Chronic kidney disease (CKD), secondary to renal fibrogenesis, is a public health burden. The activation of interstitial myofibroblasts and excessive production of extracellular matrix (ECM) proteins ...are major events leading to end-stage kidney disease. Recently, interleukin-15 (IL-15) has been implicated in fibrosis protection in several organs, with little evidence in the kidney. Since endogenous IL-15 expression decreased in nephrectomized human allografts evolving toward fibrosis and kidneys in the unilateral ureteral obstruction (UUO) model, we explored IL-15's renoprotective role by pharmologically delivering IL-15 coupled or not with its soluble receptor IL-15Rα. Despite the lack of effects on myofibroblast accumulation, both IL-15 treatments prevented tubulointerstitial fibrosis (TIF) in UUO as characterized by reduced collagen and fibronectin deposition. Moreover, IL-15 treatments inhibited collagen and fibronectin secretion by transforming growth factor-β (TGF-β)-treated primary myofibroblast cultures, demonstrating that the antifibrotic effect of IL-15 in UUO acts, in part, through a direct inhibition of ECM synthesis by myofibroblasts. In addition, IL-15 treatments resulted in decreased expression of monocyte chemoattractant protein 1 (MCP-1) and subsequent macrophage infiltration in UUO. Taken together, our study highlights a major role of IL-15 on myofibroblasts and macrophages, two main effector cells in renal fibrosis, demonstrating that IL-15 may represent a new therapeutic option for CKD.
Engineered bacteria are promising candidates for in situ detection and treatment of diseases. The female uro-genital tract presents several pathologies, such as sexually transmitted diseases or ...genital cancer, that could benefit from such technology. While bacteria from the gut microbiome are increasingly engineered, the use of chassis isolated from the female uro-genital resident flora has been limited. A major hurdle to implement the experimental throughput required for efficient engineering in these non-model bacteria is their low transformability. Here we report an optimized electrotransformation protocol for Lactobacillus jensenii, one the most widespread species across vaginal microflora. Starting from classical conditions, we optimized buffers, electric field parameters, cuvette type and DNA quantity to achieve an 80-fold improvement in transformation efficiency, with up to 3.5·103 CFUs/μg of DNA in L. jensenii ATCC 25258. We also identify several plasmids that are maintained and support reporter gene expression in L. jensenii. Finally, we demonstrate that our protocol provides increased transformability in three independent clinical isolates of L. jensenii. This work will facilitate the genetic engineering of L. jensenii and enable its use for addressing challenges in gynecological healthcare.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Injury to the levator ani muscle or pelvic nerves during pregnancy and vaginal delivery is responsible for pelvic floor dysfunction.
We sought to demonstrate the presence of smooth muscular cell ...areas within the levator ani muscle and describe their localization and innervation.
Five female human fetuses were studied after approval from the French Biomedicine Agency. Specimens were serially sectioned and stained by Masson trichrome and immunostained for striated and smooth muscle, as well as for somatic, adrenergic, cholinergic, and nitriergic nerve fibers. Slides were digitized for 3-dimensional reconstruction. One fetus was reserved for electron microscopy. We explored the structure and innervation of the levator ani muscle.
Smooth muscular cell beams were connected externally to the anococcygeal raphe and the levator ani muscle and with the longitudinal anal muscle sphincter. The caudalmost part of the pubovaginal muscle was found to bulge between the rectum and the vagina. This bulging was a smooth muscular interface between the levator ani muscle and the longitudinal anal muscle sphincter. The medial (visceral) part of the levator ani muscle contained smooth muscle cells, in relation to the autonomic nerve fibers of the inferior hypogastric plexus. The lateral (parietal) part of the levator ani muscle contained striated muscle cells only and was innervated by the somatic nerve fibers of levator ani and pudendal nerves. The presence of smooth muscle cells within the medial part of the levator ani muscle was confirmed under electron microscopy in 1 fetus.
We characterized the muscular structure and neural control of the levator ani muscle. The muscle consists of a medial part containing smooth muscle cells under autonomic nerve influence and a lateral part containing striated muscle cells under somatic nerve control. These findings could result in new postpartum rehabilitation techniques.
