Neutrophil recruitment in Clostridium difficile toxin A enteritis in the rabbit.

Neutrophil infiltration is a prominent feature of Clostridium difficile-associated enteritis and colitis. The aim of this study was to examine the importance of neutrophil recruitment and neutrophil-mediated tissue damage in C. difficile toxin A-induced enteritis. Competitive binding experiments using purified 3H-toxin A demonstrated the presence of a single class of medium affinity receptors on rabbit neutrophils (Kd 7 x 10(-8) M). Pertussis toxin and the nonhydrolyzable GTP analog GTPgamma S both inhibited 3H-toxin A binding (by 56 and 65%, respectively), indicating that the rabbit neutrophil toxin A receptor is G protein linked. Toxin A elicited a dose-dependent (25-200 micrograms/ml) stimulation of neutrophil migration in vitro, and this functional effect was also pertussis toxin sensitive (69% inhibition). Treatment of neutrophils with R15.7, a blocking monoclonal antibody to the leuocyte adhesion molecule CD18, inhibited toxin A-stimulated neutrophil migration by 85% in vitro. Pretreatment of rabbits with R15.7 also prevented neutrophil infiltration of toxin A-exposed ileal loops in vivo as determined by histologic examination and by ileal tissue myeloperoxidase levels. Furthermore, R15.7 effected a substantial inhibition of fluid secretion (by 65%), mannitol permeability (by 66%), and histologic damage in toxin A-exposed ileal loops. Anti-CD18 (R15.7) had no inhibitory effect on cholera toxin enterotoxicity. These data demonstrate that C. difficile toxin A is a proinflammatory toxin whose enterotoxic effects are substantially dependent upon neutrophil recruitment.

Study of the outcome of modified shoelace repair for midline incisional hernia.

AIMS AND OBJECTIVES: To study the efficacy and short-term sequelae of modified shoelace repair for midline incisional hernias. MATERIALS AND METHODS: A prospective non-randomized study of 30 cases of midline incisional hernias was carried out at a public hospital from May 2009 to Oct 2012. Patients underwent modified shoelace repair which comprises reconstruction of linea alba along with the use of polypropylene mesh to reinforce the facial layer. A proforma was maintained for each patient documenting patient details, nature of previous surgery and complications if any, postoperative course, and complications postincisional hernia repair. RESULTS AND DISCUSSION: Thirty patients underwent this repair with no complications and no recurrence for minimum follow-up period of 12 months. Our technique is a simple extra-peritoneal procedure with no extensive tissue dissection and avoids the potential complications of bowel injury and adhesions with the mesh. The principle involved is that during straining, the recti shorten by tonic contraction and approximate toward the midline. An incisional hernia weakens the midline and causes the recti to move laterally with contraction as medial pull is lost. By reforming a strong new linea alba, there is restoration of medial pull on recti. This is further buttressed by the mesh. CONCLUSION: Modified shoelace repair is a simple and safe extra-peritoneal procedure and can be used for all midline incisional hernias.

Genetic transformation of Vitreoscilla sp.

Of all the methods customarily used to transform E. coli we found only electroporation to be effective for transformation of the Gram-negative bacterium Vitreoscilla, yielding 5.10(5) transformants/microgram of plasmid DNA. The conditions used were close to those described for E. coli E. coli plasmids are stably maintained in Vitreoscilla. This is the first report of exogenous DNA transfer in Vitreoscilla which opens the way for the application of recombinant-DNA techniques to study this unique group of organisms.

Effect of bupropion on dopamine and 5-hydroxytryptamine-mediated behaviour in mice.

When bupropion (12.5-50 mg kg-1) was administered 30 min before methamphetamine it significantly antagonized methamphetamine-induced stereotyped behaviour in mice, but when given 5 min after methamphetamine it significantly potentiated the behaviour. When it was administered to mice pretreated with 100 mg kg-1 pargyline, intense locomotor stimulation and stereotyped behaviour was observed whereas when clomipramine was administered similarly the animals showed locomotor stimulation, head twitches and abduction and extension of hind limbs. Unlike clomipramine, bupropion failed to potentiate the 5-hydroxytryptamine-mediated behaviour seen after 5-hydroxytryptophan, 100 mg kg-1, i.v. These observations are in agreement with reports that bupropion is more potent as an inhibitor of dopamine uptake than as an inhibitor of 5-hydroxytryptamine uptake in-vitro.

Spontaneous perforation of the common bile duct in pregnancy.

Spontaneous perforation of the bile duct is rare. Bile duct perforation occurring during pregnancy is even rarer. We report a 21-year-old primipara with sealed bile duct perforation at 28 weeks of amenorrhea.

Gastrostomy without laparotomy--a percutaneous endoscopic technique: review of 35 cases.

