Abscesso esplênico

Splenic abscess is a rare disease usually associated with immunosuppressed states. Its diagnosis may be difficult due to non-specific symptoms . The authors report a case of a healthy woman, 42 years old, bearer of splenic and renal abscesses , treated with antibiotics and splenectomy. A review of the literature is presented with emphasis on the diagnosis and treatment

Estudo de ß-glucanas extraídas dos fungos Geastrum saccatum e Polyporos dermoporus: características químicas e atividades biológicas

Fungal polysaccharides have received a great deal of attention due to itsbecause of their potential use in a wide rangegreat variety fromof industries. Some studies have demonstrated that polysaccharides extracted offrom basidiomycetes they have presented significant properties as anti-inflammatory, antimicrobial, antioxidant and anti-tumoral properties. In spite of thisDespite these potential properties, these mushrooms have not been insufficiently investigated, and the great number of antibiotics number produced forby these organisms suggests that they canmay be a new source of bioactives composites source. In tThe present work, reports onlated the chemical composition, potential antioxidant, antiinflammatory and citotoxycity of extracted polymers extracted offrom the fruits bodies of the fungiius Geastrum saccatum and Polyporus dermoporus, native mushrooms of the Atlantic forest inof the state of the Rio Grande do Norte, Brazil. The Cchemical analyses had revealed ademonstrated text of total sugar rates of 65% and 49%, and proteins of 7.0% for in extracts of G. saccatum and P. dermoporus extracts, respectively. The analyses ofNMR spectroscopy of RMN had demonstrated that these extracts are composites forof a complex involving β- glucans and- proteins complex. The inhibition of the formation of superoxide radicals formation was of 88.4% in G. saccatum and 83.3% in P. dermoporus, and 75 and 100% for inhibition of hydroxyls radicals inhibition. TopicalThe topic application of extracts the 10, 30 and 50 mg/kg extract in BALBc mice with cutaneous inflammation induced byfor croton oil demonstrated to inhibitedion of ear edema of ear and cells polimorfonuclears cells atin the inflamed siteplace, being this reply more effective in lower concentrations being more effective. The evaluation of the glucans of G. saccatum and P. dermoporus glucans under induced pleurisy for carrageenan-induced pleurisya of showed the antiinflammatory action of these composites., being analyzed tThe frame number in the pleural exudates and thedosage of nitric oxide dosage was also analyzed. The cytotoxic action of these polymers was analyzed throughthrough the mitochondrial function (MTT). The incubation of the glucans with mononuclear cells of the peripheral blood demonstrated that the extracted glucans extracted fromof G. saccatum havepossess a moderate cytotoxic action. These results suggest that these mushrooms possess polymers formed byfor a complex glucana-protein complex, with antiinflammatory and antioxidant actions

Avaliação do potencial de formigas (Hymenoptera: formicidae) como vetores mecânicos de bactérias do gêneroSstaphylococcus no ambiente hospitalar.

In a hospital environment, these bacteria can be spread by insects such as ants, which are characterized by high adaptability to the urban environment. Staphylococcus is a leading cause of hospital infection. In Europe, Latin America, USA and Canada, the group of coagulase negative staphylococci (CoNS) is the second leading cause of these infections, according to SENTRY (antimicrobial surveillance program- EUA). In this study, we investigated the potential of ants (Hymenoptera: Formicidae) as vehicle mechanics of Staphylococcus bacteria in a public hospital, in Natal-RN. The ants were collected, day and night, from June 2007 to may 2008, in the following sectors: hospitals, laundry, kitchen, blood bank. The ants were identified according to the identification key of Bolton, 1997. For the analysis of staphylococci, the ants were incubated in broth Tryptic Soy Broth (TSB) for 24 hours at 35 º C and then incubated on Mannitol Salt Agar. The typical colonies of staphylococci incubated for 24 hours at 35 ° C in Tryptic Soy Agar for the characterization tests (Gram stain, catalase, susceptibility to bacitracin and free coagulase). The identification of CoNS was performed through biochemical tests: susceptibility to novobiocin, growth under anaerobic conditions, presence of urease, the ornithine decarboxylation and acid production from the sugars mannose, maltose, trehalose, mannitol and xylose. The antimicrobial susceptibility examined by disk-diffusion technique. The technique of Polymerase Chain Reaction was used to confirm the presence of mecA gene and the ability to produce biofilm was verified by testing in vitro using polystyrene inert surface, in samples of resistant staphylococci. Among 440 ants, 85 (19.1%) were carrying coagulase-negative staphylococci (CoNS) of the species Staphylococcus saprophyticus (17), Staphylococcus epidermidis (15), Staphylococcus xylosus (13), Staphylococcus hominis hominis (10), Staphylococcus lugdunensis (10), Staphylococcus warneri (6), Staphylococcus cohnii urealyticum (5), Staphylococcus haemolyticus (3), Staphylococcus simulans (3), Staphylococcus cohnii cohnii (2), and Staphylococcus capitis (1). No Staphylococcus aureus was found. Among the isolates, 30.58% showed resistance to erythromycin. Two samples of CoNS (2.35%), obtained from the ant Tapinoma melanocephalum collected in the post-surgical female ward, S. Hominis hominis and S. lugdunensis harbored the mecA gene and were resistant to multiple antibiotics, and the specie S. hominis hominis even showed to be a biofilm producer. This study proves that ants act as carriers of multidrug-resistant coagulase-negative Staphylococci and biofilm producers and points to the risk of the spreading of pathogenic microorganisms by this insect in the hospital environment

