羟自由基对人红细胞磷脂酰乙醇胺脂质体相变性质的影响

研究了过氧化氢与亚铁离子体系产生的羟自由基对人红细胞膜磷脂酸乙醇胺(PE)脂质体相变性质的影响.结果表明,羟自由基导致脂质体不饱和脂肪酸链的含量明显降低和丙二醛(MDA)含量显著升高,同时其膜流动性随之下降.在室温下,羟自由基诱使PF脂质体冰冻断裂面出现脂质颗粒,说明羟自由基通过脂质过氧化作用可促进PE脂质体从脂双层转变为非双层结构.     

应用PCR对尖锐湿疣和外耳道乳头状瘤中HPV DNA的分型检测

从手术切除的24例女性和12例男性尖锐湿疣新鲜标本中,以及42例女性尖锐湿疣、16例男性外耳道乳头状瘤和4例女性假性湿疣的石蜡包埋标本中,提取组织的基因组DNA,用人工合成的人乳头瘤病毒6.11和16型E6区特异性寡聚核苷酸引物,通过PCR进行HPV DNA的分型检测。结果66例女性尖锐湿疣中,感染HPV6型者4例,感染11型者12例,6+11型混合感染者49例;阴性1例,总检出率达98.4％。4例女性假性湿疣中1例为HPV6型感染,阳性率25％。16例男性外耳道乳头状瘤中HPV6+11型感染者5例,6+16型感染者3例,6+11+16型多重感染者8例,阳性率100％。12例男性尖锐湿疣中,HPV11型感染者7例,6+11型4例,阴性1例,总阳性率91.6％。还对细胞学上空泡化和非典型空泡化尖锐湿疣标本的HPV感染做了比较,未发现差异。     

Orotic acid induces apoptotic death in ovarian adult granulosa tumour cells and increases mitochondrial activity in normal ovarian granulosa cells

Orotic acid (OA) is a natural product that acts as a precursor in the pyrimidine nucleotide biosynthesis pathway. Most studies concerning administration of OA focus on its therapeutic effects; however, its effect on tumours is unclear. We aimed to determine whether treatment with OA influences the viability and apoptosis of normal (HGrC1) and tumour-derived (KGN) human ovarian granulosa cells. The effects of OA (10–250 μM) on viability and apoptosis of both cell lines were determined by using alamarBlue and assessing caspase-3/7 activity, respectively. Annexin V binding and loss of membrane integrity were evaluated in KGN cells. The cell cycle and proliferation of HGrC1 cells were assessed by performing flow cytometric and DNA content analyses, respectively. The influence of OA (10 and 100 μM) on cell cycle- and apoptosis-related gene expression was assessed by RT-qPCR in both cell lines. Mitochondrial activity was analysed by JC-1 staining in HGrC1 cells. In KGN cells, OA reduced viability and increased caspase-3/7 activity, but did not affect mRNA expression of Caspase 3, BAX, and BCL2. OA enhanced proliferation and mitochondrial activity in HGrC1 cells without activating apoptosis. This study demonstrates that the anti-cancer properties of OA in ovarian granulosa tumour cells are not related to changes in apoptosis-associated gene expression, but to increased caspase-3/7 activity. Thus, OA is a promising therapeutic agent for ovarian granulosa tumours. Further, our results suggest that differences in basal expression of cell cycle- and apoptosis-related genes between the two cell lines are responsible for their different responses to OA.     

