Chronic pain following totally extra-peritoneal inguinal hernia repair: a randomized clinical trial comparing glue and absorbable tackers.

PURPOSE: Chronic pain following inguinal hernia repair occurs in up to 20% of patients. The underlying mechanism probably involves sensory nerve damage and abnormal healing that might be influenced by the materials chosen for mesh fixation. The main objective of this study was to compare glue and absorbable tackers on the rate of chronic pain after surgery in patients undergoing totally extraperitoneal inguinal hernia repair (TEP). METHODS: Patients undergoing (TEP) inguinal hernia repair were enrolled in a single-blind randomized clinical trial and were randomized for mesh fixation with glue (LIQUIBAND FIX 8 Neopharm) or absorbable tackers (SECURE STRAP Johnson & Johnson). Pain was assessed using a validated 4-point verbal-rank scale (none, mild, moderate, and severe) at 1 week, 1 month, 6 months, and 1 year postoperatively. Chronic pain was defined as pain persisting beyond 3 months. RESULTS: Two hundred and eight patients were analyzed. The groups were similar in age, gender, and hernia side. Chronic pain of any intensity was reported in 31.7% (66/208) after 6 months and in 13% (29/208) after 12 months. No differences in postoperative pain were observed between the two forms of mesh fixation. Still, when only those with severe pain were considered, mesh fixation with glue resulted in less pain compared to fixation by tackers (log-rank p = 0.025). At 1 year, 4 symptomatic recurrent hernias were identified in patients whose mesh was fixated with absorbable tackers. CONCLUSIONS: Patients who underwent TEP inguinal hernia repair with mesh fixated by glue suffered from less pain.

Gene Expression Pattern of Peyer's Patch Lymphocytes Exposed to Kagocel Suggests Pattern-Recognition Receptors Mediate Its Action.

Kagocel is a synthetic carboxymethylcellulose derivative copolymerized with gossypol. Clinical data evidence its safety and efficiency for the treatment of flu and other viral infections via enhancement of interferon production. The gut-associated lymphoid tissue seems a likely site of kagocel action. The study was aimed to investigate the molecular mechanisms of its action using murine Peyer's patches lymphocytes as a test system and the cytokines production and gene expression patterns as the primary outcomes. The Peyer's patches lymphocytes isolated from BALB/c mice were stimulated with concanavalin A, or, to mimic viral infection, with a combination of concanavalin A and TLR3 ligand poly I:C. After 24 h of stimulation the cells were treated with saline, 30, 100, or 300 µg/ml of kagocel, or, as positive controls, 300 µg/ml oats b-D-glucan or 300 µg/ml lentinan. After 24 and 72 h of incubation with these drugs cytokines production was analyzed with ELISA and gene expression pattern was investigated using nCounter Inflammation panel chips followed by bioinformatics analysis. Expression of genes involved in the inflammatory response, antiviral defense, lymphocytes survival and proliferation (C1qa, C2, C3, Ccl21a, Il11, Il1b, Il23a, Il5, Ltb4r2, Alox15, Pla2g4a, Ptger1, Mapkapk5, Hras, Ifna1, Tlr2, Mrc1, Mx2) was upregulated in kagocel-treated Peyer's patches lymphocytes. A list of plausible transcription factors (CEBPs, IRF, NFκB, RXR, Stat, Tead4, and ZSCAN) and master-regulators has been identified (cIAP, CIKS, dock9, MEKK1, FXR, IKK, IRAK, TRAF, dsRNA:TLR3:TRIF). The changes in gene expression pattern and the outcomes of bioinformatics analysis suggest that pattern recognition receptors, TLRs and dectin-1, are the key mediators of kagocel immunomodulatory action, with the possible involvement of interferon autocrine loop. The genes upregulated with kagocel include diverse components of the innate immune defense system.

Native Bovine Hydroxyapatite Powder, Demineralised Bone Matrix Powder, and Purified Bone Collagen Membranes Are Efficient in Repair of Critical-Sized Rat Calvarial Defects.

