RESUMO
UNLABELLED: Hypericin, isolated from Hypericum perforatum, is an effective photodynamic substance as demonstrated by various studies. Practical forms of applications of hypericin solutions for systemic use and introduction into body cavities are, however, lacking. We developed an aqueous solution of hypericin non-covalently bound to polyvinylpyrrolidone (PVP). PVP is a poly-N-vinylamide of various degrees of polymerization and forms of intermolecular crosslinks suitable for diagnostic and therapeutic applications. We used PVP (molecular weights of PVP between 10 kD and 40 kD) as a complex forming agent to prepare hypericin for photodynamic therapy and diagnostics. In pure water, hypericin forms aggregates which are non-soluble and non-fluorescent. The hypericin-PVP complex binds more than 1000 mg of hypericin in presence of 100 g PVP or less and is soluble in 1 liter of pure water. Aqueous complex solutions of hypericin-PVP display a characteristic absorption spectrum and fluorescence emission band around 600 nm wavelength. Varying concentrations of hypericin do not cause a blue- or red-shift in the absorption maximum at 595 nm. Excitation at 200 nm to 500 nm leads to emission at 590 nm; a property conducive to diagnostic investigations both in vitro and in vivo. Furthermore, hypericin-PVP exhibits high photostability in the presence of oxygen and broad band light which ensures reproducible photodynamic therapy and diagnosis. CONCLUSION: Hypericin forms liquid molecular chromophore complexes in water when bound to PVP thus allowing investigations in biological media.
Assuntos
Antidepressivos/química , Perileno/análogos & derivados , Antracenos , Carcinoma de Células de Transição/patologia , Fenômenos Químicos , Físico-Química , Sistemas de Liberação de Medicamentos , Citometria de Fluxo , Humanos , Ligação de Hidrogênio , Células K562 , Leucócitos/química , Leucócitos/metabolismo , Perileno/química , Excipientes Farmacêuticos/química , Fotoquímica , Povidona/química , Solubilidade , Espectrometria de Fluorescência , ÁguaRESUMO
Hypericin is a naturally occurring substance found in the common St. John's Wort (Hypericum species) and can also be synthesized from the anthraquinone derivative emodin. As the main component of Hypericum perforatum, it has traditionally been used throughout the history of folk medicine. In the last three decades, hypericin has also become the subject of intensive biochemical research and is proving to be a multifunctional agent in drug and medicinal applications. Recent studies report antidepressive, antineoplastic, antitumor and antiviral (human immunodeficiency and hepatitis C virus) activities of hypericin; intriguing information even if confirmation of data is incomplete and mechanisms of these activities still remain largely unexplained. In other contemporary studies, screening hypericin for inhibitory effects on various pharmaceutically important enzymes such as MAO (monoaminoxidase), PKC (protein kinase C), dopamine-beta-hydroxylase, reverse transcriptase, telomerase and CYP (cytochrome P450), has yielded results supporting therapeutic potential. Research of hypericin and its effect on GABA-activated (gamma amino butyric acid) currents and NMDA (N-methyl-D-aspartat) receptors also indicate the therapeutic potential of this substance whereby new insights in stroke research (apoplexy) are expected. Also in the relatively newly established fields of medical photochemistry and photobiology, intensive research reveals hypericin to be a promising novel therapeutic and diagnostic agent in treatment and detection of cancer (photodynamic activation of free radical production). Hypericin is not new to the research community, but it is achieving a new and promising status as an effective agent in medical diagnostic and therapeutic applications. New, although controversial data, over the recent years dictate further research, re-evaluation and discussion of this substance. Our up-to-date summary of hypericin, its activities and potentials, is aimed to contribute to this process.
