Expansion of GGC Repeat in GIPC1 Is Associated with Oculopharyngodistal Myopathy.

Oculopharyngodistal myopathy (OPDM) is an adult-onset inherited neuromuscular disorder characterized by progressive ptosis, external ophthalmoplegia, and weakness of the masseter, facial, pharyngeal, and distal limb muscles. The myopathological features are presence of rimmed vacuoles (RVs) in the muscle fibers and myopathic changes of differing severity. Inheritance is variable, with either putative autosomal-dominant or autosomal-recessive pattern. Here, using a comprehensive strategy combining whole-genome sequencing (WGS), long-read whole-genome sequencing (LRS), linkage analysis, repeat-primed polymerase chain reaction (RP-PCR), and fluorescence amplicon length analysis polymerase chain reaction (AL-PCR), we identified an abnormal GGC repeat expansion in the 5' UTR of GIPC1 in one out of four families and three sporadic case subjects from a Chinese OPDM cohort. Expanded GGC repeats were further confirmed as the cause of OPDM in an additional 2 out of 4 families and 6 out of 13 sporadic Chinese individuals with OPDM, as well as 7 out of 194 unrelated Japanese individuals with OPDM. Methylation, qRT-PCR, and western blot analysis indicated that GIPC1 mRNA levels were increased while protein levels were unaltered in OPDM-affected individuals. RNA sequencing indicated p53 signaling, vascular smooth muscle contraction, ubiquitin-mediated proteolysis, and ribosome pathways were involved in the pathogenic mechanisms of OPDM-affected individuals with GGC repeat expansion in GIPC1. This study provides further evidence that OPDM is associated with GGC repeat expansions in distinct genes and highly suggests that expanded GGC repeat units are essential in the pathogenesis of OPDM, regardless of the genes in which the expanded repeats are located.

Intranuclear inclusions in skin biopsies are not limited to neuronal intranuclear inclusion disease but can also be seen in oculopharyngodistal myopathy.

AIMS: Oculopharyngodistal myopathy (OPDM) is caused by the expansion of CGG repeats in NOTCH2NLC (OPDM_NOTCH2NLC) GIPC1 (OPDM_GIPC1), or LRP12 (OPDM_LRP12). Neuronal intranuclear inclusion disease (NIID) is clinically distinct from OPDM but is also caused by the expansion of CGG repeats in NOTCH2NLC, which may be an indicator of intranuclear inclusion in skin biopsy. We investigated the presence of intranuclear inclusions in skin biopsies from patients with OPDM and muscle diseases with a similar pathology to evaluate whether they will have similar diagnostic findings on skin biopsy. METHODS: We analysed the frequency of p62-positive intranuclear inclusions in sweat gland cells, adipocytes and fibroblasts in skin biopsy samples from patients with OPDM (OPDM_NOTCH2NLC [n = 2], OPDM_GIPC1 [n = 6] and OPDM_LRP12 [n = 3]), NIID (n = 1), OPMD (n = 1), IBM (n = 4) and GNE myopathy (n = 2). RESULTS: The p62-postive intranuclear inclusions were observed in all three cell types in both patients with OPDM_NOTCH2NLC and a patient with NIID, in at least one cell type in all six patients with OPDM_GIPC1, and all in three cell types in one of the three patients with OPDM_LRP12. These findings were not observed in patients with OPMD, IBM or GNE myopathy. CONCLUSION: Intranuclear inclusions in skin biopsy samples are not specific to NIID and are found in all three types of genetically confirmed OPDM, suggesting that the underlying mechanism of OPDM may be similar to NIID, regardless of causative genes.

Real-time high-spectral-resolution mid-infrared spectroscopy with a signal-to-noise ratio of ten thousand.

