B lymphocyte differentiation in the rat: production and characterization of monoclonal antibodies to B lineage-associated antigens

Three mouse monoclonal antibodies (mAb) directed against rat B lineage antigens were produced. The mAb, designated HIS14 (IgG1), HIS22 (IgM) and HIS24 (IgG2b), were characterized for binding to lymphoid and nonlymphoid tissues by immunoperoxidase staining of frozen sections and by (double-) immunofluorescence staining of single cell suspensions from lymphoid organs. HIS14 recognized a pan B cell determinant: it reacted with virtually all cells of each anatomic B cell compartment and with about 95% of surface (s)Ig+ cells in thoracic duct lymph and in suspensions of spleen and lymph nodes. HIS22 and HIS24 detected B lineage-associated antigens expressed by major subpopulations of B cells. HIS22 predominantly stained the lymphocyte corona, but not (or weakly) the germinal centers and splenic marginal zones, whereas HIS24 reacted with both corona and germinal center and not (or weakly) with marginal zone. In accordance with this, substantial proportions of sIg+ cells in spleen cell suspensions did not express HIS22 or HIS24 determinants (20% and 27%, respectively). In bone marrow the vast majority of cytomplasmic mu+ pre-B cells were HIS14+ and HIS24+, and up to one third also HIS22+, indicating an appearance of the determinants early in B lymphocytopoiesis. The antigens recognized by HIS14, HIS22 and HIS24 are lost during the final stage of B cell differentiation: none of the mAb bound to plasma cells. As far as detectable, neither cells of myeloid and erythroid lineages in bone marrow nor thymocytes were stained by HIS14, HIS22, or HIS24. In suspensions of peripheral lymphoid organs (spleen and lymph nodes) but not in thoracic duct lymph, HIS14 and HIS24 labeled a small proportion (12% and 14%, respectively) of Ig- cells. HIS22 did not bind to Ig- peripheral lymphocytes. Reactivity of HIS14, HIS22 and HIS24 with nonlymphoid tissues was virtually absent; HIS22 stained the high endothelial venules in lymph nodes and Peyer's patches. As determined by immunoblotting, the antigenic determinants on lymph node cells recognized by HIS14, HIS22 and HIS24 were present on molecules with an apparent molecular mass of 205 kDa, 210 (and 175) kDa and 205 kDa, respectively, which is similar to the molecular mass of the B cell form of the rat leukocyte common antigen. In addition, the antigens recognized by HIS14, HIS22 and HIS24 co-capped with the leukocyte common antigen. This suggests that each of the three mAb recognize determinants present on the B cell form of the leukocyte common antigen.     

镁对在体兔缺血—再灌心脏的电生理效应

本实验通过结扎兔冠状动脉左室支复制动脉缺血－再灌注模型，应用心外膜接触电极记录单相动作电位，观察后除极电位在再灌注性心律失常中及镁离子的拮抗作用。结果表明，再灌性心律失常的５２．６％与早期后去极化有关。硫酸镁可终止及预防ＲＡ，对再灌中出现触发活动有抑制作用。     

Amino acid release from the cerebral cortex of the rabbit, its relationship with the electrocorticogram

The rate of release of five amino acids (alanine, glutamine, glutamic acid, glycine, GABA) from the surface of the sensorimotor cortex has been determined in rabbits under local anesthesia immobilized with gallamine. Concomitant recording of the electroencephalogram and of blood pressure has been carried out. The scope of the research was to determine if a difference in the release of these aminoacids could be correlated with variations in cerebral electrical activity. Two open-ended silver cylinders were adapted to the cortex and filled with saline. Every 10 min, the liquid was removed and assayed using a dansylating procedure followed by thin layer chromatographic separation and fluorimetric determination of the amino acids. The EEG of non treated animals, followed for a period of 100 min, remained synchronized for the majority of the recording; the biochemical determinations indicated a slight but constant fall in the rate of release of all the amino acids. A significant increase in the release accompanied the EEG desynchronisation induced either with eserine (0.2 mg/kg), amphetamine (2 mg/kg), or with electrical stimulation of the reticular formation.     

