Agonist-independent GPCR activity regulates anterior-posterior targeting of olfactory sensory neurons.

G-protein-coupled receptors (GPCRs) are known to possess two different conformations, active and inactive, and they spontaneously alternate between the two in the absence of ligands. Here, we analyzed the agonist-independent GPCR activity for its possible role in receptor-instructed axonal projection. We generated transgenic mice expressing activity mutants of the ß2-adrenergic receptor, a well-characterized GPCR with the highest homology to odorant receptors (ORs). We found that mutants with altered agonist-independent activity changed the transcription levels of axon-targeting molecules--e.g., Neuropilin-1 and Plexin-A1--but not of glomerular segregation molecules--e.g., Kirrel2 and Kirrel3--thus causing shifts in glomerular locations along the anterior-posterior (A-P) axis. Knockout and in vitro experiments demonstrated that Gs, but not Golf, is responsible for mediating the agonist-independent GPCR activity. We conclude that the equilibrium of conformational transitions set by each OR is the major determinant of expression levels of A-P-targeting molecules.

Comparative clinical outcomes and radiological images of clavicle hook plate versus Scorpion Plate® for unstable distal clavicle fractures.

BACKGROUND: Although various surgical methods are available for unstable distal clavicle fractures, consensus remains lacking on the optimal technique. Therefore, the present retrospective study aimed to compare the clavicle hook plate and Scorpion Plate® in terms of clinical outcomes and radiological findings for unstable distal clavicle fractures. METHODS: Fifty-seven patients with unstable distal clavicle fractures who underwent treatment using a clavicle hook plate (Group H; 28 patients) or Scorpion Plate® (Group S; 29 patients) were included in the present study. No patients received additional augmentation and all were followed-up for >1 year (mean follow-up, 28 months). Clinical outcomes were operation time, bleeding volume, complications, range of motion (ROM) at 6 months after surgery and final follow-up, and clinical scores (Constant-Murley score and University of California, Los Angeles (UCLA) shoulder score). Radiological outcomes assessed were coracoclavicular distance (CCD), difference in CCD between affected and non-affected sides (ΔCCD), and acromioclavicular subluxation ratio (%ACS) from plain X-rays. The χ2 test and Mann-Whitney U test were used to compare each outcome. RESULTS: Complications were seen in 3 shoulders (10.7%) in Group H. ROM was significantly worse in Group H at 6 months postoperatively, but no significant differences between groups were evident at final follow-up. Moreover, no significant differences in clinical outcomes were seen between groups. In terms of radiological results, Group H showed greater improvement in CCD and ΔCCD than Group S. Further, %ACS was significantly worse in Group S. CONCLUSIONS: The clavicle hook plate allows reconstruction of a more anatomical position than the Scorpion Plate®, but carries a greater risk of complications. Conversely, the Scorpion Plate® has a low risk of complications, but acromioclavicular subluxation remains. However, no significant differences in ROM or clinical outcomes were apparent at final follow-up.

Glucokinase Variant Proteins Are Resistant to Fasting-Induced Uridine Diphosphate Glucose-Dependent Degradation in Maturity-Onset Diabetes of the Young Type 2 Patients.

We previously reported that glucokinase undergoes ubiquitination and subsequent degradation, a process mediated by cereblon, particularly in the presence of uridine diphosphate glucose (UDP-glucose). In this context, we hereby present evidence showcasing the resilience of variant glucokinase proteins of maturity-onset diabetes of the young type 2 (MODY2) against degradation and, concomitantly, their influence on insulin secretion, both in cell lines and in the afflicted MODY2 patient. Hence, glucose-1-phodphate promotes UDP-glucose production by UDP-glucose pyrophosphorylase 2; consequently, UDP-glucose-dependent glucokinase degradation may occur during fasting. Next, we analyzed glucokinase variant proteins from MODY2 or persistent hyperinsulinemic hypoglycemia in infancy (PHHI). Among the eleven MODY2 glucokinase-mutated proteins tested, those with a lower glucose-binding affinity exhibited resistance to UDP-glucose-dependent degradation. Conversely, the glucokinaseA456V-mutated protein from PHHI had a higher glucose affinity and was sensitive to UDP-glucose-dependent degradation. Furthermore, in vitro studies involving UDP-glucose-dependent glucokinase variant proteins and insulin secretion during fasting in Japanese MODY2 patients revealed a strong correlation and a higher coefficient of determination. This suggests that UDP-glucose-dependent glucokinase degradation plays a significant role in the pathogenesis of glucose-homeostasis-related hereditary diseases, such as MODY2 and PHHI.

