Pst I RFLP, revealed with DQ and DQ probes, was compared with Taq I RFLP using a panel of DR-homozygous cell lines and HLA-typed family members. Taq I patterns, characteristic for each DR-associated DQ and allelic forms, were recognized in the homozygous state and then proven to segregate in the heterozygous members of informative families. The presence of both specific and chains was found to be necessary to form the type of DQ molecule specifically recognized by two alloreactive T-cell clones. Particular and associations also seem to be responsible for some Dw splits of the DRw6-positive cells. Taq I RFLP analysis may be more complex than the Pst I analysis, but is certainly more informative and complete, considering the type of information we were seeking by performing these types of experiments.Abbreviations used in this paper BSA
bovine serum albumin
- GLO
glyoxalase
- kb
kilobase(s)
- LCL
lymphoblastoid cell line
- MHC
major histocompatibility complex
- PBL
peripheral blood lymphocyte
- PLT
primed lymphocyte test
- RFLP
restriction fragment length polymorphism
- SDS
sodium dodecyl sulfate
- SSC
standard sodium citrate
- SSCP
sodium, sodium citrate, sodium phosphate
- TBE
Tris-borate, boric acid, ethylenediaminetetraacetate (EDTA)
- TCGF
T-cell growth factor 相似文献
The interactions of alloreactive T lymphocytes with the vascular endothelium were studied in an in vitro model of lymphocyte adherence to cultured human arterial endothelial cell (HAEC) monolayers. Donor-primed lymphocytes (DPL) were shown to have significantly greater adherence to donor HAEC than were third-party primed lymphocytes. Limiting dilution analysis of adherent DPL showed an enrichment of donor-reactive lymphocytes compared with nonadherent DPL. This study examines the allospecific nature of this increased lymphocyte adherence. HAEC constitutively express class I HLA Ag and can be induced by IFN-gamma to express class II Ag. DPL adherence to class I+ HAEC was inhibited only in the presence of mAb directed against class I Ag. DPL adherence to class I+ and class II+ HAEC was inhibited in the presence of mAb directed against class I and class II Ag. Class I- and class II-specific adherence was also shown to involve CD8 and CD4 molecules, respectively, whereas lymphocyte function-associated Ag do not appear to play a major role in long term alloreactive lymphocyte adherence to HAEC. These findings suggest that alloreactive lymphocyte adherence to HAEC is mediated by two mechanisms. One is based on allorecognition, primarily of HLA Ag, and the other is related to presumably non-Ag-specific interactions between activated lymphocytes and the vascular endothelium. The studies presented provide evidence to suggest that HLA-specific lymphocyte adherence to endothelium may significantly contribute to the development of alloreactive lymphocyte infiltrates within the allograft. 相似文献
Pseudohypoparathyroidism type Ia (PHP-Ia) is a hereditary disease characterized by resistance to PTH and other hormones that act via cAMP. Patients have deficient activity of Gs, the subunit of the G protein, which couples hormone receptors to stimulation of adenylate cyclase. We describe two new mutations discovered in two sporadic patients with PHP-Ia. Using genomic DNA, we have amplified exons 2–13 of the Gs gene (GNAS1) by PCR, and sequenced the resulting products. Both patients had Albright's hereditary osteodystrophy, resistance to multiple hormones, and deficient Gs activity. In the first patient, a deletion of a C in exon 5 at codon 115 was found. In the second patient, an insertion of a C in exon 10 at codon 267 was detected. Both these heterozygous mutations cause frameshift, and predict decreased production of Gs. This report adds two new Gs mutations to the known ten mutations recently described. 相似文献
Novel backbone-to-side chain and backbone-to-backbone cyclic analogues of substance P (SP) were prepared by solid-phase synthesis and screened for biological activity. An analogue containing a thioether- lactam ring between positions 9 and 11 showed an EC50 value of 20nM toward the neurokinin 1 (NK-1) and was inactive toward the NK-2 and NK-3 receptors. On the other hand, in a multiple backbone cyclic peptide library of similar analogues, in which the sulphur was excluded from the ring, very low activity was detected. The activity was re-evaluated and was found to be even lower (EC50=0.11 mM ) than the previously published data. These results indicate that the thioether moiety has a crucial role in receptor activation. The results also show tolerance of the NK-1 receptor, but not NK-2 or NK-3, to cyclization of the C-terminal portion of the SP6–11 hexapeptide. 相似文献
