Three-dimensional dynamics of fluctuations appearing during pellet ablation process around a pellet in a fusion plasma experiment.

Understanding pellet ablation physics is crucial to realizing efficient fueling into a high temperature plasma for the steady state operation of ITER and future fusion reactors. Here we report the first observation of the formation of fluctuation structures in the pellet plasmoid during the pellet ablation process by a fast camera in a medium-sized fusion device, Heliotron J. The fluctuation has a normalized fluctuation level of ~ 15% and propagates around the moving pellet across the magnetic field. By comparing the fluctuation structures with the shape of magnetic field lines calculated with the field line tracing code, we successfully reconstruct the spatio-temporal structure of the fluctuations during the pellet ablation process. The fluctuations are located at the locations displaced toroidally from the pellet and propagate in the cross-field direction around the pellet axis along the field line, indicating a three-dimensional behavior and structure of fluctuations. The fluctuation would be driven by a strong inhomogeneity formed around the pellet and invoke the relaxation of the gradient through a cross-field transport induced by the fluctuations, which could affect the pellet ablation and pellet fueling processes. Such fluctuations can be ubiquitously present at the inhomogeneity formed around a pellet in the pellet ablation process in fusion devices.

Lactic Acid Bacteria in Total Mixed Ration Silage Containing Soybean Curd Residue: Their Isolation, Identification and Ability to Inhibit Aerobic Deterioration.

We investigated the effects of the predominant lactic acid bacteria (LAB) on the fermentation characteristics and aerobic stability of total mixed ration (TMR) silage containing soybean curd residue (SC-TMR silage). The SC-TMR materials were ensiled in laboratory silos for 14 or 56 days. LAB predominant in SC-TMR silage were identified (Exp. 1). Lactobacillus fermentum (L. fermentum) and Streptococcus bovis (S. bovis) were found in the untreated materials, Leuconostoc pseudomesenteroides (L. pseudomesenteroides) in 14-day silage and Lactobacillus plantarum (L. plantarum) in all silages. Pediococcus acidilactici (P. acidilactici), Lactobacillus paracasei (L. paracasei), and Lactobacillus brevis (L. brevis) formed more than 90% of the isolates in 56-day silage. Italian ryegrass and whole crop maize were inoculated with P. acidilactici and L. brevis isolates and the fermentation and aerobic stability determined (Exp. 2). Inoculation with P. acidilactici and L. brevis alone or combined improved the fermentation products in ryegrass silage and markedly enhanced its aerobic stability. In maize silage, P. acidilactici and L. brevis inoculation caused no changes and suppressed deterioration when combined with increases in acetic acid content. The results indicate that P. acidilactici and L. brevis may produce a synergistic effect to inhibit SC-TMR silage deterioration. Further studies are needed to identify the inhibitory substances, which may be useful for developing potential antifungal agents.

Identification and isolation of Lactobacillus fructivorans from wilted alfalfa silage with and without molasses.

AIM: To gain insights into the bacterial species associated with anaerobic storage and aerobic stability of alfalfa silage. METHODS AND RESULTS: Wilted alfalfa silage (498 g dry matter kg(-1) ) was prepared with and without the addition of molasses. Aerobic spoilage tests were conducted at 5, 10 and 60 days after ensiling. The composition of fermentation products and the bacterial communities of silage were determined at 1, 3, 5 and 7 days after silo opening. Silage without molasses had small amounts of lactic and acetic acids detectable at silo opening but resisted deterioration due to aerobic spoilage for at least 5 days after opening. Resistance to aerobic deterioration in silage increased with the addition of molasses. The predominant bacterial species in molasses-added silage was Lactobacillus fructivorans, which was detected by denaturing gradient gel electrophoresis analysis. Different bacterial growth media were used for Lact. fructivorans isolation from alfalfa silage with added molasses: isolation was successful using liver infusion sake medium, but was unsuccessful when de Man, Rogosa and Sharpe medium was used. CONCLUSION: A nonconventional lactic acid bacterium (LAB) species may be involved in the high aerobic stability of alfalfa silage. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings demonstrate that culture-independent microbiota analysis may be useful in the isolation and identification of nonconventional LAB species involved in fermentation and the aerobic stability of silage.

Dynamical programming based turbulence velocimetry for fast visible imaging of tokamak plasma.

