Showing papers in "Sub-cellular biochemistry in 2007"

Anti-inflammatory drugs in the 21st century.

Abstract: Historically, anti-inflammatory drugs had their originsin the serendipitous discovery of certain plants and their extracts being applied for the relief of pain, fever and inflammation. When salicylates were discovered in the mid-19th century to be the active components of Willow Spp., thisenabled these compounds to be synthesized and from this,acetyl-salicylic acid or Aspirin trademark was developed.Likewise, the chemical advances of the 19th–20th centuries leadto development of the non-steroidal anti-inflammatory drugs(NSAIDs), most of which were initially organic acids, but laternon-acidic compounds were discovered. There were two periods ofNSAID drug discovery post-World War 2, the period up to the 1970’swhich was the pre-prostaglandin period and thereafter up to the latter part of the last century in which their effects on prostaglandin production formed part of the screening in thedrug-discovery process. Those drugs developed up to the 1980-late90’s were largely discovered empirically following screening for anti-inflammatory, analgesic and antipyretic activities inlaboratory animal models. Some were successfully developed thatshowed low incidence of gastro-intestinal (GI) side effects (theprincipal adverse reaction seen with NSAIDs) than seen with their predecessors (e.g. aspirin, indomethacin, phenylbutazone); the GIreactions being detected and screened out in animal assays. In the1990’s an important discovery was made from elegant molecular andcellular biological studies that there are two cyclo-oxygenase(COX) enzyme systems controlling the production of prostanoids [prostaglandins (PGs) and thromboxane (TxA2) ]; COX-1that produces PGs and TxA2 that regulategastrointestinal, renal, vascular and other physiological functions, and COX-2 that regulates production of PGs involved in inflammation, pain and fever. The stage was set in the 1990’s forthe discovery and development of drugs to selectively controlCOX-2 and spare the COX-1 that is central to physiological processes whose inhibition was considered a major factor indevelopment of adverse reactions, including those in the GI tract.At the turn of this century, there was enormous commercialdevelopment following the introduction of two new highly selectiveCOX-2 inhibitors, known as coxibs (celecoxib and rofecoxib) whichwere claimed to have low GI side effects. While found to have fulfilled these aims in part, an alarming turn of events tookplace in the late 2004 period when rofecoxib was withdrawn worldwide because of serious cardiovascular events and other coxibs were subsequently suspected to have this adverse reaction, although to a varying degree. Major efforts are currently underwayto discover why cardiovascular reactions took place with coxibs, identify safer coxibs, as well as elucidate the roles of COX-2 andCOX-1 in cardiovascular diseases and stroke in the hope that there may be some basis for developing newer agents (e.g. nitricoxide-donating NSAIDs) to control these conditions

Calcium and Cell Death: The Mitochondrial Connection

Abstract: Physiological stimuli causing an increase of cytosolic free Ca2+ [Ca2+], or the release of Ca2+ from the endoplasmic reticulum invariably induce mitochondrial Ca2+ uptake, with a rise of mitochondrial matrix free [Ca2+] ([Ca2+]m). The [Ca2+]m rise occurs despite the low affinity of the mitochondrial Ca2+ uptake systems measured in vitro and the often limited amplitude of the cytoplasmic [Ca2+]c increases. The [Ca2+]m increase is typically in the 0.2-3 microM range, which allows the activation of Ca2(+)-regulated enzymes of the Krebs cycle; and it rapidly returns to the resting level if the [Ca2+], rise recedes due to activation of mitochondrial efflux mechanisms and matrix Ca2+ buffering. Mitochondria thus accumulate Ca2+ and efficiently control the spatial and temporal shape of cellular Ca2+ signals, yet this situation exposes them to the hazards of Ca2+ overload. Indeed, mitochondrial Ca2+, which is so important for metabolic regulation, can become a death factor by inducing opening of the permeability transition pore (PTP), a high conductance inner membrane channel. Persistent PTP opening is followed by depolarization with Ca2+ release, cessation of oxidative phosphorylation, matrix swelling with inner'membrane remodeling and eventually outer membrane rupture with release of cytochrome c and other apoptogenic proteins. Understanding the mechanisms through which the Ca2+ signal can be shifted from a physiological signal into a pathological effector is an unresolved problem of modern pathophysiology that holds great promise for disease treatment.

