Development of new methodologies in sample treatment for proteomics workflow based on enzymatic probe sonication technology and mass spectrometry

Thesis submitted to the Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia, for the degree of Doctor of Philosophy in Biochemistry

Evaluation of ultrasound, ultrasonophoresis and homeopathic mesotherapy on Cellulite treatment

Introduction: Cellulite is a complex architectural disorder with multifactorial etiologies that is prevalent in 98% of the women (1). Nowadays several aesthetic treatments are being used: surgical, cosmetic, physical, mechanical, and thermal. (2) Most treatments lack a substantial proof of efficacy. Objective: The purpose of this study was to test and evaluate the efficacy of Ultrasound, Homeopathic Ultrasonophoresis, and Homeopathic Mesotherapy versus control in cellulite in a population of women from ESTSP. Methods: Female volunteers (n=23), Caucasian, aged between 18-31 years, with BMI 19-27 kg/m2 with clinical cellulite gradation on the Cellulite Grading Scale of 1 to 4 were included in a control controlled study. Subjects were assigned in four different groups: Group I (Control, n=6), Group II (Ultrasound, (n=5), Group III (Homeopathic Ultrasonophoresis, n=6), Group IV (Homeopathic Mesotherapy, n=6). Groups II to IV were treated 3 times per week, for a total of 10 sessions. Cellulite gradation was evaluated at the beginning and the end of the trial by means of clinical photography, using a Canon IXUS 65 (6 mega pixels). For homeopathic treatments Dr. Reckeweg® Rekin® 59, 13 and 42 – Dietmed were used. The rating of perceived pain during Homeopathic Mesotherapy was evaluated by a visual analogic scale (VAS). The equipment Sonopuls 692, Enraf-Nonius was used for Ultrasound and Ultrasonophoresis treatments. Results:The higher number of participants with improvement in cellulite graduation occurred in group II (80%), followed group III (50%) and by group IV (33%). The group in which more changes in cellulite gradation occurred was group II, 20% of the individuals improved their score in 2 points. Results were statistically different between Group I and Group II, p=0,015. During the treatments of homeopathic mesotherapy the pain diminished 1 value in VAS scale. Discussion and Conclusion: Although all the three interventions groups were effective in the improvement of cellulite, as expected from previous works described in the literature, (2) only the ultrasound group was statistically different from control. These preliminary results point to the need of a new study using a higher number of participants and the same methodology.

Synthesis about a collaborative project on “Technology Assessment of Autonomous System

The project started in 2009 with the support of DAAD in Germany and CRUP in Portugal under the “Collaborative German-Portuguese University Actions” programme. One central goal is the further development of a theory of technology assessment applied to robotics and autonomous systems in general that reflects in its methodology the changing conditions of knowledge production in modern societies and the emergence of new robotic technologies and of associated disruptive changes. Relevant topics here are handling broadened future horizons and new clusters of science and technology (medicine, engineering, interfaces, industrial automation, micro-devices, security and safety), as well as new governance structures in policy decision making concerning research and development (R

Development of molecularly imprinted polymers using supercritical fluid technology

Dissertação para obtenção do Grau de Doutor em Química Sustentável

Synthesis of molecular biomimetics

Biomimetics has paved the way toward new materials and technologies inspired in Nature. Biomolecules and their supramolecular organization have today a leading role in biomimetics, benefiting from the recent advances in nanotechnology. The production of biomimetic materials may be however a difficult task, because Nature does it very well. The use of several building blocks assembled in bottom-up arrangement is without doubt at the core of this process. Such building blocks include different molecules or molecular arrangements, of synthetic or natural origin, such as amino acids, lipids, carbohydrates, nucleic acids, carbon allotropes, dendrimers, or organosilanes, among others. The most common approaches to produce synthetic biomimetic materials are reported herein, with special emphasis to building blocks and their supramolecular arrangement.

