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PubMed2001

Identification of pelvic lymph nodes with chlorophyllin after injection into the uterine cervix: an experimental and clinical study.

We investigated the value of staining retroperitoneal lymph nodes with chlorophyllin in normal dogs and in women with malignant uterine tumors undergoing lymphadenectomy. In dogs, after 0.3% chlorophyllin (sodium copper chlorophyllin) was injected into the canine uterus, the concentration of dye in the bloodstream was measured with a spectrophotometer and sections of stained retroperitoneal lymph nodes were examined using light and electron microscopy. The highest blood levels were detected at 4 hrs and nearly all of the chlorophyllin was gone from the bloodstream by 18 hrs but was retained in nodal macrophages for at least 4 days. No morphological changes were found in the excised lymph nodes. Twenty-four patients with cervical carcinoma and 20 patients with endometrial carcinoma undergoing radical hysterectomy and lymphadenectomy were divided into a lymphatic coloration group (23 patients) and a non-coloration (control) group (21 patients). In the lymphatic coloration group (0.3% chlorophyllin) was injected into the cervix 5 days before elective lymphadenectomy. There were no complications attributed to injection of the chlorophyllin. The number of dissected lymph nodes in the coloration group were greater than the control group (p<0.01) and the time of operation was shorter (p<0.01). These results suggest that chlorophyllin is safe and facilitates identification of retroperitoneal lymph nodes, allows more complete nodal excision and shortens the time of operation in patients undergoing radical hysterectomy with lymphadenectomy.

Adult↗
PubMed1997

[A study of therapeutic efficacy of Koukangning gargle in the treatment of oral mucositis].

A study of therapeutic efficacy was carried out in 118 patients with oral mucositis (recurrent oral aphthous ulceration, erosive lichen planus, herptic stomatitis and infectious oral mucosal hematoma) at random. As a topical medication, Koukangning gargle was applied to 59 patients of the experimental group, 29 patients of the control group 1 applied Dobell's solution to rinse their mouths, and 30 patients of the control group 2 applied Koutai solution. A satisfactory result was observed in the experimental group, its effectiveness and effective rates rose more than those of control group 1 at both treatments for 3 days and 6 days. Between the two groups, there were markedly significant difference (P < 0.005, P < 0.01, P < 0.005, P < 0.005). Koukangning gargle was nontoxic and no side-effects in the clinical tests. Its efficacy was no significant difference compared with the control group 2. Experimental study demonstrates that it possesses anti-inflamation effect and excellent antibiotic effect, therefore it is a new gargle for rinse mouth combined traditional Chinese medicine with Western medicine.

Anti-Infective Agents, Local↗
PubMed2001

Pegylation of a chlorin(e6) polymer conjugate increases tumor targeting of photosensitizer.

Photodynamic therapy is emerging as a viable modality for the treatment of many cancers. A limiting factor in its use against intracavity tumors such as disseminated ovarian cancer is insufficient selectivity of the photosensitizer for tumor compared with normal tissue. We report on an approach to improve tumor targeting by exploiting differences between cell types and by chemical modification of a photosensitizer conjugate. Attachment of polyethylene glycol (pegylation) to a polyacetylated conjugate between poly-l-lysine and chlorin(e6) increased the relative phototoxicity in vitro toward an ovarian cancer cell line (OVCAR-5) while reducing it toward a macrophage cell line (J774), compared with the nonpegylated conjugate. Surprisingly, the increased phototoxicity of the pegylated conjugate correlated with reduced oxygen consumption. Pegylation also reduced the tendency of the conjugate to aggregate and reduced the consumption of oxygen when the conjugates were illuminated in solution in serum containing medium, suggesting a switch in photochemical mechanism from type II (singlet oxygen) to type I (radicals or electron transfer). Pegylation led to more mitochondrial localization as shown by confocal fluorescence microscopy in OVCAR-5 cells, and, on illumination, produced a switch in cell death mechanism toward apoptosis not seen with J774 cells. Conjugates were injected i.p. into nude mice bearing i.p. OVCAR-5 tumors, and the pegylated conjugate gave higher amounts of photosensitizer in tumor and higher tumor:normal tissue ratios and increased the depth to which the chlorin(e6) penetrated into the peritoneal wall. Taken together, these results suggest that pegylation of a polymer-photosensitizer conjugate improves tumor-targeting and may increase the efficacy of photodynamic therapy for ovarian cancer.

