Bioinspired green synthesis of ZnO nanoparticles by marine-derived Streptomyces plicatus and its multifaceted biomedicinal properties.

The present study explores the bioinspired green synthesis of zinc oxide nanoparticles (ZnONPs) using marine Streptomyces plicatus and its potent antibacterial, antibiofilm activity against dental caries forming Streptococcus mutans MTCC and S. mutans clinical isolate (CI), cytotoxicity against oral KB cancer cells, hemolysis against blood erythrocytes and artemia toxicity. The bioinspired ZnONPs showed a distinctive absorption peak at 375 nm in UV-Vis spectra, the FT-IR spectra divulged the active functional groups, and XRD confirmed the crystalline nature of the nanoparticles with an average grain size of 41.76 nm. SEM analysis evidenced hexagonal morphology, and EDX spectra affirmed the presence of zinc. The ZnONPs exerted higher antagonistic activity against S. mutans MTCC (Inhibitory zone: 19 mm; MIC: 75 µg/ml) than S. mutans CI (Inhibitory zone: 17 mm; MIC: 100 µg/ml). Results of biofilm inhibitory activity showed a concentration-dependent reduction with S. mutans MTCC (15 %-95 %) more sensitive than S. mutans CI (13 %-89 %). The 50 % biofilm inhibitory concentration (BIC50) of ZnONPs against S. mutans MTCC was considerably lower (71.76 µg/ml) than S. mutans CI (78.13 µg/ml). Confocal Laser Scanning Microscopic visuals clearly implied that ZnONPs effectively distorted the biofilm architecture of both S. mutans MTCC and S. mutans CI. This was further bolstered by a remarkable rise in protein leakage (19 %-85 %; 15 %-77 %) and a fall in exopolysaccharide production (34 mg-7 mg; 49 mg-12 mg). MTT cytotoxicity of ZnONPs recorded an IC50 value of 22.06 µg/ml against KB cells. Acridine orange/ethidium bromide staining showed an increasing incidence of apoptosis in KB cells. Brine shrimp cytotoxicity using Artemia salina larvae recorded an LC50 value of 78.41 µg/ml. Hemolysis assay substantiated the biocompatibility of the ZnONPs. This study underscores the multifaceted application of bioinspired ZnONPs in dentistry.

Synthesis, theoretical studies, antibacterial, and antibiofilm activities of novel azo-azomethine chelates against the pathogenic bacterium Proteus mirabilis.

2-((1-(4-((2,4,6-trioxohexahydropyrimidin-5-yl)diazenyl) phenyl) ethylidene) amino) benzoic acid (H3L), and its V(IV), Co(II), Ni(II), Cu(II), Pd(II) and Ag(I) chelates were synthesized. They were defined using multiple spectral and analytical techniques. With the exception of Ag(I) chelate, all chelates possessed non-electrolytic character. Square pyramidal shape was proposed for V(IV) chelate and Square planar for the other chelates. The analysis of functional group bands of H3L and its coordination compounds alludes that H3L chelated as neutral tetradentate via nitrogen atoms of azo and azomethine groups, oxygen atom of carbonyl of barbituric acid and OH of the carboxylic group. TG/DTG predicted the thermal behaviors of all compounds. The antibacterial activity of H3L and its coordination compounds was conducted against Proteus mirabilis at concentrations of 250, 500, and 1000 µg/mL. Ag(I) at 1000 µg/mL, showed the most inhibiting potency against P. mirabilis and registered zone of inhibition of 28.33 ± 0.84 mm and highest biofilm inhibition of 70.31%. At 50 Gy of gamma irradiation, the reducing effect of Ag(I) chelate was improved. The protein interruption of P. mirabilis was greatly interrupted by increasing the concentration of the chaletes. Also, Ag(I) showed the highest cytotoxicity with IC50 value of 11.5 µg/ mL. The novelty of this study is the synthesis of a new azo-Schiff base and this is almost the first publication of the effect of azo-Schiff ligands against that bacterial strain P. mirabilis.