In order to determine the suitability of thecyprinodont fish Aphanius dispar as a biologicalcontrol agent for mosquito larvae, the effects oftemperature, salinity, feeding rate and photoperiod onits reproductive condition were studied. The factorsexamined were the gonado-somatic index (GSI) and fourdistinct stages of cell maturation in the ovaries(oogonia, primary oocytes, secondary oocytes andmature ova). The fish were found to be tolerant to awide range of the examined factors. GSI's were notaffected by ambient temperatures over the range of18 °C to 37 °C. However, oocyte maturationstages increased as temperatures rose from 18 °Cto 27 °C, with mature ova first being found at thelatter temperature. No significant differences amongmaturation stages were found between27 °C–37 °C. At salinities ranging from of0 ppt to 56 ppt, no significant differences were found betweenmean GSI's, except for the two extremes of the testrange. The ovaries of fish kept in distilled watercontained primordial germ cells only. In feedingexperiments, significant differences were found onlybetween the ovaries (GSI's and oocyte maturationstages) of fish deprived of food and those fed at1% body weight per day. Regarding photoperiod,a decrease in day length from 14L:10D (the longest dayof the year) to 10L:14D (the shortest) caused adecrease in oocyte maturation stages. Fish at theintermediate level (12L:12D) still had ovariessuitable for spawning, whereas those at 10L:14D didnot. The results of this study show that limitinglevels for adequate reproductive condition were foundfor some of the factors investigated, however, rangesof the factors above these levels were relativelylarge. That Aphanius dispar's reproductivecondition may be controlled in an artificialenvironment is essential for it to be mass produced,and a prerequisite for its use in the biologicalcontrol of mosquito larvae. 相似文献
Summary Cell free extracts from a streptomycin-resistant E. coli mutant which is also temperature-sensitive for Q phage were studied for suppression of a nonsense mutation at various temperatures. The streptomycin-resistant ribosomes of the mutant were found to be temperature-sensitive in suppression of an amber mutation in f2 phage coat protein while retaining the ability to synthesize proteins at an elevated temperature (42° C). The restriction of amber suppression at 42° C is assumed to be related to an alteration in the ribosomal protein S12 of the streptomycin-resistant mutant which also causes a change in its electrophoretic mobility. 相似文献
Processing of spatio-temporal information in the human visual system has been investigated thoroughly during the past decade, but is still far from being properly understood. Moreover, the theory of separation of information by means of sustained and transient channels already at the retinal level is not satisfactory, as experimental results indicate that these two types of channels span a continuum of temporal characteristics. It is however obvious, that the process of pattern recognition and velocity perception calls for their separation at some level of the hierarchy. In this communication, we extend our model of three-dimensional spatio-temporal frequency expansion in the visual system (Gafni and Zeevi, 1977) to show how velocity-information extraction channels, sensitive to direction and velocity exclusively, can be formed by simple summation of signals from well-defined sets of channels representing points in the frequency space. Correspondence of these channels to characteristics of the cortical neurons is discussed. 相似文献
Astroglia are neural cells, heterogeneous in form and function, which act as supportive elements of the central nervous system; astrocytes contribute to all aspects of neural functions in health and disease. Through their highly ramified processes, astrocytes form close physical contacts with synapses and blood vessels, and are integrated into functional syncytia by gap junctions. Astrocytes interact among themselves and with other cells types (e.g., neurons, microglia, blood vessel cells) by an elaborate repertoire of chemical messengers and receptors; astrocytes also influence neural plasticity and synaptic transmission through maintaining homeostasis of neurotransmitters, K+ buffering, synaptic isolation and control over synaptogenesis and synaptic elimination. Satellite glial cells (SGCs) are the most abundant glial cells in sensory ganglia, and are believed to play major roles in sensory functions, but so far research into SGCs attracted relatively little attention. In this review we compare SGCs to astrocytes with the purpose of using the vast knowledge on astrocytes to explore new aspects of SGCs. We survey the main properties of these two cells types and highlight similarities and differences between them. We conclude that despite the much greater diversity in morphology and signaling mechanisms of astrocytes, there are some parallels between them and SGCs. Both types serve as boundary cells, separating different compartments in the nervous system, but much more needs to be learned on this aspect of SGCs. Astrocytes and SGCs employ chemical messengers and calcium waves for intercellular signaling, but their significance is still poorly understood for both cell types. Both types undergo major changes under pathological conditions, which have a protective function, but an also contribute to disease, and chronic pain in particular. The knowledge obtained on astrocytes is likely to benefit future research on SGCs.
Hydrobiologia - After publication of the above mentioned article, previously undisclosed material was made available resulting in an altered population status for the below mentioned species: 相似文献