Detection of radiolabelled bone marrow cells bearing surface ace immunoglobulins by combined autoradiography and immunoperoxidase

A method is described for the simultaneous detection of radiolabelled bone marrow cells bearing surface immunoglobulins by combined autoradiography and immunoperoxidase. Bone marrow cells from normal CBA mice prelabelled in vivo with ¹²⁵IUDR or exposed in vitro to [³H]thymidine were incubated with rabbit anti-mouse immunoglobulins under capping conditions, washed, cytocentrifuged and treated with methanol and hydrogen peroxide to destroy endogenous peroxidase. Cells were then covered with peroxidase-conjugated goat anti-rabbit immunoglobulins, washed, treated with diaminobenzidine a and hydrogen peroxide and finally covered with autoradiographic stripping film and exposed for different times. Peroxidase-positive cells were typically capped and those radiolabelled had autoradiographic silver grains overlying the nucleus.     

Localization of the disulfide bridges of human histocompatibility antigens.

Both papain-solubilized and detergent-solubilized human histocompatibility antigens have been treated with NTCB (2-nitro-5-thiocyanatobenzoic acid) which cleaves these molecules at cysteine residues. A study of the fragment produced has made it possible to deduce the size and location of the two disulfide loops in these molecules. The sizes of the two loops in HLA-B7 and in the mixture HLA-B7 + 12 are about 5100 and 6600 daltons, a size similar to that of the disulfide loops found in immunoglobulins. The disulfide loops in HLA-A2 may be smaller in size. The two loops are located in middle regions of these molecules; neither the N-terminal nor the C-terminal regions contain disulfide loops.     

Effect of o,p-dichlorodiphenyldichloroethane and perthane in vitro on glutathione reductase activity in the adrenals of dogs and guinea pigs

o,p-Dichlorodiphenyldichloroethane (o,p-DDD) and Perthane, when added in a concentration of 312 M to homogenate and cytoplasmic fraction of dog adrenals, activate glutathione reductase. In a concentration of 156 M, o,p-DDD and Perthane do not affect glutathione reductase activity of the dog adrenals. When given in vitro, o,p-DDD and Perthane activate glutathione reductase of the guinea pig adrenals. o,p-DDD has no effect on glutathione reductase activity of the cytoplasmic fraction of dog liver and kidney, thus confirming the high specificity of its effect on the adrenal cortex.Laboratory of Pathophysiology, Institute of Endocrinology and Metabolism, Kiev. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 2, pp. 159–161, February, 1978.     

Fluoropyrimidine chemotherapy in a rat model: comparison of fluorouracil metabolite profiles determined by high-field 19F-NMR spectroscopy of tissues ex vivo with therapy response and toxicity for locoregional vs systemic infusion protocols

A Sprague-Dawley rat model with DS sarcoma transplanted in the thigh was used to compare transcatheter locoregional i.a. and systemic i.v. administration of 5-fluorouracil (FU) at 12 dose-rate schedules: 25, 50 and 100 mg/kg; bolus, 1, 5 and 24 h infusions. In experiment A tumor (62/67 animals) as well as liver and kidney (56/67 animals) were excised 1 h after a single bolus or 1 h infusion or at the end of 5 and 24 h infusions. (19)F-NMR spectroscopy at 11.7 T was used to quantitate FU and its metabolites in ca. 1 g of tissue at 4 degrees C. In experiment B analogous FU treatments were repeated for 5 days (rats 80+11 controls). Tumor volumes vs time, various blood parameters and survival times were recorded, and a log growth rate parameter log GR, a response index RI, and a toxicity index TI were calculated. The i.a. vs i.v. ratios for tumor concentrations of FU and total anabolites (F-Nucl) were >1 for nearly all treatments and increased with infusion time at the higher doses. F-Nucl in tumor correlated linearly with total fluorine concentration (Tot. F range 30-1100 nmol/g) over all treatments (r=0.92, slope=0.45, p<0.0001). For non-bolus i.v. treatments [FU+F-Nucl] decreased linearly with decreasing FU dose rate (r(2)=0.74, zero intercept), while i.a. treatments showed non-linear behavior. For non-bolus treatments the mean log GR per treatment group showed a negative correlation (r=-0.87) with log[F-Nucl]. The most effective non-toxic treatments were 25 mg/kg over 5 or 24 h; the i.a. route was superior to i.v. on the basis of [FU+F-Nucl], RI, the reduction in log GR, and Kaplan-Meier survival statistics. For liver and kidney Tot. F (>83% FU catabolites) reached ca. 3-4 and 6-7 micromol/g, respectively, at the highest dose rates for either route; F-Nucl were detected only for Tot. F>500 nmol/g and increased exponentially as Tot. F increased (toxic treatments). The concentrations of the main catabolite (alpha-fluoro-beta-alanine, FBAL) in tumor did not correlate with Tot. F but rather with FBAL levels in kidney (r=0.90, all treatments), indicating that uptake of liver-derived FBAL from the circulation is the major source of FBAL in tumor.     

