Isolation of Mycobacterium spp. in milk from cows suspected or positive to tuberculosis

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

Isolation and HPLC quantitative analysis of flavonoid glycosides from Brazilian beverages (Maytenus ilicifolia and M-aquifolium)

Aqueous infusions of Brazilian Maytenus leaves are used as beverages, foodstuffs, and phytomedicines. Previously, we isolated two new flavonoid tetrasaccharides from the infusion of Maytenus aquifolium leaves that showed antiulcer activity. In this investigation a new flavonoid tetrasaccharide, kaempferol-3-O-alpha -L-rhamnopyranosyl (1-6)-O-[alpha -L-arabinopyranosyl (1 -->3)-O-alpha -L-rhamnopyranosyl (1-2)]-O-beta -D-galactopyranoside (3), was isolated, together with kaempferol tri- and disaccharides and quercetin trisaccharides from the aqueous infusion of Maytenus ilicifolia leaves. All structures were elucidated by ES-MS and NMR spectroscopic methods. The quantitative analysis of the flavonoid glycosides from Maytenus ilicifolia and M. aquifolium has been performed by HPLC.

Isolation and structure elucidation of two new flavonoid glycosides from the infusion of Maytenus aquifolium leaves. Evaluation of the antiulcer activity of the infusion

Droplet countercurrent chromatography and high-performance liquid chromatography fractionation of the aqueous infusion from Maytenus aquifolium Martius leaves afforded two flavonoid tetrasaccharides: quercetin 3-O-alpha-L-rhamnopyranosyl(1-->6)-O-[beta-D-glucopyranosyl(1-->3)-O-alpha-L-rhamnopranosyl( 1-->2)-O-beta-D-galactopyranoside and kaempferol 3-O-alpha-L-rhamnopyranosyl(1-->6)-O-[beta-D-glucopyranosyl( 1-->3)-O-alpha-L-rhamnopyranosy(1-->2-2)-O-beta-D-galactopyranoside. All structures were elucidated by spectroscopic methods. Pharmacological essays of the infusion showed antiulcer activity in rats.

In vitro evaluation of root canal preparation using oscillatory and rotary systems in flattened root canals

Objective: the aim of this study was to evaluate the biomechanical preparation of flattened root canals using the following systems: Endo-Eze AET stainless steel oscillatory instruments (Ultradent) and RaCe rotary NiTi instruments (FKG Dentaire). Materials and Methods: Twenty extracted human mandibular incisors were randomly assigned to two groups: Group I Instrumentation with oscillatory Endo-Eze AET files (oscillatory technique); Group 2 - Instrumentation with rotary NiTi RaCe files (rotary technique). The teeth were decoronated, had their apices and coronal openings sealed with sticky wax and were embedded in crystal-clear orthophtalic polyester resin. The roots were sectioned transversally with diamond discs at 10 mm (middle third) and 5 mm (apical third) from the apex and the segments were reassembled for instrumentation. The sections were photographed before and after root canal instrumentation and evaluated with respect to whether the original root canal shape was modified by instrumentation. To evaluate the differences in the root canal shape before and after biomechanical preparation, scores were given regarding the instruments touch on the intracanal walls. Results: In middle third of the root canals instrumented with the rotary system, there was a change in the original canal anatomy (p < 0.05), with formation of a protuberance in the mesiodistal direction. This protuberance did not occur when the oscillatory instrumentation was used. The oscillatory system had better results in the middle and apical thirds as evaluated by Dunn's multiple-comparison test (p > 0.05). Conclusion: Under the tested conditions, Endo-Eze oscillatory system yielded the instrumentation of all flattened oot canal walls, maintaining the canal original shape throughout the biomechanical preparation, and was more effective than RaCe rotary system.

Studies on the constituents of a Brazilian folk infusion. Isolation and structure elucidation of new triterpene saponins from Ilex amara leaves

The isolation of three new triterpene saponins 3beta-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-2-O-acetylara-binopyranosylolean-12-en-28-oic acid 28-O-beta-D-glucopyranosyl ester (2), 3beta-O-beta-D-glucopyranosyl-(1-->2)-alpha-L-O-arabinopyranosylurs-12-en-28-oic acid (3), and 3beta-O-beta-D-glucopyranosyl-(1-->2)-beta-D-O-galactopyranosylurs-12-en-28-oic acid (4) together with five known saponins and one flavonoid glycoside from the aqueous infusion of flex amara (Vellozo) Loes. leaves is reported. All structures were elucidated by spectroscopic methods, including the concerted application of one-dimensional (H-1, TOCSY, C-13, and C-13 DEPT NMR) and two-dimensional NMR techniques (DQF-COSY, HSQC, and HMBC).