Desenvolvimento de sistemas magnéticos com potencialidades terapêuticas para vetorização de fármacos

Magnetic targeting is being investigated as a means of local delivery of drugs, combining precision, minimal surgical intervention, and satisfactory concentration of the drug in the target region. In view of these advantages, it is a promising strategy for improving the pharmacological response. Magnetic particles are attracted by a magnetic field gradient, and drugs bound to them can be driven to their site of action by means of the selective application of magnetic field on the desired area. Helicobacter pylori is the commonest chronic bacterial infection. The treatment of choice has commonly been based upon a triple therapy combining two antibiotics and an anti-secretory agent. Furthermore, an extended-release profile is of utmost importance for these formulations. The aim of this work was to develop a magnetic system containing the antibiotic amoxicillin for oral magnetic drug targeting. First, magnetic particles were produced by coprecipitation of iron salts in alkaline medium. The second step was coating the particles and amoxicillin with Eudragit® S-100 by spray-drying technique. The system obtained demonstrated through the characterization studies carried out a possible oral drug delivery system, consisting in magnetite microparticles and amoxicillin, coated with a polymer acid resistant. This system can be used to deliver drugs to the stomach for treatment of infections in this organ. Another important finding in this work is that it opens new prospects to coat magnetic microparticles by the technique of spray-drying.

Nanopartíulas de magnetita com oxacilina obtida por moagem de alta energia para aplicações biomédicas

The drug targeting has been the subject of extensive studies in order to develop site-specific treatments that minimize side effects and become more effective anticancer therapy. Despite considerable interest in this class, drugs like antibiotics also have limitations, and have been neglected. Using new pharmaceutical technologies, the use of magnetic vectors appear as promising candidate for drug delivery systems in several studies. Small magnetic particles bound to the drug of interest can be modulated according to the orientation of a magnet outside the body, locating and holding in a specific site. In this work, we propose the use of High Energy Milling (HEM) for synthesis of a magnetic vector with characteristics suitable for biomedical applications by intravenous administration, and for the formation of an oxacillin-carrier complex to obtain a system for treating infections caused by Staphylococcus aureus. The results of the variation of milling time showed that the size and structural properties of the formed material change with increasing milling time, and in 60 hours we found the sample closest to the ideal conditions of the material. The vector-drug system was studied in terms of structural stability and antimicrobial activity after the milling process, which revealed the integrity of the oxacillin molecule and its bactericidal action on cultures of Staphylococcus aureus ATCC

Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi

With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of induction for IPTG was carried through, in order to verify the influence of the composition of the ways tested in the expression them recombinant proteins. On the basis of the gotten results, can be observed that the high complexity of culture medium favored the kinetic one of growth of clones recombinant (eIF, LACK), however, to if to deal with the assays submitted to the procedure of induction for IPTG, the raised complexity of culture medium did not favor the expression of recombinant proteins. On the other hand, they had been gotten resulted positive for all clones recombinant (eIF, LACK) tested, confirmed through the eletroforético profile

Avaliação de dois clones de Escherichia coli recombinante quanto ao crescimento e expressão de antígenos de Leishmania chagasi (kmp11 e P36)

Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37°C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies

Levantamento e avaliação de critérios para a ampliação de escala da produção de biossurfactantes utilizando melaço como substrato

