Prediction of contralateral inguinal hernias in children: a prospective study of 357 unilateral inguinal hernias.

PURPOSE: Previously, we established a pre-operative risk scoring system to predict contralateral inguinal hernia in children with unilateral inguinal hernias. The current study aimed to verify the usefulness of our pre-operative scoring system. METHODS: This was a prospective study of patients undergoing unilateral inguinal hernia repair from 2006 to 2009 at a single institution. Gender, age at initial operation, birth weight, initial operation side, and the pre-operative risk score were recorded. We analyzed the incidence of contralateral inguinal hernia, risk factors, and the usefulness of our pre-operative risk scoring system. The follow-up period was 36 months. We used forward multiple logistic regression analysis to predict contralateral hernia. RESULTS: Of the 372 patients who underwent unilateral hernia repair, 357 (96.0 %) were completely followed-up for 36 months, and 23 patients (6.4 %) developed a contralateral hernia. Left-sided hernia (OR = 5.5, 95 %, CI = 1.3-24.3, p = 0.023) was associated with an increased risk of contralateral hernia. The following covariates were not associated with contralateral hernia development: gender (p = 0.702), age (p = 0.215), and birth weight (p = 0.301). The pre-operative risk score (cut-off point = 4.5) of the patients with a contralateral hernia was significantly higher, compared with the patients without a contralateral hernia using the area under the receiver operating characteristic curve (p = 0.024). CONCLUSIONS: Using multivariate analysis, we confirmed usefulness of our pre-operative scoring system and initial side of the inguinal hernia, together, for the prediction of contralateral inguinal hernia in children.

Fine mapping and DNA fiber FISH analysis locates the tobamovirus resistance gene L3 of Capsicum chinense in a 400-kb region of R-like genes cluster embedded in highly repetitive sequences.

The tobamovirus resistance gene L(3) of Capsicum chinense was mapped using an intra-specific F2 population (2,016 individuals) of Capsicum annuum cultivars, into one of which had been introduced the C. chinense L(3) gene, and an inter-specific F2 population (3,391 individuals) between C. chinense and Capsicum frutescence. Analysis of a BAC library with an AFLP marker closely linked to L(3)-resistance revealed the presence of homologs of the tomato disease resistance gene I2. Partial or full-length coding sequences were cloned by degenerate PCR from 35 different pepper I2 homologs and 17 genetic markers were generated in the inter-specific combination. The L(3) gene was mapped between I2 homolog marker IH1-04 and BAC-end marker 189D23M, and located within a region encompassing two different BAC contigs consisting of four and one clones, respectively. DNA fiber FISH analysis revealed that these two contigs are separated from each other by about 30 kb. DNA fiber FISH results and Southern blotting of the BAC clones suggested that the L(3) locus-containing region is rich in highly repetitive sequences. Southern blot analysis indicated that the two BAC contigs contain more than ten copies of the I2 homologs. In contrast to the inter-specific F2 population, no recombinant progeny were identified to have a crossover point within two BAC contigs consisting of seven and two clones in the intra-specific F2 population. Moreover, distribution of the crossover points differed between the two populations, suggesting linkage disequilibrium in the region containing the L locus.

Semiquantative analysis of expression of mucosal addressin cell adhesion molecule-1 during small bowel graft rejection in rats.

AIM: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) mediates the homing of lymphocytes to gut-associated lymphoid tissues (GALT). We performed a semiquantative analysis of MAdCAM-1 expression during small bowel graft rejection. METHODS: Orthotopic small bowel transplantations (SBT) were performed from BN rats to LEW rats. Isografted animals served as controls. Animals were sacrificed on days 3, 4, 5, 6, and 7 after SBT. Cryostat sections were prepared from grafts, including Peyer's patches (PPs). Indirect immunoperoxidase staining was performed using mAbs against MAdCAM-1. The degree of vascular endothelial staining on high endothelial venules (HEV) in the PPs was graded from 1 (low levels) to 5 (high levels), and in the vessels of the lamina propria from 1 (faint), 2 (low at the base of villi), 3 (low to the middle of villi), 4 (high to the middle of villi), to 5 (high to villus tip). RESULTS: MAdCAM-1 expression on HEVs in PPs was down-regulated during rejection. In contrast its expression on endothelial cells of vessels in the lamina propria was up-regulated during rejection. CONCLUSION: Alteration in MAdCAM-1 expression may be associated with the development of SB graft rejection. The vessels at the base of villi, which are associated with lymphocyte recruitment, may become sites of intense immune reactivity during the early phase of small bowel allograft rejection.