Assuntos
Perileno/análogos & derivados , Perileno/farmacologia , Perileno/uso terapêutico , Fitoterapia , Animais , Antracenos , Áustria , Morte Celular/efeitos dos fármacos , Humanos , Estrutura Molecular , Neoplasias/classificação , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Perileno/metabolismo , Fotoquimioterapia/métodos , Fotoquimioterapia/estatística & dados numéricos , Reprodutibilidade dos Testes , Vírus/classificação , Vírus/efeitos dos fármacosRESUMO
Meta-tetrahydroxyphenylchlorin (mTHPC) exhibits significant cytotoxicity against a variety of human cells in culture in combination with light, but also in dark reaction. The ovarian cancer cell line SK-OV3 was incubated with various concentrations of mTHPC and in comparison with Taxol and Cisplatin: then the effect on cell growth was determined. mTHPC exhibited an IC50 of 0.9 muM after 24 hours incubation (IC50 of 1.25 after 2 hours), whereas Cisplatin and Taxol, which, have been used as first line agents for the treatment of ovarian carcinomas, inhibited cell proliferation with an IC50 concentration of 4.6 muM and 78 nM after 24 hours incubation, respectively. Incubation of SK-OV3 cells with mTHPC for 5 days resulted in cytostatic cytotoxicity at a concentration of 0.5 muM. The photodynamic effect of mTHPC depends/among other parameters/on the concentration of the dye present. In combination with light (approximately 15 J/cm2) a linear relationship between the dose of mTHPC and the amount of necrotic cells was observable. Higher concentrations of mTHPC caused necrosis of the ovarian tumor cells. The intracellular concentration of mTHPC showed a linear increase up to 28.6 nM (incubation concentration). In summary, these studies demonstrated that mTHPC exhibits potent antiproliferative activity by inducing necrosis after application of light. MTHPC might be a promising agent with cytostatic and photodynamic properties for the treatment of metastasing ovarian carcinomas. A sensitive PCR method was not able to show the induction of apoptosis in the SK-OV3 ovarian cell line. Using propidium staining, it could be proved that the cell death was caused by necrosis and not through apoptosis after irradiation with light.
Assuntos
Antineoplásicos/farmacologia , Mesoporfirinas/farmacologia , Neoplasias Ovarianas/tratamento farmacológico , Fotoquimioterapia , Apoptose/efeitos dos fármacos , Cisplatino/farmacologia , Feminino , Humanos , Mesoporfirinas/farmacocinética , Necrose , Neoplasias Ovarianas/patologia , Paclitaxel/farmacologia , Células Tumorais CultivadasRESUMO
Recent data strongly suggest tumor cell dissemination of endometrial carcinoma cells in the course of fluid hysteroscopy. In patients who had endometrial cancer which was (except for peritoneal cytology) confined to the uterus, the disease-free survival (DFS) of 135 patients who underwent hysteroscopy prior to staging laparotomy was compared with the DFS of 127 patients without hysteroscopy. After a median follow-up of 23 months, 10 patients experienced tumor recurrence. Although there was a trend towards a higher incidence of positive peritoneal cytology at laparotomy in patients who underwent hysteroscopy, this difference did not achieve statistical significance (P = 0.47). For 5 years, the DFS was 92.4% in patients with hysteroscopy and 84.7% in patients without hysteroscopy before laparotomy (log-rank, P = 0.782). Our data therefore suggest a similar short-term DFS in endometrial cancer patients with and without hysteroscopy prior to laparotomy.
RESUMO
The aim of this study was to treat patients for ectocervical dysplasia [cervical intraepithelial neoplasia (CIN) grades 1 and 2] and associated human papilloma virus (HPV) infections with photodynamic therapy (PDT). In 20 patients, 5-aminolevulinic acid (5-ALA, 12% w/v) was applied topically with a cervical cap 8 h prior to illumination. A thermal light source (150 W halogen lamp) emitting a broadband red light (total energy: 100 J/cm2, fluence rate: 90 mW/cm2) was used for superficial illumination of the portio. In addition, an Nd:YAG pumped dye laser (652 nm) was used to illuminate the cervical canal (total energy: 50 J/cm2, fluence rate: 300 mW/cm2). Preliminary results of follow-ups at 1, 3, 6, and 9 months posttherapy showed a cytological improvement in the grading of the PAP smears in 19 patients and the eradication of cervical HPV in 80%. These results demonstrate that ectocervical dysplasia and associated HPV infections can be treated by PDT.
