Appl Environ Microbiol 2003,69(1):290–296 CrossRefPubMed Authors’

Appl Environ Microbiol 2003,69(1):290–296.CrossRefPubMed Authors’ contributions LB designed the study, participated in all experiments, performed the analysis of CGH data, interpreted the results and wrote the manuscript. GF120918 LY carried out the Caco-2 invasion assays, plasmid extraction and participated in the analysis of data, the interpretation of results and the writing of the manuscript. MF carried out the CGH assays, and participated in the analysis of CGH data and in the correction of the manuscript. AM performed the PFGE

and RAPD experiments and participated in the analysis of data. NRT participated in the design of the study, collaborated in the interpretation of data and in the writing of the manuscript. AI participated in the design of the study and in the supervision of the analysis of CGH data. SP, CB, GA and FS

participated in the design of the study, the supervision of assays, and the writing of the manuscript. DM, SK and GD participated in the design of the study, the interpretation of results and the writing of the manuscript. JAC designed the study, supervised LB, LY and AM, participated in the analysis of data and interpretation of results and wrote the manuscript. All authors have read and approved the final manuscript.”
“Background The dimorphic fungal pathogen, Histoplasma capsulatum, parasitizes phagocytic cells of the mammalian immune system and causes one of the most common respiratory fungal infections world wide selleck screening library [1–3]. The mycelia-produced Histoplasma Selleck Ibrutinib conidia are acquired by inhalation into the respiratory tract where exposure to mammalian body temperatures triggers their differentiation into pathogenic yeast cells [3, 4]. Histoplasma virulence requires this transition to the yeast phase and expression of the corresponding yeast-phase regulon [5–7]. This transcriptional profile includes genes encoding specific factors that promote

Histoplasma virulence [7–9]. While mammalian alveolar macrophages efficiently phagocytose Histoplasma cells, they are unable to kill the yeast [10–12]. Within the macrophage, Histoplasma modifies the intracellular compartment to promote its survival and replication. The ability to subvert immune defenses and to survive within phagocytes enables Histoplasma to cause disease in both immunocompromised and immunocompetent individuals. This high potential for infection is reflected in the fact that histoplasmosis is one of the most common pulmonary fungal infections among healthy individuals [13]. The mechanistic details that CH5183284 clinical trial underlie Histoplasma pathogenesis are still largely unknown owing to limited or inefficient genetic methodologies.

For extraction

of secreted proteins, the supernatant was

For extraction

of secreted proteins, the supernatant was passed through a 0.2 μm Zap-cup sterile filter (10443401 Whatman Schleicher&Schuell) and proteins were precipitated with trichloroacetic acid (TCA, 10% [wt/vol] final concentration) over night at 4°C. The pellet was resuspended in 20 ml PBS in a 50 ml centrifuge tube (Falcon, BD) and vigorously mixed on a Vortex mixer (Vortex Genie 2, Scientific Industries) for 60 s at full speed in order to recover cell surface attached proteins (detached fraction). BAY 63-2521 nmr bacteria were harvested by centrifugation at 8,000 × g this website 30 min at 4°C. Residual bacteria were removed by passing the supernatant through a 0.2 μm filter (Corning) and proteins were precipitated with 10% [wt/vol] TCA over night at 4°C. The TCA precipitates

of the supernatant and the detached fraction were pelleted by centrifugation for 45 min at selleck compound 10,000 × g at 4°C. The pellet was washed twice with ice-cold acetone and recovered by centrifugation for 30 min at 10,000 × g at 4°C. The final pellet was air dried, resuspended in × μl sample buffer corresponding to the volume of the pellet and heated at 95°C for 5 min. Expression, surface-attachment and secretion protein profiles of wild-type SseB or SseD and mutant variants, were analyzed by SDS-Page using Tris-Tricine gels (12%) according to the method of Schägger and von Jagow [30]. For Western blotting, the semi-dry blotting procedure described by Kyhse-Andersen [31] was performed with slight modifications. The proteins were transferred onto 0.2 μm nitrocellulose membranes (Schleicher & Schüll) in Towbin buffer according to standard protocols [32]. For detection of SseB and SseD on Western blots, purified polyclonal rabbit antisera were used [7]. Mouse anti DnaK (Biotrend, Cologne, Germany) antibody was used to control equal loading of bacterial lysates as well as release of cytosolic protein into the detached fraction and the culture supernatant due to bacterial cell lysis. As secondary antibodies, horseradish find more peroxidase-conjugated

