Laparoscopic Extravesical Reimplantation in Children with Primary Obstructive Megaureter.

Introduction: Conservative management of primary obstructive megaureter (POM) appears as the best option in patients with adequate ureteral drainage. Nevertheless, surgical intervention is indicated in cases of recurrent urinary tract Infections (UTIs), deterioration of split renal function, and significant obstruction. The gold standard includes: Ureteral reimplantation with or without tapering by open approach. Our objective is to report our results in the treatment of POM by Laparoscopic-Assisted Extracorporeal Ureteral Tapering Repair (EUTR) and Laparoscopic Ureteral Extravesical Reimplantation (LUER) and to evaluate the efficacy and security of this procedure. Materials and Methods: From January 2011 to January 2018 a retrospective study was carried out by reviewing the clinical records of 26 patients diagnosed with POM. All patients underwent laparoscopic ureteral reimplantation following Lich Gregoir technique. In cases of ureteral tapering, an EUTR was performed with Hendren technique. Results: In all patients LUER and EUTR were performed without conversion. No ureteral tapering was necessary in six patients. There were no intraoperative complications. At 3 months in postoperative, 1 patient presented a febrile UTI, and subsequently, a vesicoureteral reflux (VUR) grade III was diagnosed by voiding cystourethrogram. In this case, a redo laparoscopic surgery was performed. After long-term follow-up, all patients were asymptomatic without recurrence of POM or VUR. Conclusion: Laparoscopic-assisted EUTR and LUER following Lich Gregoir technique for POM constitutes a safe and effective option, with a success rate similar to that of open procedure. Nevertheless, larger randomized prospective trials and long-term follow-up are required to validate this technique.

Pediatric kidney retransplantation focused on surgical outcomes.

INTRODUCTION: Despite survival rates after pediatric kidney transplantation (KT) are on the rise it is still likely that most pediatric recipients will require more than one retransplant in their lifetime. The earlier the age at the first KT the higher is the risk of repeat pediatric kidney transplantation (RPKT). OBJECTIVE: The current study aims to analyze the outcomes of repeat pediatric kidney transplantation (RPKT) among pediatric kidney transplant recipients focusing on surgical complications and compare the outcomes of second and subsequent grafts with those of the first kidney graft. MATERIALS AND METHODS: Retrospective study of RPKT (<18 years) undertaken between January 2000-2020. We analyzed primary etiology of renal disease, time to graft loss (GL), etiology of initial graft failure, history of acute rejection, previous delayed graft function, HLA-mismatches at the initial transplant, surgical complications and outcomes. Additionally, we compared the characteristics and outcomes of patients who underwent RPKT (group 1) with those who received a first kidney graft (group 2). RESULTS: Out of 229 kT, 59 patients underwent RPKT (26 females/33 males). At the time of RPKT median age was 11.37 years (SD:5.7). The most frequent primary renal disease was congenital nephrotic syndrome in 11 (18.6%). Fifty-four (91.5%) were on renal replacement therapy at the time of transplant. Fourty-one patients received their second KT (69.5%), 14 (23.7%) the third, 3 (5.1%) the fourth and 1 (1.7%) the fifth. Transplant graft nephrectomy (GN) was performed in 26 patients (44.1%) prior to retransplantation. Fifty-four (91.5%) received a cadaveric graft and 5 (8.5%) a living-related graft. An extraperitoneal approach was achieved in 53 patients (89.8%), whereas in the remaining 6 (10.2%) the graft was placed intraperitoneally. We observed 10 surgical complications (16.9%): 9 major which required reintervention and 1 minor (perirenal hematoma). No vascular complications were observed and none of the surgical complications were involved in graft loss. Graft survival at 1,3 and 5 years was 91%, 84% and 73% respectively. The most frequent cause of GL was chronic graft nephropathy in 15 (25.4%). After a mean follow-up of 9.40 years (SD: 4.7) only 2 patients died (3.4%), both with functioning grafts. DISCUSSION: Pediatric recipients of second and subsequent kidney grafts constitute a remarkable high-risk population but are becoming more frequent at reference pediatric transplant centers. CONCLUSIONS: RPKT is technically challenging but can yield good results. In our series overall the incidence of surgical complications and particularly vascular complications was low.

