Towards a Large-Scale Assessment of the Relationship between Biological and Chronological Aging: The INSPIRE Mouse Cohort.

Aging is the major risk factor for the development of chronic diseases. After decades of research focused on extending lifespan, current efforts seek primarily to promote healthy aging. Recent advances suggest that biological processes linked to aging are more reliable than chronological age to account for an individual's functional status, i.e. frail or robust. It is becoming increasingly apparent that biological aging may be detectable as a progressive loss of resilience much earlier than the appearance of clinical signs of frailty. In this context, the INSPIRE program was built to identify the mechanisms of accelerated aging and the early biological signs predicting frailty and pathological aging. To address this issue, we designed a cohort of outbred Swiss mice (1576 male and female mice) in which we will continuously monitor spontaneous and voluntary physical activity from 6 to 24 months of age under either normal or high fat/high sucrose diet. At different age points (6, 12, 18, 24 months), multiorgan functional phenotyping will be carried out to identify early signs of organ dysfunction and generate a large biological fluids/feces/organs biobank (100,000 samples). A comprehensive correlation between functional and biological phenotypes will be assessed to determine: 1) the early signs of biological aging and their relationship with chronological age; 2) the role of dietary and exercise interventions on accelerating or decelerating the rate of biological aging; and 3) novel targets for the promotion of healthy aging. All the functional and omics data, as well as the biobank generated in the framework of the INSPIRE cohort will be available to the aging scientific community. The present article describes the scientific background and the strategies employed for the design of the INSPIRE Mouse cohort.

Identification of the ectoenzyme CD38 as a marker of committed preadipocytes.

BACKGROUND/OBJECTIVES: Characterisation of the adipocyte cellular lineage is required for a better understanding of white adipose tissue homoeostasis and expansion. Although several studies have focused on the phenotype of the most immature adipocyte progenitors, very few tools exist to identify committed cells. In haematopoiesis, the CD38 ectoenzyme is largely used to delineate various stages of stem cell lineage commitment. We hypothesise that this marker could be used to identify committed preadipocytes. METHODS: Complementary strategies including flow cytometry, cell-sorting approaches, immunohistochemistry and primary cultures of murine adipose progenitors isolated from different fat pads of control or high-fat diet exposed C57BL/6 J mice were used to determine the molecular expression profile, proliferative and differentiation potentials of adipose progenitors expressing the CD38 molecule. RESULTS: We demonstrate here that a subpopulation of CD45- CD31- CD34+ adipose progenitors express the cell surface protein CD38. Using a cell-sorting approach, we found that native CD45- CD31- CD34+ CD38+ (CD38+) adipose cells expressed lower CD34 mRNA and protein levels and higher levels of adipogenic genes such as Pparg, aP2, Lpl and Cd36 than did the CD45- CD31- CD34+ CD38- (CD38-) population. When cultivated, CD38+ cells displayed reduced proliferative potential, assessed by BrdU incorporation and colony-forming unit assays, and greater adipogenic potential. In vitro, both CD38 mRNA and protein levels were increased during adipogenesis and CD38- cells converted into CD38+ cells when committed to the adipogenic differentiation programme. We also found that obesity development was associated with an increase in the number of CD38+ adipose progenitors, this effect being more pronounced in intra-abdominal than in subcutaneous fat, suggesting a higher rate of adipocyte commitment in visceral depots. CONCLUSIONS: Together, these data demonstrate that CD38 represents a new marker that identifies committed preadipocytes as CD45- CD31- CD34low CD38+ cells.

Comparison of two online flocculation monitoring techniques for predicting turbidity removal by granular media filtration.

Particulate matter removal in drinking water treatment via direct granular filtration requires specific flocculation conditions (a process typically termed 'high energy flocculation'). Predicting filtered water turbidity based on flocculated water characteristics remains difficult. This study has sought to establish a relationship between filtered water turbidity and the flocculated water characteristics. Flocculation oflow-turbidity raw water was evaluated online using a Photometric Dispersion Analyser (PDA) and a Dynamic Particle Analyser in a modified jar test followed by a bench-scale anthracite filter. Coagulants used were alum, PASS100 and ferric sulphate, in addition to a polydiallyldimethylammonium chloride (polyDADMAC) cationic polymer. They were dosed in warm and cold waters, and flocculated with intensities (G) from 0 to 100 s(-1). Of the two instruments selected to analyse flocculation performance, the Dynamic Particle Analyser was shown to be the most sensitive, detecting small changes in floc growth kinetics and even floc growth under low flocculation conditions which remained undetected by the PDA. Floc size was shown to be insufficient in predicting particulate matter removal by direct granular filtration as measured by turbidity, although a threshold d(v) value (50 microm) could be identified for the test conditions evaluated in this project, above which turbidity was systematically lower than 0.2 NTU.