Here we evaluated the efficacy of bone repair using various native bovine biomaterials (refined hydroxyapatite (HA), demineralised bone matrix (DBM), and purified bone collagen (COLL)) as compared with commercially available bone mineral and bone autografts. We employed a conventional critical-sized (8 mm diameter) rat calvarial defect model (6-month-old male Sprague-Dawley rats, n = 72 in total). The artificial defect was repaired using HA, DBM, COLL, commercially available bone mineral powder, bone calvarial autograft, or remained unfilled (n = 12 animals per group). Rats were euthanised 4 or 12 weeks postimplantation (n = 6 per time point) with the subsequent examination to assess the extent, volume, area, and mineral density of the repaired tissue by means of microcomputed tomography and hematoxylin and eosin staining. Bovine HA and DBM powder exhibited excellent repair capability similar to the autografts and commercially available bone mineral powder while COLL showed higher bone repair rate. We suggest that HA and DBM powder obtained from bovine bone tissue can be equally applied for the repair of bone defects and demonstrate sufficient potential to be implemented into clinical studies.

Diagnostic Use of Endoscopic Retrograde Cholangiopancreatectography for Pancreatic Duct Injury in Trauma Patients.

BACKGROUND: Trauma patients diagnosed with pancreatic duct injury (PDI) have a high complication rate and prolonged hospital stay. The role of endoscopic retrograde cholangiopancreatography (ERCP) in the diagnosis of PDI remains unclear. During the last decade, our trauma unit incorporated ERCP into the management protocol for suspected PDI cases. OBJECTIVES: To determine whether ERCP is a sensitive tool to detect PDI. METHODS: This retrospective trauma patient series study assessed the diagnostic yield of ERCP in trauma cases with suspected PDI on computed tomography (CT) or intraoperatively. Between 1 January 2004 and 31 December 2011, 13 patients admitted to our medical center underwent ERCP for suspected PDI. Patient demographics, mechanism of injury, Injury Severity Score (ISS), time from injury to ERCP, and ERCP-related complications were documented and assessed. RESULTS: Of the 13 patients included in the analysis, 8 stable patients with suspected PDI on CT had no leak from the main pancreatic duct on ERCP. Two of them underwent surgery for suspected pancreatic transection. ERCP confirmed a main pancreatic duct leak in three patients. Two patients underwent ERCP for suspected PDI after "damage control" surgery. No leak from the pancreatic ducts was diagnosed. No pancreas-related complications or ERCP-related complications were observed. CONCLUSIONS: ERCP is a sensitive and relatively safe tool for the diagnosis of PDI, and its use might prevent unnecessary surgical interventions in selected trauma cases.

Reduced postoperative chronic pain after tension-free inguinal hernia repair using absorbable sutures: a single-blind randomized clinical trial.

BACKGROUND: Chronic pain after inguinal hernia repair occurs in 16% to 62% of patients. The underlying mechanism probably involves sensory nerve damage and abnormal healing that might be influenced by the materials chosen for the procedure. We hypothesize that nonabsorbable sutures used for mesh fixation to the surrounding tissues are associated with higher rates of chronic groin pain after surgery. STUDY DESIGN: We conducted a single-blind randomized clinical trial to compare the effect of absorbable braided sutures (Vycril; Ethicon) and nonabsorbable monofilament sutures (Prolene; Ethicon) used in inguinal hernia repair on the rate of chronic pain. We assessed chronic pain using a 4-point verbal-rank scale during a 1-year postoperative follow-up period. RESULTS: Study groups included 100 patients in each group. No age, sex, or hernia-side differences were observed between the study groups. Chronic pain rate after surgery was higher in the nonabsorbable monofilament suture group compared with the absorbable braided suture group (37 vs 26 patients; p = 0.056). Time to pain disappearance was longer in the nonabsorbable monofilament suture vs the absorbable braided suture group (115.3 days; 95% CI, 88-142.7 vs 77.4 days; 95% CI, 54.3-100.3; p = 0.038, respectively). A 1-year age increment reduces the risk for chronic pain occurrence by 2.2% (odds ratio = 0.978%; 95% CI, 0.961-0.995; p = 0.013). The use of nonabsorbable sutures increases the risk for chronic pain in 94.9% compared with absorbable sutures (odds ratio = 1.949; 95% CI, 1.039-3.658; p = 0.038). CONCLUSIONS: Nonabsorbable suture use in inguinal hernia repair is associated with a higher rate of chronic pain and a longer time to pain disappearance as compared with absorbable sutures.

