Manganese in toenails is associated with hearing loss at high frequencies in humans

Purpose: Elevated hearing thresholds from high frequencies are known to be one of the hallmarks of age-related hearing loss. Our recent study showed accumulation of manganese (Mn) in inner ears resulting in acceleration of age-related hearing loss in mice orally exposed to Mn. However, there is no evidence showing an association between Mn in non-invasive biological samples and hearing loss in humans evaluated by pure tone audiometry (PTA). In this study, we evaluated Mn in non-invasive biological samples as a possible biomarker for hearing loss in humans.

Materials and methods: We determined hearing levels by PTA and Mn levels in toenails, hair and urine with an inductively coupled plasma mass spectrometer (ICP-MS) in 145 healthy subjects in Bangladesh.

Results: Multivariable analyses showed that Mn levels in toenails, but not in hair and urine samples, were significantly associated with hearing loss at 8?kHz and 12?kHz. Moreover, our experimental study showed a significant correlation between Mn levels in inner ears and nails, but not hair, in mice orally exposed to Mn.

Conclusions: The results provide novel evidence that Mn in toenails is a possible biomarker for hearing loss at high frequencies in humans.     

Identification of a novel series of potent and selective CCR6 inhibitors as biological probes

CCR6 has been implicated in both autoimmune diseases and non-autoimmune diseases. Thus, inhibition of CCR6-dependent cell migration is an attractive strategy for their treatment. An orally available small molecule inhibitor of CCR6 could therefore be a useful biological probe for the pathophysiological studies. Initial SAR study of a hit compound provided potent N-benzenesulfonylpiperidine derivatives that suppressed CCL20-induced Gi signals. By subsequent scaffold morphing of the central ring and further optimization, we identified a novel series of 1,4-trans-1-benzenesulfonyl-4-aminocyclohexanes as potent and selective CCR6 inhibitors with good pharmacokinetic properties. Our compounds showed good correlation between Gi signal inhibitory activity and cell migration inhibitory activity in human CCR6-transfected CHO cells. In addition, representative compound 35 potently inhibited CCR6-dependent cell migration and the increase in ERK phosphorylation in human primary cells. Therefore, the compound could be used effectively as a biological probe against human CCR6.     

Transcription Factor IRF8 Governs Enhancer Landscape Dynamics in Mononuclear Phagocyte Progenitors

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Amplification of PFGE-separated chromosome-specific DNA by degenerate oligonucleotide-primed PCR (DOP-PCR)

Specific probes for DNA molecules of high molecular weight can be efficiently and rapidly prepared by the combination of pulsed-field gel electrophoresis (PFGE) with degenerate oligonucleotide-primed PCR (DOP-PCR). More than 10 mg of a specific probe were amplified from only 1 ng of a PFGE-separated DNA fragment of a red alga, Cyanidioschyzon merolae. The probe hybridized specifically to the fragment which was used as a template for DOP-PCR.     

Tissue Distribution, Genomic Structure, and Chromosome Mapping of Mouse and Human Eukaryotic Initiation Factor 4E-Binding Proteins 1 and 2

Two related eukaryotic initiation factor-4E binding proteins (4E-BP1 and 4E-BP2) were recently characterized for their capacity to bind specifically to eIF4E and inhibit its function. Here, we determined the cDNA sequence, tissue distribution, genomic structure, and chromosome localization of murine and human 4E-BP1 and 4E-BP2. Mouse 4E-BP1 and 4E-BP2 consist of 117 and 120 amino acids and exhibit 91.5 and 95.0% identity, respectively, to their human homologues. 4E-BP1 mRNA is expressed in most tissues, but is most abundant in adipose tissue, pancreas, and skeletal muscle, while 4E-BP2 mRNA is ubiquitously expressed. The structures of the mouse 4E-BP1 and 4E-BP2 were determined. The 4E-BP1 gene consists of three exons and spans ∼16 kb. In addition, two 4E-BP1 pseudogenes exist in the mouse genome. The 4E-BP2 gene spans approximately 20 kb and exhibits an identical genomic organization to that of 4E-BP1, with the protein coding portion of the gene divided into three exons. There are no pseudogenes for 4E-BP2. The chromosomal locations of 4E-BP1 and 4E-BP2 were determined in both mice and humans by fluorescencein situhybridization analysis. Mouse 4E-BP1 and 4E-BP2 map to chromosomes 8 (A4-B1) and 10 (B4-B5), respectively, and human 4E-BP1 and 4E-BP2 localize to chromosomes 8p12 and 10q21–q22, respectively.     

