Patient and impatient punishers of free-riders

Costly punishment of cheaters who contribute little or nothing to a cooperating group has been extensively studied, as an effective means to enforce cooperation. The prevailing view is that individuals use punishment to retaliate against transgressions of moral standards such as fairness or equity. However, there is much debate regarding the psychological underpinnings of costly punishment. Some authors suggest that costly punishment must be a product of humans'' capacity for reasoning, self-control and long-term planning, whereas others argue that it is the result of an impulsive, present-oriented emotional drive. Here, we explore the inter-temporal preferences of punishers in a multilateral cooperation game and show that both interpretations might be right, as we can identify two different types of punishment: punishment of free-riders by cooperators, which is predicted by patience (future orientation); and free-riders'' punishment of other free-riders, which is predicted by impatience (present orientation). Therefore, the picture is more complex as punishment by free-riders probably comes not from a reaction against a moral transgression, but instead from a competitive, spiteful drive. Thus, punishment grounded on morals may be related to lasting or delayed psychological incentives, whereas punishment triggered by competitive desires may be linked to short-run aspirations. These results indicate that the individual''s time horizon is relevant for the type of social behaviour she opts for. Integrating such differences in inter-temporal preferences and the social behaviour of agents might help to achieve a better understanding of how human cooperation and punishment behaviour has evolved.     

Fructose 1,6-bisphosphatase: Dissociation of AMP cooperativity and AMP inhibition by carbamylation

Selective treatment of pig kidney fructose 1,6-bisphosphatase with potassium cyanate leads to the formation of an active carbamylated enzyme that has lost the cooperative interactions among AMP sites, but retains sensitivity to inhibition of catalytic activity by the regulator AMP. Incorporation data on [¹⁴C]KNCO indicate that the loss of enzyme cooperativity at the AMP sites is related to selective carbamylation of four lysine residues per mole of tetrameric enzyme. Exhaustive carbamylation suggests that a second lysine residue per subunit is essential for AMP inhibition.     

Enthoprotin: a novel clathrin-associated protein identified through subcellular proteomics

Despite numerous advances in the identification of the molecular machinery for clathrin-mediated budding at the plasma membrane, the mechanistic details of this process remain incomplete. Moreover, relatively little is known regarding the regulation of clathrin-mediated budding at other membrane systems. To address these issues, we have utilized the powerful new approach of subcellular proteomics to identify novel proteins present on highly enriched clathrin-coated vesicles (CCVs). Among the ten novel proteins identified is the rat homologue of a predicted gene product from human, mouse, and Drosophila genomics projects, which we named enthoprotin. Enthoprotin is highly enriched on CCVs isolated from rat brain and liver extracts. In cells, enthoprotin demonstrates a punctate staining pattern that is concentrated in a perinuclear compartment where it colocalizes with clathrin and the clathrin adaptor protein (AP)1. Enthoprotin interacts with the clathrin adaptors AP1 and with Golgi-localized, gamma-ear-containing, Arf-binding protein 2. Through its COOH-terminal domain, enthoprotin binds to the terminal domain of the clathrin heavy chain and stimulates clathrin assembly. These data suggest a role for enthoprotin in clathrin-mediated budding on internal membranes. Our study reveals the utility of proteomics in the identification of novel vesicle trafficking proteins.     

Paratuberculosis in free-ranging fallow deer in Spain

Paratuberculosis was diagnosed in a population of approximately 1,000 free-ranging fallow deer (Dama dama) sampled from 1997-98 in the Regional Hunting Reserve of El Sueve (Asturias, Spain). Five of eight animals observed with diarrhea were diagnosed as having paratuberculosis on the basis of gross lesions at postmortem examination and histopathology. In two deer, Mycobacterium avium subsp. paratuberculosis was cultured and identified by polymerase chain reaction. Indirect enzyme-linked immunosorbent assay and immunodiffusion tests were used to evaluate sera from 33 adult deer from this population. All fallow deer tested were seronegative.     