Assuntos
Ácido Aminolevulínico/administração & dosagem , Papillomaviridae , Infecções por Papillomavirus/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem , Infecções Tumorais por Vírus/tratamento farmacológico , Displasia do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/tratamento farmacológico , Administração Intravaginal , Adolescente , Adulto , Terapia Combinada , Esquema de Medicação , Feminino , Humanos , Infecções por Papillomavirus/complicações , Resultado do Tratamento , Infecções Tumorais por Vírus/complicações , Neoplasias do Colo do Útero/complicações , Displasia do Colo do Útero/complicaçõesRESUMO
Photodynamic therapy involves the application of a photosensitizer activated by visible light to generate cytotoxic reactive oxygen. In addition to clinical investigations, in vitro studies concerning photodynamic potency of sensitizers as well as quantification of illumination procedures are necessary. In our investigation, the objective was to evaluate not only the effects of photosensitizer and light on Gram-positive Staphylococcus aureus, but also to investigate possible synergistic or antagonistic effects of these sensitizers. Therefore, we used hypericin, Photofrin II, porfimer sodium and meso-tetrahydroxyphenylchlorin (mTHPC) alone, as well as in combination. Log-phase cells of S. aureus exhibited a marked sensitivity to white thermal light irradiation in the presence of Photofrin II and mTHPC. However, hypericin caused a rather stimulated growth expressed in increased optical density (OD) and increase of total cell count (TCC) of the culture. Combination sensitization of S. aureus by Photofrin II and mTHPC with hypericin likewise caused a stimulation of bacterial growth. No synergistic effects were obtained by combination of Photofrin II and mTHPC; photoresponse of S. aureus was rather decreased by using combined porphyrins. In comparison, TCC and colony-forming units (CFU) were suppressed in the presence of mTHPC after an illumination procedure as well as in dark reactions. These effects were also obtained in the combination photosensitization by mTHPC and Photofrin II. In the presence the of hypericin, photodynamic effects of mTHPC and Photofrin II were inhibited. It was finally concluded that hypericin in our model is not a proper sensitizer for combination photo-sensitization due to antagonistic effects on photodynamic activity of mTHPC and Photofrin II.
Assuntos
Divisão Celular/efeitos dos fármacos , Luz , Fármacos Fotossensibilizantes/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Técnicas de Cultura de Células , Interações Medicamentosas , Iluminação/métodos , Fatores de TempoRESUMO
We report an unusual case of a nodular variation of pulmonary endometriosis. To our knowledge, there is no previous report on a morphological investigation of this entity. The etiology of this rare condition is still a matter of discussion. The well-circumscribed nodular mass is composed of cells identical to, or closely resembling, those of endometrial stroma containing sparse cysts and glands. Immunohistochemically, the cells showed an extensive co-expression of cytokeratin AE1/AE3 and vimentin and were highly positive for progesterone receptor (PRICA) and estrogen receptor (ERICA). Cells lining the cysts and glands as a monolayer were reactive for Ber-Ep4, cytokeratin Pan and cytokeratin AE1/AE3 and negative to all other markers used including PRICA and ERICA. The differential diagnosis of this entity included fibrous tumor of the pleura and metastatic low-grade-endometrial-stromal-sarcoma. The morphological findings are correlated with immunohistochemical studies and results of cell image analysis. This study details the clinicopathological features of the nodular variation of pulmonary endometriosis.
Assuntos
Neoplasias do Endométrio/patologia , Endometriose/patologia , Pneumopatias/patologia , Sarcoma do Estroma Endometrial/secundário , Adulto , Biomarcadores Tumorais/análise , Cistos/patologia , DNA/análise , Diagnóstico Diferencial , Endometriose/metabolismo , Feminino , Fibroma/patologia , Humanos , Citometria por Imagem , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Pneumopatias/metabolismo , Neoplasias Pleurais/patologia , Ploidias , Células Estromais/metabolismo , Células Estromais/patologia , Tomografia Computadorizada por Raios XRESUMO
This article addresses experimental investigations and the clinical use of PDT in the Rudolfstiftung Hospital, Vienna. We investigated mesotetrahydroxyphenylchlorine (mTHPC) and the photosensitizer hematoporphyrin derivative alone or in combination to prove photodynamic antibacterial effects on Staphylococcus aureus (wild type). mTHPC showed antibacterial toxicity in the dark; hematoporphyrin derivative showed suppressive growth effects only after white-light illumination. Photodynamic activity by the combination of both dyes was obtained in a roughly additive manner. Furthermore, we observed the development of resistance of erythromycin after the illumination procedure with hematoporphyrin derivative. Wild-type S. aureus developed no resistance to the other antibiotics tested. Furthermore, long-term follow-up examinations proved mTHPC-mediated PDT as a possible adjuvant intraoperative therapy in cases of relapses of gynecologic carcinomas. PDT is a tissue-selective and simple intervention. It shows few side effects, and, therefore, it reduces the overall burden of tumor patients. In another clinical investigation, we used 5-aminolevulinic acid-based PDT to treat intraepithelial neoplasia and human papillomavirus of the uterine cervix. 33 of 38 (86,8%) patients with superficial cervical intraepithelial neoplasia grades I and II were treated successfully with PDT. Eradication of human papillomavirus infections was successfully performed in 80% of the cases.
