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Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA) and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species.

In the past several years, the liberalization of the electricity supply, the increase in variability of electric appliances and their use, and the need to respond to the electricity demand in real time has made electricity demand forecasting a challenge. To this challenge, many solutions are being proposed. The electricity demand involves many sources such as economic activities, household need and weather sources. All of these sources make electricity demand forecasting difficult. To forecast the electricity demand, some proposed parametric methods that integrate main variables that are sources of electricity demand. Others proposed a non parametric method such as pattern recognition methods. In this paper, we propose to take only the past electricity consumption information embedded in a functional vector autoregressive state space model to forecast the future electricity demand. The model we proposed aims to be applied at some aggregation level between regional and nation-wide grids. To estimate the parameters of this model, we use likelihood maximization, spline smoothing, functional principal components analysis and Kalman filtering. Through numerical experiments on real datasets, both from supplier Enercoop and from the Transmission System Operator of the French nation-wide grid, we show the appropriateness of the approach.

PACAP (pituitary adenylate cyclase-activating polypeptide) is a member of the VIP/secretin/glucagon family, which includes the ligands of class II G-protein coupled receptors. Since the recognition of PACAP by the receptor may involve the binding of PACAP to membranes, its membrane-bound structure should be important. We have carried out structural analysis of uniformly 13C,15N labeled PACAP27 and its C-terminal truncated form PACAP(1-21)NH2 (PACAP21) bound to membranes with high resolution solid-state NMR. Phosphatidylcholine bilayers and phosphatidylcholine/phosphatidylglycerol bilayers were used for PACAP27 and PACAP21, respectively. Most backbone signals were assigned for PACAP27 and PACAP21. TALOS analysis revealed that both peptides take on extended conformations on the membranes. Dilution of PACAP21 did not change the conformation of the major part. Selective polarization transfer experiment confirmed that PACAP27 is interacting with the membranes. It was concluded that the interaction of PACAP with the membrane surface causes their extended conformation. PACAP27 is reported to take an alpha-helical conformation in dodecylphosphocholine micelles and membrane-binding peptides usually take similar conformations in micelles and in membranes. Therefore, the property of PACAP27 changing its conformation in response to its environment is unique. Its conformational flexibility may be associated with its wide variety of functions.

A canine experimental model for the anomalous arrangement of the pancreaticobiliary ducts (APBD) was made to investigate the effects of bile acids on carcinogenesis. Seven adult mongrel dogs underwent dorsal pancreatico-cholecystostomy to serve as a functional model for APBD, and six dogs underwent the same procedure with the pancreatic duct ligated as a control group. Bile from the gallbladder was taken 14 months after surgery for bile acid analysis by HPLC. DNA strand breaks in HeLa cells induced by the bile were also investigated in situ by nick translation method. As a result, the fraction of cholic acid tended to be lower, and that of deoxycholic acid slightly higher in APBD-dogs (N.S.). The ursodeoxycholic acid percentage in APBD-dogs significantly decreased compared with that in the control and normal dogs (p < 0.05). Extremely high frequency of DNA strand breaks was shown in only two out of seven APBD-dogs. In those two dogs, the cholic acid percentage decreased and that of deoxycholic acid increased extremely. These findings suggest that the alteration of the bile composition in APBD caused frequent DNA strand breaks and repair which might lead to gene mutation and biliary tract carcinoma.

Anomalous arrangement of the pancreaticobiliary ducts is a congenital condition which predisposes the affected person to biliary tract carcinoma. We developed an experimental dog model of anomalous arrangement of the pancreaticobiliary ducts to investigate the mechanism of carcinogenesis in this condition. We used this model to analyze point mutations in the c-Ki-ras gene, and to assess the expression of mutant p53 protein in the gallbladder mucosa. The histopathological appearance of the gallbladder mucosa was also examined. Glandular structures were seen in four of seven (57%) gallbladders examined 14 months after the surgical creation of an anastomosis between the gallbladder and the pancreatic duct. Goblet cells were seen in two of seven gallbladders (29%). However, dot-blot hybridization and immunohistochemical study did not reveal any mutations in the c-Ki-ras gene, or any over-expression of the p53 protein in the specimens. These results show that the gallbladder mucosa is damaged by refluxing pancreatic juice in this dog model of anomalous arrangement of the pancreaticobiliary ducts, but that severe damage may be necessary to induce mutations in the c-Ki-ras proto-oncogene, or in the p53 gene.

