBJECTIVE: Given that BD is assessed by medical staff and that this diagnosis is ethically relevant, the goal of this work was to analyze the knowledge of 5th- and 6th-year medical students of Southern Brazil regarding the process of determining BD. METHOD: This observational, cross-sectional study was conducted between September 2013 and March 2014. RESULTS: A total of 635 students participated in the study, with 337 (53.1%) from public and 298 (46.9%) from private schools. Approximately 333 (52.4%) attended the 6th year of the medical program. The mean age of the students was 25.4 ± 3.56 years, with a prevalence of females of 54.6% (347) and a prevalence of Catholic religion of 61.7% (392); 84.2% of the students reported not feeling confident performing the BD protocol. A low percentage (15.7%) correctly answered 75% or more of the questions about BD, a criterion used to determine sufficient knowledge of BD. Approximately 45% (282) of respondents were not able to identify the patients who were candidates for the BD protocol. Analyzing the medical students' knowledge at the end of the program can provide information about the quality of undergraduate education on the topic of determining BD. CONCLUSION: It was concluded that 5th- and 6th-year medical students of Paraná State have little knowledge about the BD protocol.

Early Vascular Thrombosis After Kidney Transplantation: Can We Predict Patients at Risk?

BACKGROUND: Renal transplant is the therapy of choice for patients with chronic renal disease. In recent years, improvement in immunosuppressive drugs reduced early graft loss associated with acute rejection. However, vascular thrombosis, accounting for 5% of early graft loss, can sensitize the recipient for human leukocyte antibodies, reducing the chance for a second transplant. The aim of this study was to identify risk factors for vascular thrombosis in a single transplant center, to design specific prevention protocol. METHODS: This was a retrospective, case-control study. From the Renal Transplant Unit database, we identified 21 cases of vascular thrombosis in recipients of kidneys from deceased donors. Recipients from the contralateral kidney from the same donor, without vascular complications, were assigned to the control group. Data analyzed included donor, recipient, transplant surgery, and post-operative follow-up. The local ethics committee approved the protocol. RESULTS: Thrombosis and control groups were comparable for recipient characteristics, cold ischemia time, organ side (right or left), and site of arterial anastomosis. We observed an increased risk for vascular thrombosis in kidneys with multiple veins (odds ratio, 11.32; P = .03). Organ retrieval surgery complications, such as vascular lesions or heterogeneous perfusion, despite normal pre-implantation biopsy, were considered risk factors for vascular thrombosis within the first post-operative day (odds ratio, 7.1; P = .03). CONCLUSIONS: In this series, multiple renal vein and organ retrieval surgery complications were risk factors for early vascular thrombosis.

Analysis of tryptase-positive mast cells and immunoexpression of MMP-9 and MMP-13 in periapical lesions.

AIM: To evaluate the immunoexpression of tryptase, MMP-9 and MMP-13 in periapical lesions, correlating them with the type of lesion, intensity of the inflammatory infiltrate and thickness of the epithelial lining. METHODOLOGY: Twenty periapical granulomas (PGs), twenty radicular cysts (RCs) and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using antitryptase, anti-MMP-9 and anti-MMP-13 antibodies. Immunoexpression of MMP-9 and MMP-13 was quantitatively evaluated both in the connective tissue of all lesions and in the epithelial lining of RCs and RRCs. Tryptase-positive mast cells were counted only in the connective tissue. The data were analysed using Kruskal-Wallis and Fisher's exact tests, as well as Spearman's correlation test (P < 0.05). RESULTS: In comparison with RCs and RRCs, PGs exhibited higher immunoexpression of tryptase, MMP-9 and MMP-13 (P = 0.002, P = < 0.001 and P < 0.001, respectively). In comparison with lesions with inflammatory infiltrates grades I and II, lesions with inflammatory infiltrate grade III had higher median percentages of MMP-13-positive cells (P = 0.003) and a tendency for higher expression of MMP-9 (P = 0.059). No significant difference was observed between the expression of the studied markers and epithelial thickness (P > 0.05). There were positive correlations between the number of tryptase-positive mast cells and the immunoexpression of MMP-9, as well as between the immunoexpression of MMP-9 and MMP-13. CONCLUSION: A larger number of tryptase-positive mast cells and greater enzymatic activity of MMP-9 and MMP-13 were found in PGs compared to RCs and RRCs. These findings are a characteristic of the dynamics of periapical diseases.

