Development of a quantitative analysis method for assessing patient body surface deformation using an optical surface tracking system.

This study aimed to develop a new method to quantitatively analyze body shape changes in patients during radiotherapy without additional radiation exposure using an optical surface tracking system. This method's accuracy was evaluated using a cubic phantom with a known shift. Surface images of three-dimensionally printed phantoms, which simulated the head and neck shapes of real patients before and after treatment, were used to create a deformation surface area histogram. The near-maximum deformation value covering an area of 2 cm2 in the surface image (Def-2cm2) was calculated. A volumetric modulated arc therapy (VMAT) plan was also created on the pre-treatment phantom, and the dose distribution was recalculated on the post-treatment phantom to compare the dose indices. Surface images of four patients were analyzed to evaluate Def-2cm2 and examine whether this method can be used in clinical cases. Experiments with the cubic phantom resulted in a mean deformation error of 0.08 mm. With head and neck phantoms, the Def-2cm2 value was 17.5 mm, and the dose that covered 95% of the planning target volume in the VMAT plan decreased by 11.7%, indicating that deformation of the body surface may affect the dose distribution. Although analysis of the clinical data showed no clinically relevant deformation in any of the cases, slight skin sagging and respiratory changes in the body surface were observed. The proposed method can quantitatively and accurately evaluate the deformation of a body surface. This method is expected to be used to make decisions regarding modifications to treatment plans.

Production of chitin nanoparticles by bottom-up approach from alkaline chitin solution.

Most of the series of nanochitins have been produced by the break-down process. In this study, chitin nanoparticles were prepared by a bottom-up process. Chitin was treated with sodium hydroxide to obtain an alkaline chitin aqueous solution. The alkaline chitin was regenerated by neutralization and then vigorously stirred to obtain chitin nanoparticles. The average particle size of the chitin nanoparticles was 7 nm. The individual particles were stably dispersed in water. Chitin nanoparticles had lower crystallinity than the raw material chitin and the surface of the chitin nanoparticles regenerated in water were presumed to be hydrophilic. The low crystallinity and the high hydrophilicity of the surface contributed to the high dispersibility of the chitin nanoparticles in water. Chitin nanoparticles had higher heat resistance than the raw material chitin, suggesting a large change in the higher-order structure associated with dissolution and subsequent regeneration of chitin. Since chitin nanoparticles interact with each other less than chitin nanofibers produced by mechanical treatment, the viscosity of nanoparticles was smaller than that of nanofibers. Therefore, it can be prepared at a high concentration. In addition, the chitin nanoparticles can be easily redispersed in water after being concentrated by centrifugation.

Usefulness of Liver Uptake Rate Constant in 99mTc-GSA Scintigraphy for the Risk Stratification of Patients Undergoing Hepatectomy: A New Method for Calculation.

Introduction: The use of technetium 99m diethylenetriaminepentaacetic acid-galactosyl human serum albumin (99mTc-GSA) scintigraphy parameters, HH15 and LHL15, in assessing the future liver remnant function is not expedient because of their nonlinear behaviour against liver volume. Uptake rate constant for the binding of 99mTc-GSA to asialoglycoprotein receptors is probably more favourable, but the reported calculation methods are complex. We devised a simple method to calculate the uptake rate constant, KrGSA. Methods: Radioactivity counts for the entire liver and heart regions were extracted at 10, 20, and 30 min. Using whole liver and heart volumes measured from single-photon emission computed tomography images, free radioactivity corresponding to the liver blood pool was subtracted. The time activity curve was fitted to the equation L(t) = L(∞) × [1 - Exp (-kt)] using Microsoft Office Excel (add-in free programme Solver)®, where L(∞) is the count at plateau level and k denotes KrGSA. Results: KrGSA values accurately identified liver cirrhosis and were similar to the KICG. The areas under the curve for KrGSA and KICG in the receiver operating characteristic analysis were 0.808 and 0.795, respectively, and a good correlation was seen between KrGSA and KICG. Discussion/Conclusion: KrGSA can be utilized as an alternative to KICG in assessing the future liver remnant function.