Role of endothelial hyaluronan in peritoneal membrane transport and disease conditions during peritoneal dialysis.

Peritoneal membrane dysfunction in peritoneal dialysis (PD) is primarily attributed to angiogenesis; however, the integrity of vascular endothelial cells can affect peritoneal permeability. Hyaluronan, a component of the endothelial glycocalyx, is reportedly involved in preventing proteinuria in the normal glomerulus. One hypothesis suggests that development of encapsulating peritoneal sclerosis (EPS) is triggered by protein leakage due to vascular endothelial injury. We therefore investigated the effect of hyaluronan in the glycocalyx on peritoneal permeability and disease conditions. After hyaluronidase-mediated degradation of hyaluronan on the endothelial cells of mice, macromolecules, including albumin and ß2 microglobulin, leaked into the dialysate. However, peritoneal transport of small solute molecules was not affected. Pathologically, hyaluronan expression was diminished; however, expression of vascular endothelial cadherin and heparan sulfate, a core protein of the glycocalyx, was preserved. Hyaluronan expression on endothelial cells was studied using 254 human peritoneal membrane samples. Hyaluronan expression decreased in patients undergoing long-term PD treatment and EPS patients treated with conventional solutions. Furthermore, the extent of hyaluronan loss correlated with the severity of vasculopathy. Hyaluronan on endothelial cells is involved in the peritoneal transport of macromolecules. Treatment strategies that preserve hyaluronan in the glycocalyx could prevent the leakage of macromolecules and subsequent related complications.

Ultrastructural Alterations of the Glomerular Filtration Barrier in Fish Experimentally Exposed to Perfluorooctanoic Acid.

Per- and polyfluoroalkyl substances can be referred to as the most critical group of contaminants of emerging concern. They can accumulate in high concentration in the kidney and are known to potentially affect its function. Nonetheless, there is a lack of knowledge about their morphopathological effect on the glomerular filtration barrier. Since previous research suggests perfluorooctanoic acid (PFOA) induces glomerular protein leakage, the glomerular filtration barrier of 30 carp from the same parental stock (10 unexposed; 10 exposed to 200 ng L-1 of PFOA; and 10 exposed to 2 mg L-1 of PFOA for 56 days) was screened for possible PFOA-induced ultrastructural lesions in order to shed light on the related pathophysiology. PFOA exposure affected the glomerular filtration barrier in carp experimentally exposed to 2 mg L-1, showing ultrastructural alterations compatible with glomerulonephrosis: podocyte effacement, reduction of filtration slits and filtration slit diaphragms, basement membrane disarrangement, and occurrence of proteinaceous material in the urinary space. The results of the present research confirm the glomerular origin of the PFOA-induced protein leakage and can contribute to the mechanistic comprehension of PFOA's impact on renal function and to the assessment of the exposure effect of environmental pollutants on animals and humans, according to the One Health approach.

Radiation synthesis and in vitro evaluation of the antimicrobial property of functionalized nanopolymer-based poly (propargyl alcohol) against multidrug-resistance microbes.

