The Fallopian tube provides physiological support to both gametes and the embryo. Experimental approaches to improve the outcome of infertility treatment have included co-culture systems, whereby sperm, ova and/or embryos are cultured in a monolayer of Fallopian tube epithelial cells. This approach has been shown to be superior to other methods based on co-culture with other cell types. In addition, the role of gap junction intercellular communication (GJIC) in the regulation of cell proliferation and differentiation is becoming increasingly recognised as one of the major cellular functions. GJIC plays an important role in fertilisation as well as in the normal development of both the embryo and foetus. It is also involved in the sexual maturation of the adult and in the maintenance of health throughout life. As such, GJIC may be the key element in the understanding of cellular functions especially during the reproductive process. Therefore, the aims of this thesis were to examine some of the cellular and molecular aspects of the human Fallopian tube epithelium in relation to infertility. In the first part of this study, the gap junction proteins, connexins were examined by immunohistochemical and Western blotting techniques. Fallopian tubes were removed from women at different stages of the ovarian cycle. The results demonstrated the presence of gap junction proteins, connexins (cx) in the human Fallopian tube throughout the ovarian cycle. Fimbrial and ampullary regions were separated and subsequently processed for Western blotting analysis using a range of monoclonal or polyclonal antibodies directed against cx26, cx32 and cx43.The intensity of staining varied depending upon the hormonal status of the patients examined and appeared to be upregulated during the secretory stage as opposed to the proliferative phase of the ovarian cycle. Human Fallopian tube did not express cx32, regardless of the anatomical site examined. In all cases though, the expression of both cx26 and 43 appeared to be more prevalent in the ampullary region. The second part of this investigation focussed on transforming Fallopian tube epithelial cells into an immortal cell line. Normal cells cannot be sustained indefinitely, as cell degeneration occurs with continuous passaging. In most IVF treatments, sperm and ova are allowed to fertilise in a conventional medium but there is the potential that the Fallopian tube epithelial cells may be employed in IVF treatments in the future. Therefore, current techniques used to isolate and culture epithelial cells from the Fallopian tube were evaluated, in order to assess the best method that provides an optimal yield of Fallopian tube epithelial cells, both qualitatively and quantitatively. The cells were isolated using (i) a mechanical technique, whereby the mucosal layer of the tube were minced finely using a pair of scissors and (ii) an enzymatic method which consisted of the enzymes, trypsin and pancreatin. The results showed that enzymatic isolation provided a large number of cells, but there was a significant detrimental effect on cell survival and their secretory status. In contrast, cells isolated by the mechanical method were fewer in number than those obtained by the enzymatic method but cell survival and secretory status were relatively better. Regardless, of the isolation technique employed, the Fallopian tube epithelial cells had a limited lifespan in culture. Bearing this in mind and coupled with the realisation that the interactions between the tubal epithelial cells and gametes/embryos cannot adequately be studied in vivo, the next part of the study investigated ways of establishing an immortalised human Fallopian tube cell line. Such a cell line would be able to survive in culture indefinitely and therefore could be a potential model, which could mimic the in vivo milieu more closely. In order to obtain a 'bank' of Fallopian tube epithelial cells, cells isolated by either the mechanical or enzymatic method were cultured in growth medium for 24-48 hours prior to being transfected with a transforming gene, the SV40 Large T-antigen. Various techniques were used to transfect the epithelial cells in culture; these included well-established methods such as DNA co-precipitation as well as more advanced methods using cationic liposomes. The results from this study demonstrated that human Fallopian tube epithelial cells can be transiently transfected and could survive for over 102 days in culture, whilst still retaining many of the morphological characteristics of the original epithelial cell type. By examining both the cellular and molecular aspects of the tube as described above, it was anticipated that further insight into the complex nature of this structure would be achieved. Furthermore, it was hoped to improve the current understanding and knowledge of the Fallopian tube's functions, thereby facilitating a greater awareness of the issues important in establishing and maintaining an in vitro model of the Fallopian tube.

