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Cord Blood Transplantation and Biology
- Bryan Walsh
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1 Cord Blood Transplantation and Biology Hal E. Broxmeyer Introduction Three tissue sources of hematopoietic stem and progenitor cells are currently being used for transplantation purposes.1 The source which has been used the longest and for which we have the most information is adult bone marrow. Peripheral blood has also been used, but for the number of stem and progenitor cells from blood to be in high enough concentration to be of clinical efficiency, they need to be mobilized into the blood from the marrow after recovery from chemotherapy and/or after administration of growth factors. The most recent source of transplantable cells, until 1988 considered mainly discardable material, are those from the umbilical cord and placenta at the birth of a child. (2) This last source of cells is the focus of this brief review. Clinical Transplantation and Cord Blood Banking Since our original suggestion that cord blood could serve as a source of transplantable stem and progenitor cells, (2) which led to the first transplant using HLAmatched cord blood from a sibling (3) that successfully corrected the hematological and life-threatening manifestations of a genetic disorder, there have been well over 230 cord blood transplants done worldwide for an assortment of malignant and non-malignant disorders. (3-17) A listing of most of the disorders treated by sibling and unrelated cord blood transplantation is given in Table 1. This listing is not meant to be all inclusive as the author may not be aware of all the transplants done to date, and the rapidity with which cord blood is being used for transplantation suggests that new disorders will be evaluated with this source of stem/progenitor cells in the near future. Table 1: Disorders for which cord blood transplantation has been utilized as a treatment modality* Acute Lymphoid Leukemia - Remission, Relapse Acute Myeloid Leukemia - Remission, Relapse Chronic Myeloid Leukemia - Accelerated/Blastic Juvenile Chronic Myeloid Leukemia Myelodysplastic Syndrome Neuroblastoma Acute Megakaryocytic Thrombocytopenia Severe Combined Immunodeficiency Common Variable Immunodeficiency Wiskott-Aldrich Syndrome x-linked Lymphoproliferative Disease Hurler Syndrome Hunter Syndrome
2 Fanconi Anemia Severe Aplastic Anemia β-thalassemia Sickle Cell Anemia Lesch-Nyhan Syndrome Pure Red Cell Aplasia Osteopetrosis Globoid Cell Leukodystrophy Adrenoleukodystrophy Günther s Disease - Porphyrin Accumulation LAD Syndrome - Adhesive membrane Glycoprotein Deficiency Kostmann s Syndrome Diamond Blackfan * This is not meant to be an inclusive list. There may be other disorders treated by cord blood transplantation that the author is not aware of. The first transplant took place in October 1988, (3) and the child is well more than seven years after the transplant. That first cord blood transplant, as well as the first five done and a total of seven of the first ten transplants done, utilized cord blood that had been sent from a distant obstetrical unit to the author s laboratory, where they were tested for viability, number and proliferative capacity of granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitors, frozen, and stored cryopreserved until they were hand-delivered to the center where the transplants were done. (3-6) These centers included Paris, France; Cincinnati, Ohio; Baltimore, Maryland; and Minneapolis, Minnesota. The first cord blood bank was established in the author s laboratory. It was the later establishment of unrelated cord blood banks, in particular the one at the New York Blood Center from which a very large proportion of the unrelated cord blood transplant donor cells have come, that confirmed for unrelated cells (16,17) the low graft-versus-host disease (GVHD) originally noted with sibling cells (3-15) and that led to the first cord blood transplants for adults. While there have been a good number of clinical papers written on the experience of using sibling cells in the context of children as recipients,3-15 information has only recently begun to accumulate that relates the experience of centers doing transplants with unrelated cells for adult recipients who are of higher body weight. (16,17) Recipients of up to 80 Kg body weight have thus far been engrafted with unrelated donor cells, and patients in their thirties and forties have served as recipients with engraftment noted (J. Kurtzberg & P. Rubinstein, personal communications). The engraftment rates for sibling and unrelated cord blood transplants have been in the greater than ninety percent range, and GVHD, even using unrelated cells, has been relatively mild compared to use of similarly mismatched unrelated bone marrow donor cells. The GVHD seen in these circumstances has been, so far, easy to manage. This lowered amount of GVHD has not been associated with a higher level of relapse thus far when cord blood cells have been used to treat patients with leukemia. The rate at which cord blood has been used for transplantation has accelerated greatly. In 1988 and 1989, a total of only two cord blood transplants had been done. With
