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1.
J Cell Biol ; 223(8)2024 Aug 05.
Article in English | MEDLINE | ID: mdl-39012627

ABSTRACT

Centrosomes are the main microtubule-organizing centers in animal cells. Due to the semiconservative nature of centrosome duplication, the two centrosomes differ in age. In asymmetric stem cell divisions, centrosome age can induce an asymmetry in half-spindle lengths. However, whether centrosome age affects the symmetry of the two half-spindles in tissue culture cells thought to divide symmetrically is unknown. Here, we show that in human epithelial and fibroblastic cell lines centrosome age imposes a mild spindle asymmetry that leads to asymmetric cell daughter sizes. At the mechanistic level, we show that this asymmetry depends on a cenexin-bound pool of the mitotic kinase Plk1, which favors the preferential accumulation on old centrosomes of the microtubule nucleation-organizing proteins pericentrin, γ-tubulin, and Cdk5Rap2, and microtubule regulators TPX2 and ch-TOG. Consistently, we find that old centrosomes have a higher microtubule nucleation capacity. We postulate that centrosome age breaks spindle size symmetry via microtubule nucleation even in cells thought to divide symmetrically.


Subject(s)
Cell Cycle Proteins , Centrosome , Microtubules , Protein Serine-Threonine Kinases , Spindle Apparatus , Centrosome/metabolism , Humans , Cell Cycle Proteins/metabolism , Cell Cycle Proteins/genetics , Spindle Apparatus/metabolism , Protein Serine-Threonine Kinases/metabolism , Protein Serine-Threonine Kinases/genetics , Microtubules/metabolism , Polo-Like Kinase 1 , Microtubule-Associated Proteins/metabolism , Microtubule-Associated Proteins/genetics , Epithelial Cells/metabolism , Cell Line , Cell Division , Tubulin/metabolism , Fibroblasts/metabolism , Antigens , Nerve Tissue Proteins
2.
Anal Chim Acta ; 1304: 342533, 2024 May 22.
Article in English | MEDLINE | ID: mdl-38637034

ABSTRACT

BACKGROUND: DIA (Data-Independent Acquisition) is a powerful technique in Liquid Chromatography coupled with high-resolution tandem Mass Spectrometry (LC-MS/MS) initially developed for proteomics studies and recently emerging in metabolomics and lipidomics. It provides a comprehensive and unbiased coverage of molecules with improved reproducibility and quantitative accuracy compared to Data-Dependent Acquisition (DDA). Combined with the Zeno trap and Electron-Activated Dissociation (EAD), DIA enhances data quality and structural elucidation compared to conventional fragmentation under CID. These tools were applied to study the lipidome and metabolome of the freshwater amphipod Gammarus fossarum, successfully discriminating stages and highlighting significant biological features. Despite being underused, DIA, along with the Zeno trap and EAD, holds great potential for advancing research in the omics field. RESULTS: DIA combined with the Zeno trap enhances detection reproducibility compared to conventional DDA, improving fragmentation spectra quality and putative identifications. LC coupled with Zeno-SWATH-DIA methods were used to characterize molecular changes in reproductive cycle of female gammarids. Multivariate data analysis including Principal Component Analysis and Partial Least Square Discriminant Analysis successfully identified significant features. EAD fragmentation helped to identify unknown features and to confirm their molecular structure using fragmentation spectra database annotation or machine learning. EAD database matching accurately annotated five glycerophospholipids, including the position of double bonds on fatty acid chain moieties. SIRIUS database predicted structures of unknown features based on experimental fragmentation spectra to compensate for database incompleteness. SIGNIFICANCE: Reproducible detection of features and confident identification of putative compounds are pivotal stages within analytical pipelines. The DIA approach combined with Zeno pulsing enhances detection sensitivity and targeted fragmentation with EAD in positive polarity provides orthogonal fragmentation information. In our study, Zeno-DIA and EAD thereby facilitated a comprehensive and insightful exploration of pertinent biological molecules associated with the reproductive cycle of gammarids. The developed methodology holds great promises for identifying informative biomarkers on the health status of an environmental sentinel species.


