Search

This exploratory research examined a four-day interpreter training program in Japan for impacts on participants’ self-reported increases in knowledge and skills. Pre-, during-, and post-training open-ended questionnaires were administered to all 17 participants. Results showed the training program was effective at increasing knowledge of interpretation's definitions, principles, and goals. Participants indicated extended opportunities for understanding the profession as a whole, as well as how to operationalize interpretation and develop strategic outcomes would increase the overall effectiveness of trainings. Future training programs could use a strategic reflection process to emphasize skill development in the design and delivery of interpretation. Results derived from this empirical research provide a guideline for developing a training framework contextualized to the practice of interpretation in Japan.

RÉSUMÉ:Cet article explore, en examinant leurs croyances saillants, les facteurs qui déterminent si les personnes âgées participent aux Jeux des aînés et de l’activité physique durant les loisirs connexes. Nous avons mené 10 entrevues en profondeur avec des personnes âgées qui ont participé aux Jeux des aînés. Nous avons exploré trois types de croyance sous-tendus par le cadre théorique de comportement prévu: les avantages et les inconvénients (croyances comportementales), le soutien social et la pression (croyances normatives), et les facilitateurs et les obstacles (croyances de contrôle). Les répondants aux entrevues se sont trouvés à s’engager aux Jeux des aînés et à l’activité physique dans la mesure où ils ont associé divers avantages intangibles avec les Jeux et en ont valorisé la satisfaction psychologique. Ils considéraient leurs pairs et les familles comme soutenant et approuvant leur engagement, tout en reconnaissant les capacités physiques et les contraintes structurelles nécessaires pour se livrer aux Jeux et les activités connexes. Les résultats indiquent comment les croyances pertinentes peuvent être combinées avec des interventions pour encourager pour les personnes âgées des activités physiques des loisirs. ABSTRACT:This article explores the factors determining whether older adults engage in the Senior Games and related leisure-time physical activity through examining the adults’ salient beliefs. We conducted 10 in-depth interviews with older adults who have participated in the Senior Games. Underpinned by the planned behavior theory’s framework, we explored three types of beliefs: advantages and disadvantages (behavioral beliefs), social support and pressure (normative beliefs), and facilitators and impediments (control beliefs). Interview respondents were found to engage in the Senior Games and related physical activity to the extent that they associated various intangible advantages with the games and valued psychological satisfaction. They viewed their peers and families as supporting and approving of their engagement and recognized the physical capabilities required, and structural constraints necessary, to engage in the games and related activity. With these findings, pertinent beliefs can be combined with interventions designed to encourage leisure-time physical activities by older adults.

RÉSUMÉCet article explore, en examinant leurs croyances saillants, les facteurs qui déterminent si les personnes âgées participent aux Jeux des aînés et de l’activité physique durant les loisirs connexes. Nous avons mené 10 entrevues en profondeur avec des personnes âgées qui ont participé aux Jeux des aînés. Nous avons exploré trois types de croyance sous-tendus par le cadre théorique de comportement prévu: les avantages et les inconvénients (croyances comportementales), le soutien social et la pression (croyances normatives), et les facilitateurs et les obstacles (croyances de contrôle). Les répondants aux entrevues se sont trouvés à s’engager aux Jeux des aînés et à l’activité physique dans la mesure où ils ont associé divers avantages intangibles avec les Jeux et en ont valorisé la satisfaction psychologique. Ils considéraient leurs pairs et les familles comme soutenant et approuvant leur engagement, tout en reconnaissant les capacités physiques et les contraintes structurelles nécessaires pour se livrer aux Jeux et les activités connexes. Les résultats indiquent comment les croyances pertinentes peuvent être combinées avec des interventions pour encourager pour les personnes âgées des activités physiques des loisirs.

We have demonstrated that ribosomal protein S19 (RP S19) polymer, when crosslinked between Lys122 and Gln137 by activated coagulation factor XIII, acts as a C5a receptor (C5aR) antagonist/agonist. Based on experimental data obtained using RP S19 analog peptide and recombinant protein monomer, we suggested that L131DR, I134AGQVAAAN and K143KH moieties in the RP S19 C‐terminus act in, respectively, C5aR binding, penetration of the plasma membrane, and interaction with either an apoptosis-inducing molecule in neutrophils (delta lactoferrin) or a calcium channel-activating molecule (annexin A3) to induce the p38 MAPK pathway in macrophages. Recently, we observed RP S19 trimer in serum. To study the effects of this RP S19 trimer on C5aR, we prepared mutant RP S19 C‐terminal peptide (RP S19122-145) dimer and trimer, and examined their chemotactic activities and signal transduction pathways in human C5aR-overexpressing squamous cell carcinoma HSC-1 (HSC-1C5aR) cells using 24 trans-well chamber and western blotting assays, respectively. HSC-1C5aRcells were attracted by RP S19122-145dimer and vice versa by RP S19122-145trimer. The RP S19122-145dimer-induced attraction was competitively blocked by pre-treatment with RP S19122-145trimer. Moreover, RP S19122-145trimer-induced p38 MAPK phosphorylation was stronger than RP S19122-145dimer-induced p38 MAPK phosphorylation. RP S19122-145trimer appeared to act as a C5aR antagonist. The agonistic and antagonistic effects of RP S19122-145dimers and trimers were reflected by monocytic, THP-1-derived macrophage-like cells. Unlike the C5aR agonist C5a, which acts at the inflammation phase of acute inflammation, RP S19 trimer might act as a C5aR antagonist at the resolution phase.

