Flying characterization of colliding partners by hyperfine splitting frequency of neutral paramagnetic atoms.

The pseudopotentials and dispersion potentials are applied to a theoretical study of the hyperfine splitting frequencies of the ground-state paramagnetic hydrogen (H) and alkali-metal (Li, Na, K, Rb, and Cs) atoms in noble gases (He, Ne, Ar, Kr, and Xe). Using classical turning points for statistical averages, we find that numerical calculations based on second-order perturbation theory fit the measured frequency shifts well over a wide temperature range. The characteristic energy, pseudopotential height, and electric-dipole polarizability allow us to consistently determine the van der Waals radii and electron scattering lengths of noble-gas atoms. This study shows that the hyperfine splitting frequency of alkali-metal atoms is a good measure for investigating colliding partners.

Pseudopotential analysis on hyperfine splitting frequency shift of alkali-metal atoms in noble gases, revisited.

Theoretical pseudopotentials and dispersion potentials are used to study ground-state hyperfine splitting frequencies of alkali-metal atoms (Li, Na, K, Rb, and Cs) in noble gases (He, Ne, Ar, Kr, and Xe) in all combinations. With a single fitting parameter, calculations based on first-order perturbation theory qualitatively present each temperature dependence of the measured frequency shift. With this parameter and excitation energies of alkali-metal and noble-gas atoms, the hyperfine splitting frequency of alkali-metal atoms is suitable for investigating the properties of noble-gas atoms, such as the s-wave scattering length of electrons, the electric-dipole polarizability, and the van der Waals radius. This study suggests the possibility of improving excitation energies and van der Waals potentials of colliding pairs.

[Cyclic thrombocytopenia diagnosed by periodic platelet count measurements].

Cyclic thrombocytopenia (CTP) is characterized by periodic platelet count fluctuations and is commonly misdiagnosed as immune thrombocytopenia (ITP) because of their similar clinical characteristics. Here, we present the case of a 74-year-old man with CTP diagnosed by weekly platelet count measurements. The patient initially developed mild bleeding symptoms with a platelet count of 0.8×104/µl. Bone marrow biopsy exhibited reduced megakaryocyte counts, an atypical characteristic of ITP. Weekly follow-up of platelet counts demonstrated an apparent cyclic pattern, resulting in CTP diagnosis. Regular platelet count measurements can help diagnose CTP in thrombocytopenia.

Nephronectin influences EAE development by regulating the Th17/Treg balance via reactive oxygen species.

Blood levels of the extracellular matrix protein nephronectin (Npnt), a protein critical for kidney development, are elevated in autoimmune experimental autoimmune encephalitis (EAE) mice, which are a model for multiple sclerosis. We found here that treatment with anti-Npnt antibody directed against the α8ß1 integrin-binding site (Npnt-blocking antibody) inhibits EAE development. The selenium transporter selenoprotein P (SeP) was identified as a novel Npnt-binding partner. In EAE, Npnt induced SeP and glutathione peroxidase 1 (GPx1) expression, followed by reactive oxygen species (ROS) inhibition in CD4+ T cells; these changes were disturbed by Npnt-blocking antibody treatment, which also caused suppressed differentiation of interleukin (IL)-17-producing CD4+ T-helper cells (Th17s) and elevated differentiation of regulatory T cells (Tregs). Treatment of EAE mice with the ROS scavenger N-acetyl cysteine (NAC) blocked the Npnt-blocking antibody-induced decrease in Th17 differentiation and increase in Treg differentiation. In conclusion, the interaction between Npnt and SeP contributes to EAE development by regulating the Th17/Treg balance via the ROS level.

Noble-gas atoms characterized by hyperfine frequency shift of lithium atom.

We report an experimental and theoretical study on the shift of the hyperfine splitting frequency of ground-state Li atoms in noble gases, He, Ne, Ar, and Xe. The frequency shift is due to the change in the electron-spin density at the Li nuclei induced by collisions to the noble-gas atoms. The electron density is calculated along the interatomic distance in a pseudopotential and a dispersion potential. Based on the measured and the calculated frequency shifts, we find the importance of attractive force in collisions to helium as well as heavy noble-gas atoms. Taking advantage of the simple energy structure of the Li atom, we obtain the s wave scattering length for free electrons on noble-gas atoms by using the hyperfine splitting frequency as a precise measure.

Involvement of natriuretic peptide system in C2C12 myocytes.

