Clarification of the Mechanism of Inhibiting the Maillard Reaction between Phosphatidylethanolamine and Sugars by Adding Fatty Acid Calcium Salts.

Lecithin is known to undergo heat induced deterioration by the Maillard reaction between 1 mol of any sugar, except 2-deoxy sugar, and 2 mol of phosphatidylethanolamine (PE). However, we have previously reported that adding fatty acid metal salts can inhibit heat deterioration of soybean lecithin (SL). To clarify the mechanism of inhibition, 1,2-di-O-stearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE), d-glucose, and calcium stearate or calcium decanoate were heated in octane. When DSPE and d-glucose with calcium stearate or calcium decanoate were heated in the octane, the heat deterioration of DSPE was significantly inhibited, and no increase in UV absorption at 350 nm was observed. From these reactant solutions, one compound having a phosphate group and no primary amine was isolated, and NMR spectra confirmed that two molar of stearic acid derived from DSPE were coordinated to the amino group and phosphate group of DSPE. Therefore, we concluded that adding fatty acid metal salts reduced the nucleophilic reactivity of the amino group of PE and inhibited the Maillard reaction with sugars because two molar of fatty acid derived from PE coordinated to the amino group and phosphate group of PE.

A rare etiology of mild encephalitis/encephalopathy with reversible splenial lesion.

Legionella is a rare cause of mild encephalitis/encephalopathy with reversible splenial lesion, which should be considered in patients with risk factors. Brain magnetic resonance imaging (MRI) and legionella urinary antigen test can help the diagnosis since cerebrospinal fluid (CSF) can be normal.

Use of Fatty Acid Metal Salts for Preventing Maillard Reaction-Driven Browning of Lecithins.

We previously reported that fluid soybean lecithin (SL) undergoes heat deterioration by the newly reported pseudo-Maillard rearrangement reaction. To inhibit heat deterioration, SLs were treated with metal silicates, such as magnesium silicate and calcium silicate. When soybean fatty acid was added to SL before treatment with calcium silicate, the color index after heating improved significantly as the acid value increased from 10 to 35 mg KOH/g. To elucidate the role of soybean fatty acid, calcium silicate treatment was carried out by adding several fatty acids to SL. Although saturated fatty acids had no effect on the heat deterioration of SL, unsaturated fatty acids were significantly more effective at inhibiting heat deterioration. Furthermore, for unsaturated fatty acids, it was confirmed that the calcium concentration increased in SL. Based on these results, several fatty acid metal salts were added to confirm whether heat deterioration while heating SL could be inhibited. It was observed that the heat deterioration of SL could be inhibited with fatty acid metal salts, regardless of whether the fatty acids were saturated or unsaturated and whether the metal was monovalent, divalent, or trivalent. Therefore, in this study, we clarified that the heat deterioration of SL could be inhibited by adding fatty acid metal salts to SL. Among sodium stearate, calcium stearate, magnesium stearate, barium stearate, and aluminum tristearate, the divalent fatty acid metal salts had a stronger inhibitory effect on heat deterioration than the monovalent and trivalent salts.

10-Gingerol Suppresses Osteoclastogenesis in RAW264.7 Cells and Zebrafish Osteoporotic Scales.

Osteoporosis is the most common aging-associated bone disease and is caused by hyperactivation of osteoclastic activity. We previously reported that the hexane extract of ginger rhizome [ginger hexane extract (GHE)] could suppress receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. However, the anti-osteoclastic components in GHE have not yet been identified. In this study, we separated GHE into several fractions using silica gel column chromatography and evaluated their effects on osteoclastogenesis using a RAW264.7 cell osteoclast differentiation assay (in vitro) and the zebrafish scale model of osteoporosis (in vivo). We identified that the fractions containing 10-gingerol suppressed osteoclastogenesis in RAW264.7 cells detected by tartrate-resistant acid phosphatase (TRAP) staining. In zebrafish, GHE and 10-gingerol suppressed osteoclastogenesis in prednisolone-induced osteoporosis regenerated scales to promote normal regeneration. Gene expression analysis revealed that 10-gingerol suppressed osteoclast markers in RAW264.7 cells [osteoclast-associated immunoglobulin-like receptor, dendrocyte-expressed seven transmembrane protein, and matrix metallopeptidase-9 (Mmp9)] and zebrafish scales [osteoclast-specific cathepsin K (CTSK), mmp2, and mmp9]. Interestingly, nuclear factor of activated T-cells cytoplasmic 1, a master transcription regulator of osteoclast differentiation upstream of the osteoclastic activators, was downregulated in zebrafish scales but showed no alteration in RAW264.7 cells. In addition, 10-gingerol inhibited CTSK activity under cell-free conditions. This is the first study, to our knowledge, that has found that 10-gingerol in GHE could suppress osteoclastic activity in both in vitro and in vivo conditions.

