Peptides in the hemolymph of Hermetia illucens larvae completely inhibit the growth of Klebsiella pneumonia in vitro and in vivo

Extensive use of antibiotics has caused the microbial resistance to rise drastically within the last few decades, and new approaches are therefore needed to develop effective antibacterial substances. In this study, we identified peptide in the hemolymph of Hermetia illucens larvae using reverse-phase chromatography, HPLC and Nano-LC-ESI-MS/MS system. We investigated the antibacterial effect of HP/F9 peptides against Klebsiella pneumoniae in vitro and in vivo. The peptide effectively inhibited the growth of K. pneumoniae in vitro and completely removed K. pneumoniae from the lungs of mice. Importantly, peptides (22,000 Da, HP/F9) successfully reduced lung inflammation upon K. pneumoniae infection. These results indicate that the HP/F9 peptide from H. illucens larva can effectively protect the mouse from K. pneumoniae infection. HP/F9 could be a new candidate for the development of effective antibacterial substance.     

Characteristics of insecticidal substances from the entomopathogenic fungus Metarhizium pinghaense 15R against cotton aphid in Korea

Aphids (Hemiptera: Aphididae) are agricultural pests for a wide range of economically important crops worldwide. Entomopathogenic fungi have been studied and developed as a biological agents to control hard to control insect pests like the aphid. In particular, secondary metabolites produced by entomopathogenic fungi are being studied as insecticidal substances. This study evaluated the insecticidal activity and characteristics of insecticidal substances in culture filtrate against aphids to confirm the insecticidal substances produced by Metarhizium pinghaense 15R. After treating cotton aphids with the culture filtrate, strong insecticidal activity (100 % insect mortality) was observed on the third day of treatment, and it was confirmed that insecticidal substances in the culture filtrate had high thermal stability. In addition, the physicochemical properties of insecticidal substances in the culture filtrate were evaluated, and it was determined that the insecticidal substances were hydrophobic, with small molecules of 3 kDa or less. Based on these results, it was suspected that the insecticidal substances in the culture filtrate were destruxins (DTXs), which are representative secondary metabolites of Metarhizium known to have insecticidal activity. Through HPLC (High Performance Liquid Chromatography) analysis, the presence of DTX A and B was found in all active samples, and it was confirmed that the insecticidal substances of M. pinghaense 15R were DTXs. This study provides primary data for developing biochemical crop protection agents using these insecticidal substances, and we suggest the potential of using insecticidal substances of entomopathogenic fungi as biochemical pesticides in Korea.     

Bioactive substances produced by marine isolates of Pseudomonas

Pseudomonas is a genus of non-fermentative gram-negative Gammaproteobacteria found both on land and in the water. Many terrestrial isolates of this genus have been studied extensively. While many produce bioactive substances, enzymes, and biosurfactants, other Pseudomonas isolates are used for biological control of plant diseases and bioremediation. In contrast, only a few marine isolates of this genus have been described that produce novel bioactive substances. The chemical structures of the bioactive substances from marine Pseudomonas are diverse, including pyroles, pseudopeptide pyrrolidinedione, phloroglucinol, phenazine, benzaldehyde, quinoline, quinolone, phenanthren, phthalate, andrimid, moiramides, zafrin and bushrin. Some of these bioactive compounds are antimicrobial agents, and dibutyl phthalate and di-(2-ethylhexyl) phthalate have been reported to be cathepsin B inhibitors. In addition to being heterogeneous in terms of their structures, the antibacterial substances produced by Pseudomonas also have diverse mechanisms of action: some affect the bacterial cell membrane, causing bacterial cell lysis, whereas others act as acetyl-CoA carboxylase and nitrous oxide synthesis inhibitors. Marine Pseudomonas spp. have been isolated from a wide range of marine environments and are a potential untapped source for medically relevant bioactive substances.     

Induction of oviposition by injection of male-derived extracts in two Callosobruchus species

In some insect species, certain substances in the seminal fluid of males induce egg production and laying in females. We determined the effects of male-derived substances on female oviposition behaviour in two Callosobruchus species, C. chinensis and C. maculatus. Aqueous extracts of the accessory gland; testis; and seminal vesicle, including the ejaculatory duct, were prepared. The injection of these extracts into abdomen of females induced oviposition in both species. Oviposition was induced by the testis and seminal vesicle extracts in C. chinensis and by the accessory gland extracts in C. maculatus. The extracts were separated into three fractions by ultrafiltration: fractions I, molecular weight (MW) <3 kDa; fraction II, 3-14 kDa; and fraction III, >14 kDa. Fraction III induced oviposition in both species. These results suggest that in these two species, the substances that induce oviposition have similar MW but are present in different organs. Oviposition was induced by high-MW (>14 kDa) substances in the testis and seminal vesicle in C. chinensis, and by high-MW substances in accessory gland in C. maculatus. Here, we have discussed the relationship between oviposition and the abovementioned male-derived substances.     