Pediatric laparoscopic mitrofanoff procedure- preliminary results of a simplified technique.

INTRODUCTION: Laparoscopic appendicovesicostomy (LA) is a rather new technique and still a challenging procedure even for the most experienced surgeons because it requires advanced laparoscopic experience and surgical skill. The aim of this study is to analyze the short-term results and benefits of laparoscopic LA in children. MATERIALS AND METHODS: Prospective study of children undergoing LA at our institution between January 2018 and October 2021. The procedure was perfomed using a laparoscopic transperitoneal approach. The distal end of the appendix was spatulated and reimplanted in the bladder by a modified Shanfield technique. The proximal end was brought out as the cutaneous umbilical stoma. RESULTS: Over the study period 15 patients underwent LA (14 males,1 female). Mean age at intervention was 8.8 years (SD:3.1). Indication for surgery was pain during CIC in 13 (86.7%) and difficulty for CIC in 2 (13.3%). Eight patients (53.5%) presented end-stage renal disease (ESRD). Median operative time was 217.3 min (r:140-300). Two patients (13.3%) experienced early postoperative complications: ileus (1) and internal hernia over the mesoappendix with subsequent intestinal obstruction. Mean hospital stay was 6.8 days (SD:1.7). Four patients (26.7%) experienced late postoperative stoma related complications: stomal stenosis (1), granuloma (1) and inability to catheterize (2). With a mean follow-up of 21.46 months (SD: 13) all except the patent who lost the conduit are continent and on CIC every 3 h. CONCLUSIONS: We suggest that LA by this technique is effective, safe and reproducible, and is associated with good short-term results. The complication rate is similar to the open procedure being intestinal obstruction due to internal hernia probably the most serious.

Laparoscopic extravesical ureteral reimplantation (LEUR) of obstructed megaureters: STEP-BY-STEP video of a simplified approach.

INTRODUCTION: In this video we aim to describe step-by-step a simplified surgical technique for the treatment of primary and secondary obstructed megaureters in children by laparoscopic extravesical ureteral reimplantation (LEUR). MATERIAL AND METHODS: A transperitoneal approach is used in all cases. The distal ureter is dissected and transected at the level of the stenosis. Detrusor fibers are divided to expose bladder mucosa and the distal ureter is introduced into the bladder to create a valve-like mechanism (modified-Shanfield technique). RESULTS: Between 2016 and 2020, 9 patients underwent LEUR (5 female, 4 male). Mean age at surgery was 31.6 months (SD:22.74) and indications were infection (2), obstruction (3), increase hydronephrosis (2) and decrease in renal differential function (2). The procedure was completed laparoscopically in all cases with a median operative time of 144 min (r: 120-160). The postoperative MAG-3 renogram revealed a non-obstructive pattern in 8/9 patients and MCUG demonstrated absence of VUR in all 8/9. With a mean follow-up of 2.4 years (SD:1.4) all are asymptomatic. CONCLUSIONS: LEUR by this new simplified technique is a feasible treatment of megaureters associated with good short and long-term results.

Laparoscopic extravesical ureteral reimplantation for correction of primary and secondary megaureters: Preliminary report of a new simplified technique.

PURPOSE: To describe a simplified surgical technique for the treatment of primary and secondary obstructed megaureters in children by laparoscopic extravesical ureteral reimplantation (LEUR) and evaluate the short-term outcomes. METHODS: Prospective study of children with primary and secondary megaureters treated at our institution between 2016 and 2018 by LEUR. A transperitoneal approach was used in all cases. The distal ureter was transected at the level of the stenosis. Detrusor muscle fibers were divided to expose bladder mucosa. The distal ureter was introduced into the bladder to create a valve-like mechanism. The ureter was fixed to the bladder mucosa by four stitches. We analyzed indications for surgery, complications and outcomes. Definition of success was relieved of obstruction and absence of VUR. RESULTS: Six patients with a mean age of 28.83 months (SD: 21.4) underwent LEUR. Indications for surgery were: infection [2], obstruction [2], decrease in renal differential function [1] and increase in hydronephrosis [1]. There were no intraoperative complications. Resolution of hydronephrosis, obstruction and VUR was achieved in all cases. Mean follow-up was 13 months (SD: 6.67). CONCLUSION: LEUR for the correction of primary and secondary megaureters by this new technique is safe. The most important advantage is that this technique is simpler and more easily reproducible than conventional LEUR. However, long-term follow-up is required. TYPE OF STUDY: Prospective, observational. LEVEL OF EVIDENCE: Level III.

