Protective effect of astaxanthin nanoemulsion on mammalian inner ear hair cells.

Background: Aminoglycoside antibiotics are used for treating certain acute infections. However, these drugs cause ototoxicity by inducing inner ear hair cell death. Aims/Objectives: We investigated the protective effect of a nanoemulsion of the carotenoid astaxanthin on mammalian inner ear hair cells against neomycin-induced ototoxicity. Material and Methods: Dose-response relationship, quantification of hair cell loss, and reactive oxygen species production were assayed in response to neomycin with and without astaxanthin in cultured utricles of CBA/N mice. In addition, auditory brain response (ABR) and hair cell loss after exposure to the nanoformulation and loud noise were examined in vivo in guinea pigs. Results: Astaxanthin suppressed neomycin-induced reduction of hair cells by reducing the production of hydroxy radicals. Furthermore, hair cell loss in the second rotation of the cochlea was significantly lower in the astaxanthin group than in the noise-only group. Conclusions and Significance: The blood-labyrinth barrier limits the successful delivery of drugs for inner ear complications. However, in the nanoemulsion form, astaxanthin can penetrate the round window (fenestra ovale) membrane, enabling topical administration. Thus, astaxanthin nanoemulsion could be useful in treating ototoxicity in individuals with inner ear complications.

Prevention of progressive hearing loss in a mouse model of diabetes by oral intake of eicosapentaenoic acid ethyl ester.

BACKGROUND: The prevalence of hearing impairment in patients with diabetes was significantly higher, and the development of preventive methods is desirable. AIMS/OBJECTIVES: This study examined the effects of eicosapentaenoic acid (EPA) administration on the prevention of early hearing loss in diabetic mice. MATERIALS AND METHODS: Tsumura, Suzuki, Obese Diabetes (TSOD) mice were used as a model of diabetes and Tsumura, Suzuki, Non Obesity (TSNO) mice were used as controls. The animals were divided into three groups: the TSNO group and the TSOD (EPA-) group (provided sunflower oil), and the TSOD (EPA+) group (provided EPA). Auditory brainstem responses (ABRs) were measured and the cochlea was evaluated histologically. RESULTS: The TSOD (EPA+) group showed a lower tendency to increase thresholds than the TSOD (EPA-) group. The TSOD (EPA+) group had a significantly lower ABR threshold than the TSOD (EPA-) group from 11 to 14 months of age at 4 kHz. Narrowing of the capillary lumens in the stria vascularis and thickening of the vessel wall in the modiolus were observed in the TSOD (EPA-) group. CONCLUSIONS AND SIGNIFICANCE: It was suggested that the suppression of cochlear vascular atherosclerosis by EPA administration in TSOD mice suppressed early age-related hearing loss.

Novel compound heterozygous mutations in the PLEC gene in a neonate with epidermolysis bullosa simplex with pyloric atresia.

Epidermolysis bullosa (EB) is a heterogeneous group of inherited disorders characterized by the blistering of the skin and mucous membranes. Although the molecular basis of EB has been significantly elucidated, the precise phenotypes of the lethal types of EB have not been completely characterized. Herein, we report a severe case of EB with pyloric atresia (PA). The patient was a Japanese boy who not only had skin lesions but also various complications such as PA, dysphagia, hypotonia, infectious keratitis with corneal ulcer, obstructive uropathy and protein-losing enteropathy. Genetic analysis led to the identification of two novel compound heterozygous mutations in the last exon of the plectin (PLEC) gene. Based on this finding, EB simplex with PA was diagnosed. Immunostaining with anti-plectin antibodies revealed truncated plectin proteins lacking the C-terminus in the patient's skin. We also conducted a prenatal diagnosis in subsequent pregnancy. Our report further highlights the crucial role of plectin in many organs and provides valuable information regarding the phenotypes resulting from mutations in the PLEC gene.

Effect of preservation on the physical and chemical properties of the temporal fascia.

OBJECTIVE: The temporal fascia has been widely used in tympanoplasty. In addition, the preserved fascia has been also used in the ear surgery. In this study, we planned the experiments to determine whether physical and chemical properties of the fascia preserved at a low temperature. METHODS: Preserved temporal fasciae from 21 patients were used in this study. The thickness of the temporal fascia was measured under a 3D laser microscope. The tensile strength was evaluated using a tensile tester. In addition, the chemical property evaluated was the biologic antioxidative potential of samples. RESULTS: The results showed that the strength of the fascia was not affected by the retention period. The thick fascia tended to show the less tensile strength. The intensity was highest in middle-aged donors when compared to young and older donor. The antioxidative potentials did not affect the preservation. CONCLUSION: The results suggested that the preserved temporal fascia could be safely used for tympanoplasty.