We report herein the case of a 76-year-old man with intractable dumping syndrome which had manifested itself 3 years after he underwent a gastric resection and Billroth I reconstruction for a gastric ulcer. Despite aggressive medical therapy by the time of admission, he had suffered from disabling dumping symptoms for 9 years. In an attempt to relieve these symptoms, a 15-cm segment of the jejunum was placed isoperistaltically between the lesser curvature of the remnant stomach and the duodenum, and a selective vagotomy was performed. The dumping symptoms that he had experienced preoperatively completely disappeared after the revisory surgery. Postoperatively, an upper gastrointestinal series demonstrated a larger gastric pouch and slower gastric passage into the small intestine than what was seen preoperatively, while gastric emptying studies using the acetaminophen method also showed normal patterns in both the early and late postoperative phases. Thus, we consider that this surgical procedure is a simple and effective way to inhibit rapid gastric emptying and to slow intestinal transit in the treatment of dumping syndrome.

We have developed a new ATP assay for hepatocytes which is simple, rapid and highly sensitive. ATP determination using luciferin-luciferase was performed on hepatocytes separated by perfusion of the liver with collagenase. There was a close correlation between the ATP content of hepatocytes and viable cell numbers. The ATP content of dead cells which were determined by trypan blue dye exclusion test had less than one per cent of levels of viable cells. ATP contents of isolated hepatocytes in Minimum Essential Medium was 6.8 +/- 0.6 x 10(-15) mol/cell at 2 hours after excision of the liver and showed no significant difference compared with that determined at 6 hours. This method was performed to evaluate changes in the cellular ATP content of hepatocytes after partial hepatectomy in rats and transcatheter portal embolization (TPE) in dogs. The ATP content in rat hepatocytes showed a remarkable increase after hepatectomy, with a peak value of 19.1 +/- 1.7 x 10(-15) mol/cell at 24 hours post-surgery. On the other hand, marked atrophy in the embolized lobes and compensatory hypertrophy in the non-embolized lobes were found following TPE in dogs. Cellular ATP content in the non-embolized lobes showed its highest level of 8.7 +/- 2.9 x 10(-15) mol/cell on the third day after TPE, but in the embolized lobes decreased immediately after TPE with significant differences compared with the non-embolized lobes (p < 0.05). Our method may also be applicable to the evaluation of other adenosine phosphate by use of converting enzymes for ATP.(ABSTRACT TRUNCATED AT 250 WORDS)

Malignant rhabdoid tumor is initially differentiated from renal tumors and characterized by extremely rapid growth. We report a newborn case of extrarenal malignant rhabdoid tumor and review the literature.

Urethral prolapse is rarely reported in the pediatric literature. We report a 4 year old female with urethral prolapse and discuss the etiology and the treatments in children documented in the literature.

An ATP/ubiquitin-dependent proteasome complex with an apparent sedimentation coefficient of 26S was purified from rat liver to near homogeneity by an improved method based on procedures reported previously. Two electrophoretically distinct forms of the 26S complex, named 26S alpha and 26S beta, with very similar subunit compositions were found not only in purified preparations but also in crude extracts, indicating that the 26S proteasome is present as two isoforms. The 26S proteasome was shown to degrade multi-ubiquitinated, but not unmodified, lysozymes in an ATP-dependent fashion, to have ATPase activity supplying energy for proteolysis, and to contain isopeptidase activity to generate free ubiquitin Mg2+/ATP-dependently. The 26S proteasome also catalyzed the ATP-independent hydrolyses of three types of fluorogenic peptides with basic, neutral, and acidic amino acids at their cleavage sites, respectively. These peptides are also good substrates for the 20S proteasome, but their degradation by the free 20S proteasome and by its assembled form in the 26S complex differ markedly, suggesting a functional difference between the two forms of proteasomes. Electrophoretic and immunochemical analyses showed that the large 26S complex was composed grossly of two different structures: a core 20S proteasome with multicatalytic proteinase functions and an associated part possibly with a regulatory role. These two structures both consisted of multiple polypeptides with molecular masses of 21-31 and 35-110 kDa, respectively. The subunit multiplicity of the rat 26S proteasome closely resembled that of the human counterpart, showing only minor species-specific differences in certain components. The assembly of this multi-component complex was found not to involve a sulfhydryl bond. Electrophoretic peptide mapping with lysyl-endopeptidase indicated the non-identity of the multiple subunits of the 26S proteasome. From these structural and functional characteristics, the 26S proteasome, which is widely distributed in mammals, is suggested to be a new type of multi-molecular complex catalyzing the soluble energy- and ubiquitin-dependent proteolytic pathway.