goat anti-rabbit IgG and goat anti-mouse IgG (HRP, Jackson) were used. The blots were incubated for 1 min with Pierce® ECL Western Blotting Substrate (32209, ThermoScientific) and exposed to X-ray films (Hyperfilm, GE, Freiburg, Germany). Cell culture and infection procedure For infection experiments, the murine monocyte cell line RAW264.7 was cultured in DMEM (E15-843, PAA, Pasching, Austria) supplemented with 10% FCS (Sigma-Aldrich) and 2 mM Glutamax (Invitrogen) at 37°C in 5% CO2and 90% humidity. The cells were used for experiments up to passage number 25. Cells were seeded in 24 well plates (Greiner bio-one) one day before infection and allowed to duplicate. Bacteria were grown overnight at 37°C and stored at 4°C until use. Cultures were adjusted to OD600 = 0.

The remainder of the paper is arranged as follows The next

The remainder of the paper is arranged as follows. The next

section briefly explains the construction of MD simulation models and introduces the indentation process parameters for the simulation cases. Thereafter, the simulation results under dry indentation and wet indentation are compiled. They include the comparisons of load–displacement curves, calculated hardness and Young’s modulus values, the distributions of friction and normal forces along the indenter/work interface, and stress distribution within the work material. Finally, conclusions are drawn in the final section. Methods Three-dimensional (3D) MD simulation models are constructed to study the indentation processes on single-crystal copper by a half-cylinder diamond indenter. CP-690550 molecular weight For wet indentation, water molecules are added to fill the gap between the indenter and the work material in the system. For dry indentation, no water molecules are added. We employ LAMMPS, an open-source software developed by Sandia

National Laboratory [21], to carry out the simulation computation. Figure 1 shows the schematic of MD simulation models for RG7112 mw dry and wet indentations. The dimension of the copper work material is 247 × 216 × 70 Å3 (X, Y, and Z directions, respectively) for all simulation cases, and it consists of 306,000 copper atoms. The copper indentation surface is a (1 1 1) plane. The indenter has a radius of 50 Å, consisting of 46,000 carbon atoms. At the initial stage, the offset distance between the indenter and the work material is 5 Å. For wet indentation cases, the entire indenter is submerged in water, so both the

indenter and the work material are in contact with water. In this case, 90,324 water atoms are contained in the system, including 60,216 hydrogen atoms and 30,108 oxygen atoms. Meanwhile, two special layers are defined in the copper work material, namely a thermal layer and a fixed layer. The fixed layer is located at the bottom of the work material, and it acts as a base to avoid Mannose-binding protein-associated serine protease any movement of the work material. The thermal layer is located right above the fixed layer, and it acts as a heat sink to maintain the temperature of the simulation system. In addition, since the simulation size is extremely small, a periodic boundary find more condition is applied along the Z direction so that the simulation box is replicated throughout the space to form an infinite lattice. This can effectively mitigate a spurious size effect when investigating the behavior of an isolated system. Figure 1 Schematic of MD simulation models for (a) dry nano-indentation and (b) wet nano-indentation. In this study, we compare wet nano-indentation with dry nano-indentation by focusing on the tool/material interaction and process performances. Meanwhile, we consider the potential effect of indentation speed by including three levels of indentation speed. As a result, six simulation cases are created. Table 1 presents the detailed parameters of the six cases.

