Development of triplet repeat primed PCR (TP-PCR) technique as a support of molecular test in Machado-Joseph disease

Dissertação de Mestrado, Ciências Biomédicas, 3 de Fevereiro de 2016, Universidade dos Açores.

The influence of the blood pressure on the venous cerebral flow measured by magnetic susceptibility (SWI) technique

Susceptibility Weighted Image (SWI) is a Magnetic Resonance Imaging (MRI) technique that combines high spatial resolution and sensitivity to provide magnetic susceptibility differences between tissues. It is extremely sensitive to venous blood due to its iron content of deoxyhemoglobin. The aim of this study was to evaluate, through the SWI technique, the differences in cerebral venous vasculature according to the variation of blood pressure values. 20 subjects divided in two groups (10 hypertensive and 10 normotensive patients) underwent a MRI system with a Siemens® scanner model Avanto of 1.5T using a synergy head coil (4 channels). The obtained sequences were T1w, T2w-FLAIR, T2* and SWI. The value of Contrast-to-Noise Ratio (CNR) was assessed in MinIP (Minimum Intensity Projection) and Magnitude images, through drawing free hand ROIs in venous structures: Superior Sagittal Sinus (SSS) Internal Cerebral Vein (ICV) and Sinus Confluence (SC). The obtained values were presented in descriptive statistics-quartiles and extremes diagrams. The results were compared between groups. CNR shown higher values for normotensive group in MinIP (108.89 ± 6.907) to ICV; (238.73 ± 18.556) to SC and (239.384 ± 52.303) to SSS. These values are bigger than images from Hypertensive group about 46 a.u. in average. Comparing the results of Magnitude and MinIP images, there were obtained lower CNR values for the hypertensive group. There were differences in the CNR values between both groups, being these values more expressive in the large vessels-SSS and SC. The SWI is a potential technique to evaluate and characterize the blood pressure variation in the studied vessels adding a physiological perspective to MRI and giving a new approach to the radiological vascular studies.

Does diaphragmatic breathing technique really improve its range of motion? An objective assessment

Purpose - To verify the results of a diaphragmatic breathing technique (DBT) on diaphragmatic range of motion in healthy subjects. Methods - A total of 51 healthy subjects (10 male; 41 female), mean age 20 years old and a body mass index (BMI) ranging from 15.6 to 34.9 kg/m2, were enrolled in this study. Diaphragmatic range of motion was assessed by M-mode ultrasound imaging. Measurements were made before and after the DBT implementation in a standard protocol, based on 3 seconds of inspiration starting from a maximum expiration. Differences between assessments were analyzed by descriptive statistics and t-test (p < 0.05). Results - Mean value range of motion before DBT was 55.3 ± 13.4 mm and after DBT was 63.8 ± 13.2 mm showing a significant improvement of 8.5 ± 14.7 mm (p < 0.001). A strong correlation between the slope and the range of motion was found (r = 0.71, p < 0.001). Conclusions - Based on ultrasound measurements, it has been proved that DBT really contributes to a higher diaphragmatic range of motion. Future studies are needed in order to understand the influence of protocol parameters (e.g. inspiration time). Clinical implications - In the contest of evidence-based practice in physiotherapy, it has been showed by objective measurements that DBT improves the diaphragm range of motion, translating into a more efficient ventilatory function and thus can be used in clinical setting. To our knowledge this is the first study to assess the effects of DBT on range of motion of diaphragm muscle with ultrasound imaging.

Solid-State bipolar Marx modulator modeling

A mathematical model that simulates the operation of a solid-state bipolar Marx modulator topology, including the influence of parasitic capacitances is presented and discussed as a tool to analyze the circuit behavior and to assist the design engineer to select the semiconductor components and to enhance the operating performance. Simulations show good agreement with experimental results, considering a four stage circuit assembled with 1200 V isolated gate bipolar transistors and diodes, operating at 1000 V dc input voltage and 1-kHz frequency, giving 4 kV and 10-mu s output pulses into several resistive loads. Results show that parasitic capacitances between Marx cells to ground can significantly load the solid-state switches, adding new operating circuit conditions.

A sensor classification strategy for robotic manipulators using multidimensional scaling technique

This paper analyzes the signals captured during impacts and vibrations of a mechanical manipulator. To test the impacts, a flexible beam is clamped to the end-effector of a manipulator that is programmed in a way such that the rod moves against a rigid surface. Eighteen signals are captured and theirs correlation are calculated. A sensor classification scheme based on the multidimensional scaling technique is presented.

