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===Mitochondria-mediated pathway===
===Mitochondria-mediated pathway===
Mitochondria-mediated pathways are an intracellular activated pathway, which is activated from the release of cytochrome c as a result of increased membrane permeability of the mitochondria itself (1). The release of cytochrome c occurs from both the inner mitochondrial membrane and the outer mitochondrial membrane (3). The release of cytochrome c into the cytosol results in the formation of apoptosomes. These wheel-like complexes are formed by the binding of apoptotic protease activating factor 1 [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=317&ordinalpos=2&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum Apaf-1] and [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=842&ordinalpos=6&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum caspase-9] (1). Conformational changes occur within the complex, which cleaves and in turn activates caspase-9, which then activates caspase-3. Caspase-3 will cleave the target proteins, initiating cellular degradation.
Mitochondria-mediated pathways are an intracellular activated pathway, which is activated from the release of cytochrome c as a result of increased membrane permeability of the mitochondria itself (1). The release of cytochrome c occurs from both the inner mitochondrial membrane and the outer mitochondrial membrane (3). The release of cytochrome c into the cytosol results in the formation of apoptosomes. These wheel-like complexes are formed by the binding of apoptotic protease activating factor 1 [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=317&ordinalpos=2&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum Apaf-1] and [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=842&ordinalpos=6&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum caspase-9] (1). Conformational changes occur within the complex, which cleaves and in turn activates caspase-9, which then activates caspase-3. Caspase-3 will cleave the target proteins, initiating cellular degradation.
===ER-mediated pathway===
===ER-mediated pathway===
Revision as of 13:24, 27 May 2009
- 1 Introduction
- 2 Definition
- 3 What Happens During Apoptosis?
- 4 Why Does Apoptosis Occur?
- 5 Brief History Of Apoptosis
- 6 References
- 7 Signaling
- 8 Apoptosis Signalling Diagram
- 9 Regulations of the apoptosis
- 10 The morphology of apoptosis
- 11 Timeline
- 12 Design model of apoptosis
- 13 Apoptosis from a Clinical Point of View
- 14 References
- 15 Glossary
- 16 2009 Group Projects
In all living organisms there is a harmonious balance between the synthesis of new cells and the destruction of existing cells under normal conditions. This balance is particularly vital in maintaining both proper function and structure within the organism. The complex events that encompass these processes take place in response to specific and controlled signals. In this paper, focus is placed upon the process of "Programmed Cell Death" (PCD), more commonly known as Apoptosis, and the significance of this process in maintaining cell population. Mechanisms, abnormalities and future implications of research in Apoptotic Cell Death will also be explored.
The word Apoptosis is derived from a word of ancient Greek origin: απóπóτσισ. This literally means "falling off" or "falling away" as in leaves from a tree in autumn[1] which incidentally also involves apoptosis. During Apoptosis, cells are described as "programmed" to self-destruct or commit "cellular suicide", as opposed to "cellular homicide", characteristic of Necrotic Cell Death (another form of cell death)[2]. The term Apoptosis was first used by John Kerr[3] in 1972[4], in reference to the morphological presentation of ''apoptotic bodies'', characteristic of what Glucksmann[5] described as developmental cell death in 1951[4]. Years of research in cell biology have defined apoptosis as a physiological process that occurs via an extrinsic or intrinsic (mitochondrial) pathway. Both of these pathways involve activation of the Caspase Cascade, which eventuates in the death of the cell. This process is an essential part of normal physiological function and as a result each day billions of cells in the human body die via apoptosis, particularly blood cells and those in epithelia lining organs such as the intestine[6].
What Happens During Apoptosis?
This process of Apoptosis requires specific chemical and biological conditions that activate what is known as the Caspase Cascade. Specialized proteins including Bax and Bcl-2 are the primary intracellular signaling proteins involved in activating the Caspase Cascade that induces apoptosis[7]. The various biochemical mechanisms that characterize the Caspase Cascade ultimately eventuate in death of the cell. Apoptosis is an ATP-dependant intracellular process that may be induced by numerous different stimuli including infectious and injurious agents, for example viruses and mutagenic agents respectively. However it is important to note that not all cells present the same sensitivity to stimuli. Upon receiving signals from such stimuli the cell self-instructs itself to die[2]. Apoptosis includes a variety of highly orchestrated events; some that work in parallel with others and some that work in sequence to others. These events will be discussed in detail below, however in brief they include the initiation of the caspase cascade, cell shrinkage, protein fragmentation, chromatin condensation, DNA degradation and plasma membrane blebbing, finally followed by rapid engulfment of the deceased cell by neighbouring phagocytic cells[8].
Why Does Apoptosis Occur?
Apoptosis occurs as a normal process in most living organisms, including humans. In humans it is an integral component of normal development of the embryo, maturation and cell turnover and has also been implicated in a range of pathologically significant conditions, particularly cancer and autoimmune disease[9]. Apoptosis most commonly occurs as a homeostatic process which is carried out by cells of living organisms as a means of eliminating unwanted cells and to maintain cell population in tissue. Unwanted cells can be described as those that have been exposed to injurious or infectious agents rendering them damaged and potentially dangerous to the rest of the organism[9]. This defense mechanism can occur as a response to viral or bacterial infection, damage to DNA, for example by mutagenic agents, genetic disorders, as in autoimmune disease and various other injurious agents. A vast amount of apoptosis can occur during the developmental stages of an organism. A common example is the formation of the human hand during embryological development. In its initial stages the hand begins as a paddle-like structure, however in order for digitization to occur the inter-digital cells must undergo apoptosis resulting in the formation of a normal human hand[10]. Another frequent example is the development of a tadpole into a frog. In the immune system cells infected by a virus may recognise the infectious agent triggering the process of apoptosis. Apoptosis acts to destroy the cell to prevent the virus from replicating and infecting further cells of the organism[10]. Although apoptosis occurs under normal physiological conditions it may also occur pathologically in an abnormal disease state. Apoptosis has been implicated in many chronic disease processes such as those present in autoimmune diseases, for example rheumatoid arthritis, and neurodegenerative diseases including Multiple Sclerosis and Parkinson's disease which is thought to be associated with too much apoptosis[1]. On the contrary, too little apoptosis is of particular significance in the formation of cancer, essentially due to mutations in cancer cells that prevent them from undergoing apoptosis. Apoptosis has also been observed in plants, particularly higher plants. The xylem in plants consists of spaces that result from the death of cells that previously occupied those spaces[10], comparable to the digitization of the hand during embryological development.
Brief History Of Apoptosis
- 1842: Carl Vogt described that cells die during normal vertebrate development[11].
- 1951: Glucksmann wrote an influential review on apoptosis and re-sparked early interest in the field of apoptosis research[4].
- 1972: John Kerr first coined the term Apoptosis. John Kerr. He noticed apoptosis in hepatocytes but had originally termed this process "shrinkage necrosis" due to the observed shrinkage of the dying hepatocytes. Further to this, Kerr recognized that unlike necrosis, no inflammation was present hypothesised that the process being observed was separate from necrosis. He then helped confirm this hypothesis using electron microscopy to identify and describe morphological differences between necrosis and apoptosis[1][4]
- 1976: Programmed Cell Death was first observed in the nematode Caenorhabditis elegans by J.E. Sulston and described as a "defined pattern of cell deaths"[12].
- 1980: Wyllie discovered that DNA degredation and nuclear chromatin condensation was characteristic of apoptosis and that these events where associated with intracellular, endogenous signals[13].
- 1982 - 1986: Horvitz et al.[14] discovered that cell death occurred as a natural process during development of the nematode Caenorhabditis elegans. It was discovered that the worm began with 1090 somatic cells before reaching a mature state after which 131 died through apoptosis, leaving the hermaphrodite worm with a total of 959 somatic cells[15]. Identification of genetic pathways dedicated to this programmed death of cells and the production the first cell death mutants (ced-1, ced-2) occurred soon after[16][17][4]. These were milestone discoveries that sparked unparalleled interest in the field of apoptosis.
- 1988: Vaux et al.[18] first identified a component of the apoptosis mechanism - Bcl-2; an important cell death inhibitor.
- 1991: Yonish et al.[19] determined that p53 was able to induce apoptosis (in myeloid leukaemic cells).
- 1994: Birnbaum et al. identified an apoptosis inhibiting gene[20] which in turn lead to the identification of IAPs and their specific roles[4].
- 1994 - Current: many researchers identified genes and proteins involved in the induction and inhibition of apoptosis, regulation of the process and the many receptors and pathways involved. Some of these important apoptotic factors include IAPs (Inhibitor Apoptotic Proteins), TNF (tumor necrosis factor), caspases (intracellular proteolytic enzymes) and bax (pro-apoptotic intracellular enzyme).
- Current: Much research is currently being conducted in the area of Apoptosis, however our aim to better understand and determine the process and factors involved, is being combined with the modern focus directed towards the therapeutic applications of our knowledge of Apoptosis, particularly factors that induce and inhibit the process and how these may be implicated in pathologies such as cancer, AIDS, autoimmune disease and neurodegenerative disease[21]. Although copious amounts of information have been found in relation to apoptotic cell death, it has been established through many unanswered questions in the area that further research is highly warranted[22].
[1] Lawen A. (2003). BioEssays. Apoptosis - an introduction, 25:888–896.
[2] Vinay Kumar et al. (2008) Robbins Basic Pathology. (8th Edition). Philadelphia, Pa.: Saunders/Elsevier.
[3] Kerr JFR, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972;26:239–571.
[4] Vaux DL. (2002). Cell Death Differentiation. Apoptosis timeline, 9:349–354.
[5] Glucksmann A (1951) Cell deaths in normal vertebrate ontogeny. Biol. Rev. 26: 59±86.
[6] William K. Purves et al. (2004) Life - The Science of Biology. (7th Edition). Gordonsville, VA 22942 U.S.A.: VHPS/W.H. Freeman & Co.
[7] Bruce Alberts et al. (2002). Molecular Biology of the Cell. (5th Edition). New York, USA.: Garland Science.
[8] Kimball's Biology Pages (2008). Apoptosis. Retrieved on May 20th, 2009 from source http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/Apoptosis.html
[9] Elmore S. (2007). Toxicologic Pathology. Apoptosis: A Review of Programmed Cell Death, 35:495-516.
[10] Robert H. Horvitz (1999). Why Does Programmed Cell Death or Apoptosis Occur? Retrieved on May 16th, 2009 from source http://www.scientificamerican.com/article.cfm?id=why-does-programmed-cell
[11] Peter ME, Heufelder AE, Hengartner MO. (1997). Proceedings of the National Academy of Science of the United States of America. Advances in apoptosis research, 94(24):12736-7.
[12] Sulston JE. (1976). Philosophical Translations of the Royal Society of London. Series B, Biological Sciences. Post-embryonic development in the ventral cord of Caenorhabditis elegans, 275(938):287-97.