Neonates born to mothers with an active gonorrhoea infection can develop serious sight threatening eye infections. The causative agent, Neisseria gonorrhoeae, is passed to the neonate during birth, and infects the eyes of the neonate. The condition, ophthalmia neonatorum, develops 0-14 days after birth and initially presents as a painful inflammation of the eye with yellowish purulent discharge from one or both eyes. One preventative action used by some countries is the use of an ophthalmic prophylaxis which is usually an ophthalmic ointment which contains an antibiotic. This its self can cause chemical conjunctivitis. This study evaluates the potential to use a fatty acid or fatty acid derivative as the active antimicrobial agent in an ophthalmic prophylaxis. A panel of thirty-seven initial candidates were screened for anti-gonococcal properties. Seven of this panel were selected and tested against for ocular irritation potential using in vitro models and anti-gonococcal properties tested further in simulated tear fluid. Finally a single candidate, monocaprin, was selected as the main drug candidate. Ophthalmic formulations of liquid and semi-solid dosage forms were made and evaluated. Liquid dosage forms performed the best in in vitro tested and were further evaluated in cell culture and explanted models. The cell culture model suggested that monocaprin could be used to prevent infection 90 minutes after the cell were inoculated with the bacteria. An explanted corneal infection model was used to assess the potential formulations. It was shown that the anti-gonococcal properties of the drug candidate were inhibited on the ocular surface but this this could be countered by increasing the amount of monocaprin in the formulation. The formulations containing 0.188 % and 0.25 % (w/w) monocaprin were in some cases able to totally clear inoculations of higher cell numbers on the surface of the eye. Passage on agar plates containing monocaprin showed that increasing resistance due to genomic mutation is not likely and that existing mechanisms of fatty acid resistance did not give cross-resistance to monocaprin. However, duplicate samples passaged on monocaprin both acquired identical mutations in the dksA gene which may confer a small decrease in susceptibility. Also, work done on the processing of natural sources of fatty acids showed that treatment of coconut oil by use of a purified lipase or a lipase secreting yeast produced powerful anti-gonococcal substances. This could has the potential to be used in developing nations treat gonococcal and other bacterial infection. Overall, the work in thesis demonstrates that there is potential in the use a fatty acid or fatty acid derivative, most likely monocaprin, to be used as the active antimicrobial agent in an ophthalmic prophylaxis but more evaluation in terms of in vivo testing is required to demonstrate that the higher levels of monocaprin do not cause irritation to the eye.

Adoptive Cell Therapy of cancer using T cells is entering mainstream practice after years as a research method. Central to the efficacy of this “living therapy” is the function of the T cells transferred. T-cells, like other primary tissue cells, undergo differentiation and death. Clinical and preclinical data shows that lesser differentiated, less glycolytic, and more proliferative-capable cells used for the adoptive transfer yield superior tumor responses. This introductory section will describe discoveries which elucidate and control T-cell differentiation and function for the improvement of adoptive cell therapy. Namely by use of inhibition of the PI3k/AKT pathways, and through the discovery of a dual function of death and differentiation by the canonical death receptor Fas, which can be parsed apart with a mutation from valine to cysteine at the 194 position (C194V), differentiation can be withheld while leaving cell proliferation unhindered during T-cell stimulation and expansion. The data also reveals that the differentiation signal caused by extracellular Fas ligation passes through AKT, revealing both Fas and AKT as points of intervention for targeting differentiation along the same pathway. From further investigation, this introduction will describe the effect of AKT inhibition on T-cell differentiation on a transciptional and metabolic level. The data reveals AKT inhibition promoted FOXO1 intranuclear organization, which creates a more naive-like phenotype to the cells, and lower glycolytic status, another phenotype associated with persistent and long-lived cells. Furthermore, this control of AKT and Fas in T-cells yields benefits in several modalities of pre-clinical models of adoptive T-cell immunotherapy of cancer, in both use of a chimeric antigen receptor (CAR) and with use of Tumor Infiltrating Lymphocytes (TIL). Finally, the real-world applicability of the finding including the use of AKT inhibition in current approved Adoptive T-cell immunotherapies will be discussed.

