The effect of laminin-1 on enteric neural crest-derived cell migration in the Hirschsprung's disease mouse model.

BACKGROUND/AIM: Laminin-1 regulates neurite outgrowth in various neuronal cells. We have previously demonstrated that laminin-1 promotes enteric neural crest-derived cell (ENCC) migration by using Sox10-VENUS transgenic mice, in which ENCCs are labeled with a green fluorescent protein, Venus. Mice lacking the endothelin-B receptor gene, Ednrb -/- mice, are widely used as a model for Hirschsprung's disease (HD). The aim of this study was to investigate the effects of laminin-1on ENCC migration in Sox10-VENUS+/Ednrb -/- mice, a newly created HD mice model. METHODS: Fetal guts were dissected on embryonic day 12.5 (E12.5). Specimens were incubated either with, or without laminin-1 for 24 h and images were taken under a stereoscopic microscope. The length from the stomach to the wavefront of ENCC migration (L-E) and the total length of the gut (L-G) were measured. Changes in the ratio of L-E to L-G (L-E/L-G) after 24 h were calculated. RESULTS: On E12.5, the wavefront of ENCC migration in the HD gut samples was located in the midgut, whereas the wavefront of ENCC in Sox10-VENUS+/Ednrb +/+ (WT) samples had reached the hindgut. After 24 h, L-E/L-G had increased by 1.49%, from 34.97 to 36.46%, in HD gut and had increased by 1.07%, from 48.08 to 49.15%, in HD with laminin-1, suggesting there was no positive effect of laminin-1 administration on ENCC migration in HD. CONCLUSIONS: Our results suggest that laminin-1 does not have a positive effect on ENCC migration in HD mice on E12.5, in contrast to the phenomenon seen in normal mice gut specimens, where laminin-1 promotes ENCC migration during the same period. This suggests that there is an impairment in the interaction between ENCC and extracellular environmental factors, which are required for normal development of the enteric nervous system, resulting in an aganglionic colon in HD.

Intraoperative colonoscopy facilitates safe dissection of the rectal pouch in a case of male imperforate anus.

INTRODUCTION: In an imperforate anus, colostography often fails to identify recto-urethral fistula (RUF). Thus, surgeons must always assume an RUF is present, despite colostography findings, and dissect the distal rectal pouch (RP) with caution. We report the usefulness of intraoperative colonoscopy (IOC) for excluding RUF and, thus, facilitating safe dissection of the RP. METHODS: We used IOC in six cases of imperforate anus. All had right transverse colostomy initially after birth. Distal colostography excluded RUF in five cases and was inconclusive in one. Laparoscopy was used to free the RP carefully from the bladder neck in all cases. Near the prostate, a 4-mm fine, flexible colonoscope was inserted into the RP through the anterior rectal wall to observe the laparoscopic dissection of the RP, which was attached closely to the prostate/bulbar urethra intraluminally to prevent injury to the urethra. The mucosa of the distal end of the RP was mucosectomized or diathermied, and the colon was pulled-through. Mean age at surgery was 11 months. RESULTS: IOC excluded RUF under direct vision in all cases, which enabled the dissection of the RP to be monitored and to proceed smoothly. At follow-up (mean: 31 months), all cases were well. CONCLUSIONS: IOC can be used to exclude RUF and facilitate safe dissection of the RP in imperforate anus.

Appendiceal stump closure in children with complicated appendicitis: a prospective analysis of endoloops versus endostaples.

