Metagenomics to paleogenomics: large-scale sequencing of mammoth DNA

We sequenced 28 million base pairs of DNA in a metagenomics approach, using a woolly mammoth (Mammuthus primigenius) sample from Siberia. As a result of exceptional sample preservation and the use of a recently developed emulsion polymerase chain reaction and pyrosequencing technique, 13 million base pairs (45.4%) of the sequencing reads were identified as mammoth DNA. Sequence identity between our data and African elephant (Loxodonta africana) was 98.55%, consistent with a paleontologically based divergence date of 5 to 6 million years. The sample includes a surprisingly small diversity of environmental DNAs. The high percentage of endogenous DNA recoverable from this single mammoth would allow for completion of its genome, unleashing the field of paleogenomics.     

Mammoth hair: stability of alpha-keratin structure and constituent proteins

Hair from a woolly mammoth (Mammuthus primigenius, age about 32,000 years) retains the ordered structure characteristic of alpha-keratins, but the proteins of this hair differ in composition from and are smaller than similar proteins isolated from other keratins, for example, elephant hair. It is suggested that these changes have been caused by limited proteolysis.     

Antibodies to photoproducts of deoxyribonucleic acids irradiated with ultraviolet light

A rabbit immunized with complexes of methylated bovine serum albumin and ultraviolet-irradiated DNA from calf thymus produced antibodies directed toward the photoproducts in the DNA. Serologic activity appeared after irradiation of DNA at 270 mmicro and decreased upon irradiation at 235 mmicro. The antigenic determinants of the ultraviolet-treated DNA appear to be photoproducts associated primarily with thymine, as measured by direct dependence of serologic activity on the adenine-thymine content of the DNA, and by inhibition of the Serlolgic reaction by the irradiated di-,tri-,and tetra-(thymidine-5'-phosphate nucleotides.     

Identification of an alpha subunit of dihydropyridine-sensitive brain calcium channels

Voltage-sensitive calcium channels in different tissues have diverse functional properties. Polyclonal antibodies (PAC-2) against the alpha subunits of purified rabbit skeletal muscle calcium channels immunoprecipitated calcium channels labeled with the dihydropyridine PN200-110 from both skeletal muscle and brain. The immunoreactivity of PAC-2 with the skeletal muscle channel was greater than that with the brain calcium channel and was absorbed only partially by prior treatment with the brain channel. PAC-2 specifically recognized a large peptide in synaptic plasma membranes of rabbit brain with an apparent molecular size of 169,000 daltons. This protein resembles an alpha subunit of the skeletal muscle calcium channel in apparent molecular weight, antigenic properties, and electrophoretic behavior after reduction of disulfide bonds. Thus, the dihydropyridine-sensitive calcium channel of rabbit brain has an alpha subunit that is homologous, but not identical, to those of the skeletal muscle calcium channel. The different functional properties of these two calcium channels may result from minor variations in structurally similar components.     

Antigenic diversity in the human malaria parasite Plasmodium falciparum

Monoclonal antibodies against blood forms of Plasmodium falciparum were used to demonstrate considerable antigenic diversity in this species. Different isolates were distinguished by their ability to react with certain antibodies, and most of the antibodies reacted specifically with merozoites, schizonts, or both. The distribution of different antigenic types appeared not to be related to geographic origin. Serological typing with monoclonal antibodies extends the range of methods for identification of different strains of this malaria parasite.     

Stimulation in vitro with protein carrier of antibodies against a hapten

Rabbits were immunized both with lysozyme and with dinitrophenylated bovine serum albumin. No antibodies against the dinitrophenyl hapten were produced when cell suspensions of their spleens were exposed in vitro to dinitro-phenyl-ovalbumin, whereas a positive response was obtained with dinitrophenyl-lysozyme. Moreover, when spleen cells from rabbits previously immunized with dinitrophenyl-bovine serum albumin were exposed in vitro to bovine serum albumin alone, they produced antibodies against the dinitrophenyl hapten.     

Individual Antigenic Specificity of Isolated Antibodies

Antisera produced against certain isolated human antibodies showed clear antigenic differences between these antibodies. Individual antigenic specificity was demonstrated for two anti-A antibodies, one anti-dextran, and one anti-levan antibody. Failure to produce specific antisera in other instances appeared to be correlated with a greater heterogeneity of the antibody population used as antigen.     

