Five egg-laying queens in a single colony of brazilian stingless bees (Melipona scutellaris Latreille

Polygyny, characterized by the presence of several egg-laying queens, is considered as a temporary colony status. In stingless bees it is rarely observed. This paper reports the first case of natural polygyny in Melipona scutellaris colony, with five egg-laying queens.

Forecasting human entrances at a commercial store using facial recognition data

Dissertação de mestrado integrado em Engenharia e Gestão de Sistemas de Informação

Hand gesture recognition system based in computer vision and machine learning: Applications on human-machine interaction

Tese de Doutoramento em Engenharia de Eletrónica e de Computadores

Electronic tongue: a versatile tool for mineral and fruit-flavored waters recognition

Natural mineral waters (still), effervescent natural mineral waters (sparkling) and aromatized waters with fruit-flavors (still or sparkling) are an emerging market. In this work, the capability of a potentiometric electronic tongue, comprised with lipid polymeric membranes, to quantitatively estimate routinely quality physicochemical parameters (pH and conductivity) as well as to qualitatively classify water samples according to the type of water was evaluated. The study showed that a linear discriminant model, based on 21 sensors selected by the simulated annealing algorithm, could correctly classify 100 % of the water samples (leave-one out cross-validation). This potential was further demonstrated by applying a repeated K-fold cross-validation (guaranteeing that at least 15 % of independent samples were only used for internal-validation) for which 96 % of correct classifications were attained. The satisfactory recognition performance of the E-tongue could be attributed to the pH, conductivity, sugars and organic acids contents of the studied waters, which turned out in significant differences of sweetness perception indexes and total acid flavor. Moreover, the E-tongue combined with multivariate linear regression models, based on sub-sets of sensors selected by the simulated annealing algorithm, could accurately estimate waters pH (25 sensors: R 2 equal to 0.99 and 0.97 for leave-one-out or repeated K-folds cross-validation) and conductivity (23 sensors: R 2 equal to 0.997 and 0.99 for leave-one-out or repeated K-folds cross-validation). So, the overall satisfactory results achieved, allow envisaging a potential future application of electronic tongue devices for bottled water analysis and classification.

Importancia del Ca2+ en la fisiología espermática: modulación de su actividad por progesterona y proteínas del plasma seminal. Importance of calcium in sperm physiology: its regulation by progesterone and proteins from the seminal plasma.

El objetivo general del proyecto es estudiar el efecto de la progesterona y de algunas proteínas del plasma seminal sobre la actividad del Ca2+ en diferentes procesos fisiológicos que ocurren en el espermatozoide, los cuales están estrechamente relacionados con la capacidad fertilizante de esta célula. La progesterona, principal esteroide secretado por las células del cumulus oophorus, ejerce su efecto a través de un receptor no-genómico provocando aumento en el calcio intracelular de los espermatozoides y, consecuentemente, promoviendo la capacitación, la respuesta quimiotáctica y la exocitosis acrosomal. Pese a estas observaciones, los mecanismos a través de los cuales la progesterona estimula fenómenos tan diversos en el espermatozoide son aún desconocidos. Tampoco se conoce con exactitud el papel funcional y los mecanismos de acción de algunas proteínas del plasma seminal que interaccionan y se unen a los espermatozoides, con alta especificidad, durante la eyaculación. Por lo tanto, resulta altamente interesante profundizar los estudios sobre las propiedades funcionales de las proteínas caltrin (calcium transport inhibitor) y ß-microseminoprotein (MSP) del plasma seminal de mamíferos, las cuales responden a las características mencionadas. Los estudios hasta ahora realizados han dado cuenta de que caltrin inhibe la incorporación de Ca2+ extracelular, previene la exocitosis acrosomal espontánea y promueve la unión espermatozoide-zona pelúcida. También hay datos preliminares que sugieren un efecto inhibitorio sobre la movilidad hiperactivada de los espermatozoides. Respecto a MSP, sólo se sabe que inhibe la exocitosis acrosomal espontánea y que su contenido, en el plasma seminal, guarda una relación inversa con la fertilidad. Por todo lo expuesto, se propone estudiar los mecanismos de acción de la progesterona y las proteínas caltrin y MSP sobre los procesos fisiológicos antes indicados. Para ello, se estudiarán las variaciones de Ca2+ intracelular en espermatozoides individuales sometidos a diferentes tratamientos (gradientes de progesterona, capacitación en presencia y ausencia de caltrin y/o MSP, etc.), usando video microscopía de fluorescencia y análisis computarizado de imágenes. También se examinará la influencia de estas moléculas sobre la interacción de gametas y la fertilización.

