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Aim Olfactory impairment as a prodromal symptom, as well as sarcopenia, frailty and dependence as geriatric syndromes, is often associated with cognitive decline in older adults with progression of Alzheimer's disease. The present study aimed to evaluate the associations of olfactory and cognitive decline with these geriatric syndromes, and with structural changes of the brain in older adults.

An examination of taste and olfactometry in Yakumo-Cho inhabitants’ examination carried out in 2005, and the result reported in Academia Journal of Medicinal Plants 2018. This study examined olfactory function. A personal function test is calculated from the Yakumo study database, and the odor stick identification tests administered to healthy older adults. The participants were community dwellers who voluntarily participated in the Yakumo Study and had managed everyday life by themselves. The participants were engaged in a variety of jobs, not only white-collar but also agriculture, fishery,and forestry. Therefore, the city regarded as a representative of today's Japanese society. From the database, 298 participants (169 females and 129 males) were selected form data in August, 2019. The Odor Stick Identification Test (OSIT-J) was used to assess odor perception. The aromas used in the OSIT-J include curry, perfume, Japanese cypress, India ink, menthol, rose, wood, Smelly socks/ sweat, fried garlic, condensed milk, gas for cooking, and Japanese mandarin aromas.

Gustatory function is closely related to chewing and swallowing; however, there are currently no reports regarding gustatory function in persons with cerebral palsy (CP). The aim of this study was to compare the gustatory function between persons with CP and healthy controls. We investigated sweet, salty, sour and bitter tastes using the whole-mouth method and measured the electrogustometric thresholds in the chorda tympani nerve area. Twelve participants with CP (6 women and 6 men) for whom gustatory testing was possible at our facilities, and 17 age-matched controls (9 women and 8 men) were included. The mean age ± standard deviation was 58.6 ± 8.1 years and 58.5 ± 8.7 years in subjects with CP and controls, respectively. Taste detection and identification were significantly worse in persons with CP compared with the controls. Taste identification was more impaired than taste detection. At the highest concentration, taste identification was impossible 11 times out of 48 (12 persons × 4 kinds of tastes) in persons with CP but such a deficit was not observed in 68 attempts (17 persons × 4 kinds of tastes) involving controls. The electrogustometric thresholds were not significantly different between the groups. Gustatory function associated with chewing and swallowing is worse in persons with CP compared to the controls.

Geobacillus kaustophilus is a thermophilic bacterium that grows at temperatures ranging between 42 and 74 °C. Here, we modified this organism to produce the thermolabile protein (PyrFA) or its thermostable variant (PyrFV) and analyzed the transcriptome and growth efficiency profiles of the resultant strains. In the producer of PyrFA, the transcriptome profile was changed to facilitate ATP synthesis from NADH without pooling reduced quinones. This change implies that PyrFA production at elevated temperatures places an energy burden on cells potentially to maintain protein homeostasis. This was consistent with the observation that the PyrFA producer grew slower than the PyrFV producer at > 45 °C and had a lower cellular fitness. Similar growth profiles were also observed in the PyrFA and PyrFV producers derived from another thermophile (Geobacillus thermodenitrificans) but not in those from Escherichia coli at 30 °C. Thus, we suggest that the production of thermolabile proteins impairs host survival at higher temperatures; therefore, thermophiles are under evolutionary selection for thermostable proteins regardless of whether their functions are associated with survival advantages. This hypothesis provides new insights into evolutionary protein selection in thermophiles and suggests an engineering approach to select thermostable protein variants generated via random gene mutagenesis.

Hearing loss is a risk factor for cognitive impairment, and the use of a hearing aid (HA) may prevent cognitive decline alongside hearing loss. We aimed to elucidate the prevalence of self-reported HA usage in Japanese community-dwelling elders with hearing loss, and the effect of hearing and HA on cognitive impairment.A total of 1193 participants, who had audiometric defined hearing loss and were aged 60 years or over, had their cumulative 3260 observations followed up for 10 years from a large cohort of a Japanese study. Association between hearing (pure-tone average threshold level at 500, 1000, 2000, and 4000 Hz from the better hearing ear: PTABHE) and HA usage with cognitive impairment (total score of Mini-Mental State Estimation was under 27 or diagnosed as dementia) was analyzed using generalized estimating equations.The HA usage rate of the 1193 community-dwelling elders with hearing loss was 6% during the first involvement. The majority (59.2%) of HA users always used an HA. HA usage rate was 0.7% for the mild hearing loss group and 32.4% for the moderate or greater hearing loss group in the latest participating wave. PTABHE was significantly associated with cognitive impairment (odds ratio for every 10 dB 1.36; 95% CI 1.21-1.53, p0.0001) after adjusting for age, sex, education, depressed mood, smoking status, alcohol intake, income, activity, obesity, histories of hypertension, dyslipidemia, ischemic heart disease, diabetes, stroke, ear disease, and occupational noise exposure. PTABHE was also significantly associated with cognitive impairment in the mild hearing loss group (odds ratio for every 10 dB 1.34; 95% CI 1.05-1.72, p = 0.020) and moderate hearing loss group (odds ratio for every 10 dB 1.82; 95% CI 1.27-2.61, p = 0.001). HA use showed a significant suppressive effect on cognitive impairment in those with moderate hearing loss who always use an HA (odds ratio 0.54; 95% CI 0.30-1.00, p = 0.049).The prevalence of HA usage among Japanese community-dwelling elders with hearing loss is consistent, at around 10%. The hearing level remained a primary risk factor for cognitive impairment among elders with hearing loss after adjusting for several confounding factors. Regular HA use may have a protective effect on cognitive impairment in those with moderate hearing loss.

