Rhamnolipid 89 Biosurfactant Is Effective against Streptococcus oralis Biofilm and Preserves Osteoblast Behavior: Perspectives in Dental Implantology.

Biofilm-related peri-implant diseases represent the major complication for osteointegrated dental implants, requiring complex treatments or implant removal. Microbial biosurfactants emerged as new antibiofilm coating agents for implantable devices thanks to their high biocompatibility. This study aimed to assess the efficacy of the rhamnolipid 89 biosurfactant (R89BS) in limiting Streptococcus oralis biofilm formation and dislodging sessile cells from medical grade titanium, but preserving adhesion and proliferation of human osteoblasts. The inhibitory activity of a R89BS coating on S. oralis biofilm formation was assayed by quantifying biofilm biomass and microbial cells on titanium discs incubated up to 72 h. R89BS dispersal activity was addressed by measuring residual biomass of pre-formed biofilms after rhamnolipid treatment up to 24 h. Adhesion and proliferation of human primary osteoblasts on R89BS-coated titanium were evaluated by cell count and adenosine-triphosphate quantification, while cell differentiation was studied by measuring alkaline phosphatase activity and observing mineral deposition. Results showed that R89BS coating inhibited S. oralis biofilm formation by 80% at 72 h and dislodged 63-86% of pre-formed biofilms in 24 h according to concentration. No change in the adhesion of human osteoblasts was observed, whereas proliferation was reduced accompanied by an increase in cell differentiation. R89BS effectively counteracts S. oralis biofilm formation on titanium and preserves overall osteoblasts behavior representing a promising preventive strategy against biofilm-related peri-implant diseases.

Rhamnolipid coating reduces microbial biofilm formation on titanium implants: an in vitro study.

BACKGROUND: Peri-implant mucositis and peri-implantitis are biofilm-related diseases causing major concern in oral implantology, requiring complex anti-infective procedures or implant removal. Microbial biosurfactants emerged as new anti-biofilm agents for coating implantable devices preserving biocompatibility. This study aimed to assess the efficacy of rhamnolipid biosurfactant R89 (R89BS) to reduce Staphylococcus aureus and Staphylococcus epidermidis biofilm formation on titanium. METHODS: R89BS was physically adsorbed on titanium discs (TDs). Cytotoxicity of coated TDs was evaluated on normal lung fibroblasts (MRC5) using a lactate dehydrogenase assay. The ability of coated TDs to inhibit biofilm formation was evaluated by quantifying biofilm biomass and cell metabolic activity, at different time-points, with respect to uncoated controls. A qualitative analysis of sessile bacteria was also performed by scanning electron microscopy. RESULTS: R89BS-coated discs showed no cytotoxic effects. TDs coated with 4 mg/mL R89BS inhibited the biofilm biomass of S. aureus by 99%, 47% and 7% and of S. epidermidis by 54%, 29%, and 10% at 24, 48 and 72 h respectively. A significant reduction of the biofilm metabolic activity was also documented. The same coating applied on three commercial implant surfaces resulted in a biomass inhibition higher than 90% for S. aureus, and up to 78% for S. epidermidis at 24 h. CONCLUSIONS: R89BS-coating was effective in reducing Staphylococcus biofilm formation at the titanium implant surface. The anti-biofilm action can be obtained on several different commercially available implant surfaces, independently of their surface morphology.

Medical-Grade Silicone Coated with Rhamnolipid R89 Is Effective against Staphylococcus spp. Biofilms.