The name was reinstated by Holm (1957) and was represented by N

The name was reinstated by Holm (1957) and was represented by N. hirta, which was

concurrently treated as a synonym of N. derasa (Berk. & Broome) L. Holm. The most outstanding morphological characters of Nodulosphaeria were considered to be apex of ascomata often covered with setae, selleck chemicals ascospore with three or more transverse septa with a supramedian enlarged cell or elongated to a scolecospore, mostly with terminal appendages (Barr 1992a; Holm 1961; Shoemaker 1984b). The ascomata are usually immersed and the peridium comprises a few layers of brown, relatively thin-walled cells of textura angularis and textura prismatica LY2874455 ic50 similar to those of Phaeosphaeria. Thus, Nodulosphaeria is likely to be a member of Phaeosphaeriaceae. However, this needs to be confirmed by molecular analysis. The boundary between Nodulosphaeria and Ophiobolus is not clear-cut, and the circumscriptions of them usually depend on the viewpoint of different mycologists. For instance, Shoemaker (1976) has assigned some Nodulosphaeria

species such as N. erythrospora, N. fruticum, N. mathieui and N. megalosporus to Ophiobolus. Subsequently, more species were added to Nodulosphaeria (Barr 1992a; Shoemaker 1984b; Shoemaker and Babcock 1987). Currently, more than 60 names are included in Nodulosphaeria (http://​www.​mycobank.​org/​, 06/2010). Phylogenetic study None. Concluding remarks Selleck RAD001 All species included in Nodulosphaeria have an inflated ascospore cell as mentioned above. However, it is likely that this character would have evolved more than once as it is probably an adaption for ascospore ejection from the ascus (Shoemaker 1976). It occurs in Ophiobolus species and the ascomata of these species are quite dissimilar to Nodulosphaeria species and their exclusion from Nodulosphaeria seems warranted.

When considering whether a species belongs in Nodulosphaeria, one must also consider the ascomata and peridium structure until DNA sequences are available. Ohleria Fuckel, Fungi rhenani exsic.: no. 2173 (1868). (Melanommataceae) Generic description Habitat terrestrial, saprobic. Ascomata small to medium size, solitary, scattered, or in small groups, erumpent to nearly superficial, papillate, ostiolate. Astemizole Peridium thin, thicker at the apex, 1-layered. Hamathecium of dense, long trabeculate pseudoparaphyses. Asci 8-spored, bitunicate, fissitunicate, cylindrical, with a short pedicel. Ascospore brown to reddish brown, broadly to narrowly fusoid, 3-septate, easily separating into two parts at the primary septum. Anamorphs reported for genus: Monodictys (Samuels 1980). Literature: Barr 1990b; Clements and Shear 1931; Patel et al. 1997; Samuels 1980. Type species Ohleria modesta Fuckel, Fungi rhenani exsic. (1868) (Fig. 68) Fig. 68 Ohleria modesta (from g: f. rh. 2173, isotype). a Ascomata scattering on host surface. b Section of a partial peridium.

The calcium supplements contained 1 g or more, and could have bee

The calcium supplements contained 1 g or more, and could have been taken in the fasting state. As mentioned by the authors, this selleck compound may give rise to transient hypercalcemia for several hours, which—when

repeated every day over several years—might increase the risk of coronary heart disease. Indeed, no increased cardiovascular risks were observed with calcium from food which is absorbed more slowly. Even the administration of a calcium supplement in the form of bone powder does not increase the plasma calcium level above normal [11]. In the same way, calcium supplements increase slightly the risk of renal stones in some studies, whereas calcium from food decreases this risk [2]. It might be assumed, therefore, in the light of the studies of Bolland

