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A pregnant woman with severe aplastic anemia was managed using biweekly red blood cell transfusion and oral eltrombopag olamine administration during pregnancy. She was diagnosed with preeclampsia at 35 weeks of gestation. The severity of aplastic anemia is very important for predicting the course of pregnancy. The frequency of preeclampsia in pregnant women with aplastic anemia may be higher than that in general pregnancies. Attention is paid to severe anemia in the early trimesters. Eltrombopag can be used in pregnancy. [ABSTRACT FROM AUTHOR]

In this work, 3D printable gel polymer electrolytes (GPEs) based on N,N‐dimethylacrylamide (DMAAm) and polyvinylidene fluoride (PVDF) in lithium chloride containing ethylene glycol solution are synthesized and their physicochemical properties are investigated. 3D printing is carried out with a customized stereolithography type 3D gel printer named "Soft and Wet Intelligent Matter‐Easy Realizer" and free forming GPE samples having variable shapes and sizes are obtained. Printed PVDF/PDMAAm‐based GPEs exhibit tunable mechanical properties and favorable thermal stability. Electrochemical proprieties of the printed GPEs are carried out via impedance spectroscopy in the temperature range of 25–90 °C by varying PVDF content. Ionic conductivity as high as 6.5 × 10−4 S cm−1 is achieved at room temperature for GPE containing low PVDF content (5 wt%) and conductivity of the GPEs is increased as temperature rises. [ABSTRACT FROM AUTHOR]

Nonalcoholic steatohepatitis (NASH) is a common liver disease that occurs in both alcoholics and nonalcoholics. Oxidative stress is a possible causative factor for liver diseases including NASH. Gut microorganisms, especially lactic acid bacteria, can produce unique fatty acids, including hydroxy, oxo, conjugated, and partially saturated fatty acids. The oxo fatty acid 10‐oxo‐11(E)‐octadecenoic acid (KetoC) provides potent cytoprotective effects against oxidative stress through activation of Nrf2‐ARE pathway. The aim of this study was to explore the preventive and therapeutic effects of gut microbial fatty acid metabolites in a NASH mouse model. The mice were divided into 3 experimental groups and fed as follows: (1) high‐fat diet (HFD) (2) HFD mixed with 0.1% KetoA (10‐oxo‐12(Z)‐octadecenoic acid), and (3) HFD mixed with 0.1% KetoC. After 3 weeks of feeding, plasma parameters, liver histology, and mRNA expression of multiple genes were assessed. There was hardly any difference in fat accumulation in the histological study; however, no ballooning occurred in 2/5 mice of KetoC group. Bridging fibrosis was not observed in the KetoA group, although KetoA administration did not significantly suppress fibrosis score (p = 0.10). In addition, KetoC increased the expression level of HDL related genes and HDL cholesterol levels in the plasma. These results indicated that KetoA and KetoC may partly affect the progression of NASH in mice models. [ABSTRACT FROM AUTHOR]

Dietary intake of ω3 polyunsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid is beneficial for health control. We recently identified 17,18‐epoxyeicosatetraenoic acid (17,18‐EpETE) as a lipid metabolite endogenously generated from eicosapentaenoic acid that exhibits potent anti‐allergic and anti‐inflammatory properties. However, chemically synthesized 17,18‐EpETE is enantiomeric due to its epoxy group—17(S),18(R)‐EpETE and 17(R),18(S)‐EpETE. In this study, we demonstrated stereoselective differences of 17(S),18(R)‐EpETE and 17(R),18(S)‐EpETE in amelioration of skin contact hypersensitivity and found that anti‐inflammatory activity was detected in 17(S),18(R)‐EpETE, but not in 17(R),18(S)‐EpETE. In addition, we found that cytochrome P450 BM‐3 derived from Bacillus megaterium stereoselectively converts EPA into 17(S),18(R)‐EpETE, which effectively inhibited the development of skin contact hypersensitivity by inhibiting neutrophil migration in a G protein‐coupled receptor 40‐dependent manner. These results suggest the new availability of a bacterial enzyme to produce a beneficial lipid mediator, 17(S),18(R)‐EpETE, in a stereoselective manner. Our findings highlight that bacterial enzymatic conversion of fatty acid is a promising strategy for mass production of bioactive lipid metabolites. [ABSTRACT FROM AUTHOR]

Background: The bioactive metabolite KetoC, generated by intestinal bacteria, exerts various beneficial effects. Nevertheless, its function in the pathogenesis of periodontitis remains unclear. Here, we investigated the effect of KetoC in a mouse model of periodontitis and explored the underlying mechanism.Methods: Thirty-one 8-week-old male C57BL/6N mice were randomly divided into four groups (non-ligation, non-ligation + KetoC, ligation + Porphyromonas gingivalis, and ligation + P. gingivalis + KetoC) (n = 7/8 mice/group) and given a daily oral gavage of KetoC (15 mg/mL) or vehicle for 2 weeks. To induce periodontitis, a 5-0 silk ligature was placed on the maxillary left second molar on day 7, and P. gingivalis W83 (109 colony-forming unit [CFU]) was administered orally every 3 days. On day 14, all mice were euthanized. Alveolar bone destruction was determined from the level of the cemento-enamel junction to the alveolar bone crest. Moreover, bone loss level was confirmed from gingival tissue sections stained with hematoxylin and eosin. The presence of P. gingivalis was quantified using real-time polymerase chain reaction. In vitro, the bacteriostatic and bactericidal effects of KetoC were assessed by analyzing its suppressive activity on the proliferation of P. gingivalis and using a live/dead bacterial staining kit, respectively. A double-bond-deficient metabolite (KetoB) was then used to investigate the importance of double-bond structure in the antimicrobial activity of KetoC on P. gingivalis.Results: In vivo, KetoC attenuated alveolar bone destruction and suppressed P. gingivalis in the periodontitis group. In vitro, KetoC (but not KetoB) downregulated the proliferation and viability of P. gingivalis in a dose-dependent manner.Conclusions: KetoC reduced alveolar bone destruction in a periodontitis model via its antimicrobial function. Therefore, this bioactive metabolite may be valuable in clinical applications to support periodontal therapy. [ABSTRACT FROM AUTHOR]