Hemorrhagic cystitis after hematopoietic stem cell transplantation: A challenge for the pediatric urologist.

OBJECTIVE: Hemorrhagic cystitis (HC) is a serious event that can occur after hematopoietic stem cell transplantation (HSCT). Treatment goals are primarily to preserve life, and then the functionality of the bladder. There is no standard therapeutic approach for HC. Described treatment options provide low success rates and are related to potential life-threatening side effects. The aim of this study was to describe our experience in treatment of HC following HSCT. PATIENTS AND METHODS: This was a retrospective study of patients with HC treated at our institution between January 2010 and October 2016. We analyzed demographics, underlying diagnosis, and treatment modalities. RESULTS: We treated 39 patients with HC. Mean age was 9.4 years (SD 4.20) and 64% were males. Acute leukemia was the most common underlying diagnosis in 27 (69%). Mean time from HSCT to HC onset was 55.46 days (SD 112.35). HC grades were: I (3), II (21), III (8), and IV (7). BK-viuria was present in 34 patients (87.2%). Non-invasive treatment was performed in 28 patients (71.8%). The remaining 11 (28.2%) required urological intervention (all high-grade), consisting of bladder irrigation in all of these. Additional treatments consisted of: intravesical cidofovir (4), intravesical sodium hyaluronate (5), cystoscopy and clot evacuation (4), selective angioembolization (2), percutaneous nephrostomy (1), and open extraction of bladder clots and cutaneous cystotomy (1). Overall, eight patients (20.5%) died as a result of the malignancy (3 in the urological intervention group), and of these four had active HC at death. Mean follow-up was 36.2 months (SD 24.9). CONCLUSION: HC is associated with high morbidity and mortality. Treatment should be individualized and designed to prioritize survival. However, bladder function should be preserved for the future.

Vascular thrombosis in pediatric kidney transplantation: Graft survival is possible with adequate management.

OBJECTIVE: Vascular thrombosis (VT) in pediatric kidney transplantation (KT) is a dreaded event that leads to graft loss in almost 100% of cases. In recent years, VT has become the most common cause of early graft loss. The aim of this study was to analyze our experience in diagnosis and treatment of VT and the impact of a new management protocol on patient outcome. METHODS: We conducted a retrospective study of 176 consecutive KT performed at our institution by the pediatric urology team between January 2000 and December 2015 and identified patients with VT. A protocol of prevention and early detection of VT was introduced in 2012. RESULTS: Out of 176 KT, nine cases of VT were identified (5.1%). The mean recipient age was 5.1 years (SD 4.9 years) and mean weight was 22.28 kg (SD 15.6 kg). Diagnosis was intraoperative in two cases and within the first 24 h after surgery in the remaining seven. Immediate surgical exploration was performed after diagnosis in all. Of the five episodes that occurred before 2012, all developed complete graft ischemia requiring transplantectomy. However, in the four cases diagnosed after 2012, graft perfusion could be restored in three, and abdominal wall closure with a mesh and delayed sequentially closure under ultrasound guidance was performed. With a follow-up of 30, 25, and 20 months, the three recovered grafts are still functioning normally. CONCLUSIONS: Increased awareness and the application of a protocol for prevention, detection and treatment of VT in pediatric KT can prevent graft loss. Immediate surgical intervention is mandatory after diagnosis. Avoiding compartment syndrome with delayed sequential closure may be useful to improve graft survival.