Production of 5,8,11-Eicosatrienoic Acid (Mead Acid) by a (Delta)6 Desaturation Activity-Enhanced Mutant Derived from a (Delta)12 Desaturase-Defective Mutant of an Arachidonic Acid-Producing Fungus, Mortierella alpina 1S-4

Enhanced production of 5,8,11-eicosatrienoic acid (Mead acid, 20:3(omega)9) was attained by a mutant fungus, Mortierella alpina M209-7, derived from (Delta)12 desaturase-defective M. alpina Mut48. The 20:3(omega)9 production by M209-7 was 1.3 times greater than that by its parent strain, Mut48. This is thought to be due to its enhanced (Delta)6 desaturation activity, which was 1.4 times higher than that of Mut48. In both strains, 87 to 88% of the total lipids comprised triacylglycerol (TG) and 85% of 20:3(omega)9 was contained in TG. On optimization of the culture conditions for M209-7, earlier glucose feeding and shifting of the growth temperature from 28 to 19(deg)C on the second day were shown to be effective. Under the optimal conditions with a 10-liter jar fermentor, 20:3(omega)9 production reached 1.65 g/liter of culture medium (corresponding to 118 mg/g of dry mycelia and 28.9% of total fatty acids), which is about twice that reported previously (0.8 g/liter).     

Approach to a metallacycling polymerization — reactions of [(η-C5H5)Co(PPh3)2] with α,β-diynes and Me3SiCCC6H4C6H4CCSiMe3

Reactions of [CpCo(PPh₃)₂](Cp=η⁵-cyclopentadienyl) with conjugated diacetylenes were investigated in terms of the synthesis of π-conjugated organometallic polymers. The reaction of an α,β-diyne, PhCC---CCPh, gave three geometric isomers of dialkynylcobaltacyclopentadienes, 1a-c, and an insoluble polymeric product, 1d. A 2,4-dialkynyl complex, 2, and a 2,5-dialkynyl complex, 3, were obtained solely from Me₃SiCC---CCSiMe₃ and MeCC---CCMe, respectively. 1,1′-Bis(trimethylsilylethynyl)-4,4′-biphenyl afforded two isomers of 1,3-dialkynylcyclobutadiene complexes, 4a and 4b. The stability of the one-electron oxidized forms of the cobalacyclopentadiene and cyclobutadiene complexes was examined by cyclic voltammetry.     

Phospholipids of membranes of cultured cells and the products of protoplast fusion

The variety and distribution of phospholipids in the cell membranes of cultured cells and their fatty acid composition were analysed. Membranes of suspension cultured cells of Rauwolfia serpentina var. Bentham, Nicotiana tabacum var. Samsun, Atropa belladonna and Bouvardia ternifolia had almost the same composition of phosphatidylcholine, PC (ca 50%); phosphatidylethanolamine, PE (ca 25%); phosphatidylinositol, PI (ca 10%); phosphatidylglycerol, PG (several %); and phosphatidic acid, PA (several %). We determined the distribution of the molecular species of the three major phospholipid fractions (PC, PE and PI) in R.serpentina and N.tabacum. Membranes of both cell-types contained basically similar molecular species, 1–16:0/2–18:2 the main type, particularly in the PC- and PE- fractions; 1–18:2/2–18:2 and 1–18:0/2–18:2 for R. serpentina; and 1–16:0/2–18:3, 1-18:0/2-18:3 and 1-18:2/2-18:2 for N.tabacum. The influence of membrane fluidity on protoplast fusion as effected by the phase transition of the phospholipids, is discussed.     

Effects of concanavalin A on the net formation of zoospores in Hydrodictyon reticulatum (Chlorococcales, Chlorophyceae)

In the green alga Hydrodictyon reticulatum (L.) Lager‐heim (Chlorococcales, Chlorophyceae), zoospores are arranged in a regular fashion to form an intricate hexagonal network during the asexual reproductive cycle. Polypeptides that bind concanavalin A (Con A) in zoospores increased in amount during net formation and decreased after the completion of the adhesion between zoospores. Fluorescein isothiocyanate‐Con A‐binding sites corresponded to the contact sites of zoospores immediately after cessation of the movement. Treatment with 25 μg mL^?1 Con A inhibited adhesion of isolated immotile zoospores obtained from parental cells, and Con A‐treated zoospores could not form hexagonal nets. Moreover, when isolated immotile zoospores were treated with Con A, the cessation of the zoospore movement was retarded in dose‐dependent manner. These results suggest that the Con A‐binding sites may participate in the adhesion of zoospores during hexagonal net formation.