INTRODUCTION: Laparoscopic appendectomy (LA) can be performed safely and effectively using endoloops (EL) or endostaples (ES). We compared EL and ES for stump closure during LA for complicated appendicitis in children. METHODS: All LA for complicated appendicitis performed between July 2005 and August 2009 were assessed prospectively. EL were used in 37 procedures and ES in 31. Apart from choice of technique which was the personal preference of the attending surgeon, all patients were managed according to the same intraoperative and postoperative protocols. RESULTS: There was no significant difference between mean age at operation; gender ratio; mean preoperative and postoperative white blood cell; mean preoperative and postoperative white blood cell C-reactive protein; histopathology; mean operating time (EL: 71 minutes; ES: 64 minutes); mean hospitalization (EL: 5.3 days; ES: 5.1 days); febrile period (EL: 2.1 days; ES: 1.9 days); white blood cell normalization (EL: 2.6 days; ES: 2.4 days); and intravenous antibiotic usage (EL: 3.8 days; ES: 3.7 days). There were no intraoperative complications or ICU admissions in either group, but two EL cases required conversion to open surgery (P=NS). Incidences of intra-abdominal abscess (EL: n=1 or 2.7%; ES: n=1 or 3.2%), transient ileus (EL: n=2 or 5.4%; ES: n=2 or 6.4%), small bowel obstruction (EL: n=0; ES: n=0), and wound infection (EL: n=1 or 2.7%; ES: n=1 or 3.2 %) were not significantly different. Rate of rehospitalization for EL was 2.7% (n=1; colitis), and for ES, it was 3.2% (n=1; intra-abdominal abscess) (P=NS). Mean cost for EL was US$890, and for ES, it was US$1300. CONCLUSION: This is the first prospective study comparing EL and ES during LA for complicated appendicitis in children. ES is more expensive, but there was no significant difference in morbidity for this technique, particularly with regard to incidence of postoperative intra-abdominal abscess.

Modified soave pull-through for Hirschsprung's disease: intraoperative internal sphincterotomy.

BACKGROUND/PURPOSE: Anorectal achalasia (AA) may persist after pull-through (PT) for Hirschsprung's disease (HD), which may cause postoperative enterocolitis (POE) and constipation. The authors modified the Soave PT (modified Soave PT, MSPT) to eliminate AA, and present their results. METHODS: This was a 16-year retrospective review of 43 patients with histologically proven HD of the rectosigmoid or sigmoid colon treated by MSPT. The MSPT involves excision of the posterior rectal cuff and an intraoperative internal sphincterotomy, allowing the PT colon to fit nicely. RESULTS: Mean age at MSPT was 16.7 months (16 were < or =3 months old [37%]; 7 were neonates [16%]). Mean follow-up was 9.2 years. Six of 43 cases (14%) had preoperative enterocolitis; only 2 of 43 (5%) had single episodes of POE. At review, 37 of 43 were older than 4 years; 29 (78%) had normal bowel function (14 had experienced soiling after MSPT, which resolved after a mean of 6.4 years); and 8 (21%) had problematic bowel function: 3 had occasional soiling, 1 had soiling only before defecation, 3 (8%) had constipation requiring laxatives or enemas, and 1 had significant soiling. CONCLUSION: MSPT is safe and may contribute to a reduction in the incidence of POE and constipation.

Does probiotics administration decrease serum endotoxin levels in infants?

PURPOSE: The aim of this study was to examine whether administration of probiotics to infants can change the ratio of intestinal flora and thereby decrease serum endotoxin produced by potentially pathogenic microorganisms. METHODS: Nine infants including five with of biliary atresia, two with omphalocele, one each with Hirschsprung's disease and imperforate anus were studied. All patients were stable, and no antibiotics were given during this study. A probiotic mixture consisting of Streptococcus faecalis, Clostridium butyricum and Bacillus mesentericus was administered orally to each infant at 2 g/day for 2 weeks. Fecal aerobic and anaerobic bacterial cultures, serum endotoxin level, and other biochemical parameters were examined. RESULTS: In fecal cultures, anaerobic bacteria including Bifidobacterium increased significantly whereas Escherichia coli, Streptococcus, and Klebsiella tended to decrease. The ratio of anaerobic to aerobic bacteria increased five times as a result of administration of probiotics, and serum endotoxin levels decreased. CONCLUSIONS: Probiotics affect intestinal bacterial flora by increasing anaerobic bacteria and decreasing the population of potentially pathogenic microorganisms. A decrease in luminal endotoxin may result in less endotoxin translocation or bacterial translocation.

