Solid-state cultivation of multiple industrial strains of koji mold on different Thai unpolished rice cultivars: biotransformation of phenolic compounds and their effects on antioxidant activity.

Colored rice is abundant in polyphenols, and koji molds have potential for biotransformation. This study aimed to produce Thai-colored rice koji to study its polyphenolic biotransformation. Four industrial koji mold strains: Aspergillus oryzae 6001, A. oryzae 6020, A. sojae 7009, and A. luchuensis 8035, were cultivated on unpolished Thai-colored rice (Riceberry and Sangyod), unpolished Thai white rice (RD43), and polished Japanese white rice (Koshihikari). We discovered that koji molds grew on all the rice varieties. Methanol extracts of all rice kojis exhibited an approximately 2-fold or greater increase in total phenolic content and DPPH antioxidant activity compared to those of steamed rice. Moreover, quercetin, quercetin-3-O-glucoside, isorhamnetin-3-O-glucoside, ferulic acid, caffeic acid, protocatechuic acid, vanillic acid, (+)-catechin, and (-)-epicatechin content increased in Riceberry and Sangyod koji samples. Consequently, Aspergillus solid-state cultivation on unpolished Thai-colored rice exhibited higher functionalization than the cultivation of unpolished Thai white rice and polished Japanese white rice.

Comparison of taste characteristics between koji mold-ripened cheese and Camembert cheese using an electronic tongue system.

Koji mold, classified in the genus Aspergillus, is used to produce traditional Japanese fermented foods such as miso, soy sauce, and sake. In recent years, the application of koji mold to cheese ripening has attracted attention, and cheese surface-ripened with koji mold (koji cheese) has been studied. In this study, to evaluate the taste characteristics of koji cheese, an electronic tongue system was employed to measure the taste values of cheese samples ripened using 5 strains of koji mold in comparison with commercial Camembert cheese. All koji cheese samples exhibited lower sourness and greater bitterness, astringency, saltiness, and umami richness than the Camembert cheese samples. The intensity of each taste characteristic differed depending on the koji mold strain. These results indicate that koji cheese has a different taste value than conventional mold-ripened cheese. Furthermore, the results also indicate that various taste characteristics can be achieved by selecting different koji molds.

Effect of sake lees on cheese components in cheese ripened by Aspergillus oryzae and lactic acid bacteria.

More than 2,000 varieties of cheese currently exist in the world, and cheese manufacture continues to flourish. To develop the cheese ripening process, additional ingredients are used during cheese production. In this study, the effect of sake lees as an additional ingredient on the fermentation of cheese using Aspergillus oryzae (koji mold), known as koji cheese, was investigated. Aspergillus oryzae is used in the fermentation of Japanese traditional foods, such as sake and soy sauce, given its strong enzymatic activities, as well as in cheese production (i.e., koji cheese). Sake lees, a by-product of the fermentation of rice with A. oryzae and yeasts in the sake brewing process, contains various metabolites, such as amino acids. Here, supplementation with sake lees enhanced the activities of lactic acid bacteria and affected the color of the cheese. Metabolome analysis revealed that sake lees altered the balance of carbohydrates and fatty acids in the cheese. Remarkably, supplementation with sake lees enhanced the production of umami-enhancing γ-glutamyl (kokumi-active) peptides. This study suggests that a new type of cheese can be produced using A. oryzae and sake lees, and information on the synergistic effects of A. oryzae and sake lees will aid the development of cheese production.

[Current Status of Prostate Cancer and Its Relation to Home Medical Care].

Because ofboth the indolent and aggressive nature ofprostate cancers, it is not easy to select the best treatment for patients receiving home medical care who already have many diseases. Since the growth ofprostate cancer is generally slow and all treatments adversely affect the quality of life to some degree, conservative treatment may well be the best option for these patients with prostate cancer. However, it is also true that we often encounter home medical care patients with a locally advanced cancer who had symptoms such as difficulty to urine, macrohematuria and/or lumbago. Such patients need to be diagnosed soon and treated with mainly hormonal therapy after consultation with the specialist. Thus, after a careful evaluation ofthe nature ofthe cancer and comorbidity, we need to provide the best option ofdiagnosis and treatment for home medical care patients to maintain their quality of life.

