The model may also be suitable for the future determination of muscarinic receptor binding affinities, which will be valuable as part of future estimations of anticholinergic impact in the human brain. Another vital component of human research into anticholinergic impact in the future would be the in vitro modelling of the role of the blood brain barrier in the systemic anticholinergic drug penetration into the CNS, as well as that of any metabolites formed by the liver or other biotransforming organs. Clinically, anticholinergic agents differ in their abilities to enter the brain and there is evidence that the brain microvessel robustness and permeability is impaired by age and infirmity The NT2.N/A co-cultures could be combined with advanced stem cell-derived human models of the BBB in bioreactor systems to further enhance clinical predictability in terms of anticholinergic drug impact. Future work will aim to examine a greater number of drugs on the ACB scale, alone and in combination of two or more, as this is well within the capabilities of the flexible perfusion system employed. Consultation with experts in the various fields of the primary care of the elderly, such as geriatric psychiatrists, will reveal the most relevant drug combinations worthy of study that 2-MPMDQ exhibit the most anticholinergic effects at clinically relevant doses and CNS free drug concentrations, with the overall aim of establishing a reliably predictive model. Whilst there are a large potential number of possible anticholinergic drug combinations used in clinical practice to investigate, such efforts would be rewarded in terms of the discovery of which new and older drug combinations cause the most damage to the CNS cholinergic health of the elderly. The Sir2 or sirtuin family of class III deaceatylases differs from class I and II histone deacetylases by their sequences and structure. Sirtuins are evolutionarily conserved NAD + -dependent protein deacetylases and adenosine diphosphate -ribosylases. Seven NAD + -dependent HDAC proteins were recognized in mammalians, SIRT1-7 differs in the subcellular localization, substrate specificities, and functions. Sirtuin catalyze the deacetylation of lysine residues on histones and various proteins, resulting in a 5-BDBD deacetylated product as nicotinamide, and O-acetyl-ADP-ribose. The catalytic core of sirtuins, conserved from bacteria to human with variable N- and C-terminals, contains approximately 250 amino acids. The catalytic domain consists of a large typical Rossmann fold or the classic pyridine dinucleotide binding fold, and a small domain composed of residues from two insertions within the Rossman fold, one comprising a zinc-binding module that contains a structural zinc atom coordinated by 4 invariant cysteine��s, and the other forming a helical module that includes a flexible loop. The protein and NAD + co-substrates bind in a cleft between the large and small domains. The members of Sirtuin family play an important role in biological processes, such as life span regulation, fat metabolization in human cells, insulin secretion, cellular response to stress, axonal degeneration, basal transcription factor activity, regulationg enzyme activity, rDNA recombination, and switching between morphological states in Candida, and apoptosis, there has been substantial progress in uncovering the chemical and structural details of these fascinating enzymes.