HPLC determination of ferrochelatase activity in human liver

A method utilizing HPLC to estimate ferrochelatase activity in human liver cells is presented. A partially purified homogenate of liver cells is incubated with mesoporphyrin IX and cobalt(II)ion. The ferrochelatase in the homogenate incorporates the cobalt(II)ion into the mesoporphyrin. After a fixed time (90 min) the porphyrins are extracted into an ethyl acetate-acetic acid mixture. The porphyrins are then separated using reversed phase HPLC with a mobile phase of methanol-acetonitrile-phosphate buffer pH 3.0 (200:60:30 v/v). The enzyme activity is estimated by measuring the rate of utilization of mesoporphyrin. The optimum pH and substrate concentration for the reaction have been determined.     

Fractionated illumination after topical application of 5-aminolevulinic acid on normal skin of hairless mice: the influence of the dark interval

We have previously shown that light fractionation during topical aminolevulinic acid based photodynamic therapy (ALA-PDT) with a dark interval of 2h leads to a significant increase in efficacy in both pre-clinical and clinical PDT. However this fractionated illumination scheme required an extended overall treatment time. Therefore we investigated the relationship between the dark interval and PDT response with the aim of reducing the overall treatment time without reducing the efficacy. Five groups of mice were treated with ALA-PDT using a single light fraction or the two-fold illumination scheme with a dark interval of 30 min, 1, 1.5 and 2h. Protoporphyrin IX fluorescence kinetics were monitored during illumination. Visual skin response was monitored in the first seven days after PDT and assessed as PDT response. The PDT response decreases with decreasing length of the dark interval. Only the dark interval of 2h showed significantly more damage compared to all the other dark intervals investigated (P<0.05 compared to 1.5h and P<0.01 compared to 1h, 30 min and a single illumination). No relationship could be shown between the utilized PpIX fluorescence during the two-fold illumination and the PDT response. The rate of photobleaching was comparable for the first and the second light fraction and not dependent of the length of dark interval used. We conclude that in the skin of the hairless mouse the dark interval cannot be reduced below 2h without a significant reduction in PDT efficacy.     

Susceptibility to 5-aminolevulinic acid based photodynamic therapy in WHO I meningioma cells corresponds to ferrochelatase activity

5-Aminolevulinic acid (5-ALA) is a natural precursor of protoporphyrin IX (PpIX), which can be used as a photosensitizer in photodynamic therapy (PDT). Accumulation of PpIX in benign meningioma cells has been observed previously, its exploitation for PDT, however, was discouraged by inconsistent results. To evaluate PDT for benign meningiomas, we investigated PpIX synthesis in two human meningioma cell lines (HBL-52 and BEN-MEN-1), their respective extracellular loss of PpIX and corresponding ferrochelatase (FECH) activity as well as their susceptibility to PDT. We demonstrated PpIX production after 5-ALA administration and minor loss to the extracellular space in both cell lines. However, significantly more (up to five times) PpIX was accumulated in BEN-MEN-1 as compared with HBL-52 cells. FECH activity was 2.7-fold higher in HBL-52 compared with BEN-MEN-1 cells and accordingly higher FECH levels could be shown in HBL-52 cells by Western blot analysis. BEN-MEN-1 cells were much more sensitive to PDT and cells could be almost completely killed by irradiation doses of 2 J cm(-2) , whereas HBL-52 showed only an insufficient response at this irradiation dose. We conclude that differences in intracellular PpIX concentrations between HBL-52 and BEN-MEN-1 benign meningioma cells were mainly due to differences in FECH activity and that these differences correspond to their susceptibility to 5-ALA-induced PDT.     

Substituent effect on the photobleaching of pyrylium salts under ultrashort pulsed illumination

The current article presents the photobleaching properties of a group of pyrylium salts under ultrashort pulsed illumination. These pyrylium salts have the same basic chemical structure and differ only by a specific substituent. It is proven experimentally that two different mechanisms are simultaneously present to the photobleaching of all molecules studied (independently of their specific chemical structure). However, the particular parameters of each mechanism are influenced significantly by the substituent change. The experimental investigation of these parameters showed the presence of multiphoton interactions in the photobleaching of pyrylium salts depending essentially on the specific substituent.     