The effect of epidermal growth factor on bacterial translocation in newborn rabbits.

PURPOSE: Epidermal growth factor (EGF), which is present in breast milk, has both trophic and maturational effects on intestinal mucosa. The aim of this study is to determine the effect of EGF on spontaneous intestinal bacterial translocation (BT) in formula-fed newborn rabbits, who have a high incidence of BT compared with breast-fed newborn rabbits. METHODS: Sixty-one rabbit pups were divided into three groups: EGF(-), n=24, EGF(+), n=22, and breast-fed animals, n=15. Both the EGF(-) and EGF(+) groups were gavage fed a standard artificial formula three times daily. EGF was administered subcutaneously three times daily (1.5 microg/g body weight per day) in the EGF(+) group. The breast-fed group was fed by their mothers ad libitum. At 7 days of age, all rabbits were killed, and the mesenteric lymph nodes (MLN), liver, and spleen were cultured qualitatively for bacterial growth, while the cecum and ileum were quantitatively cultured. To determine the effect of EGF on mucus-producing cells, goblet cell numbers in the small intestine were quantified histologically. RESULTS: There was no BT to MLN, spleen, or liver in the breast-fed group. The incidence of BT to MLN and spleen was significantly lower in the EGF(+) compared with EGF(-) group; (EGF[+]: MLN, 45%; spleen, 32%; Liver, 27%; EGF[-]: MLN, 79%; Spleen 67%; Liver 29%; in EGF[+] MLN and Spleen P<.05 vEGF[-]). There was no significant difference in cecal and ileal bacterial colonization between the EGF(+) and EGF(-) groups. The number of goblet cells in the small intestine was significantly lower in the EGF(-) group compared with the EGF(+) group as follows: EGF(+), 14+/-3; EGF(-), 9+/-3; breast-fed, 11+/-5 goblet cells per 100 epithelial cell nuclei; P=.013. CONCLUSIONS: (1) EGF caused a significant decrease in spontaneous bacterial translocation in formula-fed newborn rabbits and was associated with an increase in the goblet cell number of the small intestine. (2) These changes occurred in spite of the fact that no changes in small bowel bacterial colonization were observed. (3) These results suggest, but do not prove, that EGF may provide protection for neonates from gut origin infection by improving the mucosal barrier function through increased goblet cell production, thus decreasing the incidence of spontaneous bacterial translocation in the newborn.

Changes, with age, in the phospholipid content of the intestinal mucus layer of the newborn rabbit.

BACKGROUND/PURPOSE: The high incidence of bacterial translocation in newborns is thought to be caused, in part, by the immaturity of the intestinal mucosal barrier. Recently, intestinal mucus phospholipids (PL) have been reported to be important factors in the function of this mucosal barrier. The aim of this study was to quantify changes, with age, in the intestinal mucus PL of the newborn rabbit. METHODS: Mucus was gently scraped from the small intestinal mucosal surface of rabbits of different ages (0, 7, 14, and 28 days old and adult; n = 6 for all groups). PL was extracted from the mucus and was separated by two-dimensional thin-layer chromatography. The isolated phospholipid spots were quantified for their phosphorus content. RESULTS: Total PL content of the mucus decreased significantly with age (day 0, 21+/-2; day 7, 16+/-4; day 14, 9+/-3; day 28, 2+/-1; adult, 1+/-1 micromol/g wet mucus; P = .0001). Phosphatidylcholine and phosphatidylethanolamine levels in the adult rabbits were significantly lower in comparison with the 0-, 7-, and 14-day-old pups (P < .05). In contrast, lysophosphatidylcholine and lysophosphatidylethanolamine were significantly higher in the 28-day-old and adult rabbits in comparison with the 0-, 7-, and 14-day-old pups (P < .05). Phosphatidylinositol + phosphatidylserine levels in 7-day-old rabbits was significantly higher compared with adult rabbits. There was no significant difference in the composition of sphingomyeline between groups. CONCLUSION: Significant changes in the content and composition of the intestinal mucus phospholipids were observed during the first month of life in rabbits.