[A Case of Xanthogranulomatous Cholecystitis That Is Difficult to Distinguish from Advanced Gallbladder Cancer with Invasion of the Transverse Colon].

A 53-year-old woman was admitted to our hospital because of hepatic dysfunction found during a medical checkup. Cholecystitis was suspected, and unenhanced computed tomography (CT) was initially performed because she had bronchial asthma. However, a tumor-like lesion was seen at the bottom of the gallbladder. Contrast-enhanced CT was performed 3 weeks later, and the tumor-like lesion was enhanced and had increased in size. Endoscopic ultrasound fine-needle aspiration did not reveal any signs of malignancy. Colonoscopy revealed ulcerations in the transverse colon, and invasion from gallbladder cancer was suspected. Our preoperative diagnosis was xanthogranulomatous cholecystitis, but gallbladder cancer could not be excluded. Gallbladder bed resection and partial resection of the transverse colon were performed. Intraoperative frozen section analysis did not reveal any malignant findings; hence, we considered that lymph node dissection was unnecessary. Pathological examination confirmed xanthogranulomatous cholecystitis with abscess formation. In cases of surgery for xanthogranulomatous cholecystitis, it is important to consider that this condition could coexist with gallbladder cancer.

A novel hydrochloride-free chitosan oligosaccharide production method to improve taste.

Chitosan oligosaccharide hydrochloride (COS-HCl) has an unpleasant taste. To improve this taste, we used an enzymatic hydrolysis with chitosan to manufacture hydrochloride-free chitosan oligosaccharide (HFCOS). We found that HFCOS powder with weight-average molecular weight (Mw) of approximately 1000 and a high degree of deacetylation can be obtained from amorphous chitosan in carbonated water by enzymatic hydrolysis because amorphous chitosan slightly dissolves in carbonated water. HFCOS was stable enough to be atomized by spray-drying. However, HFCOS yield was quite low (15.7%). There were few chloride ions in HFCOS (0.1% ±â€¯0.1%), in contrast to the number of chloride ions in COS-HCl (16.6% ±â€¯0.6%) with statistical significance (p < 0.01). The molar ratio of anion/amino group of HFCOS was only 0.05. Therefore, it appeared that the amino groups of HFCOS were mostly free bases. In sensory evaluations, 5% w/w HFCOS was significantly sweeter than 5% w/w COS-HCl (p < 0.01); saltiness, acidity, bitterness were significantly weak (p < 0.01); and aftertaste and overall taste were significantly improved (p < 0.01). These results suggest that HFCOS can be obtained by a novel method of enzymatically hydrolyzing amorphous chitosan dissolved in carbonated water and this is one way to improve its taste.

Time-dependent inhibition (TDI) of CYP1A2 by a CYP3A4-mediated reactive metabolite: proposal for a novel mechanism of irreversible TDI by a non-suicide substrate.

1. The purpose of this study was to clarify the mechanism of DSP-1053 time-dependent inhibition (TDI) for CYP1A2. 2. DSP-1053 inhibited time- and concentration-dependently CYP1A2 activity in human liver microsomes even in a dilution assay. However, DSP-1053 was not metabolized by recombinant human CYP1A2. These findings indicate that the inhibitory effect of DSP-1053 on CYP1A2 does not follow a general mechanism-based inhibition (MBI) because it did not seem to be a suicide substrate. 3. In fact, CYP1A2 was not inhibited with DSP-1053 pre-incubation in recombinant human CYP1A2. On the other hand, CYP1A2 was potently inhibited after pre-incubation with DSP-1053 in a mixture of human recombinant CYP1A2 and CYP3A4. In addition, DSP-1053 TDI of CYP1A2 in human liver microsomes was drastically reduced not only by addition of a CYP3A4 inhibitor, but also by addition of potassium cyanide (KCN), which is a trapping agent for iminium ions. We also confirmed in this study that CYP1A2 suicide inhibition by DSP-1053 metabolites generated by CYP3A4 had only minimal role in DSP-1053 TDI of CYP1A2. 4. In conclusion, a possible mechanism for DSP-1053 TDI of CYP1A2 is that DSP-1053 iminium ion, which is generated by CYP3A4, departs from CYP3A4 without inhibiting it and covalently binds to CYP1A2.

