From fix to fit into the autoptic human brains.

Formalin-fixed, paraffinembedded (FFPE) human brain tissues are very often stored in formalin for long time. Formalin fixation reduces immunostaining, and the DNA/RNA extraction from FFPE brain tissue becomes suboptimal. At present, there are different protocols of fixation and several procedures and kits to extract DNA/RNA from paraffin embedding tissue, but a gold standard protocol remains distant. In this study, we analyzed four types of fixation systems and compared histo and immuno-staining. Based on our results, we propose a modified method of combined fixation in formalin and formic acid for the autoptic adult brain to obtain easy, fast, safe and efficient immunolabelling of long-stored FFPE tissue. In particular, we have achieved an improved preservation of cellular morphology and obtained success in postmortem immunostaining for NeuN. This nuclear antigen is an important marker for mapping neurons, for example, to evaluate the histopathology of temporal lobe epilepsy or to draw the topography of cardiorespiratory brainstem nuclei in sudden infant death syndrome (SIDS). However, NeuN staining is frequently faint or lost in postmortem human brain tissues. In addition, we attained Fluoro Jade C staining, a marker of neurodegeneration, and immunofluorescent staining for stem cell antigens in the postnatal human brain, utilizing custom fit fixation procedures

Continuous stellate ganglion block in delayed cerebral ischemia: A possible supplementary approach to traditional therapy?

Delayed Cerebral Ischemia (DCI) is a major contributor to morbidity and mortality after SAH. Currently the prevention of vasospasm and DCI relies on nimodipine administration and on maintaining an adequate cerebral perfusion pressure. We report a patient with initial DCI after SAH in which stellate ganglion block (SGB) was performed after nimodipine administration. Firstly the procedure was characterized by a iv and intra-arterial nimodipine administration which did not result into a normal perfusion pattern. Therefore a single-shot stellate ganglion block was performed, as suggested in literature. Because of the not sufficient but promising perfusion improvement, we decided to deliver a continuous ganglion block (cSGB) for 5 days. Consequently a further improvement of the cerebral perfusion on CTPerfusion and Real Time Angiographic Perfusion Assessment was registered. In order to treat cerebral vasospasm, SGB is known to be a further valuable treatment, despite its temporary effect. However the continuous use of SGB during initial DCI has never been described before

Physical Study by Surface Characterizations of Sarin Sensor on the Basis of Chemically Functionalized Silicon Nanoribbon Field Effect Transistor

Surface characterizations of an organophosphorus (OP) gas detector based on chemically functionalized silicon nanoribbon field-effect transistor (SiNR-FET) were performed by Kelvin Probe Force Microscopy (KPFM) and ToF-SIMS, and correlated with changes in the current-voltage characteristics of the devices. KPFM measurements on FETs allow (i) to investigate the contact potential difference (CPD) distribution of the polarized device as function of the gate voltage and the exposure to OP traces and, (ii) to analyze the CPD hysteresis associated to the presence of mobile ions on the surface. The CPD measured by KPFM on the silicon nanoribbon was corrected due to side capacitance effects in order to determine the real quantitative surface potential. Comparison with macroscopic Kelvin probe (KP) experiments on larger surfaces was carried out. These two approaches were quantitatively consistent. An important increase of the CPD values (between + 399 mV and + 302 mV) was observed after the OP sensor grafting, corresponding to a decrease of the work function, and a weaker variation after exposure to OP (between - 14 mV and - 61 mV) was measured. Molecular imaging by ToF-SIMS revealed OP presence after SiNR-FET exposure. The OP molecules were essentially localized on the Si-NR confirming effectiveness and selectivity of the OP sensor. A prototype was exposed to Sarin vapors and succeeded in the detection of low vapor concentrations (40 ppm).Comment: Paper and supporting information, J. Phys. Chem. C, 201

Role of d-mannose in the prevention of recurrent uncomplicated cystitis: State of the art and future perspectives

Background: Urinary tract infections (UTI) are highly frequent in women, with a significant impact on healthcare resources. Although antibiotics still represent the standard treatment to manage recurrent UTI (rUTI), D-mannose, an inert monosaccharide that is metabolized and excreted in urine and acts by inhibiting bacterial adhesion to the urothelium, represents a promising nonantibiotic prevention strategy. The aim of this narrative review is to critically analyze clinical studies reporting data concerning the efficacy and safety of D-mannose in the management of rUTIs. Methods: A nonsystematic literature search, using the Pubmed, EMBASE, Scopus, Web of science, Cochrane Central Register of Controlled Trials, and Cochrane Central Database of Systematic Reviews databases, was performed for relevant articles published between January 2010 and January 2021. The following Medical Subjects Heading were used: “female/woman”, “urinary tract infection”, and “D-mannose”. Only clinical studies, systematic reviews, and meta-analyses reporting efficacy or safety data on D-mannose versus placebo or other competitors were selected for the present review. Evidence was limited to human data. The selected studies were organized in two categories according to the presence or not of a competitor to D-mannose. Results: After exclusion of non-pertinent studies/articles, 13 studies were analyzed. In detail, six were randomized controlled trials (RCTs), one a randomized cross-over trial, five prospective cohort studies, and one a retrospective analysis. Seven studies compared D-mannose to placebo or others drugs/dietary supplements. Six studies evaluated the efficacy of D-mannose comparing follow-up data with the baseline. D-mannose is well tolerated, with few reported adverse events (diarrhea was reported in about 8% of patients receiving 2 g of D-mannose for at least 6 months). Most of the studies also showed D-mannose can play a role in the prevention or rUTI or urodynamics-associated UTI and can overlap antibiotic treatments in some cases. The possibility to combine D-mannose with polyphenols or Lactobacillus seems another important option for UTI prophylaxis. However, the quality of the collected studies was very low, generating, consequently, a weak grade of recommendations as suggested by international guidelines. Data on D-mannose dose, frequency, and duration of treatment are still lacking. Conclusion: Dmannose alone or in combination with several dietary supplements or Lactobacillus has a potential role in the non antimicrobial prophylaxis or recurrent UTI in women. Despite its frequent prescription in real-life practice, we believe that further well-designed studies are urgently needed to definitively support the role of D-mannose in the management of recurrent UTIs in women

