Build-up of esterified aminolevulinic-acid-derivative-induced porphyrin fluorescence in normal mouse skin.
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Aminolevulinic Acid↗
Biomedical subjects
J M Nesland
Publications and source records attributed to J M Nesland.
Apoptosis and necrosis induced with light and 5-aminolaevulinic acid-derived protoporphyrin IX.
The mode of cell death induced by photodynamic treatment (PDT) was studied in two cell lines cultured in monolayer, V79 Chinese hamster fibroblasts and WiDr human colon adenocarcinoma cells. The cells were incubated with 5-aminolaevulinic acid (5-ALA) as a precursor for the endogenously synthesised protoporphyrin IX, which was activated by light. Free DNA ends, owing to internucleosomal DNA cleavage in apoptotic cells, were stained specifically with a fluorescent dye in the terminal deoxynucleotidyl transferase (TdT) assay. The free DNA ends were measured by flow cytometry and the fractions of apoptotic cells determined. Total cell death was measured in a cell survival assay to determine the necrotic fraction after subtraction of the apoptotic fraction. V79 cells did undergo apoptosis while WiDr cells were killed only through necrosis. With time, the apoptotic fraction of V79 cells increased until a maximum was reached about 3-4 h after ALA-PDT treatment. For increasing ALA-PDT doses, a maximal apoptotic fraction 75-85% of the cells was measured at about 85% of total cell death. The flow cytometric assay of apoptosis was confirmed by the typical ladder of oligonucleosomal DNA fragments obtained from agarose gel electrophoresis, by fluorescence micrographs visualising the induced free DNA ends and by electron micrographs showing the typical morphology of apoptotic cells.
Adenocarcinoma↗
Granulocyte colony-stimulating factor improves survival rate and reduces concentrations of bacteria, endotoxin, tumor necrosis factor, and endothelin-1 in fulminant intra-abdominal sepsis in rats.
OBJECTIVE: To study the therapeutic effect of granulocyte colony-stimulating factor (G-CSF) on the mortality rate and host defense pattern in fulminant intra-abdominal sepsis. DESIGN: Prospective, randomized, controlled trial. SETTING: Research laboratory in a university hospital. SUBJECTS: Adult male Wistar rats. INTERVENTIONS: Fulminant polymicrobial intra-abdominal sepsis was induced by a 4-mm cecal perforation. Survival experiments were performed with two different doses of G-CSF (20 and 100 microg/kg/24 hrs), and therapy was started 7 days or 1 day before, or 4 hrs after sepsis induction (n = 24). To examine alterations in host response pattern, G-CSF (20 microg/kg/24 hrs) was given at sepsis induction, and rats were killed 4, 8, 12 and 24 hrs later (n = 8-16 per time period). Histologic examination of lung, liver, spleen, and kidney was performed, and blood concentrations of bacteria, endotoxin, tumor necrosis factor (TNF), endothelin-1, packed cell volume, and lactate were determined. MEASUREMENTS AND MAIN RESULTS: G-CSF (20 microg/kg/24 hrs), given 4 hrs after sepsis induction, reduced the mortality rate from 96% to 42%. Increasing the dose (100 micrograms/kg/24 hrs), or giving G-CSF as prophylaxis (starting 7 days or 1 day before sepsis), gave no further protection. G-CSF attenuated the sepsis-induced enhancement of circulating bacteria, endotoxin, TNF, and endothelin-1, resulting in improved fluid balance and reduced lactate concentration. No histopathologic alterations were observed after G-CSF treatment. CONCLUSIONS: G-CSF improves host defense and survival rate in experimentally induced fulminant intra-abdominal sepsis. Clearance of bacteria and endotoxin is improved, concentrations of TNF and endothelin-1 are suppressed, and microvascular flow is improved. G-CSF does not induce neutrophil-mediated tissue damage.
Animals↗
Uptake, localization, and photodynamic effect of meso-tetra(hydroxyphenyl)porphine and its corresponding chlorin in normal and tumor tissues of mice bearing mammary carcinoma.