Understanding the relationships among aging, cognitive function, and olfaction may be useful for diagnosing olfactory decline in older adults. Olfactory function declines in the early stage of neurodegenerative diseases, including Alzheimer's and Parkinson's diseases. Aging and cognitive impairment are associated with olfactory decline. Moreover, the assessment of hyposmia and anosmia is paramount to the diagnosis of neurodegenerative diseases. We aimed to assess the relationships among aging, cognitive function, and olfaction in patients with olfactory impairment.This observational study included 141 patients with olfactory deterioration who presented with dementia, mild cognitive impairment (MCI), age-related hyposmia, or postviral olfactory dysfunction (PVOD). The patients underwent TT olfactometry, a self-administered odor questionnaire (SAOQ), a visual analog scale (VAS), and a Mini-Mental State Examination.TT odor recognition thresholds decreased with aging (p0.01) and cognitive impairment (p 0.08). The average TT recognition thresholds were 5.1,4.6,4.2, and 3.7 in dementia, MCI, age-related hyposmia, and PVOD, respectively. Moreover, the average differences between the detection and recognition thresholds were 3.7, 2.8, 2.3, and 2.0 in dementia, MCI, age-related hyposmia, and PVOD, respectively. Hyposmia with dementia presented the highest recognition thresholds (p 0.05) and the largest differences between the TT detection and recognition thresholds, compared with age-related hyposmia and PVOD (p 0.05). Hyposmia with dementia had the highest SAOQ and VAS scores compared with the other groups (p 0.05).The possibility of dementia should be investigated in patients with hyposmia, including those with high TT recognition thresholds, a large difference between the TT detection and recognition thresholds, and high SAOQ and VAS scores.

Fucoidan is a hetero-sulfated polysaccharide found in brown algae and has received much attention as an ingredient in functional and health foods. The marine bacterial strain Luteolibacter algae H18 degrades fucoidan from Cladosiphon okamuranus. We purified the fucoidanase from a cell-free extract of L. algae H18, used it to decrease the molecular weight of deacetylated-fucoidan, determined the N-terminal amino acid sequence of the enzyme, and identified the gene involved in the degradation of fucoidan, fct114, in a draft genome sequence of strain H18. The gene product was heterologously produced in Escherichia coli and demonstrated to catalyze the degradation of deacetylated-fucoidan into lower molecular weight fragments. The mass of the gene product Fct114 is 112 kDa (1026 amino acid residues). The general properties of the enzyme were investigated by measuring the amount of reducing ends produced from deacetylated-fucoidan during the reaction. The enzyme was inactive toward fucoidans from other brown seaweed species or toward polysaccharides such as alginic acid, carrageenan, hyaluronic acid, and chondroitin sulfate. The amino acid sequence of Fct114 shared less than 25% identity and had no conserved motifs when compared with previously identified fucoidanases from other marine bacterial strains. These data suggest that Fct114 is a novel polysaccharide-degrading enzyme.

Stress-induced mutagenesis can assist pathogens in generating drug-resistant cells during antibiotic therapy; however, if and how antibiotics induce mutagenesis in microbes remains poorly understood. A non-pathogenic thermophile, Geobacillus kaustophilus HTA426, efficiently produces derivative cells resistant to rifampicin and streptomycin via rpoB and rpsL mutations, respectively. Here, we examined this phenomenon to suggest a novel mutagenic mode induced by antibiotics. Fluctuation analysis indicated that mutations occurred via spontaneous mutations during culture. However, mutations were much more frequent in growing cells than stationary cells, and mutation sites were varied through cell growth. These observations suggested that growing cells induced mutagenesis in response to antibiotics. An in-frame deletion of mfd, which governs transcription-coupled repair to correct DNA lesions on the transcribed strand, caused mutations that were comparable between growing and stationary cells; therefore, the mutagenic mechanism was attributable to DNA repair defects where growing cells depressed mfd function. Mutations occurred more frequently at optimal growth temperatures for G. kaustophilus than at a higher growth temperature, suggesting that the mutagenesis relies on active cellular activities rather than high temperature-associated DNA damage. In addition, the mutagenesis may involve a mutagenic factor targeting these sites, in addition to mfd depression, because rpoB and rpsL mutations were dominant at thymine and guanine sites on the transcribed strand. A similar mutagenic profile was observed for other Geobacillus and thermophilic Bacillus species. This suggests that Bacillus-related thermophiles commonly induce mutagenesis in response to rifampicin and streptomycin to produce resistant cells.

Objective The influence of living alone on multiple food and nutrition behaviors across a range of ages and genders has not been sufficiently investigated. Moreover, to our knowledge, no studies have described both dietary status and medical examination findings in persons living alone. Among individuals who attended a health checkup in a rural town in Japan, we investigated dietary habits and medical examination findings in persons living alone and those living with one or more other persons. Methods The participants in this investigation were 501 community-dwelling individuals aged 40 to 91 y (mean, 63.8 ± 9.9 y). Thirty-four (16.4%) of the 207 men and 45 (15.3%) of the 294 women lived alone. Dietary intake frequency of 28 types of foods and drinks, including various vegetables, fruits, meat, seafood, and dairy products, was investigated based on responses to a self-completed questionnaire. During the physical examination, body weight, body mass index, body fat percentage, and blood pressure (systolic and diastolic) were measured. Blood examination included red blood cell count, white blood cell count, platelet count, hemoglobin, hematocrit, total protein, albumin/globulin ratio, total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, creatinine, blood urea nitrogen, uric acid, and serum calcium. Results Multivariate analysis revealed that living alone independently was associated with a reduced intake frequency of various vegetables and seafood, especially in men. Diastolic blood pressure and triglyceride levels were significantly higher in men living alone than in men living with other persons. The albumin/globulin ratio in women living alone was significantly lower than in women living with other persons. Conclusions It was found that living alone was associated not only with a reduced quality of dietary intake but also with an unfavorable trend in some of the medical examination findings.

Geobacillus spp. are moderate thermophiles that have great potential for use in diverse applications. For effective utilization of the species, genetic tools have been extensively studied; however, an overexpression vector remains to be developed. Here we constructed a plasmid vector that can shuttle between Escherichia coli and Geobacillus spp., and which contained a maltose-inducible promoter from Geobacillus kaustophilus HTA426. Although the vector (termed pGKE119) was originally designed for basal gene expression, it surprisingly directed robust protein production in G. kaustophilus. Protein production essentially occurred in an auto-inducible manner without maltose; however, some proteins were produced more efficiently in the presence of maltose. Although the productivity was affected by culture conditions, three proteins were successfully produced with abundance ratios of 12–27% (on a total protein basis) and yields of 77–170 mg (per L culture). pGKE119 directed substantial protein production even in Geobacillus subterraneus, Geobacillus thermoglucosidasius, and Geobacillus thermoleovorans. This suggests that pGKE119 can use a range of Geobacillus spp. as hosts and widely expand their genetic toolbox. Because Geobacillus spp. are highly proliferative bacteria that are distinct from organisms used as protein production hosts, pGKE119 may also provide a novel platform for hyperproduction of recombinant proteins.

