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My SC2 comic!
  PokerDoc88, Mar 23 2009

I spent way too long doing this...I thought it'd take 2 hours but that bloated out to almost 4x as long...

Anyway, hope you enjoy!




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Thesis STINKS
  PokerDoc88, Mar 11 2009

Okay, so I'm doing research and have been busy writing my thesis. All I can say is, writing thesis = massive tilt. Literature review is a waste of time, it teaches nothing and paraphrases every bloody piece of research done in a field already, just diluting the pool of knowledge with "phantom" knowledge (IDK i just made that up, I hope you know what I mean) making it more difficult for a simple meme to be passed from person to person (confusion inevitably arises when some noob student like me misinterprets a piece of information, paraphrases it incorrectly, and then unknowingly creates a "false" piece of information which filters through to every person who reads my particular thesis). Anyway, here is my thesis copy+pasted from word in all its glory for those masochists who actually want to read this (obviously it's a work in progress).
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Part 1: Cardiovascular disease overview:
1. Types of cardiovascular disease:
Cardiovascular diseases (CVD’s) are diseases of both the heart and circulation. CVD’s include coronary heart disease (CHD), cerebrovascular disease/stroke (CVD), peripheral arterial disease (PAD), rheumatic heart disease, deep vein thrombosis (DVT), pulmonary embolism, and congenital heart dis
ease.[1]. CHD and CVD are diseases of the blood vessels supplying the heart-muscle and brain respectively, while PAD is a generalised term for disease of the blood vessels supplying the limbs. These 3 forms of CVD occur when the blood vessels supplying the respective organs become occluded, causing reduced blood flow. The most common cause of restricted blood flow is atheroma, an accumulation of lipid within the vessel intima, which stenoses the vessel lumen. I must add some info here about rupturing plaques causing thromboembolism
2. Burden:
CVDs are the leading cause of mortality globally. In 2005, it is estimated that 17.5 million people died from CVDs; this figure represents 30% of all deaths globally[1]. Of these 17.5million CVD related deaths, 7.6 million were due to CHD and 5.7 million were due to stroke[1]. It is projected that in 2015 CVD will remain the leading cause of global death, reaching a rate of 20million per annum[1]. 80% of CVD deaths occur in low and middle income countries, with an almost equal prevalence in men and women[1]. In high-income countries, lower socioeconomic groups have a relatively higher risk of diseases and mortality; a similar pattern is beginning to develop as the CVD epidemic grows in low and middle income countries[1].
CVDs limit the working capacity and savings of affected individuals, many of whom are middle aged[1]. The loss of income and healthcare payments place a developmental burden on the communities and governments of those people affected[1]. WHO data estimates that during the next 10 years (2006-2015), China will spend $558 billion of national income to manage heart disease, stroke and diabetes[1].

Part 2: Atherosclerosis
1. Overview:
The word atherosclerosis is defined by its two component words: athero which in Greek means gruel, and sclerosis which means hardening. These two words reflect the pathological characteristics of an atherosclerotic plaque; the ‘gruel’ is the necrotic debris at the center of the plaque, and the ‘hardening’ describes the fibrotic luminal border of the plaque[2].
Atherosclerotic lesions begin as fatty streaks (type I and II), which are determined as an increase in the number of macrophages present in the intima, possessing a cytosol filled with lipid droplets[3]. These streaks are clinically benign, but they are the precursors to clinically noticeable, relevant lesions[3]. One hypothesis states that platelet or fibrin deposition on the intima surface could be the initial trigger of atherogenesis[4], [5]. These fatty streaks are common in infants and young children [6], and have been described as purely inflammatory in nature, consisting of only monocyte-derived macrophages and T lymphocytes [7]. As the lesion progresses to type III, accumulation of cholesterol esters in the extracellular space occurs[2]. The development of the lesion to later, clinically relevant types is clearly described by Stary; in a type IV lesion, calcium deposits may occur within the cellular organelles and the extracellular matrix, and extracellular cholesterol crystals also appear[8]. The layer of tissue overlying the core is composed of proteoglycan-rich intercellular matrix, smooth muscle cells, macrophages, macrophage foam cells (macrophages with large intracellular lipid droplets), and lymphocytes. This tissue layer readily allows the development of fibrous connective tissue, surface disruption, and thrombi, although none of these features are characteristic of a type IV lesion[8]. Further deposition of fibrous connective tissue layers, which can occur over a period of years to decades after the formation of the type IV lesion, marks the progression to a type V lesion, known as fibroatheroma. It is believed that the interactions of macrophages, smooth muscle cells, and T cells is responsible for the fibrous tissue remodeling that occurs as lesions progress from type IV to type V[9].
The layers of fibrous tissue are laid down by intimal smooth muscle cells, which themselves are increased in number[8]. These intimal smooth muscle cells are in a “synthetic state”, and contain increased levels of rough endoplasmic reticulm, Golgi apparatus, and free ribosomes[9]. These cells function differently to smooth muscle cells present in the media, which are in a “contractile state”; the media smooth muscle cells have relatively more myofilaments and are responsible for vessel contractility [10]. Type V lesions can be subclassified into Va, Vb, and Vc lesions [9]. Va lesions posses a mixed lipid and fibrous tissue composition; Vb lesions display prominent calcification [9] in the form of hydroxyapatite crystals[11] , while Vc lesions are predominantly fibrous in nature [12].
If a lesion (typically type IV and V) develops surface disruptions, hematomas, or thrombotic material, then it is classified as a type VI lesion. Thrombotic deposits, usually composed of only fibrin, may occur on type VI lesions. This thrombogenic material complicates the development of the atheroma, sometimes acutely accelerating the growth of the lesion and resulting in sudden vessel stenosis [13]. Histologic evidence suggests that these events responsible for sudden progression of lesion development occur at irregular intervals of months to years, how ever rapid succession thrombotic layer deposition can lead to fatal ischemia in a period of hours, days, or weeks [13].
Type VIII lesions are primarily composed of fibrous tissue, which may be hyalinised; these lesions lack a lipid core, and sometimes are devoid of lipid material at any site[8]. Regression of lipid cores, extension of the fibrous material to adjacent fibroatheroma, and organization of thrombotic material may all lead to the development of a type VIII lesion [13].
Atheroma may also be described as being either ‘stable’ or ‘vulnerable’[14]. Vulnerable plaques possess a well-developed lipid core with a thin fibrous cap, which separates thrombogenic material and macrophages from the blood[3]. Vulnerable plaques are clinically significant, even if they are not occluding the arterial lumen, because complications such as thromboembolism due to plaque rupture may suddenly occur[15]. Stable plaques have a relatively thickened fibrous cap, separating the lipid core from the blood [3].

Sites of occurance:
Atherosclerotic lesions begin their formation in the intima of susceptible blood vessels[3], however as the lesion progresses to later stages, media involvement may occur[2]. These lesions occur primarily in large and medium-sized arteries, both elastic and muscular in nature [16].
Risk factors:
Atherosclerosis is currently recognized as a multifactorial disease, caused by the effects of multiple risk factors acting in a genetically susceptible individual [16]. These risk factors can broadly be classified as being either modifiable or unmodifiable [17].
Modifiable risk factors:
Plasma concentrations of cholesterol, particularly low-density lipoprotein (LDL) cholesterol, are a major risk factor for atherosclerosis[16]. It has been recognized however that high levels of high-density lipoprotein (HDL) plays a protective role in atherogenesis [18]. LDL can be chemically modified by oxidation, glycosylisation, aggregation, association with proteoglycans or incorporation into immune complexes [19]. These chemical changes to LDL can result in injury to the endothelium and underlying smooth muscle [20].
If LDL particles remain stagnant in an artery, they can undergo progressive oxidation and become endocytosed by scavenger macrophages[19]. Once inside the cell, these LDL particles form into lipid peroxides, which facilitate the accumulation of cholesterol esters, eventually giving rise to foam cells [16]. These foam cells remove and sequester the modified LDL particles, as part of an initial protective mechanism in the inflammatory response[16]. Antioxidants such as Vitamin E also assist in this protective response by reducing levels of free-radicals formed by the modified LDL [21]. The LDL also acts as a chemotactic agent for other monocytes, and increases expression of genes coding the production of macrophage-colony stimulating factor(MCSF) [22]and monocyte chemotactic protein [23]from endothelial cells.
The inflammatory response also alters the movement of lipoprotein within the artery by increasing the ability of LDL to bind to endothelium and smooth muscle through increasing the expression of the LDL receptor gene[24, 25]. This upregulation of LDL receptors is believed to be caused by the upregulation of inflammatory cytokines such as tumor necrosis factor alpha (TNF-α, interleukin-1 (IL-1), and MCSF[24, 25]. The intracellular events triggered by LDL binding its scavenger receptor in vitro[25] include the induction of urokinase[26] and IL-1, as well as other inflammatory cytokines[27-29], which serves to maintain the cycle of inflammation and lipoprotein modification[16].
Even though pharmacologic approaches have been implemented to lower plasma cholesterol levels[30], CVD persists as the primary cause of death throughout Europe, Asia, and the United States of America [31], [32].

Homocysteine (HCY) concentrations in plasma have been associated with advanced atherosclerotic lesions[16]. HCY derived from the intracellular metabolism of methionine[33]. After being processed, it exits into the plasma where it is largely present bound to proteins in oxidized forms such as homocystine and cysteine-HCY disulfide[33]. In patients with CHD, CVD and PAD, HCY levels are elevated in 15-40% of cases [33]. The observation of high HCY was first noted in autopsy findings of patients exhibiting advanced atherosclerosis as well as homogenous defects in enzymes required for the metabolism of HCY, such as cystathionine beta-synthase and methylenetetrahydrofolate reductase[33-37]. Patients who possess these enzyme defects develop severe atherosclerosis early in childhood, and often suffer from myocardial infarction at the age of 20 years [33, 37]. Studies have shown that homocysteine is toxic to endothelium [38], is prothrombogenic [39], increases collagen production[40], and decreases the levels of nitric oxide available[41]. Elevated levels of HCY can be returned to normal levels with folic acid, occasionally requiring supplementation with pyridoxine, vitamin B12, choline or betaine [33]. Research is currently underway to determine whether folic acid treatment may cause regression of atherosclerotic lesions [42].
A recent study has shown high levels of HCY have been observed in hypertensive children[43].

