Artificial Ribonucleases: Oligonucleotides Conjugated with Metal Ion Chelates of Azacrowns

Ribonucleic acid (RNA) has many biological roles in cells: it takes part in coding, decoding, regulating and expressing of the genes as well as has the capacity to work as a catalyst in numerous biological reactions. These qualities make RNA an interesting object of various studies. Development of useful tools with which to investigate RNA is a prerequisite for more advanced research in the field. One of such tools may be the artificial ribonucleases, which are oligonucleotide conjugates that sequence-selectively cleave complementary RNA targets. This thesis is aimed at developing new efficient metal-ion-based artificial ribonucleases. On one hand, to solve the challenges related to solid-supported synthesis of metal-ion-binding conjugates of oligonucleotides, and on the other hand, to quantify their ability to cleave various oligoribonucleotide targets in a pre-designed sequence selective manner. In this study several artificial ribonucleases based on cleaving capability of metal ion chelated azacrown moiety were designed and synthesized successfully. The most efficient ribonucleases were the ones with two azacrowns close to the 3´- end of the oligonucleotide strand. Different transition metal ions were introduced into the azacrown moiety and among them, the Zn2+ ion was found to be better than Cu2+ and Ni2+ ions.

Fitoplâncton (exceto Chlorophyceae) de um lago artificial urbano no Sul do Brasil

O presente estudo tem como objetivo conhecer a composição do fitoplâncton no Lago das Tartarugas, situado no Jardim Botânico da cidade de Porto Alegre, Estado do Rio Grande do Sul. As amostragens foram realizadas mensalmente, no período de junho de 2007 a maio de 2008, em uma estação em três diferentes níveis de profundidade, na zona pelágica. Um total de 49 táxons específicos e infraespecíficos pertencentes a sete classes foram registrados. Cyanobacteria apresentou maior número de táxons (35% dos táxons identificados) seguida de Bacillariophyceae (33%) e Euglenophyceae (16,3%). São apresentadas descrições, medidas e ilustrações dos táxons, assim como a distribuição dos mesmos durante o ciclo anual.

Cianobactérias planctônicas de um lago artificial urbano na cidade de Goiânia, GO

Os criatórios de peixe do estado de Goiás são inúmeros e de intensa atividade recreativa. No entanto, estudos sobre as cianobactérias nesses ambientes são escassos, fato preocupante, uma vez que é comum notar-se intensa proliferação do fitoplâncton em pesqueiros, principalmente devido a ações antrópicas. O perigo consiste na formação de florações de espécies potencialmente tóxicas, principalmente de cianobactérias. Este trabalho visa inventariar as espécies planctônicas de cianobactérias ocorrentes em um pesqueiro (lago Jaó - um lago artificial raso) da área municipal de Goiânia (GO) (16º39'13" S-49º13'26" O). As amostragens foram realizadas nos períodos de seca (2003 a 2008) e chuva (2009), quando visualmente era evidente a ocorrência de florações. Foram aferidas variáveis climatológicas, morfométricas e limnológicas. O período de seca foi representativo nos anos amostrados apresentando no máximo 50 mm de precipitação mensal em 2005. Foram registrados 31 táxons de cianobactérias pertencentes aos gêneros Dolichospermum (5 spp.), Aphanocapsa (4 spp.), Microcystis (3 spp.), Pseudanabaena (3 spp.), Radiocystis (2 spp.), Oscillatoria (2 spp.), Bacularia, Coelosphaerium, Cylindrospermopsis, Geitlerinema, Glaucospira, Limnothrix, Pannus, Phormidium, Planktolyngbya, Planktothrix, Sphaerocavum e Synechocystis, esses últimos com uma espécie cada. Nos anos de 2003 a 2005 ocorreu predomínio de florações de espécies de Dolichospermum e em 2006 predominaram espécies de Microcystis, Radiocystis e Aphanocapsa. Das espécies inventariadas neste estudo, 21 são primeiras citações para o estado de Goiás e 13 foram constadas na literatura como potencialmente tóxicas.

