Comparative studies on beta-glucan hydrolases. Isolation and characterization of an exo(1 yields 3)-beta-glucanase from the snail, Helix pomatia.

An exo-β-glucan hydrolase, present in the digestive juice of the snail, Helix pomatia, has been purified to homogeneity by chromatography on Bio-Gel P-60, Sephadex G-200, DEAE-cellulose, and DEAE-Sephadex. The enzyme degrades β-(1 → 3)-linked oligosaccharides and polysaccharides, rapidly and to completion, or near completion, yielding glucose as the major product of enzyme action. Mixed linkage (1→3; 1→4)-β-glucans are also extensively degraded and β-(1→6)- and β-(1→4)-linked glucose polymers are slowly degraded by the enzyme. This enzyme differs from other exo-β-glucanases, reported previously, in the broadness of its substrate specificity. The K_m values for action on laminarin and lichenin are respectively 1.22 and 2.22 mg/ml; the maximum velocity of action on laminarin is approximately twice that on lichenin. The enzyme has a molecular weight of 82,000 as determined by polyacrylamide gel electrophoresis. Maximum activity is exhibited at pH 4.3 and at temperatures of 50–55 °C.     

In vivo metabolism of 3H-testosterone in adult male rats: effects of estrogen administration.

The metabolism of testosterone (T) was studied in normal adult male rats using a constant infusion of trace amounts of the 3H-steroid into a tail vein for 3 h in order to attain a state of equilibrium. Samples of plasma, liver, kidney, prostate, seminal vesicles and muscle were analysed for 3H-testosterone, 3H-5alpha-dihydrotestosterone (5alphaDHT) and 3H-5alpha-androstanediol (Adiol). When compared to the 3H-T level in plasma there were high levels of 3H-T in kidney and of 3H-5alphaDHT in prostate and seminal vesicles. Intraperitoneal estradiol valerate administration (100 mug/day) for 4 days decreased and 3H-5alphaDHT levels in the prostate and seminal vesicles. The estrogen administration increased the T metabolic clearance rate from 17.5 1/24 h/100 g body wt to 22.6 1/24 h/100 g body wt.     

Biotransformation of pregnenolone - 7alpha-3H, progesterone - 7alpha-3H and dehydroepiandrosterone - 7alpha-3H by porcine fetal and maternal adrenal homogenate preparations.

The biotransformation of pregnenolone-7alpha-3H and of progesterone-7alpha-3H by porcine fetal and maternal adrenal homogenates at 56 and 112 days of pregnancy and of dehydroepiandrosterone-7alpha-3H by fetal adrenal homogenates has been investigated in vitro. Both pregnenolone-7alpha-3H and progesterone-7alpha-3H were metabolized extensively by maternal adrenal preparations, the principal radioactive metabolites isolated being cortisol, corticosterone, 11-deoxycortisol, deoxycorticosterone, 11beta-hydroxyprogesterone and androstenedione. In addition, 17alpha-hydroxyprogesterone, 20alpha-dihydroprogesterone and cortisone were formed from both substrates and 17alpha-hydroxypregnenolone and progesterone were formed from pregnenolone. Although essentially the same radioactive metabolites were isolated after incubation of fetal adrenal glands with pregnenolone-7alpha-3H or progesterone-7alpha-3H, a greater proportion of the radioactivity was associated with corticosteroids at 112 days of pregnancy than at 56 days. 11beta-Hydroxyandrostenedione and androstenedione were isolated and identified together with an unknown polar metabolite, after incubation of fetal adrenal tissue with dehydroepiandrosterone-7alpha-3H. These results are discussed in relation to feto-placental steroid biosynthesis and metabolism and the role of the fetal adrenal in the initiation of parturition in the pig.     

