• International Journal of Industrial Entomology and Biomaterials
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• v.21 no.2
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• pp.163-167
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• 2010

[ $\alpha$ ]Glucosidase (EC 3.2.1.20) is a glycosidase that hydrolyzes disaccharides, oligosaccharides, and polysaccharides resulting in the release of α-D-glucose. In this study, $\alpha$-glucosidase activity in the hemolymph and midgut of the mulberry silkworm Bombyx mori and Japanese oak silkmoth Antheraea yamamai was measured using maltose, sucrose, trehalose, and p-nitrophenyl $\alpha$-D-glucopyranoside as substrates. In general, hemolymph $\alpha$-glucosidase activity was higher in B. mori than in A. yamamai. In contrast, midgut $\alpha$-glucosidase activity was higher in A. yamamai than in B. mori for all of the substrates tested. $\alpha$-Glucosidase activity in the midgut of both B. mori and A. yamamai showed similar responses to changes in pH and temperature for all of the substrates tested. Native (7.5%) PAGE of hemolymph and midgut proteins from B. mori and A. yamamai followed by staining with 4-methylumbelliferyl $\alpha$-D-glucoside (MUG) indicated that the $\alpha$-glucosidases of these related lepidopterans are functionally similar but structurally different. In comparison to $\alpha$-glucosidase activity from A. yamamai, $\alpha$-glucosidase activity from B. mori was generally less sensitive to the $\alpha$-glucosidase inhibitors, 1-deoxynojirimycin (DNJ), acarbose, and voglibose when the activity was determined using maltose, sucrose, and trehalose.

• Journal of Life Science
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• v.11 no.5
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• pp.415-422
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• 2001

Many secreted proteins have disulfide bonds that are important for their structure and function. Protein disulfide isomerase (PDI, EC 5.3.1.4.), an enzyme that catalyzes the formation and rearrangement of thiol/disulfide exchange reactions, is a resident of the endoplasmic reticulum (ER). The subcellular localization and its function as catalyst of disulfide bond formation in the biosynthesis of secretory and cell membrane proteins suggest that PDI plays a key role in the secretory pathway. We have isolated a cDNA encoding protein disulfide isomerase from Bombyx mori(bPDI). It has been characterized under ER stress conditions (dominantly induced by calcium ionophore A23187, tunicamycin and DTT), which is known to cause an accumulation of unfolded proteins in the ER. Furthermore, It has also been examined for tissue distribution(pronounced at the fat body), hormonal regulation (juvenile hormone, insulin and juvenile +transferrin; however, it is not effected by transferrin alone), and the effect of exogenous bacteria (peak at 16 h after infection) on the bPDI mRNA expression. The results suggest that bPDI is a member of the ER stress protein group, and it may play an important role in exogenous bacterial infection in fat body, and that homones regulate its expression.

• The Korean Journal of Zoology
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• v.26 no.2
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• pp.83-93
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• 1983

Vitellogenin was purified from the haemolymph of female pupae during egg formation of Bombyx mori using DEAE-cellulose column and its quantitative change in various organs with age ws examined by electrophoresis and immunodiffusion. Vitellogenin is distributed in the haemolymph, fat body, and over the egg maturation and especially maintainsa constant level in the haemolymph until just before emergence, indicating that vitellogenin in released into the haemolymph at the same amount as it is taken up by oocytes during pupal period. Immunologically vitellogenin was confirmed to be in the fat body until 7 days after pupation and to undergo a drastic decline thereafter. Also, the interaction of anti-ovary proteins with haemolymph proteins showed at least 3 homogeneous proteins, indicating that other proteins as well as vitellogenin are involved in egg formation.

• The Korean Journal of Zoology
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• v.37 no.1
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• pp.76-87
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• 1994

High density lipophorin-L (HDLP-L) was purified from the hemolymph of Bombyx modi using KBr density gradient ultracentrifugation and gel permeation chromatographv (Sephadex G-2001. Lipophorin has native molecular weight of 730 Nd and consists of Apo-Lp I and Apo-Lp II with molecular weights of 250 Kd and 90 Kd, respectively. Lp contains large amounts of glutamine & glutamic acid, threonine, leucine but small amounts of cysteic acid & oxidized cystine, tyrosine, methionine. Lp also contains diacylglycerol, cholesterol, phosphatidylcholine, and phosphatidylethanolamine. Anti-lipophorin showed positive reaction with fat body and ovarial extracts and also revealed immunological identity with lipophorin of Fall webworm, Hyphantria cunea. Lipophorin maintains constant level during larval and pupal stapes but greatlv increases during adult stage in both male and female. Apo-Lp III was purified from adult hemolymph. Hemolymph was subjected to KBr ultracentrifusation and Lp-free fraction was submitted to cation exchange chromatosraphy after ammonium sulfate precipitation. Apo-Lp III has molecular weight of 17 Kd, and similar amino acid composition ar those of other species Lp but contains high amounts of tryptophan which other are tacking in.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.197-201
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• 2003

The recombinant plasmids harboring a heterologous gene coding mouse endostatin were transfected and expressed stably in Trichoplusia ni Tn 5B1-4 cells and Bombyx mori BmN cells, respectively. Recombinant endostatin expressed in the stably transformed Tn 5B1-4 and BmN cells was secreted into the medium. BmN cells are relatively lower in maximum cell growth and recombinant endostatin production than Tn 5B 1-4 cells. Recombinant endostatin was also purified to homogeneity using a simple one-step ${Ni^2+}$ affinity fractionation method. Purified recombinant endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition $({ED_50})$ for recombinant endostatin was approximately 0.35 ${\mu}g$/ml.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.235-239
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• 2003

