Light-induced antibacterial activity of electrospun chitosan-based material containing photosensitizer.

Increasing antimicrobial resistance requires the development of novel materials and approaches for treatment of various infections. Utilization of photodynamic therapy represents an advanced alternative to antibiotics and metal-based agents. Here, we report the fabrication of electrospun material that possesses benefits of both topical antimicrobial and photodynamic therapies. This material combines chitosan, as a biocompatible polymer, and a second generation photosensitizer. The incorporation of photosensitizer doesn't affect the material morphology and its nearly uniform distribution in fibers structure was observed by confocal Raman microscopy. Owing to photosensitizer the prepared material exhibits the light-induced and spatially limited antimicrobial activity that was demonstrated against Staphylococcus aureus, an important etiological infectious agent. Such material can be potentially used in antibacterial therapy of chronic wounds, infections of diabetic ulcers, and burns, as well as rapidly spreading and intractable soft-tissue infections caused by resistant bacteria.

Photosensitizer-loaded electrospun chitosan-based scaffolds for photodynamic therapy and tissue engineering.

Novel chitosan-based nanofibrous composite materials containing different amounts of the photosensitizer Photosens were obtained by electrospinning and were characterized by scanning electron microscopy and by confocal laser scanning microscopy. The release of Photosens from the materials was investigated in water and in phosphate-buffered saline. A noncancerous (MC3T3-E1 murine osteoblasts) and a cancerous [T-47D (mammary gland)] cell line were cultivated on Photosens-containing scaffolds, and cell growth and metabolic activity were examined by confocal laser scanning microscopy and by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphe-nyltetrazolium bromide assay, respectively. The viability of both cell lines on Photosens-containing fibers decreased in a spatial manner upon laser irradiation of an appropriate wavelength and power density. Interestingly, the noncancerous MC3T3-E1 cells grown on Photosens -containing scaffolds were less affected by the irradiation. We conclude that the Photosens-containing electrospun chitosan nanofibers described here are of potential interest for biomedical applications, particularly topical photodynamic therapy and tissue engineering.

Relationship between pathogenic, clinical, and virulence factors of Staphylococcus aureus in infective endocarditis versus uncomplicated bacteremia: a case-control study.

Pathogenic factors of Staphylococcus aureus (SA) in the development of infective endocarditis (IE) have not been sufficiently investigated. The purpose of this study was to analyze the pathogenesis and virulence factors of SA in patients with IE as compared to patients with uncomplicated bacteremia (un-BAC). This is a retrospective case-control study (2002-2014) performed at a tertiary hospital in Spain. Clinical and epidemiological factors were analyzed. We assessed the presence of toxin genes [toxic shock syndrome toxin 1 (tst-1) and enterotoxins A (etA), B (etB), and D (etD)] and the potential relationship between accessory gene regulator (agr) groups and the development of IE confirmed by polymerase chain reaction (PCR). Twenty-nine patients with IE were compared with 58 patients with uncomplicated S. aureus bacteremia (SAB). As many as 75.9 % of patients had community-acquired IE (p < 0.005). Multivariate analysis revealed that there is a significant relationship between community-acquired infection and severe sepsis or septic shock and IE. Also, a minimum inhibitory concentration (MIC) of vancomycin ≥1.5 µg/ml was found to be associated with IE. The agr group I was prevalent (55.2 % vs. 31.0 %; p = 0.030). No association was observed between toxin genes (tst-1, etA, etB, and etD) and IE. The superantigen (SAg) most frequently found in SA isolates was tst-1 (12.6 %). We found no association between toxin genes and IE, probably due to the small sample size. However, a direct relationship was found between agr I and the development of IE, which suggests that agr I strains may have more potential to cause IE.

Left-sided infective endocarditis in patients with liver cirrhosis.

