Full-Length Sequence of Mouse Acupuncture-Induced 1-L (Aig1l) Gene Including Its Transcriptional Start Site

We have been investigating the molecular efficacy of electroacupuncture (EA), which is one type of acupuncture therapy. In our previous molecular biological study of acupuncture, we found an EA-induced gene, named acupuncture-induced 1-L (Aig1l), in mouse skeletal muscle. The aims of this study consisted of identification of the full-length cDNA sequence of Aig1l including the transcriptional start site, determination of the tissue distribution of Aig1l and analysis of the effect of EA on Aig1l gene expression. We determined the complete cDNA sequence including the transcriptional start site via cDNA cloning with the cap site hunting method. We then analyzed the tissue distribution of Aig1l by means of northern blot analysis and real-time quantitative polymerase chain reaction. We used the semiquantitative reverse transcriptase-polymerase chain reaction to examine the effect of EA on Aig1l gene expression. Our results showed that the complete cDNA sequence of Aig1l was 6073 bp long, and the putative protein consisted of 962 amino acids. All seven tissues that we analyzed expressed the Aig1l gene. In skeletal muscle, EA induced expression of the Aig1l gene, with high expression observed after 3 hours of EA. Our findings thus suggest that the Aig1l gene may play a key role in the molecular mechanisms of EA efficacy

Herbal Medicine Containing Licorice May Be Contraindicated for a Patient with an HSD11B2 Mutation

Licorice ingestion, as well as mutations in the HSD11B2 gene, inhibits 11β-hydroxysteroid dehydrogenase type 2 (11βHSD2) enzyme activity, causing the syndrome of apparent mineral corticoid excess (AME). However, the combined effect of licorice ingestion and an HSD11B2 mutation has never been reported, until now. In this study, we demonstrated that licorice ingestion can produce overt hypertension in an individual without medical history of hypertension who is heterozygous for wild-type and mutant HSD11B2 genes. Our patient was a 51-year-old female with serious hypertension who had been taking herbal medicine containing licorice for more than one year. She was clinically diagnosed as having licorice intoxication, because she did not present with hypertension after ceasing the herbal medicine. Molecular analysis showed that she carried a missense mutation, c.40C>T, in HSD11B2. In conclusion, licorice ingestion is an environmental risk factor for hypertension or AME state in patients with a mutation in HSD11B2. Carrying a mutation in HSD11B2 is, conversely, a genetic risk factor for licorice-induced hypertension or AME state. Herbal medicine containing licorice may, therefore, be contraindicated in patients with an HSD11B2 mutation

A Homozygous Mutation in UGT1A1 Exon 5 May Be Responsible for Persistent Hyperbilirubinemia in a Japanese Girl with Gilbert’s Syndrome

The UGT1A1 gene encodes a responsible enzyme, UDP-glucuronosyltransferase1A1, for bilirubin metabolism. Many mutations have already been identified in patients with inherited disorders with hyperbilirubinemia, Crigler-Najjar syndrome and Gilbert’s syndrome. In this study, we identified a UGT1A1 mutation in an 8-year-old Japanese girl with persistent hyperbilirubinemia who was clinically diagnosed as having Gilbert’s syndrome. For the mutational analysis of UGT1A1, we performed a full sequence analysis of the gene using the patient’s DNA. She was homozygous for a T to G transversion at nucleotide position 1456 in UGT1A1 exon 5 (c.1456T>G), leading to the substitution of aspartate for tyrosine at position 486 of the UGT1A1 protein (p.Y486D). In conclusion, the homozygous mutation of UGT1A1 may be responsible for persistent hyperbilirubinemia in this patient

The C677 Mutation in Methylene Tetrahydrofolate Reductase Gene: Correlation with Uric Acid and Cardiovascular Risk Factors in Elderly Korean men

The C677T mutation in the methylene tetrahydrofolate reductase (MTHFR) gene results in elevated homocysteine levels and, presumably, in increased cardiovascular risk. Moreover, elevated homocysteine levels are reportedly associated with high serum uric acid levels. We evaluated the MTHFR genotype and a panel of biochemical, hematological variables, and lifestyle characteristics in 327 elderly Korean men (age range 40-81 yr; mean, 51.87). This study shows that mutation of the MTHFR gene may be a risk for hyperuricemia. The mean uric acid levels for the C/C, C/T and T/T genotypes were 5.54, 5.91 and 6.33 mg/dL, respectively (p=0.000). The T/T genotype was significantly more frequent in subjects with high uric acid levels (p=0.003). Thus, this mutation of the MTHFR gene is implied by the study results to be a risk factor of hyperuricemia in elderly Korean men. However, the relationship between the MTHFR mutation and uric acid metabolism remains unclear. Therefore, further studies are necessary to explain the associated between the MTHFR mutation and elevated uric acid levels, and to examine potential relationships between it and conventional cardiovascular risk factors