The natriuretic peptide system, a key regulator of cGMP signaling, comprises three types of natriuretic peptides, osteocrin/musclin (OSTN), and their natriuretic peptide receptors. Although this system plays important roles in many organs, its physiological roles in skeletal muscle have not been clearly described. In the present study, we investigated the role of the natriuretic peptide system in C2C12 myocytes. All three natriuretic peptide receptors were expressed by cells differentiating from myoblasts to myotubes, and natriuretic peptide receptor B (NPR-B) transcripts were detected at the highest levels. Further, higher levels of cGMP were generated in response to stimulation with C-type natriuretic peptide (CNP) versus atrial natriuretic peptide (ANP), which reflected receptor expression levels. A cGMP analog downregulated the expression of a few ER stress-related genes. Furthermore, OSTN gene expression was strongly upregulated after 20 days of differentiation. Augmented gene expression was found to correlate closely with endoplasmic reticulum (ER) stress, and C/EBP [CCAAT-enhancer-binding protein] homologous protein (CHOP), which is known to be activated by ER stress, affected the expression of OSTN. Together, these results suggest a role for natriuretic peptide signaling in the ER stress response of myocytes.

Spectroscopic study of a diffusion-bonded sapphire cell for hot metal vapors.

Characteristics of a diffusion-bonded sapphire cell for optical experiments with hot metal vapors were investigated. The sapphire cell consisted of sapphire-crystal plates and a borosilicate-glass tube, which were bonded to each other by diffusion bonding without any binders or glues. The glass tube was attached to a vacuum manifold using the standard method applied in glass processing, filled with a small amount of Rb metal by chasing with a torch, and then sealed. The cell was baked at high temperatures, and optical experiments were then performed using rubidium atoms at room temperature. The sapphire cell was found to be vacuum tight, at least up to 350°C, and the sapphire walls remained clear over all temperatures. From the optical experiments, the generation of a background gas was indicated after baking at 200°C. The background gas pressure was low enough to avoid pressure broadening of absorption lines but high enough to cause velocity-changing collisions of Rb atoms. The generated gas pressure decreased at higher temperatures, probably due to chemical reactions.

Expression of a truncated form of the endoplasmic reticulum chaperone protein, σ1 receptor, promotes mitochondrial energy depletion and apoptosis.

The σ1 receptor (σ(1)R) regulates endoplasmic reticulum (ER)/mitochondrial interorganellar Ca(2+) mobilization through the inositol 1,4,5-trisphosphate receptor (IP(3)R). Here, we observed that expression of a novel splice variant of σ(1)R, termed short form σ(1)R (σ(1)SR), has a detrimental effect on mitochondrial energy production and cell survival. σ(1)SR mRNA lacks 47 ribonucleotides encoding exon 2, resulting in a frameshift and formation of a truncated receptor. σ(1)SR localizes primarily in the ER at perinuclear regions and forms a complex with σ(1)R but not with IP(3)R in the mitochondrion-associated ER membrane. Overexpression of both σ(1)R and the truncated isoform promotes mitochondrial elongation with increased ER mitochondrial contact surface. σ(1)R overexpression increases the efficiency of mitochondrial Ca(2+) uptake in response to IP(3)R-driven stimuli, whereas σ(1)SR overexpression reduces it. Most importantly, σ(1)R promotes ATP production via increased mitochondrial Ca(2+) uptake, promoting cell survival in the presence of ER stress. By contrast, σ(1)SR suppresses ATP production following ER stress, enhancing cell death. Taken together, the newly identified σ(1)SR isoform interferes with σ(1)R function relevant to mitochondrial energy production under ER stress conditions, promoting cellular apoptosis.

A Systemically Administered Unconjugated Antisense Oligonucleotide Targeting DUX4 Improves Muscular Injury and Motor Function in FSHD Model Mice.