OBJECTIVE: To evaluate the course of left-sided infective endocarditis (LsIE) in patients with liver cirrhosis (LC) analyzing its influence on mortality and the impact of surgery. METHODS: Prospective cohort study, conducted from 1984 to 2013 in 26 Spanish hospitals. RESULTS: A total of 3.136 patients with LsIE were enrolled and 308 had LC: 151 Child-Pugh A, 103 B, 34 C and 20 were excluded because of unknown stage. Mortality was significantly higher in the patients with LsIE and LC (42.5% vs. 28.4%; p < 0.01) and this condition was in general an independent worse factor for outcome (HR 1.51, 95% CI: 1.23-1.85; p < 0.001). However, patients in stage A had similar mortality to patients without cirrhosis (31.8% vs. 28.4% p = NS) and in this stage heart surgery had a protective effect (28% in operated patients vs. 60% in non-operated when it was indicated). Mortality was significantly higher in stages B (52.4%) and C (52.9%) and the prognosis was better for patients in stage B who underwent surgery immediately (mortality 50%) compared to those where surgery was delayed (58%) or not performed (74%). Only one patient in stage C underwent surgery. CONCLUSIONS: Patients with liver cirrhosis and infective endocarditis have a poorer prognosis only in stages B and C. Early surgery must be performed in stages A and although in selected patients in stage B when indicated.

Docosahexaenoic acid promotes micron scale liquid-ordered domains. A comparison study of docosahexaenoic versus oleic acid containing phosphatidylcholine in raft-like mixtures.

The understanding of the functional role of the lipid diversity in biological membranes is a major challenge. Lipid models have been developed to address this issue by using lipid mixtures generating liquid-ordered (Lo)/liquid-disordered (Ld) immiscibility. The present study examined mixtures comprising Egg sphingomyelin (SM), cholesterol (chol) and phosphatidylcholine (PC) either containing docosahexaenoic (PDPC) or oleic acid (POPC). The mixtures were examined in terms of their capability to induce phase separation at the micron- and nano-scales. Fluorescence microscopy, electron spin resonance (ESR), X-ray diffraction (XRD) and calorimetry methods were used to analyze the lateral organization of the mixtures. Fluorescence microscopy of giant vesicles could show that the temperature of the micron-scale Lo/Ld miscibility is higher for PDPC than for POPC ternary mixtures. At 37°C, no micron-scale Lo/Ld phase separation could be identified in the POPC containing mixtures while it was evident for PDPC. In contrast, a phase separation was distinguished for both PC mixtures by ESR and XRD, indicative that PDPC and POPC mixtures differed in micron vs nano domain organization. Compared to POPC, the higher line tension of the Lo domains observed in PDPC mixtures is assumed to result from the higher difference in Lo/Ld order parameter rather than hydrophobic mismatch.

Antibiotic susceptibility of different lactic acid bacteria strains.

Five lactic acid bacteria (LAB) strains belonging to species Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis and Streptococcus thermophilus were tested for their susceptibility to 27 antibiotics. The minimum inhibitory concentrations of each antimicrobial were determined using a microdilution test. Among the strains a high susceptibility was detected for most of the cell-wall synthesis inhibitors (penicillins, cefoxitin and vancomycin) and resistance toward inhibitors of DNA synthesis (trimethoprim/sulfonamides and fluoroquinolones). Generally, the Lactobacillus strains were inhibited by antibiotics such as chloramphenicol, erythromycin and tetracycline at breakpoint levels lower or equal to the levels defined by the European Food Safety Authority. Despite the very similar profile of S. thermophilus LC201 to lactobacilli, the detection of resistance toward erythromycin necessitates the performance of additional tests in order to prove the absence of transferable resistance genes.

Surgical treatment for infective endocarditis in elderly patients.

