Anaerobic Degradation of Propionate by a Mesophilic Acetogenic Bacterium in Coculture and Triculture with Different Methanogens

A mesophilic acetogenic bacterium (MPOB) oxidized propionate to acetate and CO₂ in cocultures with the formate- and hydrogen-utilizing methanogens Methanospirillum hungatei and Methanobacterium formicicum. Propionate oxidation did not occur in cocultures with two Methanobrevibacter strains, which grew only with hydrogen. Tricultures consisting of MPOB, one of the Methanobrevibacter strains, and organisms which are able to convert formate into H₂ plus CO₂ (Desulfovibrio strain G11 or the homoacetogenic bacterium EE121) also degraded propionate. The MPOB, in the absence of methanogens, was able to couple propionate conversion to fumarate reduction. This propionate conversion was inhibited by hydrogen and by formate. Formate and hydrogen blocked the energetically unfavorable succinate oxidation to fumarate involved in propionate catabolism. Low formate and hydrogen concentrations are required for the syntrophic degradation of propionate by MPOB. In triculture with Methanospirillum hungatei and the aceticlastic Methanothrix soehngenii, propionate was degraded faster than in biculture with Methanospirillum hungatei, indicating that low acetate concentrations are favorable for propionate oxidation as well.     

Sensitization of heat-treated Listeria monocytogenes to added lysozyme in milk.

Listeria monocytogenes was highly resistant to hen egg white lysozyme in whole milk but was sensitive in media and in phosphate buffer. Methods to sensitize the pathogen to lysozyme in milk were investigated. Treatment of whole milk by cation exchange to remove minerals, particularly Ca2+ and Mg2+, slightly promoted inactivation of L. monocytogenes by lysozyme at 4 degrees C over a period of 6 days. Heat treatment (62.5 degrees C for 15 s) strongly sensitized L. monocytogenes to lysozyme in demineralized milk and in MES [2-(N-morpholino)ethanesulfonic acid] buffer. Addition of Ca2+ or Mg2+ to the demineralized milk restored resistance to lysozyme. Cells were more rapidly heat inactivated at 55 degrees C in demineralized milk containing lysozyme, and addition of Ca2+ to the demineralized milk restored the resistance to heat. The results indicate that minerals or mineral-associated components protect L. monocytogenes from inactivation by lysozyme and heat in milk, probably by increasing cell surface stability. The heat treatment of foods containing added lysozyme can probably play a significant role in producing microbiologically safe foods.     

Capsid assembly and involved function analysis of twelve core protein mutants of duck hepatitis B virus.

The roles of different regions of the duck hepatitis B virus (DHBV) core protein on viral capsid assembly and related functions were examined. Twelve deletion and insertion mutations which covered 80% of the DHBV C open reading frame were constructed and expressed in Escherichia coli. The N-terminal region (amino acids 3 to 66) of DHBV core protein was important for its tertiary structure and function in E. coli. The expressed core mutants without this region apparently inhibited E. coli growth. The results of transmission electron microscopy of E. coli thin sections, capsid agarose gel, and sucrose gradient sedimentation demonstrated that a few DHBV core mutants with insertion in the N terminus and deletion in the C terminus retained the ability to form core-like particles in E. coli. However, other mutations in most of N-terminal and central regions strongly inhibited the self-assembly ability of DHBV core protein in E. coli. In addition, the mutant with a C-terminal region deletion (amino acids 181 to 228) lost most of the nucleic acid-binding activity of the DHBV core protein.     

Effects of gamma irradiation on the plasma membrane of suspension-cultured apple cells. Rapid irreversible inhibition of H+-ATPase activity

The effects of ionizing radiation, used in post-harvest treatment of fruit and vegetables. were investigated on cultured apple cells ( Pyrus malus L. cv. Royal Red) on a short-term period. Irradiation (2 kGy) induced an increase of passive ion effluxes from cells and a decrease of cell capacity to regulate external pH. These alterations are likely due to effects on plasma membrane structure and function and were further investigated by studying the effects of irradiation on plasma membrane H⁺-ATPase activity. Plasma membrane-enriched vesicles were prepared and the H⁺-ATPase activity was characterized. Irradiation of the vesicles induced a dose dependent inhibition of H⁺-ATPase activity. The loss of enzyme activity was immediate, even at low doses (0.5 kGy), and was not reversed by the addition of 2m M dithiothreitol. This inhibition may be the result of an irreversible oxidation of enzyme sulfhydryl moieties and/or the result of changes induced within the lipid bilayer affecting the membrane-enzyme interactions. Further analysis of the H⁺-ATPase activity was carried out on vesicles obtained from irradiated cells confirming the previous results. In vivo recovery of activity was not observed within 5 h following the treatment, thus explaining the decrease of cell capacity to regulate external pH.
This rapid irreversible inhibition of the plasma membrane H⁺-ATPase must be considered as one of the most important primary biochemical events occurring in irradiated plant material.     

植物细胞离析酶的制备和应用

用 Aspergillus sp．A-19菌经固体发酵研制成一种新的植物细胞离析酶(SeparatasezA—P)。其离析单细胞的酶活力平均为70 767u／g,有效作用的pH在3．0—7．0,温度为20—45℃。发酵培养基配方是麸皮：桔皮粉：(NH₄)₂SO₄(w／w)为100:100:O．63,料水比为1 :2．0,培养适宜条件为25℃、60小时。     

Human xeroderma pigmentosum group G gene encodes a DNA endonuclease.