The concentrations of free amino acid and protein-bound amino acid in gastric and colon cancers and also in normal tissues were measured and compared in order to investigate the differences in amino acid metabolism between these cancers. The examined materials were 20 specimens of gastric cancer, with 20 specimens of normal gastric full-thickness layers and 11 specimens of normal gastric mucosal layers, as well as 16 specimens of colon cancer, with 12 specimens of normal full-thickness layers and 16 specimens of normal colon mucosal layers. The amino acid pattern in gastric cancer was more similar to that in the gastric full-thickness layer than the gastric mucosal layer. On the other hand, the pattern in colon cancer was more similar to the colon mucosal layer than the full-thickness layer of normal colon tissue. Comparison between gastric and colon cancers showed that almost all of the protein-bound amino acid levels in gastric cancer were significantly higher than in colon cancer, while, conversely, all of the free amino acid levels except for aspartate, glutamate and glutamine were significantly lower in gastric cancer than in colon cancer. Both in the stomach and colon, the aspartate and glutamate levels in cancer tissues were significantly higher than in normal full-thickness or mucosal layers.

A 19 month old male infant with a mesenchymal hamartoma of the liver underwent an extended right hepatectomy. Serum bilirubin gradually rose until 3 months after the surgery, and obstructive jaundice and acholic stools were manifested at 6 months. Percutaneous transhepatic cholangio-drainage was performed. Cholangiography showed dilation of the intrahepatic bile duct of the residual lateral segment and complete obstruction of the extrahepatic bile duct. A second operation for reconstruction of the biliary tract was performed 10 months after the first surgery. No aspect of an extrahepatic biliary tract was found. Histological inspection of a surgical specimen of remnant tissue revealed only cicatricial connective tissue without any biliary structures. The clinical course has been uneventful for 18 months since the second surgery. The cause of bile duct atresia in this case is strongly suggested to be an ischemic change due to devascularization of the extrahepatic biliary tract following hepatic resection because of a tumor. To prevent this kind of complication, hepatico-enterostomy should be performed close to the cut surface of the liver.

A case is reported of extrahepatic obstruction of the portal vein with the unusual complication of obstructive jaundice that resulted from compression of the common bile duct caused by cavernous transformation of the varicose network. It is surgically impossible to repair choledochal stenosis resulting from enlarged varices in the vicinity of the common bile duct. In this case, internal drainage from the biliary duct to the duodenum using a silastic tube and a reservoir implanted in the subcutaneous space successfully eliminated the jaundice.

To learn the reasons for the high incidence of biliary carcinoma in patients with anomalous arrangement of the pancreaticobiliary duct (APBD) mutagenicity of the bile of APBD-modeled dogs that had received a dorsal pancreatico-cholecystostomy was assayed by the Ames Salmonella mutation test. The bile from two out of 18 APBD dogs was mutagenic for Salmonella typhimurium strain TA98 under the condition of metabolic activation by rat liver S9 fraction, while the bile from 17 normal dogs was not mutagenic. Furthermore, the bile from five APBD dogs i.p. administered 1-nitropyrene (1-NP), which is a typical environmental mutagen, was more mutagenic for strain TA98 than that from 1-NP-treated normal dogs. The bile from the APBD dogs had very high amylase activity, indicating that the bile contained pancreatic juice as a result of the pancreatico-cholecystostomy. When pancreatic juice from a normal dog was added to the bile from 1-NP-treated normal dogs, mutagenicity of the bile increased 1.6- to 2.0-fold. Furthermore, sulfatase increased the mutagenic activity of the bile in the presence of the pancreatic juice. HPLC revealed that the bile from a 1-NP-treated APBD dog contained mutagenic 1-nitro-6/8-hydroxypyrene and 1-nitro-3-hydroxypyrene, while bile from a 1-NP-treated normal dog did not contain these deconjugated products. The pancreatic juice from a normal dog had very high gamma-glutamyltransferase (GGT) and aminopeptidase activities and low sulfatase activity, but it had no beta-glucuronidase activity. In addition, the bacteria that easily infect the biliary duct of APBD dogs, Escherichia coli, Klebsiella, Enterobacter and Proteus, had high beta-glucuronidase activity. In particular, Klebsiella showed a very high sulfatase activity. These results suggest that pancreatic juice enzymes and bacteria infecting the biliary duct deconjugate the detoxified mutagens in the bile and induce mutagenicity of the bile from APBD dogs or APBD patients.