Laparoscopic-Assisted Extravesical Ureteral Reimplantation and Extracorporeal Ureteral Tapering Repair for Primary Obstructive Megaureter in Children.

BACKGROUND: Open surgery is a preferred treatment for primary obstructive megaureter (POM) in cases where the conservative treatment fails, with reported success rates of 90%-96%. OBJECTIVE: To describe our initial experience in the treatment of POM by laparoscopic-assisted extracorporeal ureteral tapering repair (EUTR) and laparoscopic ureteral extravesical reimplantation (LUER) by following Lich Gregoir technique as an alternative to open surgery. DESIGN, SETTING, AND PARTICIPANTS: A total of 7 patients with POM underwent laparoscopic-assisted extracorporeal ureteral tapering repair and ureteral extravesical reimplantation by following Lich Gregoir technique between 2011 and 2014. Postoperative follow-up included the following: Renal and bladder ultrasound, voiding cystourethrogram (VCUG), and mercaptoacetyltriglycine (MAG3) renogram were done at 6 months. Outcome Measurements and Statistical Analysis: Statistical analysis was performed by using the SPSS software package (version 15.0; SPSS, Chicago, IL), and P < .05 was considered statistically significant. Paired tests and Wilcoxon test were performed to compare pre- and post-measures. RESULTS: LUER and EUTR were completed successfully in all patients without conversion. A postoperative MAG3 renogram showed nonobstructive pattern in all patients. Statistical analysis revealed significant differences before and after surgery in the average time of elimination on the MAG3 renogram (T½ 59.10 minutes versus 13.57 minutes, P < .0001). After medium-term follow-up, the overall POM resolution was 100%. One case of vesicoureteral reflux (VUR) was found during VCUG control. A total of 7 patients were asymptomatic without recurrence of POM. CONCLUSION: Laparoscopic-assisted extracorporeal ureteral tapering repair and ureteral extravesical reimplantation by following Lich Gregoir technique for POM constitutes a safe and good option when the first line of treatment fails, with a success rate similar to the open procedure. Nevertheless, larger randomized prospective trials and long-term follow-up are required to validate this technique.

Evaluation of the Sepsis Flow Chip assay for the diagnosis of blood infections.

BACKGROUND: Blood infections are serious complex conditions that generally require rapid diagnosis and treatment. The big challenge is to reduce the time necessary to make a diagnosis with current clinical microbiological methods so as to improve the treatment given to patients. METHODS: In this study, we assess for the first time the Sepsis Flow Chip assay, which is a novel diagnostic assay for simultaneous rapid-detection of the vast majority of bloodstream pathogens, including Gram-positive and Gram-negative bacteria and fungi, in the same assay, and for the detection of most common antibiotic resistance genes. The SFC assay is based on multiplex PCR and low density DNA arrays. RESULTS: Positive blood cultures from 202 consecutive bacteremia patients were analyzed by SFC assay and the results were compared with the results obtained by the gold standard methodology used in clinical microbiology diagnostic laboratories (EUCAST guidelines). SFC assay overall sensitivity and specificity for bacterial identification were 93.3% and 100% respectively and sensitivity and specificity for the identification of antibiotic genetic resistance determinants were 93.6% and 100% respectively. CONCLUSIONS: This is the first evaluation of SFC assay in clinical samples. This new method appears to be very promising by combining the high number of distinct pathogens and genetic resistance determinants identified in a single assay. Further investigations should be done to evaluate the usefulness of this assay in combination with clinical multidisciplinary groups (stewardship), in order for the results to be applied appropriately to the management of patients`infectious processes.

Kidney transplantation in children weighing 15 kg or less is challenging but associated with good outcome.