Cyclooxygenase-2 (cox-2) and the inflammogenesis of cancer

Abstract: Cohesive scientific evidence from molecular, animal, and human investigations supports the hypothesis that aberrant induction of COX-2 and up-regulation of the prostaglandin cascade play a significant role in carcinogenesis, and reciprocally, blockade of the process has strong potential for cancer prevention and therapy. Supporting evidence includes the following: [1] expression of constitutive COX-2-catalyzed prostaglandin biosynthesis is induced by most cancer-causing agents including tobacco smoke and its components (polycylic aromatic amines, heterocyclic amines, nitrosamines), essential polyunsaturated fatty acids (unconjugated linoleic acid), mitogens, growth factors, proinflammatory cytokines, microbial agents, tumor promoters, and other epigenetic factors, [2] COX-2 expression is a characteristic feature of all premalignant neoplasms, [3] COX-2 expression is a characteristic feature of all malignant neoplasms, and expression intensifies with stage at detection and cancer progression and metastasis, [4] all essential features of carcinogenesis (mutagenesis, mitogenesis, angiogenesis, reduced apoptosis, metastasis, and immunosuppression) are linked to COX-2-driven prostaglandin (PGE-2) biosynthesis, [5] animal studies show that COX-2 up-regulation (in the absence of genetic mutations) is sufficient to stimulate the transformation of normal cells to invasive cancer and metastatic disease, [6] non-selective COX-2 inhibitors, such as aspirin and ibuprofen, reduce the risk of human cancer and precancerous lesions, and [7] selective COX-2 inhibitors, such as celecoxib, reduce the risk of human cancer and precancerous lesions at all anatomic sites thus far investigated. Results confirming that COX-2 blockade is effective for both cancer prevention and therapy have been tempered by observations that some COX2 inhibitors pose a risk to the cardiovascular system, and more studies are needed in order to determine if certain of these drugs can be taken at dosages that prevent cancer without increasing cardiovascular risk. It is emphasized that the "inflammogenesis model of cancer" is not mutually exclusive and may in fact be synergistic with the accumulation of somatic mutations in tumor suppressor genes and oncogenes or epigenetic factors in the development of cancer.

Cerebral creatine deficiency syndromes: clinical aspects, treatment and pathophysiology.

Abstract: Cerebral creatine deficiency syndromes (CCDSs) are a group of inborn errors of creatine metabolism comprising two autosomal recessive disorders that affect the biosynthesis of creatine – i.e. arginine:glycine amidinotransferase deficiency (AGAT; MIM 602360) and guanidinoacetate methyltransferase deficiency (GAMT; MIM 601240) – and an X-linked defect that affects the creatine transporter, SLC6A8 deficiency (SLC6A8; MIM 300036). The biochemical hallmarks of these disorders include cerebral creatine deficiency as detected in vivo by 1H magnetic resonance spectroscopy (MRS) of the brain, and specific disturbances in metabolites of creatine metabolism in body fluids. In urine and plasma, abnormal guanidinoacetic acid (GAA) levels are found in AGAT deficiency (reduced GAA) and in GAMT deficiency (increased GAA). In urine of males with SLC6A8 deficiency, an increased creatine/creatinine ratio is detected. The common clinical presentation in CCDS includes mental retardation, expressive speech and language delay, autistic like behaviour and epilepsy. Treatment of the creatine biosynthesis defects has yielded clinical improvement, while for creatine transporter deficiency, successful treatment strategies still need to be discovered. CCDSs may be responsible for a considerable fraction of children and adults affected with mental retardation of unknown etiology. Thus, screening for this group of disorders should be included in the differential diagnosis of this population. In this review, also the importance of CCDSs for the unravelling of the (patho)physiology of cerebral creatine metabolism is discussed

Pathologies involving the S100 proteins and RAGE.

Abstract: The S100 proteins are exclusively expressed in vertebrates and are the largest subgroup within the superfamily of EF-hand Ca 2+ -binding proteins

Physiological Roles Of The Ca2+/Cam-Dependent Protein kinase Cascade In Health and Disease

Abstract: Numerous hormones, growth factors and physiological processes cause a rise in cytosolic Ca2+, which is translated into meaningful cellular responses by interacting with a large number of Ca2+-binding proteins. The Ca2+-binding protein that is most pervasive in mediating these responses is calmodulin (CaM), which acts as a primary receptor for Ca2+ in all eukaryotic cells. In turn, Ca2+/CaM functions as an allosteric activator of a host of enzymatic proteins including a considerable number of protein kinases. The topic of this review is to discuss the physiological roles of a sub-set of these protein kinases which can function in cells as a Ca2+/CaM-dependent kinase signaling cascade. The cascade was originally believed to consist of a CaM kinase kinase that phosphorylates and activates one of two CaM kinases, CaMKI or CaMKIV. The unusual aspect of this cascade is that both the kinase kinase and the kinase require the binding of Ca2+/CaM for activation. More recently, one of the CaM kinase kinases has been found to activate another important enzyme, the AMP-dependent protein kinase so the concept of the CaM kinase cascade must be expanded. A CaM kinase cascade is important for many normal physiological processes that when misregulated can lead to a variety of disease states. These processes include: cell proliferation and apoptosis that may conspire in the genesis of cancer; neuronal growth and function related to brain development, synaptic plasticity as well as memory formation and maintenance; proper function of the immune system including the inflammatory response, activation of T lymphocytes and hematopoietic stem cell maintenance; and the central control of energy balance that, when altered, can lead to obesity and diabetes. Although the study of the CaM-dependent kinase cascades is still in its infancy continued analysis of the pathways regulated by these Ca2+-initiated signaling cascades holds considerable promise for the future of disease-related research

The calcium-sensing receptor: physiology, pathophysiology and CaR-based therapeutics.