Subject(s)
Amphipoda , Lipidomics , Animals , Female , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Electrons , Molting , Reproducibility of Results , Metabolome , Machine Learning
3.
Anal Bioanal Chem ; 416(12): 2893-2911, 2024 May.
Article in English | MEDLINE | ID: mdl-38492024

ABSTRACT

The past decades have marked the rise of metabolomics and lipidomics as the -omics sciences which reflect the most phenotypes in living systems. Mass spectrometry-based approaches are acknowledged for both quantification and identification of molecular signatures, the latter relying primarily on fragmentation spectra interpretation. However, the high structural diversity of biological small molecules poses a considerable challenge in compound annotation. Feature-based molecular networking (FBMN) combined with database searches currently sets the gold standard for annotation of large datasets. Nevertheless, FBMN is usually based on collision-induced dissociation (CID) data, which may lead to unsatisfying information. The use of alternative fragmentation methods, such as electron-activated dissociation (EAD), is undergoing a re-evaluation for the annotation of small molecules, as it gives access to additional fragmentation routes. In this study, we apply the performances of data-dependent acquisition mass spectrometry (DDA-MS) under CID and EAD fragmentation along with FBMN construction, to perform extensive compound annotation in the crude extracts of the freshwater sentinel organism Gammarus fossarum. We discuss the analytical aspects of the use of the two fragmentation modes, perform a general comparison of the information delivered, and compare the CID and EAD fragmentation pathways for specific classes of compounds, including previously unstudied species. In addition, we discuss the potential use of FBMN constructed with EAD fragmentation spectra to improve lipid annotation, compared to the classic CID-based networks. Our approach has enabled higher confidence annotations and finer structure characterization of 823 features, including both metabolites and lipids detected in G. fossarum extracts.


Subject(s)
Amphipoda , Lipids , Metabolomics , Animals , Amphipoda/metabolism , Amphipoda/chemistry , Lipids/chemistry , Lipids/analysis , Metabolomics/methods , Lipidomics/methods , Mass Spectrometry/methods , Sentinel Species/metabolism , Electrons
4.
Sci Rep ; 14(1): 2384, 2024 01 29.
Article in English | MEDLINE | ID: mdl-38286808

ABSTRACT

Bile acids (BA) are key for liver regeneration and injury. This study aims at analyzing the changes in the BA pool induced by ischemia-reperfusion (IRI) and investigates the impact of hypothermic oxygenated perfusion (HOPE) on the BA pool compared to static cold storage (SCS). In a porcine model of IRI, liver grafts underwent 30 min of asystolic warm ischemia followed by 6 h of SCS (n = 6) ± 2 h of HOPE (n = 6) and 2 h of ex-situ warm reperfusion. The BA pool in bile samples was analyzed with liquid chromatography coupled with tandem mass spectrometry. We identified 16 BA and observed significant changes in response to ischemia-reperfusion, which were associated with both protective and injury mechanisms. Second, HOPE-treated liver grafts exhibited a more protective BA phenotype, characterized by a more hydrophilic BA pool compared to SCS. Key BA, such as GlycoCholic Acid, were identified and were associated with a decreased transaminase release and improved lactate clearance during reperfusion. Partial Least Square-Discriminant Analysis revealed a distinct injury profile for the HOPE group. In conclusion, the BA pool changes with liver graft IRI, and preservation with HOPE results in a protective BA phenotype compared to SCS.


Subject(s)
Bile Acids and Salts , Reperfusion Injury , Swine , Animals , Organ Preservation/methods , Perfusion/methods , Liver/physiology , Ischemia
5.
Article in English | MEDLINE | ID: mdl-37634392