Assuntos
Resistência a Múltiplos Medicamentos , Neoplasias dos Genitais Femininos/tratamento farmacológico , Fotorradiação com Hematoporfirina/métodos , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Animais , Áustria , Quimioterapia Adjuvante , Terapia Combinada , Feminino , Neoplasias dos Genitais Femininos/cirurgia , Humanos , PesquisaRESUMO
The objective of this study was to examine the accuracy of the finding of a histologically well differentiated endometrial carcinoma at dilatation and curettage (D & C) prior to hysterectomy. A retrospective multicentric chart review of 137 endometrial cancer patients was conducted, including all patients in whom a well differentiated endometrial carcinoma had been diagnosed by D & C. Histopathologic grading as determined by D & C was compared with the grading established at the final histologic examination after hysterectomy. Seventy-eight percent of all cases in which a well differentiated tumor was diagnosed with D & C were confirmed as well differentiated endometrial carcinomas, whereas 20.4% had to be upgraded as moderately differentiated tumors after evaluation of the hysterectomy specimen. In one case in which a uterine adenocarcinoma was diagnosed by D & C, a well differentiated adenocarcinoma was found to be combined with a carcinosarcoma in the hysterectomy specimen. In order to avoid false findings of a well differentiated tumor, the histologic grade should be confirmed by intraoperative frozen section examination. This is especially important in cases in which surgical staging was not planned initially.
RESUMO
The efficiency of a new photosensitizer of the second generation, meso-tetra-hydroxyphenyl-chlorin (mTHPC), which has a strong absorption at 652 nm, was investigated by oxygen consumption measurements and membrane integrity testing. The experiments proved a great increase in the efficiency of mTHPC after preincubation at 37 degrees C for 24 hours. From these findings it can be assumed that tumor cells can be treated in an optimal way with PDT after a longer delay following drug administration.
Assuntos
Mesoporfirinas/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Hematoporfirinas , Humanos , Microscopia de Contraste de Fase , Consumo de Oxigênio/efeitos dos fármacosRESUMO
This article addresses the use of meso-tetrahydroxyphenylchlorin-based photodynamic therapy (m-THPC-PDT) to treat recurrent gynaecologic malignancies of the ovary. Photodynamic therapy is an experimental approach in the treatment of neoplasms and results indicate it is a highly tissue selective, relatively simple intervention with few side effects, therefore reducing the overall burden on the patient. Of the three patients involved in the initial study, two were treated solely with photodynamic therapy by laparoscopy, and one underwent additional palliative debulking surgery of metastatic tumours. After a post-operative period of more than two years all three women remained free of relapses.
Assuntos
Antineoplásicos/uso terapêutico , Biomarcadores Tumorais/sangue , Mesoporfirinas/uso terapêutico , Recidiva Local de Neoplasia/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Fotoquimioterapia/métodos , Antígeno Ca-125/sangue , Feminino , Humanos , Recidiva Local de Neoplasia/sangue , Neoplasias Ovarianas/sangueRESUMO
Although rare, sarcoma botryoides of the vagina in infants is a highly aggressive soft tissue tumour. Former opinions suggesting exenterative surgery and radiation to improve the dismal outcome in these young children, are outdated. Due to balanced therapies (topical tumourectomy, chemotherapy and radiation adjusted to tumour stage) cure of these sarcomas may be expected dependent on tumour stage, localisation and response to chemotherapy. The rate of complete remissions has increased in recent years. We report on an eighteen-month old girl with complete remission of tumour stage IIA who had been treated according to the CWS 91 protocol. Diagnosis and treatment were performed in March 1994 at the age of six months. We observed the patient in complete remission for 8 months (till September 1995).