Purpose Urolithiasis after kidney transplantation can involve several contributing factors and the treatment strategy is open to question. We determined the incidence and management of urolithiasis ...in kidney recipients. Materials and Methods We retrospectively reviewed a single center series of 3,000 kidney graft recipients during 32 years to identify those with urolithiasis. We analyzed data by the prevalence per decade, including perioperative procedures (preoperative assessment, anastomosis type and urinary drainage) and long-term followup (urinary stenosis, time to presentation, size, site, treatment type, renal function and survival). Results We identified 31 cases and noted a significant decrease in incidence from 2.1% to 0.6% during the 3 decades. Excluding 4 cases of donor in situ stones the mean time to diagnosis was 8.5 years. Surgical risk factors were ureteral obstruction in 41% of cases, infravesical obstruction in 14% and urinary-digestive anastomosis in 14%. A total of 12 cases (38%) were observed exclusively with 2 of spontaneous passage. With minor adaptations all mini-invasive procedures, including extracorporeal shock wave lithotripsy, endoscopy and percutaneous nephrolithotomy, were feasible in graft recipients. Antegrade procedures were facilitated by the ventral position of the graft. Eight patients (25%) were treated with open surgical ureteroureteral anastomosis. Conclusions Prevention with a perioperative Double-J® stent and early treatment of ureteral obstruction have decreased and stabilized the urolithiasis rate at around 0.6%. Careful surveillance or any currently available instrumental treatments of urinary stones can be valid options.
Introduction
The aim of the present study was to describe autonomic urethral sphincter (US) innervation using specific muscular and neuronal antibody markers and 3D reconstruction.
Material and ...methods
We performed en‐bloc removal of the entire pelvis of three male human fetuses between 18 and 40 weeks. Serial whole mount sections (5 μm intervals) were stained and investigated. The sections were stained with Masson's trichrome and Eosin Hematoxylin, and immunostained with: anti‐SMA antibody for smooth muscle; anti‐S100 antibody for all nerves; and anti‐PMP22 antibody, anti‐TH antibody, anti‐CGRP antibody, anti‐NOS antibody for somatic, adrenergic, sensory and nitrergic nerve fibers, respectively. The slides were digitized for 3D reconstruction to improve topographical understanding. An animated reconstruction of the autonomic innervation of the US was generated.
Results
The external and internal US are innervated by autonomic nerves of the inferior hypogastric plexus (IHP). These nerves are sympathetic (positive anti‐TH antibody), sensory (positive anti‐CGRP antibody), and nitrergic (positive anti‐NOS antibody). Some autonomic fibers run within the neurovascular bundles, posterolaterally. Others run from the IHP to the posteromedial aspect of the prostate apex, above an through the rectourethral muscle. The external US is also innervated by somatic nerves (positive anti‐PMP22 antibody) arising from the pudendal nerve, joining the midline but remaining below the rectourethral.
Conclusions
This study provides anatomical evidence of an autonomic component in the innervation of the external US that travels in the neurovascular bundle. During radical prostatectomy, the rectourethral muscle and the neurovascular bundles are to be preserved, particularly during apical dissection.
Objective To evaluate the prognostic value of programmed death ligand‐1 (PD‐L1) and programmed death‐1 (PD‐1) expression in patients with upper tract urothelial carcinoma (UTUC). Patients and methods ...A retrospective multicentre study was conducted in 283 patients with UTUC treated with radical nephroureterectomy (RNU) between 2000 and 2015 at 10 French hospitals. Immunohistochemistry analyses were performed using 2 mm‐core tissue microarrays with NAT105® and 28.8® antibodies at a 5% cut‐off for positivity on tumour cells and tumour‐infiltrating lymphocytes to evaluate PD‐L1 and PD‐1 expression, respectively. Multivariable Cox regression models were used to determine the independent predictors of recurrence‐free (RFS), cancer‐specific (CSS) and overall survival (OS). Results Overall, 63 (22.3%) and 220 (77.7%) patients with UTUC had PD‐L1‐positive and ‐negative disease, respectively, while 91 (32.2%) and 192 (67.8%) had PD‐1‐positive and ‐negative disease, respectively. Patients who expressed PD‐L1 or PD‐1 were more likely to have pathological tumour stage ≥pT2 (68.3% vs 49.5%, P = 0.009; and 69.2% vs 46.4%, P < 0.001, respectively) and high‐grade (90.5% vs 70.0%, P = 0.001; and 91.2% vs 66.7%, P < 0.001, respectively) disease with lymphovascular invasion (52.4% vs 17.3%, P < 0.001; and 39.6% vs 18.2%, P < 0.001, respectively) as compared to those who did not. In multivariable Cox regression analysis adjusting for each other, PD‐L1 and PD‐1 expression were significantly associated with decreased RFS (hazard ratio HR 1.83, 95% confidence interval CI 1.09–3.08, P = 0.023; and HR 1.59, 95% CI 1.01–2.54, P = 0.049; respectively), CSS (HR 2.73, 95% CI 1.48–5.04, P = 0.001; and HR 1.96, 95% CI 1.12–3.45, P = 0.019; respectively) and OS (HR 2.08, 95% CI 1.23–3.53, P = 0.006; and HR 1.71, 95% CI 1.05–2.78, P = 0.031; respectively). In addition, multivariable Cox regression analyses evaluating the four‐tier combination of PD‐L1 and PD‐1 expression showed that only PD‐L1/PD‐1‐positive patients ( n = 38 13.4%) had significantly decreased RFS (HR 3.07, 95% CI 1.70–5.52; P < 0.001), CSS (HR 5.23, 95% CI 2.62–10.43; P < 0.001) and OS (HR 3.82, 95% CI 2.13–6.85; P < 0.001) as compared to those with PD‐L1/PD‐1‐negative disease ( n = 167 59.0%). Conclusions We observed that PD‐L1 and PD‐1 expression were both associated with adverse pathological features that translated into an independent and cumulative adverse prognostic value in UTUC patients treated with RNU.