Apoptosis y expresión de receptores de reconocimiento de patrones en Polimorfonucleares Neutrófilos de individuos con infección VIH/SIDA. Apoptosis and recognition patterns receptors expression in Polymorphonuclear Neutrophils of individuals with HIV/AIDS infection.

El Virus de la Inmunodeficiencia Humana tipo 1 (VIH-1) afecta principalmente a la respuesta inmune específica causando una pérdida progresiva de los linfocitos T CD4+. Sin embargo, este virus también afecta a células del sistema inmune innato, tales como los Polimorfonucleares Neutrófilos (PMN). Existen evidencias de alteraciones funcionales de los PMN durante la progresión de la infección por VIH y una de las explicaciones de estos defectos, la atribuye a una muerte celular programada o apoptosis constitutiva incrementada. El compromiso de la apoptosis de los PMN en la infección por VIH no está totalmente dilucidado, por ello, los objetivos de este proyecto son investigar el efecto de la infección por VIH sobre la apoptosis de PMN, analizar la expresión de moléculas y receptores de patrones de reconocimiento en estas células y evaluar el impacto de la terapia antirretroviral sobre la apoptosis y expresión de moléculas y receptores en PMN. Se incluirán individuos en distintos estadios clínicos e inmunológicos de la infección con o sin tratamiento antirretroviral y se determinarán parámetros hematológicos, inmunológicos y virológicos a fin de correlacionar el nivel de apoptosis y expresión de moléculas y receptores con el nivel de linfocitos T CD4+ y carga viral. La importancia de los PMN en el control de la infección por el VIH es actualmente un área de mucho interés, ya pueden ejercer un efecto anti-VIH directo, y al mismo tiempo, ser blancos de la infección viral. Los mecanismos que conducen a la muerte acelerada de los PMN no han sido totalmente dilucidados, por ello, su estudio permitirá entender las bases bioquímicas de los cambios morfológicos y determinar los mecanismos que definen su iniciación y regulación. En el presente proyecto, el estudio de la apoptosis de PMN de pacientes con infección VIH/SIDA posibilitará caracterizar la sobrevida de éstas células y su relación con el estado inmunológico, virológico y la terapia antirretroviral. Además, el estudio de los receptores reconocedores de patrones moleculares asociados a patógenos permitirá aclarar algunos aspectos de la activación de la respuesta inmune innata y su conexión con la inmunidad adaptativa. Comprender aspectos claves de la cascada de la apoptosis de PMN y de la expresión de receptores reconocedores de patrones moleculares en la infección VIH/SIDA podría en un futuro aportar posibles blancos terapéuticos para restaurar la función de estas células durante esta infección.

Producción in vitro de Embriones Bovinos Transgénicos mediante Inyección Intracitoplasmática de Espermatozoides (ICSI) con Enzimas de Restricción. In vitro Production of Transgenic Bovine Embryos using Intracitoplasmic Sperm Injection (ICSI) and Restriction Enzymes

