A histopathologic evaluation of gross lesions excised from commercially important North Altantic marine fishes

Histopathologic studies of lesions found in commercially important North Atlantic marine fishes are uncommon. As part of a comprehensive Northeast Fisheries Center program ("Ocean Pulse") to evaluate environmental and resource health on the U.S. Continental Shelf from Cape Hatteras to Nova Scotia, grossly visible lesions of the gills, integument, muscle, and viscera of primarily bottom-dwelling fishes were excised and examined using light microscopy. Several gadid and pleuronectid fishes accounted for most of the lesions observed. Most pathological examinations were incidental to samples taken for age and growth determination and evaluation of predator/prey relationships. Several gadids, with either gill, heart, or spleen lesions, were sampled more intensively. Gill lesions principally affected gadids and were caused by either microsporidans or an unidentified oocyte-like cell. The majority of gastrointestinal lesions consisted of encapsulated or encysted larval worms or microsporidan-induced cysts. Few heart lesions were found. Integumental lesioos included ulcers, lymphocystis, and trematode metacercariae. Liver lesions almost always consisted of encapsulated or encysted larval helminths. Necrotic granulomata were seen in muscle and microsporidan-induced granulomata in spleen. Although not numerous, histologically interesting lesions were noted in integument, heart, liver, spleen, and muscle of several fish species. Histologic study of tissues excised from a variety of demersal and pelagic fishes from the eastern North Atlantic (France, Germany, Spain) revealed assorted integumental, renal, hepatic, and splenic lesions. Small sample size and non-random sampling precluded obtaining a meaningful quantitative estimate of the prevalence of the observed lesions in the population at risk; however, a useful census has been made of the types of lesions present in commercially important marine fishes. (PDF file contains 20 pages.)

Water quality assessment and characterisation of chlorophyll-a variability related to river discharges, within the southeastern Bay of Biscay : evaluation and development of chlorophyll-a algorithms for MODIS and MERIS imagery

222 p. : il.

Use of generalised additive models to categorise continuous variables in clinical prediction

13 P.

An evaluation of length-frequency sampling procedures and subsequent data analysis for purse seine-caught yellowfin tuna in the Eastern Pacific Ocean

