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.

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.

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.

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.

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.

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.

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.

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.

Analysis of gastrointestinal metastasis of primary lung cancer: Clinical characteristics and prognosis.

The prevalence of gastrointestinal metastasis of lung cancer is low. The aim of the present study was to analyze the frequency and clinical characteristics of metastases to the gastrointestinal tract by retrospectively assessing the clinical records of 2,066 patients with lung cancer. A total of 7 patients (0.33%) were diagnosed with gastrointestinal metastasis, including 4 patients with adenocarcinoma, 1 patient with large cell carcinoma and 2 patients with pleomorphic carcinoma. Furthermore, 3 of the patients presented with small bowel metastases, 2 with gastric metastases, 1 with large bowel metastasis and 1 with metastasis of the appendix. The mean time between the diagnosis of the lung tumors and the identification of gastrointestinal metastasis was 13.5 months (range, 3-49 months). The mean time between the identification of the gastrointestinal metastasis and mortality was 100.6 days (range, 21-145 days). In conclusion, the prognosis of patients with recurrence in distant organs, including the gastrointestinal tract, may be worse than patients with recurrence in distant organs, excluding the gastrointestinal tract, particularly those with symptomatic gastrointestinal metastasis. Therefore, the presence of clinical gastrointestinal metastasis may be life threatening; comprehensive evaluations are required to detect and monitor gastrointestinal metastasis during follow-up.

Repeated Localized Treatment for Endobronchial Metastasis of Thymic Carcinoma.

BACKGROUND: Endobronchial metastases derived from nonpulmonary tumors are uncommon, although a variety of malignant tumors have been reported to be associated with endobronchial metastasis. We herein report a case of repeated bronchoscopic resection of endobronchial metastasis of a thymic carcinoma. CASE REPORT: A 59-year-old woman was diagnosed with primary thymic carcinoma, Masaoka stage IVA, in May 2009. In June 2013, she developed dyspnea. A chest CT scan revealed left upper lobe atelectasis, and a polypoid lesion was noted in the left upper bronchus on bronchoscopy. A pathological examination of the lesion revealed metastatic thymic carcinoma, and bronchoscopic resection was performed for symptom relief. However, the lesion was partially resected, based on the operative findings, which showed the peripheral part of B3 to be the origin of the polypoid lesion and bronchoscopy could not be used to reach this site. Although the patient underwent repeated partial bronchoscopic resection of the polypoid lesion due to the symptoms of dyspnea caused by regrowth of the polypoid metastatic thymic cancer in the left upper bronchus, she remains alive with an excellent performance status and no evidence of widespread or other metastases for more than 5 years after the initial diagnosis. CONCLUSIONS: We speculate that this case was successfully managed with repeated partial bronchoscopic resection because thymic cancer tends to be a slow-growing tumor. Therefore, it is worth resecting endobronchial metastatic thymic carcinoma repeatedly in such cases, even if the resection is partial.

Hyperpolarized cesium ions doped in a glass material.

Hyperpolarized (HP) 133Cs nuclear magnetic resonance signals were measured from borosilicate glass cell walls during optical pumping of cesium vapor at high magnetic field (9.4T). Significant signal enhancements were observed when additional heating of the cell wall was provided by intense but non-resonant laser irradiation, with integrated HP 133Cs NMR signals and line widths varying as a function of heating laser power (and hence glass temperature). Given that virtually no Cs ions would originally be present in the glass, absorbed HP Cs atoms rarely met thermally-polarized Cs ions already at the surface; thus, spin-exchange via nuclear dipole interaction cannot be the primary mechanism for injecting spin polarization into the glass. Instead, it is concluded that the absorption and transport of HP atoms into the glass material itself is the dominant mechanism of nuclear spin injection at high temperatures-the first reported experimental demonstration of such a mechanism.

Long-term survival after surgical treatment of metachronous bilateral adrenal metastases of non-small cell lung carcinoma.