Facioscapulohumeral muscular dystrophy (FSHD), one of the most common muscular dystrophies, is caused by an abnormal expression of the DUX4 gene in skeletal muscles, resulting in muscle weakness. In this study, we investigated MT-DUX4-ASO, a novel gapmer antisense oligonucleotide (ASO). MT-DUX4-ASO decreased the expression of DUX4 and its target genes in FSHD patient-derived myoblasts. For the first time, we demonstrated that a systemically administered ASO, even without a ligand for drug delivery, could significantly improve muscle injury and motor function in the ACTA1-MCM/FLExDUX4 (DUX4-TG) mouse model of FSHD. Tamoxifen (TMX) injection transiently induces skeletal-muscle-specific DUX4 expression in DUX4-TG mice, while the skeletal muscles of TMX-untreated DUX4-TG mice have leaky DUX4 expression in a small subset of myofibers similar to those of FSHD patients. Subcutaneous 10 mg/kg of MT-DUX4-ASO at two-week intervals significantly suppressed muscular DUX4 target gene expression, histological muscle injury, and blood muscle injury marker elevation in TMX-untreated DUX4-TG mice. Notably, MT-DUX4-ASO at 10 mg/kg every other week significantly prevented the TMX-induced declines in treadmill test running speed and muscle force in DUX4-TG mice. Thus, the systemically administered unconjugated MT-DUX4-ASO suppressed disease progression in DUX4-TG mice, extending the potential of unconjugated ASOs as a promising FSHD treatment strategy.

Antibodies against nephronectin ameliorate anti-type II collagen-induced arthritis in mice.

The extracellular matrix protein nephronectin (Npnt) is known to be critical for kidney development, but its function in inflammatory diseases is unknown. Here, we developed a new enzyme-linked immunosorbent assay system to detect Npnt in various autoimmune diseases, which revealed that plasma Npnt levels are increased in various mouse autoimmune models. We also report that antibodies against the α8ß1 integrin-binding region of Npnt protect mice from anti-type II collagen-induced arthritis, suggesting that Npnt may be a potential therapeutic target molecule for the prevention of autoimmune arthritis.

Establishment of novel mAb to human ERC/mesothelin useful for study and diagnosis of ERC/mesothelin-expressing cancers.

Malignant mesothelioma is a highly aggressive tumor of the serosal cavity that arises from the mesothelial cells of the pleura, peritoneum, or pericardium. The immunohistochemical diagnosis of epithelioid mesothelioma from biopsy or surgically resected specimens has been actively pursued, using markers such as mesothelin. Several markers have indeed been helpful for confirming the diagnosis of mesothelioma and distinguishing between mesothelioma and adenocarcinoma. The authors have developed a novel mAb to human C-ERC/mesothelin, which performed well when used in western blotting, fluorescence-activated cell sorting, immunocytochemistry and immunohistochemistry, and which therefore will be useful in studying the molecular biology of mesothelin, in addition to improving the diagnosis and therapy of mesothelin-expressing cancers.

Simultaneous occurrence of partial anomalous pulmonary venous return and major bronchial anomaly: computed tomography findings in 5 adult patients.

OBJECTIVE: The objective of this study was to describe the computed tomography (CT) findings of 5 adult patients with a combination of partial anomalous pulmonary venous return (PAPVR) and major bronchial anomaly. METHODS: A computerized search of the radiological database records of 4886 consecutive patients who underwent chest CT from April 2005 to May 2007 described 2 patients with a combination of PAPVR and tracheal bronchus (TB). Three additional patients with a combination of PAPVR and TB or accessory cardiac bronchus (ACB) were obtained by reviewing the CT images of the patients initially diagnosed as having PAPVR, TB, or ACB at our institution during the same period. The CT findings of these 5 patients were analyzed. RESULTS: Four patients had a combination of right PAPVR and right TB, and 1 patient had a combination of left PAPVR and ACB. These combinations were found in 42% (5/12) of patients with PAPVR, 16% (4/25) of patients with TB, and 14% (1/7) of patients with ACB. In 4 patients with a combination of right PAPVR and right TB, the anomalous vein was draining the right upper lobe and draining into either the posterior aspect of the superior vena cava (SVC) or the terminal portion of the azygos arch. The origin of the TB was the trachea in 1 patient and the carina in 2 patients. Two distinct TBs presented in 1 patient; the first TB was from the lower trachea, and the second was from the right main bronchus. The right upper lobe affected by PAPVR included the territory ventilated by the TB in all 4 patients. CONCLUSION: A combination of right PAPVR draining to the superior vena cava or azygos arch and right TB is most common. The possibility that the PAPVR and a major bronchial anomaly might coexist in 1 patient should be considered whenever either of them is encountered on CT.

Effect of post exposure bake in inorganic electron beam resist and utilizing for nanoimprint mold.