Synthesis of an antibody-like material for the detection of Albumin

6th Graduate Student Symposium on Molecular Imprinting

Cyclopentadienyl-functionalised N-heterocyclic carbenes: synthesis, coordination to Mo, Ru, Rh

Dissertation presented to obtain the Ph.D degree in Chemistry

Automated Synthesis Procedure of RF Discrete Tuning Differential Capacitance Circuits

The paper presents a RFDSCA automated synthesis procedure. This algorithm determines several RFDSCA circuits from the top-level system specifications all with the same maximum performance. The genetic synthesis tool optimizes a fitness function proportional to the RFDSCA quality factor and uses the epsiv-concept and maximin sorting scheme to achieve a set of solutions well distributed along a non-dominated front. To confirm the results of the algorithm, three RFDSCAs were simulated in SpectreRF and one of them was implemented and tested. The design used a 0.25 mum BiCMOS process. All the results (synthesized, simulated and measured) are very close, which indicate that the genetic synthesis method is a very useful tool to design optimum performance RFDSCAs.

New Concepts Towards the Synthesis of Digital Circuits Through Genetic Algorithms

This paper analyses the performance of a Genetic Algorithm using two new concepts, namely a static fitness function including a discontinuity measure and a fractional-order dynamic fitness function, for the synthesis of combinational logic circuits. In both cases, experiments reveal superior results in terms of speed and convergence to achieve a solution.

Vision-Based Assisted Teleoperation for Inspection Tasks with a Small ROV

It is well-known that ROVs require human intervention to guarantee the success of their assignment, as well as the equipment safety. However, as its teleoperation is quite complex to perform, there is a need for assisted teleoperation. This study aims to take on this challenge by developing vision-based assisted teleoperation maneuvers, since a standard camera is present in any ROV. The proposed approach is a visual servoing solution, that allows the user to select between several standard image processing methods and is applied to a 3-DOF ROV. The most interesting characteristic of the presented system is the exclusive use of the camera data to improve the teleoperation of an underactuated ROV. It is demonstrated through the comparison and evaluation of standard implementations of different vision methods and the execution of simple maneuvers to acquire experimental results, that the teleoperation of a small ROV can be drastically improved without the need to install additional sensors.

Bioelectrochemically-assisted reductive dechlorination of 1,2-dichloroethane by a Dehalococcoides-enriched microbial culture

The aim of this study was to verify the possibility to use a polarized graphite electrode as an electron donor for the reductive dechlorination of 1,2-dichloroethane, an ubiquitous groundwater contaminant. The rate of 1,2-DCA dechlorination almost linearly increased by decreasing the set cathode potential over a broad range of set cathode potentials (i.e., from −300 mV to −900 mV vs. the standard hydrogen electrode). This process was primarily dependent on electrolytic H2 generation. On the other hand, reductive dechlorination proceeded (although quite slowly) with a very high Coulombic efficiency (near 70%) at a set cathode potential of −300 mV, where no H2 production occurred. Under this condition, reductive dechlorination was likely driven by direct electron uptake from the surface of the polarized electrode. Taken as a whole, this study further extends the range of chlorinated contaminants which can be treated with bioelectrochemical systems.

Abdominal ultrasound in the evaluation of fibrosis and portal hypertension in an area of schistosomiasis low endemicity

This study was undertaken in the municipality of Bananal, São Paulo, an endemic area for schistosomiasis with a prevalence under 10% and low parasite load among infected individuals. Our objective was to identify the clinical forms of schistosomiasis among 109 patients in whom the disease had been diagnosed through direct fecal analysis and who had been medicated with oxamniquine at the time of the Plan for the Intensification of Schistosomiasis Control Actions (1998-2000). These patients were submitted to an abdominal ultrasonography and fecal analysis by Kato-Katz method, four years, on average, after the end of the Plan. Five patients, whose abdominal ultrasound images were compatible with either peripheral or central periportal fibrosis and portal hypertension, were identified. None of the 109 patients presented Schistosoma mansoni eggs at fecal analysis. Ultrasonography is a sensitive, noninvasive diagnostic method that allows a better identification of the extent of liver involvement in schistosomiasis cases.

