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Severe hypertriglyceridemia is a pathological condition caused by genetic factors alone or in combination with environmental factors, sometimes leading to acute pancreatitis (AP). In this study, exome sequencing and biochemical analyses were performed in 4 patients with hypertriglyceridemia complicated by obesity or diabetes with a history of AP or decreased post-heparin LPL mass. In a patient with a history of AP, SNP rs199953320 resulting in LMF1 nonsense mutation and APOE rs7412 causing apolipoprotein E2 were both found in heterozygous form. Three patients were homozygous for APOA5 rs2075291, and one was heterozygous. ELISA and Western blot analysis of the serum revealed the existence of apolipoprotein A-V in the lipoprotein-free fraction regardless of the presence or absence of rs2075291; furthermore, the molecular weight of apolipoprotein A-V was different depending on the class of lipoprotein or lipoprotein-free fraction. Lipidomics analysis showed increased serum levels of sphingomyelin and many classes of glycerophospholipid; however, when individual patients were compared, the degree of increase in each class of phospholipid among cases did not coincide with the increases seen in total cholesterol and triglycerides. Moreover, phosphatidylcholine, lysophosphatidylinositol, and sphingomyelin levels tended to be higher in patients who experienced AP than those who did not, suggesting that these phospholipids may contribute to the onset of AP. In summary, this study revealed a new disease-causing gene mutation in LMF1, confirmed an association between overlapping of multiple gene mutations and severe hypertriglyceridemia, and suggested that some classes of phospholipid may be involved in the pathogenesis of AP.

Hydrogen gas is attractive as a clean fuel source if it can be produced efficiently without relying on fossil fuels. Biohydrogen production using photosynthetic bacteria may enable environmentally friendly hydrogen production but is currently limited by factors such as low oxygen tolerance. In this study, we isolate a new strain of bacteria that can produce hydrogen under aerial-phase conditions compared with those under liquid-phase conditions in a nitrogen gas or an argon gas atmosphere. Bacterial strains were cultured from scrapings taken from a steel signboard. Investigation of the hydrogen production of the strains under aerial- and liquid-phase conditions and subsequent DNA sequencing led to identification of the bacterium Cereibacter sp. KGU-NF001. Aerial-phase conditions were achieved by filter membranes with the bacterial strains and placing the membranes on medium-soaked cotton wool. The gas atmosphere affected the behavior of the isolated bacterial strains under both aerial- and liquid-phase conditions. Cereibacter sp. KGU-NF001 showed promising oxygen tolerance and was able to maintain hydrogen production of 1.33 mL/mg/d even when the atmosphere contained 12% oxygen. Our findings illustrate that biohydrogen production may be achieved by photosynthetic bacteria under oxygen-containing aerial-phase conditions, indicating a possible pathway to help lower our reliance on fossil fuels.

Purpose: To report anterior segment evaluation in patients with mucopolysaccharidosis 1 using anterior segment optical coherence tomography and in vivo confocal microscopy. Observations: Case 1 involved a 26-year-old man with mucopolysaccharidosis 1 Hurler–Scheie syndrome who presented with a corrected distance visual acuity of 20/67 in the right eye and 20/50 in the left eye because of bilateral corneal opacification. He had undergone bone marrow transplantation at the age of 7 years. Anterior segment optical coherence tomography revealed a very narrow anterior chamber angle; a flat, thickened cornea; and astigmatism with the rule. Confocal microscopy demonstrated hyper-refractive deposits in the corneal epithelial basal layer, decreased subepithelial nerves, and depletion of keratocytes in the anterior and posterior stroma. Endothelial cells were not visible.Case 2 involved an 18-year-old woman who presented with increased corneal opacity in both eyes. Her medical history included mucopolysaccharidosis 1 Hurler syndrome and bone marrow transplantation at the age of 2 years. Her corrected distance visual acuity could not be measured because of severe intellectual disability. Anterior segment optical coherence tomography demonstrated a flat and thick cornea. Conclusions and importance: Anterior segment optical coherence tomography can provide detailed morphological information for the anterior segment in patients with severe corneal opacification associated with mucopolysaccharidosis 1. Moreover, in vivo confocal microscopy facilitates the detailed observation of corneal cellular changes. Thus, these tools can provide data that would contribute to an increased understanding of corneal changes in patients with mucopolysaccharidosis 1.

