Dietary supplementation of tetradecylthioacetic acid increases feed intake but reduces body weight gain and adipose depot sizes in rats fed on high-fat diets.

AIM: The pan-peroxisome proliferator-activated receptor (PPAR) ligand and fatty acid analogue tetradecylthioacetic acid (TTA) may reduce plasma lipids and enhance hepatic lipid metabolism, as well as reduce adipose tissue sizes in rats fed on high-fat diets. This study further explores the effects of TTA on weight gain, feed intake and adipose tissue functions in rats that are fed a high-fat diet for 7 weeks. METHODS: The effects on feed intake and body weight during 7 weeks' dietary supplement with TTA ( approximately 200 mg/kg bw) were studied in male Wistar rats fed on a lard-based diet containing approximately 40% energy from fat. Adipose tissue mass, body composition and expression of relevant genes in fat depots and liver were measured at the end of the feeding. RESULTS: Despite higher feed intake during the final 2 weeks of the study, rats fed on TTA gained less body weight than lard-fed rats and had markedly decreased subcutaneous, epididymal, perirenal and mesenteric adipose depots. The effects of TTA feeding with reduced body weight gain and energy efficiency (weight gain/feed intake) started between day 10 and 13. Body contents of fat, protein and water were reduced after feeding lard plus TTA, with a stronger decrease in fat relative to protein. Plasma lipids, including Non-Esterified Fatty Acids (NEFA), were significantly reduced, whereas fatty acid beta-oxidation in liver and heart was enhanced in lard plus TTA-fed rats. Hepatic UCP3 was expressed ectopically both at protein and mRNA level (>1900-fold), whereas Ucp1 mRNA was increased approximately 30-fold in epididymal and approximately 90-fold in mesenteric fat after lard plus TTA feeding. CONCLUSION: Our data support the hypothesis that TTA feeding may increase hepatic fatty acid beta-oxidation, and thereby reduce the size of adipose tissues. The functional importance of ectopic hepatic UCP3 is unknown, but might be associated with enhanced energy expenditure and thus the reduced feed efficiency.

Reduced antineoplastic activity in mice of cisplatin administered with high salt concentration in the vehicle.

The effect of high NaCl concentration in the vehicle on the toxic and antineoplastic activities of cisplatin [cis-dichloro-diammineplatinum(II)] (CDDP) was reinvestigated in mice. The toxicity, as measured by the survival of mice given CDDP iv, was reduced by 50-60% when the NaCl concentration in the vehicle was raised from 0.9 to 4%. In ascitic P388 leukemia the antineoplastic activity of CDDP given ip was not reduced significantly. However, in all other systems studied the antitumor activity was reduced when the CDDP was dissolved in high NaCl solution. The tumor models studied included systemic P388 and L1210 leukemias, Lewis lung carcinoma, and 5 human tumor xenografts. The human tumors were studied by the subrenal capsule assay. In the case of a malignant melanoma and a malignant fibrous histiocytoma, the effect also was demonstrated in the subcutaneous nude mouse model. In one of the malignant melanomas a 50% increase in the CDDP dose did not compensate for the reduced antitumor activity caused by the high NaCl concentration in the vehicle. These results, which stand in contrast to current views, question the experimental basis for the use of high-NaCl vehicles in the "high-dose" CDDP regimens.

Up-regulation of the oligosaccharide sialyl LewisX: a new prognostic parameter in metastatic prostate cancer.

Metastatic prostate cancer has an unpredictable long-term prognosis. At present, there are few specific predictors to indicate the outcome for the individual patient. We have studied immunoreactivity for type-2 carbohydrate structures, known to be involved in various cell adhesion processes, in patients with metastatic prostate cancer. One group of patients (n = 26) did not progress within 3 years after orchiectomy, while another group of patients (n = 33) progressed within 1 year following castration and survived less than 2 years. Among the parameters studied, sialyl LewisX carbohydrate up-regulation was the only variable showing significant association with poor prognosis (P < 0.01). Sialyl LewisX discriminated between these two outcome groups with 71% predictability and 96% specificity. Our results indicate that up-regulation of sialyl LewisX is associated with hormonal-resistant, aggressive disease. This prognostic marker may, therefore, have an important role in selecting proper treatment for patients with metastatic prostate cancer.