OBJECTIVE: Pediatric kidney transplantation (KT) in small children is assumed to be related to potential surgical complications that may cause severe morbidity and graft loss. The aim of our study was to analyze the outcome of KT recipients weighing ≤15 kg, focusing on surgical complications, associated morbidity and mortality, as well as allograft loss. METHODS: We reviewed our retrospective institutional database for recipients of KT between January 2000 and December 2014 with body weight ≤15 kg. RESULTS: Forty-four children weighing ≤15 kg, out of a total of 164 children (26.8%), received a deceased donor KT at our center during the study period. Mean weight was 10.10 ± 2.9 kg (3-15 kg), and weight was ≤10 kg in 23 patients (52.3%). The allograft was implanted intraperitoneally in two cases (4.5%) and extraperitoneally in the remaining 42 (95.5%). Two patients received a simultaneous double liver-kidney transplant. Postoperative complications appeared in 10 patients (22.7%) and eight required reintervention. Five allografts (11.4%) were lost secondary to surgical complications. No statistically significant differences in surgical complications were observed when compared with patients weighing >15 kg. Actuarial graft survival was 81% and 73% at 1 and 5 years, respectively. No significant differences in graft survival were observed compared with patients >15 kg. Mean follow-up was 84.95 ± 50 months (1-190 months). CONCLUSIONS: Our results demonstrate that KT in children weighing ≤15 kg is challenging but not associated with increased risk of surgical complications or early graft loss.

Streptococcus gallolyticus infection in colorectal cancer and association with biological and clinical factors.

There is an unambiguous association of Streptococcus gallolyticus infection with colorectal cancer, although there is limited information about epidemiology or interaction between molecular and environmental factors. We performed an original quantitative analysis of S. gallolyticus in unselected colorectal cancer patients (n = 190) and their association with clinical, pathological tumor molecular profiles (microsatellite instability, hypermethylator phenotype and chromosomal instability pathways), and other biological factors in colorectal tumor and normal tissues (cytomegalovirus and Epstein-Barr virus infection). We developed a new quantitative method to assess bacterial load. Analytical validation was reached with a very high sensitivity and specificity. Our results showed a 3.2% prevalence of S. gallolyticus infection in our unselected cohort of colorectal cancer cases (6/190). The average S. gallolyticus copy number was 7,018 (range 44-34,585). No previous reports relating to S. gallolyticus infection have been published for unselected cohorts of patients. Finally, and despite a low prevalence of S. gallolyticus in this study, we were able to define a specific association with tumor tissue (p = 0.03) and with coinfection with Epstein-Barr virus (p = 0.042; OR: 9.49; 95% IC: 1.1-82.9). The prevalence data provided will be very useful in the design of future studies, and will make it possible to estimate the sample size needed to assess precise objectives. In conclusion, our results show a low prevalence of S. gallolyticus infection in unselected colorectal cancer patients and an association of positive S. gallolyticus infection with tumor tissue and Epstein-Barr virus coinfection. Further studies will be needed to definitively assess the prevalence of S. gallolyticus in colorectal cancer and the associated clinicopathological and molecular profiles.

Treatment of Post-transplant Lymphocele in Children.

OBJECTIVE: To review our single-center experience in managing posttransplant lymphoceles in pediatric kidney recipients. Lymphoceles are well-known complications after pediatric kidney transplantation (KT). However, there is no standard treatment for lymphoceles, and the literature lacks consensus on which is the most appropriate approach. MATERIALS AND METHODS: We reviewed our retrospective institutional database for recipients of pediatric KT performed between January 2000 and December 2015 who developed lymphoceles. RESULTS: Out of the 176 patients who underwent KT, lymphoceles occurred in 9 (5.1%) patients. The mean age of recipients in this group was 12.8 years (standard deviation [SD] 4.8) (r: 1-17) and the mean body weight was 43.1 kg (SD 18.8) (r: 9.5-69). Mean lymphocele onset was 32.2 days (SD 23.4) (r: 11-85) post transplantation. Six patients presented with increased serum creatinine from the baseline, whereas 3 patients remained asymptomatic. Ultrasound was the primary diagnostic procedure in all patients. Lymphoceles resolved spontaneously in asymptomatic patients (n = 3), and thus these patients were not further treated. All symptomatic patients (n = 6) were treated: 2 underwent percutaneous catheter drainage and 4 underwent transcatheter sclerotherapy (TS). The main sclerosing agent used was povidone-iodine. In 3 patients, TS with povidone-iodine failed, and they underwent additional procedures: 2 underwent TS with polidocanol and 1 underwent open drainage. There was no graft loss in any of the patients, and no recurrence was documented during a follow-up period of mean 30.3 months (SD 15.6) (r: 7-57). CONCLUSION: There is no gold-standard treatment for lymphoceles in children, and reports in the literature on the topic are scarce. Percutaneous catheter drainage with or without TS is safe and effective, although it can lengthen hospitalization and increase morbidity.