Oxidative stress:
Oxidised LDL is observed in human atherosclerotic lesions[44]. In studies of animals with hypercholesterolemia, antioxidants have been shown to reduce the size of atherosclerotic lesions [45] and fatty streaks in nonhuman primates [46]. These observations suggest that antioxidants may possess an anti-inflammatory effect in humans [47]. Ex-vivo studies have shown that plasma levels of antioxidants such as vitamin E increase the resistance of LDL particles being oxidized[48], however other antioxidants such as beta carotene have displayed no such effect [48] [49].
Response-to-injury hypothesis:
The initial step in this proposed mechanism of atherosclerotic lesion development is endothelial dysfunction [50]. It is suggested that each characteristic atherosclerotic lesion is representative of a chronic inflammatory process within the artery [50]. Various factors have been suggested to cause endothelial dysfunction or denudation, including elevated and modified plasma LDL; free radicals due to cigarette smoking, hypertension and diabetes mellitus; genetic alterations; elevated plasma homocysteine concentrations; infections microorganisms such as herpesviruses and Chlamydia pneumonia[50]. The resulting injury induces homeostatic changes to the endothelium, such as increased adhesiveness for platelets and leukocytes, increased permeability, and altered function to display procoagulant instead of anticoagulative properties[50]. The endothelium is also stimulated to release vasoactive molecules, cytokines, and growth factors which causes a migration of leucocytes and proliferation of smooth muscle cells [50]. If this response persists because the compensatory response is unable to remove the harmful stimulus, over time “remodeling” occurs, which causes a thickened artery wall. The vessel can initially dilate gradually so that up to a certain point, the lumen remains unaltered [51]
An increase in the numbers of macrophages and lymphocytes present at the lesion site occurs. Activation of these cells triggers them to release cytokines, chemokines, hydrolytic enzymes, and growth factors [52] which produces local damage to the endothelium and eventually leads to focal necrosis [53]. Cycles of this process whereby mononuclear cells migrate to the lesion site and release their chemical mediators, triggers local proliferation of smooth muscle cells and formation of fibrous tissue which enlarges and restructures the lesion [16]. This results in a thickened lesion with a fibrous cap overlying a core comprised of lipid and necrotic material, which restricts the vessels’ capacity to dilate, obstructing blood-flow [16].
Autoimmune hypothesis:
It has been observed that atherosclerotic lesions begin when LDL molecules become trapped within sub-endothelial extracellular matrix of the arterial wall, causing the appearance of a “fatty streak” [54]. Already at this pre-plaque stage of lesion development, lymphocytic T-cells can be observed[55]. Later on, a large subendothelial infiltration of mononuclear cells occurs, consisting mainly of CD4+ Th1 lymphocytes (but also some CD8+ Th1 lymphocytes) as well as macrophages, monocytes, and mast cells[56-58]. Also present in the sub-endothelial space are antibody-antigen complexes and complement[56]. These observations suggest that the early stage of atherosclerosis is mediated by an immunological reaction, possibly caused by local autoantigens [55, 59].
ER Stress:
The endoplasmic reticulum (ER) is a cellular organelle which serves as the site of membrane and secretory protein synthesis and folding, synthesis of lipids and sterols, and storage of free Ca2+ [60]. The ER protein output represents a large portion of total mammalian cell protein output, hence the trafficking of protein through the ER requires careful monitoring for abnormalities including protein misfolding and accumulation [60]. Various stressors can lead to an imbalance in the requirement of protein folding and the ER capacity to fold these proteins, hence causing ER stress [60]. These stresses can be either physiological, such as an increased demand for protein secretion, or pathological such as an accumulation of unfolded mutant proteins [60].
The unfolded protein response (UPR) was first investigated upon the discovery of a set of genes that were upregulated in glucose-deprived fibroblasts[61]. The proteins resulting from these genes’ translation were named glucose-regulated proteins (GRPs)[62, 63]. The first discovered and most abundant protein in this family, GRP78 (also known as BiP) was investigated by studying incomplete immunoglobulin intermediates[64]. Another studied GRP, GRP94, was first observed in an experiment involving over-expression of an unfolded mutant influenza hemaglutinin[65]. These GRPs possess molecular chaperone functions within the ER[66].
Since study into the UPR began, several components of the pathway have been discovered including an ER transmembrane bifunctional serine/threonine protein kinase/endoribonuclease (Ire1p/Ern1p), a UPR transducer which induces BiP and other ER chaperones in yeast[67, 68]. HAC1 mRNA is the only discovered substrate to possess Ire1p RNase activity[66]. Unfolded proteins present at the ER cause Ire1p to dimerise, which results in an autophosphorylation of Ire1p; Ire1p then removes and splices a 252-base intron from HAC1 mRNA[69-71]. A basic leucine zipper (b-ZIP) transcription factor is encoded by the spliced HAC1, which initiates approximately 381 UPR genes within yeast[72].
The properties of the UPR in yeast has been conserved across all eukaryotic cells; this response has also evolved additional sensory mechanisms to generate various altered responses[66]. The Irep1 counterpart in mammals has 2 isoforms; IRE1α and IRE1β66]. IRE1α is expressed in most cells and tissues throughout the mammal, however IRE1β is restricted to intestinal epithelial tissues[73, 74]. There exists two additional stress sensors within the mammalian UPR response: PKR-like ER-associated kinase (PERK) and activating transcription factor 6 (ATF6)[66]. Both PERK and AFT6 are expressed constitutively throughout all cells[66]. PERK possess a functionally similar luminal domain to IRE1α for sensing ER stress, and a cytosolic eukaryotic translation-initiation factor 2α domain[75]. ATF6 also has a luminal domain for sensing ER stress, and is itself a transcription factor for b-ZIP[76].
During times of non-stress, GRP78 binds to and inhibits IRE1, PERK, and ATF6 to prevent their UPR signalling[66]. If the ER becomes overloaded with freshly synthesised protein or accumulations of unfolded proteins, GRP78 binds to these unfolded proteins in the ER, preventing their transport to the cis-Golgi[66]. As more unfolded proteins become bound by GRP78, the pool of GRP78 inhibiting IRE1, ATF6 and PERK becomes squandered; this allows IRE1 and PERK to homodimerise, autophosphorylate, and become active[77, 78], and ATF6 to translocate to the Golgi compartment where it is cleaved to become cytosolically active and migrate to the nucleus[79]. Despite the fact that GRP78 regulates the three UPR transducers within most cells, in certain cells sometimes a specific stressor will activate only one or two of the UPR pathways[66]. For instance, the IRE1α-mediated UPR pathway is activated during B cell differentiation and the PERK pathway is inactive[78, 80], while in pancreatic β cells, glucose limitation seems to selectively activate PERK in preference of IRE1 (Scheuner and Kaufman, unpublished observation)[66].
Some of the mechanisms activated by cells during the UPR in order to survive accumulations of unfolded proteins in the ER lumen include reducing the amount of new protein translocated into the ER, reducing DNA translation initiation, increasing the degradation rate of misfolded proteins, increasing the protein-folding capacity of the ER, and retrotranslocating proteins out of the ER[66]. The promoter regions of many mammalian UPR-inducible genes including GRP78, GRP94n and calreticulin possess an ER stress response element (ERSE), which requires activation to allow transcription to occur[81]. Two UPR-specific transcription factors capable of binding the ERSE have been discovered to exist in mammals: X-box DNA binding protein 1 (XBP1) and ATF6[81]. XBP1 is a homologue of yeast HAC1 which acts as a substrate for mammalian IRE1 RNase[82-84]. Two forms of ATF6 exist, ATF6α which is 90kDa, and ATF6β (also known as CREB-RP) which is 110kDa[76]. Both ATF6α and ATF6β require that transcription factor NF-Y be present in order to bind to the ERSE[85, 86]. When unbound from GRP78, ATF6 tranlocates to the Golgi and becomes cleaved by site-1 and site-2 protease (S1P and S2P), forming a 50kD cytosolic b-ZIP-containing fragment which travels to the nucleus to activate transcription[79]. S1P and S2P are the same proteases responsible for cleaving ER-associated transmembrane sterol-response element binding protein (SREBP) in an environment deprived of cholesterol[87]. The genes activated by ATF6 include ones responsible for coding ER-resident molecular chaperones and folding enzymes, while XBP1 activated genes code for a subset of ER resident chaperone genes required for protein folding, maturation, and destruction[88]. Previously it was believed that XBP1 mRNA was induced by ATF6 for the purpose of providing more substrate for IRE1 to splice[82, 85, 89]. However, recent studies suggest that XBP1 and ATF6 operate largely in parallel pathways and may be capable of interacting with one another during periods of ER stress[89, 90].
One of the immediate actions of PERK in mammalian cells during times of ER stress is to phosphorylate eukaryotic translation initiation factor eIF2α at Ser51[91, 92], resulting in protein synthesis inhibition[93]. Phosphorylation of eIF2α prevents the formation of the ternary translation initiation complex eIF2/GTP/Met-tRNAiMet, which has a generalized effect of inhibiting translation[66]. An experiment involving murine cells with a mutated residue at Ser51 on elF2α or a PERK deletion demonstrated an inability to attenuate protein synthesis during ER stress, resulting in cell death[94]. While the generalized response of phosphorylated eIF2α is to prevent over-all mRNA translation, a selective subset of mRNAs coding proteins capable of dealing with ER stress are stimulated[66]. One of the mRNAs promoted for translation by active eIF2α is activating transcription factor 4 (ATF4)[95]. ATF4 is capable of activating genes responsible for amino acid metabolism and transport, ER stress-induced apoptosis, and redox reactions[96, 97].
The ER requires energy in order to properly fold protein in its oxidizing environment[97]. During non-stress conditions, freshly synthesised proteins are present as unfolded structures at the beginning of the protein-folding pathway[66]. If an ER stress condition occurs, such as shortage of energy supply, then many of the unfolded protein intermediates become trapped in low-energy states and accumulate[66]. These protein intermediates become sequestered by GRP78, calnexin, and calreticulin[66], before being retrotranslocated to the cytoplasm and destroyed by proteasomes[98]. This process is termed ER-associated degradation (ERAD), and is controllable by the UPR[66]. If the ERAD process is insufficient in removing enough protein accumulations from the ER, then the UPR becomes activated and genes coding products to upregulate ERAD become transcribed via the IRE1-XBP1 pathway[66, 72]. It has been shown in mammalian cells that transcription of the gene encoding ER degradation-enhancing a1, 2-mannosidase-like protein (EDEM), an essential ERAD component for degrading misfolded ER glycoproteins, depends entirely on the IRE1-XBP1 signaling pathway[99-102]. If accumulations of misfolded proteins persist in the ER, prolonged UPR activation will result in programmed cell death[66].
Knowledge of three proapoptotic pathways currently exists resulting from ER stress[66]. These three pathways are regulated by IRE1, caspase-12, and PERK/CHOP respectively[66]. During ER stress, active IRE1 can interact with c-Jun-N-terminal inhibitory kinase(JIK) to recruit TRAF2 to the ER membrane[103, 104]. TRAF2 activates apoptosis-signaling kinase 1, (ASK1, also known as MAPKKK)[105], which then leads to JNK protein kinase and mitochondria/Apaf1-dependent caspase activation[103, 105, 106]. Caspase-12, an ER-associated proximal effector of the caspase activation cascade, activates caspase-9[66]. Caspase-9 then leads to caspase-3 activation, which leads to cell apoptosis[107, 108]. The ER-stress induced death-signaling pathway regulated by PERK involves activating transcription of genes possessing proapoptotic functions[66]. Activation of PERK leads to translation of the transcription factor ATF4, which itself promotes transcription of CHOP, (also known as GAD 153) a transcription factor that potentiates apoptosis


Figure:
1. GRP78 inhibits PERK during normal cellular conditions
2. During ER stress, accumulations of unfolded protein in the ER lumen cause GRP78 to unbind PERK. PERK homodimerises and autophosphorylates. It then phosphorylates elF2α.
3. eLF2α acts to inhibit general mRNA translation initiation, but stimulates mRNA translation of mRNAs involved in UPR such as GCN4 and ATF4[66].
HSP70:
Heat shock proteins such as HSP70 and heme-oxygenase-1 (HO-1) act as cellular chaperones which bind to unfolded or improperly folded proteins to allow proper folding to occur [109], and to prevent intracellular protein accumulation and aggregation [110]. Historically, it has been shown that HSP are induced in cells under stressful conditions[111]. However, they also possess an essential role under normal conditions. Their normal functions include 1. assisting the folding of newly translated proteins, 2. guiding translocating proteins through organelle membranes via action at cis and trans sides, 3. deconstructing oligomeric protein structures, 4. facilitating degradation of unstable proteins via proteolysis, 5. controlling the biological activity of folded regulatory proteins such as transcription factors[111-113]. These activities rely on ATP-regulated association of short hydrophobic segments belonging to substrate polypeptides with the HSP70 protein [114].
Substrates reacting with HSP70 to assist their folding undergo repeated cycles of binding/release [115], usually with a 1:1 stoichiometry[111]. The binding of HSP70 to its substrate acts locally rather than inducing a global conformational change [111]. According to Bukau and Horwich, all HSP70 proteins consist of the same functional parts: “a highly conserved NH2-terminal ATPase domain of 44 kDa and a COOH-terminal region of 25 kDa, divided into a conserved substrate binding domain of 15 kDa and a less-conserved immediate COOH-terminal domain of 10 kDa”[111] [figure 1].

Figure 1. Domain Organization of DnaK, DnaJ, and GrpEIndividual domains of DnaK, DnaJ, and GrpE, residue numbers defining the approximate domain borders, known structural features, and functions of domains. The definition of domains is based on 3D-structures and sequence alignments using standard algorithms. DnaK: residues 386–392 constitute a linker between the ATPase and the substrate binding domain. GrpE: residues 86–88 constitute a break of the long NH2-terminal α helix in the GrpE monomer that interacts with DnaK[111].

Through the use of biochemical and crystallographic studies, the molecular basis for HSP70 binding to its many substrates has been determined to be via the bacterial cytoplasmic homolog, DnaK [111] [Figure 2]. Studies using varying polypeptides have shown that DnaK binds most strongly to short hydrophobic segments in extended conformation[116, 117]. Thus for a substrate to be acted upon by HSP70, it must have at least a single recognizable string of residues present as an unfolded loop [111]. A consensus motif recognized by DnaK in substrates is a hydrophobic core of 4-5 residues, which is flanked by basic residues [118]. Substrate binding for HSP70 is regulated by the binding of ATP to the NH2-terminal domain of Hsp70, which causes conformational changes in the COOH-terminal binding domain[111].