3 the growth and utilization of samples from the New York Blood Center Unrelated Cord Blood Bank, and the establishment of a number of other unrelated cord blood banks in the United States, Europe, Australia, Thailand, Japan and elsewhere, the rate of cord blood transplants will increase, as will our knowledge of the broadness of applicability of these cells. In addition to the above banks, a number of commercial ventures into the storage of cord blood for autologous or family use have opened up. It is the author s opinion that the development of the field of cord blood transplantation will be enhanced by the simultaneous development of banks that store donated bloods to be used in an unrelated setting, as well as banks that store cord blood for directed donations for related or autologous use. Both are important types of banks necessary to anticipate the needs of the future. Unrelated banks may not have the money, resources or inclination to store directed donations for an affected sibling or relative, and not all recipients in need of a transplant may find the appropriate HLA type of cord blood from an unrelated bank. As far as banking of autologous cells is concerned, the most perfectly matched set of cells for oneself is one s own cells. There are certainly enough births in the world to satisfy collections of cord blood for both unrelated and related/autologous banks. Most of the cord blood transplants done have utilized cryopreserved samples. While clinical experience with cryopreserved cord blood has been limited to use of cells frozen from weeks to a few years, theoretically, once cells have been frozen, and assuming no problems with the freezing methodology and with liquid nitrogen storage tanks, cells should remain in cryopreserved form for at least the lifetime of an individual. Our recent studies have demonstrated very high efficiency recovery of viable immature and mature subsets of CFU-GM, BFU-E and CFU-GEMM with high proliferative capacity from cord blood stored frozen for up to ten years.18 This recovery was similar to that of these same cells assessed after only a few weeks of storage. (2,19) There may still be information to be learned regarding the biophysics and biology of cryopreservation and efforts in this area of research could be rewarding. Cord blood stored frozen in a relatively unseparated form can be thawed and retrieved in viable form and further purified, transduced by retroviral vectors with new genetic material, and expanded ex vivo for more mature stem cells and for immature and mature subsets of progenitor cells. (20) Additionally, purified cord blood stem and progenitor cells can be frozen and recovered in viable form.21 Autologous cord blood transplants have thus far been done only in the context of gene therapy using non-cryopreserved cells. (22,23) Three patients, diagnosed in utero with adenosine deaminase (ADA)-deficient severe combined immunodeficiency (SCID), had their cord blood collected at birth, separated for CD34+ cells by column maneuvers, preincubated with a combination of growth factors and subsequently with retroviral vectors containing a normal ADA-gene, and then reinfused into the patients after a number of days. While it is too early to determine if this procedure has resulted in therapeutic benefit, it is clear that a small but significant number of the infused cells in these patients carry ADA-transduced genes. (22) The patients have been on pegylated ADA since birth, and most recently, decreasing the infusion of pegylated ADA has been coincident with a increase in the percentage and numbers of cells that express the transduced ADA-gene. (23)
4 Biology of Cord Blood Stem and Progenitor Cells Cord blood is enriched for hematopoietic stem and progenitor cells with high levels of proliferative, replating and expansion capacity in vitro. (2,19,20,24) These cells can be phenotypically characterized by cell surface markers as being CD34+++, CD38- and thy1+. These cells are mainly in a slowly cycling or resting condition with a prolonged G1 phase of the cell cycle and are insensitive to kill by pulse-treatment of the cells with high specific activity tritiated thymidine. However, these immature cells respond rapidly to stimulation of their proliferation with growth factors such as GM-CSF, G-CSF, IL-3 and erythropoietin in combination with potent co-stimulating molecules such as steel factor or the Flt3/Flk-2-ligand. Cord blood plasma is a rich source of known, as well as uncharacterized, growth factors, (25) but how these interrelate to regulate the growth of immature cord blood cells is not known. Well over 50 cytokines are now known to regulate the proliferation of hematopoietic stem and progenitor cells, at least in vitro; (26) some have direct action, others act indirectly by inducing the release of cytokines from accessory cells. Many of these cytokines have pleiotropic and what appear to be redundant activities. (26) It will probably be a while before we fully understand this interacting network of cytokine-cell interactions and which of these cytokines may be of more relevance physiologically. However, it is already clear that combinations of cytokines can be used to good advantage to expand hematopoietic stem and progenitor cells from cord blood ex vivo. (19,24,25) Unfortunately, whether or not those stem cells can be maintained or expanded by these ex vivo maneuvers has been hindered for all tissue sources of these earliest cells because no assay yet exists that can definitively recognize these long-term marrow repopulating stem cells from humans and quantitate them. The earliest human cell that can be quantitated is the long-term culture initiating cell, but it has not been demonstrated that this defines the long-term marrow repopulating cell. For this reason, a number of groups have begun using SCID mice, (27) transgenic SCID mice that express human GM-CSF, IL-3 and steel factor, (28) or non-obese diabetic (NOD) SCID mice (28) as possible in vivo models for detection and quantitation of candidate longterm marrow repopulating human stem cells. Immunology and Cord Blood Cells In order to understand what might be responsible for the apparently low levels of GVHD noted after cord blood transplantation, a number of groups (including our own) have begun to evaluate the immunological reactivity of cord blood lymphocytes and natural killer cells. (29-31) Laboratory studies that might, at least in part, explain lowered GVHD reflect lowered levels of cytotoxic T-cell activity generated from cord blood T- cells in response to primary, secondary and tertiary allogeneic cell stimulation (29) and the induction of a state of unresponsiveness of proliferation of cord blood T-cells after priming with alloantigen. (30) Of interest is that natural killer cell activity in cord blood is low, due to functional and phenotypic immaturity of these cells, but these cells do respond well to stimulation of killing activity by IL-2 and IL-12. (31) In-depth analysis of the mechanisms responsible for differences between adult and cord blood T-lymphocytes