Solubility-based Separation and Purification of Long-Chain Chitin Oligosaccharides with an Organic-Water Mixed Solvent.

A simple and rapid method for separation and purification of chitin oligosaccharides, (GlcNAc)n, with n ≥ 5 is presented. A commercially available chitin oligosaccharides sample, consisting of (GlcNAc)n with n = 1 - 7, was used as the starting material. Ten milligrams of the material was mixed with 100 µL of the 1 mol/L HCl. All the (GlcNAc)n species were dissolved in the aqueous medium. The aqueous solution was mixed with 900 µL of EtOH; the mixture was centrifuged, and the supernatant was removed to obtain a precipitate. The precipitate was found to consist mainly of (GlcNAc)n with n ≥ 5, indicating the significant difference in solubility between the short-chain (GlcNAc)n species with n ≤ 3 and the longer ones. By the repetition of the operations, a high purity long-chain (GlcNAc)n sample with n ≥ 5 could be prepared successfully. Since the long-chain (GlcNAc)n species are known to have excellent elicitor activity, this sample would be useful in the study of plant pathology, as well as chitin and chitosan chemistry.

[A Case of Heterochronic Ovarian Metastasis from Sigmoid Colon Cancer after Sigmoidectomy Treated with CapeOX That Included Bevacizumab].

The patient was a 41-year-old woman. When she was 39 years old, she had undergone laparoscopic high anterior resection for sigmoid colon cancer without adjuvant chemotherapy. Histologically, the surgical specimen was type 2, tub2, pT4a (SE), pN0, int, INF b, ly1, v1, and pStage II. Nine months after the operation, she suffered from abdominal fullness. Laborato- rydata showed elevation of tumor markers: the CEA level was 6.48 ng/mL, the CA19-9 level was 89.70 U/mL, and the CA125 level was 662 U/mL. Computed tomographyrevealed bilateral ovarian tumors and lung and peritoneal nodules with massive ascites. Chemotherapywas started with a regimen consisting of capecitabine plus oxaliplatin(CapeOX)that included bevacizumab. After 4 courses, the sizes of the lung and peritoneal nodules had decreased and the amount of ascites was almost zero. However, the ovarian tumors had increased in size and her sense of abdominal fullness had not improved. Bilateral oophorectomy with hysterectomy was performed to alleviate her symptom. Immunohistochemically, the resected ovarian tumors were negative for cytokeratin 7 and positive for cytokeratin 20. CapeOX with bevacizumab was then resumed. However, the lung tumor had graduallyincreased in size, and therefore, she underwent partial resection of the lung for the metastatic lung tumor.

Alginate gels with a combination of calcium and chitosan oligomer mixtures as crosslinkers.

Alginates are polysaccharides that are widely used in relation to their ability to form gels. Recently we reported that alginates may also form gels with chitosan oligomers as crosslinkers (Khong, Aarstad, Skjåk-Bræk, Draget, & Vårum, 2013). The purpose of the present study was to characterize alginate gels crosslinked with calcium and chitosan oligomers. Using two different alginates of similar molecular weights but different chemical composition, i.e. guluronic acid content of 46 and 68%, we found that both alginates could form homogeneous gels with calcium and chitosan oligomers separately and without syneresis. Systematic combinations of calcium and chitosan oligomers as crosslinkers were tested, showing that up to 50% of the calcium could be substituted with chitosan oligomers without reduction in gel strength or increased syneresis for the alginate with the lowest guluronic acid content. Furthermore, the kinetics of the combined gels were different from pure calcium alginate gels.

Metabolomic Analysis of Blood Plasma after Oral Administration of N-acetyl-d-Glucosamine in Dogs.

