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Chlorophyll Synthesis in Chlorella II. Effect of Glucose and Light Intensity on the Lag Phase.

Dense cultures of Chlorella grown under our conditions contain large amounts of chlorophyll on a packed cell basis. Upon dilution the cells are, in effect, put into an environment in which light is not limiting and chlorophyll seemingly in excess (at least temporarily). Thus it is not economical for the cell to produce more chlorophyll until it is the limiting factor. It is our contention that the cell may sense this situation by the piling up of products such as glucose or starch. The observed light intensity effect which causes a stimulation of chlorophyll production at lowered intensities is analogous to that previously reported by others and is felt to be related to the rate of photosynthate production.Chlorophyll synthesis commences after a period of about 12 hours. Once it has become established, added substrate or light have a smaller effect. At 36 hours addition of glucose stimulates growth markedly but chlorophyll production continues at the control rate. This suggests that the effect of substrate is not upon the action of an enzyme involved in the actual synthesis of the chlorophyll molecule. Glucose has been known to influence production of certain enzymes involved usually in substrate catabolism. It is quite possible that the role of glucose here is quite similar. The data are consistent with such an explanation. The substrate would have to influence the production of an enzyme in the magnesium branch of the biosynthetic pathway of porphyrin compounds and not in the portion common to both iron and magnesium porphyrins. Accordingly the lag period is viewed as both an inhibitory period as well as a preparatory period for subsequent chlorophyll synthesis.

Journal Article↗

Chlorophyllin is both a positive and negative modifier of mutagenicity.

The mechanism responsible for the modification of mutagenicity by chlorophyllin has been investigated using mutagenic compounds with different mechanisms of action, including the monofunctional alkylating agents, N-methyl-N'-nitrosourea (MNU) and ethylmethanesulphonate (EMS); nitrosamines related to tobacco products, i.e. dimethyl-nitrosamine (DMN), N-nitrosonornicotine (NNN) and 4-(N-methyl-N-nitrosoamino)-1-(3-pyridinyl)-2-butanone (NNK); the polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (B[a]P) and two of its metabolites, i.e. (-)-7 beta,8 alpha-dihydroxy-7,8-dihydrobenzo[a]pyrene (7,8-diol) and (+)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-oxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE); and a complex mutagenic mixture, an extract and subfractions of Swedish moist oral snuff (SMOS). Mutagenicity was monitored with the Ames Salmonella/microsome assays (STY) and hprt V79 point mutation assay (V79). The effects of chlorophyllin on the mutagenicity of the nitrosamines in the STY assays were found to be complex. In the presence of either NNN or NNK, low concentrations of chlorophyllin actually potentiated the mutagenicity > 2-fold. However, at higher, but still non-toxic concentrations, chlorophyllin decreased the mutagenicity of both compounds. The same type of dose-response relationship for chlorophyllin was indicated in the V79 assay system with DMN, although the effect was much weaker. The results with STY were further confirmed by replacing chlorophyllin with another porphyrin compound, hemin. In contrast, biliverdin, a porphyrin structure without the central metal ion, was unable to potentiate the mutagenicity of NNK in STY.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[FTIR studies of chlorobenly-5-fluorouracil porphyrin].

In this paper new kind of porphyrin compounds was prepared, which combined well the affinity of prophyrin to malignant tumour with anticancer of 5-fluorouracil porphyrin. The IR spectra of m- and p-chlorophenyl-5-fluorouracil prophyrin were obtained and analyzed. The IR absorption peaks were investigated and concluded in detail. The IR absorption rule of chloropenly-5-fluorouracil prophyrin which occurred when m-substitution and p-substitution appeared respectively on phenyl, and single substitution of N1 and double substitution of N1 appeared respectively on phenyl or N1 and N1, N3 positions on pyrimidine ring was also discussed. The results show that the relative intensity of nu c-o stretching vibration changed obviously by field-effect when the substitution was strong electro-negative group. Meanwhile, the IR spectral characteristics of the compounds mentioned above were revealed, and this can be used to deduce the stereochemistry structure of them.

Antimetabolites, Antineoplastic↗

Inhibitory effects of chlorophyllin, hemin and tetrakis(4-benzoic acid)porphyrin on oxidative DNA damage and mouse skin inflammation induced by 12-O-tetradecanoylphorbol-13-acetate as a possible anti-tumor promoting mechanism.