We have already proposed a multiple-vehicle bike sharing system routing problem (mBSSRP) to adjust the number of bicycles at each port using multiple vehicles in short time. However, there are many strict constraints in the mBSSRP, thus it is difficult to obtain feasible solutions of the mBSSRP for some instances. To obtain feasible solutions of the mBSSRP, we have proposed a mBSSRP with soft constraints (mBSSRP-S) that removes some constraints from mBSSRP and appends violations to an objective function as penalties, and a searching strategy that explores both the feasible and infeasible solution spaces. Numerical experiments indicated that solving mBSSRP-S to obtain feasible solutions of mBSSRP results in better performance than solving mBSSRP directly. However, mBSSRP-S includes infeasible solutions of mBSSRP, thus the neighborhood solutions and computational costs increase. In this study, we propose search strategies with low computational costs while maintaining performance. In particular, we propose two search strategies: the first one is to reduce neighborhood solutions to obtain a feasible solution in a short time before finding a feasible solution of the mBSSRP, and the second one is to change the problem to be solved (mBSSRP or mBSSRP-S) after a feasible solution is obtained and to search good near-optimal solutions in a short time. As the first search strategy, we propose two search methods for reducing the number of neighborhood solutions in the Or-opt and the CROSS-exchange and compare their performance with our previous results. From numerical experiments, we confirmed that a feasible solution can be obtained within a short time by exploring only the normal order insertion of the Or-opt and the normal order exchange of the CROSS-exchange as the neighborhood solutions. Next, as the second search strategy after a feasible solution of mBSSRP is obtained, we propose four search methods and compare their performance with our previous results. Numerical experiments show that the search method that only searches for the normal order insertion of the Or-opt and the normal order exchange of the CROSS-exchange with hard constraints after obtaining a feasible solution can obtain short tours within a short time.

Sonic hedgehog (Shh) and its signaling have been identified in several human cancers, and increased levels of its expression appear to correlate with disease progression and metastasis. However, the role of Shh in bone destruction associated with oral squamous cell carcinomas, which frequently invade the maxilla or the mandible, is still unclear. In this study we show that the use of siRNA for Shh to block SHH secreted by SAS oral squamous cell carcinoma cells suppressed the tumor growth and tumor angiogenesis of subcutaneous SAS xenografts in vivo. Moreover, blockade of Shh in SAS cells decreased tumor growth and osteoclast number in a tibial metaphysis mouse model. Significantly, we clearly show that SHH stimulated osteoclast formation in a co-culture system consisting of murine bone stromal ST2 cells and murine CD11b(+) bone marrow cells. These findings suggest that Shh signaling is a potential target for the treatment of oral squamous cell carcinoma associated with bone destruction. [ABSTRACT FROM AUTHOR]

Purpose: To report successful medical management of Pythium insidiosum keratitis using an antibiotic combination of minocycline, linezolid, and chloramphenicol. Observations: A 20-year-old Japanese man was referred for visual disturbance, hyperemia, and discharge from his right eye. Slit-lamp examination revealed a paracentral corneal hyphate ulcer. His visual acuity was 20/28. Smear examination of corneal scrapings revealed a filamentous fungus. Pimaricin ointment four times a day and voriconazole eye drops hourly were initially prescribed. Although intravenous liposomal amphotericin B 100 mg was added, the corneal infiltrates and ulcer worsened. The possibility of P. insidiosum keratitis was considered, and in vitro antifungal susceptibility testing were performed based on the disc diffusion method. The inhibition zones around each antibiotic disc revealed that the pathogen was susceptible to minocycline, linezolid, and chloramphenicol. Therefore, minocycline ointment four times a day, chloramphenicol eye drops hourly, and linezolid 1200 mg orally per day were also administered. Eventually, sequencing of ribosomal DNA confirmed the pathogen to be P. insidiosum. The triple regimen dramatically improved the patient's keratitis. Therapeutic penetrating keratoplasty for corneal perforation was successfully performed, and his visual acuity recovered from 20/2000 to 20/25. Conclusions and importance: We have encountered a case of P. insidiosum keratitis that responded to a combination of minocycline, linezolid, and chloramphenicol. This triple combination should be considered in patients with P. insidiosum keratitis. Keywords: Pythium insidiosum keratitis, Minocycline, Linezolid, Chloramphenicol

