"I'm sure you'll figure out that since I'm telling you this story, I didn't die when it happened… But someone did." Thus begins the story of Matt Eyering and his journey from Staten Island to the small New Mexican town of Socorro. He goes for one reason: to exact revenge on his estranged mother. But all sorts of odd characters live in this forgotten place along the Rio Grande, and none of them intends to let Matt pursue his goals smoothly. From revolutionaries to drug lords, romance to addiction, revenge to life threatening illness, what begins as a quest for vindication explodes into an adventure through a gauntlet of small-town politics, class discrimination, racial tension, organized crime, and self-discovery. It is a touching and serious, yet humorous exploration of the human condition, and of life in a place most of America conveniently ignores.

Alcohol use and abuse has many harmful effects, especially to children exposed prenatally, including fetal alcohol spectrum disorders (FASDs). The disabilities due to fetal alcohol exposure continue throughout life and cause major financial burdens to society. The molecular mechanisms underlying FASDs are not well understood. We have taken a genetic approach to characterize ethanol’s effect on changing a discrete cell fate decision during embryogenesis in the nematode, Caenorhabditis elegans (C. elegans). Our preliminary data suggest that ethanol can affect the development of AWC neurons, a pair of olfactory neurons in C. elegans. We suggest that lipids can protect AWC neurons from ethanol’s effects. Importantly, we show that altering the metabolism of triacylglycerols (TAGs) can rescue this cell fate change in behavioral assays. By identifying molecular causes of fetal alcohol damage in humans we hope to be able to develop a greater understanding of how to prevent these detrimental effects.

Fetal alcohol spectrum disorder is the leading cause of mental retardation in the western world. It is associated with learning and sensory deficits. Some of these deficits are a result of faulty neuronal plasticity. Previously our lab has used ferrets to demonstrate that alcohol exposure during the third trimester of human gestation results in impaired ocular dominance plasticity (ODP). Here we have transferred this model to mice. Mice, treated with 5 mg/kg of ethanol on postnatal days 5, 7 and 9, exhibit a lack of ODP plasticity after 10 days of monocular deprivation (MD) during the critical period of visual cortex plasticity, as seen by optical imaging of intrinsic signals. This deficit in ODP was rescued by treatment with a phosphodiesterase type 1 inhibitor (PDEi1), vinpocetine. This rescue did not occur after treatment with a PDEi4 (rolipram) or a PDEi5 (vardenafil) inhibitor alone. Interestingly when these drugs were given concurrently, ODP was rescued. To further explore the effect of early alcohol exposure on ODP, we used Visually Evoked Potentials to examine the potentiation and depression components of ODP. Ethanol exposed and saline control animals were MD for 5, 7 or 10 days during the critical period of the visual cortex. Here we saw that although saline animals exhibited a normal depression of contralateral eye responses and a potentiation of ipsilateral eye responses, ethanol animals exhibited only a depression of contralateral eye responses. Additional ethanol animals were then MD for 3 days to test for changes in the on-set of contralateral eye depression. Yet, these animals exhibited normal contralateral eye response changes. In conclusion early ethanol exposure disrupts only the potentiation of the ipsilateral eye inputs, while leaving the contralateral eye response depression in tact. This model provides a new approach to studying ODP after early alcohol exposure, opening the door for studies using transgenic animals to further elucidate the mechanisms behind these alcohol induced deficits.