Biosurfactants are amphiphilic molecules synthesized by microorganisms such as bacteria, yeast or filamented fungi cultivated in various carbon sources among sucrose and hydrocarbons. These molecules are composed by a hydrophilic and hydrophobic part. They operate mostly at interfaces of fluids of different polarities. Because of this characteristic, they are potentially employed in numerous industries, such as the textile, medical, cosmetics, food and mainly in the petrochemical ones. Therefore industry has interest in developing new biosurfactant production processes in high scale, in order to become them economically competitive when compared to synthetic biosurfactants. This work aims to evaluate the biosurfactant production applying a non-conventional substrate sugar cane molasses proceeding from the sugar industry thus reducing the production costs. The strain identified as AP029/GLIIA, isolated from oil wells in Rio Grande do Norte state and used in these experiments belongs to the culture collection of Antibiotics Department of UFPE. The fermentation were carried out using different conditions according to a factorial planning 24 with duplicate at center point, in which the studied factors were molasse concentration, nitrate concentration, agitation and aeration ratio. The experiments were performed in a shaker at 38ºC of temperature. Samples were withdrawn in regular periods of time of up to 72 hours of fermentation in order to analyze substrate consumption, cellular concentration, superficial tension, critical micelle dilution (CMD-1 e CMD-2) as well as extracelullar protein production. The results showed a production of 3,480 g/L of biomass, a reduction of 41% on superficial tension, 67% of substrate consumption and 0,2805 g/L of extracellular protein

Eficácia do bochecho de quitosana a 0,4% sobre o biofilme e bactérias orais

The purpose of this study was evaluate the effectiveness of the chitosan at 0.4 with high molecular weight and high deacetylation degree mouthrinse over the total decrease of the streptococci, Streptococcus mutans, lactobaci/li and over the perceptible bacterial film and gingival bleeding indices. For that, a total of 68 healthy students between 11 and 13 years old, not allergic to crustacean and not users of antibiotics or antimicrobial agent for the last three months or during the treatment, was selected. From those, thirty two individuaIs used the mouthrinse test, and thirty six, the control one. The participants rinsed 10 mL of the solutions twice a day, one during the moming (which was supervised), and another one during the aftemoon (which was not supervised), for fifteen days. The saliva collect for the microbiological analysis, as well as the perceptible bacterial film and gingival bleeding indices check, were made before the use ofthe mouthrinses (base line), immediately after the last mouthrinse on the day (zero time) and fifteen days after (fifteen time). These data were collected at school and the saliva was carried inside the ice to the laboratory. The samples were diluted, and 0.1 mL ofthe 10 -1 dilution was seeded in Rogosa SL agar, for further analysis of the total of lactobaci/lus~ 0.1 mL of the 10-4 dilution in Mitis Salivarius with bacitracin, for S. mutans analysis; and 0.1 mL of the 10-6 dilution in Mitis Salivarius for the analysis ofthe total of streptococcus. The Rogosa SL agar plates were incubated in aerobic at 37°C for 72 hours and the MSB and the MS were incubated in anaerobic in Gaspak@ jars at 37°C for 48 hours for further count ofColonies Former Units (CFUs). The assay was made in duplicate for each bacterial group analyzed. The number of CFUs transformed in LOGlO was analyzed according to the following tests: ANOV A, t of Paired and Not Paired Student, Friedman, Man-Whitney and square-qui test. On the base line, alI the variables analyzed were similar on both tested groups. On both groups, for the total of streptococcus there was no significant difference along the time and for S. mutans there was a statistic significant increase of the CFUs from the base line to the zero time. For the total of lactobaccilus there was no significant difference on the test group along the time, and on the control there was a significant increase ofthe CFUs ITom the base line to the zero time. For both groups, there was significant decrease ofthe perceptible bacterial film index along the time, and that can be explained by the mechanic effect of the mouthrinse over the bacterial film and by the participation of the students on the research which could have motivated him to a better toothbrushing (Hawthome effect). The gingival bleeding index also showed a decrease along the time, even though it was not significant. Therefore, the conclusion of this study was that the chitosan at 0.4 % mouthrinse was not effective on the CFUs reduction of the three bacterial groups analyzed, as well as on the reduction of the perceptible bacterial film and gingival bleeding indices

Adesão de células mesenquimais da medula óssea de camundongos e do ligamento periodontal de ratos a diferentes superfícies de titânio: estudo comparativo in vitro

The present experiment used cell culture to analyze the adhesion capacity of mouse mesenchymal bone marrow cells and rat periodontal ligament to different titanium surfaces. Grade II ASTM F86 titanium discs 15mm in diameter and 1.5mm thick were used and received 2 distinct surface treatments (polished and cathodic cage plasma nitriding). The cells were isolated from the mouse bone marrow and rat periodontal ligament and cultured in α-MEM basic culture medium containing antibiotics and supplemented with 10% FBS and 5% CO2, for 72 hours at 37ºC in a humidified atmosphere. Subculture cells were cultured in a 24-well plate with a density of 1 x 104 cells per well. The titanium discs were distributed in accordance with the groups, including positive controls without titanium discs. After a 24-hour culture, the cells were counted in a Neubauer chamber. The results show that both the mouse mesenchymal bone marrow cells and rat periodontal ligament cells had better adhesion to the control surface. The number of bone marrow cells adhered to the polished Ti surface was not statistically significant when compared to the same type of cell adhered to the Ti surface treated by cathodic cage plasma nitriding. However a significant difference was found between the control and polished Ti groups. In relation to periodontal ligament cell adhesion, a significant difference was only found between the control and plasma-treated Ti surfaces. When comparing equal surfaces with different cells, no statistically significant difference was observed. We can therefore conclude that titanium is a good material for mesenchymal cell adhesion and that different material surface treatments can influence this process