丛枝菌根真菌对褐土玉米氮素吸收和土壤N2O排放的影响

探究不同氮肥水平下丛枝菌根(AM)真菌对褐土玉米土壤N₂O排放和氮转化功能基因的影响,为阐明AM真菌在褐土N₂O排放中的作用和效应提供理论依据。设置氮肥用量(NⅠ:105 mg/kg;NⅡ:210 mg/kg)、AM真菌(M0:不接种AM真菌;M1:接种根内根孢囊霉(Rhizophagus intraradices);M2:接种摩西斗管囊霉(Funneliformis mosseae);M3:接种Rhizophagus intraradices + Funneliformis mosseae等比例混合)双因素盆栽试验。测定植株地上部全氮含量、土壤铵态氮、硝态氮含量和N₂O排放量,采用实时荧光定量聚合酶链式反应(PCR)法分析土壤硝化功能基因(amoA-AOA和amoA-AOB)和反硝化功能基因(nirS、nirK和nosZ)的丰度。结果表明,两种施氮水平下,接种AM真菌均可显著降低土壤N₂O排放通量和累积排放量,不同AM真菌处理下N₂O累积排放量表现为:M0>M2>M1>M3。相同AM真菌处理的土壤N₂O排放通量和累积排放量在NⅡ施氮水平高于NⅠ施氮水平;相同AM真菌处理的玉米菌根侵染率在NⅡ施氮水平低于NⅠ施氮水平。与M0相比,NⅠ条件下M1、M2和M3处理土壤铵态氮含量分别降低24.5%、20.8%和45.3%,硝态氮含量分别降低19.7%、14.9%和30.2%,植株地上部全氮含量分别增加16.3%、35.2%和59.6%;与M0相比,NⅡ条件下M1、M2和M3处理土壤铵态氮含量分别降低20.9%、24.8%和40.0%,硝态氮含量分别降低36.3%、25.6%和45.2%,植株地上部全氮含量分别增加33.2%、43.9%和95.4%。两种施氮水平下,AM真菌可显著降低土壤硝化功能基因(amoA-AOA和amoA-AOB)丰度,增加反硝化功能基因(nirS、nirK和nosZ)丰度。AM真菌与N₂O排放通量呈极显著负相关。本盆栽试验条件下,接种AM真菌均可增强两种氮肥用量玉米植株氮素吸收能力,调节硝化、反硝化相关功能基因的丰度,减少土壤N₂O气体的排放,且两种AM真菌混合处理的N₂O减排效应强于单一AM真菌接种。     

连翘花的结构与繁育系统研究

通过定株观察、解剖和人工套袋交叉授粉试验对连翘花的生长发育和繁殖系统进行研究.结果表明:(1)连翘具有2种避免自交的方式,雌雄异位和雌雄异熟.其雌雄异位表现为长雄蕊短雌蕊花型和短雄蕊长雌蕊花型;雌雄异熟表现为长雄蕊短雌蕊花型为雄蕊先于雌蕊成熟,短雄蕊长雌蕊花型表现为雌蕊先于雄蕊成熟.(2)连翘花P/O值测定和户外套袋交叉授粉试验显示,P/O值为2 000±300;不同类型花的异花授粉结果率在50.64%~80.32%,其中短型花的花粉授到长型花柱头的结果率最高,达80.32%,而长型花和短型花的同型花授粉结果率分别为2.92%和34.15%,表明连翘的异花授粉结果率高于自花授粉,以异交为主,其繁育系统属兼性异交.研究结果认为,连翘雌雄异位和雌雄异熟是其自然结果率低下的主要原因,为进一步探讨连翘在木犀科中的系统进化提供了生殖生态学的依据.     

Inactivation of Escherichia coli O157:H7 and Salmonella on artificially or naturally contaminated mung beans (Vigna radiata L) using a stabilized oxychloro-based sanitizer