[13] Wyllie AH. (1980). Nature. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation, 284(5756):555-6.
[14] Horvitz HR, Ellis HM and Sternberg PW (1982) Programmed cell death in nematode development. Neurosci. Comment. 1: 56 ± 65.
[15] Hengartner MO, Horvitz HR. (1994). Philosophical Translations of the Royal Society of London. Series B, Biological Sciences. The ins and outs of programmed cell death during C. elegans development, 345(1313):243-6
[16] Ellis HM, Horvitz HR. (1986). Cell. Genetic control of programmed cell death in the nematode C. elegans. 44(6):817-29.
[17] Horvitz RH. (1999). Cancer Research. Genetic Control of Programmed Cell Death in the Nematode Caenorhabditis elegans, 59, 1701s-1706s.
[18] Vaux DL, Cory S and Adams JM (1988). Nature. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells, 355: 440 ± 442.
[19] Yonish RE, Resnitzky D, Lotem J, Sachs L, Kimchi A and Oren M. (1991). Nature. Wildtype p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6, 353: 345 ± 347.
[20] Birnbaum MJ, Clem RJ and Miller LK. (1994). Journal of Virology. An apoptosis inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with Cys/His sequence motoif, 68: 2521 ± 2528.
[21] Diamantisa A, Magiorkinisa E, Sakorafasb GH, Androutsosc G. (2008). Onkologie. A Brief History of Apoptosis: From Ancient to Modern Times, 31:702–706.
[22] Formigli L, Conti A, Lippi D. (2004). Minerva Medica. "Falling leaves": a survey of the history of apoptosis, 95(2):159-64.
Apoptosis signalling is a multi-pathway mechanism that results in the convergence of the activation of caspases (a family of cystiene proteases). These pathways are activated by either extracellular or intracellular stimuli. The caspases are activated by four major pathways; these include Mitochondrial-mediated pathway, death receptor-mediated pathway, granzyme-A and granzyme-B-mediated pathway, and Endoplasmic Reticulum (ER) - mediated pathway (1).
Death receptor-mediated pathway
Death receptor-mediated pathways are an extracellular activated pathway. The death receptors Fas and the tumour necrosis factor receptor TNFR are found on the surface of the plasma membrane. They are activated by death ligands. The death ligands Fas Ligand FasL and tumour necrosis factor alpha TNF-α bind to the receptor – initiating the cascade. Death receptors are single transmembrane-spanning proteins; Fas and TNFR-1 receptors have a cytoplasmic tail which is known as the death domain (DD) (fig 1) (3). After binding, the death receptors undergo trimerisation, which signals for the death inducing signalling complex (DISC). It consists of the recruitment of adaptor proteins, procaspase-8, and either Fas associated death domains (FADD) or TNF receptor associated death domain (TNFADD). Procaspase-8 cleavage of the pro-domain activates caspases-8 to its pro-apoptotic form (1), which then activates Caspase-3 (4). Caspases-3 protein can be activated in the death receptor-mediated pathway or the mitochondria-mediated pathway, and initiates DNA cleavage when activated. Caspase-3 cleaves a series of proteins responsible for maintaining cell life and structure. Including DNA metabolism substrates, such as poly(ADP-ribosome)polymerase PARP which facilitates DNA repair, cytoskeleton substrates, nuclear lamins, gelsolin, and fodrin (2,4). Caspase activation can be cross-linked, which demonstrates the signalling complexities of apoptosis. Caspase-8 can also act on the mitochondria-mediated pathway – amplifying the cascade of reactions (4). Bid is a proapoptotic member of the Bcl-2 family (1) that is activated by Caspase-8. It is located within the cytosol and, in the presence of caspase-8, can cleave to form tBid (truncated Bid). tBid can translocate to mitochondria – activating Bax, effects the functioning on the mitochondria and results in cytochrome c release (1).
Mitochondria-mediated pathway
Mitochondria-mediated pathways are an intracellular activated pathway, which is activated from the release of cytochrome c as a result of increased membrane permeability of the mitochondria itself (1). The release of cytochrome c occurs from both the inner mitochondrial membrane and the outer mitochondrial membrane (3). The release of cytochrome c into the cytosol results in the formation of apoptosomes. These wheel-like complexes are formed by the binding of apoptotic protease activating factor 1 Apaf-1 and caspase-9 (1). Conformational changes occur within the complex, which cleaves and in turn activates caspase-9, which then activates caspase-3. Caspase-3 will cleave the target proteins, initiating cellular degradation.
ER-mediated pathway
ER-mediated pathways have recently been linked to caspase activation. It is thought that the members of the Bcl-2 family that either inhibit or enhance apoptosis bind to the membrane of the ER (1). The response to abnormal events within the ER, such as a build up of misfolded proteins or the alteration in the homeostasis of calcium signalling, results in stress on the ER (3), which has been shown to initiate signalling for apoptosis. Caspase-12 is said to be activated by these stresses, and if over expression occurs, it can lead to the activation of other pathways (3). These pathways are not fully understood, but there are suggestions that activation of procaspase-9 and the formation of apoptosomes could be avoided via these pathways – directly activating mitochondria-mediated cytochrome c release via the activation of the Bax (3).
Granzyme –A and –B mediated pathways
Granzyme –A and –B mediated pathways are intracellular mediated pathways that release granzyme A (Grz A) and granzyme B (Grz B). Granzymes are serine proteases, which act very similarly to caspases. These pathways can be either independent or caspase-dependent pathways (1). Grz A is a slow acting independent pathway, which activates an ER associated complex known as SET. As a result of the reduction of reactive oxygen species (ROS) and mitochondrial membrane, signalling for the translocation of the complex into the cell's nucleus is induced. Once the complex is within the cell's nucleus, Grz A cleaves the complex – degrading the chromosomal DNA (4). Grz B is a depended-caspase pathway and is released from cytotoxic T Lymphocytes (CTL). It binds to cell surface receptors before being endocytosed into the cytosol with the help of a pore-forming protein perforin (1). Grz B triggers apoptosis in two ways; by cleaving Bid or by directly activating caspase-3. Grz B truncates Bid to tBid, which increases the membrane permeability of the mitochondria, consequently releasing cytochrome c. Cytochrome c actives caspase-9 (as described above) and downstream activation of caspase-3 occurs (4).
The mechanisms for apoptosis signalling are complex, and a lot is still not fully understood. There are a number of pathways which cross link, which enable the amplification of the apoptosis cascade. The diagram below demonstrates this.
Apoptosis Signalling Diagram
1. Wang ZB, Liu YQ, Cui YF. (2005). Pathways to caspase activation. Cell Bio Int. 29:489-496.
2. Stasser A, O'Connor Liam, and Dixit VM. (2000). Apoptosis Signalling. Annu.Rev.Biochem. 69:217-245
3. Czerski L, Nunez G. (2004).Apoptosome formation and caspase activation: is it different in the heart?. J Mol Cell Cardio. 37:643-652.
4. Chávez-Galán L, Arenas-Del Angel MC, Zenteno E, Chávez R and Lascurain R. (2009). Cell Death Mechanisms Induced by Cytotoxic Lymphocytes. Cellular & Molecular Immunology. 6(1):15-25.
Regulations of the apoptosis
Since apoptosis is leading to the irreversable damage of the cell due to the spontaneously activated caspases, it is crucial that caspase activations are tightly regulated. These regulation may delay or stop the apoptosis process so that it gives cell a time to repair the damage and return to their normal state. This is achieved by the adjustment of the availability of key anti- and pro- apoptotic molecules.[1][2]: Regulation of apoptosis mechanisms are complex and still in progress of research, however following topics are the major regulators of the apoptosis.
Bcl-2 family
Bcl-2 is the major regulator protein family of apoptosis via the mitochondrial mediated apoptosis pathway.There are around 25 different family members known to date, and both proapoptotic and antiapoptotic regulating proteins exist in this family. The activity and overall levels of pro- and anti-apoptotic families are under tight transcriptional and post-translational control. In healthy cells anti-apoptotic protein levels are high, whereas pro-apoptotic proteins are virtually absent or conformationally restrained. However, upon exposure to death signal this balance shifts in favour of pro-apoptotic proteins that are rapidly induced or dislodged. [1]
Antiapoptotic family
- Bcl-2, Bcl-XL, Bcl-W, Mcl-1 and Al/Bft-1 are major antiapoptotic proteins of the Bcl-2 family.
- Bcl-2 antiapoptotic proteins prevent mitochondrial changes; antiapoptotic family directly binding to and neutralizing the killer proteins(Bad), thus guard the mitochondrial membrane from pore formation. Formation of the pore may lead to release of cytochrome c and initiation of apoptosis.
- Bcl-2 family proteins(Bcl-XL,Bcl-2) can bind to the CED homolog Apaf-1 preventing its interaction with cytochrome c, thus preventing apoptosis.
- Antiapoptotic family can arrest the cell cycle mainly in G0 and G1 phase,it gives cell a chance to repaire the damage.[1][3]
Proapoptotic family
- Proteins such as Bax, Bak, and Mtd/Bok are the main proteins of proapoptotic family which have autonomous cytodestructive activity.
- They are able to bind Bcl-2, Bcl-XL, and prevent their antiapoptotic activity.
- They have similar structural to the pore-forming proteins. It enables them to form channels in the intracellular membranes and induce the release of cytochrome c from mitochondria.
- The induction of proapoptotic family protein is initiated after death stimuli. And p53 protein is also one of the initiator[3]
ADEDs family proteins
In the death receptor mediated pathway, adaptor protein interacts with death receptor and initiate apoptosis. One of ADEDs(Death effector domain proteins) proteins, cFlip, is a homolog of caspases-8 and -10 that contains DEDs but lacks proteolytic activity. cFlip and other antiapoptotic DED family proteins compete with caspases for binding to Fadd/Mort-1, thus functioning as a transdominant inhibitors of these caspases involved in TNF family cytokine signaling. [1]
IAPs family
Inhibitor of apoptosis protein(IAP) family proteins constitute a novel group of apoptosiss suppressors that are conserved throughout animal evolution. Although the mechanisms of these proteins suppress cell death are not well understood, the only clearly identified activity for these proteins is as inhibitors of caspases. Specifically, several of the human IAP family proteins have been reported to directly bind and potently inhibit particular members of the caspase family of cell death proteases.[4]:
Interestingly, not all caspases are targets of IAP suppression. To date, only caspases-3, -7 and -9 have been reported to be bound and inhhibited by IAPs. Caspases -1, -6, -8, and -10 are not. The significance of this selective inhibition of caspases by IAPs is that the inhibited caspases operate in the distal portions of the apoptotic proteolytic cascades, with some (such as caspases-3 and -7) functiooning as the ultimate effectors of apoptosis that cleave the various proteins responsible for cell death.[4]
Ubiquitin-mediated regulation
Ubiquitin is the protein which is often regarded as a tag for the 'waste product', it is now recognized it influences multiple biological processes, and it is also involved in regulation of the apoptosis. Ubiquitin is conjugated onto the protein and it directly influence the protein function. Moreover ubiquilated protein is recognised by Ub receptors and it leads to either degradation or culminating of the protein. With this mechanisms
- They control levels of many pro- and anti-apoptotic molecules
- induce apoptosis by promoting IAPs family degradation
- Ubiquitin may also limit the activation of caspases via ubiquitin (Ub)-dependent modifications.[5]
Guardian of the genome 'p53' maintains the genetic integrity by recognition of DNA damage and regulation of the apoptosis. It found very low amount in normal cell. Following the DNA damages and death stimuli, it is activated and performs its action.[6]
- When the damage is reversible, it arrests cell cycle and gives cell a time to repair.
