Enhanced Identification of Transcriptional Enhancers Provides Mechanistic Insights into Diseases.

Enhancers are distal cis-regulatory DNA elements that increase the expression of target genes. Various experimental and computational approaches including chromatin signature profiling have been developed to predict enhancers on a genome-wide scale, although each method has its advantages and disadvantages. Here we overview an emerging method to identify transcribed enhancers at exceedingly high nucleotide resolution based on enhancer RNA transcripts captured by Cap Analysis of Gene Expression (CAGE) technology. We further argue that disease-causative regulatory mutations at enhancers are increasingly recognized, emphasizing the importance of enhancer identification in functional and clinical genomics including, but not limited to, genome-wide association studies (GWASs) and cancer genomics studies.

Cytokine levels in sputum of patients with tracheostomy and profound multiple disabilities.

BACKGROUND: Airway immunopathogenesis is unclear in patients with profound multiple disabilities (PMD) who undergo tracheostomy. METHODS: The levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-6, IL-8, IL-10, and IL-12p70 cytokines were determined in sputum of 28 patients with PMD who underwent tracheostomy and in 14 healthy subjects, using a cytometric bead array. RESULTS: The concentrations of IL-1beta, IL-6 and IL-8 in the patients were significantly higher than those in controls (p < 0.001). IL-6, and IL-8 levels in eight PMD patients in the febrile period were significantly higher than those in the afebrile period (p < 0.01 and p < 0.05, respectively). Serum CRP levels were slightly elevated in 12 of the 28 patients (43%) in the afebrile period, but there were no significant differences in the level of any cytokine between patients with normal and elevated serum CRP. CONCLUSION: PMD patients with tracheostomy have chronic airway inflammation.

[Analysis of 11 patients with profound multiple disabilities who underwent laryngotracheal separation].

We examined the clinical usefulness of laryngotracheal separation surgery for patients with profound multiple disabilities (PMD). The subjects were 11 severely retarded children who experienced repeated incidents of aspiration pneumonia or who were enough to have aspiration pneumonia easily. A retrospective investigation of their medical records was performed regarding pre- and post-operative data, including the number of times sputum suctionings were required, the number of times pneumonia developed, respiratory conditions, and nutrition methods. The guardians were interviewed regarding musle tone, spasm, sleep quality, internal medications, and changes in mood. After surgery, improvement was confirmed in the number of times sputum suctionings were required, the incidence of pneumonia and respiratory conditions, and oral intake of food in three children. The guardians were aware of improvements in their children's sleep quality and mood. Laryngotracheal separation surgery can reduce the burden of health care for patients with PMD by improving their respiratory conditions and methods of nutrition intake.

[Low-dose therapy with carbamazepine for convulsions associated with mild gastroenteritis].

We investigated the effect of carbamazepine on convulsions associated with mild gastroenteritis. Sixteen infants and young children (aged 9 months to 3 years) who experienced repetitive convulsions associated with mild gastroenteritis were admitted to our hospital. We treated the sixteen affected patients with 5 mg/kg of carbamazepine once per day until the diarrhea had stopped. Thirteen of the sixteen patients were subjected to intravenous and/or suppository administration of diazepam (0.3-0.5 mg/kg/time), and one patient suppository administration of 0.5 mg/kg diazepam and 5.7 mg/kg phenobarbital before the administration of carbamazepine. In all patients who were given diazepam and/or phenobarbital, the convulsions recurred after the administration of these medicines. The convulsions occurred 2 to 8 times (mean, 4.1 times) before the administration of carbamazepine. Fifteen of the sixteen patients had no seizures after the administration of carbamazepine. One patient had one convulsion 15 min after the administration of carbamazepine. All patients were treated with 5 mg/kg of carbamazepine once per day until the diarrhea had stopped, i.e. for 2 to 9 days (mean, 6.4 days). Low dose therapy with carbamazepine once per day is thus effective for convulsions associated with mild gastroenteritis.

High-resolution solid-state MAS 73Ge NMR spectra of some organogermanes.