The oleaginous fungus Mortierella alpina is known to accumulate eicosapentaenoic acid (EPA) only when cultivated at a low temperature (below 15°C). Here, we investigated EPA production at an ordinary temperature (28°C) by expressing the Saprolegnia diclina D17 desaturase gene (sdd17m) in M. alpina ST1358, an ω3-desaturation activity-defective mutant derived from M. alpina 1S-4. Expression of the exogenous gene was confirmed by EPA accumulation in transformants at both 28 and 12°C. The EPA content in total lipids produced by transformants was over 20% at 28°C. Bench-scale fermentation with a 5-L jar fermentor showed that EPA content reached 26.4% of total fatty acids, and EPA production reached 1.8 g/L. This is the first study to report the accumulation of EPA in M. alpina at an ordinary temperature, and provide a platform technology for the industrial production of EPA using M. alpina as a promising source for EPA. [ABSTRACT FROM AUTHOR]

Hydroxy and oxo fatty acids were recently found to be produced as intermediates during gut microbial fatty acid metabolism. Lactobacillus plantarum produces these fatty acids from unsaturated fatty acids such as linoleic acid. In this study, we investigated the effects of these gut microbial fatty acid metabolites on the lipogenesis in liver cells. We screened their effect on sterol regulatory element binding protein-1c (SREBP-1c) expression in HepG2 cells treated with a synthetic liver X receptor α (LXRα) agonist (T0901317). The results showed that 10-hydroxy-12( Z)-octadecenoic acid (18:1) (HYA), 10-hydroxy-6( Z),12( Z)-octadecadienoic acid (18:2) (γHYA), 10-oxo-12( Z)-18:1 (KetoA), and 10-oxo-6( Z),12( Z)-18:2 (γKetoA) significantly decreased SREBP- 1c mRNA expression induced by T0901317. These fatty acids also downregulated the mRNA expression of lipogenic genes by suppressing LXRα activity and inhibiting SREBP-1 maturation. Oral administration of KetoA, which effectively reduced triacylglycerol accumulation and acetyl-CoA carboxylase 2 ( ACC2) expression in HepG2 cells, for 2 weeks significantly decreased Srebp- 1c, Scd- 1, and Acc2 expression in the liver of mice fed a high-sucrose diet. Our findings suggest that the hypolipidemic effect of the fatty acid metabolites produced by L. plantarum can be exploited in the treatment of cardiovascular diseases or dyslipidemia. [ABSTRACT FROM AUTHOR]

Recently, a novel gut-bacterial fatty acid metabolism, saturation of polyunsaturated fatty acid, that modifies fatty acid composition of the host and is expected to improve our health by altering lipid metabolism related to the onset of metabolic syndrome, was discovered in Lactobacillus plantarum AKU 1009a. Enzymes constituting the pathway catalyze sequential reactions of free fatty acids without CoA or acyl carrier protein. Among these enzymes, CLA- ER was identified as an enone reductase that can saturate the C=C bond in the 10-oxo- trans-11-octadecenoic acid (KetoB) to produce 10-oxo-octadecanoic acid (KetoC). This enzyme is the sole member of the NADH oxidase/flavin reductase family that has been identified to exert an enone reduction activity. Here, we report both the structure of holo CLA- ER with cofactor FMN and the KetoC-bound structure, which elucidate the structural basis of enone group recognition of free fatty acids and provide the unique catalytic mechanism as an enone reductase in the NADH oxidase/flavin reductase family. A 'cap' structure of CLA- ER underwent a large conformational change upon KetoC binding. The resulting binding site adopts a sandglass shape and is positively charged at one side, which is suitable to recognize a fatty acid molecule with enone group. Based on the crystal structures and enzymatic activities of several mutants, we identified C51, F126 and Y101 as the critical residues for the reaction and proposed an alternative electron transfer pathway of CLA- ER. These findings expand our understanding of the complexity of fatty acid metabolism. Database The atomic coordinates have been deposited in the Protein Data Bank (PDB), (PDB ID , ) [ABSTRACT FROM AUTHOR]

A novel enzymatic production system of optically pure β-hydroxy α-amino acids was developed. Two enzymes were used for the system: an N-succinyl L-amino acid β-hydroxylase (SadA) belonging to the iron(II)/α-ketoglutarate-dependent dioxygenase superfamily and an N-succinyl L-amino acid desuccinylase (LasA). The genes encoding the two enzymes are part of a gene set responsible for the biosynthesis of peptidyl compounds found in the Burkholderia ambifaria AMMD genome. SadA stereoselectively hydroxylated several N-succinyl aliphatic L-amino acids and produced N-succinyl β-hydroxy L-amino acids, such as N-succinyl- L-β-hydroxyvaline, N-succinyl- L-threonine, (2 S,3 R)- N-succinyl- L-β-hydroxyisoleucine, and N-succinyl- L- threo-β-hydroxyleucine. LasA catalyzed the desuccinylation of various N-succinyl- L-amino acids. Surprisingly, LasA is the first amide bond-forming enzyme belonging to the amidohydrolase superfamily, and has succinylation activity towards the amino group of L-leucine. By combining SadA and LasA in a preparative scale production using N-succinyl- L-leucine as substrate, 2.3 mmol of L- threo-β-hydroxyleucine were successfully produced with 93% conversion and over 99% of diastereomeric excess. Consequently, the new production system described in this study has advantages in optical purity and reaction efficiency for application in the mass production of several β-hydroxy α-amino acids. [ABSTRACT FROM AUTHOR]