Digital-domain self-calibration technique for video-rate pipeline A/D converters using Gaussian white noise

Electronics Letters Vol.38, nº 19

Comparison between monopolar and bipolar microsecond range pulsed electric fields in enhancement of apple juice extraction

The effect of monopolar and bipolar shaped pulses in additional yield of apple juice extraction is evaluated. The applied electric field strength, pulsewidth, and number of pulses are assessed for both pulse types, and divergences are analyzed. Variation of electric field strength is ranged from 100 to 1300 V/cm, pulsewidth from 20 to 300 mu s, and the number of pulses from 10 to 200, at a frequency of 200 Hz. Two pulse trains separated by 1 s are applied to apple cubes. Results are plotted against reference untreated samples for all assays. Specific energy consumption is calculated for each experiment as well as qualitative indicators for apple juice of total soluble dry matter and absorbance at 390-nm wavelength. Bipolar pulses demonstrated higher efficiency, and specific energetic consumption has a threshold where higher inputs of energy do not result in higher juice extraction when electric field variation is applied. Total soluble dry matter and absorbance results do not illustrate significant differences between application of monopolar and bipolar pulses, but all values are inside the limits proposed for apple juice intended for human consumption.

Marx-Type solid-state bipolar modulator topologies: Performance comparison

The operation of generalized Marx-type solid-state bipolar modulators is discussed and compared with simplified Marx-derived circuits, to evaluate their capability to deal with various load conditions. A comparative analysis on the number of switches per cell, fiber optic trigger count, losses, and switch hold-off voltages has been made. A circuit topology is obtained as a compromise in terms of operating performance, trigger simplicity, and switching losses. A five-stage laboratory prototype of this circuit has been assembled using 1200 V insulated gate bipolar transistors (IGBTs) and diodes, operating with 1000 V dc input voltage and 1 kHz frequency, giving 5 kV bipolar pulses, with 2.5 mu s pulse width and 5 mu s relaxation time into resistive, capacitive, and inductive loads.

Modeling of a solid-state bipolar blumlein generator for N stages

This paper models an n-stage stacked Blumlein generator for bipolar pulses for various load conditions. Calculation of the voltage amplitudes in time domain at the load and between stages is described for an n-stage generator. For this, the reflection and transmission coefficients are mathematically modeled where impedance discontinuity occurs (i.e., at the junctions between two transmission lines). The mathematical model developed is assessed by comparing simulation results to experimental data from a two-stage Blumlein solid-state prototype.

Pulsed electric field pre-treatment for apple juice extraction: Evaluation of monopolar and bipolar pulses effects

Additional apple juice extraction with pulsed electric field pretreated apple cubes towards control samples is evaluated. Monopolar and bipolar shaped pulses are compared and their effect is studied with variation of electric field, pulse width and number of pulses. Variation of electric field strength is ranged from 100 V/cm to 1300 V/cm, pulse width from 20 mu s to 300 mu s and number of pulses from 10 to 200, at frequency of 200Hz. Two pulse trains separated by 1 second are applied to all samples. Bipolar pulses showed higher apple juice yields with all studied parameters. Calculation of specific energies consumed was assessed and a threshold where higher energy inputs do not increase juice yield is found for a number of used parameters. Qualitative parameters of total soluble matter (Brix) and absorbance at 390 nm wavelength were determined for each sample and results show that no substantial differences are found for PEF pre-treated and control samples.

Immunoperoxidase technique in experimental chronic chagasic myocarditis

Chagas'disease has been described as the commonest form of chronic myocarditis. An immunologic pathogenesis has been discribed for this form of the disease. So far, no immunoperoxidase technique has been used for the detection of immunological deposits in chronic experimental Chagas'myocardiopathy. Forty-one Swiss mice, three months old were inoculated intraperitoneally with doses between 10 and 10(5) Tulahuen trypomastigotes. Mice were reinoculated one month after with doses between 10² and 10(5) and sacrificed at 6 (n=21) and 9 months (n=9) after the first inoculation. ECGs were recorded before sacrifice. Immunoperoxidase technique (peroxidase-antiperoxidase method), immunofluorescence (direct and indirect) as well as histological studies were performed in myocardiums and skeletal muscles of the surviving animals. The most sensitive methods for detecting chronic chagasic infection were the routine histologic studies (73%) and the ECGs 83% and 89% on 6 and 9 mo. post-infected mice, respectively. Myocardial involvement varied from interstitial mild focal lymphocyte infiltrates up to replacement of myocytes by loose connective tissue. Atrial myocardiums (21/23, 91%) were more affected than ventricles (9/23, 39%). Typical chagasic nests were rarely found. Skeletal muscle involvement (11/18 and 7/9) varied from mild to extensive lymphocyte and plasmacell infiltrates, and necrotic fibers. The involved antigen were shown in skeletal muscles by the immunoperoxidase technique as diffusely arranged granular intracytoplasmatic deposit for both IgC and total immunoglobulins. The coincidence between this technique and histologic muscle lesions was 11/18 (61(%) in 6 mo. and 6/8 (75%) at 9 mo. post-infection. In heart, delicate granular deposits of total immunoglobulins were seen diffusely arranged within the ventricular myocytes; coincidence between immunoperoxidase technique anl histologic involvement increased from 36 to 66% in animals sacrifeced 6 and 9 mo. post-infection. This strongly stressed the increase of immunologic phenomena with the chronification of infection. Concerning sensitivity, immunoperoxidase and direct immunofluorescence were highly sensitive in skeletal muscle (100%, p < 0.01). Conversely, direct immunofluorescence technique showed poor results in heart while immunoperoxidase increased its sensitivity from 21.4% (at 6 mo.) to 66.6% (at 9 mo.) post-infection (p < 0.001). Considering the necessity of obtaining an adequate vaccine in order to prevent this disease an experimental model like this, rendering immunological reactions as revealed by the immunoperoxidase technique, would be useful.

