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.

New insights into telomeric DNA sequence (TTAGGG)n location in bat chromosomes

Molossidae species, Cynomops abrasus (2n = 34, fundamental number, FN = 64), Eumops auripendulus (2n = 42, FN = 62), Molossus rufus (2n = 48, FN = 64), Molossops temminckii (2n = 48, FN = 64), and Nyctinomops laticaudatus (2n = 48, FN = 64), and Phyllostomidae species, Phyllostomus discolor (2n = 32, FN = 60), have karyotypes with different chromosome and fundamental numbers, different localization of constitutive heterochromatin, and different numbers and location of nucleolar organizer regions (NORs). Fluorescence in situ hybridization with a human probe of the telomeric sequence (TTAGGG)n produced fluorescent signals in telomeric regions of the six bat species' chromosomes; in E. auripendulus, pericentromeric signals were also observed in the acrocentric and subtelocentric chromosomes. A relationship between telomeric sequences and NORs, and between telomeric sequences and constitutive heterochromatin was detected in chromosomes bearing NORs in C. abrasus, M. temminckii, N. laticaudatus, and P. discolor. No interstitial signal was observed in the meta- or submetacentric chromosomes of these species. ©FUNPEC-RP.

Modeling for performance analysis of electromagnetic zero-sequence suppressor

The purpose of this work is to present a frequency domain model to demonstrate the operation of an electromagnetic arrangement for controlling the injection of zero-sequence currents in the electrical system. Considering the diversity of sequential distribution of harmonic components of a current, the device proposed can be used in the process of mitigation of zero-sequence components. This device, here called electromagnetic suppressor, consists of a blocker and filter both electromagnetic, whose joint operation can provide paths of high and low impedances that can be conveniently adjusted in order to search for a desired performance. This study presents physical considerations, mathematical modeling and computer simulations that clearly demonstrate the viability of this application as a more viable alternative in the conception of filtering systems. The performance analysis is based on the frequency response of harmonic transmittances. The efficacy of this technique in direct actions to maximize the harmonic mitigation process is demonstrated. ©2010 IEEE.

Experimental analysis and adherence test of the electromagnetic zero-sequence blocking model

This paper reveals new contributions to the analysis and development of mitigating harmonic distortion devices. Considering the variety of sequential distribution of harmonic current, in the use of passive filters, one can point out the electromagnetic blocking device, which have received particular attention due to its robustness and low cost of installation. In this context, aiming the evaluation of the reliability of the results obtained through mathematical modeling, experimental tests are carried out using a low-power prototype, highlighting particular aspects related to its function as a zero-sequence harmonic blocking. © 2011 IEEE.

Experimental analysis of an electromagnetic zero-sequence suppressor

The intent of this paper is to present contributions focused on the analysis and development of harmonic attenuator devices. Among these, highlights here the so-called electromagnetic zero-sequence suppressor. This arrangement consists of a filter and a blocker, both electromagnetic, whose combined operation provides paths for low and high impedance, respectively, which can be conveniently adjusted to the desired performance. In this context, here are present results related to experimental studies that show the behavior of the equipment in front of different operating conditions. The tests were performed on a low-power prototype (1kVA/220V) and the analysis results show the main motivator aspects for the use of these devices. © 2012 IEEE.

An assessment of a sequence of yoga exercises to patients with arterial hypertension

This quasi-experimental study describes the effects of a yoga sequence following hemodynamic and biochemical parameters in patients with hypertension. Thirty-three volunteers participated in the study (control = 16 and yoga = 17) for four months. Blood pressure measurements, cardiac and respiratory rate were collected monthly, while the biochemical profile was taken at the beginning and end of the program. To analyze the data, Student's t test and repeated measures analyses were performed. The yoga group showed a significant reduction of systolic blood pressure, heart and respiratory rate (p < 0.05). As for the biochemical profile, the yoga group showed correlation coefficients between initial values and final responses greater than the control of fasting glucose, total cholesterol, LDL-cholesterol and triglycerides. The elaborated sequence practice promoted significant cardiovascular and metabolic benefits. The yoga exercises performed in the proposed sequence constitute complementary non-pharmacological control of blood pressure in patients with hypertension. © 2012 Elsevier Ltd.