Selection of patients with severe pelvic fracture for early angiography remains controversial.

BACKGROUND: Patients with severe pelvic fractures represent about 3% of all skeletal fractures. Hemodynamic compromise in unstable pelvic fractures is associated with arterial hemorrhage in less than 20% of patients. Angiography is an important tool in the management of severe pelvic injury, but indications and timing for its performance remain controversial. METHODS: Patients with major pelvic fractures [Pelvic Abbreviated Injury Score (AIS) >or= 3] admitted to two high volume Trauma Centers from January 2000 to June 2005 were identified and divided into two groups: Group I patients did not undergo angiography, Group II patients underwent angiography with/without embolization. Demographics, hemodynamic status on admission, concomitant injuries, Glasgow Coma Scale (GCS), Injury Severity Score (ISS), pelvic AIS, blood requirement before and after angiography, arterial blood gases and mortality were evaluated. Patients with an additional reason for hemodynamic instability were excluded. RESULTS: Charts of 106 patients were retrospectively reviewed. Twenty nine patients (27.4%) underwent angiography. Bleeding vessel embolization was performed in 20 (18.9%) patients. Patients who underwent angiography had a significantly higher pelvic AIS and a lower Base Excess level on admission. A blood transfusion rate of greater than 0.5 unit/hour was found to be a reliable indicator for early angiography. CONCLUSION: A high pelvic AIS, amount of blood transfusions and decreased BE level should be considered as an indicators for early angiography in patients with severe pelvic injury.

The properties of Bacillus cereus hemolysin II pores depend on environmental conditions.

Hemolysin II (HlyII), one of several cytolytic proteins encoded by the opportunistic human pathogen Bacillus cereus, is a member of the family of oligomeric beta-barrel pore-forming toxins. This work has studied the pore-forming properties of HlyII using a number of biochemical and biophysical approaches. According to electron microscopy, HlyII protein interacts with liposomes to form ordered heptamer-like macromolecular assemblies with an inner pore diameter of 1.5-2 nm and an outer diameter of 6-8 nm. This is consistent with inner pore diameter obtained from osmotic protection assay. According to the 3D model obtained, seven HlyII monomers might form a pore, the outer size of which has been estimated to be slightly larger than by the other method, with an inner diameter changing from 1 to 4 nm along the channel length. The hemolysis rate has been found to be temperature-dependent, with an explicit lag at lower temperatures. Temperature jump experiments have indicated the pore structures formed at 37 degrees C and 4 degrees C to be different. The channels formed by HlyII are anion-selective in lipid bilayers and show a rising conductance as the salt concentration increases. The results presented show for the first time that at high salt concentration HlyII pores demonstrate voltage-induced gating observed at low negative potentials. Taken together we have found that the membrane-binding properties of hemolysin II as well as the properties of its pores strongly depend on environmental conditions. The study of the properties together with structural modeling allows a better understanding of channel functioning.

Purification and cytotoxic properties of Bacillus cereus hemolysin II.

