Genetic convergence during serial in vitro passage of a polyclonal squamous cell carcinoma

A cell line was established from an in situ squamous cell carcinoma of the skin (Bowen's disease), and its in vitro karyotypic evolution was cytogenetically analyzed. Initially, considerable genetic heterogeneity was evident. Nine cytogenetically abnormal clones, eight of which were apparently unrelated, were found among the 83 metaphases analyzed from the primary culture and the first passage. With increasing time in culture this complexity was reduced, so that a single clone dominated passages 7-11. The clone that emerged from this genetic convergence had a t(12;17)(p13;q21) as the sole abnormality. Our findings indicate that the cytogenetic multiclonality that has been repeatedly detected in short-term cultures of squamous cell carcinomas is not caused by the in vitro conditions. Instead, the principles of Darwinian selection apply: the altered, but stable, selection pressure facing a newly established and initially multiclonal cell line will lead to a reduction of genetic heterogeneity until the one clone that now has the proliferative advantage outgrows the other subpopulations.     

眼镜蛇科蛇毒对S180,EAC腹水癌治疗作用研究

本文报道金环蛇、扁颈蛇、眼镜蛇、银环蛇蛇毒及其细胞毒对小鼠S180,EAC腹水癌的治疗作用。结果表明,蛇毒及其细胞毒在体外有明显杀灭癌细胞作用,体内有较明显的治疗作用,动物存活时间延长,接种率降低。肿瘤细胞呈现不同程度形态变化,主要为膜破裂,坏死等。治疗效应由强到弱为金环蛇毒,扁颈蛇毒,金环蛇细胞毒,眼镜蛇毒,眼镜蛇细胞毒。银环蛇毒无作用。     

The primary structure of human apolipoprotein A-IV

Human apolipoprotein (apo) A-IV was purified from chylous ascites fluid. Proteolytic peptides produced by trypsin and Staphylococcus aureus V8 proteinase digestions were purified by high-performance liquid chromatography and sequenced. Human apoA-IV contains 376 amino acid residues. The peptide-derived sequence generally matches two previously reported DNA-derived amino acid sequences except for discrepancies in five positions. In order to examine these discrepancies further, one complete apoA-IV cDNA clone and another partial clone were sequenced. Comparison of all the available information indicates that the peptide-derived sequence reported here is accurate. Sequencing errors probably account for some of the discrepancies between the two primary sequences predicted by earlier nucleotide analyses. In certain positions, however, bona fide sequence heterogeneity or cloning artifact cannot be excluded.     

How does l-glutamate transport relate to selection of mixed nitrogen sources in Rhizobium leguminosarum biovar trifolii MNF1000 and cowpea Rhizobium MNF2030?

When growing on a mixture of ammonia and l-glutamate as nitrogen sources, Rhizobium leguminosarum biovar trifolii MNF1000 utilizes ammonia exclusively, while cowpea Rhizobium MNF2030 utilizes both compounds at similar rates. l-Glutamate transport in both strain MNF1000 and MNF2030 is active, giving rise to a 60-fold concentration gradient across the membrane of cells of strain MNF2030. Both strains produce two kinetically distinguishable glutamate transport systems under all conditions of growth — a high affinity system with an apparent K _m of 0.06–0.17 M but of relatively low V _max, and a low affinity system with a K _m of 1.2–6.7\ M, but of higher overall capacity. l-Glutamate transport activity in cells of MNF2030 was relatively insensitive to the presence of ammonia in the growth medium. By contrast, ammonia in the growth medium resulted in low activities of glutamate transport in cells of MNF1000 which were provided with a carbon source, offering one explanation for the failure of this strain to use glutamate in the presence of ammonia. However, in cells of MNF1000 growing on glutamate as sole source of carbon and nitrogen, the glutamate transport system is synthesized, even in the presence of accumulated or added ammonia. This suggests that the regulation of the glutamate permease also depends on availability of carbon source.Abbreviations CCCP carbonyl cyanide m-chlorophenyl hydrazone - HEPES N-hydroxyethylpiperazine-N-2-ethanesulphonic acid     

4-aminobutyrate is not available to bacteroids of cowpea Rhizobium MNF2030 in snake bean nodules