Oral administration of an herbal medicine to prevent progressive hearing loss in a mouse model of diabetes.

OBJECTIVE: Tsumura Suzuki Obese Diabetes (TSOD) mice exhibit early age-associated hearing loss. Histopathological analysis of these mice shows narrowing of capillaries in the stria vascularis and chronic reduction of blood flow in the cochlea. In this study, we investigated the effect of oral administration of a herbal medicine or calorie restriction on hearing in TSOD mice. METHODS: TSOD mice were divided into 4 groups: CR (calorie restriction), BF and DS (treated with the herbal medicines, Bofutsushosan and Daisaikoto, respectively), and the control group. Body weight, blood glucose levels, and auditory brainstem responses (ABRs) were measured. The cochleae were excised and evaluated histopathologically. RESULTS: Blood glucose levels were suppressed in the CR, BF, and DS groups. In addition, the elevation of ABR thresholds was inhibited in the CR, BF, and DS groups. Cochlear blood vessels remained wide in the three treatment groups compared with the control group. These results suggested that the administration of these herbal medicines improved glucose tolerance and yielded results similar to those on calorie restriction. CONCLUSION: Oral administration of 2 herbal medicines can prevent hearing function disorder in a model mouse of diabetes. The results may clarify the possibility of clinical application.

A study of hearing function and histopathologic changes in the cochlea of the type 2 diabetes model Tsumura Suzuki obese diabetes mouse.

OBJECTIVES: This study used Tsumura Suzuki Obese Diabetes (TSOD) mice as a spontaneous type 2 diabetes model and Tsumura Suzuki Non-obesity (TSNO) mice as controls to investigate factors involved in the onset of hearing impairment. METHOD: Body weight, blood glucose levels, and auditory brainstem responses (ABRs) were measured. The cochleae were excised and evaluated histopathologically. RESULTS: The TSOD mice showed significant hyperglycemia at 2-7 months and severe obesity at 5-10 months; significantly elevated ABR thresholds at 8-10 months; and the capillary lumens in the cochlea stria vascularis were narrower in the TSOD mice than in the TSNO mice. At 17 months, India ink vascular staining of the TSOD mice's cochleae revealed decreased capillary density in the stria vascularis. The vascular area of capillaries in the stria vascularis and the vascular area were significantly smaller in TSOD mice. Histopathological analysis showed vessel wall thickening in the modiolus and narrowed capillaries in the stria vascularis, suggesting reduced blood flow to the inner ear. CONCLUSION: The diabetes mice model used in our study showed early age-associated hearing loss, and histopathology showed findings of vessel wall thickening in the modiolus, narrowing of capillaries in the stria vascularis, and chronically reduced blood flow in the cochlea.

[Intravascular large B-cell lymphoma with low signal intensity lesions on T2 weighted spinal magnetic resonance image that were suspected to be hemorrhages].

A 79-year-old female had a spinal lesion that was definitely diagnosed as intravascular large B-cell lymphoma on the basis of skin biopsy findings, and she was treated by rituximab-containing chemotherapy. The spinal lesion showed high and low signal intensities on T2 weighted magnetic resonance imaging (MRI) scans, those low signal intensity lesions were suspected to be hemorrhages. The hemorrhages were thought to have been caused by interaction between atypical lymphoma cells and the endothelial cells of spinal blood vessels, by hemorrhagic infarction or by rupture of the capillary endothelium due to interaction between rituximab and lymphoma cells. Intravascular large B-cell lymphoma cases rarely show low signal intensity on spinal T2 weighted MRI scans.

Involvement of CD56brightCD11c+ cells in IL-18-mediated expansion of human γδ T cells.

γδ T cells are considered to be innate lymphocytes that play an important role in host defense against tumors and infections. We recently reported that IL-18 markedly amplified γδ T cell responses to zoledronate (ZOL)/IL-2. In an extension of this finding, we analyzed the mechanism underlying the IL-18-mediated expansion of γδ T cells. After incubation of PBMCs with ZOL/IL-2/IL-18, the majority of the cells expressed γδ TCR, and the rest mostly exhibited CD56(bright)CD11c(+) under the conditions used in this study. CD56(bright)CD11c(+) cells were derived from a culture of CD56(int)CD11c(+) cells and CD14(+) cells in the presence of IL-2 and IL-18 without the addition of ZOL. They expressed IL-18Rs, HLA-DR, CD25, CD80, CD83, CD86, and CD11a/CD18. In addition, they produced IFN-γ, TNF-α, but not IL-12, when treated with IL-2/IL-18, and they exerted cytotoxicity against K562 cells, thus exhibiting characteristics of both NK cells and dendritic cells. Incubation of purified γδ T cells with CD56(bright)CD11c(+) cells in the presence of ZOL/IL-2/IL-18 resulted in the formation of massive cell clusters and led to the marked expansion of γδ T cells. However, both conventional CD56(-/int)CD11c(high) dendritic cells induced by GM-CSF/IL-4 and CD56(+)CD11c(-) NK cells failed to support the expansion of γδ T cells. These results strongly suggest that CD56(bright)CD11c(+) cells play a key role in the IL-18-mediated proliferation of γδ T cells.

