IDENTIFICATION OF SUITABLE AREAS FOR IRRIGATED RICE CROPPING USING MODIS IMAGES AND HAND MODEL

ABSTRACT One of the most relevant activities of Brazilian economy is agriculture. Among the main crops in Brazil, rice is one of high relevance. The state of Rio Grande do Sul, in Southern Brazil, is responsible for 68.7% of domestic production (IBGE, 2013). The goal of this study was to develop a low-cost methodology with a regional scope to identify suitable areas for irrigated rice cropping in this state, using spectro-temporal behavior of vegetation index by means of MODIS images and HAND model. The rice-cropped area of this study was the southern half of the State. Using the HAND model, flood areas were mapped to identify irrigated rice cultivation. We used multi-temporal images of vegetation index from MODIS sensor, covering the period from August 2001 to May 2012. To assess the results, we used data collected in the fields and cropped area information from IBGE. The results showed that the proposed methodology was satisfactory, with Kappa 0.92 and global accuracy of 98.18%. As result, MODIS sensor data and flood areas delineation by means of HAND model generated the estimate irrigated rice area for the area of study.

Pluripotency and genetic stability of human pluripotent stem cells

Pluripotent cells have the potential to differentiate into all somatic cell types. As the adult human body is unable to regenerate various tissues, pluripotent cells provide an attractive source for regenerative medicine. Human embryonic stem cells (hESCs) can be isolated from blastocyst stage embryos and cultured in the laboratory environment. However, their use in regenerative medicine is restricted due to problems with immunosuppression by the host and ethical legislation. Recently, a new source of pluripotent cells was established via the direct reprogramming of somatic cells. These human induced pluripotent stem cells (hiPSCs) enable the production of patient specific cell types. However, numerous challenges, such as efficient reprogramming, optimal culture, directed differentiation, genetic stability and tumor risk need to be solved before the launch of therapeutic applications. The main objective of this thesis was to understand the unique properties of human pluripotent stem cells. The specific aims were to identify novel factors involved in maintaining pluripotency, characterize the effects of low oxygen culture on hESCs, and determine the high resolution changes in hESCs and hiPSCs during culture and reprogramming. As a result, the previously uncharacterized protein L1TD1 was determined to be specific for pluripotent cells and essential for the maintenance of pluripotency. The low oxygen culture supported undifferentiated growth and affected expression of stem cell associated transcripts. High resolution screening of hESCs identified a number of culture induced copy number variations and loss of heterozygosity changes. Further, screening of hiPSCs revealed that reprogramming induces high resolution alterations. The results obtained in this thesis have important implications for stem cell and cancer biology and the therapeutic potential of pluripotent cells.

Cardiorespiratory responses during and after water exercise in pregnant and non-pregnant women

PURPOSE: to compare the blood pressure and oxygen consumption (VO2) responses between pregnant and non-pregnant women, during cycle ergometer exercise on land and in water. METHODS: ten pregnant (27 to 29 weeks of gestation) and ten non-pregnant women were enrolled. Two cardiopulmonary tests were performed on a cycle ergometer (water and land) at the heart rate corresponding to VO2, over a period of 30 minutes each. Exercise measurements consisted of recording blood pressure every five minutes, and heart rate and VO2 every 20 seconds. Two-way ANOVA was used and α=0.05 (SPSS 17.0). RESULTS: there was no difference in cardiovascular responses between pregnant and non-pregnant women during the exercise. The Pregnant Group demonstrated significant differences in systolic (131.6±8.2; 142.6±11.3 mmHg), diastolic (64.8±5.9; 74.5±5.3 mmHg), and mean blood pressure (87.0±4.1; 97.2±5.7 mmHg), during water and land exercise, respectively. The Non-pregnant women Group also had a significantly lower systolic (130.5±8.4; 135.9±8.7 mmHg), diastolic (67.4±5.7; 69.0±10.1 mmHg), and mean blood pressure (88.4±4.8; 91.3±7.8 mmHg) during water exercise compared to the land one. There were no significant differences in VO2 values between water and land exercises or between pregnant and non-pregnant women. After the first five-minute recovery period, both blood pressure and VO2 were similar to pre-exercise values. CONCLUSIONS: for pregnant women with 27 to 29 weeks of gestation, water exercise at the heart rate corresponding to VO2 is physiologically appropriate. These women also present a lower blood pressure response to exercise in water than on land.