An orthogonal dynamic programming (ODP) based particle image velocimetry (PIV) technique is developed to measure the time resolved flow field of the fluctuating structures at the plasma edge and scrape off layer (SOL) of tokamaks. This non-intrusive technique can provide two dimensional velocity fields at high spatial and temporal resolution from a fast framing image sequence and hence can provide better insights into plasma flow as compared to conventional probe measurements. Applicability of the technique is tested with simulated image pairs. Finally, it is applied to tangential fast visible images of QUEST plasma to estimate the SOL flow in inboard poloidal null-natural divertor configuration. This technique is also applied to investigate the intricate features of the core of the run-away dominated phase following the injection of a large amount of neutrals in the target Ohmic plasma. Development of the ODP-PIV code and its applicability on actual plasma images is reported.

Identification of lactic acid bacteria in the rumen and feces of dairy cows fed total mixed ration silage to assess the survival of silage bacteria in the gut.

The survival of silage lactic acid bacteria (LAB) in the gut of dairy cows was evaluated by examining the LAB communities of silage and gut contents. Samples were collected at 2 different research institutes (Mie and Okayama) that offered total mixed ration (TMR) silage throughout the year. Silage and feces were sampled in August, October, and November at the Mie institute, whereas silage, rumen fluid, and feces were sampled in June and August at the Okayama institute. Denaturing gradient gel electrophoresis using Lactobacillus-specific primers was performed to detect LAB species in the samples. The selected bands were purified for species identification and the band patterns were used for principal component analysis. Lactic acid was the predominant fermentation product in all the TMR silages analyzed, and the lactic acid level tended to be constant regardless of the sampling time and region. A total of 14 LAB species were detected in the TMR silage samples, of which 5 (Lactobacillus acetotolerans, Lactobacillus pontis, Lactobacillus casei, Lactobacillus suebicus, and Lactobacillus plantarum) were detected in the dairy cow feces. Most of the denaturing gradient gel electrophoresis bands for the feces samples were also detected in the rumen fluid, suggesting that any elimination of silage LAB occurred in the rumen and not in the postruminal gut segments. The principal component analysis indicated that the LAB communities in the silage, rumen fluid, and feces were separately grouped; hence, the survival of silage LAB in the cow rumen and lower gut was deemed difficult. It was concluded that, although the gut LAB community is robust and not easily affected by the silage conditions, several LAB species can inhabit both silage and feces, which suggests the potential of using silage as a vehicle for conveying probiotics.

Effects of storage temperature and ensiling period on fermentation products, aerobic stability and microbial communities of total mixed ration silage.

AIMS: To examine how storage temperatures influence ensiling fermentation, aerobic stability and microbial communities of total mixed ration (TMR) silage. METHODS AND RESULTS: Laboratory-scale silos were stored at 5, 15, 25 and 35°C for 10, 30 and 90 days. If silage was stored at 5°C, fermentation was weak until day 30, but acceptable lactic acid production was observed on day 90. The ethanol content was higher than the acetic acid content when stored at 15 and 25°C, whereas the ethanol content was lower when stored at 35 than at 25°C. Aerobic deterioration did not occur when silage was exposed to air at the same temperature at which it was stored. Although 10-day silages stored at 5 and 15°C deteriorated when the aerobic stability test was conducted at 25°C, heating was not observed in silages stored at 25 or 35°C or in any 90-day silages regardless of storage temperature. Denaturing gradient gel electrophoresis demonstrated that bands indicative of Lactobacillus plantarum and Lactobacillus delbrueckii were less prominent, while bands indicative of Lactobacillus panis became more distinct in silages stored at high temperatures. Bands of Kluyveromyces marxianus were seen exclusively in silages that were spoiled at 25°C. CONCLUSION: High ambient temperature enhances acetic acid production in TMR silage. Lactobacillus panis may be associated with changes in the fermentation products due to differences in storage temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: The role of Lacto. panis in ensiling fermentation and aerobic stability is worth examining.

NK cell-based autologous immune enhancement therapy (AIET) for cancer.

Natural killer (NK) cells have been known to enhance the host immune responses against cancer. NK cell number and cytotoxicity in patients with cancer is often low. Therefore, we developed a large-scale ex vivo NK cell expansion method without feeder layers and introduced NK cell-based autologous immune enhancement therapy (AIET). In this paper, we discuss the epidemiological data that show the relationship between NK activity and cancer incidence, monitoring of NK cell number and activity, anti-cancer activities of NK cells in vitro and in vivo and the effects of the combination of expanded NK cells with monoclonal antibody drugs on cancers through antibody-dependent cellular cytotoxicity. Finally, we also present the clinical cases of NK cell-based AIET and the effect of AIET on advanced stage of pancreatic cancer and on various advanced cancers refractory to conventional therapies. NK cell-based AIET might be a useful strategy in the multidisciplinary approach to cancer.