et al. [4, 5], that supplements of only 500 mg of calcium taken after a meal are harmless, even when taken twice a day. The question remains if a supplement of 500 mg per day is enough. One could argue that a supplement of 500 mg of calcium does not meet the requirements, which were redefined recently by the Institute of Medicine in the USA (IOM) [1]. The report states that 1,000 mg of calcium is the estimated average requirement for women over 50 years, and 1,200 mg/day is the recommended daily allowance. But these figures are derived from studies in populations whose bone health was not optimal. These studies were not titrated against the blood level of 25-hydroxyvitamin Pritelivir molecular weight D. They were performed in populations that probably were—as we now know to be—vitamin D deficient. Vitamin D deficiency is prevalent worldwide [12] and Rebamipide it is reasonable to assume, therefore, that the recommendations of the IOM are unnecessarily high. If human beings were exposed to sunlight regularly, not only would they have higher 25-hydroxyvitamin D levels, they might also need less calcium for optimal bone health. It is, by the way, surprising, how low the recommendations of the IOM report are for vitamin D. They were considered by experts like R.P. Heaney and M. Holick as to ‘fail on three grounds: logic, science and guidance’ [13]. This

TH-302 datasheet allows us to suppose that calcium supplements of 500 mg are effective, so long as the vitamin D level is optimal. Indeed, high 25-hydroxyvitamin D levels seemed to compensate for the otherwise negative effects of a low calcium intake (<716 mg/day) on BMD [14]. In conclusion, if the reported increased risk of MI induced by calcium supplements of 1,000–1,200 mg were the result of a meta-analysis of studies with MI as primary outcomes, it still would not challenge the clinical practice free of cardiovascular dangers, which favours supplements of 500 mg to be taken after meals, combined with vitamin D when the nutritional intake of calcium does not sum up to 800 mg. References 1. Report on Dietary Reference Intakes (DRIs) for calcium and vitamin D by the Institute of Medicine (IOM) (2011) Dietary reference intakes for calcium and vitamin D.

0 and pH 5 5, which was also found in strain SA45 The same expre

0 and pH 5.5, which was also found in strain SA45. The same expression pattern has been found for the prophage-encoded Panton-Valentine leukocidin (PVL, luk-PV) of S. aureus [28]. Maximal expression of luk-PV in the late exponential growth phase was followed by a rapid decline post-exponentially. Our observation could partially be explained by the induction of the prophage carrying the toxin gene. The sea-phage copy numbers of S. aureus Mu50 at pH 6.0 remained constant during the first part of cultivation. In the late stationary growth phase, however,

the number had increased four times (average GDC-0994 cost increase of two biological replicates) compared to levels in early stationary growth phase. The phage copy numbers might have increased further if growth was allowed to continue. An acetic-acid induced intracellular drop in pH, leading to oxidative stress [29] would activate the SOS response system inducing the prophage [30]. Sumby and Waldor showed that upon prophage induction in S. aureus, the phage DNA was replicated, resulting in an increase in sea gene copy number, and that a second prophage-regulated sea promoter was also activated, resulting in increased sea expression [14]. Similar enhanced transcription

of phage-encoded virulence genes upon prophage induction has also been observed for PVL in S. aureus and the Shiga toxins in E. coli [28, 31]. Mitomycin C, a well-known prophage inducer, was used in this study. The more MC added, the more SEA was produced per CFU for all three strains PU-H71 in vitro tested, supporting

the association between prophage induction and SEA production. However, the expected boost in extracellular SEA levels accompanying the increased sea mRNA levels and sea gene copy levels observed at pH 5.5 was not found. This could be because of the pronounced phage production at pH 5.5 seen as a rapid increase in extracellular sea-phage copy number (Figure 3). The window for phage-encoded SEA-biosynthesis prior to phage-release could be too narrow in the bacteria at this pH level. The relative phage copy number generally increased over time at all pH levels investigated. At pH 5.5, the relative phage copy number was increasing dramatically over time, suggesting that substantial prophage induction had occurred. The sea gene copy number, however, was decreasing over acetylcholine time at pH 5.5. This could be due to cell lysis occurring upon prophage induction at this pH. At pH 5.0 and 4.5, a big increase in relative sea gene copy number was observed between the two last sampling points. This suggests that the prophage has been induced and the replicative form of the phage DNA is produced. However, at these low pH values, no great increase in SEA or phage copies were observed, suggesting protein synthesis was impaired. In addition, the reason why the sea expression of S. aureus Mu50 at pH 5.5 was not as high as at pH 6.

