Tropomyosin-related Kinase B Is Potentially a Biomarker of Prognosis and Therapeutic Target for Malignant Thymic Epithelial Tumors.

BACKGROUND/AIM: Thymic epithelial tumors (TETs) mainly consist of thymoma and thymic carcinoma. Complete surgical resection is vital for the successful management of these TETs, and adjuvant therapy such as systematic chemotherapy and/or radiotherapy plays important roles in the management of recurrent or metastasized disease. However, there is still a lack of a standard treatment after the failure of these adjuvant therapies. There is thus a need to develop molecular targeted therapies for advanced malignant TETs. In the present study, we evaluated the biological significance of brain-derived neurotrophic factor (BDNF)/tropomyosin receptor kinase B (TrkB) signaling for TETs. MATERIALS AND METHODS: The expression of TrkB in 48 formalin-fixed, paraffin-embedded TET specimens (43 thymoma and 5 thymic carcinoma) collected by surgical resection was evaluated immunohistochemically. A thymic carcinoma cell line was evaluated for the role of BDNF/TrkB signaling pathway in an in vitro assay. RESULTS: High TrkB expression was related to significantly poor prognosis in patients with TETs. In vitro experiments showed that BDNF/TrkB signaling was involved in the proliferation of Ty-82 cells, but not their invasion and migration. CONCLUSION: TrkB expression is a biomarker of the prognosis for TETs and the BDNF/TrkB signaling pathway is potentially a new therapeutic target for mTETs.

PTPN3 is a potential target for a new cancer immunotherapy that has a dual effect of T cell activation and direct cancer inhibition in lung neuroendocrine tumor.

In our previous study, we found that inhibition of protein tyrosine phosphatase non-receptor type 3 (PTPN3), which is expressed in lymphocytes, enhances lymphocyte activation, suggesting PTPN3 may act as an immune checkpoint molecule. However, PTPN3 is also expressed in various cancers, and the biological significance of PTPN3 in cancer cells is still not well understood, especially for lung neuroendocrine tumor (NET).Therefore, we analyzed the biological significance of PTPN3 in small cell lung cancer and examined the potential for PTPN3 inhibitory treatment as a cancer treatment approach in lung NET including small cell lung cancer (SCLC) and large cell neuroendocrine cancer (LCNEC). Experiments in a mouse xenograft model using allo lymphocytes showed that PTPN3 inhibition in SCLC cells enhanced the anti-tumor effect of PTPN3-suppressed activated lymphocytes. In addition, PTPN3 was associated with increased vascularization, decreased CD8/FOXP3 ratio and cellular immunosuppression in SCLC clinical specimens. Experiments in a mouse xenograft model using autocrine lymphocytes also showed that PTPN3 inhibition in LCNEC cells augmented the anti-tumor effect of PTPN3-suppressed activated lymphocytes. In vitro experiments showed that PTPN3 is involved in the induction of malignant traits such as proliferation, invasion and migration. Signaling from PTPN3 is mediated by MAPK and PI3K signals via tyrosine kinase phosphorylation through CACNA1G calcium channel. Our results show that PTPN3 suppression is associated with lymphocyte activation and cancer suppression in lung NET. These results suggest that PTPN3 suppression could be a new method of cancer treatment and a major step in the development of new cancer immunotherapies.

S100P regulates the collective invasion of pancreatic cancer cells into the lymphatic endothelial monolayer.

