Saccharomyces Cerevisiae as a biotechnological tool for ageing research:studies on translation and metabolism

The yeast Saccharomyces cerevisiae is an important model organism for the study of cell biology. The similarity between yeast and human genes and the conservation of fundamental pathways means it can be used to investigate characteristics of healthy and diseased cells throughout the lifespan. Yeast is an equally important biotechnological tool that has long been the organism of choice for the production of alcoholic beverages, bread and a large variety of industrial products. For example, yeast is used to manufacture biofuels, lubricants, detergents, industrial enzymes, food additives and pharmaceuticals such as anti-parasitics, anti-cancer compounds, hormones (including insulin), vaccines and nutraceuticals. Its function as a cell factory is possible because of the speed with which it can be grown to high cell yields, the knowledge that it is generally recognized as safe (GRAS) and the ease with which metabolism and cellular pathways, such as translation can be manipulated. In this thesis, these two pathways are explored in the context of their biotechnological application to ageing research: (i) understanding translational processes during the high-yielding production of membrane protein drug targets and (ii) the manipulation of yeast metabolism to study the molecule, L-carnosine, which has been proposed to have anti-ageing properties. In the first of these themes, the yeast strains, spt3?, srb5?, gcn5? and yTHCBMS1, were examined since they have been previously demonstrated to dramatically increase the yields of a target membrane protein (the aquaporin, Fps1) compared to wild-type cells. The mechanisms underlying this discovery were therefore investigated. All high yielding strains were shown to have an altered translational state (mostly characterised by an initiation block) and constitutive phosphorylation of the translational initiation factor, eIF2a. The relevance of the initiation block was further supported by the finding that other strains, with known initiation blocks, are also high yielding for Fps1. A correlation in all strains between increased Fps1 yields and increased production of the transcriptional activator protein, Gcn4, suggested that yields are subject to translational control. Analysis of the 5´ untranslated region (UTR) of FPS1 revealed two upstream open reading frames (uORFs). Mutagenesis data suggest that high yielding strains may circumvent these control elements through either a leaky scanning or a re-initiation mechanism. In the second theme, the dipeptide L-carnosine (ß-alanyl-L-histidine) was investigated: it has previously been shown to inhibit the growth of cancer cells but delay senescence in cultured human fibroblasts and extend the lifespan of male fruit flies. To understand these apparently contradictory properties, the effects of L-carnosine on yeast were studied. S. cerevisiae can respire aerobically when grown on a non-fermentable carbon source as a substrate but has a respiro-fermentative metabolism when grown on a fermentable carbon source; these metabolisms mimic normal cell and cancerous cell metabolisms, respectively. When yeast were grown on fermentable carbon sources, in the presence of L-carnosine, a reduction in cell growth and viability was observed, which was not apparent for cells grown on a non-fermentable carbon source. The metabolism-dependent mechanism was confirmed in the respiratory yeast species Pichia pastoris. Further analysis of S. cerevisiae yeast strains with deletions in their nutrient-sensing pathway, which result in an increase in respiratory metabolism, confirmed the metabolism-dependent effects of L-carnosine.

Engineering of a novel Saccharomyces cerevisiae wine strain with a respiratory phenotype at high external glucose concentrations

The recently described respiratory strain Saccharomyces cerevisiae KOY.TM6*P is, to our knowledge, the only reported strain of S. cerevisiae which completely redirects the flux of glucose from ethanol fermentation to respiration, even at high external glucose concentrations (27). In the KOY.TM6*P strain, portions of the genes encoding the predominant hexose transporter proteins, Hxt1 and Hxt7, were fused within the regions encoding transmembrane (TM) domain 6. The resulting chimeric gene, TM6*. encoded a chimera composed of the amino-terminal half of Hxt1 and the carboxy-terminal half of Hxt7. It was subsequently integrated into the genome of an hxt null strain. In this study, we have demonstrated the transferability of this respiratory phenotype to the V5 hxt1-7Δ strain, a derivative of a strain used in enology. We also show by using this mutant that it is not necessary to transform a complete hxt null strain with the TM6* construct to obtain a nonethanol-producing phenotype. The resulting V5.TM6*P strain, obtained by transformation of the V5 hxt1-7Δ strain with the TM6* chimeric gene, produced only minor amounts of ethanol when cultured on external glucose concentrations as high as 5%. Despite the fact that glucose flux was reduced to 30% in the V5.TM6*P strain compared with that of its parental strain, the V5.TM6*P strain produced biomass at a specific rate as high as 85% that of the V5 wild-type strain. Even more relevant for the potential use of such a strain for the production of heterologous proteins and also of low-alcohol beverages is the observation that the biomass yield increased 50% with the mutant compared to its parental strain. Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Eastern Orthodox Christian fasting in northeastern Pennsylvania