Molecular fingerprinting methods for the discrimination between C-albicans and C-dubliniensis

Opportunistic fungal pathogens are becoming increasingly important causes of both community-acquired and nosocomial infections. The most important fungal pathogens are yeast species belonging to the genus Candida. These species show differences in levels of resistance to antifungal agents and mortality. Consequently, it is important to correctly identify the causative organism to the species level. Identification of Candida dubliniensis in particular remains problematic because of the high degree of phenotypic similarity between this species and Candida albicans. However, as the differences between both are most pronounced at the genetic level, several studies have been conducted in order to provide a specific and rapid identification fingerprinting molecular test. In most candidal infectious, no single DNA fingerprinting technique has evolved as a dominant method, and each method has its advantages, disadvantages and limitations. Moreover, the current challenge of these techniques is to compile standardized patterns in a database for interlaboratory use and future reference. This review provides an overview of most common molecular fingerprinting techniques currently available for discrimination of C. albicans and C. dubliniensis.

Progress on holographic techniques to measure real-time phase and amplitude gratings in photosensitive materials

In this paper we present and demonstrate a technique that allows simultaneous and independent measurement of small changes in the refractive index and the absorption coefficient produced in photosensitive materials during holographic exposure. The technique is based on the synchronous detection of two-wave mixing signals in both directions of the transmitted interfering beams. By processing both signals it is possible to separate the diffraction contributions of the refractive index from the absorption coefficient and simultaneously stabilize the incident fringe pattern. The demonstration of this technique is undertaken by following the temporal evolution of the phase and amplitude modulations in photoresist films. To check the ability of the technique to perform numeric evaluations, for a positive photoresist the changes in the optical constants were measured and compared with those obtained using independent methods.

Effects of Nd : YAG laser irradiation on root canal dentin wall: A scanning electron microscopic study

Objective: the purpose of this study was to evaluate, by scanning electron microscopy (SEM), the effects of Nd:YAG laser irradiation applied perpendicular or parallel to the root canal dentin wall. Methods: Thirty human teeth were divided into two groups: Group A (20 roots), laser application with circular movements, parallel to the dentin root surface; and Group B (10 roots), roots cut longitudinally and laser applied perpendicular to the root surface. Group A was subdivided into A1 (10 roots), laser application with 100 mJ, 15 Hz and 1.5 W; and A2 (10 roots) with 160 mJ, 15 Hz, and 2.4 W. Group B was subdivided into B1 (10 hemisections) and B2 (10 hemi-sections) with parameters similar to A I and A2. Four applications of 7-sec duration were performed, with a total exposure of 28 sec. SEM evaluations were made in the cervical, middle, and apical thirds, with 500X and 2000X magnifications. Morphological changes scores were attributed, and the results were submitted to Kruskal Wallis statistical test (5%). Results: Significant statistical differences were found between groups A and B (p = 0.001). In groups A1 and A2, few areas of dentin melting were observed. In groups B1 and B2, areas of melting dentin covering dentin surface were observed. Conclusions: It was concluded that intracanal laser application with circular movements (parallel to the surface) produces limited morphological changes in root canal dentin wall.

Phenotypic methods and commercial systems for the discrimination between C-albicans and C-dubliniensis

Candida dubliniensis is a recently described Candida species associated with oral candidosis that exhibits a high degree of phenotypic similarity to Candida albicans. However, these species show differences in levels of resistance to antimycotic agents and ability to cause infections. Therefore, accurate clinical identification of C. dubliniensis and C. albicans species is important in order to treat oral candidal infections. Phenotypic identification methods are easy-to-use procedures for routine discrimination of oral isolates in the clinical microbiology laboratory. However, C. dubliniensis may be so far underreported in clinical samples because most currently used identification methods fail to recognize this yeast. Phenotypic methods depend on growth temperature, carbon source assimilation, chlamydospore and hyphal growth production, positive or negative growth on special media and intracellular enzyme production, among others. In this review, some phenotypic methods are presented with a special emphasis on the discrimination of C. dubliniensis and C. albicans.

An intelligent system to real time rainfall prediction using radar data

This work presents a new approach for rainfall measurements making use of weather radar data for real time application to the radar systems operated by institute of Meteorological Research (IPMET) - UNESP - Bauru - SP-Brazil. Several real time adjustment techniques has been presented being most of them based on surface rain-gauge network. However, some of these methods do not regard the effect of the integration area, time integration and distance rainfall-radar. In this paper, artificial neural networks have been applied for generate a radar reflectivity-rain relationships which regard all effects described above. To evaluate prediction procedure, cross validation was performed using data from IPMET weather Doppler radar and rain-gauge network under the radar umbrella. The preliminary results were acceptable for rainfalls prediction. The small errors observed result from the spatial density and the time resolution of the rain-gauges networks used to calibrate the radar.