Plant height reduction in populations of triticale (X triticosecale Wittmack) by induced mutations and artificial crosses

Induced mutations by gamma radiation (0, 5, 10, 20 and 40 kR doses) and reciprocal crosses were tested as mechanisms of enhancing genetic variability for plant height in two triticale cultivars, BR4 and EMBRAPA18. The reciprocal crosses and all doses of radiation showed similar increase in genetic amplitude for this trait, being suitable for increasing variability in breeding programs. Genotypes showed different responses as the gamma ray doses were increased, expressing shorter plant height. The decision of using induced mutations or artificial crosses depends on the resources available and the selection method to be used

Identification of the main generator source of longitudinal muscle contraction in the earthworm ventral nerve cord

The main generator source of a longitudinal muscle contraction was identified as an M (mechanical-stimulus-sensitive) circuit composed of a presynaptic M-1 neuron and a postsynaptic M-2 neuron in the ventral nerve cord of the earthworm, Amynthas hawayanus, by simultaneous intracellular response recording and Lucifer Yellow-CH injection with two microelectrodes. Five-peaked responses were evoked in both neurons by a mechanical, but not by an electrical, stimulus to the mechanoreceptor in the shaft of a seta at the opposite side of an epidermis-muscle-nerve-cord preparation. This response was correlated to 84% of the amplitude, 73% of the rising rate and 81% of the duration of a longitudinal muscle contraction recorded by a mechano-electrical transducer after eliminating the other possible generator sources by partitioning the epidermis-muscle piece of this preparation. The pre- and postsynaptic relationship between these two neurons was determined by alternately stimulating and recording with two microelectrodes. Images of the Lucifer Yellow-CH-filled M-1 and M-2 neurons showed that both of them are composed of bundles of longitudinal processes situated on the side of the nerve cord opposite to stimulation. The M-1 neuron has an afferent process (A1) in the first nerve at the stimulated side of this preparation and the M-2 neuron has two efferent processes (E1 and E3) in the first and third nerves at the recording side where their effector muscle cell was identified by a third microelectrode.

Facilitation of the main generator source of earthworm muscle contraction by a peripheral neuron

A constant facilitation of responses evoked in the earthworm muscle contraction generator neurons by responses evoked in the neurons of its peripheral nervous system was demonstrated. It is based on the proposal that these two responses are bifurcations of an afferent response evoked by the same peripheral mechanical stimulus but converging again on this central neuron. A single-peaked generator response without facilitation was demonstrated by sectioning the afferent route of the peripheral facilitatory modulatory response, or conditioning response (CR). The multipeaked response could be restored by restimulating the sectioned modulatory neuron with an intracellular substitutive conditioning stimulus (SCS). These multi-peaked responses were proposed to be the result of reverberating the original single peaked unconditioned response (UR) through a parallel (P) neuronal circuit which receives the facilitation of the peripheral modulatory neuron. This peripheral modulatory neuron was named "Peri-Kästchen" (PK) neuron because it has about 20 peripheral processes distributed on the surface of a Kästchen of longitudinal muscle cells on the body wall of this preparation as revealed by the Lucifer Yellow-CH-filling method.

