Upregulation and stabilization of senescence marker protein-30 by epigallocatechin gallate against tert-butyl hydroperoxide-induced liver injury in vitro and in vivo.

Senescence marker protein-30 (SMP30), a novel ageing marker, suppresses oxidative stress in the liver. However, studies on phytochemical-mediated regulation of SMP30 expression are lacking. Here, we showed that epigallocatechin gallate (EGCg), a polyphenol abundant in green tea, positively regulates SMP30 expression in the rat hepatoma-derived Fao cells. EGCg maintained SMP30 expression even in the presence of cycloheximide, a protein synthesis inhibitor. Furthermore, treatment of cells with tert-butyl hydroperoxide (tert-BHP), an oxidative promoter, decreased SMP30 expression and ERK1/2 phosphorylation, while EGCg treatment inhibited these effects. Male mice (7-week-old) were divided into 4 groups-Control (saline), tert-BHP (1.5 mmol/kg tert-BHP), EGCg + tert-BHP (30 mg/kg/day of EGCg and 1.5 mmol/kg tert-BHP), and EGCg (30 mg/kg/day). After oral EGCg administration for 6 consecutive days, EGCg + tert-BHP group mice were administered tert-BHP. The tert-BHP-administered mice showed decreased SMP30 expression in the liver and increased aspartate aminotransferase and alanine transaminase (hepatic injury marker enzymes) activities; however, EGCg treatment attenuated these changes. Thus, EGCg-induced SMP30 upregulation may alleviate tert-BHP-induced liver injury. The findings of this study offer new perspectives of the anti-ageing properties of EGCg.

Experiences and hidden needs of older patients, their families and their physicians in palliative chemotherapy decision-making: a qualitative study.

OBJECTIVE: This study aimed to clarify the experiences and hidden needs of older patients with advanced cancer, their families and their physicians in palliative chemotherapy decision-making. MATERIALS AND METHODS: We conducted in-depth qualitative individual interviews with content analysis. Patients who were diagnosed as having advanced cancer, were aged ≥70 years (n = 15, median [range] = 77 [70-82] years) and had volunteered to receive palliative chemotherapy within the past 6 months were enrolled. Their families and physicians were also interviewed. RESULTS: The following four themes were identified: (i) physician's awareness of paternalism; (ii) readiness for communication of serious news; (iii) spiritual care need assessment and (iv) support as a team. The patients and families expected physicians to demonstrate paternalism in their decision-making because they were unconfident about their self-determination capability. Although the physicians were aware of this expectation, they encountered difficulties in recommending treatment and communicating with older patients. The patients had spiritual pain since the time of diagnosis. Psychological issues were rarely discussed during decision-making and treatment, triggering feelings of isolation in the patients and their families. CONCLUSION: Older patients and their families expected a paternalistic approach by the physicians for palliative chemotherapy decision-making. The physicians found it difficult to offer treatment options because of older patient diversity and limitations in evidence-based strategies. Therefore multidisciplinary approaches and evidence-based decision support aids are warranted. Because older patients and their families often have unexpressed psychological burdens including unmet spiritual needs, medical professionals should provide psychological care from the time of diagnosis.

Down-regulation of senescence marker protein 30 by iron-specific chelator deferoxamine drives cell senescence.

To our knowledge, this is the first study to report down-regulation of senescence marker protein 30 (SMP30) by iron-specific chelator deferoxamine (DFO) on FAO cell senescence, using a DNA microarray. Furthermore, DFO treatment increased senescence marker ß-galactosidase activity, whereas this activity was attenuated by overexpression of SMP30. Our data suggested that down-regulation of SMP30 drives cell senescence in iron-chelated condition.

Sulforaphane inhibits osteoclast differentiation by suppressing the cell-cell fusion molecules DC-STAMP and OC-STAMP.

Sulforaphane (SFN), a kind of isothiocyanate, is derived from broccoli sprouts. It has anti-tumor, anti-inflammatory, and anti-oxidation activity. The molecular function of SFN in the inhibition of osteoclast differentiation is not well-documented. In this study, we assessed the effect of SFN on osteoclast differentiation in vitro. SFN inhibited osteoclast differentiation in both bone marrow cells and RAW264.7 cells. Key molecules involved in the inhibitory effects of SFN on osteoclast differentiation were determined using a microarray analysis, which showed that SFN inhibits osteoclast-associated genes, such as osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells cytoplasmic-1, tartrate-resistant acid phosphatase, and cathepsin K. Moreover, the mRNA expression levels of the cell-cell fusion molecules dendritic cell specific transmembrane protein (DC-STAMP) and osteoclast stimulatory transmembrane protein (OC-STAMP) were strongly suppressed in cells treated with SFN. Furthermore, SFN increased the phosphorylation of signal transducer and activator of transcription 1 (STAT1), a regulator of macrophage and osteoclast cell fusion. Thus, our data suggested that SFN significantly inhibits the cell-cell fusion molecules DC-STAMP and OC-STAMP by inducing the phosphorylation of STAT1 (Tyr701), which might be regulated by interactions with OSCAR.