Background Although resection of the metastases is the treatment of choice for unilateral solitary adrenal metastasis of non-small cell lung carcinoma (NSCLC), the surgical treatment for bilateral adrenal metastases is quite rare, likely due to the coexistence of multiple synchronous metastases at other sites and/or primary adrenal insufficiency following bilateral adrenalectomy. We herein report a rare case of asynchronous metastasis of NSCLC to the bilateral adrenal glands with long-term survival after bilateral adrenalectomy. Case Report A 70-year-old male underwent right upper lobectomy for lung adenocarcinomaT2aN2M0, stageIIIA following induction chemotherapy. Forty-four months later, right adrenalectomy of a right adrenal tumor was performed, which revealed metastatic lung carcinoma. Following the administration of adjuvant chemotherapy, a metastatic tumor was detected in the left adrenal gland. Although there were no other signs of distant metastasis on radiological examinations, he underwent the chemotherapy due to the risk of adrenal insufficiency. However, on follow-up CT the adrenal lesion was found to have enlarged; therefore, left adrenalectomy was performed. Three years and six months later, he was doing well, with no evidence of recurrence. Conclusions Selected patients with solitary adrenal metastases of NSCLC can benefit from an aggressive treatment approach, even if such metastases are bilateral.

An analysis of and new risk factors for reexpansion pulmonary edema following spontaneous pneumothorax.

BACKGROUND: The major risk factor for reexpansion pulmonary edema (RPE) following the treatment of spontaneous pneumothorax is thought to be chronic lung collapse. However, a long-term collapsed lung does not always cause RPE. The purpose of this study was to define other risk factors for RPE among patients undergoing drainage for the treatment of spontaneous pneumothorax. METHODS: We retrospectively reviewed all the patients with spontaneous pneumothorax who had been treated at our hospital during a 5-year period. The duration of symptoms, location and size of the pneumothorax, size of the chest tube, and pleural effusion, which can occur coincidentally with pneumothorax, were compared in patients who did and did not experience RPE. RESULTS: Forty patients were underwent drainage for the treatment of a spontaneous pneumothorax between January 2007 and December 2012. RPE developed in 13 of the 40 (32.5%) patients. In the multivariate analysis, the presence of pleural effusion coincident with pneumothorax contributed to the risk for RPE [odds ratios (OR), 1.557; 95% confidence intervals (CI), 1.290-1.880]. The duration of symptoms, location and size of the pneumothorax and size of the chest tube were similar between the groups. Symptomatic RPE was associated with a larger pneumothorax size. CONCLUSIONS: The rate of RPE following spontaneous pneumothorax is higher than was previously reported. Our findings suggest the presence of pleural effusion coincidentally with pneumothorax may therefore be a new risk factor for RPE.

Utility of the serum ProGRP level for follow-up of pulmonary carcinoid tumors.

PATIENT: Female, 67. FINAL DIAGNOSIS: Pulmonary carcinoid tumor. SYMPTOMS: Abnormal shadow on chest X-ray. MEDICATION: -. CLINICAL PROCEDURE: -. SPECIALTY: Pulmonology. OBJECTIVE: Rare disease. BACKGROUND: Although pulmonary carcinoid tumors are generally considered to represent a low-grade malignancy, atypical carcinoids are more aggressive than typical carcinoids, metastasizing more commonly to both regional lymph nodes and distant sites. The treatment of choice for localized disease is surgery. In cases of advanced or metastatic disease, medical treatments, including chemotherapy, have not been proven to be very successful. Therefore, providing careful follow-up is extremely important. In general, tumor markers, such as the level of CYFLA21-1, are often useful for monitoring lung cancer. However, there are currently no sensitive tumor markers for carcinoid tumors. We herein report a rare case of an atypical carcinoid of the lung with the elevation of the serum ProGRP level. CASE REPORT: A 67-year-old female was referred to our hospital for an abnormal chest X-ray. CT revealed an 18×13 mm nodule in the right middle lobe with no significant mediastinal lymphadenopathy. The serum tumor marker, the ProGRP level, was significantly elevated (161 ng/ml). We performed a right middle lobectomy, because the pathological diagnosis of lung cancer was confirmed according to the results of a rapid frozen section biopsy of the lesion, although the pathological type could not be precisely determined by the frozen section alone. The final pathological diagnosis was atypical carcinoid. The level of ProGRP decreased (69 ng/ml) within 1 month after the surgery. CONCLUSIONS: The ProGRP level may be useful for monitoring carcinoid tumors, although no serum tumor markers are highly specific or sensitive for detecting recurrences and/or distant metastasis of pulmonary carcinoid tumors. In conclusion, ProGRP should be further evaluated as biomarker in a larger series of patients to determine whether it demonstrates any significant correlation with cancer recurrence.