Abstract: The extracellular calcium (Ca2+_o)-sensing receptor (CaR) enables the parathyroid glands and other CaR-expressing cells to sense alterations in the level of Ca2+_o and to respond with changes in function that are directed at normalizing the blood calcium concentration In addition to the parathyroid gland, the kidney is a key site for Ca2+_o-sensing that enables it to make physiologically relevant alterations in divalent cation and water metabolism Several disorders of Ca2+_o-sensing arise from inherited or acquired abnormalities that “reset” the serum calcium concentration upward or downward Inactivating mutations produce a benign form of hypercalcemia when present in the heterozygous state, termed Familial Hypocalciuric Hypercalcemia (FHH), while homozygous mutations produce a much more severe hypercalcemic disorder resulting from marked hyperparathyroidism, called Neonatal Severe Hyperparathyroidism (NSHPT) Activating mutations cause a hypocalcemic syndrome of varying severity, termed autosomal dominant hypocalcemia or hypoparathyroidism Inactivating or activating antibodies directed at the CaR produce the expected hyper- or hypocalcemic syndromes, respectively “Calcimimetic” CaR activators and “calcilytic” CaR antagonists have been developed The calcimimetics are currently in use for controlling severe hyperparathyroidism in patients receiving dialysis treatment for end stage renal disease or with parathyroid cancer Calcilytics are being evaluated as a means of inducing a “pulse” in the circulating parathyroid hormone (PTH) concentration, which would mimic that resulting from injection of PTH, an established anabolic form of treatment for osteoporosis

Kinetics of peroxiredoxins and their role in the decomposition of peroxynitrite.

Abstract: Methodologies and results of studies on the kinetics of peroxiredoxins (Prx) are reviewed. Peroxiredoxins are broad-spectrum peroxidases that catalyze the reduction of H2o2, organic hydroperoxides and peroxynitrite by thiols. Their catalytic cycle starts with the oxidation of a particularly reactive cysteine residue (CP) to a sulfenic acid derivative by the peroxide substrate, the sulfenic acid then reacts with a thiol to form a disulfide, and the cycle is completed by thiol/disulfide exchange reactions that regenerate the ground-state enzyme. Depending on the subtype of peroxiredoxin, the thiol reacting with the primary oxidation product (E-SOH) may be a cysteine residue of a second subunit (typical 2-Cys Prx), a cysteine residue of the same subunit (atypical 2-Cys Prx) or reducing substrate (1-Cys Prx and at least one example of an atypical 2-Cys Prx). In a typical 2-Cys Prx the intra-subunit disulfide formation with the second “resolving” cysteine (CR) is mandatory for the reduction by the specific substrate, which is a protein characterized by a CXXC motif such as thioredoxin, tryparedoxin or AhpF. These consecutive redox reactions define the catalysis as an enzyme substitution mechanism, which is corroborated by a ping-pong pattern that is commonly observed in steady–state analyses, chemical identification of catalytic intermediates and stopped-flow analyses of partial reactions. More complex kinetic patterns are discussed in terms of cooperativity between the subunits of the oligomeric enzymes, generation of different oxidized intermediates or partial over-oxidation of CP to a sulfinic acid. Saturation kinetics is often not observed indicating that a typical complex between reduced enzyme and hydroperoxide is not formed and that, in these cases, formation of the complex between the oxidized enzyme and its reducing substrate is slower than the reaction within this complex. Working with sulphur catalysis, Prxs are usually less efficient than the heme- or selenium-containing peroxidases, but in some cases the k+1 values (bimolecular rate constant for oxidation of reduced E by ROOH) are comparable, the overall range being 2× 103-4× 107M-1 s-1 depending on the hydroperoxide and the individual Prx. For the reduction of peroxynitrite k+1 values of 1× 106 up to 7× 107 M-1s-1 have been measured. The net forward rate constants k′ +2 for the reductive part of the cycle range between 2× 104–1× 107 M-1s-1. These kinetic characteristics qualify the peroxiredoxins as moderately efficient devices to detoxify hydroperoxides, which is pivotal to organisms devoid of more efficient peroxidases, and as most relevant to the detoxification of peroxynitrite. In higher organisms, their specific role is seen in the regulation of signalling cascades that are modulated by H2o2, lipid hydroperoxides or peroxynitrite

Protein networks and complexes in photoreceptor cilia.

