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PubMed1992

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↗
PubMed2000

Localization of chromophore absorption signals in TEM with an improved prism-mirror-prism filter.

A corrected prism-mirror-prism electron energy filter with curved entrance and exit faces was designed and incorporated into a Zeiss EM902 transmission electron microscope. The installation of the new filter required little modification to the existing microscope geometry and electronics. The filter had an energy resolution of 1.1 eV over the full image plane (acceptance half angle 10 mradian). The improved energy resolution was applied in studies of the low electron energy loss region that includes molecular orbital excitations or chromophore energy absorptions. Chromophore signal behaviour under electron irradiation was characterized using embedded crystals of hematin and of the dye mercury orange. Images of these crystals confirmed the expected decrease in signal intensity on shifting the selected energy loss from the region of molecular orbital excitations (less than approximately 5 eV) to higher energy losses. Electron irradiation-induced fading of the chromophore signal from hematin and mercury orange yielded similar 1/e dose values of 1.1 x 10(5) e(-) nm(-2) and 1.4 x 10(5) e(-) nm(-2) respectively. In a cellular context, chromophore signals were obtained from energy-filtered images of RIF-1 cell sections containing the photosensitizer chlorin e6 and from sections of BS-C-1 cells with cytoskeletal labelling via FITC-conjugated antibodies. The respective signals were extracted using a dose-dependent method or a shift in selected energy. Chromophore signal distributions were in agreement with fluorescence light microscopic images, but provided detail at higher spatial resolution.

Animals↗
PubMed2002

Interim analysis of a prospective, randomized trial of vacuum-assisted closure versus the healthpoint system in the management of pressure ulcers.

Twenty-eight patients with 41 full-thickness decubitus ulcers were randomized to compare the Vacuum-Assisted Closure device (VAC) with the Healthpoint System (HP) of wound gel products in promoting ulcer healing. A total of 22 patients with 35 full-thickness ulcers completed the 6-week trial of treatment, during which time 2 patients (10%) in the VAC group (N =20) and 2 patients (13%) in the HP group (N = 15) healed completely. The mean percent reduction in ulcer volume was 42.1% with HP and 51.8% with VAC (p = 0.46). The mean number of PMNs and lymphocytes per high-power field decreased in the VAC group and increased in the HP group (p = 0.13, p = 0.41 respectively). The mean number of capillaries per high-power field was greater in the VAC group (p = 0.75). There were 15 cases of biopsy-proven osteomyelitis underlying the ulcers; three (37.5%) improved with VAC and none improved with HP (p = 0.25). VAC promotes an increased rate of wound healing and favorable histological changes in soft tissue and bone compared with HP.

Adult↗
PubMed2004

Effect of chlorophyllin on normothermic storage of human periodontal ligament cells.

The purpose of the present study was to evaluate whether chlorophyllin could serve as an effective constituent of a storage medium to enhance the human periodontal ligament (PDL) cell viability. Freshly isolated PDL cells from premolars extracted from healthy people were stored at 37 degrees C for 6 h in various solutions: F-medium and Hank's balanced salt solution (HBSS), supplemented with chlorophyllin. From MTT viability assays, the highest cell viability was found in the PDL cells stored in HBSS supplemented with 500 nM chlorophyllin, and the chlorophyllin-treated cells showed a dose-dependent response to concentration. Additionally, the results from flow cytometry showed that 77 to 80% of the PDL cells were in the G0/G1 phases of the cell cycle, which suggested that most were in a stable stage. These result showed that HBSS, supplemented with chlorophyllin, may be a useful solution for preserving the viability of PDL cells.

Antioxidants↗
PubMed2002

Localization of rose bengal, aluminum phthalocyanine tetrasulfonate, and chlorin e6 in the rabbit eye.