5 and natural killer cells remains to be evaluated and could prove useful to our understanding of immune activity. Gene Transfer into Cord Blood Stem and Progenitor Cells Cord blood stem and progenitor cells are efficiently transduced with new genetic material by using either retroviral (32,33) or recombinant adeno-associated viral (AAV) vectors. (34) Much more information is known about the use of retroviral, compared with AAV, vectors, and it is still to be decided whether AAV vectors stably integrate into the genome, or if in fact this is mainly episomal and transient. In addition to adding genes to correct genetic disorders, a potential future use might be to add genes that code for cytokine or cytokine receptors to stem and progenitor cells. Genes for the erythropoietin- 35 and IL-9-36 receptors have been transduced into cord blood progenitors at the level of a single isolated CD34+++ cell in the absence and presence of combinations of growth factors including erythropoietin and IL-9. At this single cell level, enhanced detection of erythroid and multipotential colonies with an erythroid component was seen in cells transduced with the erythropoietin35- or IL-936-receptor genes when erythropoietin or IL-9 was added. This erythroid differentiation was even more apparent when cells were transduced with both receptor genes36 and low levels of erythropoietin and IL-9 were added to the wells. A future use for such transduction may be in the enhanced differentiation or expansion of certain more lineage-restricted cells in vivo or in vitro. However, whether this would be at the expense of self-renewal would have to be determined. Conclusion The field of cord blood transplantation is expanding rapidly. Cord blood banking is beginning to provide an addition or alternative to bone marrow registries. Currently, cord blood banks are extremely valuable when the appropriate HLA-types cannot be found in bone marrow registries or when a transplantation is needed immediately. Bone marrow registries can take months to identify a donor while banked cord blood is immediately available for use. While we have much to learn regarding the full range of the therapeutic benefits inherent in cord blood, it is anticipated that at some time in the not too distant future most individuals needing a transplant as treatment for a cancer or genetic disorder will be able to have one through either unrelated, related or autologous cord blood banks. Manipulation of cord blood cells for expansion, so that one sample will always be enough for one and perhaps numerous recipients seems possible; this possibly could be greatly accelerated if we had at hand a quantitative assay for the human longterm marrow repopulating stem cell. The correction of genetic defects by transfer of a new or normal gene is only just beginning to be evaluated clinically, and it is anticipated that cord blood will be a vehicle of choice for such correction in the future. Additionally, a greater understanding of gene transduction into early quiescent stem cells in cord blood may increase the possibilities for creative ideas for manipulation and modulation of the proliferative, self-renewal and differentiation of stem cells. The future for cord blood transplantation continues to look bright.
6 Acknowledgments: I thank Rebecca Miller for typing the manuscript. The author is supported by US Public Health Service Grants R01 HL56416, R01 HL54037, and a project in P01 HL53586 from the NHLBI of the NIH. Previous studies based on some of the work referred to herein were supported by grants R37 CA36464 and R01 HL46549 from the NCI and NIH. References 1. Lu L, Shen RN, Broxmeyer HE: Stem cells from bone marrow, umbilical cord blood and peripheral blood for clinical application: Current status and future applications. Crit Review Oncology/Hematol 22:61, Broxmeyer HE, Douglas GW, Hangoc G, Cooper S, Bard J, English D, Arny M, Thomas L, Boyse EA: Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells. Proc Natl Acad Sci USA 86:3828, Gluckman E, Broxmeyer HE, Auerbach AD, Friedman HS, Douglas GW, Devergie A, Esperou H, Thierry D, Socie G, Lehn P, Cooper S, English D, Kurtzberg J, Bard J, Boyse EA: Hematopoietic reconstitution in a patient with Fanconi s anemia by means of umbilical-cord blood from an HLA-identical sibling. New Engl J Med 321:1174, Wagner JE, Broxmeyer HE, Byrd RL, Zehnbauer B, Schmeckpeper B, Shah N, Griffin C, Emanuel PD, Zuckerman KS, Cooper S, Carow C, Bias W, Santos GW: Transplantation of umbilical cord blood after myeloablative therapy: analysis of engraftment. Blood 79:1874, Kohli-Kumar M, Shahidi NT, Broxmeyer HE, Masterson M, Delaat C, Sambrano J, Morris C, Auerbach AD, Harris RE: Haematopoietic stem/progenitor cell transplant in Fanconi anaemia using HLA-matched sibling umbilical cord blood cells. Brit J Haematol 85:419, Broxmeyer HE, Kurtzberg J, Gluckman E, Auerbach AD, Douglas G, Cooper S, Falkenburg JHF, Bard J, Boyse EA: Umbilical cord blood hematopoietic stem and repopulating cells in human clinical transplantation. Blood Cells 17:313, Bogdanic