Effectiveness of a Self-monitoring Device for Urinary Sodium-to-Potassium Ratio on Dietary Improvement in Free-Living Adults: a Randomized Controlled Trial.

BACKGROUND: Reducing the urinary sodium-to-potassium ratio is important for reducing both blood pressure and risk of cardiovascular disease. Among free-living Japanese individuals, we carried out a randomized trial to clarify the effect of lifestyle modification for lowering urinary sodium-to-potassium ratio using a self-monitoring device. METHODS: This was an open, prospective, parallel randomized, controlled trial. Ninety-two individuals were recruited from Japanese volunteers. Participants were randomly allocated into intervention and control groups. A month-long dietary intervention on self-monitoring urinary sodium-to-potassium ratio was carried out using monitors (HEU-001F, OMRON Healthcare Co., Ltd., Kyoto, Japan). All participants had brief dietary education and received a leaflet as usual care. Monitors were handed out to the intervention group, but not to the control group. The intervention group was asked to measure at least one spot urine sodium-to-potassium ratio daily, and advised to lower their sodium-to-potassium ratio toward the target of less than 1. Outcomes included changes in 24-hour urinary sodium-to-potassium ratio, sodium excretion, potassium excretion, blood pressure, and body weight in both groups. RESULTS: Mean measurement frequency of monitoring was 2.8 times/day during the intervention. Changes in urinary sodium-to-potassium ratio were -0.55 in the intervention group and -0.06 in the control group (P = 0.088); respective sodium excretion changes were -18.5 mmol/24 hours and -8.7 mmol/24 hours (P = 0.528); and corresponding potassium excretion was 2.6 mmol/24 hours and -1.5 mmol/24 hours (P = 0.300). No significant reductions were observed in either blood pressure or body weight after the intervention. CONCLUSIONS: Providing the device to self-monitor a sodium-to-potassium ratio did not achieve the targeted reduction of the ratio in "pure self-management" settings, indicating further needs to study an effective method to enhance the synergetic effect of dietary programs and self-monitoring practice to achieve the reduction. However, we cannot deny the possibility of reducing sodium-to-potassium ratio using a self-monitoring device.

[Solitary Fibrous Tumor of the Kidney].

A 59-year-old woman was referred to our hospital with the finding of an incidentaloma in her right kidney. Computed tomographyand magnetic resonance imaging showed a solid tumor, approximately8 cm in diameter, at the lower pole of the right kidney. Partial nephrectomy was performed without a renal vessel clamp. Histopathologically, fibroblast-like spindle cells were proliferating with rich collagenous fibers, and tumor cells were infiltrated into the renal parenchyma. The tumor cells were stained positive with CD34, CD99, and bcl-2. The tumor was diagnosed as solitaryfibrous tumor originating from the kidney based on these histopathological findings. The patient was followed 3 years after surgery with neither recurrence nor metastasis.

Analysis of nitrogen source assimilation in industrial strains of Aspergillus oryzae.

Nitrogen source assimilation is important for the biological functions of fungi, and its pathway has been deeply studied. Aspergillus oryzae mutants defective in nitrogen source assimilation are known to grow poorly on Czapek-Dox (CD) medium. In this study, we found an industrial strain of A. oryzae that grew very poorly on a CD medium containing sodium nitrate as a nitrogen source. We used media with various nitrogen components to examine the steps affecting the nitrogen source assimilation pathway of this strain. The strain grew well on the CD medium supplied with nitrite salt or ammonium salt, suggesting that the strain was defective in nitrate assimilation step. To ascertain the gene causing the defect of nitrate assimilation, a gene expression vector harboring either niaD or crnA of A. oryzae RIB40 was introduced into the industrial strain. The industrial strain containing the crnA vector recovered its growth. This is the first report that a mutation of crnA causes poor growth on CD medium in an industrial strain of A. oryzae, and crnA can be used as a transformation marker for crnA deficient strains.