This research attempted to identify the needs present in interpreter training in Japan. Interviews with trainers and interpreters, and questionnaires administered to interpreters were employed to explore (a) the skills and abilities necessary for successful interpreters and (b) the challenges that trainers may experience regarding interpretive training in Japan. The results showed both consistency and inconsistency in the perceptions of interpreters and trainers. Interpretive design, communication and public speaking, and risk management are core subjects recommended for future introductory-level training programs in Japan. Managerial and training skills are suggested for new types of training programs. Several challenges to interpreter training in Japan were also identified.

Graduates with strong foreign language skills (FLS) have an advantage when applying for a job. Underpinned by the theory of planned behavior, this research aimed at examining influences on the intentions of tourism majors in a Korean university to acquire FLS. A first survey was conducted with 256 students who had completed two or more semesters in the program. A second survey was administered with 134 students who had newly entered the program. Overall, the attitudes and subjective norms were found to be significant contributors, while behavioral control appeared to be less influential in regard to the students’ intentions to acquire FLS. Different interventions to influence the students’ intentions to acquire FLS should be considered based on the students’ enrollment periods. Effects can be larger on students within certain enrollment periods if interventions are targeted at normative beliefs, while effects may be larger on students who are new to the program if interventions highlight control beliefs.

Transplantation of rat hepatocytes into the syngeneic rat spleen results in the appearance of cytokeratin (CK)7 and CK19 positive biliary cells that form ductules. We examined whether hepatocytes are the origin of these biliary ductular cells.

Freezing behavior is thought to be a sign of fear in animals. We examined whether the freezing behavior during the elevated open-platform stress, which is a psychological stressor without painful stimulus, is modulated by serotonergic neurotransmission and would be a useful marker for screening anxiolytic and/or antidepressant. Male ICR mice (6 – 8-week-old) were individually placed on an elevated open-platform and the duration of freezing behavior of mouse was measured for 10 min. Fluoxetine and citalopram, selective serotonin (5-HT) reuptake inhibitors, markedly decreased the duration of freezing. Fenfluramine, a 5-HT releaser, and 8-OH-DPAT, a potent 5-HT1A-receptor agonist, also significantly decreased the duration of freezing. In contrast, the 5-HT-synthesis inhibitor p-chlorophenylalanine significantly increased the duration of freezing. Diazepam, a benzodiazepine anxiolytic, had no effect on the duration of freezing at doses having no effect on locomotor activity. Imipramine and clomipramine, tricyclic antidepressants, also did not affect the duration of freezing. Reboxetine, a selective noradrenaline reuptake inhibitor, significantly increased the duration of freezing. These results indicate that the activation of serotonergic neurotransmission attenuates the fear-related behavior in the elevated open-platform test, while the activation of noradrenergic neurotransmission increases the fear-related behavior. In addition, this test is convenient for assaying anxiolytic drugs that affect serotonergic neurotransmission.

To examine the usefulness of interleukin-18 (IL-18) in the treatment of osteosarcomas, the effect of IL-18 on the growth of Dunn osteosarcoma cells was investigated. Daily intraperitoneal (i.p.) injection of mouse recombinant IL-18 (2 μg/mouse) suppressed the growth of Dunn osteosarcoma cells transplanted subcutaneously (s.c.) into syngeneic C3H mice. This IL-18-induced suppression was not affected by simultaneous treatment with anti-asialo GM1 serum, which inactivates natural killer (NK) cells. However, IL-18 failed to suppress the growth of Dunn osteosarcoma cells transplanted into BALB/c-nude mice devoid of T lymphocytes or C3H-gld/gld mice deficient in functional Fas ligand (FasL). IL-18 also failed to suppress the growth of Dunn osteosarcoma cells in vitro, although expression of IL-18 receptor mRNA and MyD88 mRNA as well as Fas mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). On the other hand, antimouse Fas antibody showed cytotoxicity against Dunn osteosarcoma cells in a dose-dependent manner in vitro. In addition, treatment of C3H mice with IL-18 enhanced the cytotoxic activity of CD8+ T lymphocytes against Dunn osteosarcoma cells. These results indicate that IL-18 inhibits the growth of Dunn osteosarcoma cells in vivo by enhancing the cytotoxic activity of CD8+ T lymphocytes through the FasL-Fas system.