Figure 2: Substrate Binding Domain of DnaK complexed with a substrate, in ribbon (A and C) and space-filling (B and D) models. A and B show a standard view, while C and D are viewed 90° counterclockwise around the vertical axis. [117].

It has been shown that members of the HSP family are important regulators of cellular survival, and may used as potential therapeutic targets for a variety of cellular injuries such as neuronal ischemia [119-121]. In recent studies it has been reported that prostaglandin E1 (PGE1) induced HSP70, HSP86, and glucose-regulated protein 78 (GRP 78) immediately after hepatic ischemia reperfusion [122]. It has recently been suggested that an associated increase of HSP70 in lithium treated rats helped to decrease infarct volume in a rat model of permanent focal cerebral ischemia[123].



Taurine: Overview and metabolism:
Taurine was first discovered in the bile of the Bos Taurus Ox, after which it was named[124]. It is an essential b-amino acid and is formally known as 2-aminoethanesulphonic acid[125]. It is prevalent in high concentrations of the cells of algae and species of the animal kingdom, but is detectable only in trace amounts or completely absent in bacteria and plants [126]. Taurine has been measured at concentrations of 0.013µmol/g wet wt in pumpkin seeds, and in a range of nuts such as almonds and walnuts at concentrations varying up to 0.046µmol/g wet wt[127]. Concentrations of taurine were undetectable in a variety of food grains and nuts, including rice, corn, wheat, barley, lentils and peanuts [128]. It is one of the most plentiful low-molecular-weight organic structures of many animals including mammals; it is estimated that a 70kg human contains 70g of taurine[126]. Taurine is ubiquitous in distribution within mammals, with tissue concentrations usually measured in the 1micromol/g wet wt range [126]. Bodily fluids such as cerebrospinal fluid, blood plasma, and extraceulluar fluids possess lower concentrations of taurine, ranging from 10-100 µM[126]. The highest concentrations are typically observed in the brain and heart, whilst the bulk of taurine mass is located in muscle tissue [126]. Neonates possess a taurine concentration in the brain 3x greater than adult brains, as concentration levels decrease as the infant develops[129]. Retinal tissue taurine concentrations exceed those found in the brain across all species studied, even in species which have low taurine levels in other tissues[130].
Molecular actions:
It has important roles in a range of physiological functions, including bile salt synthesis and conjugation with bile acids, modulation of calcium levels, maintenance of osmolarity within marine invertebrates, energy storage in marine worms, antioxidation, stabilization of membranes, and neuroinhibition in the central nervous system[124, 125]. Taurine is a sulfonic amino acid, possessing an acidic dissociation constant (pKa) similar to hydrochloric acid [126]. At physiological pH of 7.4, the high acidity causes taurine to be almost entirely zwitterionic; this gives it a high water solubility and a low degree of lipophillic attraction[126]. Due to its lipophobic nature taurine diffuses slowly across cellular membranes, hence it relies on a saturable active transport system whereby one molecule of taurine is co-transported with 1 to 3 sodium ions for transport across membranes [126, 131]. The energy cost to the cell is determined by the operation of the Na+-K+-ATPase pump, which hydrolyses one ATP molecule in order to pump out 3 sodium molecules[126]. The impermeability of bi-lipid membranes to taurine has been suggested as the mechanism accounting for the observation of large taurine concentration gradients across such membranes[126]. A concentration gradient of taurine of 1:400 has been observed in the retina[132]. This characteristic of taurine also allows it to function as a part of a hormonally-controlled active transport system operating in renal tubular fluid, which is responsible for regulation of whole body content in mammals[133-135]. Active transport processes linked to either hormonal or neurotransmitter control involving taurine have also been established in the heart and salivary glands[136], the pineal gland[137, 138], astrocytes[139], and glial cells[140]. Due to its zwitterionic characteristic, taurine has a high dipole [126]. The membrane modulatory effects of taurine and its interactions with cations such as Ca2+ have been suggested to arise from its unique ionic properities[126].
Excretion:
Taurine is considered to be biochemically inert in animals because the majority of taurine is excreted in an unaltered form[126]. It is one of the end-products in the cycle of sulphur metabolism of animals[126]. Taurine is excreted from mammals in its native form, or in bile salts in various forms including taurcholate[126]. As sulphur is metabolised in mammals, it becomes further oxidised; mammals are incapable of sulphur reduction[126]. Because of this feature of mammalian sulphur metabolism, reduced forms of sulphur present in amino acids such as methionine and cystein are essential dietary requirements[126]. Numerous animals are no longer capable of synthesising sufficient quantities of taurine to maintain whole body levels, including felines and humans; these species rely on dietary taurine intake[131].

Fig source: [126]
Metabolic pathway of sulphur-containing amino acids in mammals. The majority of these amino acids are oxidised to cystein sulfinate via the enzyme cysteine dioxygenase. In all mammals, cysteine sulfinate is rapidly transaminated to β-sulfinyl pyruvate. Some mammals are capable of enhancing quantities of taurine synthesised by diverting flow of cysteine sulfinate to be metabolised by the enzyme cysteine sulfinate decarboxylase, however humans are not.
Animals are unable to harness the free energy released during the oxidation of sulphur, because it cannot be coupled to ATP synthesis[126]. All sulphur that passes through the transaminase pathway becomes excreted as waste[126]. However, sulphur that is oxidised via the taurine pathway can become available for other uses before eventually being excreted[126].
Osmoregulation:
One of the chief non-metabolic roles of taurine is the assistance of osmoregulation in a majority of invertebrates and fish[126]. Recently, evidence has been accumulating to suggest that taurine potentiates a similar function of osmoregulation within mammals; recent research has shown taurine can be the highest concentrated organic osmolyte in both the mammalian brain and heart[126, 141, 142]. Marine invertebrates rely on organic substances to maintain osmolarity, which constitutes 60~70% of total osmotic force[126]. Mammals predominately rely on inorganic ions such as K+ and Cl- for osmoregulation, with organic substances contributing only 20% to total intracellular osmolarity[126]. The zwitterionic nature of taurine, combined with the low energy biosynthesis cost due to it being a metabolic waste product makes it a suitable candidate for being an inorganic osmoregulatory compound[126].
Transport:
Taurine entry into a cell in a hyperosmolar environment appears to be primarily achieved via active transport[143]. This active transport process relies upon Na+ ions, which are co-transported into the cell[126]. The coupled Na+ provides the energy required for the transport, while a membrane-bound Na+ -K+-ATPase maintains the concentration gradient of Na+[126]. Taurine uptake can also be stimulated by external Cl- ions[144]. The kidneys serve as the regulatory site of whole-body taurine levels in mammals[144, 145]. This is achieved via incomplete resorption of taurine by proximal tubules, and varied by numerous hormonal and second messenger signals[144, 145].
Interactions with Ca2+:
Many processes dependent on Ca2+ are capable of being regulated by taurine[126]. The rate and extent of entry of extracellular Ca2+ into cardiac tissue during the plateau of an action potential, and the following removal of cytoplasmic Ca2+ that results in relaxation can be influenced by taurine[126, 146]. Passive diffusion of Ca2+ across membranes is reduced by taurine[126]. It has been shown that taurine is positively inotropic to heart tissue in environments of sub-physiological Ca2+, and negatively inotropic to hearts in an environment containing abundant Ca2+ [147-151]. Taurine provides protection against Ca2+ cardiomyopathy[151, 152], the initiation of cardiac arrhythmias[151, 153-156], and antagonises the negative inotropic effects of calcium channel antagonists[146]. It also helps to prevent the Ca2+ paradox[157], a phenomenon where hearts placed in a Ca2+-free environment for periods longer than a few minutes undergo severe damage when reintroduced to a physiologically normal level of Ca2+[146]. The oxygen paradox, a similar situation to the Ca2+ paradox, causes enzyme leakage, Ca2+ overload, and ventricular arrhythmias; all of these damaging consequences can be prevented by presence of taurine[158]. The presence of taurine in a vesicular compartment helps to shift the equilibrium of Ca2+ influx and efflux towards a state where the vesicle is capable of storing more Ca2+ mass[159]; it has been suggested this occurs due to taurine increasing the vesicular size[126]. Taurine is also capable of increasing the initial Vmax of Ca2+ pumps[160]; conversely, a drop in tissue taurine levels can cause a decrease in initial Vmax, of ATP-dependent Ca2+ transport which has been observed in the sarcolemma[161]. The effects that taurine can exert on Ca2+ channels are varied depending on the type of channel and analogue of taurine[126]. There are two means by which taurine alters Ca2+ channel activity; secondarily to a change in the functioning of Ca2+ binding sites of acidic phospholipids in the membrane, and by altering the kinetics of channel opening and closure by effecting hydrophilic areas in close proximity to the channel[126]. Taurine is able to increase the affinity of Ca2+ binding to plasma membranes, however it reduces the membranes’ binding capacity for Ca2+; this effect can be counteracted by the presence of cations including Ca2+ itself[126].
Interactions with phospholipids:
Taurine is capable of interacting with neutral phospholipids in the cellular membrane, by binding to sites which possess low-affinity for taurine at intracellular concetrations[126]. Taurine has a negative cooperative effect on affinity of Ca2+ binding to phosphatidylserine, causing Ca2+ to bind with an even greater affinity[126]. The increased affinity for Ca2+ -phospholipid binding afforded by taurine is antagonised by Na+ and Ca2+[126]. The antagonism offered by Na+ is itself antagonised by ATP[126].
Interactions with other proteins:
Taurine has numerous effects such as neurotransmitter properties, which could be considered to be the result of interactions with other proteins[126]. These effects with proteins could be either through the direct binding of taurine to the target protein, or act secondarily via interactions with lipids to alter the local lipid environment[126]. The effects on other proteins could be either agonistic or antagonistic: for instance, binding to the site of a biologically active ligand could inhibit the target protein, while binding to a different protein could produce a direct agonistic effect[126]. However, research into this area has yet to unambiguously demonstrate that taurine does infact interact with proteins in such a manner[126]. As such, there has yet to be a convincing biochemical demonstration of taurine binding to another protein except for its transport protein[126]. There exists electrophysiological evidence which details a possible action of taurine at proteinaceous receptor sites for inhibitory amino acids, however evidence also exists to suggest that taurine does not directly interact with these sites[126]. There has yet to be convincing experimentation to show that taurine directly binds to GABA or glycine receptors[126]. The ability of taurine to stimulate Cl- flux and to hyperpolarise membranes has yet to be established as either a direct effect on the Cl- channel or an indirect effect of membrane alteration[126].
Interactions with zinc:
Zinc is another cation with which taurine interacts[126]. Deficiency of dietary zinc leads to an increase in taurine excretion[126]. Similar to taurine, zinc acts as a membrane stabiliser; together, the substances protect against membranous peroxidation damage caused by ferrous sulfate[162]. They achieve this protection by preventing water influx across the membrane which normally occurs during peroxidation[126]. Damage can also be prevented if impermeable ions replace Na+ and Cl- [163].
Functions as an antioxidant:
Despite being an essential component of life, oxygen is also toxic to living organisms due to the oxidisability of most cellular components [126]. The equation detailing the complete oxidation of oxygen to water is as follows:
O2 + 4H+ + 4e → 2H2O
This reaction occurs in 4 steps, generating the following in order; superoxide, hydroxgen peroxide dianion, hydroxyl radical, and hydroxide ion, the first three of which are more reactive than oxygen[126]. Three enzymes offer protection from oxygen-derived free radicals in the majority of tissues; these enzymes are superoxide dismutase (converts superoxide to hydrogen peroxide), catalase, and glutathione peroxidase (both of which break down hydrogen peroxide)[126]. There exists no enzyme to assist the removal of hydroxyl radicals or hypochlorite[126]. Polyunsaturated fatty acids are one of the more oxidation-susceptible components of cellular membranes[164]. It has been theorised that the high redox potential of partially oxidised sulfur-containing amino acid intermediates, such as cysteine, cysteamine, hypotaurine and taurine could protect against oxidative damage[126]. Experimentation has shown that hypotaurine and cysteamine instantaneously oxidise, and are excellent scavengers for hydroxyl radical and hypochlorite[126, 165, 166].
Aim:
To analyse and quantify the expression of CHOP after a 4-week atherogenic diet in the endothelium of left main coronary artery of taurine-fed and statin-fed rabbits.
Hypothesis:
- Taurine-fed rabbits will feature reduced levels of CHOP due to anti-oxidative effects leading to reduced ER stress
-Statin-fed rabbits will not feature reduced levels of CHOP
Materials:
-Goat Serum purchased from CHEMICON, located in Upstate Linco. Catalogue # S26-100mL.
-CHOP antibody at a 1mg/ml concentration, purchased from Affinity BioReagents, located in Golden, USA. Catalogue # MA1-250.
-MCIDTM Core Analysis version 7.0 was purchased from “Imaging Research”, 1994-2003 all rights reserved.
-Lecia Application Suite version 2.8.1©was purchased from Lecia Microsystems Ltd.(located in Switzerland) 2003-2007
-DAB+ chromogen, DAB+ substrate buffer, and EnVision+R system labeled Polymer-HRP Anti-mouse were purchased from Dako Australia, HQ located in Denmark.