ENGLISH: A two-stage sampling design is used to estimate the variances of the numbers of yellowfin in different age groups caught in the eastern Pacific Ocean. For purse seiners, the primary sampling unit (n) is a brine well containing fish from a month-area stratum; the number of fish lengths (m) measured from each well are the secondary units. The fish cannot be selected at random from the wells because of practical limitations. The effects of different sampling methods and other factors on the reliability and precision of statistics derived from the length-frequency data were therefore examined. Modifications are recommended where necessary. Lengths of fish measured during the unloading of six test wells revealed two forms of inherent size stratification: 1) short-term disruptions of existing pattern of sizes, and 2) transition zones between long-term trends in sizes. To some degree, all wells exhibited cyclic changes in mean size and variance during unloading. In half of the wells, it was observed that size selection by the unloaders induced a change in mean size. As a result of stratification, the sequence of sizes removed from all wells was non-random, regardless of whether a well contained fish from a single set or from more than one set. The number of modal sizes in a well was not related to the number of sets. In an additional well composed of fish from several sets, an experiment on vertical mixing indicated that a representative sample of the contents may be restricted to the bottom half of the well. The contents of the test wells were used to generate 25 simulated wells and to compare the results of three sampling methods applied to them. The methods were: (1) random sampling (also used as a standard), (2) protracted sampling, in which the selection process was extended over a large portion of a well, and (3) measuring fish consecutively during removal from the well. Repeated sampling by each method and different combinations indicated that, because the principal source of size variation occurred among primary units, increasing n was the most effective way to reduce the variance estimates of both the age-group sizes and the total number of fish in the landings. Protracted sampling largely circumvented the effects of size stratification, and its performance was essentially comparable to that of random sampling. Sampling by this method is recommended. Consecutive-fish sampling produced more biased estimates with greater variances. Analysis of the 1988 length-frequency samples indicated that, for age groups that appear most frequently in the catch, a minimum sampling frequency of one primary unit in six for each month-area stratum would reduce the coefficients of variation (CV) of their size estimates to approximately 10 percent or less. Additional stratification of samples by set type, rather than month-area alone, further reduced the CV's of scarce age groups, such as the recruits, and potentially improved their accuracy. The CV's of recruitment estimates for completely-fished cohorts during the 198184 period were in the vicinity of 3 to 8 percent. Recruitment estimates and their variances were also relatively insensitive to changes in the individual quarterly catches and variances, respectively, of which they were composed. SPANISH: Se usa un diseño de muestreo de dos etapas para estimar las varianzas de los números de aletas amari11as en distintos grupos de edad capturados en el Océano Pacifico oriental. Para barcos cerqueros, la unidad primaria de muestreo (n) es una bodega de salmuera que contenía peces de un estrato de mes-área; el numero de ta11as de peces (m) medidas de cada bodega es la unidad secundaria. Limitaciones de carácter practico impiden la selección aleatoria de peces de las bodegas. Por 10 tanto, fueron examinados los efectos de distintos métodos de muestreo y otros factores sobre la confiabilidad y precisión de las estadísticas derivadas de los datos de frecuencia de ta11a. Se recomiendan modificaciones donde sean necesarias. Las ta11as de peces medidas durante la descarga de seis bodegas de prueba revelaron dos formas de estratificación inherente por ta11a: 1) perturbaciones a corto plazo en la pauta de ta11as existente, y 2) zonas de transición entre las tendencias a largo plazo en las ta11as. En cierto grado, todas las bodegas mostraron cambios cíclicos en ta11a media y varianza durante la descarga. En la mitad de las bodegas, se observo que selección por ta11a por los descargadores indujo un cambio en la ta11a media. Como resultado de la estratificación, la secuencia de ta11as sacadas de todas las bodegas no fue aleatoria, sin considerar si una bodega contenía peces de un solo lance 0 de mas de uno. El numero de ta11as modales en una bodega no estaba relacionado al numero de lances. En una bodega adicional compuesta de peces de varios lances, un experimento de mezcla vertical indico que una muestra representativa del contenido podría estar limitada a la mitad inferior de la bodega. Se uso el contenido de las bodegas de prueba para generar 25 bodegas simuladas y comparar los resultados de tres métodos de muestreo aplicados a estas. Los métodos fueron: (1) muestreo aleatorio (usado también como norma), (2) muestreo extendido, en el cual el proceso de selección fue extendido sobre una porción grande de una bodega, y (3) medición consecutiva de peces durante la descarga de la bodega. EI muestreo repetido con cada método y distintas combinaciones de n y m indico que, puesto que la fuente principal de variación de ta11a ocurría entre las unidades primarias, aumentar n fue la manera mas eficaz de reducir las estimaciones de la varianza de las ta11as de los grupos de edad y el numero total de peces en los desembarcos. El muestreo extendido evito mayormente los efectos de la estratificación por ta11a, y su desempeño fue esencialmente comparable a aquel del muestreo aleatorio. Se recomienda muestrear con este método. El muestreo de peces consecutivos produjo estimaciones mas sesgadas con mayores varianzas. Un análisis de las muestras de frecuencia de ta11a de 1988 indico que, para los grupos de edad que aparecen con mayor frecuencia en la captura, una frecuencia de muestreo minima de una unidad primaria de cada seis para cada estrato de mes-área reduciría los coeficientes de variación (CV) de las estimaciones de ta11a correspondientes a aproximadamente 10% 0 menos. Una estratificación adicional de las muestras por tipo de lance, y no solamente mes-área, redujo aun mas los CV de los grupos de edad escasos, tales como los reclutas, y mejoró potencialmente su precisión. Los CV de las estimaciones del reclutamiento para las cohortes completamente pescadas durante 1981-1984 fueron alrededor de 3-8%. Las estimaciones del reclutamiento y sus varianzas fueron también relativamente insensibles a cambios en las capturas de trimestres individuales y las varianzas, respectivamente, de las cuales fueron derivadas. (PDF contains 70 pages)

EVALUATION OF FILM-SUBSTRATE ADHESION VIA IMPACT USING COATED BULLETS

A method was devised to evaluate the adhesion between a film and a substrate. A front-end coated bullet is accelerated by a gas gun and hits the substrate of the specimen under test. The impact generates a compressive stress pulse that propagates toward the film. After transmission through the interface, part of the pulse is reflected on the free surface of the film, and tensile stress arises at the film-substrate interface, possibly inducing debonding of the film. This dynamic process was demonstrated analytically and simulated numerically by the finite element method. The results validate the initial concept and lay the foundation for further optimization of this method.