Staphylococcus aureus and Staphylococcus epidermidis are considered two of the most important pathogens, and their biofilms frequently cause device-associated infections. Microbial biosurfactants recently emerged as a new generation of anti-adhesive and anti-biofilm agents for coating implantable devices to preserve biocompatibility. In this study, R89 biosurfactant (R89BS) was evaluated as an anti-biofilm coating on medical-grade silicone. R89BS is composed of homologues of the mono- (75%) and di-rhamnolipid (25%) families, as evidenced by mass spectrometry analysis. The antimicrobial activity against Staphylococcus spp. planktonic and sessile cells was evaluated by microdilution and metabolic activity assays. R89BS inhibited S. aureus and S. epidermidis growth with minimal inhibitory concentrations (MIC99) of 0.06 and 0.12 mg/mL, respectively and dispersed their pre-formed biofilms up to 93%. Silicone elastomeric discs (SEDs) coated by R89BS simple adsorption significantly counteracted Staphylococcus spp. biofilm formation, in terms of both built-up biomass (up to 60% inhibition at 72 h) and cell metabolic activity (up to 68% inhibition at 72 h). SEM analysis revealed significant inhibition of the amount of biofilm-covered surface. No cytotoxic effect on eukaryotic cells was detected at concentrations up to 0.2 mg/mL. R89BS-coated SEDs satisfy biocompatibility requirements for leaching products. Results indicate that rhamnolipid coatings are effective anti-biofilm treatments and represent a promising strategy for the prevention of infection associated with implantable devices.

Early healing of peri-implant mucosa in man.

AIM: To analyse (i) cellular and vascular densities in the connective tissue interface portion of the peri-implant mucosa and (ii) tissue interactions with the titanium surface during early stages of healing. MATERIALS AND METHODS: Circumferential biopsies of peri-implant soft tissues were retrieved together with custom-made abutments at 27 implants in 21 patients after 2, 4, 6, 8 and 12 weeks of healing. Following fixation, the peri-implant soft tissue was separated from the abutments, divided into four units and embedded in paraffin. Sections were produced and prepared for immunohistochemical analysis. The abutments were examined by SEM. RESULTS: T and B cells occurred in clusters with a decreasing cell density from 4 to 8 weeks of healing in the connective tissue lateral of the abutment. Macrophages were evenly distributed in the connective tissue along the abutment/tissue interface, while polymorphonuclear (PMN) cells were confined to the tissue portion lateral to the junctional epithelium. Vascular structures showed a decrease in density from 2 to 8 weeks of healing. SEM analyses of the abutments revealed an increased presence of tissue remnants attached to the surface with increasing healing time. A biofilm was consistently observed in a supra-mucosal position, apical of which a "clear zone" occurred that separated the tissue remnants and the biofilm. CONCLUSION: Onset and resolution of inflammation together with increasing tissue attachment to the implant characterize healing of peri-implant mucosa.

Solidago, orthosiphon, birch and cranberry extracts can decrease microbial colonization and biofilm development in indwelling urinary catheter: a microbiologic and ultrastructural pilot study.

PURPOSE: Plants extracts are used in urology to manage urinary tract infections. We aimed to evaluate the efficacy of a preparation with solidago, orthosiphon, birch and cranberry extracts (CISTIMEV PLUS(®)) in reducing microbial colonization and biofilm development in patients with indwelling urinary catheters. METHODS: All consecutive outpatients attending our department between January and June 2010 for the substitution of indwelling catheters were considered for this single-blinded, randomized and controlled pilot study to test superiority of the preventative management (CISTIMEV PLUS(®), 1 tablet daily for 30 days) in respect to no treatment. A sample size of 10-40 participants per group was considered adequate. All patients underwent urine culture the same day of the catheter substitution and were then randomized into test group (n = 48) and control group (n = 35). Ultrastructural analysis was also performed. After 30 days, the catheter was replaced and the analysis repeated. The primary outcome was the rate of positive urinary culture at the end of the entire study period. RESULTS: Ten patients abandoned the study. At 30 days, according to per-protocol analysis, the groups statistically differed regarding the rate of positive urine cultures: test group 10/43 and control group 16/30 (p = 0.013) (-30.1 % [95 % CI -51.94 to -8.21]). The most common isolated bacteria were Escherichia coli and Enterococcus faecalis. CONCLUSIONS: The use of solidago, orthosiphon, birch and cranberry extracts resulted in a significant reduction of microbial colonization in patients with indwelling urinary catheters. Larger clinical trials are needed to demonstrate that the effects here reported are sufficient to reduce symptomatic catheter-associated urinary tract infections.

Morphogenesis of peri-implant mucosa revisited: an experimental study in humans.