Background and Purpose: In the horizontal canal benign paroxysmal positional vertigo (BPPV), cupulolithiasis shows apogeotropic direction changing nystagmus lasting more than 1 min, while canalolithiasis leads to geotropic direction changing nystagmus lasting < 1 min. The difference between cupulolithiasis and canalolithiasis is widely accepted to be the attachment of the displaced otoconia to the cupula of a semicircular canal. Several studies have shown a relationship between BPPV and vitamin D deficiency, but no studies have compared serum levels of vitamin D between canalolithiasis and cupulolithiasis patients. The purpose of this study was to clarify the difference in vitamin D serum level between canalolithiasis and cupulolithiasis of the horizontal canal. Methods: This retrospective study included 20 and 15 patients with canalolithiasis and cupulolithiasis of the horizontal canal, respectively. Serum levels of 25-hydroxyvitamin D [25(OH)D] during the acute phase of BPPV were measured. Results: The mean 25(OH)D serum level in patients with canalolithiasis and cupulolithiasis was 13.2 ± 1.4 and 20.4 ± 1.6 ng/mL, respectively, and the difference was statistically significant (p = 0.0014), also after adjusting for age and sex (p = 0.0351). Eighteen out of 20 (90%) and 5 of 15 (33%) patients were diagnosed with vitamin D deficiency in the canalolithiasis and cupulolithiasis groups, respectively, and this difference was also statistically significant (p = 0.0005). Conclusion: We found that serum vitamin D level in patients with canalolithiasis was significantly lower than that in patients with cupulolithiasis of the horizontal canal.

Background and objectives Hearing impairment (HI) is a major global health concern. In addition, social networks are important for healthy aging. This study aimed to examine the association between HI and social relationships. Research design and methods This study was conducted by the National Institute for Longevity Sciences as part of its Longitudinal Study of Aging with 1176 Japanese participants aged 60 years or older (mean age 71.0 ± 7.4). The convoy model was used to evaluate participants’ network size. A pure-tone average hearing level (HL) of 0.5, 1, 2, and 4 kHz in the better ear >25 dB HL was defined as HI. Multivariate analysis was performed to assess the relationship between HI and the network size, adjusting for age, gender, years of education, presence of depressive symptoms, and higher-level functional capacity score. Results The mean network sizes across the three circles of the convoy model differed significantly by HI status (18.7 ± 0.4 in the no-HI group vs 17.0 ± 0.5 in the HI group, p = 0.003). In particular, the number of non-kin in the outer circle was significantly less in the HI group (4.1 ± 0.2 vs 3.3 ± 0.3, p = 0.004). Discussion and implications The social network size was significantly smaller in the HI group. The outer circle of people to whom the individual feels less close and the number of non-kin were related to the presence of HI. Therefore, HI may be associated with elderly people’s social relationships.

The aims of this study were to determine the prevalence of MRI abnormalities which were suspected as sinusitis in community-dwelling middle-aged and elderly Japanese and to identify risk factors for the MRI abnormality. Brain magnetic resonance imaging (MRI) data from the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA) were used for the analysis. Among the 2330 subjects in the NILS-LSA, 1933 participants were categorized as having no MRI abnormality or MRI abnormality using the Lund-Mackay (LM) score. The mean LM score of the participants was 0.88±1.92, and 144 (7.4%) participants had MRI abnormalities which were suspected as sinusitis when it was classified as an LM score greater than or equal to 4. The prevalence of MRI abnormality was significantly higher in participants of older age and the male sex, in participants with obesity, hypertension, bronchial asthma, chronic bronchitis, gout, or hyperuricemia and in ex- or current smokers. A multivariate logistic regression revealed that older age (odds ratio [OR] = 1.17), obesity (OR = 1.54), a smoking habit (OR = 1.71), history of asthma (OR = 3.77), and chronic bronchitis (OR = 2.66) were significant risk factors for MRI abnormality.

The plasmid pGKE75- cat A138T , which comprises pUC18 and the cat A138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CAT A138T ), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75- cat A138T in an error-prone thermophile, generating the mutant plasmid pGKE75 αβ - cat A138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75 αβ - cat A138T contained no mutation in the cat A138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus . Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75 αβ - cat A138T is attributable to increases in intracellular CAT A138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75 αβ - cat A138T . The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus . It is noteworthy that G. kaustophilus was transformed with pGKE75 αβ - cat A138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli .

Thermostability is an important property of enzymes utilized for practical applications because it allows long-term storage and use as catalysts. In this study, we constructed an error-prone strain of the thermophile Geobacillus kaustophilus HTA426 and investigated thermoadaptation-directed enzyme evolution using the strain. A mutation frequency assay using the antibiotics rifampin and streptomycin revealed that G. kaustophilus had substantially higher mutability than Escherichia coli and Bacillus subtilis . The predominant mutations in G. kaustophilus were A · T→G · C and C · G→T · A transitions, implying that the high mutability of G. kaustophilus was attributable in part to high-temperature-associated DNA damage during growth. Among the genes that may be involved in DNA repair in G. kaustophilus , deletions of the mutSL , mutY , ung , and mfd genes markedly enhanced mutability. These genes were subsequently deleted to construct an error-prone thermophile that showed much higher (700- to 9,000-fold) mutability than the parent strain. The error-prone strain was auxotrophic for uracil owing to the fact that the strain was deficient in the intrinsic pyrF gene. Although the strain harboring Bacillus subtilis pyrF was also essentially auxotrophic, cells became prototrophic after 2 days of culture under uracil starvation, generating B. subtilis PyrF variants with an enhanced half-denaturation temperature of >10°C. These data suggest that this error-prone strain is a promising host for thermoadaptation-directed evolution to generate thermostable variants from thermolabile enzymes.

Fucoidan is an α-l-fucopyranosyl polymer found in seaweeds with forms that have acetyl and sulfuric modifications and derivatives that are lower and/or diversified, with modifications that have attracted interest as potential bioactive substances. We identified the gene for a fucoidan deacetylase that cleaves acetyl moieties from fucoidan and thereby contributes to fucoidan utilization in the marine bacterium Luteolibacter algae H18. Fucoidan deacetylase was purified to homogeneity from a cell-free extract of L. algae H18, and used to determine the internal amino acid sequence and identify the gene, fud, in a draft genome sequence of the H18 strain. The gene product was heterologously produced in Escherichia coli and was demonstrated to catalyze fucoidan deacetylation, but not desulfation, and degradation into lower forms. In addition to fucoidan deacetylation, the enzyme catalyzed the hydrolysis of p-nitrophenyl esters with organic acids, and p-nitrophenyl acetate was the best substrate among those tested. The present study provides a new tool for fucoidan degradation, potentially expanding investigations on fucoidan derivatives.