Differentiation Capacity of Porcine Skeletal Muscle-Derived Stem Cells as Intermediate Species between Mice and Humans.

Large animal experiments are important for preclinical studies of regenerative stem cell transplantation therapy. Therefore, we investigated the differentiation capacity of pig skeletal muscle-derived stem cells (Sk-MSCs) as an intermediate model between mice and humans for nerve muscle regenerative therapy. Enzymatically extracted cells were obtained from green-fluorescence transgenic micro-mini pigs (GFP-Tg MMP) and sorted as CD34+/45- (Sk-34) and CD34-/45-/29+ (Sk-DN) fractions. The ability to differentiate into skeletal muscle, peripheral nerve, and vascular cell lineages was examined via in vitro cell culture and in vivo cell transplantation into the damaged tibialis anterior muscle and sciatic nerves of nude mice and rats. Protein and mRNA levels were analyzed using RT-PCR, immunohistochemistry, and immunoelectron microscopy. The myogenic potential, which was tested by Pax7 and MyoD expression and the formation of muscle fibers, was higher in Sk-DN cells than in Sk-34 cells but remained weak in the latter. In contrast, the capacity to differentiate into peripheral nerve and vascular cell lineages was significantly stronger in Sk-34 cells. In particular, Sk-DN cells did not engraft to the damaged nerve, whereas Sk-34 cells showed active engraftment and differentiation into perineurial/endoneurial cells, endothelial cells, and vascular smooth muscle cells, similar to the human case, as previously reported. Therefore, we concluded that Sk-34 and Sk-DN cells in pigs are closer to those in humans than to those in mice.

R1526 residue in arginine/proinsulin binding domain of UGGT1 is involved in proinsulin binding.

Arginine releases proinsulin from binding to UGGT1 in the endoplasmic reticulum (ER). PSIPRED analysis showed that the arginine/proinsulin binding domain (A/PBD) in the C-terminal region of UGGT1 forms a disordered region, which is flexible and outside of the main protein structure. Both arginine and proinsulin may easily access the disordered region of A/PBD. Using the SNP library, two variants, Q1518∗ and R1526C, were identified in this region. UGGT1Q1518∗ protein is a deficient form of A/PBD and ER-retention signal (ERRS). UGGT1Q1518∗ protein in cell analysis reveals that mutated protein is mainly secreted from the cells because it lacks ERRS. We found another UGGT1 variant, UGGT1R1526C. At the molecular level, less interaction of proinsulin with UGGT1 was observed in both human UGGT1R1526C and mouse UGGT1L1518C with/without arginine. However, UGGT1R1526C and UGGT1WT interact with arginine similarly. We identified several amino acid residues for the arginine and proinsulin interaction. Here, the R1526 residue of UGGT1 is involved in proinsulin-interaction and is not involved in arginine-interaction.

Paraneoplastic cerebellar degeneration with anti-Yo antibodies and an associated submandibular gland tumor: a case report.

BACKGROUND: As a debilitating syndrome, paraneoplastic cerebellar degeneration (PCD) remains challenging to treat. Further, anti-Yo antibody (directed against human cerebellar degeneration-related protein 2) detection in patients with PCD is associated with unsatisfactory responses to existing therapies. Here, we present the case of a 60-year-old woman who developed PCD with anti-Yo antibodies and a submandibular gland tumor. CASE PRESENTATION: A 60-year-old woman presented with a 5-day history of unsteadiness of gait and inadequate coordination of her extremities, along with truncal instability. Although walking without aid was possible, dysmetria of all four limbs, trunk, and gait ataxia was observed. While routine biochemical and hematological examinations were normal, the patient's blood was positive for anti-Yo antibodies. When the neurological symptoms deteriorated despite administration of intravenous methylprednisolone, fluorodeoxyglucose-positron emission tomography (FDG-PET) and computed tomography (CT) images with contrast enhancement were performed, which showed a tumor in the left submaxillary gland. She underwent total left submandibular gland resection, including the tumor; histological and immunohistochemical results revealed a salivary duct carcinoma. She was administered intravenous methylprednisolone, followed by 10 plasma exchange sessions, intravenous immunoglobulins, and cyclophosphamide therapy. Following treatment, her symptoms were not alleviated, even after the reduction of anti-Yo titers. CONCLUSIONS: Although tumor detection was delayed, early tumor detection, diagnosis, and PCD treatment are essential because any delay can result in the progression of the disorder and irreversible neurological damage. Therefore, we recommend that the possibility of a salivary gland tumor should be considered, and whole-body dual-modality CT, including the head and neck, and FDG-PET should be performed at the earliest for patients with well-characterized paraneoplastic antibodies when conventional imaging fails to identify a tumor.