Abstract: Vertebrate photoreceptor cells are ciliated sensory cells specialized for single photon detection. The photoreceptor outer segment corresponds to the ciliary shaft of a prototypic cilium. In the outer segment compartment, the ciliary membrane is highly modified into membranous disks which are enveloped by the plasma membrane in rod cells. At these outer segment disks, the visual transduction cascade--a prototypical G-protein coupled receptor transduction pathway is arranged. The light sensitive outer segments are linked by the socalled connecting cilium with the inner segment, the photoreceptor compartment which contains all organelles necessary for cell metabolism. The connecting cilium correlates with the transition zone, the short junction between the basal body and the axoneme of a prototypic cilium. The connecting cilium and the calycal processes, including the periciliary ridge complex, as well as the basal body complex are in close functional association with each other. In the latter ciliary compartments, the export and import from/into the outer segment of the photoreceptor cell are controlled and regulated. In all subciliary compartments, proteins are arranged in functional multiprotein complexes. In the outer segment, signaling components are arranged into complexes which provide specificity and speed for the signaling and serve in adaptation. Centrin-G-protein complexes may regulate the light driven translocation of the visual G-protein transducin through the connecting cilium. Intraflagellar transport (IFT) complexes may serve in intersegmental exchange of molecules. The import/export of molecules is thought to be regulated by proteins arranged in networks at the basal body complex. Proteins of the interactome related to the human Usher syndrome are localized in the connecting cilium and may participate in the ciliary transport, but are also arranged at interfaces between the inner segment and the connecting cilium where they probably control the cargo handover between the transport systems of the inner segment and these of the cilium. Furthermore, USH protein complexes may further provide mechanical stabilization to membrane specializations of the calycal processes and the connecting cilium. The protein complex in which the retinitis pigmentosa GTPase regulator (RPGR) participates in the ciliary compartments also plays a key role in the function and maintenance of photoreceptor cells. It further associates through the presumed scaffolding protein RPGRIP1 with the nephrocystin protein network. Although many of these proteins have been also found in prototypic cilia or primary cilia, the arrangements of the proteins in complexes can be specific for vertebrate photoreceptor cells. Defects of proteins in these complexes lead to photoreceptor cell death and retinal degeneration, underlying syndromic and non-syndromic blindness.

SERCA pumps and human diseases.

Abstract: Sarco(endo)plasmic reticulum (SER) Ca2+ ATPases represent a highly conserved family of Ca2+ pumps which actively transport Ca2+ from the cytosol to the SER against a large concentration gradient. In humans, 3 genes (ATP2A1-3) generate multiple isoforms (SERCA1a,b, SERCA2a–c, SECA3a–f) by developmental or tissue-specific alternative splicing. These pumps differ by their regulatory and kinetic properties, allowing for optimized function in the tissue where they are expressed. They play a central role in calcium signalling through regenerating SER Ca2+ stores, maintaining appropriate Ca2+ levels in this organelle and shaping cytosolic and nuclear Ca2+ variations which govern cell response. Defects in ATP2A1 encoding SERCA1 cause recessive Brody myopathy, mutations in ATP2A2 coding for SERCA2 underlie a dominant skin disease, Darier diseaseand its clinical variants. SERCA2a expression is reduced in heart failure in human and in mice models. Gene-targeting studies in mouse confirmed the expected function of these isoforms in some cases, but also resulted in unexpected phenotypes: SERCA1 null mutants die from respiratory failure, SERCA2 heterozygous mutant mice develop skin cancer with age and SERCA3 null mice display no diabetes. These nique phenotypes have provided invaluable information on the role of these pumps in specific tissues and species, and have improved our understanding of Ca2+ regulated processes in muscles, the heart and the skin in human and in mice. Although the understanding of the pathogenesis of these diseases is still incomplete, these recent advances hold the promise of improved knowledge on the disease processes and the identification of new targets for therapeutic interventions

Functions of the histone chaperone nucleolin in diseases.