Thirty-four strains of docosahexaenoic acid (DHA)-producing microorganisms were newly isolated from brackish areas in Japan. These strains showing various compositions of fatty acids. Especially, the fatty acids produced by one of the strains, named D31, had a high DHA content (over 60 % of the total fatty acids) and the simple fatty acid composition (16:0, 18:0, 18:1 and DHA without any other polyunsaturated acids). Although most oleaginous microorganisms accumulate DHA as triacylglycerol, the strain D31 accumulated DHA mainly as a polar lipid (79.4 % of total DHA), especially as phosphatidylcholine (71.4 % of polar DHA). This strain D31 was identified as a related species of Crypthecodinium cohnii on the basis of phylogenetic analysis. Crypthecodinium sp. D31 showed high DHA productivity when cultivated in a medium containing glycerol as the carbon source and a mixture of yeast extract and polypeptone as the nitrogen sources, with a salinity that was equivalent to 50 % of that of seawater and a pH in the acidic range (

Through the screening of about 300 strains of lactic acid bacteria, Pediococcus sp. AKU 1080 was selected as a strain with the ability to hydrate linoleic acid ( cis-9, cis-12-octadecadienoic acid) to three hydroxy fatty acids, i.e., 10-hydroxy- cis-12-octadecenoic acid, 13-hydroxy- cis-9-octadecenoic acid, and 10,13-dihydroxyoctadecanoic acid. The strain hydrated one of two cis double bonds at Δ9 and Δ12 positions to produce 10-hydroxy- cis-12-octadecenoic acid and 13-hydroxy- cis-9-octadecenoic acid, respectively, then further hydrated these two mono-hydroxy fatty acids to 10,13-dihydroxyoctadecanoic acid. The growing cells of this strain were applied to the production of 13-hydroxy- cis-9-octadecenoic acid, that is potential as polymer substrates and functional foods but its specific and efficient production was not established. Under the optimum conditions, 2.3 mg/mL of 13-hydroxy- cis-9-octadecenoic acid was produced from 12.3 mg/mL of linoleic acid with 0.04 mg/mL 10-hydroxy- cis-12-octadecenoic acid (HYA) and 0.05 mg/mL 10,13-dihydroxyoctadecanoic acid in the cultivation medium. Specific production of 13-hydroxy- cis-9-octadecenoic acid was attained using cell-free extracts of the strain as the catalyst. Under the optimum conditions, 0.4 mg/mL of 13-hydroxy- cis-9-octadecenoic acid was produced from 2.0 mg/mL of linoleic acid without HYA and 10,13-dihydroxyoctadecanoic acid. Practical applications: Hydroxy fatty acids are useful as starting materials for industrial chemicals, functional foods, and pharmaceuticals. Regioselective introduction of hydroxyl group to unsaturated fatty acids by microorganisms was applied to hydroxy fatty acid production. Especially, specific production of 13-hydroxy- cis-9-octadecenoic acid, which is useful for the production of 13-oxo-fatty acids with anti-obesity activity, was established in this work. [ABSTRACT FROM AUTHOR]

Microorganisms are promising as producers of various polyunsaturated fatty acids (PUFAs) and as catalysts transforming them into unique molecular species beyond common PUFAs. This article describes PUFA production through chemical mutant- and molecular-breeding of an oleaginous filamentous fungus Mortierella alpina 1S-4 and PUFA transformation by anaerobic bacteria. M. alpina 1S-4 and its mutants and transformants produce oils containing not only common n − 6 and n − 3 PUFAs but also rare PUFAs. Unique PUFA-transforming activities were found in anaerobic bacteria. They isomerized PUFA to conjugated fatty acids and further transformed to partially saturated fatty acids with hydroxyl fatty acids as intermediates. The functions of these unique PUFAs have been attracting much attention for improving our health and for developing new chemical materials. [ABSTRACT FROM AUTHOR]

Background: In cases of thoracic esophageal cancer, multidirectional lymphatic flow from the tumor means that lymph node metastasis can occur in an area extending from the neck to the abdomen. To validate a method for limiting the performance of three-field lymphadenectomy only to patients who need it, we carried out a prospective study in which superparamagnetic iron oxide (SPIO)-enhanced lymphatic mapping was used to determine whether to perform neck lymph node dissection in patients with submucosal thoracic esophageal cancer. Methods: A total of 22 patients with clinically submucosal thoracic squamous cell esophageal cancer, without neck lymph node metastasis, were enrolled. SPIO was endoscopically injected into the peritumoral submucosal layer, after which its appearance in lymph nodes in the neck was evaluated using magnetic resonance imaging (MRI). Neck lymph nodes were then dissected based on the SPIO-enhanced MRI lymphatic mapping. Results: Influx of SPIO into lymph nodes was detected in 21 patients (95% detection rate). SPIO flowed to the neck in 8 (36%) patients. Influx of SPIO into neck lymph nodes was unilateral in five patients and bilateral in three patients, and the lymph nodes were dissected accordingly. A cancer-involved node was identified in two of those patients. In 14 patients, we did not dissect neck nodes. Patients were followed up for 6 to 47 months. The neck lymph node recurrence rate was zero, and the overall recurrence rate was 5%. Conclusions: SPIO-enhanced lymphatic mapping may be useful for estimating the need for three-field lymphadenectomy with neck dissection. [ABSTRACT FROM AUTHOR]