We developed a mid-infrared spectroscopy system with high spectral resolution and a high signal-to-noise ratio using an extremely high-order germanium immersion grating. The spectroscopic system covers wavelengths from 3 to 5 µm and has a spectral resolution of 1 GHz with a single-shot bandwidth of 2 THz. We proposed a method of improving the signal-to-noise ratio and achieved a ratio of over 3000 with a data acquisition rate of 125 Hz in the presence of fluctuations in the light source and environment. A signal-to-noise ratio of 10,000 was achieved with 0.1-s integration for 100-µW mid-infrared light.

Stability of the transamidase complex catalyzing GPI anchoring of proteins.

Glycosylphosphatidylinositol (GPI) is a glycolipid that anchors some proteins to the plasma membrane. This anchoring is catalyzed by a transamidase complex (TAC) composed of five subunits: PIG-K, GAA1, PIG-U, PIG-T, and PIG-S (Fig. 1A). PIG-K and GAA1 are predicted to catalyze the first and second steps during attachment of proproteins of GPI-anchored proteins (GPI-APs) to GPI. GPI may be delivered by PIG-U, and PIG-T is required for stability of all TAC subunits when overexpressed in cultured cells. However, protein stability of TAC has not been analyzed using loss-of-function mutants for each subunit. Herein, we analyzed the stability of TAC in knockout and/or knockdown mutants for each subunit. PIG-T and PIG-U, or PIG-T and GAA1, were mutually required for stability, and all three subunits were stable without PIG-S or PIG-K. However, these three subunits were essential for the stability of both PIG-S and PIG-K. By contrast, loss of PIG-S reduced the stability of PIG-K and left the other subunits unaffected. Reduction of PIG-K did not impact any of the other subunits. Thus, PIG-T, PIG-U, and GAA1 may form a core complex associated by PIG-S, and these four subunits may stabilize PIG-K, triggering GPI anchoring reactions. Instability of PIG-K in the absence of the other four subunits may ensure that GPI anchoring is catalyzed only by the completely assembled complex.

Microwave discharge electrodeless lamps (MDEL). Part VII. Photo-isomerization of trans-urocanic acid in aqueous media driven by UV light from a novel Hg-free Dewar-like microwave discharge thermally-insulated electrodeless lamp (MDTIEL). Performance evaluation.

A novel mercury-free Dewar-like (double-walled structure) microwave discharge thermally-insulated electrodeless lamp (MDTIEL) was fabricated and its performance evaluated using the photo-isomerization of trans-urocanic acid (trans-UA) in aqueous media as a test process driven by the emitted UV light when ignited with microwave radiation. The photo-isomerization processes trans-UA → cis-UA and cis-UA → trans-UA were re-visited using light emitted from a conventional high-pressure Hg light source and examined for the influence of UV light irradiance and solution temperature; the temperature dependence of the trans → cis process displayed a negative activation energy, E(a) = -1.3 cal mol(-1). To control the photo-isomerization of urocanic acid from the heat usually dissipated by a microwave discharge electrodeless lamp (single-walled MDEL), it was necessary to suppress the microwave-initiated heat. For comparison, the gas-fill in the MDEL lamp, which typically consists of a mixture of Hg and Ar, was changed to the more eco-friendly N(2) gas in the novel MDTIEL device. The dynamics of the photo-isomerization of urocanic acid driven by the UV wavelengths of the N(2)-MDTIEL light source were compared to those from the more conventional single-walled N(2)-MDEL and Hg/Ar-MDEL light sources, and with those from the Hg lamp used to irradiate, via a fiber optic, the photoreactor located in the wave-guide of the microwave apparatus. The heating efficiency of a solution with the double-walled N(2)-MDTIEL was compared to the efficiency from the single-walled N(2)-MDEL device. Advantages of N(2)-MDTIEL are described from a comparison of the dynamics of the trans-UA → cis-UA process on the basis of unit surface area of the lamp and unit power consumption. The considerably lower temperature on the external surface of the N(2)-MDTIEL light source should make it attractive in carrying out photochemical reactions that may be heat-sensitive such as the photothermochromic urocanic acid system.

[A case of liver metastasis from gastric cancer responding to S-1/CDDP chemotherapy, allowing partial gastrectomy and left hepatectomy].

