Periapical tissue healing after post space preparation with or without use of a protection plug and root canal exposure to the oral environment. Study in dogs

The purpose of this study was to evaluate the influence of coronal leakage on the healing of dogs' periapical tissues after root canal filling, post space preparation and protection or not with a temporary sealer plug. Forty root canals of dogs' teeth were instrumented and filled by the lateral condensation technique with gutta-percha points and Endomethasone or CRCS sealers. After post space preparation, the remaining filling material was protected or not with a plug of temporary Coltosol sealer and exposed to the oral environment for 90 days. Thereafter, the animals were sacrificed and the specimens were removed and prepared for histomorphological and histobacteriological analysis. The findings revealed 35% of microbial leakage in the groups without plugs and 15% of leakage in the groups with plugs. Statistical analysis showed that the use of a Coltosol plug improved significantly the histomorphological results regardless of the type of root canal sealer (p=0.05) and that CRCS and Endomethasone sealers showed similar results (p>0.05).

Multidrug-resistant urinary tract isolates of Escherichia coli from Ribeirão Preto, São Paulo, Brazil

Multiple resistances to antimicrobial drugs arising in Escherichia coli isolates may complicate therapeutic management of urinary tract infection (UTI) by this organism. In order to assess the multidrug resistance (MDR) among urinary E. coli isolates, we have tested 11 antimicrobial drugs against 67 isolates from outpatients attended in a tertiary-care teaching hospital and of 78 isolates from a municipal health unit, respectively in Ribeirão Preto, State of São Paulo, Brazil. Seventy-six percent and 22% of the isolates from the tertiary-care hospital and the municipal unit, respectively, were resistant to three or more different classes of agents, and were considered to present MDR. Among the isolates from the hospital patients, 73.0%, 65.0%, 58.0%, 58.0% and 31.0% were resistant to tetracycline, ampicillin, cephalothin, trimethoprim-sulfamethoxazole (TMP/SMX) and norfloxacin, respectively; resistance from the municipal unit patients were 31.0%, 37.0%, 8.0%, 29.0% and 12.0% respectively, to the same drugs. The predominant phenotype among the MDR isolates presented is ampicillin, TMP/SMX and tetracycline resistance. The high prevalence of drug resistance among UTI patients calls for continuous surveillance to assure effective control of this infection. © 2007 by The Brazilian Journal of Infectious Diseases and Contexto Publishing. All rights reserved.

Tuberculose e o estudo molecular da sua epidemiologia

Systems that can distinguish epidemiologically-related Mycobacterium tuberculosis strains from unrelated ones are extremely valuable. Molecular biology techniques have allowed a great deal of information to be acquired about the infectious disease tuberculosis (TB) that was very hard or impossible to obtain by conventional epidemiology. A typing method based on bacterial DNA genome differences, known as RFLP (restriction fragment length polymorphism), is widely used to discriminate strains in the epidemiologic study of TB. However, RFLP is laborious and there is a tendency to replace it by other methods. Thus, other DNA sequences have been employed as epidemiological markers, as in Spoligotyping, a fast technique based on PCR followed by differential hybridization of amplified products. The polymorphism observed among different isolates is probably the product of strain-dependent recombination. MIRU (mycobacterial interspersed repetitive unit) typing is a reproducible and fast assay, involving the generation of genotypes based on the study of 12 loci containing VNTRs (variable-number tandem repeats) in strains of the M. tuberculosis complex. It compares strains from different geographic areas and allows the movement of individual lineages to be tracked, as in RFLP. This approach enables a greater number of isolates to be analyzed, leading to the identification of a larger number of foci of transmission within the population and thus to improved ways of slowing the progress of the disease.

Espectro de agentes etiológicos associados a surtos de doenças transmitidas por alimentos em núcleos receptores turísticos de três regiões geográficas do Estado de São Paulo

Foodborne and waterborne diseases are spread by the consumption of food or water contaminated with bacteria and/or their toxins, viruses, parasites or chemicals. The aim of the research reported here was to establish the spectrum of etiologic agents of foodborne outbreaks at 15 tourist resorts in three geographic regions of the State of Sao Paulo (Brazil). The study was based on the cases reported to the Epidemic Surveillance Center (CVE) of the Public Health Authority of the State of São Paulo (SES), from 2002 to 2005. The tourist centers were chosen at random in three regions of the state (Capital, Interior and Coast) and offered the following attractions: events, agribusiness, cultural history, shopping, town center, gastronomy, health and leisure, sun and sea. Among the bacteria, the results showed that Salmonella spp. were most frequently associated with outbreaks, followed by Shigella spp., enterotoxigenic Escherichia coli, Staphylococcus aureus, Clostridium perfringens, Bacillus cereus and Campylobacter spp. Viruses (Rotavirus and Hepatitis A) played a part in many of the cases, while the frequency of parasites and worm infestations was low in the foodborne disease outbreaks at these resorts. The mixed foods (rice, beans, liver, potatoes, barbecue, juice), fish and poultry were the three commonest vehicles implicated in the outbreaks.