Extrapleural solitary fibrous tumor: clinicopathologic study of 17 cases and molecular analysis of the p53 pathway

Solitary fibrous tumor (SFT) occurring at various extrapleural sites is sometimes difficult to diagnose because of its histologic variability. Although a solitary fibrous tumor is usually a slow-growing tumor with favorable prognosis, a small number of malignant cases have been reported. In the present study, we examined the clinical behavior, histologic, immunohistochemical and molecular features of 17 cases of extrapleural SFT. Four tumors were located in the pelvic cavity, two in the nasal cavity, two were confined to the pulmonary parenchyma, and there was one each in the meninges, kidney, mediastinum, retroperitoneum, temporal region, neck, groin, buttock and thigh. Histologically, all the tumors were characterized by the presence of areas consisting of a proliferation of bland spindle cells with variable amounts of thick, often hyalinized or keloid-like intercellular collagen bundles. Highly cellular areas were observed in three tumors, frequent mitoses in two, and cellular pleomorphism and tumor necrosis in one each. All 17 tumors showed immunoreactivity to CD34 and 15 (88%) to bcl-2 protein. The labeling indices of p53, mdm2 protein and Ki-67 were generally low. PCR-SSCP and a subsequent sequence analysis of the p53 gene disclosed point mutation at codon 161 in exon 5 in one of the 13 cases analyzed. According to follow-up information, none of the patients had developed local recurrence or distant metastasis. Our results suggest that most extrapleural SFTs behave in a benign fashion even in a higher histologic grade group, and it is difficult to predict their clinical outcome. Complete surgical excision in order to obtain clear margins and long-term follow-up is advisable for patients with an extrapleural SFT.     

干扰素a-2b对增生性瘢痕HLA-DR和CD1a分子的影响

目的:探讨干扰素a-2b对增生性瘢痕HLA-DR和CD1α。分子的作用。方法 利用免疫组织化学方法检测6例增生性瘢痕（HS）和6例正常皮肤HLA-DR和CD1α分子的分布及密度,观察干扰素a-2b治疗后HS HLA-DR和CD1α分子的变化。结果:（1）HS组织表皮HLA-DR LC的数量806.67±101.72个//mm2和表皮CD1α LC的数量700.00±108.82个/mm2明显高于正常皮肤510.00±45.07个/mm2,521.24±57.87个/mm（P<0.05）。（2）HS组织HLA-DR分子在角质形成细胞和成纤维细胞中异常出现。（3）于扰素a-2b治疗3d时HS表皮HLA-DR LC及CD1α LC的数量分别为283.34±73.12个/mm2,220.00±83.92个/mm2,7d时516±49.67个/mm2,589.10±108.82个/mm2,干扰素a-2b治疗后HS表皮HLA-DR LC及CD1αLC的数量明显降低（P<0.05）。结论:（1）HLA-DR和CD1α分子数量的增加提示HS和K局部组织可能处于高兔疫应答状态;（2）干扰素a-2b可能通过抑制HLA-DR和CD1α分子降低HS高免疫应答的状态。     

Sequence analysis of exons 1, 2, 3, 4 and 5 of the HLA-B5/35 cross-reacting group