Multifunctional-separation-mode ion chromatography method for determining major metabolites during multiple parallel fermentation of rice wine.

Facile and effective analysis methods are desirable for elucidating the behaviours of metabolites during fermentation reactions. Herein, a multifunctional-separation-mode ion chromatography (MFS-IC) method was developed for the simultaneous monitoring of major metabolites during multiple parallel fermentation, including those related to central carbon metabolism (saccharification, glycolysis, alcoholic fermentation, and the tricarboxylic acid (TCA) cycle). The use of two types of sulfo-modified size-exclusion columns and phthalic acid as the eluent allowed the separation of oligosaccharides (disaccharides, trisaccharides, and tetrasaccharides), glucose, pyruvate, and major organic acids during the TCA cycle (cis-aconitate, citrate, iso-citrate, malate, fumarate, and succinate but not α-ketoglutarate) from other non-target analytes. The MFS-IC method was successfully applied to monitoring the major metabolites in the rice wine brewing process. This approach can contribute to an improved understanding of metabolite behaviour during fermentation without requiring the use of expensive advanced instrumentation methods such as liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.

Preparation of egg-koji for developing a novel food.

While chicken eggs contain many nutrients necessary for humans and there are various cooking methods, the nutritional components are used as they are, and there are no traditional foods that utilize microorganisms. Koji-mold, containing Aspergillus oryzae, A. sojae, and A. luchuensis, which has been used in various fermented foods since ancient times, grows on raw grain materials such as rice and barley to become koji. This can give flavors not found in the raw materials that can decompose and convert the nutritional components of the raw materials. Here, we succeeded for the first time in developing egg-koji that uses only eggs and koji-mold by selecting and combining cooked egg powder (CEP) and A. oryzae AO101 as the most suitable combination. To suppress the explosive growth of harmful bacteria, we improved the sterilization method, watering method, and amount of water. In addition, it was found that egg-koji has a characteristic enzyme activity balance, in which amylase is extremely low and protease at pH 6 was high compared to grain koji, such as rice and barley. Egg-koji might produce enzymes suitable for taking in nutrients when growing into CEP and would be expected to give a flavor that could not be achieved by cooking or additives.

Volatile and soluble metabolite profiles in surface-ripened cheeses with Aspergillus oryzae and Aspergillus sojae.

To determine the impact of traditional koji molds on chemical characteristics of soft-type natural cheese, novel surface mold-ripened cheeses with Aspergillus oryzae and Aspergillus sojae were studied by non-targeted metabolite profiling. Comprehensive water-soluble and volatile metabolite profiles of koji cheese were evaluated among five Aspergillus strains and other mold-ripened cheeses. Time-course changes in the metabolite profiles and degrading enzyme activities were also compared with those of an industrial Penicillium candidum starter culture. Koji cheeses differed from Camembert, Brie, and blue cheeses in higher lactic acid, amino acid, and acetoin levels and lower methyl ketone and volatile fatty acid levels. Time-course analysis revealed the associations of rapid accumulations of glutamic, aspartic, and 3-methylbutanoic acids and 3-methylbutanal with higher proteolytic activity, and methyl ketone and fatty acid derivative suppressions with lower lipolytic activity. Ethyl butanoate, diacetyl, and malic acid also characterized koji cheeses as strain-dependent metabolites. This study highlighted the key compositional difference derived from cheese ripening with Aspergillus strains. The findings could help quality improvements of koji cheese product.

Koji Starter and Koji World in Japan.