Transplantation of rat hepatocytes into the syngeneic rat spleen results in the appearance of cytokeration (CK)-19-positive biliary cells that form ductules. The exact origin of CK-19-positive cells is not known and the possibility that they are derived from biliary cells or precursors of oval cells in transplanted hepatocyte preparations has been raised. In the present study, we found that the number of CK-19-positive biliary cells increased rapidly after transplantation of hepatocytes, reached the maximum at 4 weeks, and then gradually decreased. However, a Ki-67 labeling index of CK-19-positive biliary cells was low and showed no significant changes throughout the experimental period. In addition, no or few CK-19-positive cells appeared in the spleen after transplantation of nonparenchymal liver cells enriched with biliary cells. These results showed that biliary cells were not the source of CK-19-positive cells in the spleen. Impairment of precursors of oval cells in the liver by administration of 4,4′-diaminodiphenylmethane 24 h before transplantation of hepatocytes did not prevent the appearance of CK-19-positive biliary cells in the spleen. Moreover, transplantation of nonparenchymal cells carrying an increased number of oval cells by means of treatment with 2-acetylaminofluorene and partial hepatectomy resulted in no appearance of CK-19-positive biliary cells in the spleen. These results ruled out oval cells as the origin of CK-19-positive biliary cells in the spleen. Because CK-19-positive biliary cells appeared in the spleen only when hepatocyte fractions were transplanted, we suggest transdifferentiation of heptocytes may be the mechanism by which CK-19-positive biliary cells are generated.

The release of cytochrome c from mitochondria, which is regulated by Bcl-2 family members and is considered to take place through voltage-dependent anion channels (VDACs) on the outer membranes of mitochondria, results in activation of effector caspases, such as caspase-3, which induce apoptosis. We studied the involvement of the mitochondrial apoptosis pathway in uterine epithelial apoptosis. Estradiol-17β pellets were implanted into ovariectomized mice and removed 4 days later (Day 0). The apoptotic index (percentage of apoptotic cells) of the luminal epithelium increased markedly, peaking on Day 2, whereas that of the glandular epithelium increased much less. Expression of VDAC1, 2, and 3 mRNAs increased in the luminal epithelium in correlation with the apoptotic index of the luminal epithelium. No increases in VDAC1, 2, and 3 mRNA levels were observed in the stroma or muscle, where no apoptosis occurs. VDAC1 protein levels in the uterus also correlated well with the apoptotic index of the luminal epithelium. In addition, the apoptotic index showed good correlation with the release of cytochrome c from mitochondria, activation of caspase-3, which was immunohistochemically detected only in the epithelium, and the mRNA and protein ratios of Bax:Bcl-2 and Bax:Bcl-X in the uterus. The present results suggest that the release of cytochrome c from mitochondria, which is regulated by Bcl-2 family members, plays a role in uterine epithelial apoptosis after estrogen deprivation. The increase in VDAC expression may facilitate the release of cytochrome c during apoptosis.

It has recently been proposed that gastrointestinal stromal tumors (GISTs) originate from stem cells that differentiate toward a phenotype of interstitial cells of Cajal (ICCs). Nestin is a newly identified intermediate filament protein, and is predominantly expressed in immature cells, such as neuroectodermal stem cells and skeletal muscle progenitor cells, and tumors originating from these cells. In this study, we examined, using immunohistochemistry, the nestin expression in GISTs and ICCs to clarify the origin of GISTs. Strong immunoreactivity for nestin was observed in all 18 GISTs, and its expression was confirmed by Western blot and Northern blot analyses. In contrast, three leiomyomas and a schwannoma that developed in the gastrointestinal tract showed no apparent immunoreactivity for nestin. Among 17 mesenchymal tumors (seven leiomyosarcomas, five malignant peripheral nerve sheath tumors, and five fibrosarcomas) that occurred in sites other than the gastrointestinal tract, only two malignant peripheral nerve sheath tumors were moderately immunoreactive for nestin. Furthermore, with fluorescence double immunostaining of the normal small intestine, nestin expression was demonstrated in ICCs. These results show that nestin may be a useful marker for diagnosis of GISTs, and support the current hypothesis that GISTs are tumors of stem cells that differentiate toward an ICC phenotype.

Osteoprotegerin (OPG) and osteoclast differentiation factor (ODF) are crucial regulators of osteoclastogenesis. To determine the biological role of interleukin (IL)-18 produced by stromal/osteoblastic cells in osteoclastogenesis, we examined the effects of IL-18 on the OPG and ODF mRNA levels in these cells. When bone marrow stromal ST2 cells, osteoblastic MC3T3-E1 cells, and mouse calvarial osteoblasts were stimulated with IL-18, the expression of OPG mRNA, but not ODF mRNA, was transiently increased, its expression reaching a maximal level at 3 h after the beginning of the culture. In accordance with this observation, all these cells expressed the mRNAs of two IL-18 receptor components and MyD88, an adapter molecule involved in IL-18 signaling. Moreover, in these cells, mitogen-activated protein kinase was phosphorylated after stimulation with IL-18. These results suggest that stromal/osteoblastic cells are IL-18-responsive cells and that IL-18 may inhibit osteoclastogenesis by up-regulating OPG expression, without stimulation of ODF production, in stromal/osteoblastic cells.