Methods:
Animal preparation:
Fifteen male New Zealand White Rabbits, 3 months of age, were randomly sorted into 3 groups. Each group was fed a different diet;
Group A (Control, Con) was fed a normal chow diet
Group B (Methionine, MC) was fed a chow diet supplemented with 0.5% cholesterol, 1% methionine and 5% peanut oil
Group C (Taurine, MCT) was fed the same diet as Group B, with an addition of 2% taurine
The rabbits were divided into two batches because of limitations on how many rabbits could be housed at one time. The division consisted of one group of 9 rabbits and then a second group of 6 rabbits. The first 9 rabbits were divided into 3 groups of 3, each group receiving diet A B or C. The second group of 6 rabbits was split into groups of 3 groups of 2, each group receiving diet A B or C.
10ml of blood was collected from each rabbit at weekly intervals, commencing at week 0 and continuing until week 4. These 10ml samples were subdivided into 2ml samples, which were then placed into a centrifuge to collect plasma. Following plasma collection, the samples were immediately frozen.

Specimen collection:
When week 4 had ended, each rabbit was weighed and the measurement was recorded. In order to prepare the rabbits for anesthesia, the skin overlying the vein on the dorsal aspect of the rabbit ear was carefully shaved of fur. An anesthetic spray and/or cream were applied to locally numb the ear. After 5 minutes, a lethal anesthetic mixture of Ilium Xylazil (20mg/ml) and Ketamine (100mg/ml) in a ratio of 3:2 was injected into the vein of the rabbit ear. The catheter used to administer the mixture was taped to the ear incase additional anesthetic was required. The following table was used to determine dosage quantity.
Table: Anesthetic Dosage
Rabbit Weight (kg) Anesthetic mixture (ml)
2 1
2.5 1.25
3 1.5
3.5 1.75
4 2
4.5 2.25
5 2.7

To test that the anesthetic had successfully retired the rabbit, the rabbit’s forepaw was squeezed and its jaw gently moved side to side. If the jaw was relaxed and easy to move, and no signs of ankle reflex were observed upon squeezing the forepaw, the rabbit was considered relieved.
Once relieved, the rabbits were dissected by cutting through the abdominal musculature in a coronal direction, ascending towards the thorax. Once the thoracic cavity was opened, several drops of heparin were applied to the surfaces of organs and tissues present within the cavity. The ascending aorta was cut, and a 10ml sample of blood was quickly collected from the aorta and placed into 5 heparin-containing tubes in 2ml quantities. Plasma was then collected from these blood samples as previously described.
The rabbit heart was detached and horizontally sectioned. The basal section (which contained the left main coronary artery) and apical sections of heart were placed into separate containers containing paraffin. The liver and kidneys were removed from the rabbit and then swiftly cut into smaller cuboidal pieces of approximately 5mm dimensions. These small pieces of tissue were then immediately placed into 4% paraformaldehyde solution for tissue fixation. Segments of the renal artery and ascending aorta were also removed from the rabbit and quickly placed into 4% paraformaldehyde solution. The entire abdominal aorta was removed from the rabbit, and carefully trimmed of surrounding connective tissue in a petri dish containing phosphate-buffered saline (PBS) and Ca2+ ions at a temperature of 37°C.
Tissue samples preserved in paraformaldehyde solution were treated with fresh paraformaldehyde the following day, and then PBS solution on the third day. After treatment with PBS solution on the third day, the tissue samples were stored in a cold room of temperature 0-4°C for later processing.

Tissue Processing:
The paraffin blocks housing the LMCA were prepared at the Deparment of Anatomy and Cell Biology, Monash University, Clayton. The tissue from taurine rabbits 1-9 were placed into one paraffin block, while the tissue from rabbits 10-14 + 18 were placed into a separate block. The tissue from statin rabbits were all placed into one paraffin block.
Immunohistochemistry:
Using a microtome, the paraffin blocks containing embedded tissue samples were sliced into 5µm sheets. They were then placed to float on the surface of a bath of water at 45°C, and collected using glass slides coated in gelatin. The slides were then left to dry prior to immunohistochemistry staining.
Slides from adjacent sections of tissue were chosen for immunohistochemistry. Care was taken to ensure that at no stage during the staining process were the slides allowed to dry. During each transference step between trays of liquid, the slide tray was quickly rinsed of excess fluid by gentle tapping.
**The following occured inside a fume-hood**
-Slides were placed securely into a slide tray.
-The slide tray was placed into a container of 100% xylene and left for 5 minutes.
-The tray was then placed into a 2nd container of 100% xylene and left for 5 minutes.
-Then the tray was placed into a container of 10% ethanol and left for 1 minute.
-The tray was then placed into a 2nd container of 100% ethanol and left for 1 minute.
-The tray was then placed into a 3rd container of ethanol, this time at a concentration of 90%. They were left for 1 minute.
-The tray was then placed into a container of H2O for a brief wash.
-The slide tray was then removed from the fume-hood and placed into a container of 10mM Tris buffer for 5 minutes.
-During these 5 minutes, the blocking solution was prepared. The blocking solution was comprised of a 1:100 dilution of frozen goat serum in 10mM Tris buffer. To make the blocking solution, 5ml of Tris buffer was added to a vial containing 50µL of frozen goat serum. The vial of blocking solution was shaken to ensure all of the goat serum had thawed and dissolved.
-The slides were removed from the Tris buffer and quickly drained of excess fluid. A wax pen was used to demarcate the borders around the artery specimens where the blocking solution was to be applied. Approximately 400µL of blocking solution was added to each slide, and gently swirled around to ensure homogenous spread.
-The slides were then placed into a humidity chamber to stand.
-Water was added to the humidity chamber. Care was taken to ensure the water level didn’t rise to reach the slides. A lid was applied to the chamber to ensure a humidified environment for the slides. The slides were left in the chamber for 20 minutes.
-During these 20 minutes, the primary anti-body was prepared. CHOP was used. The CHOP was diluted at a 1:1000 ratio in the blocking solution. Approximately 500µL of diluted CHOP was prepared for each slide.
-After 20 minutes, the slides were removed from the humidity chamber and quickly drained of blocking solution. The CHOP antibody was then applied to each slide, with gentle swirling to ensure a homogenous spread. The slides were placed back into the humidity chamber and left overnight at room temperature.
-The following day, the slides were removed from the humidity chamber and rinsed in a container of 10mM Tris for 5 minutes.
-After 5 minutes, the slides were gently rinsed. Approximately 400µL of undiluted EnVision anti-mouse antibody was then added to each slide. The slides were placed back into the humidity chamber and left to stand for 1 hour.
-The slides were then placed into a container of 10mM Tris to rinse for 5 minutes.
-During the 5 minute rinse, a mixture of DAB and substrate buffer was prepared. Approximately 1 droplet of DAB was added to 1.5ml of substrate buffer and mixed in an eppendorf tube.
-After 5 minutes, the slides were gently rinsed. Approximately 400µL of diluted DAB mixture was applied to the slides, gently spread, and left for 1 minute. A brown colour developed.
-After 1 minute, the slides were briefly washed in a container of H2O for 1 minute.
-The slides were then rinsed of H2O, and filtered haematoxylin was added to the slides. Enough haematoxylin was added to adequately cover the tissue on the slides. The slides were left to stand for 1 minute, and then washed in H2O for an additional minute.
**The following steps occur inside a fume-hood**
-Following the wash, the slides were placed into a container of Scotts tap-water for approximately 1 minute. A blue colour developed.
-The slides were then washed in H2O for 1 minute.
-The slides were then placed into a container of 90% ethanol for 1 minute.
-The slides were then transferred to a container of 100% ethanol for 1 minute.
- After 1 minute, the slides were placed into a 2nd container of 100% ethanol for 1 minute.
-The slides were then washed in a container of xylene for 1 minute to remove the ethanol.
-The slides were then placed to stand in a container of xylene for 5 minutes.
-After 5 minutes, the slides were placed into a 2nd container of xylene and left for an additional 5 minutes.
-During these 5 minutes, cover-slips for the slides were prepared by applying droplets of DPX mounting media. These cover-slips were placed on a cloth with the DPX droplets exposed.
-The slides were removed from the xylene one by one and placed face-down onto the DPX-covered cover slips. Firm pressure was applied to the slide to ensure no air-bubbles were fixed within the DPX. The slides were left overnight to dry inside the fume-hood.
Image capture:
Slides were placed under a light microscope. The microscope was linked to a computer featuring Lecia application suite, a program capable of capturing images from the light microscope. The focus on the microscope was set to 40x. The program settings of light exposure, saturation, gamma, and gain were maintained for all photos captured from one particular slide. White balance function was used prior to the first image capture to ensure high-quality images were photographed. Four images were captured of each artery, which were chosen randomly and arbitrarily named “top”, “bottom”, “left”, and “right”. In some cases, a segment of artery wall was absent; in these instances, only three images were captured. All images were appropriately named and saved to hard drive as .tif files. They were then transferred to another computer for analysis with MCID software.

Image Analysis:
Images captured were retrieved in the MCID program. The program enabled the tracing of endothelium in each image, and was capable of detecting color of a certain light intensity, saturation, and hue which was calibrated manually prior to tracing. The program was calibrated to detect areas of intensely stained brown-colored DAB. Each image was traced at 100% magnification, and the parameters of “color intensity” and “proportional area” were recorded into Microsoft Excel. The tracing was executed using a “click-ribbon” tool, which appeared on-screen as a circle. By left-clicking at one point, then dragging the mouse to another location and left-clicking again, a “ribbon” was created between the two points. The diameter of the ribbon was altered by holding the Ctrl key; care was taken to ensure the points selected and ribbon thickness matched the endothelium as accurately as possible. Once the endothelium of an image had been traced, double-clicking the left mouse button allowed the program to scan the area inside of the ribbon for colors fitting the pre-set critera. Only areas of in-tact endothelium were traced: spots where denudation had occurred were “ignored” by creating a pixel-thin ribbon to connect adjacent segments of endothelium. The same color detection parameters were maintained across a set of tracings for all of the arteries from a particular slide. Because the statin arteries were all on one slide, the same parameters were kept for all tracings of these images. For the taurine group of arteries two sets of parameters were used: One for images obtained from arteries 1-9, and another for arteries obtained from arteries 10-14 + 18. After all the images had been traced, the parameters were re-calibrated and another tracing of the images was then repeated. Repeat tracings for all images were conducted until significant results were obtained: For the taurine group, this required two tracings. For the statin group, three tracings were needed.
Data Analysis:
All data collected from image tracings was entered into Microsoft Excel. The following format for entering data was used:
Image name: Intensity (I): Proportionate area (PA): 1-I: (1-I) *PA: …*100 (Xn) Average of tracings for artery:
Artery 7 left 0.3579 0.0131 0.6421 0.00841151 0.841151 0.415266
Artery 7 right 0.3764 0.0011 0.6236 0.00068596 0.068596
Artery 7 top 0.3744 0.0023 0.6256 0.00143888 0.143888
Artery 7 bottom 0.3538 0.0094 0.6462 0.00607428 0.607428

The average was calculated as follows: (XLeft + XRight + XTop + XBottom)/4.
For arteries which lacked four separate images, data-fields for “missing” images were filled as “BLANK”. The average value calculated for these arteries used only X values which were present.
Image name: Intensity (I): Proportionate area (PA): 1-I: (1-I) *PA: …*100 (Xn) Average of tracings for artery:
Artery 3 left BLANK BLANK 1.658264
Artery 3 right 0.3605 0.0163 0.6395 0.01042385 1.042385
Artery 3 top 0.3581 0.0369 0.6419 0.02368611 2.368611
Artery 3 bottom 0.3591 0.0244 0.6409 0.01563796 1.563796