- However if the damage is irreversible, it initiate the apoptosis through a mitochondrial pathway. Upon its activation, p53 binds to the outer mitochondrial membrane and induces permeabilization. It results release of cytochrome c which lead to caspase activation.[6]
Click here for a general overview of the p53 genome.
The morphology of apoptosis
The morphogenesis of apoptosis are characterised within the nucleus of the cell's compartment – in which, it illustrates four cardinal traits: volume reduction, chromatin condensation, recognition by phagocytic cells and phagocytosis.[7]
- Cell shrinkage
Osmosis is considered to be the offset in the commencement of the apoptotic pathway, whereby, due to the influx of sodium ions (Na+)
there is lost of intracellular fluid selectively.
Therefore, it illustrates in an abrupt onset of cellular shrinkage and leads to the induction of:
- Increase of the cell's density, in which, leads to the compression of the cytoplasmic organelles - the endoplasmic reticulum becomes dilated and the convolution of the cell and nuclear outlines becomes evident.
- Deterioration of the cell's specialised membrane integrity, such as, the loss of microvilli, junctional complexes or desmosomes.
- Lost of extracellular signaling between adjacent cells, as a result, it dissociates from neighboring cells.
This process allows the cell to reorganise its peripheral focal adhesions, in which, is also allows the cellular structure to adopt a more rounded morphological appearance.[7][8][9][10]
- Lost of plasma membrane integrity
The outer structural appearance of the cell formulates protrusions from the surface of its plasma membrane - known as blebs - the formation of blebs are due to the separation of the plasma membrane from the cytoskeleton of the cell.
The process of blebbing continues until the final molecular event of pyknosis – the condensation of the nuclear chromatin which takes place within the cell's physical compartment.[8][11]
- Chromatin condensation
The activation of condensation initiates at the periphery of the nuclear membrane, forming a dense crescent ring-like structure.
This is connected with the nuclear envelope, and later throughout the process, it is followed by clumping & margination of the chromatin structures, towards the inner nuclear membrane.[9]
Throughout the intermediate stage of pyknosis, specific contents of the cell undergoes cytoskeletal differentiation:
- present of cytoplasmic shrinkage
- nuclear matrix & the lamina endures degradation – causes a loss in the structural integrity & allows the formation of small osmophilic granules to aggregate throughout the whole nucleus.
- Compression of the osmophilic granules causes pressuring against the chromatin structures, in which, it generates a stress response adjacent to other constituents within the cell and causes retention of the protein fibrillar core.[8][9][12]
- DNA fragmentation
Progressively, with abundant cytoplasm condensed within the cell, it forms Extensive blebbing from the cell's plasma membrane, as a result,
it leads to the final stages of pyknosis & the initiation of karyorrheix. Initiation of karyorrheix leads to:
- Degradation of the cell's chromosomal DNA, whereby, chromosomal digestion results first in
large (30-50 and 200-300kbp) DNA fragments and,
- Nucleus of the cell undergoing internucleosomal fragmentation of the DNA, in which, the enzyme endonuclease is responsible for cleaving off the double stranded DNA.
- Disintegration of the cell's nucleus, degrades into small granular spheres termed Apoptotic Bodies.
The nucleus are surrounded by a nuclear membrane, containing variability amount of the cell's constituent (such as condensed cytoplasm, cytosolic elements, parts of the nuclei & crowded intact cytoplasmic organelles.) [8][11][12]
|Structural shrinkage of the cell
|Cell Membrane permeability
|Blebbing from the surface of the cell's plasma membrane
|Vacuole in the cytoplasm filled with cellular remanant
|Cell disintegrates into apoptotic bodies
|Diluation of the cellular organelles
|Phagocytosis by macrophages
|Loss of membrane integrity
|Total cell lysis
- Phagocytosis
Apoptotic bodies are engulfed by phagocytosis of adjacent cells - conducted by professional phagocytes of macrophages & parenchymal cells.
One of the integral morphological aspect of apoptosis is in the phagocytosis process, whereby, the process
of uptake & degradation of the apoptotic bodies is rapid due to, it prevent leakage of contents from the apoptotic bodies,
as a result it dissembles the mechanical pathway of apoptosis from eliciting an inflammatory response in the host.
Once phagocytosed, the apoptotic bodies are degraded by lysosomal enzymes derived from the ingesting cells and
therefore eventually being reduced to unrecognisable residual bodies.[9][11][12][13]
The morphology of necrosis
Necrosis is termed by the series of changes that accompany cell death, whereby, it occurs as a result of unintentional traumatic injury,
whether thermal, chemical, or anoxic.
Necrotic cells are unable to maintain their membrane integrity & results in leakage of contents and thus leads to eliciting an inflammatory response in the host.[13][14]
Initially, the hallmark of necrosis is cellular swelling, in which, the cells fail in exchanging cellular fluid from the intracellular & extracellular regions, as a result, it cells cannot gain energy of ATP for mechanical function and leads to the physiological response of cellular swelling.
In-addition, the morphology of the cell membrane becomes permeable throughout the cellular death phase.
There is a change in the cellular content of the cell with the loss of density, the organelles may dilute and ribosomes dissociates from adjacent endoplasmic reticulum, and finally the lost of membrane integrity leads to rupture and lysis of the cellular content into the surrounding environment, causing an pathological inflammatory response.[9][11][13]
Schematic drawing illustrating the main events of the biochemical & physiological pathway of apoptosis.
--Daniel Lee 16:54, 3 May 2009 (EST)
Design model of apoptosis
Apoptosis from a Clinical Point of View
As we now know that apoptosis follows a series of tightly regulated steps, it's important to consider the implications of malfunctions and irregularities. Interruptions and disruptions to the system are caused by many different human pathologies, many of which are by an unknown mechanism. The need to understand the causal effect of these different abnormalities is essential in the potential treatment of these diseases. Diseases which target this system include; autoimmune diseases, neurodegenerative diseases, cancer and others.
Autoimmune Diseases:
- There are two ways in which this utilises the apoptotic pathway;
- Inhibiting apoptosis in the clearing of B or T cells resulting in tissue destruction
- Inhibiting apoptosis in the destruction of cells displaying autoantigens on their cell surface (Blumenthal et al., 2004)
Neurodegenerative diseases:
- An example with Alzheimer's disease
- It has been found that the amyloid plaques (APP) are a substrate for caspase 3 resulting in defective protein and therefore reducing the inhibition of apoptosis (Robertson et al., 1999)
- Also the cleavage of APP by caspases 6, 8 and 9 generates the peptide C31, a potent inducer of apoptosis (Chandra et al., 2000)
- These factors play a pivotal role in the over-active apoptosis and hence the increase neurodegeneration as seen in Alzheimer's disease
- The use of capsase inhibition is proving to be effective in the protection against some neurodegenerative diseases, as well as from stroke, myocardial infarction, hepatic injury, spinal cord injury and many others.
- An example with IDN-6556, a selective and irreversible inhibitor of specific caspases used in liver injury. It shows potential in the application for liver transplants, as it helps restore many of the livers enzyme levels (Schulze-Osthoff et al., 2005)
- Cancer is almost apoptosis resistant
- Roughly half of all cancers affect p53 function, which is usually pro-apoptotic (Fadeel et al., 2005)
- iASPP (inhibitory apoptosis-stimulating protein of p53) has been found to be up-regulated in various cancers such as breast carcinomas (Fadeel et al., 2005)
- suppression of iASPP has been effective in inducing p53-dependent apoptosis – possibly a new strategy for the treatment of tumours expressing the wild-type p53 (O'Neil et al., 2003)
- Survivin is a protein of the IAP family which is expressed in cancers of the breast, colon, lung, pancreas and prostate. The inactivation of such a protein greatly increase the chance of therapeutic intervention (Altieri, 2003)
- Haematopoietic malignancies such as multiple myeloma and leukaemia are associated with a high ratio of Bcl-2 – BAX. By decreasing the concentrations of Bcl-2 and increasing BAX, the susceptibility of apoptosis occurring is increased in the cancerous cell. (Bentley et al., 1996; Wright el at., 1996)
- Chemotherapeutic drugs induce apoptosis with the aim of affecting the malignant cells more than the normal cells, unfortunately, some cancer cells are chemoresistant and many healthy cells are destroyed. The combination of these drugs with G-CSF, an anti-apoptotic factor for neutrophils, greatly decreases the apoptotic affects on neutrophils and hence the possibility of infection accompanying the chemotherapy (Nikolaizik et al., 1999; Yousefi et al., 2004). This combination therapy is also used with fibroblast growth factor, which reduces the gastrointestinal side-effects, and p53 inhibitors, which reduce normal tissue damage (Fadeel et al., 2005).
With the increasing understanding of what goes wrong with the apoptotic pathway comes the improved clinical effectiveness of the therapeutic intervention and prevention strategies. The future direction for the treatment of these various conditions lies predominantly in the improvement in selective targeting. The need to adapt and alter treatments specifically for the individual by the use of varying combination therapies is a huge focus, as there is an essential need to target the affected areas, whilst minimising the damage to normal areas. There has been a massive advancement in the understanding of the diseases affecting this fundamental pathway over the last 50 years, and there is still a long but optimistic future.
1. Lauber K, Blumenthal SG, Waibel M, Wesselborg S. (2004) Clearance of apoptotic cells: getting rid of the corpses. Molecular Cell. 14: 277–87.
2. Gervais FG, Xu D, Robertson GS. (1999). Involvement of caspases in proteolytic cleavage of Alzheimer's amyloid-b precursor protein and amyloidogenic Ab peptide formation. Cell. 1999; 97: 395–406.
3. Lu DC, Rabizadeh S, Chandra S. (2000) A second cytotoxic proteolytic peptide derived from amyloid b-protein precursor. Nat Med. 6: 397–404.
4. Bergamaschi D, Samuels Y, O'Neil NJ. (2003) iASPP oncoprotein is a key inhibitor of p53 conserved from worm to human. Nat Genet. 33: 162–7.
5. Altieri DC. (2003). Validating survivin as a cancer therapeutic target. Nat Rev Cancer 2003; 3: 46–54.
6. Pepper C, Bentley P, Hoy T. (1996). Regulation of clinical chemoresistance by bcl-2 and bax oncoproteins in B-cell chronic lymphocytic leukaemia. British Journal Haematology. 95: 513–7.