Pathogenic microorganisms are responsible for many diseases in biological organisms, including humans. Many of these infections thrive in hospitals, where people are treated with medicines and certain bacteria resist those treatments. Consequently, this research article aims to develop efficient antimicrobial material-based conjugated and functionalized polypropargyl alcohol nanoparticles (nano-PGA) synthesized by gamma irradiation. The monomer of PGA was polymerized in various mediums (water (W), chloroform (Ch), and dimethylformamide (DMF)) without catalysts under the action of γ-rays, producing π-conjugated and colored functional nano-PGA polymers. Nano-PGA is a versatile polymer demonstrated here as suitable for creating next-generation of antimicrobial systems capable of effectively preventing and killing various pathogenic microorganisms. The novelty here is the development of polymeric nanostructures by changing the solvent and irradiation doses. The antimicrobial property of nano-PGA (nanostare-like antibody structure) was examined against different pathogenic bacteria and unicellular fungi. Nano-PGA-DMF exhibits significant antimicrobial potential against Staphylococcus aureus (S. aureus) (20.20 mm; zone of inhibition (ZOI), and 0.47 µg/mL; minimum inhibitory concentration (MIC), followed by Escherichia coli (E. coli) (14.50 mm; ZOI, and 1.87 µg/mL; MIC, and Candida albicans (C.albicans) (12.50 mm; ZOI, and 1.87 µg/mL; MIC). In antibiofilm results, the highest inhibition percentage of the synthesized nano-PGA-W, nano-PGA-Ch, and nano-PGA-DMF was documented for S. aureus (17.01%, 37.57%, and 80.27%), followed by E. coli (25.68%, 55.16% and 78.11%), and C.albicans (40.10%, 62.65%, and 76.19%), respectively. The amount of bacterial protein removed is directly proportional after increasing the concentration of nano-PGA-W, nano-PGA-Ch, and nano-PGA-DMF samples (at different concentrations) and counted to be 70.58, 102.89, and 200.87 µg/mL, respectively following the treatment with 1.0 mg/mL of each sample. It was found that the nano-PGA polymer prepared in DMF has better antimicrobial activity than one prepared in chloroform than in water.

Sealing ability of three different root repair materials for furcation perforation repair: An in vitro study.

AIM: The present study aimed to evaluate and compare the sealing ability of mineral trioxide aggregate (MTA)-Angelus, Biodentine™, and EndoSequence cement in furcation perforations using protein leakage assessment. MATERIALS AND METHODS: The present study was conducted using seventy extracted human maxillary and mandibular molars with intact furcation. The samples were randomly allocated into three groups (n = 20) based on repair material used: Group 1 - MTA-Angelus, Group 2 - Biodentine™, and Group 3 - EndoSequence. Two additional groups served as positive and negative controls (n = 5). Using the leakage assessment apparatus, the presence of protein was detected with a reagent (Coomassive Brilliant Blue) every day for 60 days. One-way ANOVA and post hoc Tukey's test were used for statistical analysis using SPSS software. RESULTS: All the groups exhibited protein leakage from day 1. Biodentine showed minimum (0.1201 mg/ml), MTA showed maximum (0.3738 mg/ml), and EndoSequence had intermediate (0.2465 mg/ml) leakage. None in the negative control and all of the positive control specimens leaked during the experimental period of 60 days. CONCLUSION: The newer biomaterials, Biodentine and Endosequence with better handling properties, could be used as alternatives to MTA-Angelus while repairing furcation perforations.

Enhanced coagulation by high-frequency ultrasound in Microcystis aeruginosa-laden water: Strategies and mechanisms.

Ultrasonic treatment has attracted much attention because of its physical and chemical effects that are distinct from those of chemical agents. In particularly, high-frequency ultrasound is known as an effective method because the theoretical resonance frequency of the gas vesicles in Microcystis aeruginosa is in the high frequency range (>100 kHz), which causes gas vesicles collapse and changes the settleability of the algal cells. In this work, the effects of the ultrasonic frequency, acoustic power density and duration on enhancing coagulation to remove turbidity in algae-laden water were studied. In order to explain the mechanism, the morphology of algae cells, the changes in extracellular organic substances, the zeta potential and the formation of hydroxyl radicals were analyzed systematically. Finally, Zeta potentials and flocs morphology after adding PAC were investigated to verify the mechanism. The results showed that the frequency exhibited fewer effects than power and duration on coagulation. SEM images showed that there were more severe cellular damages at 430 and 740 kHz than other frequencies. Sonication could cause the collapse of gas vesicle inside the cell, which was due to the instantaneous high pressure generated by the ultrasonic cavitation instead of the resonance. Furthermore, sonication would result in an increase in proteins in extracellular organic matter (EOM) with continuous ultrasonic irradiation, indicating that a small amount of proteins could promote coagulation and that the accumulation of proteins would inhibit coagulation. Free radical content testing showed that the production of excessive free radicals was often accompanied by a deterioration of the coagulation. The proper mechanical effects were the main mechanism of ultrasonic enhanced coagulation. Thus, it was recommended that the appropriate ultrasonic condition was the one that resulted in a small amount of protein leakage and little generation of free radicals, which occurred at 740 kHz and 0.02 W/mL in approximately 5 min, and would significantly enhance the turbidity removal rate in algae-containing water from approximately 80-90%.