Two cases with an anomalous arrangement of the pancreaticobiliary ducts (APBD) are reported in order to investigate the mechanism of carcinogenesis of the extrahepatic biliary tract from the aspect of DNA ploidy analysis. Highly elevated biliary enzymes were found in both cases. Although inflammatory changes without any metaplasia or neoplasia were observed histologically, a cytofluorometric nuclear DNA ploidy analysis showed an aneuploid low ploidy pattern in a histogram of the gall bladder, an aneuploid high ploidy pattern in a histogram of the common bile duct in one patient, and a diploidy pattern in the gall bladder, an aneuploid low pattern in the common bile duct in another case. These results may show that APBD may play a role in the development in DNA ploidy abnormality with refluxed pancreatic juice which may induce repeated irritation and inflammation. Radical surgery including a complete excision of the extrahepatic biliary tract followed by reconstruction may achieve better results with regard to undesirable consequences with an abnormal nuclear DNA ploidy pattern and decrease the likelihood of developing carcinoma.

A 9-year-old boy with a painless reducible swelling in the right groin was admitted to our hospital. The diagnosis of a right external inguinal hernia was made, but no inguinal hernial sac was found at operation. On further examination, the diagnosis of femoral hernia was confirmed. Radical operation for the femoral hernia was performed using McVay's procedure, and there has been no recurrence since the operation. In the Japanese literature, 25 cases of femoral hernias have been reported in children aged 1 month-9 year (average age, 4 years). The sex ratio of M:F was 2:3. Eleven cases were on the right side, 7 were on the left, and 5 were bilateral. The rate of irreducible femoral hernia was 48% (12/25 cases). Femoral hernias in children are very rare and often misdiagnosed as inguinal hernias. Correct diagnosis was made in only three cases (12%). Careful examination and awareness of the possibility of femoral hernia in children are very important when dealing with groin swelling in children.

Three patients, i.e., two adults and one child, with unusual benign stricture of the extrahepatic biliary duct are reported. Case 1 is a 63-year-old male with obstructive jaundice caused by a sclerotic vascular ring formed by the right hepatic artery and its branch. Case 2 is an 18-year-old male with obstructive jaundice due to stricture of the common bile duct caused by compression of enlarged collateral varicose veins, so-called cavernous transformation following extrahepatic obstruction of the portal vein. Case 3 is a 19-month-old boy complicated with biliary obstruction resulting from ischemic changes of the biliary duct at 10 months after extended right hepatectomy for mesenchymal hamartoma of the liver. The definitive diagnosis was made on the basis of imaging evaluations and the surgical findings. They have been free of jaundice and have shown an uneventful clinical course after the surgical interventions.

We present a case of Peutz-Jegher's syndrome in an 18 year old female who was followed for gastrointestinal polyps for 13 years from the age of 5 years. The patient was treated four times with surgical or endoscopic polypectomy for gastrointestinal polyps. At the age of 14 years, a combined surgical and endoscopic approach for the management of Peutz-Jegher's syndrome was carried out. A large polyp of the ileum required enterotomy for its removal, and another smaller polyp of the upper jejunum was identified and removed by intra-operative total enteroscopy via the anus. Intra-operative enteroscopy allows one to identify polyps that would previously have been missed. A more complete polypectomy can be performed using this technique, allowing the patient with Peutz-Jegher's syndrome a longer interval between laparotomies and a reduction in symptoms attributed to polyps.

A 45 day old boy presented with progressive abdominal distension, tarry stools and anemia. A plain roentgenogram of the abdomen showed irregular, coarse calcifications in the left upper quadrant. The ultrasonography and computerized tomography of the abdomen revealed a large heterogeneous tumor with calcified parts in the left hemiabdomen. At operation, a 12 cm x 11 cm x 10 cm, multilobular, exogastric and endogastric tumor, including a portion of the anterior wall of the stomach, was extirpated. The pathological examination revealed a gastric teratoma with immature neural elements. Eight years after the total excision of the tumor there has not been any recurrence. This study includes a review of 88 cases (including the present case) of gastric teratoma reported in the English and Japanese literature.

