Efficacy assessment of different time cycles of nebulized hydrogen peroxide against bacterial and yeast biofilms.

BACKGROUND: The prevention of healthcare-associated infections requires continuous effort. In order to achieve better practical results, the control of environmental microbial biofilms with effective disinfection strategies should be addressed. AIM: To test the efficacy of different time cycles of nebulized hydrogen peroxide (H2O2) against bacterial and yeast dry biofilms. METHODS: The efficacies of a standard cycle (SC) and a fast cycle (FC) of nebulized H2O2 were compared. Microbial biofilms were grown on different material coupons. The metabolic activity of biofilms was determined by XTT assay, and the total biomass of biofilms was determined by crystal violet assay. FINDINGS: Regarding the efficacy of nebulized H2O2 against biofilms, the mean reduction in metabolic activity for the SC was 55.2% [standard deviation (SD) 19.4%], compared with 50.4% (SD 17.7%) for the FC. The mean reduction in total biomass for the SC was 45.5% (SD 22.7%), compared with 46.7% (SD 21.7%) for the FC. No significant differences were found between the tested cycles and materials. CONCLUSION: H2O2 nebulization was found to exhibit good efficacy against healthcare-associated microbial dry biofilms. Moreover, similar efficacies were found between the SC and the FC.

Fast-cycle hydrogen peroxide nebulization against frequent healthcare-associated micro-organisms: efficacy assessment.

BACKGROUND: Hydrogen peroxide (H2O2)-based technology is currently used with the aim of controlling microbial contamination in hospital settings. However, the long cycles required result in prolonged room turnover time, thus precluding a wider implementation of the technology. AIM: To assess the efficacy of a shorter cycle of nebulized H2O2 against healthcare-associated micro-organisms, further comparing among multidrug-resistant and multidrug-susceptible strains. METHODS: The efficacy of a standard cycle (1 h) and of a faster cycle (15 min) of a 7% H2O2 nebulized solution was compared against bacteria and yeasts. MDR and MDS strains were inoculated on polyvinyl chloride, stainless steel, linoleum, napa leather, and formica coupons, and their growth ability was compared. FINDINGS: Globally, the mean efficacy of the standard cycle ranged between 82.5% (±17.0) and 95.9% (±8.3), while the efficacy of the fast cycle ranged between 84.4% (±17.0) and 95.7% (±10.5). No statistically significant differences were found for the majority of the tested cycles and materials. For all the tested strains, no differences were found regarding the efficacy of cycles. CONCLUSION: The very high disinfection efficacy of the fast cycle was found to be similar to that of the standard cycle. Moreover, a similar efficacy was also demonstrated when comparing between multidrug-resistant and multidrug-susceptible strains. This study supports a wider implementation of the technology, with the expected advantages of reducing room turnover time, costs, and indirect infection transmission. Further assessment of the efficacy of this faster cycle against other emergent microbial global threats would be highly recommended.

Eco-friendly non-biocide-release coatings for marine biofouling prevention.

Environmental concerns have been changing the way of looking for solutions to problems. The hydrosphere, together with its biosphere, has been feeling the impact of many pollutants, used for instance in the marine industry for economic reasons or lack of knowledge of their effects. In particular biocides, applied as coatings in paints, are released into the waters becoming toxic and persistent extending their action to an area far beyond the initial coated surface they should protect. In order to minimize these side effects, two biocides, Irgarol (I) and Econea (E), were covalently attached to polyurethane (PU) and foul-release silicone based (PDMS) marine paints through an isocyanate linker. Their antifouling bioactivity was better in PDMS coatings, both for single (Econea) and combined biocides (E/I ratio = 1.5) with contents lower than 0.6 wt%. The treated samples remained almost clean after more than one year immersion in the Portuguese shore of the Atlantic Ocean, and after about 24 weeks under the tropical conditions of Singapore (Fouling rate < 1%). Complementary biofilm adhesion susceptibility tests against Pseudoalteromonas tunicata D2 showed adhesion reduction higher than 90% for PU formulations containing single biocides and close to 100% for PDMS with combined biocides. The eco-toxicity assessment evidenced a low environmental impact, in accordance with the European standards. In addition, shipping field trial tests showed the best antifouling performance for the Econea-based PDMS formulations (E = 0.6 wt%), which remained clean for about nine months in open seawaters, proving the efficacy of this non-release strategy, when applied under dynamic conditions.

