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991.
992.
Lymphocyte predominance Hodgkin's disease: lineage and clonality determination using a single-cell assay 总被引:4,自引:1,他引:4
Lymphocyte predominance Hodgkin's disease (LPHD) is a clinically indolent condition. Although there is evidence that the putative neoplastic cell in this disease, the "L&H" cell, is of B-cell lineage, there is conflicting data concerning the clonality of these cells. Our study was aimed at clarifying the issue of lineage and clonality of the L&H cells of LPHD using a single-cell assay. Four cases of LPHD were studied. To circumvent the difficulties of obtaining fresh tissue and to be able to study representative cases, a new method was developed to obtain single-cell suspensions of L&H cells from archival formalin- fixed paraffin-embedded tissue. Single L&H cells were identified by morphology and immunostaining for epithelial membrane antigen, isolated using a micropipette, and subjected to polymerase chain reaction (PCR) amplification of the complematarity determining region 3 (CDR3) of the Ig heavy chain (IgH) gene, which is B-cell clone-specific. The PCR products were size-fractionated by polyacrylamide gel electrophoresis and representative products were directly sequenced. Single T cells and small B cells were also isolated from the tissues and used as negative and positive controls, respectively. In all four cases of LPHD, the IgH CDR3 of single L&H cells could be amplified. Within each case, the IgH CDR3 of single L&H cells was found to be of different length or of different sequence. Therefore, our results provide strong evidence for the B-cell origin of the L&H cells and the polyclonal nature of LPHD. 相似文献
993.
The present study was undertaken to characterize the species specificity and diversity of lactogenic and somatogenic binding to serum among mammals and to classify GH-binding protein (GH-BP) in this group of species. Animal sera were characterized on the basis of human (h) GH and bovine (b) PRL binding levels, binding specificity toward GHs and PRLs, binding affinity constant (Ka), and the ability of a monoclonal antirabbit GH receptor antibody (MAb-7) to inhibit the binding to serum. Analyses of the results yielded a classification of the mammalian sera into five types of GH binding, which we elected to call GH-BP. The guinea pig had undetectable levels of GH-BP and was labeled type 0. Type I GH-BP was found in the mouse and the rat. hGH binding to GH-BP of type I serum was of low affinity (1.2-3.9 x 10(8) M-1) with high IC50 (approximately 100 ng/tube) and was purely somatogenic in nature. Type II GH-BP was found in the goat, sheep, and cow. Their IC50 of hGH binding was approximately 4-fold lower than that of type I GH-BP. Their binding of hGH was relatively specific and only marginally displaced by the PRLs. Type III GH-BP was found in the rabbit, horse, cat, dog, and pig. These animals' sera had high affinity binding toward hGH (4.7-9.2 x 10(9) M-1), with low IC50 (approximately 2 ng/tube) and dominant lactogenic binding. The great similarity of type III GH-BPs was further stressed by the ability of MAb-7 to inhibit [125I]hGH binding to all type III sera, but not to the other mammalian sera tested. Type IV GH-BP was found in the human and rhesus monkey sera. These were characterized by binding affinities that were intermediate between those for types II and III GH-BP and by the inability of nonprimate GHs to compete with hGH binding. To directly confirm the potent effect of the lactogenic hormones in type III GH-BP, the specific binding of bPRL to sera of type III animals was studied. [125I]bPRL-specific binding was determined under optimized assay conditions and was the highest in rabbit serum, with the following rank order being observed: rabbit > horse = dog = pig > cat. Scatchard analysis of [125I]bPRL binding revealed linear plots with similar affinity constants (Ka) of 1.7-3.3 x 10(9) M-1.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
994.
995.
Biochemical, immunological, and in vivo functional characterization of B-domain-deleted factor VIII 总被引:12,自引:4,他引:12
Coagulation factor VIII (FVIII) is a cofactor in the intrinsic pathway of blood coagulation for which deficiency results in the bleeding disorder hemophilia A. FVIII contains a domain structure of A1-A2-B-A3- C1-C2 of which the B domain is dispensable for procoagulant activity in vitro. In this report, we compare the properties of B-domain-deleted FVIII (residues 760 through 1639, designated LA-VIII) to wildtype recombinant FVIII. In transfected Chinese hamster ovary (CHO) cells, LA- VIII was expressed at a 10- to 20-fold greater level compared with wildtype FVIII. The specific activity of purified LA-VIII was indistinguishable from wild-type recombinant FVIII and both exhibited similar thrombin activation coefficients. Wildtype recombinant-derived FVIII and LA-VIII also displayed similar timecourses of thrombin activation and heavy chain cleavage. However, compared with wildtype recombinant-derived FVIII, the light chain of LA-VIII was cleaved fivefold more rapidly by thrombin. Addition of purified von Willebrand factor (vWF) did not alter the kinetics of thrombin cleavage or activation of either wildtype recombinant-derived FVIII or LA-VIII. The immunogenicity of LA-VIII was compared with wildtype FVIII in a novel model of neonatal tolerance induction in mice. The results did not detect any immunologic differences between wildtype FVIII and LA-VIII, suggesting that LA-VIII does not contain significant new epitopes that are absent in wildtype FVIII. LA-VIII was tolerated well on infusion into FVIII-deficient dogs and was able to correct the cuticle bleeding time similar to wildtype recombinant factor VIII. In vivo, LA-VIII was bound to canine vWF and exhibited a half-life similar to wildtype recombinant FVIII. These studies support that B-domain-deleted FVIII may be efficacious in treatment of hemophilia A in humans. 相似文献
996.