Plesiomonas shigelloides: Um enteropatógeno emergente?

Plesiomonas shigelloides is a Gram-negative rod-shaped bacterium, of the family Enterobacteriaceae, which has been isolated from freshwater and salt water, freshwater fish, shellfish and many species of animals. Most human P. shigelloides infections are suspected to be waterborne. The organism can be found in untreated water used as drinking water, in recreational water, or in water used to rinse food that is consumed without cooking or heating. The ingestion of P. shigelloides does not always cause illness in the host animal, and the organism may be present temporarily as a transient, noninfectious member of the intestinal flora. It has been isolated from the stools of patients with diarrhea,but it is also sometimes isolated from healthy individuals. P. shigelloides has been implicated in gastroenteritis, usually a self-limiting disease characterized by fever, chills, abdominal pain, nausea, diarrhea or vomiting; in severe cases the diarrhea may be yellowish-green, foamy and tinged with blood. The bacteria may also cause extra-intestinal infection. Furthermore, it can produce toxins and may be invasive. The evidence in favor of considering P. shigelloides as an enteropathogen is not totally convincing. Although it has been isolated from patients with diarrhea and incriminated in some outbreaks involving contaminated water and food, it was not possible, in many P. shigelloides samples associated with gastrointestinal infections, to identify a definite mechanism of virulence.

Produção deficiente de citocinas Th1 em camundongos BALB/c jovens

O objetivo do presente estudo foi comparar a produção de IFN-γ, IL-12 e IL-4 entre camundongos jovens (5, 12 e 19 dias de idade) e adultos (30 dias de idade). As avaliações foram feitas por estimulação, in vitro, de células esplênicas com Concanavalina A (ConA) , Staphylococcus aureus (S. aureus) e lipopolissacarídeo (LPS). Diferentes concentrações de cada estímulo foram testadas e os sobrenadantes das culturas foram coletados após 48 horas de incubação e as concentrações de IFN-γ, IL-12 e IL-4 determinadas por ELISA. Células de camundongos jovens e adultos produziram níveis igualmente elevados de IFN-γ após estímulo com ConA. Somente animais adultos produziram IFN-γ em resposta ao estímulo com S. aureus. Em culturas estimuladas com LPS, a produção desta citocina foi baixa e similar nos animais jovens e significativamente elevada nos animais adultos. Somente células de animais adultos estimuladas com S. aureus foram capazes de produzir IL-12. O único estímulo capaz de induzir níveis detectáveis de IL-4 foi ConA, sendo que estes níveis foram mais elevados nos animais com 12 e 19 dias de idade em comparação com animais neonatos e adultos. A diminuição das doses ótimas dos estímulos não mudou o perfil de produção de cada citocina nos animais jovens. Estes resultados permitem concluir que a idade afeta a produção de citocinas: ocorre maior produção de IL-4 em camundongos jovens e maior produção de IL-12 e IFN-γ em animais adultos. Estas informações são importantes devido ao papel destas citocinas na polarização das respostas imunes nos sentidos Th1 e Th2. Palavras-chave: camundongo; citocina; interferon-gama; interleucina-4; interleucina-12.

Detecção da produção de slime por estafilococos coagulase-negativa isolados de cateter venoso central

Slime production is an important virulence factor of coagulase-negative Staphylococcus spp., allowing them to attach to smooth surfaces of biomaterials, and it has been associated with infections of implanted medical devices. In the present study the production of slime capsules in 27 strains of coagulase-negative Staphylococcus was investigated by culture in Congo Red agar (77.7% positivity), spectrophotometric or microplate method (81.4% positivity) and scanning electron microscopy (88.9% positivity). The resistance of coagulase-negative strains of Staphylococcus to various antimicrobial agents was also determined by agar disk diffusion. The proportion of strains resistant to penicillin G, oxacillin, erythromycin, clindamycin and gentamicin among the slime-producing staphylococci was 88.9%, 70.4%, 81.5%, 66.7% and 59.2%, respectively; all of the coagulase-negative staphylococci were susceptible to vancomycin. The strains isolated from central venous catheters were identified by a conventional method and the API Staph system. The 27 coagulase-negative Staphylococcus strains were identified as: S. saprophyticus (3.7%), S. xylosus (7.4%), S. haemolyticus (14.8%), S. epidermidis (37.0%), S. warneri (14.8%), S. lugdunensis (7.4%), S. hominis (7.4%), S. schleiferi (3.7%) and S. chromogenes (3.7%). It can be concluded that in the most of the coagulase-negative Staphylococcus species there was an association between slime production, the nosocomial origin of the strains and reduced sensitivity to the antibiotics, suggesting a pathogenic potential in the hospital environment.