AIM: To apply a novel human model to evaluate the morphogenesis of the mucosal attachment to implants. MATERIAL AND METHODS: Twenty one patients receiving implant-supported single-tooth replacement were enrolled in this study. After implant installation, a custom-designed experimental abutment was connected to the implant. Soft tissue biopsies representing 2, 4, 8 or 12 weeks of healing were collected by the use of a circular cutting device and prepared for histological analysis. RESULTS: The soft tissue biopsies were retrieved, preserved and processed with a technique that was safe and reproducible. The results from the histological analysis in regards to dimensional and qualitative changes in the mucosa over time were consistent with those reported from animal experiments. At 8 weeks, the soft tissue dimension was about 3.6 mm and included a barrier epithelium of 1.9 mm and a connective tissue portion of 1.7 mm. Similar dimensions were found at 12 weeks. CONCLUSION: It is suggested that the new human model provides advantages in terms of cost-effectiveness in research as well as from ethical aspects and should be considered as an alternative to pre-clinical in vivo studies in animals.

Blood flow in hemodialysis catheters: a numerical simulation and microscopic analysis of in vivo-formed fibrin.

Although catheters with side holes allow high flow rate during hemodialysis, they also induce flow disturbances and create a critical hemodynamic environment that can favor fibrin deposition and thrombus formation. This study compared the blood flow and analyzed the influence of shear stress and shear rate in fibrin deposition and thrombus formation in nontunneled hemodialysis catheters with unobstructed side holes (unobstructed device) or with some side holes obstructed by blood thrombi (obstructed device). Computational fluid dynamics (CFD) was performed to simulate realistic blood flow under laminar and turbulent conditions. The results from the numerical simulations were compared with the fibrin distribution and thrombus architecture data obtained from scanning electron microscopy (SEM) and two photons laser scanning microscopy (TPLSM) on human thrombus formed in catheters removed from patients. CFD showed that regions of flow eddies and separation were mainly found in the venous holes region. TPLSM characterization of thrombi and fibrin structure in patient samples showed fibrin formations in accordance with simulated flux dynamics. Under laminar flow conditions, the wall shear stress close to border holes increased from 87.3±0.2 Pa in the unobstructed device to 176.2±0.5 Pa in the obstructed one. Under turbulent flow conditions, the shear stress increased by 47% when comparing the obstructed to the unobstructed catheter. The shear rates were generally higher than 5000/s and therefore sufficient to induce fibrin deposition. This findings were supported by SEM data documenting a preferential fibrin arrangement on side hole walls.

Effect of chlorhexidine in preventing plaque biofilm on healing abutment: a crossover controlled study.

INTRODUCTION: The study aimed at evaluating the effect of chlorhexidine (CHX) in preventing plaque biofilm (PB) formation on healing abutments (HAs) in patients rehabilitated with osseointegrated implants. MATERIALS AND METHODS: Fifty HAs were placed in 34 voluntary patients 1 week after implant surgery (test group). After 7 days, a new set of 50 HAs was placed in the same implant sites and removed 1 week after (control group). During the 2 testing periods, patients were instructed to apply: CHX mouth rinsing twice daily and no brushing (test); no CHX mouth rinsing and no brushing (control). Scanning electron microscopy and image analysis were blindly used to objectively quantify PB amount on removed HAs. RESULTS: Median values and interquartile ranges of the percent ratio of titanium surface covered from PB were 0.9 (0.1-4.1) and 1.2 (0.1-11.6) for test and control groups, respectively (P = 0.0275). CONCLUSIONS: CHX mouth rinsing significantly limited plaque formation on HAs, being a valid contribution to mechanical brushing in early phases of plaque control on dental implants.

Effects of mushroom and chicory extracts on the shape, physiology and proteome of the cariogenic bacterium Streptococcus mutans.