The silkworm Bombyx mori L. was studied as the potential “host” of popular in Uzbekistan biocontrol ectoparasite, entomophagous Bracon hebetor Say. Being one of representatives of economic-beneficial insects, the silkworm (larvae, pupae and imago) can be used as highly sensitive test organism for revealing of neuro toxic effects of insects venom as well as of their purified components in screening assays. In comparative aspect, except a mulberry silkworm, representatives of Uzbekistan pests cotton-boll worm Helycoverpa armigera Hbn., lesser mulberry pyralid Glyphodes pyloalis Wlk., codling moth Corpocapsa pomonella L., malaria mosquito Anopheles claviger and Colorado potato beetle Leptinotarsa decemlineata Say have been subjected to insect toxic test of bracon venom gland extract (VGE) and its fractions which were obtained by gel-chromatography on Sephadex G-100. The paralyzing effect of the VGE and its fractions was shown in a various degree on the pests.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.2
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• pp.149-152
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• 2001

Effect of topical application with 10, 20 and 30 ${\mu}g$/ml testosterone propionate and estradiol -17${\beta}$ on the fourth and fifth instar bivoltine NB18 silkworm larvae Bombyx mori, on the glycogen and protein contents of the Fat body and trehalose and protein contents of the haemolymph has been studied. Glycogen content of the fat body was significantly decreased in both testosterone propionate and estradiol -17${\beta}$ treatment groups except in the group treated with 30 ${\mu}g$ testosterone propionate where the increase was not significant when compared with those of carrier controls. The increase/decrease in haemolymph trehalose content did not show any significant difference in all the treated groups. Protein content of the fat body significantly increased in 10 and 20 mg testosterone propionate and estradiol -l7${\beta}$ treated groups but in 30 mg treated groups the increase was not significant when compared with those of carrier controls. There was no significant change in the haemolymph protein content in all the testosterone propionate and estradiol -17${\beta}$ treated groups except in group treated with 10 ${\mu}g$ estradiol -17${\beta}$ where it showed a significant decrease when compared with that of carrier control.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.2
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• pp.169-175
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• 2001

A synthetic juvenoid, R394 (Ethyl 9-cyclohexyl-3, 7-dimethyl-2, 4-nonadienoate) which is known to be a strong pest control agent was administered to silkworm, Bombyx mori L. in minute quantity for improving the silk yield. Based on the result of an earlier preliminary screening, three concentrations of the compound, viz., 0.1563, 0.3125, 31.25 nl/ml were prepared in the form of an emulsion and administered topically as a single dose, to separate batches of $5^{th}$ instar silkworm at 24, 48, 72 and 96 hrs to determine the required concentration and critical time of application for an economically favourable response. Two popular commercial silkworm hybrids, PM ${\times}$ NB4D2 (multivoltine${\times}$bivoltine) and KA${\times}$NB4D2 (bivoltine$\times$bivoltine) were subjected to the experiment. The medium and absolute control were maintained in parallel to compare the results. The results showed that 0.3125 nl/ml was the best concentration of the compound and 72 hrs of $5^{th}$instar was the most favourable age for its administration to get the maximum improvement in the commercial traits. The possible role of exogenous juvenoids in eliciting favourable response in silkworm which ultimately leads to improvement in the commercial traits is discussed.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.2
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• pp.187-190
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• 2001

Inheritance pattern of resistance to Bombyx mori nuclear polyhedrosis virus (BmNPV) was studied in an Indian silkworm stock TX by single back-cross test method. The resistant parent ［TX］, susceptible parent ［HM］, their Fl, F2, and Fl progeny back-crossed to TX [BC(R)] and HM [BC(S)] were inoculated per os with a fixed concentration of BmNPV($0.5{\times}10^{th} PIB/ml$) on the first day of second stadium. The cumulative mortality was recorded until day $10^{\times}$ post-inoculation. The results show that the resistance to BmNPV in TX fellow mono Mendelian inheritance pattern. The resistance dominated over the susceptibility at Fl. At F2, the resistant and susceptible offspring segregated in 3:1 ratio whereas at BC(S), the resistant and susceptible offspring segregated in 1:1 ratio. The response of BC(R) was more or less like the resistant parent TX which confirms the involvement of a major dominant gene conferring resistance to BmNPV in TX. The possible mechanism of inheritance of resistance in TX is discussed.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.2
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• pp.177-185
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• 2001

This study has been carried out to determine localization of Manduca sexta allatotropin (Mas-AT) neuropeptide in developing ventral nerve cord of the silk moth Bombyx mori with polyclonal antisera against Mas-AT. Suboesophageal ganglion (SOG) of the second to fifth instar larvae and 3-day-old pupae showed two to ten Mas-AT-immunoreactive (Mas-AT-IR) cell bodies. There were two to three pairs of labeled cell bodies in each thoracic ganglion (TG) from third instar larvae to adults, with the exception of TG from prepupae. One pair of labeled cell bodies was localized in each abdominal ganglion (AG) 1 to 6 from third instar larvae to 3-day-old pupae, whereas in 5-day-old pupae to adults there was one pair in a similar location of AG 1 to 5. The seventh neuromeres of terminal abdominal ganglia (TAG) from third instar Iarvae to 3- day-old pupae contained four labeled large cell bodies. In each of AG 1 to 7, these cell bodies showed similar allatotropin-immunoreactivity in appearance. Some labeled axons, projected from Mas-AT-lR cells in each of those AG, were extended to the nerves N 1 and N 2.