Crystal structure and molecular dynamics studies of purine nucleoside phosphorylase from Mycobacterium tuberculosis associated with acyclovir

Consumption has been a scourge of mankind since ancient times. This illness has charged a high price to human lives. Many efforts have been made to defeat Mycobacterium tuberculosis (Mt). The M. tuberculosis purine nucleoside phosphorylase (MtPNP) is considered an interesting target to pursuit new potential inhibitors, inasmuch it belongs to the purine salvage pathway and its activity might be involved in the mycobacterial latency process. Here we present the MtPNP crystallographic structure associated with acyclovir and phosphate (MtPNP:ACY:PO₄) at 2.10 Å resolution. Molecular dynamics simulations were carried out in order to dissect MtPNP:ACY:PO₄ structural features, and the influence of the ligand in the binding pocket stability. Our results revealed that the ligand leads to active site lost of stability, in agreement with experimental results, which demonstrate a considerable inhibitory activity against MtPNP (K_i = 150 nM). Furthermore, we observed that some residues which are important in the proper ligand’s anchor into the human homologous enzyme do not present the same importance to MtPNP. Therewithal, these findings contribute to the search of new specific inhibitors for MtPNP, since peculiarities between the mycobacterial and human enzyme binding sites have been identified, making a structural-based drug design feasible.     

外源亚精胺和精胺对NaHCO3胁迫下南蛇藤抗氧化系统的影响

研究了叶面喷施亚精胺和精胺对NaHCO₃胁迫下南蛇藤叶片抗氧化系统的影响.结果表明：外源亚精胺和精胺处理使NaHCO₃胁迫下南蛇藤叶片O₂^-·产生速率、H₂O₂、丙二醛(MDA)含量和电解质外渗率显著降低(P<0.05).亚精胺处理明显提高了盐胁迫下超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)等抗氧化酶的活性,以及还原型谷胱甘肽(GSH)、类胡萝卜素(CAR)和脯氨酸(Pro)等抗氧化剂的含量,但对还原型抗坏血酸(AsA)含量没有作用;精胺处理明显提高NaHCO₃胁迫下POD和APX的活性以及GSH、CAR和Pro的含量,但对SOD和AsA含量影响不显著,甚至引起CAT活性明显降低.亚精胺和精胺处理明显改善了NaHCO₃胁迫下南蛇藤的生长.外源亚精胺和精胺可以改善NaHCO₃胁迫下南蛇藤叶片的膜保护功能,减少叶片中活性氧的积累,从而提高南蛇藤对NaHCO₃胁迫的抗性.     

Antibodies designed as effective cancer vaccines

Antigen/antibody complexes can efficiently target antigen presenting cells to allow stimulation of the cellular immune response. Due to the difficulty of manufacture and their inherent instability complexes have proved inefficient cancer vaccines. However, anti-idiotypic antibodies mimicking antigens have been shown to stimulate both antibody and T cell responses. The latter are due to T cell mimotopes expressed within the complementarity-determining regions (CDRs) of antibodies that are efficiently presented to dendritic cells in vivo. Based on this observation we have designed a DNA vaccine platform called ImmunoBody™, where cytotoxic T lymphocyte (CTL) and helper T cell epitopes replace CDR regions within the framework of a human IgG1 antibody. The ImmunoBody™ expression system has a number of design features which allow for rapid production of a wide range of vaccines. The CDR regions of the heavy and light chain have been engineered to contain unique restriction endonuclease sites, which can be easily opened, and oligonucleotides encoding the T cell epitopes inserted. The variable and constant regions of the ImmunoBody™ are also flanked by restriction sites, which permit easy exchange of other IgG subtypes. Here we show a range of T cell epitopes can be inserted into the ImmunoBody™ vector and upon immunization these T cell epitopes are efficiently processed and presented to stimulate high frequency helper and CTL responses capable of anti-tumor activity.Key words: DNA vaccines, cancer vaccines, melanoma, CTL, helper T cells     

Adaptive response to H(2)O(2) protects against SnCl(2) damage: the OxyR system involvement

The stannous ion, mainly the stannous chloride (SnCl(2)) salt form, is widely used as a reducing agent to label radiotracers with technetium-99m ((99m)Tc). These radiotracers can be employed as radiopharmaceuticals in nuclear medicine procedures. In this case, there is no doubt about absorption of this complex, because it is intravenously administered in humans, although biological effects of these agents have not been fully understood. In this work we used a bacterial system to study the cytotoxic potential of stannous chloride. It is known that SnCl(2) induces lesions that could be mediated by reactive oxygen species (ROS). We, thus, investigated the existence of cross-adaptive response between hydrogen peroxide (H(2)O(2)) and SnCl(2) and the role of the OxyR system known to promote cellular protection against oxidative damages. Here we describe the results obtained with prior treatment of different Escherichia coli strains with sub-lethal doses of H(2)O(2), followed by incubation with SnCl(2). Our data show that H(2)O(2) is capable of inducing cross-adaptive response against the lethality promoted by SnCl(2), suggesting the OxyR system participation through catalase, alkyl hydroperoxide reductase and superoxide dismutase enzymes     

The construction of the first balancer chromosome for the Mediterranean fruit fly, Ceratitis capitata

The construction of the first balancer chromosome, FiM1, for the medfly Ceratitis capitata is described. This chromosome has three overlapping pericentric inversions and is marked with dominant and recessive mutations. The inversion breakpoints of FiM1 suppress recombination throughout the length of the fifth chromosome, allowing lethal mutations to be recovered and maintained. This chromosome will provide a powerful tool for the manipulation of laboratory stocks, in particular, the recovery of new mutant and transgenic strains. We demonstrate the use of FiM1 for the recovery and maintenance of chromosomes carrying lethal mutations.     