Comparison of the Bronchodilative Effects of Salbutamol Delivered via Three Mesh Nebulizers in Children with Bronchial Asthma

Background: We compared the bronchodilative effects of salbutamol delivered via 3 different mesh nebulizers, Aeroneb-go®(AE), Omron-NE-U22®(OM) and Pari-eMotion®(PA). Methods: We enrolled 36 children with asthma who visited the Kurosaka Pediatrics and Allergy Clinic, randomly assigned to 3 groups for treatment with AE, OM or PA. The dose of salbutamol in the solution was 0.15 mg x body weight (kg) (minimum 2.5 mg, maximum 5 mg). FEVi, PEFR and V50 were measured in these patients before treatment, and at 15 and 30 minutes after salbutamol inhalation using one of the 3 mesh nebulizers. Results: All groups showed a significant improvement of FEVi, PEFR and V50 at 30 minutes after salbutamol inhalation. The AE group did not show a significant improvement in PEFR at 15 minutes after inhalation, whereas a significant improvement in FEVi and V50 was evident at the same time point. The OM group showed no significant improvement in V50 at 15 minutes after inhalation, whereas this group clearly showed a significant improvement in PEFR and FEVi at the same time point. Conclusions: Overall, all 3 mesh nebulizers were useful devices in treating bronchial asthma, although some differences in lung function improvement were evident. The limitation of this study is that subjects did not include patients with severe asthma attacks

Study Abroad Experience Is Related to Japanese Doctors' Behavior to See Foreign Patients

Globalization in Japan involves increases in the number of foreign residents. While there are some English-speaking Japanese doctors that are willing to see foreign patients, many are reluctant to do so. In this study, we attempted to clarify the factors that encourage Japanese doctors to see foreign patients. We conducted a questionnaire survey among medical doctors in Kobe City, Japan. The questionnaire was distributed to 172 doctors, and we received 139 responses. Statistical analysis showed a significant correlation between the frequency of seeing foreign patients and having the experience of studying abroad (p<0.05), confirming our hypothesis. There was also a significant correlation between having the experience of studying abroad and the doctors’ self-evaluations of their English ability (p<0.05). There was no significant correlation found, however, between the frequency of seeing foreign patients and that of reading English research articles. These data suggested that the experience of living abroad rather than the exposure to English research articles was more highly correlated with seeing greater numbers of foreign patients. In conclusion, greater exposure to colloquial English was one of the determinants of the doctors’ greater willingness to see foreign patients. In the Japanese medical education curriculum, therefore, it would be necessary to offer alternatives to studying abroad for those students who do not have such opportunities

An Antisense Oligonucleotide against a Splicing Enhancer Sequence within Exon 1 of the MSTN Gene Inhibits Pre-mRNA Maturation to Act as a Novel Myostatin Inhibitor

Antisense oligonucleotides (ASOs) are agents that modulate gene function. ASO-mediated out-of-frame exon skipping has been employed to suppress gene function. Myostatin, encoded by the MSTN gene, is a potent negative regulator of skeletal muscle growth. ASOs that induce skipping of out-of-frame exon 2 of the MSTN gene have been studied for their use in increasing muscle mass. However, no ASOs are currently available for clinical use. We hypothesized that ASOs against the splicing enhancer sequence within exon 1 of the MSTN gene would inhibit maturation of pre-mRNA, thereby suppressing gene function. To explore this hypothesis, ASOs against sequences of exon 1 of the MSTN gene were screened for their ability to reduce mature MSTN mRNA levels. One screened ASO, named KMM001, decreased MSTN mRNA levels in a dose-dependent manner and reciprocally increased MSTN pre-mRNA levels. Accordingly, KMM001 decreased myostatin protein levels. KMM001 inhibited SMAD-mediated myostatin signaling in rhabdomyosarcoma cells. Remarkably, it did not decrease GDF11 mRNA levels, indicating myostatin-specific inhibition. As expected, KMM001 enhanced the proliferation of human myoblasts. We conclude that KMM001 is a novel myostatin inhibitor that inhibits pre-mRNA maturation. KMM001 has great promise for clinical applications and should be examined for its ability to treat various muscle-wasting conditions