Harmonization in preclinical epilepsy research: A joint AES/ILAE translational initiative

Among the priority next steps outlined during the first translational epilepsy research workshop in London, United Kingdom (2012), jointly organized by the American Epilepsy Society (AES) and the International League Against Epilepsy (ILAE), are the harmonization of research practices used in preclinical studies and the development of infrastructure that facilitates multicenter preclinical studies. The AES/ILAE Translational Task Force of the ILAE has been pursuing initiatives that advance these goals. In this supplement, we present the first reports of the working groups of the Task Force that aim to improve practices of performing rodent videoâ\u80\u93electroencephalography (vEEG) studies in experimental controls, generate systematic reviews of preclinical research data, and develop preclinical common data elements (CDEs) for epilepsy research in animals

Development and evaluation of a methodology to integrate technical and sensorial properties in materials selection.

In the materials selection process, the use of different tools, languages and perspectives frequently causes disagreement between engineers and industrial designers. The aim of the paper is to define an integrated method for materials selection that provides industrial designers with measurable data to support and explain aesthetic decisions on materials. A new method for materials selection consisting of multiple tools structured in a two-step framework is presented. The method is tested through a case study of professional kitchen appliances where metal components are replaced with polymers. The first step involved the application of an established technique to identify polymeric bulk solutions, based on their technical properties. The second step employed a sensory analysis test to choose suitable finishes. Thirty-seven individuals performed the test: the subjects highlighted their main perceptions of metal and metal-look polymer finishes. The research demonstrates that the proposed method is suitable for the evaluation of both technical and sensorial properties of materials. In particular, Mapping test represents a rapid, low cost and effective tool to help industrial designers justify Colour Materials and Finish (CMF) choices with quantifiable information

EGFR CELL EXPRESSION IN BLADDER WASHINGS AS A RISK MARKER TOOL IN NON MUSCLE-INVASIVE BLADDER CANCER. PRELIMINARY EXPERIENCE

INTRODUCTION AND OBJECTIVES: Up to day, EGFR expression has been determined mainly in tissue specimens of muscleinvasive bladder cancer and its overexpression has been associated with worse prognosis and shorter survival. Urothelial EGFR status after NMIBC transurethral resection (TUR) could indicate the risk of recurrence and progression. We investigated the feasibility of EGFR measurement in bladder washings of patients undergoing intravesical adjuvant therapy for NMIBC and its usefulness in identifying risk subgroups. METHODS: Our prospective study included patients after TUR of NMIBC and healthy controls. A cellular pellet was obtained from bladder washing, and RNA extraction performed by miRNeasy Mini Kit (Qiagen). Good quality of RNA was checked. The cDNA obtained from RNA was used to perform a gene expression analysis by a Real Time PCR, according to the method of the comparative quantification (DDCt) with an endogenous control (Cyclophilin). Every reaction was set in triplicate as a guarantee of quality. Patients were grouped for EAU risk class and maintained in follow-up. The EGFR expressions were statistically analyzed according to EAU risk groups and to patients0 outcome. EGFR gene expression values were expressed in FOLDs of change compared to healthy controls (EGFR¼1). RESULTS: Fifty-eight patients and 21 healthy age-matched controls were entered. An adequate cellular pellet was obtained in 50 patients (86.2%) showing a median EGFR expression of 2.0 folds (IQR 0.6-4.3, p¼0.0004). After TUR and adjuvant intravesical therapy, 22 (55%) out of 40 high-risk patients, showed EGFR decrease to 1.3 folds (IQR 0.9-1.5), while 18 (45%) showed elevated EGFR, median 4.7 (IQR 4.1-11.6). At 25 months median follow-up (IQR 19.0-34.8), 20 (40%) patients recurred and 6 (12%) progressed. Among patients with or without EGFR gene increase, 9 (22.5%) and 5 (12.5%) recurred and 5 (12.5%) and 1 (2.5%) progressed, respectively. CONCLUSIONS: In our experience EGFR expression measurement was feasible in more than 85% of patients and resulted related to EAU risk classes for recurrence and progression, showing different behavior during intravesical therapy. It was possible to identify a subgroup of high risk patients overexpressing EGFR in spite of intravesical adjuvant therapy. EGFR evaluation in bladder washing could represent a repeatable and useful tool to identify a subgroup of patients at risk for progression unresponsive to intravesical adjuvant therapy and candidate to early radical cystectom