Variability of plant species diversity in Swedish natural forest and its relation to atmospheric deposition

Within a subprogram of Integrated Monitoring (IM), understorey vegetation in Swedish natural forests was observed at fifteen reference sites over the country for twelve seasons, 1982–1993. The main task of the subprogram was to assess the impact of atmospheric deposition, mainly sulphur and nitrogen, on natural vegetation through time. The present study is focused on the variability of plant species diversity at community level and the possible impact of sulphur and nitrogen deposition. Species richness, evenness and diversity varied greatly among the sites, and between years within each site. Regarding only coniferous forests the species richness was higher in the north than in the south. But the effects of site condition and atmospheric deposition were not clarified. Changes in species diversity through time differed from site to site. No overall temporal trend was found. The atmospheric deposition of sulphur and nitrogen demonstrated a clear geographical pattern being low in the north-west and high in the south-west. Sulphur deposition declined significantly in Southern Sweden during the period. We concluded that the species diversity of understorey vegetation at the Swedish IM sites was not significantly influenced by atmospheric deposition. The changes observed are explained as natural processes.     

生长特性差别的胃癌MGC803细胞亚系间细胞骨架、细胞通讯与癌基因表达的比较研究

肿瘤细胞表型差异是临床药物敏感性、抗原性、转移性、潜伏性和进展速度等差异的主要原因。为了弄清恶性表型差别的表现、相关性及调节机理、我们从胃癌MGC 803系分离了单细胞亚系,各亚系按克隆过程的时间差别归纳分组,选快组M 17、中组M 6和慢组M 3进行比较。软琼脂生长实验表明三者的恶性程度有显著差别;M17恶性度最高,M3最低,M6居中。与其它表型联系比较,证明细胞生长速度、细胞微丝、微管骨架破坏及细胞通讯功能抑制都与锚着不依赖性生长的恶性特征有平行相关性。同时,M 17和M 3原癌基因表达有明显差别。     

Assignment of aliphatic side-chain 1HN/15N resonances in perdeuterated proteins

Summary The perdeuteration of aliphatic sites in large proteins has been shown to greatly facilitate the process of sequential backbone and side-chain ¹³C assignments and has also been utilized in obtaining long-range NOE distance restraints for structure calculations. To obtain the maximum information from a 4D ¹⁵N/¹⁵N-separated NOESY, as many main-chain and side-chain ¹H_N/¹⁵N resonances as possible must be assigned. Traditionally, only backbone amide ¹H_N/¹⁵N resonances are assigned by correlation experiments, whereas slowly exchanging side-chain amide, amino, and guanidino protons are assigned by NOEs to side-chain aliphatic protons. In a perdeuterated protein, however, there is a minimal number of such protons. We have therefore developed several gradient-enhanced and sensitivity-enhanced pulse sequences, containing water-flipback pulses, to provide through-bond correlations of the aliphatic side-chain ¹H_N/¹⁵N resonances to side-chain ¹³C resonances with high sensitivity: NH₂-filtered 2D ¹H-¹⁵N HSQC (H₂N-HSQC), 3D H₂N(CO)C_/ and 3D H₂N(COC_/)C_/ for glutamine and asparagine side-chain amide groups; 2D refocused H(N_/)C_/ and H(N_/C_/)C_/ for arginine side-chain amino groups and non-refocused versions for lysine side-chain amino groups; and 2D refocused H(N)C and nonrefocused H(N_.)C for arginine side-chain guanidino groups. These pulse sequences have been applied to perdeuterated ¹³C-/¹⁵N-labeled human carbonic anhydrase II (²H-HCA II). Because more than 95% of all side-chain ¹³C resonances in ²H-HCA II have already been assigned with the C(CC)(CO)NH experiment, the assignment of the side-chain ¹H_N/¹⁵N resonances has been straightforward using the pulse sequences mentioned above. The importance of assigning these side-chain H_N protons has been demonstrated by recent studies in which the calculation of protein global folds was simulated using only ¹H_N-¹H_N NOE restraints. In these studies, the inclusion of NOE restraints to side-chain H_N protons significantly improved the quality of the global fold that could be determined for a perdeuterated protein [R.A. Venters et al. (1995) J. Am. Chem. Soc., 117, 9592–9593].To whom correspondence should be addressed.     