In medical and pharmacological research, various human disease models in small fish, such as medaka (Oryzias latipes), have been created. To investigate these disease models noninvasively, magnetic resonance imaging (MRI) is suitable because these small fish are no longer transparent as adults. However, their small body size requires a high spatial resolution and a water pool should be avoided to maximize the strength of MRI. We developed in vivo magnetic resonance microscopy (MR microscopy) without a water pool by combining hypothermic anaesthesia and a 14.1 T MR microscope. Using in vivo MR microscopy, we noninvasively evaluated the hepatic steatosis level of a non-alcoholic fatty liver disease model in medaka and followed the individual disease progression. The steatosis level was quantified by the MRI-estimated proton density fat-fraction (MRI-PDFF), which estimates the triglyceride fat concentration in liver tissue and is recognized as an imaging biomarker. The MRI-PDFF results agreed with a histological analysis. Moreover, we optimized the hypothermic anaesthesia procedure to obtain a recovery proportion of 1 in the experiment involving MR microscopy. Recovered medaka could not be distinguished from naïve medaka after the experiment. Therefore, the in vivo MR microscopy will expand the possibilities of a human disease model in fish.

The etiologies of idiopathic sudden sensorineural hearing loss (SSNHL) and Ménière's disease remain unclear. Recently, accumulating evidence has demonstrated that oxidative stress is related to the pathology of inner ear disease. Because genetic factors may contribute partly to the etiologies of SSNHL and Ménière's disease, we investigated the associations between genetic polymorphisms located in oxidative stress response genes and susceptibility to SSNHL and Ménière's disease. We compared 84 patients affected by SSNHL, 82 patients affected by Ménière's disease, and 2107 adults (1056 men and 1051 women; mean age, 59.2 years; range, 40-79 years) who participated in the National Institute for Longevity Sciences, Longitudinal Study of Aging. Multiple logistic regression was used to calculate odds ratios for SSNHL and Ménière's disease in individuals with polymorphisms in the genes glutathione peroxidase 1 (GPX1) (Pro198Leu, rs1050450), paraoxonase 1 (PON1) (Gln192Arg, rs662; and Met55Leu, rs854560), PON2 (Ser311Cys, rs7493), and superoxide dismutase 2 (SOD2) (Val16Ala, rs4880), with adjustment for age and gender. No significant differences in the distribution of the genotypes at these polymorphisms were observed among individuals with SSNHL and Ménière's disease and controls. No significant risk for SSNHL and Ménière's disease was observed in the additive genetic model, regardless of moderating variables. The C allele of SOD2 (rs4880) was more frequent in Ménière's disease cases with a hearing level over 50 dB compared with cases with a hearing level below 50 dB, suggesting that this polymorphism is associated with progression of a hearing loss in Ménière's disease. In conclusion, no significant associations between the polymorphisms of GPX1, PON1, PON2, and SOD2 and risk of SSNHL and Ménière's disease were observed in this Japanese case-control study.

We established an efficient transformation method for thermophile Geobacillus kaustophilus HTA426 using conjugative transfer from Escherichia coli of host-mimicking plasmids that imitate DNA methylation of strain HTA426 to circumvent its DNA restriction barriers. Two conjugative plasmids, pSTE33T and pUCG18T, capable of shuttling between E. coli and Geobacillus spp., were constructed. The plasmids were first introduced into E. coli BR408, which expressed one inherent DNA methylase gene (dam) and two heterologous methylase genes from strain HTA426 (GK1380-GK1381 and GK0343-GK0344). The plasmids were then directly transferred from E. coli cells to strain HTA426 by conjugative transfer using pUB307 or pRK2013 as a helper plasmid. pUCG18T was introduced very efficiently (transfer efficiency, 10(-5)-10(-3) recipient(-1)). pSTE33T showed lower efficiency (10(-7)-10(-6) recipient(-1)) but had a high copy number and high segregational stability. Methylase genes in the donor substantially affected the transfer efficiency, demonstrating that the host-mimicking strategy contributes to efficient transformation. The transformation method, along with the two distinguishing plasmids, increases the potential of G. kaustophilus HTA426 as a thermophilic host to be used in various applications and as a model for biological studies of this genus. Our results also demonstrate that conjugative transfer is a promising approach for introducing exogenous DNA into thermophiles.

Genipin, a herbal iridoid, is known to have both neuroprotective and neuritogenic activity in neuronal cell lines. As it is structurally similar to tetrahydrobiopterin, its activity is believed to be nitric oxide (NO)-dependent. We previously proposed a novel neuroprotective activity of a genipin derivative, (1R)-isoPropyloxygenipin (IPRG001), whereby it reduces oxidative stress in RGC-5, a neuronal precursor cell line of retinal origin through protein S-nitrosylation. In the present study, we investigated another neuritogenic property of IPRG001 in RGC-5 cells and retinal explant culture where in we focused on the NO-cGMP-dependent and protein S-nitrosylation pathways. IPRG001 stimulated neurite outgrowth in RGC-5 cells and retinal explant culture through NO-dependent signaling, but not NO-dependent cGMP signaling. Neurite outgrowth with IPRG001 requires retinoic acid receptor β (RARβ) expression, which is suppressed by an RAR blocking agent and siRNA inhibition. Thereby, we hypothesized that RARβ expression is mediated by protein S-nitrosylation. S-nitrosylation of histone deacetylase 2 is a key mechanism in chromatin remodeling leading to transcriptional gene activation. We found a parallelism between S-nitrosylation of histone diacetylase 2 and the induction of RARβ expression with IPRG001 treatment. The both neuroprotective and neuritogenic activities of genipin could be a new target for the regeneration of retinal ganglion cells after glaucomatous conditions.