PURPOSE: The localization and site of action of photosensitizers in the eye may be important for photodynamic therapy for fundus disorders but remain poorly understood for most agents. We investigated the intraocular localization of xanthene, phthalocyanine, and chlorin photosensitizers by using fluorescence microscopy and digital fundus fluorescence angiography. METHODS: Rose bengal (40 mg/kg), aluminum phthalocyanine tetrasulfonate (CASPc) (5 mg/kg), or chlorin e6 (2 mg/kg) was intravenously administered to albino rabbits. The eyes were enucleated and examined by means of fluorescence microscopy 5, 20, 60, and 120 minutes and 24 hours after dye injection. In vivo digital fundus fluorescence angiography with use of rose bengal (2-4 mg/kg), CASPc (2 mg/kg), and chlorin e6 (2 mg/kg) was performed. RESULTS: For all agents studied pathologically, there was moderate fluorescence from the choroid and retinal pigment epithelium 5 minutes after dye injection. Mild fluorescence detected from the photoreceptor outer segments at 5 minutes was increased at 20 minutes. Angiographic studies with use of rose bengal, CASPc, and chlorin e6 revealed differences in the pattern and rate of photosensitizer accumulation. CONCLUSIONS: Rose bengal, CASPc, and chlorin e6 accumulate rapidly in the choroid and retinal pigment epithelium and less rapidly in the outer retina. Differences in ocular localization of these photosensitizers were demonstrated. The significance of these findings for potential photodynamic therapy with these agents requires further investigation.

Animals↗
PubMed2001

Colonic cell proliferation, apoptosis and aberrant crypt foci development in rats given 2-amino-3-methylimidaz.

Sodium-copper chlorophyllin (CHL) inhibits the formation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)- and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced colonic aberrant crypt foci (ACF) and tumours in the F344 rat when it is given simultaneously with either carcinogen. However, CHL reportedly increased the incidence of dimethylhydrazine (DMH)-induced colon tumours in the same species when administered post-initiation. In the present study, rats were given IQ (130 mg/kg body weight, by oral gavages on alternating days) for 2 weeks, starting in experiment week 3, and one week after the final IQ dose rats received CHL treatment until the study was terminated at 16 weeks. Compared with animals given carcinogen alone, the mean number of IQ-induced ACF per colon was reduced significantly by 1% (w/v) CHL in the drinking water (P < 0.05), whereas 0.1% and 0.01% CHL had no effect. These CHL concentrations increased in a dose-related manner both the terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) and bromodeoxyuridine (BrdU) labelling indices in the distal colon. However, the lowest concentration tested, 0.001% CHL, increased the mean number of IQ-induced ACF per colon (P < 0.05), and increased the BrdU labelling index without a concomitant change in TUNEL. These studies indicated that 0.001% CHL promoted IQ-ACF due to deregulation of the homeostatic balance between cell birth and apoptosis in the colonic mucosa, whereas higher concentrations of CHL had either no effect or protected against IQ-induced ACF by causing dose-related increases in the overall rate of cell turnover in the colon.

Administration, Oral↗
PubMedDate not supplied

Relative delivery efficiency and convenience of spray and ointment formulations of papain/urea/chlorophyllin enzymatic wound therapies.

OBJECTIVE: To determine the relative delivery efficiency and user preference for spray and ointment formulations containing papain, urea, and chlorophyllin copper complex sodium (papain/urea/chlorophyllin copper complex). METHODS: Participants applied both a spray and 3 ointment formulations of papain/urea/chlorophyllin copper complex to identical wound models. The average amount of product used per application was determined by weighing the bottle or tube before the study and after completion of the study. Participants were also asked to fill out a questionnaire regarding their preferences for the two formulations. RESULTS: The amount of product used per wound application was 30% less with the spray formulation; resulting in 29% more wound applications per container. Over 80% of the study participants found the spray easier and quicker to use than the ointments. CONCLUSIONS: The spray formulation of papain/urea/chlorophyllin copper complex resulted in less product use per wound application than did 3 different ointment formulations. Participants expressed a favorable impression of practical benefits of the spray formulation including a reduced risk of wound contamination.

Aerosols↗
PubMed1980

Immunopharmacological approach to Forssman shock.

Forssman shock (FS) following the intravenous injection of antisheep erythrocyte antibody into guinea pigs resulted in fatal systemic shock with marked decrease in CH50 values of complement, leucocyte, and platelet counts, and a prolongation of blood coagulation time. In addition, there was an increase in lactate dehydrogenase activity, and decreases in both esterase activity and fibrinogen levels were noted. F(ab')2 of antisheep erythrocyte IgG antibody was not capable of eliciting FS. Cobra venom factor showed a fairly potent inhibition of FS. Leukopenia induced by cytosine arabinoside given intraperitoneally for 5 days had no effect on FS. Colchicine, which decreased the leucocyte count, did not inhibit fatal systemic shock. Administration of heparin or trasyrol did not prevent FS. The present findings demonstrate that FS is inhibited by anticomplementary agents but not by drugs which affect leucocyte and platelet counts, the coagulation system or serum proteases.

