Taenia solium and Taenia saginata: Identification of sequence characterized amplified region (SCAR) markers

Cysticercosis is one of the most important zoonosis, not only because of the effects on animal health and its economic consequences, but also due to the serious danger it poses to humans. The two main parasites involved in the taeniasis-cysticercosis complex in Brazil are Taenia saginata and Taenia solium. Differentiating between these two parasites is important both for disease control and for epidemiological studies. The purpose of this work was to identify genetic markers that could be used to differentiate these parasites. Out of 120 oligonucleotide decamers tested in random amplified polymorphic DNA (RAPD) assays, 107 were shown to discriminate between the two species of Taenia. Twenty-one DNA fragments that were specific for each species of Taenia were chosen for DNA cloning and sequencing. Seven RAPD markers were converted into sequence characterized amplified region (SCAR) markers with two specific for T. saginata and five specific for T. solium as shown by agarose gel electrophoresis. These markers were developed as potential tools to differentiate T. solium from T. saginata in epidemiological studies. © 2007 Elsevier Inc. All rights reserved.

Effect of mating delay in the ovary of Apis mellifera L. (Hymenoptera, Apidae) queens: Histological aspects

This paper reports the effect of mating delay on the queen Apis mellifera ovaries based on a light microscopy analysis. Soon after a queen emerges from the brood cell, oocytes start to differentiate in the ovaries, but if mating does not occur at the correct age (about 6 days after emergence) cell degeneration begins. Ovaries of 15-day-old virgin queens show extensive disorganization with cell death affecting all types of ovariole cells. Types of cell death and the possible causes are discussed.

Phenotypic characterization of Candida spp. isolates from chronic periodontitis patients

Aim: Several typing methods for Candida spp. have been suggested in the literature in order to distinguish isolates for studies about the virulence or infection routes of these microorganisms and, in particular, for epidemiological purposes. The aim of this study was to establish a comparison between the phenotypic profile of oral Candida isolates from periodontitis patients and control individuals. Methods: The morphotyping and biotyping of 35 C. albicans isolates obtained from chronic periodontitis patients and 48 isolates from control individuals were performed. For morphotyping, the isolates were plated on malt extract agar and incubated for 10 days. Sixteen different morphotypes were observed for C. albicans, the most frequently observed being 0000 and 0001. Results: Biotype 0000 (complete absence of fringe) was most prevalent among the isolates obtained from periodontitis patients compared to those from control individuals, with statistical significance. Biotyping revealed 5 different biotypes with higher prevalence of the biotype 357 among the isolates from control and periodontitis groups. Conclusions: The results obtained by biotyping of the isolates did not permit to differentiate a characteristic model related to periodontal disease, whilst the morphotype 0000 was most frequently isolated from periodontitis patients.

Caste differences in Polybia ematiata, a mud-nesting eusocial wasp (hymenoptera: Vespidae), that bear on pre-imaginal determination but not size differences

The Epiponini is a group of social wasps living in polygynic societies. The caste dimorphism varies from incipient to well distinct. Morphometric analyses on twelve body parts and ovarian development of Polybia emaciata Lucas 1879, were used to estimate the level of caste differentiation in seven colonies. Physiologic differences were found. Among queens the ovaries were inseminated and presented as highly developed; in intermediate females, ovaries were slightly developed and uninseminated, and for the workers the ovaries were not developed or inseminated. In addition, the coloration of the mesosoma and metasoma were darker in workers and intermediates. Morphometric analysis evidenced slight morphological differences. Even though body proportions were not enough to discriminate castes, the fifth sternite of the gaster had important characters that were able to differentiate queens from workers and intermediates. We propose that P. emaciata presents a case of pre-imaginal caste determination, which is not strictly based on size.

Testes de vigor para avaliação da qualidade fisiológica de sementesde Sebastiania commersoniana (Baill.) Smith & Downs.

The objective of this work was to evaluate the efficiency of different vigor tests to assess S. commersoniana seeds physiological quality during storage. Therefore, seeds were stored in cloth bags, plastic bags and glass containers for 531 days, both at room temperature and in a cold chamber. Periodically samples were taken and the following tests were conducted: standard germination, germination speed index, first count after germination, seedlings fresh and dry matter, electrical conductivity (imbibitions for 2 and 24 hours) and accelerated aging, in order to monitoring the seeds physiologic quality. Data were submitted to polynomial regression analysis, for each of the combination of packaging/storage condition. The results obtained led to the following considerations: a) the speed germination index and the accelerated aging test permitted a good evaluation of the seed-lots quality; b) the first count of germination did not allow any prediction about seed lots vigor; c) seedling fresh and dry matter did not discriminate among seed lots; d) electrical conductivity test after 2 h imbibitions was not adequate to evaluate the seed lots vigor and e) the electric conductivity test with 24 h imbibitions was more adequate to differentiate among seed lots, but with little efficiency to discriminate among seed lots with intermediate vigor or with low quality differences.