Animals↗
PubMedDate not supplied

[Photoquenching of the bioluminescence of the genetically engineered Escherichia coli TG1 (pXen7) strain in the presence of photodithazine].

The photoquenching of the bioluminescence of the genetically engineered Escherichia coli TG1 (pXen7) strain was studied in the presence of the photosensitizer photodithazine, a glucosamine salt of chlorin e6. The photosensitized quenching of the bioluminescence was found to correlate with the colony-forming ability of the strain. The data obtained are discussed from the standpoint of using biosensor luminescent bacterial systems for the assessment of the efficiency of photosensitizers in antimicrobial photochemotherapy.

Biosensing Techniques↗
PubMedDate not supplied

[The photodynamic inactivation of Candida guilliermondii in the presence of photodithazine].

Photodithazine, a glucosamine salt of chlorin e6, enhanced the inactivation of Candida guilliermondii cells by visible light. The sensitizing effect of photodithazine was found to be related to free or cell surface-bound molecules of this dye. Sodium azide (a singlet oxygen quencher) and propyl gallate (an inhibitor of lipid peroxidation) protected yeast cells from the photodithazine-enhanced photoinactivation.

Antioxidants↗
PubMed1999

[Application of in vitro micronucleus tests using L5178Y mouse lymphoma and WTK1 human lymphoblastoid cell lines to antimutagenicity assay].

In the in vitro micronucleus (MN) tests using L5178Y mouse lymphoma and WTK1 human lymphoblastoid cell lines, 2 well-known antimutagens, Vitamin C (Vc) and the sodium and copper salt of chlorophyll (Chl), were tested to evaluate their ability to inhibit the mutagenicity of mitomycin C(MMC). Three kinds of treatment were designed, namely the simultaneous treatment with Vc (or Chl) and MMC, the use of Vc (or Chl) before the use of MMC, and the use of Vc (or Chl) after the use of MMC. The results showed that Vc and Chl had no MN inductivity in the two cell lines; that simultaneous treatment with Vc and MMC and pre-treatment with Vc effectively reduced MMC-induced MN cell frequencies; and that simultaneous treatment and pre-treatment with Chl at dose level of 0.12 mg/ml, significantly reduced MMC-induced MN cell frequencies of L5178Y cells. The antimutagenic effect of Chl on WTK1 cells was found in simultaneous and pre-treatment at dose levels of 0.12 and 0.06 mg/ml.

Animals↗
PubMed2002

Strategies for chemoprevention of liver cancer.

Liver cancer is one of the most common cancers worldwide. Infection with hepatitis B virus and exposure to aflatoxins in the diet act synergistically to amplify risk. From a public health perspective, hepatitis virus vaccination programs and efforts to both reduce aflatoxin exposures and to attenuate the toxicological consequences of unavoidable exposures should have major impacts on the incidence of this disease. Experimentally, aflatoxin-induced hepatocarcinogenesis can be inhibited by over a score of different chemopreventive agents with multiple mechanisms of action. One agent, oltipraz, is a potent inducer of phase 2 enzymes involved in the detoxication of carcinogens including aflatoxin. A second agent, chlorophyllin, impedes the bioavailability of carcinogens by forming molecular complexes and enhances their elimination in the fecal stream. This review highlights the findings of recent randomized clinical trials with oltipraz and chlorophyllin conducted in individuals exposed to dietary aflatoxins and at high risk for development of liver cancer. Both chemopreventive agents modulated levels of aflatoxin biomarkers in the study participants in manners consonant with protection.