Koji is made by culturing koji mold on grains. Koji has wide-ranging applications, for example, in alcoholic beverages and seasonings. The word 'mold' generally has a bad image, but in Japan, koji mold is valued for its usefulness, and over the years, efforts have been made to make safe, stable, and delicious food products from it. Koji mold spores, essential when making koji, are called koji starter in the industry. From the many available strains, those suitable for the production of each fermented food are chosen based on indicators such as growth rate and enzyme production capacity. In manufacturing using microorganisms, purity and yield are prioritized. However, the production of fermented foods using koji is more complex, with focus not only on the degree of decomposition of raw materials but also on factors influencing overall product design, including palatability, color, smell, and texture. Production can be facilitated by the variety of koji brought about by the diversity of koji mold combined with the solid culture method which increases the amount of enzyme production. In this report, we introduce the history of koji starter in Japan, the characteristics of koji mold in practice, and various fermented foods made from it. In addition, the factors affecting the quality of koji in solid culture are described.

Single assistant versus dual assistant robotic surgery for robot-assisted laparoscopic hysterectomy using da Vinci Xi or X.

BACKGROUND: To verify the usefulness of single assistant robotic surgery (SA-RS) using da Vinci Xi or X performed for total hysterectomy. METHODS: The SA-RS group (S-group) includes surgeries in which the surgeon performed all surgical operations other than that of the uterine manipulator, while the dual assistant robotic surgery group (D-group) includes surgeries performed by the surgeon with two assistants. In S-group, the forceps and camera were replaced during surgery in cases of a large uterus with limited range of motion of the instrument. A comparative study of patient background, intraoperative and postoperative results and surgery-related cost was performed between the two groups. RESULTS: No significant differences were observed between the two groups. On the contrary, S-group showed a significant reduction in preparation time and wound closure time, and a savings of $768. CONCLUSION: SA-RS for total hysterectomy was possible, which is excellent in terms of cost, esthetics, and manpower.

Rapid noninvasive monitoring of freshness variation in frozen shrimp using multidimensional fluorescence imaging coupled with chemometrics.

Shrimp is one of the most delicious and popular food commodities worldwide due to its exceptional taste and characteristics. Freshness is considered as a key factor for shrimp consumers because freshness has a significant relationship with taste and shelf-life of shrimp. However, post-mortem metabolism of shrimp differs from that of fish as they are highly susceptible to post-harvest quality loss, and it is hard to distinguish the freshness variation of shrimp at frozen state instantly. Thus, instant monitoring of frozen shrimp freshness is challenging for the seafood and aquaculture industries and a reliable, expeditious, and noninvasive technique to estimate shrimp quality is in high demand. Accordingly, this study aimed to visualize changes in post-mortem freshness of frozen shrimp using multidimensional fluorescence imaging. Live coonstripe shrimp (Pandalus hypsinotus) were harvested and instantly killed by beheading, cooled on ice for 0, 6, 24, 48, 72 and 96 h (n = 8), followed by processing into frozen peeled deveined shrimp product and stored at -60 °C. 50% of frozen shrimp were analyzed for excitation-emission matrix (EEM), ATP-related compounds, and pH using a fiber optic supported fluorescence spectrophotometer (F-7100), high performance liquid chromatography (HPLC) and pH meter, respectively at each time point (n = 4). Then, fluorescence images were obtained from the remaining 50% of frozen shrimp (n = 4) by computer vision method equipped with a charge-coupled device (CCD) camera, MAX-303 xenon light source for an excitation light (Ex. 330 nm), and an automatic filter changer for emission band-pass filters (Em. 380-610 nm at 10 nm intervals). Chemical analysis of frozen shrimp revealed that K-value and pH of shrimp increased from 1.61 to 66.56% and 6.49-7.31, respectively, during storage on ice. Repeated partial least squares regression (PLSR) models of EEM for K-value prediction suggested an efficient excitation wavelength (330 nm) and its corresponding emission wavelengths (380-610 nm) to produce fluorescence images. Spatial-temporal changes of K-value and pH were visualized successfully in frozen shrimp by fluorescence imaging. K-value visualization was then validated effectively using another group of frozen shrimp (0-72 h ice stored) with different killing method (super chilling) and the prediction accuracy was R2 = 0.80. This novel approach using a CCD camera coupled with EEM provides a state-of-the-art authentication method for practical assessment of frozen seafood freshness.

