Ultrastructural immunostaining of infiltrating ductal breast carcinomas with the monoclonal antibody H: a comparative study with cytokeratin 8

The monoclonal antibody H (mAbH) detects an epitope consisting of an O-linked N -acetyl glucosamine (O-GlcNAc) and neighboring amino acids. This epitope has been found by using extracts from the MCF-7 human breast carcinoma cell line in immunoblotting experiments, on cytokeratin 8 (CK8) and 5 other polypeptides. In the present study, a double immunogold method was applied for the colocalization of CK8 and mAbH epitope on epoxy thin sections in 18 cases of infiltrating ductal breast carcinomas (IDBC) and in 6 cases of fibroadenomas, to study the accurate subcellular distribution of CK8 in breast cancer cells, as compared to the 5 polypeptides, recognized by mAbH. Furthermore, a detailed quantitative evaluation of the double immunolocalization over the cellular compartments of cancer cells was undertaken with the aid of a computerized image analysis system and the results were assessed statistically. The distribution pattern of CK8 and the mAbH epitope in the neoplastic mammary epithelial cells was similar in IDBC as compared to fibroadenomas, while the gold labeling intensity of these epitopes differed over the cellular compartments between malignant and benign biopsies. The results reveal the significance of the role of CK8 and O-GlcNAc glycosylation in the biology of the neoplastic mammary cells in vivo, determining their malignant potential.     

Bone remodeling analysis of various dental implant surfaces using polyfluorochrome sequential labeling in rabbit tibias

The purpose of this study is to analyze the bone remodeling process after the placement of threaded implants with rough (RBM) and hydroxyapatite coated surfaces (HA) in rabbit tibias using polyfluorochrome sequential labeling. Histomorphometry was performed in order to quantify the amount of each label deposited during the healing period. This work demonstrates the possibility of periodic identification of apatite deposition during bone remodeling around titanium and ceramic implants. It has been concluded that the polyfluorochrome sequential labeling is an important tool for identification of bone remodeling after the insertion of titanium and ceramic implants inside rabbit tibias.     

The proliferative pattern of the prostatic carcinoma before and under hormonal treatment

Summary 387 carcinomas of the prostate gland were examined histologically and partly by autoradiography. 45.7% of the carcinomas were uniformly structured whereas 54.3% showed a pluriform pattern. Highly and poorly differentiated adenocarcinomas predominated. Very often cribriform structures were found besides other carcinomatous parts.Autoradiographically the labeling indices of adenocarcinomas were extremely low (0.69% and 0.83% respectively). Higher values could be observed in cribriform and solid anaplastic carcinomas with labeling indices up to 7.0%. The mitotic indices of adenocarcinomas and anaplastic carcinomas ranged between 0.01–0.6%. In the hyperplastic glandular epithelia in the vicinity of carcinomas a labeling index was found which was twice as high as in nodular hyperplasia without carcinoma.Adenocarcinomas treated with estrogens showed decreased labeling indices. Changes in the labeling indices were not observed in one case of a adenocarcinoma and in two cases of cribriform and solid anaplastic carcinomas. These results are in good agreement with those described in the literature, in which it is pointed out, that glandular carcinomas have a better sensibility to hormonal therapy than solid anaplastic or cribriform carcinomas of the prostate.Supported by Landesamt für Forschung, Minister für Wissenschaft und Forschung des Landes Nordrhein-Westfalen, Düsseldorf     

Perceptions and Attitudes Toward Prenatal Cocaine Exposure in Young Children

Objective: This paper investigated perceptions and attitudes toward young children believed to be prenatally exposed to cocaine. Method: 37 college students watched a videotape of eight children doing the same task and rated each child's performance after being told if the child had, might have been, or had not been prenatally exposed to cocaine. Results: Multiple analysis of variance confirmed that when a child was labeled prenatally exposed to cocaine, the participants generally scored the child lower and less positively than if the child was thought to be possibly exposed or not exposed at all. Ethnicity and gender differences were also found. Participants rated males higher than females and African-American children higher than Caucasian children. Conclusions: Individuals who believe a child is exposed prenatally to cocaine hold more negative perceptions regarding that child's developmental abilities. These negative perceptions of a child labeled prenatally exposed to cocaine may diminish expectations for that child's developmental potential.     

Radiobiologic significance of apoptosis and micronucleation in quiescent cells within solid tumors following γ-ray irradiation

Purpose: To determine the frequency of apoptosis in quiescent (Q) cells within solid tumors following γ-ray irradiation, using four different tumor cell lines. In addition, to assess the significance of detecting apoptosis in these cell lines.

Methods and Materials: C3H/He mice bearing SCC VII or FM3A tumors, Balb/c mice bearing EMT6/KU tumors, and C57BL mice bearing EL4 tumors received 5-bromo-2′-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received γ-ray irradiation at a dose of 4–25 Gy while alive or after tumor clamping. Immediately after irradiation, the tumors were excised, minced, and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling (= Q cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 hours after irradiation, tumor cell suspensions obtained in the same manner were fixed. The apoptosis frequency in Q cells was also determined with immunofluorescence staining for BrdU. The MN and apoptosis frequency in total (P + Q) tumor cells were determined from the tumors that were not pretreated with BrdU.

Results: In total cells, SCC VII, FM3A, and EMT6/KU cells showed reasonable relationships between MN frequency and surviving fraction (SF). However, fewer micronuclei were induced in EL4 cells than the other cell lines. In contrast, a comparatively close relationship between apoptosis frequency and SF was found in total cells of EL4 cell line. Less apoptosis was observed in the other cell lines. Quiescent tumor cells exhibited significantly lower values of MN and apoptosis frequency probably due to their large hypoxic fraction, similar to total tumor cells on clamped irradiation.