Long-term outcomes of open modified inferior capsular shift for traumatic anterior shoulder instability: over 20 years of follow-up.

BACKGROUND: This study aimed to assess the long-term outcomes of the open modified inferior capsular shift procedure across more than 20 years in patients with traumatic anterior shoulder instability. METHODS: Participants in this study comprised 84 patients (86 shoulders; 67 men, 17 women). Mean follow-up was 28.0 years. We compared the recurrent instability rate after surgery, apprehension, revision rate, satisfaction, return to preinjury sporting activity, and patient background characteristics between recurrent and nonrecurrent groups. Twenty-seven patients who were examined directly were evaluated for differences in range of motion and muscle strength between affected and nonaffected sides. Clinical outcome scores used for this study were the Western Ontario Shoulder Instability Index and the Rowe score. RESULTS: Nine shoulders (10.5%) showed recurrent instability, as dislocation in 3 shoulders and subluxation in 6 shoulders, but no revisions were required. All patients with recurrent instability were males with new trauma. Three shoulders developed recurrent instability ≤5 years after surgery, and the remaining 6 shoulders showed recurrent instability >5 years after surgery. No significant differences in any patient characteristic were identified between the recurrent and nonrecurrent groups. Significant differences between affected and nonaffected sides were seen in the mean active range of motion or muscle strength for external rotation in the anatomic position or in 90° of abduction. Western Ontario Shoulder Instability Index and Rowe score were significantly worse in the recurrent group than in the nonrecurrent group. Patients reported that 84 shoulders (97.7%) were "much better." Most athletes (88.4%) had returned to sports activity at a level >70%. CONCLUSION: We investigated long-term outcomes of the open modified inferior capsular shift procedure for traumatic anterior shoulder instability. Our data suggest that recurrent instability might result from new trauma even if a long time has passed since the open modified inferior capsular shift procedure, so follow-up should be continued as long as possible after surgery. As in other reports, satisfaction was high and clinical scores were good. We thus believe this surgical method offers good results even after more than 20 years.

Medical Image Sharing in Japan.

This paper reports the history, background including politics, current status of Japan's health imaging study and other information sharing. Its realization was slow until the Ministry of Health, Labour and Welfare (MHLW) started paying digital image storage at the same rate as films in 2008. Information sharing was initiated in early 2010s, which was before vendors became ready for Integrating the Healthcare Enterprise (IHE) cross-enterprise document sharing (XDS), with the result that most of 34 large regional sharing systems are in non-standardized protocol. One standardized example is the Hamamatsu area where inexpensive online PDI (portable data for imaging) was introduced.

UDP-Glucose: A Cereblon-Dependent Glucokinase Protein Degrader.

We previously reported that glucokinase is ubiquitinated and degraded by cereblon with an unknown endogenous glucokinase protein degrader. Here, we show that UDP-glucose is a glucokinase protein degrader. We identified that both glucose and UDP-glucose bind to glucokinase and that both uridine and UDP-glucose bind to cereblon in a similar way to thalidomide. From these results, UDP-glucose was identified as a molecular glue between cereblon and glucokinase. Glucokinase produces glucose-6-phosphate in the pancreas and liver. Especially in ß-cells, glucokinase is the main target of glucose for glucose-induced insulin secretion. UDP-glucose administration ubiquitinated and degraded glucokinase, lowered glucose-6-phosphate production, and then reduced insulin secretion in ß-cell lines and mice. Maturity-onset diabetes of the young type 2 (MODY2) glucokinaseE256K mutant protein was resistant to UDP-glucose induced ubiquitination and degradation. Taken together, glucokinase ubiquitination and degradation signaling might be impaired in MODY2 patients.