V, Nemet D, Kastelan A, Latin V, Petrovecki M, Brkljacic-Surlakovic L, Kerhin-Brkljacic V, Aurer I, Konja J, Mrsic M, Kalenic S, Labar B: Umbilical cord blood transplantation in a patient with Philadelphia chromosome-positive chronic myeloid leukemia. Transplantation 56:477, Vowels MR, Lam-Po-Tang R, Berdoukas V, Ford D, Thierry D, Purtilo D, Gluckman E: Brief Report: Correction of x-linked lymphoproliferative disease by transplantation of cord-blood stem cells. New Engl J Med 329:1623, Pahwa RN, Fleischer A, Than S, Good RA: Successful hematopoietic reconstitution with transplantation of erythrocyte-depleted allogeneic human umbilical cord blood cells in a child with leukemia. Proc Natl Acad Sci USA 91:4485, Kernan NA, Schroeder ML, Ciavarella D, Preti RA, Rubinstein P, O Reilly RJ: Umbilical cord blood infusion in a patient for correction of Wiskott Aldrich Syndrome. Blood Cells 20:245, 1994
7 11. Issaragrisil S, Visuthisakchai S, Suvatte V, Tanphaichitr VS, Chandanayingyong D, Schreiner T, Kanokpongsakdi S, Siritanaratkul N, Piankijagum A: Brief report: Transplantation of cord-blood stem cells into a patient with severe thalassemia. New Eng J Med 332:367, Kurtzberg J, Graham M, Casey J, Olsen J, Stevens C, Rubinstein P: The use of umbilical cord blood in mismatched related and unrelated hemopoietic stem cell transplantation. Blood Cells 20:275, Vilmer E, Sterkers G, Rahimy C, Denamur E, Elion J, Broyart A, Lescoeur B, Tiercy JM, Gerota J, Blot P: HLA mismatched cord blood transplantation in a patient with advanced leukemia. Transplantation 53:1128, Brichard B, Vermylen C, Ninane J, Cornu G: Persistence of fetal hemoglobin production of successful transplantation of cord blood stem cells in a patient with sickle cell anemia. J Ped 128:241, Wagner JE, Kernan NA, Steinbuch M, Broxmeyer HE, Gluckman E: Allogeneic sibling umbilical cord blood transplantation in forty-four children with malignant and non-malignant disease. Lancet 346:214, Kurtzberg J, Laughlin M, Graham ML, Smith C, Olson JF, Casey JR, Halperin ED, Ciocci G, Carrier C, Stevens CE, Rubinstein P: Single center transplantation of HLAmatched and partially mismatched unrelated placental bloodñan alternative source of hemopoietic stem cells for bone marrow transplantation. N Engl J Med, in press, Wagner JE, Rosenthal J, Sweetman R, Shu XO, Davies SM, Ramsay NKC, McGlave PB, Sender L, Cairo MS. Successful transplantation of HLA-matched and HLAmismatched umbilical cord blood from unrelated donors: analysis of engraftment and acute graft-versus-host-disease. N Engl J Med, in press, Broxmeyer HE, Cooper S: High recovery of immature hematopoietic progenitor cells with extensive proliferative capacity from human cord blood cryopreserved for 10 years. Clin and Exp Immunol, in press, Broxmeyer HE, Hangoc G, Cooper S, Ribeiro RC, Graves V, Yoder M, Wagner J, Vadhan-Raj S, Rubinstein P, Broun ER: Growth characteristics and expansion of human umbilical cord blood and estimation of its potential for transplantation of adults. Proc Natl Acad Sci USA 89:4109, Lu L, Ge Y, Li ZH, Freie B, Clapp DW, Broxmeyer HE: CD34+++ stem/progenitor cells purified from cryopreserved normal cord blood can be transduced with high efficiency by a retroviral vector and expanded ex vivo with stable integration and expression of Fanconi anemia complementation C gene. Cell Transplantation 4:493, Lu L, Li ZH, Broxmeyer HE: Recovery and characterization of CD34+ cord blood cells after cryopreservation. In Vivo, in press, Kohn DB, Weinberg KI, Nolta JA, Heiss LN, Lenarsky C, Crooks GM, Hanley ME, Annett G, Brooks JS, El-Khoureiy A, Lawrence K, Wells S, Moen RC, Bastian J, Williams-Herman DE, Elder M, Wara D, Bowen T, Hershfield MS, Mullen CA, Blaese RM, Parkman R: Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiency. Nature Medicine 10:1, 1995
8 23. Kohn DB, Weinberg KI, Lenarsky C, Crooks GM, Heiss LN, Nolta JA, Smogorzewska EM, Bastian J, Wara D, Elder M, Bowen T, Hershfield M, Blaese RM, Parkman R: Selective accumulation of ADA gene-transduced T-lymphocytes upon Peg-ADA dosage reduction after gene therapy with transduced CD34+ umbilical cord blood cells. Blood 86 (Suppl 1):295a, 1995 (abstr) 24. Broxmeyer HE: Cord blood, an alternative source for stem and progenitor cell transplantation. Current opinion in Pediatrics. Hematology and Oncology Section. Philadelphia, Current Science, 7:47, Ruggieri L, Heimfeld S, Broxmeyer HE: Cytokine-dependent ex vivo expansion of early subsets of CD34+ cord blood myeloid progenitors is enhanced by cord blood plasma, but expansion of the more mature subsets of progenitors is favored. Blood Cells 20:436, Broxmeyer HE: Commentary: Is interleukin-17, an inducible cytokine that stimulates production of other cytokines, merely a redundant player in a sea of other biomolecules? J Exp Med, in press, Vormoor J, Lapidot T, Pflumio F, Risdon G, Patterson B, Broxmeyer HE, Dick J: Immature human cord blood progenitors engraft and proliferate to high levels in immune-deficient SCID mice. Blood 83:2489, Bock TA, Orlic D, Dunbar CE, Broxmeyer HE, Bodine DM: Improved engraftment of human hematopoietic cells in severe combined immunodeficient (SCID) mice carrying human cytokine transgenes. J Exp Med 182:2037, Risdon G, Gaddy J, Stehman FB, Broxmeyer HE: Proliferative and cytotoxic responses of human cord blood T-lymphocytes following allogeneic stimulation. Cell Immunol 154:14, Risdon G, Gaddy J, Horie M, Broxmeyer HE: Alloantigen priming induces a state of unresponsiveness in human cord blood T cells. Proc Natl Acad Sci USA 92:2413, Gaddy J, Risdon G, Broxmeyer HE: Cord blood natural killer cells are functionally and phenotypically immature but readily respond to IL-2 and IL-12. J Interferon and Cytokine Res 15:527, Moritz T, Keller DC, Williams DA: Human cord blood