Orientadores : Belmiro Gonçalves Pereira, Mary Angela Parpinelli / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-11-09T17:39:10Z (GMT). No. of bitstreams: 1 Ferreira_CarlosEduardo_M.pdf: 181051 bytes, checksum: acf346800333d34fb2e050b696c4e60e (MD5) Previous issue date: 2002 / Resumo: O objetivo deste estudo foi avaliar um novo protocolo de assistência prénatal de baixo risco implantado no Hospital das Clínicas da UNICAMP em julho de 1996 em relação ao diagnóstico de estados patológicos (anemia, infecção urinária, sífilis, HIV, hepatite B, toxoplasmose, rubéola, vulvovaginite e diabetes gestacional). Foi realizado um estudo de coorte, retrospectivo, operacional, tipo ensaio clínico com abordagem antes e depois da introdução do novo protocolo. Foram estudadas 250 gestantes deste protocolo e 250 gestantes do protocolo anterior. Os critérios de inclusão foram: gestantes que ingressaram no pré-natal até no máximo com 28 semanas de gestação e realizaram pelo menos quatro consultas. As análises dos dados, para algumas variáveis, foram realizadas pelo teste exato de Fisher e regressão logística. Para outras variáveis, as análises foram apenas descritivas. O nível de significância preestabelecido foi de 5%. Concluímos que o novo protocolo era composto de pacientes mais jovens, apresentou 10,8% e 29% de anemia, respectivamente no primeiro e terceiro trimestres de gestação, permitiu maior número de diagnóstico de infecção urinária e menor de sífilis em relação ao antigo protocolo. O novo protocolo identificou 45% e 25% de gestantes suscetíveis à toxoplasmose e rubéola, respectivamente. Em relação ao diabetes gestacional, o novo protocolo diagnosticou 17,4% de glicemia de jejum alterada. Vinte e seis por cento das gestantes do novo protocolo apresentaram vulvovaginite confirmada por bacterioscopia de secreção vaginal no primeiro trimestre e 18% no terceiro trimestre / Abstract: The objective of this study was to evaluate a new protocol for pre-natal care of women with low risk pregnancies. The new protocol, which was used from July 1996 on in the general hospital of the State University of Campinas (UNICAMP), served to diagnose the following pathologic conditions: anemia, urinary infection, syphilis, HIV, hepatitis B, toxoplasmosis, rubella, vaginitis and gestational diabetes. The study was of a retrospective, operational, cohort nature, involving a clinical trial which assessed conditions before and after the introduction of the new protocol. 500 patients were studied, of which 250 had been evaluated with the new protocol and 250 with the protocol that had been in use previously. The criteria for inclusion in the study were that the patients should have started pre natal care until the 28th gestational week and have had at least four medical evaluations. Data were analyzed with the help of Epi-Info statistical package, the Fisher¿s exact test or logistic regression being used for some variables, other analyses being merely descriptive. The level of significance defined was 5% (a=5%). The conclusion was that, in comparison with the old protocol, the new one allowed identification of the same number of cases of anemia, of a greater number of cases of urinary infection and of a lower number of cases of syphilis. The new protocol also allowed identification of cases of toxoplasmosis and rubella in, respectively, 45% and 25% of the patients. As to gestational diabetes, the new protocol allowed the identification of 17,4% cases of modified glycemy in patients under fasting. Twenty six percent of the pregnant women in the new protocol had vaginitis confirmed by vaginal smear at the first trimester and 18% at the third one / Mestrado / Tocoginecologia / Mestre em Tocoginecologia

Orientador: Maria Terezinha Serrão Peraçoli / Resumo: A pré-eclâmpsia é uma complicação grave da gestação, caracterizada por hipertensão e proteinúria. Os sintomas desta síndrome aparecem durante a segunda metade da gestação. A literatura sugere que a pré-eclâmpsia resulta de uma resposta inflamatória excessiva à gestação. O objetivo deste trabalho foi avaliar a atividade de monócitos de sangue periférico através da produção de citocinas, atividade fungicida e liberação de peróxido de hidrogênio por essas células em gestantes normais e com pré-eclâmpsia. Foram estudadas 25 mulheres não-grávidas e normotensas e 50 gestantes sendo 20 normotensas e 30 com pré-eclâmpsia. Monócitos foram cultivados com ou sem 10 ug/mL de lipopolissacáride (LPS) durante 24 h a 37oC e os níveis de fator de necrose tumoral alfa (TNF-a), de Interleucina-10 (IL -10) e de fator transformador do crescimento beta (TGF-b1) foram determinados no sobrenadante de cultura por ensaio imunoenzimático. A atividade fungicida foi avaliada pela recuperação de células viáveis de Candida albicans após plaqueamento e contagem de unidades formadoras de colônias em meio ágar Sabouraud. A liberação de peróxido de hidrogênio (H2O2) foi determinada antes e após estímulo dos monócitos com phorbol 12-myristate 13-acetate (PMA). Os resultados mostraram que os níveis endógenos de TNF-a, IL -10 e de TGF-b1 produzidos por monócitos, cultivados sem estímulo, foram significativamente mais elevados tanto em gestantes normais como nas com pré-eclâmpsia. Resultados semelhantes foram observados com relação à produção de IL-10 e TGF-b1 por monócitos após estímulo com LPS. A liberação de H2O2 após estímulo com PMA foi mais elevada em gestantes normais do que nas com pré-eclâmpsia. Além disso, monócitos de gestantes com... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Pre-eclampsia is a severe complication of pregnancy, characterized by hypertension and proteinuria. The symptoms of this syndrome appear during the second half of the pregnancy. The literature suggests that pre-eclampsia results from an excessive maternal inflammatory response to pregnancy. The objective of this study was to evaluate the activity of peripheral blood monocytes by cytokine production, fungicidal activity and hydrogen peroxide liberation in normotensive pregnant women and with pre-eclampsia. The study included 25 normotensive non-pregnant and 50 pregnant women being 20 normotensive and 30 with preeclampsia. Monocytes from these women were cultivated with or without 10 ug/mL lipopolysaccharide (LPS) during 24h at 37oC, and the levels of tumor necrosis factor-alpha (TNF-a), Interleukin-10 (IL -10) and transforming growth factor beta 1 (TGF-b1) were determined in culture supernatants by enzyme immunoassay. The fungicidal activity was determined by the recovery of viable Candida albicans after co-culture of yeasts with monocyte and plating in Sabouraud medium with enumeration of colony forming units. Hydrogen peroxide (H2O2) liberation was determined before and after monocyte stimulation with phorbol 12-myristate 13- acetate (PMA). The results showed that endogenous levels of TNF-a, IL-10 and TGF-b1 detected in supernatants of monocytes cultivated without stimulus were significantly higher in both normal and pre-eclampsia pregnant patients than in non-pregnant women. Similar results were observed in relation to IL -10 and TGF- b1 production after monocyte stimulation with LPS. Monocytes from normotensive pregnant women released higher levels of H2O2 after PMA stimulation than cells from patients with pre-eclampsia. Besides monocytes from women with preeclampsia produced lower levels of TNF-a suggesting an impairment in the... (Complete abstract click electronic address below) / Mestre