Stress hyperglycemia refers to the transient hyperglycemia seen during illness and is usually restricted to patients without previous evidence of diabetes. The influence of genetics on surgery-induced hyperglycemia remains only partially understood. The study participants were Japanese patients treated for thoracic esophageal cancer with curative esophagectomy at Akita University Hospital between 2003 and 2007. We determined the associations between esophagectomy-induced stress hyperglycemia (≥30 mg/dl increases in blood glucose during surgery) and genetic polymorphisms for C-reactive protein (CRP), tumor necrosis factor (TNF)-α, -β, interferon-γ, transforming growth factor-β1, interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-6 receptors, IL-10, IL-12β, adiponectin, and peroxisome proliferator-activated receptor-γ. In 28 (46%) patients, blood glucose levels increased more than 30 mg/dl during surgery. Among the genetic polymorphisms tested, CRP − 717C> T was significantly associated with stress hyperglycemia during esophagectomy. Multivariate logistic regression revealed that patients with the CRP − 717T/T genotype had a significantly greater risk of developing surgery-induced hyperglycemia than those with the CRP − 717C/T genotype. Stress hyperglycemia was also significantly associated with postoperative infectious complications and duration of intensive care unit stay. It is suggested that CRP − 717 C> T genetic polymorphism may be a predictive factor for stress hyperglycemia in patients receiving esophagectomy for thoracic esophageal cancer. [ABSTRACT FROM AUTHOR]

Following esophagectomy, tracheobronchial lesions (TBLs) can occur as a result of ischemia caused by extensive dissection around the tracheobronchus. In this study we assessed the causes and clinical features of these complications, paying particular attention to lymph node (LN)-related factors. Between January 2000 and March 2007, 305 consecutive patients underwent subtotal esophagectomy using a transthoracic approach with LN dissection for thoracic esophageal cancer. TBLs, including erosions, ulcers, and fistulae, without traumatic injury during the operation, were detected during bronchoscopic examinations performed twice daily after the operation. The correlation between TBLs and tumor or surgical factors were analyzed. TBLs were observed in 14 patients, accounting for an overall incidence of 5%; these included 6 fistulae, 5 ulcers, and 3 erosions. Cases with TBLs significantly more often involved three-field LN dissections (3FLD) than those without TBLs. Six (43%) patients with TBLs had more than four metastatic lymph nodes, while 9 (64%) had cervical and upper-mediastinal LN metastasis ( p = 0.034 and 0.041, respectively). More than 60 LNs were dissected from 10 (71%) patients with TBLs ( p = 0.021), and logistic regression analysis revealed that dissection of more than 60 lymph nodes and 3FLD were independent predictors of TBLs. Esophageal cancer patients requiring extensive LN dissection of more than 60 nodes and/or 3FLD have an increased risk of developing a TBL during their postoperative course. [ABSTRACT FROM AUTHOR]

Three-field lymph node dissection for thoracic esophageal cancer is associated with high morbidity and reduced quality of life after surgery. Consequently, minimized lymphadenectomy would be desirable, if appropriate. In the present study, we retrospectively analyzed the status of involved nodes and the direction of metastatic lymphatic flow from tumors into involved nodes to determine whether submucosal squamous cell esophageal cancers are potential candidates for minimized lymphadenectomy. We enrolled 199 patients who received esophagectomy with extensive lymph node dissection between 1989 and 2005 and retrospectively analyzed their prognoses, distribution of solitary metastatic lymph nodes, and the direction of metastatic lymphatic flow from the tumor, taking into consideration tumor location and depth. Of these patients with submucosal cancers, 83% had 1 or 2 involved nodes, and their esophageal cancer-specific 5-year survival rate was 66%. Solitary lymph node metastasis did not occur in neck lymph nodes in lower thoracic submucosal esophageal cancers, and the direction of metastatic lymphatic flow from the tumor was almost always in one direction. By contrast, T2–4 cancers with 2–4 involved nodes had bidirectional metastatic lymphatic flow from the tumor. There was a difference in the status of lymph node metastasis and the direction of metastatic lymphatic flow from tumors into involved nodes between submucosal and T2–4 thoracic squamous cell esophageal cancers. This analysis may be useful for developing an approach to minimized lymphadenectomy for thoracic esophageal cancers. [ABSTRACT FROM AUTHOR]