OBJECTIVES: We evaluate the clinical, echographic and prognostic characteristics of infective endocarditis (IE) in a large population of elderly patients, and the results of surgical approach. METHODS: Multicentric, prospective, observational cohort study with 961 consecutive left-sided IE: 356 patients aged ≥65 years were compared with 605 younger. Indications for cardiac surgery, potential surgical risk, time and outcome, were compared. RESULTS: Hospital-acquired endocarditis, comorbidity, renal failure and septic shock were more frequent in elderly, but embolisms were less. Intracardiac destruction and ventricular failure were similar in both groups, but significantly fewer elderly patients underwent cardiac surgery (36% vs 51%; p < 0.01), and this group showed a worse outcome (43.2% of mortality vs 27% in younger; p < 0.01), resulting age as an independent predictor of mortality (OR: 1.02 CI95%: 1.01-1.03). Compared with medical treatment, surgery showed lower percentages of mortality compared with medical treatment (23.3% vs 31.3%; p = 0.03) in younger group, but a high mortality was observed with both procedures (47.6% vs 40.3%; p = 0.1) in the elderly. CONCLUSIONS: Although similar percentages of heart failure and intracardiac complications, increasing age is associated with higher mortality in IE. Lower rates of surgical treatment and a worse outcome after operation are common features in elderly patients.

Streptococcus agalactiae left-sided infective endocarditis. Analysis of 27 cases from a multicentric cohort.

SUMMARY OBJECTIVE: To evaluate the current trends in the clinical characteristics and the prognosis of Streptococcus agalactiae infective endocarditis (IE), uncommon disease associated with high mortality. METHODS: Descriptive analysis of 27 cases of a large cohort (961 episodes) of infective endocarditis collected in seven hospitals of Andalusia (Spain) between 1984 and 2008. RESULTS: Native valves were affected in most cases (85. 2%), multiple valves were frequently involved (22.2%). The median age of the patients was 65 (51-76) years (59.3% men), with a comorbidity, according to the Charlson index, of 2.6+/-2.3. The most frequent underlying diseases were diabetes mellitus (25.9%), chronic obstructive pulmonary disease (14.8%), neoplasms (14.8%), urological disorders (11%) and chronic liver disease (11%). Clinical presentation was characterized by rapid worsening (median of 9 (5.7-15) days from onset of symptoms until diagnosis), a high rate of embolisms (37%) and cardiac complications (abscesses, fistulas or valve rupture) - 37% of cases. Surgery was performed in 12 patients (44.4%) and a high mortality (40.7%) was observed. CONCLUSION: S. agalactiae IE is a serious disease with aggressive course and high mortality rate and affects patients with debilitating diseases. We must be alert of the development of complications and consider early valve surgery when it is necessary.

In vitro inhibition of fungal activity by macrophage-mediated sequestration and release of encapsulated amphotericin B nanosupension in red blood cells.

The efficacy of antifungal treatment has been diminished by the biodistribution limitations of amphotericin B (AmB) due to its pharmacological profile, as well as the severe side effects it causes. A cellular drug-delivery system, which incorporates human erythrocytes (RBCs) loaded with an AmB nanosuspension (AmB-NS), is developed in order to improve antifungal treatment. AmB-NS encapsulation in RBCs is achieved by using hypotonic hemolysis, leading to intracellular AmB amounts of 3.81 +/- 0.47 pg RBC(-1) and an entrapment efficacy of 15-18%. Upon phagocytosis of AmB-NS-RBCs, leukocytes show a slow AmB release over ten days, and no alteration in cell viability. This results in an immediate, permanent inhibition of intra- and extracellular fungal activity. AmB-NS-RBC-leukocyte-mediated delivery of AmB is efficient in amounts 1000 times lower than the toxic dose. This drug-delivery method is effective for the transport of water-insoluble substances, such as AmB, and this warrants consideration for further testing.

[Application of activated recombinant factor VII (rFVIIa, NovoSeven) in neonatology].

Recombinant activated factor VII is vitamin K dependent glycoprotein, similar as the same in the human blood plasma. First has been described in application for hemophilia, where are presented inhibitors of factors VIII and IX, later was proved that it has effectiveness in hemorrhage with different etiology. Clinical experience with this medicine in neonatology is still small, but in the latest years there is some data about its effectiveness in hemorrhage in term and preterm newborns. The data shows that there is low risk of side effects of the recombinant activated factor VII in neonatal period. But still has been described as a experimental treatment.