Omega-3 therapeutic supplementation in a patient with metastatic adenocarcinoma of the pancreas with muscle mass depletion.

Pancreatic adenocarcinoma has an extremely poor prognosis. With the best available treatments, the median overall survival duration is still less than 1 year. Most patients develop anorexia and major muscle mass loss that interfere with chemotherapy tolerance and survival. In this paper, we present a case in which these problems were a major concern. A multidisciplinary approach with chemotherapy and close nutritional support permitted better control of the disease and longer survival. We also review the literature on nutritional interventions that show an improvement in quality of life and survival in these patients.

Polyethyleneimine and polyethyleneimine-based nanoparticles: novel bacterial and yeast biofilm inhibitors.

Biofilms are commonly involved in medical device-related infections. The purpose of this study was to determine the antimicrobial and anti-biofilm activity of polyethyleneimine (PEI) and PEI-based nanoparticles (nanoPEI) against Staphylococcus aureus, Staphylococcus epidermidis, Acinetobacter baumannii and Candida albicans (clinical and ATCC strains), and to evaluate their effect upon biofilm formation on polyurethane (PUR)-like catheters. MICs and minimal lethal concentrations of PEI and nanoPEI were determined according to CLSI microdilution reference protocols. For PEI, the MIC value was 195.31 mg l(-1) for all the bacteria and 48.83 mg l(-1) for the yeast strains. For nanoPEI, the MIC value was 1250 mg l(-1) for all the strains except A. baumannii, for which it was 2500 mg l(-1). Biofilm formation was assessed with PUR-like catheter segments and biofilm metabolic activity was quantified by colorimetry with a tetrazolium reduction assay. Plasma membrane integrity and membrane potential were assessed by flow cytometry after staining microbial cells with a membrane-impermeable dye, propidium iodide, and a membrane-potential marker, DiBAC4(3). PEI inhibited growth of all microbial species; higher concentrations of nanoPEI were needed to inhibit growth of all species. Biofilm formation in the presence of anti-bacterial PEI activity was dose-dependent (except for S. epidermidis) and species-related. NanoPEI at 0.5×MIC and MIC significantly reduced the metabolic activity of biofilms of S. aureus, S. epidermidis and A. baumannii, whereas 2×MIC was required in order to inhibit biofilm metabolic activity.

Characteristics of cardiac epithelium at the esophagogastric junction of a pediatric population with gastroesophageal reflux.

Cardiac mucosa (CM) of the adult, regardless its location, shares phenotypic characteristics with Barrett's epithelium, namely villin expression and a Barrett's pattern of cytokeratins 7 and 20 expression. As far as we know, the phenotypic profile of CM in children has not been studied. The objective was to evaluate the phenotypic profile of cardiac mucosa from the esophagogastric junction of children with reflux symptoms. Biopsies routinely performed at the esophagogastric junction of children submitted to upper-gastrointestinal endoscopy for complaints suggestive of reflux were retrieved from the archive and used for the purposes of this study. Biopsies were assessed for the presence of squamous epithelium, cardiac and oxyntic mucosa and intestinal metaplasia. Samples displaying both squamous and columnar epithelia were immunohistochemically evaluated for the presence of villin and sucrase-isomaltae and for the expression of cytokeratins 7 and 20. From the 42 biopsies samples retrieved, 30 had simultaneously squamous and columnar epithelia. Cardiac mucosa was present in 86.7% of the cases, and intestinal metaplasia was observed only in one (3.3%). Villin expression in cardiac mucosa was observed in 96% of the cases and a cytokeratins 7 and 20 Barrett's pattern in 73%. Sucrase-isomaltase and MUC2 were only expressed in the case with intestinal metaplasia. Cardiac mucosa was high prevalent in biopsies from the esophagogastric junction of children with reflux. As in adults, cardiac mucosa in children has an immunoprofile similar to Barrett's esophagus. For the first time, it was shown that pediatric cardiac mucosa frequently expresses villin.

Helicobacter pylori virulence genotypes in Portuguese children and adults with gastroduodenal pathology.

