Effect of Endoscopic Sphincterotomy on Gallbladder Motility

In experimental animals, sphincterotomyfacilitates passage of solids from the gallbladder andinhibits gallstone formation apparently by improvementin gallbladder emptying. In humans, however, gallbladder emptying has not been studied followingendoscopic sphincterotomy (ES) in patients withgallstones. We therefore prospectively studied restingand cerulinstimulated gallbladder volumes by real timeultrasonography in 15 patients of choledocholithiasis withgallbladder in situ (eight with and seven withoutgallbladder calculi) before and after (after bile ductclearance) ES. ES significantly lowered restinggallbladder volume (21.2 ± 10.6 vs 11.1 ±5.0; P < 0.0001) and cerulin-stimulated residualgallbladder volume (10.8 ± 5.6 vs 4.4 ±2.1; P < 0.0001). ES also significantly increased thegallbladder ejection fraction (47.3 ± 12.1% vs 58.8 ± 11.1%; P < 0.0001). Therate constant for gallbladder emptying after cerulininfusion also increased significantly after ES(–0.022/min vs –0.031/ min; P < 0.0001).Significant improvement in gallbladder motility was observed in both groups ofpatients with and without gallbladder calculi. ESsignificantly improves gallbladder motility inhumans.     

Dendrosome-based delivery of siRNA against E6 and E7 oncogenes in cervical cancer

Although small interfering RNA (siRNA) treatment holds great promise for the treatment of cancers, the field has been held back by the availability of suitable delivery vehicles. For cervical cancer the E6 and E7 oncogenes are ideal siRNA targets for treatment. The purpose of the present study was to explore the potential of dendrosomes for the delivery of siRNA targeting E6 and E7 proteins of cervical cancer cells in vitro. Optimization of dendrimer generation and nitrogen-to-phosphate (N/P) ratio was carried out using dendrimer–fluorescein isothiocyanate oligo complexes. The optimized N/P ratios were used in formulating complexes between dendrimers and siRNA targeting green fluorescence protein (siGFP). Although formulation 4D100 (dendrimer-siRNA complex) displayed the highest GFP knockdown, it was also found to be highly toxic to cells. In the final formulation 4D100 was encapsulated into dendrosomes so as to mask these toxic effects. The optimized dendrosomal formulation (DF), DF3 was found to possess a siGFP-entrapment efficiency of 49.76% ± 1.62%, vesicle size of 154 ± 1.73 nm, and zeta potential of +3.21 ± 0.07 mV. The GFP knockdown efficiency of DF3 (dendrosome) was found to be almost identical to that of 4D100, but the former was completely nontoxic to the cells. DF3 containing siRNA against E6 and E7 was found to knock down the target genes considerably, as compared with the other formulations tested. Our results imply that dendrosomes hold potential for the delivery of siRNA and that a suitable targeting strategy could be useful for applications in vivo.From the Clinical EditorsiRNA treatment holds great promise for the treatment of cancers, but overall, the availability of suitable delivery vehicles remains a major issue. The purpose of this study was to explore the potential of dendrosomes for the delivery of siRNA targeting specific proteins in cervical cancer cells in vitro. The results suggest that dendrosomes hold potential for the delivery of siRNA and a suitable targeting strategy could be useful for applications in vivo.     

Role of hepatic lysosomes in the degradation of metallothionein.

The degradation of metallothionein (MT) by rat liver was examined. Degradation of MT by liver homogenate was greater than by cytosol. In addition, MT degradation by the homogenate at pH 5.5 was more than that at pH 7.2. Because lysosomal proteases function at acidic pH, these findings suggest the importance of lysosomes in MT degradation. The degradation by the lysosomal fraction was about 400-fold greater than that by the cytosol. Because cathepsins are the principal lysosomal proteases, we used cathepsin-specific inhibitors, such as leupeptin, E-64 and pepstatin, to determine the relative importance of different cathepsins in degrading MT. The study reveals that cathepsin B and/or L is (are) probably the most important enzyme(s) in degrading hepatic MT, because leupeptin, which blocks cathepsin B and L activity, inhibited the degradation of apo-MT by about 80%. Cathepsin D appears to be of least importance in MT degradation, because inhibition of this enzyme by pepstatin reduced degradation by only 20%. Studies on the degradation of apo-MT, ZnMT, and CdMT indicated that apo-MT is about 1500-fold more sensitive to degradation than ZnMT and CdMT. These data suggest that metals protect MT from degradation. This is further supported by a reconstitution experiment, which shows that with a progressive decrease of MT: metal ratio following titration of apo-MT by metals, there is a concomitant reduction in degradation. At a lysosomal pH of around 4.7, about 60% of Zn and 20% of Cd are displaced from MT, thereby making it susceptible to degradation. We propose, therefore, that lysosomes are probably important for MT degradation in vivo and that metal release is a prerequisite for degradation. With the release of metals, MT becomes susceptible to degradation, which is probably accomplished by the lysosomal cathepsins, in particular cathepsins B and L.     