A new classification of the anomalous arrangement of pancreaticobiliary ducts (APBD) has been proposed following amendments and modificiations of the previous one. Fifty-one cases of choledochal cyst complicated with APBD were extensively examined and analyzed for clear visualization of the APBD system to make a standard classification. APBD were classified broadly into three types: type I, type II, and type III with their subtypes. Type I of APBD was seen in 18 (35.3%), type II in 11 (21.6%) and type III in 22 (43.1%) cases, respectively. A radical operation, including complete excision of the dilated biliary duct followed by reconstruction of the biliary tract was the surgical treatment of choice, because the reflux of the pancreatic juice into the biliary tract is prevented. However, a dilated common channel or accessory pancreatic duct, according to the new Komi type Ib, IIb, and IIIc3 of APBD, could be the cause of relapsing pancreatitis leading to chronic pancreatitis, due to the formation of a protein plug or pancreatic calculus in the dilated duct, even after this radical operation. In two complicated cases of type IIIc3 of APBD in choledochal cyst, we performed a pylorus-preserving pancreatoduodenectomy as one of the operative methods of choice. Long-term follow-up, more than decades, is essential to evaluate the results of surgical procedure for choledochal cyst, especially in those complicated cases with type Ib, IIb, and IIIc3 of APBD according to the new Komi's classification.

An experimental model of anomalous arrangement of the pancreaticobiliary ducts (APBD) was produced by pancreaticocholecystostomy performed in 29 mongrel dogs. Our purpose was to study carcinogenesis of the extrahepatic biliary tract by DNA ploidy analysis with cytofluorometry. The amylase level in the bile was elevated in all 25 dogs tested except the controls. The phospholipase A2 level in the bile was elevated in all selected dogs except the controls. Common bile duct (CBD) dilatation was found in 23/29 (79%) of the dogs, and biliary stones were found in 3/29 (10%) dogs. Inflammatory changes were observed microscopically in all specimens except those from the controls. Intramural glandular structures were found in 17/25 (68%) of gallbladder (GB) specimens and 10/25 (40%) of CBD specimens; goblet cells were found in 7/25 (28%) of GB specimens and 2/25 (8%) of CBD specimens. In the controls neither glandular structures nor goblet cells were observed except for two GB specimens showing mild cholecystitis. Cytofluorometry showed 21% GB and 7% CBD diploidy, 69% GB and 65% CBD low ploidy, 10% GB and 28% CBD high ploidy patterns of histograms. These results show that, APBD may be central to high risk condition or play a key role to develop atypical biliary tract epithelium and DNA ploidy abnormality with or without biliary duct dilatation.

A 17 year old girl, with a papillary and cystic tumor of the pancreas, probably concealed within a previous post-traumatic pseudocyst of the pancreas is described. At 10 years of age, she had received a drainage procedure for a pancreatic pseudocyst, following a blunt abdominal trauma. The histological examination of the cyst wall did not show an epithelial lining. Seven years after that, she developed anemia and a computer tomography and ultrasonography of abdomen revealed a 10 cm x 9 cm x 8 cm, cystic, multilocular pancreatic mass with solid parts. On operation, a fist-sized, solid and multilocular cystic tumor, located in the body and tail of the pancreas, and infiltrating into the colonic serosa but with no metastasis, was found and completely excised. Histologic and electron microscopic examination revealed the characteristic features of a papillary and cystic tumor of the pancreas. This report suggests that cystic lesions of the pancreas should be carefully checked to decide the best surgical therapy.

This study was designed to investigate whether a combination of a glutamine antagonist (DON) and a diet deficient in glutamate and aspartate (AG) altered glutamine metabolism in tumor tissue, and inhibited tumor growth. In experiment-1, 21 male Donryu rats were fed with AG and implanted with Yoshida's Sarcoma. Of them, 7 rats were sacrificed on the 5th day (group AG), other 7 were sacrificed next day (group AG-1) and the remaining 7 were injected with DON on the 5th day and sacrificed next day (group AG+D). The tumor weight of group AG+D was significantly lower than of group AG, or of group AG-1. In experiment-2, of 23 rats, 9 were fed with control diet and 14 were fed with AG and implanted. 12 were sacrificed on the 5th day (group C, AG), and 11 were injected with DON on the 5th day and sacrificed next day (group C+D, AG+D). The reduced ratio of tumor weight in group C+D and group AG+D were 25% and 67%, respectively. These results show that the tumor growth could be inhibited by using metabolic antagonist of glutamine, and that it had synergistic effect in conjunction with the imbalanced diet.