Conclusion: γ-ray irradiation induced MN formation in SCC VII, FM3A, and EMT6/KU tumor cells, and the apoptosis was marked in EL4 cells compared with the other cell lines. Our method for detecting the Q cell response to γ-ray irradiation using P cell labeling with BrdU and the MN frequency assay was also applicable to apoptosis detection assay.     

Two routes to [11C-carbonyl]organo-isocyanates utilizing [11C]phosgene ([11C]organo-isocyanates from [11C]phosgene).

Two generic radiosynthetic routes for the preparation of [¹¹C-carbonyl]isocyanates have been developed. Reaction of N-organo-sulfinylamines; RNSO, (R = Me, Et, allyl, cyclohexyl and phenyl) with [¹¹C]phosgene gave the corresponding [¹¹C-carbonyl]isocyanates in good radiochemical yield (53–68%) from [¹¹C]phosgene (decay corrected) in ca 16 min from EOB. Alternatively, the reaction of [¹¹C]phosgene with N,N′-organo-ureas; (RNH)₂CO, (R = Me, Et, Pr and phenyl) also gave the corresponding [¹¹C-carbonyl]isocyanates in moderate radiochemical yield (9–37%) from [¹¹C]phosgene (decay corrected) in ca 16 min from EOB. For identification, the [¹¹C-carbonyl]organo-isocyanates were derivatized with 1-(2-methoxyphenyl)piperazine in situ to [¹¹C-carbonyl]carboxamides and the position of radiolabelling in the carbonyl group confirmed by [^11/13C]co-labeling and subsequent carbon-13 NMR spectroscopy.     

APAAP免疫酶标及DNA末端原位标记双重染色检测急性白血病细胞原位凋亡

目的 :观察原位细胞凋亡与急性白血病 (AL )发生与转归的关系。方法 :用碱性磷酸酶抗碱性磷酸酶(APAAP)免疫酶标和 DNA末端原位标记 (ISEL )双重染色方法检测 49例急性白血病骨髓涂片中细胞凋亡状况 ,初治组 2 3例与 15例缺铁性贫血病例对照并与 2 6例经治病例对照。结果 :未经治疗的 AL 平均凋亡指数 (AI)明显低于对照组 (P <0 .0 1)。化疗后凋亡明显增加 ,与化疗前比较差异有极显著性意义 (P <0 .0 1)。AI>5 %者平均原始细胞下降指数 (MBDI)为 2 8.6 8% ,AI<5 %者平均 MBDI仅为 4.9%。凋亡细胞大多数为白血病细胞。非白血病细胞凋亡亦有增加。结论 :AL 发生与凋亡逃逸有关 ,细胞毒化疗的主要机理之一为促进细胞凋亡。化疗后全血细胞下降与正常造血细胞凋亡增加有关。     

猪骨髓间充质干细胞超顺磁性氧化铁标记方法的实验研究

目的：探索超顺磁性氧化铁纳米颗粒（SPIO）标记猪骨髓间充质干细胞（MSCs）的合适方案。方法：分离培养猪MSCs,用不同SPIO浓度、不同共孵育时间进行标记,根据标记效率指标（普鲁士蓝染色、细胞铁浓度测定、细胞透射电镜检查）和标记毒性指标（台盼蓝排除试验、CCK-8试验）确定适宜的标记条件,然后对不同标记细胞进行磁共振（MR）体外成像。结果：（1）随着SPIO浓度的增加,MSCs细胞内平均含铁量相应升高;细胞涂片普鲁士蓝染色也显示细胞内蓝染颗粒相应增加。当SPIO浓度为25、50μg·mL-1时,细胞存活率和细胞活力与无SPIO时差异无统计学意义（P〉0.05）;但当SPIO浓度增加到100μg·mL-1时,CCK-8试验显示细胞活力下降（P〈0.01）;当SPIO浓度增加到200μg·mL-1时,细胞存活率和细胞活力均下降（P〈0.05）;（2）设定SPIO浓度为50μg.mL-1,随共孵育时间增加,细胞内含铁量逐渐升高（各组间比较均P〈0.01）;当标记时间为0、2、4、8、16、24h时,细胞存活率和细胞活力并差异无统计学意义（P〉0.05）;但当标记时间进一步延长到48h时,细胞存活率有所降低（P〈0.05）;（3）按＂50μg/ml×24h方案＂标记后,1.5TMR成像仪体外MRT2＊ WIflash检测阈值为7.5×104个细胞。结论：50μg·mL-1×24h方案是猪骨髓间充质干细胞比较合适的磁探针标记方案。     

Decision trees for evaluating skin and respiratory sensitizing potential of chemicals in accordance with European regulations

Guidance for determining the sensitizing potential of chemicals is available in EC Regulation No. 1272/2008 Classification, Labeling, and Packaging of Substances; REACH guidance from the European Chemicals Agency; and the United Nations Globally Harmonized System (GHS). We created decision trees for evaluating potential skin and respiratory sensitizers. Our approach (1) brings all the regulatory information into one brief document, providing a step-by-step method to evaluate evidence that individual chemicals or mixtures have sensitizing potential; (2) provides an efficient, uniform approach that promotes consistency when evaluations are done by different reviewers; (3) provides a standard way to convey the rationale and information used to classify chemicals. We applied this approach to more than 50 chemicals distributed among 11 evaluators with varying expertise. Evaluators found the decision trees easy to use and recipients (product stewards) of the analyses found that the resulting documentation was consistent across users and met their regulatory needs. Our approach allows for transparency, process management (e.g., documentation, change management, version control), as well as consistency in chemical hazard assessment for REACH, EC Regulation No. 1272/2008 Classification, Labeling, and Packaging of Substances and the GHS.