SWAP-70 is important for invasive phenotypes of mouse embryo fibroblasts transformed by v-Src

SWAP-70 is a protein involved in actin rearrangement, especially in membrane ruffling. Mouse embryo fibroblasts (MEFs) deficient in SWAP-70 show impaired membrane ruffling and fail to grow in soft agar after transformation by v-Src. Here, we show that v-Src transformed MEFs expressing SWAP-70 are highly invasive. MEFs expressing SWAP-70 or v-Src alone were far less invasive, suggesting that both proteins were required for the cells to be invasive. Expression of both SWAP-70 and v-Src induced constant membrane ruffling, which may cause vigorous cell movement, probably required for invasiveness of the cells. Expression of v-Src alone morphologically transformed MEFs but formed lamellipodia rather than membrane ruffles, suggesting less aggressive nature of the cells compared with those expressing both SWAP-70 and v-Src. These results suggest that v-Src and SWAP-70 act synergistically in the invasion activity of MEFs.     

Urokinase produced by cultured human kidney embryo cells

Urokinase was obtained from cultured cells of human fetal kidneys. Ultrafiltration on an Amicon cell and purification by gel filtration chromatography (Ultrogel ACA54) yielded two molecules capable of activating plasminogen into plasmin. Their molecular weights were respectively 47,500 and 31,500 daltons. The first one showed more active than the latter. In this experiment, only small amounts of Urokinase were harvested. The yield could be enhanced using activators (pronase, glycine) or adapting fetal cells to large scale cultures.     

Transformation of dog embryo kidney cells by human herpesviruses

The infection of dog embryo kidney (DEK) cells with herpes simplex virus type 2 (HSV-2) or human cytomegalovirus (HCMV) led to the development of transformed cell lines. Rapidly dividing DEK cells with unlimited division potential exhibited growth in 2% serum, contained nuclear virus antigens, and formed small (+/- 0.2 mm) colonies in 0.3% agarose. Immortal cell lines showing the same transformation properties were also obtained after transfection with purified HSV-2 or HCMV DNA. These results confirm the transforming capacity of both herpesviruses as well as the usefulness of this different type of mammalian cells in transformation studies.     

Early human embryo metabolism

Non-invasive microanalytical methods have been devised to study the energy metabolism of single human preimplantation embryos. Psyruvate, which is added routinely to all media used to culture human embryos, is consumed throughout the preimplantation period, with glucose assuming an increasing role at embryo compaction and blastocyst formation. All of the glucose consumed may be accounted for by the appearance of lactate in the incubation medium. The enzyme hexokinase my be involved in regulating this aerobic glycolysis. There is cosiderable indirect evidence for the utilisation of endogenous as opposed to exogenous energy substrates, the most likely candidate being protein. Information on early human embryo metabolismis likely to find application in a number of areas: these include the improvement of techniques for assisted human conception, notably in the selection of embryos for transfer following In Vitro Fertilisation; the diagnosis of gentic defects at the preimplantation stage; increased undersding of the causes of implantation failure and miscarriage, and the development of novel post-coital contraceptives.     

Expression dynamics of transforming phenotypes in X-irradiated Syrian golden hamster embryo cells

We investigated the dynamics of expression for morphological transformation, for anchorage-independently growing (Anch-) cells and for mutation at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus in X-irradiated Syrian golden hamster embryo (SHE) cells. No Anch- cells were detected with 0-14 days of post-irradiation incubation before selection. No mutants at the HGPRT locus were detected with 0-5 days of post-irradiation incubation before selection. The maximum number of mutants for all doses was found after post-irradiation incubation for 8 days. On the other hand, the highest frequency of morphological transformants for all doses was detected with 0 days of post-irradiation incubation. The frequency of induction of morphological transformants increased with increasing dose. Then morphological transformants abruptly decreased with increasing lengths of post-irradiation incubation and no morphological transformants were detected with 14 days of post-irradiation incubation before selection (less than 10(-4)). A large fraction of morphological transformants (more than 86%) was cloned with feeder cells and expressed more extensive phenotypes of malignant transformation, such as the acquisition of anchorage-independent growth, immortality in vitro and tumorigenicity during further subculturing.     

耐碳青霉烯类肺炎克雷伯菌血流感染危险因素及预后分析

目的 探讨碳青霉烯类耐药肺炎克雷伯菌血流感染（CSKP）的危险因素以及影响患者28 d预后的相关因素。方法 回顾性分析我院2016年1月至2017年12月期间住院的肺炎克雷伯菌血流感染患者的临床病史资料,按患者血培养标本采集后28 d内预后情况分为存活组与死亡组,应用单因素分析及多因素Logistic回归分析探讨碳青霉烯耐药肺炎克雷伯菌血流感染的危险因素,应用Cox回归分析研究影响肺炎克雷伯菌血流感染28 d预后的相关因素。结果 耐碳青霉烯类肺炎克雷伯菌血流感染的危险因素包括高APACHEⅡ评分、高Pitt菌血症评分、感染时入住ICU、感染前30 d内手术、有创操作、深静脉置管、有创机械通气、器官移植、使用免疫抑制剂、感染前3个月内入住ICU和感染前使用抗菌药物。Logistic回归分析显示高APACHEⅡ评分（OR=1.066,95% CI：1.027~1.107,P=0.001）、手术（OR=3.777,95% CI：1.816~7.855,P<0.001）、有创操作（OR=2.864,95% CI：1.303~6.295,P=0.009）、器官移植（OR=3.892,95% CI：1.553~9.752,P=0.004）、感染前使用抗菌药物（OR=5.626,95% CI：2.740~11.553,P<0.001）是发生碳青霉烯类耐药的肺炎克雷伯菌血流感染的独立危险因素。影响肺炎克雷伯菌血流感染28 d预后的相关因素有高APACHEⅡ评分、高Pitt菌血症评分、感染时入住ICU、感染前30 d内手术、有创操作、深静脉置管、有创机械通气、器官移植、感染前3个月内ICU入住史、使用抗菌药物、粒细胞缺乏、血液透析和菌株对碳青霉烯类耐药。Cox回归分析发现高APACHEⅡ评分（HR=1.061,95% CI：1.039~1.084,P<0.001）、有创操作（HR=2.505,95% CI：1.239~5.063,P=0.011）、入住ICU（HR=1.589,95% CI：1.042~2.424,P=0.031）是影响患者预后的独立危险因素。耐碳青霉烯类肺炎克雷伯菌（CRKP）血流感染患者的28 d病死率明显高于碳青霉烯类敏感肺炎克雷伯菌感染患者（χ2=41.612,P<0.001）。结论 高APACHEⅡ评分、手术、有创操作、器官移植、感染前使用抗菌药物可导致耐碳青霉烯类肺炎克雷伯菌血流感染的发生风险增加。CRKP血流感染患者死亡率显著高于CSKP感染者,但CRKP感染并非患者短期死亡的独立危险因素。而高APACHEⅡ评分、有创操作、入住ICU则可显著增加患者短期病死率。     