Detection of Paracoccidioides spp. in environmental aerosol samples

Taking into account that paracoccidioidomycosis infection occurs by inhalation of the asexual conidia produced by Paracoccidioides spp. in its saprobic phase, this work presents the collection of aerosol samples as an option for environmental detection of this pathogen, by positioning a cyclonic air sampler at the entrance of armadillo burrows. Methods included direct culture, extinction technique culture and Nested PCR of the rRNA coding sequence, comprising the ITS1-5.8S-ITS2 region. In addition, we evaluated one armadillo (Dasypus novemcinctus) as a positive control for the studied area. Although the pathogen could not be isolated by the culturing strategies, the aerosol sampling associated with molecular detection through Nested PCR proved the best method for discovering Paracoccidioides spp. in the environment. Most of the ITS sequences obtained in this investigation proved to be highly similar with the homologous sequences of Paracoccidioides lutzii from the GenBank database, suggesting that this Paracoccidioides species may not be exclusive to mid-western Brazil as proposed so far. © 2013 ISHAM.

Filamentous fungi from the Atlantic marine sponge Dragmacidon reticulatum

Dragmacidon reticulatum is a marine sponge of wide occurrence in the Eastern and Western Atlantic. Little is known about D. reticulatum fungal diversity. Filamentous fungi recovered from D. reticulatum were assessed in the present study using a polyphasic taxonomic approach, including classical morphology, molecular biology and MALDI-TOF ICMS. Ninety-eight fungal strains were isolated from two D. reticulatum samples by using six different culture media, which were identified up to the genus level. Sixty-four distinct fungal ribotypes were obtained, distributed among twenty-four different genera belonging to the Ascomycota and Zygomycota. Representatives of Penicillium and Trichoderma were the most diverse and abundant fungi isolated. Amongst Penicillium spp. three isolates belonged to the same ribotype can be considered as a putative new species. Data derived from the present study highlight the importance of using a polyphasic approach to get an accurate identification in order to structure a reliable culture collection. © 2012 Springer-Verlag Berlin Heidelberg.

Estimation of spontaneous blinking main sequence in normal subjects and patients with Graves' upper eyelid retraction

Purpose. We quantified the main sequence of spontaneous blinks in normal subjects and Graves' disease patients with upper eyelid retraction using a nonlinear and two linear models, and examined the variability of the main sequence estimated with standard linear regression for 10-minute periods of time. Methods. A total of 20 normal subjects and 12 patients had their spontaneous blinking measured with the magnetic search coil technique when watching a video during one hour. The main sequence was estimated with a power-law function, and with standard and trough the origin linear regressions. Repeated measurements ANOVA was used to test the mean sequence stability of 10-minute bins measured with standard linear regression. Results. In 95% of the sample the correlation coefficients of the main sequence ranged from 0.60 to 0.94. Homoscedasticity of the peak velocity was not verified in 20% of the subjects and 25% of the patients. The power-law function provided the best main sequence fitting for subjects and patients. The mean sequence of 10-minute bins measured with standard linear regression did not differ from the one-hour period value. For the entire period of observation and the slope obtained by standard linear regression, the main sequence of the patients was reduced significantly compared to the normal subjects. Conclusions. Standard linear regression is a valid and stable approximation for estimating the main sequence of spontaneous blinking. However, the basic assumptions of the linear regression model should be examined on an individual basis. The maximum velocity of large blinks is slower in Graves' disease patients than in normal subjects. © 2013 The Association for Research in Vision and Ophthalmology, Inc.

