A gene transfer strategy for making bone marrow cells resistant to trimetrexate

Trimetrexate (TMTX) is an anticancer drug with potential advantages over the more commonly used antifolate, methotrexate (MTX); however, its use has been limited by severe myelosuppression. Retroviral vectors containing mutant dihydrofolate reductase (DHFR) genes have been used to protect bone marrow cells from MTX, suggesting a similar approach could be used for TMTX. We first screened six variants of human DHFR to determine which allowed maximal TMTX resistance in fibroblasts. A variant enzyme containing a Leu-to-Tyr mutation in the 22nd codon (L22Y) was best, allowing a 100-fold increase in resistance over controls. Murine hematopoietic progenitor cells transduced with an L22Y- containing retroviral vector also showed high-level TMTX resistance in vitro. Mice reconstituted with L22Y-transduced bone marrow cells were challenged with a 5-day course of TMTX to determine whether hematopoiesis could be protected in vivo. Transfer of the L22Y vector resulted in consistent protection from TMTX-induced neutropenia and reticulocytopenia at levels that correlated with the proviral copy number in circulating leukocytes. We conclude that the L22Y vector is highly effective in protecting hematopoiesis from TMTX toxicity and may provide a means for increasing the therapeutic utility of TMTX in certain cancers.     

Immunohistochemically Determined Estrogen and Progesterone Receptor Levels: A Comparison of Three Antibodies with the Ligand-Binding Assay

Abstract: Traditionally, estrogen and progesterone receptor levels have been determined by biochemical ligandbinding assays, but more recently immunohistochemical techniques have become available. They have gained popularity due to their low cost, smaller sample size requirements, and direct visualization capability of reaction location. Several antibody clones are commercially available and antibodies directed against the estrogen receptor (ER) are supplied by Ventana Medical Systems (Tucson, AZ), Abbott Laboratories (Abbott Park, IL), and lmmunotech Westbrook, ME). Antibodies directed against the progesterone receptor (PgR) are supplied by Ventana Medical Systems (Tucson, AZ), lmmunotech (Westbrook, ME), and Becton-DickinsonKelI Analysis Systems (San Jose, CA). Computer-assisted image analysis using the CAS ZOOTM (Becton-DickinsonKIS, San Jose, CA) allows quantitation of immunohistochemically determined receptor levels. Correlation of quantitated immunohistochemical ER levels with values determined by ligand-binding assay revealed the Ventana antibody to most closely predict the ligand-binding results (wk = .667). The Ventana anti-progesterone antibody quantitation most closely correlated with the ligand-binding results (wk = 435) for determination of PgR. Progesterone receptor level as determined by any of the tested methods did not stratify patients into favorable and unfavorable prognostic groups. Estrogen receptor level as determined by the Ventana antibody was the most predictive of patient outcome but this relationship did not reach statistical significance (p = .09). Most discrepancies between the ligand-binding assay and the immunohistochemical assays were associated with one of three factors: (a) low volume of neoplastic cells present due either to small sample size or high stromal content, (b) premenopausal status with circulating endogenous estrogens potentially occupying receptor sites, (c) presence of benign breast epithelium resulting in a false-positive ligand-binding assay.     

吸收度线性组合分光光度法的初步探讨

本文在联立方程法(包括双波长法、系数倍率法)、三波长法、正交函数法的基础上提出一种新的计算分光光度测定法——吸收度线性组合法。文中对上述诸法进行了比较,并从理论和实验上证明了新法只要合理组合,随着测试点数的增加,测定精度可望改善。     

Determination of Proliferation Index By MIB-1 Immunostaining in Early Stage Breast Cancer Using Quantitative Image Analysis

Abstract: Several clinicopathologic variables influence prognosis in breast cancer, including stage, histologic grade, nodal status, and tumor size. Multiple studies have shown an independent value of proliferation index as a prognostic variable for the stratification into favorable and unfavorable groups. The monoclonal antibody MIB-1 reacts with the same antigen site, not epitope, as recognized by the Ki-67 antibody. Like Ki-67, MIB-1 reacts with cells in the late G₁, S, M and G₂ phases of the cell cycle, but MIB-1 has the advantage of reacting with formalin-fixed, paraffin-embedded material. The authors investigated the feasibility of using image analysis to quantitate the MIB-1 antibody staining (proliferation index [PI]) and predict survival in a series of 230 patients with stage I and stage II breast cancer. In a univariate Cox regression model, larger values of MIB-1 were related to shorter survival times (p < 0.001). Exploratory statistical procedures were used to categorize the patients into good, intermediate, and poor survival groups using the following proliferation indices as cut-points: <5%, 5–11%, and >11 %, respectively. Higher clinical stage was associated with higher MIB-1 values and shorter survival (p = 0.01, and p = 0.003, respectively). Tumor size (p = 0.02) and nodal status (p = 0.05) were also associated with higher values of MIB-1. After adjusting for age, clinical stage, nodal status, and tumor size in a multivariate analysis, MIB-1 retained its prognostic significance (p < 0.0001) when considered as either a continuous or categorical variable. There were no significant associations between MIB-1 determined proliferation index and age (p = 0.54), histologic grade (p = 0.69), nuclear grade (p = 0.06) or the presence of vascular invasion (p =.66). There is a strong statistical relationship between cell proliferative activity, as determined by MIB-1 expression, and survival in early stage breast cancer.     