(1)
: Understanding uterine innervation, an essential aspect of female reproductive biology, has often been overlooked. Nevertheless, the complex architecture of uterine innervation plays a ...significant role in conditions such as endometriosis. Recently, advances in histological techniques have provided unprecedented details about uterine innervation, highlighting its intricate structure, distribution, and density. The intricate nature of uterine innervation and its influence on pathologies such as endometriosis has garnered increasing attention. (2)
: This review aims to compile, analyze, and summarize the existing research on uterine innervation, and investigate its implications for the pathogenesis of endometriosis and associated pain. (3)
: A systematic review was conducted in line with PRISMA guidelines. Using the PubMed database, we searched relevant keywords such as "uterine innervation", "endometriosis", and "pain association". (4)
: The initial literature search yielded a total of 3300 potential studies. Of these, 45 studies met our inclusion criteria and were included in the final review. The analyzed studies consistently demonstrated that the majority of studies focused on macroscopic dissection of uterine innervation for surgical purposes. Fewer studies focused on micro-innervation for uterine innervation. For endometriosis, few studies focused on neural pain pathways whereas many studies underlined an increase in nerve fiber density within ectopic endometrial tissue. This heightened innervation is suggested as a key contributor to the chronic pain experienced by endometriosis patients. (5)
: The understanding of uterine innervation, and its alterations in endometriosis, offer promising avenues for research and potential treatment.
Purpose
To confront nerve dissection, tissue staining, nerve immunolabelling and Computer-Assisted Anatomic Dissection (CAAD) in identifying the precise location and origin of intrapelvic autonomic ...nerve fibers and to provide a three-dimensional (3D) representation of their relationship to other anatomical structures.
Methods
Serial transverse sections of the pelvic portion of five human female fetuses (18–31 weeks of gestation) were studied histologically (with hematoxylin/eosin and Masson trichrome) and immunohistochemically (anti-protein S100 antibody) digitized and reconstructed three-dimensionally with Surf driver software for Windows (Winsurf 4.3). Three fresh female adult cadavers were macroscopically dissected to individualize the inferior hypogastric plexus afferences and efferences and their anatomical relationships.
Results
This combined investigation including the CAAD technique allowed identifying the precise location and distribution of the pelvic nerve elements and their relationships to female pelvic organs. Hypogastric nerves (HN) were located in the retrorectal multilaminar structure and joined the homolateral inferior hypogastric plexus (IHP) at the lateral border of the recto-uterine pouch. The intersection of the ureter with the posterior wall of the uterine artery precisely located the junction of HN and IHP. Antero-inferior branches supplying female sexual and continence organs originated from the antero-inferior angle of IHP and were bundled at the posterolateral vaginal wall.
Conclusions
CAAD is an encouraging anatomical method for the development of anatomical and surgical research and teaching. Complementary to traditional anatomical studies, it may provide useful anatomical data for the comprehension of postoperative sexual and urinary dysfunction and the development of nerve-sparing surgical techniques.
Abstract Objectives To analyze to what extent partial nephrectomy (PN) is superior to radical nephrectomy (RN) in preserving renal function outcome in relation to tumor size indication. Methods and ...materials Clinical data from 973 patients operated at 9 academic institutions were retrospectively analyzed. Glomerular filtration rate (GFR) before and after surgery was calculated with the abbreviated Modification of the Diet in Renal Disease equation. For a fair comparison between the 2 techniques, all imperative indications for PN were excluded. A shift to a less favorable GFR group following surgery was considered clinically significant. Results Median age at diagnosis was 60 years (19–91). Tumor size was smaller than 4 cm in 665 (68.3%) cases and larger than 4 cm in 308 (31.7%) cases. PN and RN were performed in 663 (68.1%) and 310 (31.9%) patients, respectively. In univariate analysis, patients undergoing PN had a smaller risk for developing significant GFR change following surgery than those undergoing RN did. This was true for tumors≤4 cm ( P = 0.0001) and for tumors>4 cm ( P = 0.0001). In multivariate analysis, the following criteria were independent predictive factors for developing significant postoperative GFR loss: the use of RN ( P = 0.0001), preoperative GFR<60 ml/min ( P = 0.0001), tumor size≥4 cm ( P = 0.0001), and older age at diagnosis ( P = 0.0001). Conclusions The renal function benefit carried out by elective PN over RN persists even when expanding nephron-sparing surgery indications beyond the traditional 4-cm cutoff.
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