Multiphoton excitation fluorescence microscopy of 5-aminolevulinic acid induced fluorescence in experimental gliomas.

Background And Objective: The clinical usefulness of 5-ALA guided detection of tumor tissue has been demonstrated for a number of malignancies. However, current techniques of intraoperative detection of protoporphyrin IX fluorescence in situ do not offer subcellular resolution. Therefore, discrimination of non-specific 5-ALA induced fluorescence remains difficult.

[Fluorescence diagnosis and photodynamic therapy with 5-aminolevulinic acid induced protoporphyrin IX in gynecology: an overview].

Fluorescence diagnosis and photodynamic therapy with 5-aminolevulinic acid induced protoporphyrin IX are promising new options in the diagnosis and therapy of diseases in a wide spectrum of medical disciplines such as urology, dermatology, gastroenterology, surgery, neurosurgery and gynecology. The techniques are based on the application of the heme biomolecule precursor 5-aminolevulinic acid which induces the endogenous accumulation of the photosensitizer protoporphyrin IX in designated tissues. After external excitation with blue light a strong red fluorescence can be initiated particularly in tumorous tissues.

Fluorescence staining of human ovarian cancer tissue following application of 5-aminolevulinic acid: fluorescence microscopy studies.

Background And Objectives: Application of 5-aminolevulinic acid (ALA) for fluorescence-guided second-look laparoscopy has been shown to be a promising new procedure in the early diagnosis of ovarian carcinoma metastases. However, for assessing the reliability of this method, information on the microscopic distribution of protoporphyrin IX (PP IX) in the tissue is needed. Additionally, the selectivity of PP IX uptake is essential for a potential photodynamic therapy (PDT) of ovarian cancer metastases.

Laparoscopic fluorescence detection of ovarian carcinoma metastases using 5-aminolevulinic acid-induced protoporphyrin IX.

Background: The aim of the current clinical study was to evaluate the in vivo fluorescence detection of ovarian carcinoma metastases in a second-look laparoscopic procedure after intraperitoneally applied 5-aminolevulinic acid (ALA).

Methods: Five hours before laparoscopic surgery, ALA was applied intraperitoneally via short infusion in a concentration of 30 mg/kg bodyweight in a sterile, 1% solution. Application of ALA resulted in the endogenous production of the fluorescent photosensitizer, protoporphyrin IX (PP IX).

In-vivo investigations on dye-enhanced photothermal tumor therapy with a naphthalocyanine derivative.

Chromophore-enhanced photothermal therapy involves the application of an exogenous chromophore in combination with irradiation, using an appropriate wavelength, exposure duration and sufficient irradiances. The chromophore palladium(II) octabutoxynaphthalocyanine (PdNc(OBu)8) accumulates at satisfactory concentrations and with good selectivity between both tumor and muscle and tumor and skin in tumor-bearing mice. In an attempt to thermally damage tumor tissue with concurrent sparing of adjacent normal tissue, the potential of PdNc(OBu)8 for photothermal therapy was investigated.

[Lipid metabolism in retinal pigment epithelium. Possible significance of lipoprotein receptors].

Purpose: The retinal pigment epithelium (RPE) regulates the lipid metabolism of the photoreceptors by catalysis of membrane outer segments and via choriocapillary perfusion is also exposed to the regulation of blood lipid levels. Since the uptake the metabolism of cholesterol are mediated by specific low-density lipoprotein (LDL) receptors, expression and regulation of this receptor-type were studied in RPE cultures.

Methods: In vitro experiments were carried out in transformed (SV40) human RPE cells.

Clinical multi-colour fluorescence imaging of malignant tumours--initial experience.

Purpose: The detection of malignant tumours relies on a variety of diagnostic procedures including X-ray images and, for hollow organs, endoscopy. The purpose of this study was to present a new technique for non-invasive tumour detection based on tissue fluorescence imaging.

Material And Methods: A clinically adapted multi-colour fluorescence system was employed in the real-time imaging of malignant tumours of the skin, breast, head and neck region, and urinary bladder.

Photodynamic laser therapy for rheumatoid arthritis. Cell culture studies and animal experiments.