coli DH5α containing only the pSUP202 vector

coli DH5α containing only the pSUP202 vector PI3K Inhibitor Library (Figure 3B). Further, find more phospholipase A2 activity was examined in various subcellular fractions prepared from E. coli strain S299, including cytoplasm, cytoplasmic membrane, and outer membrane fractions. Most Plp activity was detected in Tween-20 soluble membrane fraction, indicating that Plp was

mainly localized in the cytoplasmic membrane of E. coli S299 (data not shown). No BODIPY-labeled free fatty acid (FFA) (at sn-1 position) was detected in the TLC analysis when an apolar solvent was used (data not shown), and BODIPY-labeled LPC was not further degraded by Plp in the reaction, indicating that Plp had no lysophospholipase or phospholipase B activity. Figure 3 Thin-layer chromatography (TLC) demonstrates

phospholipase A2 activity of Plp. BODIPY-labeled phosphatidylcholine (BPC) was incubated with various standard enzymes or sample preparations for 1 h at 37°C. Subsequently, the lipids were extracted check details and separated by TLC. (A) The cleavage patterns of BPC by standard proteins PLA2, PLC, and PLD were able to distinguish the different phospholipase activities. (B) Cleavage patterns of BPC by supernatants (lanes 2 and 3) and cell lysates (lanes 4 and 5) from E. coli DH5α containing cloned plp (lanes 3 and 5) or just the cloning vector pSUP202 (lanes 2 and 4). Lane 1 contains only BPC incubated in the presence of PBS buffer. BLPC, BODIPY-labeled lysophosphatidylcholine; PA, phosphatic alcohol; PBt, phosphaticbutanol; DAG, di-acylglycerol. Enzymatic characteristics of rPlp protein To examine the enzymatic characteristics of Plp, the entire coding sequence of plp was cloned and inserted into the expression vector

pQE60, which adds a His6 (His-6×) tag to the carboxyl end of Plp. The over-expressed recombinant Plp (rPlp) formed inclusion bodies in E. coli. To recover 4��8C active rPlp, purification of the inclusion bodies followed by solubilization under mild conditions and re-folding was performed as described in the Methods. Purity of refolded rPlp protein was confirmed by SDS-PAGE and silver staining (data not shown). The final concentration of purified rPlp protein was 8 μg/ml with a recovery of <10%. Subsequently, the enzymatic characteristics of refolded rPlp were examined under various chemical and physical conditions. The enzymatic activity of rPlp positively correlated to its concentration from 1 μg/ml to 8 μg/ml (Figure 4A); therefore, 4 μg/ml rPlp protein was routinely used in other activity assays. The enzymatic activity unit of refolded rPlp (1 unit = amount of protein that cleaves 1 μmole of BODIPY-PC per minute) was about 2,500-fold higher than standard PLA2 enzyme extracted from porcine pancreas, which indicated that Plp had a high activity against the BPC phospholipid substrate. Plp enzyme activity exhibited a broad temperature optimum from 37°C to 64°C (Figure 4B) with 75% activity retained at 27°C and 50% activity at 20°C.