Lymph node metastasis is an independent prognostic factor in pancreatic cancer. However, the mechanisms of lymph node colonization are unknown. As a mechanism of lymphatic metastasis, it has been reported for other types of cancer that spheroids from tumor cells cause circular chemorepellent­induced defects (CCIDs) in lymphatic endothelial monolayers. In pancreatic cancer, such mechanisms of metastasis have not been elucidated. The present study evaluated the involvement of this new mechanism of metastasis in pancreatic cancer and investigated the associated factors. In human pancreatic cancer tissue, it was observed that clusters of cancer cells penetrated the wall of lymphatic ducts around the primary tumor. An in vitro co­culture system was then used to analyze the mechanisms of tumor cell­mediated disruption of lymphatic vessels. Time­lapse microscopic imaging revealed that spheroids from pancreatic cancer cells caused circular defects in lymphatic endothelial monolayers. CCID formation ability differed depending on the cell line. Neither aggregation of spheroids nor adhesion to lymphatic endothelial cells (LECs) exhibited a significant correlation with this phenomenon. The addition of supernatant from cultured cancer cells enhanced CCID formation. Microarray analysis revealed that the expression of S100 calcium binding protein P (S100P) was significantly increased when LECs were treated with supernatant from cultured cancer cells. Addition of a S100P antagonist significantly suppressed the migration of LECs and CCID formation. The present findings demonstrated that spheroids from pancreatic cancer cells caused circular defects in lymphatic endothelial monolayers. These CCIDs in pancreatic cancer were partly regulated by S100P, suggesting that S100P may be a promising target to inhibit lymph node metastasis.

Activities of daily living (ADL) of single elderly individuals using social assistive programs in a rural community.

Objectives　The proportion of elderly individuals living alone is increasing in Japan. Matsumoto city office provides social assistive programs such as home help, lunch delivery, life advice, and safety check telephone calls. The purpose of this study was to compare the level of ADL between the elderly using social assistive programs (the use group) and those who did not (the non-use group).Methods　We conducted a cross-sectional study at Shiga district of Matsumoto city in September 2014. A total of 128 elderly individuals participated in this study. Health volunteers asked these subjects to complete a questionnaire without assistance. Measurement items included lifestyle variables and social support networks. With respect to the frequency of use, we used questions that inquired about the use of the social assistive program. We included a set of instruments commonly used in the health assessment of elderly populations: functional capacity (Instrumental ADL, Intellectual Activity, Social Role), social support, nutrition (Mini Nutrition Assessment [MNA]) and depressive symptoms (Geriatric Depression Scale [GDS]).Results　The use group consisted of 24 elderly individuals participating in the social support program. The non-use group consisted of 89 elderly individuals living alone without programs. The mean age of those who completed the survey was 83.9±4.2 years for the use group and 82.3±4.3 years for the non-use group. Comparisons between the two groups did not show significant difference in terms of their intellectual activity, social role, emotional social support, and MNA or GDS scores. The use group was more likely to use the public transfer service and receive instrumental social support from children and relatives.Conclusions　By means of utilizing the public transfer service, and receiving family support, the elderly living alone who used social assistive programs could live independently. These findings suggest a need for improvement in the public transfer service and social network.

Availability, usage, and factors affecting usage of electrophysical agents by physical therapists: a regional cross-sectional survey.

[Purpose] The aim of this study was to investigate the availability, usage, and factors affecting usage of electrophysical agents by physical therapists in Nagano Prefecture, Japan. [Subjects and Methods] Questionnaires were sent to all 1,571 physical therapists working in 245 institutions within Nagano Prefecture. A total of 1,110 questionnaires were returned, out of which 1,099 (70%) questionnaires containing valid responses were analyzed. Frequencies and percentages were calculated for 22 modalities with regards to availability, usage, rate of usage, and confidence level in usage. Factors affecting usage and the relationship between rate of usage and confidence level (Spearman's rho) were also determined. [Results] The top three responses for the various outcome measures were as follows: (1) hot packs (88%), low frequency stimulators (76%), and ultrasound (68%) for availability; (2) hot packs (72%), ultrasound (61%), and cold packs (59%) for usage; (3) hot packs (75%), cold spray (49%), and ultrasound (44%) for confidence in usage; and (4) equipment availability (80%), past experience (79%), and research evidence (78%) for factors affecting usage. There was a significant positive relationship between confidence and usage for all modalities, except for ultraviolet radiation, iontophoresis, and magnetic field. [Conclusion] Usage was strongly correlated with confidence, with the top three used modalities also being the ones with the highest confidence in usage.