The HLA-B5/35 cross-reacting group (CREG) is a set of closely related antigens including HLA-B35, B51, B52, B53 and B78. The nucleotide sequences of exon 1 through 5 of the B5/35 CREG were determined to assess the level of polymorphism. For exons 2 and 3, the previously described sequence-based typing (SBT) strategy was applied, the nucleotide sequences of exon 1, 4 and 5 were determined by allele-specific sequencing. A total of 225 unrelated individuals were HLA-B typed by heterozygous sequencing of exons 2 and 3. In the B5/35 CREG, 26 different alleles were identified, whereas 63 non-B5/35 CREG alleles were sequenced. The SBT strategy was proven to be reliable and efficient for high resolution typing of the B5/35 CREG. The nucleotide sequences of exon 1, 4 and 5 were determined for the 26 different B5/35 CREG alleles to establish the level of polymorphism. For seven different alleles, of which the exon 1, 4 and 5 sequences were hitherto unknown, the sequences were elucidated and in agreement with the known B5/35 sequences. Nineteen HLA-B5/35 CREG alleles with previously published exon 1, 4 and 5 sequences were sequenced in at least two individuals. Three new alleles were identified. The first, B*5204, showed a difference at position 200 compared to B*52011, which was previously considered a conserved position. The other two alleles, B*3542 and B*51015, showed exon 2 and 3 sequences identical to B*35011 and B*51011, but differences in exons 1 and 4, respectively. B*3542 had differences at position 25 and 72 and B*51015 showed a difference at position 636. More polymorphism might be present outside exons 2 and 3 than previously thought.     

Mapping of sporulation-specific functions in the yeast syntaxin gene<Emphasis Type="Italic"> SSO1</Emphasis>

The yeast Saccharomyces cerevisiae has two closely related plasma membrane syntaxins, Sso1p and Sso2p, which together provide an essential function in vegetative cells. However, Sso1p is also specifically needed during sporulation; and this function cannot be provided by Sso2p. We used fusions between SSO1 and SSO2 to map the sporulation-specific function of SSO1. We found that the two N-terminal -helices Ha and Hb of Sso1p are important for sporulation, since it is reduced 8-fold for fusions where Ha and Hb are derived from Sso2p. In contrast, the C-terminal half of Sso1p does not seem to be specifically required for sporulation. Surprisingly, we further found that the 3 untranslated region (3UTR) of SSO1 is essential for sporulation. Western blots failed to reveal a preferential expression of Sso1p in sporulating cells, indicating that effects on gene expression are unlikely to explain why the SSO1 3UTR is needed for sporulation.Communicated by S. Hohmann     

定向诱导骨髓间充质干细胞向心肌细胞分化

目的:采用骨髓间充质干细胞体外转化为心肌样细胞,为心衰的干细胞移植治疗提供基础。方法:分离培养大鼠骨髓间充质干细胞,5-氮杂胞苷诱导24h后正常培养,通过形态学、PAS反应、磷钨酸苏木素染色(PTAH)、免疫细胞化学染色作鉴定。结果:诱导细胞以梭形、柱状为主,部分细胞有分支,单个核、卵圆形、居中,似心肌细胞,PAS及PTAH染色均为阳性。诱导后培养2周的细胞表达Sarcomeric Actin和Connexin-43较弱,以后表达逐渐增强,而且1、3、5代细胞均稳定表达。结论:骨髓MSCs可在体外被诱导分化为心肌样细胞,有望成为心衰干细胞移植治疗的理想细胞材料。     

经舟骨月骨周围脱位的诊断和治疗

目的 探讨经舟骨月骨周围脱位的诊断和治疗。方法 回顾性分析17例经舟骨月骨周围脱位病例，背侧型13例，掌侧型4例；新鲜损伤组10例中8例行撬拨复位成功，仅2例因舟骨对位不良接受手术治疗；陈旧性组7例中均接受手术，4例行开放复位内固定，3例行近排腕骨切除术。结果 采用Cooney评价标准，优11例，良3例，可1例，差2例，优良率82．35％。结论 经舟骨月骨周围脱位的早期治疗简单且疗效较好，而一旦误诊，则多数需手术治疗，且残留不同程度的功能障碍。提高对本病损伤机制、腕部体征和影像学特点的认识和了解，减少误诊。是提高本病预后的关键因素之一。