Anti-My-28, an antigranulocyte mouse monoclonal antibody, binds to a sugar sequence in lacto-N-neotetraose 总被引:5,自引:0,他引:5
Spitalnik SL; Schwartz JF; Magnani JL; Roberts DD; Spitalnik PF; Civin CI; Ginsburg V 《Blood》1985,66(2):319-326
Anti-My-28 is an IgM kappa monoclonal antibody produced by a hybridoma prepared from spleen cells of a mouse immunized with normal human granulocytes. By immunofluorescence it binds to human granulocytes but not to monocytes and lymphocytes. However, after treating cells with neuraminidase, the antibody also binds to lymphocytes and monocytes and to many leukemic cell lines and patient leukemic blast cells. Anti-My- 28 binds to several neutral glycolipids and desialylated gangliosides of leukocytes and erythrocytes as detected by radioimmunoassay and immunostaining of thin-layer chromatograms. It recognizes a sugar sequence in lacto-N-neotetraose, Gal beta 1-4GlcNAc beta 1-3Gal beta 1- 4Glc. This tetrasaccharide occurs in the glycolipids paragloboside and sialosylparagloboside, and its distal trisaccharide sequence is found in higher glycolipids and in glycoproteins. 相似文献
997.
998.
Carlos Antonio Negrato Renan M Montenegro Jr Rosiane Mattar Lenita Zajdenverg Rossana PV Francisco Belmiro Gonçalves Pereira Mauro Sancovski Maria Regina Torloni Sergio A Dib Celeste E Viggiano Airton Golbert Elaine CD Moisés Maria Isabel Favaro Iracema MP Calderon Sonia Fusaro Valeria DD Piliakas José Petronio L Dias Marilia B Gomes Lois Jovanovic 《Diabetology & metabolic syndrome》2010,2(1):1-14
There is an urgent need to find consensus on screening, diagnosing and treating all degrees of DYSGLYCEMIA that may occur during pregnancies in Brazil, considering that many cases of DYSGLYCEMIA in pregnant women are currently not diagnosed, leading to maternal and fetal complications. For this reason the Brazilian Diabetes Society (SBD) and the Brazilian Federation of Gynecology and Obstetrics Societies (FEBRASGO), got together to introduce this proposal. We present here a joint consensus regarding the standardization of clinical management for pregnant women with any degree of Dysglycemia, on the basis of current information, to improve medical assistance and to avoid related complications of Dysglycemia in pregnancy to the mother and the fetus. This consensus aims to standardize the diagnosis among general practitioners, endocrinologists and obstetricians allowing the dissemination of information in basic health units, public and private services, that are responsible for screening, diagnosing and treating disglycemic pregnant patients. 相似文献
999.
Nassar AA Childs RD Boyle ME Jameson KA Fowke M Waters KR Hovan MJ Cook CB 《Journal of diabetes science and technology》2010,4(5):1156-1163
Background
This study aims to identify self-management strategies used by patients with diabetes in hot weather, examine knowledge of safe temperatures and exposure times, and evaluate comprehension of weather data.Methods
Patients attending an endocrinology clinic in southwestern United States were surveyed.Results
One hundred fifty-two surveys completed from November 30 to December 31, 2009 werMean patient age was 64 years; diabetes duration e analyzed 15 years; 51% were women 83% had type 2 diabetes, 58% were non Hispanic white, and 77% were on insulin injections. Mean hemoglobin A1c was 7.9%; 38% had hemoglobin A1c values of 8.0% or higher. Patients used an array of personal protective measures during hot weather, and 71% limited heat exposure to less than one hour. Respondents typically took steps to protect their diabetes medication and supplies, but 37% chose to leave medications and supplies at home. Although 73% of respondents had received information regarding the effect of heat on insulin, only 39% had received information about the effect of heat on oral medications, 41% on glucose meters, and 38% on glucose test strips. The temperature at which patients took protective measures varied. Only 55% knew the definition of the heat index.Conclusion
Most survey participants took measures to protect themselves from the heat, although gaps in knowledge were evident. Many patients had suboptimal glycemic control, placing them potentially at risk for dehydration during the hottest months. Increased public awareness of this topic is needed, and diabetes education programs should include information about the heat where appropriate. 相似文献1000.
Joshua A. Waters Ray Chihara Jose Moreno Bruce W. Robb Eric A. Wiebke Virgilio V. George 《JSLS, Journal of the Society of Laparoendoscopic Surgeons》2010,14(3):325-331