The effects of neonatal hypothyroidism on the gene expression of progesterone receptors (PRs) in the 8-day-old female rat brain were examined. The levels of PR (‘A’+‘B’)-, PR (‘B’)-, estrogen receptor (ER)- and β-actin mRNAs in the cerebral cortex and hypothalamus-preoptic area of propylthiouracil-treated rats (PTU group) or untreated rats (control group) were analyzed using a quantitative reverse transcription-polymerase chain reaction-Southern blotting assay. When the levels of PR mRNAs were calibrated by respective levels of β-actin mRNA, the levels of PR (‘A’ + ‘B’)- and PR (‘B’) mRNAs in the cerebral cortex of the PTU group were markedly less than those in the control group with no significant changes in the levels of PR messages in the hypothalamus-preoptic area of both groups. No significant difference in the calibrated levels of ER mRNA between both groups was found in these tissues. These results on the PR-, and ER mRNAs were essentially similar to those on the levels of PR- and ER proteins previously reported suggesting that thyroid hormones affect the transcriptional machinery of PR in the developing brain, in a region-specific manner.

The structural damage to and leakage of internal substances from Saccharomyces cerevisiae 0–39 cells induced by hydrostatic pressure were investigated. By scanning electron microscopy, yeast cells treated at room temperature with pressuresbellw 400 MPa for 10 min showed a slight alteration in outer shape. Transmission electron microscopy, however, showed that the inner structure of the cell began to be affected, especially the nuclear membrane, when treated with hydrostatic pressure around 100 MPa at room temperature for 10 min; at more than 400–600 MPa, further alterations appeared in the mitochondria and cytoplasm. Furthermore, when high pressure treatment was carried out at — 20° C, the inner structure of the cells was severely damaged even at 200 MPa, and almost all of the nuclear membrane disappeared, although the fluorescent nucleus in the cytoplasm was visible by 4,6-diamidino-2-phenylindole (DAPI) staining. The structural damage of pressure-treated cells was accompanied by the leakage of internal substances. The efflux of UV-absorbing substances including amino acid pools, peptides, and metal ions increased with increase in pressure up to 600 MPa. In particular, amounts of individual metal ion release varied with the magnitude of hydrostatic pressures over 300 MPa, which suggests that the ions can be removed from the yeast cells separately by hydrostatic pressure treatment.

Three cDNA clones (cat1, cat2, cat3) for catalase (EC 1.11.1.6) were isolated from a cDNA library of pumpkin (Cucurbita sp.) cotyledons. In northern blotting using the cDNA-specific probe, the cat1 mRNA levels were high in seeds and early seedlings of pumpkin. The expression pattern of cat1 was similar to that of malate synthase, a characteristic enzyme of glyoxysomes. These data suggest that cat1 might encode a catalase associated with glyoxysomal functions. Furthermore, immunocytochemical analysis using cat1-specific anti-peptide antibody directly showed that cat1 encoding catalase is located in glyoxysomes. The cat2 mRNA was present at high levels in green cotyledons, mature leaf, stem and green hypocotyl of light-grown pumpkin plant, and correlated with chlorophyll content in the tissues. The tissue-specific expression of cat2 had a strong resemblance to that of glycolate oxidase, a characteristic enzyme of leaf peroxisomes. During germination of pumpkin seeds, cat2 mRNA levels increased in response to light, although the increase in cat2 mRNA by light was less than that of glycolate oxidase. cat3 mRNA was abundant in green cotyledons, etiolated cotyledons, green hypocotyl and root, but not in young leaf. cat3 mRNA expression was not dependent on light, but was constitutive in mature tissues. Interestingly, cat1 mRNA levels increased during senescence of pumpkin cotyledons, whereas cat2 and cat3 mRNAs disappeared during senescence, suggesting that cat1 encoding catalase may be involved in the senescence process. Thus, in pumpkin, three catalase genes are differentially regulated and may exhibit different functions.

Fecal bile acid and fecal fat were determined in 18 normal subjects and 22 patients with chronic pancreatitis, and the relation of fecal bile acid excretion to exocrine pancreatic dysfunction was studied. In chronic pancreatitis fecal bile acid was approximately three times that of control subjects, and large amounts of primary bile acid were detected. A significant correlation between fecal bile acid excretion and bicarbonate secreted from the pancreas was found. This evidence of bile acid malabsorption was not observed until bicarbonate output was ∼0.05 mEq/h/kg. A slight correlation between fecal bile acid and absorption rates of fat was demonstrated. These results suggest that bile acid malabsorption observed in chronic pancreatitis is related to an impairment of pancreatic bicarbonate secretion.