By using a chemical extraction assay and confocal laser scanning fluorescence microscopy, the kinetic patterns of uptake, elimination, and localization of meso-tetra(hydroxyphenyl)porphine (m-THPP) and its corresponding chlorin (m-THPC) in tumors and various normal tissues of female C3D2/F1 mice bearing CaD2 mammary carcinoma were studied after an i.p. injection of either 5 mg/kg body weight of m-THPP or 1 mg/kg body weight of m-THPC. Moreover, the histological and ultrastructural alterations of the tumors were evaluated after photodynamic therapy (PDT) with m-THPP or m-THPC. The PDT efficacy with m-THPP and m-THPC was also compared. Both m-THPP and m-THPC had a similar kinetic pattern of distribution in the tumors and most normal tissues examined. The concentrations of the dyes in the tissues peaked at 24-48 h after injection. The peak values of the uptake of m-THPP by the tissues were found to decrease in the following order: spleen > urinary tract > kidney > liver > lung > tumor > heart > skin > muscle > brain. However, higher concentrations of m-THPC were taken up by the tumors than by most of the normal tissues studied except for the liver, urinary tract, and skin. m-THPP was mainly localized in the stroma of the tumors, whereas m-THPC was distributed in both vascular interstitium and neoplastic cells of the tumors. Morphological studies showed that PDT with m-THPP resulted in destructive changes in the microvasculature of the tumors, whereas m-THPC-based PDT destroyed both vascular walls and tumor cells of the tumors. The m-THPP-PDT of the tumors was much less efficient than m-THPC-PDT of the tumors, although the dose of m-THPP used was five times higher than that of m-THPC. m-THPP and m-THPC have different efficiency of sensitizing tumors to photodestruction, although they are similar with respect to hydrophobicity. This is likely due to the differences in their intratumoral localization patterns and in their absorption spectra.
Animals↗
TP53 mutations in prostatic cancer. Analysis of pre- and post-treatment archival formalin-fixed tumour tissue.
The TP53 gene mutation pattern in prostatic cancer was examined in relation to progression and survival, using archival formalin-fixed pre- and post-treatment tumour specimens from 84 prostatic cancer patients. Thirty-four had hormone-sensitive tumours and 50 were hormone-resistant. Six of the 34 (18 per cent) therapy-responding tumours and 19 of the 50 (38 per cent) hormone-resistant tumours showed p53 protein accumulation in the post-treatment specimen. Both pre- and post-treatment specimens from these 25 patients were analysed for mutation of the conserved regions of the TP53 gene (exons 5-8), using constant denaturant gel electrophoresis (CDGE) followed by DNA sequencing. In the post-treatment samples, mutations were detected in three of the six patients with hormone-responsive tumours and in 11 of the 19 patients with hormone-resistant tumours. The three (100 per cent) patients with therapy-responsive tumours with mutations and nine of the 11 (82 per cent) patients with therapy-resistant tumours with mutations died of the disease. Thirteen of the 14 mutations in the post-treatment specimens were transitions, 11 occurring at CpG dinucleotides in which codon 273 was involved in ten. A significantly higher proportion of tumours with mutations were poorly differentiated compared with tumours without mutation (P < 0.04). Our findings indicate that TP53 mutation is a late event in tumour development of the prostate gland and that codon 273 might be a 'hotspot' for mutation in the progression of the disease.
Aged↗
Immunohistochemical detection of nm23/NDP kinase and cathepsin D in medullary carcinomas of the thyroid gland.
Reduced expression of nm23/NDP kinase and increased expression of cathepsin D seem to be correlated with the high metastatic potential in a variety of malignancies. The expression of nm23/NDP kinase and that of cathepsin D have been evaluated by means of an immunohistochemical technique in paraffin-embedded tissues from 44 primary medullary carcinomas of the thyroid gland (MCT) and from the corresponding lymph node metastases in 32 of these cases. In addition, lymph node metastases from 4 cases were studied. We found that 36 of 44 (82%) primary and 26 of 36 (72%) lymph node metastatic MCT were nm23/NDP kinase positive, whereas 14 of the 44 (32%) primary and 17 of the 36 (47%) lymph node metastatic MCT were cathepsin D positive. We found no indication that the nm23/NDP kinase level has any prognostic significance in MCT. The cathepsin D level is close to being prognostically significant in this study, and we cannot exclude the possibility that it could be of prognostic value. However, it seems to be quite weak, and therefore of little use in a clinical situation.
Adolescent↗
SYBR green I DNA staining increases the detection sensitivity of viruses by polymerase chain reaction.
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Benzothiazoles↗
TP53 gene mutations and protein accumulation in primary vaginal carcinomas.