Purpose: To compare the determinants and distribution of fetal and early neonatal deaths in the Commonwealth of Virginia. Background: Much attention is devoted to reducing the infant mortality rate which was declining up until 2002. The recent rise was parsed and found to stem from an increase in deaths during the early neonatal period. Fetal deaths are not well understood and are not routinely included when evaluating infant mortality. Methods: Using data collected from 2001 to 2006 fetal death and linked infant birth and death certificates by the Virginia Department of Health, crude mortality rates and leading causes of death were calculated for fetal and early neonatal mortality. Rates were calculated for each period of death by locality and mapped to determine if the distribution differed. Logistic regression was also used to evaluate sociodemographic and pregnancy risk factors and chi-square analyses were used to determine if the determinants varied significantly by timing of death outcome. Results: During the study period, the fetal death rate was 5.4 per 1,000 fetal deaths plus live births, the early neonatal death rate was 2.5 deaths per 1,000 live births and perinatal mortality rate was 7.9 deaths per 1,000 fetal deaths plus live births. Trends over time, gestational age specific mortality, geographic distribution, cause of death and many determinants were comparable between both death periods. Extremely low birth weight was the most significant risk factor for early neonatal death (OR = 1747.06). Congenital anomalies of the child were the leading predictor of fetal death (OR = 26.24, 95% CI: 19.62, 35.10) and second highest for early neonatal death (OR = 52.26, 95% CI: 35.21, 77.56). Conclusions: Because of the similarities in geographic distribution, sociodemographic factors, pregnancy risk factors and causes of death, analyzing neonatal and infant mortality rates in isolation from fetal deaths does not accurately depict the burden of adverse pregnancy outcomes.

In this thesis I evaluate the impact of an experiment with free sales of strong beer in two Swedish counties that took place in the 1960s. I do this by studying adult earnings of persons in utero during the experiment. My data includes date and place of birth and allows me to evaluate the impact of the experiment using a difference-in differences methodology, comparing earnings across cohorts and counties. Since the availability of alcohol increased most heavily for persons under the age of 21, and male fetuses are less physiologically robust than female fetuses, I choose to study persons born by mothers younger than 21 separately and also estimate the impact of the experiment separately for men and women. I find that persons born by mothers under the age of 21 during the experiment have lower average earnings than persons born before the experiment, and that the impact is larger on men. My results indicate that the experiment has led to adverse effects on adult earnings, probably caused by the prenatal alcohol exposure’s negative impact on fetal development. This means that alcohol consumption have long-term consequences that represent large costs to society. Since these costs are generally disregarded when evaluating the cost of alcohol consumption, society’s cost of alcohol is probably higher than usually estimated.

Despite many attempts to improve estimations based on ultrasound measures and volumes, the accurate prediction of birthweight hasremained elusive, particularly in relation to macrosomia.Now finally, after 7 years of dedicated research, we have managed to develop a rapid and precise, magnetic resonance imaging-basedtechnique, capable of accurately predicting birthweight, regardless of whether the estimations are performed, hours, days or even severalweeks prior to the actual birth.We believe that, once the already demonstrated advantages of the technique are confirmed in prospective, multi-centre studies, itwill become readily accessible for use in clinical practice and prove invaluable to clinicians as a reliable adjunct in many obstetricaldecision-making processes. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

Identifica la correlación entre el área de la gelatina de Wharton en un corte transversal y el peso fetal por encima del percentil 90 evaluados por ultrasonografía en gestantes a término del Servicio de Diagnóstico Médico – Lima entre los meses de octubre - noviembre 2018. El estudio es de tipo observacional, prospectivo y de corte transversal, de nivel relacional, con diseño correlacional. El tamaño de la muestra fue de 96 ultrasonografías en gestantes a término atendidas en el Servicio de Diagnóstico Médico de octubre a noviembre del 2018. Para analizar la correlación entre las variables se utilizó la prueba Rho de Spearman y la prueba no paramétrica de Fisher. El área promedio de la gelatina de Wharton fue de 236.4 mm2 y el 84.4% del área de la gelatina de Wharton se encuentra por encima del percentil 95 y el 15.6% por debajo del percentil 95%. El promedio del peso fetal según método Hadlock IV fue de 3794.2 gr, encontrándose el 90.6% del peso fetal se encuentra por encima del percentil 90. Asimismo se evidencia que existe una relación directa, moderada y muy significativa entre el área de la gelatina de Wharton medida por ultrasonografía y el ponderado fetal por Hadlock IV, obteniéndose un coeficiente de correlación de Rho de Spearman = 0.651, con un ρ = 0.000. Existe relación significativa entre el área de la Gelatina de Wharton por encima del percentil 95 con la presencia de macrosomía según peso real al nacer (p=0.000). Se concluye que existe correlación significativa entre el área de la Gelatina de Wharton en un corte transversal y el peso fetal por encima del percentil 90 evaluados por ultrasonografía en gestantes a término del Servicio de Diagnóstico Médico – Lima entre los meses de octubre - noviembre 2018. / Tesis