Effects of laparoscopy on the cardiorespiratory system of brown brocket deer (Mazama gouazoubira) anesthetized with ketamine/ xylazine/ midazolam combination and isoflurane

Laparoscopy is not widely used as a tool to perform assisted reproduction techniques in South American cervids; thus, scarce information in literature is available regarding its effects and appropriate anesthetic protocols to perform it. This study evaluated the effect of laparoscopy on heart rate (HR), respiration rate (RR), saturation of oxyhemoglobin (SpO2) and rectal temperature (RT) of six female brown brocket deer (Mazama gouazoubira) anesthetized with ketamine (5mg/kg), xylazine (0.3mg/kg), midazolam (0.5mg/kg) combination i.v. and isoflurane. Twelve laparoscopies were performed and each animal was used twice with a 40-day interval. After anesthetized, the animals were placed in dorsal recumbency to perform laparoscopy procedure using abdominal CO2 insufflations (14.2 ± 2.39mmHg; M ± SE). The main events of the laparoscopy procedure were divided into three periods: animal without (P1) and with abdominal insufflation (P2) and abdominal insufflation with the hips raised at 45º (P3). As a control, the animals were anesthetized again 40 days after the last laparoscopy, and were maintained in a dorsal recumbency for the same average duration of the previous anesthesia and no laparoscopy procedure was conducted. The period of anesthesia for the controls was also divided into P1, P2, and P3 considering the average duration of these periods in previous laparoscopies performed. Data were analyzed through the (ANOVA) variance analysis followed by Tukey test and values at P<0.05 were considered significant. No significant differences were observed in the parameters evaluated at P1, P2 and P3 between the animals submitted to laparoscopy and control. However, the RR mean between P1 (38.8 ± 4.42) and P3 (32.7 ± 4.81); and the RT mean between the P1 (38.2ºC ± 0.17), P2 (37.6ºC ± 0.19) and P3 (37.0ºC ± 0.21) varied significantly, independent of the laparoscopy. These data indicated that laparoscopy didn't cause any significant alterations in the cardiorespiratory parameters evaluated, even though the anesthetic protocol used can cause a reduction in the RT contributing to development of hypothermia during anesthesia.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Ultrastructural characterization of bovine umbilical cord blood cells

The umbilical cord blood (UCB) is an important source of pluripotent stem cells, which motivated researches on ontogeny and transplantation. The morphological characterization of umbilical cord cells is the first step to establish subsequent experiments on these areas. Although some information on humans can be found, no data on UCB is available for bovines. Therefore, this work is the first attempt to conduct an ultrastructural characterization of bovine umbilical cord blood. Blood was collected from the umbilical cord of twenty fetuses by punction of the umbilical vein. Samples were processed for whole leucocytes observation by centrifugation and the buffy coat was collected. Cells were washed and pelleted and prepared according to the standard protocol of the transmission electron microscopy. The presence of cells with morphologic characteristics compatible with the precursors from the erythrocytic, neutrophilic, eosinophilic, basophilic, and lymphocytic lineages was observed. Atypical cells with peculiar morphological features, strongly similar to apoptotic cells, were seen. Bovine neutrophils with three types of cytoplasmic granules were also found in the blood. The ultrastructural characteristics of observed bovine UCB cells where similar to those found in other species, suggesting that bovines could possibly constitute an experimental model for approaches on UCB cells research.

A global transport packaging guideline for selected business areas

In this Master’s Thesis a global transport packaging guideline for selected business areas was compiled for the Fiskars the company, which provides branded consumer goods for home, garden and outdoor use. The business areas included were Home and Garden business areas. The aim of the guideline was to be a comprehensive guide for the suppliers, product development, operations and external vendors of the company. The guideline consists of written instructions, tables and illustrations that provide useful information for players working with transport packages from sourcing through to shipments. As the role of corporate responsibility and sustainability has grown, a part of responsible manufacturing strategy includes using materials that are re-usable, recyclable or recoverable as energy or through composting. Hence packaging waste management implementations of different regions were also inspected. The resulting guide covers a range of topics concerning packaging and its transport. The topics include legal requirements, restricted materials and substances, preferred materials, markings, labeling of boxes, logistics and distribution center requirements, physical testing and an inspection checklist.