Abstract: Alteration of nuclear morphology is often used by pathologist as diagnostic marker for malignancies like cancer. In particular, the staining of cells by the silver staining methods (AgNOR) has been proved to be an important tool for predicting the clinical outcome of some cancer diseases. Two major argyrophilic proteins responsible for the strong staining of cells in interphase are the nucleophosmin (B23) and the nucleolin (C23) nucleolar proteins. Interestingly these two proteins have been described as chromatin associated proteins with histone chaperone activities and also as proteins able to regulate chromatin transcription. Nucleolin seems to be over-expressed in highly proliferative cells and is involved in many aspect of gene expression: chromatin remodeling, DNA recombination and replication, RNA transcription by RNA polymerase I and II, rRNA processing, mRNA stabilisation, cytokinesis and apoptosis. Interestingly, nucleolin is also found on the cell surface in a wide range of cancer cells, a property which is being used as a marker for the diagnosis of cancer and for the development of anti-cancer drugs to inhibit proliferation of cancer cells. In addition to its implication in cancer, nucleolin has been described not only as a marker or as a protein being involved in many diseases like viral infections, autoimmune diseases, Alzheimer's disease pathology but also in drug resistance. In this review we will focus on the chromatin associated functions of nucleolin and discuss the functions of nucleolin or its use as diagnostic marker and as a target for therapy

Calcium and Cardiomyopathies

Abstract: Regulation of Calcium (Ca) cycling by the sarcoplasmic reticulum (SR) underlies the control of cardiac contraction during excitation-contraction (E-C) coupling. Moreover, alterations in E-C coupling occurring in cardiac hypertrophy and heart failure are characterized by abnormal Ca-cycling through the SR network. A large body of evidence points to the central role of: a) SERCA and its regulator phospholamban (PLN) in the modulation of cardiac relaxation; b) calsequestrin in the regulation of SR Ca-load; and c) the ryanodine receptor (RyR) Ca-channel in the control of SR Ca-release. The levels or activity of these key Ca-handling proteins are altered in cardiomyopathies, and these changes have been linked to the deteriorated cardiac function and remodeling. Furthermore, genetic variants in these SR Ca-cycling proteins have been identified, which may predispose to heart failure or fatal arrhythmias. This chapter concentrates on the pivotal role of SR Ca-cycling proteins in health and disease with specific emphasis on their recently reported genetic modifiers

The catalytic mechanism of peroxiredoxins.

Abstract: Peroxiredoxins carry out the efficient reduction of a typically broad range of peroxide substrates through an absolutely conserved, activated cysteine residue within a highly conserved active site pocket structure. Though details of reductive recycling after cysteine sulfenic acid formation at the active site vary among members of different Prx classes, local unfolding around the active site cysteine is likely generally required in these proteins for disulfide bond formation with a second resolving cysteine and/or for access of the reductant to the oxidized active site. The conformational change associated with the catalytic cycle and the redox-dependent decamer formation occurring in at least some typical 2-Cys Prxs have interesting implications in the interplay between active site loop dynamics, oligomerization state, catalytic efficiency and propensity toward inactivation during turnover in these important antioxidant enzymes

Nutritional intervention in brain aging: reducing the effects of inflammation and oxidative stress.

Abstract: It is estimated that by the year 2050 the elderly (aged 65 or older) population will double the population of children (aged 0-14) for the first time in history. The expansion of the elderly population has already taken a toll on health care systems. In order to alleviate the health care costs and increase the quality of living in the aging population, it is crucial to explore methods that may retard or reverse the deleterious effects of aging. Inflammation and oxidative stress play important roles in brain aging. Inflammatory markers, as well as cellular and molecular oxidative damage, increase during normal brain aging. This increase is accompanied by the concomitant decline in cognitive and motor performance in the elderly population, even in the absence of neurodegenerative diseases. Epidemiological studies have shown that consumption of diets rich in antioxidant and anti-inflammatory agents, such as those found in fruits and vegetables, may lower the risk of developing age-related neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Research from our laboratory suggests that dietary supplementation with fruit or vegetable extracts can decrease the age-enhanced vulnerability to oxidative stress and inflammation. Additional research suggests that the polyphenolic compounds found in fruits such as blueberries may exert their beneficial effects through signal transduction and neuronal communication. Thus, nutritional intervention may exert therapeutic protection against age-related deficits and neurodegenerative diseases.

Peroxiredoxins in bacterial antioxidant defense.

Abstract: Peroxiredoxins constitute an important component of the bacterial defense against toxic peroxides. These enzymes use reactive cysteine thiols to reduce peroxides with electrons ultimately derived from reduced pyridine dinucleotides. Studies examining the regulation and physiological roles of AhpC, Tpx, Ohr and OsmC reveal the multi-layered nature of bacterial peroxide defense. AhpC is localized in the cytoplasm and has a wide substrate range that includesH2O2, organic peroxides and peroxynitrite. This enzyme functions in both the control of endogenous peroxides, as well as in the inducible defense response to exogenous peroxides or general stresses. Ohr, OsmC and Tpx are organic peroxide specific. Tpx is localized to the periplasm and can be involved in either constitutive peroxide defense or participate in oxidative stress inducible responses depending on the organism. Ohr is an organic peroxide specific defense system that is under the control of the organic peroxide sensing repressor OhrR. In some organisms Ohr homologs are regulated in response to general stress. Clear evidence indicates that AhpC, Tpx and Ohr are involved in virulence. The role of OsmC is less clear. Regulation of OsmC expression is not oxidative stress inducible, but is controlled by multiple general stress responsive regulators