Tropical calcific pancreatitis: strong association with SPINK1 trypsin inhibitor mutations

BACKGROUND & AIMS: Tropical calcific pancreatitis (TCP) is a chronic pancreatitis unique to developing countries in tropical regions. The cause of TCP is obscure. Whereas environmental factors, such as protein energy malnutrition and ingestion of cassava, have been implicated, a genetic predisposition to the disease also may be important. In the present study we report on mutations in the serine protease inhibitor, Kazal type 1 (SPINK1) gene in north Indian patients with TCP. METHODS: We studied 66 unrelated TCP patients (44 men, 49 with diabetes, and 6 with family history of TCP), 25 relatives, and 92 healthy control subjects. Samples were analyzed for SPINK1 variants (-53C>T, L14P, N34S, P55S, and 272T>C) and cationic trypsinogen (PRSS1) variants (A16V, K23R, N29I, and R122H) by melting curve analysis. RESULTS: Twenty-nine patients (44%) carried the N34S missense mutation, of whom 9 (14%) were homozygotes. In contrast, only 2 (2.2%) control subjects were N34S heterozygotes (prevalence ratio 20.2; 95% confidence interval 5.0-81.8; P < 0.0001 vs. TCP). The severity of pancreatitis did not differ between TCP patients with or without N34S, or among those heterozygous or homozygous for N34S. Among TCP patients with or without diabetes, the frequency of N34S carriers (43% vs. 47%) and N34S homozygotes (14% vs. 12%) was similar. CONCLUSIONS: TCP is highly associated with the SPINK1 N34S mutation. The high prevalence of N34S in TCP patients with and without diabetes suggests that these 2 subtypes have a similar genetic predisposition. The genetic predisposition to TCP resembles, at least in part, the idiopathic chronic pancreatitis found in industrialized countries.     

Predictors of septic shock following anastomotic leak after major gastrointestinal surgery: An audit from a tertiary care institute

Background:

Anastomotic leak is a serious complication after major gastrointestinal surgery and majority of deaths occur due to septic shock. Therefore, the early identification of risk factors of septic shock may help reduce the adverse outcomes.

Objective:

The aim of this audit was to determine the predictors of septic shock in patients with anastomotic leak after major gastrointestinal surgery.

Design:

Retrospective, audit.

Materials and Methods:

The patients admitted in the gastrosurgical intensive care unit ICU) of our institute between September 2009 and April 2012 with anastomotic leakage after surgery were identified. The ICU charts were retrieved from the database to identify the patients progressing to septic shock. A comparison of risk factors was made between the patients who developed septic shock (septic shock group) against the patients who did not (non-septic shock group).

Results:

The study sample comprised of 103 patients with anastomotic leak, of which 72 patients developed septic shock. The septic shock group had a higher APACHE II score, lower MAP, and higher HR at the time of ICU admission. They received greater transfusion of packed red blood cells during their ICU stay. Septic shock was more common after pancreaticojejunostomy and hepaticojejunostomy leaks.

Conclusion:

Presence of malignancy, chronic obstructive pulmonary disease (COPD), packed red blood cell transfusion, bacteremia, and hepaticojejunostomy or pancreaticojejunostomy leaks were independent predictors of mortality and length of ICU stay. To the best of our knowledge there are no available studies in the literature on the predictors of risk factors of septic shock in patients with anastomotic leakage.     

Duodenal bile examination in identifying potential non-responders to bile salt treatment and its comparison with gall bladder bile examination.

The results of bile salt treatment in patients with radiolucent stones and a functioning gall bladder have been poor. In 42 of these patients awaiting cholecystectomy we determined the value of duodenal bile examination in predicting gall stone composition, and thus identifying those less likely to respond to bile salt therapy. Based on chemical analysis and scanning electron microscopy, 28 of 42 (67%) gall stones retrieved at surgery were potentially insoluble. Microscopic examination of duodenal bile correctly identified 21 (75%) of them: it predicted all four (100%) pigment stones, three of six (50%) calcium carbonate containing cholesterol stones, and 14 of 18 (78%) cholesterol stones with pigment shells. It was nearly as reliable as microscopic examination of bile aspirated directly from the gall bladder during surgery (21 (75%) v 23 (82%); p = NS). Furthermore, the presence of cholesterol crystals in duodenal bile was a more sensitive indicator than chemical detection of supersaturation (34 of 38 (89%) v 25 of 35 (71%); p < 0.05) for prediction of cholesterol gall stones. Microscopic examination of duodenal bile, if used as a screening test, could help to exclude potential non-responders and thereby improve considerably the results of oral bile salt treatment for gall stone dissolution.     

Protective immunity to entamoeba histolytica infection in subjects with antiamoebic antibodies residing in a hyperendemic zone.

