Immune-Enhancing Activity Screening on Extracts from Two Crickets, Gryllus bimaculatus and Teleogryllus emma

This study was performed to explore novel and valuable uses of insect resources, important subjects of the natural compound used in bio‐industries. The whole bodies of two crickets, Gryllus bimaculatus and Teleogryllus emma, selected from medicinal insect species, were carefully ground and treated with 80% EtOH. The insect extracts were solubilized and separated by hexane, butanol, and D.W according to their polarities. Three types of extracts, a D.W fraction (G1) and a boiling extract (G2) of an introduced cricket, G. bimaculatus, and a D.W fraction (T1) of a Korean local cricket, T. emma, were prepared to assay immune stimulating activity of cricket originated compounds. The all of three treated cricket extracts showed to increase IL‐4, IFN‐, and TNF‐α. Among those extract, extract G2, boiled extract from G. bimaculatus, was the best immune–enhancing fraction. The results of this study could be fundamental information for further works to use insects as natural resources having plenty of potentials and varieties.     

Leaf Anatomy and Carbon Discrimination in NAD-malic Enzyme Panicum Species and their Hybrids Differing in Bundle Sheath Cell Ultrastructure

The relationship between leaf anatomy, ultrastructure and carbondiscrimination was investigated in leaves of two F₁hybrids (F₁-1and F₁-2) between two different types of the grassPanicum [anNAD-malic enzyme (ME) C₄species], which differ in bundle sheathultrastructure. The female parent was Kabulabula grass, whichhas centrifugal chloroplasts in bundle sheath cells and is designatedan NAD-ME(F) species, while the male parent was Makarikari grass,which has centripetal chloroplasts in the bundle sheath cellsand is designated an NAD-ME(P) species. Suberin lamellae arepresent in Kabulabula grass but are lacking in Makarikari grass.Both F₁hybrids had the same chromosome number (2n =36) as theparents but exhibited both univalent (about 45%) and bivalent(about 55%) chromosome pairing which was the major basis forthe identification of F₁hybrids. In F₁-1, elongated bundle sheathcell chloroplasts are arranged mainly in a centripetal position,similar to those in the male parent, Makarikari grass. In contrast,most of the bundle sheath cells in F₁-2 are packed with starch-containingchloroplasts, although in some cells chloroplasts tended tobe centripetally arranged. In both F₁hybrids, suberin lamellaewere found in the bundle sheath cell walls, similar to the femaleparent, Kabulabula grass. The ¹³C values of both F₁hybrids were-11.4 to -11.7, almost the same as those of Kabulabula grass(-11.4), but significantly higher than those of Makarikari grass(-12.7). These results indicate that the chloroplast orientationin the bundle sheath cells and the presence of suberin lamellaeare not obligatorily linked in their expression and suggestthat suberin lamellae may play an important role in discriminationagainst¹³C. Panicum ; NAD-malic enzyme species; hybrid; chloroplast position; ¹³C discrimination; suberin lamellae     

Behavioural and chemical mechanisms of plant‐mediated deterrence and attraction among frugivorous insects

1. Herbivory often induces systemic plant responses that affect the host choice of subsequent herbivores, either deterring or attracting them, with implications for the performance of both herbivore and host plant. Combining measures of herbivore movement and consumption can efficiently provide insights into the induced plant responses that are most important for determining choice behaviour. 2. The preferences of two frugivorous stink bug species, Nezara viridula and Euschistus servus between cotton plants left undamaged or damaged by Helicoverpa zea and Heliothis virescens larvae were investigated. A novel consumer movement model was used to investigate if attraction rates or leaving rates determined preferences. Stink bug consumption rates were measured using salivary sheath flanges. Finally, the systemic induction of selected phenolic‐based and terpenoid secondary metabolites were measured from heliothine herbivory on developing cotton bolls, to investigate if they explained stink bug feeding responses. 3. Heliothine herbivory did not affect the N. viridula feeding preference. However, we found opposing effects of H. zea and H. virescens herbivory on the behaviour of E. servus. Avoidance of H. zea‐damaged plants is not obviously related to phenolic or terpenoid induction in cotton bolls; whereas a preference for H. virescens‐damaged plants may be related to reductions in chlorogenic acid in boll carpel walls. 4. The present results highlight the inferential power of measuring both consumer movement and consumption in preference experiments and combining behavioural responses with phytochemical responses. Furthermore, while plant‐mediated interactions among herbivorous insects are well studied, interactions among frugivorous species specifically have been poorly documented.     