Conclusion. Imaging of the endolymphatic space was possible in most cases after administering a standard dose of gadolinium (Gd) by using heavily T2-weighted 3D fluid-attenuated inversion recovery (hT(2)W-3D-FLAIR) magnetic resonance imaging (MRI). Objectives. The aim of this study was to characterize the contrast enhancement in the inner ear after intravenous injection of Gd and to compare the signal intensity after a double dose or standard dose of Gd. Methods: We injected a standard dose of gadodiamide hydrate (DTPA-BMA) or a double dose of gadoteridol (HP-DO3A) intravenously in patients affected by Meniere's disease and performed hT(2)W-3D-FLAIR 3 T MRI. Contrast enhancement of the basal turn of the cochlea was evaluated semiquantitatively. Results: Endolymphatic hydrops was observed in the cochlea and vestibule of the affected ears of patients with Meniere's disease in both the double-dose group and the standard-dose group.

Introduction: IgM composes a part of the B cell receptor, and its expression is restricted to B cell lineage cells. There have been several reports that refractory B cell malignant cells frequently express IgM. For example, activated B cell-like (ABC) diffuse large cell lymphoma (DLBCL) is a subtype with a poor prognosis following R-CHOP treatment (Lenz et al., N Engl J Med, 2008), and the tumor cells generally express IgM-type immunoglobulin (Ruminy et al., Leukemia, 2011). In addition, IgM-expressing chronic lymphocytic leukemia (CLL) was also found to have a poor prognosis (D'Avola et al., Blood, 2016). Therefore, IgM is a promising therapeutic target for refractory B cell malignancies. However, anti-IgM antibodies may not function fully in the body due to the presence of soluble IgM. To improve the selectivity to the IgM expressing cells, we produced a bispecific antibody bound to IgM and HLA-DR, which are both expressed by B cells, and it was able to bind IgM+HLA-DR+ cells in the presence of soluble IgM. Thus, the aim of the present study was to examine the in vitro anti-tumor activity of the anti-IgM/HLA-DR bispecific antibody against B cell lymphoma, and to clarify its efficacy and tolerability in non-human primates (NHP). Methods: BTA106 is the bispecific antibody that can bind to IgM and HLA-DR, and BTA111 is glycoengineered BTA106. To investigate their direct inhibitory effects and Fc-mediated effects against B cell lymphoma cell lines, a growth inhibition assay, antibody-dependent cell cytotoxicity (ADCC) assay and complement dependent cytotoxicity (CDC) assay were performed. In the growth inhibition assay and CDC assay, cell proliferation was measured by WST-8. In the ADCC assay, human peripheral blood mononuclear cells were used as effector cells. Dead cells were stained with propidium iodide and measured by flow cytometry. JeKo-1 (Mantle cell lymphoma, MCL), Ramos (Burkitt's lymphoma, BL), RL (DLBCL), B104 (B cell lymphoma) and two rituximab-resistant cell lines, RL-4RH (Czuczman et al., Clin Cancer Res., 2008) and RRBL1 (generously provided by Dr. Tomita, Tomita et al., Int J Hematol., 2007), were used. To clarify the efficacy and tolerability of BTA106 in NHP, a dose escalation study and single-dose administration study were performed using cynomolgus monkeys. Results: BTA106 and BTA111 demonstrated direct growth inhibition activity, and CDC and ADCC activity, although their intensities differed among cell lines. In the case of rituximab-resistant cell lines, RRBL1 and RL-4RH, BTA106 and BTA111 exhibited weak or no growth inhibitory effects, but their CDC and ADCC activity was superior to that of rituximab. Development of resistance to rituximab is also a poor prognostic factor and remains problematic. Thus, BTA106 and BTA111 are expected as therapeutic antibodies to overcome rituximab resistance. A dose escalation study and single dose study were performed in cynomolgus monkeys. BTA106 greatly reduced peripheral B cells. Moreover, after the administration of BTA106, lymph follicles disappeared in axillary lymph nodes. In these studies, decreased locomotor activity and vomiting were observed; however, these symptoms resolved 2 hrs after injection. Other BTA106-related clinical toxicities or changes in body weight were not observed. Conclusions: In the in vitro studies, BTA106 and BTA111 demonstrated significant anti-tumor effects on rituximab-sensitive and -resistant B cell lymphoma cell lines. The efficacy and toxicity studies in cynomolgus monkeys revealed that BTA106 depletes peripheral and lymph node B cells, and it was well tolerated. BTA111 was considered to be significantly more effective than BTA106 due to glycoengineering, which induced high ADCC activity. These data suggest that BTA111 is an effective and potent therapeutic antibody for B cell malignancies, especially rituximab-resistant B cell lymphoma. Currently, a repeated administration study of BTA111 with cynomolgus monkeys to evaluate its efficacy and toxicity is ongoing. Disclosures Ohashi: Zenyaku Kogyo Co., Ltd.: Employment. Miyashita:Zenyaku Kogyo Co., Ltd.: Employment. Nagata:Zenyaku Kogyo Co., Ltd.: Employment. Tatebe:Zenyaku Kogyo Co., Ltd.: Employment. Otsuka:Zenyaku Kogyo Co., Ltd.: Employment. Suzuki:Zenyaku Kogyo Co., Ltd.: Employment. Sasaki:Zenyaku Kogyo Co., Ltd.: Employment. Enami:Zenyaku Kogyo Co., Ltd.: Employment. Tsukada:Zenyaku Kogyo Co., Ltd.: Employment. Fukushima:Zenyaku Kogyo Co., Ltd.: Employment.

The efficacy of aerosolization of ciprofloxacin (CPFX) incorporated into PEGylated liposomes (PEGylated CPFX-liposomes) for the treatment of respiratory infections was evaluated.PEGylated CPFX-liposomes with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-n-[methoxy(polyethylene glycol)-2000] (particle size: 100 nm) were prepared, and the drug distribution characteristics in lung epithelial lining fluid (ELF) following aerosolization of PEGylated CPFX-liposomes were examined in rats. Furthermore, the antibacterial effects of PEGylated CPFX-liposomes in ELF were evaluated by pharmacokinetic/pharmacodynamic analysis.The elimination rate of CPFX from ELF following aerosolization of PEGylated CPFX-liposomes was significantly slower than that of CPFX incorporated into unmodified liposomes (unmodified CPFX-liposomes; particle size: 100 nm). According to pharmacokinetic/pharmacodynamic analysis, the PEGylated CPFX-liposomes exhibited potent antibacterial effects against pathogenic microorganisms in ELF.This study shows that PEGylated CPFX-liposomes are a useful aerosol-based pulmonary drug delivery system for the treatment of respiratory infections.