A myopic perspective on the future of protein diagnostics.

Plasma proteome analyses of the future promise invaluable insights into states of health, not only by measuring proteins whose role it is to ensure blood homeostasis, but increasingly also as a window into the health of practically any tissue in the body via so-called leakage protein biomarkers. Realizing more of this vast potential will require progress along many lines. Here we discuss the main ones, such as optimal selection of target proteins, affinity reagents, immunoassay formats, samples, and applications, with a view from ongoing work in our laboratory.

Enhanced antibacterial effects of green synthesized ZnO NPs using Aristolochia indica against Multi-drug resistant bacterial pathogens from Diabetic Foot Ulcer.

BACKGROUND: Increased incidence of Multi-drug resistance in microorganisms has become the greatest challenge in the treatment of Diabetic Foot Ulcer (DFU) and urges the need of a new antimicrobial agent. In this study, we determined the bactericidal effects of ZnO nanoparticles (ZnO NPs) green synthesized from Aristolochia indica against Multi-drug Resistant Organisms (MDROs) isolated from pus samples of DFU patients attending in a tertiary care hospital in South India. METHODS: ZnO NPs were characterized by UV-vis-DRS spectroscopy, Atomic Force Microscopy (AFM), Transmission Electron Microscopy (TEM) and for its zeta potential value. MIC/MBC assays were performed to determine bactericidal or bacteriostatic effects. Time-kill assays, Protein leakage and Flow cytometric analysis evaluated bacterial cell death at 1x MIC and 2x MIC concentrations of ZnO NPs. RESULTS: ZnO NPs of size 22.5nm with a zeta potential of -21.9±1mV exhibited remarkable bactericidal activity with MIC/MBC ranging from 25 to 400µg/ml with a significant reduction in viable count from 2h onwards. Protein leakage and Flow cytometric analysis confirmed bacterial cell death due to ZnO NPs. CONCLUSION: This study concluded that green synthesis protocol offers reliable, eco-friendly approach towards the development of antimicrobial ZnO NPs to combat antibiotic drug resistance.

Multipurpose efficacy of ZnO nanoparticles coated by the crustacean immune molecule ß-1, 3-glucan binding protein: Toxicity on HepG2 liver cancer cells and bacterial pathogens.

The effective treatment of cancer and bacterial pathogens are two key challenges in modern nanomedicine. Here, zinc oxide nanoparticles (ZnO NPs) were fabricated using the crustacean immune molecule ß-1, 3- glucan binding protein (Phß-GBP, 100kDa) purified from the heamolymph of Paratelphusa hydrodromus. ß-GBP coated zinc oxide nanoparticles (Phß-GBP-ZnO NPs) were characterized by UV-vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and high resolution-transmission electron microscopy (HR-TEM) analyses. Phß-GBP-ZnO NPs inhibited the growth of Staphylococcus aureus and Proteus vulgaris. Protein and nucleic acid leakage assays showed that Phß-GBP-ZnO NPs facilitate membrane permeability leading to cell death. The antibacterial activity of Phß-GBP-ZnO NPs was due to the high level of reactive oxygen species (ROS) release from bacterial cells post-treatment with 75µg/mL of Phß-GBP-ZnO NPs. Confocal laser scanning microscopy pointed out that biofilm thickness was highly reduced post-treatment with nanoparticles. Cytotoxicity on human liver carcinoma (HepG2) cells highlighted that 75µg/mL of Phß-GBP-ZnO NPs inhibited viability of HepG2 cells. Phase contrast microscopy showed key morphological changes of HepG2 cells post-treatment with Phß-GBP-ZnO NPs. Overall, Phß-GBP-ZnO NPs can be further considered for the development of novel drugs against microbial pathogens and HepG2 cells.