Com o objetivo de avaliar a influência do puerpério e da retenção dos anexos fetais no proteinograma de fêmeas bovinas da raça Holandesa, foram colhidas 291 amostras de sangue de vacas nos primeiros 90 dias após o parto. Para estudo do puerpério fisiológico, foram colhidas amostras de sangue de 162 fêmeas bovinas clinicamente sadias, divididas em 9 grupos experimentais, 0 -| 1º dia; 1º -| 2º dia; 2º -| 4º dia; 4º -| 6º dia; 6º -| 8º dia; 8º -| 15º dia; 15º -| 30º dia; 30º -| 60º dia; e, 60º -| 90º dia pós-parto. Para avaliar a influência da retenção dos anexos fetais, foram colhidas 129 amostras de soro sanguíneo de vacas da raça Holandesa que retiveram os anexos fetais, divididas em 8 grupos experimentais, 1º -| 2º dia; 2º -| 4º dia; 4º -| 6º dia; 6º -| 8º dia; 8º -| 15º dia; 15º -| 30º dia; 30º -| 60º dia; e, 60º -| 90º dia pós-parto. Os teores séricos de proteínas totais foram determinados pela técnica de Biureto e o fracionamento das proteínas foi realizado por eletroforese em fita de acetato de celulose e eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Os resultados permitem concluir que ocorreu um aumento significativo das proteínas de fase aguda tanto nos animais com puerpério fisiológico quanto nos animais com retenção dos anexos fetais. As proteínas haptoglobina, ceruloplasmina e glicoproteína ácida responderam ao estímulo do parto e à contaminação bacteriana pós-parto nos animais com puerpério fisiológico; e nos animais com retenção dos anexos fetais, responderam de forma significativa ao estímulo da contaminação bacteriana pós-parto e da retenção dos anexos fetais prolongando o tempo de resposta das proteínas. A glicoproteína ácida representou mais evidentemente o quadro de inflamação aguda induzida pela retenção dos anexos fetais. Nos animais com retenção dos anexos fetais, a proteína albumina comportou-se como proteína de fase aguda negativa, provavelmente pela interferência de vários fatores, como distúrbios hepáticos ou metabólicos; desvio de sua síntese hepática para a produção de outras proteínas; e inflamação, infecção e absorção de toxinas bacterianas. A técnica de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE) a 10%, com tampão de aplicação com agente redutor, não é indicada para o estudo do comportamento das proteínas albumina e imunoglobulinas. Pela técnica de eletroforese em fita de acetato de celulose demonstrou-se que o puerpério não alterou de forma significativa a concentração de albumina, mas alterou significativamente as frações alfa, beta e gamaglobulinas. / The aim of this study was to evaluate the influence of the puerperal period and retained fetal membranes on the proteinogram of Holstein cows. Two hundred ninety one blood samples were collected from cows through ninety days postpartum. In order to study the physiologic puerperium, blood samples from one hundred two clinically animals, divided in 9 experimental groups, following the chronogram, 0 -| 1º day; 1º -| 2º day; 2º -| 4º day; 4º -| 6º day; 6º -| 8º day; 8º -| 15º day; 15º -| 30º day; 30º -| 60º day; and, 60º -| 90º day postpartum were analyzed. To evaluate the fetal membranes retention influence, one hundred twenty nine blood samples from Holstein cows with this clinical condition, in moments1º -| 2º day; 2º -| 4º day; 4º -| 6º day; 6º -| 8º day; 8º -| 15º day; 15º -| 30º day; 30º -| 60º day; and, 60º -| 90º day postpartum were also analyzed and divided in 8 experimental groups. Total proteins concentration was determined by the Biuret method and protein electrophoresis were performed in acetate cellulose and in sodium dodecyl sulfate polyacrilamide gel (SDS-PAGE). The results showed a significant elevation in acute phase proteins in both conditions, physiologic puerperium and retained fetal membranes. Haptoglobin, ceruloplasmin and acid glycoprotein were increased in the partum stimuli and postpartum bacterial contamination in animals with physiologic puerperium. In the retained fetal membranes animals, those proteins significantly responded to the pos-partum bacterial contamination stimulli and retained fetal membranes, rising the proteins response time. The acid glycoprotein concentration evidenciated an acute inflammation induced by retained fetal membranes. In the animals with this condition, albumin behaved as negative acute phase protein, probably by several factors like hepatic or metabolic disturbance, deviation of the hepatic synthesis secondary to other proteins production, and bacterial toxins absortion, inflammation and infection. Sodium dodecyl sulfate poliacrilamide 10% gel (SDS-PAGE) electrophoresis with reducing sample aplication buffer, is not indicated for albumin and immunoglobulins behavior study. Acetate cellulose electrophoresis showed that physiologic puerperium did not influencied albumin concentration, but significantly did with alfa, beta and gammaglobulins.

O álcool é uma das substâncias psicoativas mais consumidas mundialmente e seu uso por mulheres em idade reprodutiva, em particular, tem representado grande preocupação por parte de especialistas e da sociedade em geral. Apesar dos efeitos adversos associados ao ato de ingerir bebidas alcoólicas durante a gestação ser bastante documentados e conhecidos, sabe-se que uma parcela de mulheres grávidas tem dificuldades em abandonar o hábito. O consumo excessivo de álcool durante a gravidez tem sido associado com a síndrome fetal pelo álcool (FAS), caracterizada por crianças com dificuldades comportamentais e de aprendizado. Entretanto, devido ao sentimento de culpa e medo de ações punitivas, mulheres raramente admitem terem utilizado álcool durante a gestação. Como resultado, uma série de marcadores biológicos tem sido estudada para se diagnosticar a exposição fetal ao etanol. Dentre os marcadores utilizados estão os ésteres etílicos de ácidos graxos (FAEE), que podem ser detectados em amostras de mecônio de recém-nascidos. No presente trabalho, um método analítico foi desenvolvido visando a detecção de oito FAEEs em amostras de mecônio e aplicada em amostras coletadas de recém-nascidos cujas mães admitiram ou não o uso de etanol durante a gestação. A microextração em fase sólida por Headspace (HS-SPME), uma técnica de preparação de amostras relativamente recente, foi utilizada para análise. Os FAEEs foram identificados e quantificados por cromatografia gasosa/espectrometria de massas (GC/MS), operado no modo de ionização química. Os correspondentes ésteres etílicos deuterados foram sintetizados e utilizados como padrão interno. Os limites de quantificação (LOQ) obtidos foram abaixo de 150 ng/g e limites de detecção (LOD) foram abaixo de 100ng/g para todos os analitos. O método mostrou boa linearidade na concentração estudada (LOQ-2000ng/g), com coeficiente (r2) melhor que 0.98. Os valores de precisão apresentaram coeficientes de variação menores que 15% para todos os FAEEs estudados. Quando o método foi aplicado em amostras de mecônio, foi possível detectar níveis de alguns FAEEs de recém-nascidos não suspeitos a exposição fetal ao etanol. / Alcohol is the main psychoactive drug consumed worldwide and its increasing use by young women has been a great problem point out by specialists in the subject. Although the adverse effects associated to the habit of drinking alcoholic beverages during gestation being very much documented, it is known that a considerable number of pregnant women have difficulties to abandon the habit. Excessive alcohol use during pregnancy has been associated with Fetal Alcohol Syndrome (FAS) characterized by children with cognitive and behavioral disorders. However, because of denial, embarrassment and fear, maternal reports of gestational use of alcohol are often inaccurate. Consequently, a series of biomarkers have been studied to diagnose fetal exposure to alcohol. Recently, fatty acid ethyl esters (FAEE) have been studied as biomarkers found in meconium of neonates exposed in utero. In the present work, an analytical method was developed aiming the detection of eight FAEEs in meconium samples and applied to real specimens collected from newborns whose mothers admitted or not the use of alcohol during pregnancy. Headspace solid-phase microextraction (HS-SPME), a relatively recent sample preparation technique, was used for analysis. FAEEs were identified and quantified by gas chromatography/mass spectrometry (GC/MS), operated in chemical ionization mode. The corresponding deuterated ethyl esters were synthesized and used as internal standards. The lower limits of quantification (LOQ) obtained were below 150ng/g and limits of detection (LOD) were bellow 100ng/g for all analytes. The method showed good linearity in the range of concentration studied (LOQ-2000ng/g), with coefficient of linearity better than 0.98. The precision assay, given by the relative standard deviation (RSD) of the method was lower than 15% for all FAEEs studied. When the method was applied to real samples, it was possible to detect trace levels of some FAEEs from non-suspected ethanol exposed newborns.

Since 2011 the prenatal diagnosis field has undergone a revolution with the introduction of a noninvasive prenatal test (NIPT) for genetic diseases relying on analysis of fetal cell-free DNA present in maternal plasma. Although available in Brazil, we rely on outsourcing the technology developed abroad or the test itself. Therefore, our objective was to develop and implement a comprehensive NIPT using high-coverage targeted next-generation sequencing to: 1) estimate fetal fraction; 2) determine fetal sex; 3) detect trisomy; 4) detect monogenic disease. We developed a robust and accurate model (r2= 0.994, p-value < 2.2e-16) for fetal fraction estimation based on distribution of SNP minor allele fraction (MAF). We used Z-score for fetal sex determination (100% accuracy) and trisomy detection of chromosomes 21 (T21) and 18 (T18), achieving a sensitivity of 100% (95% CI: 63.06% - 100.00%) and a specificity of 98.53% (95% CI: 92.08% - 99.96%) for T21, and 40% (95% CI: 5.27% - 85.34%) and 98.59% (95% CI: 92.40% - 99.96%) for T18. For monogenic disease detection (skeletal dysplasia) we performed variant analysis, with 71% (5/7) of detection rate. To our knowledge, this is the first work to integrate all analysis in one single test, and to perform monogenic disease detection without using parental genotype. We showed in this work that it is possible to implement such techniques in our country, using available resources and/or engaging in collaboration with reference research groups abroad. It shows the potential of developing internal technologies, and applying it to other noninvasive fields, such as cancer and organ rejection diagnostic and monitoring / Desde 2011 a área de diagnóstico pré-natal sofreu uma revolução com a introdução de teste pré-natal não-invasivo (NIPT) de doenças genéticas, que se baseia na análise de DNA fetal livre de células presente no plasma materno. Apesar de estar disponível no Brasil, nós dependemos em terceirizar a tecnologia desenvolvida no exterior, ou o teste em si. Sendo assim, nosso objetivo foi desenvolver e implementar um teste NIPT abrangente utilizando sequenciamento de nova geração de alta cobertura targeted para: 1) estimar fração fetal; 2) determinar sexo fetal; 3) detectar trissomia; 4) detectar doença monogênica. Nós desenvolvemos um modelo robusto e preciso (r2= 0.994, p-value < 2.2e-16) para estimativa de fração fetal baseado na distribuição da fração alélica de SNPs. Nós utilizamos Z-score para determinação de sexo fetal (100% de precisão) e detecção de trissomia dos cromossomos 21 (T21) e 18 (T18), atingindo uma sensibilidade de 100% (95% IC: 63.06% - 100.00%) e especificidade de 98.53% (95% IC: 92.08% - 99.96%) para T21, e 40% (95% IC: 5.27% - 85.34%) e 98.59% (95% IC: 92.40% - 99.96%) para T18. Para detecção de doença monogênica (displasia esquelética) nós realizamos análise de variante, com uma taxa de detecção de 71% (5/7). Até onde sabemos, este é o primeiro trabalho a integrar todas as análises em um único teste, e a realizar detecção de doença monogênica sem utilizar genótipo parental. Nós mostramos neste trabalho que é possível implementar essas técnicas no nosso país, utilizando recursos disponíveis e/ou desenvolvendo colaborações com grupos referência internacionais. Isto demonstra o potencial de desenvolver tecnologias internas, e aplicá-las à outras áreas não-invasivas, como diagnóstico e monitoramento de câncer e rejeição de transplante