The introduction of arthroscopic techniques has improved the surgical therapy of rheumatoid arthritis. The additional application of the holmium:yttrium aluminum garnet (Ho:YAG) laser likewise holds great promise by providing complete hemorrhagic control. Unfortunately, a minimally invasive solution for use in smaller joints has not yet emerged.

Photodynamic targeting of human retinoblastoma cells using covalent low-density lipoprotein conjugates.

Combination of photosensitizers with carrier molecules has been shown to enhance the efficiency of photodynamic therapy (PDT). Owing to an increased expression of their receptors on some malignant and proliferating cells, low-density lipoproteins (LDLs) are potential endogenous carriers. A photosensitizer, chlorin e6 (Ce6), was covalently bound to LDL via carbodiimide activation.

Chemical instability of 5-aminolevulinic acid used in the fluorescence diagnosis of bladder tumours.

Aqueous solutions of 5-aminolevulinic acid (ALA) prepared for intravesical instillation in the framework of a clinical study on the fluorescence diagnosis of urothelial bladder cancer were found to be unstable. This chemical instability of ALA was studied in aqueous solution of 37 degrees C as a function of concentration, pH and reaction time. Our investigations showed that the reaction of ALA is an irreversible process, which yields at least two reaction products in the pH range studied (pH lower than 8): the 2,5-(beta-carboxyethyl)dihydropyrazine and the 2,5-(beta-carboxyethyl)pyrazine.

Spectral characterization of the benzoporphyrin derivative monoacid ring-A photoproduct formed in fetal calf solutions during irradiation with 694 nm continuous-wave radiation.

Benzoporphyrin derivative monoacid ring A (BPD-MA) is a second-generation photosensitizer for photodynamic therapy (PDT) that has shown good results in phase I clinical trials. Similar to other porphyrin derivatives, BPD-MA readily photobleaches during in-vivo PDT treatment. This study investigated the photodegradation of BPD-MA in fetal calf serum (FCS) solutions in vitro.

[Experimental bases for photodynamic laser therapy in chronic polyarthritis].

The principle of photodynamic laser therapy (PDT) for chronic polyarthritis consists in specifically concentrating a drug (photosensitizer) in the synovium. Subsequent activation of the photosensitizer by means of laser leads to a cytotoxic effect. The practicability of PDT was first shown in cell cultures of human synovial fibroblasts.

Beta-tubulin and P-glycoprotein: major determinants of vincristine accumulation in B-CLL cells.

Vincristine (VCR) accumulation in chronic lymphatic leukemia of B-cell origin (B-CLL) has recently been shown not to be inversely correlated to P-glycoprotein (PGP) levels. Therefore, we studied, in addition to PGP expression and accumulation of VCR, the cellular beta-tubulin content in quiescent and rhIL-2 activated B-CLL cells. VCR mediates cytotoxicity by binding to tubulin.

[Photodynamic therapy of eyelid basalioma after topical administration of delta-aminolevulinic acid].

Delta-aminolevulinic acid (ALA) induced Protoporphyring IX photodynamic therapy (PDT) is a new method in the therapy of cutaneous malignancies. Topical application prevents photosensibilization of normal skin. Promising results in the treatment of superficial basal cell carcinomas have been reported.

Chemomodulation of drugs involved in multidrug resistance in chronic lymphatic leukemia of the B-cell type.

Reduced drug accumulation may be one reason for intrinsic drug resistance in chronic lymphatic leukemia of the B-cell type (B-CLL). Immunophenotyped leukemic human B-cells from 38 cases of B-CLL were characterized for P-glycoprotein (PGP) content. In all, 30 cases of B-CLL were additionally analyzed for further parameters: accumulation of daunorubicin (DNR, n = 20) and rhodamine 123 (Rh123, n = 30) in both the presence and the absence of verapamil (VRP).

MDR hamster cells exhibiting multiple altered gene expression: effects of dexniguldipine-HCl (B859-35), cyclosporin A and buthionine sulfoximine.

An actinomycin D selected, multidrug-resistant (MDR) hamster CHO subline showed strong expression of the P-glycoprotein and sorcin genes together with several other alterations such as a: (i) reduced growth rate, (ii) lowered topoisomerase II, (iii) lowered glutathione-S-transferase-P gene expression, and (iv) the emergence of a 15.5 kDa protein. Besides high resistances to adriamycin, actinomycin D, and vincristine, we observed a lowered sensitivity towards bleomycin, a rather hydrophilic drug usually not involved in P-glycoprotein associated MDR.