ABSTRACT

The field of metabolomics based on mass spectrometry has grown considerably in recent years due to the need to detect and, above all, quantify a very large number of metabolites, simultaneously. Up to now, targeted multiplexed analysis on complex samples by Liquid Chromatography coupled with tandem Mass Spectrometry (LC-MS/MS) has relied almost exclusively on compound detection based on absolute retention times, as in the Scheduled-MRM (sMRM) approach. Those methods turn out to be poorly transferable from one instrument to another and result in a time-consuming and tedious method development involving a significant number of critical parameters that need specific re-optimisation. To address this challenge, we introduce a novel acquisition mode called scout-triggered MRM (stMRM). In stMRM, a marker transition is used to trigger MS analysis for a group of dependent target analytes. These marker transitions are strategically distributed throughout the chromatographic run, and the dependent analytes are associated based on their retention times. The result is a targeted assay that remains robust even in the presence of retention time shifts. A 3 to 5-fold increase in the number of detected transitions associated to plasma metabolites was obtained when transferring from a direct application of a published sMRM to a stMRM method. This significant improvement highlights the universal applicability of the stMRM method, as it can be implemented on any LC system without the need for extensive method development. We subsequently illustrate the robustness of stMRM in modified chromatographic elution conditions. Despite a large change in metabolite's selectivity, the multiplexed assay successfully recovered 70% of the monitored transitions when consequently modifying the gradient method. These findings demonstrate the versatility and adaptability of stMRM, opening new avenues for the development of highly multiplexed LC-MS/MS methods in metabolomics. These methods are characterized by their analytical transparency and straightforward implementation using existing literature data.


Subject(s)
Metabolomics , Tandem Mass Spectrometry , Chromatography, Liquid , Biological Assay , Plasma
6.
Talanta ; 253: 123806, 2023 Feb 01.
Article in English | MEDLINE | ID: mdl-36113334

ABSTRACT

Omics study exemplified by proteomics, lipidomics or metabolomics, provides the opportunity to get insight of the molecular modifications occurring in living organisms in response to contaminants or in different physiological conditions. However, individual omics discloses only a single layer of information leading to a partial image of the biological complexity. Multiplication of samples preparation and processing can generate analytical variations resulting from several extractions and instrumental runs. To get all the -omics information at the proteins, metabolites and lipids level coming from a unique sample, a specific sample preparation must be optimized. In this study, we streamlined a biphasic extraction procedure based on a MTBE/Methanol mixture to provide the simultaneous extraction of polar (proteins, metabolites) and apolar compounds (lipids) for multi-omics analyses from a unique biological sample by a liquid chromatography (LC)/mass spectrometry (MS)/MS-based targeted approach. We applied the methodology for the study of female amphipod Gammarus fossarum during the reproductive cycle. Multivariate data analyses including Partial Least Squares Discriminant Analysis and multiple factor analysis were applied for the integration of the multi-omics data sets and highlighted molecular signatures, specific to the different stages.


Subject(s)
Multiomics , Proteomics , Female , Humans , Data Analysis , Lipidomics , Metabolomics
7.
STAR Protoc ; 3(3): 101493, 2022 09 16.
Article in English | MEDLINE | ID: mdl-35776653

ABSTRACT

Drosophila neural stem cells (NSCs) divide asymmetrically to generate siblings of different sizes. This model system has proved helpful in deciphering the contribution of polarity cues and the mitotic spindle in asymmetric cell division. Here, we describe a technique we developed to flatten cultured Drosophila brain explants to accurately image the cytoskeleton in live NCSs. We also describe our approach to efficiently remove centrosomes by laser photo-ablation and to measure daughter cell size after NSC division. For complete details on the use and execution of this protocol, please refer to Thomas et al. (2021).


Subject(s)
Drosophila Proteins , Neural Stem Cells , Animals , Centrosome , Drosophila , Drosophila melanogaster
8.
Cell Rep ; 37(4): 109895, 2021 10 26.
Article in English | MEDLINE | ID: mdl-34706235

ABSTRACT

Neuroblast division is characterized by asymmetric positioning of the cleavage furrow, resulting in a large difference in size between the future daughter cells. In animal cells, furrow placement and assembly are governed by centralspindlin that accumulates at the equatorial cell cortex of the future cleavage site and at the spindle midzone. In neuroblasts, these two centralspindlin populations are spatially and temporally separated. A leading pool is located at the basal cleavage site and a second pool accumulates at the midzone before traveling to the cleavage site. The cortical centralspindlin population requires peripheral astral microtubules and the chromosome passenger complex for efficient recruitment. Loss of this pool does not prevent cytokinesis but enhances centralspindlin signaling at the midzone, leading to equatorial furrow repositioning and decreased size asymmetry. These data show that basal furrow positioning in neuroblasts results from a competition between different centralspindlin pools in which the cortical pool is dominant.