A determination of caloric consumption based on a dietary survey table, fat and cholesterol intake, and analyses of fecal fatty acids and neutral sterols, and bile acid analysis (gas chromatographic method) were conducted on 33 subjects (including 17 patients with chronic pancreatitis and 16 normal controls). The factors related to hypocholesterolemia in chronic pancreatitis (CP) patients were investigated and the following conclusions were obtained: (1) The total caloric intake and fat consumption by the CP patients were significantly lower with the exception of cholesterol consumption. (2) Significant increases were noted in fecals fat, neutral sterols, and bile acid excretion by the CP patients. (3) A significant positive correlation was noted between the total cholesterol and body mass index (BMI), reaffirming that the cholesterol level can be used as an indicator of nutritional status. (4) A significant negative correlation was noted between the serum total cholesterol and fecal bile acid excretion. These findings indicate that CP patients suffer from neutral sterol malabsorption, in addition to dietary fat maldigestion and bile acid malabsorption. Furthermore, bile acid malabsorption is cited as a factor in the development of hypocholesterolemia in CP patients.

Conclusion: Pancreatic endocrine capacities are remarkably disturbed in patients with pancreatic diabetes owing to calcific pancreatitis as opposed to those owing to noncalcific pancreatitis. Insulin secretion in calcific pancreatitis resembled, that in insulin-dependent diabetes mellitus (IDDM), whereas insulin secretion in noncalcific pancreatitis resembled that in non-IDDM (NIDDM). The involvements of acinar cell and ductal cell function closely correlate with endocrine function (insulin and glucagon secretions) in chronic pancreatitis (pancreatic diabetes). Background: We sought to clarify the differences of pancreatic endocrine function between pancreatic diabetes and primary diabetes, and to verify the correlations between pancreatic exocrine and endocrine dysfunction in patients with chronic pancreatitis. Methods: Urinary C-peptide (CPR) excretion and fasting plasma glucagon levels in patients with pancreatic diabetes owing to calcific pancreatitis (19 cases) and owing to noncalcific pancreatitis (14 cases) were studied in comparison with those in patients with insulin-dependent diabetes mellitus (IDDM, 23 cases), noninsulin-dependent diabetes (NIDDM, 18 cases), and in healthy controls (11 cases). In addition, pancreatic exocrine function was investigated in patients with chronic pancreatitis (calcific and noncalcific) and in healthy controls. The correlation between pancreatic exocrine and endocrine function was studied. Results: The urinary CPR excetion in controls was 94.9±20.5 μg/d. The urinary CPR excretion in calcific pancreatitis was 12.8±7.4 μg/d and it resembled that in IDDM (9.4±5.8 μg/d). The urinary CPR excretion in noncalcific pancreatitis was 41.5±30.1 μg/d, being similar to that in NIDDM (49.3±21.0 μg/d).

The study was conducted on five healthy subjects and six patients with calcifying pancreatitis (CP) and steatorrhea. Following overnight fasting, one tube each was placed in the stomach and the upper part of the small intestine, respectively. Through the gastric tube, a test meal that included 30 g of fat (total calories, 625 kcal, 500 mL) was infused over a span of 30 min. Every 30 min (up to 150 min), fluid samples in the upper small intestine were collected and chilled, and the amylase, trypsin, and lipase levels were determined. In addition, in the case of the CP patients, a high-potency pancreatin preparation was infused into the stomach together with the test meal. In order to determine the plasma CCK level, blood sample were collected before test meal infusion and at 10, 20, 30, 45, 60, 90, 120, and 150 min subsequent to infusion. The plasma CCK was extracted using a Sep-Pak C-18 cartridge and analyzed with radioimmunoassay using an OAL-656 antibody. The result was converted to the CCK-8 level and expressed in pg/mL. The enzyme activities in the upper small intestine of the CP patients after test meal administration amounted to 22.8 (amylase), 10.8 (trypsin), and 16.9% (lipase) compared with the corresponding figures for the normal subjects. Following administration of a high-potency pancreatin in patients with CP, enzyme activities in the upper small intestine increased to 132.2 (amylase), 38.7 (trypsin), and 45.3% (lipase) compared with levels in the normal subjects. However, the healthy subjects and the CP patients, both with and without treatment with supplementary exogenous enzymes, all exhibited similar profiles in the plasma CCK response to stimuli. Based on these findings, we concluded that a negative feedback mechanism does not exist between the tryptic activity of the upper small intestine and the CCK secretory response in patients with chronic pancreatitis.