Prostate carcinoma with neuroendocrine differentiation successfully treated by early detection with imaging examination.

INTRODUCTION: Clinicopathological features of neuroendocrine differentiation of prostate carcinoma have not been totally clarified yet. It is known to be associated with poor prognosis. CASE PRESENTATION: A patient with growing prostate-specific antigen was diagnosed with prostate carcinoma and treated by laparoscopic prostatectomy. The pathological examination revealed the neuroendocrine differentiation of the tumor. Early detection of prostate carcinoma with neuroendocrine differentiation can be difficult due to its low expression of prostate-specific antigen. The imaging examination contributed to the early detection. In the follow-up period of 2 years, the patient remains cancer free. CONCLUSION: Recently, the treatment options for prostate carcinoma have been expanded. Precise assessment of immunohistochemical nature of the tumor may be helpful for individualized decision-making.

Gleason Pattern 5 is a Possible Pathologic Predictor for Biochemical Recurrence after Laparoscopic Radical Prostatectomy

Objective: Several prognostic factors for biochemical recurrence after radical prostatectomy have been reported, including initial prostate-specific antigen level, Gleason score, positive surgical margin, and seminal vesicle invasion. Here we investigate whether Gleason pattern 5 is a predictor for biochemical recurrence. Methods: This retrospective study included 168 patients who underwent laparoscopic radical prostatectomy from 2006 to 2015. The relationship between biochemical recurrence after laparoscopic radical prostatectomy and the presence of Gleason pattern 5, even as a tertiary pattern, was investigated. Biochemical recurrence was defined when the prostate-specific antigen level rose to >0.2 ng/ml after having decreased to <0.1 ng/ml following laparoscopic radical prostatectomy. Biochemical recurrence-free survival was estimated by the Kaplan-Meier method. Multivariate analysis was performed using a Cox proportional hazards regression model. Results: The median age was 66 years, median initial prostate-specific antigen level was 6.9 ng/ml, and median follow-up period was 47.3 months. Biochemical recurrence was recognized in 27 patients (16.1%) after laparoscopic radical prostatectomy, and 5-year biochemical recurrence-free survival was 78.6%. Gleason pattern 5 was noted in 5 patients as the primary pattern, in 10 as the secondary pattern, and in 5 as the tertiary pattern. According to multivariate analysis, presence of Gleason pattern 5 (HR = 4.75, p=0.001) and positive surgical margin (HR = 4.66, p=0.001) were independent predictive factors for biochemical recurrence-free survival. Conclusion: Gleason pattern 5 appears to be an important predictive factor for biochemical recurrence after laparoscopic radical prostatectomy.

[Incidental inverted papilloma during the clinical follow-up of superficial transitional cell carcinoma in the urinary bladder].

A 76-year-old woman presented with gross hematuria. She had received transurethral resections (TURBT) twice for superficial bladder tumors near the right orifice. All pathologic findings demonstrated low grade superficial transitional cell carcinoma (TCC). Cystoscopy showed the tumor lesions during the follow-up. Bladder tumors were completely harvested and pathologic examination revealed inverted papilloma. However, recurrent tumors were detected at the same location after 4 months' follow up. Although she came to our hospital for the purpose of TURBT, she complained of severe neck pain and was suddenly dead. Autopsy findings showed that dissecting aortic aneurism was the cause of death and the bladder tumor was low grade superficial TCC. The case reported here had a very rare pathologic finding with a history of superficial tumors located in the same areas in the urinary bladder.

[Metastatic small intestinal tumor associated with transitional cell carcinoma: a report of 2 cases and review of cases in Japan].