A high-aspect-ratio structural mold has been fabricated using a simple process that comprises of electron beam lithography (EBL) and post exposure bake (PEB) of inorganic resist. Using developed inorganic resist for mold, pattern transfer to a photo-curable polymer was carried out by ultraviolet nanoimprint lithography (UV-NIL). The developed inorganic resist has enough hardness to be used for mold because this resist structure is almost equivalent to that of quartz. The aspect ratio of the fabricated mold with PEB was 2.59; this value is twice the size of aspect ratio of fabricated mold without PEB (1.16). Thus, PEB is very effective in fabricating high-aspect ratio pattern in inorganic resist. The size of the fabricated mold with PEB was 1004 nm developed depth and 388 nm line and 252 nm space. Using the mold, UV-NIL was carried out. The size of replicated pattern was 942 nm height and 278 nm line and 241 nm space. The replicated height and line width were slightly smaller than mold depth and line width because of volume shrinkage of photo-curable polymer by UV irradiation. The aspect ratio of the replicated pattern using the mold with PEB was 3.39; thus, high-aspect-ratio transfer was also possible. This process is useful for rapid fabrication of quarter wavelength plates.

Morphology-Based Analysis of Myoblasts for Prediction of Myotube Formation.

The development of new drugs depends on the efficiency of drug screening. Phenotype-based screening has attracted interest due to its considerable potency for the discovery of first-in-class drugs. In general, fluorescently labeled imagery is the leading technique for phenotype-based screening; however, there are growing requirements to understand total culture profiles, which are unclear after end-point assays. In this study, we demonstrate that morphology-based cellular evaluation of unlabeled cells is an efficient approach to evaluate myotube formation assays. One of our aims was to study the myogenic differentiation process in C2C12 cells to discern the differences between cellular responses to different medium conditions (serum concentrations and insulin dosages). Our results show that predictive morphological profiles that strongly correlate with myogenic differentiation can be generated from myotube images, even in the confluent stage. The differentiation rate after 14 days can be quantitatively predicted with the highest accuracy by means of images taken on days 0-11.5. In addition, for the application of our morphology-based cellular evaluation, the effect of cyclic guanosine monophosphate (cGMP) on myogenic differentiation was analyzed. Our results show that the quantitated morphological profile from these images can be an effective descriptor for analysis of the myotube-recovering effect of cGMP.

Natriuretic peptide signaling is involved in the expression of oxidative metabolism-related and muscle fiber constitutive genes in the gastrocnemius muscle.

Natriuretic peptides regulate cyclic guanosine monophosphate (cGMP) levels via their receptors and have various physiological effects. Natriuretic peptide receptor C (NPR-C) increases cGMP signaling by functioning as a clearance receptor. We analyzed the role of natriuretic peptides in the skeletal muscle, which increases in mass with bone elongation, of NPR-C- mice. High-fat diet (HFD)-fed NPR-C- mice exhibited obesity resistance and higher oxygen consumption. PGC1α gene expression was upregulated in the gastrocnemius muscle of HFD-fed NPR-C- mice compared with HFD-fed NPR-C+ (wild-type) mice. Gene expression of proliferator-activated receptor delta and estrogen-related receptor α, which upregulate oxidative metabolism, was increased in the gastrocnemius muscle of NPR-C- mice, irrespective of diet. Expression of myosin heavy chain 7, a component of type I slow-twitch fiber, was enhanced. Natriuretic peptide signaling may influence oxidative metabolism-related and slow-twitch fiber constitutive gene expression in the fast-twitch gastrocnemius muscle but not in slow-twitch muscles such as the soleus.

Establishment of a novel specific ELISA system for rat N- and C-ERC/mesothelin. Rat ERC/mesothelin in the body fluids of mice bearing mesothelioma.

Mesothelioma is a type of malignant tumor that most commonly arises from the pleural or peritoneal membrane and is usually associated with previous exposure to asbestos. In humans, ERC/mesothelin is expressed on the normal mesothelium and in some cancers such as mesothelioma or ovarian carcinoma. Recently, several enzyme-linked immunosorbent assay (ELISA) systems for ERC/mesothelin have been developed, the reported usefulness of which has been assessed and demonstrated as a diagnostic tool. However, the basic roles or physiological functions of, and relationship between, ERC/mesothelin and asbestos exposure-mediated carcinogenesis remain to be resolved. In order to elucidate the precise mechanism, animal models of mesothelioma are desperately needed. In this study, we consider the development of a novel specific ELISA system for not only rat N-ERC/mesothelin but also C-ERC/mesothelin, and the first data on the presence of rat ERC/mesothelin in the body fluids of rat malignant mesothelioma-bearing nude mice. The transplanted mice have revealed the higher concentrations of rat N-ERC/mesothelin in the blood and ascites than C-ERC/mesothelin. We hope these novel ELISA systems are useful in the rat model system to clarify the mechanism of asbestos-induced carcinogenesis and to develop new effective drugs for mesothelioma.