Primary carcinomas from 46 patients were screened for TP53 alterations. Immunohistochemistry demonstrated nuclear TP53 protein accumulation in 22 (48%) cases using the polyclonal CM1 antiserum, whereas 15 (33%) cases showed positive nuclear staining with the mononuclear antibody PAb 1801. Constant denaturant gel electrophoresis (CDGE) was used to screen 27 of the vaginal carcinomas for mutations in the conserved regions of the TP53 gene (exons 5-8). Six of these tumours (22%) contained mutations: four were found in exon 5 and two in exon 8. A total of 50% of the primary vaginal carcinomas carried a TP53 alteration. These results indicate that TP53 abnormalities may be involved in the development of these tumours. However, there was no significant association between TP53 abnormalities and survival.
Adult↗
Growth inhibition of human tumor xenografts in nude mice by treatment with the antitumor agent 4,6-benzylidene-d1-D-glucose (P-1013).
4,6-Benzylidene-d1-D-glucose, P-1013, a deuterated benzaldehyde derivative which acts as a reversible protein synthesis inhibitor in vitro, was evaluated for antitumor effects in two human tumor xenografts implanced s.c. in nude mice. The drug, dissolved in isotonic saline, was given p.o. daily for several weeks. For evaluation of drug efficacy, mean tumor volume growth curves were generated and tumor volume doubling time (TD) as well as per cent change in tumor size for the treated tumors compared to control (T/C) were calculated. P-1013 at 90 mg/kg for 49 days was effective against SK-OV-3 human ovarian carcinoma (T/C 47% at day 40 and 50% increase in TD). P-1013 at 90 mg/kg also suppressed growth of PANC-1 human pancreatic carcinoma in two experiments (T/C 44 and 50%, 20 and 40% increase in TD, respectively). There was no indication of systemic toxicity in mice receiving P-1013. Histological examinations of each tumor showed that P-1013 treatment of pancreatic xenografts reduced tumor volume without inducing greater necrosis than that comparable to respective control tumors. For the ovarian xenograft, the histological examination indicated a higher fraction of tumors with more than 50% necrotic tissue in two of the P-1013-treated groups compared with the control group (Fisher exact test, p = 0.12). It is possible that P-1013, in addition to inhibiting the rate of tumor volume growth, also induces tumor necrotization in the ovarian xenograft.
Animals↗
Alterations at chromosome 17 loci in peripheral nerve sheath tumors.
Little is known about the molecular genetic changes in malignant peripheral nerve sheath tumors (MPNST). Inactivation of the TP53 gene in 17p has been reported in a few tumors. The MPNST is one of the manifestations of neurofibromatosis 1 (NF1), suggesting that the NF1 gene in 17q might be important. We present a study of 15 neurofibromas and MPNST from nine individuals. Seven patients had NF1, and six of these developed MPNST. Genetic alterations at nine polymorphic loci on chromosome 17 were examined. Allelic imbalance was detected only in the malignant tumors from NF1 patients (4/6). Complete loss of heterozygosity of 17q loci was found in three of these tumors, all including loci within the NF1 gene. Two of the malignant tumors also showed deletions on 17p. No mutations were detected within exon 5-8 of the TP53 in any of the MPNST, and none of them were TP53 protein-positive using immunostaining with mono- and polyclonal antibodies against TP53. The numbers of chromosome 17 present in each tumor were evaluated by use of fluorescence in situ hybridization (FISH) on interphase nuclei with a centromere-specific probe. A deviation from the disomic status of chromosome 17 was observed in two of the MPNST from NF1 patients. These results support the hypothesis of inactivation of both NF1 gene alleles during development of MPNST in patients with NF1. In contrast to other reports, we did not find evidence for a homozygous mutated condition of the TP53 gene in the same tumors. Finally, FISH analysis was in accordance with the DNA analysis in the deduction of the numbers of chromosome 17 in these tumors.
Adolescent↗
DNA ploidy, serum prostate specific antigen, histological grade and immunohistochemistry as predictive parameters of lymph node metastases in T1-T3/M0 prostatic adenocarcinoma.