Conclusions: Steatorrhea was almost completely stopped and malabsorption of neutral sterols and shortchain fatty acids was reduced by treatment of high-lipase pancreatin in Japanese patients with pancreatic insufficiency whose dietary fat consumption is low. Methods: Fifteen patients with chronic pancreatitis complicated by steatorrhea who consumed an average of 48 g of dietary fats a day were selected as subjects and given 3 g of high-lipase pancreatin (lipase, 379,800 USP U/g), at each meal (total daily dose is 9 g) for a mean duration of 28.5 d. Fecal output and fecal fat neutral sterol, bile acid, and short-chain fatty acid excretion were determined before and after the course of pancreatin therapy. Results: Pancreatin administration resulted in significant reductions (P<0.01) in fecal output (from 243.2 to 149.1 g), excretion of fecal fat, (from 12.3 to 3.9 g), animal sterols (from 816.3 to 604.6 mg), and shortchain fatty acids (from 52.6 to 18.5 mM). In contrast, no marked changes were recorded in fecal excretion of ß-sitosterol (a plant sterol), bile acids, or the hydroxy fatty acid fraction. Fecal fat and short-chain fatty-acid excretion showed strong correlations with fecal output.

Conclusion: Visual observation of feces, considering fecal output, is considered to be an excellent method of detection of steatorrhea when judged by experienced doctors. Methods: Feces from 192 patients with untreated chronic pancreatitis or under pancreatic enzyme therapy were investigated. Feces were collected for three consecutive days and homogenized with water. Fecal samples were freeze-dried and analyzed for fatty acids by gas chromatography (GLC). The quantity of fat was calculated from the amount of fatty acid to obtain daily fecal fat excretion. Comparison of GLC method with van de Kamer method gave a significant (p<0.01) positive correlation with correlation coefficient of 0.916 (n=38). Steatorrhea was defined as fecal fat excretion exceeding 5 g/d. Mild steatorrhea was defined as 5–10 g/d, and severe steatorrhea as more than 10 g/d. Results: Three visual identification items were used to consider fecal output exceeding 200 g/d: fecal fat concentration exceeding 4%, appearance, and odor. The results were compared with the results from GLC method. Detection of steatorrhea by means of visual properties was the most accurate, and correlation coefficient was 0.843 (p<0.01) by Spearman’s rank correlation test. This detection method was also significantly effective for differentiation of normal stool from mild and severe steatorrhea. The sensitivity and specificity were 89.3 and 91.1%, respectively, indicating a favorable result.

To clarify the involvement of actin in the formation of the yeast cell wall, reverting protoplasts of Schizosaccharomyces pombe were used as a simple model system. Actin of reverting protoplasts was labeled with rhodamine-conjugated phalloidin and observed by conventional fluorescence microscopy and laser scanning confocal microscopy. A close spatial as well as temporal relationship between actin and cell wall formation was observed in protoplast reversion. That is, the site of actin‘dots’ in the reverting protoplasts coincided with the site of new wall formation and the timing of rearrangement of actin coincided with the initiation of cell wall formation and with the timing of cell wall expansion. Treatment of reverting protoplasts with cytochalasin D (CD) further clarified the close relationship between actin and cell wall organization. The effect of CD was dose dependent. A high dose of CD caused the absence of actin as well as the complete inhibition of cell wall formation. A low dose of CD caused weakly stained unlocalized actin, which induced grossly aberrant cell wall deposition as well as substantial changes in the morphology of the reverting protoplasts. These results demonstrated that actin is associated with initiation of cell wall formation, the proper deposition of cell wall materials, and maintaining the normal morphology of reverting protoplasts. Scanning electron microscopy revealed the presence of a fibrillar net structure on the surface of non-treated control reverting protoplasts. However, the absence of a fibrillar network on the surface of reverting protoplasts was observed in the presence of a high concentration of CD. Lack of localization of microfibrils as well as poor development of the fibrillar network were also observed in the presence of a low concentration of CD. Recovery experiments confirmed the close relationship between actin and cell wall formation.

Fecal fat excretion, fecal mass, fecal fat concentration, and the coefficient of fat absorption were evaluated in 31 normal Japanese subjects with a mean fat consumption of 61.8 g and compared with the values in 43 Japanese patients with chronic pancreatitis (CP) with a fat consumption of 40.2g. Fecal fatty acids were analyzed by the gas Chromatographic method. Fecal fat excretion by normal individuals was 1.7±1.0 (Mean±SD) g/day (range, 0.4–4.9 g/day). Steatorrhea was therefore defined as fecal fat excretion that exceeded 5.0 g/day. The patients with CP were divided into three groups: non-steatorrhea (fecal fat «5.0g/day), chemical steatorrhea (fecal fat =5 g/day and no appearance of fatty stool), and manifest steatorrhea (evaluated from the appearance of fatty stool). In addition, we investigated the correlation between fecal fat excretion and pancreatic exocrine function, obtained by the pancreozymin-secretin (or secretin) test in 24 controls and 30 CP patients. Fecal fat excretion by CP patients was 9.1±8.8 g/day, which was significantly higher (P<0.01) than that of controls. There were 28 (65%) CP patients with steatorrhea. Of these, 15 (35%) showed chemical steatorrhea and 13 (30%) manifest steatorrhea. In CP patients, the fecal mass (250.5±133.6 g vs control, 125.6±52.5 g), fecal fat concentration (3.40±2.16g% vs control, 1.48±0.89g%), and coefficient of fat absorption (77.3±20.2% vs control, 97.2±1.7%) all showed significant differences from the controls. Of these three indices—fecal mass, fecal fat concentration, and coefficient of fat absorption—fecal fat concentration was the most useful for identifying pancreatic steatorrhea. In addition, there were significantly different correlations between fecal fat excretion and pancreatic exocrine function in patients' group and control group. Fecal fat excretion was clearly correlated with amylase output. These findings led to the conclusion that steatorrhea indeed exists among Japanese CP patients who consume small amounts of dietary fat; and that fecal fat excretion is closely correlated with pancreatic exocrine dysfunction.