Comparative studies on non-convex optimization methods for transmission network expansion planning

We have investigated and extensively tested three families of non-convex optimization approaches for solving the transmission network expansion planning problem: simulated annealing (SA), genetic algorithms (GA), and tabu search algorithms (TS). The paper compares the main features of the three approaches and presents an integrated view of these methodologies. A hybrid approach is then proposed which presents performances which are far better than the ones obtained with any of these approaches individually. Results obtained in tests performed with large scale real-life networks are summarized.

Healing of dehiscence defects following root surface demineralization with tetracycline: A histologic study in monkeys

Background: the purpose of this study was to histologically evaluate the healing of experimental dehiscence defects after surface demineralization with tetracycline hydrochloride.Methods: Six adult male monkeys (Cebus apella) were used in this study. Dehiscence defects were surgically created on the buccal aspect of the mandibular lateral incisors in all animals. The root surfaces were debrided and planed. In a split-mouth design, a 10% tetracycline hydrochloride solution was applied to one tooth for 4 minutes (T group), followed by irrigation with saline. The contralateral tooth served as a control (C group). The flaps were repositioned and sutured. The animals were sacrificed at 6 months postoperatively and histological sections were processed. Computer-assisted histomorphometric analysis was used to evaluate the formation of new cementum, new bone, new connective tissue attachment, and length of the epithelium (junctional and sulcular).Results: Bone regeneration was similar in both groups (1.5 +/- 0.3 mm for the T group and 1.5 +/- 0.6 mm for the C group). The C group showed more new cementum than the T group (2.3 +/- 0.3 mm versus 2.2 +/- 0.3 mm) as well as a longer epithelium (1.0 +/- 0.3 mm versus 0.9 +/- 0.2 mm). The T group presented more new connective tissue attachment (3.1 +/- 0.2 mm) than the C group (2.9 +/- 0.6 mm). However, no statistically significant differences were detected between the two groups.Conclusions: the amount of new attachment was similar in both groups. Root conditioning with 10% tetracycline solution did not produce any additional new attachment in comparison to the controls.

Periodontal status in smokers and never-smokers: Clinical findings and real-time polymerase chain reaction quantification of putative periodontal pathogens

Background: Smoking is a well-known risk factor for destructive periodontal disease, but its relationship with periodontal status and subgingival microbiota remains unclear. Inherent limitations of microbiological methods previously used may partly explain these mixed results, and real-time polymerase chain reaction (PCR) has been presented as a valid alternative. The aim of the present study was to investigate the clinical condition and microbiological profile of patients with chronic periodontitis as related to the habit of smoking.Methods: Fifty patients (33 to 59 years old), 25 smokers and 25 never-smokers, constituted the sample. The visible plaque index (VPI), gingival bleeding index (GBI), bleeding on probing (BOP), periodontal probing depth (PD), clinical attachment loss (CAL), and gingival crevicular fluid (GCF) volume were recorded. Real-time PCR quantified Porphyromonas gingivalis, Micromonas micros, Dialister pneumosintes, Actinobacillus actinomycetemcomitans and total bacteria in subgingival samples.Results: Smokers and never-smokers showed similar values for VPI, GBI, and BOP. Smokers had deeper PD in buccal/lingual sites and higher CAL independently of the tooth surface. The GCF volume was smaller in smokers, independent of the PD. Similar amounts of total bacteria and P. gingivalis were observed for both groups. Significantly higher numbers of D. pneumosintes and M. micros were present in smokers and associated with moderate and deep pockets. When heavy smokers were considered, higher counts of total bacteria, M. micros, and D. pneumosintes were observed.Conclusions: Smoking seems to have a detrimental impact on the periodontal status and microbiological profile of patients with periodontitis. Compared to never-smokers, smokers had deeper pockets, greater periodontal destruction, and higher counts of some putative periodontal pathogens.

Analysis of Apical Root Transportation Associated with Protaper Universal F3 and F4 Instruments by Using Digital Subtraction Radiography

Introduction: The aim of this study was to assess the occurrence of apical root transportation after the use of Pro Taper Universal rotary files sizes 3 (F3) and 4 (F4). Methods: Instruments were worked to the apex of the original canal, always by the same operator. Digital subtraction radiography images were produced in buccolingual and mesiodistal projections. A total of 25 radiographs were taken from root canals of human maxillary first molars with curvatures varying from 23-31 degrees. Quantitative data were analyzed by intraclass correlation coefficient and Wilcoxon nonparametric test (P = .05). Results: Buccolingual images revealed a significantly higher degree of apical transportation associated with F4 instruments when compared with F3 instruments in relation to the original canal (Wilcoxon test, P = .007). No significant difference was observed in mesiodistal images (P = .492). Conclusions: F3 instruments should be used with care in curved canals, and F4 instruments should be avoided in apical third preparation of curved canals. (J Endod 2010;36:1052-1055)

New methods to quantify NH3 volatilization from fertilized surface soil with urea

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