In vitro diapause substance in the silkworm, Bombyx mori

By means of in vitro studies, in which isolated suboesophageal ganglions of the Bombyx silkworm were cultured, it was shown that at least two kinds of substances are biosynthesized and exert independent effects on determination of diapause or non-diapause in silkworm eggs. They are referred to as the diapause and non-diapause substance, respectively. Whether diapause or non-diapause eggs are laid may depend upon the different quality of these substances.     

Cardopatine and isocardopatine,two novel cyclobutane substances from Cardopatium corymbosum

Two new natural substances containing a cyclobutane unit, cardopatine and isocardopatine, the trans and cis isomers respectively of 5,5″ (cyclobut-1,2-ylene-diethynylene)bis 2,2′-bithiophene), together with the known α-terthienyl and 5-(but-3-en-1-ynyl)-2,2′-bithienyl, have been isolated from the roots of Cardopatium corymbosum. Evidence is given that the novel cyclobutane substances are not the products of a spontaneous dimerization of a bithienyl monomeric unit. Structure determination and conformational analysis are reported.     

Role of Humic-Bound Iron as an Electron Transfer Agent in Dissimilatory Fe(III) Reduction

The dissimilatory Fe(III) reducer Geobacter metallireducens reduced Fe(III) bound in humic substances, but the concentrations of Fe(III) in a wide range of highly purified humic substances were too low to account for a significant portion of the electron-accepting capacities of the humic substances. Furthermore, once reduced, the iron in humic substances could not transfer electrons to Fe(III) oxide. These results suggest that other electron-accepting moieties in humic substances, such as quinones, are the important electron-accepting and shuttling agents under Fe(III)-reducing conditions.     

Lichen substance concentrations in the lichen Hypogymnia physodes are correlated with heavy metal concentrations in the substratum

Lichen substances (i.e. lichen-specific carbon-based secondary compounds) are known to be involved in the uptake and immobilization of metal ions, though the biochemical mechanisms of this interaction are largely unexplained. Previous research on potential effects of lichen substances on heavy metal uptake and tolerance mostly focused on lichens in heavily polluted areas with exceptionally high metal concentrations. In the present study, we aimed at gathering information as to whether lichen substances might be involved in the fine-tuning of metal uptake even at not or low-polluted sites. Therefore, we studied lichen substance concentrations in the epiphytic lichen Hypogymnia physodes and metal concentrations in its substratum in a montane spruce forest of Germany. H. physodes produces two depsides and five depsidones, which had been shown to be involved in metal homeostasis, namely in Cu and Mn uptake, in previous laboratory experiments. The amount of lichen substances increased with increasing heavy metal concentration in the substratum, though the latter varied only in the range of a few μmol g⁻¹ between the sample trees. Variability of lichen substance concentrations in H. physodes within the individual trees was low. Among the different lichen substances of H. physodes, the amount of the depsidone physodalic acid relative to the total of lichen substances was most closely correlated to the concentrations of Cu and Mn in the substratum, whereas the amount of the depsidone 3-hydroxyphysodic acid decreased both with increasing concentrations of these two metals and physodalic acid. Thus, our data suggest that lichen substances contribute to metal homeostasis not only in heavy metal-rich habitats, but also at not or low-polluted sites where the lichen substances apparently help to maintain constant intracellular metal concentrations despite of spatially varying availabilities of metal ions.     

两种水溶性抗菌活性物质的分离提取

对水溶性、不解离的极性物质分离时,一般采用吸附层析和凝胶层析等途径。实验通过硅胶柱层析、葡聚糖凝胶柱层析以及硅胶ＧＦ２５４制备型薄板层析,从发酵液样品中分离出两种有抗菌活性的纯物质。薄层层析的展开剂为二氯甲烷－四氢呋喃－甲醇－水（２５：３０：２）,分离出的组分中Ｒｆ＝０．７和Ｒｆ＝０．８两种物质有抗菌活性。硅胶柱层析洗脱过程为梯度洗脱,先用１５０ｍｌ上述展开剂洗脱,再用二氯甲烷－甲醇（２０：８０）     

Affinity chromatography of neoglycoproteins

Glycoproteins, as a class of biomolecules, exhibit much more heterogeneous structures than non-glycosylated proteins. They present a challenging area of research. Model glycoproteins with well-defined protein and carbohydrate structures are helpful in the search for high-resolution methods for the separation of glycoproteins. Neoglycoproteins, maltose-modified chymotrypsin and lactose-modified chymotrypsin, were synthesised by modifying chymotrypsin with maltose and lactose, respectively, using the reductive amination method. Boronate chromatography was applied to isolate the neoglycoproteins from non-glycosylated substances. The use of Tris–HCl as a shielding reagent during the boronate chromatography proved to be efficient in eliminating unwanted interactions between the boronate ligand and the peptide backbone of chymotrypsin. The retention time of neoglycoproteins on the boronate column was increased with increasing the degree of modification.     