The purpose of this research was to examine the fasting practices of Eastern Orthodox Christians (EOCs) in northeastern Pennsylvania. Fasting, according to Eastern Orthodox Church doctrine, is primarily abstinence from meat, dairy products, fish and certain other foods during Easter Lent and other periods, for approximately 180 days annually. Goals were to discern what EOCs consider their fasting rules to be, their actual fasting practices, what factors influence this practice and the relationship of fasting to nutrition. Methodology included 29 months of ethnographic fieldwork at local parishes, content analysis of local written materials and semi-structured interviews of 58 core church members. A pile sort was conducted whereby subjects classified various foods according to fasting or non-fasting status and then sorted the fasting foods into a hierarchy of avoidance. Data were analyzed using ANTHROPAC and NVivo software. Results included identification of a cognitive hierarchy of avoidance, with meat the most important to avoid, followed by dairy and alcoholic beverages. An important finding was the differences in subjects' knowledge of Church doctrine and a wide variation in their actual fasting practices. Contrary to Church doctrine, fish was not usually perceived as a food to abstain from. A historic Byzantine Catholic presence in the area (with a different fasting doctrine), family members who did not fast, and health concerns were some factors that affected fasting practices. A conclusion is that while meat, dairy and alcoholic beverages were usually categorized as foods to avoid during fasts, it is not possible to generalize with regard to actual practices or the impact of fasting on nutrition, due to individual variation. It was demonstrated that qualitative data could provide information that can be crucial to know prior to conducting quantitative nutrition research or counseling. Findings of this study suggest that one cannot assume subjects who belong to a given religion that has prescribed food avoidance practices are following them homogeneously and/or according to official doctrine. ^

Managers' Perception of Alcohol Service Training Programs

In order to serve alcoholic beverages in a responsible manner, most managers turn to alcohol server training programs to provide effective standardized training for the alcohol server. Whether or not the programs are perceived by hospitality managers as effective has not been established. The author presents the results of a study of Oklahoma hospitality managers who perceive alcohol server training as becoming less effective.

Current Status of Collectibility of Gaming-Related Credit Dollars

In her piece entitled - Current Status Of Collectability Of Gaming-Related Credit Dollars - Ruth Lisa Wenof, Graduate Student at Florida International University initially states: “Credit is an important part of incentives used to lure gamblers to gaming establishments. However, a collection problem exists in casinos retrieving gaming-related credit losses of individuals living in states where gambling is illegal. The author discusses the history of this question, citing recent cases related to Atlantic City.” This author’s article is substantially laden with legal cases associated with casinos in New Jersey; Atlantic City to be exact. The piece is specific to the segment of the gaming industry that the title suggests, and as such is written in a decidedly technical style. “Legalized casino gaming, which was approved by the citizens of New Jersey on November 8, 1976, has been used as a unique tool of urban redevelopment for Atlantic City,” Wenof says in providing some background on this ‘Jersey shore municipality. “Since Resorts International opened its casino…revenues from gambling have increased rapidly. Resorts' gross win in 1978 was $134 million,” Wenof says. “Since then, the combined gross win of the city's 11 casinos has been just shy of $7.5 billion.” The author points out that the competition for casino business is fierce and that credit dollars play an integral role in soliciting such business. “Credit plays a most important part in every casino hotel. This type of gambler is given every incentive to come to a particular hotel,” says the author. “Airplanes, limousines, suites, free meals, and beverages all become a package for the person who can sign a marker. The credit department of a casino is similar to that of a bank. A banker who loans money knows that it must be paid back or his bank will fail. This is indeed true of a casino,” Wenof warns in outlining the potential problem that this article is fundamentally designed around. In providing further background on credit essentials and possible pitfalls, Wenof affords: “…on the Casino Control Act the State Commission of Investigation recommended to the legislature that casinos should not be allowed to extend credit at all, by reason of a concern for illicit diversion of revenues, which is popularly called skimming within the industry…” Although skimming is an after-the-fact problem, and is parenthetic to loan returns, it is an important element of the collective [sic] credit scheme. “A collection problem of prime importance is if a casino can get back gaming-related credit dollars advanced by the casino to a gambler who lives in a state where gambling is illegal,” is a central factor to consider, Wenof reveals. This is a primary focus of this article. Wenof touches on the social/societal implications of gambling, and then continues the discussion by citing a host of legal cases pertaining to debt collection.