cells as targets for gene transfer: potential use in genetic therapies of severe combined immunodeficiency disease. J Exp Med 178:529, Lu L, Xiao M, Clapp DW, Li ZH, Broxmeyer HE: High efficiency retroviral mediated gene transduction into single isolated immature and replatable CD34+++ hematopoietic stem/progenitor cells from human umbilical cord blood. J Exp Med 178:2089, Zhou SZ, Cooper S, Kang LY, Ruggieri L, Heimfeld S, Srivastava A, Broxmeyer HE: Adeno-associated virus 2-mediated high efficiency gene transfer into immature and mature subsets of hematopoietic progenitor cells in human umbilical cord blood. J Exp Med 179:1867, Lu L, Ge Y, Li ZH, Keeble W, Kabat D, Bagby GC, Broxmeyer HE, Hoatlin ME: Influence of retroviral-mediated gene transduction of recombinant human erythropoietin receptor gene into single hematopoietic stem/progenitor cells from human cord blood on the growth of these cells. Blood 87:525, 1996
9 36. Lu L, Li ZH, Xiao M, Broxmeyer HE: Influence of retroviral-mediated gene transduction of both the recombinant human erythropoietin receptor and interleukin-9 receptor genes into single CD34+++CD33- or low cord blood cells on cytokine stimulated erythroid colony formation. Exp Hematol 24:347, 1996
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Patient education in adolescents with congenital heart disease – does a computer-based program developed for adults work?
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Anette Johansson (-); Susanne Sloberg (-); Ewa-Lena Bratt (Institutionen för vårdvetenskap och hälsa)
Publicerad i:
XXXXIV Nordic Paediatric Cardiology Meeting in Umeå. 17-19 september 2014,
Konferensbidrag, övrigt
Sammanfattning (abstract):
Abstract Today about 90% of children with congenital heart disease (CHD) survive to adulthood and many need life-long medical follow-up. Structured transition programmes have the potential to improve continuation of cardiac follow-up and aim to enhance adolescents' and their families' knowledge of the medical condition; to augment adherence to medical regimens; to improve overall quality of life (QoL) and psychosocial status. Parents are initially responsible for their child´s health but should gradually hand over responsibility to the youth. Knowledge of the disease is an important and crucial part in taking responsibility for their health. The aim of this study was to describe how adolescents with CHD experience a computer-based patient education developed for an adult population with CHD. Methods: Individual structured interviews of adolescents with CHD. The text transcribed verbatim was analysed with qualitative content analysis. Results: The text analysis resulted in an overall theme "Increased overall awareness and knowledge in need of some adjustments" with three subcategories: "The program was easy to use but requires some adjustment", "Gave a greater knowledge and understanding about heart failure, treatment and lifestyle advice" and "an increased sense of connectedness" Conclusion: The computer program needs to be age adapted but was considered by most of the interviewed young patients as a good method to learn more about ones own heart condition. Some of the modules need to be reformulated to be simpler and more suitable for young people, but most felt that the program was easy to use.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
Postens nummer:
Posten skapad:
2014-09-25 12:00
Posten ändrad:
2014-09-26 22:24


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s4e12-Day of the Colossus.mkv
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I think I will just buy a bunch of laying type chicks and raise them myself. can't trust the people who sell "POL" pullets .


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Motorized Bicycle Elite Member
Join Date: Feb 2013
Location: Goose Creek, SC
Posts: 632
Re: New (old) stuff
So the 48 was completely disassembled before I put it on the bike because I didn't want to risk what happened to my first HT. The cylinder walls got scored badly because of leftover china trash inside from manufacturing. While I had it apart, I noticed that the transfers didn't match at the base, so I smoothed it out a little and matched the base to the cylinder. Not much metal got removed, just enough to clean it out and smooth the transition.
I also developed a small head gasket leak, so I lapped the head a little, raising compression just a touch.
The last 2 things I did was I fabbed a ghetto poopoo pipe because I got tired of smelling like 2stroke all the time. I think the poopoo pipe did something for performance because after that it ran terrible and I had to open the jet up a little bit to get rid of the high rpm WOT lean surges.
So overall, I don't really consider that much modification. So far though, the top speed has been clocked at 43 MPH. And that's on a 44t rag joint sprocket. It's got plenty of pickup too. It gets my 190 lb butt (plus 40-ish lb bike) up go speed in no time. I will put it against almost any modified 66 any day.
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In sept my gyno did blood test to check my hormones which included a prego test since I was late. At the time my level was negative 1. My period came 9/11 to 9/14; 10/8 to 10/11. In November, I feeling very nauseous...I took a home test the day before my expected period which was neg. Being 5 days late, I took a blood test and my HCG level was at 2.40. Next day I started to spot bright pink and later that night what I assume to be my period start very red...for the next 5 day than spotting for 3 extra days. My periods are 4 days plus that initial brown spotting in the beginning. Please tell me I'm not crazy...even my boyfriend thought I was pregnant without even knowing that I was late or that I was questioning it.