The generation of space charges in polymeric dielectric materials is known to be affected by such inclusions as cross-linking by-products, antioxidants, and moisture. We reported previously that negative heterocharges in XLPE are generated in the presence of acetophenone and sulfur-containing antioxidants. In this paper we describe further experiments using samples containing DCP and three different antioxidants in order to examine the mechanism of this phenomenon. Negative heterocharges near the positive electrode were detected in samples containing both DCP and antioxidants when those samples were soaked in acetophenone. Negative heterocharges were not detected in samples containing either DCP or the antioxidant, even when the sample was soaked in acetophenone. These results demonstrate that a heterocharge is generated by the combined effect of acetophenone and the substance produced by interaction between DCP and the antioxidant. Based on the results of experiments measuring the amount of heterocharge in samples that had been heatpressed at different temperatures, the mechanism of negative heterocharge generation in polymeric dielectric material is inferred. © 2010 Wiley Periodicals, Inc. Electr Eng Jpn, 172(2): 1–9, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/eej.20941

Previously, we reported that genipin, a herbal iridoid, had neuritogenic and neuroprotective actions on PC12 cells. Although nitric oxide (NO)-activated signalings were proposed to be neuritogenic, the neuroprotective action of genipin remains to be elucidated. From the standpoint of NO activation, we tested a possible protective mechanism through the nitrosative Kelch-like ECH-associated protein (Keap1)/NF-E2-related factor 2 (Nrf2)-antioxidant response element pathway in rat retinal ganglion cells (RGC-5 cells) in culture, and in vivo, against hydrogen peroxide and optic nerve injury (ONI), respectively, using a long-acting (1R)-isoPropyloxygenipin (IPRG001). IPRG001 induced NO generation and the expressions of antioxidative enzymes, such as heme oxygenase-1 (HO-1), in RGC-5 cells. The protective action of IPRG001 depended on HO-1 and NO induction. We found that S-nitrosylation of Keap1 by IPRG001 may contribute to translocation of Nrf2 to the nucleus and triggered transcriptional activation of antioxidative enzymes. Furthermore, apoptotic cells were increased and 4-hydroxy-2-nonenal was accumulated in rat retina following ONI. Pre-treatment with IPRG001 almost completely suppressed apoptosis and accumulation of 4-hydroxy-2-nonenal in RGCs following ONI accompanied by HO-1 induction. These data demonstrate for the first time that IPRG001 exerts neuroprotective action in RGCs in vitro and in vivo, through the Nrf2/antioxidant response element pathway by S-nitrosylation against oxidative stress.

We designed 1-alkyloxygenipins with the aim of improving the stability of genipins based on the structural and electronic properties of genipins, and prepared 1-alkyloxygenipins and examined their neuritogenic activities in PC12h cells. All genipin-derivatives exhibited electronic properties similar to those of genipin and induced significant neurite outgrowth. These compounds will be classified as nitric oxide synthase (NOS) activators (neuritogenic active compounds) since their lowest unoccupied molecular orbital (LUMO)-energies are similar to that of tetrahydrobiopterin (H4B). (1R)-isoPropyloxygenipin showed activity comparable to that of genipin, and unlike the parent compound genipin, it was found to be physiologically stable in rat liver homogenate.

Background Minimally invasive endoscopic fetal surgery enables intrauterine intervention with reduced risk to the mother and fetus. A novel surgical manipulator is described for stabilizing the fetus and restraining it from floating free during endoscopic intrauterine surgery. Methods We designed and fabricated a prototype fetus-supporting manipulator equipped with flexible joint and bending mechanisms and a soft balloon stabilizer. The flexible joint and bending mechanisms enable the stabilizer to reach the target sites within the confined space of the uterus under the guidance of an ultrasound device. The balloon stabilizer could be inserted into the uterus through a small incision. Results The accuracy evaluation showed that the maximum error of the bending mechanism was as small as 7 mm and the standard deviation of the joint mechanism was just 1.6 ◦ . In the experiments using a fetus model, the manipulator could be well controlled with guidance from ultrasound images and its bending mechanism with the balloon stabilizer could be clearly visualized while stabilizing the fetus model. Conclusions The manipulator has the potential to be used in minimally invasive intrauterine surgery, although further improvements and experiments remain to be carried out. Copyright  2008 John Wiley & Sons, Ltd.

The present investigation was undertaken to compare the structural and electronic properties of genipin with those of tetrahydrobiopterin (H4B), an essential cofactor in neuronal nitric oxide synthase (nNOS), and 4-aminotetrahydrobiopterin (4-amino-H4B), an inhibitor of nNOS, using computer-assisted molecular modeling techniques. Molecular modeling, superimposing, and docking simulation, in addition to LUMO-energy calculation, revealed that genipin has structural and electronic properties that markedly resemble those of H4B.

We previously demonstrated efficient transformation of the thermophile Geobacillus kaustophilus HTA426 using conjugative plasmid transfer from Escherichia coli BR408. To evaluate the versatility of this approach to thermophile transformation, this study examined genetic transformation of various thermophilic Bacillus and Geobacillus spp. using conjugative plasmid transfer from E. coli strains. E. coli BR408 successfully transferred the E. coli-Geobacillus shuttle plasmid pUCG18T to 16 of 18 thermophiles with transformation efficiencies between 4.1 × 10(-7) and 3.8 × 10(-2)/recipient. Other E. coli strains that are different from E. coli BR408 in intracellular DNA methylation also generated transformants from 9 to 15 of the 18 thermophiles, including one that E. coli BR408 could not transform, although the transformation efficiencies of these strains were generally lower than those of E. coli BR408. The conjugation was performed by simple incubation of an E. coli donor and a thermophile recipient without optimization of experimental conditions. Moreover, thermophile transformants were distinguished from abundant E. coli donor only by high temperature incubation. These observations suggest that conjugative plasmid transfer, particularly using E. coli BR408, is a facile and versatile approach for plasmid introduction into thermophilic Bacillus and Geobacillus spp., and potentially a variety of other thermophiles.

Based on our results regarding the structure-activity relationships of alkylxanthines and imidazo[2,1-i]purines as phosphodiesterase 4 (PDE4) inhibitors, we designed new 1-benzylxanthines and investigated their PDE4 inhibitory activities. 3,7-Dihydro-7-acetonyl-1-(2,4-dichlorobenzyl)-3-propyl-1H-purine-2,4-dione (2h) exhibited both more selective and more potent PDE4 inhibitory activity than rolipram and XT-611.