Do bacteria have an electric permanent dipole moment?

In the scientific literature in the last 40 years, some data for the permanent dipole moment and the electric polarizability of Escherichia coli can be found [S.P. Stoylov, Colloid Electro-Optics - Theory, Techniques and Application, Academic Press, London, 1991]. In this paper the data based mainly on electro-optic investigation is considered as much as some dipolophoretic (most often called dielectrophoretic) studies. Serious grounds are found to doubt the conclusions made for the electric dipole moments of bacteria by one of the authors of this paper (SPS) and by some other researchers. This concerns both the permanent dipole moment and the electric charge dependent polarizabilities of E. coli. Here, along with the discussion of the old experimental data, new experimental data are shown for a strain of E. coli HB101. The conclusions from the analysis of the old and the new experimental data is that they do not provide correct evidence for the presence of a permanent dipole moment. It seems that all statements for the existence of electric permanent dipole moment in bacteria [S.P. Stoylov, Colloid Electro-Optics - Theory, Techniques and Application, Academic Press, London, 1991; S.P. Stoylov, S. Sokerov, I. Petkanchin, N. Ibroshev, Dokl. AN URSS 180 (1968) 1165; N.A. Tolstoy, A.A. Spartakov, A.A. Trusov, S.A. Schelkunova, Biofizika 11 (1966) 453; V. Morris, B. Jennings, J. Chem. Soc. Faraday Trans. II 71 (1975) 1948; V. Morris, B. Jennings, J. Colloid Interface Sci. 55 (1978) 313; S.P. Stoylov, V.N. Shilov, S.S. Dukhin, S. Sokerov, I. Petkanchin, in: S.S. Dukhin (Ed.), Electro-optics of Colloids, Naukova Dumka, Kiev, 1977 (in Russian).] based on electro-optic studies are result of incorrect interpretation. Therefore, they should be further ignored.

[Biophysical models in investigations of exogenous surfactant activities on the surface tension and their theurapeutic effectiveness].

Surfactant therapy leads to significant clinical improvement in infants at risk for, or having, respiratory distress syndrome (RDS). The development of exogenous surfactant (ES) as a therapy for neonatal respiratory disorders was a significant advance in neonatal intensive care that has led to a decrease in neonatal mortality. Two broad categories of surfactants are available for exogenous therapy: surfactants derived from animal sources or 'natural' surfactants; and synthetic surfactants. The physical properties of natural and synthetic surfactants have been studied using techniques such as the Wilhelmy surface balance and the bilayer black film (BBF) method. Here we report some data from a comparative study of ES (Exosurf, Survanta, Curosurf and Alveofact) and clinical samples of tracheal aspirate (TA) of newborns with RDS treated with Curosurf. Measured interfacial physico-chemical parameters prove "better" properties in vitro of the surfactant proteins (SP-B and SP-C) containing preparations Curosurf and Alveofact. Their properties are similar, Alveofact showing a higher surface tension lowering capacity under dynamic conditions. A compendious comparison of results for dynamic surface properties of monolayers of TA from newborns treated with Curosurf with data for newborns treated with Exosurf is presented. Both ES yield the desired lowering of the surface tension during cyclic film compression, being larger after treatment with Curosurf. Observations concerning the properties of BFF of ES (dependence on surfactant concentration, adsorption time, film drainage time and BFF formation time) are also reported and discussed.

Magnetite-loaded carrier erythrocytes as contrast agents for magnetic resonance imaging.

Superparamagnetic iron oxide nanoparticles (SPIONs) are commonly used in magnetic resonance imaging (MRI), but their fast phagocytosis makes them less than ideal for this application. To circumvent the lymphocyte-macrophage system, we encapsulated SPIONs into red blood cells (RBCs). For loading, the RBC's membrane was opened by swelling under hypoosmotic conditions and subsequently resealed. In this work, we demonstrate that SPIONs can be loaded into RBCs in a concentration sufficient to obtain strong contrast enhancement in MRI.

Clinical markers in adult offspring of families with and without Balkan Endemic Nephropathy.