Establishment of a novel dwarf rat strain: cartilage calcification insufficient (CCI) rats

Rats with dwarfism accompanied by skeletal abnormalities, such as shortness of the limbs, tail, and body (dwarf rats), emerged in a Jcl-derived Sprague-Dawley rat colony maintained at the Institute for Animal Experimentation, St. Marianna University Graduate School of Medicine. Since the dwarfism was assumed to be due to a genetic mutation based on its frequency, we bred the dwarf rats and investigated their characteristics in order to identify the causative factors of their phenotypes and whether they could be used as a human disease model. One male and female that produced dwarf progeny were selected, and reproduction was initiated by mating the pair. The incidence of dwarfism was 25.8% among the resultant litter, and dwarfism occurred in both genders, suggesting that it was inherited in an autosomal recessive manner. At 12 weeks of age, the body weights of the male and female dwarf rats were 40% and 57% of those of the normal rats, respectively. In soft X-ray radiographic and histological examinations, shortening and hypoplasia of the long bones, such as the tibia and femur, were observed, which were suggestive of endochondral ossification abnormalities. An immunohistochemical examination detected an aggrecan synthesis disorder, which might have led to delayed calcification and increased growth plate thickening in the dwarf rats. We hypothesized that the principal characteristics of the dwarf rats were systemically induced by insufficient cartilage calcification in their long bones; thus, we named them cartilage calcification insufficient (CCI) rats.     

A New Species of the Genus Amblyseius from Korea (Acari: Phytoseiidae)

A new species, Amblyseius choii sp. nov. is described from Korea. The new species was collected on the deciduous plants, Acer pseudo‐sieboldianum (Paxton) Kom. at Mahyeon‐ri, Geunnam‐myeon, Cheorwon‐gun, Gangwon‐do Province.     

Change of Haemolymph Ferritin of Sweet Potato Hornworm, Agrius convolvuli by Heavy Metals

Agrius convolvuli haemolymph ferritin was purified by KBr density gradient ultracentrifugation and anion exchange column chromatography. The 670 kDa ferritin was composed of two subunits of 26 kDa and 31 kDa. It was also shown that the protein had an isoelectric point (pI) of pH 7.4. The N‐terminal amino acid sequences of the two subunits were NH2‐DNXYQDVSLDXSQAXNXL (26 kDa subunit) and NH2‐TQXHVNPVNIQRDXVTMHXS (31 kDa subunit). The sequential analysis showed that they had high similarity to lepidopteran ferritin subunits, S‐ and G‐type, respectively. Using electron microscope, it was observed that the protein had a core whose size was about 7 nm. In the amino acid composition of the protein, Glu (13.22%), Asp (10.43%), Pro (9.69%), Leu (9.63%), Ala (9.55%) and Gly (8.49%) were in relatively high contents while Tyr (1.21%), His (2.58%) and Arg (3.10 %) were in low. It was shown that the amount of ferritin in A. convolvuli haemolymph was increased by injection of eight different heavy metal ions, FeCl₃, HgCl₂, CuSO₄, ZnSO₄, MnCl₂, MgCl₂, CrCl₃ and CdCl₂. Among the ions, Fe³⁺, Hg²⁺, Zn⁴⁺, Mn²⁺ and Cd.²⁺ significantly induced the amount of the protein.     

Rapid and Efficient Purification of Haemolymph Ferritin from Cricket, Gryllus bimaculatus

The Gryllus bimaculatus ferritin was purified from the haemolymph by a consecutive four‐step procedures consisting of 50% ammonium sulfate fractionation, anion exchange column chromatography using HiTrap^TM Q column (1.6 x 4 cm, Amersham Bioscience), 70°C heat treatment for 10 min, acid treatment of 0.1 M sodium acetate buffer (pH 6.0), and gel filtration column chromatography using G4000SW column (0.75 x 60 cm, Tosoh, Japan) connected on FPLC system. The purified ferritin was found to have two major subunits of 32 and 30 kDa and three minor subunits of 28, 27, and 25 kDa by 2D electrophoresis analysis. Amino acid composition analysis showed that there are high contents of Asp, Glu, Met, Leu, and Lys residues in ferritin while low contents of Cys, Tyr, and Trp residues in the protein. G. bimaculatus haemolymph ferritin could be classified as a methionine‐rich protein.