Mdr1/P-glycoprotein, topoisomerase, and glutathione-S-transferase pi gene expression in primary and relapsed state adult and childhood leukaemias.

In a variety of adult and childhood leukaemia cell samples collected at different states of the disease, we analysed in a series of sequentially performed slot-blot or Northern-blot hybridisation experiments the expression of genes possibly involved in multiple drug resistance (MDR) (mdr1/P-glycoprotein, DNA topoisomerase II, glutathione-S-transferase pi), and the expression of the DNA topoisomerase I and histone 3.1 genes. Occasionally, P-glycoprotein gene expression was additionally examined by indirect immunocytofluorescence using the monoclonal antibody C219.

Susceptibility of multidrug-resistant human leukemia cell lines to human interleukin 2-activated killer cells.

Considering the possibility to overcome drug resistance by other treatment strategies than chemotherapy we investigated the susceptibility of three independently selected multidrug-resistant sublines of the T-lymphoblastoid leukemic cell line CCRF-CEM to lymphokine-activated killer (LAK) cells. We found that two of the multidrug-resistant sublines were significantly less susceptible targets to LAK cells. A third one, however, was as susceptible as the parental CCRF-CEM cell line.

Changes in clonal growth, immunophenotype, and morphology during a follow-up study of an acute lymphoblastic leukemia.

Cells of a 21-year-old patient with acute lymphatic leukemia were analyzed for morphology and immunophenotype and for genotype consecutively during the course of disease. Initial therapy with the BMFT-ALL protocol (Bundesministerium für Forschung und Technologie) reduced leukemic cells only marginally. The following high-dose Ara-C, mitoxantrone (HAM) chemotherapy led to a cell reduction of 75% and to a drastic change in cell morphology from initially 90% blasts to mainly small lymphoid cells.

Resistance to methotrexate and multidrug resistance in childhood malignancies.

Resistance to drugs, either primary or acquired, is a main problem in cancer chemotherapy. The paper summarizes our results in regard to resistance to methotrexate and multiple drug resistance in human cell lines of pediatric malignancies and in children with resistant cancer. In cell lines as well as in children we could demonstrate amplification of the gene coding for dihydrofolate reductase as a cause for resistance to MTX.

Metabolites of microorganisms. 248. Synthetic analogs of saphenamycin.

A synthesis of racemie saphenic acid is described. From this acid 9 ester derivatives of saphenamycin were prepared. Those with aromatic acid components showed high activity against many Gram-positive and some Gram-negative bacteria.

Expression of a P-glycoprotein gene is inducible in a multidrug-resistant human leukemia cell line.

A human T lymphoblastoid CCRF-CEM cell line exhibiting cross resistance to a variety of drugs was selected with increasing doses of actinomycin D. A subline, designated CCRF ACTD400+, was permanently cultured in the presence of 400 ng/ml Actinomycin D for several months. Using a fragment of the human mdr1 cDNA we found high expression of a 5 kb mRNA species which was not detectable in the sensitive parental CCRF-CEM cell line.

Characterization of actinomycin-D-resistant CHO cell lines exhibiting a multidrug-resistance phenotype and amplified DNA sequences.

Actinomycin D (DACT)-resistant sublines of the Chinese hamster ovary cell line CHO-K1 were selected in vitro. Sublines were derived which expressed 5.2-fold (CHO 15DACT) and 35.

High-dose methotrexate therapy with leucovorin rescue: in vitro investigations on human osteosarcoma cell lines.

High-dose methotrexate (MTX) therapy with subsequent leucovorin (LV) rescue (HDMTX-LV) in the treatment of osteosarcoma is based on the assumption that this tumor has a deficient uptake system for MTX and reduced folates. To simulate features of HDMTX-LV therapy protocols in vitro, sensitive and MTX-resistant human osteosarcoma cell lines and a lymphoblastoid cell line were exposed to MTX and/or LV at various dosages and time schedules and the effects on DNA metabolism and on cell growth were evaluated. The data show that in osteosarcoma cells and in lymphoblasts the cytotoxic effects of 10(-6) M to 10(-7) M MTX can be substantially reversed by LV if the antidote is applied within the first 12 h of MTX exposure.