Separation of haem compounds by reversed-phase ion-pair high-performance liquid chromatography and its application in the assay of ferrochelatase activity

The separation of haems and porphyrins was achieved in a reversed-phase ion-pair high-performance liquid chromatography system using tetrabutylammonium hydrogen sulphate as the pairing ion. The concentration of methanol and pH in the mobile phase were determinative parameters for the elution pattern of the compounds. Two isocratic systems--one for the assay of protohaem IX and one for deuterohaem IX--were developed. The chromatographic systems were applied to the assay of ferrochelatase activity in mitochondria using either protoporphyrin or deuteroporphyrin as the substrate. The ferrochelatase activity was also measured in reticulocytes, which contain high levels of endogenous haem.     

Anticancer activity of chamaejasmine: effect on tubulin protein

In this work, the anticancer activity of chamaejasmine was studied by evaluating its in vitro cytotoxicity against several human cancer cell lines (MCF-7, A549, SGC-7901, HCT-8, HO-4980, Hela, HepG2, PC-3, LNCap, Vero and MDCK) using the MTT assay. Results indicated chamaejasmine showed more notable anticancer activity than taxol against PC-3 cells, with IC?? values of 2.28 and 3.98 μM, respectively. Furthermore, Western blot analysis showed that chamaejasmine was able to increase the expression of β-tubulin, but not α-tubulin. In silico simulations indicated that chamaejasmine specifically interacts with the active site which is located at the top of β-tubulin, thanks to the presence of strong hydrophobic effects between the core templates and the hydrophobic surface of the TB active site. The binding energy (E(inter)) was calculated to be -164.77 kcal·mol?1. Results presented here suggest that chamaejasmine possesses anti-cancer properties relating to β-tubulin depolymerization inhibition, and therefore is a potential source of anticancer leads for the pharmaceutical industry.     

The effect of photodynamic therapy (PDT) on oesophageal motility and acid clearance in patients with Barrett's oesophagus

BACKGROUND: Barrett's oesophagus is the major risk factor for oesophageal adenocarcinoma. It is proposed that long-term re-epithelialisation, which has been achieved following ablation using 5-aminolaevulinic acid (5-ALA) photodynamic therapy (PDT) may reduce the risk of malignant change. However, it is not known whether PDT modifies oesophageal motility. AIM: To assess oesophageal pH and motility before and after PDT ablation in treated and untreated areas of the oesophagus. METHODS: Twelve patients (10 male) with Barrett's oesophagus, median segment length 4 cm, were treated with PDT ablation. Twenty-four hours pH assessment and oesophageal manometry were performed before and 4-6 weeks after ablation. PDT was carried out using 635 nm red light, 4-6h after administration of 30 mg/kg 5-ALA. Proximal (untreated) and distal (treated) oesophageal resting pressure, wave amplitude, percentage peristalsis and percentage study time oesophageal pH<4, were assessed. Proton pump inhibitors (PPI) were administered throughout the study. RESULTS: There were no significant differences in oesophageal motility in treated or untreated areas of the oesophagus after PDT compared to pre-treatment values. Patients who continued to experience oesophageal acid exposure required more treatments to achieve complete Barrett's ablation. CONCLUSIONS: Oesophageal motility following ALA-PDT suggests a trend toward enhanced wave propagation however continued oesophageal acid exposure may affect PDT efficacy.     

Indatraline: synthesis and effect on the motor activity of Wistar rats

A new approach for the synthesis of indatraline was developed using as the key step an iodine(III)-mediated ring contraction of a 1,2-dihydronaphthalene derivative. Behavioral tests were conducted to evaluate the effect of indatraline and of its precursor indanamide on the motor activity of Wistar rats. Specific indexes for ambulation, raising and stereotypy were computed one, two and three hours after i.p. drug administration. Indatraline effects on motor activity lasted for at least three hours. On the other hand, no significant differences in motor activity were observed using indanamide. The results suggest that indatraline has a long lasting effect on motor activity and add evidence in favor of the potential use of that compound as a substitute in cocaine addiction.     

Structure effect of benzofuranone on the antioxidant activity in polypropylene

This work deals with the antioxidant activity of benzofuranone compounds in polypropylene (PP). The antioxidant activities of ten benzofuranone compounds in PP were compared using melt flow index (MFI) values of PP stabilized by benzofuranone compounds primarily. The results show: firstly, that the increase of electron donating ability of substituent in 3-substituted benzene ring is beneficial to the improvement of antioxidant activity. Secondly, it has been verified that the steric hindrance of 2′-position substituent can weaken the antioxidant activity of benzofuranone. But when 2′-position substituent forms a hydrogen bond with 3-position reactive hydrogen, the steric hindrance is offset efficiently. Finally, the methyl and tert-butyl groups in the 5 and 7-position of parent benzene ring do not affect the antioxidant activity of benzofuranone compounds in PP obviously.     

