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M Sumida

Publications and source records attributed to M Sumida.

157 records · Page 9Linked to original sources

Antibacterial activity inducible in the haemolymph of the silkworm, Bombyx mori, by injection of formalin-treated Escherichia coli K-12 during the fifth larval instar and pharate adult development.

1. Antibacterial activity inducible in the haemolymph of the silkworm, Bombyx mori, by immunization, i.e. by injection of formalin-treated Escherichia coli (E. coli) K-12 during the fifth larval instar and pharate adult development that was reared aseptically on an artificial diet was determined by inhibition zone assay using the same bacterium as a test organism. 2. A peak of antibacterial activity was observed in each development stage; approximately 8 mm in diameter of a clear zone at days 3 or 4 in the fifth larval instar and approximately 5 mm at day 1 in the pharate adults. 3. Acid polyacrylamide gel electrophoresis of immunized haemolymph followed by overlay assay showed that an activity band was associated with two peptide bands that were similar to the cecropin-like peptides A and B that were reported in the silkworm (Morishima et al., 1988, Agri. Biol. Chem. 52, 929-934). Any other activity bands were not observed. No activity band was detectable from the haemolymph of non-immunized insects. 4. Fractionation of antibacterial peptides in immunized haemolymph on a CM-cellulose column resulted in separation of two groups of activity, both in the fifth instar larvae and in the pharate adults with a slight difference in elution conditions. 5. Duration of high antibacterial activity induced by a single immunization was approximately 12 hr in the fifth instar day 3 larvae and 48 hr in the day 2 pharate adults.

Animals↗

Induction of antibacterial activity in the haemolymph of the silkworm, Bombyx mori, by injection of formalin-treated Escherichia coli K-12 in the anterior and posterior body part of the ligated larvae.

1. Induction of antibacterial activity was investigated in the ligated fifth instar larvae of the silkworm, Bombyx mori, by injection of formalin-treated Escherichia coli K-12 into the haemocoel in the anterior and in the posterior body part, followed by activity determination by inhibition zone assay of the haemolymph at 12 and 24 hr after immunization. 2. At 12 hr after immunization, high antibacterial activity, approximately 6.8-7.5 mm in the anterior body part and 4.5-6.4 mm in the posterior body part in diameter of a clear zone (2.0 mm for no activity) was detectable in day 3 larvae. This result was in good contrast to expression of lectin gene in the ligated flesh fly, Sarcophaga peregrena (Shiraishi and Natori, 1988, FEBS Lett. 232, 163-166), in which only the anterior part of insect responded to stimulus of injury. 3. Antibacterial activity at 24 hr after immunization in days 3 or 4 ligated larvae was lower than that at 12 hr; 4.0-4.5 mm of activity was observed in the anterior body part, and no activity was observed in day 3 ligated larvae in the posterior body part. 4. Acid polyacrylamide gel electrophoresis of the haemolymph of immunized insects followed by overlay assay showed that the size of antibacterial activity bands were similar between the haemolymph from 12 hr and from 24 hr, and between the anterior and the posterior body part. This result was contradictory to the observation of activity by inhibition zone assay. The activity bands were associated with peptides that were similar to cecropin-like peptides A and B in the silkworm.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Oral immunization with Escherichia coli K-12 of the fifth instar larvae of the silkworm, Bombyx mori, reared on an artificial diet under completely aseptic conditions.

1. Effect of oral administration of live or formalin-treated Escherichia coli (E. coli) K-12 to the fifth instar, days 1 and 3 larvae of the silkworm, Bombyx mori, on induction of antibacterial activity in the haemolymph was investigated using the silkworms reared on an artificial diet under completely aseptic conditions. 2. When live E. coli was administered to the male day 1 larvae, low but significant antibacterial activity of 3.8 mm was detectable in the haemolymph of one individual at 48 hr after immunization. The proportion of the larvae to express antibacterial activity increased thereafter and at 120 hr after immunization, all three individuals showed antibacterial activity. In day 3 male larvae, activity was detectable at 48 and 72 hr after immunization. 3. When formalin-treated E. coli was orally administered to days 1 and 3 male larvae, no activity was detectable at any time post-immunization. 4. In the second experiment, when day 1 larvae, females and males were orally immunized with live E. coli, only females showed antibacterial activity in the haemolymph, beginning from 24 hr after immunization and up to 96 hr. 5. Removal of an antibiotic, chloramphenicol, from ingredients of an artificial diet was required for induction of antibacterial activity with oral administration of live E. coli. 6. When live E. coli that grows at pH 9.0 was selected and used for oral immunization, antibacterial activity was induced both in females and males at 72 hr after immunization and the activity was observed at 96 hr. 7. These results suggest that establishment of oral immunization with live E. coli in the silkworm larvae requires multiplication of E. coli in the midgut lumen and possibly its colonization on the luminal surface.