[A case of squamous cell carcinoma of the nipple skin successfully treated with S-1 alone].

Squamous cell carcinoma of the breast is uncommon, but that of the nipple skin is rarer. The effect of chemotherapy in these cases is yet to reach consensus. We report a rare case in which primary squamous cell carcinoma of the nipple skin was successfully treated with S-1 alone. A 64-year-old woman was admitted to our hospital because of a granulomatous tumor mass over the right nipple, which she was aware of for 10 years; the tumor showed a rapid increase in growth before admission. The tumor was approximately 4 cm at the first visit, and was diagnosed as squamous cell carcinoma by incisional biopsy. We administered preoperative systemic chemotherapy owing to the presence of metastasis in an axillary lymph node. After 2 courses of chemotherapy with oralS -1 at 100mg/day for 28 days followed by a 14-day resting period, the primary tumor and metastatic lymph node showed a remarkable reduction in size. The patient subsequently underwent a radical operation and is currently healthy without any recurrence.

Interaction between NANOS2 and the CCR4-NOT deadenylation complex is essential for male germ cell development in mouse.

Nanos is one of the evolutionarily conserved proteins implicated in germ cell development and we have previously shown that it interacts with the CCR4-NOT deadenylation complex leading to the suppression of specific RNAs. However, the molecular mechanism and physiological significance of this interaction have remained elusive. In our present study, we identify CNOT1, a component of the CCR4-NOT deadenylation complex, as a direct factor mediating the interaction with NANOS2. We find that the first 10 amino acids (AAs) of NANOS2 are required for this binding. We further observe that a NANOS2 mutant lacking these first 10 AAs (NANOS2-ΔN10) fails to rescue defects in the Nanos2-null mouse. Our current data thus indicate that the interaction with the CCR4-NOT deadenylation complex is essential for NANOS2 function. In addition, we further demonstrate that NANOS2-ΔN10 can associate with specific mRNAs as well as wild-type NANOS2, suggesting the existence of other NANOS2-associated factor(s) that determine the specificity of RNA-binding independently of the CCR4-NOT deadenylation complex.

Analysis of pelvic movement in the elderly during walking using a posture monitoring system equipped with a triaxial accelerometer and a gyroscope.

The incidence of falls in the elderly is increasing with the aging of society and is becoming a major public health issue. From the viewpoint of prevention of falls, it is important to evaluate the stability of the gait in the elderly people. The pelvic movement, which is a critical factor for walking stability, was analyzed using a posture monitoring system equipped with a triaxial accelerometer and a gyroscope. The subjects were 95 elderly people over 60 years of age. The criteria for instability were open-eye standing on one leg for 15s or less, and 11s or more on 3m timed up and go test. Forty subjects who did not meet both of these criteria comprised the stable group, and the remaining 55 subjects comprised the unstable group. Pelvic movement during walking was compared between the two groups. The angle, angular velocity, and acceleration were analyzed based on the wave shape derived from the device worn around the second sacral. The results indicated that pelvic movement was lower in all three directions in the unstable group compared to the stable group, and the changes in the pelvic movement during walking in unstable elderly people were also reduced. This report is the first to evaluate pelvic movement by both a triaxial accelerometer and a triaxial gyroscope simultaneously. The characteristics of pelvic movement during walking can be applied in screening to identify elderly people with instability, which is the main risk factor associated with falls.

VIO soft-coagulation system for major pulmonary resections: results in 68 patients with primary lung cancer.