Fast visible imaging and edge turbulence analysis in QUEST.

A fast visible imaging system is installed on the spherical tokamak QUEST to study edge turbulence. The camera uses a complementary metal-oxide semiconductor detector with a maximum resolution of 1024 × 1024 at 7000 frames∕s (fps) and can achieve 775 kfps at a resolution of 128 × 24. In this paper, we present the salient features of the system and its application to study edge turbulence in 8.2 GHz ECRH driven slab plasma, without plasma current. Vertical magnetic field (B(z)) topology is varied with three sets of poloidal field (PF) coils and the variation in the edge turbulence is investigated as a function of the B(z) strength and curvature. Fluctuation amplitude was highest for the shallow PF well. Cross-correlation coefficient shows distinct coherent mode along z direction at the steep density gradient region and it grows with the PF mirror ratio.

Survival of silage lactic acid bacteria in the goat gastrointestinal tract as determined by denaturing gradient gel electrophoresis.

AIMS: To determine the survival rate of silage lactic acid bacteria (LAB) in the ruminant gastrointestinal tract. METHODS AND RESULTS: Wilted Italian ryegrass (Lolium multiflorum Lam.) silage (containing 1·9×10(6) CFU LAB g(-1)) was fed ad libitum to three goats equipped with rumen cannulae. Silage was given alone or with concentrates at a 1:1 ratio on a dry matter basis. Rumen fluid was then obtained 2, 4 and 8h after the morning feeding. Denaturing gradient gel electrophoresis was performed to compare LAB communities in silage, rumen fluid and faeces. The LAB detected in the wilted silage included Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus murinus and Lactobacillus sakei. Bands indicative of Lact. murinus were detected in either the rumen fluid or faeces, whereas the bands indicative of Lact. plantarum, Lact. brevis and Lact. sakei were not. Although the rumen fluid LAB counts and volatile fatty acid concentrations were higher in goats fed silage plus concentrates compared with those fed silage alone, the LAB communities themselves remained unaffected. Sampling times and goat-to-goat variations did not affect the LAB communities found in the rumen fluid. CONCLUSION: LAB communities found in the gut are not remarkably affected by the consumption of silage LAB, even when the silage is accompanied by concentrates that facilitate gut fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: Although silage can improve probiotic function, it may be difficult for silage LAB to survive the digestive process in the ruminant gastrointestinal tract.

Effects of wilting and molasses addition on fermentation and bacterial community in guinea grass silage.

AIMS: Acetic acid is considered an important preservative in tropical grass ensiling. The objective of the current experiments was to follow the ensiling fermentation of low dry matter (DM) tropical grass as a model to study changes in bacterial communities during acetic acid fermentation. METHODS AND RESULTS: Direct-cut and wilted guinea grass silage was prepared with and without molasses. A high acetic acid level was observed during the fermentation of direct-cut silage, and long storage increased the butyric acid and ethanol content if molasses was not added. The lactic acid production in wilted silage was greater than the acetic acid production, but prolonged ensiling decreased the lactic to acetic acid ratio regardless of molasses addition. Adding molasses enhanced the lactic acid content in both direct-cut and wilted silage. The bacterial community, identified by denaturing gradient gel electrophoresis, was affected by wilting and molasses addition. Bands for Pantoea sp. and Morganella sp. became faint when acetic acid fermentation was suppressed, and those for Pediococcus pentosaceus and Lactococcus garvieae were detected when lactic acid fermentation was enhanced by wilting and molasses addition. Lactobacillus plantarum and Lactococcus lactis were found throughout the ensiling process in all silage types. CONCLUSION: Distinct changes occurred in the bacterial community in guinea grass silage because of wilting and molasses addition. These changes could explain how lactic acid fermentation was enhanced but could not help determine which bacteria were associated with enhanced acetic acid fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: The study reveals the effects of wilting and molasses during ensiling of low DM tropical grasses and the associated bacteria.

Monitoring the bacterial community of maize silage stored in a bunker silo inoculated with Enterococcus faecium, Lactobacillus plantarum and Lactobacillus buchneri.