Summary: The study was conducted on five healthy subjects and six patients with calcifying pancreatitis (CP) and steatorrhea. Following overnight fasting, one tube each was placed in the stomach and the upper part of the small intestine, respectively. Through the gastric tube, a test meal that included 30 g of fat (total calories, 625 kcal, 500 mL) was infused over a span of 30 min. Every 30 min (up to 150 min), fluid samples in the upper small intestine were collected and chilled, and the amylase, trypsin, and lipase levels were determined. In addition, in the case of the CP patients, a high-potency pancreatin preparation was infused into the stomach together with the test meal. In order to determine the plasma CCK level, blood sample were collected before test meal infusion and at 10, 20, 30, 45, 60, 90, 120, and 150 min subsequent to infusion. The plasma CCK was extracted using a Sep-Pak C-18 cartridge and analyzed with radioimmunoassay using an OAL-656 antibody. The result was converted to the CCK-8 level and expressed in pg/mL. The enzyme activities in the upper small intestine of the CP patients after test meal administration amounted to 22.8 (amylase), 10.8 (trypsin), and 16.9% (lipase) compared with the corresponding figures for the normal subjects. Following administration of a high-potency pancreatin in patients with CP, enzyme activities in the upper small intestine increased to 132.2 (amylase), 38.7 (trypsin), and 45.3% (lipase) compared with levels in the normal subjects. However, the healthy subjects and the CP patients, both with and without treatment with supplementary exogenous enzymes, all exhibited similar profiles in the plasma CCK response to stimuli. Based on these findings, we concluded that a negative feedback mechanism does not exist between the tryptic activity of the upper small intestine and the CCK secretory response in patients with chronic pancreatitis.

To reveal the ultrastructure of regenerating protoplasts of Schizosaccharomyces pombe, samples were treated with ruthenium tetroxide (RuO4) after conventional fixation and then observed with a transmission electron microscope. Ruthenium tetroxide treatment was very effective for observing the ultrastructure. The treatment provides electron-dense and clear images of glucan fibrils developing from the protoplast surface. The filasomes, Golgi apparatus, and secretory vesicles with high-electron density were also observed in the cytoplasm. Ruthenium tetroxide treatment allows the interior and exterior ultrastructure of the cell to be observed clearly at the same time.

The ultrastructure of regenerating cell wall in Schizosaccharomyces pombe protoplasts was studied with a high resolution, low voltage scanning electron microscope (LVSEM). In contrast to tke transmission electron microscopy, the LVSEM images give three-dimensional information on the cell wall regeneration in yeast protoplasts. We found that, after only a few minutes of incubation, the protoplasts began to show protuberances in a unipolar manner, and a fibrilar network was formed asymmetrically which covered the whole surface of the protoplasts after 5 hr. The network consisted of microtibrils about 8 to 10 nm wide, forming flat and wavy bundles of various widths and lengths, up to about 200 nm wide and 1 μm long, mainly made of yeast glucan. Free ends of microfibrils were seldom found. Interfibrillar spaces were progressively filled with granular particles and finally the complete cell wall was formed after 12 hr. The fibrillar network was destroyed by the digestion with β(1→3)-glucanase. When protoplasts were regenerating in the presence of aculeacin A, the fibrillar networks were not formed, resulting in incomplete cell wall formation. These observations suggest that β9-glucan is the main component of the microfibrils and that it plays an important role in the formation of the cell wall in S. pombe.

Prolamellar bodies (PLBs) were isolated from etioplasts of Cucurbita moschata Durch var.melonaeformis Makino, and their fractured planes were observed at high magnification with a high-resolution scanning electron microscope (SEM) to elucidate the paracrystalline structure of the PLB. Thus, we obtained a standard method of preparation of biological specimens. A good preparation should provide a high quality SEM image which will reveal fine structures of the specimens even at high magnification. In this experiment, the specimens were fixed with osmium tetroxide, conductlvely stained with uranyl acetate, dehydrated with alcohol series, rapidly frozen in liquid Freon, and freeze-fractured in liquid nitrogen. It was proved that the specimens correctly retained their fine structures, and this suggested that very thin, high quality platinum-carbon coating (2 nm in thickness) by rotary evaporation with the electron beam gun provide the best high-resolution contrast for the fractured specimens. Through these procedures, the three-dimensional structure of the PLBS could be observed precisely at high magnification using a high-resolution SEM.