Métodos de avaliação da eficácia dos tratamentos para a dor.

Pain is a subjective condition and, thus, difficult to measure. The best tools to assess pain are the pain evaluation questionnaires, which provide either diagnostic, pain evolution or pain intensity information. To provide information which could help differentiate between nociceptive pain and neuropathic pain is one of the most important functions of these questionnaires. The questionnaires can measure pain intensity, quality of life, or sleep quality. Quality of life and sleep are two really important characteristics to assess the pain impact on patients' life. Pain intensity assessing questionnaires combine physical evaluations with questions, providing information either from the patient sensations or clinical assessment of pain manifestations as well as the underlying biological mechanisms (such as hyperalgesia or allodynia). For example, the Pain Detect questionnaire has two parts: the patient form (intuitive, with pictures and easy understandable) and the physician form. Thus, in this questionnaire, subjective information is provided by the patient and the objective one is provided by the physician. Other pain intensity questionnaires are NPSI, DN4, LANSS or StEP. Quality of life questionnaires are versatile (can be used in different pathologies). These questionnaires include functional self-evaluation questions, and other ones associated to physical and mental health. Two of such quality of life questionnaires are SF-36 and NHP. Sleep evaluation questionnaires include quantitative features such as the number of sleep interruptions, sleep latency or sleep duration as well as qualitative characteristics such as rest sensation, mood and dreams. One of the most used sleep evaluation questionnaires is PSQI, which includes patient questions and bed-partner questions, providing information from two points of view. Copyright 2009 Prous Science, S.A.U. or its licensors. All rights reserved.

Práticas educativas parentais e repertório comportamental infantil: Comparando crianças diferenciadas pelo comportamento

Identifying parental practices contingent to the behavior of children is important to support interventions. This study compares parental disciplinary practices with the behavior of children in a clinical group with behavioral problems (n = 27) and of a group of children without behavioral problems (n = 26). The criterion to assign children in the clinical and non-clinical groups was obtained from the Child Behavior Checklist (CBCL). Information concerning parents-children interaction was obtained through the application of the Parental Educative Social Skills Interview Script (RE-HSE-P), which evaluates positive and negative educational practices, context, behavioral problems and children's social skills. Qualitative and quantitative analyses (Mann Whitney's test) were used in the groups' comparison. The results indicate that behaviors that differentiate between the clinical and non-clinical groups are mainly those related to positive educational practices and children's social skills.

Macroeconomic coordination in Latin America: does it have a future?

Includes bibliography

Use of reverse transcriptase polymerase chain reaction (RT-PCR) in molecular screening of Newcastle disease virus in poultry and free-living bird populations

The aim of this study was to evaluate a simple molecular method of reverse transcriptase polymerase chain reaction (RT-PCR) to differentiate Newcastle disease virus strains according to their pathogenicity, in order to use it in molecular screening of Newcastle disease virus in poultry and free-living bird populations. Specific primers were developed to differentiate LaSota-LS-(vaccine strain) and Sao Joao do Meriti-SJM-strain (highly pathogenic strain). Chickens and pigeons were experimentally vaccinated/infected for an in vivo study to determine virus shedding in feces. Validation of sensitivity and specificity of the primers (SJM and LS) by experimental models used in the present study and results obtained in the molecular analysis of the primers by BLAST made it possible to generalize results. The development of primers that differentiate the level of pathogenicity of NDV stains is very important, mainly in countries where real-time RT-PCR is still not used as a routine test. These primers were able to determine the presence of the agent and to differentiate it according to its pathogenicity. © 2012 Springer Science+Business Media B.V.

Analysis of spermiogenesis like a tool in the study of the triatomines of the Brasiliensis subcomplex

The specific identification and systematic of triatomines have been based fundamentally on morphological observations. These organisms are classified into complexes and specific subcomplexes, principally for morphological parameters and geographical disposition. The use of cytogenetic analyzes has been represented as a tool in systematic and taxonomy of triatomines. Thus, the present work, through the analysis of spermiogenesis, aims to characterize this stage of spermatogenesis in triatomines little studied, and especially to compare it among the species Triatoma lenti and T. sherlocki, to assist in the diagnosis of differentiation of these insects. The presence of the heteropyknotic corpuscle is shown as a diagnostic tool to differentiate T. sherlocki and T. lenti, since it is absent in T. lenti. The analysis of the spermiogenesis in T. sherlocki also allowed us to address morphological differences between elongating cells, which were relatively smaller and more filamentous when compared to T lenti. Furthermore, the flagellum was observed in all stages of cell differentiation and elongation. This structure, which helps in the locomotion of the sperm, is hardly observed in cytogenetic analysis, especially throughout spermiogenesis. Thus, although other comparative approaches should be taken, this paper allowed emphasizing the analysis of spermiogenesis as an important cytotaxonomic tool that assists in the differentiation of morphologically related species, such as T. lenti and T. sherlocki. © 2013 Académie des sciences.