N-acetyl-d-glucosamine (GlcNAc) is a monosaccharide that polymerizes linearly through (1,4)-ß-linkages. GlcNAc is the monomeric unit of the polymer chitin. GlcNAc is a basic component of hyaluronic acid and keratin sulfate found on the cell surface. The aim of this study was to examine amino acid metabolism after oral GlcNAc administration in dogs. Results showed that plasma levels of ectoine were significantly higher after oral administration of GlcNAc than prior to administration (p < 0.001). To our knowledge, there have been no reports of increased ectoine concentrations in the plasma. The mechanism by which GlcNAc administration leads to increased ectoine plasma concentration remains unclear; future studies are required to clarify this mechanism.

Anti-inflammatory effects of orally administered glucosamine oligomer in an experimental model of inflammatory bowel disease.

Anti-inflammatory effects of oral administration of the glucosamine oligomers (chito-oligosaccharides: COS) were evaluated in an experimental model of inflammatory bowel disease (IBD). Oral administration of COS improved shortening of colon length and tissue injury (as assessed by histology) in mice. Oral administration of COS inhibited inflammation in the colonic mucosa by suppression of myeloperoxidase activation in inflammatory cells, as well as activation of nuclear factor-kappa B, cyclooxygenase-2, and inducible nitric oxide synthase. Oral administration of COS also reduced serum levels of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-6). Moreover, it prolonged survival time in mice. These data suggest that COS have anti-inflammatory effects in an experimental model of IBD, and could be new functional foods for IBD patients.

Modified administration schedule of docetaxel, cisplatin, and fluorouracil for advanced or recurrent gastric cancer with gastrointestinal stenosis.

PURPOSE: To evaluate the safety and efficacy of a modified administration schedule of docetaxel, cisplatin, and fluorouracil (mDCF)in patients with advanced gastric cancer with gastrointestinal stenosis in clinical practice. METHODS: In the chemotherapy-naïve patients who had metastatic or recurrent histologically confirmed gastric cancer, docetaxel(40mg/m2), levofolinate(200mg/m / / / 2), fluorouracil(400mg/m2)on day 1, fluorouracil 1,000 mg/m2d-2 days intravenous continuous in fusion beginning on day 1, and cisplatin(40mg/m2)on day 3 was administered every 2 weeks. RESULTS: Six patients received mDCF therapy. In 5 patients with measurable disease, the overall response rate was 86%. Median progression-free survival was 310 days and median overall survival was 599 days. Symptom improvement after the first cycle of mDCF was obtained in all patients. Grade 3 or 4 leukopenia and neutropenia were observed in 2(33%)and 6(100%)patients, respectively. There were no treatment-related deaths. CONCLUSION: mDCF seems to be active against metastatic and recurrent gastric cancer with gastrointestinal stenosis. Further study is needed to confirm the efficacy and safety of mDCF regimen.

Anti-tumor properties of orally administered glucosamine and N-acetyl-D-glucosamine oligomers in a mouse model.

The current study evaluated the anti-tumor activities of N-acetyl-d-glucosamine oligomer (NACOS) and glucosamine oligomer (COS) after their oral administration in a tumor (colon 26)-bearing mouse model. Compared to the control group, NACOS and COS groups showed significantly suppressed tumor growth, and apparent, marked apoptosis in tumor tissues. Furthermore, serum interleukin-12p70 and interferon-γ levels significantly increased in the NACOS and COS groups compared to the corresponding levels in the control group. Collectively, the results indicate the oral administration of NACOS and COS could enhance innate immunity. Results of experiments in Myd-88 knockout mice revealed that the apparent effects were related to both Myd-88-dependent and Myd-88-independent pathways. The data indicated that oral administration of NACOS and COS produced anti-tumor effects through the induction of apoptosis and stimulation of the immune system, which suggests that NACOS and COS are candidate anti-tumor functional foods.

Depolymerization of sulfated polysaccharides under hydrothermal conditions.