The hemolysin II from Bacillus cereus, HlyII, is a member of the beta-barrel pore-forming toxin family of secreted microbial proteins that includes the Staphylococcus aureus alpha-toxin. Compared with other proteins of the family, hemolysin II has 90 extra amino acids at its C-terminus. To examine more closely the cytotoxic and pore-forming properties of the protein, we have cloned and expressed it in Escherichia coli. We developed a purification procedure for the matured HlyII protein from both culture media and cell extracts using a combination of cation exchange and affinity chromatography together with gel-filtration. In both cases, the fully processed HlyII protein was purified as confirmed by N-terminal sequence analysis. The HlyII protein exhibits cytolytic activity of different extent on erythrocytes from various kinds of mammals. The results presented here show for the first time that two types of human cells are sensitive to HlyII action. In view of its broad cytotoxic activity as well as the ability to interact with artificial membranes, we assume that HlyII needs no specific receptor to bind to cell membranes.

Limits of diagnostic accuracy of anti-hepatitis C virus antibodies detection by ELISA and immunoblot assay.

When human sera samples are tested for anti-hepatitis C virus (HCV) antibodies using different ELISA kits as well as immunoblot assay kits discrepant results often occur. As a result the diagnostics of HCV infection in such sera remains unclear. The purpose of this investigation is to define the limits of HCV serodiagnostics. Overall 7 different test kits of domestic and foreign manufacturers were used for the sampled sera testing. Preliminary comparative study, using seroconversion panels PHV905, PHV907, PHV908 was performed and reference kit was chosen (Murex anti-HCV version 4) as the most sensitive kit on the base of this study results. Overall 1640 sera samples have been screened using different anti-HCV ELISA kits and 667 of them gave discrepant results in at least two kits. These sera were then tested using three anti-HCV ELISA kits (first set of 377 samples) or four anti-HCV ELISA kits (second set of 290 samples) at the conditions of reference laboratory. In the first set 17.2% samples remained discrepant and in the second set - 13.4%. "Discrepant" sera were further tested in RIBA 3.0 and INNO-LIA immunoblot confirmatory assays, but approximately 5-7% of them remained undetermined after all the tests. For the samples with signal-to-cutoff ratio higher than 3.0 high rate of result consistency by reference, ELISA routing and INNO-LIA immunoblot assay was observed. On the other hand the results of tests 27 "problematic" sera in RIBA 3.0 and INNO-LIA were consistent only in 55.5% cases. Analysis of the antigen spectrum reactive with antibodies in "problematic" sera, demonstrated predominance of Core, NS3 and NS4 antigens for sera, positive in RIBA 3.0 and Core and NS3 antigens for sera, positive in INNO-LIA. To overcome the problem of undetermined sera, methods based on other principles, as well as alternative criteria of HCV infection diagnostics are discussed.

Age-Related Changes in Population of Stromal Precursor Cells in Hematopoietic and Lymphoid Organs.

It is shown that the content of precursor cells of stromal tissue (CFC-F) in the hemopoietic and lymphoid organs of SAMP (rapidly-ageing mice) and SAMR mice (mice with a normal ageing rate) decreases as the animals grow older. However the decrease in the content of CFC-F in SAMP mice begins substantially earlier - in the age group of 9-11 months, while in the SAMR mice - only in the age group of 16-19 months. It was found that the age reduction of the number to an equal degree relates to the whole population of CFC-F, in particular both the fraction of weakly-linked CFC-F, which is isolated by means of mechanical disaggregation of the tissue, and the fraction which may only be isolated using trypsin. It is shown that the concentration of inducible osteogenic precursor cells (IOPC) in the spleen of guinea pigs does not change with age, but their content in that organ in old animals (2-3 years old) drops by two times. It was found that in elderly animals the mass of the ectopic osseous tissue, formed by the implantation of an osteoinductor (autologous epithelium of the urinary bladder) in a system open for entrance of cells, decreases by two times. After curettage of the medullary cavity of guinea pig tibia (i.e. under conditions of an increased demand for osteogenic cells) the mass of induced ectopic osseous tissue decreases by 4 times, which indicates to the possible functional relationship between the pool of determined and inducible osteogenic precursor cells. On the whole, the obtained data show that during ageing there is a reduction in the number of stromal precursor cells (CFC-F and IOPC), which form a specific microenvironment for hemopoietic and lymphoid organs, which is important to understand the role of these cells in the development of age pathologies, in particular senile osteoporosis.