Subject(s)
Cytokinesis , Microtubules , Neural Stem Cells , Animals , Drosophila melanogaster , Microtubules/genetics , Microtubules/metabolism , Microtubules/ultrastructure , Neural Stem Cells/metabolism , Neural Stem Cells/ultrastructure
9.
Curr Biol ; 31(4): 684-695.e6, 2021 02 22.
Article in English | MEDLINE | ID: mdl-33259793

ABSTRACT

Proper assembly of mitotic spindles requires microtubule nucleation not only at the centrosomes but also around chromatin. In this study, we found that the Drosophila tubulin-specific chaperone dTBCE is required for the enrichment of tubulin in the nuclear space after nuclear envelope breakdown and for subsequent promotion of spindle microtubule nucleation. These events depend on the CAP-Gly motif found in dTBCE and are regulated by Ran and lamin proteins. Our data suggest that during early mitosis, dTBCE and nuclear pore proteins become enriched in the nucleus, where they interact with the Ran GTPase to promote dynamic tubulin enrichment. We propose that this novel mechanism enhances microtubule nucleation around chromatin, thereby facilitating mitotic spindle assembly.


Subject(s)
Chromatin , Microtubules , Tubulin , Animals , Drosophila , Mitosis , Spindle Apparatus , Tubulin/genetics , Tubulin/metabolism
10.
J Cell Sci ; 133(7)2020 04 06.
Article in English | MEDLINE | ID: mdl-32094264

ABSTRACT

A novel 2,3-benzodiazepine-4 derivative, named 1g, has recently been shown to function as an anti-proliferative compound. We now show that it perturbs the formation of a functional mitotic spindle, inducing a spindle assembly checkpoint (SAC)-dependent arrest in human cells. Live analysis of individual microtubules indicates that 1g promotes a rapid and reversible reduction in microtubule growth. Unlike most anti-mitotic compounds, we found that 1g does not interfere directly with tubulin or perturb microtubule assembly in vitro The observation that 1g also triggers a SAC-dependent mitotic delay associated with chromosome segregation in Drosophila neural stem cells, suggests that it targets a conserved microtubule regulation module in humans and flies. Altogether, our results indicate that 1g is a novel promising anti-mitotic drug with the unique properties of altering microtubule growth and mitotic spindle organization.


Subject(s)
Benzodiazepines , Mitosis , Benzodiazepines/pharmacology , Humans , Microtubules , Spindle Apparatus , Tubulin/genetics
11.
Nephrol Ther ; 14(3): 148-152, 2018 May.
Article in French | MEDLINE | ID: mdl-29198962

ABSTRACT

We report the cases of three young patients suffering from type 1 primary hyperoxaluria, a metabolic genetic disorder characterized by intracellular accumulation of oxalate and which may result in end-stage renal disease with systemic impairment. A number of effective conservative therapeutic means are available for early management of affected children particularly when he is growing older. Despite the demonstrated efficacy of conservative therapy, compliance represents a major and daily challenge. Monitoring therapeutic compliance is thus an important task for physicians in charge of this disease. A better understanding of non-compliance causes is required to improve the follow-up of patients for whom treatment education must be a priority.


Subject(s)
Conservative Treatment/methods , Hyperoxaluria, Primary/therapy , Medication Adherence , Adult , Child , Child, Preschool , Female , Humans , Hyperoxaluria, Primary/diagnosis , Male , Mutation , Transaminases/genetics , Urinalysis/methods
12.
J Endourol ; 31(5): 476-481, 2017 May.
Article in English | MEDLINE | ID: mdl-28292198