Single cell proteomics microchip to profile immune function, with applications in stem cell biology, translational disease mechanism study and clinical therapeutics monitoring

In response to infection or tissue dysfunction, immune cells develop into highly heterogeneous repertoires with diverse functions. Capturing the full spectrum of these functions requires analysis of large numbers of effector molecules from single cells. However, currently only 3-5 functional proteins can be measured from single cells. We developed a single cell functional proteomics approach that integrates a microchip platform with multiplex cell purification. This approach can quantitate 20 proteins from >5,000 phenotypically pure single cells simultaneously. With a 1-million fold miniaturization, the system can detect down to ~100 molecules and requires only ~104 cells. Single cell functional proteomic analysis finds broad applications in basic, translational and clinical studies. In the three studies conducted, it yielded critical insights for understanding clinical cancer immunotherapy, inflammatory bowel disease (IBD) mechanism and hematopoietic stem cell (HSC) biology.

To study phenotypically defined cell populations, single cell barcode microchips were coupled with upstream multiplex cell purification based on up to 11 parameters. Statistical algorithms were developed to process and model the high dimensional readouts. This analysis evaluates rare cells and is versatile for various cells and proteins. (1) We conducted an immune monitoring study of a phase 2 cancer cellular immunotherapy clinical trial that used T-cell receptor (TCR) transgenic T cells as major therapeutics to treat metastatic melanoma. We evaluated the functional proteome of 4 antigen-specific, phenotypically defined T cell populations from peripheral blood of 3 patients across 8 time points. (2) Natural killer (NK) cells can play a protective role in chronic inflammation and their surface receptor – killer immunoglobulin-like receptor (KIR) – has been identified as a risk factor of IBD. We compared the functional behavior of NK cells that had differential KIR expressions. These NK cells were retrieved from the blood of 12 patients with different genetic backgrounds. (3) HSCs are the progenitors of immune cells and are thought to have no immediate functional capacity against pathogen. However, recent studies identified expression of Toll-like receptors (TLRs) on HSCs. We studied the functional capacity of HSCs upon TLR activation. The comparison of HSCs from wild-type mice against those from genetics knock-out mouse models elucidates the responding signaling pathway.

In all three cases, we observed profound functional heterogeneity within phenotypically defined cells. Polyfunctional cells that conduct multiple functions also produce those proteins in large amounts. They dominate the immune response. In the cancer immunotherapy, the strong cytotoxic and antitumor functions from transgenic TCR T cells contributed to a ~30% tumor reduction immediately after the therapy. However, this infused immune response disappeared within 2-3 weeks. Later on, some patients gained a second antitumor response, consisted of the emergence of endogenous antitumor cytotoxic T cells and their production of multiple antitumor functions. These patients showed more effective long-term tumor control. In the IBD mechanism study, we noticed that, compared with others, NK cells expressing KIR2DL3 receptor secreted a large array of effector proteins, such as TNF-α, CCLs and CXCLs. The functions from these cells regulated disease-contributing cells and protected host tissues. Their existence correlated with IBD disease susceptibility. In the HSC study, the HSCs exhibited functional capacity by producing TNF-α, IL-6 and GM-CSF. TLR stimulation activated the NF-κB signaling in HSCs. Single cell functional proteome contains rich information that is independent from the genome and transcriptome. In all three cases, functional proteomic evaluation uncovered critical biological insights that would not be resolved otherwise. The integrated single cell functional proteomic analysis constructed a detail kinetic picture of the immune response that took place during the clinical cancer immunotherapy. It revealed concrete functional evidence that connected genetics to IBD disease susceptibility. Further, it provided predictors that correlated with clinical responses and pathogenic outcomes.