These results were validated using an approximate likelihood rati

These results were validated using an approximate likelihood ratio test in PhyML [45]. The phylogenetic tree of OMPLA conflicts with that

of AtpA, indicating multiple HGT events. The species found outside of their expected clusters might have adapted quickly to environmental changes as a result of HGT events, which accelerate the rate of adaption [46]. This is illustrated in the epsilon INK1197 order cluster; three of the four non-epsilon bacteria in that clade colonize humans either as pathogenic bacteria or as part of the intestinal microbiota A-1155463 order (see Figure 3 and Additional file 2: Table S3 for details). Conclusions The pldA gene in Helicobacter pylori has high nucleotide sequence identity due to purifying selection at the vast majority of residues. The result is a conserved H. pylori protein that likely has an evolutionarily stable function, although some probable interaction sites are subject to positive selection. Although HGT was detected by codon bias, GC content, and phylogenetic analysis, the biogeography of the pldA sequences indicated that the transfer was ancient. The protein structure of H. pylori OMPLA will yield a better understanding

of the positively selected sites, which may be surface-exposed regions. Our analyses indicated that pldA may be a niche-adapted protein; it was horizontally acquired, is highly conserved, but positive selection occurs at sites needed for possible pathogenic interactions. Methods Helicobacter pylori sample collection and pldA sequencing The pldA gene of 227 H. pylori isolates was sequenced. The samples included 207 Norwegian and Sepantronium nmr 20 Korean isolates. The Norwegian samples consisted of a total of 155 isolates from the Farnesyltransferase Sørreisa study [24] and 52 isolates collected from four hospitals in the Oslo region. Among these isolates, 40 had been previously described [33]. The Oslo isolates included samples with known foreign origins; four isolates with Indo-European

origins, two with Asian origins, and one with an African origin. DNA was isolated using BioRobot M48 and MagAttract DNA Mini M48 Kit (Qiagen Inc., Valencia, CA, USA). The pldA gene, including short parts of the up- and downstream genes, was amplified by polymerase chain reaction (PCR) with forward primer HP498/499-F (5’- ttatcgcgcctgtagtga -3’) and reverse primer HP499/500-R (5’- tatgatcgctggcatgga -3’) at an annealing temperature of 57°C. The 1068 base pair (bp) pldA-gene was sequenced using the ABI BigDye Terminators v 1.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA) with the PCR primers and the internal sequencing primers HP498/499-R (5’-ggttgatattggggtggta-3’), PLA-F (5’-tgtccaattcttggtatctc-3’), PLA-R (5’-atgcgataggtatagcctaag-3’) and HP499/500-F (5’-tatgatcgctggcatgga-3’). The sequencing products were analyzed with an ABI PRISM 3130 Genetic Analyzer (Applied Biosystems) and the sequences were aligned using Sequencher software (Gene Codes Corporation, Ann Arbor, MI, USA).

Diagnoses were established according to the WHO criteria [26] Wr

Diagnoses were established according to the WHO criteria [26]. Written informed consent was obtained from all patients in accordance with the Declaration of Helsinki and the ethical guidelines of the Charite University School of Medicine, which approved this study. DNA extraction Mononuclear cells from BM aspirates were isolated using Ficoll density gradient centrifugation as described [27]. DNA was extracted using Allprep DNA/RNA mini kit (Qiagen) as per the manufacturer’s SRT1720 cost instructions. ARMS analysis of IDH2-R140Q mutations All primers were designed using Primer

3 Software (Additional file 2: Table S2). ARMS analysis was performed using 2 control primers flanking exon 23 and 2 allele-specific primers IDH2-RI and IDH2-FI that are complementary to the wild-type (wt) and mutated alleles, respectively. To enhance specificity, both the primers had an additional

medium mismatch at the preliminary base. The PCR mixture and reaction conditions are specified in the Additional file 3: PCR reaction mixtures and conditions. The generated PCR products were analysed on a 1.5% agarose gel. Endonuclease restriction analysis of DNMT3A-R882H mutations PCR amplification for endonuclease restriction analysis was conducted using primers DNMT3A-ResF/R (Additional Ion Channel Ligand Library high throughput file 2: Table S2). PCR reaction mixture was prepared as that described for ARMS assay. The reaction conditions are specified in the Additional file 3. In all, 10 μl of the PCR product was directly applied for endonuclease treatment with 1 μl Fnu4HI and 5 μl of CutSmart Buffer (New England Biolabs). After incubation at 37°C for 15 min products were analysed on a 1.5% agarose gel containing 10% ethidium bromide (voltage 150 V). HRM assay The reaction mixture and HRM conditions are specified in the Additional file 3. The analysis was performed in a Rotor Gene 6000 Real-Time PCR Cycler (Qiagen). Samples, including a control sample for each mutation and wt allele, were analysed in duplicates.