槲寄生化学成分的研究——Ⅷ.2,3-丁二醇单葡萄糖甙的分离和结构

From the ethanol extract of Viscum coloratum (Kom.) Nakai , a giucoside ofaliphatic diol and three other glucosides were isolated. Based on chemical and spectroscopic analysis,the structures have been elucidated as 2-β-D- glucosyl-3- methylpropanol (Ⅷ), syringin (Ⅸ),eleatheroside E(Ⅹ) and syringenin-4'-O-D-apiosylglucoside (Ⅺ). Ⅷ is a new glucoside of aliphaticdiol and named 3-β-D-glucopyranosyloxy-butanol-2. Three other compounds (Ⅸ～-Ⅺ) were foundfor the first time in this plant.     

Inter- and intrasensory modality matching in children with hand-eye coordination problems: exploring the developmental lag hypothesis

This study set out to explore the suggestion that the problems experienced by 8-year-old children diagnosed as clumsy in the area of hand-eye coordination (HECP) might be attributed to a developmental lag. The performances of this group of HECP children were compared with those of groups of 5-year-old and 8-year-old controls without such deficits, when required to carry out a task involving pointing, without vision, to targets located, visually, visually/proprioceptively, or proprioceptively, the dependent variable being the distance error score from the centre of the target. The performances of the HECP children, when vision or vision/proprioception was used to locate the targets, were shown to be inferior to those of the two control groups of children thereby supporting a visual deficit hypothesis. When the targets had to be located proprioceptively, the performance of the HECP children was shown to be similar to that of the 5-year-olds, while both groups were inferior to the 8-year-olds, thereby supporting a developmental lag hypothesis in proprioceptive terms. However, when the scores for the preferred and non-preferred hands were analysed separately a marked deterioration in the performances of both the 5-year-old controls and the HECP children was observed while the 8Jyear-old controls were unaffected. While this finding supports a developmental lag explanation of the inferior performances of the HECP children, it was necessary to qualify such an explanation when the within-group performances using the preferred and non-preferred hands were compared. Only the HECP children, under the visual/proprioceptive or proprioceptive conditions, showed significant performance differences, in. favour of the preferred hand. This finding was taken as a suggestion that the developmental lag exhibited by the HECP children might have pathological overtones possibly related to the development of the corpus callosum.     

槲寄生化学成分的研究——Ⅳ.槲寄生新甙Ⅳ的结构

前文曾报道从槲寄生中分离及确定五个新黄酮甙的结构。本文报道另一新黄酮化合物(Ⅳ)的分离和结构测定。槲寄生(Viscum coloratura(Kom)Nakai)生药的乙醇提取物,分别用石油醚、醋酸乙酯和正丁醇萃取,正丁醇萃取物经聚酰胺细粉和硅胶低压柱层析,分离得到Ⅳ。通过光谱分析和水解试验,确定Ⅳ的结构为鼠李秦素-3-0-β-D-(6″-β-羟基-β-甲基戊二酸半酯)-葡     

The preparation of leukocyte-poor red blood cells: a comparative study

Thirteen methods of prepararing leukocyte-poor red blood cells were compared for their effectiveness in removing leukocytes. When the starting material was six to ten days old, the residual leukocyte count was three to four times lower than when fresh material was used and no granulocytes could be found. The most effective manual procedure was a single upright sedimentation of six- to ten-day-old saline-diluted red cells. Inverted sedimentation was no more effective in leukocyte depletion than upright sedimentation. Automated cell processors were no more effective than several of the better manual methods. Freezing and deglycerolizing resulted in a ten-fold improvement in leukocyte removal with less red blood cell loss but at higher cost than the nonfreezing methods. The study indicates that unless the minimal leukocyte count of frozen red blood cells is essential, a single upright sedimentation of saline-diluted, six- to ten-day-old red blood cells is currently the most effective procedure for preparing leukocyte-poor red blood cells.     

槲寄生化学成分的研究 Ⅶ.槲寄生新甙Ⅶ的分离和结构

自中药槲寄生中分离得到一个新的黄酮甙,经波谱测定及水解试验,确定其结构为鼠李秦素-3-O-β-D-芹糖基(1→2)-[6″-(3-羟基-3-甲基戊二酸半酯)]葡萄糖甙,命名为槲寄生新甙Ⅶ(viscumneoside Ⅶ).