Heat shock protein (Hsp) 90 is a key regulator of a variety of oncogene products and cell-signaling molecules, and the therapeutic benefit of its inhibition in combination with radiation or chemotherapy has been investigated. In addition, hyperthermia has been used for many years to treat various malignant tumors. We previously described a system in which hyperthermia was induced using thermosensitive ferromagnetic particles (FMP) with a Curie temperature (Tc = 43˚C) low enough to mediate automatic temperature control, and demonstrated its antitumor effect in a mouse melanoma model. In the present study, we examined the antitumor effects of combining a Hsp90 inhibitor (geldanamycin; GA) with FMP-mediated hyperthermia. In cultured B16 melanoma cells, GA exerted an antitumor effect by increasing the cells’ susceptibility to hyperthermia and reducing expression of Akt. In an in vivo study, melanoma cells were subcutaneously injected into the backs of C57BL/6 mice. FMP were then injected into the resultant tumors, and the mice were divided into four groups: group I, no treatment (control); group II, one hyperthermia treatment; group III, GA alone; and group IV, GA with hyperthermia. When exposed to a magnetic field, the temperature of tissues containing FMP increased and stabilized at the Tc. In group IV, complete regression of tumors was observed in five of nine mice (56%), whereas no tumor regression was seen in groups I–III. Our findings suggest that inhibition of Hsp90 with hyperthermia increases its antitumor effect. Thus, the combination of FMP-mediated, self-regulating hyperthermia with Hsp90 inhibition has important implications for the treatment of cancer. ( Cancer Sci 2009; 100: 558–564) [ABSTRACT FROM AUTHOR]

The fatty acid desaturation and elongation reactions catalyzed by Trichoderma sp. 1-OH-2-3 were investigated. This strain converted palmitic acid (16:0) mainly to stearic acid (18:0), and further to oleic acid ( c9-18:1), linoleic acid ( c9, c12-18:2), and α-linolenic acid ( c9, c12, c15-18:3) through elongation, and Δ9, Δ12, and Δ15 desaturation reactions, respectively. Palmitoleic acid ( c9-16:1) and cis-9, cis-12-hexadecadienoic acid were also produced from 16:0 by the strain. This strain converted n-tridecanoic acid (13:0) to cis-9-heptadecenoic acid and further to cis-9, cis-12-heptadecadienoic acid through elongation, and Δ9 and Δ12 desaturation reactions, respectively. trans-Vaccenic acid ( t11-18:1) and trans-12-octadecenoic acid ( t12-18:1) were desaturated by the strain through Δ9 desaturation. The products derived from t11-18:1 were identified as the conjugated linoleic acids (CLAs) of cis-9, trans-11-octadecadienoic acid and trans-9, trans-11-octadecadienoic acid. The product derived from t12-18:1 was identified as cis-9, trans-12-octadecadienoic acid. cis-6, cis-9-Octadecadienoic acid was desaturated to cis-6, cis-9, cis-12-octadecatrienoic acid by this strain through Δ12 desaturation. The broad substrate specificity of the elongation, and Δ9 and Δ12 desaturation reactions of the strain is useful for fatty acid biotransformation. [ABSTRACT FROM AUTHOR]

Identification of reliable markers of chemo- and radiosensitivity and the key molecules that enhance the susceptibility of squamous esophageal cancer cells to anticancer treatments would be highly desirable. To test whether regenerating gene ( REG) I expression enhances chemo- and radiosensitivity in esophageal squamous cell carcinoma cells, we used MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assays to compare the chemo- and radiosensitivities of untransfected TE-5 and TE-9 cells with those of cells stably transfected with REG Iα and Iβ. We then used flow cytometry to determine whether REG I expression alters cell cycle progression. No REG I mRNA or protein were detected in untransfected TE-5 and TE-9 cells. Transfection with REG Iα and Iβ led to strong expression of both REG I mRNA and protein in TE-5 and TE-9 cells, which in turn led to significant increases in both chemo- and radiosensitivity. Cell cycle progression was unaffected by REG I expression. REG I thus appears to enhance the chemo- and radiosensitivity of squamous esophageal cancer cells, which suggests that it may be a useful target for improved and more individualized treatments for patients with esophageal squamous cell carcinoma. ( Cancer Sci 2008; 99: 2491–2495) [ABSTRACT FROM AUTHOR]

Hyperthermia has been used for many years to treat a variety of malignant tumors. The Curie temperature (Tc) is a transition point at which magnetic materials lose their magnetic properties, causing a cessation of current and thus heat production. The Tc enables automatic temperature control throughout a tumor as a result of the self-regulating nature of the thermosensitive material. We have developed a method of magnetically-induced hyperthermia using thermosensitive ferromagnetic particles (FMPs) with low Tc (43°C), enough to mediate automatic temperature control. B16 melanoma cells were subcutaneously injected into the backs of C57BL/6 mice, after which tumors were allowed to grow to 5 mm in diameter. FMPs were then injected into the tumors, and the mice were divided into three groups: group I (no hyperthermia, control); group II (one hyperthermia treatment); and group III (hyperthermia twice a week for 4 weeks). When exposed to a magnetic field, the FMPs showed a sharp rise in heat production, reaching the Tc in tissue within 7 min, after which the tissue temperature stabilized at approximately the Tc. In groups I and II, all mice died within 30–45 days. In group III, however, 6 of 10 mice remained alive 120 days after beginning treatment. Our findings suggest that repeated treatment with magnetically-induced self-regulating hyperthermia, mediated by FMPs with a low Tc, is an effective means of suppressing melanoma growth. A key advantage of this hyperthermia system is that it is minimally invasive, requiring only a single injection for repeated treatments with automatic temperature control. ( Cancer Sci 2008; 99: 805–809) [ABSTRACT FROM AUTHOR]