Anticarcinogenic Agents↗
PubMedDate not supplied

[Selectivity of accumulation of chlorine e6 derivatives in blood leukocytes].

The uptake of chlorine e6 derivatives by peripheral blood leukocytes was studied using a fluorescence-activated sorter. The analysis showed that the order of pigment uptake by leukocyte populations is the following: granulocytes > or = monocytes > lymphocytes. It was found that the accumulation of the pigments in the cell significantly varied. The level of chlorine e6 dimethyl ester accumulated by cells was found to be 15 times higher than that of chlorine e6. It was assumed that the differences in pigment uptake by different types of blood cells are due to structural and morphological features of leukocytes. The data obtained may be useful in developing the new methods of photodynamic therapy.

Chlorophyllides↗
PubMed2003

The multidrug resistance modulator SDZ-PSC 833 potentiates the photodynamic activity of chlorin e6 independently of P-glycoprotein in multidrug resistant human breast adenocarcinoma cells.

Photodynamic therapy has clinical indications in treatment of localized cancers and could be interesting for eradication of local recurrence of chemoresistant tumors. In the present study, the intracellular accumulation and distribution of chlorin e6 was investigated in MCF-7 and in P-glycoprotein overexpressing, doxorubicin resistant MCF-7/DXR cell lines. After 3-h incubation with chlorin e6 (1.7 micro mol.l(-1)), no significant difference in accumulation was observed between MCF-7 and MCF-7/DXR cells. Chlorin e6 cellular efflux did not differ in the two cell lines. The lack of influence of P-glycoprotein was confirmed since SDZ-PSC833 (PSC) had no influence in chlorin e6 accumulation and efflux in MCF-7/DXR cells. The subcellular distribution of chlorin e6 appeared different in MCF-7/DXR than in MCF-7 cells. Double staining colocalization fluorescence microscopy studies were performed to identify the subcellular localization sites for chlorin e6 using organelle probes for endoplasmic reticulum, Golgi apparatus, mitochondria and lysosomes. In MCF-7, chlorin e6 was distributed in all cytoplasmic organelles including endoplasmic reticulum and Golgi. In MCF-7/DXR, a diffuse cytoplasmic distribution was observed excepted for the endoplasmic reticulum and Golgi area in which less chlorin e6 was distributed. In MCF-7/DXR, PSC was found to restore the distribution of chlorin e6 in the endoplasmic reticulum and Golgi area while in MCF-7, no effect on the subcellular distribution of chlorin e6 was observed. Although the photodynamic activity of chlorin e6 (1.7 micro mol.l(-1), 650 nm, 8 mW.cm(-2)) was found to be lower in MCF-7/DXR than in MCF-7 cells, PSC was found to potentiate the photodynamic activity of chlorin e6 to similar extent in both cell lines. These results clearly demonstrate that PSC potentiates the photodynamic activity of Chlorin e6 independently of the expression of P-glycoprotein and further suggest that the photodynamic activity of chlorin e6 could be related to its intracellular distribution in the endoplasmic reticulum and the Golgi.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2003

Caspase-8 and apoptosis-inducing factor mediate a cytochrome c-independent pathway of apoptosis in human colon cancer cells induced by the dietary phytochemical chlorophyllin.