Obesity, inflammation, and vascular disease: the role of the adipose tissue as an endocrine organ

Abstract: Insulin resistance, both in nondiabetic and diabetic subjects, is frequently associated with obesity, particularly with an excess of central fat. With the growing prevalence of obesity, scientific interest in the biology of adipose tissue has been extended to the secretory products of adipocytes, since they are increasingly shown to influence several aspects in the pathogenesis of obesity-related diseases Until relatively recently, the role of fat itself in the development of obesity and its consequences was considered to be a passive one; adipocytes were considered to be little more than storage cells for fat. It is now clear that, in addition to storing calories as triglycerides, they also secrete a large variety of proteins, including cytokines, chemokines and hormone-like factors, such as leptin, adiponectin and resistin. This production of pro-atherogenic chemokines by adipose tissue is of particular interest since their local secretion, e.g. by perivascular adipose depots, may provide a novel mechanistic link between obesity and the associated vascular complications. Recent research has revealed many functions of adipocytokines extending far beyond metabolism, such as immunity, cancer and bone formation. This remarkable understanding is allowing us to more clearly define the role that adipocytes play in health and in obesity and how inflammatory mediators act as signaling molecules in this process. Moreover, on a molecular level, we are beginning to comprehend how such variables as hormonal control, exercise, food intake, and genetic variation interact and result in a given phenotype, and how pharmacological intervention may modulate adipose tissue biology.

Role of histone phosphorylation in chromatin dynamics and its implications in diseases

Abstract: In eukaryotic cells, relaxed interphase chromatin undergoes pronounced changes resulting in formation of highly condensed mitotic chromosomes. Moreover, chromatin condensation is particularly evident during mitosis and apoptotic cell death, whereas chromatin relaxation is necessary for replication, repair, recombination and transcription. The post-translational modifications of histone tails such as reversible acetylation, phosphorylation and methylation play a critical role in dynamic condensation/relaxation that occurs during the cell cycle. Histone phosphorylation is believed to play a direct role in mitosis, cell death, repair, replication and recombination. However, definitive roles for this modification in these processes have not yet been elucidated. In this review, we discuss recent progress in studies of histone phosphorylation.

Evolution of the peroxiredoxins.

Abstract: Peroxiredoxins compose a superfamily of peroxidases ubiquitously found throughout evolution in prokaryotes, archaea and eukaryotes. These enzymes contain a conserved catalytic peroxidatic cysteine (Cp) in the N-terminal region of the protein. The residues surrounding Cp and the catalytic site appear also to be well conserved. Peroxiredoxins can be classified either into three subfamilies according to their catalytic mechanism or into five subfamilies according to sequence homology. Notably, the number of peroxiredoxin genes increased during evolution. In eukaryotes, the higher number of genes coding for peroxiredoxin family members is due to the existence of different isoforms targeted to different subcellular compartments but is probably due also to the acquisition of new functions. Indeed, it has been postulated that the antioxidant protective role of peroxiredoxins, which is particularly critical in prokaryotes, in yeasts and in parasitic eukaryotes, may have evolved to a modulatory role in hydrogen peroxide signaling in plants and animals.

Structural Survey of the Peroxiredoxins

Abstract: Peroxiredoxins (Prxs) are ubiquitous proteins that use an active site Cys residue to reduce hydroperoxides. Structural studies since the first Prx structure was determined in 1998 have produced 35 crystal structures of wild type and mutant Prxs with at least one representative structure from each of the five major evolutionary subfamilies of Prxs. These structures have yielded a great deal of knowledge about Prx structure and structure-function relations, revealing fascinating variations in quaternary structure and details of the fully-folded and locally-unfolded conformations that are involved in the catalytic cycle of all Prxs

Proteomic analysis of secreted exosomes.

Abstract: This chapter focuses on the contribution of proteomic analysis to the understanding of the process of exosome secretion and the mechanism and function of exosomes. It also describes the potential of exosome proteomic analysis to aid in the development of exosomes for therapeutic use.

Role of COX-2 in inflammatory and degenerative brain diseases.