The aim of this study was to evaluate the prevalence of virulence genotypes, namely cagA, vacA and babA2, of Helicobacter pylori strains isolated from Portuguese adults and children presenting gastroduodenal pathology. One hundred thirty-six strains were studied, 82 isolated from adult patients (50 with nonulcerative gastritis and 32 with active peptic ulcer) and 58 isolated from children (54 with nonulcerative gastritis and 4 with duodenal ulcer). Genotyping of cagA, vacA and babA2 was assessed by polymerase chain reaction. Overall, Helicobacter pylori strains carrying more virulent genotypes were much more prevalent in adults than in children, particularly the type I ( vacAs1- and cagA-positive) and the triple-positive ( vacAs1-, cagA- and babA2-positive) strains ( P<0.001). A subpopulation of adults and children with nonulcerative gastritis was also studied, and differences in the prevalence of virulent genotypes were observed, either for individual genotypes ( P=0.017 for cagA, P=0.010 for vacAs1) or in combinations, i.e. the type I genotype ( P=0.005) and the triple-positive strains ( P=0.031). There was no difference between the two populations in the distribution of babA2 and m1/m2 genotypes. Considering the cohort effect in the epidemiology of Helicobacter pylori infection, these results suggest that different strains might circulate during different periods of time, or that, after infection in childhood, individual strains will undergo changes during the course of infection.

Features and trends in Helicobacter pylori antibiotic resistance in Lisbon area, Portugal (1990-1999).

The features of Helicobacter pylori antibiotic resistance in Lisbon from 1990 to 1999 were studied. Overall resistance rates to amoxycillin, tetracycline, metronidazole, clarithromycin and ciprofloxacin were 0, 0, 30.6, 19.0 and 9.6%, respectively. The incidence of resistance to clarithromycin was much higher in isolates from children (44.8%) than adults (14.6%). For metronidazole, the contrary was observed (children: 19.0%, adults: 32.3%). Ciprofloxacin-resistant isolates were all from adult patients. Concerning the adult population, the resistance rate to metronidazole showed a slight increase during the decade, while for clarithromycin and ciprofloxacin a significant increase was observed (4.6 to 22.0% and 0 to 20.9%, respectively).

Identification of some volatile descriptors of the rock-rose-like aroma of fortified red wines from douro demarcated region.

2,2,6-Trimethylcyclohexanone and ethyl dihydrocinnamate, known sensorial descriptors of rock-rose (Cistus ladaniferus) aroma, are reported as minor constituents of young fortified wines from the Douro Demarcated region (vintage 1997) for the first time. These compounds were respectively identified by GC-MS in two different fractions (A and B) obtained by preparative GC. Quantitative analysis by GC-MS was performed on 14 fortified wines originating from the different Douro subregions. Ethyl dihydrocinnamate was found to be present at concentrations between 2.3 and 6.7 microgram/L, while 2,2,6-trimethylcyclohexanone ranged from 0.05 to 0.4 microgram/L.

Sodium fluorescein influence on the hemorheological profile of non-insulin dependent diabetes mellitus patients.

Sodium fluorescein angiography is a widely used technology in ophthalmology, which allows us to visualise the chorioretinal microcirculation. Previous reports showed a prolongation of the retinal circulation time along with erythrocyte hyperaggregation and a decrease of erythrocyte acetylcholinesterase activity and a possible interference with the erythrocyte's membrane fluidity. The aim of the present work is to investigate the influence of sodium fluorescein on the hemorheological profile of a group of 23 non-insulin dependent diabetes mellitus (NIDDM) patients undergoing routine retinal angiography. Thirty minutes after the endovenous administration of the fluorescein there was: (I) an increase of whole blood viscosity (p = 0.015), erythrocyte elongation index (EEI, p < 0.05), whole blood pH (p < 0.001), methemoglobin (p < 0.001) and carboxyhemoglobin (p < 0.001) concentrations; (II) no variation of plasma osmolality and erythrocyte aggregation index (EAI); (III) a decrease of the erythrocyte acetylcholinesterase activity, p < 001; (IV) no variation in membrane lipid fluidity, although 1,6-diphenyl-1,3,5-hexatriene (DPH) correlated directly with the EEI, while 1,4-trimethyl-phenyl-1,3,5-hexatriene (TMA-DPH) and EAI correlated inversely, suggesting that the decreasing EEI (lower erythrocyte deformablity) might be associated with an increased rigidity of the external polar region and fluidification of the hydrophobic region of the erythrocyte membrane, with an increasing EAI. In conclusion, the endovenous administration of sodium fluorescein in NIDDM patients during the retinal angiography procedure interferes with the erythrocyte membrane and possibly with the microcirculatory blood flow.

