• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.2
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• pp.127-131
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• 2002

The qualitative and quantitative changes of bacterial flora associated with the Pure Mysore (Multivoltine) and NB$_4$D$_2$ (Bivoltine) silkworm (Bombyx modi L.) midgut during third, fourth and fifth instars were studied. Larvae reared on mulberry leaves were dissected and their midgut bacterial populations were enumerated through serial dilution technique and after 72 hrs of incubation period at 28 $\pm$ 1$^{\circ}C$, the bacterial population was estimated. The results showed a highest mean value of 15$\times$10/ sup 6/ sup 6/ CFU/g and 28$\times$10/ sup 6/ CFU/g in Pure Mysore and NB$_4$D$_2$races, respectively, in midgut tissue of fifth instar larvae. The natural epiphytic microflora of mulberry leaves fed during the respective instars was also studied and found maximum 14$\times$10$^3$ CFU/g in leaves fed in third instars, followed by 5.3$\times$10$^3$ CFU/g and 2.1$\times$10$^3$ CFU/g in leaves fed during fourth and fifth instars, respectively. The bacterial flora colonized in midgut was found to be elaborating amylase, caseinase, gelatinase, lipase and urease enzymes. The highest percentages of isolates were amylase producers followed by protein and lipid splitters in Pure Mysore, whereas in NB$_4$D$_2$ protein splitter were dominated followed by lipase and amylase producers in NB$_4$D$_2$. The results indicate that the natural microflora may play a vital role in the digestion of ingested food materials in silkworms.

• Journal of Sericultural and Entomological Science
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• v.2
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• pp.49-61
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• 1962

The insect Diplosis mori Yokoyama is causing extensive destruction of mulberry trees in Korea with a resultant loss in silk production. This study was made to determine an effective method of control. Methods and Materials Used Preliminary studies were made to determine more exactly the life cycle of the insect. Based on this information, various control measures were tested, including the use of spray methods with BHC and control of larvae by tilling. Results Obtained 1. Life cycle studies (a) In the Suwon area, this-insect has 5 generations per year. The first starts in the later part of June and the final cycle ends in the later part of September. (b) The adult insects appear about 7: 00-8: 00 P.M. and live for 2-5 days. Females live in longer periods than the male. (c) Larvae lives inside the second and third stipules (A. B.) before mulberry leaf development. They cause extensive damage to the leaves at the point where they are attached to the stem. (d) Weather conditions considerably affect the life cycle. The pupa particularly are affected and not be able to change into the moth stage when there is a long period of no rain. (e) Larvae are large......0.3 to 2.0mm......and are milky-white immediately after hatching but turn to pinkish as the worm matures. The matured worm has a jumping ability up to 15-20cm. The worm burrows into the ground 1.5 to 3.0 cm before changing into the pupal stage. (f) The pupal stage usually lasts 7-8 days, in summer weather conditions and the pupa is surrounded with a coarse cocoon. (g) These insects, as a general rule, overwinter as pupae but sometimes as larvae. 2. Control measures (a) BHC dust applied on the ground seem most effective. It should be done 4-5 days after the worm has burrowed into the ground. For this control, it is recommended that 6kg of a 2% formation Tanbo(l0ares) be used. (b) For the effective spraying against the fly, it is recommended that a formulation of liquid BHC spray terials be used at the rate of 400-600 liters per Tanbo. (c) Tillage methods which provide a cover of soil 5cm or more in depth above infested areas will effect-maively prevent the emergence of the fly from the pupal stage. 3. Conclusions Methods of control against Diplosis mori Yokoyama can be tied more closely to the life cycle of the insect with more effective results. Further studies are needed to complete information on possible controls during or after hibernation. Economic studies on the cost of these control measures are also needed.

• Korean journal of applied entomology
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• v.36 no.4
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• pp.331-340
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• 1997

Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.18-24
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• 2001

Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

• Molecules and Cells
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• v.25 no.4
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• pp.467-478
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• 2008

Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

• Journal of Sericultural and Entomological Science
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• v.46 no.2
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• pp.77-79
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• 2004

To investigate whether BF-7, extracted from Bombyx mori, improved learning and memory of ordinary people, K-WAIS (Korean version of Wechsler adult intelligence scale) was performed in 4 normal students. Treatment with 400 mg of BF-7 increased mean IQ from 103 to 114. To know how BF-7 plays such a positive role, we measured the blood flow to brain, especially for the area concerned with learning and memory, with Single Photon Emission Computed Tomography(SPECT). Our result showed that the blood flow to parahippocampal gyrus and medial temporal area was increased. Also, our results showed the image representing the increase of blood supply in this area. So, our results suggest that BF-7 effectively help to use brain concerning with learning and memory.