Apoptosis of crypt cells and lymphocytes in gut-associated lymphoid tissues during small intestinal graft rejection in rats.

INTRODUCTION: We investigated the extent of apoptosis in crypt cells and Peyer's patches (PPs) during small bowel allograft rejection in rats to examine whether the Fas/FasL pathway participates in apoptosis within grafts during rejection. MATERIALS AND METHODS: Orthotopic small bowel transplantation with portocaval drainage was performed from Brown Norway to Lewis (LEW) rats. Isografted (LEW --> LEW) and nontransplanted animals served as the controls. Animals were sacrificed on days 3, 5, on 7 after SBT (each n = 5). An in situ end-labeling (ISEL) technique was used to detect apoptotic cells. Indirect immunoperoxidase staining was also performed using monoclonal antibodies against rat Fas or Fas-L. RESULTS: The number of ISEL-positive enterocytes in the allografts increased significantly on days 3, 5, and 7. Similarly, in the PPs of the allografts, the number of ISEL-positive mononuclear cells increased significantly on days 3, 5, and 7. On day 7 the number of Fas- and FasL-positive enterocytes were increased significantly in the allografts compared with the nontransplanted controls. Similarly, in the PPs, Fas- and FasL-positive mononuclear cells also increased significantly on day 7 in the allograft. CONCLUSION: Although an increase, number of apoptotic enterocytes and lymphocytes were observed in the early phase, activation of Fas/FasL system occurred during the late phase of small bowel graft rejection. These findings suggest that both rejection-associated and sepsis-induced forms of apoptosis may be associated with small bowel graft rejection.

Physiological studies on vasoactive intestinal peptide in rat small bowel isografts.

BACKGROUND: Vasoactive intestinal peptide (VIP) is released by stimulation of nonadrenergic noncholinergic (NANC) inhibitory nerves. In order to evaluate the function of VIP in jejunal isografts, we examined the enteric nerve responses in isografted rat jejunum compared with normal jejunum. METHODS: Orthotopic entire small bowel transplantation (SBT) with portocaval drainage was performed from Lewis rats to Lewis rats. Grafted tissue specimens were obtained 130 days after SBT (n = 8). As controls, normal segments of the jejunum were obtained from nontransplanted Lewis rats (n = 20). A mechanograph was used to evaluate in vitro jejunal responses to electrical field stimulation (EFS) of the adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers and VIP. RESULTS: The isografted jejunum was more strongly innervated by excitatory nerves, especially NANC excitatory nerves, than the normal jejunum (P <.05). VIP mediated relaxation reactions of NANC inhibitory nerves in the normal but to a lesser extent in the isografted jejunum (P <.05). CONCLUSIONS: The increased NANC excitatory nerves and the decreased effects of VIP in mediating NANC inhibitory nerves may largely relate to the peristaltic abnormalities seen in the isografted LEW rat jejunum.

Expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) during small-bowel graft rejection in rats.

BACKGROUND: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a critical endothelial adhesion molecule for lymphocyte trafficking to gut-associated lymphoid tissues (GALT) under physiological conditions. It is expressed on special postcapillary venules, that is the high endothelial venules (HEV) in GALT. In this study, we investigated changes in MAdCAM-1 expression during small-bowel graft rejection. METHODS: Orthotopic small-bowel transplantation (SBT) with portocaval drainage was performed from brown Norway (BN) rats to Lewis (LEW) rats. Isografted (LEW --> LEW) and untransplanted animals served as controls. Animals were killed on days 3, 4, 5, 6 and 7 after SBT. Cryostat sections (6 microm thick) were prepared from normal small-bowel tissues and small-bowel grafts, including Peyer patches (PPs). Indirect immunoperoxidase staining was performed using monoclonal antibodies (mAbs) against rat MAdCAM-1 (OST12). RESULTS: In the PPs of controls, MAdCAM-1 antibodies specifically stained the endothelial cells of HEV, which were predominantly located in the interfollicular areas. In the allografts on day 4 after SBT, the MAdCAM-1 expression was weaker on the HEV in the PPs than in the controls. In the lamina propria of controls, a faint expression of MAdCAM-1 on vessels was observed. On day 4 after allogeneic SBT, the MAdCAM-1 was more strongly expressed on the endothelial cells of the vessels at the base of the villi. As rejection developed, the MAdCAM-1 expression on the vessels progressed toward the villus tip. CONCLUSIONS: The change of MAdCAM-1 expression may be involved in the development of small-bowel graft rejection. The vessels at the base of villi, which is associated with lymphocyte recruitment, may become a site of intense immune reactivity in the early phase of small-bowel allograft rejection.