AIMS: To monitor variations in the bacterial community and fermentation products of maize silage within and between bunker silos. METHODS AND RESULTS: Silage samples were collected in 2008 and 2009 from three dairy farms, wherein the farmers arranged for a contractor to produce maize silage using bunker silos. Silage was prepared using a lactic acid bacteria (LAB) inoculant consisting of Enterococcus faecium, Lactobacillus plantarum and Lactobacillus buchneri. Eight samples were collected from each bunker silo; 4 'outer' and 4 'inner' samples were collected from near the top and the bottom of the silo. The dry matter, lactic acid, acetic acid, ethanol, 1-propanol and 1,2-propanediol contents differed between bunker silos in both sampling years. Higher acetic acid, 1-propanol and 1,2-propanediol contents were found in the bottom than the top layers in the 2008 samples, and higher lactic acid content was found in the top than the bottom layers in the 2009 samples. The bacterial community varied more between bunker silos than within a bunker silo in the 2008 samples, whereas differences between the top and the bottom layers were seen across bunker silos in the 2009 samples. The inoculated LAB were uniformly distributed, while several nonconventional silage bacteria were also detected. Lactobacillus acetotolerans, Lactobacillus panis and Acetobacter pasteurianus were detected in both years. Stenotrophomonas maltophilia was detected in the 2008 samples, and Lactobacillus reuteri, Acinetobacter sp. and Rahnella sp. were detected in the 2009 samples. CONCLUSIONS: Although differences were seen within and between bunker silos, the bacterial community may indicate a different relationship between bunker silos and sampling locations within a bunker silo from that indicated by the fermentation products. SIGNIFICANCE AND IMPACT OF THE STUDY: Analysis of bacterial community can help understand how diverse non-LAB and LAB species are involved in the ensiling process of bunker-made maize silage.

Bacterial and fungal communities of wilted Italian ryegrass silage inoculated with and without Lactobacillus rhamnosus or Lactobacillus buchneri.

AIMS: To understand the effects of lactic acid bacteria (LAB) inoculation on fermentation products, aerobic stability and microbial communities of silage. METHODS AND RESULTS: Wilted Italian ryegrass was stored in laboratory silos with and without inoculation of Lactobacillus rhamnosus and Lactobacillus buchneri. The silos were opened after 14, 56 and 120 days and then subjected to aerobic deterioration for 7 days. Intensive alcoholic fermentation was found in untreated silage; the sum of ethanol and 2,3-butanediol content at day 14 was about 7 times higher than that of lactic and volatile fatty acids. Alcoholic fermentation was suppressed by L. rhamnosus and L. buchneri inoculation and lactic acid and acetic acid became the dominant fermentation products, respectively. Silages were deteriorated in untreated and L. rhamnosus-inoculated silages, whereas no spoilage was found in L. buchneri-inoculated silage. Enterobacteria such as Erwinia persicina, Pantoea agglomerans and Rahnella aquatilis were detected in untreated silage, whereas some of these bacteria disappeared or became faint with L. rhamnosus treatment. When silage was deteriorated, Lactobacillus brevis and Bacillus pumilus were observed in untreated and L. rhamnosus-inoculated communities, respectively. The inoculated LAB species was detectable in addition to untreated bacterial communities. Saccharomyces cerevisiae and Pichia anomala were the main fungi in untreated and L. rhamnosus-inoculated silages; however, P. anomala was not visibly seen in L. buchneri-inoculated silage either at silo opening or after exposure to air. CONCLUSION: Inoculation with L. rhamnosus can suppress alcoholic fermentation of wilted grass silage with elimination of enterobacteria at the beginning of fermentation. Addition of L. buchneri may improve aerobic stability, with distinct inhibitory effect observed on P. anomala after silo opening. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial and fungal community analyses help us to understand how inoculated LAB can function to improve the fermentation and aerobic stability of silage.

Presence of sourdough lactic acid bacteria in commercial total mixed ration silage as revealed by denaturing gradient gel electrophoresis analysis.

