Missense Mutation in the Alternative Splice Region of the PAX6 Gene in Eye Anomalies

SummaryThe PAX6 gene is involved in ocular morphogenesis, and PAX6 mutations have been detected in various types of ocular anomalies, including aniridia, Peters anomaly, corneal dystrophy, congenital cataract, and foveal hypoplasia. The gene encodes a transcriptional regulator that recognizes target genes through its paired-type DNA-binding domain. The paired domain is composed of two distinct DNA-binding subdomains, the N-terminal subdomain (NTS) and the C-terminal subdomain (CTS), which bind respective consensus DNA sequences. The human PAX6 gene produces two alternative splice isoforms that have the distinct structure of the paired domain. The insertion, into the NTS, of 14 additional amino acids encoded by exon 5a abolishes the DNA-binding activity of the NTS and unmasks the DNA-binding ability of the CTS. Thus, exon 5a appears to function as a molecular switch that specifies target genes. We ascertained a novel missense mutation in four pedigrees with Peters anomaly, congenital cataract, Axenfeldt anomaly, and/or foveal hypoplasia, which, to our knowledge, is the first mutation identified in the splice-variant region. A T→A transition at the 20th nucleotide position of exon 5a results in a Val→Asp (GTC→GAC) substitution at the 7th codon of the alternative splice region. Functional analyses demonstrated that the V54D mutation slightly increased NTS binding and decreased CTS transactivation activity to almost half

THOREAU ト EMERSON ノ ルイジ ト イシツ シゼン ト シャカイ ニツイテ

In this paper, I attempted to clarify similarities and differences as seen between Thoreau and Emerson with special reference to their common theme, that is, nature and society. Apparently they were not only contemporary transcendentalists but also had an enormous amount of their own writings and journals. But their most substantial similarity can be located in their search for nature;that is, they tried to find the universal existence behind matter by placing themselves in harmony with nature. On the one hand, by analogizing the final correspondence between nature and spirit, Emerson could attain the ideation by means of matter. Because he used matter as symbols of his passion, he succeeded in conforming things to his thoughts so that he might integrate nature, God and man. On the other hand, although Thoreau had a firm belief in existence of divinity in man, he failed to find the ideation by means of matter. However, Thoreau recognized the various phases of nature more definitely than Emerson. Eventuallyj came to the conclusion that the central difference between their thoughts lay in a way of searching nature. Taking critical attitudes toward conservatism prevailing in the then society, Thoreau and Emerson insisted on individualism, that is, the greater im-portance of the individual than society. Both of them keenly reacted to the contemporary society. Owing to his Conviction of the boundless progress of the individual and the notion of scientific progress, Emerson could not only consider evil as something transient, but also accept the realities of society such as the State, the state government, and the Negro slavery. On the other hand, Thoreau could never accept the existing governrnent, the State and the Negro slavery involving the social evil as a civil government, and explicated his rebellious mind based on his conscience and justice of the individual against the formalism in his society. Finally, Emerson took an affirmative attitude toward the American society of which he tried to forsee the future. For Thoreau\u27s own partas he could never overlook any social evil, he seriously took part in the antislavery movement. Gathering these facts together, I could make it clear that Thoreau\u27s views about society was more radical than Emerson\u27s. Furthermore, Thoreau persisted in his self-reliant attitude, and he tried hard to carry it more deeply in his everyday life than Emerson

A Three-Dimensional FRET Analysis to Construct an Atomic Model of the Actin–Tropomyosin–Troponin Core Domain Complex on a Muscle Thin Filament

It is essential to knowthe detailed structure of the thin filament to understand the regulation mechanism of striated muscle contraction. Fluorescence resonance energy transfer (FRET) was used to construct an atomic model of the actin–tropomyosin (Tm)–troponin (Tn) core domain complex. We generated single-cysteine mutants in the 167–195 region of Tm and in TnC, TnI, and the β-TnT 25-kDa fragment, and each was attached with an energy donor probe. An energy acceptor probe was located at actin Gln41, actin Cys374, or the actin nucleotide-binding site. From these donor–acceptor pairs, FRET efficiencies were determined with and without Ca2+. Using the atomic coordinates for F-actin, Tm, and the Tn core domain, we searched all possible arrangements for Tm or the Tn core domain on F-actin to calculate the FRET efficiency for each donor–acceptor pair in each arrangement. By minimizing the squared sum of deviations for the calculated FRET efficiencies from the observed FRET efficiencies, we determined the location of Tm segment 167– 195 and the Tn core domain on F-actin with andwithout Ca2+. The bulk of the Tn core domain is located near actin subdomains 3 and 4. The central helix of TnC is nearly perpendicular to the F-actin axis, directing the N-terminal domain of TnC toward the actin outer domain. The C-terminal region in the I–T arm forms a four-helix-bundle structure with the Tm 175–185 region. After Ca2+ release, the Tn core domainmoves toward the actin outer domain and closer to the center of the F-actin axis

Novel urinary glycan profiling by lectin array serves as the biomarkers for predicting renal prognosis in patients with IgA nephropathy

In IgA nephropathy (IgAN), IgA1 molecules are characterized by galactose deficiency in O-glycans. Here, we investigated the association between urinary glycosylation profile measured by 45 lectins at baseline and renal prognosis in 142 patients with IgAN. The primary outcome was estimated glomerular filtration rate (eGFR) decline (>4 mL/min/1.73 m(2)/year), or eGFR >= 30% decline from baseline, or initiation of renal replacement therapies within 3 years. During follow-up (3.4 years, median), 26 patients reached the renal outcome (Group P), while 116 patients were with good renal outcome (Group G). Multivariate logistic regression analyses revealed that lectin binding signals of Erythrina cristagalli lectin (ECA) (odds ratio [OR] 2.84, 95% confidence interval [CI] 1.11-7.28) and Narcissus pseudonarcissus lectin (NPA) (OR 2.32, 95% CI 1.11-4.85) adjusted by age, sex, eGFR, and urinary protein were significantly associated with the outcome, and they recognize Gal(beta 1-4)GlcNAc and high-mannose including Man(alpha 1-6)Man, respectively. The addition of two lectin-binding glycan signals to the interstitial fibrosis/tubular atrophy score further improved the model fitness (Akaike's information criterion) and incremental predictive abilities (c-index, net reclassification improvement, and integrated discrimination improvement). Urinary N-glycan profiling by lectin array is useful in the prediction of IgAN prognosis, since ECA and NPA recognize the intermediate glycans during N-glycosylation of various glycoproteins