PURPOSE: The VIO soft-coagulation system is a new device for tissue coagulation in which the output voltage is automatically regulated. We evaluated the safety and efficacy of this system for major pulmonary resection of lung cancer. METHODS: We used bipolar VIO soft-coagulation for major pulmonary resection in 68 patients with primary lung cancer using the following three surgical procedures: (1) coagulation of the tissues around lobar vessels and bronchus; (2) fissure dissection; (3) lymph node dissection. The dissection in those steps was performed with both bipolar VIO soft-coagulation forceps and scissors. Results were analyzed with special reference to intraoperative complications, blood loss, and postoperative morbidity. The data in this study (VIO group) were compared with those of 78 lung cancer patients who underwent surgery using conventional electrocautery (CE group). RESULTS: The operations in this series included 60 lobectomies, 1 bilobectomy, 1 pneumonectomy, and 6 segmentectomies. With the VIO soft-coagulation system, coagulation of tissues around the lobar pulmonary vessels without injury to the pulmonary vessels or bronchus was effective. The mean blood loss in the VIO group was 90.6 g (range 5-595 g), which was significantly lower than that in the CE group (mean 141.0 g, range 5-700 g) (P = 0.014). Postoperative morbidity rates in the VIO and the CE group were 11.7% and 17.9%, respectively. CONCLUSION: The VIO soft-coagulation system is safe and feasible for major pulmonary resections in patients with lung cancer.

Surgical treatment of clinical N1 non-small cell lung cancer: ongoing controversy over diagnosis and prognosis.

PURPOSE: The preoperative assessment of nodal status in lung cancer is complicated and problematic for physicians and surgeons. Although many patients with clinical N1 (cN1) non-small cell lung cancer (NSCLC) are candidates for surgical treatment, these patients represent a heterogeneous subgroup with unpredictable survival. We conducted this study to evaluate the surgical results of cN1 disease and to attempt to clarify the delicate issues surrounding its diagnosis and prognosis. METHODS: The subjects of this study were 187 consecutive patients with cN1 adenocarcinoma or squamous cell carcinoma of the lung, who underwent complete resection without induction therapy. RESULTS: Only 25% of the adenocarcinomas and 54% of the squamous cell carcinomas were correctly diagnosed as N1 disease preoperatively. Multiple logistic regression analyses revealed that adenocarcinoma (P = 0.0141) was a significant predictor of pN2. Multivariate analyses revealed that nodal metastasis (P < 0.0001), large tumor size (P = 0.0079), and high serum carcinoembryonic antigen value (P = 0.0096) were significantly poor prognostic factors in cN1 patients. CONCLUSIONS: It is difficult to diagnose nodal status in patients with cN1 disease, which requires various surgical procedures, including plasty, possibly with adjuvant therapy in a defined high-risk subgroup.

SPARC mRNA expression as a prognostic marker for pancreatic adenocarcinoma patients.

BACKGROUND: Secreted protein acidic and rich in cysteine (SPARC) is a protein which governs diverse cellular functions and matrix remodeling systems. It was reported that SPARC expression of peritumoral fibroblasts was correlated with poor prognosis in resectable pancreatic cancer patients in immunohistochemical analysis. However, the significance of SPARC mRNA expression in pancreatic adenocarcinoma remains unclear. MATERIALS AND METHODS: SPARC mRNA expression was evaluated in 104 formalin-fixed paraffin-embedded tissue samples of pancreatic adenocarcinoma patients with macro-dissection technique. RESULTS: High SPARC mRNA expression was associated with a poorer prognosis than low SPARC. In univariate Cox's proportion hazard model, depth of tumor invasion, lymph node metastasis, histopathological tumor grade, lymphatic invasion, vascular invasion, surgical margin, no chemotherapy and high SPARC expression were significant prognostic factors. In multivariate analysis, histopathological tumor grade, surgical margin, no chemotherapy and high SPARC expression were independent significant prognostic factors. CONCLUSION: SPARC mRNA expression is a prognostic marker for pancreatic adenocarcinoma patients.

Beneficial effect of combination therapy comprising angiotensin II receptor blocker plus calcium channel blocker on plasma adiponectin levels.