Anaphylaxis↗
PubMed1996

Inhibition of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline-mediated DNA-adduct formation by chlorophyllin in Drosophila.

The effect of chlorophyllin on 2-amino-3,8-dimethyimidazo[4,5-f]quinoxaline (MeIQx)-mediated DNA-adduct formation in Drosophila was studied. Third-instar larvae of Drosophila were fed MeIQx at 1 mg/6.5 g-feed/bottle, with or without chlorophyllin (100-300 mg). After a 6 h feeding exposure to MeIQx, the larvae were divided into 2 groups. The first group was examined for covalent DNA adducts by 32P-postlabeling assay. The second group was assayed for DNA damage by allowing the larvae to develop to adults and measuring the male/female ratio (males, DNA repair-deficient; females, DNA repair-proficient). The 32 P-postlabeling results indicated a significant decrease in DNA adduct levels in larvae treated with MeIQx and 300 mg chlorophyllin (1.7 +/- 0.7 adducts/10(7) nucleotides) as compared with MeIQx-treated larvae 6.5 +/- 2.1 adducts/10(7) nucleotides). The results on male/female sex ratios also indicated a chlorophyllin-induced decrease in DNA damage by exposure to MeIQx. The suppressive effect of chlorophyllin on the genotoxic actions of a polycyclic mutagen, MeIQx, may be a result of complex formation between chlorophyllin and the mutagen.

Animals↗
PubMed1996

Inhibitory effect of chlorophyllin on diethylnitrosamine and phenobarbital-induced hepatocarcinogenesis in male F344 rats.

Modifying effects of chlorophyllin (CHL) on the diethylnitrosamine (DEN)-phenobarbital (PB) hepatocarcinogenesis model were examined in rats. Five-week-old male F344 rats were divided into 8 groups. Groups 1 through 5 were given i.p. injections of DEN (100 mg/kg body weight) once a week for 3 weeks beginning one week after the start of the experiment, while groups 6 through 8 received vehicle treatment. Groups 1, 2, 3 and 7 received drinking water with 500 ppm PB from one week after the end of carcinogen or vehicle treatment. CHL-containing diet (2000 ppm) was given to group 2 during the initiation phase and to groups 3 and 5 during the promotion and the post-initiation phase, respectively. Group 6 was given the experimental diet alone throughout the experiment (24 weeks). Liver neoplasms were present in DEN-treated groups and PB treatment promoted liver tumorigenesis. The incidences of adenoma in groups 2 and 3 were significantly smaller than in group 1 (P<0.05 and P<0.02), although the reductions in the incidences of liver cell cancer were not significant. The average numbers of liver neoplasms/rat in group 2 were significantly smaller than in group 1 (P<0.05-P<0.005). Glutathione S-transferase placental form-positive foci were also significantly decreased by CHL treatment (P<0.05 and P<0.001). DEN and PB exposure increased liver ornithine decarboxylase activity and this increase was significantly inhibited by feeding of CHL during the initiation phase (P<0.001). These results suggest that CHL is a chemopreventive agent for liver neoplasia.

Animals↗
PubMed2001

Inhibition of DNA adduct formation and mutagenic action of 3-amino-1-methyl-5h-pyrido[4,3-b]indole by chlorophyllin-chitosan in rpsL transgenic mice.

We have studied the inhibitory effect of chlorophyllin-chitosan (Chl-Chi) complex, an insoluble form of chlorophyllin, on the DNA adduct formation and mutagenesis by a heterocyclic food mutagen-carcinogen, 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), in mice carrying the E. coli rpsL gene as a mutagenesis reporter. Upon administration of a diet containing 0.002% or 0.01% Trp-P-2, DNA adducts were formed in various tissues in a dose-dependent manner, with the maximum level observed in the liver. Addition of 3% Chl-Chi to the diet reduced the Trp-P-2 adduct by up to 90%. The rpsL mutant frequencies increased significantly in both the liver and spleen upon administration of a 0.01% Trp-P-2 diet. Addition of Chl-Chi to the diet decreased these induced mutant frequencies to the background level. No harmful effect of Chl-Chi was detected during these experiments. The results show that Chl-Chi may be a candidate chemopreventive agent against the genotoxic action of Trp-P-2, and possibly also other aromatic carcinogens in the diet.

Animals↗
PubMed2004

Chlorophyllin protects HEp-2 cells from nuclear fragmentation induced by poliovirus.