Evaluation of woman fisherfolks involvement in fisheries activities in Kainji Lake basin

The extent of rural women fisherfolks contribution to fisheries was grossly under estimated and certainly under valued. The basis of their involvement in fisheries activities is to make themselves equal partners to men productive and self reliant participants in the process of improving their own and their family living standard and to enable them realize their full potentials. This survey is informed a system of data collection with the aid of questionnaires and analysis. Ninety-six of the questionnaires were administered to women fisher folks in eight fishing communities selected at random. Thirty seven (97) percent of the women fisher folks are between the ages of 25-35 years, 59.4% of them can only read and write in Arabic language and 21.9% only are literature in western education, which is a general characteristic of the rural population in Nigeria. 24.0% of the respondents are in full time fishing activities while 65-5% are fishmongers. They belong to cooperative societies but only 26% indicated to have benefited from loan and credit facilities. 84.4% of the respondents are sustained by the business. The major problems facing women fisherfolks includes poor market price, fish spoilage, high transport cost and lack of access to loan and credit facilities and extension assistance. Solution to these problems will increase their status benefit and development

Wind-driven desertification: process modeling, remote monitoring, and forecasting

Arid and semiarid landscapes comprise nearly a third of the Earth's total land surface. These areas are coming under increasing land use pressures. Despite their low productivity these lands are not barren. Rather, they consist of fragile ecosystems vulnerable to anthropogenic disturbance.

The purpose of this thesis is threefold: (I) to develop and test a process model of wind-driven desertification, (II) to evaluate next-generation process-relevant remote monitoring strategies for use in arid and semiarid regions, and (III) to identify elements for effective management of the world's drylands.

In developing the process model of wind-driven desertification in arid and semiarid lands, field, remote sensing, and modeling observations from a degraded Mojave Desert shrubland are used. This model focuses on aeolian removal and transport of dust, sand, and litter as the primary mechanisms of degradation: killing plants by burial and abrasion, interrupting natural processes of nutrient accumulation, and allowing the loss of soil resources by abiotic transport. This model is tested in field sampling experiments at two sites and is extended by Fourier Transform and geostatistical analysis of high-resolution imagery from one site.

Next, the use of hyperspectral remote sensing data is evaluated as a substantive input to dryland remote monitoring strategies. In particular, the efficacy of spectral mixture analysis (SMA) in discriminating vegetation and soil types and detennining vegetation cover is investigated. The results indicate that hyperspectral data may be less useful than often thought in determining vegetation parameters. Its usefulness in determining soil parameters, however, may be leveraged by developing simple multispectral classification tools that can be used to monitor desertification.

Finally, the elements required for effective monitoring and management of arid and semiarid lands are discussed. Several large-scale multi-site field experiments are proposed to clarify the role of wind as a landscape and degradation process in dry lands. The role of remote sensing in monitoring the world's drylands is discussed in terms of optimal remote sensing platform characteristics and surface phenomena which may be monitored in order to identify areas at risk of desertification. A desertification indicator is proposed that unifies consideration of environmental and human variables.

Functional evaluation of noncovalent interactions in neuroreceptors and progress toward the expansion of unnatural amino acid methodology

This dissertation primarily describes chemical-scale studies of G protein-coupled receptors and Cys-loop ligand-gated ion channels to better understand ligand binding interactions and the mechanism of channel activation using recently published crystal structures as a guide. These studies employ the use of unnatural amino acid mutagenesis and electrophysiology to measure subtle changes in receptor function.

In chapter 2, the role of a conserved aromatic microdomain predicted in the D3 dopamine receptor is probed in the closely related D2 and D4 dopamine receptors. This domain was found to act as a structural unit near the ligand binding site that is important for receptor function. The domain consists of several functionally important noncovalent interactions including hydrogen bond, aromatic-aromatic, and sulfur-π interactions that show strong couplings by mutant cycle analysis. We also assign an alternate interpretation for the linear fluorination plot observed at W6.48, a residue previously thought to participate in a cation-π interaction with dopamine.

Chapter 3 outlines attempts to incorporate chemically synthesized and in vitro acylated unnatural amino acids into mammalian cells. While our attempts were not successful, method optimizations and data for nonsense suppression with an in vivo acylated tRNA are included. This chapter is aimed to aid future researchers attempting unnatural amino acid mutagenesis in mammalian cells.

Chapter 4 identifies a cation-π interaction between glutamate and a tyrosine residue on loop C in the GluClβ receptor. Using the recently published crystal structure of the homologous GluClα receptor, other ligand-binding and protein-protein interactions are probed to determine the similarity between this invertebrate receptor and other more distantly related vertebrate Cys-loop receptors. We find that many of the interactions previously observed are conserved in the GluCl receptors, however care must be taken when extrapolating structural data.