OBJECTIVE: To evaluate whether DNA ploidy and immunohistochemistry performed in primary prostatic carcinoma specimens give predictive information on regional lymph node metastasis in addition to T category, histological grade and serum prostate specific antigen (PSA). PATIENTS AND METHODS: Pre-treatment TURP specimens from 80 patients with prostatic carcinoma T0-T3/M0 disease were retrospectively evaluated by means of DNA ploidy and histological grade, and immunostaining for PSA, prostatic acid phosphatase (PAP), neuron-specific enolase (NSE) and p53 protein. Pelvic lymph node dissection was performed in all patients. Serum PSA was determined in 76 of the 80 patients before pelvic staging lymphadenectomy. Thirty-two (40%) of the 80 patients had pN+ disease. RESULTS: Thirty-six patients (46%) had serum PSA values below the upper reference limit (< or = 10 micrograms/L). By univariate analysis the pN category correlated with the serum PSA level (P < 0.001), histological grade (P < 0.001), tissue PSA (P < 0.001), tissue PAP (P < 0.04), T category (P < 0.005) and DNA ploidy (P < 0.02). Multivariate analysis revealed that the serum PSA level was the most powerful independent prognosticator, followed by the T category, tissue PAP and tissue PSA. Histological grade and DNA ploidy did not reach the level of significance in the multivariate analysis. CONCLUSION: These data suggest that tissue PAP and tissue PSA predict the pN status in patients with T0-T3/M0 prostate carcinoma, in addition to serum PSA and T category. Neuroendocrine differentiation and p53 protein seem to have no predictive ability.
Adenocarcinoma↗
Distribution of 5-aminolevulinic acid-induced porphyrins in noduloulcerative basal cell carcinoma.
Microscopic fluorescence photometry incorporating a light-sensitive thermo-electrically cooled charge-coupled device (CCD) camera was employed to investigate the fluorescence distribution of 5-aminolevulinic acid (ALA)-induced porphyrins in 22 patients with a total number of 52 noduloulcerative basal cell carcinomas (BCC) after topical ALA application with or without dimethylsulfoxide (DMSO)/ethylenediaminetetraacetic acid (EDTA) or after intravenous administration of ALA. Both localization patterns and amounts of ALA-induced porphyrins in the BCC were studied. The ALA-induced porphyrins were localized only in the superficial layers of the noduloulcerative BCC lesions after topical application of 20% ALA alone for 3 h. However, both the penetration of ALA into deep lesions and the production of the ALA-induced porphyrin fluorescence were increased after topical administration of 20% ALA and 20% DMSO/4% EDTA for 3 h. Prior treatment with 99% DMSO for 15 min further enhanced the ALA penetration into the BCC lesions after topical application of the ALA/DMSO/EDTA mixture and produced more ALA-induced porphyrins by a factor of about three compared with those treated with ALA alone. The penetration of ALA into the deep BCC lesions could also be increased by prolonging the time of topical application of 20% ALA/4% EDTA to 29-48 h (without DMSO). Intravenous injection of ALA led to a more homogeneous distribution of the ALA-derived porphyrins in the whole noduloulcerative BCC lesions.
Adult↗
Detection of c-erbB-2 related protein in sera from breast cancer patients. Relationship to ERBB2 gene amplification and c-erbB-2 protein overexpression in tumour.
The level of a c-erbB-2 related protein was determined in sera from 168 breast carcinoma patients, 12 females with benign breast disease, and 66 female controls using an ELISA (enzyme linked immunosorbent assay) kit. Elevated c-erbB-2 related protein level was detected in one of 13 preoperative sera (8%), two of 62 postoperative sera from patients without recurrent disease (3%), and 55 of 93 sera collected at recurrent disease (59%). Elevated serum levels were detected significantly more often in patients with distant metastases than in patients with recurrent disease restricted to loco-regional areas (68% versus 19%). Presence of elevated serum level was associated with ERBB2 gene amplification and c-erbB-2 protein overexpression in tumour. None of the patients who had normal ERBB2 gene copy number in tumour had elevated serum levels. Although the usefulness in postoperative prediction of the presence of micrometastases is somewhat questionable, the results suggest c-erbB-2 related protein to represent a novel tumour marker in serum and other body fluids from breast cancer patients at the time of diagnosis and during treatment monitoring.
Adult↗
Endocrine and metabolic characteristics of polyoma large T transgenic mice that develop ACTH-producing pituitary tumors.
The authors have analyzed several endocrine and metabolic parameters in polyoma large T transgenic mice (PyLT-1) that develop adrenocorticotropic hormone (ACTH)-immunoreactive pituitary tumors and in nontransgenic mice with tumor transplants. All clinically ill PyLT-1 mice (13 to 16 months of age) had pituitary macroadenomas and elevated plasma ACTH levels. Compared to PyLT-1 transgenic mice, the ACTH plasma concentrations in immunocompetent mice with transplant tumors were markedly raised. In these animals, a secondary effect of hypercorticotropism was documented by a moderate hyperglycemia. Furthermore, mice with transplant tumors had a pathological weight increase from the time the tumor was palpable. The present study supports and extends the authors' previous morphological documentation of the similarity between the ACTH-producing tumors in this mouse model and human Cushing's disease.