Time-kill kinetics and biocidal effect of Euclea crispa leaf extracts against microbial membrane.

OBJECTIVE: To evaluate antimicrobial potential of the fractions partitioned from Euclea crispa leaf extract and determination of their impact on cell membrane disruption. METHODS: Antimicrobial potentials were evaluated via susceptibility test, determination of minimum inhibitory concentrations (MICs) and time-kill kinetics of the potent fractions. Degree of membrane disruption was determined by the amount of proteins and nucleotides released from within the cells and SEM images of the membrane after 120 min of treatment. RESULTS: The largest inhibition zone (25.5 ± 0.50 mm) was obtained by ethylacetate fraction against Aeromonas hydrophilla at 10 mg/mL. The lowest MIC (0.16 mg/mL) was exhibited by n-butanol and ethylacetate fractions against test bacteria while all fractions exhibited MIC values between 0.31 and 1.25 mg/mL against susceptible yeast. n-Butanol fraction achieved absolute mortality against Bacillus pumulis (B. pumulis) and Klebsiella pneumoniae (K. pneumoniae) after 90 and 120 min contact time respectively at 1 × MIC. Total mortality also achieved by n-hexane fraction against B. pumulis and K. pneumoniae after 90 and 120 min respectively at 2 × MIC. Ethylacetate fraction achieved absolute mortality against both bacteria after 120 min at 2 × MIC. n-Hexane fraction achieved total mortality against Candida albicans after 120 min at 1 × MIC. Maximum amount of proteins (0.566 µg/mL) was released from K. pneumoniae by n-butanol fraction at 2 × MIC after 120 min of treatment while the maximum amount of nucleotides released (4.575 µg) was from B. pumulis by n-hexane fraction under similar condition. CONCLUSION: This study suggests the leaf of Euclea crispa a source of bioactive compound with membrane attack as one of the mechanisms of its biocidal action.

Time-kill kinetics and biocidal effect of Euclea crispa leaf extracts against microbial membrane

OBJECTIVE@#To evaluate antimicrobial potential of the fractions partitioned from Euclea crispa leaf extract and determination of their impact on cell membrane disruption.@*METHODS@#Antimicrobial potentials were evaluated via susceptibility test, determination of minimum inhibitory concentrations (MICs) and time-kill kinetics of the potent fractions. Degree of membrane disruption was determined by the amount of proteins and nucleotides released from within the cells and SEM images of the membrane after 120 min of treatment.@*RESULTS@#The largest inhibition zone (25.5 ± 0.50 mm) was obtained by ethylacetate fraction against Aeromonas hydrophilla at 10 mg/mL. The lowest MIC (0.16 mg/mL) was exhibited by n-butanol and ethylacetate fractions against test bacteria while all fractions exhibited MIC values between 0.31 and 1.25 mg/mL against susceptible yeast. n-Butanol fraction achieved absolute mortality against Bacillus pumulis (B. pumulis) and Klebsiella pneumoniae (K. pneumoniae) after 90 and 120 min contact time respectively at 1 × MIC. Total mortality also achieved by n-hexane fraction against B. pumulis and K. pneumoniae after 90 and 120 min respectively at 2 × MIC. Ethylacetate fraction achieved absolute mortality against both bacteria after 120 min at 2 × MIC. n-Hexane fraction achieved total mortality against Candida albicans after 120 min at 1 × MIC. Maximum amount of proteins (0.566 μg/mL) was released from K. pneumoniae by n-butanol fraction at 2 × MIC after 120 min of treatment while the maximum amount of nucleotides released (4.575 μg) was from B. pumulis by n-hexane fraction under similar condition.@*CONCLUSION@#This study suggests the leaf of Euclea crispa a source of bioactive compound with membrane attack as one of the mechanisms of its biocidal action.