Lecithin-Based Dermal Drug Delivery for Anti-Pigmentation Maize Ceramide.

Ceramides have several well-known biological properties, including anti-pigmentation and anti-melanogenesis, which make them applicable for use in skincare products in cosmetics. However, the efficacy of ceramides is still limited. Dermal or transdermal drug delivery systems can enhance the anti-pigmentation properties of ceramides, although there is currently no systemic evaluation method for the efficacy of these systems. Here we prepared several types of lecithin-based emulsion of maize-derived glucosylceramide, determining PC70-ceramide (phosphatidylcholine-base) to be the safest and most effective anti-pigmentation agent using zebrafish larvae. We also demonstrated the efficacy of PC70 as a drug delivery system by showing that PC70-Nile Red (red fluorescence) promoted Nile Red accumulation in the larval bodies. In addition, PC70-ceramide suppressed melanin in mouse B16 melanoma cells compared to ceramide alone. In conclusion, we developed a lecithin-based dermal delivery method for ceramide using zebrafish larvae with implications for human clinical use.

A single-arm phase II study of nab-paclitaxel for patients with chemorefractory non-small cell lung cancer.

BACKGROUND: We aimed to evaluate the efficacy and safety of nab-paclitaxel in patients with refractory advanced non-small cell lung cancer who failed previous chemotherapy. METHODS: Patients were required to have an Eastern Cooperative Oncology Group performance status of 0-2 and adequate organ function. Patients received nab-paclitaxel, 100 mg/m2 i.v. on days 1, 8, and 15 every 4 weeks. The primary endpoint was the overall response rate. Secondary endpoints were the progression-free survival time, overall survival, and the toxicity profile. RESULTS: From July 2013 to July 2015, a total of 31 patients were enrolled. Fourteen patients received nab-paclitaxel as a second-line and 17 received it as an over third-line therapy. Each patient received a median of 5 treatment cycles (range, 1-11). The overall response rate was 19.3% (95% confidence interval, 9.1-36.2%) (complete response (n = 0), partial response (n = 6), stable disease (n = 17), and progressive disease (n = 8)). The median progression-free survival time was 4.5 months (95% confidence interval 3.5-6.3 months), median overall survival time was 15.7 months, and 1-year survival rate was 54.8%. Most common grade 3 or 4 non-hematological toxicities were elevated aspartate transaminase level (3.2%) and sensory neuropathy (9.6%). Neutropenia was the most common grade 3 or 4 adverse events (38.6%), and febrile neutropenia developed in 12.9% patients. No treatment-related deaths were observed in this study. CONCLUSION: Primary endpoint was met. Single agent nab-paclitaxel showed significant clinical efficacy and manageable toxicities for patients with chemorefractory advanced non-small cell lung cancer even if late line setting. TRIAL REGISTRATION: UMIN000011696 . The date of registration was July 11th, 2013.

Clinical application of immunocytochemical detection of ALK rearrangement on cytology slides for detection or screening of lung adenocarcinoma.