BACKGROUND: Dental caries is an infectious disease which results from the acidic demineralisation of the tooth enamel and dentine as a consequence of the dental plaque (a microbial biofilm) accumulation. Research showed that several foods contain some components with antibacterial and antiplaque activity. Previous studies indicated antimicrobial and antiplaque activities in a low-molecular-mass (LMM) fraction of extracts from either an edible mushroom (Lentinus edodes) or from Italian red chicory (Cichorium intybus). METHODS: We have evaluated the antimicrobial mode of action of these fractions on Streptococcus mutans, the etiological agent of human dental caries. The effects on shape, macromolecular syntheses and cell proteome were analysed. RESULTS: The best antimicrobial activity has been displayed by the LMM mushroom extract with a bacteriostatic effect. At the MIC of both extracts DNA synthesis was the main macromolecular synthesis inhibited, RNA synthesis was less inhibited than that of DNA and protein synthesis was inhibited only by roughly 50%. The partial inhibition of protein synthesis is compatible with the observed significant increase in cell mass. The increase in these parameters is linked to the morphological alteration with transition from cocci of the untreated control to elongated cells. Interestingly, these modifications were also observed at sub-MIC concentrations. Finally, membrane and cytosol proteome analysis was conducted under LMM mushroom extract treatment in comparison with untreated S. mutans cells. Significant changes were observed for 31 membrane proteins and 20 of the cytosol fractions. The possible role of the changed proteins is discussed. CONCLUSIONS: This report has shown an antibiotic-like mode of action of mushroom and chicory extracts as demonstrated by induced morphogenetic effects and inhibition of specific macromolecular synthesis. This feature as well as the safe use of this extract as result of its natural origin render the LMM both mushroom and chicory extracts suitable for the formulation into products for daily oral hygiene such as mouthwashes or toothpastes.

Staphylococcus epidermidis is a safer surrogate of Staphylococcus aureus in testing bacterial filtration efficiency of face masks.

Face masks play a role in reducing the spread of airborne pathogens, providing that they have a good filtration performance, are correctly fitted and maintained. Bacterial Filtration Efficiency (BFE) is a key indicator for evaluating filtration performance according to both European and US standards, requiring the use of Staphylococcus aureus loaded aerosol. However, the generation and handling of a Biohazard group 2 bacterium aerosol require a careful management of the biological risk and pose limitations to the accessibility to this method. To mitigate these drawbacks, we investigated the use of S. epidermidis ATCC 12228, a Biohazard group 1 bacterium, as surrogate in BFE test. To this end, tests with the surrogate strain were performed to tune the method. Then, three face mask models, representative for both surgical and community masks, were tested according to the standard method and then using an aerosolized suspension of S. epidermidis. BFE% values were calculated for each mask model and tested microorganisms. Results showed that BFE test can be performed using the S. epidermidis instead of S. aureus, preserving results validity and turnaround time, but reducing residual risk for laboratory operators.

Unusual case of necrotizing fasciitis caused by Vibrio cholerae O137.

We report a case of necrotizing fasciitis caused by Vibrio cholerae O137 in an immunocompromised 49-year-old man. The infection was acquired following a minor traumatic injury and exposure to seawater during the summer of 2009 in Italy. Although highly immunocompromised, the patient survived. The strain was cytotoxic, invasive, and adhesive and contained a fragment of the El Tor-like hemolysin (El Tor hlyA) gene.

Effects of mushroom and chicory extracts on the physiology and shape of Prevotella intermedia, a periodontopathogenic bacterium.

Contrary to the common assumption that food has a negative impact on oral health, research has shown that several foods contain a number of components with antibacterial and antiplaque activity. These natural compounds may be useful for improving daily oral hygiene. In this study we evaluate the mode of antimicrobial action of fractions of mushroom and red chicory extracts on Prevotella intermedia, a periodontopathogenic bacterium. The minimal inhibitory concentration corresponded to 0.5x compared to the natural food concentration for both extracts. This concentration resulted in a bacteriostatic effect in mushroom extract and in a slightly bactericidal effect in chicory extract. Cell mass continued to increase even after division stopped. As regards macromolecular synthesis, DNA was almost totally inhibited upon addition of either mushroom or chicory extract, and RNA to a lesser extent, while protein synthesis continued. Cell elongation occurred after septum inhibition as documented by scanning electron microscopy and cell measurement. The morphogenetic effects are reminiscent of the mode of action of antibiotics such as quinolones or ß-lactams. The discovery of an antibiotic-like mode of action suggests that these extracts can be advantageously employed for daily oral hygiene in formulations of cosmetic products such as mouthwashes and toothpastes.