Analysis of Host Species Specificity of Magnaporthe grisea Toward Foxtail Millet Using a Genetic Cross Between Isolates from Wheat and Foxtail Millet.

ABSTRACT Host species specificity of Magnaporthe grisea toward foxtail millet was analyzed using F(1) cultures derived from a cross between a Triticum isolate (pathogenic on wheat) and a Setaria isolate (pathogenic on foxtail millet). On foxtail millet cvs. Beni-awa and Oke-awa, avirulent and virulent cultures segregated in a 1:1 ratio, suggesting that a single locus is involved in the specificity. This locus was designated as Pfm1. On cv. Ki-awa, two loci were involved and one of them was Pfm1. The other locus was designated as Pfm2. Interestingly, Pfm1 was not involved in the pathogenic specificity on cv. Kariwano-zairai. These results suggest that there is no "master gene" that determines the pathogenic specificity on all foxtail millet cultivars and that the species specificity of M. grisea toward foxtail millet is governed by cultivar-dependent genetic mechanisms that are similar to gene-for-gene interactions controlling race-cultivar specificity.

Analysis of pancreaticobiliary ductal union based on intraoperative cholangiography in patients undergoing laparoscopic cholecystectomy.

BACKGROUND: In order to diagnose pancreaticobiliary maljunction (PBM), it is necessary to perform direct fluoroscopic examinations of the biliary tract system. For patients with benign biliary diseases, these examinations are performed only in selected cases, because they are generally invasive. We investigated whether intraoperative cholangiography was practicable in evaluating the presence of PBM in patients with benign biliary diseases who underwent laparoscopic cholecystectomy (LC). METHOD: Between March 1998 and February 2001, intraoperative cholangiography (IOCG), which is one of the direct fluoroscopic examinations, was attempted in all 100 patients who underwent LC. RESULTS: IOCG was completed successfully in 98 patients (98.0%). No complications associated with IOCG occurred. In 13 (13.3%) of 98 patients, IOCG showed reflux of contrast medium into the pancreatic duct through a common channel. 'A long common channel' was observed in 8 cases (8.2%), suggesting the presence of PBM. CONCLUSION: These results suggest that IOCG in LC may be a practicable method for detecting the presence of PBM.

Novel glucoamylase-type enzymes from Thermoactinomyces vulgaris and Methanococcus jannaschii whose genes are found in the flanking region of the alpha-amylase genes.

A region downstream of the gene for pullulan-hydrolyzing alpha-amylase, TVA II, of Thermoactinomyces vulgaris R-47 was sequenced, and an open reading frame encoding an enzyme homologous to glucoamylase was found. The nucleotide sequence of this enzyme, designated TGA, consists of 1,953 base pairs corresponding to a protein of 651 amino acid residues. The TGA gene was subcloned and expressed in Escherichia coli. Enzymatic analyses showed that, like other glucoamylases, TGA produced beta-D-glucose from its substrate. However, TGA hydrolyzed maltooligosaccharides such as maltotetraose and maltose more efficiently than starch, while fungal glucoamylases preferred starch to maltooligosaccharides. The primary structure of TGA resembled a putative glucoamylase from the hyperthermophilic archaeon Methanococcus jannaschii (MGA), while homologies between TGA and the fungal glucoamylases were low. The enzymatic properties of recombinant MGA produced in E. coli cells were similar to those of TGA. These findings indicate that TGA and MGA are novel glucoamy-lase-type enzymes with oligosaccaharide-metabolizing activity.

Laparoscopic extended cholecystectomy.

BACKGROUND: Inadvertent dissemination of latent gallbladder (GB) cancer has become a crucial issue with the widespread use of laparoscopic cholecystectomy as a standard operation for benign gallbladder diseases. METHODS: Laparoscopic extended cholecystectomy is a new technique we have developed for the resection of GB tumors with suspicion of malignancy. The procedure is safe and minimally invasive and prevents tumor dissemination. The GB is resected laparoscopically along with a 1-cm margin of the liver using LaparoSonic Coagulating Shears without any ligation or clipping. RESULTS: We applied this technique to a patient with a GB tumor in the fundus. The tumor was not exposed on the cut surface, and there were no postoperative complications. CONCLUSION: Laparoscopic extended cholecystectomy for GB tumors with suspicion of malignancy is a minimally invasive technique that may decrease the risk of tumor dissemination.

A novel operative technique on proximal gastrectomy reconstructed by interposition of a jejunal J pouch with preservation of the vagal nerve and lower esophageal sphincter.