In vitro antimicrobial efficiency of a mouthwash containing triclosan/gantrez and sodium bicarbonate

Several antiseptic substances have been used as adjuncts to routine mechanical procedures of oral hygiene, based on their antimicrobial effects. The objective of this study was to assess in vitro the antimicrobial efficiency of 2 mouthwash containing Triclosan/Gantrez and sodium bicarbonate in comparison to both positive and negative controls. Standard strain samples of Escherichia coli, Pseudomonas aeruginosa, Actinomyces viscosus and Bacillus subtilis were used. Samples of Streptococcus mutans and Gram-negative bacilli were collected from 20 volunteers (10 with a clinically healthy periodontium and 10 presenting biofilm-associated gingivitis). Evaluation of the antimicrobial activity was performed by determining the minimal inhibitory concentration (MIC). The results indicated that the test solution inhibited the growth of both Gram-negative and Gram-positive microorganisms from the volunteers' saliva as well as that of the standard strains at the MIC dilution of 1:20, whereas the MIC dilution of 0.12% chlorhexidine against the same bacteria was 1:80. Thus, even though the tested mouthrinse solution presented an in-vitro antimicrobial activity superior to that of a placebo, it was inferior to that of chlorhexidine.

Estudo da reatividade cruzada em amostras de soro de cães positivos para Leishmania sp., Babesia canis e Ehrlichia canis, pelo ensaio imunoenzimático indireto e pela reação de imunofluorescência indireta

To verify the presence of cross-reaction among leishmaniosis, ehrlichiosis and babesiosis in serological diagnostics used in human visceral leishmaniasis control programs, serum samples from leishmaniasis endemic and non-endemic areas were collected and tested by Indirect Fluorescent Antibody (IFAT) and Enzyme-linked immunosorbent assay (ELISA). All serum samples from endemic areas were positive for Leishmania sp., by ELISA and IFAT, 51% positive for Babesia canis and 43% for Ehrlichia canis by IFAT. None of the serum samples from non-endemic areas were positive for Leishmania sp., by IFAT, but 67% were positive for B. canis and 78% for E. canis using the same test. When tested by ELISA for Leishmania sp., four samples from non-endemic area were positive. These dogs were then located and no clinical signs, parasites or antibody was detected in new tests for a six month period. Only one of these 4 samples was positive for B. canis by IFAT and ELISA and three for E. canis by IFAT. The results of the work suggest a co-infection in the endemic area and no serological cross-reaction among these parasites by IFAT and ELISA.

Usefulness of Mycobacterium tuberculosis molecular typing in a tuberculosis low-endemic agro-industrial setting of Brazil

To highlight the transmission and major phylogenetic clades of Mycobacterium tuberculosis, a retrospective study was carried out at two health facilities in a small agro-industrial area in São Paulo, Brazil, that has a low tuberculosis incidence rate. IS6110-RFLP and spoligotyping were performed on the isolates, with the former revealing that 31.3% (35/112) of strains were clustered. Epidemiological links were found in 16 of the 35 clustered patients and were associated with transmission among patients living in public housing. Spoligotyping grouped 62.8% of the strains. The T genetic family predominated among the isolates. Of interest is that five strains had a pattern characteristic of African or Asian origin (ST535), and two others were of the rare localized type ST1888 (BRA, VEN). In addition, three new types-1889, 1890, and 1891-were identified. Spoligotyping showed that some ST may be circulating to or from Brazil, and RFLP revealed ongoing transmission in inadequately ventilated public-housing buildings. This may point to a failure in tuberculosis control policy.

Modulation of the pharmacological effects of enzymatically-active PLA 2 by BTL-2, an isolectin isolated from the Bryothamnion triquetrum red alga