Immunoexpression of tumour necrosis factor-α, interleukin-1α and interleukin-10 on odontogenic cysts and tumours.

AIM: To analyse the immunoreactivity of IL-1α, TNF-α and IL-10 in odontogenic cysts and tumours and to investigate possible associations with established biological behaviours of these different lesions. METHODOLOGY: Immunohistochemical expression of anti-IL-1α, anti-TNF-α and anti-IL-10 antibodies was assessed on epithelium and mesenchyme of 20 radicular cysts (RCs), 20 residual cysts (RECs), 20 dentigerous cysts (DCs), 18 solid ameloblastomas (SAs), 20 keratocystic odontogenic tumours (KCOTs) and 15 dental follicles (DFs). Comparative analysis of data was performed using the nonparametric Wilcoxon signed-rank test and Kruskal-Wallis's test. RESULTS: Significantly greater expression of IL-1α in the epithelium was noted in RC, KCOT and SA (P = 0.01), whilst IL-10 and TNF-α was in the epithelium of RC, DC and KCOT (P < 0.01). In the mesenchyme, significantly greater immunopositivity was observed for IL-1α, IL-10 and TNF-α in KCOT, DC and RC (P < 0.01). In epithelial and mesenchymal tissues, there were a significant number of cases of RC and DC with IL-1α < IL-10 ratio (P < 0.01), whilst SA and KCOT showed IL-1α > IL-10 (P < 0.01). There was a significantly greater percentage of DF, DC and KCOT with TNF-α > IL10 ratio (P < 0.01). CONCLUSION: These results suggest involvement of the proteins in the pathogenesis of odontogenic cysts and tumours, with emphasis on the highest immunoreactivity of osteolysis stimulating factors in tumours with aggressive biological behaviour, such as SA and KCOT.

Tuning Fe3O4 nanoparticle dispersion through pH in PVA/guar gum/electrospun membranes.

Polyvinyl Alcohol (PVA)/guar gum (GG) membranes with different loads of paramagnetic iron oxide Fe3O4 nanoparticles were successfully electrospun using both non-alkaline and alkaline stock solutions. The nanoparticle homogeneity distribution was clearly enhanced in fibers obtained from alkaline stock solutions. This is mainly due to the interaction between GG and the metallic ion, which also leads to further dispersion of remained uncoated nanoparticles in the mixture. It was also noticed that GG favors nanoparticle stability in the mixture and contributes to nanoparticle encapsulation. X-ray results showed that all membranes were semi-crystalline. FTIR-ATR spectra showed that Fe-O absorption band intensity improved with increasing nanoparticle load, reaching saturation at 3.5mg/ml Fe3O4 concentration under alkaline conditions. VSM analyses showed that the nanoparticles are paramagnetic and were successfully incorporated by the fibers. In vitro biocompatibility tests using L929 cells indicates adequate levels of cytotoxicity and cell adhesion/proliferation assays for both membranes obtained from non-alkaline and alkaline stock solutions. Therefore, they have potential for biomedical applications as biodegradable wound dressing.

Organ and tissue donation for transplantation from fatal trauma victims.