The average values obtained for each set of images was then tabulated and normalised against the control group averages. Because the taurine rabbit group images came from two separate slides featuring two different sets of MCID tracing parameters, two normalisation processes were required. The following table describes how this was performed:
MCT averages: MC averages: CON averages:
Artery 1 Artery 4 Artery 7
Artery 2 Artery 5 Artery 8
Artery 3 Artery 6 Artery 9
Artery 10 Artery 12 Artery 14
Artery 11 Artery 13 Artery 18

The values of artery 7 8 and 9 were averaged. The value obtained was used to normalise arteries 1-9.
The values of artery 14 and 18 were averaged. The value obtained was used to normalise arteries 10-14 + 18.
The normalised data was then analysed using ANOVA analysis and 2-tailed Student unpaired t test. Means and standard errors were also calculated for each set of normalised data. Results were graphed as mean ± SED. A probability value <0.05 was considered significant.
1. Organisation, W.H., Cardiovascular diseases fact sheet 2007. February 2007, World Health Organisation.
2. Tegos, T.J.M., The Genesis of Atherosclerosis and Risk Factors: A Review. Angiology, Volume 52(2), February 2001: p. 89-98.
3. Xu, Q., Mechanisms of vascular disease. 2007, Cambridge: Cambridge University Press.
4. More RH, M.H., Haust MD, Role of mural fibrin thrombi of the aorta in genesis of arteriosclerotic plaques. A.M.A. archives of pathology, 1957. 63: p. 612-20.
5. AM, M., Atherosclerosis. Pediatrics, 1987. 79: p. 651-653.
6. Napoli C, D.A.F., Mancini FP, et al, Fatty streak formation occurs in human fetal aortas and is greatly enhanced by maternal hypercholesterolemia: intimal accumulation of low density lipoprotein and its oxidation precede monocyte recruitment into early atherosclerotic lesions. The Journal of Clinical Investigation, 1997. 100: p. 2680-2690.
7. Stary HC, C.A., Glagov S, et al, A definition of initial, fatty streak, and intermediate lesions of atherosclerosis: a report from the Committee on Vascular Lesions of the Council on Arteriosclerosis. Circulation, 1994. 85(5): p. 2462-2479.
8. Stary, H.C., Lipid and macrophage accumulations in arteries of children and the development of atherosclerosis. The American Journal of Clinical Nutrution, November, 2000: p. 1297S-1306S.
9. Thomas J. Tegos, M., Evi Kalodiki, PhD, Michael M. Sabetai, MD, Andrew N. Nicolaides, MS, The genesis of Atherosclerosis and Risk Factors: A Review. Angiology, 2001. 52(2): p. 89-98.
10. Campbell GR, C.J., Smooth muscle phenotypic changes in arterial wall homeostasis: Implications for the pathogenesis of atherosclerosis. Exp Mol Pathol, 1985. 42: p. 139-162.
11. Bostrom K, W.K., Stanford WP, et al, Atherosclerotic calcification: Relation to developmental osteogenesis. American Journal of Cardiology, 1995. 75: p. 88B-91B.
12. H.C. Stary, A.B.C., R.E. Dinsmore et al, A definition of advanced types of atherosclerotic lesions and a histological classification of atherosclerosis. A report from the Committee on Vascular Lesions of the Council on Arteriosclerosis, American Heart Association. Arteriosclerosis, Thrombosis and Vascular Biology, 1995. 15: p. 1512-31.
13. Stary, H.C., Lipid and macrophage accumulations in arteries of children and
the development of atherosclerosis1–. The American Journal of Clinical Nutrition, 2000: p. 1297S - 1306S.
14. P. Libby, P.M.R.A.M., Inflamation and atherosclerosis. Circulation, 2002: p. 1135-1143.
15. H.C. Stary, A.B.C., R.E. Dinsmore et al, A definition of advanced types of atherosclerotic lesions and histological classification of atherosclerosis. A report from the Committee on Vascular Lesions of the Council on Arteriosclerosis, American Heart Association. Arteriosclerosis, Thrombosis, and Vascular Biology, 1995: p. 1512 - 1531.
16. Ross, R., Atherosclerosis - An Inflammatory Disease. New England Journal of Medicine, 1999. 340: p. 115-126.
17. Goldman, e.a., Cecil's Textbook of Medicine. 22nd ed. 2004.
18. Witztum, C.K.G.J.L., Atherosclerosis: the road ahead. Cell, 2001. 104: p. 503-516.
19. D, S., Low density lipoprotein oxidation and its pathobiological significance. The American Society for Biochemistry and Molecular Biology, Inc., 1997. 272: p. 20963-20966.
20. Navab M, B.J., Watson AD, et al, The Yin and Yang of oxidation in the development of the fatty streak: a review based on the 1994 George Lyman Duff Memorial Lecture. Arteriosclerosis, Thrombosis and Vascular Biology, 1996. 16: p. 831-842.
21. Nunes GL, R.K., Kalynych A, King SB III, Sgoutas DS, Berk BC, Vitamins
C and E inhibit O2-production in the pig coronary artery. Circulation, 1997. 96: p. 3593-3601.
22. Quinn MT, P.S., Fong LG, Steinberg D, Oxidatively modified low density lipoproteins: a potential role in recruitment and retention of monocyte/macrophages during atherogenesis. Proceedings of the National Academy of Sciences of the United States of America, 1987. 84: p. 2995-2998.
23. Leonard EJ, Y.T., Human monocyte chemoattractant protein-1 (MCP-1). Immunology Today, 1990. 11: p. 97-101.
24. Stopeck AT, N.A., Mancini FP, Hajjar DP, Cytokine regulation of low density lipoprotein receptor gene transcription in HepG2 cells. Journal of Biological Chemistry, 1993. 268: p. 17489-17494.
25. Hajjar DP, H.M., Lipoprotein trafficking in vascular cells:
molecular Trojan horses and cellular saboteurs. Journal of Biological Chemistry, 1997. 272: p. 22975-22978.
26. Falcone DJ, M.T., Vergilio JA, Stimulation of macrophage urokinase expression by polyanions is protein kinase C-dependent and requires protein and RNA synthesis. Journal of Biological Chemistry, 1991. 266: p. 22726-22732.
27. Geng Y-J, L.P., Evidence for apoptosis in advanced human atheroma: colocalization with interleukin-1 beta-converting enzyme. American Journal of Pathology, 1995. 147: p. 251-266.
28. T, P., Induction of interleukin-1 production by ligands binding to the scavenger receptor in human monocytes and the THP-1 cell line. Immunology, 1991. 74(3): p. 432-438.
29. Palkama T, M.S., Hurme M, Tyrosine kinase activity is involved in the protein kinase C induced expression of interleukin-1 beta gene in monocytic cells. FEBS Letters, 1993. 319: p. 100-104.
30. Shepherd J, C.S., Ford I, et al, Prevention of coronary heart disease with pravastatin in men with hypercholesterolemia. New England Journal of Medicine, 1995. 333: p. 1301-1307.
31. JL, B., Cardiovascular disease burden increases, NIH funding decreases. Nat Med, 1997. 3: p. 600-601.
32. Lecture, B.E.S., Cardiovascular medicine at the turn of the millennium: triumphs, concerns, and opportunities. New England Journal of Medicine, 1997. 337: p. 1360-1369.
33. MR, M., Plasma homocyst(e)ine and arterial occlusive diseases: a mini-review. Clinical Chemistry, 1995. 41: p. 173-176.
34. KS, M., Vascular pathology of homocysteinemia: implications for the pathogenesis of arteriosclerosis. American Journal of Pathology, 1969. 56: p. 111-128.
35. Mudd SH, S.F., Levy HL, et al, The natural history of homocystinuria due to cystathionine (beta)-synthase deficiency. American Journal of Human Genetics, 1985. 37: p. 1-31.
36. Nehler MR, T.L.J., Porter JM, Homocysteinemia as a risk factor for atherosclerosis: a review. Cardiovascular Surgery, 1997. 6: p. 559-567.
37. Nygard O, N.J., Refsum H, Ueland PM, Farstad M, Vollset SE, Plasma homocysteine levels and mortality in patients with coronary artery disease. New England Journal of Medicine, 1997. 337: p. 230-236.
38. Harker LA, R.R., Slichter SJ, Scott CR., Homocystine-induced arteriosclerosis: the role of endothelial cell injury and platelet response in its genesis. The Journal of Clinical Investigation, 1976. 58: p. 731-741.
39. KA, H., Homocysteine-induced modulation of tissue plasminogen activator binding to its endothelial cell membrane receptor. The Journal of Clinical Investigation, 1993. 91: p. 2873-2879.
40. Majors A, E.L., Pezacka EH, EH. Homocysteine as a risk factor for vascular disease: enhanced collagen production and accumulation by smooth muscle cells. Arteriosclerosis, Thrombosis and Vascular Biology, 1997. 17: p. 2074-2081.
41. Upchurch GR Jr, W.G., Fabian AJ, et al, Homocyst(e)ine decreases bioavailable nitric oxide by a mechanism involving glutathione peroxidase. Journal of Biological Chemistry, 1997. 272: p. 17012-17017.
42. Omenn GS, B.S., Motulsky AG, Preventing coronary heart disease: B vitamins and homocysteine. Circulation, 1998. 97: p. 421-444.
43. Barbara Glowinska, M.U., Alicja Koput, Marzena Galar, New atherosclerosis risk factorsnext term in previous termobese, hypertensive and diabetic children and adolescents. Atherosclerosis, 2003. 167(2): p. 275-286.
44. Yla-Herttuala S, P.W., Rosenfeld ME, et al, Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man. The Journal of Clinical Investigation, 1989. 84: p. 1086-1095.
45. Navab M, B.J., Watson AD, et al, The Yin and Yang of oxidation in the development of the fatty streak: a review based on the 1994 George Lyman Duff Memorial Lecture. Arteriosclerosis, Thrombosis and Vascular Biology, 1996. 16: p. 831-842.
46. Chang MY, S.M., Chait A, Raines EW, Ross R, Inhibition of hypercholesterolemia-induced atherosclerosis in the nonhuman primate by probucol. II. Cellular composition and proliferation. Arteriosclerosis, Thrombosis and Vascular Biology, 1995. 15: p. 1631-1640.
47. Fruebis J, G.V., Silvestre M, Palinski W. , Effect of probucol treatment on gene expression of VCAM-1, MCP-1, and M-CSF in the aortic wall of LDL receptor-deficient rabbits during early atherogenesis. Arteriosclerosis, Thrombosis and Vascular Biology, 1997. 17: p. 1289-1302.
48. Reaven PD, K.A., Beltz WF, Parthasarathy S, Witztum JL., Effect of dietary antioxidant combinations in humans: protection of LDL by vitamin E but not by beta-carotene. Arteriosclerosis, Thrombosis and Vascular Biology, 1993. 13: p. 590-600.
49. Hennekens CH, B.J., Manson JE, et al, Lack of effect of long-term supplementation with beta carotene on the incidence of malignant neoplasms and cardiovascular disease. New England Journal of Medicine, 1996. 334: p. 1145-1149.
50. Ross R, G.J., Atherosclerosis and the arterial smooth muscle cell: proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science, 1973. 180: p. 1332-1339.
51. Glagov S, W.E., Zarins CK, Stankunavicius R, Kolettis GJ, Compensatory enlargement of human atherosclerotic coronary arteries. New England Journal of Medicine, 1987. 316: p. 1371-1375.
52. Fuster V, R.R., Topol EJ, Atherosclerosis and coronary artery disease. Cytokines and growth regulatory molecules, ed. R.R. Libby P. Vol. 1. 1996, Philadelphia: Lippincott-Raven.
53. Fuster V, R.R., Topol EJ, Atherosclerosis and coronary artery disease. Pathogenesis of plaque disruption, ed. S.P. Falk E, Fuster V. Vol. 2. 1996: Lippincott-Raven.