Topological characterization of the essential Escherichia coli cell division protein FtsW

The membrane topology of Escherichia coli FtsW, a 46-kDa essential protein, was analyzed using a set of 28 ftsW-alkaline phosphatase (ftsW-phoA) and nine ftsW-beta-lactamase (ftsW-bla) gene fusions obtained by in vivo and in vitro methods. The alkaline phosphatase activities or resistance pattern of cells expressing the FtsW-PhoA or FtsW-Bla fusions confirmed only eight out of 10 transmembrane segments predicted by computational methods. After comparison with the recent topology of Streptococcus pneumoniae FtsW, we could identify all the fusions in absolute agreement with the predicted model: N-terminal and C-terminal ends in the cytoplasm, 10 transmembrane segments and one large loop of 67 amino acids (E240-E306) located in the periplasm.     

不同生长阶段青枯菌的高效离子交换色谱表征

采用高效离子交换色谱和激光光散射仪对不同生长阶段的青枯菌进行色谱表征。青枯菌经过色谱分离得到3个特征峰。通过对延滞期、对数生长期和稳定期青枯菌细胞浓度、发酵液pH值、细胞表面黏附的胞外酸性多糖(EPSⅠ)含量与青枯菌色谱行为的关系研究,发现在8h时,青枯菌运动能力强,细胞表面EPSⅠ少,吸附力弱,绝大多数菌体直接随流动相流出形成峰1;随着培养时间延长,细胞表面黏附的EPSⅠ量增多,与树脂吸附力增强,保留时间延长。因此分析3个色谱峰的形成与青枯菌的运动性、以及细胞表面EPSⅠ与阴离子交换树脂的吸附作用有关。并通过比较3个色谱峰青枯菌细胞表面EPSⅠ含量的差异和观察青枯菌经过4%甲醛固定处理后色谱行为的变化加以进一步验证。高效离子交换色谱为细菌完整细胞的研究提供了一种新的分析方法。     

Hybridization-based antibody cDNA recovery for the production of recombinant antibodies identified by repertoire sequencing

High-throughput sequencing of the antibody repertoire is enabling a thorough analysis of B cell diversity and clonal selection, which may improve the novel antibody discovery process. Theoretically, an adequate bioinformatic analysis could allow identification of candidate antigen-specific antibodies, requiring their recombinant production for experimental validation of their specificity. Gene synthesis is commonly used for the generation of recombinant antibodies identified in silico. Novel strategies that bypass gene synthesis could offer more accessible antibody identification and validation alternatives. We developed a hybridization-based recovery strategy that targets the complementarity-determining region 3 (CDRH3) for the enrichment of cDNA of candidate antigen-specific antibody sequences. Ten clonal groups of interest were identified through bioinformatic analysis of the heavy chain antibody repertoire of mice immunized with hen egg white lysozyme (HEL). cDNA from eight of the targeted clonal groups was recovered efficiently, leading to the generation of recombinant antibodies. One representative heavy chain sequence from each clonal group recovered was paired with previously reported anti-HEL light chains to generate full antibodies, later tested for HEL-binding capacity. The recovery process proposed represents a simple and scalable molecular strategy that could enhance antibody identification and specificity assessment, enabling a more cost-efficient generation of recombinant antibodies.     

Ripening in papaya fruit is altered by ACC oxidase cosuppression

Papaya (Carica papaya) is a very important crop in many tropical countries but it is highly susceptible to parasitic diseases, physiological disorders, mechanical damage and fruit overripening. Here we report a study on ACC oxidase cosuppression and its effects on papaya fruit ripening. Papaya ACC oxidase was isolated using PCR and embriogenic cells transformed by biolistic using the CaMV 35S promoter to drive the expression of the PCR fragment in sense orientation. Fifty transgenic lines were recovered and 20 of those were grown under field conditions. Southern analysis showed incorporation of the transgene in different copy numbers in the papaya genome. Fruits were evaluated in terms of texture (firmness), colour development, respiration and ethylene production. A sharp reduction in ethylene and CO2 production was detected, whereas softening and colour development of the peel were also altered. Overall, transgenic fruits showed a delay in ripening rate. A reduction in mRNA level for ACC oxidase in transgenic fruit was clearly detectable by northern blot. More studies are necessary before this technology can be used to extend the shelf life of papaya fruit.