A comparison of the efficacy of pyloromyotomy and pyloroplasty in patients with gastroesophageal reflux and delayed gastric emptying.

PURPOSE: Delayed gastric emptying (DGE) in children with gastroesophageal reflux (GER) is often treated with a gastric emptying procedure. Although pyloroplasty is the most common gastric emptying procedure performed, pyloromyotomy is easier to perform and is associated with less morbidity. The aim of this study was to compare the efficacy of pyloromyotomy and pyloroplasty in children with DGE and GER undergoing a fundoplication. MATERIALS AND METHODS: We reviewed the charts of 54 patients with DGE who underwent pyloromyotomy (n = 29), or pyloroplasty (n = 25) along with a fundoplication. A technetium 99-labeled sulfur colloid liquid-phase gastric emptying study (GES) was performed in the pre- and early postoperative period (within 6 months after operation). Normal stomach emptying was defined as greater than 40% at 1 hour. Comparisons were made with regard to postoperative complication rate, incidence of redo fundoplication, length of postoperative hospital stay, and pre- and postoperative GES. RESULTS: The pyloroplasty and pyloromyotomy group were comparable in terms of age, sex, operative indications, and neurological status. There was no significant difference in the GES between the two groups preoperatively. There was a trend toward a decreased incidence of early postoperative complications including gas bloat, wound infection, pneumonia, dysphagia, bowel obstruction and dumping syndrome in the pyloromyotomy (8, 28%) when compared with the pyloroplasty group (12, 48%, P = .10). The mean postoperative hospital stay was 10.6 +/- 1.4 days for the pyloroplasty group and 7.6 +/- 1.0 days for the pyloromyotomy group (P + .08). The incidence of a redo fundoplication was 8% in the pyloroplasty and 7% in the pyloromyotomy group. Postoperative gastric emptying increased significantly in both groups (pyloroplasty group, from 18.1 +/- 3.1 to 49.5 +/- 7.9%, P = .0005; pyloromyotomy group, from 19.3 +/- 2.1 to 41.2 +/- 3.7%, P = .0001). There was no significant difference in the postoperative GES between the two groups (P = .289). CONCLUSION: Both pyloroplasty and pyloromyotomy performed in conjunction with a fundoplication resulted in a significant increase in early postoperative gastric emptying. There was no advantage of pyloroplasty over pyloromyotomy during this follow-up period. These data suggest that pyloromyotomy is an effective gastric emptying procedure in children with GER and DGE.

Intestinal permeability to small- and large-molecular-weight substances in the newborn rabbit.

BACKGROUND/PURPOSE: The authors have previously reported the occurrence of spontaneous bacterial translocation (BT) and its resolution with age in the newborn rabbit. They have also reported a close correlation between small bowel bacterial colonization (BC-SB) and BT at 1 week of age, suggesting that the presence of luminal bacteria and their production of endotoxins may increase the intestinal permeability. The aim of this study was to evaluate intestinal permeability to small and large molecules in the newborn rabbit and to correlate it with BT. MATERIALS AND METHODS: New Zealand White rabbits (n = 96) 1, 7, 14, 21, and over 120 days (adult) of age were given either C14-labeled ethylene diamine tetraacetic acid (EDTA) (MW 290) or C14-Dextran (MW 70,000) via an orogastric tube at 1 mCi per 100 g of body weight. Five hours later, blood, urine, liver, and intestine were collected, and scintillation counting was performed after solubilization. In a separate series of rabbits (n = 136), the incidence of BT, BC-SB, and small intestinal surface area (SA) were measured. RESULTS: Total permeability to Dextran decreased with age and was significantly reduced at 14 days of age. In contrast, total permeability to EDTA increased and was maximal in 7- to 14-day-old rabbits and began to decrease at 21 days of age. The incidence of BC-SB rapidly increased at 7 days of age and reached 100% at 14 days of age. The incidence of BT peaked at 7 days of life (30%) and then decreased with age. SA increased rapidly in the first 3 weeks and SA growth rate of 21-day-old rabbits was almost 1,400% compared with 1-day-old rabbits. CONCLUSIONS: This study has shown an age-related reduction of intestinal permeability to large (Dextran) and small (EDTA) molecular weight particles. However, intestinal permeability to EDTA had a different pattern than Dextran, suggesting that there may be different mechanisms of intestinal permeability to different size molecules. Intestinal permeability to EDTA closely correlated with bacterial colonization and bacterial translocation, suggesting that changes in the intestinal bacterial environment may affect the intestinal permeability, possibly by activating the immune system secondary to increases in endotoxins and bacteria.