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I will be in Vienna for 3 nights (June 19, 20, 21). I am interested in seeing the muscial HAIR when I am there, but can not find too much info. on it except it is playing at that time. Any suggestions for website? I am interested in other theatre also. Any? Any suggestions on opera for a non-opera person. I want to go, but have never been to one before - suggestions? I appreciate any help.
Recent ActivityView all Europe activity »
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Vienna - HAIR (show)
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A good place to start:
Once you reserve your hotel, you could email the concierge and ask him/her to reserve you tickets to "Hair."
For an opera schedule, Vienna State Opera is www.wiener-staatsoper.at and the Volksoper, the second major opera house, is www.volksoper.at.
Have fun!
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P.S. Linda, it looks like Die Meistersinger von Nürnberg is on at the Vienna Staatsoper (State Opera House) in June -- it's accessible and fun for an opera beginner.
If you can find one, a Mozart opera like Marriage of Figaro or The Magic Flute, would also be a good first opera experience.
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Thanks for the suggestions.
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The official site for the musical is www.musicalvienna.at/hair
If you can read German, you can book tickets online through that site. However, if you want to arrange tickets through an English site, go to:
which will give you the details on dates, times, how to book tickets., etc.
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I saw Hair in Vienna! I am at work and don't have the info with me, but I will look when I get home. Our concierge got the tickets for us earlier on the day of the performance. Very decent seats. We took a cab to the theater, and had to go to a nearby hotel to call a cab for the return trip, but it was not a problem. Dress is casual but neat. The show was great. Not Broadway, but perhaps off-Broadway calibur. All the dialogue is in German, but if you know the story you won't have any problem following along. The songs are sung in English. Go for it! We also saw the Magic Flute Marionnette show at Shonbrunn. Also a treat.
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Thanks - this has been very helpful. Any other suggestions for entertainment?
Sign in to comment.


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(From track report)
Eugene Ford's Your Out, second by a neck to multiple graded stakes winner Summer Colony in the Delaware Handicap (gr. III) on July 21, is being shipped from those Mid-Atlantic environs to contest the $150,000 Arlington Matron (gr. III) on Labor Day."We put her on a van here this morning," said Your Out's trainer, Graham Motion, speaking over the phone from Delaware on Wednesday morning. "The Delaware Handicap was definitely a good effort for her," Motion said. "I knew she loved the surface at Delaware, but she was the local horse going against proven graded stakes winners, so her effort was somewhat of a surprise, although not all together so considering the way she had been running."In her previous start on June 22 in Delaware's $100,000 Obeah, the Maryland homebred had won going away by four lengths at the 1 1/8-mile Matron distance. Before that, Your Out had won Delaware's $55,000 Summer King by 3-1/2 lengths May 21."I don't think the shipping to Chicago will bother her," said Motion, who will arrive at Arlington in time to saddle Your Out in Monday's Matron. "I talked to Mike Gorham, who shipped to Arlington from here earlier in the summer, so we just followed the same routine that he did."I'm looking forward to coming to Arlington," said Motion. "It will be my first trip there, although Dave Bailey (Arlington racing secretary) has been trying to get me to come for several years."
Your Out Shipping In for Arlington Matron
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SNMP I/O Devices Make Monitoring Environmental Conditions Easy. Austin Lin Product Manager Wayne Chen Technical Service Moxa Inc.
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1 SNMP I/O Devices Make Monitoring Environmental Conditions Easy Austin Lin Product Manager Wayne Chen Technical Service Moxa Inc.
2 Overview According to the US Census Bureau s 2012 Statistical Abstract there were more than TWO MILLION cases of burglary and vandalism at unmanned remote sites from 2009 to 2012, with damage averaging at about US$2100 per incident. Some methods currently used to monitor unmanned remote sites are not very practical, and could result in potential threats being ignored. As the equipment used in sites have evolved and become more complex, it is not enough to simply monitor the equipment; we should also pay attention to aspects of the surrounding environment such as temperature, humidity, airflow, power, and smoke. To enhance onsite security and monitor equipment status at remote infrastructures, some IT administrators have used an alternative way for environmental monitoring that incorporates network management software (NMS) with a proprietary SCADA system. However, due to lack of functionality and interoperability with other systems, these solutions were costly and generated complex management implementations. Using a multitude of management systems and network-related commands made managing and maintaining these systems extremely difficult. In addition, IT administrators also needed to take the time to learn a totally different protocol such as Modbus. Released on October 1, 2012, all rights reserved. Moxa manufactures one of the world s leading brands of device networking solutions. Products include industrial embedded computers, industrial Ethernet switches, serial device servers, multiport serial boards, embedded device servers, and remote I/O solutions. Our products are key components of many networking applications, including industrial automation, manufacturing, POS, and medical treatment facilities. How to contact Moxa Tel: Fax: Web: Copyright Moxa Inc. 1