Manager's Perceptions of Alcohol Server Staffing and Training Methods

Over-consumption of alcoholic beverages is a concern of managers of hotels and motels with a club/lounge, restaurant, and tavern. The authors surveyed members of two industry associations in Oklahoma to ascertain alcohol server training methods and managers' perception of the value of such programs.

Beverage Ethics: Education for Alcohol Responsibility

The study looked at the processes in the development of an alcohol responsibility program for post-secondary students in the service management major at the University of Tennessee: he program has been certified by the State of Tennessee to satisfy the Alcohol Beverage Commission requirement for server training related to the handling and service of alcoholic beverages. A managerial viewpoint was adopted so as to provide the greatest benefit to service management graduate.

Adult Alternatives for Social Drinking: A Direction

In their survey/study - Adult Alternatives for Social Drinking: A Direction - by John Dienhart and Sandra Strick, Assistant Professors, Department of Restaurant, Hotel and Institutional Management, Purdue University, Dienhart and Strick begin with: “Changes in consumer habits have brought about a change in the business of selling alcoholic drinks and have impacted upon hotel food and beverage operations. The authors surveyed a sample of hotel corporate food and beverage directors to ascertain how they are handling this challenge.” Dienhart and Strick declare that the alcoholic beverage market, sale and consumption thereof, has taken a bit of a hit in contemporary society. “Even to the casual observer, it's obvious that the bar and beverage industry has undergone a great deal of change in the past few years,” say the authors. “Observations include a change in the types of drinks people are ordering, as well as a decrease in the number of drinks being sold,” they qualify. Dienhart and Strick allude to an increase in the federal excise tax, attacks from alcohol awareness groups, the diminished capacity of bars and restaurants to offer happy hours, increased liability insurance premiums as well as third-party liability issues, and people’s awareness of their own mortality as some of the reasons for the change. To quantify some empirical data on beverage consumption the Restaurant, Hotel, and Institutional Management Department of Purdue University conducted a study “… to determine if observed trends could be documented with hard data.” In regards to the subject, the study asks and answers a lot of interesting questions with the results presented to concerned followers via percentages. Typical of the results are: “When asked whether the corporation experienced a change in alcoholic sales in the past year, 67 percent reported a decrease in the amount of alcohol sold.” “Sixty-two percent of the respondents reported an increase in non-alcoholic sales over the past year. The average size of the increase was 8 percent. What Dienhart and Strick observe is that the decrease in alcoholic beverage consumption has resulted in a net increase for non-alcoholic beverage consumption. What are termed specialty drinks are gaining a foothold in the market, say the authors. “These include traditional cocktails made with alcohol-free products, as well as creative new juice based drinks, cream based drinks, carbonated beverages, and heated drinks,” say Dienhart and Strick by way of citation . Another result of the non-alcoholic consumption trend is the emergence of some novel marketing approaches by beer, wine, and spirits producers, including price increases on their alcohol based beverages as well as the introduction of faux alcoholic drinks like non-alcoholic beer and wine. Who or what is the big winner in all of this? That distinction might go to bottled water!

Comparison of coronary heart disease risk factors in foreign -born and United States-born Afro Caribbean Americans and African Americans in south Florida

In a cross-sectional study design, risk factors for coronary heart disease (CHD) were evaluated in three groups: 66 Afro Caribbeans (FBCA) living in the US for less than 10 years, 62 US-born Afro Caribbean (USBCA) and 61 African American (AA) adults (18–40 years), with equal numbers of males and females in each group. Socio-demographic, dietary, anthropometric and blood pressure data were collected. Fasting blood glucose, blood lipids and high-sensitivity C-reactive protein (hs-CRP) were determined. ^ The USBCA and AA participants compared to the FBCA participants consumed significantly (p < 0.05) more mean total fat (g) (66.3 ± 41.7 and 73.0 ± 47.8 vs. 52.8 ± 32.3), saturated fat (g) (23.1 ± 14.9 and 24.9 ± 15.8 vs. 18.6 ± 11.5), percent energy from fat (%) (33.1 ± 6.5 and 31.4 ± 6.4 vs. 29.3 ± 6.8), fat servings (1.8 ± 1.2 and 1.5 ± 1.0 vs. 1.2 ± 0.9), dietary cholesterol (mg) (220.4 ± 161.9 and 244.1 ± 155.0 vs. 168.8 ± 114.0) and sodium (mg) (2245.2 ± 1238.3 and 2402.6 ± 1359.3 vs. 1838.0 ± 983.4) and less than 2 servings of fruits per day (%) (86.9 and 94.9 vs. 78.5). These differences were more pronounced in males compared to females and remained after correcting for age. Also, the percentages of USBCA and AA participants who were obese (17.1% and 23.0%, respectively) were significantly (p < 0.05) higher compared to FBCA (7.6%) participants. More USBCA and AA than FBCA individuals smoked cigarettes (4.8% and 6.6% vs. 0.0%) and consumed alcoholic beverages (29.0% and 50.8% vs. 24.2%). The mean hs-CRP level of the AA participants (2.2 ± 2.7 mg/L) was significantly (p < 0.01) higher compared to the FBCA (1.1 ± 1.3 mg/L) and USBCA (1.3 ± 1.6 mg/L) participants. ^ The FBCA participants had a better CHD risk profile than the USBCA and AA participants. Focus should be placed on the ethnic and cultural differences in a population to better understand the variations in health indicators among different ethnic groups of the same race. This focus can provide healthcare professionals and policy planners with the opportunity to develop culturally sensitive programs and strategies for the improvement of health outcomes. ^