Anti-platelet autoantibodies from ITP patients recognize an epitope in GPIIb/IIIa deduced by complementary hydropathy.

Idiopathic thrombocytopenic purpura (ITP) is a frequent platelet disorder due to the presence of anti-platelet autoantibodies. Recently a fibronectin/fibrinogen receptor in platelets, integrin GPIIb/IIIa, has been implicated as the antigen in chronic ITP. To examine the epitopes involved in the autoimmune response against GPIIb/IIIa we have used concepts from the complementary hydropathy principle. We used the peptide Trp-Thr-Val-Pro-Thr-Ala, WTVPTA (deduced from the complementary nucleotide sequence to that which codes for the Arg-Gly-Asp, RGD, domain in fibronectin), to test the immunologic activity of ITP sera. Sera from 31 patients with clinically defined ITP were tested in ELISA for reactivity towards WTVPTA and affinity purified GPIIb/IIIa. Seventeen sera (57%) reacted strongly with the glycoprotein complex, five of which reacted with the peptide. By affinity chromatography of one of these sera, we were able to show that antibodies that bind to the peptide are within the population that binds to GPIIb/IIIa. Liquid phase competition experiments revealed that binding of ITP serum to WTVPTA was inhibited only by a hydropathically compatible peptide. Our data indicate that autoantibodies can bind to hydropathically generated antigenic determinants and thus, render these peptides clinically important as diagnostic tools.     

Theoretical investigation of estimation of steady and pulsatile blood flow and blood vessel cross section by CW NMR excitation

In this paper we show theoretically that when a magnetised blood bolus enters a CW NMR excitor coil of length Le at resonance and the signal from the T2-decaying, processing transverse magnetisation of the flowing blood spins is subsequently detected by a detector coil of length L separated from the excitor coil by a distance delta l, then by recording CW NMR signals at three positions such as delta l = 0, 0.5 and 1.0 cm one can eliminate the static tissue signal and measure non-invasively the steady component V0 as well as the total vessel cross section, beta accurately. The time dependent part of the CW NMR signal which depends on Vpulse(t), is also dependent on V0 non-linearly unless both L and Le are greater than 50 cm and delta l is zero. Finally, methods of obtaining true Vpulse(t) from the CW NMR signal after applying proper correction due to the steady flow are discussed.     

硫代乙酰胺所致急性肝衰时内毒素血症与肝内胆汁淤积关系的研究

本实验采用硫代乙酰胺（thioacetamide，TAA）灌胃复制大鼠急性肝功能衰竭的动物模型。检测血中内毒素、转氨酶、脂蛋白-x等含量以及胆汁流量。结果表明，大鼠发生了肝内胆汁淤积与内毒素血症（ETM），并且两者呈正相关（P＜0．05）。同时肝细胞膜上Na -K -ATP酶的活力明显降低，与ETM含量呈负相关（P＜0．05）。结果提示：ETM与肝内胆汁淤积的发生有密切关系．     

Levels of expression of CD19 and CD20 in chronic B cell leukaemias.