Pheromones released during inter- and intra-sex response of the scolytid beetle Dendroctonus brevicomis

Pheromones of Dendroctonus brevicomis released variously during inter- and intra-sex response (including stridulation by both sexes) were the known pheromones exo-brevicomin, endo-brevicomin, frontalin and verbenone, and substances identified as pinocarvone, trans-pinocarveol, and myrtenol. These substances are present in emergent beetles and thus attack of a host tree is not essential for their initial production.     

Effect of platelet activating factor on leukocytes II. Enhancement of eosinophil chemotactic factor and β-glucuronidase release

Synthetic 1-O-alkyl-2-O-acetyl-sn-glyceryl-3-phosphorylcholine (PAF) and 1-O-alkyl-sn-glyceryl-3-phosphorylcholine (lyso-PAF) have previously been shown to induce chemotaxis and chemokinesis of human neutrophils. We present here data showing that these agents are inactive by themselves, but that they enhance neutrophil secretion once it has been initiated by a calcium ionosphore or by zymosan. Two substances, the lipid eosinophil chemotactic factor (ECF) and the lysosomal enzyme β-glucuronidase, are used as markers for neutrophil release. PAF augments secretion of both substances in a dose-dependent fashion, with lyso-PAF being less potent. The kinetics of enhancement are very rapid (<2 min) and are not reversible by washing of the cells. A pyrazoline derivative that inhibits arachidonate cyclo-oxygenation and lipoxygenation, reduces the enhancing effect of PAF and lyso-PAF. PAF, and less so lyso-PAF, are thus potentially important modulators of neutrophil secretion during inflammatory processes.     

Phenylhydrazides as inhibitors of Leishmania amazonensis arginase and antileishmanial activity

Searching for new substances with antileishmanial activity, we synthesized and evaluated a series of α,α-difluorohydrazide and α,α-difluoramides against Leishmania amazonensis arginase (LaArg). Four α,α-difluorohydrazide derivatives showed activity against LaArg with K_i in the range of 1.3–26 μM. The study of the kinetics of LaArg inhibition showed that these substances might act via different inhibitory mechanisms or even by a combination of these. The compounds were tested against L. amazonensis promastigotes and the best result was obtained to the compound 4 (EC₅₀ of 12.7 ± 0.3 μM). In addition, in order to obtain further insight into the binding mode of such compounds, molecular docking studies were performed to obtain additional validation of experimental results. Considering these results, it is possible to conclude that α,α-difluorohydrazide derivatives are a promising scaffold in the development of new substances against the etiological agent of leishmaniasis by targeting LaArg.     

Microstructure of the prothoracic mycangia in the ambrosia beetle Platypus koryoensis (Coleoptera: Curculionidae: Platypodinae)

Platypus koryoensis (Murayama) (Coleoptera: Curculionidae: Platypodinae) is a minute ambrosia beetle that can cause significant damage to live oak trees (Quercus mongolica) along with its symbiotic pathogenic fungus (Raffaelea sp.). Although both sexes of the beetle have a number of cuticular depressions for carrying microorganisms, only the females have mycangia that are typical in size on the pronotum. The mycangial cavities are filled with a mass of amorphous substances including yeast-like spores. In addition, mycangial cavities are equipped with cuticular pores which release substances to support symbiotic microorganisms during storage. Small depressions of both sexes can be subdivided into two clusters according to their relative sizes and locations. Although they have a typical cuticular peg (basiconic sensillum), they also share a common morphological characteristic with the typical mycangium of female: a round outlet and an invaginated reservoir containing a mass of secretory substances. Accordingly, our fine structural data suggest that both the female mycangia and the clusters of small cuticular depressions in both sexes also have a mycangial function in common regardless of their relative size and location.     

Screening of poplar trees for antibacterial,antifungal and antiviral activity

The leaves of poplar trees (genusPopulus) contain antimiorobial agents. These antibiotic properties are markedly influenced by interspecific and intraspecific hybridizations. Fractionation of the weakly active crude extracts results in more active antibacterial fractions. There are indications that the substances which inhibit antimicrobial effects during the testing procedure remain in the methanol fractions. In addition the results show that the antibacterial agents differ from the antifungal compounds, which both are different from the antiviral substances.     