The association of gut-associated lymphoid tissue and bacterial translocation in the newborn rabbit.

The authors have previously demonstrated spontaneous bacterial translocation (BT) in newborn rabbits and its resolution with aging. It is hypothesized that this spontaneous BT was associated with an immature gut-associated lymphoid tissue (GALT). The aim of the present study was to characterize the cellular populations of the GALT in rabbits at various ages and to correlate this with the frequency of BT. Small bowel (SB) sections and mesenteric lymph nodes (MLN) were harvested and cultured (aerobically) from New Zealand White rabbits at 0, 6, 14, 28, and more than 90 days of age for determination of bacterial colonization (BC) and BT. Portions of ileum (n = 6 for each age) were simultaneously harvested for immunoperoxidase staining. Total T cells (CDS+), expressed as the number of positive cells/1000 nuclei and activated T cells (CD25+), expressed as the number of positive cells/1000 nuclei and as the ratio of CD25+/CD5+ cells, were analyzed for each tissue. Positive cells were counted in 30 villi by light microscopy. The incidence of BT rose as BC increased in the small bowel and peaked at 6 days of age; BT then decreased with age. CD5+ cells in the small bowel villi at 0 days of age were few (2.5 positive cells/1000 nuclei) and the number significantly increased with age (6 days, 6.5; 14 days, 19.0; 28 days, 31.6; adult, 136.6 positive cells/1,000 nuclei). The distribution of T cells started in the crypts, and with advancing age, cells were found all the way to the top of the villi. The number of CD25+ cells in the villi increased with age. The CD25+/CD5+ ratio in the small bowel villi peaked at 6 days of age. These results demonstrate an inverse relationship between the number of CD5+ cells in the intestinal villi and the incidence of bacterial translocation. The elevation of activated T cells (CD25+) at 6 days of age may be the result of an immunologic activation during the time of peak bacterial translocation. These data suggest that maturity of the GALT leads to a loss of spontaneous bacterial translocation in the newborn period. Modalities that supplement the GALT may help reduce bacterial translocation.

Lingual thyroglossal duct cyst: a unique surgical approach.

Thyroglossal duct cyst (TGDC) is one of the more common causes of a pediatric neck mass. Lingual TGDC, which is located at the base of the tongue, is an unusual variant. Because of the oral pharyngeal location, lingual TGDC may cause dysphagia and respiratory distress. Previous investigators have advocated the use of a formal Sistrunk procedure for lingual TGDC. Herein the authors describe three children with a lingual TGDC in whom marsupialization of the cyst was performed, without excision. The follow-up period ranges from 2 to 5 years, and there has been no recurrence. Because of the low morbidity and high success rate associated with this approach, the authors recommend it for the treatment of lingual TGDC.

Acute hypoxia does not increase bacterial translocation in newborn rabbits.