The study aim was to examine the effect of combination therapy comprising angiotensin receptor blocker plus calcium antagonist on post-treatment plasma adiponectin levels compared to pretreatment levels. There was a significant gender difference in the relationship between preadiponectin level and age. In the search for contributing factors for treatment-based changes in adiponectin levels, these effects of gender and age were considered in statistical analysis. The adiponectin level in the combination therapy group was further increased compared to that in each of the monotherapy groups, despite there being no significant difference in antihypertensive effect, indicating that the combined medication provided an effect beyond that of lowering blood pressure.

High efficacy of disopyramide in the management of ventricular fibrillation storms in a patient with Brugada syndrome.

The patient was a 57-year-old man with Brugada syndrome, who had been implanted with a implantable cardioverter defibrillator (ICD). The frequency of ventricular fibrillation (VF) started to increase about 4 years after ICD implantation, occurring, at worst, six times in one night. Immediately after starting oral administration of disopyramide, VF stopped occurring. He then discontinued taking disopyramide, but immediately after the discontinuation VF started occurring again, so he restarted taking disopyramide. Thereafter, VF completely stopped occurring. Findings observed in our case suggest that disopyramide could be added in our arsenal of medications for treating arrhythmic storms in patient with Brugada syndrome.

Additive antihypertensive and antihypertrophic effects of long-acting Ca blockers in uncontrolled hypertensive patients with angiotensin-receptor blocker based treatment.

The aim of the present study was to examine the antihypertensive and antihypertrophic effects of combined treatment with a long-acting calcium antagonist on top of an angiotensin II receptor blocker (ARB) in uncontrolled hypertensive patients. Patients with essential hypertension and a blood pressure > 140/90 mmHg on ARB monotherapy (losartan 50 mg/day or candesartan 8 mg/day) were randomly assigned to a nifedipine controlled release (CR) group (n = 15) or amlodipine group (n = 11). A significant additional antihypertensive effect was noted from 1 month with nifedipine and 2 months with amlodipine. The average daily dose was 25 mg for nifedipine and 5 mg for amlodipine. The cardiothoracic ratio was significantly reduced in both groups after 3 months. Left ventricular wall thickness and left ventricular mass index also decreased. Metabolic parameters, hepatic function, and renal function did not change significantly. Additional treatment with a long-acting calcium antagonist achieved further blood pressure reduction as well as an antihypertrophic effect in the uncontrolled patients with prior ARB monotherapy.

Tumor-stromal interactions with direct cell contacts enhance proliferation of human pancreatic carcinoma cells.

Pancreatic ductal adenocarcinoma is often characterized by an abundant desmoplastic stroma that is partially induced by activated pancreatic stellate cells (PSCs). Indirect co-culture has often been used to investigate the effects of cancer-stromal interactions on the proliferation of cancer cells, but the effects of cell-cell adhesion and juxtacrine signaling between cancer and stromal cells cannot be evaluated using this method. This study aimed to establish a simplified direct co-culture system that could be used to quantify populations of cancer cells in co-culture with PSCs, and to evaluate the effects of direct cell contact on the proliferation of cancer cells. We established three green fluorescent protein (GFP)-expressing pancreatic cancer cell lines and were able to quantify them with high reliability and reproducibility, even when co-cultured directly with PSCs, using a color plate reader. We assessed the differential effects of direct and indirect co-culture with PSCs on the proliferation of cancer cells, and found that the proliferation of GFP-expressing pancreatic cancer cell lines was dramatically enhanced by direct co-culture with PSCs, compared with the indirect co-culture system. We also found that direct co-culture of cancer cells and PSCs activated the Notch signaling pathway in both cell types. Direct cell contact between cancer cells and PSCs plays an important role in the control of cancer cell proliferation, and is essential to the understanding of tumor-stromal interactions.

Takotsubo cardiomyopathy complicated by high-grade atrioventricular block: A report of two cases.