The cytofluorometric nuclear DNA ploidy analysis was performed in gallbladder (GB) epithelium of eight patients of chronic cholecystitis to make a comparison between fresh and paraffin embedded tissue analysis, to make an ideal single cellular suspension, to evaluate the ploidy pattern affected by inflammation and to assess the histological correlations. Six (75%) fresh specimens and eight (100%) paraffin embedded specimens showed diploidy pattern of histogram. Two (25%) fresh specimens showed low ploidy pattern of histogram. There was a good correlation between histological findings and the ploidy patterns of 100% paraffin embedded specimens and 75% fresh specimens. The relative fluorescence intensity was found higher and more stable in paraffin embedded tissues than fresh tissues. Inflammation did not affect the ploidy patterns of paraffin embedded specimens whereas the ploidy patterns of fresh specimens were affected. It can be concluded that the DNA ploidy patterns and histological findings have a good correlation and inflammation does not affect the cytofluorometric DNA ploidy pattern when using the paraffin embedded specimens. Paraffin embedded specimens are superior to fresh specimens for making an ideal single cellular suspension to obtain stable, high, and accurate fluorescence intensity with less background fluorescence which makes the ploidy pattern behave like exact behavior of tissues.

Cytofluorometric nuclear DNA ploidy analysis was performed in two cases of primary hepatocellular carcinoma (HCC). This study was done to get the information about clinical behavior of tissues and to assess the prognosis. In both cases, almost all the liver function tests were abnormal, the tumors were thickly encapsulated, and the size of the tumors were below 5 cm in diameter. The DNA distribution pattern was categorized in two types, diploid and aneuploid with low and high ploidy, according to the mode and degree of dispersion of cells on the DNA histogram. Low ploidy pattern of histogram was found in one and diploidy pattern in another case. The serum alpha FP level was found high in case of diploidy pattern of histogram. The case of diploidy pattern of histogram died earlier than the case of aneuploid low ploidy pattern of histogram. The DNA ploidy patterns did not relate to the survival rates of patients. A preoperative cytofluorometric nuclear DNA ploidy analysis on biopsy specimens may be potentially useful to get the information about clinical behavior of tissues, to do the selective surgical procedures, and to assess the prognosis.

A nation wide survey for cystic fibrosis was carried out in Japan from 1982 to 1989. Cystic fibrosis was determined by extended criteria including high concentration of sweat chloride (more than 60 mEq/L), digestive, and respiratory manifestations, and family history. A total of 25 cases were confirmed as cystic fibrosis during the survey. Patients' characteristics were; age ranged from 0 day to 20 years old at diagnosis, sex ratio was 11:14 (M:F). All cases had respiratory, digestive symptoms and abnormally high sweat chloride excretion. Five cases from 3 families showed familiar involvement. These results show that cystic fibrosis is rare but really exists among Japanese. Further investigation including gene analysis should be carried out.

Cytofluorometric determination of nuclear DNA content in fresh gallbladder epithelium of 11 patients with biliary stones were performed to assess the histological correlations and to evaluate the ploidy pattern affected by inflammation or regenerative proliferating benign epithelium associated with non-neoplastic or neoplastic diseases. 27% of the patients were classified to have pure cholesterol stones, 54% were classified to have mixed stones and 19% were classified to have black stones. All of the specimens show suggestion of chronic cholecystitis without any metaplastic or neoplastic change. The bile culture shows free of bacteria in all of the cases. Cytofluorometric nuclear DNA ploidy analysis shows 64% diploidy and 36% low ploidy patterns of histograms. It can be concluded that the low ploidy pattern which could lead to false positive diagnosis of aneuploidy and may be associated with regenerative proliferating benign epithelium with severe inflammatory changes.

The activation mechanism of pancreatic enzymes refluxing into the biliary tract in the anomalous arrangement of pancreaticobiliary ducts (APBD) remains unclear. In order to elucidate this activation mechanism, an immunohistochemical examination of both bile ducts and gallbladders was carried out on 20 patients with APBD to determine whether or not enterokinase (EK) producing cells exist in the biliary tract, by employing an avidin-biotin-peroxidase complex (ABC) method using a monoclonal antibody, hek-1. Immunoreactive EK was found in the metaplastic epithelium of the bile duct in 2 patients and the gallbladder in one, suggesting that EK production at the metaplastic epithelium is involved in an activation mechanism of pancreatic enzymes refluxing into the biliary tract. The same study was performed on the gallbladders of 62 patients without APBD, which revealed immunoreactive EK in some parts of the metaplastic epithelium of the gallbladder in 2 patients. Thus, in cases of pancreatic juice refluxing into the biliary tract regardless of the presence of APBD, we can not refute the possibility that refluxed pancreatic enzymes may be at least partly activated by EK produced at the metaplastic epithelium.