Denervated stomach used as an esophageal substitute after esophagectomy often retains or spontaneously recovers acid production. The aims of the present study were to assess the relationship between esophageal acid exposure or gastric acidity and reflux-related symptoms after esophagectomy, and to assess the ability of proton pump inhibitors (PPIs) to relieve gastroesophageal reflux-related symptoms. Forty-four patients underwent esophageal and gastric 24-h pH monitoring early after esophagectomy with gastric reconstruction. Initially, patients with both gastric acidity and reflux symptoms were treated with PPIs (Treatment group), then all patients with gastric acidity, whether symptomatic or not, were treated (Prevention group). Reflux-related symptoms were correlated with esophageal acid exposure and postoperative gastric acidity. Gastric acidity was then correlated with serum anti- Helicobacter pylori immunoglobulin G (IgG) titers and preoperative endoscopic findings Sixteen patients (36.4%) reporting reflux and showing gastric acid production were treated with PPIs, which provided relief to 13 (81.3%). Although symptoms did not correlate with the esophageal acid exposure, postoperative gastric acidity was significantly greater among patients who were symptomatic than among those who were not. Overall, acid production was lower in older patients (> 64 years of age), although older patients who were H. pylori-negative and without chronic atrophic gastritis also showed high levels of gastric acidity. Proton pump inhibitors should be administered prophylactically early after esophagectomy to relieve and prevent reflux-related symptoms. Candidates for preventive therapy include those less than 64 years of age or older patients who are H. pylori-negative and without chronic atrophic gastritis. [ABSTRACT FROM AUTHOR]

The aim of the present study was to assess the outcome of treatment for patients with recurrent mid- and lower-thoracic esophageal cancers in whom recurrence was localized to the lymph nodes of the neck, and to determine the best strategy for further treatment. Between 1989 and 2001, 270 patients with mid- and lower-thoracic esophageal cancer underwent curative esophagectomy; 90 of those patients had a cancer recurrence. Our focus was on lymph node recurrence, especially when the recurrent cancers were localized to the lymph nodes in the neck. The outcomes of those patients and the efficacy of the strategies used to treat the recurrent cancers were determined. In 43 patients (48%), recurrent cancer initially appeared in the lymph nodes. Among the 43 patients, 15 (35%) had localized neck recurrence. The time between tumor recurrence and death among the 15 patients with localized neck recurrence was significantly longer than among the 28 patients with other recurrence patterns. In addition, 15 patients underwent lymph node resection, while 28 patients were treated non-surgically. The time between tumor recurrence and death was significantly longer in patients treated surgically. Of the 15 patients in whom recurrence affected the neck lymph nodes only, 10 (67%) were treated surgically; their 2-year survival rate after recurrence was 45%. The outcomes of recurrent esophageal cancers localized to the lymph nodes of the neck were better than those seen with other recurrence patterns, and salvage resection followed by chemoradiation therapy would seem to be indicated for those patients. [ABSTRACT FROM AUTHOR]

A mixture of oil/ethanol (1∶3, w/w) was shaken at 30°C with 4% immobilized Candida antarctica lipase by weight of the reaction mixture. The reaction regiospecifically converted FA at the 1- and 3-positions to FA ethyl esters, and the lipase acted on C14−C24 FA to a similar degree. The content of 2-MAG reached a maximum after 4 h; the content was 28–29 mol% based on the total amount of FA in the reaction mixture at 59–69% ethanolysis. Only 2-MAG were present in the reaction mixture during the first 4 h, and 1(3)-MAG were detected after 7 h. After removal of ethanol from the 4-h reaction mixture by evaporation, 2-MAG were fractionated by silica gel column chromatography. The contents of FA in the 2-MAG obtained by ethanolysis of several oils coincided well with FA compositions at the 2-position, which was analyzed by Grignard degradation. It was shown that ethanolysis of oil with C. antarctica lipase can be applied to analysis of FA composition at the 2-position in TAG. [ABSTRACT FROM AUTHOR]

The ability to produce CLA from ricinoleic acid is widely distributed in lactic acid bacteria. Washed cells of Lactobacillus plantarum JCM 1551 were selected as a potential catalyst for CLA production from ricinoleic acid. Cells cultivated in medium supplemented with a mixture of α-linolenic acid and linoleic acid showed enhanced CLA productivity. Under optimal reaction conditions, with the free acid form of ricinoleic acid as the substrate and washed cells of L. plantarum as the catalyst, 2.4 mg/mL CLA was produced from 3.4 mg/mL ricinoleic acid in 90 h, with a molar yield with respect to ricinoleic acid of 71%. The CLA produced, which was obtained in the FFA form, consisted of a mixture of two CLA isomers, cis-9, trans-11-octadecadienoic acid (21% of total CLA) and trans-9, trans-11-octadecadienoic acid (79% of total CLA), and accounted for 72% of the total FA obtained. [ABSTRACT FROM AUTHOR]

After screening 14 genera of lactic acid bacteria, Lactobacillus plantarum AKU 1009a was selected as a potential strain for CLA production from linoleic acid. Washed cells of L. plantarum with high levels of CLA production were obtained by cultivation in a nutrient medium with 0.06% (wt/vol) linoleic acid ( cis-9, cis-12-octadecadienoic acid). Under the optimal reaction conditions with the free form of linoleic acid as the substrate, washed cells of L. plantarum produced 40 mg CLA/mL reaction mixture (33% molar yield) from 12% (wt/vol) linoleic acid in 108 h. The resulting CLA was a mixture of two CLA isomers, cis-9, trans-11 (or trans-9, cis-11)-octadecadienoic acid (CLA1, 38% of total CLA) and trans-9, trans-11-octadecadienoic acid (CLA2, 62% of total CLA), and accounted for 50% of the total FA obtained. A higher yield (80% molar yield to linoleic acid) was attained with 2.6% (wt/vol) linoleic acid as the substrate in 96 h, resulting in CLA production of 20 mg/mL reaction mixture [consisting of CLA1 (2%) and CLA2 (98%)] and accounting for 80% of total FA obtained. Most of the CLA produced was associated with the cells ( ca. 380 mg CLA/g dry cells), mainly as FFA. [ABSTRACT FROM AUTHOR]