Intrinsic and extrinsic factors in protein antigenic structure

Recent advances in the preparation of synthetic peptide vaccines and the use of synthetic peptides as probes of antigenic structure and function have led to renewed interest in the prediction of antigenic sites recognized by antibodies and T cells. This review focuses on antibodies. Features intrinsic to the antigen, such as hydrophilicity and mobility, may be useful in the selection of amino acid sequences of the native protein that will elicit antibodies cross-reacting with peptides, or sequences which, as peptides, will be more likely to elicit antibodies cross-reactive with the native protein. Structural mobility may also contribute to protein-protein interactions in general. However, the entire accessible surface of a protein is likely to be detectable by a large enough panel of antibodies. Which of these antibodies are made in any individual depends on factors extrinsic to the antigen molecule, host factors such as self-tolerance, immune response genes, idiotype networks, and the immunoglobulin structural gene repertoire.     

德宏象草不同生育时期营养成分研究

 德宏象草(Pennisetum purpurcum Schumach),在云南德宏分布广泛。营养分析结果说明,在生长的同一时期,该种的粗蛋白、粗脂肪含量高于王草,粗纤维含量低于王草,粗灰分与王草相当,说明在相同生长阶段,德宏象草的质量优于王草。     

不同牛粪施用量对矮象草产草量、营养物质含量及干物质降解率的影响

 【目的】阐明不同牛粪施用量对矮象草产量、营养物质含量及干物质降解率的影响,旨在通过合理施用牛粪提高矮象草的营养价值、降低牛粪中氮、铁、锌和铜等物质对环境的污染。【方法】试验设置不施牛粪（0 kg鲜牛粪/10 kg土）、低量牛粪（3 kg鲜牛粪/10 kg土）和高量牛粪（6 kg鲜牛粪/10 kg土）3组处理,采用盆栽试验和肉牛瘤胃降解试验测定了在不同牛粪施用量下矮象草的产草量、营养物质含量及营养物质降解率的变化。【结果】施用低量和高量牛粪均能极显著的（P＜0.01）提高矮象草的茎和叶干物质产量,其中茎干物质产量分别比不施牛粪提高45.5%和54.5%,叶干物质产量分别提高85.7%和121.4%;施用低量和高量牛粪矮象草叶蛋白质含量和中性洗涤纤维含量也显著地（P＜0.05）高于不施牛粪,其中叶蛋白质含量分别提高13.3%和20.9%。矮象草植株中锌和铜累积能力均表现为：根＞叶＞茎。有机物在瘤胃内尼龙袋中的降解率随牛粪施用量的增加和在瘤胃内的停留时间的延长而增加。【结论】增加牛粪施用量可提高矮象草的产草量、营养物质含量及干物质在瘤胃内的降解率,从而提高矮象草的饲用价值,减少日粮中蛋白质饲料和微量元素添加剂的使用量,但牛粪的适宜施用量还有待于进一步深入研究。     

Serotonin: radioimmunoassay

The development of a radioimmunoassay for serotonin is described. Serotonin is made antigenic by coupling it to bovine serum albumin. Thrombocyte-free plasma instead of serum from the immunized animals is used for radioimmunoassay. Less than 1 ng serotonin can be measured by this procedure.     

Ancient noncoding elements conserved in the human genome

Cartilaginous fishes represent the living group of jawed vertebrates that diverged from the common ancestor of human and teleost fish lineages about 530 million years ago. We generated approximately 1.4x genome sequence coverage for a cartilaginous fish, the elephant shark (Callorhinchus milii), and compared this genome with the human genome to identify conserved noncoding elements (CNEs). The elephant shark sequence revealed twice as many CNEs as were identified by whole-genome comparisons between teleost fishes and human. The ancient vertebrate-specific CNEs in the elephant shark and human genomes are likely to play key regulatory roles in vertebrate gene expression.     

南极凯尔盖朗海台南象海豹繁殖后运动行为特征分析

南象海豹(Mirounga leonina)为南极海洋生态系统中的重要捕食者,研究该生物行为特征在生态学与生物海洋学方面有着重要意义,但针对凯尔盖朗地区南象海豹的区域性行为特点研究则少有论及。基于凯尔盖朗海台北部安置在南象海豹身上的盐度-温度-深度剖面仪(Conductivity-temperature-depth,CTD)所收集的数据,运用切换状态空间模型(Switching state-space model,SSSM)预测了南象海豹的真实水平运动状况,并分析了其觅食时对水团利用情况以及潜水特征。结果表明:水平方向上,食物资源的不均匀分布驱动南象海豹向生产力更高区域迁徙,它们主要聚集在3个生产力较高的区域进行觅食,其中凯尔盖朗海台东部的觅食区为大部分南象海豹的迁徙目标;垂直方向上,南象海豹的潜水行为不受光照水平影响且个体间运动表现差异显著,但具有频次高、深度大的特征,同时将捕食区间限制在冬季水(Winter water, WW)附近,这可能是在捕食对象分布与种间竞争之间进行权衡的结果。研究结果丰富了对凯尔盖朗群岛南象海豹繁殖后运动行为的认知,以期为今后更好地了解海洋捕食者的运动行为和生态特性,从而进一步阐释南极生态系统功能。     

