Cardiovascular-related accidents such as stroke are currently ranked as the second leading cause of death worldwide, and the risk of stroke increases dramatically with age. Aging results in structural and functional alterations of the oligodendrocytes which lead to loss of neuronal connectivity, cognitive deficits, and increased susceptibility to ischemic damage. Here, we have carried out proteomic profiling of MO3.13 oligodendrocyte cells following treatment with NMDA channel blocker MK-801 to increase our understanding of the mechanisms involved in brain aging, as well as those which render it more susceptible to ischemic damage. The main objective was to identify potential biomarkers which could be used to track disease or therapeutic effects.Psychiatric disorders such as major depression are linked to early mortality, and patients affected by these conditions are at an increased risk of developing other diseases that are characteristic of the old and very old. Antidepressants are prescribed in the treatment of depression, although the mechanism of how they exert their therapeutic effects is only partly understood. To shed further light on their mode of action, this chapter presents a protocol using two-dimensional differential in-gel electrophoresis (2D-DIGE) combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry for identifying a proteomic signature in guinea pig brains after treatment with the antidepressant, fluoxetine. As this signature pointed toward changes in synaptic structure, we also present a protocol for Western blot analysis targeting selected proteins identified by the combined 2D-DIGE-MALDI-TOF mass spectrometry procedure. Such validation experiments are critical for the translation of putative biomarkers into preclinical and clinical studies.This chapter describes a protocol for proteomic profiling of the rat hippocampal proteome using a combination of liquid chromatography tandem mass spectrometry (LC-MS/MS) and data analysis to determine the cellular location of the identified proteins.  https://www.selleckchem.com/products/tpen.html  In the example given, many of these proteins were localized in the plasma membrane and nucleus. These could be of interest in further studies of neurological and neurodegenerative disorders linked with hippocampal dysfunction, such as aging, major depression, and Alzheimer's disease.Aging of the brain can result in excessive glucocorticoid secretion, potentially due to chronic stress and related situations. This can lead to dysfunction of brain areas involved in control of the hypothalamic-pituitary adrenal axis, growth, and metabolism, as well as areas associated with cognition and mood regulation. This chapter presents a protocol for two-dimensional differential in-gel electrophoresis (2D-DIGE) analysis of hypothalamus and hippocampus tissue obtained from mice following exposure to high levels of corticosterone for 14 days. The chapter also presents a method for identification of the affected proteins in these brain regions using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.This chapter describes the application of multiplex immunoassay of hormones which are known to be present at different levels during aging and may therefore be precipitous in age-related diseases. As an example, we have analyzed serum from 12-week-old rats using multiplexes for the hormones insulin, leptin, growth hormone, corticosterone, and testosterone, which have been implicated in sarcopenia. The same technique can be used in the investigation of other sarcopenia biomarkers as well as in other disease cases involving both clinical and preclinical biomarker studies.Apolipoproteins have important structural and functional roles in several lipoprotein particles. Apolipoproteins regulate lipid metabolism, adipose tissue, and energy production and serve major regulatory roles in both pre- and pro-atherosclerotic mechanisms. They are also involved in protective mechanisms against atherosclerotic plaques. Therefore, accurate quantification of apolipoproteins may serve as a crucial biomarker for cardiovascular diseases. However, most apolipoproteins cannot be detected using standard clinical immunoassays, and multiplexing is not available for some species of apolipoproteins. Herein, we describe a highly robust and quantitative method using liquid chromatography coupled to tandem mass spectrometry to quantify apolipoproteins in plasma. This methodology may add clinical value for profiling cardiovascular risk in vulnerable individuals and enable monitoring of apolipoprotein levels in plasma following intervention strategies.Sarcopenia is defined as an age-related reduction in muscle mass and performance. Some of the most important risk factors include advanced age, malnutrition, and sedentary lifestyle. The aim of this study was to investigate the association of food intake and physical activity with body composition, muscle strength, and muscle function in a cross-sectional study of postmenopausal women. This analysis gave a positive association between physical activity and handgrip strength, calorie intake and muscle function, protein intake and fat-free mass percentage, and total fat intake and fat mass percentage. In addition, there were negative associations found between carbohydrate intake and muscle function, as well as total fat intake and fat-free mass percentage. This chapter presents a protocol for the study setup along with measurements of physical activity, handgrip strength, nutrient intake, and fat-free mass percentage.Sarcopenia is the loss of muscle strength and muscle mass with aging and is one of the major risk factors for metabolic diseases. Cross-sectional studies have shown that vitamin D is associated with sarcopenia in both men and women. We investigated the effect of vitamin D supplementation over 12 weeks on muscle strength, muscle function, and body composition in middle-aged women in randomized double-blind placebo-controlled trial format. This revealed a significant difference in serum 25-hydroxyvitamin D levels between the intervention and placebo groups. In addition, handgrip strength was improved, and the timed get up and go (TGUG) test and body fat content were decreased. This chapter presents a protocol for trial setup involving measurement of vitamin D levels, handgrip strength, the TGUT test, and body composition as readouts.