Family ownership – a source of sustainable success in listed companies

Extant research on exchange-listed firms has acknowledged that the concentration of ownership and the identity of owners make a difference. In addition, studies indicate that firms with a dominant owner outperform firms with dispersed ownership. During the last few years, scholars have identified one group of owners, in particular, whose ownership stake in publicly listed firm is positively related to performance: the business family. While acknowledging that family firms represent a unique organizational form, scholars have identified various concepts and theories in order to understand how the family influences organizational processes and firm performance. Despite multitude of research, scholars have not been able to present clear results on how firm performance is actually impacted by the family. In other words, studies comparing the performance of listed family and other types of firms have remained descriptive in nature since they lack empirical data and confirmation from the family business representatives. What seems to be missing is a convincing theory that links the involvement and behavioral consequences. Accordingly, scholars have not yet come to a mutual understanding of what precisely constitutes a family business. The variety of different definitions and theories has made comparability of different results difficult for instance. These two issues have hampered the development of a rigorous theory of family business. The overall objective of this study is to describe and understand how the family as a dominant owner can enhance firm performance, and can act a source of sustainable success in listed companies. In more detail, in order to develop understanding of the unique factors that can act as competitive advantages for listed family firms, this study is based on a qualitative approach and aims at theory development, not theory verification. The data in this study consist of 16 thematic interviews with CEOs, members of the board, supervisory board chairs, and founders of Finnish listed-family firms. The study consists of two parts. The first part introduces the research topic, research paradigm, methods, and publications, and also discusses the overall outcomes and contributions of the publications. The second part consists of four publications that address the research questions from different viewpoints. The analyses of this study indicate that family ownership in listed companies represents a structure that differs from the traditional views of agency and stewardship, as well as from resource-based and stakeholder views. As opposed to these theories and shareholder capitalism which consider humans as individualistic, opportunistic, and self-serving, and assume that the behaviors of an investor are based on the incentives and motivations to maximize private profits, the family owners form a collective social unit that is motivated to act together toward their mutual purpose or benefit. In addition, socio-emotional and psychological elements of ownership define the family members as owners, rather than the legal and financial dimensions of ownership. That is, collective psychological ownership of family over the business (F-CPO) can be seen as a construct that comprehensively captures the fusion between the family and the business. Moreover, it captures the realized, rather than merely potential, family influence on and interaction with the business, and thereby brings more theoretical clarity of the nature of the fusion between the family and the business, and offers a solution to the problem of family business definition. This doctoral dissertation provides academics, policy-makers, family business practitioners, and the society at large with many implications considering family and business relationships.

Automated grading of left ventricular segmental wall motion by an artificial neural network using color kinesis images

The present study describes an auxiliary tool in the diagnosis of left ventricular (LV) segmental wall motion (WM) abnormalities based on color-coded echocardiographic WM images. An artificial neural network (ANN) was developed and validated for grading LV segmental WM using data from color kinesis (CK) images, a technique developed to display the timing and magnitude of global and regional WM in real time. We evaluated 21 normal subjects and 20 patients with LVWM abnormalities revealed by two-dimensional echocardiography. CK images were obtained in two sets of viewing planes. A method was developed to analyze CK images, providing quantitation of fractional area change in each of the 16 LV segments. Two experienced observers analyzed LVWM from two-dimensional images and scored them as: 1) normal, 2) mild hypokinesia, 3) moderate hypokinesia, 4) severe hypokinesia, 5) akinesia, and 6) dyskinesia. Based on expert analysis of 10 normal subjects and 10 patients, we trained a multilayer perceptron ANN using a back-propagation algorithm to provide automated grading of LVWM, and this ANN was then tested in the remaining subjects. Excellent concordance between expert and ANN analysis was shown by ROC curve analysis, with measured area under the curve of 0.975. An excellent correlation was also obtained for global LV segmental WM index by expert and ANN analysis (R² = 0.99). In conclusion, ANN showed high accuracy for automated semi-quantitative grading of WM based on CK images. This technique can be an important aid, improving diagnostic accuracy and reducing inter-observer variability in scoring segmental LVWM.

Allogeneic hematopoietic stem cell transplantation from an alternative stem cell source in Fanconi anemia patients: analysis of 47 patients from a single institution