Balkan Endemic Nephropathy (BEN) is a kidney disease that progresses slowly. Only a few studies have investigated renal clinical markers in offspring of BEN families before the onset of the disease. This project aimed to determine whether kidney function and structure are altered in BEN offspring compared with non-BEN offspring. The study population consisted of 102 adult BEN offspring and a control group of 99 non-BEN offspring. We collected urine and blood samples, and conducted face-to-face interviews, physical examinations and ultrasound measurements of the kidney. Total protein, albumin, beta2-microglobulin and creatinine in urine, creatinine and urea in serum, and creatinine clearance (CCR) were determined. Two risk factors were assessed: first, the overall status of being an offspring from a BEN family, and second, the specific status of a mother and/or father with BEN. The data were analyzed using linear regression. After adjusting for confounders, we found that kidney length and minimal cortex width in BEN offspring were significantly decreased. Urine concentrations of total protein, albumin, and beta2-microglobulin were higher in BEN offspring. Regarding parental history, the associations were statistically significant only for the offspring of mothers who had BEN, with the exception of minimal cortex width, which showed no parental difference. For CCR, we did not identify a statistically significant effect for BEN offspring status nor for parental history. In conclusion, adult offspring of BEN families can be characterized by shorter kidney length and an increased excretion of albumin, total protein, and beta2-microglobulin, in particular, when the mother had BEN.

Expression of alpha5beta1 integrin and fibronectin during early pregnancy in pigs.

The implantation in pig is superficial and non-invasive, involving phases of apposition, adhesion and attachment of conceptuses to endometrial surface epithelium. The role of integrins and ECM proteins is suggested. In the study, the expression of beta5beta1 integrin and FN on conceptus trophectoderm and endometrium during implantation and early pregnancy was investigated. The immunohistochemical localization of alpha5beta1 integrin and FN was performed on formalin-fixed, paraffin-embedded tissue sections using the ABC method. The results indicate that both conceptus and uterus expressed alpha5beta1 integrin and FN during early porcine pregnancy. The most intensive staining for alpha5beta1 integrin and FN was found in conceptus trophectoderm and endometrial surface epithelium in all investigated periods. During placentation the immunohistochemical staining for both alpha5beta1 integrin and FN was increased in trophectoderm and all endometrial structures. Since placenta in pigs is non-invasive, it can be suggested that both alpha5beta1 integrin and FN participate in molecular events leading to successful implantation and placentation in species with true epitheliochorial placenta.

Composite lipid polyelectrolyte capsules templated on red blood cells: fabrication and structural characterisation.

Charged phospholipids and mixtures of charged phospholipids with zwitterionic lipids were adsorbed onto polyelectrolyte capsules templated on erythrocytes. The assembly was proved by means of electrophoretic mobility measurements, confocal laser scanning microscopy and flow cytometry. Freeze-fracture electron microscopy proved that the phospholipids assemble as bilayers or multilayers. Single particle light scattering showed that bilayers composed of anionic lipids can be intercalated between subsequent polyelectrolyte inter-layers in a regular manner. Neutral lipids can form multilayers. A pronounced decrease in capsule permeability for small polar dyes upon lipid adsorption was followed by confocal laser scanning microscopy.

Health risk for children from lead and cadmium near a non-ferrous smelter in Bulgaria.