ABSTRACT

Purpose: To propose and evaluate a new endoscopic technique using only a retrograde ureteroscopic approach for the removal of heavily encrusted ureteral stents. Methods: Data from 51 consecutive patients with encrusted and retained ureteral stents were prospectively collected. Description of the successive steps of surgery is detailed. The holmium yttrium aluminum garnet (Ho:YAG) laser properties offer the opportunity for fragmentation of stent-attached encrustation and the ability to cut the stent itself. Reducing the length of the stent is critical to creating space in the ureter and to allow free access for ureteroscopes or ureteral access sheath placement. The primary outcome of this study was the feasibility and the safety of this retrograde intrarenal approach. Some factors of encrustation and outcomes are also discussed in comparison with lithotripsy, percutaneous, laparoscopic, open surgery, or a combination of these techniques. Results: The removal of the encrusted stent was possible with only this retrograde technique in 98% of patients. The transection of the encrusted stent with the Ho:YAG laser was useful in 71% of the patients. Mean operative time was 110 minutes and mean hospital stay was 2.33 days. Postoperative complications were mainly nonobstructive pyelonephritis (10%). The most significant predictor of this life-threatening complication was the presence of struvite stones with the encrusted stent (p = 0.018). Contrariwise, operative time, body mass index, gender, and encrustation rate were not associated with postoperative pyelonephritis. Both cystine stone disease and pregnancy led to faster stent encrustation. Conclusions: Retrograde ureteroscopic surgery is efficient and safe for removing retained stents and associated stone burdens. The Ho:YAG laser is essential to perform the encrustation removal and sectioning of the stent.

13.
Urology ; 102: 38-42, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27765587

ABSTRACT

OBJECTIVE: To assess the outcomes of patients following a first-line systematic endourologic procedure used to treat ureteroenteric anastomotic strictures (UEAS). MATERIALS AND METHODS: All data from patients treated using a first-line endourologic approach for UEAS between 2010 and 2015 were reviewed retrospectively. The following data were analyzed: age, type of urinary diversion, initial symptoms, surgical endoscopic approach (antegrade or retrograde), pre- and postoperative creatinine levels, and postoperative complications and outcomes. Follow-up visits occurred at 6 weeks, 3 months, and 6 months postoperatively, and at least annually thereafter. RESULTS: A total of 27 patients (median age: 62.5 years) were included. Overall, 28 UEAS were treated endoscopically (ileal conduit: n = 25; neobladder: n = 3). Most UEAS developed following radical cystectomy for bladder cancer (n = 19). Overall, the endoscopic approach was successful in 20 cases (71.4%). The UEAS length was >1 cm in 21 cases (75%). All UEAS of <1 cm were treated successfully (n = 7). There were three grade II and five grade III complications. The median follow-up period was 25 months. The median creatinine levels before surgery and at last follow-up were 1.3 mg/dL and 0.9 mg/dL, respectively. CONCLUSION: An endourologic procedure is a reasonable option for first-line treatment for UEAS and has promising functional outcomes and limited morbidity.


Subject(s)
Ileum/surgery , Postoperative Complications/surgery , Ureter/surgery , Urinary Diversion , Adolescent , Adult , Anastomosis, Surgical , Constriction, Pathologic/surgery , Female , Humans , Male , Retrospective Studies , Treatment Outcome , Ureteroscopy , Young Adult
14.
Nephrol Ther ; 11(1): 42-9, 2015 Feb.
Article in French | MEDLINE | ID: mdl-25488796

ABSTRACT

Urolithiasis is a common condition, with a prevalence of ∼10% and a male/female ratio above 1 according to large national series. Various types of urinary stones have been described upon their mineral content and/or their morphology. Hence, a combined morpho-constitutional (M-C) classification has been proposed. In order to detail the prevalence of urolithiasis in general and of each M-C type in particular upon age and gender in Belgium, we retrospectively studied the database of a reference center for urolithiasis analysis. Between 2010 and 2013, 2195 stones were characterized. We excluded 45 non-biological stones and 281 stones, which originated from outside the study zone. Among 1869 stones, 1293 (69.2%) affected men. Prevalence peak of urolithiasis was observed between 50-60 years of age in both genders. The M-C analysis was available for 1854 stones (99.2%): multiple morphological types were concomitantly identified in 49.3%. In the whole population, the main mineral constituent was whewellite (54.4%), mainly organized as type Ia (94%). Weddellite was found in 19.8%, with an equal distribution between types IIa and IIb. Uric acid was the 3rd most frequent constituent in man, with a similar distribution between IIIa and IIIb. Phosphate was uncommon in man (8.2%), but frequent in woman (26.6%) with a type IVa1 organization. Prevalence of M-C types changes with aging, i.e. decrease of weddellite and increase of whewellite and uric acid in both genders. This retrospective analysis of a single-center database of urinary stones helps characterize the M-C epidemiology of urolithiasis in Belgium.