Administration, Oral↗

Genetic linkage groups in the Japanese brown frog (Rana japonica).

To elucidate the genetic linkage groups of the Japanese brown frog (Rana japonica) and compare them with those of other amphibians, we analyzed the inheritance of alleles at 15 enzyme and blood protein loci and one pigment locus (Blk) in 3,298 offspring derived from 37 crosses using 28 males heterozygous at these loci. Of 63 pairs of loci tested for linkage, 55 pairs showed independent assortment in all crosses examined. In another six pairs, that is, between FUM/ME-1, alpha-GDH/MDH-2, MDH-1/ME-1, MDH-2/ME-2, MDH-2/PEP-C, and ME-2/PEP-C, all the offspring analyzed were parental, and none were recombinant. In the other two pairs, that is, between GPI/PEP-D and AAT-1/Blk, most of the offspring were parental, and some were recombinants with recombination rates ranging from 4.8% to 8.0%. Thus the following four linkage groups comprising 11 loci were established in R. japonica. The first group included the loci for alpha-GDH, MDH-2, ME-2, and PEP-C; the second group included the loci for MDH-1, ME-1, and FUM; the third group included the loci for GPI and PEP-D; and the fourth group included the loci for AAT-1 and Blk. No linkage between the other five loci--ADA, MPI, PEP-A, PGM, and Alb--was observed in the present study.

Animals↗

TNF-alpha and endotoxin serum levels in cancer patients undergoing intraperitoneal hyperthermic perfusion.

Intraperitoneal hyperthermic perfusion (IPHP) is performed as one treatment for patients with advanced gastrointestinal cancer complicated by peritoneal dissemination or carcinomatous peritonitis. However, the anticancer mechanism of IPHP and its safety have not yet been fully elucidated. It has been experimentally known that endotoxinemia occurs by high body temperature, and that endotoxin stimulates the macrophage, monocyte and endothelial cell to induce the production of TNF-alpha. TNF-alpha is one of cytokines to be induced at the initial phase as a host immune response and play an important role to initiate the systemic inflammatory response syndrome (SIRS). We have tested whether the serum concentrations of TNF-alpha and endotoxin are elevated following IPHP. Eleven patients with gastro-intestinal cancer underwent surgery combined with IPHP. Mixed venous blood obtained from pulmonary artery (PA-blood) was collected at four sampling points. TNF-alpha and endotoxin levels in the PA-blood were measured by ELISA and a limulus amoebocyte lysate assay respectively. In all patients the serum TNF-alpha levels in PA-blood were temporarily elevated following IPHP from less than 10 pg/ml before IPHP to 42.8 +/- 26.6 pg/ml; endotoxin levels were not altered. This study shows that IPHP has the ability to induce endogenous TNF-alpha not mediated by endotoxin.

Adult↗

Integrated approach to the diagnosis of pancreatic carcinoma.

A prospective comparison of sensitivity, specificity and predictive value of ultrasonography (US), computed tomography (CT), endoscopic retrograde cholangiopancreatography (ERCP) and angiography was performed in 112 patients with proven pancreatic carcinoma. ERCP had the highest sensitivity and specificity in evaluation of pancreatic carcinoma. CT was the least sensitive in detecting pancreatic malignancy. Angiography had the highest positive predictive value. US and ERCP had a high negative predictive value. The initial examination of a patient with suspected pancreatic carcinoma is by US. Should this produce an abnormal result, CT is then used to confirm a lesion. A diagnosis of unresectable pancreatic carcinoma could be made by US and CT without the need for additional imaging procedures. If not, ERCP is indicated when there is some distinct reason to suspect pancreatic carcinoma. If ERCP reveals an abnormality, then angiography is performed to determine whether the lesion is benign or malignant, and if malignant whether it is resectable or not.

Adult↗

Expression of the Newcastle disease virus (NDV) fusion glycoprotein and vaccination against NDV challenge with a recombinant baculovirus.

The hybrid baculovirus constructed from Autographa california nuclear polyhedrosis virus (NPV) and Bombyx mori (silkworm) NPV was used for expression of fusion glycoprotein (F) of Newcastle disease virus (NDV) strain D26. The gene encoding F protein was introduced into the improved baculovirus expression vector derived from the host-range-expanded baculovirus. In Spodoptera frugiperda (fall armyworm) cells infected with a recombinant baculovirus, HyF121, the expressed F protein was properly located onto the cell surface. After silkworm pupae were infected with HyF121, a subunit vaccine against NDV was prepared from the HyF121-infected pupae. Chickens inoculated with the subunit vaccine were protected against virulent NDV challenge.

Animals↗