7. McConkey DJ, Chandra J, Wright S. (1996). Apoptosis sensitivity in chronic lymphocytic leukemia is determined by endogenous endonuclease content and relative expression of BCL-2 and BAX. Journal of Immunology. 156: 2624–30.
8. Dibbert B, Weber M, Nikolaizik WH. (1999). Cytokine-mediated Bax deficiency and consequent delayed neutrophil apoptosis: a general mechanism to accumulate effector cells in inflammation. Proc Natl Acad Sci USA. 96: 13330–5.
9. Altznauer F, Martinelli S, Yousefi S. (2004). Inflammationassociated cell cycle-independent block of apoptosis by surviving in terminally differentiated neutrophils. J Exp Med. 199: 1343–54.
10. Fischer U, Schulze-Osthoff K. (2005). Apoptosis-based therapies and drug targets. Cell Death Differ. 12: 942–61.
- ↑ 1.0 1.1 1.2 1.3 Reed JC. Dysregulation of apoptosis in cancer.J Clin Oncol. 1999 Sep;17(9):2941-53. Review. [1]
- ↑ Gross. A, McDonnell JM, Korsmeyer SJ.BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 1999 Aug 1;13(15):1899-911. [2]
- ↑ 3.0 3.1 Zinkel S, Gross A, Yang E. BCL2 family in DNA damage and cell cycle control. Cell Death Differ. 2006 Aug;13(8):1351-9. Epub 2006 Jun 9. [3]
- ↑ 4.0 4.1 Deveraux QL, Takahashi R, Salvesen GS, Reed JC.X-linked IAP is a direct inhibitor of cell-death proteases. GNature. 1997 Jul 17;388(6639):300-4. [4]
- ↑ Meike B. Pascal M. Ubiquitin-mediated regulation of apoptosis. 2009 Trends Cell Biol. Mar;19(3):130-40. [5]
- ↑ 6.0 6.1 Qin H, Yu T,Q et al. Regulation of apoptosis and differentiation by p53 in human embryonic stem cells.J Biol Chem. 2007 Feb 23;282(8):5842-52. [6]
- ↑ 7.0 7.1 Wyllie, A.H. (1987). APOPTOSIS: CELL DEATH IN TISSUE REGULATION. Journal of Pathology. 153(4), 313-316.
- ↑ 8.0 8.1 8.2 8.3 Doonan, F., & Cotter, T.G. (2008). Morphological assessment of apoptosis. Methods. 44(3), 200-204.
- ↑ 9.0 9.1 9.2 9.3 9.4 Saini, K.S., & Walker, N.I. (1998). Biochemical and molecular mechanisms regulating apoptosis. Molecular and cellular biochemistry. 178(1-2), 9-25.
- ↑ Bjelakovic, G., Nagorni, A., Bjelakovic, M., Stamenkovic, I., Arsic, R., & Katic, V. (2005). APOPTOSIS: PROGRAMMED CELL DEATH AND ITS CLINICAL IMPLICATIONS. Medicine and Biology. 12(1), 6-11.
- ↑ 11.0 11.1 11.2 11.3 Ziegler, U., & Groscurth, P. (2004). Morphological Features of Cell Death. News in physiological sciences : an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society. 19, 124-128.
- ↑ 12.0 12.1 12.2 Häcker, G. (2000). The morphology of apoptosis. Cell and tissue research. 301(1), 5-17.
- ↑ 13.0 13.1 13.2 Kumar, V., Abbas, A.K., Fausto, N., & Mitchell, R.N. (2007). ROBBINS Basic Pathology. (8th ed.). Philadelphia, United States of America: SAUNDERS ELSEVIER.
- ↑ McHugh, P., & Turina, M. (2006). Apoptosis and necrosis: a review for surgeons. Surgical infections. 7(1), 53-68.
- Trimerization (Term meanings still needed to be added)
- Caspase
- Lamins
- Gelsolin
- Fodrin
- Apoptosome
- Cytochrome c
2009 Group Projects
--Mark Hill 14:02, 19 March 2009 (EST) Please leave these links to all group projects at the bottom of your project page.
Group 1 Meiosis | Group 2 Cell Death - Apoptosis | Group 3 Cell Division | Group 4 Trk Receptors | Group 5 The Cell Cycle | Group 6 Golgi Apparatus | Group 7 Mitochondria | Group 8 Cell Death - Necrosis | Group 9 Nucleus | Group 10 Cell Shape


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Thomas L. Friedman's No. 1 bestseller The World Is Flat has helped millions of readers to see globalization in a new way. Now Friedman brings a fresh outlook to the crises of destabilizing climate change and rising competition for energy-both of which could poison our world if we do not act quickly and collectively. His argument speaks to all of us who are concerned about the state of America in the global future. Friedman proposes that an ambitious national strategy- which he calls "Geo-Greenism"-is not only what we need to save the planet from overheating; it is what we need to make America healthier, richer, more innovative, more productive, and more secure. As in The World Is Flat, he explains a new era-the Energy-Climate era-through an illuminating account of recent events. He shows how 9/11, Hurricane Katrina, and the flattening of the world by the Internet (which brought 3 billion new consumers onto the world stage) have combined to bring climate and energy issues to Main Street. But they have not gone very far down Main Street; the much-touted "green revolution" has hardly begun. With all that in mind, Friedman sets out the clean-technology breakthroughs we, and the world, will need; he shows that the ET (Energy Technology) revolution will be both transformative and disruptive; and he explains why America must lead this revolution-with the first Green President and a Green New Deal, spurred by the Greenest Generation. Hot, Flat, and Crowded is classic Thomas L. Friedman-fearless, incisive, forward-looking, and rich in surprising common sense about the world we live in today. Book Description
Thomas L. Friedmans phenomenal number-one bestseller The World Is Flat has helped millions of readers to see the world in a new way. In his brilliant, essential new book, Friedman takes a fresh and provocative look at two of the biggest challenges we face today: Americas surprising loss of focus and national purpose since 9/11; and the global environmental crisis, which is affecting everything from food to fuel to forests. In this groundbreaking account of where we stand now, he shows us how the solutions to these two big problems are linked--how we can restore the world and revive America at the same time.
Friedman explains how global warming, rapidly growing populations, and the astonishing expansion of the worlds middle class through globalization have produced a planet that is hot, flat, and crowded. Already the earth is being affected in ways that threaten to make it dangerously unstable. In just a few years, it will be too late to fix things--unless the United States steps up now and takes the lead in a worldwide effort to replace our wasteful, inefficient energy practices with a strategy for clean energy, energy efficiency, and conservation that Friedman calls Code Green.
This is a great challenge, Friedman explains, but also a great opportunity, and one that America cannot afford to miss. Not only is American leadership the key to the healing of the earth; it is also our best strategy for the renewal of America.
In vivid, entertaining chapters, Friedman makes it clear that the green revolution we need is like no revolution the world has seen. It will be the biggest innovation project in American history; it will be hard, not easy; and it will change everything from what you put into your car to what you see on your electric bill. But the payoff for America will be more than just cleaner air. It will inspire Americans to something we havent seen in a long time--nation-building in America--by summoning the intelligence, creativity, boldness, and concern for the common good that are our nations greatest natural resources.
Hot, Flat, and Crowded is classic Thomas L. Friedman: fearless, incisive, forward-looking, and rich in surprising common sense about the challenge--and the promise--of the future.
Thomas Friedman and Fareed Zakaria: Author One-to-One
Fareed Zakaria: Your book is about two things, the climate crisis and also about an American crisis. Why do you link the two?
Thomas Friedman: You're absolutely right--it is about two things. The book says, America has a problem and the world has a problem. The world's problem is that it's getting hot, flat and crowded and that convergence--that perfect storm--is driving a lot of negative trends. America's problem is that we've lost our way--we've lost our groove as a country. And the basic argument of the book is that we can solve our problem by taking the lead in solving the world's problem.
Zakaria: Explain what you mean by "hot, flat and crowded."
Friedman: There is a convergence of basically three large forces: one is global warming, which has been going on at a very slow pace since the industrial revolution; the second--what I call the flattening of the world--is a metaphor for the rise of middle-class citizens, from China to India to Brazil to Russia to Eastern Europe, who are beginning to consume like Americans. That's a blessing in so many ways--it's a blessing for global stability and for global growth. But it has enormous resource complications, if all these people--whom you've written about in your book, The Post American World--begin to consume like Americans. And lastly, global population growth simply refers to the steady growth of population in general, but at the same time the growth of more and more people able to live this middle-class lifestyle. Between now and 2020, the world's going to add another billion people. And their resource demands--at every level--are going to be enormous. I tell the story in the book how, if we give each one of the next billion people on the planet just one sixty-watt incandescent light bulb, what it will mean: the answer is that it will require about 20 new 500-megawatt coal-burning power plants. That's so they can each turn on just one light bulb!
Zakaria: In my book I talk about the "rise of the rest" and about the reality of how this rise of new powerful economic nations is completely changing the way the world works. Most everyone's efforts have been devoted to Kyoto-like solutions, with the idea of getting western countries to reduce their carbon dioxide emissions. But I grew to realize that the West was a sideshow. India and China will build hundreds of coal-fire power plants in the next ten years and the combined carbon dioxide emissions of those new plants alone are five times larger than the savings mandated by the Kyoto accords. What do you do with the Indias and Chinas of the world?
Friedman: I think there are two approaches. There has to be more understanding of the basic unfairness they feel. They feel like we sat down, had the hors d'oeuvres, ate the entre, pretty much finished off the dessert, invited them for tea and coffee and then said, "Let's split the bill." So I understand the big sense of unfairness--they feel that now that they have a chance to grow and reach with large numbers a whole new standard of living, we're basically telling them, "Your growth, and all the emissions it would add, is threatening the world's climate." At the same time, what I say to them--what I said to young Chinese most recently when I was just in China is this: Every time I come to China, young Chinese say to me, "Mr. Friedman, your country grew dirty for 150 years. Now it's our turn." And I say to them, "Yes, you're absolutely right, it's your turn. Grow as dirty as you want. Take your time. Because I think we probably just need about five years to invent all the new clean power technologies you're going to need as you choke to death, and we're going to come and sell them to you. And we're going to clean your clock in the next great global industry. So please, take your time. If you want to give us a five-year lead in the next great global industry, I will take five. If you want to give us ten, that would be even better. In other words, I know this is unfair, but I am here to tell you that in a world that's hot, flat and crowded, ET--energy technology--is going to be as big an industry as IT--information technology. Maybe even bigger. And who claims that industry--whose country and whose companies dominate that industry--I think is going to enjoy more national security, more economic security, more economic growth, a healthier population, and greater global respect, for that matter, as well. So you can sit back and say, it's not fair that we have to compete in this new industry, that we should get to grow dirty for a while, or you can do what you did in telecommunications, and that is try to leap-frog us. And that's really what I'm saying to them: this is a great economic opportunity. The game is still open. I want my country to win it--I'm not sure it will.