Dentro de um enfoque vygotskyano, o presente estudo investigou a passagem da heterorregulação para a auto-regulação nas interações de mães/pais-criança. Mais especificamente, procurou identificar os primórdios do aparecimento da autonomia na criança e as situações interativas que a fazem manifestar-se. Participaram do estudo 12 famílias, compostas por pai, mãe e criança. As crianças eram de ambos os sexos e tinham 2 anos de idade. Elas permaneceram dez minutos com cada um dos pais, explorando objetos não-familiares em sessões de observação em que as díades foram filmadas. Os dados foram analisados de forma quantitativa, utilizando-se a estatística descritiva e inferencial; e qualitativa, através da microanálise. Os resultados não apresentaram diferenças significativas em relação a ambos, os comportamentos indicativos de hétero e auto-regulação dos pais e das mães e os comportamentos das crianças na interação com os pais e com as mães. Também mostraram que aos 2 anos de idade já é possível observar os primórdios da passagem da hétero para a auto-regulação, sugerindo relação entre tipo de interação - heterorregulação indireta -, desenvolvimento cognitivo da criança e a passagem da hétero para a auto-regulação. Discute-se os resultados, mostrando que o processo de interação é moldado nem pelas características dos pais nem pelas características da criança, mas pela criação de um espaço intersubjetivo na interação adulto-criança que, por sua vez, reflete as práticas do grupo cultural, representado pela classe social.

Syftet med den här studien är att synliggöra svårigheter som uppkommer vid undervisning inom pre-algebra. Pre-algebran och algebrans roll i skolan är viktig därför att den medför att elever lär sig se problem ur olika synvinklar. Detta ger elever möjligheten att utveckla deras kognitiva tankemönster. Studien tar upp flera kritiska aspekter samt variationsmönster för att belysa svårigheter i undervisningen inom pre-algebra. Vidare beskrivs hur läraren kan gå till väga för att motverka vissa svårigheter. Resultatet visar på viktiga delar som berör inlärningen av pre-algebra. Det handlar bland annat om likhetstecknets betydelse, lärarens ämneskunskaper, ordanvändning, rutiner och relationella tal. Slutligen visar forskningen att inte enbart elevers kunskaper som är bristfälliga, utan även lärarens kunskap kan vara otillräcklig.