Geometric constraints at the surfactant headgroup: effect on lipase activity in cationic reverse micelles

The primary objective of the present article is to understand how the geometric constraints at the surfactant head affect the lipase activity in the reverse micellar interface. To resolve this issue, surfactants were designed and synthesized, and activity was measured in /water/isooctane/n-hexanol reverse micellar systems at z ([alcohol]/[surfactant])=5.6, pH 6.0 (20 mM phosphate), 25 degrees C across a varying range of W0 ([water]/[surfactant]) using p-nitrophenylalkanoates as the substrate. It was observed that lipase activity increases from surfactants to with the increment in surface area per molecule (Amin) because of the substitution by the bulky tert-butyl group at the polar head. However, the activity was found to be similar for despite an enhancement in the hydrophilic moieties at the interface. This unchanged lipase activity is presumably due to the comparable surface area of to originating from the rigidity at the surfactant head. Noticeably, the enzyme activity improved from with the simultaneous increment of both the hydroxyl group and the flexibility of the headgroup whereas that for increased exclusively with the flexibility of the headgroup. The common parameter in both groups of surfactants and is the flexibility of the headgroup, which possibly enhance Amin and consequently the lipase activity. Thus, the geometric constraints at the surfactant headgroup play a crucial role in modulating the lipase activity profile probably because of the variation in interfacial area.     

Lung surfactant dysfunction in tuberculosis: effect of mycobacterial tubercular lipids on dipalmitoylphosphatidylcholine surface activity

In pulmonary tuberculosis, Mycobacterium tuberculosis bacteria reside in the alveoli and are in close proximity with the alveolar surfactant. Mycolic acid in its free form and as cord factor, constitute the major lipids of the mycobacterial cell wall. They can detach from the bacteria easily and are known to be moderately surface active. We hypothesize that these surface-active mycobacterial cell wall lipids could interact with the pulmonary surfactant and result in lung surfactant dysfunction. In this study, the major phospholipid of the lung surfactant, dipalmitoylphosphatidylcholine (DPPC) and binary mixtures of DPPC:phosphatidylglycerol (PG) in 9:1 and 7:3 ratios were modelled as lung surfactant monolayers and the inhibitory potential of mycolic acid and cord factor on the surface activity of DPPC and DPPC:PG mixtures was evaluated using Langmuir monolayers. The mycobacterial lipids caused common profile changes in all the isotherms: increase in minimum surface tension, compressibility and percentage area change required for change in surface tension from 30 to 10 mN/m. Higher minimum surface tension values were achieved in the presence of mycolic acid (18.2 ± 0.7 mN/m) and cord factor (13.28 ± 1.2 mN/m) as compared to 0 mN/m, achieved by pure DPPC film. Similarly higher values of compressibility (0.375 ± 0.005 m/mN for mycolic acid:DPPC and 0.197 ± 0.003 m/mN for cord factor:DPPC monolayers) were obtained in presence of mycolic acid and cord factor. Thus, mycolic acid and cord factor were said to be inhibitory towards lung surfactant phospholipids. Higher surface tension and compressibility values in presence of tubercular lipids are suggestive of an unstable and fluid surfactant film, which will fail to achieve low surface tensions and can contribute to alveolar collapse in patients suffering from pulmonary tuberculosis. In conclusion a biophysical inhibition of lung surfactant may play a role in the pathogenesis of tuberculosis and may serve as a target for the development of new drug loaded surfactants for this condition.     

Influence of yttrium doping on the photocatalytic activity of bismuth oxybromide for ciprofloxacin degradation using indoor fluorescent light illumination

Research on Chemical Intermediates - In this work, a series of Y-doped BiOBr photocatalysts with different yttrium contents were successfully synthesized through a facile ethylene glycol-assisted...     