La ingeniería genética y la reprogramación de organismos vivos representan las nuevas fronteras biotecnológicas que permitirán generar animales con modificaciones precisas en sus genomas para un sinnúmero de aplicaciones biomédicas y agropecuarias. Las técnicas para inducir modificaciones génicas intencionales en animales, especialmente en especies mayores de interés agropecuario, se encuentran rezagadas si se compara con los avances significativos que se han producido en el área de la transgénesis de roedores de laboratorio, especialmente el ratón. Es así que, el presente proyecto persigue desarrollar y optimizar protocolos para generar embriones bovinos transgénicos para aplicaciones biotecnológicas. La estrategia propuesta, se basa en conseguir la presencia simultánea en el interior celular de una enzima de restricción (I-SceI) más un transgén (formado por casetes de expresión de una proteína fluorescente -ZsGreen1- y neomicina fosfotransferasa). Específicamente, proyectamos estudiar una vía alternativa para generar embriones bovinos transgénicos mediante la incorporación del transgén (casetes ZsGreen1 y neo) flanqueado por sitios I-SceI más la enzima I-SceI al interior del ovocito junto con el espermatozoide durante la técnica conocida como inyección intracitoplasmática de espermatozoides (ICSI). Los embriones así generados se cultivarán in vitro, inspeccionándolos diariamente para detectar la emisión de fluorescencia, indicativa de la expresión de la proteína ZsGreen1. Los embriones que alcancen el estado de blastocisto y expresen el transgén se transferirán quirúrgicamente al útero de ovejas sincronizadas y se mantendrán durante 7 días. Al cabo de este período, los embriones se recolectarán quirúrgicamente del útero ovino y se transportarán al laboratorio para determinar el número de sitios de integración y número de copias del transgén mediante el análisis de su ADN por Southern blot. Se prevé que los resultados de esta investigación permitirán sentar las bases para el desarrollo de métodos eficientes para obtener modificaciones precisas en el genoma de los animales domésticos para futuras aplicaciones biotecnológicas. Genetic engineering and reprogrammed organisms represent the new biotechnological frontiers which will make possible to generate animals with precise genetic modifications for agricultural and biomedical applications. Current methods used to generate genetically modified large animals, lay behind those used in laboratory animals, specially the mouse. Therefore, we seek to develop and optimize protocols to produce transgenic bovine embryos through the use of a non-viral vector. The strategy involves the simultaneous presence inside the cell of a restriction enzyme (I-SceI) and a transgene (carrying cassettes for a fluorescent protein -ZsGreen1- and neomycin phosphotransferase) flanked by restriction sites for the endonuclease. We plan to develop an alternative approach to generate transgenic bovine embryos by coinjecting the transgene flanked by I-SceI restriction sites plus the enzyme I-SceI along with the spermatozoon during the technique known as intracytoplasmic sperm injection (ICSI). Embryos will be cultured in vitro and inspected daily with a fluorescence microscope to characterize transgene expression. Embryos that reach the blastocyst stage and express the transgene will be surgically transfer to the uterus of a synchronized ewe. After 7 days, the embryos will be flushed out the ovine uterus and transported to the laboratory to determine the number of integration sites and transgene copies by Southern blot. We anticipate that results from this research will set the stage for the development of efficient strategies to achieve precise genetic modifications in large domestic animals for future biotechnological applications.

Development of a semi-automatic self learning pattern recognition system for the control of electronic boards

This project was funded under the Applied Research Grants Scheme administered by Enterprise Ireland. The project was a partnership between Galway - Mayo Institute of Technology and an industrial company, Tyco/Mallinckrodt Galway. The project aimed to develop a semi - automatic, self - learning pattern recognition system capable of detecting defects on the printed circuits boards such as component vacancy, component misalignment, component orientation, component error, and component weld. The research was conducted in three directions: image acquisition, image filtering/recognition and software development. Image acquisition studied the process of forming and digitizing images and some fundamental aspects regarding the human visual perception. The importance of choosing the right camera and illumination system for a certain type of problem has been highlighted. Probably the most important step towards image recognition is image filtering, The filters are used to correct and enhance images in order to prepare them for recognition. Convolution, histogram equalisation, filters based on Boolean mathematics, noise reduction, edge detection, geometrical filters, cross-correlation filters and image compression are some examples of the filters that have been studied and successfully implemented in the software application. The software application developed during the research is customized in order to meet the requirements of the industrial partner. The application is able to analyze pictures, perform the filtering, build libraries, process images and generate log files. It incorporates most of the filters studied and together with the illumination system and the camera it provides a fully integrated framework able to analyze defects on printed circuit boards.