AIMS: Chlorophyllin (CHLN) is a synthetic derivative of chlorophyll that possesses antimutagenic activity against several environmental contaminants. In the present study, CHLN was assayed for its capacity to prevent nuclear fragmentation (NF) in HEp-2 cells infected with poliovirus. METHODS AND RESULTS: CHLN was assayed at concentrations of 0.5 and 2.5 microg ml(-1), and NF was monitored using the comet assay and acridine orange staining. We demonstrated that CHLN reduced the percentage of NF in poliovirus-infected HEp-2 cells, when cells were treated with drug before infection or exposed continuously to drug. However, the highest degree of protection was achieved when the virus was exposed to CHLN before infection followed by protocol where infected cultures were continuously exposed to the drug after infection. CONCLUSIONS: It is suggested that CHLN primarily binds to the virus which inhibits cell penetration, thereby maintaining nuclear integrity. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering that CHLN has several beneficial properties and no significant toxic effects in humans and animals, it would be an ideal candidate drug to test for antiviral activity.

Antiviral Agents↗
PubMed1980

Use of chlorophyllin in the care of geriatric patients.

In a test group of 62 geriatric nursing home patients, the administration of chlorophyllin tablets was found to be helpful in controlling body and fecal odors. It aided also in easing chronic constipation, served at times as a useful and morale-building placebo, and assisted in abating excessive flatus. No toxic or other ill effects were observed. Bacteriologic studies failed to confirm reports in the literature of antibacterial properties for chlorophyllin. Further investigation of this possibility is recommended.

Aged↗
PubMedDate not supplied

Porphyrin rings and phospholipids: stimulators of cloning efficiency in certain species of Tetrahymena.

Species of Tetrahymena, including T. vorax, T. thermophila, T. pyriformis, and T. pigmentosa, were tested for cloning efficiency in proteose peptone and in synthetic nutrient media to which were added hemin, protoporphyrin IX, chlorophyllin, or asolectin, an impure mixture of phospholipids. All species could be cloned with high efficiency in the crude media. In unsupplemented synthetic medium the cloning efficiencies were 0-10%, around 50%, around 50%, and 90-100% for T. thermophila, T. vorax, T. pyriformis, and T. pigmentosa, respectively. The first three were all stimulated to 90-100% by addition of the porphyrin or phospholipid compounds mentioned above. Uroporphyrin III and coproporphyrin I and III had no effect. We suggest that cells unable to form clones suffer from a lack of cellular energy. This situation may be alleviated by our additions, certain porphyrin rings may be built into cytochromes and phospholipids may be used as fuel. Thus, the synthetic media used so far for these ciliates have not been optimal.

Animals↗
PubMed2004

Photodynamic therapy with chlorin e(6) for skin metastases of melanoma.

BACKGROUND: Photodynamic therapy (PDT) has been successfully applied in clinical settings to destroy neoplasms, but the efficacy of such a treatment is dependent on the type of neoplasm and the photosynthesizer used. Here, we perform a clinical assessment of PDT for skin metastases of pigmented melanoma using chlorin e(6). STUDY DESIGN/MATERIALS AND METHODS: PDT with chlorin e(6) photosensitizer was administered to 14 patients with skin metastases from melanoma (10 females, four males, mean age 49.6 years). Chlorin e(6) at a dose of 5 mg/kg of patient's weight was intravenously injected. The treatment course consisted of two courses of PDT exposure 1 h after intravenous chlorin e(6) injection and 24 h post-injection. The light energy density for each skin tumor was 80-120 J/cm(2) per treatment, with a light power density of 250-300 mW/cm(2). RESULTS: All skin melanoma metastases that received PDT showed complete regression with no recurrence during the study period. The complete response of all skin metastases from melanoma occurred in eight cases after one PDT treatment. In the remaining six individuals, tumors required multiple PDT courses prior to complete regression. No cases of photodermatitis were registered. The Karnofsky performance scale score of the patients with skin metastases from melanoma showed no significant difference before and after PDT. No patients had significant changes in blood cell counts that would indicate chlorin e(6) systemic toxic effect. Blood chemistry and urinalysis did not show any evidence of chlorin e(6) renal and hepatic injury. CONCLUSIONS: PDT with chlorin e(6) for skin metastases from melanoma is effective and well tolerated. Further clinical investigation of PDT with chlorin e(6) is warranted.

Chlorophyllides↗