Strong Evidence of a Combination Polymorphism of the Tyrosine Kinase 2 Gene and the Signal Transducer and Activator of Transcription 3 Gene as a DNA-Based Biomarker for Susceptibility to Crohn’s Disease in the Japanese Population

OBJECTIVE: An association between susceptibility to inflammatory bowel disease (IBD) and polymorphisms of both the tyrosine kinase 2 gene (TYK2) and the signal transducer and activator of transcription 3 gene (STAT3) was examined in a Japanese population in order to identify the genetic determinants of IBD. METHODS: The study subjects comprised 112 patients with ulcerative colitis, 83 patients with Crohn's disease (CD), and 200 healthy control subjects. Seven tag single-nucleotide polymorphisms (SNPs) in TYK2 and STAT3 were detected by PCR-restriction fragment length polymorphism. RESULTS: The frequencies of a C allele and its homozygous C/C genotype at rs2293152 SNP in STAT3 in CD patients were significantly higher than those in control subjects (P = 0.007 and P = 0.001, respectively). Furthermore, out of four haplotypes composed of the two tag SNPs (rs280519 and rs2304256) in TYK2, the frequencies of a Hap 1 haplotype and its homozygous Hap 1/Hap1 diplotype were significantly higher in CD patients in comparison to those in control subjects (P = 0.023 and P = 0.024, respectively). In addition, the presence of both the C/C genotype at rs2293152 SNP in STAT3 and the Hap 1/Hap 1 diplotype of TYK2 independently contributes to the pathogenesis of CD and significantly increases the odds ratio to 7.486 for CD (P = 0.0008). CONCLUSION: TYK2 and STAT3 are genetic determinants of CD in the Japanese population. This combination polymorphism may be useful as a new genetic biomarker for the identification of high-risk individuals susceptible to CD

Identification of a shootin1 isoform expressed in peripheral tissues

Shootin1 is a brain-specific cytoplasmic protein involved in neuronal polarity formation and axon outgrowth. It accumulates at the leading edge of axonal growth cones, where it mediates the mechanical coupling between F-actin retrograde flow and cell adhesions as a clutch molecule, thereby producing force for axon outgrowth. In this study, we report a novel splicing isoform of shootin1 which is expressed not only in the brain but also in peripheral tissues. We have renamed the brain-specific shootin1 as shootin1a and termed the novel isoform as shootin1b. Immunoblot and immunohistochemical analyses with a shootin1b-specific antibody revealed that shootin1b is distributed in various mouse tissues including the lung, liver, stomach, intestines, spleen, pancreas, kidney and skin. Interestingly, shootin1b immunoreactivity was widely detected in epithelial cells that constitute simple and stratified epithelia; in some cells, it colocalized with E-cadherin and cortactin at cell–cell contact sites. Shootin1b also localized in dendritic cells in the spleen. These results suggest that shootin1b may function in various peripheral tissues including epithelial cells

Double-layered antibiotic-loaded cement spacer as a novel alternative for managing periprosthetic joint infection: an in vitro study

Abstract Background Previous studies comparing antibiotic-loaded calcium phosphate cement to polymethylmethacrylate cement reported that although the former has higher elution volumes over a longer period, it is mechanically weak when used alone. To counter this problem, a double-layered antibiotic-loaded cement spacer in which calcium phosphate cement is coated with polymethylmethacrylate cement was created. Methods In this study, we compared the double-layered spacer to the polymethylmethacrylate cement spacer in terms of eluent antibiotic concentration, bioactivity against methicillin-resistant Staphylococcus aureus, and mechanical strength. Double-layered and polymethylmethacrylate cement spacers that were loaded with vancomycin (VCM) were prepared and immersed in phosphate buffer for 84 days. To facilitate VCM elution from calcium phosphate cores in double-layered spacers, we also drilled multiple holes into the calcium phosphate layer from the spacer surface. Results We found that VCM concentrations in double-layered spacer eluents were higher than those in polymethylmethacrylate cement spacer eluents. The double-layered spacer also had higher bioactivity than the polymethylmethacrylate cement spacer. Although the polymethylmethacrylate cement spacer eluent lost the ability to inhibit bacterial growth on day 56, the double-layered spacer eluent maintained this ability for the duration of our study. Finally, the double-layered spacer retained high mechanical strength throughout the study period. Conclusions The beneficial biomechanical and drug-eluting properties of the double-layered spacer might qualify it to serve as a promising biomaterial that could be used for managing periprosthetic joint infections

Synthesis of [211At]MABG using remote‐controlled synthesizer and quality evaluation

ObjectivesPheochromocytoma is a known neuroendocrine tumor. Around 10% of pheochromocytomas are malignant and metastasize to bone, lung, etc. It is a refractory disease lacking an effective treatment. For malignant pheochromocytoma, treatment using with meta‐[131I]iodobenzylguanidine ([131I]MIBG) has been performed, but its therapeutic effect is limited. By applying α‐rays with a higher biological effect and shorter range than β‐rays to targeted radioisotope therapy, not only higher therapeutic effect can be expected but also radiation damage to normal tissues can be minimized. Therefore, meta‐[211At]astatobenzylguanidine ([211At]MABG) has been developed using astatine‐211 (211At, half‐life: 7.2 h), which emits α‐rays and has chemical properties similar to iodine.1 Its high accumulation in tumors and strong tumor volume‐reducing effect in pheochromocytoma model mice have already been reported.2 Currently, Our facility is planning to develop [211At]MABG for human clinical research in collaboration with the National Institutes for Quantum and Radiological Science and Technology (QST), which reported the therapeutic effect in mice. The aim is to synthesize [211At]MABG with a remotely controlled synthesizer using in‐house produced 211At at the Fukushima Medical University (FMU). MethodsWe produced 211At using 209Bi (α, 2n) 211At reaction by the MP‐30 cyclotron in FMU. The purification was carried out by dry distillation and obtained 211At as chloroform solution. An [211At]MABG‐remotely controlled synthesizer, designed and produced by the QST group, was installed in the glove box. 211At solution (13.5–141.4 MBq, ca. 0.1 mL) was introduced into the synthesizer and [211At]MABG was synthesized from the precursor, meta‐trimethylsilylbenzylguanidine hemisulfate (0.25 mg, 0.92 μmol) by electrophilic substitution reaction with N‐chlorosuccinimide (NCS, 50 μL as saturated solution in methanol) and trifluoroacetic acid (TFA, 0.35 mL) at 70°C for 10 min. Purification was carried out by solid phase extraction (Sep‐Pak tC18 Plus Light Cartridge, Waters). After washing with water (0.5 mL), [211At]MABG was eluted with 5% ethanol aqueous solution (2 mL). The quality of [211At]MABG was confirmed using radio‐HPLC. ResultsWhen [211At]MABG was synthesized using the synthesizer, the radiochemical yield was 59 ± 5%, and the synthesis time was 42 ± 2 minutes (n = 3). The maximum yield was 61.4 MBq (EOS). By solid‐phase extraction, it was possible to remove most of regents including NCS, precursor, and unreacted 211At. The radiochemical purity was >95%. ConclusionsWe succeeded in synthesizing [211At]MABG using 211At produced in FMU, and stable production was enabled in the yield and the synthesis time by using the synthesizer. REFERENCESVaidyanathan G, Zalutsky MR. Bioconjug Chem 1992;3:499.Ohshima Y, Sudo H, Watanabe S, Nagatsu K, Tsuji AB, Sakashita T, et al. Eur J Nucl Med Mol Imaging 2018; 45: 999–1010ISRS201