Fucoidan and chondroitin sulfate, which are well known sulfated polysaccharides, were depolymerized under hydrothermal conditions (120-180°C, 5-60min) as a method for the preparation of sulfated polysaccharides with controlled molecular weights. Fucoidan was easily depolymerized, and the change of the molecular weight values depended on the reaction temperature and time. The degree of sulfation and IR spectra of the depolymerized fucoidan did not change compared with those of untreated fucoidan at reaction temperatures below 140°C. However, fucoidan was partially degraded during depolymerization above 160°C. Nearly the same depolymerization was observed for chondroitin sulfate. These results indicate that hydrothermal treatment is applicable for the depolymerization of sulfated polysaccharides, and that low molecular weight products without desulfation and deformation of the initial glycan structures can be obtained under mild hydrothermal conditions.

Novel carbapenem antibiotics for parenteral and oral applications: in vitro and in vivo activities of 2-aryl carbapenems and their pharmacokinetics in laboratory animals.

SM-295291 and SM-369926 are new parenteral 2-aryl carbapenems with strong activity against major causative pathogens of community-acquired infections such as methicillin-susceptible Staphylococcus aureus, Streptococcus pneumoniae (including penicillin-resistant strains), Streptococcus pyogenes, Enterococcus faecalis, Klebsiella pneumoniae, Moraxella catarrhalis, Haemophilus influenzae (including ß-lactamase-negative ampicillin-resistant strains), and Neisseria gonorrhoeae (including ciprofloxacin-resistant strains), with MIC(90)s of ≤ 1 µg/ml. Unlike tebipenem (MIC(50), 8 µg/ml), SM-295291 and SM-369926 had no activity against hospital pathogens such as Pseudomonas aeruginosa (MIC(50), ≥ 128 µg/ml). The bactericidal activities of SM-295291 and SM-369926 against penicillin-resistant S. pneumoniae and ß-lactamase-negative ampicillin-resistant H. influenzae were equal or superior to that of tebipenem and greater than that of cefditoren. The therapeutic efficacies of intravenous administrations of SM-295291 and SM-369926 against experimentally induced infections in mice caused by penicillin-resistant S. pneumoniae and ß-lactamase-negative ampicillin-resistant H. influenzae were equal or superior to that of tebipenem and greater than that of cefditoren, respectively, reflecting their in vitro activities. SM-295291 and SM-369926 showed intravenous pharmacokinetics similar to those of meropenem in terms of half-life in monkeys (0.4 h) and were stable against human dehydropeptidase I. SM-368589 and SM-375769, which are medoxomil esters of SM-295291 and SM-369926, respectively, showed good oral bioavailability in rats, dogs, and monkeys (4.2 to 62.3%). Thus, 2-aryl carbapenems are promising candidates that show an ideal broad spectrum for the treatment of community-acquired infections, including infections caused by penicillin-resistant S. pneumoniae and ß-lactamase-negative ampicillin-resistant H. influenzae, have low selective pressure on antipseudomonal carbapenem-resistant nosocomial pathogens, and allow parenteral, oral, and switch therapies.

Unimolecular and bimolecular binding system for the prediction of CYP2D6-mediated metabolism.

We have developed a template system for the prediction of CYP2D6-mediated metabolism of compounds. The system is composed of two types of two-dimensional templates (templates A and B), which were generated from mutually occupied areas of typical CYP2D6 substrates. The areas of templates are expressed as hexagonal blocks for application to the two-dimensional structures of chemicals. Experiments with 93 reactions with 69 typical substrates indicated the necessity for two similar but distinct shapes for template A (A1 and A2) for optimal placement. A frequently occupied area for substrates in template A1 was defined as a trigger area in which to capture a substrate for initiation of metabolism. Another frequently occupied area was found near the site of metabolism in template B. Both frequently occupied areas are linked to a pinching area. Occupancy of substrates on two template areas is suggested to be essential for the metabolism of CYPD6 substrates. In cases of CYP2D6 substrates without simultaneous occupancy of both areas, bimolecular placement, in which two molecules are placed coordinately, is proposed. Metabolism of small molecules, including naphthalene and quinoline, became explainable with the use of this idea. Validation of this template system with the use of both good and poor CYP2D6 substrates indicated clear advantages of the present system as a tool for drug modification, in addition to enabling highly accurate estimation of the site of metabolism.