54. GK, H., Inflammation, atherosclerosis, and coronary artery disease. New England Journal of Medicine, 2005. 352: p. 1685-1695.
55. LH, S., Atherosclerosis: an immunologically mediated (autoimmune?) disease. Journal of Clinical Rheumatology, 2007. 13: p. 160-168.
56. Proh´aszka Z, F.u.G., Immunological aspects of heat-shock proteins—the optimum stress of life. Molecular Immunology, 2004. 41: p. 29-44.
57. Xiao Q, M.K., Schett G, et al, Association of serum-soluble heat shock protein 60 with carotid atherosclerosis. Stroke, 2005. 36: p. 2571-2576.
58. Mandal K, J.M., Xu Q, The immune response in atherosclerosis: a double edged sword. Nature reviews Immunology, 2006. 6: p. 508-518.
59. Sherer Y, S.Y., Mechanisms of disease: atherosclerosis in autoimmune diseases. Nature Clinical Practice Rheumatology, 2006. 2: p. 99-106.
60. Jonathan H. Lin, P.W., T.S. Bendict Yen, Endoplasmic Reticulum Stress in Disease Pathogenesis. Annual Review of Pathology: Mechanisms of Disease, 2008. 3: p. 399-425.
61. RJ., K., Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes & Development, 1999. 13(10): p. 1211-1233.
62. Lee AS, D.A., Baker V, Chow PC, Transcriptional regulation of two genes specifically induced by glucose starvation in a hamster mutant fibroblast cell line. Journal of Biological Chemistry, 1983. 258: p. 597-603.
63. Lee AS, D.A., Baker V, Chow PC, Coordinated regulation of a set of genes by glucose and calcium ionophore in mammalian cells. Trends in Biochemical Sciences 1987. 12: p. 20-24.
64. Haas IG, W.M., Immunoglobulin heavy chain binding protein. Nature, 1983. 306: p. 387-389.
65. Gething MJ, S.J., Protein folding in the cell. Nature, 1992. 355: p. 33-45.
66. Kezhong Zhang, R.J.K., The unfolded protein response: A stress signaling pathway critical for health and disease. Neurology, 2006. 66(2 Supplement 1): p. S102-109.
67. Cox JS, S.C., Walter P., Transcriptional induction of genes encoding endoplasmic reticulum resident proteins requires a transmembrane protein kinase. Cell, 1993. 73: p. 1197-1206.
68. Mori K, M.W., Gething M-J, Sambrook J., A transmembrane protein with a cdc2+/CDC28-related kinase activity is required for signalling from the ER to the nucleus. Cell, 1993. 74: p. 743-756.
69. Sidrauski C, C.J., Walter P, tRNA ligase is required for regulated mRNA splicing in the unfolded protein response. Cell, 1996. 87: p. 405-413.
70. Sidrauski C, W.P., The transmembrane kinase Ire1p is a sitespecific endonuclease that initiates mRNA splicing in the unfolded protein response. Cell, 1997. 90: p. 1031-1039.
71. Kawahara T, Y.H., Yura T, Mori K., Unconventional splicing of HAC1/ERN4 mRNA required for the unfolded protein response. Sequence-specific and non-sequential cleavage of the splice sites. Journal of Biological Chemistry, 1998. 273: p. 1802-1807.
72. Travers KJ, P.C., Wodicka L, Lockhart DJ, Weissman JS, Walter P, Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell, 2000. 101: p. 249-258.
73. Tirasophon W, W.A., Kaufman RJ., A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells. Genes & Development, 1998. 12: p. 1812-1824.
74. Wang XZ, H.H., Zhang Y, Jolicoeur EM, Kuroda M, Ron D., Cloning of mammalian Ire1 reveals diversity in the ER stress responses. The EMBO Journal, 1998. 17: p. 5708-5717.
75. Shi Y, V.K., Sood R, An J, Liang J, Stramm L, et al, Identification and characterization of pancreatic eukaryotic initiation factor 2 alpha-subunit kinase, PEK, involved in translational control. Molecular and Cellular Biology, 1998. 18: p. 7499-7509.
76. Haze K, Y.H., Yanagi H, Yura T, Mori K., Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Molecular and Cellular Biology, 1999. 10: p. 3787-3799.
77. Bertolotti A, Z.Y., Hendershot LM, Harding HP, Ron D., Dynamic interaction of BiP and ER stress transducers in the unfolded- protein response. Nature Cell Biology, 2000. 2: p. 326-332.
78. Liu CY, X.Z., Kaufman RJ, Structure and intermolecular interactions of the luminal dimerization domain of human IRE1alpha. Journal of Biological Chemistry, 2003. 278: p. 17680-17687.
79. Shen J, C.X., Hendershot L, Prywes R., ER stress regulation of ATF6 localization by dissociation of BiP/GRP78 binding and unmasking of Golgi localization signals. Developmental Cell, 2002. 3: p. 99-111.
80. Zhang K, W.H., Song B, Miller CN, Scheuner D, Kaufman RJ., The unfolded protein response sensor IRE1alpha is required at 2 distinct steps in B cell lymphopoiesis. The Journal of Clinical Investigation, 2005. 115: p. 268-281.
81. Yoshida H, H.K., Yanagi H, Yura T, Mori K, Identification of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose- regulated proteins. Involvement of basic leucine zipper transcription factors. Journal of Biological Chemistry, 1998. 273: p. 33741-33749.
82. Yoshida H, M.T., Yamamoto A, Okada T, Mori K, XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell, 2001. 107: p. 881-891.
83. Shen X, E.R., Lee K, Liu CY, Yang K, Solomon A, et al., Complementary signaling pathways regulate the unfolded protein response and are required for C. elegans development. Cell, 2001. 107: p. 893-903.
84. Calfon M, Z.H., Urano F, Till JH, Hubbard SR, Harding HP, et al., IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature, 2002. 415: p. 92-96.
85. Yoshida H, O.T., Haze K, et al., ATF6 Activated by Proteolysis Binds in the Presence of NF-Y (CBF) Directly to the cis-Acting Element Responsible for the Mammalian Unfolded Protein Response. Molecular and Cellular Biology, 2000. 20: p. 6755-6767.
86. Li M, B.P., Roy B, et al., ATF6 as a transcription activator of the endoplasmic reticulum stress element: thapsigargin stress-induced changes and synergistic interactions with NF-Y and YY1. Molecular and Cellular Biology, 2000. 20: p. 5096-5106.
87. Ye J, R.R., Komuro R, et al., ER Stress Induces Cleavage of Membrane-Bound ATF6 by the Same Proteases that Process SREBPs. Molecular Cell, 2000. 6: p. 1355-1364.
88. Okada T, Y.H., Akazawa R, Negishi M, Mori K., Distinct Roles of ATF6 and PERK in Transcription during the Mammalian Unfolded Protein Response. Biochemical Journal, 2002. 366: p. 585-594.
89. Lee K, T.W., Shen X, et al, IRE1-mediated unconventional mRNA splicing and S2P-mediated ATF6 cleavage merge to regulate XBP1 in signaling the unfolded protein response. Genes & Development, 2002. 16: p. 452-466.
90. Lee AH, I.N., Glimcher LH, XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response. Molecular and Cellular Biology, 2003. 23: p. 7448-7459.
91. Harding HP, Z.H., Zhang Y, Jungries R, Chung P, Plesken H, et al., Diabetes mellitus and exocrine pancreatic dysfunction in perk-/- mice reveals a role for translational control in secretory cell survival. Molecular Cell, 2001. 7: p. 1153-1163.
92. Harding HP, Z.Y., Bertolotti A, Zeng H, Ron D., Perk is essential for translational regulation and cell survival during the unfolded protein response. Molecular Cell, 2000. 5: p. 897-904.
93. RJ, K., Regulation of mRNA translation by protein folding in the endoplasmic reticulum. Trends in Biochemical Sciences, 2004. 29: p. 152-158.
94. Scheuner D, S.B., McEwen E, et al, Translational control is required for the unfolded protein response and in vivo glucose homeostasis. Molecular Cell, 2001. 7: p. 1165-1176.
95. Harding HP, Z.Y., Bertolotti A, Zeng H, Ron D., Perk is essential for translational regulation and cell survival during the unfolded protein response. Molecular Cell, 2000. 5: p. 897-904.
96. Harding HP, Z.Y., Zeng H, Novoa I, Lu PD, Calfon M, et al., An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Molecular Cell, 2003. 11: p. 619-633.
97. Dorner AJ, W.L., Kaufman RJ., Protein dissociation from GRP78 and secretion are blocked by depletion of cellular ATP levels. Proceedings of the National Academy of Sciences of the United States of America, 1990. 87: p. 7429-7432.
98. Werner ED, B.J., McCracken AA, Proteasome-dependent endoplasmic reticulum-associated protein degradation: an unconventional route to a familiar fate. Proceedings of the National Academy of Sciences of the United States of America, 1996. 93: p. 13797-13801.
99. Hosokawa N, W.I., Hasegawa K, et al, A novel ER alphamannosidase- like protein accelerates ER-associated degradation. EMBO reports, 2001. 2: p. 415-422.
100. Molinari M, C.V., Galli C, Lucca P, Paganetti P, Role of EDEM in the release of misfolded glycoproteins from the calnexin cycle. Science, 2003. 299: p. 1397-1400.
101. Oda Y, H.N., Wada I, Nagata K., EDEM as an acceptor of terminally misfolded glycoproteins released from calnexin. Science, 2003. 299: p. 1394-1397.
102. Yoshida H, M.T., Hosokawa N, Kaufman RJ, Nagata K, Mori K., A time-dependent phase shift in the mammalian unfolded protein response. Developmental Cell, 2003. 4: p. 265-271.
103. Urano F, W.X., Bertolotti A, et al., Coupling of Stress in the ER to Activation of JNK Protein Kinases by Transmembrane Protein Kinase IRE1. Science, 2000. 287: p. 664-666.
104. Yoneda T, I.K., Oono K, et al, Activation of Caspase-12, an Endoplastic Reticulum (ER) Resident Caspase, through Tumor Necrosis Factor Receptor-associated Factor 2- dependent Mechanism in Response to the ER Stress. Journal of Biological Chemistry, 2001. 276: p. 13935-13940.
105. Nishitoh H, M.A., Tobiume K, et al, ASK1 is essential for endoplasmic reticulum stress-induced neuronal cell death triggered by expanded polyglutamine repeats. Genes & Development, 2002. 16: p. 1345-1355.
106. Leppa S, B.D., Diverse functions of JNK signaling and c-Jun in stress response and apoptosis. Oncogene, 1999. 18: p. 6158-6162.
107. Nakagawa T, Z.H., Morishima N, et al, Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloidbeta. Nature, 2000. 403: p. 98-103.
108. Morishima N, N.K., Takenouchi H, Shibata T, Yasuhiko Y., An ER stress-specific caspase cascade in apoptosis: cytochrome c- independent activation of caspase-9 by caspase-12. Journal of Biological Chemistry, 2002. 277: p. 34287-34294.
109. Rong HAN, B.G., Rui SHENG, Li-sha ZHANG, Hui-lin ZHANG, Zhen-lun GU, Zheng-hong QIN, Synergistic effects of prostaglandin E1 and lithium in a rat model of cerebral ischemia,. Acta Pharmacologica Sinica 2008. 29(10): p. 1141-1149.
110. Ohtsuka1 K, S.T., Roles of molecular chaperones in the nervous system. Brain Research Bulletin, 2000. 53(2): p. 141-146.
111. Bernd Bukau, A.L.H., Hsp70 and Hsp60 Chaperone Machines. Cell, 1998. 92(3): p. 351-366.
112. R.I. Morimoto, A.T.a.C.G., The Biology of Heat Shock Proteins and Molecular Chaperones. 1994, Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press.
113. Hartl, F.U., Molecular chaperones in cellular protein folding. Nature 1996. 381: p. 571-580.
114. G.C. Flynn, J.P., M.T. Flocco and J.E. Rothman, Peptide-binding specificity of the molecular chaperone BiP. Nature, 1991. 353: p. 726-730.