3 What is SNMP and why use SNMP for environmental monitoring? This paper will first explain what SNMP is, why you should use SNMP for environmental monitoring, and then discuss how an SNMP environmental monitoring device will help ease the management and implementation process. Lastly, we will give a real life application and then address how Moxa s SNMP supported I/O devices will help you easily set up your environmental monitoring system. SNMP is a standard application protocol used for collecting information or configuring network devices such as routers, switches, servers, and environmental sensors (for temperature, humidity, etc.). Today, the third generation of SNMP is the standard for network management. An SNMP-enabled network consists of a Network Management Station (NMS), agent, and Management Information Base (MIB). For the NMS to communicate with its agents, each SNMP agent will include its own MIB file, which is a text file that includes a collection of all the managed objects. The MIB file defines a set of hierarchically organized characteristics associated with the managed objects, such as the object identifier (OID), access right, and data type of the objects. After the MIB compiler converts these text-based MIB modules into a format usable by the NMS, the NMS can start to read or write the managed device using different commands to obtain device-specific information. The NMS uses three basic delivery mechanisms to exchange information with an SNMP agent: First is the GET command, which allows the manager to request information for a specific list of objects. Second is the SET command, which allows the manager to request a change in the value of a specific variable. Lastly, the SNMP TRAP allows the agent to inform the manager of significant events. Most of the messages are issued only by the NMS. Only trap messages are initiated by the agent, and are typically used to notify the NMS of an alarm condition before the NMS queries that object. Since SNMP was first developed, three versions have been introduced by the Internet Engineering Task Force (IETF). SNMPv1 provides the bare minimum of network management functions and processes required to define network management information; however, it has security defects. SNMPv2 is compatible with SNMPv1, has extended features such as the ability to retrieve data in bulk rather than individually, allows the NMS to acknowledge trap message, supports other data types such as Counter 32, and provides various error codes, allowing it to distinguish errors in greater detail. SNMPv3 goes even further by encrypting the communication strings used to access agents and the SNMP data transmitted between devices. 2
4 Why use SNMP for environmental monitoring? Over the last decade, SNMP environmental monitoring has become critical for any company with a large network. As any IT company can tell you, it is hard for network administrators to remotely manage a large network that is not confined to a single area. Most of the time, these sites are isolated and not conveniently accessed or reached by foot, making it impractical to station a person onsite to monitor everything that happens. SNMP environmental monitoring employs the Master-Agent concept to transmit messages between the central alarm master and its agents at each network site. Furthermore, SNMP protocol is a familiar IT protocol for network management. By enabling SNMP for environmental monitoring, the IT administrator won t need to take the time to learn another protocol and can easily monitor not only the network but the environment of the system. What are the benefits of SNMP environmental monitoring devices? As the world continues to advance and business efficiency becomes more and more dependent on connected computer systems, ensuring the reliability, performance, and security of the monitoring devices has become absolutely necessary. What are the benefits of using SNMP environmental monitoring devices? Cost savings and easy integration with an existing NMS Normally, integrating a non-it system into an IT system will increase the overhead on processing and network elements, but by using an SNMP-supported environmental monitoring device, IT administrators can quickly and painlessly integrate these SNMPsupported devices into their existing NMS. And because it uses the same protocol, IT administrators would not need to spend much time to familiarize themselves with network monitoring extensions after integration, and the process will not increase the amount of overhead. Integrating the environmental monitoring system under a single SNMP platform will not only enable growth and platform independence, but will also allow the entire system to be controlled by a single, centralized NMS. While larger IP networks may require SNMP proxies to minimize WAN-based SNMP traffic, the reduction in the number of NMSs compared to other alternatives will enable IT to focus on other issues and to simplify the management network. Fast response with event-driven monitoring system for your assets Implementing an effective environmental monitoring system to protect these remote sites will drastically reduce the workload of the administrator to manually monitor the condition at individual remote site. SNMP trap is event driven, allowing the administrator to custom define alarm traps, which the administrators can respond to immediately in response to abnormal or critical events. The ability of the system to inform an NMS, and ultimately the administrator, of the safety conditions, will result in an increase in remote site safety and reduction in system failure. 3
5 Privacy/security issues resolved with SNMPv3 encryption By and large, administrators need a functioning network management protocol even when major portions of the network and its services are impaired. With SNMPv3 s incorporated security and administration mechanisms, SNMPv3 is able to work independently of any other network services for the secure transmission of SNMP messages. With SNMPv3, administrators are protected from the following threats: Masquerade (data origin authentication): interloper assumes the identity of a sender to gain its privileges. Modification of information (data integrity): alteration of in-transit messages. Modification of message stream: messages are re-ordered, delayed, or replayed Disclosure (data confidentiality): privileged information is obtained via eavesdropping on messages. Data from SNMP devices can be collected securely without fear of tampering or corruption. Confidential information such as SNMP Set command packets that change a router s configuration can be encrypted to prevent the information from being exposed on the network. 4