AIMS: To evaluate the efficacy of a stabilized oxychloro-based (SOC) sanitizer to decontaminate mung beans artificially or naturally contaminated with Escherichia coli O157:H7 or Salmonella. METHODS AND RESULTS: Naturally contaminated beans were produced by introducing a five-strain cocktail of E. coli O157:H7 or Salmonella onto the flowers of growing mung bean plants. Escherichia coli O157:H7 was only sporadically recovered from sprout lots (three testing positive from 10 tested) derived from harvested beans. In contrast, Salmonella was recovered from 18 of 20 lots screened. Pathogens present on naturally contaminated seed could be successfully inactivated with SOC applied at 200 ppm for 24 h at 28 degrees C. SOC treatment could also decontaminate artificially inoculated mung bean batches containing different levels of contaminated seed. SOC inactivated E. coli O157:H7, but not Salmonella introduced onto damaged (scarified) beans. CONCLUSIONS: SOC sanitizer could inactivate Salmonella or E. coli O157:H7 naturally or artificially introduced onto mung beans. However, the SOC treatment failed to inactivate Salmonella introduced onto damaged mung beans. SIGNIFICANCE AND IMPACT OF THE STUDY: SOC sanitizer represents an effective method for decontaminating undamaged mung beans.     

Reactive oxygen and nitrogen species during meiotic resumption from diplotene arrest in mammalian oocytes

Mammalian ovary is metabolically active organ and generates by‐products such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) on an extraordinary scale. Both follicular somatic cells as well as oocyte generate ROS and RNS synchronously and their effects are neutralized by intricate array of antioxidants. ROS such as hydrogen peroxide (H₂O₂) and RNS such as nitric oxide (NO) act as signaling molecules and modulate various aspects of oocyte physiology including meiotic cell cycle arrest and resumption. Generation of intraoocyte H₂O₂ can induce meiotic resumption from diplotene arrest probably by the activation of adenosine monophosphate (AMP)‐activated protein kinase A (PRKA)—or Ca²⁺‐mediated pathway. However, reduced intraoocyte NO level may inactivate guanylyl cyclase‐mediated pathway that results in the reduced production of cyclic 3′,5′‐guanosine monophosphate (cGMP). The reduced level of cGMP results in the activation of cyclic 3′,5′‐adenosine monophosphate (cAMP)‐phosphodiesterase 3A (PDE3A), which hydrolyses cAMP. The reduced intraoocyte cAMP results in the activation of maturation promoting factor (MPF) that finally induces meiotic resumption. Thus, a transient increase of intraoocyte H₂O₂ level and decrease of NO level may signal meiotic resumption from diplotene arrest in mammalian oocytes. J. Cell. Biochem. 111: 521–528, 2010. © 2010 Wiley‐Liss, Inc.     

Reciprocal influence of connexins and apical junction proteins on their expressions and functions

Membranes of adjacent cells form intercellular junctional complexes to mechanically anchor neighbour cells (anchoring junctions), to seal the paracellular space and to prevent diffusion of integral proteins within the plasma membrane (tight junctions) and to allow cell-to-cell diffusion of small ions and molecules (gap junctions). These different types of specialised plasma membrane microdomains, sharing common adaptor molecules, particularly zonula occludens proteins, frequently present intermingled relationships where the different proteins co-assemble into macromolecular complexes and their expressions are co-ordinately regulated. Proteins forming gap junction channels (connexins, particularly) and proteins fulfilling cell attachment or forming tight junction strands mutually influence expression and functions of one another.     

Lipid droplet proteins and metabolic diseases

Lipid droplets (LDs) are ubiquitous cellular organelles for lipid storage which are composed of a neutral lipid core bounded by a protein decorated phospholipid monolayer. Although lipid storage is their most obvious function, LDs are far from inert as they participate in maintaining lipid homeostasis through lipid synthesis, metabolism, and transportation. Furthermore, they are involved in cell signaling and other molecular events closely associated with human disease such as dyslipidemia, obesity, lipodystrophy, diabetes, fatty liver, atherosclerosis, and others. The last decade has seen a great increase in the attention paid to LD biology. Regardless, many fundamental features of LD biology remain obscure. In this review, we will discuss key aspects of LD biology including their biogenesis, growth and regression. We will also summarize the current knowledge about the role LDs play in human disease, especially from the perspective of the dynamics of the associated proteins. This article is part of a Special issue entitled Cardiac adaptations to obesity, diabetes and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.