Reactive oxygen species (ROS) from both endogenous and exogenous sources can cause oxidative DNA damage and dysregulated cell signaling, which are involved in the multistage process of carcinogenesis such as tumor initiation, promotion and progression. A number of structurally different anticarcinogenic agents inhibit inflammation and tumor promotion as they reduce ROS production and oxidative DNA damage. Evidence suggests that porphyrins can interfere with the actions of various carcinogens and mutagens by forming face-to-face complexes and their antimutagenic or antigenotoxic effects may also be attributed to their antioxidant activities. However, little is known regarding the anti-tumor promoting potential and mechanism of the porphyrin compounds. Based on our previous results on the inhibitory effects of chlorophyllin (CHL), hemin and tetrakis(4-benzoic acid)porphyrin (TBAP) against two-stage mouse skin carcinogenesis, we have investigated their anti-tumor promoting mechanisms. In the present work, CHL, hemin and TBAP reduced superoxide anion generation by 12-O-tetradecanoylphorbol-13-acetate (TPA) in differentiated HL-60 cells and the production of hydroxyl radicals by Fenton reaction. Porphyrins exert a dose-related inhibition of his(+) reversion in Salmonella typhimurium TA102 induced by tert-butylhydroperoxide (t-BOOH). DNA strand breaks by ROS derived from H(2)O(2)/Cu(II) and the formation of 8-hydroxydeoxyguanosine (8-OH-dG) in calf thymus DNA treated with H(2)O(2)/UV also were inhibited markedly by porphyrins in a concentration-dependent manner. Furthermore, CHL, hemin and TBAP decreased myeloperoxidase (MPO) activity and H(2)O(2) formation as well as epidermal ornithine decarboxylase (ODC) activity in mouse skin treated with TPA. These results demonstrate that the antioxidative properties of porphyrins are important for inhibiting TPA-induced tumor promotion.

Animals↗

Electronic interactions and energy transfer in oligothiophene-linked bis-porphyrins.

The photophysical and spectroscopic properties of a series of bis-porphyrin compounds meso-meso-linked via oligothiophene bridges are reported. In particular the effects of the different bridges on the porphyrin properties as well as their ability to enhance energy transfer is investigated. The main findings are: a splitting of the degeneracy of the porphyrin Soret band transition with a lower energy transition aligned along the bridge, a dramatic decrease in triplet lifetime and the occurrence of "superexchange" as the main mechanism for mediating singlet-singlet energy transfer in the case where the bridge is a quaterthiophene. Our results show significant perturbations of the intrinsic porphyrin properties induced by the bridge, which are important for the function of porphyrin assemblies.

Journal Article↗

Vitamins as antimutagens: advantages and some possible mechanisms of antimutagenic action.

Dietary natural inhibitors of mutagenesis and carcinogenesis are of particular importance because they may be useful for human cancer prevention and do not have undesirable xenobiotic effects on living organisms. As was shown in numerous experiments, many endogenous substances, usually obtained in food or synthesized by cells, possess some inhibitory activity towards natural or man-made environmental mutagens which often induce increased frequency of cancer. Among such substances are vitamins, thiol compounds, porphyrin derivatives, polyphenols and others, the antigenotoxicity of which is well established in various genetic tests. Probably a number of these compounds are included in the defense systems of organisms protecting them from harmful exogenous influences continuously affecting genetic material and other components of cells. Some vitamins show protective effects; for example, E, A and C vitamins are active against well-known mutagens both in vitro and in vivo. Genetic properties of other vitamins have been insufficiently explored, but positive results were obtained for a number of them suggesting the desirability of further studies in this field. Synergism of some vitamins activity, both with other vitamins and non-vitamin substances, is of particular interest because clarifying some of their mechanisms of action could be important for understanding the functions of our defense systems.

Anticarcinogenic Agents↗

[The non-trauma fluorescence analysis of the porphyrin in the blood of human superficial skin tissue].

The porphyrin compound in human body mainly includes hemoglobin, protoheme, protoporphyrin, coproporphyrin, uroporphyrin and so on. As we know, the metabolism of the porphyrin in human body depends on the physical conditions. It was found that the relative content of protoporphyrin (PPIX) and carotenoid in blood varied with the developing of the malignant tumor. A system, including Ti: sapphire laser, optical fiber delivery system, fluorescence probe, spectrometer CCD and computer, has been developed to examine the porphyrin in human superficial skin tissue without trauma. It was shown that the fluorescence spectrum of the porphyrin obtained from the superficial skin tissue of human earlobe has the same feature as that from the aqueous solution with PP IX.