Minimal Criteria for the Diagnosis of Avulsion of the Puborectalis Muscle by Tomographic Ultrasound

Introduction and hypothesis Puborectalis avulsion is a likely etiological factor for female pelvic organ prolapse(FPOP). We performed a study to establish minimal sonographic criteria for the diagnosis of avulsion. Methods We analysed datasets of 764 women seen at a urogynecological service. Offline analysis of ultrasound datasets was performed blinded to patient data. Tomographic ultrasound imaging (TUI) was used to diagnose avulsion of the puborectalis muscle. Results Logistic regression modelling of TUI data showed that complete avulsion is best diagnosed by requiring the three central tomographic slices to be abnormal. This finding was obtained in 30% of patients and was associated with symptoms and signs of FPOP (P<0.001). Lesser degrees of trauma (‘partial avulsion’) were not associated with symptoms or signs of pelvic floor dysfunction. Conclusions Complete avulsion of the puborectalis muscle is best diagnosed on TUI by requiring all three central slices to be abnormal. Partial trauma seems of limited clinical relevance.

Biosynthesis-Assisted Structural Elucidation of the Bartolosides, Chlorinated Aromatic Glycolipids from Cyanobacteria

The isolation of the bartolosides, unprecedented cyanobacterial glycolipids featuring aliphatic chains with chlorine substituents and C-glycosyl moieties, is reported. Their chlorinated dialkylresorcinol (DAR) core presented a major structural-elucidation challenge. To overcome this, we discovered the bartoloside (brt) biosynthetic gene cluster and linked it to the natural products through in vitro characterization of the DAR-forming ketosynthase and aromatase. Bioinformatic analysis also revealed a novel potential halogenase. Knowledge of the bartoloside biosynthesis constrained the DAR core structure by defining key pathway intermediates, ultimately allowing us to determine the full structures of the bartolosides. This work illustrates the power of genomics to enable the use of biosynthetic information for structure elucidation.

Laparoscopic Assisted Vaginal Hysterectomy Versus Vaginal Hysterectomy

Introduction: Hysterectomy is the commonest gynecologic operation, performed for malignant and benign conditions. There are many approaches to hysterectomy for benign disease. Studies comparing the techniques have showed that vaginal hysterectomy has benefits in terms of reduced hospital stay, faster recovery and less operating time. Objective: The purpose of this study is to compare the surgical and immediate postoperative outcomes of Laparoscopic Assisted Vaginal Hysterectomy (LAVH) with those of Vaginal Hysterectomy (VH). Methods: Retrospective descriptive study, comparing two groups of women who underwent LAVH or VH in our department during a 24 months period, from January 2009 to December 2010. The two groups were compared regarding age, vaginal deliveries, previous abdominal surgery, uterine and adnexal pathology, intra-operative and post-operative complications, uterus weight, blood loss and number of days until discharge. Results: In our study 42 LAVH and 99 VH were included, with a patient mean age of 47 and 59, respectively. The most frequent indication for hysterectomy was fibroids (80%) for LAVH and POP(58.6%) for HV. In LAVH group 47.6% of patients had previous abdominal surgery, vs 28.2% in VH group. The medium operative time was 167 minutes for LAVH vs 99 minutes for HV. The intra-operative complications were one case (2%) of accidental incision of rectum in LAVH, and one bladder incision in the VH (1%). There were 3 conversions to laparotomy for difficult technique (7%) in LAVH group. There were no significant post-operative complications for LAVH. In VH group there were 2 cases of haemoperitoneum (2%) and 1 case requiring blood transfusion (1%). The mean time for discharge was 4.23 days for LAVH and 4.46 days for VH. Conclusions: In our study, the main advantage for VH was the reduced operative time. In terms of time to discharge there was no difference between the 2 groups. The main intra-operative complication of LAVH was the risk of conversion to laparotomy, but post-operatively this procedure had fewer complications than VH. In conclusion, LAVH is a safe option for women requiring hysterectomy in cases where VH is anticipated to be technically difficult.