115. A. Szabo, T.L., H. Schröder, J. Flanagan, B. Bukau and F.U. Hartl, The ATP hydrolysis-dependent reaction cycle of the Escherichia coli Hsp70 system-DnaK, DnaJ and GrpE. Proceedings of the National Academy of Sciences of the United States of America, 1994. 91(22): p. 10345-10349.
116. D. Schmid, A.B., H. Gehring and P. Christen, Kinetics of molecular chaperone action. Science, 1994. 263: p. 971-973.
117. X. Zhu, X.Z., W.F. Burkholder, A. Gragerov, C.M. Ogata, M. Gottesman and W.A. Hendrickson, Structural analysis of substrate binding by the molecular chaperone DnaK. Science, 1996. 272: p. 1606-1614.
118. S. Rüdiger, L.G., J. Schneider-Mergener and B. Bukau, Substrate specificity of the DnaK chaperone determined by screening cellulose-bound peptide libraries. EMBO Journal, 1997. 16: p. 1501-1507.
119. Tsuchiya D, H.S., Matsumori Y, Shiina H, Kayama T, Swanson RA, et al. , Overexpression of rat heat shock protein 70 is associated with reduction of early mitochondrial cytochrome C release and subsequent DNA fragmentation after permanent focal ischemia. Journal of Cerebral Blood Flow & Metabolism, 2003. 23: p. 718-727.
120. Majda BT, M.B., Rixon N, Knuckey NW, Suppression subtraction hybridization and northern analysis reveal upregulation of heat shock, trkB, and sodium calcium exchanger genes following global cerebral ischemia in the rat. Molecular Brain Research, 2001. 93: p. 173-179.
121. Geddes JW, P.L., Holtz ML, Craddock SD, Maines MD., Permanent focal and transient global cerebral ischemia increase glial and neuronal expression of heme oxygenase-1, but not heme oxygenase-2, protein in rat brain. Neuroscience Letters, 1996. 210(3): p. 205-208.
122. . Matsuo K, T.S., Sekido H, Morita T, Kamiyama M, Morioka D, et al., Pharmacologic preconditioning effects: prostaglandin E1 induces heat-shock proteins immediately after ischemia/reperfusion of the mouse liver. Journal of Gastrointestinal Surgery, 2005. 9: p. 758-768.
123. Ren M, S.V., Chen RW, Chuang DM, Postinsult treatment with lithium reduces brain damage and facilitates neurological recovery in a rat ischemia/reperfusion model. Proceedings of the National Academy of Sciences of the United States of America, 2003. 100: p. 6210-6215.
124. DEMARCAY, H., Ueber die Natur der Galle. Annals of 1838. 27: p. 270-291.
125. Ilker Tasci, N.M., Mehmet Refik Mas, Murvet Tuncer, Bilgin Comert, Ultrastructural changes in hepatocytes after taurine treatment in CCl4 induced liver injury. World Journal of Gastroenterology, 2008. 14(31): p. 4897-4902.
126. Huxtable, R.J., Physiological Actions of Taurine. Physiological Reviews, 1992. 72(1): p. 101-163.
127. Pasantes-Morales, H., O. Quesada, L. Alcocer, and R. Sanchez-Olea, Taurine content in foods. Nutrition Report International, 1989. 40: p. 793-801.
128. Pasantes-Morales, H., R. Lopez-Escalera, and J. Moran, Taurine and zinc in nutrition and cellular development, in Current Topics in Nutrition and Disease (Basic and Clinical Aspects of Nutrition and Brain Development). 1987, Liss: New York. p. 217-243.
129. Davis, J.M.a.W.A.H., Amino acids and proteins of developing mammalian brain, in Biochemistry of the Devloping Brain, W.A. Himwich, Editor. 1973, Dekker: New York. p. 55-110.
130. PASANTES MORALES, H., Taurine function in excitable tissues: the retina as a model, in Retinal Transmitters and Modulators: Models for the Brain, W.W. Morgan, Editor. 1985: Boca Raton. p. 33-62.
131. Huxtable, R.J., Taurine in the central nervous system and the mammalian actions of taurine. Progress in Neurobiology, 1989. 32: p. 471-533.
132. PASANTES MORALES, H., Current concepts on the role of taurine in the retina. Progress in Retinal Research, 1986. 5: p. 207-230.
133. CHESNEY, R.W., A. L. Friedman, P.W. Albright, N. Gusowski, Fasting reverses the renal adaption to altered dietary sulfur amino acid intake. Proceedings of the Society for experimental Biology and Medicine, 1982. 170: p. 493-501.
134. CHESNEY, R.W., N. Gusowski, S. Dabbagh, M. Theissen, M. Padilla, and A. Diehl, Factors affecting the transport of β-amino acids in rat renal brush border membrane vesicles. Biochimica et Biophysica Acta, 1985. 812: p. 702-712.
135. Chesney, R.W., W. Jolly, I. Zelikovic, C. Iwahashi, and P. Lohstroh, Increased Na+-taurine symporter in rat renal brush border membranes: preformed or newly synthesised. FASEB Journal, 1989. 3: p. 2081-2085.
136. Huxtable, R.a.R.B., Taurine and isethionic acid: distribution and interconversion in the rat. The Journal of Nutrition, 1972. 102(7): p. 805-814.
137. Wheler, G.H.T., and D. C. Klein, Taurine release from the pineal gland is stimulated via a β-adrenergic mechanism. Brain Research, 1980. 187: p. 155-164.
138. Wheler, G.H.T., and D. C. Klein, Cyclic AMP-induced release of [14C]taurine from pinealocytes. Biochemical and Biophysical research communications, 1979. 90(1): p. 22-27.
139. Holopainen, I., P. Kontro, and S.S. Oja, Release of preloaded taurine and hypotaurine from astrocytes in primary culture: stimulation by calcium-free media. Neurochemical Resarch, 1985. 10: p. 123-132.
140. Madellain, V., D. L. Martin, R. Lepore, M. Perrone, and W. Shain, β-Receptor stimulated and cyclic adenosine 3',5' monophosphate-mediated taurine release from LRM55 glial cells. Journal of Neuroscience, 1985. 5: p. 3154-3160.
141. Thurston, J.H., R. E. Hauhart, and E.F. Naccarato, Taurine: possible role in osmotic regulation of mammalian heart. Science, 1981. 214: p. 1373-1374.
142. Thurston, J.H., R. E. Hauhart, and J. A. Dirgo, Taurine: a role in osmotic regulation of mammalian brain and possible clinical significance. Life Sciences, 1980. 26: p. 1561-1568.
143. Atlas, M., J. J. Bahl, W. Roeske, and R. Bressler, In vitro osmoregulation of taurine in fetal mouse hearts. Journal of molecular and cellular cardiology, 1984. 16: p. 311-320.
144. Jones, D.P., L. A. Miller, and R. W. Chesney, Adaptive regulation of taurine transport in two continuous renal epithelial cell lines. Kidney International, 1990. 38: p. 219-226.
145. Dantzler, W.H., and S. Silbernagl, Renal tubular reabsorption of taurine, GABA, and β-alanine studied by continous microperfusion. Pflugers Archiv European Journal of Pysiology, 1976. 367: p. 123-128.
146. HUXTABLE, R.J., AND L. A. SEBRING, Cardiovascular actions of taurine, in Sulfur Amino Acids: Biochemical and Clinical Aspects, R.J.H. K. Kuriyama, and H. Iwata, Editor. 1983: New York. p. 5-38.
147. FRANCONI, F., F. MARTINI, I. STENDARDI, R. MATUCCI, L. ZILLETTI, AND A. GIOTTI, Effect of taurine on calcium levels and contractility in guinea pig ventricular strips. Biochemical Pharmacology, 1982. 31(20): p. 3181-3186.
148. FRANCONI, F., I. STENDARDI, F. MARTINI, L. ZILLETTI, AND A. GIOTTI., Interaction between organic calcium-channel blockers and taurine in vitro and in vivo. The journal of Pharmacy and Pharmacology, 1982. 34: p. 329-330.
149. FRANCONI, F., I. STENDARDI, R. MATUCCI, P. FAILLI, F. BENNARDINI, G. ANTONINI, AND A. GIOTTI, Inotropic effect of taurine in guinea-pig ventricular strips. European Journal of Pharmacology, 1984. 102: p. 511-514.
150. KHATTER, J.C., P. L. SONI, R. J. HOESCHEN, L. E. ALTO, AND N. S. DHALLA., Subcellular effects of taurine on guinea pig heart., in The Effects of Taurine on Excitable Tissues, S.I.B. S. W. Schaffer, and J. J. Kocsis, Editor. 1981, Spectrum: New York. p. 281-294.
151. WELTY, J.D., AND W. 0. READ, Studies on the function of taurine in the heart. Proc. SD Acad. Sci., 1963. 17: p. 157-163.
152. Azari, J., P. Brumaugh, and R. J. Huxtable, Prophylaxis by taurine in the hearts of cardiomyopathic hamsters. Journal of molecular and cellular cardiology, 1980. 12: p. 1353-1366.
153. CHAZOV, E.I., L. S. MALCHIKOVA, N. V. LIPINA, G. B. ASAFOV, AND V. N. SMIRNOV, Taurine and electrical activity of the heart. Circulation Research. 35(Suppement 3 ): p. 11-21.
154. ITO, R., T. UCHIYAMA, S. YODA, N. HOMMA, AND K. FURUKAWA., Cardiovascular actions of taurine, γ-aminobutyric acid (GABA) and γ-amino-β-hydroxybutyric (GABOA) after chemical denervation., in The Effects of Taurine on Excitable Tissues, S.I.B. S. W. Schaffer, and J. J. Kocsis., Editor. 1981, Spectrum: New York. p. 313-327.
155. READ, W.O., AND J. D. WELTY, Effect of taurine on epinephrine and digoxin-induced irregularities of dog heart. Journal of Pharmacology And Experimental Therapeutics, 1963. 139(3): p. 283-289.
156. TAKAHASHI, K., J. AZUMA, N. AWATA, A. SAWAMURA, S. KISHIMOTO, T. YAMAGAMI, T. KISHI, H. HARADA, AND S. W. Schaffer, S. I. Baskin, and J. J. Kocsis, Protective effect of taurine on the irregular beating pattern of cultured myocardial cells induced by high and low extracellular calcium ion. Journal of molecular and cellular cardiology, 1988. 20(5): p. 397-403.
157. KRAMER, J.H., J. P. CHOVAN, AND S. W. SCHAFFER, Effect of taurine on calcium paradox and ischemic heart failure. Heart and Circulatory Physiology, 1981. 240: p. 238-246.
158. FRANCONI, F., I. STENDARDI, P. FAILLI, R. MATUCCI, C. BACCARO, L. MONTORSI, R. BANDINELLI, AND A. GIOTTI., The protective effects of taurine on hypoxia (performed in the absence of glucose) and on reoxygenation (in the presence of glucose) in guinea-pig heart. Biochemical Pharmacology, 1985. 34(15): p. 2611-2616.
159. KUO, C.H., AND N. MIKI., Stimulatory effect of taurine on Ca uptake by disc membranes from photoreceptor cell outer segments. Biochemical and Biophysical research communications, 1980. 94(2): p. 646-651.
160. Pasantes-Morales, H., Taurine-calcium interactions in frog rod outer segments: taurine effects on an ATP-dependent calcium translocation process. Vision research, 1982. 22: p. 1487-1493.
161. Harada, H., S. Allo, N. Viyuoh, J. Azuma, K. Takahashi, and S. W. Schaffer, Regulation of calcium transport in drug-induced taurine-depleted hearts. Biochimica et Biophysica Acta, 1988. 944: p. 273-278.
162. PASANTES-MORALES, H., AND C. CRUZ, Protective effect of taurine and zinc on peroxidation-induced damage in photoreceptor outer segments. Journal of Neuroscience Research, 1984. 11: p. 303-311.
163. PASANTES-MORALES, H., R. LOPEZ-ESCALERA, AND J. MORAN., Taurine and zinc in nutrition and cellular development., in Current Topics in Nutrition and Disease (Basic and Clinical Aspects of Nutrition and Brain Development), B.H. D. K. Rassin, and B. Drujan, Editor. 1987, Liss: New York. p. 217-243.
164. BUEGE, J.A., AND S. D. AUST., Microsomal lipid peroxidation. Methods in enzymology, 1978. 52: p. 302-310.
165. FELLMAN, J.H., AND E. S. ROTH, The biological oxidation of hypotaurine to taurine: hypotaurine as an antioxidant., in Taurine: Biological Actions and Clinical Perspectives, L.A. S. S. Oja, P. Kontro, and M. K. Paasonen, Editor. 1985, Liss: New York. p. 71-82.
166. Aruoma, O.I., B. Halliwell, B. M. Hoey, and J. Butler, The antioxidant action of taurine, hypotaurine and their metabolic precursors. Biochemical Journal, 1988. 256: p. 251-255.