UNLABELLED: Immunohistochemical screening of Anaplastic lymphoma kinase (ALK) rearrangement has been regarded essential and routinely carried out to select treatment for lung adenocarcinoma. However, difficulty to approach a tumor by transbronchial lung biopsy (TBLB), it often fails to obtain tumor tissues whereas tumor cells are contained in cytology specimens simultaneously obtained when the bronchoscopy is done. Therefore we evaluated the expression of ALK protein by using immunohistochemistry (IHC) on TBLB specimens and immunocytochemistry (ICC) on brushing smear cytology slides in the same cases, and compared the concordance rate of IHC and ICC results. ICC was carried out on routine Papanicolau-stained slides after cytology diagnosis and decolorization. RESULTS: Eighteen patients with adenocarcinoma were extracted in the Hirosaki University Hospital and the Hirosaki National Hospital. IHC and ICC results showed a very high concordance rate: sensitivity of ICC in comparison with IHC was 85.7% (6/7), specificity was 100% (11/11), positive predictive value was 100% (6/6), and negative predictive value was 91.6% (11/12). Detection of ALK rearrangement using ICC on routine Papanicolau cytology slides is considered to be advantageous for lung cancer treatments.

The long-term survival of a thymic carcinoma patient treated with S-1: a case report and literature review.

BACKGROUND: Thymic carcinoma is a rare neoplasm of the thymus. Systemic chemotherapy is an important therapeutic modality for thymic carcinoma. However, no standard chemotherapy for this carcinoma has yet been established. The usefulness of second-line or later-line chemotherapy has remained unclear. A case of relapsed thymic carcinoma that was successfully treated by S-1 as second-line chemotherapy is reported herein. CASE PRESENTATION: A 73-year-old man diagnosed as having thymic carcinoma was treated with three cycles of first-line chemotherapy with ADOC (cisplatin, doxorubicin, vincristine, and cyclophosphamide) and additional radiotherapy (50 Gy). Since his serum cytokeratin 19 fragment level increased suddenly after 3 months of stable disease, he was considered to have progressive disease, and was given S-1 as chemotherapy. Two months later, he had partial response, and the S-1 treatment has been continued since July 2009. Progression-free survival of greater than 4 years was obtained with S-1. CONCLUSION: A case of relapsed thymic carcinoma that was treated with S-1, and continues to show a long progression-free survival with good quality of life on treatment is described. S-1 might be an active agent against relapsed thymic carcinoma.

A case of lung adenocarcinoma harboring EGFR mutation and EML4-ALK fusion gene.

BACKGROUND: Lung cancer is the leading cause of cancer-related death worldwide. Epidermal growth factor receptor (EGFR)--tyrosine kinase inhibitor (TKI) is used for the patients with EGFR-mutant lung cancer. Recently, phase III studies in the patients with EGFR-mutant demonstrated that EGFR-TKI monotherapy improved progression-free survival compared with platinum-doublet chemotherapy. The echinoderm microtubule-associated protein-like 4 (EML4)--anaplastic lymphoma kinase (ALK) fusion oncogene represents one of the newest molecular targets in non-small cell lung cancer (NSCLC). Patients who harbor EML4-ALK fusions have been associated with a lack of EGFR or KRAS mutations. CASE PRESENTATION: We report a 39-year-old patient diagnosed as adenocarcinoma harboring EGFR mutation and EML4-ALK fusion gene. We treated this patient with erlotinib as the third line therapy, but no clinical benefit was obtained. CONCLUSION: We experienced a rare case with EGFR mutation and EML4-ALK. Any clinical benefit using EGFR-TKI was not obtained in our case. The therapeutic choice for the patients with more than one driver mutations is unclear. We needs further understanding of the lung cancer molecular biology and the biomarker information.

[A case of adenocarcinoma of the lung with a long-term therapeutic effect due to S-1 administered as the sixth regimen].

The patient was a 36-year-old male. He visited our department in February 2004 as a prior chest X-ray revealed multiple nodular shadows. After further examinations, he was diagnosed with stage IV adenocarcinoma of the lung. As treatment, 5 courses of carboplatin (CBDCA)+docetaxel (DOC), 3 courses of CBDCA+gemcitabine (GEM), 6 weeks of gefitinib, and 3 courses of GEM were administered. However, the tumor progressed, and S-1 (120 mg/day, 2 weeks on, 1 week off) was administered as the sixth regimen from May 2006. No severe side effects were observed, and an antitumor action was achieved over a relatively long period. Therefore, it was suggested that a single administration of S-1 for non-small cell lung cancer, which had been treated with other multiple regimens, is safe, and long-term control of the disease can be expected.