Crystallization and X-ray diffraction studies of DNA-free and DNA-bound forms of EcoO109I DNA methyltransferase.

EcoO109I DNA methyltransferase (M.EcoO109I) is a type II modification enzyme from the EcoO109I restriction-modification system identified in Escherichia coli strain H709c. M.EcoO109I recognizes double-stranded RGGNCCY (where R = A or G, Y = T or C and N is any base) and transfers a methyl group to the C5 of the inner cytosines from S-adenosylmethionine. To reveal the mechanism of substrate recognition by M.EcoO109I, DNA-free and DNA-bound forms of M.EcoO109I were successfully crystallized. Crystals of the DNA-free and DNA-bound forms belonged to space groups P4(2)2(1)2, with unit-cell parameters a = b = 120.5, c = 79.8 Å, and P2(1), with unit-cell parameters a = 55.8, b = 77.4, c = 117.4 Å, ß = 93.5°, respectively.

Crystallization and preliminary X-ray crystallographic analyses of EcoO109I and its complex with DNA.

EcoO109I is a type II restriction endonuclease that recognizes seven base pairs of the degenerate and discontinuous sequence RGGNCCY. The enzyme and its complex with DNA were successfully crystallized by the hanging-drop vapour-diffusion method using polyethylene glycols as precipitants. The crystal of EcoO109I belongs to space group I4, with unit-cell parameters a = b = 175.5, c = 44.6 angstoms, and that of the DNA complex belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 49.1, b = 71.8, c = 203.2 angstroms. Full sets of X-ray diffraction data from the enzyme and its complex with DNA were collected to 2.4 and 1.9 angstroms resolution, respectively.

[Surveillance on concurrent administration of quinolones and anti-inflammatory drugs in a community hospital].

Since convulsions associated with the concurrent administration of enoxacin and fenbufen were reported in 1986, the concurrent administration of quinolones and anti-inflammatory drugs has been regulated to be contraindicated or carefully administered. However, the real incidence of the co-administration is not clear. We surveyed the incidence of co-administration of these drugs from the prescriptions in a community hospital. Quinolones were prescribed in 1% of the out-patients, and anti-inflammatory drugs were co-administrated in 16% of quinolone-prescribed patients. Among quinolones prescribed, levofloxacin was used most frequently. And acetaminphen (including acetaminophen-combination) was most frequently prescribed with quinolones, and anti-inflammatory drugs, which is regulated to be carefully administrated with quinolones, were frequently co-prescribed. In medical practice, quinolones were revealed to be co-administered with anti-inflammatory drugs. Since our recent report suggests that each quinolone and each anti-inflammatory drug has different activity in their drug interaction, it should be necessary to recognize the interaction of these drugs separately for their effective and safe use in clinical field.

C.EcoO109I, a regulatory protein for production of EcoO109I restriction endonuclease, specifically binds to and bends DNA upstream of its translational start site.

The EcoO109I restriction-modification system, which recognizes 5'-(A/G)GGNCC(C/T)-3', has been cloned, and contains convergently transcribed endonuclease and methylase. The role and action mechanism of the gene product, C.EcoO109I, of a small open reading frame located upstream of ecoO109IR were investigated in vivo and in vitro. The results of deletion analysis suggested that C.EcoO109I acts as a positive regulator of ecoO109IR expression but has little effect on ecoO109IM expression. Assaying of promoter activity showed that the expression of ecoO109IC was regulated by its own gene product, C.EcoO109I. C.EcoO109I was overproduced as a His-tag fusion protein in recombinant Escherichia coli HB101 and purified to homogeneity. C.EcoO109I exists as a homodimer, and recognizes and binds to the DNA sequence 5'-CTAAG(N)(5)CTTAG-3' upstream of the ecoO109IC translational start site. It was also shown that C.EcoO109I bent the target DNA by 54 +/- 4 degrees.