Evaluation of the Initial Treatment of Ureteroceles.

OBJECTIVE: To assess the results of the initial therapeutic approach to ureteroceles at our institution and the need for further interventions. PATIENTS AND METHODS: This is a retrospective study of all pediatric cases of ureterocele diagnosed at our center between January 2000 and December 2011. RESULTS: Forty-three patients were analyzed. Initial diagnoses were ureterohydronephrosis in 34 (33 prenatal), febrile urinary tract infection in 5, and prolapsed ureterocele in 3. Expectant management was decided upon in 6 patients (14%). Of these, 1 required surgery. The remaining 37 (86%) initially underwent surgery: transurethral puncture (18), heminephrectomy (14), nephrectomy (3), and reimplantation (2). Twelve (66.6%) of the 18 patients who underwent primary puncture progressed well and required no further intervention. New-onset vesicoureteral reflux to the upper pole appeared after puncture in 3 patients, but none required treatment. Only 6 patients (33.3%) underwent a second procedure. Mean follow-up was 8.5 years (standard deviation: 3.08). CONCLUSION: Early endoscopic puncture is useful for decompression and often the definitive treatment. Although new-onset vesicoureteral reflux into the punctured system is the most common complication, it often resolves spontaneously. Early heminephrectomy in patients with nonfunctioning upper moieties yields excellent results but may not be necessary. Some patients may not need transurethral puncture or any surgical intervention at all.

Analysis of microbiota in stable patients with chronic obstructive pulmonary disease.

To identify the bacterial diversity (microbiota) in expectorated sputum, a pyrosequencing method that investigates complex microbial communities of expectorated sputum was done in 19 stable chronic obstructive pulmonary disease patients (mean (SD) FEV1: 47 (18%) of predicted value). Using conventional culture, 3 phyla and 20 bacterial genera were identified, whereas the pyrosequencing approach detected 9 phyla and 43 genera (p < 0.001). In sputum the prevalent genera with pyrosequencing approach were Streptococcus, Actinomyces, Neisseria, Haemophilus, Rothia, Fusobacterium, Gemella, Granulicatella, Porphyromonas, Prevotella and Veillonella. Enterobacteriaceae, detected frequently in conventional culture, were not significantly detected with pyrosequencing methods. In addition, we found that important pathogens such as Haemophilus and Moraxella were detected more frequently with the new genetic procedures. The presence of Enterobacteriaceae is probably overestimated with conventional culture, whereas other difficult cultivable pathogens are underestimated. These studies open a new perspective for evaluating the role of bacterial colonization in chronic obstructive pulmonary disease pathogenesis and progression.

Failed pyeloplasty in children: Is robot-assisted laparoscopic reoperative repair feasible?