Heat deterioration of phospholipids. VI. Clarification of mechanism for the new pseudo Maillard rearrangement reaction by using 2-aminoethyl dihydrogenphosphate.

2,3-Dihydro-1H-imidazo[1,2-a]pyridine-4-ylium derivatives with UV absorption at 350 nm were formed by reaction of one molar of any sugar except 2-deoxysugars with two molar of phosphatidylethanolamines involving a new pseudo Maillard rearrangement reaction. To elucidate the reaction mechanism, 2-aminoethyl dihydrogenphosphate, which had a partially similar structural moiety to PE, was reacted with D-galactose. Though the UV absorption at 365 nm was not observed and the four pyridinium derivatives were also not formed in the reactant solution, the UV absorption at 286 nm and browning of the reactant solution were observed. From this reactant solution, two compounds with lambdamax at 283 nm and 297 nm were isolated and former was determined as 5-(hydroxymethyl)furfural (HMF) and later did as phosphoric acid mono{2-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]ethyl}ester (PMPEE), which is a new compound, respectively. Because reaction of PMPEE with PE leads to form the pyridinium derivatives, we concluded that a compound like PMPEE was one of intermediates in this new reaction.

Heat deterioration of phospholipids. V. A new rearrangement reaction of sugars and phosphatidylethanolamine.

Novel four 2,3-dihydro-1H-imidazo[1,2-a]pyridine-4-ylium derivatives were obtained with increase of UV absorption at 350 nm and browning of the solution by heating paste lecithin from soybean (SL) in octane. These four derivatives were formed by reaction of one molar of any sugar except 2-deoxysugars with two molar of phosphatidylethanolamines (PE) in SL. To confirm the reaction mechanism, several (13)C-labeled-sugars were reacted with 1,2-di-O-stearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE), respectively. These reactants clearly showed that five carbons of the pyridinium ring and one carbon of the substituted group were based on those of a sugar and that the formation of the pyridinium derivatives was accompanied with cleaving between the carbons of 1- and 2-positions in the sugar and rearrangement. This reaction is a new rearrangement reaction and we named it "new pseudo Maillard rearrangement reaction".

Level of antibodies against mycobacterial glycolipid in the effusion for diagnosis of tuberculous pleural effusion.

BACKGROUND: Diagnosing tuberculous pleural effusion (pTB) is often difficult because the culturing of tubercle bacilli results in a negative test in the majority of cases. Serological tests for the detection of antibodies to tuberculous glycolipid (TBGL) and lipoarabinomannan (LAM) have been introduced for the diagnosis of pulmonary tuberculosis. We examined the levels of these antibodies, adenosine deaminase (ADA) and interferon-gamma (IFN-gamma) in the pleural effusion and compared their diagnostic values in pTB. METHODS: We studied 65 patients with pleural effusion. Of those, 19 patients were diagnosed as having pTB according to our broad case definition. The etiologies in the other 46 patients were malignant effusion, transdative effusion and miscellaneous diseases. Determiner TBGL antibody (D-TBGL-Ab) and MycoDot were used for the detection of anti-LAM and anti-TBGL antibodies, respectively, in the pleural effusion. RESULTS: The sensitivity of ADA was 78.9% (15/19) and the specificity 97.8% (45/46). The sensitivity of IFN-gamma was 84.2% (16/19) and the specificity 93.5% (43/46). The sensitivities of D-TBGL-Ab and MycoDot were both 52.6% (10/19) and their specificities were 95.7% (44/46) and 97.8% (45/46), respectively. When DTBGL-Ab (cutoff point: 2.0 U/ml) and ADA activity (cutoff point: 57 IU/l) were combined, the sensitivity was 94.7% (18/19) and the specificity 93.5% (43/46). CONCLUSIONS: In the diagnosis of pTB, D-TBGL-Ab and MycoDot each have low sensitivity but high specificity. When D-TBGL-Ab is used in combination with ADA, the sensitivity and specificity are both >90%. We conclude that D-TBGL-Ab and ADA in combination are useful in the diagnosis of pTB.

The significance of cathepsins, thrombin and aminopeptidase in diffuse interstitial lung diseases.

