Temperature and dose effects on the pathogenicity and reproduction of two Korean isolates of Heterorhabditis bacteriophora (Nematoda: Heterorhabditidae)

Out of some isolated Heterorhabditis bacteriophora from Korea, ecological study on two isolates which had different geographical features was investigated. That is, effects of temperature and dose on the pathogenicity and reproduction of two Korean isolates of H. bacteriophora were investigated using Galleria mellonella larvae in the laboratory. The median lethal dose (LD₅₀) decreased with increasing temperature, but increased at 35 °C. The optimal temperatures for infection were 30 °C for H. bacteriophora Jeju strain and 24 °C for H. bacteriophora Hamyang strain. The median lethal time, LT₅₀ of H. bacteriophora Hamyang strain was recorded at 13 °C to 35 °C and that of H. bacteriophora Jeju strain was recorded at 18 °C to 30 °C. The number of established nematodes in G. mellonella larvae was significantly different depending on temperature and dose. When G. mellonella larvae were exposed to 300 infective juveniles (IJs), mortality of G. mellonella gradually increased with exposure time with H. bacteriophora Jeju strain but not with H. bacteriophora Hamyang strain. 87.5% mortality of G. mellonella was recorded by H. bacteriophora Hamyang strain after 1440 min whereas 100% mortality was recorded by H. bacteriophora Jeju strain after 4320 min. The time from infection to the first emergence of nematodes decreased with increasing temperature. Duration of emergence of the two strains in the White traps also decreased with increasing temperature. The highest progeny numbers of H. bacteriophora Jeju strain were 264,602 while those of H. bacteriophora Hamyang strain were 275,744 at the rate of 160 IJs at 24 °C.     

Triad 1 induces apoptosis by p53 activation

Triad 1 (2 RING [really interesting new gene] fingers and DRIL [double RING finger linked] 1) is an E3 ligase that induces apoptosis and clonogenic inhibition in myeloid cells through Gfi-1 stabilization. Here we demonstrate that Triad 1 induces apoptosis in several cancer cell lines including MCF7, A549, U2OS, and HCT 116 p53^+/+ cells via its RING ligase activity. Interestingly, in these cancer cells, Triad 1-induced apoptosis is not mediated by Gfi-1 stabilization but is instead p53-dependent. Moreover, Triad 1 promotes transactivation of p53. These results suggest that Triad 1 can induce apoptosis through its ligase activity via p53 activation.     

Increased ethanol resistance in <Emphasis Type="Italic">Ethanolic Escherichia coli</Emphasis> by Insertion of heat-shock genes BEM1 and SOD2 from <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Ethanol is generally toxic to microorganisms, and intracellular and extracellular accumulation of ethanol inhibits cell growth and metabolism. In this study, pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB) were cloned into pET-32a vector and then introduced into E. coli BL21 to produce ethanol. Heat shock genes (BEM1 and SOD2) from Saccharomyces cerevisiae were inserted into recombinant ethanolic E. coli using pET28_a vector to improve ethanol shock resistance. Three different strains were constructed: Ethanolic E. coli (adhB and pdc genes inserted using pET32_a vector), BEM1 gene-inserted E. coli (BEM1 inserted using pET_28a), and SOD2-inserted E. coli (SOD2 inserted using pET28_a). Construction of these three different strains allowed comparison of the functions of these heat shock genes as well as their roles in ethanol tolerance. The toxicity of ethanol in recombinant ethanolic E. coli was tested by measuring cell growth in response to various ethanol concentrations. The results show that SOD2-inserted E. coli showed higher ethanol resistance than ethanolic E. coli.     

Alkali‐based AFEX pretreatment for the conversion of sugarcane bagasse and cane leaf residues to ethanol

Sugarcane is one of the major agricultural crops cultivated in tropical climate regions of the world. Each tonne of raw cane production is associated with the generation of 130 kg dry weight of bagasse after juice extraction and 250 kg dry weight of cane leaf residue postharvest. The annual world production of sugarcane is ～1.6 billion tones, generating 279 MMT tones of biomass residues (bagasse and cane leaf matter) that would be available for cellulosic ethanol production. Here, we investigated the production of cellulosic ethanol from sugar cane bagasse and sugar cane leaf residue using an alkaline pretreatment: ammonia fiber expansion (AFEX). The AFEX pretreatment improved the accessibility of cellulose and hemicelluloses to enzymes during hydrolysis by breaking down the ester linkages and other lignin carbohydrate complex (LCC) bonds and the sugar produced by this process is found to be highly fermentable. The maximum glucan conversion of AFEX pretreated bagasse and cane leaf residue by cellulases was ～85%. Supplementation with hemicellulases during enzymatic hydrolysis improved the xylan conversion up to 95–98%. Xylanase supplementation also contributed to a marginal improvement in the glucan conversion. AFEX‐treated cane leaf residue was found to have a greater enzymatic digestibility compared to AFEX‐treated bagasse. Co‐fermentation of glucose and xylose, produced from high solid loading (6% glucan) hydrolysis of AFEX‐treated bagasse and cane leaf residue, using the recombinant Saccharomyces cerevisiae (424A LNH‐ST) produced 34–36 g/L of ethanol with 92% theoretical yield. These results demonstrate that AFEX pretreatment is a viable process for conversion of bagasse and cane leaf residue into cellulosic ethanol. Biotechnol. Bioeng. 2010;107: 441–450. © 2010 Wiley Periodicals, Inc.     