PURPOSE: We have previously demonstrated that spontaneous bacterial translocation (BT) occurs in newborn rabbits and correlates strongly with small bowel colonization (BC). Birth stress, specifically hypoxia, is believed to increase this pathologic process and thus lead to sepsis. This study investigated the relationship between BT and acute hypoxia in newborn rabbits. METHODS: Four hundred seventeen rabbit pups (aged 0, 2 to 4, 6, and 28 days) were divided into four groups according to the type of hypoxic stress: 9% O(2) for 1 hour, 9% O(2) + 12% CO(2) for 1 or 4 hours, and 21% O(2) (control animals). The animals were killed 1.5 or 20 hours after the stress. Sterile specimens of mesenteric lymph nodes (MIN), spleen, liver, small bowel, and large bowel were incubated aerobically at 37 degrees C for 24 hours in thioglycolate broth, and subsequently plated on both MacConkey and Colistin Naladixic Acid media. After 24 hours, the growth on both plates was recorded. X(2) analysis was used, and P values of less than .05 were considered significant. RESULTS: BC of the small bowel and BT to the MLN were low in the first 4 days of life in the hypoxic groups (range, 0% to 21% BC, 0% to 6% BT) and the control group (range, 4% to 30% BC, 3% to 12% BT). After an increase in BC at 6 days of age, the rate of BT increased to 25% to 29% in control animals. The rate of BT in the hypoxic groups (25%) did not differ significantly from that of the controls (P > .05). Additionally, killing at 20 hours (v 1.5 hours) was not associated with an increase in the incidence of BT. None of the stress groups had a significant increase in BT compared with the controls. Importantly, although 4 hours of 9% O(2) + 12% CO(2) resulted in a 30% mortality rate, the incidence of BT was no higher than that of the control animals (13% v 29%; P > .05). CONCLUSION: Severe hypoxic stress in newborn rabbits does not increase the incidence of BT. Because the incidence of BT correlates with that of BC, and because BC is the same in the control and hypoxic animals, the sepsis observed in hypoxic newborns probably is not related to an increased incidence of BT.

Food restriction inhibits an autoimmune disease resembling systemic lupus erythematosus in (NZB x NZW) F1 mice.

To investigate the beneficial effects of food restriction on systemic lupus erythematosus in NZB x NZW F1 mice, we separated the mice into three groups. One was fed a diet in which total food intake was reduced to 60% of normal from age 2 mo onward, while the animals were still healthy (group 2R). A second group was selected at age 7 mo based on a positive lupus nephritis (proteinuria) and fed the 40% restricted diet thereafter (group 7R); a third group was allowed to consume food ad libitum (control). All control mice died of renal disease by age 14 mo, whereas all mice in group 2R and 80% of those in group 7R were living at that age. Measurements of anti-double stranded DNA antibody concentrations in sera and in supernatants of in vitro spleen cell cultures revealed that the production of the immunoglobulin G, but not immunoglobulin M, class of antibodies was markedly and significantly reduced in food-restricted mice. Age-associated changes in lymphocyte subsets seen in control mice, i.e., increases in B:T and CD4:CD8 T cell ratios, decreases in NTA260+ T cell subsets, and increases in aberrant activated NTA204+CD4+ T cells and cycling cells, were all significantly lessened in underfed mice. Food restriction did not suppress the secondary acquired antibody responses to a foreign antigen. Thus, the beneficial effects of food restriction in these mice may be related to the lessening of the age-related onset of T cell subset abnormalities, including activation of autoreactive T cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Determinant of bacterial translocation in the newborn: small bowel versus large bowel colonization.