Biomarkers, Tumor↗

Excited states of porphyrin isomers and porphycene derivatives: a SAC-CI study.

Excited states of free-base porphyrin isomers, porphycene (Pc), corrphycene (Cor), and hemiporphycene (hPc), were studied by the Symmetry-Adapted Cluster (SAC)/SAC-Configuration Interaction (CI) method. The absorption peaks of the porphyrin isomers were assigned on the basis of the SAC-CI spectra. The X, Y, X', and Y' bands of the porphyrin isomers, which have weak intensities, are identified. The differences in the Q-band absorptions among the isomers were clearly explained by the four-orbital model. In Cor and hPc, the wave function of the B-band corresponds to the mixture of the four-orbital excitations and the optically forbidden excitation of free-base porphin (P), due to the molecular symmetry lowering in the isomers. The B-band character is described by the five-orbital model in Pc and the six-orbital model in Cor and hPc. Two tetrazaporphycenes and two ring-extended (dibenzo) porphycenes were designed, and the Q-band transition moment was successfully controlled. These examples show that the control of the four-orbital energy levels is the guiding principle for pigment design in porphyrin compounds.

Electrochemistry↗

Pheophorbide a, a potent endothelin receptor antagonist for both ETA and ETB subtypes.

Many crude drugs were screened for their capacity to inhibit the binding of endothelin-1 (ET-1) to ET receptors; several crude drugs showed significant binding inhibitory activity. Pheophorbide a (1), a potent non-peptide ET receptor antagonist, was isolated from Altemisiae capillaris Flos ("Inchinko" in Japanese), which has been utilized as a remedy for hepatitis in Oriental medicine. In receptor binding experiments, compound 1 inhibited ET-1 binding specifically to both the ETA receptor (ETAR) and ETB receptor (ETBR), with IC50 values of 8.0 x 10(-8) and 2.1 x 10(-7) M, respectively. Thus, compound 1 is an ET-1 binding inhibitor; however, it exhibited no affinity for the other receptors of angiotensin II and atrial natriuretic peptide. We also evaluated the inhibitory activity of porphyrin compounds, and found that some exhibited moderate activity.

Animals↗

Use of a soft-agar colony-forming assay to determine the photosensitizing effects of several components of hematoporphyrin derivative.

The photosensitizing/cytotoxic effects of four porphyrin compounds derived from hematoporphyrin were compared using two human tumor cell lines with a soft-agar colony-forming assay. Reproducible dose- and time-dependent increases in reproductive cell death were observed for each porphyrin tested. Two fractions derived from hematoporphyrin derivative (HPD) were found to be better photosensitizers than HPD itself, indicating the potential value of this in vitro assay for detecting the most active component from a heterogeneous mixture.

Agar↗

Antioxidant and pro-oxidant effects of a manganese porphyrin complex against CYP2E1-dependent toxicity.

Superoxide dismutases (SOD) mimetics have been shown to be protective against cell injury caused by reactive oxygen species. The objective of this study was to investigate the effects of the manganese (III) tetrakis(N-methyl-2-pyridyl)porphyrin (MnTMPyP) on CYP2E1-dependent toxicity. The synergistic toxicity of iron and arachidonic acid has been associated with oxidative stress and lipid peroxidation in HepG2 cells that overexpress CYP2E1. Iron plus arachidonic acid caused loss of viability, increased lipid peroxidation and reactive oxygen species generation, and mitochondrial membrane injury in these cells. MnTMPyP partially protected against the decrease in cell viability, the enhanced lipid peroxidation and oxygen radical production, and the loss of mitochondrial membrane potential. The effect of MnTMPyP on arachidonic acid (absence of iron) toxicity was also evaluated. Arachidonic acid also caused toxicity, lipid peroxidation and reduction of the mitochondrial membrane potential. However, in this model, all of these alterations were actually enhanced by MnTMPyP. MnTMPyP also enhanced toxicity in CYP2E1-expressing HepG2 cells depleted of reduced glutathione (GSH). MnCl(2) had little or no effect on the toxicity by arachidonic acid, and MnTMPyP itself did not peroxidize arachidonic acid. MnTMPyP, an SOD mimetic that also scavenges hydrogen peroxide and peroxynitrite, thus showed an antioxidant and protective effect against iron plus arachidonic acid toxicity, but a pro-oxidant and cytotoxic effect against arachidonic acid toxicity in CYP2E1-expressing cells. These different actions may relate to the ability of MnTMPyP to either scavenge or produce free radicals in cells depending upon the prevailing MnTMPyP oxidation-reduction pathways. MnTMPyP and related manganese porphyrin compounds may have potential clinical utility against diseases associated with the overproduction of reactive oxygen species such as ethanol-induced liver injury but it is clear that further investigation of all the pathways of manganese porphyrin oxidation-reduction are necessary.

