Oral delivery of novel human IGF-1 bioencapsulated in lettuce cells promotes musculoskeletal cell proliferation, differentiation and diabetic fracture healing.

Human insulin-like growth factor-1 (IGF-1) plays important roles in development and regeneration of skeletal muscles and bones but requires daily injections or surgical implantation. Current clinical IGF-1 lacks e-peptide and is glycosylated, reducing functional efficacy. In this study, codon-optimized Pro-IGF-1 with e-peptide (fused to GM1 receptor binding protein CTB or cell penetrating peptide PTD) was expressed in lettuce chloroplasts to facilitate oral delivery. Pro-IGF-1 was expressed at high levels in the absence of the antibiotic resistance gene in lettuce chloroplasts and was maintained in subsequent generations. In lyophilized plant cells, Pro-IGF-1 maintained folding, assembly, stability and functionality up to 31 months, when stored at ambient temperature. CTB-Pro-IGF-1 stimulated proliferation of human oral keratinocytes, gingiva-derived mesenchymal stromal cells and mouse osteoblasts in a dose-dependent manner and promoted osteoblast differentiation through upregulation of ALP, OSX and RUNX2 genes. Mice orally gavaged with the lyophilized plant cells significantly increased IGF-1 levels in sera, skeletal muscles and was stable for several hours. When bioencapsulated CTB-Pro-IGF-1 was gavaged to femoral fractured diabetic mice, bone regeneration was significantly promoted with increase in bone volume, density and area. This novel delivery system should increase affordability and patient compliance, especially for treatment of musculoskeletal diseases.

Design and verification of shielding for the advanced spent fuel conditioning process facility.

An Advanced spent fuel Conditioning Process Facility (ACPF) has recently been constructed by a modification of previously unused cells. ACPF is a hot cell with two rooms located in the basement of the Irradiated Materials Experiment Facility (IMEF) at the Korea Atomic Energy Research Institute. This is for demonstrating the advanced spent fuel conditioning process being proposed in Korea, which is an electrolytic reduction process of spent oxide fuels into a metallic form. The ACPF was designed with a more than 90 cm thick high density concrete shield wall to handle 1.38 PBq (37,430 Ci) of radioactive materials with dose rates lower than 10 muSv h in the operational areas (7,000 zone) and 150 muSv h in the service areas (8,000 zone). In Monte Carlo calculations with a design basis source inventory, the results for the bounding wall showed a maximum of 3 muSv h dose rate at an exterior surface of the ACPF for gamma radiation and 0.76 muSv h for neutrons. All the bounding structures of the ACPF were investigated to check on the shielding performance of the facility to ensure the radiation safety of the facility. A test was performed with a 2.96 TBq (80 Ci) 60Co source unit and the test results were compared with the calculation results. A few failure points were discovered and carefully fixed to meet the design criteria. After fixing the problems, the failure points were rechecked and the safety of the shielding structures was confirmed. In conclusion, it was confirmed that all the investigated parts of the ACPF passed the shielding safety limits by using this program and the ACPF is ready to fulfill its tasks for the advanced spent fuel conditioning process.

Characterization of chromosomal breakpoints in an ALL patient using cross-species color banding.

Cross-species color-banded karyotype (Rx-FISH) results were compared with those of conventional G-banded metaphases from the same sample. Breakpoints and karyotype were confirmed as 46,XX,t(8;22)(q24;q11), der(9)t(1;9)(q21;p13) through the novel technology of cross-species color banding in an acute leukemic patient (ALL, L3); the karyotype was 46,XX,t(8;22)(q24;q11),der(9)t(1;9)(q25;p24) by conventional G-banding.

Genetic alterations of gastric cancer: comparative genomic hybridization and fluorescence In situ hybridization studies.