Objectives The 1995 Great Hanshin-Awaji Earthquake had an enormous negative impact on survivors’ health. Many survivors experienced psychological distress and their long-term psychological recovery process remains unclear. Our objective was thus to explore this long-term psychological recovery process.Design Qualitative study.Setting From January to December 2015, face-to-face interviews were conducted in Kobe, Japan.Participants 20 affected survivors (55% female; ages ranged from 7 to 60 at the time of the disaster)—10 volunteer storytellers, six first responders (firefighters/public health nurses) and four post-traumatic stress disorder patients.Outcome measures We asked participants about their experiences and psychological distress relating to the earthquake and what facilitated or hindered their psychological recovery. We analysed interview data using constructivist grounded theory.Results Participants experienced diverse emotional reactions immediately after the disaster and often hyperfocused on what they should do now. This hyperfocused state led to both mental and physical health problems several months after the disaster. Months, and sometimes years, after the disaster, guilt and earthquake narratives (ie, expressing thoughts and feelings about the earthquake) played key roles in survivors’ psychological recovery: guilt suppressed their earthquake narrative; conversely, the narrative alleviated feelings of distress about the earthquake. In time, participants reconsidered their earthquake experiences both alone and through social interaction. This alleviated their emotional reactions; however, participants still experienced attenuated emotional reactions, and some hid their feelings of distress even 20 years postdisaster. Interpersonal relationships modified this psychological process both positively and negatively.Conclusions Future psychosocial support plans for disaster survivors may need to (1) include both mental and physical care in the transition from the acute phase to the recovery phase; (2) facilitate supportive interpersonal relationships for survivors during the mid-term recovery phase and (3) provide long-term psychological support to the most traumatised survivors, even if they appear to be functioning normally.

In bicycle sharing systems, many vehicles restore bicycles to ports. To construct the shortest tour of these vehicles, in a previous work, we formulated the multiple-vehicle bike sharing system routing problem (mBSSRP) and demonstrated that an optimal solution can be obtained for small-sized instances through a general-purpose mixed-integer linear programming solver. However, for large-sized instances, the optimal solution could not be found in a reasonable time frame. Therefore, to find near-optimal solutions for the mBSSRPs in a short time, in this study, we develop a method with a searching strategy, which explores both the feasible and infeasible solution spaces. To investigate the performance of the proposed method, we solve benchmark problems of mBSSRP. In addition, we compare the proposed method with the method exploring only the feasible solution space, in terms of performance. The results of the numerical experiments demonstrate that the proposed method can reach optimal solutions for almost all small-sized mBSSRP instances and that searching both the feasible and infeasible solution spaces yields good feasible solutions both for small-sized and large-sized instances.