We transplanted 47 patients with Fanconi anemia using an alternative source of hematopoietic cells. The patients were assigned to the following groups: group 1, unrelated bone marrow (N = 15); group 2, unrelated cord blood (N = 17), and group 3, related non-sibling bone marrow (N = 15). Twenty-four patients (51%) had complete engraftment, which was not influenced by gender (P = 0.87), age (P = 0.45), dose of cyclophosphamide (P = 0.80), nucleated cell dose infused (P = 0.60), or use of anti-T serotherapy (P = 0.20). Favorable factors for superior engraftment were full HLA compatibility (independent of the source of cells; P = 0.007) and use of a fludarabine-based conditioning regimen (P = 0.046). Unfavorable factors were > or = 25 transfusions pre-transplant (P = 0.011) and degree of HLA disparity (P = 0.007). Intensity of mucositis (P = 0.50) and use of androgen prior to transplant had no influence on survival (P = 0.80). Acute graft-versus-host disease (GVHD) grade II-IV and chronic GVHD were diagnosed in 47 and 23% of available patients, respectively, and infections prevailed as the main cause of death, associated or not with GVHD. Eighteen patients are alive, the Kaplan-Meyer overall survival is 38% at ~8 years, and the best results were obtained with related non-sibling bone marrow patients. Three recommendations emerged from the present study: fludarabine as part of conditioning, transplant in patients with <25 transfusions and avoidance of HLA disparity. In addition, an extended family search (even when consanguinity is not present) seeking for a related non-sibling donor is highly recommended.

Artificial reflecting structures based on metamaterials

This thesis studies metamaterial-inspired mirrors which provide the most general control over the amplitude and phase of the reflected wavefront. The goal is to explore practical possibilities in designing fully reflective electromagnetic structures with full control over reflection phase. The first part of the thesis describes a planar focusing metamirror with the focal distance less than the operating wavelength. Its practical applicability from the viewpoint of aberrations when the incident angle deviates from the normal one is verified numerically and experimentally. The results indicate that the proposed focusing metamirror can be efficiently employed in many different applications due to its advantages over other conventional mirrors. In the second part of the thesis a new theoretical concept of reflecting metasurface operation is introduced based on Huygens’ principle. This concept in contrast to known approaches takes into account all the requirements of perfect metamirror operation. The theory shows a route to improve the previously proposed metamirrors through tilting the individual inclusions of the structure at a chosen angle from normal. It is numerically tested and the results demonstrate improvements over the previous design.

The husk fiber of Cocos nucifera L. (Palmae) is a source of anti-neoplastic activity

In the present study, we investigated the in vitro anti-tumoral activities of fractions from aqueous extracts of the husk fiber of the typical A and common varieties of Cocos nucifera (Palmae). Cytotoxicity against leukemia cells was determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Cells (2 x 104/well) were incubated with 0, 5, 50 or 500 µg/mL high- or low-molecular weight fractions for 48 h, treated with MTT and absorbance was measured with an ELISA reader. The results showed that both varieties have almost similar antitumoral activity against the leukemia cell line K562 (60.1 ± 8.5 and 47.5 ± 11.9% for the typical A and common varieties, respectively). Separation of the crude extracts with Amicon membranes yielded fractions with molecular weights ranging in size from 1-3 kDa (fraction A) to 3-10 kDa (fraction B) and to more than 10 kDa (fraction C). Cells were treated with 500 µg/mL of these fractions and cytotoxicity was evaluated by MTT. Fractions ranging in molecular weight from 1-10 kDa had higher cytotoxicity. Interestingly, C. nucifera extracts were also active against Lucena 1, a multidrug-resistant leukemia cell line. Their cytotoxicity against this cell line was about 50% (51.9 ± 3.2 and 56.3 ± 2.9 for varieties typical A and common, respectively). Since the common C. nucifera variety is extensively cultured in Brazil and the husk fiber is its industrial by-product, the results obtained in the present study suggest that it might be a very inexpensive source of new antineoplastic and anti-multidrug resistant drugs that warrants further investigation.

Synergistic control of forearm based on accelerometer data and artificial neural networks