BACKGROUND/AIMS: This article describes the surgical techniques and postoperative status for proximal gastrectomy reconstructed by interposition of a jejunal J pouch with preservation of the vagal nerve and lower esophageal sphincter. METHODOLOGY: We have performed a new technique for reducing postgastrectomy sequelae such as reflux esophagitis, early dumping syndrome, and microgastria in early gastric cancer located in the proximal third of the stomach. The technique consists of proximal gastrectomy with preservation of the hepatic, pyloric, celiac branch of the vagal nerve, and abdominal esophagus (lower esophageal sphincter), and reconstruction by interposition of a jejunal J pouch. To reserve pyloric function, pyloroplasty can be omitted by preservation of the pyloric branch from the vagal nerve. To restore loss of reservoir function, the reconstruction is performed with an interposed jejunal J pouch. Sacrifice of the mesenteric arcades is kept to a minimum to preserve the autonomic nerve and blood flow in the mesentery. RESULTS: All of the patients who underwent this operation were able to eat an adequate amount of food at 6 months after surgery and they were satisfied with their postoperative status. And that, we have not experienced postgastrectomy disorders such the dumping syndrome and reflux esophagitis. CONCLUSIONS: Therefore, this method is useful for preventing the postoperative disorders in patients with early gastric cancer located in the proximal third of the stomach.

Prognostic studies on gastric cancer with concomitant liver metastases.

BACKGROUND/AIMS: The aim of this study was to clarify prognostic factors after surgical treatments in gastric cancer patients having synchronous liver metastases. METHODOLOGY: Clinicopathological features and prognosis were retrospectively reviewed in 43 surgical patients with gastric cancer with concomitant liver metastases from 1984 to 1998. RESULTS: More than half of the patients (51.2%) had numerous liver metastases (H3). Rates of peritoneal metastases (P1, P2, P3: 34.9%), tumor serosal invasion (T3 or T4: 73.8%) and widespread lymph node metastases (N3, N4: 69.8%) were also high. Although the gastric resection was performed in 29 patients (67.5%), concomitant hepatectomy resection was performed in only 3 patients (7.0%). Univariate analysis revealed that the grade of liver metastases (H1, H2, H3) was only a statistically significant prognostic factor (P = 0.008). Concerning surgical treatments, the resection group had tendency to better survival than the nonresection group (P = 0.074). Eight cases survived more than 1 year. Seven of the 8 cases (87.5%) were patients with gastrectomy. All of the 5 two-year survivors were patients with gastrectomy. CONCLUSIONS: In patients with gastric cancer with concomitant liver metastases, the grade of liver metastases indicate prognosis. In addition, there is possibility of palliative gastrectomy increasing the survival rare to more than one year.

The role of nitric oxide (NO) in the human internal anal sphincter.

PURPOSE: Nitric oxide (NO) has recently been shown to be a neurotransmitter in nonadrenergic noncholinergic (NANC) inhibitory nerves in the human gut. To clarify the physiological significance of NO in the human internal anal sphincter (IAS), we investigated enteric nervous responses in normal IAS muscle strips above the dentate line, obtained from patients with rectal cancer. METHODS: Normal IAS muscle strips above the dentate line, obtained from ten patients who underwent rectal amputation for low rectal cancers were used. The subjects consisted of eight men and two women, aged from 46-72 years (mean age, 54.2 years). A mechanographic technique was used to evaluate in-vitro IAS muscle responses to electrical field stimulation (EFS) of adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, N(G)-nitro-L-arginine (L-NNA) and L-arginine. RESULTS: Excitatory nerves were mainly involved in the regulation of enteric nerve responses to EFS in the baseline condition of the study, and NANC inhibitory nerves acted on the normal IAS. L-NNA concentration-dependently inhibited the relaxation in response to EFS in the human IAS, and this inhibitory effect in the IAS was reversed by L-arginine. CONCLUSIONS: These findings suggest that NANC inhibitory nerves play important roles in regulating relaxation of the human IAS, and that NO plays an important role as a neurotransmitter in NANC inhibitory nerves of the human IAS.

Operative technique on nearly total gastrectomy reconstructed by interposition of a jejunal J pouch with preservation of vagal nerve, lower esophageal sphincter, and pyloric sphincter for early gastric cancer.