Salvage operation for late recurrence after stereotactic body radiotherapy for lung cancer: two patients with no viable cancer cells.

We report two patients who underwent salvage lung resection for suspected local recurrence on computed tomography image findings after stereotactic body radiotherapy; however, the pathologic findings indicated no viable tumor cells. Distinguishing between posttreatment changes and tumor recurrence after stereotactic body radiotherapy on the image findings is difficult; therefore, the determination of surgical indications requires comprehensive evaluations.

Lung cancer mimicking lung abscess formation on CT images.

PATIENT: Male, 64 FINAL DIAGNOSIS: Lung pleomorphic carcinoma Symptoms: Cough • fever MEDICATION: - Clinical Procedure: - Specialty: Oncology. OBJECTIVE: Unusual clinical course. BACKGROUND: The diagnosis of lung cancer is often made based on computed tomography (CT) image findings if it cannot be confirmed on pathological examinations, such as bronchoscopy. However, the CT image findings of cancerous lesions are similar to those of abscesses.We herein report a case of lung cancer that resembled a lung abscess on CT. CASE REPORT: We herein describe the case of 64-year-old male who was diagnosed with lung cancer using surgery. In this case, it was quite difficult to distinguish between the lung cancer and a lung abscess on CT images, and a lung abscess was initially suspected due to symptoms, such as fever and coughing, contrast-enhanced CT image findings showing a ring-enhancing mass in the right upper lobe and the patient's laboratory test results. However, a pathological diagnosis of lung cancer was confirmed according to the results of a rapid frozen section biopsy of the lesion. CONCLUSIONS: This case suggests that physicians should not suspect both a lung abscesses and malignancy in cases involving masses presenting as ring-enhancing lesions on contrast-enhanced CT.

A case of synchronous double primary lung cancer presenting with pleomorphic carcinoma and adenocarcinoma.

BACKGROUND: Recently, synchronous multiple lung cancer (SMPLC) has sometimes been detected as a result of improved radiological imaging, although the occurrence of SPMLC is still rare. To the best of our knowledge, there have been no reported cases of with synchronous double primary lung cancer presenting with pleomorphic carcinoma and adenocarcinoma. We herein report such a case. CASE REPORT: A 64-year-old male was referred to our institution for an abnormal shadow in the apex of the left lung in April 2012. CT revealed 2 nodules that measured 15 mm in the left S(1+2b) and 20 mm in the left S3c. We suspected that the lesions were malignant, although the diagnosis could not be confirmed by transbronchial lung biopsy of the lesions. Therefore, we performed the left upper lobectomy. The results of the pathological examination of the nodule in S3c showed adenocarcinoma of pT1aN0M0, stage IA. The nodule in S(1+2b) was found to be pleomorphic carcinoma, pT1aN0M0, stage IA. In November 2012, the patient underwent an esophagogastroduodenoscopy because of anemia. The image findings showed a gastric ulcer on the greater curvature of his stomach. The pathological examination of the biopsy specimen from the ulcer revealed the metastatic cancer from pleomorphic carcinoma. In addition, abdominal CT revealed bilateral adrenal metastasis. Although the patient received chemotherapy, it was not effective. It was difficult to continue the chemotherapy because his performance status worsened. He died in May 2013. CONCLUSIONS: The present case was associated with a poor prognosis, even though the pathological stage of each tumor was stage IA. The prognosis of SMPLC may be associated with the histologic type, although the prognosis of SMPLC remains unclear due to its rarity.