Red blood cell membrane integrity in primary open angle glaucoma: ex vivo and in vitro studies.

PURPOSE: There is increasing evidence that abnormal perfusion of the optic nerve head is an important factor involved in the pathophysiology of glaucoma. Transport and distribution of oxygen to the tissues takes place through the erythrocyte membrane. Red blood cell (RBC) acetylcholinesterase (AChE) is a marker of RBC membrane integrity. The aim of this study was to find out whether RBC membrane integrity is preserved in primary open angle glaucoma (POAG), and whether it is modified by the use of topical timolol maleate and pilocarpine. METHOD: RBC AChE activity was determined ex vivo by Kaplan's spectrophotometric method in 19 POAG patients undergoing topical treatment for glaucoma with timolol, pilocarpine or a combination of the two drugs, and compared with that in 20 controls. To assess the effect of antiglaucomatous therapy in our findings, we carried out an in vitro study in 26 non-glaucomatous patients in which we measured RBC AChE activity after incubation of blood with either timolol maleate, pilocarpine or a combination of the two drugs, using the same spectrophotometric method. RESULTS: There was a significant increase in RBC AChE enzyme activity in POAG patients compared with the control group (p < 0.002). However, timolol and pilocarpine, individually or in combination, have the opposite effect, significantly decreasing RBC AChE activity (p < 0.05). CONCLUSION: This change in RBC AChE enzyme activity could suggest alterations in RBC membrane integrity in primary open angle glaucoma. Whether or not this finding has implications regarding the microcirculation at the optic nerve head needs to be investigated further.

The effect of sodium fluorescein angiography on erythrocyte properties.

Sodium fluorescein angiography is a widely used routine ophthalmological diagnostic procedure which enables the study of chorioretinal microcirculation and consists of the injection of sodium fluorescein into the systemic bloodstream. The aim of the present study was to evaluate whether or not fluorescein interferes with erythrocyte properties during the angiographic procedure. In a group of 37 patients, 26 with non-insulin-dependent diabetes mellitus (DM) with and without retinopathy, and 11 without diabetes mellitus (non-DM) although affected by other ophthalmological diseases, all undergoing routine angiography, blood samples were drawn before (T0) and 30 min (T30) after fluorescein injection. The erythrocyte aggregation index (EAI), membrane lipid fluidity and erythrocyte acetylcholinesterase activity were determined in both groups. After fluorescein injection there was no statistical change in EAI and erythrocyte membrane fluidity in either group. Erythrocyte acetylcholinesterase activity, a marker of membrane protein integrity, decreased significantly (p < 0.01) in the DM group. Membrane lipid fluidity did not change with fluorescein injection, however (i) in the DM group erythrocyte membranes became more rigid than in the non-DM (DPH: p < 0.01); (ii) EAI and membrane lipid fluidity became significantly correlated (r = 0.6263, p < 0.05) in non-DM patients at T30. In conclusion, fluorescein administration for angiographic procedures seems to interact with erythrocyte membrane, namely, in diabetic patients, which may interfere with the blood flow in the microcirculation.

[The effect of angiography with disodium fluorescein on the hemorheologic parameters of diabetics]. / Estudo da influência da angiografia com fluoresceína bissódica nos parâmetros hemorreológicos de doentes com diabetes mellitus.

Chorioretinal angiography with fluorescein is an auxiliary method widely used in Ophthalmology which enables us to study the blood/humour and blood/retina/choroid interface. The presence of a strange compound in circulation may interfere with blood flow homeostasis. With the aim of studying fluorescein influence in blood rheology, blood samples were drawn before, immediately after and 30 minutes after fluorescein bolus injection; from 10 patients with diabetes mellitus, with or without retinopathy, blood fluidity (hematocrit, erythrocyte aggregation and whole blood viscosity) and erythrocyte membrane functional properties were determined. After fluorescein injection there was erythrocyte hyperaggregation (37.7%, p = 0.004), was detected, hemoglobin concentration (p = 0.039) and hematocrit (p = 0.013) decrease, and a double time increase of methemoglobin concentration (p < 0.001) the erythrocyte membrane hydrophobic region became more rigid (p = 0.016). All these abnormalities normalized after 30 minutes. In conclusion, fluorescein angiography interferes acutely with the hemorheological parameters of patients with diabetes mellitus, with erythrocyte hyperaggregation which could interfere with the microcirculation of these patients.