To investigate the relationship between amoebic gut infection and the presence of serum antiamoebic antibodies in a community where amoebiasis is highly endemic, a cross-sectional population survey was conducted covering 982 subjects in 2 adjacent villages in rural India. E. histolytica was detected in fecal smears in 18.1% (88/487) and antiamoebic antibodies by micro-ELISA in 28.7% (251/874) subjects. In 450 subjects in whom there was no clinical evidence of invasive amoebic disease or history of ingestion of antiamoebic drugs in the previous year and from whom both stool and matched serum samples were obtained, correlation of gut infection with seropositivity was attempted. Seropositivity in 80 cyst passers (23.8%) was significantly lower than in 370 non-cyst passers (35.1%; p less than 0.05). Conversely the rate of gut infection with E. histolytica was lower in seropositive subjects (19/149; 12.8%) as compared to seronegative subjects (61/301; 23%, p less than 0.05). In the population where E. histolytica infection was highly endemic, the rate of gut infection with the parasite was inversely related to seropositivity. Our data suggests that invasive E. histolytica infection (mostly asymptomatic) evokes good gut immunity in the host with clearing of the parasite from the colon and/or resistance to reinfection. High prevalence rate of amoebic antibodies indicates good "herd" immunity.     

Thoracoscopic lobectomy: is a training program feasible with low postoperative morbidity?

Objective

To evaluate the feasibility of training program in video assisted thoracic surgery (VATS) lobectomy comparing intraoperative and postoperative data of patients operated on by an established consultant and trainees.

Methods

Retrospective analysis of 100 consecutive patients who underwent VATS lobectomies between May 2008 and May 2012. 66 patients were operated on by an established consultant (Group A) and 34 by trainees (Group B).

Results

The groups were comparable for clinical characteristics and pathological staging. The mean operating time in Group A was 125 ± 30 min and in Group B was 133 ± 26 min (p = 0.18). The rate of conversion was similar in both groups: 9.1 % in Group A and 8.8 % in Group B (p = 0.6). The complication rate was comparable (p = 0.4): 36.3 % in Group A and 32.3 % in Group B. Median time to drain removal and median length of hospital stay was 3 and 5.5 days in Group A and 3 and 5 days in Group B, showing no statistical differences between the two groups (p = 0.3 and 0.5). There were no differences in term of long-term complications between the two groups.

Conclusion

Our study showed that a training program in VATS lobectomy is feasible, without increasing the operative time, conversion rate, postoperative complication, time to drain removal, and length of hospital stay.     

Effect of endoscopic sphincterotomy on gall bladder bile lithogenicity and motility

Background—Endoscopic sphincterotomyhas been shown to inhibit stone formation in the gall bladder ofexperimental animals.
Aims—To investigate the alterations in bilecomposition and gall bladder motility after endoscopic sphincterotomy.
Patients—A study was performed of gall bladderbile composition and gall bladder motility in patients with gallstonedisease ((n = 20; age 40-60 years, median age 55 years: seven men),with gall bladder calculi (n = 12) and with diseased gall bladder(chronic inflammation) without gall bladder calculi (n = 8)), who hadreceived endoscopic sphincterotomy for common bile duct stones. Age and sex matched disease controls comprised 20 patients with gallstone disease but without stones and an intact sphincter of Oddi (with gallbladder calculi (n = 10) and diseased gall bladder without gall bladdercalculi (n =10)).
Methods—Gall bladder motility was assessed byultrasound. Duodenal bile collected by nasoduodenal tube afterstimulation of gall bladder by intravenous ceruletid infusion wasanalysed for cholesterol, phospholipid, and bile acidconcentrations, cholesterol saturation index, and nucleation time.
Results—There was a significant reduction in mean(SEM) fasting volume (12.5 (1.7) ml v 26.4 (2.5) ml;p<0.001) and mean (SEM) residual volume (4.34 (0.9) ml v14.7 (0.98) ml; p<0.001), and increase in mean (SEM) ejection fraction(65.7 (4.2)% v 43.6 (5.52)%; p<0.001) and mean (SEM)rate constant of gall bladder emptying (−0.031/min v−0.020/min; p<0.01) in patients who had been subjected to endoscopicsphincterotomy. Median nucleation time was significantly longer (17 days v 6 days; p<0.006) in treated patients. There was areduction in total mean (SEM) lipid concentrations (6.73(0.32) g/dlv 7.72 (0.84) g/dl; p<0.05), cholesterol (5.6 (1.5) mmol/l v 10.3 (2.23) mmol/l; p<0.001) and CSI (0.72 (0.15) v 1.32(0.31); p<0.001). There was no significantchange in mean (SEM) phospholipid (25.6 (3.5) mmol/l v23.4 (6.28) mmol/l) and bile acid (93.7 (7.31) mmol/l v105.07 (16.6) mmol/l) concentrations.
Conclusions—After endoscopic sphincterotomy therewas enhanced contractility of the gall bladder, accompanied by aprolongation of nucleation time and reduction in cholesterol saturation index.

Keywords:gall bladder emptying; gall bladder contractility; nucleation time; cholesterol saturation index; gallstones; endoscopicpapillotomy