Duchenne muscular dystrophy (DMD) is a severe, progressive muscle wasting disorder that affects 1 in 3600 male births. It is caused by genetic mutations in the dystrophin gene. This study investigated several aspects of the neuromuscular disorder within a population of Brazilian DMD boys and their families. This study's framework was laid out within the prism of an interacting cycle of genetic factors, cognitive functioning, and psychosocial aspects that underlie the neuromuscular disorder. It focuses on DMD's aetiology, history and previous research on genetic, cognitive and psychosocial aspects. Mixed methods were adopted to allow for a more encompassing view of the neuromuscular disorder: cognitive tests, an emotion recognition battery, genetic analyses, well-being questionnaires, and interviews were applied. Correspondent, quantitative and qualitative data analysis was carried out. The findings of 32 DMD patients (mean age 10.4 years, SD= 2 years) and 31 control subjects (mean age 9.4 years, SD= 3 years) revealed severe cognitive dysfunctioning in all assessed cognitive domains in the DMD population, as well as in the ability of emotion recognition. In the DMD group, it could be shown that poor executive functioning stood in a positive correlation with a poor ability of emotion recognition. The DMD patients' cognitive phenotypes were correlated with the genetic mutations in their dystrophin gene, but no relationship between the patients' genotype and cognitive phenotype could be confirmed. These results were contrary to previous research, which suggested that specific mutations in the dystrophin gene cause cognitive impairment. The DMD group scored poorly on the emotion recognition task, which is also a characteristic of autism spectrum disorder. However, when diagnosing for autistic characteristics through means of an interview, only a few similarities between the two disorders could be found. In order to assess the psychosocial components that come along with the disorder, well-being questionnaires were supplied. Interestingly, DMD boys scored higher on well-being than the boys in the control group. Moreover, 30 of the DMD caregivers (mean age app. 31 years) also revealed high levels of well-being, which correlated positively with the well-being of their sons, suggesting high levels of resilience. Given the participants' socio-economic hardship and the lack of governmental help, it was concluded that participants showed an incredible level of resilience that most likely resided within their faith, which nearly all of them stated to be the reason for their strength to strive. The relevant and new information about cognitive, genetic and social aspects of DMD uncovered in this study will pave the way for further (and much needed) studies into psychosocial aspects of the disorder.

abstract: Recidivism occurs when an individual is released from prison and then, through a violation of parole or a new offense, ends up back in prison. Durose and colleagues (2014) cite that 55.4% of individuals go back to prison after a five-year post release. Considerable attention has been focused on reducing the cycle of these individuals going back to prison. One of the attempts to remedy this issue is through offering pre-release programs for prison inmates. These programs seek to provide individuals skills that will reduce their likelihood of reoffending. But existing research shows that the effectiveness of these programs is limited. Moreover, few attempts have been made to look at differences between individual’s dosage of program participation. This thesis aims to determine if participation in a pre-release program reduces recidivism. Using data from the state’s Department of Corrections, there is a comparison of previously imprisoned individuals who participated in a pre-release employment program and those who declined participation, to understand if participation influences recidivism. Additionally, dosage of the center will be analyzed to determine whether length of program participation influenced recidivism. Participating in the program and a longer dose of the program should allow more time for the individual to learn the material and fully absorb what the program is offering such as skills and training. The results show that participation in the pre-release employment program did not significantly affect recidivism as individuals who went through the program were no less likely to be reincarcerated. In addition, a longer dosage of the program did not significantly influence recidivism among those who went through the program. / Dissertation/Thesis / Masters Thesis Criminology and Criminal Justice 2019

Nearly 70% of preschool children in the United States are enrolled in child care facilities. This means that they eat many meals away from their homes. Despite government support for childhood nutrition through the Child and Adult Care Food Program (CACFP), research that measures the nutritional value of meals served in child care facilities has been lacking. The purpose of this quantitative study was to determine if there are differences in the calories and nutritional value of lunch meals offered to preschool children in facilities that participate in CACFP and in facilities that do not participate in CACFP. Ajzen's theory of planned behavior formed the theoretical foundation for this study. Two research questions addressed the nutrient and caloric content of lunches served in CACFP and non-CACFP facilities. An ex post facto quasi experimental design was used to compare 598 meals from existing monthly menus from a random sample of 30 child care facilities located in a state in the Southwestern United States. Using a MANOVA test, significantly greater amounts of proteins, fats, and calories were found in meals served by non-CACFP facilities. A comparison of actual menu items suggested that greater numbers of fatty foods were present in menus served at non-CACFP facilities. These results support literature that found childhood illnesses, like obesity and malnutrition, may stem from high-calorie meals that lack adequate nutrients. This study may contribute to positive social change by supporting nutrition oversight, such as that provided by the CACFP program; encouraging tighter state and local nutritional guidelines in child care; and focusing attention on the importance of everyday nutrition for all children attending child care facilities.