Amino acid sequence of a platelet-binding human anti-DNA monoclonal autoantibody

The complete amino acid sequences of the variable regions of the heavy and light chains of a human IgM monoclonal platelet-binding autoantibody have been determined. This antibody, HF2-1/17, produced by a human x human hybridoma prepared from lymphocytes of a patient with systemic lupus erythematosus and thrombocytopenia, is polyreactive with single-stranded DNA, synthetic polynucleotides, sulfated carbohydrates, and acidic glycolipids isolated from platelet membranes. The heavy chain is of the VHIII subgroup, and the light chain is of the VKI subgroup. The heavy chain is the expression product of the VH26 germline gene. The light chain bears significant homology to other immunoglobulins of known primary structure, including WEA, GAL, HAU, HK101, and DEE. These results suggest that HF2-1/17 may be an autoantibody derived with little or no modification from germline genes. A model of the antibody combining site suggests that arginine 24 and arginine 30 in the light chain (CDR1) interact with a surface defined by phosphate or sulfate groups of the antigen.     

Ontogeny of B-lymphocyte function. III. In vivo and in vitro studies on the ease of tolerance induction in B lymphocytes from fetal, neonatal, and adult mice

The ease of tolerance induction in B lymphocytes from fetal, neonatal, and adult mice was studied in vivo, in a cell transfer system, and in vitro. Three different tolerogens were used: ultracentrifuged BGG, DNP(6)-D-GL, and ultracentrifuged DNP(22)-BGG. Irradiated thymectomized mice were reconstituted with B cells from fetal or neonatal liver or adult spleen or bone marrow. The mice were injected with tolerogen 1 day later. They were given normal thymus cells and challenged with either BGG or DNP(44)-BGG between 4 and 14 days after tolerance induction. With BGG no difference in ease of B-cell tolerance induction was observed in mice reconstituted with B cells from 17-day fetal liver, neonatal liver, 8- day-old spleen, adult spleen, or adult bone marrow. B cells from 14-day fetal donors are relatively resistant to tolerance induction. In contrast, with DNP(6)-D-GL and DNP(22)-BGG B cells from neonatal donors were clearly more susceptible to tolerance induction than were B cells from adult donors. Comparable results were obtained in studies on tolerance induction in vitro. Neonatal B cells were more susceptible than adult B cells to tolerance induction upon culture with DNP(6)-D-GL or DNP(22)-BGG. However, neonatal and adult B cells were identical with respect to ease of tolerance induction in vitro with deaggregated BGG. The results suggest that there are multiple mechanisms for B-cell tolerance induction. Immature B cells appear to be more susceptible to tolerance induction by some mechanisms but not by others. It is suggested that immature B cells are more susceptible to tolerance induction with moderately polyvalent antigens such as hapten-carrier conjugates. With antigens like BGG which do not haverepeated epitopes no difference between mature and fetal B cells in regard to ease of tolerance induction is observed. These observations raise questions about the importance of relative ease of tolerance induction in immature B cells as a mechanism controlling the normal induction of self tolerance.     

Methodology for development of the Allergic Rhinitis and its Impact on Asthma guideline 2008 update

Background:  We describe the methodology for the 2008 update of the Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines. The methodology differs from the 2001 edition in several respects. The most prominent change is the application of the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach to compiling evidence, assessing the quality of evidence and grading of recommendations.
Methods and results:  Representatives of the GRADE working group joined the ARIA guideline panel to achieve these tasks. While most recommendations result from existing systematic reviews, systematic reviews were not always available and the panel compiled the best available evidence in evidence profiles without conducting actual reviews. The panel conducted two meetings and used the GRADE criteria to assess the quality of evidence (four categories of high, moderate, low and very low) and the strength of recommendation (strong and weak) based on weighing up the desirable and undesirable effects of management strategies, considering values and preferences influencing recommendations, and resource implications. The guideline panel has chosen the words 'we recommend'– for strong recommendations and 'we suggest'– for weak recommendations. Both categories indicate the best course of action for a given patient population, but their implementation, requires different considerations as we describe subsequently in this article.
Conclusions:  The 2008 update of the ARIA guidelines has become more evidence-based. Future iterations of the guidelines will further be improved by following the described processes even closer, such as ensuring availability of updated high quality systematic reviews for each question.     