Counter-Acting Candida albicans-Staphylococcus aureus Mixed Biofilm on Titanium Implants Using Microbial Biosurfactants.

This study aimed to grow a fungal-bacterial mixed biofilm on medical-grade titanium and assess the ability of the biosurfactant R89 (R89BS) coating to inhibit biofilm formation. Coated titanium discs (TDs) were obtained by physical absorption of R89BS. Candida albicans-Staphylococcus aureus biofilm on TDs was grown in Yeast Nitrogen Base, supplemented with dextrose and fetal bovine serum, renewing growth medium every 24 h and incubating at 37 °C under agitation. The anti-biofilm activity was evaluated by quantifying total biomass, microbial metabolic activity and microbial viability at 24, 48, and 72 h on coated and uncoated TDs. Scanning electron microscopy was used to evaluate biofilm architecture. R89BS cytotoxicity on human primary osteoblasts was assayed on solutions at concentrations from 0 to 200 µg/mL and using eluates from coated TDs. Mixed biofilm was significantly inhibited by R89BS coating, with similar effects on biofilm biomass, cell metabolic activity and cell viability. A biofilm inhibition >90% was observed at 24 h. A lower but significant inhibition was still present at 48 h of incubation. Viability tests on primary osteoblasts showed no cytotoxicity of coated TDs. R89BS coating was effective in reducing C. albicans-S. aureus mixed biofilm on titanium surfaces and is a promising strategy to prevent dental implants microbial colonization.

Prostate calcifications: A case series supporting the microbial biofilm theory.

Purpose: Prostate calcifications are a common finding during transrectal prostate ultrasound in both healthy subjects and patients, but their etiopathogenesis and clinical significance are not fully understood. We aimed to establish a new methodology for evaluating the role of microbial biofilms in the genesis of prostate calcifications. Materials and Methods: Ten consecutive patients who had undergone radical prostatectomy were enrolled in this study. All of the patients presented with prostate calcifications during transrectal ultrasound evaluation before surgery and underwent Meares-Stamey tests and clinical evaluation with the National Institutes of Health Chronic Prostatitis Symptom Index and the International Prostate Symptom Score. At the time of radical prostatectomy, the prostate specimen, after removal, was analyzed with ultrasonography under sterile conditions in the operating room. Core biopsy specimens were taken from the site of prostate calcification and subjected to ultrastructural and microbiological analysis. Results: The results of the Meares-Stamey test showed only 1 of 10 patients (10%) with positive cultures for Escherichia coli. Two of five patients (40%) had positive cultures from prostate biopsy specimens. Enterococcus faecalis, Enterococcus raffinosus, and Citrobacter freundii were isolated. Ultrastructural analysis of the prostate biopsy specimens showed prostate calcifications in 6 of 10 patients (60%), and a structured microbial biofilm in 1 patient who had positive cultures for E. faecalis and E. raffinosus. Conclusions: Although the findings are supported by a low number of patients, this study highlights the validity of the proposed methodology for investigating the role of bacterial biofilms in the genesis of prostate calcification.

Sterility and microbiological assessment of reused single-use cardiac electrophysiology catheters.