Sulforaphene attenuates multinucleation of pre-osteoclasts by suppressing expression of cell-cell fusion-associated genes DC-STAMP, OC-STAMP, and Atp6v0d2.

We assessed the effect of sulforaphene (SFE) on osteoclast differentiation. SFE significantly decreased the number of RANKL-induced tartrate-resistant acid phosphatase-positive cells and suppressed pre-osteoclast multinucleation. Furthermore, SFE downregulated mRNA expression of DC-STAMP, OC-STAMP, and Atp6v0d2, which encode cell-cell fusion molecules. Our data suggest that SFE attenuates pre-osteoclast multinucleation via suppression of cell-cell fusion.

Extracts of black and brown rice powders improve hepatic lipid accumulation via the activation of PPARα in obese and diabetic model mice.

Rice powder extract (RPE) from black and brown rice (Oryza sativa L. indica) improves hepatic lipid accumulation in obese and diabetic model mice via peroxisomal fatty acid oxidation. RPE showed PPARα agonistic activity which did not differ between black and brown RPE despite a higher anthocyanin content in black RPE.

Iron deficiency negatively regulates protein methylation via the downregulation of protein arginine methyltransferase.

Iron is an essential trace metal for all biological processes and plays a role in almost every aspect of body growth. Previously, we found that iron-depletion downregulated the expression of proteins, arginine methyltransferase-1 and 3 (PRMT1 and PRMT3), by an iron-specific chelator, deferoxamine (DFO), in rat liver FAO cell line using DNA microarray analysis (unpublished data). However, regulatory mechanisms underlying the association between iron deficiency and PRMT expression are unclear in vitro and in vivo. In the present study, we revealed that the treatment of cells with two iron-specific chelators, DFO and deferasirox (DFX), downregulated the gene and protein expression of PRMT1 and 3 as compared with the untreated cells. Subsequently, DFO and DFX treatments decreased protein methylation. Importantly, these effects were attenuated by a holo-transferrin treatment. Furthermore, weanling Wistar-strain rats were fed a control diet or an iron-deficient diet for 4 weeks. Dietary iron deficiency was found to decrease the concentration of hemoglobin and liver iron while increasing the heart weight. PRMT and protein methylation levels were also significantly reduced in the iron-deficient group as compared to the control group. To our knowledge, this is the first study to demonstrate that PRMT levels and protein methylation are reduced in iron-deficient models, in vitro and in vivo.

Severe iron deficiency decreases both bone formation and bone resorption in rats.

The purpose of this study was to clarify the manner in which dietary iron deficiency decreased bone mineral density (BMD) in rats. Eighteen 3-wk-old male Wistar rats were divided into 3 groups of 6 rats each. The rats in 2 of the 3 groups had free access to a control diet (C group) or an iron-deficient diet (ID group) for 4 wk. The rats in the third group (PF group) were pair-fed the control diet to the mean intake of the ID group. Compared with the C and PF groups, hematocrit and hemoglobin concentrations were significantly reduced and bone mineral content and BMD of the femur were significantly lower in the ID group. Bone histomorphometric parameters showed that the bone formation rate and osteoclast surface in the lumbar vertebra were significantly reduced in the ID group compared with the C and PF groups. Furthermore, dietary iron deficiency decreased serum 1,25-dihydroxycholecalciferol, insulin-like growth factor-I, and osteocalcin concentrations and urinary excretion of deoxypyridinoline. These results suggest that severe iron deficiency decreases not only bone formation but also bone resorption.

Iron deficiency induces autophagy and activates Nrf2 signal through modulating p62/SQSTM.

Iron is an essential trace metal in almost all organisms and plays an important role in the redox system. We previously reported that iron deficiency activated autophagy and nuclear factor erythroid 2-related factor 2 (Nrf2) signaling for oxidative stress. However, regulatory mechanisms underlying the association between autophagy and Nrf2 signaling are unclear. In this study, we found that treatment of cells with an iron-specific chelator deferoxamine (DFO) increased reactive oxidative species (ROS) production by elevating the expression of p47phox and p67phox compared with that in untreated cells. The DFO treatment also induced protein aggregation and formed aggresome, which is a cellular response to misfolded protein. In addition, DFO treatment upregulated the expression of the autophagic gene p62/SQSTM1, which in turn activated intracellular proteolysis during autophagy. DFO treatment phosphorylated p62/SQSTM1 (Thr351) to activate Nrf2. However, silencing of p62/SQSTM1 followed by DFO treatment attenuated Nrf2 activation and resulted in the accumulation of carboxyl proteins compared with DFO treatment alone. These results indicated that iron deficiency activates Nrf2 signaling by modulating p62/SQSTM1 during autophagy.