AIMS: To characterize the bacterial communities in commercial total mixed ration (TMR) silage, which is known to have a long bunk life after silo opening. METHODS AND RESULTS: Samples were collected from four factories that produce TMR silage according to their own recipes. Three factories were sampled three times at 1-month intervals during the summer to characterize the differences between factories; one factory was sampled 12 times, three samples each during the summer, autumn, winter and spring, to determine seasonal changes. Bacterial communities were determined by culture-independent denaturing gradient gel electrophoresis. All silages contained lactic acid as the predominant acid, and the contents appeared stable regardless of factories and product seasons. Acetic acid and 1-propanol contents were different between factories and indicated seasonal changes, with increases in warm seasons compared to cool seasons. Both differences and similarities existed among the bacterial communities from each factory and product season. Lactobacillus parabuchneri was found in the products from three of four factories. Various sourdough lactic acid bacteria (LAB) were identified in commercial TMR silage; Lactobacillus panis, Lactobacillus hammesii, Lactobacillus mindensis, Lactobacillus pontis, Lactobacillus frumenti and Lactobacillus farciminis were detected in many products. Moreover, changes owing to product season were distinctive, and Lact. pontis and Lact. frumenti became detectable in summer products. CONCLUSION: Sourdough LAB are involved in the ensiling of commercial TMR silage. Silage bacterial communities vary more by season than by factory. The LAB species Lact. parabuchneri was detected in the TMR silage but may not be essential to the product's long bunk life after silo opening. SIGNIFICANCE AND IMPACT OF THE STUDY: Commercial TMR silage resembles sourdough with respect to bacterial communities and long shelf life. The roles of sourdough LAB in the ensiling process and aerobic stability are worth examining.

Bacterial community associated with ensilage process of wilted guinea grass.

AIMS: To determine the effects of wilting, storage period and bacterial inoculant on the bacterial community and ensiling fermentation of guinea grass silage. METHODS AND RESULTS: Fermentation products, colony counts and denaturing gradient gel electrophoresis (DGGE) profiles were determined. There was more lactic acid than acetic acid in all silages, but the lactic acid to acetic acid ratio decreased with storage time. This shift from lactic to acetic acid was not prevented even with a combination of wilting and bacterial inoculant. The DGGE analyses suggest that facultatively heterofermentative lactic acid bacteria (Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus pentosus) were involved in the shift to acetic acid fermentation. CONCLUSIONS: Lactic acid can dominate the fermentation in tropical grass silage with sufficient wilting prior to ensiling. Prolonged storage may lead to high levels of acetic acid without distinctive changes in the bacterial community. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacterial community looks stable compared to fermentation products over the course of long storage periods in tropical grass silage. Acetic acid fermentation in tropical grass silage can be a result of the changes in bacterial metabolism rather than community structure.

Ensiling of soybean curd residue and wet brewers grains with or without other feeds as a total mixed ration.

Wet brewers grains and soybean curd residue were stored in laboratory-scale silos without (BG and SC silages, respectively) or with other ingredients as total mixed rations (BGT and SCT silages, respectively). Silages were opened after 14 and 56 d, and microbial counts, fermentation products, and aerobic stability were determined. Denaturing gradient gel electrophoresis was carried out to examine bacterial communities, and several bacteria that appeared to be involved in fermentation were identified. Lactic acid content was greater in SCT than in BGT silage, but lower in SC than in BG silage. Ethanol content was greater in BG than in SC regardless of silage type. Aerobic deterioration occurred promptly in ensiling materials (nonensiled by-products and total mixed ration mixtures) and in silages stored alone; however, SCT and BGT silages resisted deterioration and no heating was found for more than 5.5 d regardless of storage period. Silages were stable even with high yeast populations at silo opening, whereas prolonged ensiling decreased yeast counts in the 2 total mixed ration silages. The denaturing gradient gel electrophoresis profiles appeared similar between SCT and BGT silages but not between SC and BG silages. Weissella spp. and Lactobacillus brevis were common in aerobically stable SCT and BGT silages, and Lactobacillus buchneri was detected only in BGT silage. Both L. brevis and L. buchneri were found in silage but not in ensiling materials. Several other lactic acid bacteria were also identified in SCT and BGT silages, but did not appear to be related to fermentation and aerobic stability.

Biogenic amine production in grass, maize and total mixed ration silages inoculated with Lactobacillus casei or Lactobacillus buchneri.

AIMS: To investigate the effects of inoculating Lactobacillus casei or Lacobacillus buchneri on the production of biogenic amines (BA) in silage. METHODS AND RESULTS: Wilted festulolium (Lolium perenne x Festuca pratensis), whole crop maize or a total mixed ration, consisting of wet brewer grains, lucerne hay, cracked maize, sugarbeet pulp, soyabean meal and molasses, was ensiled with or without the inoculation of either L. casei (>10(6) CFU g(-1)) or L. buchneri (>10(6) CFU g(-1)). Silages were opened after 60 days of storage, and the concentrations of histamine, tyramine, putrescine and cadaverine were determined. The inoculation of L. casei decreased all the BA regardless of the silage type. The effects of L. buchneri varied between the three silages; the tyramine and putrescine were increased in maize but were lowered in festulolium. Histamine was reduced in festulolium and the by-products, whereas no change was found in the maize silage. None of the inoculant strains produced the four BA in a synthetic medium, accounting for the actual ensiling except for tyramine and putrescine in maize. CONCLUSIONS: Wide variation would be found in the production of BA owing to the ensiling materials. The inoculation of L. casei can lower the BA concentration, while the effects of L. buchneri may vary considerably. The screening of BA-producing activity may help to reduce the risk of BA contamination in inoculated silage. SIGNIFICANCE AND IMPACT OF THE STUDY: Strains of decarboxylase-negative L. buchneri can enhance the aerobic stability of silage without a concern regarding the large production of putrefactive BA.