Percutaneous endoscopic gastrostomy (PEG) was performed in 33 patient with head injury, one with laryngeal cancer, and one with gastric volvulus. The gastrostomy tube was prepared from 20 F Foley catheter and a plastic micropipette tip. The complications encountered included peritubal leak in three patients (9%) and abdominal wall hematoma in one patient (3%). There was no procedure-related mortality. We recommend PEG for tube enteral feeding in patients who have lost the swallowing reflex.

Gangrenous sigmoid volvulus in a pregnant woman.

A rare case of gangrenous sigmoid volvulus in a pregnant woman causing intestinal obstruction is reported. The patient had intrauterine foetal death. Laparatomy for resection of sigmoid colon and hysterotomy for removal of dead foetus was carried out. Terminal iliac colostomy with closure of rectal stump was done in the first surgery. The patient underwent colorectal anastomosis 2 months after the first operation and recovered uneventfully.

Massive rectal bleeding due to jejunal and colonic tuberculosis.

A case of massive rectal bleeding due to colonic tuberculosis in advanced pregnancy with intrauterine foetal death is reported. Patient was treated with resection of the left colon and left transverse end colostomy with closure of the rectal stump. Hysterotomy for the removal of the dead foetus was performed. The patient improved in health with antitubercular treatment. The colorectal anastomosis was performed after 4 months. Massive rectal bleeding in intestinal tuberculosis, though rare should be kept in mind.

Metoclopramide and experimentally-induced gastric ulceration in guinea-pigs.

Anti-gastric activity of metoclopramide was studied in guinea pigs using three different models of gastric ulceration. The effect of metoclopramide on gastric acidity was also studied. It was observed that metoclopramide affords protection against all types of experimentally induced gastric ulceration, without affecting the gastric acid secretion. The protective effect, therefore, is probably due to its ability to promote gastric drainage and to prevent the pyloric reflux, thus preventing corrosive effects of bile and acid on the stomach mucosa.

Isolated tuberculous hepatic abscess in a non-immunocompromised patient.

A 38 years old female presented with pain in the epigastrium, jaundice and fever since one and half month. The computerised tomographic scan of the abdomen revealed a multiloculated abscess of the left lobe of liver. The pus drained from the liver abscess at laparotomy showed acid fast bacilli on microscopy. A detailed search failed to identify any other focus of tuberculous infection. The case has been reported for the rarity of isolated hepatic tuberculous abscess and its presentation with jaundice, a rare feature, and to highlight the importance of microscopic or culture diagnosis in a suspected case of pyaemic abscess.

Ketotifen inhibits Clostridium difficile toxin A-induced enteritis in rat ileum.

BACKGROUND: Clostridium difficile toxin A is the principal mediator of inflammatory enterocolitis in experimental animals. The purpose of this study was to explore the effect of ketotifen, an anti-inflammatory drug, on toxin A-induced enterotoxicity in rat ileum. METHODS: The effects of intragastric administration of ketotifen on secretion, mannitol permeability, histological damage, and mucosal levels of leukotriene B4, leukotriene C4, and platelet activating factor in toxin A-exposed rat ileal loops were measured in vivo. The effects of ketotifen on toxin A-mediated release of rat mast cell protease II (rat mucosa mast cell product) release were also measured in rat ileal explants in vitro. The effect of ketotifen on neutrophil migration in vitro was also evaluated. RESULTS: Ketotifen pretreatment inhibited toxin A-associated intestinal secretion by 42.5% and mannitol permeability by 56.3% and reduced epithelial cell inflammation and necrosis. These effects were associated with reduced levels of leukotriene B4 by 65.8%, leukotriene C4 by 88.8%, platelet activating factor by 77.8%, and inhibition of rat mast cell protease II by 58.4%. In addition, pretreatment of neutrophils with ketotifen inhibited neutrophil migration in vitro. CONCLUSIONS: The protective effect of ketotifen in this animal model was associated with significant inhibition of release of mast cells and neutrophil derived mediators, supporting their involvement in C. difficile enteritis.

Saccharomyces boulardii inhibits Clostridium difficile toxin A binding and enterotoxicity in rat ileum.

BACKGROUND: Saccharomyces boulardii is a nonpathogenic yeast used for the prevention and treatment of Clostridium difficile-associated diarrhea and colitis. However, the mechanism by which S. boulardii exerts its protective effects remains unclear. METHODS: The binding of [3H]toxin A to its brush border receptor preincubated with S. boulardii-cultured suspension or filtered conditioned medium was measured in vitro. The effect of toxin A on secretion, epithelial permeability, and morphology in rat ileal loops in vivo was also examined in rats pretreated with S. boulardii. RESULTS: S. boulardii reduced [3H]toxin A-receptor binding in a dose-dependent fashion. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of ileal brush border exposed to S. boulardii-conditioned medium revealed a diminution of all brush border proteins. Treatment of rats with S. boulardii suspension reduced fluid secretion and mannitol permeability caused by toxin A. CONCLUSIONS: S. boulardii may reduce some of the enterotoxic effects of toxin A by inhibiting toxin A-receptor binding. This effect appears to be manifested by a secreted product of the yeast, possibly a protease.