Time-kill kinetics and biocidal effect of Euclea crispa leaf extracts against microbial membrane

Objective To evaluate antimicrobial potential of the fractions partitioned from Euclea crispa leaf extract and determination of their impact on cell membrane disruption. Methods Antimicrobial potentials were evaluated via susceptibility test, determination of minimum inhibitory concentrations (MICs) and time-kill kinetics of the potent fractions. Degree of membrane disruption was determined by the amount of proteins and nucleotides released from within the cells and SEM images of the membrane after 120 min of treatment. Results The largest inhibition zone (25.5 ± 0.50 mm) was obtained by ethylacetate fraction against Aeromonas hydrophilla at 10 mg/mL. The lowest MIC (0.16 mg/mL) was exhibited by n-butanol and ethylacetate fractions against test bacteria while all fractions exhibited MIC values between 0.31 and 1.25 mg/mL against susceptible yeast. n-Butanol fraction achieved absolute mortality against Bacillus pumulis (B. pumulis) and Klebsiella pneumoniae (K. pneumoniae) after 90 and 120 min contact time respectively at 1 × MIC. Total mortality also achieved by n-hexane fraction against B. pumulis and K. pneumoniae after 90 and 120 min respectively at 2 × MIC. Ethylacetate fraction achieved absolute mortality against both bacteria after 120 min at 2 × MIC. n-Hexane fraction achieved total mortality against Candida albicans after 120 min at 1 × MIC. Maximum amount of proteins (0.566 μg/mL) was released from K. pneumoniae by n-butanol fraction at 2 × MIC after 120 min of treatment while the maximum amount of nucleotides released (4.575 μg) was from B. pumulis by n-hexane fraction under similar condition. Conclusion This study suggests the leaf of Euclea crispa a source of bioactive compound with membrane attack as one of the mechanisms of its biocidal action.

Bactericidal mechanism of electrolyzed oxidizing water against Pseudomonas aeruginosa / 中国感染控制杂志

Objective To investigate the bactericidal mechanism of electrolyzed oxidizing water (EOW) against Pseudomona aeruginosa (P.aeruginosa).Methods Bactericidal mechanism of EOW against P.aeruginosa was studied through intracellular protein leakage,nucleic acid,and cell membrane calcium ion permeability,2 ％ glutaraldehyde was used as positive control group,and normal saline (NS) was used as negative control group.Results The killing rates of EOW and 2％ glutaraldehyde to P.aeruginosa were both＞99.99％ with 30-second contact time,and 100.00％ with 60-second contact time.After 60-second contact with EOW,NS,and 2％ glutaraldehyde,the protein leakage of P.aeruginosa detected by bicinchoninic acid (BCA) were (96.00 ± 7.42),(94.15 ± 7.49),and (216.97 ± 10.35)μg/mL,respectively,difference was significant(F =613.20,P＜0.01),2％ glutaraldehyde group was higher than EOW group and NS group;protein leakage did not change with the increase of contact time(all P＞0.05).Electrophoretogram of random amplified polymorphic DNA showed high intensity dense band between 500-1000 Kb in EOW group and NS group,while 2％ glutaraldehyde group was without amplified bands.The fluorescence intensity of calcium ion of EOW group and 2％ glutaraldehyde group were both lower than that of NS group.Conclusion Bactericidal mechanism of EOW may be due to the damage of membrane permeability of P.aeruginosa,which causes Ca2+ leakage,but fails to cause protein leakage,the damage to nucleic acid is not obvious,DNA may not be a bactericidal target of EOW.

Standardized myrtol attenuates lipopolysaccharide induced acute lung injury in mice.