A characterization of permolybdate and its effect on cellular tyrosine phosphorylation, gap junctional intercellular communication and phosphorylation status of the gap junction protein, connexin43.

Biological and analytical characterizations of permolybdate (a mixture of H2O2 and molybdate) were done. Molybdate (10 mM) and molybdenum(V) chloride (3 mM) did not affect gap junctional intercellular communication (GJIC), phosphorylation status of connexin43 (Cx43) or cellular tyrosine phosphorylation in early passage hamster embryonic cells (mainly fibroblast-like). High concentrations of H2O2 (3-10 mM) affected some of the parameters. Acidified permolybdate was clearly more stable than the unadjusted permolybdate. The maximum biological potency of acidified permolybdate was found at a molar ratio of 2:1 (H2O2:molybdate). The mixtures of molybdenum(V) chloride and H2O2 gave a maximum effect at 4:1 molar ratio (H2O2:molybdenum(V)). This can be explained by decomposition of H2O2 and by the generation of less biologically active compounds. Spectrophotometric analyses of the mixtures corroborated the biological results. The Mo(V) electron spin resonance spectrum disappeared upon addition of H2O2 to Mo(V) solutions, and no spectrum appeared when H2O2 was mixed with Mo(VI). Thus, permolybdate is probably diperoxomolybdate, a Mo(VI) compound. Regardless of the parent metal salt, the H2O2/metal salt mixtures showed concentration-dependent biphasic responses with an initial decrease in GJIC followed by an increase. A dissociation between alteration in Cx43 phosphorylation status and GJIC was obtained under certain conditions. The biological activities of permolybdate were only partially mimicked by phenylarsine oxide, an alternative protein tyrosine phosphatase inhibitor.

A characterization of pervanadate, an inducer of cellular tyrosine phosphorylation and inhibitor of gap junctional intercellular communication.

Gap junctional intercellular communication (GJIC), phosphorylation status of the gap junction protein, connexin43 (Cx43), and cellular tyrosine phosphorylation in Syrian hamster embryo cells have been employed for a biological characterization of pervanadate (a mixture of H2O2 and vanadate). In addition, electron paramagnetic resonance (EPR) spectroscopy was used to follow the appearance and disappearance of vanadyl (V(IV)). It has previously been suggested that pervanadate is vanadyl hydroperoxide (V(4+)OOH). This assumption was tested by using mixtures with different molar ratios of H2O2 and orthovanadate, metavanadate or vanadyl sulfate. The maximal biological activity of the mixtures were found at a molar ratio of 2:1 (H2O2:orthovanadate or metavanadate) or 2.5:1 (H2O2:alkaline vanadyl sulfate). No V(IV) EPR spectrum appeared upon mixing orthovanadate or metavanadate and H2O2. The V(IV) EPR spectrum disappeared when vanadyl sulfate was incubated with H2O2 in a 0.5:1 molar ratio (H2O2:alkaline vanadyl sulfate). Spectrophotometrically, a V(V)-like peak at 265 nm had its optimum at this ratio. These results are consistent with pervanadate being diperoxovanadate. The individual compounds were prominently less active than the per-compound mixtures in affecting the biological parameters. The decreases in GJIC showed a concentration-dependent correlation with the onset of the alterations of the Cx43 band pattern and the amount of phosphotyrosine in cellular proteins, but the correlation was not absolute. All the studied biological parameters were reversible, also under continuous exposure to pervanadate.

The bystander effect in photodynamic inactivation of cells.