To determine the significance of proteases in interstitial lung diseases, we examined the activity of cathepsins, thrombin, and aminopeptidase in bronchoalveolar lavage (BAL) fluid from patients with these disorders. Significantly increased activities of cathepsin H and aminopeptidase were detected in BAL fluid from patients with idiopathic pulmonary fibrosis (IPF), cryptogenic organizing pneumonia (COP), chronic eosinophilic pneumonia (CEP) and hypersensitivity pneumonitis (HP). Significantly higher activity of cathepsin B was found in BAL fluid from patients with CEP. The activity of thrombin was significantly higher in patients with IPF and CEP. In patients with IPF, there were significant correlations between neutrophil number and the activity of cathepsin B, cathepsin H or aminopeptidase. In patients with COP and HP, the activity of the proteases was significantly higher in patients with higher number of lymphocytes than in those with lower number of lymphocytes. The present study suggests that the activity of the proteases is a useful marker in activity of the interstitial lung diseases, and may have a role in the pathogenesis of these disorders.

Vascular endothelial growth factor level as a prognostic determinant of small cell lung cancer in Japanese patients.

OBJECTIVE: To clarify the clinical significance of vascular endothelial growth factor (VEGF) in Japanese patients with small cell lung cancer (SCLC). MATERIALS AND METHODS: We measured serum VEGF levels using an enzyme-linked immunosorbent assay in 45 patients with SCLC before treatment and in 38 patients with benign pulmonary disease and in 32 healthy subjects (71 non-malignant subjects). VEGF immunostaining was performed in tissue biopsies obtained from 23 SCLC patients during bronchoscopic examination. RESULTS: Median serum VEGF level was 332 pg/ml in patients with SCLC and 160 pg/ml in non-malignant subjects, respectively. The 95% cut-off level to exclude non-malignant subjects was 500 pg/ml. An elevated VEGF level (>500 pg/ml) was found more frequently in patients with extensive disease of SCLC than in those with the limited disease (p<0.01). A significant positive correlation was found between the serum VEGF level and platelet count in SCLC patients (r=0.389; p=0.0083). Serum VEGF level also correlated with serum lactate dehydrogenase in SCLC patients (r=0.381; p=0.0098). However, it did not correlate with serum neuron-specific enolase and pro-gastrin-releasing peptide level. Patients with the elevated VEGF levels had significantly shorter progression-free time than those with the normal VEGF levels (p<0.05). Patients with the elevated VEGF levels had a significantly shorter overall survival time than those with the normal VEGF levels in univariate survival analysis (p<0.05). Further, the elevated VEGF level remained as a significant determinant of poor survival in multivariate analysis (p<0.01). Serum VEGF level was significantly higher in patients with positive VEGF protein immunoreactivity in tumor tissue in SCLC. CONCLUSION: Elevated serum VEGF levels were associated with poor outcome in SCLC.

Transforming growth factor beta stimulates rheumatoid synovial fibroblasts via the type II receptor.

Transforming growth factor (TGF)-beta regulates the function of fibroblasts, and has been shown to have a role in the pathogenesis of rheumatoid arthritis (RA) because several studies have demonstrated the presence of TGF-beta in the synovial tissue and synovial fluids of RA patients. In this study, we examined the expression of TGF-beta receptors in synovial fibroblasts of patients with RA and demonstrated the significance in functional responses of synovial fibroblasts to TGF-beta in this disorder. Transforming growth factor beta1 stimulated the expression of connective tissue growth factor (CTGF) in fibroblasts of patients with RA more than in those of patients with osteoarthritis (OA). Transforming growth factor beta1 induced the chemotactic migration of RA synovial fibroblasts and inhibited their proliferation significantly more than OA synovial fibroblasts. Both RA and OA synovial fibroblasts expressed detectable amounts of TGF-beta receptor type II mRNA, but the expression was higher in RA patients than in OA patients, as assessed by reverse transcriptase-polymerase chain reaction. There was no significant difference in the expression of TGF-beta receptor type I or type III in synovial fibroblasts between RA and OA patients. These results indicate that synovial fibroblasts of RA patients express the increased TGF-beta receptor type II, which is associated with altered responses to TGF-beta observed in CTGF expression, chemotaxis, and proliferation of RA synovial fibroblasts, and may have an important role in the pathogenesis of RA.