Time-resolved fluorescence spectroscopy of hematoporphyrin derivative in micelles

Time-resolved fluorescence spectroscopy was performed on hematoporphyrin derivative in buffer at different concentrations of surfactants. New molecular species were detected, with fluorescence decay time constants of ≈0.7 and ≈2.7 ns and emission peaks at 630 and 675 nm, respectively. Deaggregation effects were observed, mainly in the presence of cationic micelles.     

Influence of medium polarity on the photosensitizing properties of hematoporphyrin and hematoporphyrin derivative (Photofrin II)

The influence of the polarity of the medium on the efficiency of Type I and Type II photosensitization as carried out by hematoporphyrin and its derivative Photofrin II was studied by EPR techniques. Our results suggest that porphyrin aggregates do not participate as such in the photosensitized processes. Thus, the solvent-induced breakdown of the aggregated components of Photofrin II appears to determine the photosensitizing efficiency of this porphyrin.     

Porphyrin derivatives having physical and chemical characteristics similar to those of the active components of hematoporphyrin derivative and with very strong photosensitizing effects

The tumour-localizing fraction of hematoporphyrin derivative (Hpd) is thought to possess an essentially diporphyrin ether structure or, alternatively, a diporphyrin ester structure, the properties of which facilitate its retention in malignant cells and its biological activity on irradiation. To elucidate this problem further, we have synthesized the dimethyl, diethyl, dipropyl, di-n-butyl and di-iso-butyl ethers of hematoporphyrin. These ethers show chromatographic properties very similar to those of the active components of Hpd. Furthermore, they are much better photosensitizers in a cellular system than are crude Hpd or Photofrin II, and, like the components of Hpd, they are taken up and retained by cells according to their degree of non-polarity.     

Active oxygen intermediates in the degradation of hematoporphyrin derivative in tumor cells subjected to photodynamic therapy

Hematoporphyrin derivative (HPD), a sensitizer used in photodynamic therapy (PDT) of malignancies, is progressively destroyed during the treatment. Prior studies suggested that upon PDT the photobleaching of HPD in tumor tissues is largely mediated by self-sensitized singlet oxygen. However, little is known about the role of other reactive oxygen species (ROS). The main aim of this work was to clarify the significance of H2O2, superoxide (O2.(-)) and hydroxyl (OH.) radicals in bleaching of HPD in tumor cells subjected to PDT. Experiments were performed on Ehrlich ascites carcinoma (EAC) cells, which were loaded with HPD in PBS and then irradiated with red light at 630 nm in the same buffer. Studies showed that photosensitization of EAC cells by HPD led to the formation of significant amounts of H2O2, O2.(-) and OH., and that these ROS could be involved in the photobleaching of HPD during PDT. In fact, we found that addition of catalase (CAT, a scavenger H2O2), Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and Tiron (scavengers of O2.(-)), Na-benzoate, mannitol and deferoxamine (scavengers of OH.) caused a substantial decrease in the rate of HPD photobleaching in EAC cells. In these cells, the inhibitory effects of Na-benzoate, mannitol and deferoxamine on the photodegradation of HPD correlated well with suppression of the OH. generation, a highly active oxidizer. In EAC cells, the glutathione redox cycle and CAT (scavengers of H2O2) as well as Cu/Zn-SOD was found to suppress the photoinduced degradation of HPD. It was also established that HPD can directly scavenge H2O2 and oxygen free radicals; in a phosphate buffer its second-order rate constants were measured as 5.51+/-0.32 x 10(3)M(-1)s(-1) (for the reaction with O2.(-)), 5.08+/-0.31 x 10(4)M(-1)s(-1) (for H2O2), and 3.44+/-0.08 x 10(10)M(-1)s(-1) (for OH.). Thus, our data suggest that OH. could be one of the main oxidants mediating the photobleaching behavior of HPD in malignancies. Studies showed that photoexcited moieties of HPD can oxidize cell proteins with the formation of protein peroxides (PPO), which currently are regarded as a new form of ROS. Model experiments suggest that PPO could also participate in bleaching of HPD in tumors treated with PDT. It was found that HPD may destroy in tumor cells after cessation of photoirradiation and that this event is largely mediated by the presence of H2O2, a precursor of OH(.).     

Photosensitization of isolated mitochondria by hematoporphyrin derivative (Photofrin): effects on bioenergetics.

Isolated rat liver mitochondria were incubated in the presence of 6 micrograms/ml of Photofrin and irradiated at the wavelength of 365 nm. After 45 s irradiation (30 W/m2), coupling defined as stimulation of respiration by externally added adenosine 5'-diphosphate (ADP) is totally lost. In contrast, membrane potential created by addition of succinate or adenosine 5'-triphosphate (ATP) is only slightly affected. Similarly, the ADP/O ratio is not modified after 20 s irradiation. These data suggest that modification of the mitochondrial membrane potential is not a primary event after irradiation.     