In an interdisciplinary approach the exposure to lead and cadmium of the population living in the vicinity of a Bulgarian non-ferrous metallurgical plant was studied to determine the routes of exposure. Two exposed villages situated 4 and 6 km SW and W, respectively,--in the plume of the plant's emissions--and a village for comparison situated 14 km SE of the plant were selected for the study. The blood of children aged 3-13 years from the exposed villages (n = 111) and the village for comparison (n = 18) was analyzed for lead (PbB) and cadmium (CdB) as well as for free erythrocyte protoporphyrin (FEP) and haematological parameters. A personal questionnaire concerning nutrition habits and life-style was filled in by interviewers of the children's parents. The cadmium levels in blood were within the variation range stated for Bulgaria. The exposed children had a mean CdB of 0.38 +/- 0.18 microgram/l (range 0.10-0.90 microgram/l) and the comparison group 0.31 +/- 0.35 microgram/l (0.06-1.42 micrograms/l). The mean PbB value measured in the exposed children was 240 +/- 96 micrograms/l (77-631 micrograms/l), while the mean value of PbB for the reference group was 149 +/- 57 micrograms/l (63-285 micrograms/l). Individual PbB levels > 150 and > 200 micrograms/l were found in 85% and 62%, respectively, of the exposed children and FEP > 80 micrograms/dl erythrocytes was detected in 21%. There was a concentration-response and a concentration-effect relationship (r = 0.51, D > 0.001) between PbB and FEP. According to the questionnaire only 10.3% of the interviewed families in the two exposed villages buy all food from the market, the others produce a significant part of their food themselves. The PbB levels of children who consume certain foods of local production (cabbage, cereals, poultry, and beef) were significantly higher compared to the PbB values of children whose families purchase these foods from the market. No significant relationship between lead in children's blood and the use of tin and ceramic utensils, drinking and irrigation water, hygiene habits, family tobacco smoking habits, playgrounds and traffic intensity were revealed. There was a significantly higher incidence of prematurely born babies in the exposed population. The proven relationship between lead intake via the food chain and existing significant health risks is the basis for risk communication. Measures aimed at healthier nutrition and life-style intend to contribute to decreasing the risk of the population living in the smelter region.

Permeation of macromolecules into polyelectrolyte microcapsules.

Polyelectrolyte microcapsules (PEMCs) have been prepared by coating red blood cells with the polyelectrolytes poly(styrenesulfonate), poly(allylamine hydrochloride), and dextran sulfate applying the layer-by-layer technique with subsequent dissolution of the core. The capsule permeability for human serum albumin (HSA) was studied as a function of the ionic strength and pH by means of confocal microscopy. PEMCs produced with dextran sulfate and poly(allylamine hydrochloride) show a significant increase in permeability for HSA at salt concentrations over 1 mM. For PEMCs prepared with poly(styrenesulfonate) and poly(allylamine hydrochloride) the limiting salt concentration is 5 mM. No pH dependence for permeation was observed. A correlation between the permeation and adsorption of HSA on the PEMC walls was investigated. Finally, a mechanism for the permeability, combining electrostatic interactions, and the presence of pores in the polymer layers is presented confirmed by the considerable increase of permeation of charged molecules in the presence of salt and the permeation of neutral molecules regardless of the ionic strength.

Electrophoretic and aggregation behavior of bovine, horse and human red blood cells in plasma and in polymer solutions.

The electrophoretic mobility of native and glutaraldehyde-fixed bovine, human, and horse red blood cells (RBC) was investigated as a function of ionic strength (5-150 mM) and concentration of 464 kDa dextran (2 and 3 g/dl); RBC aggregation in autologous plasma and in dextran solutions was also measured. In agreement with previous observations, human and horse RBC form stable rouleaux whereas bovine RBC do not aggregate in either plasma or in dextran 464 kDa solutions. Electrophoretic measurements showed a species-dependent adsorption and depletion of dextran that can be theoretically evaluated. Adsorption of polymer is not a prerequisite for RBC aggregation (bovine RBC show the highest amount of adsorbed dextran yet do not aggregate). Aggregate formation thus occurs as long as the Gibbs free energy difference, given by the osmotic pressure difference between the bulk phase and the polymer-depleted region between two RBC, is larger than the steric and electrostatic repulsive energy contributed by the macromolecules present on the RBC surface. With increasing bulk-phase polymer concentration the depletion layer thickness decreases and the amount of adsorbed macromolecules increases, thereby resulting in an increase of the repulsive component of the interaction energy and decreased aggregation. We thus view electrophoretic measurements of RBC in various media as an important tool for understanding polymer behavior near the red cell surface and hence the mechanisms involved in RBC aggregation.