Novel phenotypes and developmental arrest in early embryo specific mutants of maize

Embryo specific (emb) mutants exhibit aberrant embryo development without deleterious effects on endosperm development. We have analyzed five emb mutants of maize, which, based on their developmental profiles can be divided into two groups: mutants arrested at early stages and mutants with novel phenotypes. The members of the first group resemble wild-type proembryos and never reach other developmental stages. In the second group the tube-shaped mutants emb*-8522 and emb*-8535 completely lack apical-basal differentiation, while in mutant emb*-8516 a second embryo-like structure arises from the suspensor. The five emb mutations analyzed are non-allelic and two of the mutations are very likely caused by insertion of the transposon mutator, opening the door for their molecular analysis. Received: 10 February 1999 / Accepted: 7 July 1999     

Immunohistochemical study of cell phenotypes in organ culture of the rat embryo liver

The organ culture of the liver of a 14 day old rat embryo was used for immunohistochemical recognition of the available cell types. This culture contains the tissue explant, confluent monolayer cells and a zone of single cells (ZSC). At the start of cultivation a monolayer is formed at the expense of cells slipping down together from the explant, and later these cells were seen to migrate actively from the explant and to proliferate. ZSC is formed by cells migrating from the monolayer. Desmin-containing cells (hepatoblasts, Ito cells and myofibroblasts) migrate from the explant to the monolayer and ZSC. In the monolayer hepatoblasts lose gradually desmin and are converted into cytokeratin 18 (CK-18) expressing hepatocytes. In ZSC, hepatoblasts lose desmin, but no CK-18 synthesis occurs in them. The Ito cells (hypothetical progenitor cells) are spreading over the whole culture, and desmin expression in them does not change. The embryonal Ito cells may transform presumably into myofibrils. Myofibroblasts lie flat on the periphery of ZSC. Besides, desmin myofibroblasts express alpha-actin from smooth muscles. Expression of CK-18 in cells depends on the pattern of intercellular interactions. In the monolayer, CK-18 expression extends successively via the adjacent cells towards the explant. In hepatocytes, migrating into ZSC, CK-18 expression stops. In ZSC, CK-18 expression reappears in compact clusters of stopped cells. It supposed that during formation of close contacts in the monolayer the cells-predecessors may be differentiated into hepatocytes, whereas in the case of disturbance of intercellular connections in ZSC the cells-progenitors may be converted into myofibroblasts. However, on the reconstruction of cell contacts in motionless dense clusters the cells in ZSC are differentiated into hepatocytes.     

Defining bad stroma in human breast tumors

Comment on: Witkiewicz AK, et al. Cell Cycle 2011; 10:1794-809.     

Histopathological prognostic factors of malignant melanoma

The incidence of melanocytic skin lesions, including malignant melanoma has increased in the past few years; histopathologists and dermatopathologists have to face them more often. The correct treatment of melanoma patients by the oncodermatologist and oncologist is based on the histopathological report containing the most important histological prognostic factors. However, the accurate interpretation of these factors may be difficult in the everyday practice, especially in reporting tumor thickness, the level of invasion, the type of exulceration and regression. It is important to standardize the content of the histopathological reports in a reproducible way.     

Production of growth factors by type 5 adenovirus transformed rat embryo cells

Transformation of Sprague-Dawley rat embryo (RE) cells and a cloned Fischer rat embryo cell line (CREF) with wild-type (Ad5) or a temperature-sensitive DNA-minus mutant (H5ts125) of type 5 adenovirus results in a reduction in binding of epidermal growth factor (EGF) to cell surface receptors. A reduction in EGF binding is also seen in a Syrian hamster embryo cell line transformed by a hexon mutant of Ad5. In contrast, a human embryonic kidney cell line (293) transformed by sheared Ad5 DNA or transfected clones of KB cells expressing the E1 transforming region of Ad5 do not show a decrease in receptor binding. When cocultivated, the adenovirus transformed rat cells were able to induce the growth in agar of normal CREF cells. Medium from Ad5 transformed RE cells stimulated the growth in agar of CREF cells and also inhibited [125I]-EGF binding in CREF cells. When fractionated by gel filtration, two peaks of [125I]-EGF inhibiting activities were obtained with apparent molecular weights of 35,000 and 16,000. These results provide the first evidence that cells transformed by an adenovirus can produce a growth factor(s) that inhibits EGF-receptor binding and induces anchorage-independent growth of normal cells.