CONTEXT: Standardized myrtol, an essential oil containing primarily cineole, limonene and α-pinene, has been used for treating nasosinusitis, bronchitis and chronic obstructive pulmonary disease (COPD). OBJECTIVE: To investigate the effects of standardized myrtol in a model of acute lung injury (ALI) induced by lipopolysaccharides (LPS). MATERIALS AND METHODS: Male BALB/c mice were treated with standardized myrtol for 1.5 h prior to exposure of atomized LPS. Six hours after LPS challenge, lung injury was determined by the neutrophil recruitment, cytokine levels and total protein concentration in the bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity in the lung tissue. Additionally, pathological changes and NF-κB activation in the lung were examined by haematoxylin and eosin staining and western blot, respectively. RESULTS: In LPS-challenged mice, standardized myrtol at a dose of 1200 mg/kg significantly inhibited the neutrophile counts (from 820.97 ± 142.44 to 280.42 ± 65.45, 103/mL), protein concentration (from 0.331 ± 0.02 to 0.183 ± 0.01, mg/mL) and inflammatory cytokines level (TNF-α: from 6072.70 ± 748.40 to 2317.70 ± 500.14, ng/mL; IL-6: from 1184.85 ± 143.58 to 509.57 ± 133.03, ng/mL) in BALF. Standardized myrtol also attenuated LPS-induced MPO activity (from 0.82 ± 0.04 to 0.48 ± 0.06, U/g) and pathological changes (lung injury score: from 11.67 ± 0.33 to 7.83 ± 0.79) in the lung. Further study demonstrated that standardized myrtol prevented LPS-induced NF-κB activation in lung tissues. DISCUSSION AND CONCLUSION: Together, these data suggest that standardized myrtol has the potential to protect against LPS-induced airway inflammation in a model of ALI.

Sphingolipids affect fibrinogen-induced caveolar transcytosis and cerebrovascular permeability.

Inflammation-induced vascular endothelial dysfunction can allow plasma proteins to cross the vascular wall, causing edema. Proteins may traverse the vascular wall through two main pathways, the paracellular and transcellular transport pathways. Paracellular transport involves changes in endothelial cell junction proteins, while transcellular transport involves caveolar transcytosis. Since both processes are associated with filamentous actin formation, the two pathways are interconnected. Therefore, it is difficult to differentiate the prevailing role of one or the other pathway during various pathologies causing an increase in vascular permeability. Using a newly developed dual-tracer probing method, we differentiated transcellular from paracellular transport during hyperfibrinogenemia (HFg), an increase in fibrinogen (Fg) content. Roles of cholesterol and sphingolipids in formation of functional caveolae were assessed using a cholesterol chelator, methyl-ß-cyclodextrin, and the de novo sphingolipid synthesis inhibitor myriocin. Fg-induced formation of functional caveolae was defined by association and colocalization of Na+-K+-ATPase and plasmalemmal vesicle-associated protein-1 with use of Förster resonance energy transfer and total internal reflection fluorescence microscopy, respectively. HFg increased permeability of the endothelial cell layer mainly through the transcellular pathway. While MßCD blocked Fg-increased transcellular and paracellular transport, myriocin affected only transcellular transport. Less pial venular leakage of albumin was observed in myriocin-treated HFg mice. HFg induced greater formation of functional caveolae, as indicated by colocalization of Na+-K+-ATPase with plasmalemmal vesicle-associated protein-1 by Förster resonance energy transfer and total internal reflection fluorescence microscopy. Our results suggest that elevated blood levels of Fg alter cerebrovascular permeability mainly by affecting caveolae-mediated transcytosis through modulation of de novo sphingolipid synthesis.

Relationship between membrane damage, leakage of intracellular compounds, and inactivation of Escherichia coli treated by pressurized CO2.

The relationship between membrane damage, leakage of intracellular compounds, and inactivation of Escherichia coli treated by pressurized CO2 was investigated by assessing the inactivation, bacterial cell membrane permeability, the leakage of protein, nucleic acid, and K(+) and Mg(2+) of E. coli. The results indicated that pressurized CO2 treatment induced the leakage of protein in E. coli, but the time of leakage was lagged behind the time of 99% E. coli inactivation, so it was only the secondary phenomenon of inactivation. The inactivation of E. coli was related to the leakage of nucleic acid, K(+) , Mg(2+) induced by the pressurized CO2 treatment. There was direct relationship between the inactivation of E. coli and the damaging effect of pressurized CO2 treatment on the cell membrane of E. coli.