The activity of intravesical hyaluronic acid and chondroitin sulfate administration on urothelial gene expression. Preliminary results on the epidermal growth factor receptor and fibronectin gene expression evaluated in bladder washings of patients affected by non muscle-invasive bladder cancer

Introduction & Objectives Hyaluronic acid (HA) and chondroitin sulfate (CS) are two major constituents of the bladder glycosaminoglycan layer. Recent data show that Fibronectin (FN) and Epidermal Growth Factor Receptor (EGFR) gene expression can be measured in bladder washings and could represent potential biomarkers of urothelial damage and tumor aggressiveness, respectively (1,2). The aim of our study was to investigate the interference of a single intravesical instillation of HA-CS solution on the expression of FN and EGFR genes in patients affected by non-muscle-invasive bladder cancer (NMIBC). Material & Methods A prospective double-blinded study included patients undergoing adjuvant intravesical therapy for NMIBC and age matched healthy controls. For EGFR evaluation, a single HA-CS solution was administered intravesically 14 days after transurethral resection of high risk NMIBC, before the start of the adjuvant therapy. For FN evaluation, a single HA-CS instillation was administered to patients showing local toxicity secondary to intravesical adjuvant therapy. Samples of bladder washings were collected before and one week after the HA-CS instillation, obtaining a cellular pellet stored at -80 °C. Cellular RNA was isolated by a miRNeasy Mini Kit (Qiagen®) and cDNA, obtained using a “High Capacity cDNA Reverse Transcription Kit” (Life Technologies®) was used to perform a gene expression analysis by a Real Time PCR. EGFR and FN gene expression values were expressed in FOLDs of change compared to healthy controls (FN and EGFR=1). Results Thirty-eight patients and 5 controls entered the study. Seventeen and 21 patients were evaluated for FN and EGFR respectively. In 21 patients with high risk NMIBC, the median EGFR expression decreased from 2.4 folds (range: 0.1-39.0) to 1.0 fold (range: 0.05-36.8) showing a statistically significant decrease of 58.3% (p<0.02). In patients showing clinically relevant toxicity secondary to intravesical adjuvant therapy (BCG in 9 and Epirubicin in 8 patients) the median FN expression value dropped from 1.8 folds (range: 0.07-8.1) to 0.9 fold (range: 0.1-7.5) after HA-CS administration with a statistically significant decrease of 50% (p<0.05). Conclusions FN gene expression in bladder washings appears related to the intensity of the urothelial damage, reaching higher expression levels in case of severe toxicity induced by intravesical adjuvant therapy (2). In our experience the FN gene expression significantly decreases a week after the administration of HA-CS solution with contemporary symptomatic relief. Moreover the urothelial EGFR gene expression resulted significantly lowered one week after the HA-CS intravesical administration. The reduced availability of its receptor could limit the proliferative activity of EGF on the urothelium promoting recurrence and progression. Acknowledgements: GSTU Foundation References: 1. Serretta V, et al. Feasibility of EGFR evaluation in bladder washings of patients affected by non muscle-invasive bladder cancer. J Urol, 2016. 195 (4S): e327. 2. Alonge V, et al. Correlation between Fibronectin gene expression and local toxicity induced by adjuvant intravesical therapy. J Urol, 2015. 193 (4S): e53

Comparison between Two Different Two-Stage Transperineal Approaches to Treat Urethral Strictures or Bladder Neck Contracture Associated with Severe Urinary Incontinence that Occurred after Pelvic Surgery: Report of Our Experience

Introduction. The recurrence of urethral/bladder neck stricture after multiple endoscopic procedures is a rare complication that can follow prostatic surgery and its treatment is still controversial. Material and Methods. We retrospectively analyzed our data on 17 patients, operated between September 2001 and January 2010, who presented severe urinary incontinence and urethral/bladder neck stricture after prostatic surgery and failure of at least four conservative endoscopic treatments. Six patients underwent a transperineal urethrovesical anastomosis and 11 patients a combined transperineal suprapubical (endoscopic) urethrovesical anastomosis. After six months the patients that presented complete incontinence and no urethral stricture underwent the implantation of an artificial urethral sphincter (AUS). Results. After six months 16 patients were completely incontinent and presented a patent, stable lumen, so that they underwent an AUS implantation. With a mean followup of 50.5 months, 14 patients are perfectly continent with no postvoid residual urine. Conclusions. Two-stage procedures are safe techniques to treat these challenging cases. In our opinion, these cases could be managed with a transperineal approach in patients who present a perfect operative field; on the contrary, in more difficult cases, it would be preferable to use the other technique, with a combined transperineal suprapubical access, to perform a pull-through procedure