Expression of a human cytomegalovirus gp58 antigenic domain fused to the hepatitis B virus nucleocapsid protein

Abstract Hepatitis B virus core antigen (HBcAg) has been used as a carrier for expression and presentation of a variety of heterologous viral epitopes in particulate form. The aim of this study was to produce hybrid antigens comprising HBcAg and an immunogenic epitope of human cytomegalovirus (HCMV). A direct comparison was made of amino and carboxyl terminal fusions in order to investigate the influence of position of the foreign epitope on hybrid core particle formation, antigenicity and immunogenicity. HCMV DNA encoding a neutralising epitope of the surface glycoprotein gp58 was either inserted at the amino terminus or fused to the truncated carboxyl terminus of HBcAg and expressed in Escherichia coli . The carboxyl terminal fusion (HBc_3–144-HCMV) was expressed at high levels and assembled into core like particles resembling native HBcAg. Protein with a similar fusion at the amino terminus (HCMV-HBc_1–183) could not be purified or characterised immunologically, although it formed core like particles. HBc_3–144-HCMV displayed HBc antigenicity but HCMV antigenicity could not be detected by radioimmunoassay or western blotting using anti-HCMV monoclonal antibody 7–17 or an anti-HCMV human polyclonal antiserum. Following immunisation of rabbits with HBc_3–144-HCMV, a high titre of anti-HBc specific antibody was produced along with lower titres of HCMV/gp58 specific antibody.     

Anti-endotoxin antibodies directed against Escherichia coli R-1 oligosaccharide core-tetanus toxoid conjugate bind to smooth, live bacteria and smooth lipopolysaccharides and attenuate their tumor necrosis factor stimulating activity

Abstract A hybridoma cell line producing a human anti-lipid A monoclonal antibody (mAb), FKF-1F3 (IgM (κ)) was obtained by cell fusion of Epstein-Barr virus-transformed cells and mouse myeloma. The mAb bound to not only Gram-negative bacterial lipid A, but also to polysaccharide portions of Pseudomonas aeruginosa lipopolysaccharides (LPS). The mAb seemed to recognize two distinct regions of P. aeruginosa LPS other than lipid A, namely the outer core regions of some serotype strains and the O -polysaccharide region of serotype A strains. The mAb cross-reacted with N -acetyl-β-glucosamine-conjugated bovine serum albumin, N -acetyl-β-galactosamine-conjugated bovine serum albumin, myosin and actin, but not with other autoantigens such as ss- and ds-DNA, cardiolipin and glycosaminoglycans. The mAb conferred protective activity against a mouse pseudomonal infection model. The evidence suggested that the mAb was a naturally occurring polyspecific antibody that participated in defense against pseudomonal infections.     

Gathered fruits and cultivated plants at Bercy (Paris), a Neolithic village in a fluvial context

Recent rescue excavations at Bercy (Paris), a site on the alluvial plain of the Seine valley, yielded plant remains which are associated with the recent occupation phase dating from the middle Neolithic II (Chasséen), when a village was established on the former channel of the river Seine. Various contexts (the channel, the flat lower part of the bank, and several archaeological features) have been studied and 84 taxa have been identified. Cultivated plants are represented by Triticum aestivum/durum, T. dicoccum, T. monococcum and Hordeum vulgare. Among the wild plants with potentially edible fruits or seeds, only very few satisfy the various criteria for association with human activities: Corylus avellana, Vitis sylvestris, Cornus sanguinea, Quercus sp. and Prunus spinosa of which carbonized fruits were also present, were found in archaeological features and were very abundant. Though not found in archaeological features, we consider that Crataegus monogyna (carbonized and well represented) and Rubus spp. (especially abundant) were not deposited there naturally and had also been intentionally collected. Finally, it is suggested that the allochthonous (varied) origin of these taxa is the reason why there are no concentrations of their fruits in the channel. The exploitation of wild seeds and fruits appears to have been very selective. All other wild taxa can be attributed to natural deposition.