The effect of illumination on the depth profile of thermally annealed MEH‐PPV/dPS blends

Due to its simplicity and cost‐effectiveness, single solution casting of organic optoelectronics has grown in popularity for device fabrication to produce technologies such as organic photovoltaics and thin film transistors. In order to explore the structural evolution that occurs in the film formation of a blend composed of polystyrene and the benchmark conjugated polymer MEH‐PPV, we have performed a series of neutron reflectivity experiments focused on studying the film structure as it changes through the thermal annealing process both in the presence and absence of white light. Results indicate the formation of a nonhomogeneous blend upon casting, which becomes stratified with thermal annealing. More importantly, the extent of stratification varies with illumination, where exposure to white light increases stratification. This data suggests in situ illumination is a potential novel tool to manipulate device‐relevant morphologies of optoelectronic active layers throughout the fabrication process, offering a cheap nondestructive tool to effectively tune desired structural parameters. © 2017 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2017 , 55, 1142–1149     

Low-temperature synthesis of anatase-brookite composite nanocrystals: the junction effect on photocatalytic activity

Anatase-brookite composite nanocrystals have successfully been synthesized at 50 degrees C using a simple liquid-phase process. The photocatalytic activity of the sample for the gas-phase oxidation of CH3CHO is 5.4 times greater than that of a single-phase anatase sample with comparable crystallite size and surface area. Electron energy loss spectra suggest that this high activity results from junction between anatase and brookite crystals.     

Mechanical treatment effect on the cellulolytic specimen activity

Variation of cellulolytic activity of an enzymatic specimen and of its composition with microcrystallic cellulose upon mechanical treatment has been studied.     

Flavin–cyclodextrin conjugates: effect of the structure on the catalytic activity in enantioselective sulfoxidations

A series of flavin–cyclodextrin conjugates has been prepared and tested in the enantioselective oxidations of prochiral aromatic and aliphatic sulfides with hydrogen peroxide. The newly prepared conjugates contain isoalloxazinium or alloxazinium moieties attached to the primary rim of α- and β-cyclodextrins at the C-6 positions. In addition, flavinium units were attached to the secondary rim of the β-cyclodextrin macrocycle. The relationship between the structural features and the catalytic performance of the conjugates, including those recently reported by us, was analyzed. The rate and enantioselectivity of the sulfoxidations catalyzed by flavin–cyclodextrin conjugates are influenced mainly by the size of the cyclodextrin cavity, the type of flavin unit (alloxazine or isoalloxazine), and by the relative orientation of the flavin and cyclodextrin moieties.     

EPR study of light illumination effects on radicals in gamma-irradiated L-alanine

Exposure of gamma-irradiated L-alanine samples to sunlight and to light from a regular, fluorescent lamp resulted in significant changes in their EPR resonance patterns, both to spectral shapes and intensities. The experimental EPR spectra were numerically decomposed into three components reflecting contributions of three different radicals (R1-R3) generated by ionizing radiation in alanine. The light exposure caused a decay of the measured EPR signal intensity. For similar light intensities and exposure times the decay was much more pronounced in samples illuminated by sunlight than in samples illuminated by the fluorescent lamp. In both cases light-induced decay of R1 radicals was observed. Sunlight illumination resulted in a moderate decay of R2 radicals and in a doubling of the R3 radical population. On the other hand, fluorescent light caused a significant increase of R2 radicals and did not change the amount of R3 radicals. A quantitative analysis of the variations of the three radical contributions to the total EPR spectra upon fluorescent light exposure suggests a net R1-->R2 free radical transformation. These effects of light on the alanine dosimetric signal should be taken into account in dosimetry protocols, assuring protection of alanine dosimeters from extended exposure to light.     

The effect of Ge, Si and Sn phthalocyanine photosensitizers on cell proliferation and viability of human oesophageal carcinoma cells

The photodynamic activity of water soluble mixed sulfonated metallophthalocyanines complexes: GePcSmix, SnPcSmix and SiPcSmix on human oesophageal carcinoma (SNO) cells are reported, and compared with the activity of the unmetallated H2PcSmix and of the newly synthesized water soluble adjacently substituted binaphthalo phthalocyanine (complex 3). The alkaline phosphate (ALP) showed damage to the cell membrane in the presence of complex 3 without irradiation. The GePcSmix complex caused a relatively large increase in inflammation and a high intracellular ATP.     

Solvent effect on surface-enhanced Raman activity of copperphthalocyanine on colloidal silver

Surface enhanced Raman spectrometry (SERS) of copperphthalocyanine (CuPc) in silver hydrosol is reported. The solvent effect on the Raman activity of the adsorbate is discussed in terms of co-adsorption and replacement kinetics of the solvent molecules at the silver surface. It is shown that the quality of the SER spectra can be improved by optimizing the solvent for the adsorbate under study.