• Journal of Acupuncture Research
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• v.23 no.6
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• pp.29-44
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• 2006

Objectives : The purpose of this study is to observe the effects of Mori Ramulus herbal-acupuncture solution(MR-HAS) on arthritis of mice induced by Collagen II at Joksamni(ST36). Methods : The author performed several experimental items. First, it is the cell survival rate of mice lung fibroblasts. Second, it is the incidence rate of arthritis and arthritis index of CIA. Third, it is the levels of IL-6, $TNF-{\alpha}$, $IFN-{\gamma}$, $IL-1{\beta}$, IgG, IgM and anti-collagen II in serum and the level of $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$ ratio in CIA mouse spleen cell culture. Fourth, it is histological analysis of the mice joint. Fifth, it is expression ratio of CD3e+ to CD19+ cell, CD4+ to CD8+ cell, CD69+/CD3e+ cells, CD11a+/CD19+ cells and CD11b+/Gr-l+ cells and CD4+/CD25+ cells. Results : 1. In the MR-HA, the incidence of arthritis and the arthritis index were significantly decreased. 2. In MR-HA, the levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$, IgG, IgM and anti-collagen II in serum of CIA mice and the level of $IFN-{\gamma}$, IL-4, $IFN-{\gamma}$, IL-4 ratio in CIA mouse spleen cell culture were significantly decreased. 3. In histology, the cartilage destruction and synovial cell proliferation were decreased in the MR-HA, and the collagen fiber expressions in the MR-HA were similar with that of the Normal group. 4. In the MR-HA, the expression ratio of CD3e+ to CD19+ cell and CD4+ to CD8+ cell were similarly maintained as Normal group in lymph nodes, and CD69+/CD3e+ cells and CD11a+/CD19+ cells were decreased in lymph nodes, and CD11b+/Gr-1+ cells and CD4+/CD25+ cells were decreased in synovium. Conclusion : These results suggest that MR-HA at ST36 has an effect to control synovial cell proliferation and cartilage destruction in rheumatoid arthritis, as well as prophylaxis is important to treat rheumatoid arthritis in clinic.

• Journal of Sericultural and Entomological Science
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• v.43 no.2
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• pp.59-66
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• 2001

This study was conducted to elucidate phylogenetic relationships and genetic characterization of silkworms that might be recognized as the Korean native strains. Genetic characterization in isozymes and the proteins of larval hemolymph of 17 silkworms were observed by acrylamide gel eletrophoresis, on 12 genes; Bph, Bes, les, Amy-hc, Ict-A, -B, -D,-E,-H, Pfl, Pst, Lp. Gene frequencies in each locus were compared other geographic strains. Korean native strains were remarkably different from others considered as the genetic characterization of Korean native strains. Phylogenetic relationships in Korean native strains were analysed using RAPD-PCR markers. A total of 40 primers were used and 346 bands of amplified DNA were generated from geographic strains. Genetic similarity based on the RAPD bands was used to construct phylogenetic dendrogram based on analysis of bard sharing data of amplified markers. Genetic similarity ranged from 0.595 to 0.860. In the genetic relationship based on dendrogram, they were classified into Bombyx mori group (including 16 domesticated silkworm strains) and B. mandarina group. The Bombyx mori group was separated into three sub-groups at the genetic similarity of 0.6930, including Korean, Japanese and Chinese groups. According to this result, the Korean native variety can be considered as a clearly different variety from other geographic strains. It may be concluded that the Korean native strains are also one of original geographic variety such as Japanese, Chinese, etc.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.69-73
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• 2001

The effect of topical application 100,300 and 500 $\mu$g/ ml with Mineral Salts of magnesium sulphate and potassium nitrate on economic parameters was ana- lyzed following treatment of last larval stadium. The treated larvae showed significantly decreased larval weight and silk gland weight in all the treated groups along with other decreased Iarval, cocoon and adult parameters. The female cocoon weight was significantly decreased in all the treated groups with decreased female cocoon shell weight and male cocoon weight at 300 and 500$\mu$g/ml respectively. The length and weight of filament was significantly decreased at 300 and 500 $\mu$g/ml respectively and denier at 500 $\mu$g/ ml. The fecundity decreased significantly in 300 and 500 $\mu$g/ml treated groups when compared with the corresponding parameters of the carrier control.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.93-96
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• 2001

The effect of supplementation with potassium nitrate on biochemical constituents was analyzed following treatment of last larval stadium. The fat body glycogen and haemolymph trehalosre significantly increased in all the treated groups. The fat body and haemolymph protein also increased significantly in all the treated groups except 500 ${\mu}g$/ml where increased fat body protein and decreased haemolymph protein were not significant. The total lipids, phospholipids and neutral lipids of the fat body increased significantly in all the groups except 50 and 300 ${\mu}g$/ml where the fat body phospholipids increased but the increase was not significant. However the total lipids, phospholipids and neutral lipids are significantly decreased in 500 ${\mu}g$/ml treated group when compared with that of the carrier control.