Emotion recognition within spoken dialog systems

Magdeburg, Univ., Fak. für Elektrotechnik und Informationstechnik, Diss., 2011

Neural mechanisms for fast recognition of auditory emotion

Emotion, audition, event-related potentials, MMN, multidimensional scaling, timbre, perception

Estimation of nonparametric probability density functions with applications to automatic speech recognition

Magdeburg, Univ., Fak. für Elektrotechnik und Informationstechnik, Diss., 2007

Forearm surface EMG signals recognition and muscoloskeletal system dynamics identification using intelligent computational methods

Biosignals processing, Biological Nonlinear and time-varying systems identification, Electomyograph signals recognition, Pattern classification, Fuzzy logic and neural networks methods

Discriminative classifiers for speaker recognition

Speaker Recognition, Speaker Verification, Sparse Kernel Logistic Regression, Support Vector Machine

Hand gesture spotting and recognition using HMMs and CRFs in color image sequences

Magdeburg, Univ., Fak. für Elektrotechnik und Informationstechnik, Diss., 2010

Bases para o estudo da genética de populações dos Hymenoptera em geral e dos Apinae sociais em particular

This paper deals with problems on population genetics in Hymenoptera and particularly in social Apidae. 1) The studies on populations of Hymenoptera were made according to the two basic types of reproduction: endogamy and panmixia. The populations of social Apinae have a mixed method of reproduction with higher percentage of panmixia and a lower of endogamy. This is shown by the following a) males can enter any hive in swarming time; b) males of Meliponini are expelled from hives which does not need them, and thus, are forced to look for some other place; c) Meliponini males were seen powdering themselves with pollen, thus becoming more acceptable in any other hive. The panmixia is not complete owing to the fact that the density of the breeding population as very low, even in the more frequent species as low as about 2 females and 160 males per reproductive area. We adopted as selection values (or survival indices) the expressions according to Brieger (1948,1950) which may be summarised as follows; a population: p2AA + ²pq Aa + q2aa became after selection: x p2AA + 2pq Aa + z q²aa. For alge-braics facilities Brieger divided the three selective values by y giving thus: x/y p2 AA + y/y 2 pq Aa + z/y q²aa. He called x/y of RA and z/y of Ra, that are survival or selective index, calculated in relation to the heterozygote. In our case all index were calculated in relation to the heterozygote, including the ones for haploid males; thus we have: RA surveval index of genotype AA Ra surveval index of genotype aa R'A surveval index of genotype A R'a surveval index of genotype a 1 surveval index of genotype Aa The index R'A ande R'a were equalized to RA and Ra, respectively, for facilities in the conclusions. 2) Panmitic populations of Hymenoptera, barring mutations, migrations and selection, should follow the Hardy-Weinberg law, thus all gens will be present in the population in the inicial frequency (see Graphifc 1). 3) Heterotic genes: If mutation for heterotic gene ( 1 > RA > Ra) occurs, an equilibrium will be reached in a population when: P = R A + Ra - 2R²a _____________ (9) 2(R A + Ra - R²A - R²a q = R A + Ra - 2R²A _____________ (10) 2(R A + Ra - R²A - R²a A heterotic gene in an hymenopteran population may be maintained without the aid of new mutation only if the survival index of the most viable mutant (RA) does not exced the limiting value given by the formula: R A = 1 + √1+Ra _________ 4 If RA has a value higher thah the one permitted by the formula, then only the more viable gene will remain present in the population (see Graphic 10). The only direct proof for heterotic genes in Hymenoptera was given by Mackensen and Roberts, who obtained offspring from Apis mellefera L. queens fertilized by their own sons. Such inbreeding resulted in a rapid loss of vigor the colony; inbred lines intercrossed gave a high hybrid vigor. Other fats correlated with the "heterosis" problem are; a) In a colony M. quadrifasciata Lep., which suffered severely from heat, the percentage of deths omong males was greater .than among females; b) Casteel and Phillips had shown that in their samples (Apis melifera L). the males had 7 times more abnormalities tian the workers (see Quadros IV to VIII); c) just after emerging the males have great variation, but the older ones show a variation equal to that of workers; d) The tongue lenght of males of Apis mellifera L., of Bombus rubicundus Smith (Quadro X), of Melipona marginata Lep. (Quadro XI), and of Melipona quadrifasciata Lep. Quadro IX, show greater variationthan that of workers of the respective species. If such variation were only caused by subviables genes a rapid increasse of homozigoty for the most viable alleles should be expected; then, these .wild populations, supposed to be in equilibrium, could .not show such variability among males. Thus we conclude that heterotic genes have a grat importance in these cases. 