Aims/Background: Rat liver perfused with an oxygenated buffered solution alone results in degenerative changes even when the perfusion flow is accelerated to give a sufficient oxygen supply. On the other hand, perfusion media supplemented with red blood cells (RBCs) preserve the viability of the liver. The present study was conducted to clarify how RBCs protect the isolated perfused liver. Methods: The liver was perfused with and without RBCs in a perfusate equilibrated with supra-physiological oxygen tension at regulated inflow pressures, and controlled hepatic oxygen consumption. We examined alanine aminotransferase and purine nucleoside phosphorylase activity in the perfusate as specific markers of liver cells injury. Hydrogen peroxide (H2O2) production and morphological changes were determined using cerium electron microscopy. Apoptosis was detected by measuring CPP 32 protease activity and using TdT-mediated dUTP-digoxigenin nick end-labeling. Results: When the liver was perfused with RBC-free buffer, H2O2 production and consequent injury progressing to apoptosis were initiated in the sinusoidal endothelial cells (SECs). After SECs were injured, H2O2 appeared in the hepatocytes. H2O2 production and associated degenerative changes were attenuated both morphologically and enzymatically by the addition of RBCs, a specific xanthine oxidase (XOD) inhibitor and the H2O2 radical scavenger, catalase. Conclusions: In the liver perfused with RBC-free buffer, H2O2 production and consequent injury were initiated in SECs. RBCs attenuate liver injury by scavenging XOD-dependent H2O2. [ABSTRACT FROM AUTHOR]

The Δ5 and Δ12 desaturase (DS)-blocked mutants of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, were obtained. These mutants accumulated 8,11- cis-eicosadienoic acid (20:2n-9). One of the mutants, M226-9, in which Δ5 and Δ12 DS are perfectly blocked, produced 1.68 mg of 20:2n-9 per mL of culture medium (101 mg/g dry mycelia) and no 5,8,11- cis-eicosatrienoic acid (20:3n-9) when grown in a medium containing 4% glucose and 1% yeast extract at 28°C for 2 d and then at 12°C for 12 d. The mycelial lipid comprised 77.4% triacylglycerol (TG) and 9.8% phosphatidylcholine (PC), among others. TG contained 69.0% of the total 20:2n-9, whose percentage in total TG fatty acids was 15.9%. The highest percentage (44.4%) of 20:2n-9 was found in PC. The addition of olive oil to the culture medium enhanced the production of 20:2n-9. [ABSTRACT FROM AUTHOR]

N-Methylhydantoin amidohydrolase, an ATP-dependent amidohydrolase involved in microbial degradation of creatinine, was purified 70-fold to homogeneity, with a 62% overall recovery, and was crystallized from Pseudomonas putida 77. The enzyme has a relative molecular mass of 300 000. It is a tetramer of two identical small subunits (Mr 70 000) and two identical large subunits (Mr 80 000). The enzyme requires ATP for the amidohydrolysis of N-methylhydantoin and vice versa. Mg2+, Mn2+ or Co2+, and K+, NH4+, Rb+ or Cs+, were absolutely required concomitantly for the enzyme activity as divalent and monovalent cations, respectively. The Km and Vmax values for N-methylhydantoin were 32 µM and 9.0 µmol · min-1 · mg protein-1. The hydrolysis of amide compounds and coupled hydrolysis of ATP were observed with hydantoin, DL-5-methylhydantoin, glutarimide and succimide in addition to N-methylhydantoin. 2-Pyrrolidone, 2-oxazolidone, δ-valerolactam, 2,4-thiazolidinedione, 2-imidazolidone, D-5oxoproline methyl ester, DL-5-oxoproline methyl ester, and naturally occurring pyrimidine compounds, i.e. dihydrouracil, dihydrothymine, uracil, and thymine, effectively stimulated ATP hydrolysis by the enzyme without undergoing detectable self-hydrolysis. [ABSTRACT FROM AUTHOR]

β-Ureidopropionase of aerobic bacterial origin was purified to homogeneity from Pseudomonas putida IFO 12996. The enzyme shows a broad substrate specificity. In addition to β-ureidopropionate (Km 3.74 mM, Vmax 4.12 U/mg). γ-ureido-n-butyrate (Km 11.6 mM, V max 19.4 U/mg), and several N-carbamoyl-α-amino acids, such as N-carbamoylglycine (Kmax 0.68 mM, Vmax 9.14x10-2U/mg). N-carbamoyl-L-alanine (Km 1.56 mM, Vmax 1,00 U/mg), N-carbamoyl-L-serine (Km 75.1 mM. Vmax 3.78 U/mg), and N-carbamoyl-DL-α-amino-n-butyrate (Km 2.81 mM. Vmax 1.08 U/mg), are also hydrolyzed. The hydrolysis of N-carbamoyl-α-amino acids is strictly L enantiomer specific. N-Formyl-c-alanine and N-acetyl-L-alanine are also hydrolyzed by the enzyme, but the rate of hydrolysis is lower than the rate for N-carbamoyl-L-alanine. The enzyme requires a divalent metal ion. such as Co2+, Ni2+ or Mn2+, for activity, and is significantly affected by sulfhydryl reagents. The enzyme consists of two polypeptide chains with identical relative molecular mass M1 45000. The broad substrate specificity and metal ion dependence of the enzyme show that the β-ureidopropionase of this aerobic bacterium is quite different from the β-ureidopropionases of mammals and anaerobic bacteria. [ABSTRACT FROM AUTHOR]