OBJECTIVE: To assess the performance and limitations of a reprocessing protocol for nonlumen electrophysiology catheters by testing the sterility of reprocessed devices and defining the maximum number of reprocessing cycles sustainable by the device in hygienically safe conditions. DESIGN: Simulated use, reprocessing, and testing of the catheters. SETTING: Microbiology and virology department of a public health diagnostic laboratory. INTERVENTIONS: Seventy-three catheters were collected after clinical use on patients. The first group of devices was tested for sterility after 1 cycle of reprocessing. By the repetition of simulated use (blood inoculated with bacteria) and reprocessing (decontamination, cleaning, and hydrogen peroxide gas plasma sterilization), we obtained 39 sample devices reprocessed 2 times, 26 reprocessed 3 times, 28 reprocessed 4 times, 36 reprocessed 5 times, and 22 reprocessed 6 times. Devices were cultured for 28 days in trypticase soy broth. RESULTS: We tested 208 catheters with 6 cycles of reprocessing and 4 inoculated bacteria species. No devices tested positive for the inoculated strains until the fourth cycle of reprocessing. One of 35 catheters showed the growth of the inoculated strain Bacillus subtilis after 5 cycles of reprocessing, and 1 of 22 catheters showed growth of this organism 6 cycles. After the second reprocessing, 7 of 36 devices showed growth of gram-negative bacteria other than the strain inoculated. CONCLUSIONS: Reprocessing according to the reprocessing protocol was insufficient to guarantee device sterility after 5 reuses. Cleaning with enzymatic solution revealed good cleaning properties with efficient bioburden reduction. Storage intervals of longer than 24 hours during reprocessing should be avoided to limit contamination or bacterial overgrowth. Technical considerations suggest the introduction of reprocessing procedures only in hospitals with considerable workloads.

Efficiency in endotoxin removal by a reprocessing protocol for electrophysiology catheters based on hydrogen peroxide plasma sterilization.

Electrophysiology and ablation cardiac catheters, which come in contact with blood during clinical use, are required to be non-pyrogenic (<20 endotoxin units (EU)/device). This study aimed to quantify the residual endotoxin load in reprocessed devices as a mandatory step to guarantee safe reuse. We monitored the pyrogenic status of the device (n=61) in three fundamental steps of the reprocessing protocol: after clinical use, after decontamination-cleaning treatments and after complete reprocessing, including sterilization by hydrogen peroxide gas plasma. Finally, a depyrogenation test was produced for evaluating the depyrogenation efficiency of the sole hydrogen peroxide sterilization treatment. Results showed that standard clinical use did not represent a source for endotoxin contamination, while the use of tap water and manual cleaning processing could increase the pyrogenic load in a significant way. The introduction of the sterilization by hydrogen peroxide gas plasma resulted in effective reduction of the endotoxin contamination and in safe reprocessing of 15 of 15 clinically used catheters. In addition, tests conducted on in vitro spiked catheters showed that initial pyrogenic loads of 40, 80, 200EU/device were reduced to less than 11EU/device. Depyrogenation testing demonstrated efficiency in endotoxin reduction of more than 62 times (1.8log). These results show the determining role of hydrogen peroxide gas-plasma sterilization in the reduction of pyrogenic load on medical devices. Considering actual hygienic requirements at single-use device reprocessing, hydrogen peroxide gas-plasma sterilization can be considered as an efficient treatment at non-lumen cardiac electrophysiology catheter reprocessing.

A Multi-techniques Approach to assess reprocessing of Single-use electrosurgical pencils.

Reprocessing and reuse of single-use electrosurgical pencils (EPs) is a diffused practice in countries with limited access to healthcare subvention system and in developing countries. However, safety and efficacy issues are associated to this practice, requiring specific methods for checking the reprocessed device before clinical re-use. This study aimed at defining a set of testing methods for assessing thermal and surface characteristics of reprocessed single-use EPs and evaluating the suitability of these techniques for revealing modifications between brand new and reprocessed single-use EPs. We reported a multi-technique approach based on optical and electron microscopy, X-rays spectroscopy and thermal analysis. The assessment of a total of 30 variables of interest on both brand new and reprocessed devices, allowed to identify the most informative ones. Seven of the evaluated variables were found to differentiate the reprocessed device from the new ones in a significant way. The presented methods deserve potential for tracking modifications during the device lifecycle.

A spectral approach for the quantitative description of cardiac collagen network from nonlinear optical imaging.