Nearly total gastrectomy preserving the vagal nerve, the lower esophageal sphincter (LES), and the pyloric sphincter was developed as a function-preserving surgical technique to improve postgastrectomy disorders. In this paper, application criteria and technique are outlined, and postoperative quality of life was clinically investigated. Ten subjects who underwent this surgical operation (group A: 7 male and 3 female subjects at age 48 to 68 years with a mean age of 58.3 years) were interviewed to inquire about reflux esophagitis, dumping syndrome, and microgastria. Group A was compared with 20 cases of conventional total gastrectomy with D2 lymphadenectomy, excision of the lower esophageal sphincter (LES), total vagotomy, and single jejunal interposition (group B: 16 male and 4 female subjects at age 48 to 72 years with a mean age of 63.9 years). Included were cases with early cancer (M or SM1 of N0) localizing at the middle third and lower stomach, which was not applicable to endoscopic excision of gastric mucosa or partial gastric excision in M cancer, 2 cm or farther from the margin of the cancer to the esophagogastric mucosa cephalad junction and 3.5 cm or farther from the margin of the cancer to the pyloric caudad sphincter; in SM1 cancer, 4 cm or farther from the oral-side margin of the cancer to esophagogastric mucosa junction and 5.5 cm or farther from the anal-side margin of the cancer to the pyloric sphincter. In excision with lymph nodes, hepatic and celiac branches bifurcating from anterior and posterior trunks of the vagal nerve were preserved. To preserve LES, the esophagus was severed at the His angle at right angle to the longitudinal axis of the esophagus. The antrum was severed at 1.5 cm from the pyloric sphincter, preserving the arteria supraduodenalis. An alternative gaster was created as a 15-cm jejunal pouch with a 5-cm jejunal conduit for orthodromic peristaltic movement, using an automatic suture instrument to complete side-to-side anastomosis of folded jejunum with 1- to 1.5-cm long upper end of the pouch not anastomosed. The abdominal esophagus was mechanically anastomosed with a jejunal J pouch, and anastomosis of the pyloric antrum with a jejunal conduit was manually completed by stratum anastomosis. In group A, food ingestion per time could be taken the same as that of a healthy person, with no reflux esophagitis and dumping syndrome being noticed. Reflux esophagitis developed more significantly in group B than in group A (p < 0.05). In food ingestion per time, group B was significantly delayed compared with group A (p < 0.05). The present results suggested that the surgical technique proposed is a function-preserving gastric surgery appropriate to prevent postgastrectomy disorder of subjects.

Antisense oligodeoxynucleotides to human inducible nitric oxide synthase selectively inhibit induced nitric oxide production by human vascular endothelial cells: an experimental study.

OBJECTIVE: To find out whether the antisense oligodeoxynucleotide designed against the complementary DNA to human inducible NO synthase (iNOS) would block the translation from iNOS mRNA to NO in human vascular endothelial cells. DESIGN: Prospective controlled study. SETTING: Research laboratory, Japan. MATERIAL: Cultured human vascular endothelial cells. INTERVENTIONS: Human vascular endothelial cells were cultured for 24 hours, and divided into two groups, one of which was exposed to antisense oligodeoxynucleotides and the other to sense oligodeoxynucleotides at 100, 200, 400 and 800 micromol/L, respectively, for 18 hours. They were then exposed to interferon-gamma (1000 units/ml) and recombinant human tumour necrosis factor alpha (500 units/ml) to stimulate NO production. Each experiment was repeated ten times. RESULTS: Clear bands expressing cDNA of iNOS mRNA were identified in agarose gels in all cultured cells in both groups. The mean nitrite concentration in the supernatants of cultured cells in the group with the addition of antisense oligodeoxynucleotides was significantly lower than that in the group with sense oligonucleotides added at 200, 400, and 800 micromol/L. CONCLUSION: These findings indicate that the antisense oligodeoxynucleotides inhibited the NO production dependently, by preventing only their translation but not the processing, before their transcription from DNA into iNOS mRNA.

Relationship between gastroduodenal interdigestive migrating motor complex and quality of life in patients with distal subtotal gastrectomy for early gastric cancer.

A total of 40 patients (28 males, 12 females; mean age, 56.6 years; range, 41-72 years), 1-1.5 years (mean, 1.4 years) after subtotal gastrectomy for early gastric cancer (Billroth I, D2 lymph node dissection, curability A) were divided into 2 groups according to the occurrence of interdigestive migrating motor complex (IMMC), phase III from the duodenum, and their postoperative quality of life was compared. Results were as follows: (i) patients in the IMMC, phase III positive group (28 patients) had evidently more appetite and ate more food, with less decrease in body weight compared with the IMMC, phase III negative group (12 patients); and (ii) patients in the IMMC, phase III positive group had clearly less symptoms, such as early dumping symptoms (systemic symptoms), symptoms of reflux esophagitis (e.g. heartburn, feeling of regurgitation, difficult swallowing), nausea, abdominal pain, diarrhea, abdominal distention, and borborygmus, compared with the negative group. These results showed more satisfactory quality of life in the IMMC, phase III positive group compared with the negative group.