Plasticity of adipose tissue: a promising therapeutic avenue in the treatment of cardiovascular and blood diseases?

The adipose tissue represents a large amount of adult tissues. For long time, it was considered as a poorly active overgrown and undesirable tissue even if its usefulness was demonstrated in reconstructive surgery. It was studied for its main involvement in energy metabolism and disorders as diabetes and obesity. More recently, its endocrine functions emerged and appeared to play a key role in many physiological situations such as inflammation and immunity. The presence of preadipocytes throughout life was demonstrated using primary culture technology from cells derived from adipose tissue. These cells can display a macrophagic or endothelial potential according to their environment and could be now considered as vascular progenitors. Differentiation of various adipose derived cell subsets towards functional cardiomyocytes, osteoblasts, haematopoietic and neural cells was also obtained in vitro. Altogether, these data emphasise the need to consider with a new look preadipocyte status and adipose tissue biology. These spectacular data, together with the fact that adipose tissue is easy to obtain lead to numerous and promising perspectives in regenerative medicine. They highlight the concept that progenitor cells from adipose tissue constitute an alternative for cells-based strategies designed for the treatment of cardiovascular diseases.

Biological validation of coenzyme Q redox state by HPLC-EC measurement: relationship between coenzyme Q redox state and coenzyme Q content in rat tissues.

The properties of coenzymes Q (CoQ9 and CoQ10) are closely linked to their redox state (CoQox/total CoQ) x 100. In this work, CoQ redox state was biologically validated by high performance liquid chromatography-electrochemical measurement after modulation of mitochondrial electron flow of cultured cells by molecules increasing (rotenone, carbonyl cyanide chlorophenylhydrazone) or decreasing (antimycin) CoQ oxidation. The tissue specificity of CoQ redox state and content were investigated in control and hypoxic rats. In control rats, there was a strong negative linear regression between tissular CoQ redox state and CoQ content. Hypoxia increased CoQ9 redox state and decreased CoQ9 content in a negative linear relationship in the different tissues, except the heart and lung. This result demonstrates that, under conditions of mitochondrial impairment, CoQ redox control is tissue-specific.

Quantitation of tryptophan and tyrosine residues in proteins by fourth-derivative spectroscopy.

A method for quantitation of tryptophan and tyrosine residues in proteins by fourth-derivative ultraviolet spectroscopy is described. The direct quantitation of tryptophan is based on measurement of a tryptophan-specific trough at 292 nm in the fourth derivative of a protein's ultraviolet absorption spectrum. A peak overlapping the tryptophan and tyrosine signatures at A282 is used to quantify tyrosine content. The procedure is accomplished by adding back known quantities of tyrosine to the sample and subtracting the contribution of tryptophan to the A282 peak to obtain an internal calibration curve. This curve is linear, with the ordinate axis intercept relating the quantity (residues/mole) of tyrosine present in the protein. This nondestructive and facile method was used to successfully predict the tryptophan and tyrosine content of a variety of well-characterized proteins. The utility of this method was further demonstrated by resolving the number of tryptophan and tyrosine residues in proteins oxidized by N-bromosuccinimide.

Planning a statewide genetic screening program: defining program functions to achieve desired outcomes.

When responsibility for administering the Genetic Screening Program in Georgia was transferred from an academic institution to state authority in 1982, the need was identified to reassess program planning. Accordingly, a cooperative effort was initiated between the Director of the Genetic Screening Program and representatives of the Centers for Disease Control to define desired program outcomes and the functions that should be performed to achieve these outcomes. This cooperative effort resulted in the development of specific and measurable outcomes for Georgia's Genetic Screening Program. These desired outcomes indicate the degree of reduction in morbidity and mortality associated with genetic diseases the Program is expected to achieve within a specified period of time. The major actions that should be taken to achieve these outcomes were also identified and delineated in sequence using flowchart format. These explicit descriptions of desired program outcomes and the functions necessary to achieve these outcomes provide the Genetic Screening Program Director with a valuable resource to use in planning program activities and assessing the extent to which the Program is successful in achieving its overall goal of reducing morbidity and mortality associated with genetic diseases.

Give your employees a pat on the back.

How does your medical group recognize the long-term service of its employees? Realizing that many of its employees had been under continuous employment for as many as 30 years, the Falls Clinic Professional Association developed a recognition program to honor that faithful service. Employees and physicians were involved in development of the program from many phases--a logo contest was held as an approach to developing a symbol that could be associated with the group and used for the awards. Presentation of the awards has become a tradition that this group eagerly anticipates and is proud to support.