Chorionic gonadotropin stimulates maternal hepatocyte proliferation during pregnancy.

The liver increases its size during pregnancy to adapt to metabolic demand associated with pregnancy. Our previous study showed that proliferation of maternal hepatocytes are increased during pregnancy in mice and that estradiol (E2) is one of the candidate hormones responsible for maternal hepatocyte proliferation. Here, we discovered that chorionic gonadotropin (CG) induces maternal hepatocyte proliferation during pregnancy. CG administration was sufficient to stimulate hepatocyte proliferation in non-pregnant mice as well as in cell culture system. We conclude that CG stimulates proliferation in the early pregnancy of maternal hepatocytes. In contrast, estrogen stimulates hepatocyte proliferation in the late pregnancy.

EHR2CCAS: A framework for mapping EHR to disease knowledge presenting causal chain of disorders - chronic kidney disease example.

OBJECTIVE: The goal of this work was to capture diseases in patients by comprehending the fine-grained medical conditions and disease progression manifested by transitions in medical conditions. We realize this by introducing our earlier work on a state-of-the-art knowledge presentation, which defines a disease as a causal chain of abnormal states (CCAS). Here, we propose a framework, EHR2CCAS, for constructing a system to map electronic health record (EHR) data to CCAS. MATERIALS AND METHODS: EHR2CCAS is a framework consisting of modules that access heterogeneous EHR to estimate the presence of abnormal states in a CCAS for a patient in a given time window. EHR2CCAS applies expert-driven (rule-based) and data-driven (machine learning) methods to identify abnormal states from structured and unstructured EHR data. It features data-driven approaches for unlocking clinical texts and imputations based on the EHR temporal properties and the causal CCAS structure. This study presents the CCAS of chronic kidney disease as an example. A mapping system between the EHR from the University of Tokyo Hospital and CCAS of chronic kidney disease was constructed and evaluated against expert annotation. RESULTS: The system achieved high prediction performance in identifying abnormal states that had strong agreement among annotators. Our handling of narrative varieties in texts and our imputation of the presence of an abnormal state markedly improved the prediction performance. EHR2CCAS presents patient data describing the temporal presence of abnormal states in CCAS, which is useful in individual disease progression management. Further analysis of the differentiation of transition among abnormal states outputted by EHR2CCAS can contribute to detecting disease subtypes. CONCLUSION: This work represents the first step toward combining disease knowledge and EHR to extract abnormality related to a disease defined as fine-grained abnormal states and transitions among them. This can aid in disease progression management and deep phenotyping.

Locking versus nonlocking superior plate fixations for displaced midshaft clavicle fractures: A prospective randomized trial comparing clinical and radiografic results.

BACKGROUND: It is unknown whether locking or nonlocking superior plate fixation is better for managing displaced midshaft clavicle fractures. Therefore, we aimed to compare the clinical and radiographic outcomes of locking and nonlocking superior plate fixation of displaced midshaft clavicle fractures. METHODS: A total of 102 consecutive patients with displaced midshaft clavicle fractures (2B1 and 2B2 in Robinson classification) participated in this randomized controlled trial; 12 patients were excluded. Surgeries were performed using a 3.5-mm Locking Compression Plate (LCP) between 2007 and 2015. Patients were treated either with a locking plate (group L, n = 45) or a nonlocking plate (group N, n = 45). In both groups, the plates were fixed to the proximal and distal clavicle with two and/or three screws, respectively. The main outcome measures were complication rates, time to bone union, and Constant score. RESULTS: Forty-two patients in group L (mean age, 45.9 years) and 41 in group N (mean age, 43.6 years) were followed. The overall complication rates in groups L and N were 7.2% (three peri-implant fractures) and 7.3% (non-union, deformed plate, and peri-implant fracture), respectively (p = .98). The average time to union significantly differed between groups (L vs. N: 13.0 ± 4.1 vs. 17.5 ± 6.3 weeks; p < .01). However, the Constant score at the final follow-up was not significantly different between groups (L vs. N: 87.0 ± 12.3 vs. 89.8 ± 9.1). CONCLUSIONS: Similar complication rates and clinical results were found for locking and nonlocking superior plate fixation for displaced midshaft clavicle fractures. However, the time to bone union was shorter with the locking plate. This study suggests that both plating systems are effective for treating displaced midshaft clavicle fractures. LEVEL OF EVIDENCE: Therapeutic, level I.