Zakaria: I'm struck by the point you make about energy technology. In my book I'm pretty optimistic about the United States. But the one area where I'm worried is actually ET. We do fantastically in biotech, we're doing fantastically in nanotechnology. But none of these new technologies have the kind of system-wide effect that information technology did. Energy does. If you want to find the next technological revolution you need to find an industry that transforms everything you do. B

subset: commoncrawl


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Akrylfärg Navy Cool Grey, 22 ml
22 ml färgburk med skruvkork.
All Billing Boats paints are Acrylic (Water based) Paints and are delivered in 22ml plastic bottles. We have spent 7 years in developing these paints making them authentic maritime colours. All numbers refer to the paints needed for any Billing Boats kit.


text:
The iWork Suite on the iOS has now finally found a home on the iPhone and iPod Touch, thanks to the recent universal update. Already considered to be among the most polished productivity apps on the platform, iWork users will no longer have to rely on their iPads when creating or editing documents while on-the-go. Aside from the universal support, the apps also include improved document management, allowing users to easily sort and organize docs into folders. Pages, Numbers and Keynote are sold separately for $9.99, but as always, if you already own these apps, the update can be downloaded for free. Full press release after the break.
Apple, Keynote – $9.99
Apple, Pages – $9.99
Apple, Numbers, – $9.99


text:
Police in Arlington, Texas, have arrested the mother of a toddler beaten to death after her body was found buried in Parker County.
Alexis Botello was arrested on two charges Tuesday, according to Arlington police.
The 17-year-old mother of Tylea Moore is charged with tampering with or fabricating physical evidence and injury to a child. Her bond is set at a total of $125,000.
Police believe Botello intentionally concealed Tylea's corpse, according to the search warrant affidavit.
Botello's boyfriend, Joshua Beard, 20, has been charged with capital murder in connection with the child's death.
Beard's aunt had originally called police to report the toddler missing. As officers continued to investigate, though, they suspected foul play.
U.S. & World
Police believe Beard beat the toddler inside the house he shared with Botello in the 2200 block of Polo Club Court.
Arlington police say Beard is accused of intentionally or knowingly causing Tylea's death "by hitting her with closed fists, open hands and stomping on her with his feet."
The warrant affidavit says that Botello told officers her boyfriend, Beard, killed her daughter. According to the search warrant, the couple drove to Springtown in Parker County and "found a bridge in a remote area. They dug a grave and buried Tylea."
Botello led investigators to the area where detectives found remains believed to belong to Tylea.
The remains have yet to be positively identified by the Tarrant County Medical Examiners Office.
It's not immediately clear if Botello or Beard has an attorney.
NBC 5's Julie Fine and Ray Villeda contributed to this report.


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Wednesday, September 30, 2009
Wordful Wednesday - Katy Bear
Every day at school my preschool daughter picks out a library book from the IMC to bring home. On Monday she picked out Curious George Goes to the Hospital. I thought it a rather odd choice but read it to her anyway on Monday night.
The next morning my husband woke up and went to feed and care for our cats. He noticed that Katy Bear looked sick and put some food in her bowl. She tried to walk, but couldn't. After consulting with the vet on the phone, we learned that her symptoms were consistent with a stroke and she probably wouldn't live. Before taking her into the veterinary office we talked with our daughter and told her that Katy was very sick and that she might not survive. With heavy hearts we had time to say some quick goodbyes. The vet put Katy to sleep.
As soon as my husband left for the vet's office, my daughter went to retrieve the Curious George book. She really wanted to understand what was happening to our cat, and I couldn't help but wish that Katy Bear would return just as Curious George had from the hospital. Sometimes books are good at explaining difficult topics but I don't own any that help explain the loss of a beloved pet. My husband and I adopted Katy 10 years ago, when she was just a kitten. We'll miss our mouse catcher. We'll miss our apple peel eating cat. We'll miss her shoe loving ways. We'll miss her.
Find more of this week's Wordless Wednesday (or Wordful) posts at 5 Minutes for Mom or Seven Clown Circus.
33 comments:
- C. Pinheiro said...
I'm so sorry to hear about your pet. Losing a pet is always hard, and after having one for a decade--well, it becomes so much harder.
It sounds like the Curious George book was a little touch of fate.
- September 29, 2009 at 11:55 PM
- KarieK said...
Oh I am so sorry. Gosh I should have brought tissue!
Sounds like Curious George was a great cure to explain such a difficult time and tragedy.
- September 30, 2009 at 1:20 AM
- Liz Mays said...
That's so sad. I'm sorry! I'm glad you had the book to help explain things.
- September 30, 2009 at 6:03 AM
- Jennifer said...
I am so sorry. (hug) Losing a pet is so difficult as they are one of the family.
- September 30, 2009 at 7:27 AM
- Wendy said...
I'm so sorry for your loss. Sounds like Curious George wasn't such a bad pick after all.
- September 30, 2009 at 8:56 AM
- HaB said...
Oh my goodness......Just last month, we put my beloved calico to sleep. It was heart wrenching - and so very hard to make our 2 year old daughter understand. My husband had given her to me the first Christmas we were together as boy/girl friend almost 10 years ago. Very similar story to your own.
BIG HUGS to you for the loss of what clearly was a very well loved and treasured member of you family.
- September 30, 2009 at 9:25 AM
- Susie said...
What a sad but teachable moment!
- September 30, 2009 at 10:01 AM
- Michelle said...
I'm so sorry you had to put your kitty to sleep. I had to do the same thing with our first cat, but my oldest was too young at the time to need to explain anything to her. Good thing you had George to help you out.
- September 30, 2009 at 10:07 AM
- tiarastantrums said...
aww - so sorry for your loss!
- September 30, 2009 at 10:14 AM
- Shaggy and Scout said...
Your blog title caught my eye over at Daisy's blog, so I thought I'd come visit.
What sad news to find! I'm am so sorry you had to say goodbye to your beloved Katy. Please know that she will forever be in your hearts.
sss's mom
- September 30, 2009 at 10:16 AM
- Melanie @ Whimsical Creations said...
I am so sorry for your loss. Our kitty had a stroke last November and passed away. Explaining to children is awful. HUgs!!
- September 30, 2009 at 10:20 AM
- Samantha & Mom said...
Hugs and purrrs to you all!! Our thoughts are with you!! W are glad George was there to help!!
Your TX furiends,
- September 30, 2009 at 10:34 AM
- Daisy said...
I am very sorry that Katy Bear had to go to the Rainbow Bridge. She was very beautiful.
- September 30, 2009 at 10:37 AM
- Sherry said...
sorry to hear of your loss! :( best regards!
- September 30, 2009 at 10:40 AM
- Cat with a Garden said...
We are very sorry to hear that Katy Bear had to be put to sleep. She didn't suffer for long though and that's a comfort we think. It must be very hard for all of you. Your little girl was lucky she could say her goodbyes to Katy Bear.
Gentle purrs, Siena & Chilli
- September 30, 2009 at 11:11 AM
- Jennifer-Eighty MPH Mom said...
I am so, so sorry. I am in tears now. Katy Bear sounds like a special girl (beautiful too). I am a kitty person myself, and I know how much this hurts. They really are our second children and they are loved so much.
I am so sorry.
- September 30, 2009 at 11:19 AM
- Andrea said...
I'm so sorry for your loss! Pets are truly part of the family, and it's truly a shame they don't live as long as we do! I don't know what I'll do when we eventually lose one of our pets!
- September 30, 2009 at 11:57 AM
- Thoughts said...
Oh no, I am SO SORRY to hear about your loss. We recently lost our 19-year-old housecat so I know what you are going through.
I am sending warm thoughts and prayers your way and so are all the furbabies in my family...
- September 30, 2009 at 12:06 PM
- Annie Bear said...
I'm so sorry about Katy. She's so pretty and sweet looking.
Thanks for stopping by my blog.
- September 30, 2009 at 12:27 PM
- Tara said...
ohhhh so sorry to hear about your darling cat!! what a sweet little story though...
- September 30, 2009 at 2:02 PM
- TheAtticGirl said...
So sorry to hear about your cat! It's hard when you lose a member of the family. Praying for you!
- September 30, 2009 at 3:19 PM
- Jenny said...
I have been in your shoes and can appreciate your pain. I remember telling the kids was the hardest part. Blessings to you.
- September 30, 2009 at 3:50 PM
- Brenda said...
How very sad, I am so sorry to hear about your loss.
- September 30, 2009 at 4:35 PM
- Alesha @ Full Time Mama said...
Oh, I am SO sorry! She was a gorgeous cat! It's so hard losing a pet, especially one you've had for as long as you had her. Hugs!
- September 30, 2009 at 6:28 PM
- Kristin said...
I'm sorry!
- September 30, 2009 at 7:21 PM
- Staci A said...
I'm so sorry to hear. I can't imagine how tough it must be!
- September 30, 2009 at 7:56 PM
- Connie said...
Our furry family members provide us with so much love, it's hard to say good-bye. Hugs to you all as you remember your sweet kitty.
- September 30, 2009 at 8:02 PM
- Carol M said...
I have tears in my eyes. I'm so sorry for the loss of your kitty. I've gone through it three times now and I know how much it hurts.
Carol M
- September 30, 2009 at 8:04 PM
- Mikki said...
Aww. Poor thing. It is so hard to loose a family pet. That's what they become.. a part of the family.
Praying that you all can laugh at all the good memories of your mouse catcher :-)
- September 30, 2009 at 8:33 PM
- Adaptable Kay said...
Aw, I am so sorry! It hurts all the same whether it's an animal or a person.
At least you have your memories =]
- September 30, 2009 at 9:24 PM
- Muthering Heights said...
That is so sad! I'm so sorry. :(
- September 30, 2009 at 10:12 PM
- Marie said...
I'm so sorry for your loss. It's terrible to lose an animal who has become part of your family -- I still miss our Tuckers who we lost a few years ago now. The vet said it was either a stroke or a brain tumor :-(
One book that my children really enjoy is called Kitty Heaven. I got it for them when we lost Tuckers and they still ask to read it periodically so they can see that Tuckers is doing well :-) It's a very sweet book.
- October 1, 2009 at 7:10 PM
- Eli's Lids said...
I still miss my first kitty, Ringo!! What a blessing your daughter picked out that book... hopefully it helped a little!
- October 4, 2009 at 10:32 AM


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Casa Bacardi Visitor Center
Visit the slick Casa Bacardi Visitor Center (t787/788-8400, wwww.casabacardi.org) inside the "cathedral of rum", the vast Bacardi distillery across San Juan Bay in Cataño and you'll enter another world – Cuba, to be precise. It's a series of fun and illuminating interactive exhibits that emphasize Bacardi's Cuban roots and involve not just watching and listening, but sniffing the products on display. Guided tours depart every 15 to 30 minutes and last around 45 minutes.