Psychological and cognitive factors that influence post -exercise energy intake in normal weight and overweight sedentary males

Physical activity is recommended to facilitate weight management. However, some individuals may be unable to successfully manage their weight due to certain psychological and cognitive factors that trigger them to compensate for calories expended in exercise. The primary purpose of this study was to evaluate the effect of moderate-intensity exercise on lunch and 12-hour post-exercise energy intake (PE-EI) in normal weight and overweight sedentary males. Perceived hunger, mood, carbohydrate intake from beverages, and accuracy in estimating energy intake (EI) and energy expenditure (EE) were also assessed. The study consisted of two conditions, exercise (treadmill walking) and rest (sitting), with each participant completing each condition, in a counterbalanced-crossover design on two days. Eighty males, mean age 30 years (SD=8) were categorized into five groups according to weight (normal-/overweight), dietary restraint level (high/low), and dieting status (yes/no). Results of repeated measures, 5x2 ANOVA indicated that the main effects of condition and group, and the interaction were not significant for lunch or 12-hour PE-EI. Among overweight participants, dieters consumed significantly (p<0.05) fewer calories than non-dieters at lunch (M=822 vs. M=1149) and over 12 hours (M=1858 vs. M =2497). Overall, participants’ estimated exercise EE was significantly (p<0.01) higher than actual exercise EE, and estimated resting EE was significantly (p<0.001) lower than actual resting EE. Participants significantly (p<0.001) underestimated EI at lunch on both experimental days. Perceived hunger was significantly (p<0.05) lower after exercise (M=49 mm, SEM=3) than after rest (M=57 mm, SEM=3). Mood scores and carbohydrate intake from beverages were not influenced by weight, dietary restraint, and dieting status. In conclusion, a single bout of moderate-intensity exercise did not influence PE-EI in sedentary males in reference to weight, dietary restraint, and dieting status, suggesting that this population may not be at risk for overeating in response to exercise. Therefore, exercise can be prescribed and used as an effective tool for weight management. Results also indicated that there was an inability to accurately estimate EI (ad libitum lunch meal) and EE (60 minutes of moderate-intensity exercise). Inaccuracies in the estimation of calories for EI and EE could have the potential to unfavorably impact weight management.