Treatment of MDCK II cells with the lipophilic photosensitizer tetra(3-hydroxyphenyl)porphyrin and light was found to induce a rapid apoptotic response in a large fraction of the cells. Furthermore, the distribution of apoptotic cells in microcolonies of eight cells was found to be different from the binomial distribution, indicating that the cells are not inactivated independently, but that a bystander effect is involved in cell killing by photodynamic treatment. The observation of a bystander effect disagrees with the common view that cells are inactivated only by direct damage and indicates that communication between cells in a colony plays a role in photosensitized induction of apoptosis. The degree of bystander effect was higher for cells dying by necrosis than for cell dying by apoptosis.

The treatment of advanced metastatic seminoma: experience in 55 cases.

Thirty-six of 39 previously untreated evaluable patients with advanced metastatic seminoma (stage greater than or equal to IIb) obtained a complete response (CR) and three a partial response (PR) after cisplatin-based combination chemotherapy with or without surgery/radiotherapy (group 1). After a median observation time of 36 months, 33 patients are alive with no evidence of disease (NED). Fifteen additional patients received cisplatin-based chemotherapy due to relapsing seminoma after initial radiotherapy (group 2). Thirteen patients obtained a CR, and two a PR. Patients from group 1 lived significantly longer after the start of chemotherapy than those from group 2. The rate and severity of the treatment-related complications were comparable in both groups. The most frequent nonfatal side effects of cisplatin-based combination chemotherapy were Raynaud-like phenomena, polyneuropathy, and myelosuppression. Four patients developed fatal complications (septicemia, bone marrow aplasia). In three of 12 patients with evaluable postchemotherapy histology, vital malignant seminoma was found. Cisplatin-based combination chemotherapy in advanced seminoma patients is highly effective, but includes a significant risk of severe complications. Less toxic treatment regimens should be explored, at least for patients who have less advanced tumors (stage IIb/limited stage IIc).

Myelopathy following radiotherapy of bronchial carcinoma with large single fractions: a retrospective study.

A series of 387 patients with bronchial carcinoma treated with a concentrated split course regimen was surveyed for radiation myelitis. The total dose was 38 Gy. Three fractions of 6 Gy were given the first week. After a three-week interval the patients had a second course of radiotherapy with fractions of 4 Gy on 5 consecutive days. A spinal shield was used in 230 patients for the last two fractions of radiotherapy. The median survival for all patients was 9.4 months. Seventeen cases of radiation myelopathy were found. The life table method was used to calculate the risk of myelopathy in surviving patients, showing risk levels of 30 +/- 15% in patients surviving three years or more. The average age of the patients with myelopathy was 57.6 years, compared to 62.9% years in the total population. Statistical analysis showed a borderline significant increased risk for myelopathy in females, and a significant increased risk in younger patients (P = 0.03). No difference in the incidence of myelopathy was found in patients treated with or without a spinal shield.

Incidence and control of occult neck node metastases from squamous cell carcinoma of the anterior two-thirds of the tongue.

Cervical lymph node metastases developed in 45% of patients with T1N0 squamous cell carcinomas of the oral tongue in spite of local control of the primary lesions in 79%. The control rate for the neck of those who converted from a negative to a positive neck (N0-N+) was 33%. Neck node metastases developed in 49% of patients with T2N0 tumors. The control rate of the primary tumor was 32%, and the control rate of the neck of those whose neck nodes converted (N0-N+) was 16%. Neck node metastases developed in 42% of patients with T3N0 tumors. The control rate of the primary tumors was 33%, and the control rate of the neck of those who converted (N0-N+) was 7%. The development of neck node metastases in patients after treatment of the primary tongue carcinoma is of grave prognostic significance. The use of elective treatment to the neck at initial treatment can prevent metastases in the neck from developing if the primary tumor is under control.

Salivary gland malignant neoplasms: treatment and prognosis.