Anti-proliferative and anti-remodelling effect of beclomethasone dipropionate, formoterol and salbutamol alone or in combination in primary human bronchial fibroblasts

Background:  Bronchial asthma is characterized by lower airway inflammation and remodelling. Anti-inflammatory treatment with inhaled corticosteroids (ICS) provides the mainstay of asthma therapy together with bronchodilation induced by short- and long-acting inhaled β₂-agonists. Lower airway fibroblasts may play a critical role in airway inflammation and remodelling, suggesting that they might represent an important target for the major anti-asthmatic drugs. The aim of our study was to investigate the effects of beclomethasone dipropionate (BDP), salbutamol and formoterol either alone or in combination on in vitro cultures of human bronchial fibroblasts.
Methods:  Fibroblasts were cultured in the presence of pro-inflammatory and proliferative stimuli, BDP, salbutamol and formoterol. The effects of drugs on cell proliferation were ascertained by ³H-thymidine incorporation. CD90 and CD44 expression were detected by flow cytometry and fibronectin secretion using the enzyme-linked immunosorbent assay technique.
Results:  This study showed that BDP alone has significant anti-proliferative effects on lung fibroblasts treated with basic fibroblast growth factor and the combination of BDP with formoterol or salbutamol strengthen these effects. Short-acting β₂-agonist (SABA) or long-acting β₂-agonist (LABA) by themselves did not show any significant effect on the different cultures. CD44 and CD90 expression and fibronectin production were modulated by pro-inflammatory and proliferative stimuli; the addition of the drugs brought them back near to the basal level.
Conclusions:  From this in vitro study, we can conclude that BDP, when combined with salbutamol or formoterol, exhibits enhanced anti-remodelling activity in bronchial fibroblasts, providing new insights on the additive effects of ICS and SABAs and LABAs for asthma therapy.     

生物降解型左旋18－甲基炔诺酮微球在大鼠的药代动力学

用放射免疫法研究了左旋１８－甲基炔诺酮（ＬＮＧ）微球和ＬＮＧ微晶在大鼠的药代动力学。大鼠单次ｉｍＬＮＧ微晶３５ｍｇ·ｋｇ－１后４．８８ｈ．血药峰值达６７．６６ｎｍｏｌ·Ｌ－１，ＭＲＴ为１０．１６ｄ，Ｔ＜０．３２ｎｍｏｌ·Ｌ－１为４１．５０ｄ；而单次ｉｍＬＮＧ微球２０．４，４１．１和８３．３ｍｇ·ｋｇ－１后，血药分别于６，４．６７和４．３３ｈ达峰浓度１５．１９，３３．６１和３８．５５ｎｍｏｌ·Ｌ－１，ＭＲＴ分别为６９．２３，６５．１２和６３．２５ｄ，Ｔ＜０．３ｎｍｏｌ·Ｌ－１分别为１６７．８１，１６９．７３和１６７．２３ｄ。可见ＬＮＧ微球在大鼠的ＭＲＴ和Ｔ＜０．３２ｎｍｏｌ·Ｌ－１分别约为ＬＮＧ微晶的６．６和４．２倍，提示该微球具有明显的缓释长效作用，且初始血药峰值明显低于肌注ＬＮＧ微晶。     

盐酸维拉帕米渗透泵片溶出度与人体生物利用度研究

溶出度按Weibull's分布处理得T_d=5.76 h,T₅₀=3.9 h,零级溶出速度常数K_t=9.9450,平均体外溶解时间MDT=5.391 h。测定8名健康受试者,单剂量口服,得C_max=76.2±16.7 ng/ml,T_amx=8.0 h,t_1/2=9.75 h,MRT=19.41 h,MAT=5.34 h,与Knoll公司SR片相比,F_rel=101.71%;与市售普通片相比,F_rel=96.16%。多剂量口服,得C_max=121.47±34.5 ng/ml,T_max=7.14 h。按Loo-Riegelman方程处理表明体内外显著相关。理论值与实测值基本相符。