N-Carbamoyl-D-amino acid amidohydrolase was purified 119-fold, with 36% overall recovery from a cell-free extract of Comamonas sp. E222c. The purified enzyme was homogeneous as judged by SDS/PAGE. The relative molecular mass of the native enzyme was 120000 and that of the subunit was 40000. The purified enzyme hydrolyzed various N-carbamoyl-D-amino acids to Damino acids, ammonia and carbon dioxide. N-Carbamoyl-D-amino acids having hydrophobic groups served as good substrates for the enzyme. The Km and Vmax values for N-carbamoyl-D-phenylalanine were 19.7 mM and 13.1 units/mg, respectively, and those for N-carbamoyl-D-p-hydroxyphenylglycine were 13.1 mM and 0.56 units/mg, respectively. The enzyme strictly recognized the configuration of the substrate and only the D-enantiomer of the N-carbamoyl amino acid was hydrolyzed. The enzyme activity was not significantly affected by N-carbamoyl-L-amino acids and ammonia. The enzyme was sensitive to thiol reagents and did not require metal ions for its activity. The enzyme did not hydrolyze N-carbamoyl-β-alanine or N-carbamoyl-DL-aspartate suggesting that the enzyme is different from the N-carbamoylamide-hydrolyzing enzymes involved in the pyrimidine degradation pathway. The enzyme did not hydrolyze allantoin and allantoic acid, which are intermediates in purine degradation, N-carbamoylsarcosine and citrulline, suggesting that it is a novel N-carbamoylamide amidohydrolase. [ABSTRACT FROM AUTHOR]

A novel enzyme, arylalkyl acylamidase, which shows a strict specificity for N-acetyl arylalkylamines, but not acetanilide derivatives, was purified from the culture broth of Pseudomonas putida Sc2. The purified enzyme appeared to be homogeneous, as judged by native and SDS/PAGE. The enzyme has a molecular mass of approximately 150 kDa-and consists of four identical subunits. The purified enzyme catalyzed the hydrolysis of N-acetyl-2-phenylethylamine to 2-phenylethylamine and acetic acid at the rate of 6.25 μmol · min-1 · mg-1 at 30°C. It also catalyzed the hydrolysis of various N-acetyl arylalkylamines containing a benzene or indole ring, and acetic acid arylalkyl esters. The enzyme did not hydrolyze acetanilide, N-acetyl aliphatic amines, N-acetyl amino acids, N-acetyl amino sugars or acylthiochotine. The apparent Km for N-acetylbenzylamine, N-acetyl-2-phenyl-ethylamine and N-acetyl-3-phenylpropylamine are 41 mM, 0.31 mM and 1.6 mM, respectively. The purified enzyme was sensitive to thiol reagents such as Ag2SO4, HgCl2 and p-chloromercuribenzoic acid, and its activity was enhanced by divalent metal ions such as Zn2+, Mg2+ and Mn2+. [ABSTRACT FROM AUTHOR]

Imidase, which preferably hydrolyzed cyclic imides to monoamidated dicarboxylates, was purified to homogeneity from a cell-free extract of Blastobacter sp. A17p-4. Cyclic imides are known to be hydrolyzed by mammalian dihydropyrimidinases. However. imidase was quite different from known dihydropyrimidinases in structure and substrate specificity. The enzyme has a relative molecular mass of 105000 and consists of three identical subunits. The purified enzyme showed higher activity and affinity toward cyclic imides, such as succinimide (Km = 0.94 mM: Vmax = 910 µmol -min-1 mg-1), glutarimide (Km, = 4.5 mM: Vmax = 1000 µmol . min-1 mg-1) and maleimide (Km = 0.34 mM: Vmax = 5800 µmol min-1 . mg-1), than toward cyclic urcides, which arc the substrates of dihydropyrimidinases, such as dihydrouracil and hydantoin. Sulfur-containing cyclic imides, such as 2.4-thiazolidinedione and rhodanine. were also hydrolyzed. The enzyme catalyzed the reverse reaction, cyclization, but with much lower activity and affinity. The enzyme was non-competitively inhibited by succinate, which was found to be a key compound in cyclic-imide transformation in relation with the tricarboxylic acid cycle in this bacterium, suggesting that the role of imidase is to catalyze the initial step of cyclic-imide degradation. [ABSTRACT FROM AUTHOR]

A Δ5 and Δ12 desaturase-defective mutant of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, produced 8,11,14,17- cis-eicosatetraenoic acid (20:4ω3) intracellularly when grown with linseed oil. Dihomo-γ-linolenic acid was the only C20 polyunsaturated fatty acid (4.9 wt% of total mycelial fatty acids) other than 20:4ω3. AA and 5,8,11,14,17- cis-eicosapentaenoic acid were not detected. The mycelial lipids consisted of 82.2% (by mol) triacylglycerol (TG), 7.1% diacylglycerol, 8.9% phospholipids (PL), and 1.9% free fatty acids. The percentage of 20:4ω3 was higher in PL (30.1%) than in TG (11.6%), and highest in phosphatidylcholine (38.9%). Under the optimal conditions with a 5-L jar fermenter, 20:4ω3 production amounted to 97.4 mg/g dry mycelia with a mycelial yield of 23 g/L on the twelfth day (corresponding to 2.24 g/L medium and 37.1% of total mycelial fatty acids). [ABSTRACT FROM AUTHOR]