Peach Leaf Senescence Delayed by Criconemella xenoplax

Fall annual leaf senescence of peach was delayed in the field and in microplots in the presence of Criconemella xenoplax. Soil from the rhizosphere of orchard trees with greener leaves had ca. 2.5 × more nematodes than soil around trees in a more advanced state of fall senescence. In microplots, monoclonal antibody enzyme immunoassay (EIA) of leaf cytokinins indicated that concentration of zeatin riboside-like substances and chlorophyll content were greater in leaves of trees growing in nematode-infested soil than in trees in uninfested soil. EIA also indicated the presence of substances resembling trans-zeatin, zeatin riboside, dihydrozeatin, and dihydrozeatin riboside-like substances in whole body homogenates of C. xenoplax. Levels of zeatin-like substances were present in the nematode in greater levels than the other related substances.     

The Effects of Plant Growth Substances and Mixed Cultures on Growth and Metabolite Production of Green Algae Chlorella sp.: A Review

Recent interest in the use of microalgae for the production of biofuels and bioproducts has stimulated an interest in methods to enhance the growth rate of microalgae. This review examines past work involving the stimulation of Chlorella sp. growth and metabolite production by plant growth substances as well as by mixed cultures of Chlorella sp. with bacteria. Plant growth substances known to regulate Chlorella sp. growth and metabolite production include auxins, cytokinins, abscisic acid, polyamines, brassinosteroids, jasmonic acid, salicylic acid, and combinations of two or three of the aforementioned substances. Mixed cultures of bacteria are examined, including both natural bacteria–algae consortia and artificially induced symbioses. For natural consortia, commonly occurring bacterial species, including the genera Brevundimonas and Sphingomonas, are discussed. For artificially induced symbioses, the use of the nitrogen-fixing bacterium Azospirillum is examined in detail. In particular, a variety of studies have involved the coimmobilization of Chlorella sp. with Azospirillum sp. in alginate beads, with the goal of using the mixed culture to treat wastewater. In summary, the use of plant growth substances and mixed cultures provides two methods to increase the growth of Chlorella sp., whether for the production of lipids for biofuels, the production of bioproducts, the treatment of wastewater, or a variety of other reasons.     

牛膝根结构特征与三萜皂苷的积累关系

三萜皂苷是牛膝根中主要的药用成分之一。利用皂苷定量分析与解剖学和组织化学定位方法对不同生长年限牛膝根的解剖结构及三萜皂苷的积累动态关系进行了研究。结果表明：牛膝根属于异常结构,三生结构发达。牛膝根的增粗生长主要依赖于额外形成层细胞的分裂活动,牛膝根中最多可以产生7～8圈额外形成层。次生韧皮部、栓内层和三生韧皮部是三萜皂苷的主要积累场所;三萜皂苷的积累动态随牛膝的生长年限增加呈现低→高的变化规律,3～4年生牛膝跟中三萜皂苷的积累达最大值。     

Toxic-Metabolite-Producing Bacteria and Fungus in an Indoor Environment

Toxic-metabolite-emitting microbes were isolated from the indoor environment of a building where the occupant was suffering serious building-related ill-health symptoms. Toxic substances soluble in methanol and inhibitory to spermatozoa at <10 μg (dry weight) ml⁻¹ were found from six bacterial isolates and one fungus. The substances from isolates of Bacillus simplex and from isolates belonging to the actinobacterial genera Streptomyces and Nocardiopsis were mitochondriotoxic. These substances dissipated the mitochondrial membrane potential (Δψ) of boar spermatozoa. The substances from the Streptomyces isolates also swelled the mitochondria. The substances from isolates of Trichoderma harzianum Rifai and Bacillus pumilus damaged the cell membrane barrier function of sperm cells.     

Micronuclei in red blood cells of the newt Pleurodeles waltl Michah: induction with X-rays and chemicals

Pleurodeles waltl, a typical long-tailed amphibian (Urodela) is proposed as a new animal for genetic toxicological studies. X-Rays and various clastogenic substances cause the formation of clearly visible micronuclei in the red blood cells (RBC). The proportion of cells with micronuclei was determined from blood smears of larvae after irradiation or after having been kept in water containing the substances to be studied. A dose-effect curve was established for X-irradiation. The 6 following substances were tested by this method: benzo[a]pyrene, carbaryl, N-nitrosocarbaryl, aziridine, caffeine and formaldehyde. Formaldehyde was the only substance tested that did not bring about formation of micronuclei in the RBCs. The results were compared with data already obtained by other methods of toxicology.This method should allow a cytogenetic test to be developed for the detection of clastogenic substances in aqueous media.