OBJECTIVE: In this study we aim to demonstrate that robot-assisted laparoscopic (RAL) reoperative repair is safe and effective and even less technically demanding than open repair for recurrent ureteropelvic-junction obstruction (UPJO). STUDY DESIGN: A retrospective study was conducted of all cases of failed open pyeloplasties who underwent RAL reoperative repair at our institution between January 2010 and December 2013. The general surgical procedure was the same we previously described for robot-assisted laparoscopic pyeloplasty. Success was defined as: improvement in the degree of hydronephrosis at ultrasound, improvement of diuretic washout time at postoperative diuretic renogram (<15 min), improvement or at least stable differential renal function and absence of symptoms. These radiographic and symptomatic criteria of success were considered the primary outcomes. Secondary outcomes included complications and length of hospital stay. RESULTS: Between 2000 and 2013 a total of 153 patients underwent open Anderson-Hynes dismembered pyeloplasty. Of these 9 (6%) had recurrent UPJO. Four patients underwent open redo pyeloplasty. As a result, our study population comprised 5 children who underwent reoperative RALP repair. Patient characteristics and outcomes are summarized in the table below. Our success rate was 100%. DISCUSSION: Due to the low failure rate of open dismembered pyeloplasty there is no consensus on the best surgical approach for recurrent obstruction. While endoscopic approaches have been favored in adults, children have shown better success rates with repeat pyeloplasty. Laparoscopic salvage pyeloplasty for failed open procedures has become more popular and has been shown to result in excellent outcomes while providing the advantages of minimally invasive surgery. To date, the literature regarding the use of RALP for failed open procedures in the pediatric population is scarce. Only 2 pediatric series of robotic reoperative pyeloplasty have been reported by Helmal et al. (9 patients) and Lindgren et al. (16 patients) with a success rate of 100 and 88%, respectively. Although this is one of the first published studies about robot-assisted laparoscopic reoperative repair for failed open pyeloplasty in pediatric patients, we acknowledge the limitations of our study due to the small number of patients, its retrospective nature and limited follow-up time. CONCLUSIONS: The incidence of failed open pyeloplasty is as low as 5% and management remains controversial. As reported by other authors, we believe that crossing vessels play a particularly important role in secondary obstruction and adversely impact the outcome. Redo pyeloplasty, open or minimally invasive, is associated with high success rates (80-100%) and therefore considered the treatment of choice by the majority of authors nowadays. Additionally, RALP for secondary procedures has demonstrated to be safe and even less technically demanding when compared to the open approach, providing the advantages of minimally invasive surgery.

Relationship between tobacco, cagA and vacA i1 virulence factors and bacterial load in patients infected by Helicobacter pylori.

BACKGROUND AND AIM: Several biological and epidemiological studies support a relationship between smoking and Helicobacter pylori (H. pylori) to increase the risk of pathology. However, there have been few studies on the potential synergistic association between specific cagA and vacA virulence factors and smoking in patients infected by Helicobacter pylori. We studied the relationship between smoking and cagA, vacA i1 virulence factors and bacterial load in H. pylori infected patients. METHODS: Biopsies of the gastric corpus and antrum from 155 consecutive patients in whom there was clinical suspicion of infection by H. pylori were processed. In 106 patients H. pylori infection was detected. Molecular methods were used to quantify the number of microorganisms and presence of cagA and vacA i1 genes. A standardized questionnaire was used to obtain patients' clinical data and lifestyle variables, including tobacco and alcohol consumption. Adjusted Odds Ratios (ORadjusted) were estimated by unconditional logistic regression. RESULTS: cagA was significantly associated with active-smoking at endoscope: ORadjusted 4.52. Evidence of association was found for vacA i1 (ORadjusted 3.15). Bacterial load was higher in active-smokers, although these differences did not yield statistical significance (median of 262.2 versus 79.4 copies of H. pylori per cell). CONCLUSIONS: The association between smoking and a higher risk of being infected by a virulent bacterial population and with higher bacterial load, support a complex interaction between H. pylori infection and environmental factors.

[Characterization of gene expression associated with both the AcrAB/TolC system and the membrane permeability, in Salmonella spp isolates with and without gyrA mutation]. / Caracterización de la expresión de genes asociados con el sistema AcrAB/TolC y la permeabilidad de membrana en aislados de Salmonella spp. con y sin mutación en gyrA.

Introduction. The marA, soxS, ramA, acrB and ompF genes have been studied in order to characterize mechanisms of AcrAB-TolC active efflux pumps and membrane permeability alterations that reduce fluoroquinolones susceptibility in Salmonella spp. Methods. Mutations in marA, soxS, ramA, acrB and ompF genes were detected, as well as their expression levels in presence and absence of ciprofloxacin, calculating the level of change between them by qPCR. Data were analysed by using SPSS 19.0. Results. No mutations in these genes were found, but both AcrAB-TolC regulatory genes and structural acrB gene expression were affected by ciprofloxacin in both mutant strains and wild type bacterial strains (WT). The activation of the marA gene in presence of drug was higher in WT strains (level of change 0.823) than in mutants strains (level of change 0.158; p=0.049). In gyrA mutants, a reduction in ompF gene expression in presence of ciprofloxacin was found (level of change -0.949 p=0.017). Conclusion. The reduction of fluoroquinolones susceptibility in Salmonella spp is a complex process, in which several different bacterial mechanisms are involved. This study has found a high difference in the degree of participation among studied mechanisms, between bacterial strains with and without gyrA mutation. Whereas WT strains activated efflux pumps especially through marA gene, mutants supressed ompF gene expression related to porins.