Comparative study on major bioactive components in natural, artificial and in-vitro cultured Calculus Bovis

Major bioactive components in various Calculus Bovis, including natural, artificial and in-vitro cultured Calculus Bovis, were comparatively studied. An approach of high-performance liquid chromatography coupled with ultraviolet and evaporative light scattering detections (HPLC/UV/ELSD) was established to simultaneously determinate six bioactive components thereof, including five bile acids (cholic acid, deoxycholic acid, ursodeoxycholic, chenodeoxycholic acid, hyodeoxycholic acid) and bilirubin. ELSD and UV detector were applied to detect bile acids and bilirubin respectively. The assay was performed on a C(18) column with water-acetonitrile gradient elution and the investigated constituents were authenticated by comparing retention times and mass spectra with those of reference compounds. The proposed method was applied to analyze twenty-one Calculus Bovis extraction samples, and produced data with acceptable linearity, precision, repeatability and accuracy. The result indicated the variations among Calculus Bovis samples under different developmental conditions. Artificial and in-vitro cultured Calculus Bovis, especially in-vitro cultured ones, which contain total bioactive constituents no less than natural products and have the best batch-to-batch uniformity, suffice to be used as substitutes of natural Calculus Bovis.     

NMR study of pradimicin derivative BMY-28864 and its interaction with calcium ions in D2O

The dynamic structure of the antifungal antibiotic pradimicin BMY-28864 in D2O and its interaction with calcium ions were analyzed using one- and two-dimensional 1H nuclear magnetic resonance (NMR). Spectra indicate extensive self-association of molecules in the solution. Two-component spectra were observed simultaneously in a very dilute solution, suggesting equilibrium of two aggregative states. The addition of CaCl2 caused a number of changes in NMR spectra. Therefore we concluded that pradimicin BMY-28864 could form a complex with the Ca2+ ion, causing a movement of the equilibrium. The position of the bound calcium ion is determined indirectly by observing how the NMR shift affects protons that are close to the binding site. The stoichiometry of Ca2+ ion to the Pradimicin molecule for the Ca(2+)-saturated complex is verified to be 1:2. Signal broadening and changes in chemical shift in the 1H NMR spectroscopy of BMY-28864 are assumed to be related to changes in the molecular aggregate conformation.     

Chemical components of the Ephedra major from Iran

Ephedra is a dioecious shrub that belongs to the Ephedraceae family of gymnosperms. Almost all commercial applications of Ephedra extracts are derived from the ephedrine alkaloids found in the evergreen stems. The purpose of this study was to compare chemical components (total alkaloid, ephedrine, total phenol, total flavonoid and tannin) of Ephedra major plants during May to October months. The seeds and stems were collected from Bojnoord altitudes in east of Iran. Total alkaloid was separated by solvent and soxhelet extraction method. The results revealed that solvent extraction method is more efficient than soxhelet extraction method. The measurement of chemical components showed significant difference during May to October months. Data from HPLC analysis revealed that while root is depleted of ephedrine, the ephedrine amount in stem organ ranged from 1.50 ± 0.15 to 2.12 ± 0.01 mg/g dry weight. The results indicate that E. major can be as a suitable source of ephedrine.     

CCQM-P111 study on traceable determination of practical salinity and mass fraction of major seawater components

The first part of the study assessed the equivalence of practical salinity measurement results of a slightly diluted seawater sample from the North Atlantic, which were traceable to the SI. The study shows that the practical salinity reference value S _RV (here S _RV = 34.967) can be determined with a relative standard uncertainty of 3×10⁻⁴. This quantifies the uncertainty range, in which long-term comparability of practical salinity measurement results can be guaranteed currently. In the second part of the study, eight laboratories determined the mass fractions of five seawater components to quantify the equivalence of these measurement results. The results were: Na⁺ (10.726 ± 0.134) g/kg, Mg²⁺ (1.288 ± 0.018) g/kg, Sr²⁺ (0.00755 ± 0.00011) g/kg, Cl⁻ (19.360 ± 0.047) g/kg, SO₄ ²⁻ (2.650 ± 0.025) g/kg. The investigation confirmed the reference composition mass fractions of sodium, magnesium and chloride ions, but it showed a disagreement for strontium and sulphate ions.     

The certification of major components and major elements in five food reference materials

Summary The development of five food reference materials (whole milk powder, pork muscle, rye and wheat flour, and haricot beans) is described. Homogeneity and stability of three categories of nutrients, major components, major elements and vitamins, proved to be adequate. Certification of Kjeldahl nitrogen, total fat, lactose, total dietary fibre (AOAC method), ash, Na, K, Mg, Ca, and Cl contents was successful. In contrast, only indicative values could be given for starch and sugars, nonstarch polysaccharides and P because of insufficient agreement between laboratories. The measurement of these components and elements need further study to obtain the improvement needed. Indicative values for retinol, -carotene, -tocopherol, vitamin B₁, vitamin C, and niacin in some of these materials could be given. Prospects for future certification of vitamins are favourable.     