4) By means of mathematical models, we came to the conclusion tht isolating genes (Ra ^ Ra > 1), even in the case of mutations with more adaptability, have only the opor-tunity of survival when the population number is very low (thus the frequency of the gene in the breeding population will be large just after its appearence). A pair of such alleles can only remain present in a population when in border regions of two races or subspecies. For more details see Graphics 5 to 8. 5) Sex-limited genes affecting only females, are of great importance toHymenoptera, being subject to the same limits and formulas as diploid panmitic populations (see formulas 12 and 13). The following examples of these genes were given: a) caste-determining genes in the genus Melipona; b) genes permiting an easy response of females to differences in feeding in almost all social Hymenoptera; c) two genes, found in wild populations, one in Trigona (Plebéia) mosquito F. SMITH (quadro XII) and other in Melipona marginata marginata LEP. (Quadro XIII, colonies 76 and 56) showing sex-limited effects. Sex-limited genes affecting only males do not contribute to the plasticity or genie reserve in hymenopteran populations (see formula 14). 6) The factor time (life span) in Hymenoptera has a particular importance for heterotic genes. Supposing one year to be the time unit and a pair of heterotic genes with respective survival indice equal to RA = 0, 90 and Ra = 0,70 to be present; then if the life time of a population is either one or two years, only the more viable gene will remain present (see formula 11). If the species has a life time of three years, then both alleles will be maintained. Thus we conclude that in specis with long lif-time, the heterotic genes have more importance, and should be found more easily. 7) The colonies of social Hymenoptera behave as units in competition, thus in the studies of populations one must determine the survival index, of these units which may be subdivided in indice for egg-laying, for adaptive value of the queen, for working capacity of workers, etc. 8) A study of endogamic hymenopteran populations, reproduced by sister x brother mating (fig. 2), lead us to the following conclusions: a) without selection, a population, heterozygous for one pair of alleles, will consist after some generations (theoretically after an infinite number of generation) of females AA fecundated with males A and females aa fecundated with males a (see Quadro I). b) Even in endogamic population there is the theoretical possibility of the presence of heterotic genes, at equilibrium without the aid of new mutations (see Graphics 11 and 12), but the following! conditions must be satisfied: I - surveval index of both homozygotes (RA e Ra) should be below 0,75 (see Graphic 13); II - The most viable allele must riot exced the less viable one by more than is permited by the following formula (Pimentel Gomes 1950) (see Gra-fic 14) : 4 R5A + 8 Ra R4A - 4 Ra R³A (Ra - 1) R²A - - R²a (4 R²a + 4 Ra - 1) R A + 2 R³a < o Considering these two conditions, the existance of heterotic genes in endogamic populations of Hymenoptera \>ecames very improbable though not - impossible. 9) Genie mutation offects more hymenopteran than diploid populations. Thus we have for lethal genes in diploid populations: u = q2, and in Hymenoptera: u = s, being u the mutation ratio and s the frequency of the mutant in the male population. 10) Three factors, important to competition among species of Meliponini were analysed: flying capacity of workers, food gathering capacity of workers, egg-laying of the queen. In this connection we refer to the variability of the tongue lenght observed in colonies from several localites, to the method of transporting the pollen in the stomach, from some pots (Melliponi-ni storage alveolus) to others (e. g. in cases of pillage), and to the observation that the species with the most populous hives are almost always the most frequent ones also. 11) Several defensive ways used for Meliponini to avoid predation are cited, but special references are made upon the camouflage of both hive (fig. 5) and hive entrance (fig. 4) and on the mimetism (see list in page ). Also under the same heading we described the method of Lestrimelitta for pillage. 