Development of formulae for dental age estimation using digital radiographs in the Mongolian population.

This study aimed to examine the applicability of the pulp/tooth ratio (PTR) method for age estimation in Mongolian populations using panoramic radiographs and derive new regression formulae. Moreover, we aimed to assess the accuracy of these formulae in other subjects from the Mongolian population and compare them with the other formulae derived from different Asian populations. The total sample size of the study was 381. The formulae were derived from the examination of panoramic radiographs of 271 individuals aged 15-62 years. Following Cameriere's method, PTR was calculated for the upper and lower canine teeth. Linear regression analyses were performed between the actual age and that obtained from upper-lower canine PTR and established formulae for age estimation. To verify the formulae, two types of test samples were collected: 73 panoramic radiographs and 37 periapical radiographs. The estimated age was calculated using our new formulae and three other formulae derived from Asian populations. The correlation coefficient between the actual age and that obtained by PTR was significantly negative for both canines. According to our new regression formulae, the differences between the estimated age and actual age showed a bell-shaped curve distribution in both test groups. While using the other formulae derived from the Asian population, the distribution patterns obtained were significantly different in the Mongolian population. This study was the first to examine the relationship between actual age and PTR in Mongolian population, and these results advance the field of forensic science in Mongolia.

A chemogenetic platform for controlling plasma membrane signaling and synthetic signal oscillation.

Chemogenetic methods enabling the rapid translocation of specific proteins to the plasma membrane (PM) in a single protein-single ligand manner are useful tools in cell biology. We recently developed a technique, in which proteins fused to an Escherichia coli dihydrofolate reductase (eDHFR) variant carrying N-terminal hexalysine residues are recruited from the cytoplasm to the PM using the synthetic myristoyl-d-Cys-tethered trimethoprim (mDcTMP) ligand. However, this system achieved PM-specific translocation only when the eDHFR tag was fused to the N terminus of proteins, thereby limiting its application. In this report, we engineered a universal PM-targeting tag for mDcTMP-induced protein translocation by grafting the hexalysine motif into an intra-loop region of eDHFR. We demonstrate the broad applicability of the new loop-engineered eDHFR tag and mDcTMP pair for conditional PM recruitment and activation of various tag-fused signaling proteins with different fusion configurations and for reversibly and repeatedly controlling protein localization to generate synthetic signal oscillations.

Analysis on unidentified cases in which dental information was collected from 2014 to 2019 in Miyagi Prefecture, Japan.

This study aimed to summarize the characteristics of unidentified cases in which dental information was collected during 2014-2019 in Miyagi Prefecture and to discuss the challenges and social characteristics of dental identification in routine forensic work. Cases were selected and examined from the database of dental charts collected at Tohoku University and the database of the Miyagi Prefectural Police. The annual percentage of cases with matched dental findings ranged from 19.2 - 37.1%, and 80.6% of the cases were identified within four days using dental information. The most common age group was the 50-70 s. Males were more common in the 20-70 s, whereas females were more frequent in the 80-90 s age group. Utilization of dental information was lower for cases involving young people and those over 90 years of age. The number of unidentified cases and unspecified death cases in the central block was significantly higher than other regions. However, there were no significant differences in sex, age, and season between each region. Spearman's rank-order analysis of the unspecified deaths showed a strong correlation with the monthly average temperature (ρ = 0.89, p < 0.01). Within the limitations of this study, the results provide valuable information on the long-term progress of routine dental identification work and could be useful as a reference for a statistical survey. Future research must examine the role of dental information in forensic identification by exploring the role of more specific background characteristics among autopsy cases.

Chemogenetic Control of Protein Localization and Mammalian Cell Signaling by SLIPT.

Chemical control of protein localization is a powerful approach for manipulating mammalian cellular processes. Self-localizing ligand-induced protein translocation (SLIPT) is an emerging platform that enables control of protein localization in living mammalian cells using synthetic self-localizing ligands (SLs). We recently established a chemogenetic SLIPT system, in which any protein of interest fused to an engineered variant of Escherichia coli dihydrofolate reductase, DHFRiK6, can be rapidly and specifically translocated from the cytoplasm to the inner leaflet of the plasma membrane (PM) using a trimethoprim (TMP)-based PM-targeting SL, mDcTMP. The mDcTMP-mediated PM recruitment of DHFRiK6-fusion proteins can be efficiently returned to the cytoplasm by subsequent addition of free TMP, enabling temporal and reversible control over the protein localization. Here we describe the use of this mDcTMP/DHFRiK6-based SLIPT system for inducing (1) reversible protein translocation and (2) synthetic activation of the Raf/ERK pathway. This system provides a simple and versatile tool in mammalian synthetic biology for temporally manipulating various signaling molecules and pathways at the PM.

An Improved Intracellular Synthetic Lipidation-Induced Plasma Membrane Anchoring System for SNAP-Tag Fusion Proteins.

The ability to chemically introduce lipid modifications to specific intracellular protein targets would enable the conditional control of protein localization and activity in living cells. We recently developed a chemical-genetic approach in which an engineered SNAP-tag fusion protein can be rapidly relocated and anchored from the cytoplasm to the plasma membrane (PM) upon post-translational covalent lipopeptide conjugation in cells. However, the first-generation system achieved only low to moderate protein anchoring (recruiting) efficiencies and lacked wide applicability. Herein, we describe the rational design of an improved system for intracellular synthetic lipidation-induced PM anchoring of SNAP-tag fusion proteins. In the new system, the SNAPf protein engineered to contain an N-terminal hexalysine (K6) sequence and a C-terminal 10-amino acid deletion, termed K6-SNAPΔ, is fused to a protein of interest. In addition, a SNAP-tag substrate containing a metabolic-resistant myristoyl-DCys lipopeptidomimetic, called mDcBCP, is used as a cell-permeable chemical probe for intracellular SNAP-tag lipidation. The use of this combination allows significantly improved conditional PM anchoring of SNAP-tag fusion proteins. This second-generation system was applied to activate various signaling proteins, including Tiam1, cRaf, PI3K, and Sos, upon synthetic lipidation-induced PM anchoring/recruitment, offering a new and useful research tool in chemical biology and synthetic biology.