Background. An interaction between lectins from marine algae and PLA 2 from rattlesnake was suggested some years ago. We, herein, studied the effects elicited by a small isolectin (BTL-2), isolated from Bryothamnion triquetrum, on the pharmacological and biological activities of a PLA 2 isolated from rattlesnake venom (Crotalus durissus cascavella), to better understand the enzymatic and pharmacological mechanisms of the PLA 2 and its complex. Results. This PLA2 consisted of 122 amino acids (approximate molecular mass of 14 kDa), its pI was estimated to be 8.3, and its amino acid sequence shared a high degree of similarity with that of other neurotoxic and enzymatically-active PLA2s. BTL-2 had a molecular mass estimated in approximately 9 kDa and was characterized as a basic protein. In addition, BTL-2 did not exhibit any enzymatic activity. The PLA2 and BTL-2 formed a stable heterodimer with a molecular mass of approximately 24-26 kDa, estimated by molecular exclusion HPLC. In the presence of BTL-2, we observed a significant increase in PLA2 activity, 23% higher than that of PLA2 alone. BTL-2 demonstrated an inhibition of 98% in the growth of the Gram-positive bacterial strain, Clavibacter michiganensis michiganensis (Cmm), but only 9.8% inhibition of the Gram-negative bacterial strain, Xanthomonas axonopodis pv passiflorae (Xap). PLA2 decreased bacterial growth by 27.3% and 98.5% for Xap and Cmm, respectively, while incubating these two proteins with PLA2-BTL-2 inhibited their growths by 36.2% for Xap and 98.5% for Cmm. PLA2 significantly induced platelet aggregation in washed platelets, whereas BTL-2 did not induce significant platelet aggregation in any assay. However, BTL-2 significantly inhibited platelet aggregation induced by PLA2. In addition, PLA 2 exhibited strong oedematogenic activity, which was decreased in the presence of BTL-2. BTL-2 alone did not induce oedema and did not decrease or abolish the oedema induced by the 48/80 compound. Conclusion. The unexpected results observed for the PLA2-BTL-2 complex strongly suggest that the pharmacological activity of this PLA2 is not solely dependent on the presence of enzymatic activity, and that other pharmacological regions may also be involved. In addition, we describe for the first time an interaction between two different molecules, which form a stable complex with significant changes in their original biological action. This opens new possibilities for understanding the function and action of crude venom, an extremely complex mixture of different molecules. © 2008 Oliveira et al; licensee BioMed Central Ltd.

Efeito da lavagem peritoneal com bupivacaína na sobrevida de ratos com peritonite fecal

BACKGROUND AND OBJECTIVES: Based on the knowledge of the anti-inflammatory and anti-bacterial actions of local anesthetics (LA), the objective of this study was to determine the effects of peritoneal lavage with bupivacaine on survival of mice with fecal peritonitis. METHODS: Forty-eight Wistar mice, weighing between 300 and 330 g (311.45 ± 9.67 g), undergoing laparotomy 6 hours after induction of peritonitis were randomly divided in 4 groups: 1 - Control, without treatment (n = 12); 2 - Drying of the abdominal cavity (n = 12); 3 - Lavage with 3 mL NS and posterior drying of the abdominal cavity (n = 12); and 4 - Lavage with 8 mg.kg -1 (± 0.5 mL) of 0.5% bupivacaine added to 2.5 mL of NS followed by drying out of the abdominal cavity (n = 12). Animals that died underwent necropsy and the time of death was recorded. Surviving animals were killed on the 11 th postoperative day and underwent necropsy. RESULTS: Group 1 presented a 100% mortality rate in 52 hours, 100% mortality rate in Group 2 in 126 hours, and Group 3 presented a 50% mortality rate in 50 hours. Animals in Group 4 survived. Survival on the 11 th day was greater in groups 3 and 4 than in Groups 1 and 2 (p < 0.001) and greater in Group 4 than in Group 3 (p < 0.01). CONCLUSIONS: Peritoneal lavage with a solution of bupivacaine diluted in NS was effective in preventing death for 11 days in 100% of animals with fecal peritonitis. © Sociedade Brasileira de Anestesiologia, 2008.

Panoramic view of the occurrence of Yersinia species other than Y. pestis in Brazil

Data on the occurrence of Yersinia species, other than Y. pestis in Brazil are presented. Over the past 40 years, 767 Yersinia strains have been identified and typed by the National Reference Center on Yersinia spp. other than Y. pestis, using the classical biochemical tests for species characterization. The strains were further classified into biotypes, serotypes and phagetypes when pertinent. These tests led to the identification of Yersinia cultures belonging to the species Y. enterocolitica, Y. pseudotuberculosis, Y. intermedia, Y. frederiksenii and Y. kristensenii. Six isolates could not be classified in any of the known Yersinia species and for this reason were defined as Non-typable (NT). The bio-sero-phagetypes of these strains were diverse. The following species of Yersinia were not identified among the Brazilian strains by the classical phenotypic or biochemical tests: Y. aldovae, Y. rhodei, Y. mollaretti, Y. bercovieri and Y. ruckeri. The Yersinia strains were isolated from clinical material taken from sick and/or healthy humans and animals, from various types of food and from the environment, by investigators of various Institutions localized in different cities and regions of Brazil.

Aplicabilidade do TTC para a detecção de bacteriúria

Laboratory tests to detect urinary infection at a lawer cost than uroculture are necessary. Triphenyl tetrazolium chloride was evaluated in parallel with uroculture in 342 urine samples, for the detection of significant bacteriuria. The results showed that the test has high sensitivity (91.3%) but low specificity (64.3%), and negative predictive value 99.0%. Although the test cannot replace uroculture as a diagnostic method, it can be recommended for preliminary screening, and could eliminate the need for uroculture of negative samples.