For DNMT3A and IDH2, the wt allele was used Fossariinae for normalisation, while for IDH1 R132C mutation control was used as the baseline. Normalisation regions for the optimal detection of DNMT3A were 82°C-83°C (leading range) and 87°C-88°C (trailing range), for the optimal detection of IDH1 were 73°C-74°C (leading range) and 82°C-83°C (trailing range) and for the optimal detection of IDH2 were 77°C-78°C (leading range) and 87°C-88°C (trailing range). Confidence threshold was set to 70% for all the genes. DNA sequencing All the primers used for sequencing are listed in the Additional file 2: Table S2. All PCR reaction conditions are specified in the Additional file 3. The obtained products were purified using the PCR Purification Kit (Qiagen), as described in the manual. Sequencing reaction was performed using Big Dye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems). The sequencing products were purified using DyeEx 2.0 Spin Kit (Qiagen) according to the manufacturer’s instructions.

A repeat fluoroscopic contrast study of the drain showed resoluti

A repeat fluoroscopic contrast study of the drain showed resolution of the abscess PRIMA-1MET ic50 and fistula. The drain was then removed without complication. Three months following drain removal, the patient was noted to be tolerating a regular diet with no signs of infection or fistula drainage. She suffered only mild deconditioning and had no significant loss of IWR-1 supplier functional status. Figure 4 CT image of collapsed abscess cavity. CT image of the pelvis without contrast shows the drain in place and the abscess cavity completely collapsed. Discussion Migration of endoscopically placed biliary stents is a well recognized complication of ERCP. Less than 1% of migrated

stents cause intestinal perforation.[5] Of those that do perforate the bowel, the vast majority occur proximal to the ligament of Trietz (LOT). There have been a several case reports of intestinal perforation distal to the LOT, generally in the colon. [6–9] There have also been case reports describing small bowel perforation. [10–13] Generally speaking, a double pigtail stent (7F) is preferable in cases involving choledocholithiasis. A straight stent may migrate since there is nothing to hold it in place, even though there are side flaps. An exception might be an impacted stone that is tight on the stent and prevents migration. Dislodged straight stents are more likely to perforate

bowel whereas perforation with a pigtail is much more rare. Furthermore, straight 10 F plastic stents should generally be used for conditions such as strictures and tumours. The rationale this website for a double pigtail stent (7F) in this case is not known to the authors. Migrated stents causing complications have either been retrieved endoscopically or via laparotomy.[4, 7, 14] There is at least one documented case of a percutaneous intervention to remove a biliary stent causing a retroperitoneal duodenal perforation and bilioma. However, there has not been a documented case involving percutaneous methods to retrieve a migrated stent beyond the LOT. The

existing literature on this subject would advocate prompt and aggressive surgical intervention because of gross contamination, intraperitoneal abscess, Interleukin-3 receptor and bowel perforation.[4, 5] Prompt surgical intervention is generally indicated for small bowel perforations, especially in the setting of a highly contaminated field, bowel obstruction and generalized abdominal pain. Historically, bowel perforation from migrated bilary stents has been treated either by endoscopic retrieval or laparotomy should endoscopic means fail. There are reports in which endoscopy is used to retrieve stents and close bowel perforations via clip application, but this only applies to areas that are accessible to endoscopic instrumentation.[14] In our case, endoscopic means was not possible because the perforation was in the distal small bowel and associated with a partial small bowel obstruction.