Epstein-Barr virus (EBV) has been detected in several T- and NK-cell neoplasms such as extranodal NK/T-cell lymphoma nasal type, aggressive NK-cell leukemia, EBV-positive peripheral T-cell lymphoma, systemic EBV-positive T-cell lymphoma of childhood, and chronic active EBV infection (CAEBV). However, how this virus contributes to lymphomagenesis in T or NK cells remains largely unknown. Here, we examined NF-κB activation in EBV-positive T or NK cell lines, SNT8, SNT15, SNT16, SNK6, and primary EBV-positive and clonally proliferating T/NK cells obtained from the peripheral blood of patients with CAEBV. Western blotting, electrophoretic mobility shift assays, and immunofluorescent staining revealed persistent NF-κB activation in EBV-infected cell lines and primary cells from patients. Furthermore, we investigated the role of EBV in infected T cells. We performed an in vitro infection assay using MOLT4 cells infected with EBV. The infection directly induced NF-κB activation, promoted survival, and inhibited etoposide-induced apoptosis in MOLT4 cells. The luciferase assay suggested that LMP1 mediated NF-κB activation in MOLT4 cells. IMD-0354, a specific inhibitor of NF-κB that suppresses NF-κB activation in cell lines, inhibited cell survival and induced apoptosis. These results indicate that EBV induces NF-κB-mediated survival signals in T and NK cells, and therefore, may contribute to the lymphomagenesis of these cells.

[This corrects the article DOI: 10.1371/journal.pone.0174136.].

Background and Aim: Heat shock protein 90 (HSP90) and mammalian target of rapamycin (mTOR) are involved in the molecular pathogenesis of advanced oral squamous cell carcinoma. HSP90 inhibitors are capable of effectively interfering with multiple signaling pathways, including the mTOR signaling pathway. However, the combined effects of HSP90 and mTOR inhibitors on oral squamous cell carcinoma are still unknown. In this study, we investigated the dual treatment of the novel HSP90 inhibitor NVP-AUY922 and temsirolimus against oral squamous cell carcinoma., Materials and Methods: The effect of the combination of NVP-AUY922 and temsirolimus on oral squamous cell carcinoma in vitro and in vivo was determined by MTS assay and mouse xenograft models. The effect of the combination on angiogenesis was determined by tube formation assay and angioreactor., Results: The combination treatment of NVP-AUY922 and temsirolimus significantly inhibited the proliferation of SAS oral squamous cell carcinoma cells in vitro and suppressed the growth of oral squamous cell carcinoma xenografts in vivo. We have clearly shown that the combination treatment of NVP-AUY922 and temsirolimus inhibited vascular formation both in vitro and in vivo. Moreover, the combination treatment of NVP-AUY922 and temsirolimus prolonged the survival rate in mice xenografted with oral squamous cell carcinoma., Conclusions: Here, we showed the activity of a combination of mTOR and HSP90 inhibitors for the treatment of advanced oral squamous carcinoma.