Mass rearing of temperature sensitive genetic sexing strains in the Mediterranean fruit fly (Ceratitis capitata)

Genetic sexing strains (GSS) based on the temperature sensitive lethal(tsl) mutation are being used to produce sterile male medflies for large scale sterile insect technique (SIT) programmes for this pest. The use of male-only strains increases the overall efficiency of the technique. Currently more than 1.4 billion sterile male-only pupae are produced per week in different facilities around the world. Due to the mutations used to construct these strains, that is, translocations and selectable markers, they require different and more careful mass rearing procedures than do bisexual strains (BSS). The basic rearing technology has been developed and can be used to produce only males on a predictable basis to a level of 99.9% accuracy. If specific rearing procedures are followed, then tsl-based GSS has a rearing efficiency that is equal to that of a BSS and it is already know that males produced by the tsl-based GSS are of equal quality to males produced by BSS. Based on current rearing technology the cost of production of male pupae is about the same for both types of strain. This is due to the large colony that is required for the tsl-based GSS. This paper discusses the considerations that need to be taken into account during mass rearing of GSS and identifies the most efficient production processes that are currently available.     

Photosynthetic generation of O2 and H2 by photosystem I-deficient chlamydomonas mutants

A comparative study of aerobic generation of O2 and anaerobic photoproduction of H2 in whole cells of a wild-type strain of Chlamydomonas reinhardtii and its photosystem I-deficient mutants B4 and F8 found no contribution of photosystem II to ferredoxin photoreduction, which is not consistent with data of recent studies by Greenbaum et al. (Nature, 1995, 376, 438-441; and Science, 1996, 273, 364-367) who reported that they had discovered such a capacity in these mutant strains. In the wild-type and mutant strains, action spectra showed that O2 was evolved by photosystem II, whereas photoinhibition of chlororespiration and evolution of H2 depended on the activity of photosystem I. Single-turnover flash measurements of H2 evolution showed that the contents of photosystem I in mutant strains amounted to 3-35% of that in the wild-type strain. This fraction of photosystem I in "leaky" mutants displayed abnormal kinetic features and was highly sensitive to photoinhibition.     

Genome size variations within Dasypyrum villosum: correlations with chromosomal traits, environmental factors and plant phenotypic characteristics and behaviour in reproduction

 Feulgen/DNA cytophotometric determinations carried out in the root meristem of seedlings showed that substantial quantitative alterations in the nuclear genome are present between and within 15 natural populations of Dasypyrum villosum in Italy. When the most variant values are considered, there is a 17.6% difference between the mean genome size of the populations, and a 66.2% difference between the genome size of individual plants within a population. A highly significant, positive correlation was found to exist between the genome size of D. villosum plants and the altitude of their stations, and differences in DNA contents between individual plants were greater in populations from mountain sites. Karyological analyses showed all chromosome pairs to differ largely in size between plants with differing DNA contents. A highly significant, positive correlation was found to exist between genome size and both the length of the chromosome complement at metaphase and the length and arm ratio of pair VII. Significant correlations were also found between DNA content and certain phenotypic characteristics of the plants. The mean genome size of the populations was negatively correlated with the mean leaf length and width. In contrast, the genome size of individual plants was positively correlated with the weight of the seed from which they originated and their flowering interval. A large range of genome sizes was found in the half-sib progeny of a plant having a relatively large genome. In contrast, in the half-sib progeny of a plant having a small genome, the genome sizes of the individual plants were less divergent and similar to that of the mother plant. All siblings from crosses between plants with differing genome sizes had similar DNA contents, which were intermediate between those of the parental plants, even if closer to the DNA content of the parent plant having the smaller genome size. Size polymorphism within pairs was never observed in plants obtained from these crosses or in half-sibs whose genome size differed from that of the mother plant. The intraspecific alterations observed in the nuclear genome and their effects on plant development and phenotype are briefly discussed as evolutionary factors which allow D. villosum populations to withstand different environmental conditions as well as the variability of conditions in a given environment. Received: 6 October 1997 / Accepted: 28 October 1997     

Changes in contraceptive use and fertility: El Salvador, 1978-88

In El Salvador from 1978 to 1988, contraceptive use among married women 15-44 years of age increased from 34% to 47%, and the total fertility rate declined from 6.3 to 4.6 children per woman. Most of this change took place from 1978 to 1985. Sterilization is the most prevalent method used, but nearly one-half of the women who are sterilized did not use any contraception before their operation. Few young couples use reversible methods of contraception to space births or delay the start of childbearing. On average, women wait 8 years after marriage and have nearly three children before they use contraception.