Assuntos
Rabdomiossarcoma Embrionário/congênito , Neoplasias Vaginais/congênito , Biomarcadores Tumorais/análise , Quimioterapia Adjuvante , Terapia Combinada , Feminino , Seguimentos , Humanos , Lactente , Estadiamento de Neoplasias , Rabdomiossarcoma Embrionário/tratamento farmacológico , Rabdomiossarcoma Embrionário/patologia , Rabdomiossarcoma Embrionário/cirurgia , Vagina/patologia , Vagina/cirurgia , Neoplasias Vaginais/tratamento farmacológico , Neoplasias Vaginais/patologia , Neoplasias Vaginais/cirurgiaRESUMO
Studies have shown that meta-tetrahydroxy-phenylchlorin is an efficient tumor targeting agent for laser photodynamic therapy. The effectiveness of this approach for cancer treatment depends on drug concentration, incubation time and extracellular protein. We studied uptake and retention kinetics of mTHPC in a human fibroblast cell line. Our results clearly demonstrate a difference in the amount of extracellular mTHPC at an incubation temperature of 37 degrees C compared to 20 degrees C and 4 degrees C. pH-values were always constant and not responsible for the increase. Furthermore, both absorption and fluorescence of mTHPC increase when incubated at normal human body temperature. Incubation of human fibroblast cells with mTHPC (10 micg/mL) showed that intracellular mTHPC increases in a linear manner reaching saturation after 24 hours and declining until 48 hours with concommitant increase of supernatant mTHPC. Therefore, we believe that tumor cells can be treated optimally with PDT following a delay > 24 hours after drug administration with a minimum of damage to surrounding normal tissues.
Assuntos
Antineoplásicos/farmacocinética , Fibroblastos/metabolismo , Metildopa/análogos & derivados , Fotoquimioterapia/métodos , Radiossensibilizantes/farmacocinética , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Linhagem Celular , Estabilidade de Medicamentos , Fibroblastos/citologia , Humanos , Metildopa/administração & dosagem , Metildopa/química , Metildopa/farmacocinética , Radiossensibilizantes/administração & dosagem , Radiossensibilizantes/química , Sensibilidade e Especificidade , Pele/citologia , Pele/metabolismo , Soluções , Espectrometria de Fluorescência , TemperaturaRESUMO
In a retrospective study of women who had undergone pelviscopic hysterectomies in the last 2 years, we focused on postoperative cyclic spotting. Preoperatively, a regular, cyclic menstruation and a premenopausal sexual hormone constellation were common to all patients. In the first group (n = 29), spotting occurred only in association with two uterine anatomical varieties: uterine hyperflexion and cervical elongation. The second group (n = 18), in which intraoperative internal electrocoagulation of the residual cervical envelope was performed, no spotting occurred despite the presence of uterine hyperflexion and cervical elongation. This procedure prevents the occurrence of postoperative spotting in cases of uterine anatomical varieties.
Assuntos
Histerectomia/métodos , Doenças Uterinas/cirurgia , Útero/anormalidades , Útero/cirurgia , Eletrocoagulação/métodos , Feminino , Humanos , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
A case of pelviscopic hysterectomy with uterine hyperflexion and cervical elongation is presented. In this case we were forced to extirpate the cervical fascia because of uterine hemorrhage by laparotomy after the cervical tissue punch had been performed. The histological findings were supravaginal cervical glands. Complete removal of cervical glandular tissue may be undetermined by hyperflected uteri and cervical elongation.
Assuntos
Colo do Útero/cirurgia , Histerectomia/instrumentação , Laparoscópios , Leiomioma/cirurgia , Metrorragia/cirurgia , Neoplasias Uterinas/cirurgia , Adulto , Colo do Útero/patologia , Fáscia/patologia , Fasciotomia , Feminino , Humanos , Complicações Intraoperatórias/patologia , Complicações Intraoperatórias/cirurgia , Leiomioma/patologia , Metrorragia/patologia , Hemorragia Uterina/patologia , Hemorragia Uterina/cirurgia , Neoplasias Uterinas/patologiaRESUMO
A case of an inflammatory tubo-ovarian tumour is presented. Histology and entomology identified the infectious agent as Phthirus pubis.