A new methodology for simultaneous quantification of total-Aß, Aßx-38, Aßx-40, and Aßx-42 by column-switching LC/MS/MS.

This article details the development of a novel method that overcomes the drawbacks of sandwich ELISA (sELISA) and allows reliable evaluation of simultaneous quantification of the amyloid (Aß)-peptides, total-Aß, Aßx-38, Aßx-40, and Aßx-42, in rat brain by optimized sample purification and column-switching liquid chromatographic-tandem mass spectrometry (LC/MS/MS). This method provides accurate analyses of total-Aß, Aßx-38, Aßx-40, and Aßx-42 with a linear calibration range between 0.05 and 45 ng/mL. Verification for accuracy and precision of biological samples were determined by a standard addition and recovery test, spiked with synthetic Aß1-38, Aß1-40, and Aß1-42 into the rat brain homogenate. This method showed <20% relative error and relative standard deviation, indicating high reproducibility and reliability. The brain concentrations of total-Aß, Aßx-38, Aßx-40, and Aßx-42 after oral administration of flurbiprofen in rats were measured by this method. Aßx-42 concentrations (4.57 ± 0.69 ng/g) in rats administered flurbiprofen were lower than those in untreated rats (6.48 ± 0.93 ng/g). This was consistent with several reports demonstrating that NSAIDs reduced the generation of Aß. We report here a method that allows not only the quantification of specific molecular species of Aß but also simultaneous quantification of total-Aß, Aßx-38, Aßx-40, and Aßx-42, thus overcoming the drawbacks of sELISA.

Localization of mature neprilysin in lipid rafts.

Alzheimer's disease (AD) is characterized by senile plaques caused by amyloid-ß peptide (Aß) accumulation. It has been reported that Aß generation and accumulation occur in membrane microdomains, called lipid rafts, which are enriched in cholesterol and glycosphingolipids. Moreover, the ablation of cholesterol metabolism has been implicated in AD. Neprilysin (NEP), a neutral endopeptidase, is one of the major Aß-degrading enzymes in the brain. Activation of NEP is a possible therapeutic target. However, it remains unknown whether the activity of NEP is regulated by its association with lipid rafts. Here we show that only the mature form of NEP, which has been glycosylated in the Golgi, exists in lipid rafts, where it is directly associated with phosphatidylserine. Moreover, the localization of NEP in lipid rafts is enhanced by its dimerization, as shown using the NEP E403C homodimerization mutant. However, the protease activities of the mature form of NEP, as assessed by in vitro peptide hydrolysis, did not differ between lipid rafts and nonlipid rafts. We conclude that cholesterol and other lipids regulate the localization of mature NEP to lipid rafts, where the substrate Aß accumulates but does not modulate the protease activity of NEP.

Crystal structures of protein glutaminase and its pro forms converted into enzyme-substrate complex.

Protein glutaminase, which converts a protein glutamine residue to a glutamate residue, is expected to be useful as a new food-processing enzyme. The crystal structures of the mature and pro forms of the enzyme were refined at 1.15 and 1.73 Å resolution, respectively. The overall structure of the mature enzyme has a weak homology to the core domain of human transglutaminase-2. The catalytic triad (Cys-His-Asp) common to transglutaminases and cysteine proteases is located in the bottom of the active site pocket. The structure of the recombinant pro form shows that a short loop between S2 and S3 in the proregion covers and interacts with the active site of the mature region, mimicking the protein substrate of the enzyme. Ala-47 is located just above the pocket of the active site. Two mutant structures (A47Q-1 and A47Q-2) refined at 1.5 Å resolution were found to correspond to the enzyme-substrate complex and an S-acyl intermediate. Based on these structures, the catalytic mechanism of protein glutaminase is proposed.