INTRODUCTION: The aim of this study was to analyze multiple aspects of organ donation after mortality owing to trauma-related causes. METHODS: We completed a cross-sectional, retrospective, descriptive study that used data were extracted from monthly death reports and clinical records of fatal external injury victims 2-70 years of age who were assisted at 1 of 4 hospitals in the municipality of Maringá and the surrounding metropolitan area in Paraná state, Brazil, in 2012. RESULTS: A total of 871 individuals within the age range for donation died, 15.4% owing to external causes of injury. Of these 134 cases, 80.6% were male and 52.23% were 18-40 years old. A total of 57% of deaths were owing to traffic accidents, 58% of which involved motorcycles. Aggression was a factor in 25% of deaths, with 55% owing to firearm wounds. Approximately 52% of families consented to organ donation when asked. A total of 58% of families who refused consent did not indicate a specific reason. At 62%, the majority of refusals involved brain-dead patients. Approximately 58% of the families did not receive a request for consent. Out of these 78 cases, 83% showed contraindications, 44% had sepsis, 40% hemodilution, and 10% logistical/infrastructural problems. CONCLUSIONS: The highest proportion of non donor cases was owing to the lack of a formal request for consent from the family. Thus, continued training for health care professionals on the donation-transplantation process and early identification of potential donors and appropriate organ maintenance is necessary.

Analysis of the frequency and nature of hyaline ring granulomas in inflammatory odontogenic cysts.

AIM: To determine the prevalence of hyaline ring granulomas (HRGs) in a large case series of inflammatory odontogenic cysts, and to investigate the nature of these structures. METHODOLOGY: All records from the patients diagnosed with inflammatory odontogenic cysts between January 1970 and April 2009 were reviewed. Histologic sections were evaluated by light microscopy and cases with HRGs for which sufficient biological material was available were submitted to histochemical analysis (Masson's trichrome) and immunohistochemistry (CD34, CD68 and collagen IV). RESULTS: Twenty-two (3.3%) of the 661 cases of inflammatory odontogenic cysts diagnosed during the study period presented HRGs. The relative frequency of HRGs was higher amongst residual radicular cysts (6.1%), followed by paradental cysts (5.6%) and radicular cysts (3.0%). HRGs appeared as roughly circular homogeneous/fibrillar masses in 14 (63.6%) cases and as round structures enclosing amorphous material in 3 (13.6%) cases. Most (77.8%) roughly circular homogeneous/fibrillar masses were positive for collagen, whereas all (100.0%) round structures enclosing amorphous material were negative for this protein. Immunohistochemistry showed that most mononucleated cells and all multinucleated giant cells were positive for CD68, but negative for CD34, in all cases. In addition, collagen IV immunostaining was negative in amorphous structures and weakly positive in homogeneous/fibrillar masses. CONCLUSIONS: The present results suggest a very low frequency of HRGs in inflammatory odontogenic cysts and support the hypothesis that these structures arise from the implantation of foreign material, most likely food particles of plant or vegetable origin. The diverse microscopic features of HRG possibly represent different developmental stages of this structure.

Causes of organ donation failure in Brazil.

INTRODUCTION: There has been a great improvement in transplantation medicine in Brazil in the last 2 decades. However, there remain several barriers regarding notification of brain and cardiac death as well as completion of the donation process. METHODS: This retrospective study was performed between January 2008 and December 2010. We reviewed all deaths in a University Hospital, observing the causes of non-notification to the State Transplantation Authority and non-donations. RESULTS: There were 41 notifications of brain death resulting in donation in only 19.5% of those cases. Cardiac death was diagnosed in 21 patients, resulting in 52.4% donations. The main cause for non-donation were family refusal (37.2%), infectious diseases (30.2%), and clinical contraindications (32.6%). Most of the missed possible donors occurred during the night (54.8%) and in the emergency room (80.9%). CONCLUSION: There is an urgent need for better education of the Brazilian population about organ donation and brain death definitions. Other identified problems include lack of uniformity in brain death determinations among hospitals, rigid contraindications to donation in the State of Parana, physician unawareness or disbelief about brain death diagnostic criteria, and lack of structure of our Hospital.

Immunoexpression of MMPs-1, -2, and -9 in ameloblastoma and odontogenic adenomatoid tumor.