Endobronchial ultrasound-guided transbronchial needle aspiration is useful as an initial procedure for the diagnosis of lymphoma.

BACKGROUND: The usefulness of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for evaluating hilar, mediastinal and central parenchymal lesions has been well established. However, its utility for diagnosing lymphoma is controversial. The aim of this study was to evaluate the diagnostic utility of EBUS-TBNA for the definitive diagnosis of de novo lymphoma with subtype classification. METHODS: Patients with lymphoma who underwent EBUS-TBNA for diagnostic purposes at a single institution between March 2004 and May 2013 were retrospectively reviewed. RESULTS: Of the 971 patients who underwent EBUS-TBNA during the study period, 19 patients, who did not have a previous history of lymphoma, had a final diagnosis of lymphoma. EBUS-TBNA provided a diagnosis accompanied with subtype classification in 6 patients (32%), a suspicious but not definitive classification in 10 patients (53%), and a negative classification in 3 patients (16%). Immunohistochemical staining for definitive diagnosis was performed in 15 of 16 patients (94%), with suspicious results from routine hematoxylin and eosin staining. No procedure-related complications occurred. CONCLUSIONS: EBUS-TBNA is a useful initial diagnostic procedure, aiding decisions for the management of patients with suspected lymphoma, even though the sensitivity of EBUS-TBNA for diagnosing lymphoma with subtype classification was lower than previously reported.

Safety and Complications of Medical Thoracoscopy.

Objectives. To highlight the possible complications of medical thoracoscopy (MT) and how to avoid them. Methods. A retrospective and prospective analysis of 127 patients undergoing MT in Nagoya Medical Center (NMC) and Toyota Kosei Hospital. The data about complications was obtained from the patients, notes on the computer system, and radiographs. Results. The median age was 71.0 (range, 33.0-92.0) years and 101 (79.5%) were males. The median time with chest drain after procedure was 7.0 (range, 0.0-47.0) days and cases with talc poudrage were 30 (23.6%). Malignant histology was reported in 69 (54.3%), including primary lung cancer in 35 (27.5), mesothelioma in 18 (14.2), and metastasis in 16 (12.6). 58 (45.7%) revealed benign pleural diseases and TB was diagnosed in 15 (11.8%). 21 (16.5%) patients suffered from complications including lung laceration in 3 (2.4%), fever in 5 (3.9%) (due to hospital acquired infection (HAI) in 2, talc poudrage in 2, and malignancy in 1), HAI in 2 (1.6%), prolonged air-leak in 14 (11.0%), and subcutaneous emphysema in 1 (0.8%). Conclusions. MT is generally a safe procedure. Lung laceration is the most serious complication and should be managed well. HAI is of low risk and can be controlled by medical treatment.

Rigid bronchoscopic intervention for endobronchial metastasis of renal cell carcinoma.

BACKGROUND: Renal cell carcinoma is one of the major endobronchial metastases, and it occasionally causes life-threatening airway obstruction. Rigid bronchoscopy is useful as a palliative intervention; however, its utility for metastatic renal cell carcinoma has not been elucidated. The purpose of this study was to evaluate the safety and efficacy of rigid bronchoscopic treatment for endobronchial metastasis of renal cell carcinoma. METHODS: Among 550 patients who underwent rigid bronchoscopic intervention at a single center from January 2005 to June 2015, 9 with metastatic renal cell carcinoma were retrospectively reviewed. Procedures were performed with rigid and flexible bronchoscopes under general anesthesia. RESULTS: In total, 20 procedures were performed on 9 patients who underwent stent implantation. Bleeding was observed in 12 (60%) procedures while severe hypoxia was observed in 2 (10%). The required amount of supplemental oxygen successfully decreased after all the 10 procedures (100%) in patients who previously needed it. Median survival after the first procedure was of 260 days (range, 63-913 days). CONCLUSIONS: Rigid bronchoscopic intervention for endobronchial metastasis of renal cell carcinoma is feasible with safety and effectiveness for palliation of airway obstruction, if one prepares sufficiently for massive intraoperative bleeding.

Transbronchial vs transesophageal needle aspiration using an ultrasound bronchoscope for the diagnosis of mediastinal lesions: a randomized study.