Objectives: Focal adhesion kinase (FAK) overexpression is frequently found in invasive and metastatic cancers, but its role in oral squamous cell carcinoma is not yet well understood. In order to seek therapies targeting oral squamous cell carcinoma, we developed the novel FAK Tyr(397) inhibitor TAE226 and investigated its anti-tumor effects and mechanisms., Materials and Methods: Expression of phosphorylated FAK Tyr(397) was examined by immunohistochemical and immunoblot analysis. The effect of TAE226 on in vitro and in vivo studies were confirmed by proliferation, cell cycle, apoptosis and angiogenesis analysis., Results: We found that phosphorylated FAK was highly expressed in human tongue oral squamous cell carcinoma in patients. Importantly, TAE226 greatly suppressed the proliferation, migration and invasion of human oral squamous cell carcinoma SAS cells with an apparent structural change of actin fiber and a loss of cell adhesion. In addition, TAE226 inhibited the expression of phospho-FAK Tyr(397) and phospho AKT Ser(473), resulting in caspase-mediated apoptosis. Furthermore, oral administration of TAE226 in mice suppressed the growth and angiogenesis of oral squamous cell carcinoma xenografts in vivo., Conclusions: Our results provide compelling evidence that FAK is critically involved in oral squamous cell carcinoma and that the FAK inhibitor TAE226 can potentially be effectively used for the treatment of oral squamous cell carcinoma., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Focal adhesion kinase (FAK) is a 125-kDa non-receptor type tyrosine kinase that localizes to focal adhesions. FAK overexpression is frequently found in invasive and metastatic cancers of the breast, colon, thyroid, and prostate, but its role in osteolytic metastasis is not well understood. In this study, we have analyzed anti-tumor effects of the novel FAK Tyr(397) inhibitor TAE226 against bone metastasis in breast cancer by using TAE226. Oral administration of TAE226 in mice significantly decreased bone metastasis and osteoclasts involved which were induced by MDA-MB-231 breast cancer cells and increased the survival rate of the mouse models of bone metastasis. TAE226 also suppressed the growth of subcutaneous tumors in vivo and the proliferation and migration of MDA-MB-231 cells in vitro. Significantly, TAE226 inhibited the osteoclast formation in murine pre-osteoclastic RAW264.7 cells, and actin ring and pit formation in mature osteoclasts. Moreover, TAE226 inhibited the receptor activator for nuclear factor κ B Ligand (RANKL) gene expression induced by parathyroid hormone-related protein (PTHrP) in bone stromal ST2 cells and blood free calcium concentration induced by PTHrP administration in vivo. These findings suggest that FAK was critically involved in osteolytic metastasis and activated in tumors, pre-osteoclasts, mature osteoclasts, and bone stromal cells and TAE226 can be effectively used for the treatment of cancer induced bone metastasis and other bone diseases., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The mammalian target of rapamycin (mTOR) is engaged in the molecular pathogenesis of oral squamous cell carcinoma, which frequently invades the maxilla or the mandible. However, the effects of a mTOR inhibitor on bone destruction associated with oral squamous cell carcinoma are still unclear. In this study, we investigated the antitumor effect of temsirolimus-mediated mTOR inhibition against advanced oral squamous cell carcinoma. Temsirolimus inhibited the proliferation and migration of HSC-2 oral squamous cell carcinoma cells in vitro and suppressed the growth of oral squamous cell carcinoma xenografts in vivo. Significantly, we clearly show that temsirolimus inhibited osteoclast formation both in vitro and in vivo. Reverse transcriptase-PCR analysis showed that temsirolimus decreased the mRNA expression of receptor activator for nuclear factor-κB ligand, known as an osteoclast differentiation factor in bone stromal ST2 cells. Moreover, temsirolimus normalized blood-free calcium concentration in mouse models for humoral hypercalcemia. These findings suggest that mTOR signaling is a potential target of oral squamous cell carcinoma associated with bone destruction, and hence we describe the efficacy of temsirolimus for the treatment of advanced oral squamous carcinoma., (©2010 AACR.)

Background: Gingival squamous cell carcinoma (SCC) cells frequently invade mandibular bone, and this destruction is associated with a worse prognosis. However, the relationship between bone destruction and associated factors is unclear. In this study, the role and diagnostic utility of transforming growth factor-beta (TGF-beta) type I receptor (TbetaRI) in bone destruction of the mandible was investigated., Patients and Methods: The expression of TbetaRI was explored by using an immunohistochemical method on paraffin-embedded tissues from 21 cases of mandibular SCC. An inhibitor of the kinase activity of the TbetaRI (TbetaRI-I) was used to assess the role of TbetaRI in bone destruction by a human oral SCC cell line (HSC-2) that highly expresses TbetaRI., Results: TbetaRI-positive signals were closely associated with destructive invasion of the mandible by oral SCC cells. Consistent with these results, TbetaRI-I greatly reduced HSC-2 cell-induced bone destruction and osteoclast formation in vivo and in vitro. TbetaRI-I treatment reduced the expression of TNF-alpha, RANKL and connective tissue growth factor (CTGF/CCN2), all of which were up-regulated by TGF-beta in HSC-2 cells., Conclusion: These data demonstrated an important role for TGF-beta signalling in bone invasion by oral SCC cells, and suggest that the bone destruction is mediated by RANKL, TNF-alpha and CCN2.