Subject(s)
Urolithiasis/classification , Urolithiasis/epidemiology , Belgium/epidemiology , Female , Humans , Male , Middle Aged , Retrospective Studies
15.
J Endourol ; 28(3): 286-90, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24147776

ABSTRACT

PURPOSE: Our aim is to evaluate different ureteral access sheaths (UASs), which are available in the international market and their compatibility with different available flexible ureteroscopes (F-URSs) to help the urologist choose the proper ureteral access sheath for his or her endoscope before commencing the procedure. MATERIALS AND METHODS: A total of 21 UASs and 12 F-URSs were evaluated. Measurements were obtained in French (F) units considering different characteristics for each UAS and each F-URS. Insertion test without friction between F-URS and UAS was considered as a successful test and was referred as (YES). RESULTS: All UASs and F-URSs were successfully submitted to the insertion test. All F-URSs that were inserted into UASs without friction had an internal diameter of at least 12F. Different lengths of UAS did not influence the test outcome. CONCLUSION: This study was able to establish a correlation table between different UASs and different flexible ureteroscopes. As of now, the 12/14F UAS is considered the universal UAS that accepts all F-URSs that are available in the endourology field. Nevertheless, we are expecting a significant change with the new standard size 10/12F UAS as well as huge advances in minimizing the size of different endoscopes.


Subject(s)
Kidney Calculi/surgery , Ureteroscopes , Ureteroscopy/methods , Equipment Design , Humans , Reproducibility of Results
16.
J Urol ; 189(2): 580-4, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22982421

ABSTRACT

PURPOSE: The safety of using a ureteral access sheath during retrograde intrarenal surgery remains controversial. Using a novel classification, we prospectively evaluated the incidence and severity of ureteral access sheath driven ureteral wall injury after flexible ureteroscopy for retrograde intrarenal surgery. MATERIALS AND METHODS: Data on a total of 359 consecutive patients who underwent retrograde intrarenal surgery for kidney stone were prospectively collected at 2 academic centers. We propose what is to our knowledge a novel endoscopic classification of iatrogenic ureteral wall injury. Ureteral injuries after retrograde intrarenal surgery were assessed visually with a digital flexible ureterorenoscope. The primary outcome measure was the incidence and nature of ureteral injuries. We sought factors predisposing to such injuries. RESULTS: Ureteral wall injury was found in 167 patients (46.5%). Severe injury involving the smooth muscle layers was observed in 48 patients (13.3%). Males vs females (p = 0.024) and older vs younger patients (p = 0.018) were at higher risk for severe ureteral access sheath related ureteral injury. The most significant predictor of severe injury was absent ureteral Double-J® stenting before retrograde intrarenal surgery (p <0.0001). Pre-stenting vs no pre-stenting decreased the risk of severe injury by sevenfold. Body mass index, a history of diabetes mellitus, vascular disease or abdominopelvic radiation therapy and operative time were not associated with severe ureteral injury. CONCLUSIONS: Ureteral access sheath use for retrograde intrarenal surgery should involve systematic visual assessment of the entire ureter to recognize severe ureteral injury. The incidence of severe ureteral injury is largely decreased by preoperative Double-J stenting.


Subject(s)
Intraoperative Complications/classification , Intraoperative Complications/etiology , Stents , Ureter/injuries , Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Incidence , Injury Severity Score , Intraoperative Complications/epidemiology , Male , Middle Aged , Prospective Studies , Urologic Surgical Procedures/instrumentation , Young Adult
17.
Int Urogynecol J ; 22(2): 145-56, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20857087

ABSTRACT

INTRODUCTION AND HYPOTHESIS: The aim of this study is to compare a modified inside-out transobturator procedure with its original counterpart [inside-out transobturator (TVT-O)] for the treatment of female stress urinary incontinence (SUI). METHODS: A prospective, randomized trial in women suffering from SUI was used. The modified procedure consisted of a shorter tape whilst the scissors or guide no longer perforated the obturator membrane. The primary outcome was the resolution of subjective and objective SUI at 1 year. Secondary outcome measures included adverse events, quality of life measures, and groin pain. RESULTS: One hundred seventy-five patients were randomized. No intraoperative complications were recorded. The SUI cure rate was 91.7% versus 90.7% (original versus modified, respectively; p = 0.824). Incidence and intensity of groin pain was higher in the original TVT-O group on day 0 and 1 (p < 0.05), requiring more analgesics (p = 0.015) but not thereafter. CONCLUSIONS: At 1 year follow-up, the modified inside-out transobturator tape procedure was as efficient and safe as the original technique but associated with less immediate postoperative groin pain.