Laboratory cultures of cowpea Rhizobium MNF2030 grew on 4-aminobutyrate (GABA) as sole source of carbon and nitrogen. GABA transport was active since it was inhibited by carbonyl cyanide mchlorophenyl hydrazone and 2,4-dinitrophenol and cells developed a 400-fold concentration gradient across the cell membrane. Arsenite treatment of GABA-grown cells revealed stoichiometric conversion of GABA to pyruvate, indicating that 2-oxoglutarate is not an intermediate in GABA catabolism. GABA catabolism by cells of strain MNF2030 grown on GABA appreared to involve GABA transaminase, succinic semialdehyde dehydrogenase and malic enzyme; the first two enzymes were specifically induced by growth on GABA. The deamination process and removal of NH₃ in cells catabolizing GABA involved GABA: 2-oxoglutarate transaminase; glutamate: oxaloacetate aminotransferase; asparate: pyruvate aminotransferase and alanine dehydrogenase.Isolated snakebean bacteroids of strain MNF2030 transported only small amounts of GABA and had uninduced levels of GABA catabolic enzymes, even though the nodules contained significant levels of GABA. The data suggest that GABA is not available to snakebean nodule bacteroids, presumably because of a control imposed by the peribacteroid membrane.Abbreviations CCCP Carbonyl cyanide m-chlorophenyl hydrazone - HEPES N-hydroxyethylpiperazine-N-2-ethanesulphonic acid - DTT dithiothreitol - SSAD succinic semialdehyde dehydrogenase - GABAT 4-aminobutyrate transaminase - GABA 4-aminobutyrate     

Interactions of chiral molecules with an (r)-n-(3,5-dinitrobenzoyl) phenylglycine HPLC stationary phase

Forty different chiral molecules were studied by liquid chromatography with a Pirkle-type, (R)-N-(3,5-dinitrobenzoyl) phenylglycine (DNBPG), chiral stationary phase column. The dramatic effect of a small molecular change on chiral recognition was demonstrated using DL-amino acid derivatives. The inductive effect on chiral recognition was also studied using trifluoro-, trichloro-, dichloro-, monochloroacetyl, and acetyl derivatives of four different chiral amines. The study of the enantiomer separation of 11 different crown ethers of 2,2′-binaphthyldiyl showed that the rigidity of the chiral center can be an additional parameter in chiral recognition for the DNBPG phase but not for a β-cyclodextrin bonded chiral phase. It is apparent from this study that steric effects, inductive effects, and molecular rigidity play important roles in chiral recognition with DNBPG chiral stationary phases.     

Structures of the tetrahydrogenated menaquinones fromActinomadura angiospora,Faenia rectivirgula,andSaccharothrix australiensis

The structures of the tetrahydrogenated menaquinones fromActinomadura angiospora, Faenia rectivirgula, andSaccharothrix australiensis were determined by mass spectrometry and proton nuclear magnetic resonance spectrometry. The positions of saturation of the tetrahydrogenated menaquinones fromFaenia rectivirgula andSaccharothrix australiensis were units II plus III (counting from the ring system), whereas that ofActinomadura angiospora had units III and VIII hydrogenated. The tetrahydrogenated menaquinones fromFaenia rectivirgula andSaccharothrix australiensis are similar to those characterized from other Gram-positive taxa to date, whereas that fromActinomadura angiospora represents a hitherto unknown isomer.     

人胸腺上皮细胞培养及其分泌产物

本文通过降低培养基中血清含量,向RPMI 1640培养基中补加三碘甲腺原氨酸而获得一种人胸腺网状上皮细胞占优势生长的培养物。在此培养基中细胞经传代培养长达90天,仍维持正常形态特征。胸腺组织在培养14天后,新生细胞的突起形成网状结构,细胞化学检查和电镜观察表明具有丰富的分泌颗粒,囊泡及张力原纤维束和桥粒等上皮细胞特征。收集合并细胞培养液,经部分纯化后检查其生物活性,表现出具有促进玫瑰花结形成和降低胸腺细胞TdT活性的作用,说明培养细胞的分泌产物具有胸腺激素活性。根据形态学,细胞化学和生物活性检测结果,我们倾向于认为该培养物主要为网状上皮细胞。     

从“湖北光敏感核不育水稻”的未受精子房和花药培养出单倍体植株

采用10种诱导培养基,培养湖北光敏感核不育水稻农垦58品种的未受精子房和花药。共培养未受精子房2790个,获得胚囊愈伤组织17块,最高诱导频率达3.33%,其中2块分化出绿苗。培养花药16740个,获得花药愈伤组织15块,最高诱导频率为0.92%,其中3块分化出苗,2丛白苗,1株绿苗。胚囊植株和花粉植株经根尖染色体检查为单倍体,2n=x=12。实验证明,液体培养、2,4-D0.2-0.5 mg/1、低温预处理对诱导胚囊愈伤组织及花粉愈伤组织的形成具良好效果。     

中国人线粒体DNA的八种限制酶图及其电镜结构

尽管Anderson等人(1981)已测定了人mtDNA的全部序列,但还不能全面地反映整个人类mtDNA核苷酸序列的情况。因此,在具有代表特征的中国人mtDNA序列被测定之前,为了开展对中国人mtDNA的遗传学研究,笔者构建了中国人mtDNA的8种限制酶图。并通过电镜技术对mtDNA进行了研究,发现了一种周长为2μm的小环状DNA,推测它可能在核DNA和mtDNA之间的信息传递或衰老发生中起到某些作用。