Abstract: In the last decade, the potential role of cyclooxygenase-2 (COX-2) and prostaglandins (PGs) in brain diseases has been extensively studied. COX-2 over-expression has been associated with neurotoxiticy in acute conditions, such as hypoxia/ischemia and seizures, as well as in inflammatory chronic diseases, including Creutzfeldt-Jakob disease (CJD) and Alzheimer's disease (AD). However, the role played by COX-2 in neurodegenerative diseases is still controversial and further clinical and experimental studies are warranted. In addition, the emerging role of COX-2 in behavioural and cognitive functions strongly indicates that studies aimed at improving our knowledge of the physiological role of COX-2 in the central nervous system are crucial to fully understand the pros and cons of its manipulation in disabling neurological diseases

Calcium signalling and cancer cell growth.

Abstract: Cancer is caused by defects in the mechanisms underlying cell proliferation and cell death. Calcium ions are central to both phenomena, serving as major signalling agents with spatial localization, magnitude and temporal characteristics of calcium signals ultimately determining cell’s fate. There are four primary compartments: extracellular space, cytoplasm, endoplasmic reticulum and mitochondria that participate in the cellular Ca2+ circulation. They are separated by own membranes incorporating divers Ca2+-handling proteins whose concerted action provides for Ca2+ signals with the spatial and temporal characteristics necessary to account for specific cellular response. The transformation of a normal cell into a cancer cell is associated with a major re-arrangement of Ca2+ pumps, Na/Ca exchangers and Ca2+ channels, which leads to the enhanced proliferation and impaired ability to die. In the present chapter we examine what changes in Ca2+ signalling and the mechanisms that support it underlie the passage from normal to pathological cell growth and death control. Understanding this changes and identifying molecular players involved provides new prospects for cancers treatment

Cancer chemoprevention by cyclooxygenase 2 (COX-2) blockade: results of case control studies.

Abstract: Significant use of selective cyclooxygenase-2 (COX-2) blocking agents prescribed for the treatment of arthritis during 1999 to 2005 facilitates epidemiologic investigations to illuminate their chemopreventive effects against human cancer We therefore conducted a set of case control studies of selective COX-2 blocking agents to determine their chemopreventive potential for the four major cancers: breast, prostate, colon, and lung Newly diagnosed cases (323 breast cancer patients, 229 prostate cancer patients, 326 colon cancer patients, and 486 lung cancer patients) were ascertained during 2002 to September 30, 2004, at The James Cancer Hospital and Solove Research Institute, The Ohio State University Medical Center, Columbus, Ohio All cases of invasive cancer were confirmed by examination of the pathology report Healthy controls without cancer were ascertained from hospital screening clinics during the same time period Controls were frequency matched at a rate of 2:1 to the cases by age, gender, and county of residence We collected information on type, frequency, and duration of use of selective COX-2 inhibitors and nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) Other potentially important risk factors (smoking, drinking, body mass, medical history, blood pressure and cholesterol medications, family history of cancer, occupational history, and reproductive history for women) were also recorded for each subject Estimates of odds ratios were obtained with adjustment for age and other potential confounders using logistic regression analysis Use of selective COX-2 inhibitors resulted in a significant risk reduction for each type of cancer (71% for breast cancer, 55% for prostate cancer, 70% for colon cancer, and 79% for lung cancer) and an overall 68% risk reduction for all four cancers This investigation demonstrates that COX-2 blocking agents have strong potential for the chemoprevention of cancers of the breast, prostate, colon and lung

Functional insights into the creatine transporter.

Abstract: Creatine and phosphocreatine provide an intracellular, high-energy phosphate buffering system, essential to maintain ATP levels in tissues with high energy demands. A specific plasma membrane creatine transporter (CRT) is required for the cellular uptake of creatine. This transporter is related to the \UPgamma -aminobutyric acid (GAT) and norepinephrine (NET) transporters and is part of a large gene family of Na+- and Cl--dependent neurotransmitter transporters, now known as solute carrier family 6 (SLC6). CRT is essential for normal brain function as mutations in the CRT gene (SLC6A8) result in X-linked mental retardation, associated with the almost complete lack of creatine in the brain, severe speech and language delay, epilepsy, and autistic behaviour. Insight into the structure and function of the CRT has come from studies of creatine transport by tissues and cells, in vitro studies of CRT mutations, identification of mutations associated with CRT deficiency, and from the recent high resolution structure of a prokaryotic homologue of the SLC6 transporters. CRT antibodies have been developed enabling the localization of creatine uptake sites in the brain, retina, muscle and other tissues. These tools in conjunction with the use of appropriate cell models should allow further progress in our knowledge on the regulation and cellular trafficking of the CRT. Development of suitable mouse models may allow improved understanding of the importance of the CRT for normal brain function and how the transporter is regulated in vivo

The peroxiredoxin repair proteins.