Subject(s)
Gynecologic Surgical Procedures/methods , Suburethral Slings , Urinary Incontinence, Stress/surgery , Adult , Aged , Aged, 80 and over , Belgium/epidemiology , Female , Gynecologic Surgical Procedures/adverse effects , Humans , Middle Aged , Pain, Postoperative/epidemiology , Pain, Postoperative/etiology , Prospective Studies , Treatment Outcome
18.
Eur Urol ; 58(1): 96-104, 2010 Jul.
Article in English | MEDLINE | ID: mdl-19674832

ABSTRACT

BACKGROUND: Accumulating evidence suggests that DNA methylation markers could serve as sensitive and specific cancer biomarkers. OBJECTIVE: To determine whether a panel of methylated genes would have the potential to identify primary bladder cancer (BCa) in voided urine samples. DESIGN, SETTING, AND PARTICIPANTS: A pharmacologic unmasking reexpression analysis in BCa cell lines was initially undertaken to unveil candidate methylated genes, which were then evaluated in methylation-specific polymerase chain reaction (MSP) assays performed on DNA extracted from noncancerous and cancerous bladder tissues. The most frequently methylated genes in cancerous tissues, with 100% specificity, were retained for subsequent MSP analysis in DNA extracted from urine samples to build and validate a panel of potential methylated gene markers. Urine samples were prospectively collected at three urologic centres from patients with histologically proven BCa and processed for use in real-time MSP and cytologic analysis. Patients with nonmalignant urologic disorders were included as controls. MEASUREMENTS: A urine sample was classified as valid when > or = 10 copies of the gene encoding ß-actin were measured in the urine sediment genomic DNA. Sensitivity, specificity, and predictive values of the MSP and cytology tests were assessed and compared. RESULTS AND LIMITATIONS: MSP assays performed on 466 of the 496 (94%) valid urine samples identified two genes, TWIST1 and NID2, that were frequently methylated in urine samples collected from BCa patients, including those with early-stage and low-grade disease. The sensitivity of this two-gene panel (90%) was significantly better than that of cytology (48%), with comparable specificity (93% and 96%, respectively). The positive predictive value and negative predictive value of the two-gene panel was 86% and 95%, respectively. CONCLUSIONS: Detection of the methylated TWIST1 and NID2 genes in urine sediments using MSP provides a highly (> or = 90%) sensitive and specific, noninvasive approach for detecting primary BCa. TRIAL REGISTRATION: BlCa-001 study - EudraCt 2006-003303-40.


Subject(s)
Biomarkers, Tumor/genetics , Cell Adhesion Molecules/genetics , DNA Methylation , DNA, Neoplasm/urine , Nuclear Proteins/genetics , Twist-Related Protein 1/genetics , Urinary Bladder Neoplasms/diagnosis , Actins/genetics , Actins/urine , Adult , Aged , Aged, 80 and over , Calcium-Binding Proteins , Cell Line, Tumor , Female , Humans , Male , Middle Aged , Neoplasm Staging , Polymerase Chain Reaction , Prospective Studies , Sensitivity and Specificity , Urinary Bladder Neoplasms/urine
19.
Rev Bras Ortop ; 45(6): 577-82, 2010.
Article in English | MEDLINE | ID: mdl-27026967