12) As mechanisms important for promoting genetic plasticity of hymenopteran species we cited: a) cytological variations and b) genie reserve. As to the former, duplications and numerical variations of chromosomes were studied. Diprion simile ATC was cited as example for polyploidy. Apis mellife-ra L. (n •= 16) also sugests polyploid origen since: a) The genus Melipona, which belongs to a" related tribe, presents in all species so far studied n = 9 chromosomes and b) there occurs formation of dyads in the firt spermatocyte division. It is su-gested that the origin of the sex-chromosome of Apis mellifera It. may be related to the possible origin of diplo-tetraploidy in this species. With regards to the genie reserve, several possible types of mutants were discussed. They were classified according to their survival indices; the heterotic and neutral mutants must be considered as more important for the genie reserve. 13) The mean radius from a mother to a daghter colony was estimated as 100 meters. Since the Meliponini hives swarm only once a year we may take 100 meters a year as the average dispersion of female Meliponini in ocordance to data obtained from Trigona (tetragonisca) jaty F. SMITH and Melipona marginata LEP., while other species may give different values. For males the flying distance was roughly estimated to be 10 times that for females. A review of the bibliography on Meliponini swarm was made (pg. 43 to 47) and new facts added. The population desity (breeding population) corresponds in may species of Meliponini to one male and one female per 10.000 square meters. Apparently the males are more frequent than the females, because there are sometimes many thousands, of males in a swarm; but for the genie frequency the individuals which have descendants are the ones computed. In the case of Apini and Meliponini, only one queen per hive and the males represented by. the spermatozoos in its spermateca are computed. In Meliponini only one male mate with the queen, while queens of Apis mellijera L. are fecundated by an average of about 1, 5 males. (Roberts, 1944). From the date cited, one clearly sees that, on the whole, populations of wild social bees (Meliponini) are so small that the Sewall Wright effect may become of great importance. In fact applying the Wright's formula: f = ( 1/aN♂ + 1/aN♀) (1 - 1/aN♂ + 1/aN♀) which measures the fixation and loss of genes per generation, we see that the fixation or loss of genes is of about 7% in the more frequent species, and rarer species about 11%. The variation in size, tergite color, background color, etc, of Melipona marginata Lep. is atributed to this genetic drift. A detail, important to the survival of Meliponini species, is the Constance of their breeding population. This Constance is due to the social organization, i. e., to the care given to the reproductive individuals (the queen with its sperm pack), to the way of swarming, to the food storage intended to control variations of feeding supply, etc. 14) Some species of the Meliponini are adapted to various ecological conditions and inhabit large geographical areas (e. g. T. (Tetragonisca jaty F. SMITH), and Trigona (Nanno-trigona testaceicornis LEP.) while others are limited to narrow regions with special ecological conditions (e. g. M. fuscata me-lanoventer SCHWARZ). Other species still, within the same geographical region, profit different ecological conditions, as do M. marginata LEP. and M. quadrifasciata LEP. The geographical distribution of Melipona quadrifasciata LEP. is different according to the subspecies: a) subsp anthidio-des LEP. (represented in Fig. 7 by black squares) inhabits a region fron the North of the S. Paulo State to Northeastern Brazil, ,b) subspecies quadrifasciata LEP., (marked in Fig. 7 with black triangles) accurs from the South of S. Paulo State to the middle of the State of Rio Grande do Sul (South Brazil). In the margined region between these two areas of distribution, hi-brid colonies were found (Fig. 7, white circles); they are shown with more details in fig. 8, while the zone of hybridization is roughly indicated in fig. 9 (gray zone). The subspecies quadrifasciata LEP., has 4 complete yellow bands on the abdominal tergites while anthidioides LEP. has interrupted ones. This character is determined by one or two genes and gives different adaptative properties to the subspecies. Figs. 10 shows certains meteorological isoclines which have aproximately the same configuration as the limits of the hybrid zone, suggesting different climatic adaptabilities for both genotypes. The exis-tance of a border zone between the areas of both subspecies, where were found a high frequency of hybrids, is explained as follows: being each subspecies adapted to a special climatic zone, we may suppose a poor adaptation of either one in the border region, which is also a region of intermediate climatic conditions. Thus, the hybrids, having a combination of the parent qualities, will be best adapted to the transition zone. Thus, the hybrids will become heterotic and an equilibrium will be reached with all genotypes present in the population in the border region.