OBJECTIVE: The aim of this study was to evaluate and compare the expression of metalloproteinases-1, -2, and -9 in solid ameloblastoma and adenomatoid odontogenic tumor. METHODS: A total of 20 cases of solid ameloblastoma and 10 cases of adenomatoid odontogenic tumors were selected and immunohistochemically assessed. Metalloproteinases-1, -2, and -9 immunoexpression and their distribution pattern were noted and semiquantitatively scored. The scores obtained were statistically analyzed. RESULTS: Matrix metalloproteinase (MMP)-1 showed a predominant expression in both tumors and was found in stroma and parenchyma. For MMP-2, there was a varied expression, with 80% and 60% of immunoreactive tumor cells in ameloblastoma and adenomatoid odontogenic tumor respectively. Regarding stromal cells, 65% of ameloblastomas and 80% of adenomatoid odontogenic tumors showed positivity. There was immunoexpression of the MMP-9 in parenchymal and stromal cells in all cases of both tumors analyzed. A statistically significant difference in the expression of MMP-1 in relation to the expression of MMP-2 and -9 in ameloblastomas (P < 0.001) was observed. CONCLUSION: The results suggest that these metalloproteinases are related to growth and progression of tumors analyzed, and particularly in ameloblastoma, its highest aggressiveness may be, in part, a result of the active participation of the stromal cells and their products, such as the MMPs studied.

Excessive Pap smears due to opportunistic cervical cancer screening.

The study aimed to analyze the Pap smears carried out for cervical cancer screening according to Ministry of Health guidelines. All smear tests carried out within the public health system in Campinas in 2003 were analyzed. All tests that did not conform to the guidelines were considered excessive. The guidelines recommend screening once every three years for all women aged 25 to 59 after they have received two negative smears. This study showed that the majority of women initiated screening prior to 25 years of age and the periodicity was predominantly annual, followed by biannual tests. In conclusion, 63.4% of tests were excessive. The screening coverage was 14.76%, but if all the tests had been performed as recommended, the final coverage over three years could have reached 65.4%. Thus it is possible to increase the coverage with the available resources since the screening works like an organized program.

Phylogeny of snake trypanosomes inferred by SSU rDNA sequences, their possible transmission by phlebotomines, and taxonomic appraisal by molecular, cross-infection and morphological analysis.

Blood examination by microhaematocrit and haemoculture of 459 snakes belonging to 37 species revealed 2.4% trypanosome prevalence in species of Viperidae (Crotalus durissus and Bothrops jararaca) and Colubridae (Pseudoboa nigra). Trypanosome cultures from C. durissus and P. nigra were behaviourally and morphologically indistinguishable. In addition, the growth and morphological features of a trypanosome from the sand fly Viannamyia tuberculata were similar to those of snake isolates. Cross-infection experiments revealed a lack of host restriction, as snakes of 3 species were infected with the trypanosome from C. durissus. Phylogeny based on ribosomal sequences revealed that snake trypanosomes clustered together with the sand fly trypanosome, forming a new phylogenetic lineage within Trypanosoma closest to a clade of lizard trypanosomes transmitted by sand flies. The clade of trypanosomes from snakes and lizards suggests an association between the evolutionary histories of these trypanosomes and their squamate hosts. Moreover, data strongly indicated that these trypanosomes are transmitted by sand flies. The flaws of the current taxonomy of snake trypanosomes are discussed, and the need for molecular parameters to be adopted is emphasized. To our knowledge, this is the first molecular phylogenetic study of snake trypanosomes.

Immunohistochemical assessment of extracellular matrix components in syndrome and non-syndrome odontogenic keratocysts.

OBJECTIVE: Investigate the immunohistochemical distribution of fibronectin, tenascin, laminin and collagen IV in syndrome (SOKC) and non-syndrome odontogenic keratocysts (NSOKC). MATERIALS AND METHODS: Ten cases of SOKC and five of NSOKC were selected and streptoavidin-biotin technique was applied. The specimens were analyzed taking into account the following evaluation parameters: presence, continuity and thickness in the basement membrane and intensity, distribution and association with inflammatory cells in the cyst wall. RESULTS: Differences could be detected regarding tenascin, fibronectin and collagen IV between the SOKC and NSOKC. Tenascin was present in all cases along the basement membrane in SOKC and in five cases of NSOKC predominated negative areas. Furthermore, tenascin distribution was focal in the cyst wall in SOKC whereas in NSOKC it was diffuse. Concerning fibronectin, it was detected as a discontinuous band when present in SOCK and as a continuous band in NSOKC. Collagen IV was not present in the majority of the cases in SOKC. Negative areas for laminin predominated in the basement membrane in both groups. CONCLUSIONS: These findings show differences between the immunohistochemical expression of tenascin, fibronectin and collagen IV which might indicate a more aggressive biological behavior of SOKC as compared with NSOKC.