In vivo derivative NMR spectroscopy for simultaneous improvements of resolution and signal-to-noise-ratio: Case study,Glioma

Journal of Mathematical Chemistry - The theme of this study is derivative nuclear magnetic resonance (dNMR) spectroscopy. This versatile methodology of peering into the molecular structure of...     

Synthesis and study of the spectral properties of diquinone derivatives of hematoporphyrin IX

Diquinone derivatives of hematoporphyrin IX with different structures of the quinone fragments were synthesized by the method of mixed anhydrides. The compounds obtained were investigated by UV, IR, PMR, and fluorescence spectroscopy. Pronounced quenching of the fluorescence of porphyrin in the porphyrin quinones, which depends on the acceptor properties of the quinones and the spatial orientation of the donor and acceptor, was observed.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1324–1330, October, 1991,     

15N and 13C{14N} NMR investigation of the major nitrogen-containing segment in an aquatic fulvic acid: evidence for a hydantoin derivative

A nitrogen-rich segment in a fulvic acid (FA) from Pony Lake, a coastal pond in Antarctica, was investigated by (15)N and (13)C{(14)N} solid-state NMR techniques. As reported previously, the (13)C{(14)N} spectrum of C bonded to N exhibits a peak at 157 ppm that is assigned to an sp(2)-hybridized carbon bonded to at least two nitrogen atoms. This segment contains 48% of all N in the sample. (15)N NMR shows distinct signals, 20 ppm upfield and downfield from the typical peptide resonance; dipolar dephasing confirmed that they are due to protonated N. The well-resolved downfield peak, which accounts for 1/4 of the spectral area, cannot be assigned to aromatic heterocycles, such as purines, because the fraction of aromatic C bonded to N in this sample is very small. Analysis of (15)N chemical-shift trends and (15)N NMR of model compounds, such as arginine and its derivatives, excludes assignment to a guanidinium ion or to substituted guanidino groups. Similarly, ureido groups, -NH-CO-NH-, that are not bonded to a second C = O do not match the observed (15)N peaks in the FA, since both N resonate upfield from the peptide resonance. On the other hand, all chemical shifts are matched within the observed range by the -C(alkyl)-NH-CO-NH-CO-C structure found in two nonaromatic heterocycles, hydantoin and dihydrouracil. The five-membered hydantoin ring, which is found in the purine metabolite allantoin, provides a better match than the six-membered dihydrouracil ring. Regular uracil or thymine fails to produce adequate agreement with observed chemical shifts.     

Possibilities of NMR spectrometry in determining trace components of mixtures

The possibility of using nuclear magnetic resonance (NMR) spectrometry for the determination of trace components in mixtures in the concentration range 1 × 10^?5–5 × 10^?4 M is shown. The conditions of NMR experiments at which the measurement error attains a minimum are standardized and the precision of NMR spectrometry in determining trace components of mixtures within the analytical range is estimated.     

Deuterium incorporation in biomass cell wall components by NMR analysis

A commercially available deuterated kale sample was analyzed for deuterium incorporation by ionic liquid solution (2)H and (1)H nuclear magnetic resonance (NMR). This protocol was found to effectively measure the percent deuterium incorporation at 33%, comparable to the 31% value determined by combustion. The solution NMR technique also suggested by a qualitative analysis that deuterium is preferentially incorporated into the carbohydrate components of the kale sample.     

芘衍生物发光行为的研究

本工作合成了新型的芘衍生物-芘基查尔酮(PD), 研究了其在溶液及胶束中的稳态光物理行为。结果表明: 即使在较低浓度条件下, PD分子间也容易生成基态电荷转移配合物(Inter-CT), 其荧光光谱表现为芘单体荧光、分子内电荷转移态(Intra-CT)荧光以及Inter-CT荧光, 并且具有较好的溶致变色效应、浓度效应以及温度效应, 是一良好的探针分子化合物。     

Identifying the major proteome components of Helicobacter pylori strain 26695

The whole genome sequences of Helicobacter pylori strain 26695 have been reported. Whole cell proteins of H. pylori strain 26695 cells were obtained and analyzed by two-dimensional electrophoresis, using immobilized pH gradient strips. The most abundant proteins were shown in the region of pI 4.0-9.5 with molecular masses from 10 to 100 kDa. Soluble proteins were precipitated by the use of 0-80% saturated solutions of ammonium sulfate. Soluble proteins precipitated by the 0-40% saturations of ammonium sulfate produced similar spot profiles and their abundant protein spots had acidic pI regions. However, a number of soluble proteins precipitated by more than 60% saturation of ammonium sulfate were placed in the alkaline pI regions, compared to those precipitated by 40% saturation. In addition, we have performed an extensive proteome analysis of the strain utilizing peptide MALDI-TOF-MS. Among the 345 protein spots processed, 175 proteins were identified. The identified spots represented 137 genes. One-hundred and fifteen proteins were newly identified in this study, including DNA polymerase III beta-subunit. These results might provide guidance for the enrichment of H. pylori proteins and contribute to construct a master protein map of H. pylori.