The assessment of collagen structure in cardiac pathology, such as atrial fibrillation (AF), is essential for a complete understanding of the disease. This paper introduces a novel methodology for the quantitative description of collagen network properties, based on the combination of nonlinear optical microscopy with a spectral approach of image processing and analysis. Second-harmonic generation (SHG) microscopy was applied to atrial tissue samples from cardiac surgery patients, providing label-free, selective visualization of the collagen structure. The spectral analysis framework, based on 2D-FFT, was applied to the SHG images, yielding a multiparametric description of collagen fiber orientation (angle and anisotropy indexes) and texture scale (dominant wavelength and peak dispersion indexes). The proof-of-concept application of the methodology showed the capability of our approach to detect and quantify differences in the structural properties of the collagen network in AF versus sinus rhythm patients. These results suggest the potential of our approach in the assessment of collagen properties in cardiac pathologies related to a fibrotic structural component.

Quantification of fibrin in blood thrombi formed in hemodialysis central venous catheters: a pilot study on 43 CVCs.

PURPOSE: Fibrin deposition and thrombotic occlusion represent a serious cause of access dysfunction in hemodialysis central venous catheters (CVCs). The aim of this work was to define and apply a method for imaging and quantifying fibrin in thrombi formed into the side holes of CVCs. METHODS: Forty-three CVCs removed from a cohort of dialyzed patients were analyzed in this pilot study. Hematoxylin and eosin and a modified Carstair's staining were applied on permanent thrombus sections. Fluorescence microscopy and image analysis were performed to quantify the fibrin amount. RESULTS: Highly fluorescent areas were invariably associated with fibrin by Carstair's method. The deposition of concentric layers of fibrin and erythrocytes was easily identified by fluorescence microscopy, showing growth features of the thrombus. Fibrin amount in diabetic patients was significantly higher than that in nondiabetic patients with median (interquartile range) values of 51% (47-68%) and 44% (30-54%), respectively (p=0.032). No significant difference in fibrin content was found by grouping data according to catheter type, permanence time, insertion site and dialysis vintage. Higher variability in fibrin values was found in thrombi from CVCs removed after 1-15 days compared with 16-60 days. A trend of an increase in fibrin amount in thrombi was noted according to blood platelet count at CVC insertion. CONCLUSIONS: The analytical method presented here proved to be a rapid and effective way for quantifying fibrin content in thrombi formed on CVCs with potential application in future clinical studies.

Effects of pollen extract in association with vitamins (DEPROX 500®) for pain relief in patients affected by chronic prostatitis/chronic pelvic pain syndrome: results from a pilot study.

BACKGROUND: The therapeutic efficacy of CP/CPPS is not very satisfactory and the impact on young male's quality of life is considerable. The aim of the present study is to evaluate the efficacy of pollen extract associated with vitamins (DEPROX 500®) in order to improve the quality of life of young patients affected by chronic prostatitis type IIIb (CP/CPPS) by pain relieving. METHODS: All patients with clinical and instrumental diagnosis of CP/CPPS (class b) underwent DEPROX 500® 2 tablets in a single dose daily for 30 days. Clinical and microbiological analyses were carried out at the enrolment and after 1 month. NIH-CPSI and IPSS questionnaires have been used. The main outcome measure was the improvement of quality of life at the end of the whole study period, evaluated by questionnaires results. RESULTS: 20 men (mean age 32.8 ± 6.78) were enrolled in this pilot study. The baseline questionnaire mean scores were 25.90 ± 2.1 and 8.01 ± 3.64 for NIH-CPSI and IPSS, respectively. At the follow-up examination (1 month after treatment), 18 out of 20 patients (90.0%) reported an improvement of quality of life, in terms of pain reduction. The questionnaire results after 1 month from treatment were as follows: NIH-CPSI 12.8 ± 2.20, IPSS 7.6 ± 1.58. Statistically significant differences were then reported between the two visits, in terms of NIH-CPSI scores (p<0.001). No statistically significant differences have been reported in terms of IPSS between the two groups. All patients were negative at the Meares-Stamey test evaluation. The compliance to the study protocol was 100%. CONCLUSIONS: The pollen extract associated with vitamins (DEPROX 500®) significantly improved total symptoms, pain, and QoL in patients with non-inflammatory CP/CPPS without severe side effects.