Ingestion of foreign bodies is common in children. The majority of those which reach the stomach will pass uneventfully through the gastrointestinal tract. We report here an 9-month-old boy who swallowed an earring. Several roentgenologic examinations revealed a small, closed earring in the stomach. Endoscopic removal failed. At surgery the earring was seen grasping the pyloric ring and one end had eroded the gastric mucosa. These caused the earring to anchor firmly to the pylorus, as if it were attached to an ear lobe. The unusual way in which it was lodged in the stomach caused the impediment to its further progress, but fortunately, this was without serious consequences. Early endoscopic examination for retained foreign bodies in the stomach is recommended.

The metabolic response to the postoperative delivery of a sole glucose solution or a glucose-fructose-xylitol solution, and their effects upon carbohydrate metabolism, in patients with glucose tolerance impairment, were evaluated. Twenty four patients who showed an abnormal 75g-OGTT or were noninsulin-dependent diabetics were divided in two groups. The GFX-B group (n = 11) received a 10.5% solution, containing glucose, fructose and xylitol at 4:2:1 ratio, and the PHY-3 group (n = 13) received a 10% glucose solution, both, continuously infused for 3 days postoperatively. The infusion of PHY-3 tended to produce a higher increase in blood glucose than the infusion of GFX-B, that was significantly different on the 3POD (P less than 0.05). The increase in IRI and CPR was also significantly greater in PHY-3 group than in GFX-B group on the 3POD (P less than 0.05). We concluded that it was not only the slight lower glucose load of the GFX-B solution, but also, the different infused carbohydrate substrate which had influenced the magnitude of insulin secretion and the blood glucose level. Then, its use in glucose intolerant patients may be of some benefit to partially minimize the postoperative hyperglycemic response.

Cholestasis associated with total parenteral nutrition (TPN) has been reported to be an incidence of 15% to 50% in surgical neonates. This study was designed to prevent the liver dysfunction, especially intrahepatic cholestasis, caused by TPN for surgical neonates. In the last three years, 10 surgical neonates ranged from 1868g to 3500g of birth weight have been treated by cyclic TPN that infuses solutions of TPN and Non-TPN alternately every four hours, because cyclic TPN may reduce an overloading for the liver by the compulsive and continuous TPN. Consequently, no hyperbilirubinemia and abnormal values of the serum transaminases were revealed in these neonates, and good results of nutritional supports were obtained by cyclic TPN. These results suggest that cyclic TPN is useful and safe in nutritional support for neonates, and cyclic TPN is much better than continuous TPN in order to prevent the liver dysfunction.

Two siblings (a 2-year-old female and 11-month-old male) with similar onset of obstructive jaundice and clinical manifestations from early infancy are described. The jaundice fluctuated but never completely disappeared. Abnormal amounts of cholate, chenodeoxycholate and ursodeoxycholate were found in serum bile acid fractions. Pruritus, hyperbilirubinemia of predominantly the conjugated fraction and bilirubinuria were increased by repeated respiratory infections. Ultrasonography showed several highly echogenic shadows in the gallbladder in both cases, and gallstones were found at surgery. Operative cholangiography showed an anomalous arrangement of pancreaticobiliary ductal system in both cases. The pedigree showed five relatives (including the father) on the paternal side had liver disease, and an autosomal recessive inheritance was suggested. The association of familial intrahepatic cholestasis with a large amount of serum bile acids (which seem to be due to abnormal bile acid metabolism), cholelithiasis and anomalous arrangement of the pancreaticobiliary ductal system is proposed as a new hepatobiliary syndrome.

A monoclonal antibody, hek-1, was raised against enterokinase or enteropeptidase that had previously been partially purified from human duodenal fluid. Hek-1 showed staining of two glycoprotein bands of relative molecular weights of 260,000 and 240,000 on immunoblot analysis of partially purified enterokinase and of ammonium sulfate fraction of duodenal fluid. An enzyme immunoassay for human enterokinase was developed, making use of hek-1. Sensitivity to enterokinase was 20 times higher than that of the conventional assay where BAPA was used as a substrate. The immunohistochemical study with hek-1 showed staining of the brush border membrane and some goblet cells of the duodenum and upper jejunum but no staining of the colon epithelium.