Avaliação da efetividade do Programa de Alimentação do Trabalhador

This study analyzed the Worker’s Healthy Eating Program in Rio Grande do Norte state (RN) to assess its possible impact on the nutritional status of the workers benefitted. To that end, we conducted a cross-sectional observational prospective study based on a multistage stratified random sample comparing 26 small and medium-sized companies from the Manufacturing Sector (textiles, food and beverages, and nonmetallic minerals) of RN, divided into two equal groups (WFP and Non WFP). Interviews were conducted at each company by trained interviewers from Tuesday to Saturday between September and December 2014. Data were collected on the company (characterization and information regarding the program’s desired results) and workers (personal and professional information, anthropometrics, health, lifestyle and food consumed the previous day). Population estimates were calculated for RN on the characteristics of workers and the study variables. The main variable was BMI. The secondary variables were waist circumference (WC), nutritional diagnosis, calorie intake, blood pressure, metabolic variables and lifestyle indicators. The statistical method used was hierarchical mixed effects linear regression for interval variables and hierarchical mixed effects logistic regression for binary variables. The variables measured in ordinal scales were analyzed by ordinal logistic regression adjusted for correlated variables, adopting robust standard errors. The results for interval variables are presented as point estimates and their 95% confidence intervals; and as odds-ratios and their 95% confidence intervals for binary variables. The Fisher’s exact and Student’s t-tests were used for simple comparisons between proportions and means, respectively. Differences were considered statistically significant at p<0.05. A total of 1069 workers were interviewed, of which 541 were from the WFP group and 528 from the Non WFP group. Subjects were predominantly males and average age was 34.5 years. Significant intergroup differences were observed for schooling level, income above 1 MW (minimum wage) and specific training for their position at the company. The results indicated a significant difference between the BMI of workers benefitted, which was on average 0.989 kg/m2 higher than the BMI of workers from the Non WFP group (p=0.002); and between the WC, with the waist circumference of WFP group workers an average of 1.528 cm larger (p<0.05). Higher prevalence of overweight and obesity (p<0.001) and cardiovascular risk (p=0.038) were recorded in the WFP group. Tests on the possible effect of the WFP on health (blood pressure and metabolic indicators) and lifestyle indicators (smoking, alcohol consumption and exercise) were not significant. With respect to worker’s diets, differences were significant for consumption of saturated fat (lunch and daily intake), salt (lunch, other meals and daily intake) and proteins (other meals and daily intake), with higher consumption of these nutrients in the WFP group. The study showed a possible positive impact of the WFP on nutritional status (BMI and WC) among the workers benefitted. No possible effects of the program were observed for the lifestyle indicators studied. Workers benefitted consumed less salt, saturated fat and protein. The relevance of the WFP is recognized for this portion of society and it is understood that, if the program can reach and impact those involved, the development of educational initiatives aimed at nutritional and food safety may also exert a positive influence.

Herb and tea seller at Women's Event in Las Cruces, New Mexico

General note: Title and date provided by Bettye Lane.

Catabolic flexibility of mammalian-associated lactobacilli

Metabolic flexibility may be generally defined as “the capacity for the organism to adapt fuel oxidation to fuel availability”. The metabolic diversification strategies used by individual bacteria vary greatly from the use of novel or acquired enzymes to the use of plasmid-localised genes and transporters. In this review, we describe the ability of lactobacilli to utilise a variety of carbon sources from their current or new environments in order to grow and survive. The genus Lactobacillus now includes more than 150 species, many with adaptive capabilities, broad metabolic capacity and species/strain variance. They are therefore, an informative example of a cell factory capable of adapting to new niches with differing nutritional landscapes. Indeed, lactobacilli naturally colonise and grow in a wide variety of environmental niches which include the roots and foliage of plants, silage, various fermented foods and beverages, the human vagina and the mammalian gastrointestinal tract (GIT; including the mouth, stomach, small intestine and large intestine). Here we primarily describe the metabolic flexibility of some lactobacilli isolated from the mammalian gastrointestinal tract, and we also describe some of the food-associated species with a proven ability to adapt to the GIT. As examples this review concentrates on the following species - Lb. plantarum, Lb. acidophilus, Lb. ruminis, Lb. salivarius, Lb. reuteri and Lb. sakei, to highlight the diversity and inter-relationships between the catabolic nature of species within the genus.

Relationships Between Current and Past Binge Drinking and Systolic Blood Pressure in Young Adults.

PURPOSE: Heavy episodic (i.e., "binge") drinking (i.e., ≥five drinks/occasion) is highly prevalent among young adults; those who binge do so four times per month on average, consuming nine drinks on average on each occasion. Although it is well established that chronic heavy drinking (≥two alcoholic beverages per day) increases the risk of hypertension, the relationship between binge drinking and blood pressure is not well described. Our aim was to describe the relationship between frequency of binge drinking, both current (at age 24 years) and past (at age 20 years), and systolic blood pressure (SBP) at age 24 years. METHODS: Participants (n = 756) from the longitudinal Nicotine Dependence in Teens study reported alcohol consumption at ages 20 and 24 years and had SBP measured at age 24 years. We examined the association between binge drinking and SBP using multiple linear regression, controlling for sex, race/ethnicity, education, monthly drinking in high school, cigarette smoking, and body mass index. RESULTS: Compared to nonbinge drinkers, SBP at age 24 years was 2.61 [.41, 4.82] mm Hg higher among current monthly bingers and 4.03 [1.35, 6.70] mm Hg higher among current weekly bingers. SBP at age 24 years was 2.90 [.54, 5.25] mm Hg higher among monthly bingers at age 20 years and 3.64 [.93, 6.35] mm Hg higher among weekly bingers at age 20 years, compared to nonbinge drinkers. CONCLUSIONS: Frequent binge drinking at ages 20 and 24 years is associated with higher SBP at age 24 years and may be implicated in the development of hypertension.