Alcoholic fermentation and its prevention by Lactobacillus buchneri in whole crop rice silage.

AIMS: To evaluate the fermentation characteristics and the effects of Lactobacillus buchneri inoculation in ensiling whole crop rice. METHODS AND RESULTS: Laboratory-scale silages were prepared from whole crop rice harvested at yellow-ripe stage. The crop was ensiled for 2 months with and without inoculation of L. buchneri at 10(4), 10(5) and 10(6) CFU g(-1). The effect of prolonged ensiling was also studied by using the same crop; the silos were opened at 1, 3, 6 and 12 months, while the inoculation was made at 10(5) CFU g(-1). Enhanced alcoholic fermentation was found in untreated silage; the sum of ethanol and 2,3-butanediol were seven times higher at 2 months than those of lactic and volatile fatty acids, while the differences were diminished at 12 months owing to the reduction of ethanol in the late ensiling period. Inoculation of L. buchneri inhibited the alcohols; however, ethanol yet prevailed over the fermentation until 6 months, after which acetic acid became the main product in the inoculated silage. Regardless of inoculation and ensiling period, yeasts were not found in whole crop rice silage. CONCLUSIONS: Substantial amounts of ethanol and 2,3-butanediol would be produced in silage prepared from whole crop rice. The alcoholic fermentation can be suppressed when inoculated with L. buchneri. SIGNIFICANCE AND IMPACT OF THE STUDY: Inoculation of L. buchneri could be an option to prevent ethanol fermentation in silage.

Differences in the histological findings, phenotypic marker expressions and genetic alterations between adenocarcinoma of the gastric cardia and distal stomach.

Adenocarcinoma of the gastric cardia (C-Ca) is possibly a specific subtype of gastric carcinoma. The purpose of this study was to clarify the differences in the clinicopathological characteristics between C-Ca and adenocarcinoma of the distal stomach (D-Ca), and also the differences in the expressions of gastric and intestinal phenotypic markers and genetic alterations between the two. The clinicopathological findings in 72 cases with C-Ca were examined and compared with those in 170 cases with D-Ca. The phenotypic marker expressions examined were those of human gastric mucin (HGM), MUC6, MUC2 and CD10. Furthermore, the presence of mutations in the APC, K-ras and p53 genes and the microsatellite instability status of the tumour were also determined. C-Ca was associated with a significantly higher incidence of differentiated-type tumours and lymphatic vessel invasion (LVI) as compared with D-Ca (72.2 vs 48.2%, P=0.0006 and 72.2 vs 55.3%, P=0.0232, respectively). Oesophageal invasion by the tumour beyond the oesophago-gastric junction (OGJ) was found in 56.9% of cases with C-Ca; LVI in the area of oesophageal invasion was demonstrated in 61% of these cases. Also, LVI was found more frequently in cases of C-Ca with oesophageal invasion than in those without oesophageal invasion (82.9 vs 58.1%, P=0.0197). The incidence of undifferentiated-type tumours was significantly higher in cases with advanced-stage C-Ca than in those with early-stage C-Ca (5 vs 36.5%, P=0.0076). A significantly greater frequency of HGM expression in early-stage C-Ca and significantly lower frequency of MUC2 expression in advanced-stage C-Ca was observed as compared with the corresponding values in cases of D-Ca (78.9 vs 52.2%, P=0.0402 and 51.5 vs 84.6%, P=0.0247, respectively). Mutation of the APC gene was found in only one of all cases of C-Ca, and the frequency of mutation of the APC gene was significantly lower in cases of C-Ca than in those of D-Ca (2.4 vs 20.0%, P=0.0108). The observations in this study suggest that C-Ca is a more aggressive tumour than D-Ca. The differences in biological behavior between C-Ca and D-Ca may result from the different histological findings in the wall of the OGJ and the different genetic pathways involved in the carcinogenesis.