Exploring Customer-Oriented Service Offerings in Public Health: Developing Integrated Health Care Services for Rural Areas

Väestön ikääntyminen pakottaa yhteiskunnan ja julkisen terveydenhuollon muutoksiin. Jotta ikääntyvien ihmisten kotona asuminen voidaan mahdollistaa, palvelujärjestelmän pitää mukautua muuttuvaan tilanteeseen. Tämän diplomityön tarkoituksena on tunnistaa asiakaslähtöisiä lähellä asiakasta tarjottavia palvelukokonaisuuksia. Tutkimuksen teoreettinen viitekehys muodostuu asiakasarvon luomisesta ja palvelutarjoamista. Tarkasteluryhmänä on Etelä-Karjalan alueen 60–90-vuotiaat ja käytetty aineisto on kerätty vastaajilta postitse lähetetyllä kyselyllä. Tutkimus on eksploratiivinen ja tulosten tulkinnassa on hyödynnetty määrällisen tutkimuksen ja verkostoanalyysin menetelmiä. Työn keskeisimmät tulokset ovat tunnistetut asiakassegmentit ja heidän tarpeidensa pohjalta muodostetut palvelupaketit. Tulokset indikoivat asiakkaiden tarpeita ja tuloksia on analysoitu myös tuottajan näkökulmasta. Empiiristen tulosten lisäksi teoriaviitekehystä on kehitetty eteenpäin, jotta palvelukeskeiset teoriat voidaan ymmärtää yritysten näkökulman lisäksi asiakkaan näkökulmasta.

Regulation of self-renewal and detection of karyotypic changes of pluripotent human embryonic stem cells

Human embryonic stem cells are pluripotent cells capable of renewing themselves and differentiating to specialized cell types. Because of their unique regenerative potential, pluripotent cells offer new opportunities for disease modeling, development of regenerative therapies, and treating diseases. Before pluripotent cells can be used in any therapeutic applications, there are numerous challenges to overcome. For instance, the key regulators of pluripotency need to be clarified. In addition, long term culture of pluripotent cells is associated with the accumulation of karyotypic abnormalities, which is a concern regarding the safe use of the cells for therapeutic purposes. The goal of the work presented in this thesis was to identify new factors involved in the maintenance of pluripotency, and to further characterize molecular mechanisms of selected candidate genes. Furthermore, we aimed to set up a new method for analyzing genomic integrity of pluripotent cells. The experimental design applied in this study involved a wide range of molecular biology, genome-wide, and computational techniques to study the pluripotency of stem cells and the functions of the target genes. In collaboration with instrument and reagent company Perkin Elmer, KaryoliteTM BoBsTM was implemented for detecting karyotypic changes of pluripotent cells. Novel genes were identified that are highly and specifically expressed in hES cells. Of these genes, L1TD1 and POLR3G were chosen for further investigation. The results revealed that both of these factors are vital for the maintenance of pluripotency and self-renewal of the hESCs. KaryoliteTM BoBsTM was validated as a novel method to detect karyotypic abnormalities in pluripotent stem cells. The results presented in this thesis offer significant new information on the regulatory networks associated with pluripotency. The results will facilitate in understanding developmental and cancer biology, as well as creating stem cell based applications. KaryoliteTM BoBsTM provides rapid, high-throughput, and cost-efficient tool for screening of human pluripotent cell cultures.

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

Mesenchymal stem cells (MSC) are increasingly being proposed as a therapeutic option for treatment of a variety of different diseases in human and veterinary medicine. Stem cells have been isolated from feline bone marrow, however, very few data exist about the morphology of these cells and no data were found about the morphometry of feline bone marrow-derived MSCs (BM-MSCs). The objectives of this study were the isolation, growth evaluation, differentiation potential and characterization of feline BM-MSCs by their morphological and morphometric characteristics. in vitro differentiation assays were conducted to confirm the multipotency of feline MSC, as assessed by their ability to differentiate into three cell lineages (osteoblasts, chondrocytes, and adipocytes). To evaluate morphological and morphometric characteristics the cells are maintained in culture. Cells were observed with light microscope, with association of dyes, and they were measured at 24, 48, 72 and 120h of culture (P1 and P3). The non-parametric ANOVA test for independent samples was performed and the means were compared by Tukey's test. On average, the number of mononuclear cells obtained was 12.29 (±6.05x10(6)) cells/mL of bone marrow. Morphologically, BM-MSCs were long and fusiforms, and squamous with abundant cytoplasm. In the morphometric study of the cells, it was observed a significant increase in average length of cells during the first passage. The cell lengths were 106.97±38.16µm and 177.91±71.61µm, respectively, at first and third passages (24 h). The cell widths were 30.79±16.75 µm and 40.18±20.46µm, respectively, at first and third passages (24 h).The nucleus length of the feline BM-MSCs at P1 increased from 16.28µm (24h) to 21.29µm (120h). However, at P3, the nucleus length was 26.35µm (24h) and 25.22µm (120h). This information could be important for future application and use of feline BM-MSCs.