Caracterización de la expresión de genes asociados con el sistema AcrAB/TolC y la permeabilidad de membrana en aislados de Salmonella spp. con y sin mutación en gyrA / Characterization of gene expression associated with both the AcrAB/TolC system and the membrane permeability, in Salmonella spp isolates with and without gyrA mutation

Introducción. Se han estudiado los genes marA, soxS, ramA, acrB y ompF para caracterizar los mecanismos del sistema de expulsión activa AcrAB/TolC y las alteraciones en la permeabilidad de membrana que reducen la sensibilidad a fluoroquinolonas en Salmonella spp. Métodos. Se detectaron las mutaciones en los genes marA, soxS, ramA, acrB y ompF y se cuantificó su nivel de expresión en presencia y ausencia de ciprofloxacino calculando su valor de nivel de cambio por qPCR. Los datos se analizaron estadísticamente mediante el programa SPSS 19.0. Resultados. No se encontraron mutaciones en ninguno de los genes, pero la expresión de los genes reguladores de AcrAB/TolC y del gen estructural acrB se vieron afectados por la presencia de ciprofloxacino tanto en las cepas con mutación en gyrA como en las cepas silvestres. La activación del gen marA en presencia del fármaco fue mayor en las cepas silvestres (nivel de cambio de 0,823) que en las mutantes (nivel de cambio de 0,158; p=0,049). Se vio una disminución de la expresión del gen ompF en presencia de ciprofloxacino en cepas con mutación (nivel de cambio de -0,949 p=0,017). Conclusión. La disminución de la sensibilidad a fluoroquinolonas en Salmonella spp es un proceso complejo, donde intervienen diferentes mecanismos bacterianos. Este estudio encuentra gran diferencia en el grado de participación de los mecanismos estudiados entre las cepas con y sin mutación en gyrA. Mientras que las cepas silvestres activan el sistema de expulsión activa, especialmente a través del gen marA, las cepas con mutación reprimen la expresión del gen ompF, relacionado con las porinas (AU)

Introduction. The marA, soxS, ramA, acrB and ompF genes have been studied in order to characterize mechanisms of AcrAB-TolC active efflux pumps and membrane permeability alterations that reduce fluoroquinolones susceptibility in Salmonella spp. Methods. Mutations in marA, soxS, ramA, acrB and ompF genes were detected, as well as their expression levels in presence and absence of ciprofloxacin, calculating the level of change between them by qPCR. Data were analysed by using SPSS 19.0. Results. No mutations in these genes were found, but both AcrAB-TolC regulatory genes and structural acrB gene expression were affected by ciprofloxacin in both mutant strains and wild type bacterial strains (WT). The activation of the marA gene in presence of drug was higher in WT strains (level of change 0.823) than in mutants strains (level of change 0.158; p=0.049). In gyrA mutants, a reduction in ompF gene expression in presence of ciprofloxacin was found (level of change -0.949 p=0.017). Conclusion. The reduction of fluoroquinolones susceptibility in Salmonella spp is a complex process, in which several different bacterial mechanisms are involved. This study has found a high difference in the degree of participation among studied mechanisms, between bacterial strains with and without gyrA mutation. Whereas WT strains activated efflux pumps especially through marA gene, mutants supressed ompF gene expression related to porins (AU)

Frecuencia de la resistencia a fluoroquinolonas en Mycobacterium tuberculosisen Elche (Alicante) / Frequency of fluoroquinolone resistance in Mycobacterium tuberculosis in Southeast Spain

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