This study investigated the relationship between bacterial translocation (BT) and small and large bowel bacterial colonization in the neonatal rabbit. The authors have previously shown that spontaneous BT occurs in rabbits during the first week of life, but is less frequent before the third day of life. The authors have also shown that bacterial colonization of the small bowel, is delayed until 3 to 5 days of age, and is preceded by colonization of the large bowel. Therefore, the authors hypothesizes that BT is dependent on small bowel, not large bowel, colonization. New Zealand White rabbit pups (n = 255) were divided into four groups, formula-fed (FF), formula-fed plus antibiotic (FFAB), breast-fed (BF) and unfed controls (UC). Animals from each group were killed on day 1 to 4 of life. Small bowel (SB) and large bowel (LB) specimens, mesenteric lymph nodes (MLN), spleen (SPL), and liver (LIV) were obtained from each rabbit, incubated for 24 hours in thioglycolate broth, and plated on both MacConkey and Colistin Naladixic Acid media in an aerobic environment. After 24 hours, growth on either plate was recorded. Bacterial colonization was significantly greater in FF versus UC, BF, and FFAB rabbit groups. In addition, large bowel colonization was significantly increased compared with small bowel colonization in all feeding groups (P < .05). Bacterial translocation to MLN, SPL, and LIV ranged from 10% to 27% in all groups. Gram-negative bacteria predominated over gram-positive bacteria in translocation to the various tissues sampled.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ursodeoxycholic acid treatment on essential fatty acid deficiency in patients with biliary atresia.

To assess whether ursodeoxycholic acid (UDCA) treatment has any beneficial effect on essential fatty acid (EFA) deficiency in patients who have had a Kasai operation for extrahepatic atresia (EBA), responses of serum fatty acids to UDCA administration (15 mg/kg/d) were investigated in eight jaundice-free patients and in eight patients with jaundice (serum total bilirubin > or = 1.0 mg/dL). All patients were also given taurine supplementation (100 mg/kg/d). Serum fatty acid composition was determined before and 6 months after UDCA treatment. Serum total bile acid concentration and serum total bilirubin value, as a part of conventional liver function tests, were measured before and during UDCA therapy. Before UDCA treatment, the concentrations of linoleic acid and arachidonic acid were significantly lower (P > .05 for the former; P > .01 for the latter) in both the jaundice and jaundice-free groups than in the controls. After 6 months of treatment, the linoleic acid concentration significantly increased (P > .05), to the normal range, in the jaundice-free group, but not in the jaundice group. The arachidonic acid concentration did not increase significantly in either group. The serum total bile acid concentration was lower in six of the eight jaundice-free patients and in four of the eight jaundice patients. The serum total bilirubin value decreased in six of the eight jaundice-free patients and in four of the eight jaundice patients; however, the degree of improvement was not statistically significant in either group. No side effects developed, and there were no changes in blood chemistry values unrelated to liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of neuromuscular junctions in the bowel affected by hypoganglionosis.

Using monoclonal antibody 171B5 against synaptic vesicle proteins, neuromuscular junctions and synapses in the muscle layers of the bowel affected by hypoganglionosis were labeled and their distribution analysed. In the hypoganglionic bowel, there were less synapses in the myenteric plexus and few neuromuscular junctions compared to normal bowels. The bowel dysmotility of the patient with hypoganglionosis appears to be due to inadequate innervation between ganglion cells and smooth muscle cells.

Hirschsprung's disease: diagnosis using monoclonal antibody 171B5.

A new reliable immunohistochemical method for diagnosing Hirschsprung's disease (HD) using our unique monoclonal antibody (MAb) 171B5 against synaptic vesicles is described. Fresh frozen sections of rectal tissues were used from 13 patients with HD aged 2 weeks to 13 months; 9 had rectosigmoid HD and 4 had total colonic aganglionosis (TCA). Comparable normal colonic and rectal specimens were also obtained from 13 age-matched controls. All specimens were labeled with MAb 171B5, to demonstrate neuronal innervation patterns of both mucosa and submucosa. In all control specimens, many synapses arranged in variciform plexuses were seen in the lamina propria, a moderate number in the muscularis mucosae, and dense clusters in the submucosal plexus. In all aganglionic specimens, only scanty numbers of synapses which were not organized in variciform plexuses were seen in the lamina propria, none in the muscularis mucosae, and a few in the submucosa. These findings suggest that MAb 171B5 immunohistochemistry on the lamina propria alone can differentiate between normal and aganglionic bowel and appears to be a reliable and useful method for detecting HD on suction rectal biopsy.