Genetic changes leading to the development of gastric cancers are still in dispute. In the following study, we used comparative genomic hybridization (CGH) to screen for DNA copy number changes along all chromosomes in 37 gastric carcinomas, and fluorescence in situ hybridization (FISH) with the C-MYC and TP53 probes in 14 cases for comparison. The aim of this study was to identify those chromosome regions that contain genes important for the development of gastric carcinomas and to identify genetic markers associated with tumor progression. The most often involved gains were 2q, 7pq, 8pq, 13q, 17q, 18q, and 20pq. The most commonly deleted regions were 17p. The pattern of genetic changes was different depending on the existence of nodal metastasis and histologic types. Gains in 8q and losses in 17p were the most common features of the CGH changes. However, only 3 among the available 10 cases (30%) showed an amplification of the C-MYC gene by FISH. Allelic loss of TP53 was found in 2 of 4 cases (50%). This difference might be due to another rearrangement of these 2 genes which cannot be detected by FISH, or other possible genes in that area may be involved in the tumorigenesis and nodal metastasis of gastric carcinomas.

Genetic alterations in hepatocellular carcinoma and intrahepatic cholangiocarcinoma.

In the following study, we used comparative genomic hybridization (CGH) to screen and compare for genetic alterations of hepatocellular carcinoma (HCC) and intrahepatic choalgiocarcinoma (ICC). The studies showed distinctive features of genetic alterations between the two tumors. Characteristic abnormal changes for HCC were 1q gain and loss of 4q, 10q and 13q regions. In contrast, gains of 5p, 7p, 13q and 20q were more predominant in ICC. Losses of 16q, 17p, and 18q, and gain of 8q region showed a similar high frequency of incidence in both tumors. The most striking and different findings were 1q amplification in HCC and 20q gain in ICC. Our data indicate that ICC shows the pattern of genetic alterations similar to pancreatic and colorectal cancers. This suggests that the genetic alterations in tumorigenesis show a similar pattern depending on the origin of cells, not the organ.

Detection of genetic alterations in bladder tumors by comparative genomic hybridization and cytogenetic analysis.

Comparative genomic hybridization (CGH) and conventional cytogenetic karyotyping were used to screen for losses and gains of DNA sequences along all chromosome arms in 16 bladder tumors. Cytogenetic results were highly complex. The most frequently affected chromosomes were 5, 8, 9, 21, and Y as determined by karyotyping. There was close correlation between the CGH data and cytogenetic results in near-diploid tumors with simple karyotypes. However, some unexpected results were observed by CGH in tumors with several composite clones. Common amplification of copy numbers of DNA sequences by CGH were seen at 1q, 3q, 4q, 5p, 6p/q, 7p, 8q, 11q, 12q, 13q, 17q, 18q, and 20p/q (more than 20% of cases). High level amplification was noted at 1p32, 3p21, 3q24, 4q26, 8q21-qter, 11q14-22, 12q15-21, 12q21-24, 13q21-31, 17q22, and 18q22. Deletions were noted at 2q21-qter. 4q13-23, 5q, 8p12-22, 9p/q, and 11p13-15 (more than 20% of cases). Although most amplifications and deletions have been previously described in the literature, our study showed some intriguing and uncommon regions, different from those found in past studies. These were the amplification of 7p, 8q, 11q14-qter 12q24-24, 13q21-31, and 18q22, and deletion on 4q13-23, even though loss of heterozygosity was not detected at this locus. In spite of the very complex pattern of genetic changes in bladder tumors, most of these uncommon aberrations have to be implicated in bladder tumors, and further molecular genetic methods are necessary to establish whether the chromosomal regions contain candidate genes which contributed to the initiation and progression of bladder tumors.

Synthesis and antimicrobial activity of new 3 alpha-hydroxy-23,24-bisnorcholane polyamine carbamates.

3 alpha-Hydroxy-23,24-bisnorcholane spermidine and spermine carbamates 2-7 have been synthesized and their antimicrobial and hemolytic activities were evaluated. They exhibited excellent in vitro activities especially against methicillin-resistant Staphylococcus aureus.

Synthesis and antimicrobial activity of squalamine analogue.

Synthesis and antimicrobial activity of squalamine analogue 2 are reported. The synthesis of 2 was accomplished from bisnoralcohol 3. The spermidine moiety was introduced via reductive amination of an appropriately functionalized 3beta-aminosterol with spermidinyl aldehyde 17 utilizing sodium triacetoxyborohydride as the reducing agent. Compound 2 shows weaker antimicrobial activity than squalamine.