The time course of BMY-28864-dependent morphological changes in Candida albicans A9540 was studied by electron microscopy. Scanning electron microscopy revealed that BMY-28864 often induced cell surface deformations such as abnormal swelling and bulging around budding sites and bud scars. The cell membrane damage was visualized by freeze-fracturing technique as deep pit-like invaginations. Transmission electron microscopy with thin-sectioned specimens also demonstrated that BMY-28864 induced cell membrane invaginations together with cell membrane detachment from the cell wall, nuclear membrane fragmentation and mitochondrial aberration. Statistical sequence analysis of the prominent BMY-28864-dependent morphological changes of the yeast cells led to the conclusion that BMY-28864 first attacked the cell membrane and then caused disintegration of other intracytoplasmic organelles, resulting in the lethal effect.

The refined field emission SEM, S-900 LV which gives better resolution especially at low voltages below 5 kV was developed for ultrahigh resolution scanning electron microscopy. A visualization test at × 300,000 was made using a gold-evaporated magnetic tape, and the resolution was found to be about 1 nm at 2.5 kV. The ultrastructure of the cell wall, especially the reverting glucan network, from the protoplast of Schizosaccharomyces pombe was studied using this improved ultrahigh-resolution low-voltage SEM (UHR-LVSEM). The results with uncoated reverting protoplasts observed with this microscope revealed that the network was originally formed as secreted particles scattered on the protoplast surface and these were subsequently stretched to microfibrils about 2 nm thick. The microfibrils were twisted around each other and joined together so that they developed into 8-nm-thick fibrils, forming a ribbon-shaped network of glucans about 16-nm-thick which covered the entire protoplast surface. The UHR-LVSEM images of reverting protoplasts treated with glucanase confirmed that the particles scattered on the protoplast surface in the initial stage of regeneration were glucan in nature.

Uncoated yeast cells prepared by the freeze-substltution fixation and cryofracturing method were studied witb an “In-lens” type field emission scanning electron microscope (FESEM) at low voltage (0.8-3 kV). The micrographs were compared with those obtained with a conventional FESEM and coated specimens, as well as witb transmission electron micrographs of ultrathin-sectioned yeast cells. The images of uncoated fractured cells taken at 1.2-1.5 kV seem to show the highest fidelity and thus they presented their threedimensional structures more dynamically. The fine filaments produced on the outermost surface of the cell wall were also clearly observed witb a low-voltage scanning electron microscope. The filaments of a similar specimen taken at 20 kV after coating with Pt-C 2 nm in thickness, however, showed an enlarged and intertwined structure because of a masking and decoration effect due to coating. In addition to several intracellular organelles, free ribosomes in the cytoplasm and Intramembranous particles of the cell membrane were revealed by the “In-lens” FESEM and these images correspond well to those obtained with uitrathin sectioning and freeze-fracturing replicas in the transmission electron microscope.

Until recently, a mixture of backscattered electron signal and secondary electron (SE) signal of gold-labeled specimens has been used for the concurrent visualization of the surface topography and localization of gold particles in biological specimens by scanning electron microscopy (SEM). Using a new ultra-high resolution field emission scanning electron microscope (UHR-FESEM), S-900 LV, however, it is possible to observe concurrently the surface structure and gold particles using only the SE mode at a low voltage (1–3 kV). We confirmed the location of α-galactomannan on the protoplast membrane of Schizosaccharomyces pombe by this method and determined the process of formation of α-galactomannan in the regeneration of cell wall from S. pombe protoplasts utilizing gold-labeled lectin. α-Galactomannan formed in the interfibrillar network of glucan in the reverting protoplast at 5 hr and progressively filled interfibrillar spaces. Observation at low voltage with SE is a promising approach to determine the location of specific molecules on a specimen surface using colloidal gold.

Fungi collectively known as Eurotium (Aspergillus) restrictus group form one of the predominant groups causing the biodeterioration of cultural assets. Six species of E. (A.) restrictus including 10 type strains and E. (A.) isolate No. 3 were examined by scanning electron microscopy (SEM). As surface ornamentation of the conidial spores from these 11 strains was found to differ each other, SEM observation of the spores serves as a simple and useful method to identify these species. Recently, E. (A.) isolate No. 3 was successfully cultured and isolated under high osmotic pressure, and was regarded as a genuine fungus causing foxing. When the foxed area of a sheet of paper taken from an old document was kept under high humidity, fungal colonies appeared. The conidial spores were found to be identical to those of E. (A.) isolate No. 3. Thus, the microflora of the biodeterioration process were identifiable by direct SEM examination.