甘蔗的饲用栽培价值评价

以象草为对照,对9份甘蔗材料进行了研究.结果表明:营养期多次刈割利用模式下,甘蔗的再生性、干物质产量显著低于象草.部分甘蔗资源的抗寒性强于象草.甘蔗幼嫩期营养价值差,成熟期优于象草,比象草更适于一次性刈割利用.甘蔗不同材料在早期生长、分蘖、再生性、耐寒性及干物质产量等方面存在一定差异.     

象草根际土壤微生物生理群数量特征研究

[目的]查明象草根际微生物生理群的分布特点。[方法]利用选择性培养基,采用涂抹平板法和稀释法对象草根际土壤微生物生理群（氨化细菌、硝化细菌、反硝化细菌、芽孢杆菌等）数量时空分布特征进行研究。[结果]象草根际各微生物生理群的数量具有明显的根际效应,均为根际〉非根际,但根面与根区相比,根面好气性微生物较多,而根区嫌气性微生物较多;同一根际部位,不同微生物生理类群数量也有差异;象草根面纤维素分解和氮的固定主要是好气性菌起作用,根区纤维素分解主要是嫌气性纤解菌起作用,固氮为好气性自生固氮菌和嫌气性自生固氮菌共同起作用。[结论]象草的根际区域不同,微生物生理群种类不同,其数量时空分布动态也不同。     

Nuclear gene indicates coat-color polymorphism in mammoths

By amplifying the melanocortin type 1 receptor from the woolly mammoth, we can report the complete nucleotide sequence of a nuclear-encoded gene from an extinct species. We found two alleles and show that one allele produces a functional protein whereas the other one encodes a protein with strongly reduced activity. This finding suggests that mammoths may have been polymorphic in coat color, with both dark- and light-haired individuals co-occurring.     

Antigens in Insulin Determinants of Specificity of Porcine Insulin in Man

Porcine insulin, which is distinguished from human insulin only in the amino acid at the C terminal of the B chain, is antigenic in man. Even if the last amino acid or the last eight amino acids are removed from the C terminus of the B chain of insulin, the altered insulin still reacts with human antibodies to porcine insulin; thus, the antigenic determinant of porcine insulin is located in a part of the molecule where the amino acid sequence is the same as it is in the corresponding part of the human insulin molecule.     

Monoclonal antibody to acetylcholine receptor: cell line established from thymus of patient with Myasthenia gravis

A human B cell line producing a monoclonal antibody to an antigenic determinant of acetylcholine receptors was established by cloning B cells that had been transformed in vitro by Epstein-Barr virus. The B cells were obtained from the thymus of a patient with myasthenia gravis. The antibody produced by the cell line precipitated acetylcholine receptors from denervated and innervated rat muscle and from human muscle, but did not show detectable response to the acetylcholine receptors from the electric organs of Narke japonica. The monoclonal antibody showed identical binding patterns in innervated and denervated rat muscles. Passive transfer of the monoclonal antibody into rats induced moderate muscle weakness and electromyographic changes characteristic of myasthenia gravis.     

Poliovirus host range is determined by a short amino acid sequence in neutralization antigenic site I

The mouse-adapted strain of poliovirus type 2 (Lansing) induces fatal poliomyelitis in mice after intracerebral inoculation, whereas mice inoculated with poliovirus type 1 (Mahoney) show no signs of disease. Previous work indicated that the adaptation to mouse virulence is associated with the viral capsid proteins and that mutations in neutralization antigenic site I of poliovirus reduce neurovirulence of the Lansing strain in mice. The role of antigenic site I in mouse neurovirulence was further explored by constructing an antigenic hybrid virus. Six amino acids in antigenic site I of the Mahoney strain were replaced with a sequence specific for the Lansing strain by using a mutagenesis cartridge. The hybrid virus was neutralized by polyclonal antisera elicited by the type 1 and type 2 strains of poliovirus and by neutralizing monoclonal antibodies directed against antigenic site I of type 2 virus. The hybrid virus induced paralytic disease in mice, an observation demonstrating that a short sequence of amino acids in antigenic site I is an important determinant of poliovirus host range. Antigenic site I may be involved in attachment of poliovirus to cells of the mouse central nervous system.