Peritoneal protein leakage, systemic inflammation, and peritonitis risk in patients on peritoneal dialysis.

BACKGROUND: Whether peritoneal protein leakage predicts risk for peritonitis in patients on peritoneal dialysis (PD) is unknown. In this observational cohort study, we aimed to determine that association and, further, to explore if it might be explained by systemic inflammation. ♢ METHODS: We prospectively followed 305 incident PD patients to first-episode peritonitis, censoring, or the end of the study. Demographics, comorbidity score, biochemistry, and peritoneal protein clearance (PrC) were collected at baseline. The predictors of first-episode peritonitis were analyzed prospectively. ♢ RESULTS: During follow-up, 14 868 patient months and 251 episodes of peritonitis were observed. The baseline PrC was 73.2 mL/day (range: 53.2 - 102 mL/day). Patients with a high PrC were prone to be older and malnourished. They also had a higher comorbidity score and higher C-reactive protein values. In 132 first episodes of peritonitis, baseline PrC was shown to be a significant independent predictor after adjustment for age, sex, body mass index, diabetes, residual renal function, hemoglobin, and peritoneal transport rate. Systemic inflammatory markers such as serum albumin, C-reactive protein, and interleukin-6 could not explain the association of PrC and high risk for peritonitis. ♢ CONCLUSIONS: Baseline peritoneal protein leakage was able to independently predict risk for peritonitis, which is not explained by systemic inflammation. The underlying mechanisms should be explored in future.

Studies on the biocidal and cell membrane disruption potentials of stem bark extracts of Afzelia africana (Smith)

We had recently reported antibacterial activity in the crude extract of the stem bark of Afzelia africana (Akinpelu et al., 2008). In this study, we assessed the biocidal and cell membrane disruption potentials of fractions obtained from the crude extract of the plant. The aqueous (AQ) and butanol (BL) fractions exhibited appreciable antibacterial activities against the test bacteria. The minimum inhibitory concentrations of the AQ and BL fractions ranged between 0.313 and 2.5 mg/ml, while their minimum bactericidal concentrations varied between 0.625 and 5.0 mg/ml. Also, the AQ fraction killed about 95.8 percent of E. coli cells within 105 min at a concentration of 5 mg/ml, while about 99.1 percent of Bacillus pumilus cells were killed by this fraction at the same concentration and exposure time. A similar trend was observed for the BL fraction. At a concentration of 5 mg/ml, the butanol fraction leaked 9.8 μg/ml of proteins from E. coli cells within 3 h, while the aqueous fraction leaked 6.5 μg/ml of proteins from the same organisms at the same concentration and exposure time. We propose that the stem bark of Afzelia africana is a potential source of bioactive compounds of importance to the pharmaceutical industry.

Biocidal activity of partially purified fractions from methanolic extract of Garcinia kola (Heckel) seeds on bacterial isolates

The in vitro antibacterial activity of crude methanolic extract of the seeds of Garcinia kola was investigated. The extracts exhibited antibacterial activities with zones of inhibition ranging from 10 mm to 25 mm. Theminimum inhibitory concentration of the diethyl ether fraction was between 0.313 and 5.0 mg/ml, while that of butanol fraction varied from 0.157 to 5.0 mg/ml. The butanol fraction killed about 77% of Bacillus anthracis and 79% of Escherichia coli cells within 120 min at a concentration of 5.0 mg/ml. Protein leakage from the B. anthracis and E. coli cells when exposed to the butanol and diethyl ether fractions was observed. We conclude that Garcinia kola seed extract has a broad spectrum antibacterial activity, with the butanol and diethyl ether fractions being bactericidal as exemplified by the killing rate and protein leakage regimes, whichsuggest cell membrane disruption as a mechanism of action of the extract.