The Mycobacterium tuberculosis Rv2540c DNA sequence encodes a bifunctional chorismate synthase

Background. The emergence of multi- and extensively-drug resistant Mycobacterium tuberculosis strains has created an urgent need for new agents to treat tuberculosis (TB). The enzymes of shikimate pathway are attractive targets to the development of antitubercular agents because it is essential for M. tuberculosis and is absent from humans. Chorismate synthase (CS) is the seventh enzyme of this route and catalyzes the NADH- and FMN-dependent synthesis of chorismate, a precursor of aromatic amino acids, naphthoquinones, menaquinones, and mycobactins. Although the M. tuberculosis Rv2540c (aroF) sequence has been annotated to encode a chorismate synthase, there has been no report on its correct assignment and functional characterization of its protein product. Results. In the present work, we describe DNA amplification of aroF-encoded CS from M. tuberculosis (MtCS), molecular cloning, protein expression, and purification to homogeneity. N-terminal amino acid sequencing, mass spectrometry and gel filtration chromatography were employed to determine identity, subunit molecular weight and oligomeric state in solution of homogeneous recombinant MtCS. The bifunctionality of MtCS was determined by measurements of both chorismate synthase and NADH:FMN oxidoreductase activities. The flavin reductase activity was characterized, showing the existence of a complex between FMN ox and MtCS. FMNox and NADH equilibrium binding was measured. Primary deuterium, solvent and multiple kinetic isotope effects are described and suggest distinct steps for hydride and proton transfers, with the former being more rate-limiting. Conclusion. This is the first report showing that a bacterial CS is bifunctional. Primary deuterium kinetic isotope effects show that C4-proS hydrogen is being transferred during the reduction of FMNox by NADH and that hydride transfer contributes significantly to the rate-limiting step of FMN reduction reaction. Solvent kinetic isotope effects and proton inventory results indicate that proton transfer from solvent partially limits the rate of FMN reduction and that a single proton transfer gives rise to the observed solvent isotope effect. Multiple isotope effects suggest a stepwise mechanism for the reduction of FMNox. The results on enzyme kinetics described here provide evidence for the mode of action of MtCS and should thus pave the way for the rational design of antitubercular agents. © 2008 Ely et al; licensee BioMed Central Ltd.

A true programmable HPF hybrid three-phase rectifier

In this paper is proposed and analyzed a digital hysteresis modulation using a FPGA (Field Programmable Gate Array) device and VHDL (Hardware Description Language), applied at a hybrid three-phase rectifier with almost unitary input power factor, composed by parallel SEPIC controlled single-phase rectifiers connected to each leg of a standard 6-pulses uncontrolled diode rectifier. The digital control allows a programmable THD (Total Harmonic Distortion) at the input currents, and it makes possible that the power rating of the switching-mode converters, connected in parallel, can be a small fraction of the total average output power, in order to obtain a compact converter, reduced input current THD and almost unitary input power factor. The proposed digital control, using a FPGA device and VHDL, offers an important flexibility for the associated control technique, in order to obtain a programmable PFC (Power Factor Correction) hybrid three-phase rectifier, in agreement with the international standards (IEC, and IEEE), which impose limits for the THD of the AC (Alternate Current) line input currents. Finally, the proposed control strategy is verified through experimental results from an implemented prototype. ©2008 IEEE.

Hybrid three-phase HPF rectifier with programmable input current THD, using FPGA device and VHDL language

In this paper is proposed and analyzed a digital hysteresis modulation using a FPGA (Field Programmable Gate Array) device and VHDL (Hardware Description Language), applied at a hybrid three-phase rectifier with almost unitary input power factor, composed by parallel SEPIC controlled single-phase rectifiers connected to each leg of a standard 6-pulses uncontrolled diode rectifier. The digital control allows a programmable THD (Total Harmonic Distortion) at the input currents, and it makes possible that the power rating of the switching-mode converters, connected in parallel, can be a small fraction of the total average output power, in order to obtain a compact converter, reduced input current THD and almost unitary input power factor. Finally, the proposed digital control, using a FPGA device and VHDL, offers an important flexibility for the associated control technique, in order to obtain a programmable PFC (Power Factor Correction) hybrid three-phase rectifier, in agreement with the international standards (IEC, and IEEE), which impose limits for the THD of the AC (Alternate Current) line input currents. The proposed strategy is verified by experiments. © 2008 IEEE.

Multiloop controller and reference generator for a dynamic voltage restorer implementation

This paper deals with the design and analysis of a Dynamic Voltage Restorer output voltage control. Such control is based on a multiloop strategy, with an inner current PID regulator and an outer P+Resonant voltage controller. The inner regulator is applied on the output inductor current. It will be also demonstrated how the load current behavior may influence in the DVR output voltage, which justifies the need for the resonant controller. Additionally, it will be discussed the application of a modified algorithm for the identification of the DVR voltage references, which is based on a previously presented positive sequence detector. Since the studied three-phase DVR is assumed to be based on three identical H-bridge converters, all the analysis and design procedures were realized by means of single-phase equivalent circuits. The discussions and conclusions are supported by theoretical calculations, nonlinear simulations and some experimental results. ©2008 IEEE.

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.

Reduction of the memory size in the microprogrammed controllers

In the paper the method of reduction of the memory size in the microprogrammed controllers with sharing codes is discussed. The idea is based on the modification of internal modules and connections of the device. Next, the reduction of the microinstruction length based on the hypergraph theory is performed, thus the total size of the memory is highly reduced. © 2011 IEEE.

Virtual instrumentation: A practical approach to control and supervision process

This work describes a control and supervision application takes into account the virtual instrumentation advantages to control and supervision industrial manufacturing stations belonging to the modular production system MPS® by Festo. These stations integrate sensors, actuators, conveyor belt and other industrial elements. The focus in this approach was to replace the use of programmable logic controllers by a computer equipped with a software application based on Labview and, together, performs the functions of traditional instruments and PLCs. The manufacturing stations had their processes modeled and simulated in Petri nets. After the models were implemented in Labview environment. Tests and previous similar works in MPS® installed in Automation Laboratory, at UNESP Sorocaba campus, showed the materials and methods used in this work allow the successful use of virtual instrumentation. The results indicate the technology as an advantageous approach for the automation of industrial processes, with gains in flexibility and reduction in project cost. © 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.

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.

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)

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.