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

Salmonella Pullorum and Salmonella Gallinarum are classified as biovars of Salmonella enterica subsp. enterica serovar Gallinarum. These salmonellae are the causative agents of Pullorum disease and fowl typhoid, respectively, and are widely distributed throughout the world. Although many developed countries have eradicated these diseases from commercial poultry, they are still the cause of significant economic loss in developing countries. When serovar Gallinarum is isolated, it is difficult to immediately differentiate between biovars because they are antigenically identical by serotyping. However, they cause distinct diseases with different epidemiology, and therefore it is important to differentiate them. This may be done biochemically but takes 2 to 3 days. In the present study, S. Pullorum and S. Gallinarum whole genomes were compared, and 1 genomic region of difference, which is part of the ratA gene, was chosen as a molecular marker for a polymerase chain reaction assay to differentiate rapidly between these organisms. In all, 26 strains of S. Gallinarum and 17 S. Pullorum strains were tested and successfully differentiated by the assay. © 2013 The Author(s).

Development of an analytical method for the quantification of pfaffic acid in Brazilian ginseng (Hebanthe eriantha)

Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. © 2013 Elsevier B.V.

Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: An improved approach in cartilage tissue engineering

Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n=5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1×105) were than encapsulated inside 60μl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI. © 2013 Informa UK Ltd.

Elucidation of carbohydrate molecular interaction mechanism of recombinant and native ArtinM

The quartz crystal microbalance (QCM) technique has been applied for monitoring the biorecognition of ArtinM lectins at low horseradish peroxidase glycoprotein (HRP) concentrations, using a simple kinetic model based on Langmuir isotherm in previous work.18 The latter approach was consistent with the data at dilute conditions but it fails to explain the small differences existing in the jArtinM and rArtinM due to ligand binding concentration limit. Here we extend this analysis to differentiate sugar-binding event of recombinant (rArtinM) and native (jArtinM) ArtinM lectins beyond dilute conditions. Equivalently, functionalized quartz crystal microbalance with dissipation monitoring (QCM-D) was used as real-time label-free technique but structural-dependent kinetic features of the interaction were detailed by using combined analysis of mass and dissipation factor variation. The stated kinetic model not only was able to predict the diluted conditions but also allowed to differentiate ArtinM avidities. For instance, it was found that rArtinM avidity is higher than jArtinM avidity whereas their conformational flexibility is lower. Additionally, it was possible to monitor the hydration shell of the binding complex with ArtinM lectins under dynamic conditions. Such information is key in understanding and differentiating protein binding avidity, biological functionality, and kinetics. © 2013 American Chemical Society.

Analysis of inteins in the Candida parapsilosis complex for simple and accurate species identification

Inteins are coding sequences that are transcribed and translated with flanking sequences and then are excised by an autocatalytic process. There are two types of inteins in fungi, mini-inteins and full-length inteins, both of which present a splicing domain containing well-conserved amino acid sequences. Full-length inteins also present a homing endonuclease domain that makes the intein a mobile genetic element. These parasitic genetic elements are located in highly conserved genes and may allow for the differentiation of closely related species of the Candida parapsilosis (psilosis) complex. The correct identification of the three psilosis complex species C. parapsilosis, Candida metapsilosis, and Candida orthopsilosis is very important in the clinical setting for improving antifungal therapy and patient care. In this work, we analyzed inteins that are present in the vacuolar ATPase gene VMA and in the threonyl-tRNA synthetase gene ThrRS in 85 strains of the Candida psilosis complex (46 C. parapsilosis, 17 C. metapsilosis, and 22 C. orthopsilosis). Here, we describe an accessible and accurate technique based on a single PCR that is able to differentiate the psilosis complex based on the VMA intein. Although the ThrRS intein does not distinguish the three species of the psilosis complex by PCR product size, it can differentiate them by sequencing and phylogenetic analysis. Furthermore, this intein is unusually present as both mini- and full-length forms in C. orthopsilosis. Additional population studies should be performed to address whether this represents a common intraspecific variability or the presence of subspecies within C. orthopsilosis. Copyright © 2013, American Society for Microbiology. All Rights Reserved.