6 Moxa s Click&Go offers value-added user-control capability In addition to these benefits, Moxa also offers its patented programming tool Click&Go. Moxa s Click&Go control logic grants administrators unparalleled control over the behavior of their I/O devices, without involving third-party development tools or complex code. With Moxa s Click&Go, various user-defined events can be configured through the intuitive IF-THEN-ELSE logic for local control to send various types of alarms, including TCP/UDP messages, s, SMS messages, and SNMP traps. For example, to configure a fire alarm, just select IF DI fire alarm is ON THEN DO buzzer ON ELSE DO buzzer OFF. And then click, click, click, you re done. Programming-free Click&Go I/O Control Logic The traditional disadvantage of using SNMP traps is the limitation of the information conveyed. With only seven standard trap numbers available in SNMP, it can be difficult to tell what is happening just by looking at the traps. Moxa s Click&Go gives the users the flexibility of defining SNMP trap formats, which allows variable binding, and adds more information to each SNMP trap. For example, an SNMP trap that is triggered when a door sensor is tripped can include the message Intrusion Alert, plus date, time, and server name. User Definable SNMP Trap Content 5
7 Real life application: ETC system on a highway Both for environmental reasons and to control traffic jams, countries all over the world are implementing unmanned ETC systems throughout the major highways in their cities. These tollgates are equipped with laser detectors to monitor vehicles by identifying the size of the vehicle, and generating a pulse output to trigger different cameras to take snapshots. The ETC tollgates help regulate highway traffic by smoothing out the traffic flow and reducing gas emissions. ETC Tollgate System Thousands of ETC tollgates are being installed on highways around the globe, and to help with monitoring and maintaining the tollgates, some system integrators are choosing Moxa s products. The iologik E2242, for example, is an environmental monitoring device that can monitor ETC tollgates via the Ethernet. In addition, the iologik E2242 provides local control capability, an AI channel to detect water leaks, electrical conditions, and monitor the status of air conditioners, and uses an RS-485 interface for temperature/sensor monitoring. Furthermore, Moxa s Click&Go logic offers a straightforward if-then-else local control capability, giving users the flexibility to define different traps to customize their front-end control capability. 6
8 Moxa s Intelligent SNMP Solutions Moxa s iologik E1200 (non-programmable), E2200, E4200, and W5300 series of intelligent SNMP environmental monitoring devices allow you to easily monitor your telecommunication infrastructure environment and remote sites. These devices are specifically designed for information field applications that use the SNMP protocol for monitoring environmental conditions (temperature, humidity, airflow, door, and intrusion) with programming capability and multiple alarm notifications that include , SMS text messaging, and SNMP traps. Disclaimer This document is provided for information purposes only, and the contents hereof are subject to change without notice. This document is not warranted to be error-free, nor subject to any other warranties or conditions, whether expressed orally or implied by law, including implied warranties and conditions of merchantability, or fitness for a particular purpose. We specifically disclaim any liability with respect to this document and no contractual obligations are formed either directly or indirectly by this document. 7
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MOSCOW, January 19. /TASS/. The situation in Syria, in Ukraine, the North Korean nuclear problem, as well as a number of bilateral issues will be in focus of a meeting between Russian Foreign Minister Sergey Lavrov and US Secretary of State John Kerry in Switzerland's Zurich on January 20, Russian Deputy Foreign Minister Mikhail Bogdanov told TASS on Tuesday.
"Lavrov and Kerry will discuss bilateral relations between Russian and the United States, Syria, Ukraine, North Korea and other subjects," he said. "There are many issues of mutual interest for Washington and Moscow. I' like to remind that the ministers are meeting as was agreed by the two heads of state."
"As for Syria, the ministers will discuss all issues of the Syrian settlement in line with [United Nations Security Council] resolution 2254," Bogdanov said. "They include the launch of intra-Syrian talks, anti-terror efforts, the political process, the humanitarian situation. They will exchange opinions on lists of terrorists."
"And the most importan thing now: if we are committed to begin intra-Syrian talks on January 25, we must seek to settle the outstanding problems," he said. "We hope we will be able to meet the deadline since we must fulfil the resolution [United Nations Security Council resolution 2254]. It says that the talks are to begin in January."


text: Entwined Dragon Claws 77,227pages on this wiki Add New Page Edit VisualEditor History Add New Page Talk0 In-Game Examine Window Entwined Dragon Claws other resources EQ2U · LootDB · EQ2LL · ZAM · Census xml Entwined Dragon Claws FABLED LORE NO-TRADE EVIL ONLY +18 str +23 agi +10 sta +130 health +7 Crushing Mark of the Scorn FistsOne-Handed Crushing Damage 24 - 72 Slashing (56.8 Rating) Delay 1.7 seconds Level 70 (Tier 8) Bruiser, Monk Effects: When Equipped: On a successful attack this spell has a chance to cast Mark of the Scorn on target of attack. This effect will trigger an average of 1.8 times per minute. Inflicts 369 - 616 magic damage on target Obtain: [How can I get one?] try LootDB or the linkback What does this information mean? Retrieved from "http://eq2.wikia.com/wiki/Entwined_Dragon_Claws?oldid=448599" Categories: Equipment Fists (Weapon Type) One-Handed Crushing (Equipment Type) Primary (Inventory Slot) Equipment pages with existing default images Fabled Equipment Crushing (Equipment) Tier 8 Equipment Brawler Equipment Bruiser Equipment Monk Equipment Articles needing obtain information Articles missing itemlink Add category Cancel Save


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Two Opening Repertoires for White: Vol 2: Aggressive Plans for White - Hardinge Simpole chess classics (Paperback)Raymond Keene, Byron Jacobs
Paperback Published: 21/11/2003
- Can be ordered from our supplier
A companion volume to Two Opening Repertoires for White: Volume 1, this book details some more aggressive strategies in the opening for White again based on the key move 1d4! Together the two volumes give ambitious White players a complete arsenal of choice against all Black defences. The companion volume is Two Opening Repertoires for White: Volume 1: Keeping control with White
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