Established in Santiago de Cuba by Catalan expat Don Facundo Bacardí Massó in 1862, the Bacardi empire now dominates the global rum market, supplying 75 percent of rum sold in the US alone. The Puerto Rican plant was established in Old San Juan in 1936 and moved to this location – when it received its "cathedral" sobriquet from then-governor Luis Muñoz Marín – in 1958. The move here proved timely, as Castro seized the Bacardi assets in Cuba shortly afterwards, precipitating exile in 1960 – the family remain vehement opponents of what they term a "totalitarian" regime.
Today Bacardi is a true multinational organization, headquartered in tax-free Bermuda, and with massive operations in the Bahamas, Mexico and Puerto Rico – the last outpost has the capacity to produce 100,000 gallons of rum every 24 hours and is the biggest taxpayer on the island.
Equipped with a hand-held audio guide and accompanied by enthusiastic docents, you'll pass through seven different zones introducing both the history of the company and the rum-making process. Special barrels allow you to "nose" the effects of wood barrelling, ageing and finishing, as well as the various Bacardi brands on offer: sweetly scented apple and melon flavours and the rich, addictive aroma of coconut-laced rum – piña colada in a bottle. Mercifully, there are two free drinks waiting for you at the end of the tour (and as many soft drinks as you like).
Note, however, that you don't get to visit the actual distillery – for security reasons the real rum-making facilities have been off-limits since 9/11 and are likely to remain that way.
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Welcome to our health education library. The information shared below is provided to you as an educational and informational source only and is not intended to replace a medical examination or consultation, or medical advice given to you by a physician or medical professional.
Follow your doctor's advice about caring for your hemorrhoids at home. Some treatments help relieve symptoms right away. Others involve making changes in your diet and exercise habits. These can help ease constipation and prevent hemorrhoid symptoms from coming back.
Your doctor may prescribe anti-inflammatory medication to help ease your symptoms. The following tips will also help relieve pain and swelling.
Take Sitz Baths. Taking a sitz bath means sitting in a few inches of warm bath water. Soaking for 15-20 minutes twice a day can provide welcome relief from painful hemorrhoids. It can also help the area stay clean.
Develop Good Bowel Habits. Use the bathroom when you need to. Don't ignore the urge to go. This can lead to constipation, hard stools, and straining. Also, don't read while on the toilet. Sit only as long as needed. Wipe gently with soft, unscented toilet tissue or baby wipes.
Use Ice Packs. Placing an ice pack on a thrombosed external hemorrhoid can help relieve pain right away. It will also help reduce the blood clot. Use the ice for 15-20 minutes at a time. Keep a cloth between the ice and your skin to prevent skin damage.
Use Other Measures. Laxatives and enemas can help ease constipation. But use them only on your doctor's advice. For symptom relief, try using cotton pads soaked in witch hazel. These are available at most drugstores. Over-the-counter hemorrhoid ointments and petroleum jelly can also provide relief.
Adding fiber to your diet can help relieve constipation by making stools softer and easier to pass. To increase your fiber intake, your doctor may recommend a bulking agent, such as psyllium. This is a high-fiber supplement available at most grocery and drugstores. Eating more fiber-rich foods will also help. There are two types of fiber:
Insoluble fiber is the main ingredient in bulking agents. It's also found in foods such as wheat bran, whole-grain breads, fresh fruits, and vegetables.
Soluble fiber is found in foods such as oat bran. Although soluble fiber is good for you, it may not ease constipation as much as foods high in insoluble fiber.
Along with a high-fiber diet, drinking more water can help ease constipation. This is because insoluble fiber absorbs water, making stools soft and bulky. Be sure to drink plenty of water throughout the day. Drinking fruit juices, such as prune juice or apple juice, can also help prevent constipation.
Regular exercise aids digestion and helps prevent constipation. It's also great for your health. So talk with your doctor about starting an exercise program. Low-impact activities, such as swimming or walking, are good places to start. Take it easy at first. And remember to drink plenty of water when you exercise.
High-fiber foods offer many benefits. By making your stools softer, they help heal and prevent swollen hemorrhoids. They may also help reduce the risk of colon and rectal cancer. Best of all, they're usually low in calories and taste great. Here are some examples of fiber-rich foods.
Whole grains, such as wheat bran, corn bran, and brown rice.
Vegetables, especially carrots, broccoli, cabbage, and peas.
Fruits, such as apples, bananas, raisins, peaches, and pears.
Nuts and legumes, especially peanuts, lentils, and kidney beans.
The tips below offer some simple ways to add more high-fiber foods to your meals.
Start your day with a high-fiber breakfast. Eat a wheat bran cereal along with a sliced banana. Or, try peanut butter on whole-wheat toast.
Eat carrot sticks for snacks. They're easy to prepare, taste great, and are low in calories.
Use whole-grain breads instead of white bread for sandwiches.
Eat fruits for treats. Try an apple and some raisins instead of a candy bar.
Discover leading-edge gastroenterology care. Call Digestive Disease Consultants of Orange County at 949.612.9090 or simply use the Request an Appointment form.


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Didj Make ATAP
creating an atap cartridge on an NHE board.
Warning, flashing your didj or the cart could brick it, you have been warned
If you have a modified game cart with the otp chip replaced with a rw nand chip or a NHE cart then this tutorial is for you, this is by no means complete but should get you well on your way to creating your own recovery cartridge. It will also show you the beginnings of being able to create our own partition structure for the nand devices, which in turn should eventually lead us to being able to run a full lx kernel and rootfs without having to do too much to the codebase.
Some form of writable nand cartridge Didj-Linux-4222-20090422-1236 sources ftdi cable bootstrap-LF_LF1000-0.33.5.3186_ATAP.lfp firmware-LF_LF1000-0.33.5.3186-ATAP.lfp .lfp files are in /Didj-Linux-4222-20090422-1236/packages/mfg-cart/Base2ATAP/Atap_FW
Preparing your didj to flash the cart
we need to get a custom firmware onto the Didj so that it can 'see' the extra mtd partitions on the cart, fortunately for us, leapfrog provided those in the form of the bootstrap/firmware ATAP.lfp files in /Didj-Linux-4222-20090422-1236/packages/mfg-cart/Base2ATAP/Atap_FW :)
mount the /Didj partition on your ubuntu system:
dmesg|tail sudo ./scsi_custom -c unlock /dev/sg<sg number returned from dmesg, usually sg2>
if you don't want to do the scsi_custom command you can do the following:
echo "UNLOCKED" > /flags/usb_mass_storage
bear in mind that this will leave your didj in a permanent state of readiness, as soon as you plug your didj cable in it should automount, although it doesn't auto-remount when you reboot, simply unplug/replug the cable and your devices will show up in ubuntu.
unpack each of the lfp files into /Base/
you'll end up with 2 extra folders in /Base:
/Didj/Base/bootstrap-LF_LF1000/ /Didj/Base/firmware-LF_LF1000/
bootstrap contains a lightning boot, firmware contains the kernel and rootfs files
make sure you are booted from RFS0 (we'll keep this as the original for now) and make sure the didj knows that you've got a mfcart in the unit, once done reboot:
echo RFS0 > /flags/rootfs touch /flags/mfcart reboot
Flashing the didj with the atap files will load a bootloader (not lb, but no reason you can't make it do it?) kernel1 and RFS1 to your didj's nand.
I'm not certain that you need to update the bootloader but for the sake of completeness I did, on the didj do the following to update the bootloader and firmware:
blupdate fwupdate
reboot and you should now be in the ATAP firmware on RFS1, you should see an extra 9 mtd partitions being created.
NAND device: Manufacturer ID: 0x2c, Chip ID: 0xda (ST Micro NAND 256MiB 3,3V 8-bit) Scanning device for bad blocks lf1000-nand: cartridge ECC mode: <6>software NAND device: Manufacturer ID: 0x20, Chip ID: 0xdc (ST Micro NAND 512MiB 3,3V 8-bit) Scanning device for bad blocks 9 cmdlinepart partitions found on MTD device lf1000-cart Creating 9 MTD partitions on "lf1000-base": 0x00000000-0x00020000 : "LF1000_uniboot" 0x00020000-0x00100000 : "Atomic_Boot_Flags" 0x00100000-0x00200000 : "Manufacturing_Data" 0x00200000-0x00400000 : "Kernel0" 0x00400000-0x01200000 : "Linux_RFS0" 0x01200000-0x01400000 : "Kernel1" 0x01400000-0x02200000 : "Linux_RFS1" 0x02200000-0x10000000 : "Brio" 0x10000000-0x10000000 : "EXT" mtd: partition "EXT" is out of reach -- disabled Creating 9 MTD partitions on "lf1000-cart": 0x00000000-0x00020000 : "prg_LF1000_uniboot" 0x00020000-0x00100000 : "prg_Atomic_Boot_Flags" 0x00100000-0x00200000 : "prg_Manufacturing_Data" 0x00200000-0x00400000 : "prg_Kernel0" 0x00400000-0x01200000 : "prg_Linux_RFS0" 0x01200000-0x01400000 : "prg_Kernel1" 0x01400000-0x02200000 : "prg_Linux_RFS1" 0x02200000-0x10000000 : "prg_Brio" 0x10000000-0x20000000 : "prg_EXT"
doing a cat /proc/mtd shows us that the mappings are not direct 1:1, our Brio/Didj partition gets moved all the way to mtd18 for some reason, not sure why it does this but I think it creates an ubi volume it then creates an extra mtd partition and mounts either the ubi volume to it or the new mtd partition to the ubi volume. It does more of the same to the cart once its formatted correctly. More on this later
# cat /proc/mtd dev: size erasesize name mtd0: 00020000 00020000 "LF1000_uniboot" mtd1: 000e0000 00020000 "Atomic_Boot_Flags" mtd2: 00100000 00020000 "Manufacturing_Data" mtd3: 00200000 00020000 "Kernel0" mtd4: 00e00000 00020000 "Linux_RFS0" mtd5: 00200000 00020000 "Kernel1" mtd6: 00e00000 00020000 "Linux_RFS1" mtd7: 0de00000 00020000 "Brio" mtd8: 00000000 00020000 "EXT" mtd9: 00020000 00020000 "prg_LF1000_uniboot" mtd10: 000e0000 00020000 "prg_Atomic_Boot_Flags" mtd11: 00100000 00020000 "prg_Manufacturing_Data" mtd12: 00200000 00020000 "prg_Kernel0" mtd13: 00e00000 00020000 "prg_Linux_RFS0" mtd14: 00200000 00020000 "prg_Kernel1" mtd15: 00e00000 00020000 "prg_Linux_RFS1" mtd16: 0de00000 00020000 "prg_Brio" mtd17: 10000000 00020000 "prg_EXT" mtd18: 0d812000 0001f800 "ubi_Brio"
Preparing the mtd partitions for mkbase.sh
mkbase.sh will flash the nhe cart with the same nand layout as the didj nand, at this moment in time the partitions don't exist, so it will fail, the flash_eraseall functions don't seem to work correctly so we do it ourselves.