Transitional cell carcinoma (TCC) frequently metastasizes to lymph nodes, liver, lungs and bone. However, metastasis to the gastrointestinal tract is rare. We report two cases of bladder tumor which metastasized to the ileum. According to the literature, these are the 7th and 8th cases in Japan. Case 1: A 87-year-old man had a history of bladder tumor (TCC, grade 3, pT2bN0M0) and has transurethral resection of bladder tumor (TUR-BT) three times. Two months after the last TUR-BT, he was admitted with ileus. As computed tomography (CT) showed abdominal free air, our diagnosis was perforation of gastrointestinal tract. The patient received an operation which resected partial ileum. We found the elastic hard tumor in the ileum on the perforated lesion, which showed metastatic TCC in the ileum pathologicaly. Case 2 : A 53-year-old man visited our hospital with gross hematuria. Cystoscopy showed a non-papillary broad based tumor in the right wall of the bladder. CT showed a bladder tumor invaded into the prostate (pT4aN1M0), we performed total cyctectomy and ileal conduit after neo-adjuvant chemotherapy. During the operation, we found the tumor (2 cm in diameter) in the small intestine which was metastasized of bladder tumor.

Phase Transition during Heating of Nanostructured Ultrahigh Molecular Weight Polyethylene Membranes.

Ultrahigh molecular weight polyethylene (UHMW-PE) membranes were prepared using biaxial melt-drawing and subsequent melt-shrinking. Electron microscopy observations indicate that the former membrane has more extended-chain crystals (ECCs), whereas the latter is mainly composed of folded-chain crystals (FCCs). Corresponding double-melting endotherms are recorded on differential scanning calorimetry (DSC) measurements. Detailed assignments of such double-melting components are performed using in situ X-ray measurements during heating. Wide- and small-angle X-ray diffraction and scattering (WAXD/SAXS) images were simultaneously recorded at SPring-8. Changes in WAXD images indicate that the orthorhombic reflection peak begins to decrease at 130 °C, followed by the appearance of the hexagonal reflection peak beyond 145 °C for both membranes, but the latter melt-shrunk membrane exhibits weaker hexagonal reflection intensity. Simultaneous SAXS results indicate that FCCs rapidly disappear at 135 °C for the melt-shrunk membrane, resulting in a sharper endotherm. In contrast, residual ECCs restrict the melting of FCCs for the melt-drawn membrane, resulting in a broader endotherm of FCC melting spread to a slightly higher temperature position.

[A case of squamous metaplasia of the ureter].

This paper reports a 73-year-old man who developed ureteral squamous metaplasia. Preoperative drip infusion pyelography showed a ureteral tumor with a major axis measuring 2.5 cm, which had a smooth surface and a broad base. Computed tomography findings suggested that the lesion might be a submucous mesodermal tumor or inverted papillary transitional cell carcinoma. Therefore, retroperitoneal laparoscopic left nephroureterectomy was performed.

[Experience with the memotherm stent in high-risk urinary retention patients].

PURPOSE: We report on our experience with the Memotherm stent implanted in high-risk patients who had an indwelling catheter. PATIENTS AND METHODS: Seventeen patients (mean age 80.6 years, range 55 to 93) who had an indwelling catheter were treated by placement of the Memotherm stent. The mean duration time of indwelling catheter were 12.7 months (range 1 to 60). The methods of implantation was used the special delivery system and treatment can be done with the patient under local anesthesia with non-steroidal anti-inflammatory drug. To assess the efficacy of the treatment we measured urinary peak flow rate (ml/sec), residual urine volume (ml) and residual urine rate (residual urine volume/total urine volume: %). Subjective symptoms were evaluated using the International Prostate Symptom Score (I-PSS). The mean follow-up time was 3.3 months (range 1 week to 8 months). RESULTS: There were no severe medical problems associated with stent placement. Sixteen patients were able to void immediately after insertion of the stent. The remaining 1 required an indwelling catheter to 2 months, however, eventually able to void. Only 7 of the 17 patients can be evaluation for urinary flow and residual urine, nevertheless the mean urinary peak flow rate showed 9.7 ml/sec, the mean residual urine volume was 77.4 ml, and the mean residual urine rate was 27.5%. Also, average I-PSS showed 13.4 in these patients. CONCLUSIONS: As a results of this study, the Memotherm stent is good conservative therapy for high-risk urinary retention patients.