Chlorophyllin (CHL), an antimutagenic and anticarcinogenic water-soluble derivative of chlorophyll, was recently found to be highly effective as a chemopreventive agent in a high-risk population exposed unavoidably to aflatoxin B(1) in the diet (P. A. Egner et al., Proc. Natl. Acad. Sci. USA, 98: 14601-14606, 2001). The current study examined the response of HCT116 human colon cancer cells to CHL treatment. Cells exposed to concentrations in the range 0.0625-0.5 mM CHL underwent growth arrest and apoptosis after 24 h, with the formation of a sub-G(1) peak in the attached cell population and nuclear condensation in the floating cell population. There was a concentration-dependent attenuation of mitochondrial membrane potential (deltapsi(m)) without the release of cytochrome c or activation of the caspase-9/caspase-3/poly(ADP-ribose) polymerase pathway. However, apoptosis-inducing factor was released from mitochondria into the cytosol and translocated to the nucleus, leading to concentration-dependent cleavage of nuclear lamins. The upstream mediators of this CHL-induced apoptosis pathway were identified as caspase-8/caspase-6 and truncated Bid, acting in conjunction with other proapoptotic members of the Bcl-2 family, such as Bak. These findings suggest that CHL might trigger apoptosis via interaction with putative "death receptors" in the plasma membrane of cancer cells, leading to initial cleavage of procaspase-8 and activation of subsequent downstream events, resulting in the destruction of nuclear lamins. Importantly, E-cadherin and alkaline phosphatase, which are indicators of cell differentiation, were strongly induced at all concentrations of CHL. Thus, in addition to being an effective blocking agent during the initiation phase, these findings support a role for CHL as a suppressing agent and as a possible novel therapeutic strategy directed toward aberrant cell proliferation in the colon.

Alkaline Phosphatase↗
PubMed2003

Antiproliferative effect of chlorophyllin derived from a traditional Chinese medicine Bombyx mori excreta on human breast cancer MCF-7 cells.

Chlorophyllin (CHL) is the sodium-copper salts of chlorophyll derivatives. In this study, the CHL was derived from a traditional Chinese medicine, the Bombyx mori excreta. The CHL, at concentrations 25-400 microg/ml, reduced the proliferation of HL-60, K-562, S-180, and MCF-7 cells by 8.2-95.7% after 72 h of incubation. The CHL also accumulated G2/M cells and induced apoptosis in the MCF-7 cells. Furthermore, the breast carcinoma cells exhibited lower cyclin D1 and cyclin E levels but higher cyclin B1 level after incubation with the CHL showing that these cyclin changes may be important for the antiproliferative effect of the CHL.

Animals↗
PubMed2003

Selective elimination of corticogeniculate feedback abolishes the electroencephalogram dependence of primary visual cortical receptive fields and reduces their spatial specificity.

The role of corticogeniculate feedback in the organization, function, and state dependence of visual responses and receptive fields (RFs) is not well understood. We investigated the contribution of the corticogeniculate loop to state-dependent changes of characteristics of the primary visual cortex response by using a novel approach of eliminating corticogeniculate projection neurons with targeted neuronal apoptosis. Experiments were performed in anesthetized cats (N2O plus halothane) with parallel recordings of single units from experimental (right) and control (left) hemispheres approximately 2 weeks after induction of apoptosis. Within the experimental hemispheres, neurons of area 17 and of the dorsal lateral geniculate nucleus (dLGN) showed an unusually enhanced and prolonged tonic visual response during episodes of synchronized (syn) EEG activity, whereas response levels during less synchronized states were almost normal. In addition, dLGN cells showed a reduced tendency for burst firing and a less regular spike interval distribution compared with those of controls. These changes are likely attributable to a tonic depolarization of dLGN relay neurons or, more likely, to a decreased responsiveness of thalamic inhibitory processes to cortical feedback. Cortical neurons also displayed an activity-dependent increase in RF size, in contrast to an almost activity-invariant RF size of controls, a phenomenon likely related to the elimination of collateral, intracortical projections of layer 6 neurons. Together, these results demonstrate that selective chronic elimination of corticogeniculate feedback results in the loss of EEG-correlated differences of visual processing in the remaining thalamocortical network, accompanied by a significant increase in excitability during syn EEG, at the expense of noticeably reduced spatial receptive-field specificity in the remaining cortical neurons.

Action Potentials↗