Therapeutical evaluation of different dose regimens of praziquantel in schistosomiasis mansoni, based on the quantitative oogram technique

A clinical trial involving 80 patients of both sexes, from ages 15 to 55, with chronic intestinal or hepatointestinal schistosomiasis mansoni, was carried out to evaluate the therapeutical efficacy of different dose regimens of praziquantel. The patients were randomly allocated into four groups with an equal number of cases and were then treated with one of the following dosages: 60 mg/kg for 1 day; 60 mg/kg daily for 2 days; 60 mg/kg daily for 3 days; and 30 mg/kg daily for 6 days. The assessment of parasitological cure was based on the quantitative oogram technique through rectal mucosa biopsies which were undertaken prior to, as well as, 1,2,4 and 6 months post-treatment. Concurrently, stool examinations according to the qualitative Hoffman, Pons & Janer (HPJ) and the quantitative Kato-Katz (K-K) methods were also performed. The best tolerability was observed with 30 mg/kg daily for 6 days whereas the highest incidence of side-effects (mainly dizziness and nausea) was found with 60 mg/kg daily for 3 days. No serious adverse drug reaction has occurred. The achieved cure rates were: 25% with 60 mg/kg for 1 day; 60% with 60 mg/kg daily for 2 days; 89.5% with 60 mg/kg daily for 3 days; and 90% with 30 mg/kg daily for 6 days. At the same time there has been a downfall of 64%, 73%, 87% and 84% respectively, in the median number of viable S. mansoni ova per gram of tissue. Thus, a very clear direct correlation between dose and effect could be seen. The corresponding cure rates according to stool examinations by HPJ were 39%, 80%, 100% and 95%; by K-K 89%, 100%, 100% and 100%. This discrepancy in results amongst the three parasitological methods is certainly due to their unequal accuracy. In fact, when the number of viable eggs per gram of tissue fell below 5,000 the difference in the percentage of false negative findings between HPJ (28%) and K-K (80%) became significative. When this number dropped to less than 2,000 the percentage of false negative results obtained with HPJ (49%) turned significant in relation to the oogram as well. In conclusion, it has been proven that praziquantel is a highly efficacious agent against S. mansoni infections. If administered at a total dose of 180 mg/kg divided into either 3 or 6 days, it yields a 90% cure rate. Possibly, one could reach 100% by increasing the total dose to 240 mg/kg. Furthermore, it was confirmed that the quantitative oogram technique is the most reliable parasitological method when evaluating the efficacy of new drugs in schistosomiasis mansoni.

Construção de um variador de frequência

A dificuldade de controlo de um motor de indução, bem como o armazenamento de energia CC e posterior utilização como energia alternada promoveram o desenvolvimento de variadores de frequência e inversores. Assim, como projeto de tese de mestrado em Automação e Sistemas surge o desenvolvimento de um variador de frequência. Para elaboração do variador de frequência efetuou-se um estudo sobre as técnicas de modulação utilizadas nos inversores. A técnica escolhida e utilizada é a Sinusoidal Pulse Width Modulation (SPWM). Esta técnica baseia-se na modelação por largura de impulso (PWM), o qual é formado por comparação de um sinal de referência com um sinal de portadora de elevada frequência. Por sua vez, a topologia escolhida para o inversor corresponde a um Voltage Source Inverter (VSI) de ponte trifásica completa a três terminais. O desenvolvimento da técnica de modulação SPWM levou ao desenvolvimento de um modelo de simulação em SIMULINK, o qual permitiu retirar conclusões sobre os resultados obtidos. Na fase de implementação, foram desenvolvidas placas para o funcionamento do variador de frequência. Assim, numa fase inicial foi desenvolvida a placa de controlo, a qual contém a unidade de processamento e que é responsável pela atuação de Insulated Gate Bipolar Transistors (IGBTs). Para além disso, foi desenvolvida uma placa para proteção dos IGBTs (evitando condução simultânea no mesmo terminal) e uma placa de fontes isoladas para alimentação dos circuitos e para atuação dos IGBTs. Ainda, foi desenvolvida a técnica de SPWM em software para a unidade de controlo e finalmente foi desenvolvida uma interface gráfica para interação com o utilizador. A validação do projeto foi conseguida através da variação da velocidade do motor de indução trifásico. Para isso, este foi colocado a funcionar a diversas frequências de funcionamento e a diferentes amplitudes. Para além disso, o seu funcionamento foi também validado utilizando uma carga trifásica equilibrada de 3 lâmpadas de forma a ser visualizada a variação de frequência e variação de amplitude.

Identification of mating type genes in the bipolar basidiomycetous yeast rhodosporidium toruloides: first insight into the MAT Locus structure of the sporidiobolales

Eukaryotic Cell, Vol.7, Nº6