Animals↗

Cell-permeable scavengers of superoxide prevent long-term potentiation in hippocampal area CA1.

Long-term potentiation (LTP) in hippocampal area CA1 is generally dependent on N-methyl--aspartate (NMDA) receptor activation. Reactive oxygen species (ROS), including superoxide, are produced in response to NMDA receptor activation in a number of brain regions, including the hipppocampus. In this study, two cell-permeable manganese porphyrin compounds that mimic superoxide dismutase (SOD) were used to determine whether production of superoxide is required for the induction of LTP in area CA1 of rat hippocampal slices. Incubation of hippocampal slices with either Mn(III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) or Mn(III) tetrakis (1-methyl-4-pyridyl) porphyrin (MnTMPyP) prevented the induction of LTP. Incubation of slices with either light-inactivated MnTBAP or light-inactivated MnTMPyP had no effect on induction of LTP. Neither MnTBAP nor MnTMPyP was able to reverse preestablished LTP. These observations suggest that production of superoxide occurs in response to LTP-inducing stimulation and that superoxide is necessary for the induction of LTP.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Comparative fluorescence spectroscopy shows differences in noncavitated enamel lesions.

The aim of this study was to compare emission spectra from noncavitated enamel caries with different degrees of discoloration under a wide range of excitation wavelengths. Freshly extracted human molars with white spot, light discolored and dark discolored brown spot enamel caries were selected (n = 4 each). Rectangular blocks (3.5 x 3.5 mm) were cut from the carious area and a corresponding sound area of the same tooth. Emission spectra were recorded from carious and the corresponding sound areas using a fluorescence spectrophotometer at excitation wavelengths from 360 nm up to 580 nm in steps of 20 nm. The specimens were submerged in water during measurement. The spectra were corrected and normalized to peak intensity for comparisons between spectra from sound and carious areas of each tooth. Excitation spectra were recorded for selected emission wavelengths that showed maximum intensity. Emission spectra of all types of carious lesions were shifted towards longer wavelengths (red shift) when compared to the spectra of the corresponding sound enamel. The red shift was highest for dark brown spot lesions and lowest for white spot lesions (p <or= 0.001). Distinct fluorescence bands within 600-700 nm typical for porphyrin compounds were strongest for excitation wavelengths from 400 to 420 nm and present in most of the lesions investigated.

Dental Caries↗

252Cf plasma desorption mass spectrometry in the synthesis of porphyrin model systems.

252Cf plasma desorption mass spectrometry has been used in the characterization of more than 100 synthetic porphyrins ranging in mass from 614 u for tetraphenylporphyrin to over 2000 u for some porphyrin model systems. In virtually every case, 252Cf plasma desorption mass spectrometry yielded an intense ionized molecule ion [M.+ and/or (M+H)+], irrespective of the groups appended to the porphyrin. The appended groups include carboxylic acids, amides, imides, chloroacetamides, Fmoc-protected amino acids, aromatic amines, nitriles, alkynes, alkenes, esters, active esters, benzyl ethers, acetals, dithioacetals, ketones, imines, phenols, quinone, hydroquinone, ferrocene, cyanine dyes, trimethylsilyl protecting groups, nitro groups, and combinations of these functionalities. Metalloporphyrins and porphyrin-porphyrin dimers are also analyzed with ease. Resolved isotopic peaks were observed for porphyrins with molecular weights below 1000, and unresolved isotopic peaks yielding average masses were observed for porphyrin compounds with higher molecular weights. The limited resolution in the higher molecular weight range does not lessen the utility of the method because the observation of the molecule ions [M.+ and/or (M+H)+] provides unambiguous evidence concerning the success of the synthesis. The 252Cf plasma desorption mass spectra of porphyrins are not complicated by chemical transformations. This method is ideally suited for rapid analysis of synthetic porphyrins and provides a powerful tool for chemists engaged in the synthesis of complex organic molecules.