Chapter 5 examines inherent properties of the GluClα receptor that are responsible for the observed glutamate insensitivity of the receptor. Chimera synthesis and mutagenesis reveal the C-terminal portion of the M4 helix and the C-terminus as contributing to formation of the decoupled state, where ligand binding is incapable of triggering channel gating. Receptor mutagenesis was unable to identify single residue mismatches or impaired protein-protein interactions within this domain. We conclude that M4 helix structure and/or membrane dynamics are likely the cause of ligand insensitivity in this receptor and that the M4 helix has an role important in the activation process.

Evaluation of Two Alternative Carbon Capture and Storage Technologies: A Stochastic Model

30 p.

The application of methods of ecosystem analysis to the evaluation of water quality

Guided by experience and the theoretical development of hydrobiology, it can be considered that the main aim of water quality control should be the establishment of the rates of the self-purification process of water bodies which are capable of maintaining communities in a state of dynamic balance without changing the integrity of the ecosystem. Hence, general approaches in the elaboration of methods for hydrobiological control are based on the following principles: a. the balance of matter and energy in water bodies; b. the integrity of the ecosystem structure and of its separate components at all levels. Ecosystem analysis makes possible a revelation of the whole totality of factors which determine the anthropogenic evolution of a water body. This is necessary for the study of long-term changes in water bodies. The principles of ecosystem analysis of water bodies, together with the creation of their mathematical models, are important because, in future, with the transition of water demanding production into closed cycles of water supply, changes in water bodies will arise in the main through the influence of 'diffuse' pollution (from the atmosphere, with utilisation in transport etc.).

Optimization of the pyrolysis process for the production of a biomass derived reducing agent and hydrogen-rich gases

221 p.

Mechanisms involved in Escherichia coli and Serratia marcescens removal during activated sludge wastewater treatment

Wastewater treatment reduces environmental contamination by removing gross solids and mitigating the effects of pollution. Treatment also reduces the number of indicator organisms and pathogens. In this work, the fates of two coliform bacteria, Escherichia coli and Serratia marcescens, were analyzed in an activated sludge process to determine the main mechanisms involved in the reduction of pathogenic microorganisms during wastewater treatment. These bacteria, modified to express green fluorescent protein, were inoculated in an activated sludge unit and in batch systems containing wastewater. The results suggested that, among the different biological factors implied in bacterial removal, bacterivorous protozoa play a key role. Moreover, a representative number of bacteria persisted in the system as free-living or embedded cells, but their distribution into liquid or solid fractions varied depending on the bacterium tested, questioning the real value of bacterial indicators for the control of wastewater treatment process. Additionally, viable but nonculturable cells constituted an important part of the bacterial population adhered to solid fractions, what can be derived from the competition relationships with native bacteria, present in high densities in this environment. These facts, taken together, emphasize the need for reliable quantitative and qualitative analysis tools for the evaluation of pathogenic microbial composition in sludge, which could represent an undefined risk to public health and ecosystem functions when considering its recycling.

Final Evaluation Memorandum: Strategies for Resolving Low Dissolved Oxygen and Methylmercury Events in Northern Suisun Marsh