Identification of the catalytic residues involved in the carboxyl transfer of pyruvate carboxylase

To clarify the mechanism of carboxyl transfer from carboxylbiotin to pyruvate, the following conserved amino acid residues present in the carboxyl transferase domain of Bacillus thermodenitrificans pyruvate carboxylase were converted to homologous amino acids: Asp543, Glu576, Glu592, Asp649, Lys712, Asp713, and Asp762. The carboxylase activity of the resulting mutants, D543E, E576D, E576Q, E592Q, D649N, K712R, K712Q, D713E, D713N, D762E, and D762N, was generally less than that of the wild type from mutation, but it decreased the most to 5% or even less than that of the wild type with D543E, D576Q, D649N, K712R, and K712Q. The decrease in activity observed for Asp543, Asp649, and Lys712 mutants was not for structural reasons because their structures seemed to remain intact as assessed by gel filtration and circular dichroism. On the basis of these data, a mechanism is proposed where Lys712 and Asp543 serve as the key acid and base catalyst, respectively.     

Expression of Expanded Polyglutamine Protein Induces Behavioral Changes in Drosophila (Polyglutamine-Induced Changes in Drosophila)

Spinocerebellar ataxia type-3 or Machado-Joseph disease (SCA3/MJD) is an autosomal dominant neurodegenerative disease caused by triplet nucleotide expansion. The expansion of the polyglutamine tract near the C terminus of the MJD1 gene product, ataxin-3, above a threshold of 40 glutamine repeats causes neuronal loss and degeneration. The expanded ataxin-3 forms aggregates, and nuclear inclusions, within neurons, possibly due to the misfolding of mutant proteins. Here we report upon the behavioral test changes related to truncated and expanded forms of MJD protein (MJDtr) in Drosophila, and show that expanded MJDtr, when expressed in the nervous system, causes characteristic locomotor dysfunction and anosmia. This phenomenon has not been previously reported in humans or in transgenic Drosophila models. In addition, the in vivo expression of the antiapoptotic gene bcl-2 showed no evidence of ameliorating the deleterious effect of MJDtr-Q78s, either in the eye or in the nervous system. The study shows that such Drosophila transgenic models express olfactory dysfunction and ataxic behavior as observed in human patients.     

Daily short-period gravitation can prevent functional and structural changes in arteries of simulated microgravity rats.

This study was designed to clarify whether simulated microgravity-induced differential adaptational changes in cerebral and hindlimb arteries could be prevented by daily short-period restoration of the normal distribution of transmural pressure across arterial vasculature by either dorsoventral or footward gravitational loading. Tail suspension (Sus) for 28 days was used to simulate cardiovascular deconditioning due to microgravity. Daily standing (STD) for 1, 2, or 4 h, or +45 degrees head-up tilt (HUT) for 2 or 4 h was used to provide short-period dorsoventral or footward gravitational loading as countermeasure. Functional studies showed that Sus alone induced an enhancement and depression in vasoconstrictor responsiveness of basilar and femoral arterial rings, respectively, as previously reported. These differential functional alterations can be prevented by either of the two kinds of daily gravitational loading treatments. Surprisingly, daily STD for as short as 1 h was sufficient to prevent the differential functional changes that might occur due to Sus alone. In morphological studies, the effectiveness of daily 4-h HUT or 1-h STD in preventing the differential remodeling changes in the structure of basilar and anterior tibial arteries induced by Sus alone was examined by histomorphometry. The results showed that both the hypertrophic and atrophic changes that might occur, respectively, in cerebral and hindlimb arteries due to Sus alone were prevented not only by daily HUT for 4 h but also by daily STD even for 1 h. These data indicate that daily gravitational loading by STD for as short as 1 h is sufficient to prevent differential adaptational changes in function and structure of vessels in different anatomic regions induced by a medium-term simulated microgravity.     

小麦不同生育时期Cd、Cr、Pb污染监测指标体系

以小麦为供试材料,分别采用发芽试验、溶液培养、土柱栽培等毒理试验方法,研究了小麦萌芽期、幼苗期及成株期受重金属污染毒害的指标体系。结果表明,①小麦根伸长抑制率可作为萌芽期重金属污染评价的一项生物指标;Cd、Cr、Pb对小麦根生长的7d半效应浓度(EC50)值分别为1.39、0.20mmo.lL-1和2.75mmol.L-1,据此得到3种重金属对小麦同一性状的毒性次序为CrCdPb;此外各性状抑制率与胁迫浓度的关系可用双曲线模型y=x/(a+x/100)或指数曲线模型y=a(1-exp(-bx))较好模拟。②Cd、Cr、Pb单一污染胁迫对小麦幼苗性状的毒性次序均以影响叶面积和冠部干重为主,表明叶面积和冠部干重为幼苗期污染监测的敏感指标;重金属对除根干重外所有性状的毒性次序:CrCdPb,该毒性次序是根据引起50%抑制的临界浓度即EC50值确定的;各性状抑制率与胁迫浓度的关系可用直线模型、或双曲线模型、或指数曲线模型较好模拟。③重金属单一污染胁迫造成小麦籽粒产量下降幅度是CdCrPb;二元互作对产量影响的重要性次序为CdCrCdPbCrPb;在Cd、Cr、Pb复合污染胁迫条件下,穗数可以作为成株期重金属污染监测的首选指标。以上指标可为农田小麦重金属污染的监测和综合治理提供一些理论依据。