The present report describes two female patients aged 39 and 57 years who experienced loss of consciousness and chest pain due to high-grade atrioventricular block. Both patients demonstrated noncontraction centred on the cardiac apex and excessive contraction at the cardiac base on cardiac ultrasonography and left ventriculography, but neither of them demonstrated any significant stenotic lesions on coronary angiography. Furthermore, neither patient showed elevated serum biomarkers of cardiac injury or serum viral antibodies. In a repeat left ventriculogram two weeks later, the left ventricular wall motion disorder had improved in both patients. Based on these findings, the patients were diagnosed with takotsubo cardiomyopathy. Because the high-grade atrioventricular conduction disorder did not improve in spite of the improvement of left ventricular wall motion disorder, permanent pacemaker implantation was performed. It is extremely rare for takotsubo cardiomyopathy to be complicated by high-grade atrioventricular block. In the present study, both patients had takotsubo cardiomyopathy complicated by high-grade atrioventricular block and eventually underwent permanent pacemaker implantation.

Ventricular pacing thresholds following high-energy implantable cardioverter defibrillator shocks in integrated bipolar defibrillation systems.

BACKGROUND: Increased ventricular pacing thresholds have been observed following monophasic implantable cardioverter defibrillator (ICD) shocks. AIM: To examine changes following high-energy biphasic shocks delivered by integrated bipolar ICD systems. METHOD: Ten episodes of ventricular fibrillation (VF) were induced at 10 min intervals in nine pigs with integrated ICD systems. After 10 s of each episode of VF, a 40 J biphasic shock was delivered, which successfully terminated VF (a total of 10 shocks). The bipolar pacing threshold at the right ventricular apex was measured before each shock and at 1 min intervals after each shock. RESULTS: The mean pacing threshold was 0.029+/-0.059 muJ before the first shock and gradually increased to 0.14+/-0.10 muJ after the 10th shock. CONCLUSION: It may be necessary to pace at a high-voltage output following biphasic shocks delivered by integrated bipolar ICD systems.

Chemotherapeutic agents potentiate adenoviral gene therapy for pancreatic cancer.

Adenovirus-mediated gene therapy combined with chemotherapeutic agents is expected to represent a new approach for treating pancreatic cancer. However, there have been no reports of definitive effects of chemotherapeutic agents on adenovirus-mediated gene therapies. In the present study, we investigated the effects of chemotherapeutic agents on the transduction efficiency of an adenovirus-based gene therapy. Adenovirus (Ad-NK4) expressing NK4, which acts as a hepatocyte growth factor antagonist, was used as a representative gene therapy. Pancreatic cancer cells infected with Ad-NK4 were treated with chemotherapeutic agents (5-fluorouracil [5FU], cisplatin or etoposide), and the NK4 levels in their culture media were measured. To examine the effects of chemotherapeutic agents in vivo, Ad-NK4 was administered to subcutaneous tumors in mice after treatment with the agents, and the tumor NK4 levels were measured. The NK4 levels in culture media from cells treated with 5FU, cisplatin and etoposide were 5.2-fold (P = 0.026), 6-fold (P < 0.001) and 4.3-fold (P < 0.001) higher than those of untreated cells, respectively. The chemotherapeutic agents also increased Ad-NK4 uptake. The NK4 levels in tumors treated with 5FU, cisplatin and etoposide were 5.4-fold (P = 0.006), 11.8-fold (P < 0.001) and 4.9-fold (P = 0.017) higher than those in untreated tumors, respectively. The present findings suggest that chemotherapeutic agents significantly improve the efficiency of adenovirus-mediated gene transfer in pancreatic cancer. Furthermore, they will contribute to decreases in the adenovirus doses required for gene transfer, thereby controlling the side-effects of adenovirus infection in normal tissues.

Mercury-free preparation and selective reactions of propargyl (and propargylic) Grignard reagents.

ZnBr2 was found to catalyze formation of propargyl and propargylic Grignard reagents, and thus put an end to the standard method using a mercury catalyst. The Grignard reagents were submitted to addition reaction with carbonyl compounds and allylation with the cyclic monoacetate to afford the propargyl-type products selectively. Furthermore, the product from the monoacetate was transformed to an acetylene analogue of 2-(5,6-epoxyisoprostane A2)phosphorylcholines.