Automatic Classification of Electronic Nursing Narrative Records Based on Japanese Standard Terminology for Nursing.

In Japan, nursing records are not easily put to secondary use because nursing documentation is not standardized. In recent years, electronic health records have necessitated the creation of Japanese nursing terminology. The purpose of this study was to develop and evaluate an automatic classification system for narrative nursing records using natural language processing technology and machine learning. We collected a week's worth of narrative nursing records from an academic hospital. The authors independently annotated the text data, dividing it into morphemes, the smallest meaningful unit in a language. During preprocessing when creating feature quantities, we used a Japanese tokenizer, MeCab, an open-source morphological parser, and the bag-of-words model. A support vector machine was adopted as a classifier for machine learning. The accuracy was 0.96 and 0.86 on the training set and test set, respectively, and the F value was 0.82. Our findings provide useful information regarding the development of an automatic classification system for Japanese nursing records using nursing terminology and natural language processing techniques.

UGGT1 retains proinsulin in the endoplasmic reticulum in an arginine dependent manner.

We sought to clarify a pathway by which L- and dD-arginine simulate insulin secretion in mice and cell lines and obtained the following novel two findings. (1) Using affinity magnetic nanobeads technology, we identified that proinsulin is retained in the endoplasmic reticulum (ER) through UDP-glucose:glycoprotein glucosyltransferase 1 (UGGT1) when arginine availability is limited. (2) L- and d-arginine release proinsulin from UGGT1 through competition with proinsulin and promote exit of proinsulin from the ER to Golgi apparatus. The ability of arginine to release proinsulin from UGGT1 closely correlates with arginine-induced insulin secretion in several models of ß cells indicating that UGGT1-proinsulin interaction regulates arginine-induced insulin secretion.

Highly accurate and explainable detection of specimen mix-up using a machine learning model.

Background Delta check is widely used for detecting specimen mix-ups. Owing to the inadequate specificity and sparseness of the absolute incidence of mix-ups, the positive predictive value (PPV) of delta check is considerably low as it is labor consuming to identify true mix-up errors among a large number of false alerts. To overcome this problem, we developed a new accurate detection model through machine learning. Methods Inspired by delta check, we decided to conduct comparisons with the past examinations and broaden the time range. Fifteen common items were selected from complete blood cell counts and biochemical tests. We considered examinations in which ≥11 among the 15 items were measured simultaneously in our hospital; we created individual partial time-series data of the consecutive examinations with a sliding window size of 4. The last examinations of the partial time-series data were shuffled to generate artificial mix-up cases. After splitting the dataset into development and validation sets, we allowed a gradient-boosting-decision-tree (GBDT) model to learn using the development set to detect whether the last examination results of the partial time-series data were artificial mixed-up results. The model's performance was evaluated on the validation set. Results The area under the receiver operating characteristic curve (ROC AUC) of our model was 0.9983 (bootstrap confidence interval [bsCI]: 0.9983-0.9985). Conclusions The GBDT model was more effective in detecting specimen mix-up. The improved accuracy will enable more facilities to perform more efficient and centralized mix-up detection, leading to improved patient safety.

Arginine induces IGF-1 secretion from the endoplasmic reticulum.

Arginine is a semi-essential amino acid with multiple functions, including stimulating the secretion of growth hormone (GH) and insulin-like growth factor 1 (IGF-1). IGF-1, which is produced by hepatocytes, plays important roles in cellular metabolism, proliferation, and growth. Previous studies showed that arginine-induced IGF-1 secretion occurs via two pathways: a GH-dependent pathway and an arginine-dependent pathway with an unknown mechanism. In this study, we identified the mechanisms regulating IGF-1 secretion. First, GH stimulates the translation of IGF-1 and increases IGF-1 protein levels, leading to IGF-1 secretion. As observed in fasted mice and hepatocytes cultivated in arginine-depleted medium, decreases in arginine concentrations resulted in IGF-1 retention in the endoplasmic reticulum. Arginine administration reverses this retention, leading to IGF-1 secretion. These data describe a novel IGF-1 secretion control system in the endoplasmic reticulum.