ABSTRACT

OBJECTIVE: To evaluate the clinical and functional outcomes from patients undergoing surgery to treat hip fractures, with regard to the ASA score and time spent waiting for definitive surgical treatment. METHOD: Over a one-year period, 154 patients with hip fractures, aged 65 years and over, underwent operations. Data on the preoperative ASA score and the time spent waiting for the operation were obtained. Two years after the operation, Zuckerman's Functional Recovery Score (FRS) questionnaire was used to assess the patients' current functional capacity. RESULTS: Mortality during the first postoperative year differed between patients with ASA 3 or 4 and those classified as ASA 1 or 2 (significant data; p < 0.05). Mortality up to the end of the second postoperative year was significantly higher (p < 0.05) in the ASA 3 or 4 group. The preoperative ASA score did not demonstrate any significant relationship with the patients' current functional capacity (p > 0.05). There was no significant difference between the group operated within 48 hours of admission and the group operated after 48 hours, in relation to mortality or current functional capacity (p > 0.05). The group aged 80 years and over showed significantly higher mortality (p < 0.05) than the group aged 65 to 79 years up to the end of the second postoperative year. CONCLUSION: The preoperative ASA score and an age of 80 years or over may be considered to be factors associated with higher mortality two years after hip fracture surgery. In isolation, time spent waiting for surgery was not significant.

20.
Rev. bras. ortop ; 45(6): 577-582, 2010. tab
Article in Portuguese | LILACS | ID: lil-574823

ABSTRACT

OBJETIVO: Avaliar o resultado clínico e funcional dos pacientes submetidos à cirurgia por fratura do quadril relacionando com o escore ASA e com tempo para o tratamento cirúrgico definitivo. MÉTODOS: No período de um ano foram operados 154 pacientes com 65 anos ou mais, com fratura do quadril. Dados sobre o escore pré-operatório ASA e sobre o tempo de espera para a cirurgia foram obtidos. Dois anos após a cirurgia, foi aplicado o questionário Escore de Recuperação Funcional de Zuckerman (ERF) como examinadorda capacidade funcional atual dos pacientes. RESULTADOS: A mortalidade no primeiro ano pós-operatório apresentou diferença entre pacientes com ASA 3 ou 4 em relação aos classificados como ASA 1 ou 2 (dado significante p < 0,05). A mortalidade até o final do segundo ano pós-operatório foi significantemente maior (p < 0,05) no grupo ASA 3 ou 4. O escore pré-operatório ASA não apresentou relação significante com a capacidade funcional atual dos pacientes (dado significante p < 0,05). Não houve diferença significante entre o grupo operado em menos de 48 horas da admissão do grupo operado após 48 horas, em relação à mortalidade e à capacidade funcional atual (dado significante p < 0,05). O grupo de 80 anos ou mais apresentou mortalidade significantemente maior (p < 0,05) do que o grupo de 65 a 79 anos até o final do segundo ano pós-operatório. CONCLUSÃO: O escore pré-operatório ASA e a idade maior de 80 anos podem ser considerados fatores associados à maior mortalidade após dois anos de pós-operatório por fratura do quadril. Isoladamente, o tempo para o tratamento cirúrgico não foi significante.


OBJECTIVE: To evaluate the clinical and functional outcomes of patients submitted to surgery for hip fracture through the ASA score and time for definitive surgical treatment. METHOD: During one year, 154 patients with hip fractures, aged 65 years and over, were operated on. Data from the pre-operative ASA score and time awaiting operation were obtained. Two years after the operation, Zuckerman's Functional Recovery Score (FRS) questionnaire was used to assess the current functional capacity of the patients. RESULTS: Mortality in the first post-operative year was different between patients with an ASA 3 or 4 compared to those classified as ASA 1 or 2 group (significant data p<0.05). Mortality up to the end of the second post-operative year was significantly higher (p<0.05) in the ASA 3 or 4 group. The preoperative ASA score didn't demonstrate a significant relationship to the current functional capacity of the patients (p>0.05). There was no significant difference between the group operated within 48 hours of admission and the group operated after 48 hours in relation to mortality or current functional capacity (p>0.05). The group aged 80 and over showed significantly higher mortality than the group aged 65 to 79 years through the end of the second post-operative year (p>0.05). CONCLUSION: A preoperative ASA score and an age of 80 or over may be considered factors associated with higher mortality two years post-operatively after hip fracture. In isolation, time awaiting surgery was not significant.


Subject(s)
Humans , Male , Female , Aged , Aged, 80 and over , Hip Fractures/surgery , Hip Fractures/rehabilitation , Health of the Elderly , Postoperative Period
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