In the present study, we modeled a reaching task as a two-link mechanism. The upper arm and forearm motion trajectories during vertical arm movements were estimated from the measured angular accelerations with dual-axis accelerometers. A data set of reaching synergies from able-bodied individuals was used to train a radial basis function artificial neural network with upper arm/forearm tangential angular accelerations. The trained radial basis function artificial neural network for the specific movements predicted forearm motion from new upper arm trajectories with high correlation (mean, 0.9149-0.941). For all other movements, prediction was low (range, 0.0316-0.8302). Results suggest that the proposed algorithm is successful in generalization over similar motions and subjects. Such networks may be used as a high-level controller that could predict forearm kinematics from voluntary movements of the upper arm. This methodology is suitable for restoring the upper limb functions of individuals with motor disabilities of the forearm, but not of the upper arm. The developed control paradigm is applicable to upper-limb orthotic systems employing functional electrical stimulation. The proposed approach is of great significance particularly for humans with spinal cord injuries in a free-living environment. The implication of a measurement system with dual-axis accelerometers, developed for this study, is further seen in the evaluation of movement during the course of rehabilitation. For this purpose, training-related changes in synergies apparent from movement kinematics during rehabilitation would characterize the extent and the course of recovery. As such, a simple system using this methodology is of particular importance for stroke patients. The results underlie the important issue of upper-limb coordination.

EEG spike source localization before and after surgery for temporal lobe epilepsy: a BOLD EEG-fMRI and independent component analysis study

Simultaneous measurements of EEG-functional magnetic resonance imaging (fMRI) combine the high temporal resolution of EEG with the distinctive spatial resolution of fMRI. The purpose of this EEG-fMRI study was to search for hemodynamic responses (blood oxygen level-dependent - BOLD responses) associated with interictal activity in a case of right mesial temporal lobe epilepsy before and after a successful selective amygdalohippocampectomy. Therefore, the study found the epileptogenic source by this noninvasive imaging technique and compared the results after removing the atrophied hippocampus. Additionally, the present study investigated the effectiveness of two different ways of localizing epileptiform spike sources, i.e., BOLD contrast and independent component analysis dipole model, by comparing their respective outcomes to the resected epileptogenic region. Our findings suggested a right hippocampus induction of the large interictal activity in the left hemisphere. Although almost a quarter of the dipoles were found near the right hippocampus region, dipole modeling resulted in a widespread distribution, making EEG analysis too weak to precisely determine by itself the source localization even by a sophisticated method of analysis such as independent component analysis. On the other hand, the combined EEG-fMRI technique made it possible to highlight the epileptogenic foci quite efficiently.

Artificial neural network models for predicting 1-year mortality in elderly patients with intertrochanteric fractures in China

The mortality rate of older patients with intertrochanteric fractures has been increasing with the aging of populations in China. The purpose of this study was: 1) to develop an artificial neural network (ANN) using clinical information to predict the 1-year mortality of elderly patients with intertrochanteric fractures, and 2) to compare the ANN's predictive ability with that of logistic regression models. The ANN model was tested against actual outcomes of an intertrochanteric femoral fracture database in China. The ANN model was generated with eight clinical inputs and a single output. ANN's performance was compared with a logistic regression model created with the same inputs in terms of accuracy, sensitivity, specificity, and discriminability. The study population was composed of 2150 patients (679 males and 1471 females): 1432 in the training group and 718 new patients in the testing group. The ANN model that had eight neurons in the hidden layer had the highest accuracies among the four ANN models: 92.46 and 85.79% in both training and testing datasets, respectively. The areas under the receiver operating characteristic curves of the automatically selected ANN model for both datasets were 0.901 (95%CI=0.814-0.988) and 0.869 (95%CI=0.748-0.990), higher than the 0.745 (95%CI=0.612-0.879) and 0.728 (95%CI=0.595-0.862) of the logistic regression model. The ANN model can be used for predicting 1-year mortality in elderly patients with intertrochanteric fractures. It outperformed a logistic regression on multiple performance measures when given the same variables.

HYBRID ARTIFICIAL NEURAL NETWORK APPLIEDTO MODELING SCFE OF BASIL AND ROSEMARY OILS

This work presents the results of a Hybrid Neural Network (HNN) technique as applied to modeling SCFE curves obtained from two Brazilian vegetable matrices. A series Hybrid Neural Network was employed to estimate the parameters of the phenomenological model. A small set of SCFE data of each vegetable was used to generate an extended data set, sufficient to train the network. Afterwards, other sets of experimental data, not used in the network training, were used to validate the present approach. The series HNN correlates well the experimental data and it is shown that the predictions accomplished with this technique may be promising for SCFE purposes.