A influência da criopreservação nas células mesenquimais indiferenciadas do ligamento periodontal de humanos análise comparativa in vitro

Cryopreservation is a process where cells or biological tissues are preserved by freezing at very low temperatures and aims to cease reversibly, in a controlled manner, all the biological functions of living tissues, i.e., maintain cell preservation so that it can recover with high degree of viability and functional integrity. This study aimed to evaluate the influence of cryopreservation on the mesenchymal stem cells originating from the periodontal ligament of human third molars by in vitro experiments. Six healthy teeth were removed and the periodontal cells grown in culture medium containing α-MEM supplemented with antibiotics and 15% FBS in a humidified atmosphere with 5% CO2 at 37° C. Cells isolated from each sample were divided into two groups: Group I - immediate cell culture (not fresh cryopreserved cells) and Group II - cell cryopreservation, during a period of 30 days. Analyses of rates of cell adhesion and proliferation in different groups were performed by counting the cells adhered to the wells, in intervals of 24, 48 and 72 hours after the start of cultivation. The number of cells in each well was obtained by counting viable cells with the use of hemocytometer and the method of exclusion of cells stained by trypan blue. The difference between groups for each of the times was analyzed by Wilcoxon test. Regarding the temporal evolution for each group, analysis was done by Friedman's test to verify the existence of differences between times and, when it existed, the Wilcoxon penalty was applied. The results showed no statistically significant difference between the two groups analyzed in this study. Therefore, we conclude that the cryopreservation process, after a period of 30 days, did not influence the cell type studied, and there was no difference in growth capacity in vitro between the groups

Prevalência e susceptibilidade antimicrobiana de Staphylococcusspp. em quadros de saúde e doença periodontal

The aim of this study was determine the prevalence and antimicrobial susceptibility of Staphylococcus spp. from patients with periodontal disease and periodontally healthy, correlate them with factors to host, local environment and traits of the diseases. To this, thirty adults from 19 to 55 years old were selected. They had not periodontal treatment and no antibiotic or antimicrobial was administered during three previous months. From these individuals, sites periodontally healthy, with chronic gingivitis and/or periodontitis were analyzed. Eighteen subgingival dental biofilm samples were collected through sterile paper points being six from each tooth randomly selected, representing conditions mentioned. They were transported to Oral Microbiology laboratory, plated onto Mannitol Salt Agar (MSA) and incubated at 370C in air for 48 h. Staphylococcus spp. were identified by colonial morphology, Gram stain, catalase reaction, susceptibility to bacitracin and coagulase activity. After identification, strains were submitted to the antibiotic susceptibility test with 12 antimicrobials, based on Kirby-Bauer technique. To establish the relation between coagulase-negative Staphylococcus (CSN) presence and their infection levels and host factors, local environment and traits of diseases were used Chi-square, Mann-Whitney and Kruskal-Wallis tests to a confidence level of 95%. 86,7% subjects harbored CSN in 11,7% periodontal sites. These prevalence were 12,1% in healthy sites, 11,7% in chronic gingivitis, 13,5% in slight chronic periodontitis, 6,75% in moderate chronic periodontitis and in sites with advance chronic periodontitis was not isolated CSN, without difference among them (p = 0,672). There was no significant difference to presence and infection levels of CSN as related to host factors, local environm ent and traits of the diseases. Amongst the 74 samples of CSN isolated, the biggest resistance was observed to penicillin (55,4%), erythromycin (32,4%), tetracycline (12,16%) and clindamycin (9,4%). 5,3% of the isolates were resistant to oxacilin and methicillin. No resistance was observed to ciprofloxacin, rifampicin and vancomycin. It was concluded that staphylococci are found in low numbers in healthy or sick periodontal sites in a similar ratio. However, a trend was observed to a reduction in staphylococci occurrence toward more advanced stages of the disease. This low prevalence was not related to any variables analyzed. Susceptibility profile to antibiotics demonstrates a raised resistance to penicillin and a low one to methicillin. To erythromycin, tetracycline and clindamycin was observed a significant resistance

Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação

Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies