<p>Functioning as a “powerhouse”, the liver adapts to the metabolic needs of the body by maintaining a homeostatic balance. Prolactin receptor (PRLR) has been found to have a copious existence in the liver. Having established a well-defined role in both reproductive and endocrine systems, the role of this transmembrane protein in hepatocytes is yet to be elucidated. Due to its abundant nature, we hypothesized that PRLR is required for maintaining hepatic homeostasis and plays a role in liver diseases. To test this hypothesis, we defined two specific aims. The first was to explore whether PRLR loss-of-function affects liver structure and function in physiological conditions. The second was to determine whether PRLR is associated with liver pathology. We deleted the <i>Prlr</i> gene specifically in hepatocytes using a virus-based approach and evaluated liver function, transcriptome, and activities of downstream signaling molecules. Due to the absence of PRLR, we found that the urea cycle was disrupted, concomitant with excessive accumulation of urea in the blood; 133 genes exhibited differential expression, largely associated with hepatocyte structure, metabolism, and inflammation; and the activities of STAT3 and 5 were reduced. The results signify that PRLR indeed plays a homeostatic role in the liver. We also used <i>Prlr</i>^+/- mice to assess whether the loss of one allele of the <i>Prlr</i> gene alters maternal hepatic adaptations to pregnancy. As a result, in the pre-pregnancy state and during the first half of gestation, the expression of maternal hepatic PRLR protein was reduced approximately by half owing to <i>Prlr</i> insufficiency. However, during the second half of pregnancy, we observed compensatory upregulation of this molecule, leading to minimal interference in STAT 3 and 5 signaling and liver size. Contrary to a previous study in the breast and ovary, our results suggest that one allele of <i>Prlr</i> may be sufficient for the maternal liver to respond to this physiological stimulus (pregnancy). Furthermore, we examined the expression and activity of PRLR in fatty as well as cholestatic livers. Using a high fat diet, we induced non-alcoholic fatty liver disease (NAFLD). Strikingly and for the first time, we discovered that the short isoform of PRLR (PRLR-S) was completely inactivated in response to NAFLD, whereas the long isoform remained unchanged. This finding strongly suggests the involvement of PRLR-S in lipid metabolism. We also postulate that PRLR-L may be the major regulator of STAT signaling in the liver, consistent with other reports. Lastly, we induced extrahepatic cholestasis via bile duct ligation (BDL) in mice. As this liver disease progressed, the expression of both isoforms of PRLR generally declined and was surprisingly accompanied by increased STAT 3 and 5 activity. The data suggests that PRLR participates in this disease progression, with a disconnection between PRLR signaling and STAT proteins. Collectively, our preliminary studies suggest that PRLR signaling is required to maintain liver homeostasis and more prominently, is involved in liver diseases, especially NAFLD. These findings lay a foundation for our future studies.</p>

Indiana University-Purdue University Indianapolis (IUPUI) / Functioning as a “powerhouse”, the liver adapts to the metabolic needs of the body by maintaining a homeostatic balance. Prolactin receptor (PRLR) has been found to have a copious existence in the liver. Having established a well-defined role in both reproductive and endocrine systems, the role of this transmembrane protein in hepatocytes is yet to be elucidated. Due to its abundant nature, we hypothesized that PRLR is required for maintaining hepatic homeostasis and plays a role in liver diseases. To test this hypothesis, we defined two specific aims. The first was to explore whether PRLR loss-of-function affects liver structure and function in physiological conditions. The second was to determine whether PRLR is associated with liver pathology. We deleted the Prlr gene specifically in hepatocytes using a virus-based approach and evaluated liver function, transcriptome, and activities of downstream signaling molecules. Due to the absence of PRLR, we found that the urea cycle was disrupted, concomitant with excessive accumulation of urea in the blood; 133 genes exhibited differential expression, largely associated with hepatocyte structure, metabolism, and inflammation; and the activities of STAT3 and 5 were reduced. The results signify that PRLR indeed plays a homeostatic role in the liver. We also used Prlr+/- mice to assess whether the loss of one allele of the Prlr gene alters maternal hepatic adaptations to pregnancy. As a result, in the pre-pregnancy state and during the first half of gestation, the expression of maternal hepatic PRLR protein was reduced approximately by half owing to Prlr insufficiency. However, during the second half of pregnancy, we observed compensatory upregulation of this molecule, leading to minimal interference in STAT 3 and 5 signaling and liver size. Contrary to a previous study in the breast and ovary, our results suggest that one allele of Prlr may be sufficient for the maternal liver to respond to this physiological stimulus (pregnancy). Furthermore, we examined the expression and activity of PRLR in fatty as well as cholestatic livers. Using a high fat diet, we induced non-alcoholic fatty liver disease (NAFLD). Strikingly and for the first time, we discovered that the short isoform of PRLR (PRLR-S) was completely inactivated in response to NAFLD, whereas the long isoform remained unchanged. This finding strongly suggests the involvement of PRLR-S in lipid metabolism. We also postulate that PRLR-L may be the major regulator of STAT signaling in the liver, consistent with other reports. Lastly, we induced extrahepatic cholestasis via bile duct ligation (BDL) in mice. As this liver disease progressed, the expression of both isoforms of PRLR generally declined and was surprisingly accompanied by increased STAT 3 and 5 activity. The data suggests that PRLR participates in this disease progression, with a disconnection between PRLR signaling and STAT proteins. Collectively, our preliminary studies suggest that PRLR signaling is required to maintain liver homeostasis and more prominently, is involved in liver diseases, especially NAFLD. These findings lay a foundation for our future studies.

No description available.

This diploma thesis deals with preparation and characterization of alcohol-free beers with different flavours. Five flavoured and four pure, commercially available beers were tested. Three alcohol-free beers were then selected and used for preparation of flavoured beers. Ingredients used for production of flavour were honey, raspberry, ginger and cranberry. Analysis was focused mainly on substances of phenolic nature. Flavoured beers were also tested in sensory analysis. Ingredients used for flavouring of beers and production of both alcoholic and alcohol-free beers are described in theoretical part of the thesis. Analysis of polyphenols, flavonoids, technological characteristics and anthocyanins was performed using spectrophotometric method. Phenolic substances were identified and quantified by HPLC/PDA. Ascorbic acid was analysed by HPLC-NH2/UV. Results proved an increase of polyphenols, ascorbic acid and anthocyanins during flavouring of alcohol-free beers in dependence of used ingredients. Sensory analysis showed that as the best rated was the Staropramen beer with raspberry flavour. On the contrary, flavoured Bernard beers were poorly rated.

La bronchopneumopathie chronique obstructive (BPCO) est une des maladies chroniques les plus fréquentes et constitue une des premières causes de mortalité dans le monde avec un impact sociétal majeur et des couts de santé considérables. La BPCO est aujourd’hui considérée comme une maladie multi-systémique dont le pronostic est lié en grande partie à ses comorbidités et à la survenue d’exacerbations. Les exacerbations qui vont ponctuer l’évolution de la BPCO précipitent le déclin de la fonction respiratoire et favorisent la décompensation des comorbidités et la survenue d’événements cardiovasculaires comme des infarctus du myocarde ou des accidents vasculaires cérébraux. La prise en charge moderne de la BPCO est basée sur la mise en place d’un soin intégré incluant la prise en charge des comorbidités et une meilleure détection et gestion des exacerbations.Dans ce travail de thèse nous abordons l’atteinte hépatique (stéatopathie hépatique non alcoolique - NAFLD) dans la BPCO comme une comorbidité sous-estimée (publication 1) alors qu’elle a probablement des implications pronostiques importantes (publication 2). La BPCO s'accompagne également de symptômes non respiratoires tel que des douleurs, dont les variations et localisations sont peu connues pendant et après l'exacerbation (publication 3). Le traitement de ces douleurs par des opiacés pourrait avoir un effet spécifiquement délétère dans cette population (publication 5). Les enjeux de réforme du système de santé avec une optimisation cout-efficacité incitent à développer et valider de nouvelles méthodes de prise en charge ambulatoire des exacerbations (publication 6).Dans une première partie de la thèse nous explorons les liens épidémiologiques entre BPCO et NAFLD. Nous explorerons également les conséquences d’une telle association sur le devenir cardiovasculaire des patients à moyen terme. Dans une deuxième partie, nous cherchons à définir les caractéristiques des douleurs avant et après exacerbation au cours de la BPCO, le lien entre douleur, anxiété et dépression chez ces patients et la sécurité d'emploi des morphiniques dans cette population fragile. Dans une dernière partie, nous aborderons la stratification du risque lié à une exacerbation de BPCO, et la possibilité d’une prise en charge ambulatoire extrahospitalière pour les exacerbations de sévérité modérée. / Chronic Obstructive Pulmonary Disease (COPD) is one of the most common chronic diseases and is one of the leading causes of death in the world with major societal impact and considerable health costs. COPD is now considered a multi-systemic disease whose prognosis is largely related to its comorbidities and the occurrence of exacerbations. The exacerbations that punctuate the evolution of COPD precipitate the decline of respiratory function and promote the decompensation of comorbidities and the occurrence of cardiovascular events such as myocardial infarction or cerebrovascular accidents. The modern management of COPD is based on the implementation of integrated care including the management of comorbidities and better detection and management of exacerbations.In this thesis we address liver injury (non-alcoholic fatty liver disease - NAFLD) in COPD as an underestimated comorbidity (publication 1), although it probably has important prognostic implications (publication 2). COPD is also associated with non-respiratory symptoms such as pain, the variations and locations of which are poorly known during and after exacerbation (publication 3). Treatment of this pain with opiates may have a specifically deleterious effect in this population (publication 5). The challenges of health system reform with cost-effectiveness optimization encourage the development and validation of new methods of outpatient management of exacerbations (publication 6).In a first part of the thesis we explore the epidemiological links between COPD and NAFLD. We will also explore the consequences of such an association on the cardiovascular outcome of patients in the medium term. In a second part, we seek to define the characteristics of pain before and after exacerbation during COPD, the link between pain, anxiety and depression in these patients and the safety of opioids in this fragile population. In the last part, we will discuss the stratification of the risk linked to an exacerbation of COPD, and the possibility of outpatient care for exacerbations of moderate severity.

El objetivo de la presente investigación es determinar la relación entre las variables marketing experiencial y la recompra de cervezas artesanales en los bares por parte de hombres y mujeres entre los 20 a 35 años. De esta manera, poder identificar qué variable tiene mayor correlación con la intención de compra. Esta investigación es de alcance correlacional, ya que busca describir y relacionar los motivos de intención de compra en base a las variables y dimensiones puestas en estudio. Asimismo, se decidió investigar antecedentes, entrevistar a tres expertos en el rubro y realizar dos focus group con el público objetivo. Se halló que las variables y dimensiones de mayor relación con la recompra son la experiencia, el producto, los precios y estilo de vida. Por último, a partir de la investigación, se ha conocido de forma más profunda al público objetivo. Los resultados con mayor relevancia son que el marketing experiencial junto a las acciones de actuaciones, son fundamentales para que el target recompre cervezas artesanales en los bares y que definitivamente, el precio influye en la decisión de compra. / The objective of this research is to determine the relationship between the variables experiential marketing and the repurchase of craft beers in breweries by men and women between the ages of 20 and 35. In this way, to identify which variable has the greatest correlation with the purchase intention. This research is based on a scope of correlational, since it seeks to describe and relate the reasons for purchase intention based on the variables and dimensions put into study. Likewise, it was decided to investigate the background, interviewed three experts in the field and conducted two focus groups with the target audience. It was found that the variables and dimensions most related to the repurchase are the experience, product, prices and the target lifestyle. Finally, based on the research, the target has been known and studied more deeply. The results with greater relevance are that experiential marketing along with the actions of acts, are essential for the target to repurchase craft beers in the bars and that definitely, the price also influences on the purchase decision. / Trabajo de investigación

En los últimos años, el mercado de bebidas alcohólicas “ready to drink” ha crecido a pasos agigantados siendo el segmento dentro de la categoría de bebidas alcohólicas que mayor crecimiento ha tenido. Las empresas y emprendedores han notado esto y quieren ser participes del éxito de las “ready to drink”. Actualmente el mercado lo lideran las marcas Piscano y Smirnoff; sin embargo, las marcas más pequeñas pueden ganar terreno en el punto de venta si usan adecuadamente su etiqueta para comunicar el posicionamiento de la marca y llegar a su público objetivo. Según los expertos y los propios consumidores, el diseño de las etiquetas tiene un margen de mejora en el mercado peruano. Por ello es que se realiza la siguiente investigación, con el fin de conocer los lineamientos a tomar en cuenta para el diseño del packaging y de esta manera hacer más competitivo el mercado “ready to drink” en el Perú. La presente investigación busca relacionar la comunicación del posicionamiento en las etiquetas con la intención de compra. Para ello, se utilizaron técnicas de recolección de datos cuantitativos y cualitativos que luego de ser analizados se ha logrado obtener lineamientos a tomar en cuenta a la hora del diseño de la etiqueta como, los elementos más relevantes de la etiqueta, su influencia con en el consumidor y la intención de compra. / In recent years, the “ready to drink” alcoholic beverage market has grown by leaps and bounds, being the segment within the category of alcoholic beverages that has grown the most. Companies and entrepreneurs have noticed this and want to be part of the success of "ready to drink". The market is currently led by the Piscano and Smirnoff brands; however, smaller brands can gain ground at the point of sale if they properly use their label to communicate brand positioning and reach their target audience. According to experts and consumers, the design of the labels has a margin of improvement in the Peruvian market. That is why the following research is carried out, in order to know the guidelines to be taken into account for the design of the packaging and thus make the “ready to drink” market in Peru more competitive. The present investigation seeks to relate the communication of the positioning in the labels with the intention of purchase. For this, quantitative and qualitative data collection techniques were used that after being analyzed, it has been possible to obtain guidelines to be taken into account when designing the label as, the most relevant elements of the label, its influence with the consumer and the intention of purchase. / Trabajo de investigación

The adverse health and safety consequences of excessive alcohol consumption are a leading problem around the world. Alcoholic beverages are a routine part of socializing in many societies. However, alcohol is also a significant contributor to worldwide morbidity, disability, and mortality. To lessen the harm produced by alcohol, governments have adopted different alcohol control policies. These control policies can be group into four basic strategies: deterrence, prevention, communications and outreach, and alcohol treatment. Among the prevention measures, restricting physical access to alcohol by limiting the alcohol outlets' density, raising the legal age to purchase alcohol, and reducing the affordability of alcohol through taxation have been extensively shown as cost-effective and feasible measures against alcohol-related harms. However, there are still topics related to the affordability of alcohol that have not been investigated. The role of promotional price practices at on-premises alcohol outlets on health and social outcomes, and the effects of policies enacted to prevent these practices on motor vehicle crashes remained an unexplored research topic. The main goals of this dissertation were to summarize evidence regarding the health effects of drink specials and to estimate the effects of policies restricting drink special practices as preventive tools for fatal motor vehicle crashes. Specifically, I summarized the research evidence of the effects of drink special practices on health and social outcomes (Aim 1). I examined the association between drink special laws and alcohol-related fatal motor vehicle crashes contrasting results for two methodological approaches, difference-in-difference-in-differences (Aim 2) and synthetic controls (Aim 3). This dissertation contains five chapters. The introduction in chapter one provides a background review of relevant literature that serves as the conceptual framework for this dissertation and an overview of chapters two, three, four, and five. The systematic review of the literature relevant to Aim 1 is presented in chapter two. This review included studies on the effects that drink specials and drink special laws have on alcohol consumption, binge drinking, and alcohol-related harms. Twelve studies examined the effect of drink specials in seven countries between 1978 and 2018. Consistent evidence supported associations between drink specials and increased alcohol consumption, heavy drinking, alcohol intoxication, and other alcohol-related outcomes. For aims 2 and 3, I examined 36-years of data from the U.S. Department of Transportation, Census Bureau, and NIH’s Alcohol Policy Information System and National Institute on Alcohol Abuse and Alcoholism from 1982 to 2017. In chapter three, I presented results from difference-in-difference-in-differences analyses of the effects of implementing six drink special laws on alcohol- and non-alcohol-related motor vehicle fatal crash rates in the United States (U.S.). I assessed exposure to implementation as any law, number of laws, and each law. Random effects generalized least squares regression models adjusted for the proportion of males in the state, youth involved in fatal crashes, gallons of ethanol per capita among the population age 21 years and older, and autonomy index were fitted across 24 treated and 18 non-treated states. Results revealed that the implementation of any drink special law was associated with reductions in overall and alcohol-related fatal crash rates compared to untreated states. Also, drink special laws mitigated incremental rates of non-alcohol related crashes among treated states with any drink special law compared to untreated states. In chapter four, I presented results from synthetic control analyses for single and multiple treated units. I assessed the association between drink special laws and alcohol-related fatal motor vehicle crashes and adjusted for the same covariates as in chapter three. Results in chapter four indicated that states treated with any drink special law reduced alcohol-related fatal crash rates only in years three, five, and ten post-implementation compared to the synthetic control trend. The effects of any drink special law were more consistent at different times in the post-implementation for reducing non-alcohol-related fatal crash rates than the synthetic control trend. Findings for the number of laws implemented and each drink special laws were mixed. Chapter five presents a synthesis and discussion of findings in chapters two, three, and four, as well as policy recommendations for stakeholders and future research.

Introduction: Non-invasive assessment of steatosis and fibrosis is of growing relevance in non-alcoholic fatty liver disease (NAFLD). 1H-Magnetic resonance spectroscopy (1H-MRS) and the ultrasound-based controlled attenuation parameter (CAP) correlate with biopsy proven steatosis, but have not been correlated with each other so far. We therefore performed a headto- head comparison between both methods. Methods: Fifty patients with biopsy-proven NAFLD and 15 healthy volunteers were evaluated with 1H-MRS and transient elastography (TE) including CAP. Steatosis was defined according to the percentage of affected hepatocytes: S1 5-33%, S2 34–66%, S3 $67%. Results: Steatosis grade in patients with NAFLD was S1 36%, S2 40% and S3 24%. CAP and 1H-MRS significantly correlated with histopathology and showed comparable accuracy for the detection of hepatic steatosis: areas under the receiveroperating characteristics curves were 0.93 vs. 0.88 for steatosis $S1 and 0.94 vs. 0.88 for $S2, respectively. Boot-strapping analysis revealed a CAP cut-off of 300 dB/m for detection of S2-3 steatosis, while retaining the lower cut-off of 215 dB/m for the definition of healthy individuals. Direct comparison between CAP and 1H-MRS revealed only modest correlation (total cohort: r = 0.63 [0.44, 0.76]; NAFLD cases: r = 0.56 [0.32, 0.74]). For detection of F2–4 fibrosis TE had sensitivity and specificity of 100% and 98.1% at a cut-off value of 8.85 kPa. Conclusion: Our data suggest a comparable diagnostic value of CAP and 1H-MRS for hepatic steatosis quantification. Combined with the simultaneous TE fibrosis assessment, CAP represents an efficient method for non-invasive characterization of NAFLD. Limited correlation between CAP and 1H-MRS may be explained by different technical aspects, anthropometry, and presence of advanced liver fibrosis.

Non-alcoholic fatty liver disease (NAFLD) is an increasingly prevalent issue in the modern world, predisposing patients to serious pathology such as cirrhosis and hepatocellular carcinoma. Mitochondrial dysfunction, and in particular, diminished hepatic oxidative phosphorylation (OXPHOS) capacity, have been observed in NAFLD livers, which may participate in NAFLD pathogenesis. To examine the role of OXPHOS in NAFLD, we generated a model of enhanced hepatic OXPHOS using mice with liver-specific transgenic expression of LRPPRC, a protein which activates mitochondrial transcription and augments OXPHOS capacity. When challenged with high-fat feeding, mice with enhanced hepatic OXPHOS were protected from the development of liver steatosis and inflammation, critical components in the pathogenesis of NAFLD. This protection corresponded to increased liver and whole-body insulin sensitivity. Moreover, mice with enhanced hepatic OXPHOS have increased availability of oxidized NAD+, which promotes complete fatty acid oxidation in hepatocytes. Interestingly, mice with enhanced hepatic OXPHOS were also protected from obesogenic effects of long-term high-fat feeding. Consistent with this, enhanced hepatic OXPHOS increased energy expenditure and adipose tissue oxidative gene expression, suggesting a communication between the liver and adipose tissue to promote thermogenesis. Examination of pro-thermogenic molecules revealed altered bile acid composition in livers and serum of LRPPRC transgenic mice. These mice had increased expression of bile acid synthetic enzymes, genes which are induced by NAD+ dependent deacetylase SIRT1 activation of the transcriptional co-regulator PGC-1a. These findings suggest that enhanced hepatic OXPHOS transcriptionally regulates bile acid synthesis and dictates whole-body energy expenditure, culminating in protection from obesity.