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Comments (11)


Fail day is fail
  PokerDoc88, Nov 19 2008

Today has been full of fail.

I am going to go all emo, so bear with me. First of all, I had 4 hours of sleep. I had a dental appointment at 8:30, and normally it takes 30 minutes to drive there. I left at 7:45 anyway just to make sure I'm not late, becoz last time I was late the dentist got really fkn angry at me and forced me to make an additional appointment. I end up being late ANYWAY because traffic fucking sucked and really retarded shit happened like a semi-trailer blocking an intersection totally for 4 minutes.

Dental appointment was shit (standard). Afterwards I have to go to the shithole lab where I'm stuck for a year being as a research-student gimp who is essentially slave labor (I don't get paid for a single thing I do there, and my boss constant makes me do things that aren't 'required' of me becoz hes too fucking cheap to employ an actual scientist).

First I drop a box of frozen antibody tubes all over the floor. That's not such a bad fail. Later on I was emptying used jars of xylene (a toxic chemical) for disposal, and the bottle I'm pouring into begins to overflow (I can't see the level rising coz the bottle is really dark brown glass). So my supervisor gets all pissed off at me becoz of this huge spill of toxic shit and thinks I'm a retard. Then the lab work I'm doing fails anyway, becoz the slides dried out hence ruining the staining procedure. Bye bye 2 days of work. Basically my entire day at the lab was fail.

Oh well, I'll go home and life will be EZ. HO HO think again sucker. It's raining like crazy so I have to run to my car. My demister isn't working so as I drive home, I'm constant wiping the windshield with my sleeve so I can see where I'm going. Then the biggest fail of all (and not even my fault): as I'm driving along, a semitrailer comes to merge into my lane to do a left-hand turn, but it doesn't see me. Result: I have to slam the breaks to avoid getting crushed, and the back of the truck still hits the front of my car anyway. Fucking retarded son of a bitch.

So I continue driving, and then 5 minutes later another suicidal fucking spastic driver pulls out infront of me from a park spot and forces me to slam the breaks YET AGAIN. So WTF I'm just fucking hungry, I haven't eaten a single bit of food all day and its 3pm, so I go to Maccas to get Big Mac Meal with BBQ sauce (Win). Unfortunately, stupid bitch forgets to put sauce in the bag (fail). I'm so pissed off at this stage that I just parked and walked in, asking for sauce. Normally when they forget I don't care, but goddamn it I'm getting my fukn sauce one time! Thankfully the manager was really nice and gave me 2 sauces (victory of the day).

I finally get home with my food and change into my PJ's (yay). Strangely no one seems to be around, which is rare because my parents are retired and one of them is basically always home. So I go look for my parents and then wtf happens? I walk in on them having sex. Seriously what the fucking hell. I have not walked in on them for years and years and years. I didn't even think they had sex any more. Seriously what the fucking hell fuck. I just want to say hi to them and get some steam off my chest. But noooo, I open the door and I hear "HANG ON!" and then "OH I HAVE A CRAMP". Seriously WHAT THE FUCK. Jesus fucking christ. So tilted.

I guarantee you if I played poker right now, I'd get dealt KK vs AA twice and lose, then AA vs KK and get sucked out on. That's how this day feels.

/emo
/rant

so sup LP?



*****1 votes

Comments (7)


"I'm going to post this on 2+2!!!:
  PokerDoc88, Nov 10 2008

Submitted by : PokerDoc88

PokerStars Game #21912214582: Hold'em No Limit ($0.10/$0.25) - 2008/11/10 3:12:14 ET
Table 'Sycorax III' 9-max Seat #9 is the button
Seat 1: goldenglove2 ($58.35 in chips)
Seat 2: JCesana3 ($22.30 in chips)
Seat 3: Fish4luck ($31.75 in chips)
Seat 4: porkygirl ($11.25 in chips)
Seat 5: Dominicjs1 ($12.10 in chips)
Seat 6: Xisco ($29.95 in chips)
Seat 7: LilClinkin ($24.95 in chips)
Seat 8: enright3060 ($23.75 in chips)
Seat 9: Queenod ($22.90 in chips)
goldenglove2: posts small blind $0.10
JCesana3: posts big blind $0.25

Holecards(Odds)
Dealt to LilClinkin AsKd
Fish4luck: folds
porkygirl: folds
Dominicjs1: folds
Xisco: folds
LilClinkin : raises $0.75 to $1
enright3060: folds
Queenod: folds
goldenglove2 said, "dominic james?"
goldenglove2: calls $0.90
JCesana3: folds

Flop(Odds) (Pot : $2.25)

   9s9d8s
goldenglove2: checks
LilClinkin : checks

Turn(Odds) (Pot : $2.25)

   9s9d8s3d
goldenglove2: bets $1
LilClinkin : calls $1

River (Pot : $4.25)

   9s9d8s3dJd
goldenglove2: bets $2.50
LilClinkin : calls $2.50

Showdown
goldenglove2: shows AdTc (a pair of Nines)
LilClinkin : shows AsKd (a pair of Nines - Ace+King kicker)
LilClinkin collected $8.80 from pot

Summary
Total pot $9.25 | Rake $0.45
Board  9s9d8s3dJd
Seat 1: goldenglove2 (small blind) showed AdTc and lost with a pair of Nines
Seat 2: JCesana3 (big blind) folded before Flop
Seat 3: Fish4luck folded before Flop (didn't bet)
Seat 4: porkygirl folded before Flop (didn't bet)
Seat 5: Dominicjs1 folded before Flop (didn't bet)
Seat 6: Xisco folded before Flop (didn't bet)
Seat 7: LilClinkin showed AsKd and won ($8.80) with a pair of Nines
Seat 8: enright3060 folded before Flop (didn't bet)
Seat 9: Queenod (button) folded before Flop (didn't bet)



Dealer: Game #21912214582: LilClinkin wins pot ($8.80) with a pair of Nines
goldenglove2: hahaha
goldenglove2: i like you
goldenglove2: pls make that call all the time
goldenglove2: ty
LilClinkin : plz donk like that
LilClinkin : ty
goldenglove2: hahaha you call with A high on that board
goldenglove2: i like you
LilClinkin : stop trying to make your self ownage look like a victory
goldenglove2: oooooo but it was AKKKKKKKK
Dealer: goldenglove2, it's your turn. You have 8 seconds to act
Dealer: Game #21912224974: Fish4luck wins pot ($0.95)
goldenglove2: that 9$ pot was total self ownage
goldenglove2: im going to go put this on 2+2 to make fun of you
LilClinkin : don't go getting all twisted up about it
Dealer: goldenglove2, it's your turn. You have 8 seconds to act
LilClinkin : ok hf sir



LOL



0 votes

Comments (10)


lol
  PokerDoc88, Oct 06 2008

lol



0 votes

Comments (3)


Bohemian grove: World leaders satanic cultists?
  PokerDoc88, Aug 10 2008

Hey lately I've been doing a lot of looking into of conspiracy theories after seeing Zeitgeist. While some of what you see in Zeitgeist is obv bullshit, there is definitely some truth to it. The key message is to question everything, seek knowledge, and make up your own mind.

This is a series of 7 videos I found on youtube about a place called "Bohemian Grove". Watch and make up your own mind.





0 votes

Comments (0)


Return to poker, and why Creationists are retards
  PokerDoc88, Jul 05 2008

Ok so for the past 4 months or so, I haven't touched the game. I decided I'd focus more on uni and take a break from poker. Exams finished 3 weeks ago, results came out (I did the same as I've done previous semesters =/ ) and since then I've returned to playing.

Deciding that FTP really is RIGGD, I decided to put $500 on stars and grind up from 25nl to ease my way back in.

I've noticed the games on stars are fairly soft, there are quite a few fish but then there are also quite a few multi-tabling nitty regs. Not only that, but most of the fish don't really get involved in big pots without the nuts. So, while its easy to make money, it takes a lot of diligent grinding and disciplined play to do so.

Here's the graph of my run at 25nl: (Note the first 6k hands I donked around really bad, playing like 28/26 and trying to run stations over...obv failed haha).



So I'll continue grinding, moving up to 50nl next session I play. Holidays only last for 1 more week, but I don't return to uni: I'm doing research in a lab at a hospital for a year. Hopefully it's not too full-on, and I won't have any commitment to it at home, so I can continue playing poker for the coming year.

In non-poker related stuff, I have a long to-do list of games to complete:

On XBox360 I have to finish Lost Odyssey, GTA4, DMD mode on DMC4, Ninja Gaiden 2 (this is fucking hard) and Gears of War.

Also, I'm going to go out and buy a parts for a new computer in a few days. While not finalized, the setup seems to be:

Core2Duo 8500
ATI Radeon 4850/4870 (undecided on which card to get yet)
4gig DDR2 ram
Appropriate case and PSU
22" LCD

My friend made a detailed list of the exact parts and costs, it should be$1.2 - $1.4k Australian dollars. And ready to kick ass in SC2 at max settings (hopefully) when it finally comes out.

Finally, some funny youtube videos which bag creationists (if you are a creationist, lol @ you)







There's over 20 videos in this series, but these 3 are amongst the funniest. I suggest you listen while you grind, it's good entertainment and tilt relief (unless of course you're a creationist, but in that case you probably think poker is all about luck and suck).





0 votes

Comments (1)


Surgery happened! Feel shitty
  PokerDoc88, Feb 02 2008

So I had my jaw surgery yesterday, it was 3.5 hour operation. Thankfully, the surgeon only had to adjust my upper jaw, so my lower jaw remains unscathed.

I still feel the effects of the general anesthetic, although it is very mild now. The nerve blocks they used during the surgery are wearing off, so I am beginning to feel dull throbbing pain. However, it is far less severe than I imagined it would be, so I'm very relieved. My face is massively swollen, I'll post photos of myself later if I can be bothered. I'm very tired and have a headache. The most frustrating thing is not being able to eat anything except for complete mush, and I have frequent nose bleeds. I'm told these will go away soon, and within a week I should be able to chew soft foods.

I'm very tired, but I can't sleep properly because I favor positions where I have my face buried in the pillow: Obviously that's not possible because it is painful to touch my face around the mouth area, so I have to get by on fragmented sleep on my back .

Here's to hoping for a speedy recovery!



0 votes

Comments (7)


Jaw surgery soon
  PokerDoc88, Jan 25 2008

So I got jaw surgery coming up, I'm booked in Saturday morning on the 2nd of Feb. Because I consider this to be fairly major event in my life, and because I want a quick and complication-free healing period afterward, I'm deciding to not play any more poker until the surgery is done and probably won't play for at least 2 weeks afterwards. The reason for not playing prior to the surgery is, it would be really gay to go on a 10bi downswing, mentally feel like shit, and then go under the knife. I'm happy with my poker progress this month so far (+$2k in approx 35k hands), so I'll just let it stay that way.

The purpose for the surgery is I have a very big over-bite, meaning my top teeth protrude over my bottom teeth. Because of this, when I bite down fully, my teeth only touch at the back molars, and the front teeth make no contact whatsoever. The gap at the front is so large that I can bite down fully, and still poke my tongue through with ease.

These pictures kinda show what is being done, I'll try to commentate exactly what each picture is trying to show.

-This shows a side-on view of the skull only. The black line shows the cut they will be making into my maxilla to adjust my upper jaw. The result is to bring my upper teeth lower and backwards.

http://www.dropshots.com/LilClinkin#date/2008-01-25/18:10:21

-This shows a frontal view of both the skull and mandible. The cut-lines are in red this time, and show how they are cutting directly back into my maxilla (upper jaw), and how they are cutting my lower jaw on both sides to adjust the angle, bringing it upwards to cause my lower teeth to move higher and more forward.

http://www.dropshots.com/LilClinkin#date/2008-01-25/18:15:32

-This shows a top-down view of the mandible (lower jaw). It's hard to explain exactly how the cut looks, but it is basically a really shallow-angled cut through the bone on each side. The red line shows the cut-line they will use, and the green shows the surface area they have of each piece of bone on which to manipulate it and adjust the angle. The goal is to tilt the bone upwards, so that my lower teeth will come up and forward.

http://www.dropshots.com/LilClinkin#date/2008-01-25/19:05:05

I'm getting kind of nervous because post-op is going to suck and be painful. Apparently it's standard to lose 5kg post-op, due to the trauma of the surgery and the fact I can only eat limited amounts of soft food afterwards.

edit. The pics don't work directly, so you'll just have to make do with links.



0 votes

Comments (6)


BR hit 4k
  PokerDoc88, Jan 20 2008

Yeah, played 1k session at 100nl today, up about 1.5 buyins which puts my br over 4k!!! I shouldn't be too proud of the wins though, they were both suck-outs. Not 'donkey' suckouts, but suckouts nonetheless (ie. I float and end up 2-outering with a low pp vs AA, where normally I would fold to a 2nd barrel).



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Comments (6)




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