Because of defective nucleotide excision repair of ultraviolet damaged DNA, xeroderma pigmentosum (XP) patients suffer from a high incidence of skin cancers. Cell fusion studies have identified seven XP complementation groups, A to G. Previous studies have implicated the products of these seven XP genes in the recognition of ultraviolet-induced DNA damage and in incision of the damage-containing DNA strand. Here, we express the XPG-encoded protein in Sf9 insect cells and purify it to homogeneity. We demonstrate that XPG is a single-strand specific DNA endonuclease, thus identifying the catalytic role of the protein in nucleotide excision repair. We suggest that XPG nuclease acts on the single-stranded region created as a result of the combined action of the XPB helicase and XPD helicase at the DNA damage site.     

MHC antigen expression by melanomas recovered from mice treated with allogeneic mouse fibroblasts genetically modified for interleukin-2 secretion and the expression of melanoma-associated antigens

Recent approaches toward the immunotherapy of neoplastic disease involve the introduction of expression-competent genes for interleukin-2 (IL-2) into autologous malignant cells. Treatment of tumor-bearing experimental animals with the IL-2-secreting cells successfully induces partial and at times complete remissions. In most instances, however, although delayed, progressive tumor growth continues. Here, certain of the characteristic of B16 melanomas (H-2^b) persisting in C57BL/6 mice (H-2^b) treated with an IL-2-secreting, melanoma-antigen-positive cellular immunogen (RLBA-IL-2 cells) are described. Unlike the melanoma cells first injected, B16 cells recovered from mice treated with RLBA-IL-2 cells were deficient in the experssion of MHC class I, but not class II determinants. Deficient MHC class I expression correlated with the cells' resistance to cytotoxic T lymphocytes (CTL) from the spleens of mice immunized with RLBA-IL-2 cells. Melanomas persisting in mice treated with non-IL-2-secreting, melanoma-antigen-positive cell constructs (RLBA-ZipNeo cells) were also deficient in the expression of MHC class I determinants, and the melanoma cells were resistant to CTL from mice immunized with RLBA-ZipNeo cells. Thus, the expression of melanoma-associated antigens rather than IL-2-secretion correlated with deficient MHC class I expression by the persistent melanomas. This point was substantiated by the expression of MHC class I antigens by melanomas persisting in mice treated with IL-2-secreting, melanoma-antigen-negative LM cells (LM-IL-2); it was equivalent to that of melanomas in untreated mice. The involvement of MHC class I antigens in the immune resistance of persistent melanoma cells from mice treated with the melanoma-autigen-positive immunogens was indicated by the effect of interferon (IFN) orN-methyl-N-nitro-N-nitrosoguanidine (MNNG) on the susceptibility of the cells to anti-melanoma CTL. Treatment of the resistant melanomas with IFN or MNNG stimulated MHC class I antigen expression and restored the cells' sensitivity to CTL from mice immunized with IL-2-secreting or nonsecreting, melanoma-antigen-positive cellular immunogens. Prior treatment of the treated cells with antibodies to MHC class I determinants inhibited the cells' susceptibility to CTL from mice immunized with RLBA-IL-2 cells.     

红毛五加叶的三萜皂甙

红毛五加（ＡｃａｎｔｈｏｐａｎａｘｇｉｒａｌｄｉｉＨａｒｍｓ）系五加科五加属植物,分布于河北、河南、四川、陕西、甘肃等省,有祛风湿、通关节、强筋骨、治痿痹等功效［１］。其化学成分未见报道。本文报道红毛五加叶的５个三萜皂甙（甙Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ）的分离鉴定。它们均首次从该属植物发现,其结构如下：　　　　　　　　　　　　　Ｒ１　　　　　　　　　　　　　　Ｒ２　　　　　　　　Ⅰ　ｒｈａｍ－（１→２）－ａｒａ　　　　　Ｈ　　　　　　　Ⅱ　Ｈ　　　ｒｈａｍ－（１→４）－ｇｌｃ－（１→６）－ｇｌｃ　　　　　…     

河西走廊不同生态型芦苇核酸代谢季节动态研究

分布在甘肃河西走廊的４ 种生态型芦苇（Ｐｈｒａｇｍ ｉｔｅｓｃｏｍ ｍ ｕｎｉｓＴｒｉｎ．）的核酸代谢季节变化有差异。盐化草甸芦苇ＲＮＡ 含量持续增加,ＤＮＡ 含量相对稳定,其它３ 种生态型芦苇的ＲＮＡ 和ＤＮＡ 含量以５月份为最高。过渡带芦苇的ＲＮＡ 含量、沼泽芦苇及沙丘芦苇的ＤＮＡ含量９ 月份略有增高。盐化草甸芦苇与过渡带芦苇的ＤＮａｓｅ和ＲＮａｓｅ的活性７ 月份最高,沼泽芦苇与沙丘芦苇的ＤＮａｓｅ和ＲＮａｓｅ活性５—９ 月份呈增高趋势。盐化草甸芦苇的ＤＮＡ 和ＲＮＡ 合成活性不断升高,过渡带芦苇和沼泽芦苇的ＤＮＡ 和ＲＮＡ 合成活性及沙丘芦苇的ＲＮＡ 合成活性５—９月份均降低,仅沙丘芦苇的ＤＮＡ合成活性增强。ＲＮＡ 聚丙烯酰胺梯度凝胶电泳分析结果表明,４ 种生态型芦苇均含有２５Ｓ、２３Ｓ、１８Ｓ、１６Ｓ大分子量ｒＲＮＡ 和小分子量５．８Ｓ、５Ｓ、４．５ＳｒＲＮＡ 及４ＳｔＲＮＡ。大分子量ＲＮＡ 的含量高于小分子量ＲＮＡ 含量。不同生态型及同一生态型芦苇的不同发育时期,相同的ＲＮＡ 组分含量各不相同。且发育过程中２３Ｓ、１８Ｓ、１６ＳｒＲＮＡ 在不同月份发生不同程度的降解。由此,我们认为,核酸代谢的差异性是４ 种生态型由生长转入衰