AIMS: To investigate whether the antigen levels of the B cell lineage markers CD19 and CD20 can distinguish between normal and neoplastic B cells or characterise distinct expression patterns among the chronic B cell leukaemias. METHODS: Peripheral blood cells from 70 patients with B cell disorders and 17 healthy donors were analysed by quantitative flow cytometry. Direct immunofluorescence staining was performed with phycoerythrin conjugated CD19 and CD20 monoclonal antibodies. Standard microbeads with different capacities to bind mouse immunoglobulins were used to convert the mean fluorescence intensity (MFI) values into number of antigen molecules/cell, expressed as antibody binding capacity (ABC). RESULTS: CD19 and CD20 ABC values in leukaemic B cells differed from those of normal blood B lymphocytes. The results identified distinct profiles of CD19 and CD20 expression in the various types of B cell leukaemias. In all leukaemias studied except hairy cell leukaemia (HCL), CD19 expression was significantly lower than the mean (SD) value in normal B cells (22 (7) x 10(3) molecules/cell), as follows: chronic lymphocytic leukaemia (CLL), 13 (7) x 10(3); B prolymphocytic leukaemia (B-PLL), 16 (9) x 10(3); splenic lymphoma with villous lymphocytes (SLVL), 15 (11) x 10(3); mantle cell lymphoma (MCL), 10 (7) x 10(3). In HCL there was strong CD19 expression (38 (16) x 10(3)). In contrast, the level of expression of membrane CD20 was higher than the mean (SD) value in normal B cells (94 (16) x 10(3) molecules/cell) in MCL (123 (51) x 10(3)); B-PLL (129 (47) x 10(3)); SLVL (167 (72) x 10(3)); and HCL (312 (110) x 10(3)); while it was significantly lower (65 (11) x 10(3)) in CLL compared with normal B cells and the other B cell leukaemias. CONCLUSIONS: Quantitative determination of CD19 and CD20 may provide useful diagnostic information for the study of B lymphoproliferative disorders.     

Osmoregulation of Hymenolepis microstoma

Summary The present report deals with a study of the osmoregulation of young Hymenolepis microstoma using the in vitro cultivation at different osmotic pressures: i.e. in eleven media with freezing point depressions ranging from 0.35 to 1.21° C. It has been shown that H. microstoma is able to develop optimally and equally well in media with freezing point depressions from 0.50 to 0.73° C. The worm seems to have a regulatory system operating successfully between these approximate limits. Media with lower or higher osmotic pressures do not cause death within the present experimental conditions but inhibit development gradually.It is concluded that, although cestodes are regarded as poikilosmotic, the present results with H. microstoma indicate that homoiosmotic properties might exist within a certain limited range of osmotic pressures.Supported by a N.A.T.O. grant.Encouragement by Prof. Dr. C. A. Hopkins is gratefully acknowledged.     

Individualised, micro CT-based finite element modelling as a tool for biomechanical analysis related to tissue engineering of bone

Load-bearing tissues, like bone, can be replaced by engineered tissues or tissue constructs. For the success of this treatment, a profound understanding is needed of the mechanical properties of both the native bone tissue and the construct. Also, the interaction between mechanical loading and bone regeneration and adaptation should be well understood. This paper demonstrates that microfocus computer tomography (microCT) based finite element modelling (FEM) can have an important contribution to the field of functional bone engineering as a biomechanical analysis tool to quantify the stress and strain state in native bone tissue and in tissue constructs. Its value is illustrated by two cases: (1) in vivo microCT-based FEM for the analysis of peri-implant bone adaptation and (2) design of biomechanically optimised bone scaffolds. The first case involves a combined animal experimental and numerical study, in which the peri-implant bone adaptive response is monitored by means of in vivo microCT scanning. In the second case microCT-based finite element models were created of native trabecular bone and bone scaffolds and a mechanical analysis of both structures was performed. Procedures to optimise the mechanical properties of bone scaffolds, in relation to those of native trabecular bone are discussed.     

Recurrent colonization of successively implanted tracheoesophageal vocal prostheses by a member of the Fusarium solani species complex

Tracheoesophageal vocal prostheses (TVP) in laryngectomized patients commonly deteriorate due to overgrowth by yeasts, particularly Candida species. We describe the first case of colonization of such devices by a member of the Fusarium solani species complex in a patient with a history of glottal carcinoma. Three isolates, from three prostheses, were found morphologically consistent with the traditional picture of F. solani. Ribosomal sequence analysis showed that the isolates belonged to a distinct, as yet apparently unnamed phylogenetic species within the F. solani species complex. This species, one of two distinct genetic types (genotype 2) traditionally considered part of the plant-pathogenic subtaxon Fusarium solani f. sp. radicicola, has not previously been identified as an agent of human or animal disease, although it is closely related to a known etiologic agent of mycetoma, an Acremonium-like species recently renamed Fusarium falciforme. Sequence and multisatellite M13 polymorphism analysis revealed no distinctions among the case isolates. Production of cyclosporine was detected for all three case isolates.