Estrogen induces estrogen receptor α expression and hepatocyte proliferation in late pregnancy.

Partial hepatectomy (PH) induces estradiol production, and then hepatocyte proliferation. Estradiol may play a role in triggering hepatocyte proliferation after PH. In this study, estradiol was injected to the Estrogen Receptor alpha (ERα) or ERß KO mice. No increased hepatocyte proliferation was observed in ERα KO mice, indicates that ERα is involved in estradiol-induced hepatocyte proliferation. The ERα and ERß KO mice are sterile, hence it is impossible to study ERα and ERß function during pregnancy when the estrogen levels are highest. Using conditional mutagenesis technique, we made ERα hepatocyte KO mice, which are fertile. We used these mice for analyzing the hepatocyte ERα function during pregnancy. However, in the control mice, the maternal hepatocyte was proliferated higher in late pregnancy, but no pregnancy-induced hepatocyte proliferation was observed in KO mice. Hence, we conclude that the maternal hepatocyte ERα is involved in estradiol-induced hepatocyte proliferation in late pregnancy.

A Trunk Stabilization Exercise Warm-up May Reduce Ankle Injuries in Junior Soccer Players.

The aim of this study was to investigate the efficacy of a trunk stabilization exercise warm-up program in reducing the incidence of lower extremity injuries among male junior soccer players. Two junior soccer teams participated in this study. The intervention (INT) team performed three trunk stabilization exercises before practice sessions and games, while a control (CON) team performed their usual warm-up without trunk exercises. Both teams engaged in regular soccer training and games, and were followed for the incidence of injury. As a result, overall injury incidence rates (IRs) were 2.65 injuries/1,000 h and 4.94 injuries/1,000 h in the INT and CON teams, respectively (incidence rate ratio [IRR]=0.54, 95% confidence interval [CI]=0.32-0.89, p=0.013). The IR of acute injuries was significantly lower in the INT team (1.91 injuries/1,000 h) than in the CON team (4.06 injuries/1,000 h) (IRR=0.47, 95%CI=0.26-0.84, p=0.009). Regarding injury sites, the IRs of ankle injuries in the INT team (0.32 injuries/1,000 h) were significantly lower than that in the CON team (2.28 injuries/1,000 h) (IRR=0.14, 95%CI=0.04-0.47, p<0.001). These results suggest that a warm-up program comprising trunk stabilization exercises alone can prevent acute injuries, especially ankle injuries.

Selective fluorescence detection method for selenide and selenol using monochlorobimane.

The low redox potential of selenide and selenol is physiologically important, as it confers efficient catalytic abilities to selenoproteins. Quantitative determination of selenol and selenide provide important clues for understanding the metabolism and physiological function of selenium. However, selective detection of selenol and selenide is extremely difficult because of their chemical similarity to thiol and sulfide. In this study, we established a highly sensitive, selective, quantitative, and simple method for detection of selenol and selenide, using a reaction with monochlorobimane (MCB), followed by ethyl acetate extraction of the product syn-(methyl,methyl)bimane. We analyzed selenide production from selenite, catalyzed by human glutathione reductase, and also determined selenide and selenol concentrations in Hepa1-6 cells using the MCB method, to demonstrate its practical applications. This study provides a new tool for selenium detection in biology.

Construction of functional neuronal circuitry in the olfactory bulb.

Recent studies using molecular genetics, electrophysiology, in vivo imaging, and behavioral analyses have elucidated detailed connectivity and function of the mammalian olfactory circuits. The olfactory bulb is the first relay station of olfactory perception in the brain, but it is more than a simple relay: olfactory information is dynamically tuned by local olfactory bulb circuits and converted to spatiotemporal neural code for higher-order information processing. Because the olfactory bulb processes ∼1000 discrete input channels from different odorant receptors, it serves as a good model to study neuronal wiring specificity, from both functional and developmental aspects. This review summarizes our current understanding of the olfactory bulb circuitry from functional standpoint and discusses important future studies with particular focus on its development and plasticity.