Cooperative RNA Polymerase Molecules Behavior on a Stochastic Sequence-Dependent Model for Transcription Elongation

The transcription process is crucial to life and the enzyme RNA polymerase (RNAP) is the major component of the transcription machinery. The development of single-molecule techniques, such as magnetic and optical tweezers, atomic-force microscopy and single-molecule fluorescence, increased our understanding of the transcription process and complements traditional biochemical studies. Based on these studies, theoretical models have been proposed to explain and predict the kinetics of the RNAP during the polymerization, highlighting the results achieved by models based on the thermodynamic stability of the transcription elongation complex. However, experiments showed that if more than one RNAP initiates from the same promoter, the transcription behavior slightly changes and new phenomenona are observed. We proposed and implemented a theoretical model that considers collisions between RNAPs and predicts their cooperative behavior during multi-round transcription generalizing the Bai et al. stochastic sequence-dependent model. In our approach, collisions between elongating enzymes modify their transcription rate values. We performed the simulations in Mathematica® and compared the results of the single and the multiple-molecule transcription with experimental results and other theoretical models. Our multi-round approach can recover several expected behaviors, showing that the transcription process for the studied sequences can be accelerated up to 48% when collisions are allowed: the dwell times on pause sites are reduced as well as the distance that the RNAPs backtracked from backtracking sites. © 2013 Costa et al.

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

The pioneering use of ISSR (Inter Simple Sequence Repeat) in Neotropical anurans: Preliminary assessment of genetic diversity in populations of Physalaemus cuvieri (Amphibia, Leiuperidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The population genetic structure of Rhizoctonia solani AG-3PT from potato in the Colombian Andes

The soilborne fungus Rhizoctonia solani anastomosis group 3 (AG-3PT) is a globally important potato pathogen. However, little is known about the population genetic processes affecting field populations of R. solani AG-3PT, especially in the South American Colombian Andes, which is near the center of diversity of the two most common groups of cultivated potato, Solanum tuberosum and S. phureja. We analyzed the genetic structure of 15 populations of R. solani AG-3PT infecting potato in Colombia using 11 simple-sequence repeat (SSR) markers. In total, 288 different multilocus genotypes were identified among 349 fungal isolates. Clonal fractions within field populations were 7 to 33%. R ST statistics indicated a very low level of population differentiation overall, consistent with high contemporary gene flow, though moderate differentiation was found for the most distant southern populations. Genotype flow was also detected, with the most common genotype found widely distributed among field populations. All populations showed evidence of a mixed reproductive mode, including both asexual and sexual reproduction, but two populations displayed evidence of inbreeding. © 2013 The American Phytopathological Society.

Using the bottom-up and outside-in sequence for panfacial fracture management: Does it provide a clinical significance?

Panfacial fractures usually refer to simultaneous facial fractures, which affect the upper, middle, and lower thirds of the face. The management of panfacial fracture is complex because of the lack of reliable landmarks. Literature has shown many approaches for management of panfacial fractures. Every segment of bone has a precise function in the repair. Therefore, the bottom-up and outside-in sequence is the most widely used approach in the management of panfacial fractures. These facial fractures present remarkable challenges for both experienced and inexperienced surgeons. This article aimed to report a case of a panfacial fracture (mandibular condylar and symphysis fractures associated with an atypical Le Fort III fracture) in a 48-year-old man. The patient was successfully treated using bottom-up and outside-in sequence by accessing all facial injuries. Postoperatively, radiograph examination revealed good reduction and fixation of titanium plates, and physical examination revealed good functional and esthetic outcomes. Copyright © 2013 by Mutaz B. Habal, MD.

Multilocus molecular phylogeny of Gasteropelecidae (Ostariophysi: Characiformes) reveals the existence of an unsuspected diversity

The characiform family Gasteropelecidae, the so-called freshwater hatchetfishes, is comprised of three genera and nine species found in Panama and all South American countries except Chile. Our goal was to investigate the molecular characteristics, phylogenetic relationships among the species and genera of Gasteropelecidae and phylogenetic relationships between the Gasteropelecidae family with other Characiformes. DNA fragments from two mitochondrial (16S rRNA and Cytochrome B) and three nuclear genes (Rag1, Rag2 and Myh6) were sequenced. Our results corroborate the morphology-based hypothesized monophyly of the Gasteropelecidae family and most of the relationships among its genera. However, the genus Gasteropelecus is polyphyletic because G. maculatus is placed as the sister group to all other gasteropelecids, whereas G. sternicla is more closely related to species of Carnegiella. Similarly, the species Carnegiella strigata is not monophyletic, which suggests that the family needs a taxonomic review. Moreover, the species Thoracocharax stellatus was composed by four distinct lineages suggesting the this species may represents a species complex. © 2013 Elsevier Inc.