Microbes employ cell membranes for reducing exogenous stresses. Polyamino acid display on microbial cell surfaces and their effects on microbial chemical stress tolerance were examined. Growth analysis revealed that displays of polyarginine, polyaspartate and polytryptophan substantially enhanced tolerance of Escherichia coli to NaCl. A titration assay indicated that polyarginine and polyaspartate altered cell surface charges, implying tolerance enhancement via ion atmosphere and/or ionic bond network formations for electrostatic ion repulsion. The enhancement by polytryptophan may have arisen from surface hydrophobicity increase for hydrophobic ion exclusion, because of a strong correlation between hydrophobic characters of amino acids and their effects on tolerance enhancement. The display also enhanced tolerance to other salts and/or alcohols in E. coli and to NaCl in Saccharomyces cerevisiae. Thus polyamino acid display has the potential as an approach for conferring chemical stress tolerance on various microbes.

To establish a methodology for magnetic resonance imaging (MRI) assessment in the diagnosis of cholesteatoma using signal intensity on BLADE diffusion-weighted MRI (BLADE-DWI) and apparent diffusion coefficient (ADC) mapping.Retrospective case series.University hospital.Participants comprised 29 patients who underwent middle ear surgery and in whom preoperative differential diagnosis between cholesteatoma and other middle ear diseases was difficult using local and computed tomographic findings and required BLADE-DWI.Signal intensity ratio (SIR) between the affected region of the middle ear and the pons measured by BLADE-DWI and on ADC maps was evaluated numerically. SIR in an area located near the target lesion in each case was used as a control. Values were compared between both cases in which cholesteatoma was histopathologically confirmed (cholesteatoma group) and cases in which cholesteatoma was excluded on histopathologic examination (noncholesteatoma group).Imaging and histopathologic findings.SIR on BLADE-DWI was significantly higher in the cholesteatoma group than in the noncholesteatoma group, although both groups showed significantly higher SIR in the target lesion than in the control area. Moreover, SIR on ADC maps was significantly lower in the cholesteatoma group than in the noncholesteatoma group. The clear cutoff value of SIR on ADC maps was 1.5.The combination of BLADE-DWI and ADC mapping offers a useful imaging tool for accurate detection of middle ear cholesteatoma. Use of SIR can numerically differentiate between cholesteatoma and noncholesteatoma.

Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression of enzymatic activities in vivo . To capitalize on these advantages of thermophiles, we describe here a new inducible gene expression system in the thermophile Geobacillus kaustophilus HTA426. Six promoter regions in the HTA426 genome were identified and analyzed for expression profiles using β-galactosidase reporter assay. This analysis identified a promoter region upstream of a putative amylose-metabolizing gene cluster that directed high-level expression of the reporter gene. The expression was >280-fold that without a promoter and was further enhanced 12-fold by maltose addition. In association with a multicopy plasmid, this promoter region was used to express heterologous genes. Several genes, including a gene whose product was insoluble when expressed in Escherichia coli , were successfully expressed as soluble proteins, with yields of 0.16 to 59 mg/liter, and conferred new functions to G. kaustophilus strains. Remarkably, cellulase and α-amylase genes conferred the ability to degrade cellulose paper and insoluble starch at high temperatures, respectively, generating thermophiles with the potential to degrade plant biomass. Our results demonstrate that this novel expression system expands the potential applications of G. kaustophilus .

Objectives/Hypothesis Endothelin-1 is a potent vasoconstrictor peptide that is widely distributed throughout the mammalian body including the spiral modiolar artery, vestibule, and cochlea. This study aimed to investigate the association between the Lys198Asn (G/T) polymorphism (rs5370) of the endothelin-1 gene and sudden sensorineural hearing loss (SSNHL). Study Design Case-control study. Methods Seventy-two SSNHL patients (mean age, 58.3 ± 14.0 years) were compared with 2,159 controls included in a community-based study of aging. Multiple logistic regression was used to obtain odds ratios (ORs) for SSNHL. In subgroup analysis, patients with SSNHL who visited to the hospital within the first month of onset were selected to assess audiometric features according to genotype. Pure-tone averages at 250, 500, 1,000, 2,000, and 4,000 Hz were calculated in the affected ear. Results Under the recessive genetic model, after adjustment for age, sex, histories of hypertension, dyslipidemia and diabetes, the crude and adjusted ORs for SSNHL risk were 2.209 (95% confidence interval [CI]: 1.140-4.281) and 2.173 (95% CI: 1.086-4.348), respectively. No significant ORs were observed under the additive and dominant models. The severity of SSNHL differed significantly between genotypes. The mean pure-tone averages at the initial visit were 78.6, 66.4, and 57.8 dB for the GG, GT, and TT genotypes, respectively (P = .034). Conclusions Our study indicates that the recessive genotype was significantly associated with increased SSNHL risk; however, the severity was lower in these individuals than it was in those with the wild-type genotype. Endothelin-1 may be implicated in SSNHL. Level of Evidence 3b Laryngoscope, 123:E59–E65, 2013

The etiologies of idiopathic sudden sensorineural hearing loss (SSNHL) and Meniere's disease remain unclear. Recently, accumulating evidence has demonstrated that free radicals are related to the pathology of inner ear disease. Because genetic factors may contribute partly to the etiologies of SSNHL and Meniere's disease, we investigated the association between genetic polymorphisms located in genes related to the free-radical process and susceptibility to SSNHL and Meniere's disease. We compared 83 patients affected by SSNHL and 83 patients affected by Meniere's disease with 2048 adults (for SSNHL) and 1946 adults (for Meniere's disease) who participated in the National Institute for Longevity Sciences, Longitudinal Study of Aging. Multiple logistic regression was used to calculate odds ratios (ORs) for SSNHL and Meniere's disease in individuals with polymorphisms in the genes: methionine synthase (MTR; rs1805087); methionine-synthase reductase (MTRR; rs1801394); nitric oxide synthase 3 (NOS3; rs1799983); caveolin 1 (Cav1; rs3840634); melatonin receptor 1B (MTNR1B; rs1387153); NAD(P)H oxidase p22(phox) subunit (NADH/NADPHp22phox; rs4673); and mitochondria 5178 (MT5178; rs28357984). The NOS3 polymorphism was significantly associated with a risk of SSNHL; in addition, the OR for the NOS3 polymorphism and SSNHL risk was 2.108 (CI, 1.343-3.309) with adjustment for age and sex. The Cav1 polymorphism was significantly associated with a risk of Meniere's disease; moreover, the OR for the Cav1 polymorphism and Meniere's disease risk was 1.849 (CI, 1.033-3.310) with adjustment for age and sex. In conclusion, the NOS3 and Cav1 polymorphisms were significantly associated with the risk of SSNHL and Meniere's disease, respectively.