MESOMARK kit detects C-ERC/mesothelin, but not SMRP with C-terminus.

ERC/mesothelin is expressed on the normal mesothelium and some cancers such as mesothelioma or ovarian carcinoma. A splicing isoform of ERC/mesothelin (known as SMRP), which has an 82-bp insertion and codes for a C-terminus with a hydrophilic, presumably soluble, tail instead of a GPI-anchoring signal, has been reported as a useful marker for the diagnosis of mesothelioma. However, the existence of SMRP has not yet been demonstrated in the serum of mesothelioma patients. To elucidate the existence of SMRP, we have established a new enzyme-linked immunosorbent assay (ELISA) system for SMRP. The ELISA study revealed that N- and C-ERC/mesothelin were detected in sera from mesothelioma patients, but not SMRP, even in these samples. This result showed that the SMRP detected with MESOMARK kit should be lack of soluble C-terminus and indistinguishable from C-ERC/mesothelin. Further study might be necessary to demonstrate the relationship between SMRP and mesothelin.

Acute effects on the lung and the liver of oral administration of cerium chloride on adult, neonatal and fetal mice.

We evaluated tissue changes associated with cerium chloride administration via gavage to adult mice, via milk to neonatal mice and transplacentally to fetal mice. Change in adults consisted of extensive pulmonary hemorrhage, pulmonary venous congestion, thickened alveolar septae, hepatic necrosis and neutrophil infiltrations. Those in fetal mice consisted of pulmonary and hepatic congestion. These results indicate that gavage cerium administration elicited subtle tissue changes, though oral toxicity is rather low. These changes were less severe in neonatal and fetal mice. When cerium was injected into adult mice through the tail vein, cerium was distributed mainly to the liver, spleen and lung dose-dependently with the cerium concentration gradually decreasing after 3 days. A study of cerium anticoagulation in mouse plasma showed that clotting time was significantly prolonged when cerium was added to plasma. These results suggest that cerium may disturb blood coagulation and cause pulmonary and hepatic vascular congestion.

[A case of long-term survival of postoperative brain metastasis of small cell lung cancer effectively treated with chemotherapy, whole brain radiotherapy and stereotactic radiosurgery].

A 63-year-old woman underwent right upper lobectomy for small cell carcinoma. She received a total of 2 courses of carboplatin and etoposide infusion as adjuvant therapy. One year after the operation, because of elevated serum Pro GRP levels and a metastatic brain tumor revealed by CT, 4 courses of IP therapy (irinotecan 60 mg/m(2), day 1, 8, 15 and cisplatin 60 mg/m(2), day 1, every 4 weeks) and whole brain radiotherapy (2 Gy f, 5 f/week, total 40 Gy) were given. A complete response was obtained, but a tumor relapse occurred ten months after the last chemotherapy. We then performed a stereotactic radiosurgery (marginal dose: 22 Gy, maximum dose 44 Gy), and one month later MRI showed the tumor had shrunk markedly. FDG-PET showed no intensive uptake, suggesting that there was no remaining viable tumor. No severe side effects were observed during these treatments. Currently, the patient has been alive with good performance status and no signs of relapse.

[A case of postoperative multiple pulmonary metastases of non-small-cell lung cancer successfully treated with S-1].

We report a case of postoperative multiple pulmonary metastases of NSCLC successfully treated with S-1. A 72-year-old man underwent rt. lower lobectomy + ND 2 a by VATS. The histopathological examination showed squamous cell carcinoma, pT4 (satellite nodules in same lobe) N0M0. Multiple pulmonary metastases appeared 3 months after operation. We started combination chemotherapy with S-1 and CBDCA. S-1 (100 mg/body) was administered on days 1-21. CBDCA (AUC=5.0) was administered on day 8. Multiple pulmonary metastases almost disappeared after 2 courses of combination chemotherapy. After 6 courses of combination chemotherapy with S-1 and CBDCA, we then converted to S-1 only. Every cycle was repeated every 5 weeks. One year later, there was no sign of disease progression. S-1 is considered to be effective and safely administered in patients with NSCLC.