A 63-year-old man was found to have an upper abdominal mass, and was referred to our hospital. Endoscopic examination and abdominal CT showed gastric cancer with liver metastasis. A large metastatic tumor of the liver invaded the hepatic hilus, making curative resection impossible. We started chemotherapy using S-1(120 mg/body/day), orally administered for three weeks followed by 2-week rest period, and cisplatin(100mg/body), administered intravenously on day 8 as 1 course. After 5 courses of chemotherapy, the liver tumor reduced markedly and the gastric cancer pathologically disappeared, enabling partial gastrectomy and left hepatectomy. Histological examination showed a well-differentiated adenocarcinoma in the mucosal layer of the resected stomach. A resected specimen of the liver showed a moderately-differentiated adenocarcinoma with signet-ring cells, compatible to liver metastasis from gastric cancer. Bile leakage the remaining liver occurred, but he recovered soon. Gastrointestinal examination revealed another early gastric cancer after seeing him for 2 years on an outpatient basis. We conducted subtotal gastrectomy, and the patient remains alive 30 months after the first operation. This case suggests that S-1/CDDP chemotherapy may reduce the stage of unresectable liver metastasis from gastric cancer and make a curative operation possible.

Rapid synthesis of Gemini surfactants using a novel 915-MHz microwave apparatus.

Rapid synthesis of Gemini surfactants (C(12)-C(2)-C(12) and C(14)-C(6)-C(14)) by microwave heating is investigated. The yield of the synthesis of C(12)-C(2)-C(12) surfactant using 2.45-GHz microwaves was twice the yield obtained by the oil bath method. Moreover, the value of dielectric loss and microwave penetration depth for the sample solution suggest that the microwave frequency of 915 MHz (0.915 GHz) is preferable over the conventional frequency (2.45 GHz). A novel 915-MHz microwave organic synthesis apparatus with a closed reactor is proposed. The synthesis yields of C(12)-C(2)-C(12) obtained using the 915-MHz equipment were three to four times higher than those obtained using the conventional heating method.

Regulation of transcript levels of the Arabidopsis cytochrome p450 genes involved in brassinosteroid biosynthesis.

Cytochrome P450 enzymes of the closely related CYP90 and CYP85 families catalyze essential oxidative reactions in the biosynthesis of brassinosteroid (BR) hormones. Arabidopsis CYP90B1/DWF4 and CYP90A1/CPD are responsible for respective C-22 and C-23 hydroxylation of the steroid side chain and CYP85A1 catalyzes C-6 oxidation of 6-deoxo intermediates, whereas the functions of CYP90C1/ROT3, CYP90D1, and CYP85A2 are still unknown. Semiquantitative reverse transcriptase-polymerase chain reaction analyses show that transcript levels of CYP85 and CYP90 genes are down-regulated by brassinolide, the end product of the BR biosynthesis pathway. Feedback control of the CYP90C1, CYP90D1, and CYP85A2 genes by brassinolide suggests that the corresponding enzymes might also participate in BR synthesis. CYP85 and CYP90 mRNAs show strong and transient accumulation during the 1st week of seedling development, as well as characteristic organ-specific distribution. Transcripts of CYP90A1 and CYP85A2 are preferentially represented in shoots and CYP90C1, CYP90D1, and CYP85A1 mRNAs are more abundant in roots, whereas CYP90B1 is ubiquitously expressed. Remarkably, the spatial pattern of CYP90A1 expression is maintained in the BR-insensitive cbb2 mutant, indicating the independence of organ-specific and BR-dependent regulation. Quantitative gas chromatography-mass spectrometry analysis of endogenous BRs in shoots and roots of Arabidopsis, pea (Pisum sativum), and tomato (Lycopersicon esculentum) reveal similar partitioning patterns of BR intermediates in these species. Inverse correlation between CYP90A1/CPD transcript levels and the amounts of the CYP90A1 substrate 6-deoxocathasterone in shoots and roots suggests that transcriptional regulation plays an important role in controlling BR biosynthesis.