1,3,5-Triazacycloheptanes were synthesized and examined for reversal of the multidrug resistance dependent on P-glycoprotein. Most of these compounds increased the intracellular uptake of vinblastine in multidrug-resistant mouse leukemia P388/ADR cells without influence upon the vinblastine accumulation in P388/S cells. The efficacy of 1,5-dibenzyl-1,3,5-triazacycloheptanes in increasing the vinblastine accumulation was in the order of 2,4-dithioxo (5) > 2-oxo-4-thioxo (4) approximately 4-(methylthio)-2-oxo (6) > 2,4-dioxo (2). The efficacy was further increased when the benzyl group was converted to a chlorobenzyl group. Among these compounds, 6c [1,5-bis(4-chlorobenzyl)-1,5,6,7-terahydro-4-(methylthio)-2H-1,3,5 - triazepin-2-one] potentiated the in vitro cell growth-inhibitory effect of vinblastine, adriamycin, and mitomycin C on P388/ADR cells and prolonged the life span of P388/ADR-bearing mice in combined therapy with vinblastine more than vinblastine alone.

Urinary albumin fragments (uAF) from patients with NIDDM were analyzed as a possible factor in the early discovery of diabetic nephropathy before emergence of microalbuminuria. SDS-PAGE and immunoblot assay were employed for detection of uAF. Samples from 252 patients with NIDDM, 158 patients with non-diabetic diseases, and 48 healthy volunteers were examined; uAF were detected in 139 (55.2%), 94 (59.5%), only one (2.1%), respectively. In diabetic patients with normoalbuminuria, uAF were detected in 48 out of 159 cases (30.2%). Two years after the initial study, 3 of the 17 diabetic patients (17.6%) with normoalbuminuria and uAF showed micro- or macroalbuminuria. It was concluded that detection of uAF might be useful for the early detection of diabetic nephropathy in NIDDM.

In 1928, Griffith [1] reported that soluble substances from virulent pneumococcal cells trans‐ formed non-virulent pneumococcus to virulent forms. This substance has now been demon‐ strated to be DNA [2-4]. This is considered to be the first report on genetic transformation of bacteria by exogenous DNA. Subsequently, natural competence of Bacillus subtilis was re‐ ported in 1958 by Young and Spizizen [5]. They also demonstrated genetic transformation of natural competent B. subtilis cells using exogenous DNA. It was in 1970 that genetic trans‐ formation of Escherichia coli using chemically competent cells was reported [6]. Thus, genetic transformation of common bacterial models was established at an early stage in the develop‐ ment of bacteriology. The alternative view is that bacterial models such as B. subtilis and E. coli have become the mainstay of this field because of high transformation ability. Genetic transformation techniques remain important for studying numerous bacteria and for the ad‐ vancement of bacteriology, biochemistry, applied microbiology, and microbial biotechnolo‐ gy. Moreover, recent developments in the search for new bacteria and genome sequencing have provided numerous effective bacteria that are useful for biological studies and indus‐ trial applications. With these developments, there is a greater demand for establishing ge‐ netic transformation methods for more bacteria.

Although the etiology of idiopathic sudden sensorineural hearing loss (SSNHL) remains unclear, the pathologically increased permeability of blood vessels, elucidated by gadolinium-enhanced magnetic resonance imaging (MRI), suggests the involvement of inflammation. Because SSNHL is considered a multifactorial disease, possibly caused by interactions between genetic factors and environmental factors, the authors investigated the associations of polymorphisms of inflammatory mediator genes with susceptibility to SSNHL. The authors compared 72 patients affected by SSNHL and 2010 adults (1010 men and 1000 women; mean age 59.2 years; range 40-79) who participated in the National Institute for Longevity Sciences Longitudinal Study of Aging. Multiple logistic regression was used to obtain odds ratios (ORs) for SSNHL in subjects with polymorphisms in the genes IL-6 C - 572G, IL-4R G1902A, IL-10 A - 592C, TNFα C - 863A, TNFRSF1B G593A, VEGF C936T, VEGF C - 2578A, and VEGF G - 1154A, with adjustment for age, gender, and any history of hypertension, diabetes, or dyslipidemia. The per-allele OR for the risk of SSNHL in subjects bearing IL-6 C - 572G was 1.480 (95% confidence interval [CI], 1.037-2.111) in model 1 (no adjustment), 1.463 (CI, 1.022-2.094) in model 2 (adjusted for age and gender), and 1.460 (CI, 1.016-2.097) in model 3 (adjusted for age, gender, and a history of hypertension, diabetes, or dyslipidemia). Under the dominant model of inheritance, the ORs were 1.734 (CI, 1.080-2.783) in model 1, 1.690 (CI, 1.050-2.721) in model 2, and 1.669 (CI, 1.035-2.692) in model 3. The remaining seven polymorphisms failed to show any associations with the risk of SSNHL. These data need to be confirmed on larger series of patients. In conclusion, the IL-6 C - 572G polymorphism is associated with a risk of SSNHL. Because permeability of blood vessels in the inner ear is frequently increased in patients with SSNHL, inflammation of the inner ear might be involved.

A DNA fragment of 42 kb encompassing one of the mycinamicin II (Mm)-resistance-encoding genes, myrB, from a Mm-producing strain, Micromonospora griseorubida, was cloned in Escherichia coli using the cosmid vector pJB8. Nucleotide sequencing of the neighboring region of myrB and a computer-aided analysis of the sequence predicted the presence of an open reading frame (ORF) with 254 amino acids which showed great similarity to the macrocin O-methyltransferase (tylF gene product) in tylosin (Ty)-producing Streptomyces fradiae. When a 1.0-kb AluI fragment containing the complete ORF was fused to the lacZ promoter in the correct orientation and expressed in E. coli, a mycinamicin III (MIII) O-methyltransferase (MOMT) activity was detected only upon induction with isopropyl-beta-D-thiogalactopyranoside (IPTG). All these data indicate that this ORF codes for the structural gene of MOMT, and it is designated mycF.