Californium↗

Sonodynamically induced antitumor effect of gallium-porphyrin complex by focused ultrasound on experimental kidney tumor.

The antitumor effect of a gallium-porphyrin complex, ATX-70, induced by focused ultrasound, on colon 26 carcinoma implanted in a mouse kidney was investigated. Colon 26 tumors were exposed to focused ultrasound at 500 kHz and 1 MHz in a progressive wave mode. Both frequency components were superimposed onto each other in the focal zone to efficiently produce cavitation in the tumor. ATX-70 was administered intravenously at the dose of 2.5 mg/kg, 24 h before the ultrasonic exposure. Antitumor effects were evaluated by histological observation 7 days after the exposure. The destruction of tumor tissue was observed with the ultrasonic treatment in combination with ATX-70, while neither the treatment with ATX-70 alone nor that with ultrasound alone caused any necrosis. These results first demonstrated that antitumor effects of a porphyrin compound can be induced by focused ultrasound in a progressive wave mode.

Animals↗

Characteristic autofluorescence for cancer diagnosis and its origin.

The autofluorescence spectra of human tissues excited with the 365-nm line of pulsed xenon ion laser have been measured. The autofluorescence spectra of cancer tissues usually show characteristic peaks near 630 nm and 690 nm, which do not appear in the spectra of the corresponding normal tissues. This characteristic fluorescence can be taken as a criterion for cancer diagnosis; a consistency of 89% with the traditional biopsy method has been obtained in preliminary clinical application to the diagnosis of buccal carcinoma. Spectroscopic investigations suggest that the porphyrin compounds localized and retained in cancer tissues might be responsible for this characteristic autofluorescence.

Carcinoma↗

On the possibility of involvement of glutamate:glyoxylate and serine:glyoxylate aminotransferases from rye (Secale cereale L.) seedlings in the metabolism of tetrapyrrole compounds.

The activity of highly purified L-serine:glyoxylate aminotransferase (SGAT, EC 2.6.1.45) from rye seedlings was inhibited competitively by 5-aminolevulinate (ALA, Ki = 5 mM) SGAT was activated by hematin. Protoporphyrin IX and hematin inhibited irreversibly the activity of highly purified glutamate:glyoxylate aminotransferase (GGAT, EC 2.6.1.2) from rye seedlings. SGAT was found to catalyse transamination between ALA and hydroxypyruvate, whereas GGAT that between ALA and 2-oxoglutarate or pyruvate. It is suggested that SGAT is involved in the process of degradation of the excess ALA which has not been incorporated into porphyrin compounds.

Glyoxylates↗

Neocuproine-extended porphyrin coordination complexes. 2. Spectroscopic properties of the metalloporphyrin derivatives and investigations into the HOMO ordering.

The synthesis of a porphyrin compound, 1, containing a 2,9-dimethyl-1,10-phenanthroline moiety that is fused at the beta-pyrrole positions is reported. The absorption spectra of the free-base, copper(II), and zinc(II) derivatives have been studied. On the basis of absorption band intensities, the HOMO of the free base (H21) and its copper and zinc complexes (Cu1 and Zn1) was determined to be of a1u symmetry. Relative to H21, compounds Cul and Znl show enhanced spectral changes upon external metal ion binding. Although the HOMO is the same in all three compounds, the energy gap between the two highest occupied orbitals is greater for Cu1 and Zn1 than it is for the free-base compound. Several metal ions (Ni2+, Cu+, Cu2+, Zn2+, Li+) were examined in their binding to the phenanthrolinic group by measuring the resulting changes in the absorption spectra. It is shown that the observed changes in the absorption spectra are insensitive to the nature of the metal ion coordinated by the phenanthroline moiety. Significant differences in the absorption and emission spectra between Zn1 and [Zn(Zn1)2]2+ clearly demonstrate that the porphyrin pi-system is strongly affected by the binding of metal ions at the fused phenanthrolinic moiety.

Antineoplastic Agents↗