Abstract: Sulfiredoxin and sestrin are cysteine sulfinic acid reductases that selectively reduce or repair the hyperoxidized forms of typical 2-Cys peroxiredoxins within eukaryotes. As such these enzymes play key roles in the modulation of peroxide-mediated cell signaling and cellular defense mechanisms. The unique structure of sulfiredoxin facilitates access to the peroxiredoxin active site and novel sulfur chemistry

Histone acetylation and methylation: combinatorial players for transcriptional regulation.

Abstract: Post-synthetic modification of histone proteins in chromatin architecture plays a central role in the epigenetic regulation of transcription. Histone acetylation and methylation are the two major modifications that function as a specific transcription regulator in response to various cellular signals. Albeit the mechanism of action of these modifications in transcription is not well understood, recent discovery of histone acetyltransferase (HAT) and methyltransferase (HMT) activities within transcriptional regulators has an important implication for histone modification to be a key player for the precise regulation of transcription processes. Here, we discuss recent advances made on histone acetylation and methylation as a fundamental process to modulate gene transcription, with a particular emphasis on their combinatorial effects in transcriptional control.

The creatine kinase phosphotransfer network: thermodynamic and kinetic considerations, the impact of the mitochondrial outer membrane and modelling approaches.

Abstract: In this review, we summarize the main structural and functional data on the role of the phosphocreatine (PCr) -- creatine kinase (CK) pathway for compartmentalized energy transfer in cardiac cells. Mitochondrial creatine kinase, MtCK, fixed by cardiolipin molecules in the vicinity of the adenine nucleotide translocator, is a key enzyme in this pathway. Direct transfer of ATP and ADP between these proteins has been revealed both in experimental studies on the kinetics of the regulation of mitochondrial respiration and by mathematical modelling as a main mechanism of functional coupling of PCr production to oxidative phosphorylation. In cells in vivo or in permeabilized cells in situ, this coupling is reinforced by limited permeability of the outer membrane of the mitochondria for adenine nucleotides due to the contacts with cytoskeletal proteins. Due to these mechanisms, at least 80% of total energy is exported from mitochondria by PCr molecules. Mathematical modelling of intracellular diffusion and energy transfer shows that the main function of the PCr -- CK pathway is to connect different pools (compartments) of ATP and, by this way, to overcome the local restrictions and diffusion limitation of adenine nucleotides due to the high degree of structural organization of cardiac cells

Calcium signalling and calcium transport in bone disease.

Abstract: Calcium transport and calcium signalling mechanisms in bone cells have, in many cases, been discovered by study of diseases with disordered bone metabolism. Calcium matrix deposition is driven primarily by phosphate production, and disorders in bone deposition include abnormalities in membrane phosphate transport such as in chondrocalcinosis, and defects in phosphate-producing enzymes such as in hypophosphatasia. Matrix removal is driven by acidification, which dissolves the mineral. Disorders in calcium removal from bone matrix by osteoclasts cause osteopetrosis. On the other hand, although bone is central to management of extracellular calcium, bone is not a major calcium sensing organ, although calcium sensing proteins are expressed in both osteoblasts and osteoclasts. Intracellular calcium signals are involved in secondary control including cellular motility and survival, but the relationship of these findings to specific diseases is not clear. Intracellular calcium signals may regulate the balance of cell survival versus proliferation or anabolic functional response as part of signalling cascades that integrate the response to primary signals via cell stretch, estrogen, tyrosine kinase, and tumor necrosis factor receptors

The trypanothione system.

Abstract: Trypanosomes and Leishmania, the causative agents of severe tropical diseases, employ 2-Cys-peroxiredoxins together with cysteine-homologues of glutathione peroxidases and ascorbate-dependent peroxidases for the detoxification of hydroperoxides. All three types of peroxidases gain their reducing equivalents from the parasite-specific dithiol trypanothione [bis(glutathionyl)spermidine]. Based on their primary structure and cellular localization, the trypanosomatid 2-Cys-peroxiredoxins are subdivided into two families that occur in the mitochondrion and cytosol of the parasites. In Trypanosoma brucei, the cytosolic 2-Cys-peroxiredoxin, as well as the glutathione peroxidase-type enzyme, is essential for cell viability. Despite overlapping substrate specificities and subcellular localizations, the two types of peroxidases can obviously not substitute for each other which suggests distinct cell-physiological roles

Calcium and Cell Death

Abstract: Calcium signalling system controls majority of cellular reactions. Calcium signals occurring within tightly regulated temporal and spatial domains, govern a host of Ca2+-dependent enzymes, which in turn determine specified cellular responses. Generation of Ca2+ signals is achieved through coordinated activity of several families of Ca2+ channels and transporters differentially distributed between intracellular compartments. Cell damage induced by environmental insults or by overstimulation of physiological pathways results in pathological Ca2+ signals, which trigger necrotic or apoptotic cellular death