We need to erase the following partitions:
mtd13: 00e00000 00020000 "prg_Linux_RFS0" mtd15: 00e00000 00020000 "prg_Linux_RFS1" mtd16: 0de00000 00020000 "prg_Brio" mtd17: 10000000 00020000 "prg_EXT"
I'll show you the first one (prg_Linux_RFS0) as an example, to do this we use mtd_debug on the didj to get mtd.size:
# mtd_debug info /dev/mtd13 mtd.type = MTD_NANDFLASH mtd.flags = MTD_CAP_NANDFLASH mtd.size = 14680064 (14M) mtd.erasesize = 131072 (128K) mtd.writesize = 2048 (2K) mtd.oobsize = 64 regions = 0
then on your ubuntu machine convert the mtd.size to something we can use in mtd_debug:
# echo 'obase=16; 14680064' | bc E00000
then on the didj do:
mtd_debug erase /dev/mtd13 0 0x00E00000
repeat this whole process for mtd15,16 and 17, then reboot the didj, you will see it complain that it couldn't find something on prg_EXT, and it will format it for you (it might do the same to mtdblock16) if not then do:
mkfs.vfat -n Didj -S 512 /dev/mtdblock16
Run the mkbase.sh script
at this point you should be ready for mkbase.sh to run to completion. give it a try, from the didj do:
# mkbase.sh Mounting /dev/mtdblock18 on /Didj as rw Mounting /dev/mtdblock19 on /opt/prg_mfg as rw /dev/mtd9 is the bootstrap partition /dev/mtd12 and /dev/mtd14 are the kernel partitions /dev/mtd13 and /dev/mtd15 are the root partitions /dev/mtd11 is the manufacturing partition /dev/mtdblock11 is the manufacturing partition block device /dev/mtd10 is the flags partition /dev/mtdblock10 is the flags partition block device /dev/mtd16 is the brio partition /dev/mtdblock2 11 is the base mfg partition /dev/mtd17 is the extended partition /dev/mtdblock17 is the extended partition mount device 10000000 is the extended partition size I have found a development cartridge Making a manufacturing cartridge BOOTLDR=/Didj/Base/bootstrap-LF_LF1000/lightning-boot.bin FLAGS=/Didj/Base/bootstrap-LF_LF1000/bootflags.jffs2 KERNEL=/Didj/Base/firmware-LF_LF1000/kernel.bin ROOTFS=/Didj/Base/firmware-LF_LF1000/erootfs.jffs2 Boot sector programmed successfully Flag partition programmed successfully Linux Kernel partition 0 programmed successfully Linux Kernel partition 1 programmed successfully Skipping manufacturing partition Root filesystem partition 0 programmed successfully Root filesystem partition 1 programmed successfully End of root filesystem setup Configuring your manufacturing cartridge... Empty flash at 0x0000007c ends at 0x00000800 No startup scripts. Cartridge will be a bare boot cartridge. No files found to preload the ATAP cartridge with. Programming complete
As you can see, there are a couple of issues, No Startup scripts and No files found to preload proably mean there are some more bits we need to to fill in to make this a completely working atap cartridge.
you should now do a reboot, check whether the cart prg_EXT partition has been mounted on the didj:
# mount rootfs on / type rootfs (rw) /dev/root on / type jffs2 (rw) none on /proc type proc (rw) sysfs on /sys type sysfs (rw) /dev/ram0 on /tmp type tmpfs (rw) /dev/mtdblock1 on /flags type jffs2 (rw,sync,noatime) /dev/mtdblock2 on /mfgdata type jffs2 (rw,sync,noatime) /dev/mtdblock18 on /Didj type vfat (rw,noatime,fmask=0022,dmask=0022,codepage=cp437,iocharset=i) /dev/mtdblock19 on /opt/prg_mfg type vfat (rw,noatime,fmask=0022,dmask=0022,codepage=cp437,ioch)
our prg_EXT partition was on mtd17, its now been mounted as an ubi volume to mtdblock19 you can find it in /opt/prg_mfg, this is similar behavour to the Brio:/Didj partition, it starts on mtd7, without a cart its put onto mtdblock9 as an ubi volume, with a cart /Didj ends up on mtdblock18!!
do a reboot, then plug your usb cable in and mount the partitions in ubuntu(unecessary if you did the 'echo UNLOCKED > /flags/usb_mass_storage' command earlier on':
dmesg|tail sudo ./scsi_custom -c unlock /dev/sg3
You'll notice that we get sdb and sdc which translated to sg2 and sg3, you should now see both the Brio:/Didj partition and the prg_EXT:/opt/prg_mfg partitions, /Didj will be 'DIDJ' and /opt/prg_mfg will be 'MFG_PAYLOAD'
I think we need to look at all of the /scripts files to glean more information about how to truly make an atap cart, I think make_release.sh shows us how to package some files for the mfg-cart ATAP stuff so they can be installed on the didj, I think we can probably make up the rest of the files needed from mfg-cart and leapfrog lfp files
What next and what have we found out?
It would obviously be nice to make the prg_Brio partition show up automatically, I'd like to work out how we can get it to show up as an ubi volume, not entirely sure what the mechanism is there yet.
We should be able to boot from the cart, not sure how that works yet either, but something along the lines of:
echo RFS2 > /flags/rootfs
should be enough, it will be in the sources, just need to track it down (lightning boot uses RFS0, RFS1, NFS0, NFS1 currently, will probably need extra flags)
I'd like to make a full cart, there are some bits missing, which I think are in the mfg-cart folders or at least the info.
I think we've learnt how to format a nhe cart, with the same partition structure as a didj nand unit, so we now know how to roughly deal with rebuilding a didj, so by extension we are not that far removed from being able to rebuild an LX partartition structure on a didj.
If you do cat /proc/cmdline you will notice that it builds the mtd partition structure for the cart in there, I think we should be able to use the same technique to make a custom SD booting kernel that can see a cart, this way we probably wouldn't need to flash the didj at all with the atap firmware. we can use root=31:04 technique to boot from the NHE, for instance from 31:12 :) once we know that we can boot from a cart we should be able to leverage an lx kernel with sufficient mtd2/ubi utils (will need to make ubiformat etc. ourselves from the new-utils).
We should probably look at a lot more of the scripts that are dotted all over the sources, the emeraldmfgtest folder found on an original lx firmware gives us some hints, as does make_release.sh, mkbase.sh mkSuperATAP.sh and many others.


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Saturday! I made the mistake of trying to cross the floor...it took about an hour. But...
I learned that Stephan Martiniere has a 2007 calendar out, Futurescapes. I'm proud to say that six of the twelve months are Tor book covers.
Brom told me that he is hard at work on a new novel. He was not at liberty to reveal the title but, like The Plucker, it will be lavishly illustrated.
Dan Dos Santos' painting demo.
Saturday, July 22, 2006
Comic Con: Saturday
Labels: Conventions, Stephan Martiniere
3 comments:
- mike said...
hey irene,
im really digging your blog! especially the up to the minute reporting of the comicon, and the helpful tips for breakin in to the industry.
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I know that many people have already given their, largely unfavorable, opinion on this asinine new "tool" that has entered the game, but I thought it was time for me to pitch in my two cents on the matter. For those of you that aren't interested in another gearscore rant, here is the TL:DR version: Gearscore can kiss my fuzzy tauren ass.
Over our holiday weekend, I partook in any number of PuGs through the looking for group system on just about all of my 5 level 80s. I've done a good number of these on Beru, so that I could finish off my achievement and nab up Perky (which I did this weekend!). Brade and I also always make a point to do the daily random for our two badges of frost. One of these days, our queue popped us into the Culling of Stratholme, which is one of my favorite instances.
As we entered the dungeon we discovered we were grouped with a Death Knight, Rogue and Shadow Priest. I often greet my groups with a bit of schtick, largely because I like to break the ice and I like to encourage communication in the group. I am usually fairly silly, and informative, going with something like "Hello, I will be your cruise director for this evening's adventures. We will be traveling at high speeds and will only slow down at passenger request. Please keep your seats in their full upright and locked positions, and keep your hands in the vehicle at all times". It is generally met with a laugh, and the groups get off on a good foot.
It also worked in our Culling of Stratholme group, but before we got underway Mr. Shadowpriest starts complaining about how we have two people in the raid with less than 2k "GS". Brade and I both reply with a "and?" response. The priest starts complaining about how he wants his extra badges, and he doesn't think the DPS can get there in time. Brade assures him that this will not be an issue (Brade has taken to tanking most of the heroics in full DPS gear, and often times tops damage as the tank) as he can't remember the last time he didn't make the timer.
At this point, the DK, who just hit 80 the other day, starts profusely apologizing about his gear, explaining he had just leveled and knew he didn't have the best gear. This made me furious. Not at the DK, but at the priest for being such a pretentious asshole and making the DK feel bad for ENTERING AN INSTANCE MEANT FOR HIM TO GEAR UP. I told the DK that he "better not dare apologize, I thought he would do just fine, and that these instances are meant to get him gear and he had every right to be here and I am happy to help him".
Of course, it didn't take long for the priest to check out the gear score of the mouthy druid in the group. This was met with a "Holy Shit, I don't think I've ever seen a GS this high before"…and all of the sudden I was "fabulous" just like that. I replied with "well, there you have it, I have the best gear score and I don't mind our DK, so you shouldn't either".
We finished the instance without event, having somethings obscene like 8 minutes left on the timer by the time we got to the end. Brade doubled the shadow priests damage, and you could almost add up everyone else's damage and Brade had still done more. But you know what? Who bloody well cares?! We finished the instance, and nobody did anything completely stupid. Even with our two sub 2k scored people.
The priest, somewhat humbled, politely thanked us for "carrying" them through the instance and went on his way. Of course, the DK and rogue were curious if we were going to keep running and wanted to tag along 🙂
It has been said a million times: Gear does not equal skill. Gearscore has got to be the most ass backwards thing that I have ever seen come out of this game. The person that developed the add-on should be hexed and made to live as a toad for the remainder of his time in WoW.
Of course, he isn't solely to blame. The members of the community have let it get to their head, and are quite out of control. Hell, one of our officers was telling me that he was in a group the other day with a DK in full epic healing plate. But…his gearcore ROCKED! He couldn't do shit all for damage, but man did his gearscore own! And I'm sorry…but you do NOT need full epics to run the pre-3.3 heroics! I'll even go one step further and say that you don't even need full epics for any of the new ones save Halls of Reflection, which admittedly needs a fairly well geared group. And by "well geared" I mean what you can get out of the preceeding dungeons, NOT full ICC/ToC gear.