The purpose of the project is to improve our understanding about best management practices that can be utilized on diked managed wetlands in Suisun Marsh for reducing the occurrence of low dissolved oxygen (DO) and high methylmercury (MeHg) events associated primarily with fall flood-up practices. Low DO events are of concern because they can lead to undue stress and even mortality of sensitive aquatic organisms. Elevated MeHg levels are of concern because MeHg is a neurotoxin that bio-magnifies up the food chain and can cause deleterious effects to higher trophic level consumers such as piscivorous fish, birds, and mammals (including humans). This study involved two years (2007-2008) of intensive field data collection at two managed wetland sites in northwest Suisun Marsh and their surrounding tidal sloughs, an area with prior documented low DO events. In addition, the study collected limited soils and water quality field data and mapped vegetation for three managed wetland sites in the central interior of Suisun Marsh, for the purpose of examining whether wetlands at other locations exhibit characteristics that could indicate potential for similar concerns. In Year 1 of the study, the objective was to identify the baseline conditions in the managed wetlands and determine which physical management conditions could be modified for Year 2 to reduce low DO and MeHg production issues most effectively. The objective of Year 2 was to evaluate the effectiveness of these modified management actions at reducing production of low DO and elevated MeHg conditions within the managed wetlands and to continue improving understanding of the underlying biogeochemical processes at play. This Final Evaluation Memorandum examined a total of 19 BMPs, 14 involving modified water management operations and the remaining five involving modified soil and vegetation management practices. Some of these BMPs were previously employed and others have not yet been tested. For each BMP this report assesses its efficacy in improving water quality conditions and potential conflicts with wetland management. It makes recommendations for further study (either feasibility assessments or field testing) and whether to consider for future use. Certain previously used BMPs were found to be important contributors to poor water quality conditions and their continued use is not recommended. Some BMPs that could improve water quality conditions appear difficult to implement in regards to compatibility with wetland management; these BMPs require further elaboration and feasibility assessment to determine whether they should be field tested. In practice for any given wetland, there is likely a combination of BMPs that would together have the greatest potential to address the low DO and high MeHg water quality concerns. Consequently, this report makes no sweeping recommendations applicable to large groups of wetlands but instead promotes a careful consideration of factors at each wetland or small groups of wetlands and from that assessment to apply the most effective suite of BMPs. This report also identifies a number of recommended future actions and studies. These recommendations are geared toward improving the process understanding of factors that promote low DO and high MeHg conditions, the extent of these problems in Suisun Marsh, the regulatory basis for the DO standards for a large estuarine marsh, the economics of BMPs, and alternative approaches to BMPs on diked managed wetlands that may address the water quality issues. The most important of these recommendations is that future BMP implementation should be carried out within the context of rigorous scientific evaluation so as to gain the maximum improvement in how to manage these water quality issues in the diked managed wetlands of Suisun Marsh.

Análise de custo de um programa de prevenção de internação para idosos no ambiente da saúde suplementar

Esta tese realiza uma análise de custo de um programa de prevenção de internação hospitalar para idosos (acima de 65 anos), no ambiente da saúde suplementar. Trata-se um programa voltado para idosos considerados de risco, de acordo com um questionário que avalia a probabilidade de risco de internação hospitalar. Indivíduos de risco são convidados a participar do programa, quando se estabelecem dois grupos: um que aceita a intervenção do programa (Grupo Bem Viver) e outro que recusa (Grupo Recusa). A intervenção do programa está voltada para o acompanhamento domiciliar, com equipe multidisciplinar, que se efetiva de acordo com uma estratificação, segundo a avaliação da dependência funcional para os indivíduos. Essa estratificação funcional divide os idosos do programa em cinco grupos distintos, denominados grupos funcionais. Indivíduos que recusam a intervenção mantêm a assistência com cuidado usual. A análise de custo é uma avaliação econômica parcial que estabelece uma comparação da utilização de serviços de saúde entre os dois grupos de idosos: um sob intervenção, grupo Bem Viver, e outro em cuidado usual, grupo Recusa. Esta análise foi realizada para um período dividido em 12 meses antes do evento (intervenção ou recusa) e 12 meses após o evento. Levantou-se a utilização de serviços de saúde de acordo com os indicadores de consultas, exames, procedimentos ambulatoriais, internações e consultas hospitalares. O custo do programa foi incluído também na análise final. Comparações foram estabelecidas entre os grupos no padrão de utilização, considerando-se os períodos antes e depois do evento. Os grupos funcionais foram analisados dentro da perspectiva da utilização para os mesmos períodos. Os resultados demonstraram que o grupo com adesão tem um grande percentual de indivíduos com dependência funcional, sugerindo que existe uma seleção adversa na aceitação do programa, quando indivíduos de maior risco têm adesão. Para o grupo sob intervenção, ocorreu um aumento da utilização com os serviços de saúde no conjunto de consultas, exames e procedimentos ambulatoriais, havendo diminuição com o custo unitário das internações hospitalares e consultas hospitalares. O grupo que recusa tem discreto aumento para o conjunto de consultas, exames e procedimentos ambulatoriais e um grande percentual de aumento para as internações (+40.2%) e consultas hospitalares (34.3%). Este resultado para internações foi analisado também através da transformação logarítmica, com uso da função inversa para a base de cálculo. Nesta aplicação estatística foi confirmado esse resultado para o padrão de utilização de internações. Na análise final, que incluiu o custo do programa propriamente dito, foi apresentada uma hipótese de avaliação para o grupo Bem Viver, sem a intervenção do programa. Desta maneira, realizou-se uma projeção para a utilização do grupo Bem Viver, a partir dos percentuais atingidos pelo grupo Recusa.