Adrenocorticotropic Hormone↗
Molecular genetic changes in human male germ cell tumors.
BACKGROUND: An isochromosome for the short arm of chromosome 12, i(12p), is the most common and characteristic cytogenetic aberration in testicular germ cell tumors. Little is known about the molecular genetic abnormalities of these neoplasms. EXPERIMENTAL DESIGN: A total of 32 loci were studied in DNA from 31 primary testicular germ cell tumors and compared with corresponding normal DNA. The loci map to 17 different chromosome arms, including seven that contain known tumor suppressor genes. Southern blot analysis and PCR-based methods were used. Several microsatellite loci were included to investigate instability (seen as new alleles) at repeat loci. The TP53 tumor suppressor gene was analyzed for point mutations by constant denaturant gel electrophoresis and for expression by immunohistochemistry. Histologic sections of the tumor biopsies were evaluated with regard to components and percentage of intact tumor cells. The growth fraction, representing one component of proliferative activity of the tumor, was assessed by the Ki-67 index. RESULTS: Changes were found at all chromosome arms investigated but at very different frequencies, 5-56% of all tumors. The most frequently affected chromosome arms, those showing loss of heterozygosity or allelic imbalance in more than 40% of the tumors, were 2q, 3p, 3q, 11p, 12p, 18q, and 22q. Complete loss of one allele was often seen at 3p and 11p loci, whereas allelic imbalances dominated on the 2p, 3q, 12p, 18q, and 22q loci tested. No mutations were detected within four known mutational hot spots of TP53, but positive immunostaining with two TP53 Ab was seen in 9 of 14 tumors. Most tumors (26 of 31) showed positive immunostaining with Ki-67. Microsatellite instability was not observed. CONCLUSIONS: High frequencies of loss of heterozygosity and allelic imbalance at several loci indicate that inactivation of several tumor suppressor genes may be of importance in developing testicular germ cell tumors. The increase in copynumber of 12p alleles seen in several tumors is likely to reflect one or more 12p isochromosomes. Our findings do not indicate that TP53 plays any major pathogenic role in this tumor type, nor was there any indication that defect repair genes, causing microsatellite instability in other cancers, participate in the progression of testicular cancer.
Alleles↗
Potentiating effects of clofibric acid on the differentiation of HL-60 human promyelocytic leukemia cells induced by retinoids.
Retinoids induce granulocytic differentiation of HL-60 human promyelocytic leukemia cells. In this study we demonstrate that clofibric acid, a well known peroxisome proliferator in rodent, augments the differentiation caused by 0.01 microM and 0.1 microM all-trans retinoic acid, and by 0.3 and 3 microM all-trans retinol. Clofibric acid alone has only marginal effects on the myeloid differentiation, whereas clofibric acid together with the glucocorticoid analogue dexamethasone gives a larger effect. Exposure of cultures to retinoid inhibits the growth of the cells, but despite the increased differentiation addition of clofibric acid to the medium does not further inhibit the growth. HL-60 cells express the nuclear receptor NUCI, a human receptor related to the peroxisome proliferator activated receptor (PPAR).
Cell Differentiation↗
[Molecular pathology. A new diagnostic dimension].
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Humans↗
[Medullary thyroid carcinoma--familial or sporadic disease?].
Medullary thyroid cancer may be autosomal dominantly inherited. Calcitonin is a very sensitive tumour marker in medullary thyroid cancer. It is essential to measure calcitonin in first grade relatives of these patients, in order to expose familial incidents of subclinical disease. In 55 patients with medullary thyroid cancer and no history of familial disease, nine close relatives with elevated calcitonin were identified in four different families. Eight of these nine have been thyroidectomized. Five were found to have medullary thyroid cancer, one had definite C-cell-hyperplasia, and two had equivocal C-cell hyperplasia. The last two, had elevated basal serum calcitonin-levels were increased, but with no further increase after intravenous pentagastrin bolus injection. Thyroidectomy did not modify these results and DNA-analysis may be necessary to draw a conclusion about the hereditary situation of these patients.
Adolescent↗