In short, I wish this mod would die in a fire. You cannot evaluate how someone is as a player based solely on their gear. Period. It really makes me angry that other players make fresh 80s feel bad for running instances to gear up. It is completely ridiculous. Nobody should be made to feel guilty for running instances designed to be done in BLUES. I am actually tempted to start taking off a piece of gear each time someone mentions "GS" in one of my groups. GRRRRR. I really try to be very plesant in my PuGs, but as soon as someone starts pulling out gearscore shit, this tauren tends to become a mother bear: hostile and defensive of those that need protection from your stupidity.
In other things that irritated Beru during her PuG Adventures:
Brade was doing an ICC 10 on his paladin, so I was running the new ICC PuGs in my futile attempts to get a Hilt. (I've seen two drop, and lost the roll on both, but both to guild mates so I'm coping!). I ended up in a Heroic Halls of Reflection with my paladin. I actually really like healing this zone with the paladin because I just have so many tools available to save people's asses. Not to mention that beacon is god like. Need to heal the idiot DPS that isn't on the right target and taking tons of damage? No problem! The tank is never at risk of dying because you had to toss someone else a heal.
Sorry, I digress.
So, here I am healing along my merry way, and we finish the second boss making it through all of those god awful waves without a single death or wipe. And the tank, a druid, says to me in party chat: "You should only use Flash of Light, you are healing way too much". Ahem. What? I am sitting here thinking to myself…I have 5 years of healing experience and 4 healers. Nobody died and I didn't run out of mana. What in the fuck is this guy going on about? Not to mention that yes…there was need for using Holy Light. LIKE WHEN YOU ARE BELOW 50% OF YOUR LIFE. Oh yea…and did you ever stop to think that perhaps you are getting a Holy Light via beacon? BECAUSE ONE OF THE DPS PULLED THREAT FROM YOU AND IS ALMOST DEAD BECAUSE YOU DIDN"T NOTICE?
In the interest of not unleashing the inner bitch that is starting to rage inside of me, I kept my mouth shut. Mr. Druid, however, does not and spams his overhealing meter. Alright dickhead, it's on. "Hi, I'm a paladin. I overheal. It happens". I do not claim to be an expert on paladin healing, but I like to think that I am at least a competent paladin. I mean, ffs, I have two viable heals: One for 6k and one for 18k. I do not get an in-between, so either you don't get healed enough or you get healed too much. Guess which one I'd rather have happen?! He keeps going on about it. I am clearly not the only one annoyed as the shaman in the group pipes up "Overhealing is bad? Nobody died, who cares". The druid is still going on. So, I finally say, "Perhaps you would like to let me tank and you can heal and then I will criticize your healing for you?".
That shut him right up. I am still trying to decide if he just needed something to bitch about because the group ran fairly clean…and he needed to complain about SOMETHING. Or perhaps this was an alt and his main was a paladin healer and he thought "ur doin' it rong". I have no idea. As a general rule, it would seem to me that if someone appears to be doing their job properly, it is probably best NOT to give them shit about how they are getting the job done.
All of that bitching aside…
I still really do like the LFG tool. It is a massive improvement from what the game started with, and its many iterations since. I certainly have more good experiences than bad. It's just that the bad stick out more clearly in my memory 🙂
How about you? How have your experiences been? Have you had any strange experiences? Are you ready for "GS" to be "so 2009"?


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Rumor: Would 'T-Mac' fit with Celtics?
If he was willing to accept a bench role, the Celtics could find minutes to utilize his skill set and really allow him to thrive without the weight of having to be pre-2007 McGrady.
HoopsVibe's Very Quick Call: Would he accept a role?
Tracy McGrady heading to the Boston Celtics sounds good, however, there's the risk the former superstar won't accept less minutes, less shots, and less touches with Doc Rivers' squad.
After all, McGrady still, to some extent, believes he's 'T-Mac', the seven-time All-Star and two-time scoring champ. Last year McGrady, despite being past-his-prime, acted like a spoiled first-option and undermined Detroit Pistons coach John Kuester.
Such behaviour would never fly in Boston. He'd have to check his ego. He'd have to sit and watch his peers –veteran stars like Ray Allen, Kevin Garnett, and Paul Pierce – play. And he'd have to come off-the-bench.
In short, he'd have to become a professional. Nothing else would be tolerated by Rivers and GM Danny Ainge.
Of course, there is an alternative. McGrady can sign with another rebuilding squad and try parlaying the extra minutes and extra shots into a final, decent-money contract.
That would mean more meaningless basketball, though. In fourteen seasons, McGrady has never made the second round –ever. There was the choke-job with Orlando; there were tears in Houston.
Hopefully, McGrady has pride. Hopefully, he considers Boston. Hopefully, he becomes a professional –if he signs. It's not too late to salvage what he can.
–Oly Sandor.
Got thoughts? Well, get at HoopsVibe News in the comment box below.


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Full coverage of the Pistorius trial
Pretoria: Edging towards the end of his cross-examination, prosecutor Gerrie Nel asked Oscar Pistorius who the world should blame for the athlete's actions in shooting Reeva Steenkamp dead.
After all, the wily Mr Nel said, Mr Pistorius says he is blameless because he just made a "terrible mistake" that night, reacting instinctively to a noise like "wood moving" in his toilet cubicle that he thought was made by a dangerous intruder.
"So who should we blame for the fact that you shot her?" Mr Nel said, casting his arm around the court, as though anyone would do. "Should we blame Reeva? She never told you she was going to the toilet, should we not blame her?"
"No, my lady," came the quick response, addressed to the judge.
Was it the government then, Mr Nel continued, for not doing enough about South Africa's crime problem, leaving the disabled athlete apparently so paranoid of attack that he would shoot at a closed bathroom door?
"I don't know who [is to blame], my lady," he said.
"You must be blaming somebody for this?"
"I don't, my lady, I believed there was a threat."
Mr Pistorius' penchant for blame-shifting had been a recurring theme over the course of Mr Nel's tough cross-examination, and the questions were a fitting way to end his take-down of the international sporting icon.
After days in full attack mode, Mr Nel's climax was simply stated.
"You shot four shots though that door knowing she was talking to you … you armed yourself with the sole purpose of shooting and killing her," Mr Nel said.
"That is not true, my lady," Mr Pistorius replied.
"And after that you were overcome by what you had done … because you had intended to kill her?"
"That is not true, my lady."
For five days, it had been the kind of clash relished by sports lovers as much as court watchers – the Pit Bull prosecutor and the Blade Runner engaged in a battle that could have only one winner.
If the look on Mr Pistorius' face was anything to go by as he slumped back in the dock after seven days in the witness box, the answer was obvious.
Mr Nel had been his tormenter: the experienced prosecutor's precision cross-examination laid out a compelling argument that Mr Pistorius' version of events is utterly implausible.
Mr Pistorius appeared to be caught out and confused on numerous occasions, flatly denying minor facts that were easily proven.
Nonetheless he stuck to his central story, claiming he had mistaken Ms Steenkamp for an intruder that night, firing four bullets at his toilet door in a panic but adamant he had not intended to kill.
Mr Nel alleged numerous discrepancies in Mr Pistorius' version, highlighting that the Olympian was a stickler for detail on some matters and had a complete memory blank on others.
Mr Nel stepped Mr Pistorius through the minutiae of his version, citing numerous inconsistencies, contradictions and "tailoring" of evidence.
The athlete was also accused of using his tearful outbursts to avoid difficult questions and also changing his defence from one of self-defence, in which he genuinely believed he was acting lawfully, to another available – that of an "involuntary action".
By the time Mr Pistorius left the witness box, Mr Nel had presented a compelling picture of what the state alleges took place on the night Ms Steenkamp died based on the versions of "ear witnesses", ballistics and forensic experts and evidence found at the crime scene.
The state alleges that the couple had an argument, and the young model and law graduate packed her bag to leave. There was perhaps a tussle over her jeans, the only item not neatly folded in her gym bag, after which Ms Steenkamp fled to the bathroom in fear of her life as Mr Pistorius grabbed his weapon, likely following her screaming: "Get the f— out of my house.''
Ms Steenkamp locked herself in the toilet with her mobile phone, and the couple may have continued to argue for some time.
Eventually, Mr Pistorius opened fire, the first bullet striking Ms Steenkamp in the hip as she stood close to the closed bathroom door.
She screamed and fell backwards onto a wooden magazine rack. The second bullet missed, but ricocheted off a wall and hit her on her back. A third struck her right upper arm, the fourth entering her skull through her left hand, raised to her head in a protective position.
"You knew Reeva was behind the door and you shot at her. That's the only thing that makes sense," Mr Nel said in summary.
"She was talking to you – she was standing right in front of the bathroom door talking to you when you shot her [wasn't she?]"
"No she wasn't."
Mr Nel: "She wasn't scared of any intruder, she was scared of you."
"It's not true, my lady," Mr Pistorius replied in a small voice.
The court has now adjourned until May 5, when the defence will resume, calling about 10 more witnesses.
After that, Judge Thokozile Masipa is likely to adjourn for a further month to enable both sides to make their closing submissions in writing before final argument takes place.


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It's not the name on the bottle that matters with health and wellness buys, it's the stuff in the bottle. Stop paying more for brand names!
There's evidence that multivitamins aren't as useful as once thought. Instead, get your vitamins and minerals the old fashioned way — from these healthy, tasty foods.
Stop wasting your money on multivitamins that don't work, but do start following these other healthy habits that multivitamin buyers follow.
Herbal supplements can be great health boosters, but figuring out which ones are best can be difficult. Here are some tips on how to find out.
Everyone knows that milk is an excellent source of calcium, but not everyone drinks it. Here are other great ways to get calcium without the milk mustache.
The latest Dietary Guidelines for Americans identified a handful of vitamins and minerals Americans weren't getting enough of. What are these seven nutrients, exactly?
Not everything we eat gets absorbed and used by our body. Our cooking and digestive process destroys and degrades nutrients before our body can use it.
The health benefits of pomegranates are something like folk wisdom outside of the Western world, and like green tea, are only starting to seep into the American consumer's consciousness. [more]
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Product Information
Ariston 990363 Gasket - Return Group Aco 32 Mffi Lpg, Great deals on Ariston spares from Boiler Parts.
The Ariston Aco 32 Mffi Lpg Gaskets, O'rings, Seals and Washers are used to provide a seal to avoid a loss of air, gas and water in most appliances. Most of the O'rings, Gaskets, seals and Washers are specific to a certain placement within the boiler and are extremely seldom interchangeable owning to the size and thickness. They allow the system to be sealed to lessen the probability of leaking., Ariston Gaskets and Washers Can Be Delivered Next Day
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When replacing any spare part on appliances, use only genuine boiler spare parts that you can be assured conform to the safety and performance specification that the boiler manufacturer recommends. For your safety & your families, please do not use reconditioned or copy parts that have not been clearly authorised by the boiler manufacturer.


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