The future of nanofabrication and molecular scale devices in nanomedicine.

Nanotechnology is engineering and manufacturing at the molecular scale, and the application of nanotechnology to medicine is called nanomedicine. Nanomedicine subsumes three mutually overlapping and progressively more powerful molecular technologies. First, nanoscale-structured materials and devices that can be fabricated today hold great promise for advanced diagnostics and biosensors, targeted drug delivery and smart drugs, and immunoisolation therapies. Second, biotechnology offers the benefits of molecular medicine via genomics, proteomics, and artificial engineered microbes. Third, in the longer term, molecular machine systems and medical nanorobots will allow instant pathogen diagnosis and extermination, chromosome replacement and individual cell surgery in vivo, and the efficient augmentation and improvement of natural physiological function. Current research is exploring the fabrication of designed nanostructures, nanoactuators and nanomotors, microscopic energy sources, and nanocomputers at the molecular scale, along with the means to assemble them into larger systems, economically and in great numbers.

Nanodentistry.

BACKGROUND: Nanodentistry will make possible the maintenance of comprehensive oral health by involving the use of nanomaterials, biotechnology (including tissue engineering) and, ultimately, dental nanorobotics (nanomedicine). RESULTS: When the first micrometer-sized dental nanorobots can be constructed within 10 to 20 years, these devices will allow precisely controlled oral analgesia, dentition replacement therapy using biologically autologous whole replacement teeth manufactured during a single office visit, and rapid nanometer-scale precision restorative dentistry. CLINICAL IMPLICATIONS: New treatment opportunities may include dentition renaturalization, permanent hypersensitivity cure, complete orthodontic realignments during a single office visit, covalently bonded diamondized enamel and continuous oral health maintenance through the use of mechanical dentifrobots.

Distribution of phlebotomine sand flies (Diptera: Psychodidae) in the state of Rio Grande do Norte, Brazil.

Visceral and cutaneous leishmaniasis are major endemic diseases in northeast Brazil. The objective of the current study was to determine the species and geographic distribution of potential sand fly vectors of Leishmania in the state of Rio Grande do Norte. Sand flies were captured using CDC light traps in 30 municipalities distributed throughout the 8 geographic zones of the state. Twelve Lutzomyia species were identified. Lutzomyia longipalpis Lutz & Neiva was the most prevalent and accounted for 85.59% of the sand fly captured. The remaining species were distributed as follows: L. evandroi Costa Lima & Antunes (10.83%), L. oswaldoi Mangabeira (0.99%), L. sallesi Galvão & Coutinho (0.58%), L. intermedia Lutz & Neiva (0.53%), L. lenti Mangabeira (0.53%), L. migonei França (0.49%), L. walkeri Newstead (0.24%), L. goiana Martins, Falcão & Silva (0.15%), L. samueli Deane (0.04%), and L. capixaba Dias, Falcão, Silva & Martins (0.03%), and L. peresi Mangabeira (0.01%). L. longipalpis, which is known to be a vector of Leishmania chagasi Cunha & Chagas (L. donovani chagasi), was captured in 93% of municipalities distributed across all geographical areas of the state and its distribution was independent of obvious climatic and topographic parameters. It was identified in all municipalities where human visceral leishmaniasis had been reported. In contrast, climate and topography appeared to be important for other Lutzomyia species. For example, L. intermedia and L. migonei, which are known to transmit Leishmania braziliensis Viana, were geographically restricted. They were captured in municipalities where cases of cutaneous and mucosal leishmaniasis had been reported. The widespread distribution of L. longipalpis, its adaptation to peridomicillary settings, and its ability to transmit L. (d.) chagasi suggest that a large number of persons may be at risk of acquiring visceral leishmaniasis in the state of Rio Grande do Norte, Brazil.