BACKGROUND: The purpose of this study was to compare the tolerance, efficacy, and safety of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) with transesophageal endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) with an endobronchial ultrasound scope for the first pathologic diagnosis of lesions accessible by both procedures. METHODS: Patients who had lesions accessible by both EBUS-TBNA and EUS-FNA were enrolled and were randomized to undergo either procedure. Patients quantified tolerance, and operators charted the quality of examination using a 100-mm visual analog scale (VAS). RESULTS: A specific diagnosis was made in 50 of 55 patients (91%) in the EBUS-TBNA group and in 48 of 55 patients (87%) in the EUS-FNA group (P = .76). Compared with EBUS-TBNA, EUS-FNA was associated with a shorter duration of procedure (median, 15.3 min vs 11.3 min; P < .001), lower doses of IV midazolam (mean, 4.4 mg vs 4 mg; P = .02) and intraairway lidocaine (mean, 303 mg vs 189 mg; P < .001), less frequent oxygen desaturations (23 of 55 vs two of 55, P < .001), and higher operator satisfaction (P < .001). There was no significant difference in patient tolerance according to the patients' VAS. Lymph node infection occurred in one patient in the EBUS-TBNA group and in two patients in the EUS-FNA group. CONCLUSIONS: Both EBUS-TBNA and EUS-FNA provide high accuracy with good tolerance, although the occurrence of infectious complications should be monitored carefully. EUS-FNA has the advantage of comparable tolerance with fewer doses of anesthetics and sedatives, a shorter procedure time, and fewer oxygen desaturations during the procedure. TRIAL REGISTRY: UMIN Clinical Trials Registry; No.: UMIN000005757; URL: http://www.umin.ac.jp/ctr/.

Effects of high phosphorus diet on bone metabolism-related gene expression in young and aged mice.

In this study, the effects of high phosphorus (P) diet on bone metabolism-related gene expression were investigated in young and aged mice. Twelve- and 80-week-old ddY male mice were divided into two groups, respectively, and fed a control diet containing 0.3% P or a high P diet containing 1.2% P. After 4 weeks of treatment, serum parathyroid hormone (PTH) concentration was significantly higher in the high P groups than in the control groups in both young and aged mice and was significantly higher in aged mice than in young mice fed the high P diet. High P diet significantly increased receptor activator of NF-κB ligand (RANKL) mRNA in the femur of both young and aged mice and significantly increased the RANKL/osteoprotegerin (OPG) mRNA ratio only in aged mice. High P diet significantly increased mRNA expression of transient receptor potential vanilloid type 6, calbindin-D9k, and plasma membrane Ca(2+)-ATPase 1b in the duodenum of both young and aged mice. These results suggest that high P diet increased RANKL mRNA expression in the femur and calcium absorption-related gene expression in the duodenum regardless of age. Furthermore, the high P diet-induced increase in PTH secretion might increase the RANKL/OPG mRNA ratio in aged mice.

Endobronchial ultrasound-guided transbronchial needle aspiration in the diagnosis of small cell lung cancer.

BACKGROUND: Massive lymphadenopathy and direct mediastinal invasion are well-recognized phenomena in patients with small cell lung cancer (SCLC). The aim of this study was to assess the utility of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of SCLC. METHODS: We retrospectively reviewed the records of 780 patients who underwent EBUS-TBNA at our institution from March 2004 to June 2012. Of these, 101 had a final diagnosis of SCLC. Excluding 3 patients with known SCLC who underwent EBUS-TBNA for staging purposes and including 2 patients who underwent EBUS-TBNA twice for the diagnosis of recurrence after achieving complete response by chemoradiation therapy during the study period, a total of 100 EBUS-TBNA procedures in 98 patients were analyzed. RESULTS: Other diagnostic tests prior to the initial EBUS-TBNA had failed to yield a diagnosis in 41 patients. The overall diagnostic yield of EBUS-TBNA for SCLC was 97% (97 of 100). Rapid on-site cytologic evaluation (ROSE) was performed at the operator's discretion in 77 procedures. ROSE did not have any impact on diagnostic yield (99% with ROSE vs. 90% without ROSE, p=0.1), but the use of ROSE was associated with fewer lesions (mean 1.1 with ROSE vs. 1.6 without ROSE, p<0.01) or aspirates (mean 2.3 with ROSE vs. 4.0 without ROSE, p<0.01). CONCLUSIONS: EBUS-TBNA provided a high diagnostic yield in SCLC with or without ROSE. EBUS-TBNA can be recommended for patients suspected to have SCLC, even if other diagnostic tests have failed.

Dietary iron deficiency decreases serum osteocalcin concentration and bone mineral density in rats.

We investigated the effects of dietary iron deficiency on bone metabolism by measuring markers of bone turnover in rats. Twelve 3-week-old male Wistar-strain rats were fed a control diet or an iron-deficient diet for 4 weeks. Dietary iron deficiency decreased hemoglobin concentration and increased heart weight. Serum osteocalcin concentration, bone mineral content, bone mineral density, and mechanical strength of the femur were significantly lower in the iron-deficient group than in the control group. These results suggested that dietary iron deficiency affected bone, which might have been due to a decrease in bone formation in rats.