In order to evaluate the efficacy of combined immunochemotherapy with mitomycin-C, tegafur, PSK and/or OK-432 as an adjunct for curatively resected gastric cancer, a prospective randomized controlled study using the envelope method was performed, in which 266 institutions from around Japan participated. The 3 year survival rates for all cases, and for ps(+).n(+) cases, were insignificantly higher in the immunochemotherapy groups receiving PSK and/or OK-432 than in the control group. However, because 28.2 per cent of the cases were excluded from the final statistical analyses, the results of this study may have questionable statistical credibility. Changes in the stimulation index (SI) suggest that the administration of PSK may result in an inhibition of the immunosuppressive activity of cancer patients. The high SI group showed a significantly higher 4 year survival rate than the low SI group.

Forty patients with choledochal cysts, 17 children and 23 adults, were treated by cyst excision in this institution (1972 to 1982). Histochemical and immunohistochemical studies were performed on tissue from the walls of these 40 cysts. The type and degree of histologic changes observed were found to correlate with patient age at time of cyst excision. Specimens from four patients less than two years of age at time of excision revealed epithelial desquamation, fibrosis, and minimal evidence of inflammation. Among specimens from 13 patients, 4 to 15 years of age, only two had an intact epithelial lining. Marked inflammation and intramural glandular structures were observed. Specimens from 23 patients, more than 15 years of age at the time of cyst excision, showed marked acute or chronic inflammation and destruction of the epithelial lining. In this age group, intramural glandular structures with goblet and argyrophil cells were regularly observed within the cyst walls, and gastrin and somatostatin-like immunoreactivity was detected in cells of these metaplastic glands. In association with these changes, in five of the 23 patients in the oldest age group, adenocarcinomas were observed in the cyst walls. These studies establish the increasing rate of epithelial metaplasia in the walls of choledochal cysts with advancing age, and suggest that these changes are a continuing process with malignancy one of the end results.

Addition of epinephrine to primary cultured adult rat hepatocytes stimulated their DNA synthesis dose-dependently, especially in presence of insulin and epidermal growth factor. This effect of epinephrine was strongly inhibited by an alpha 1-antagonist, prazosin, but not by a beta-antagonist, propranolol, and was also slightly inhibited by an alpha 2-antagonist, yohinbin. These results indicate that the stimulation of DNA synthesis of hepatocytes by epinephrine is mediated predominantly by an alpha 1-action. 12-o-Tetradecanoylphorbol-13-acetate (TPA) or Ca2+-ionophore A-23187 stimulated DNA synthesis of Swiss 3T3 cells, but did not induce DNA synthesis of hepatocytes either singly or in combination. The fact that pretreatment of hepatocytes with TPA caused down-regulation of the stimulatory effect of epinephrine on DNA synthesis of hepatocytes within 15 min suggested that the effect of epinephrine on hepatocytes is mediated by its alpha 1 receptor and that TPA activated protein kinase c in the hepatocytes. Addition of dibutyryl cGMP did not induce DNA synthesis of hepatocytes. Therefore, the alpha 1-action of epinephrine that induce stimulation of DNA synthesis of primary cultured adult rat hepatocytes was apparently not mediated by either activation of phospholipid-dependent protein kinase or Ca2+ mobilization. Possible alternative mechanism was discussed.

A nationwide survey of cases of choledochal cyst was conducted to obtain the data on cases of choledochal cyst in Japan between 1972 and 1982. A total of 645 cases in 73 institutions, including 52 cases from this hospital, were surveyed. Choledochal cyst was associated with an anomalous arrangement of the pancreaticobiliary ductal system in 90.2% of the cases which were examined extensively. The serum amylase level was elevated in 26.6% of the cases. Calculus disease of the biliary duct was recognized in 28.5% of the cases. Biliary carcinoma developed in 54 (8.4%) of 645 cases. The incidences of biliary carcinoma were 0.3% (1/306) in cases in children and 15.6% (53/339) in adult cases; carcinoma was found in the wall of the choledochus in 30, and in the gallbladder in 16 of these 54 cases. The nationwide survey showed that an anomalous arrangement of the pancreaticobiliary ductal system was closely associated with choledochal cyst and that cases of biliary carcinoma developed with the advance of age. Therefore, early resection of the cyst wall and reconstruction of the biliary duct is proposed as suitable surgical treatment for stopping reflux of pancreatic juice into the biliary duct.