[Value of congenital hypertrophy of the retinal pigment epithelium as diagnostic marker in familial adenomatous polyposis]. / Valor da hipertrofia congénita do epitélio pigmentar da retina como marcador diagnóstico na polipose adenomatosa familiar.

Familial Adenomatous Polyposis (FAP) is a dominant autosomic disease characterized by the development of hundreds to thousands of colonic adenomatous polyps. Affected patients have a 100% risk of colon cancer development if they are not submitted to a prophylactic colectomy. Identification of carriers depends on the detection of colonic polyps, and endoscopic surveillance must be offered to all descendents, including healthy individuals. Congenital hypertrophy of the retinal pigment epithelium (CHRPE) has been suggested to have a correlation with FAP trait, even before colonic polyp development. The objective of this study is to evaluate CHRPE as a diagnostic marker in FAP patients and descendents. CHRPE was studied in 26 members of 7 FAP families, using direct and indirect ophthalmoscopy, biomicroscopy and retinography. It was found in 62.5% of patients and in 10% of the descendents at risk. Two families did not show signs of CHRPE. Affected members in the remaining families, had positive examinations in 83.3% (two affected members were negative). These results suggest that CHRPE is an important diagnostic tool to identify FAP patients in those families which express the marker. To those descendents who have negative examinations, whether they belong to positive or negative CHRPE families, identification of FAP trait depends on endoscopic surveillance in order to detect colonic polyps.

Behçet's disease in Portugal.

During the last decades 156 patients with Behçet's disease were diagnosed and studied in Portugal. Until 1966 only two cases were referred in Portugal. In 1978 we reported 23 cases (18 males and 5 females) with multisystemic involvement, not different from that reported in other publications elsewhere. In 1987, 17 cases were reported from the Oporto region, diagnosed during a period of 5 years. In the region of Coimbra 15 cases were diagnosed (6 males and 9 females). In the Lisbon area from 1969 until 1990, 123 cases were studied altogether, 76 during the last decade (35 males and 41 females). All the cases studied had oral ulcerations and a high incidence of ocular involvement (clinical and sub-clinical, 87%). HLA-B5 was detected in 50% of the patients. At least four cases were fatal, three died with neuro-Behçet. Steroids, colchicine and thalidomide were the main form of treatment, for the patients in the Lisbon area, but in some cases chlorambucil, cyclophosphamide, azathioprine, cyclosporine-A, fibrinolytic therapy, plasmapheresis and isovolemic hemodilution were also used.

Reaction of rectal mucosa of celiac patients to direct contact with gluten.

It is assumed that the colonic mucosa of celiac patients is not sensitive to gluten. This assumption has been supported by the absence of clinical manifestations of colonic involvement in patients with active celiac disease which, by itself, does not confirm insensitivity to gluten. Eight children, aged from 11 to 25 months, with an initial diagnosis of celiac disease were studied: in five children a definite diagnosis has already been confirmed. Rectal gluten challenge was done by means of retention enemas. A volume of 100 ml of a 10% gluten suspension in water was introduced into the rectum three times per day for 8 days; each enema was retained at least 1 h. Rectosigmoidoscopies and rectal biopsies were done before and at the end of the challenge period. The endoscopic appearance of rectal mucosa was normal in all the children either before or after gluten challenge. The means of total mucosal thickness, intraepithelial lymphocyte counts, mitotic crypt cell activity, and cellular infiltration of lamina propria increased after challenge; the mean of goblet cell/epithelial cell ratio in the surface epithelium decreased. The differences between each pair of means (before and after challenge), however, were not significant except for total mucosal thickness (p less than 0.05), the meaning of which is unclear. This study did not definitely demonstrate that the rectal mucosa of celiacs is insensitive to gluten. For practical clinical purposes, however, it behaves as such. This makes the rectal mucosa a useless tool for the final diagnosis of celiac disease.