High-resolution solid-state magic angle spinning (73)Ge NMR spectra of some organogermanium compounds were measured. Most tetrasubstituted germanes with identical substituents exhibited signals except for one case. Tetrasubstituted germanes with two kinds of different but somewhat similar substituents exhibited broad peaks. Trisubstituted germanes failed to show signals, indicating the importance of symmetry around germanium.

[No association between moyamoya disease and polymorphism of IGF2R].

Moyamoya disease is a cerebrovascular disorder of unknown etiology. Its high incidence in East Asia and accumulation in family members suggest a genetic background. A high incidence of maternal inheritance implicates genomic imprinting in this disorder. Based on this hypothesis, we studied the association between moyamoya disease and IGF2R gene on chromosome 6, but found no evidence for such association between them. On the other hand, heterogeneous expressions of IGF2R were confirmed in the lymphocytes. Some individuals showed monoallelic expression and others showed biallelic expression.

[Molecular screening for moyamoya disease by use of expressed sequence tag on chromosome 3p].

Moyamoya disease is a well-known cerebrovascular disorder of unknown pathogenesis affecting terminal portion of internal carotid arteries and causing ischemic attacks. Its familial occurrence suggests genetic background. We hypothesized that paternally imprinted gene might be associated with this disorder. To identify the expressed sequence tags (ESTs) with monoallelic expressions on chromosome 3, we used mouse A9 hybrid cells having human chromosome 3. Two ESTs showed only maternal expression in mouse A9 hybrid cells, and four showed non-expression in the lymphocytes derived from moyamoya patients. Although these ESTs are clustered on the same 150 kb region, we finally failed to identify cDNA in this region.

High-resolution solid-state 73Ge NMR spectra of hexacoordinated germanium compounds.

High-resolution solid-state magic angle spinning 73Ge NMR spectra of hexavalent germanium compounds, i.e. diiodogermylene (1a) and bis(1-pyrrolyl)(meso-tetraphenylporphyrinato)germanium (3a), were determined. To the best of our knowledge, this is the first example of high-resolution solid-state 73Ge NMR spectra of hexacoordinated germanium nuclei. The symmetry requirement for observation of high-resolution solid-state 73Ge NMR spectra is discussed.

Safety from thromboembolism using intravenous immunoglobulin therapy in Kawasaki disease: Study of whole-blood viscosity.

BACKGROUND: High-dose intravenous infusions of immunoglobulin (IVIG) are well established as a standard therapy for Kawasaki disease (KD) for reducing the risk of coronary artery aneurysms. IVIG therapy might increase the blood viscosity both in vitro and in vivo, which has been reported as a risk factor for cardiovascular or cerebrovascular thromboembolism in adults. METHODS: We measured the whole-blood viscosity in vitro, serum IgG and albumin, and blood hematocrit in 10 patients with KD and 10 with non-KD (five with acute encephalitis, one with sepsis, one with idiopathic thrombocytopenic purpura, one with Guillain-Barré syndrome, one with insulin-dependent diabetes mellitus, and one with Evans syndrome) before and after IVIG therapy. RESULTS: The blood viscosity increased significantly after IVIG therapy in the patients with non-KD, but did not increase in those with KD. CONCLUSION: The results of the present study suggest that the use of IVIG therapy for KD might be relatively safe, with no risk of thromboembolism due to hyperviscosity.

NF-kappaB activation in cerebrospinal fluid cells from patients with meningitis.

We examined whether or not NF-kappaB, a factor that regulates expression of the genes that code for pro-inflammatory cytokines, is activated in cerebrospinal fluid (CSF) cells to investigate the production of pro-inflammatory cytokines by CSF cells in patients with meningitis. Western blotting demonstrated that NF-kappaB was more activated in CSF cells of patients with bacterial meningitis than in those of patients with aseptic meningitis. NF-kappaB was hardly activated in carcinomatous meningitis. The NF-kappaB activation in CSF cells of patients with meningitis tended to be correlated with the CSF interleukin-6 concentration. Our data suggested that CSF cells produce pro-inflammatory cytokines through NF-kappaB activation in meningitis, and that increased NF-kappaB activation in CSF cells indicate infectious meningitis rather than carcinomatous meningitis.