A retrospective analysis of 183 patients with malignant salivary gland tumors treated between 1955 and 1978 is presented. The analysis showed that radiation therapy lowered the recurrence rates after surgery and controlled approximately one-third of the inoperable tumors. A dose-response relationship exists and the data suggest that the radiation dose should not be less than that corresponding to a CRE-value of 1950 reu (70 Gy/7 weeks). Histology, location and clinical stage are important prognostic factors.

Radiotherapy for testicular seminoma stage I: treatment results and long-term post-irradiation morbidity in 365 patients.

After infradiaphragmatic radiotherapy the cancer-related 10 year survival was 99% in 365 patients with seminoma Stage I referred to the Norwegian Radium Hospital between 1970 and 1982. Thirteen patients relapsed, 11 of them within the first 3 years after treatment. Nine of the recurrent patients were cured by radiotherapy alone (4) or in combination with chemotherapy (5). There is no need to include the inguinal lymph nodes into the irradiation field or to give scrotal irradiation, not even to patients with tumor infiltration beyond the testicular tissue, or to those with prior scrotal or inguinal surgery. At least 1 year after radiotherapy moderate or more severe dyspepsia was observed in 16 patients. Nine patients developed a peptic ulcer. In general, there was no increased risk for development of a second non-germ cell cancer after radiotherapy. However, 4 patients developed a pulmonary cancer indicating a border-line significance of increased risk for this type of malignancy. (p:0.05). In conclusion, infradiaphragmatic radiotherapy remains the optimal routine treatment in seminoma patients with Stage I.

Bleomycin and radiation therapy in squamous cell carcinoma of the upper aero-digestive tract: a phase III clinical trial.

A group of 222 consecutive patients admitted with squamous cell carcinoma of the upper aero-digestive tract were studied in a prospectively randomized and stratified clinical trial. One-half of the patients received bleomycin injected intramuscularly 1 hour before the radiation treatment daily for 5 days a week; the other half received radiation therapy without the added chemotherapy. The total dose of radiation in both groups was about the same, and was given with curative intent even to the patients with advanced tumors who constituted the majority in both groups. Interstitial radiation as boost therapy or surgery was added in patients with residual tumor if the lesions were considered operable and the patient's condition would allow surgery. The addition of bleomycin did not increase the combined local and regional tumor control rates nor did it improve the survival, but did significantly increase the morbidity and the complication rate.

T2/T3 bladder carcinomas treated with definitive radiotherapy with emphasis on flow cytometric DNA ploidy values.

From 1972 through 1982, 124 patients with muscle invasive T2/T3 bladder carcinoma without evidence of distant metastases were treated with definitive radiotherapy (60 Gy in 6 weeks) to the pelvis at The Norwegian Radium Hospital. Sections from paraffin embedded biopsies taken prior to the start of treatment were prepared for DNA flow cytometry, and 121 histograms were considered interpretable for DNA ploidy values. Significantly improved survival was seen in patients with the following characteristic, univariate analysis: T2 tumor, macroscopically complete removal of the tumor prior to radiotherapy, clinically complete response to radiotherapy, normal serum creatinine, absence of hydronephrosis, tetraploid tumor, WHO performance status 0 or 1, age below 65 years. In a multivariate analysis the following pretreatment variables turned out with prognostic power in this order: normal serum creatinine, T2 tumor, tetraploidy, and absence of small vessel invasion. When including response to therapy, the following factors were significantly predictive of a beneficial survival: clinically complete response after radiotherapy, normal creatinine, T2 tumor and tetraploidy. Tetraploidy was associated with response to radiotherapy when compared to diploid tumors (p = 0.07). This relationship alone did not explain the better survival of patients with tetraploid tumors, and other factors concerned with "tumor aggressivity" may be additional explanatory parameters. DNA ploidy values provide valuable information in the pretreatment situation concerning the prognosis for patients with T2/T3 bladder carcinoma.

Enhanced antitumour effect of photodynamic therapy by microtubule inhibitors.

The combination of photodynamic therapy (PDT) and the microtubule (MT) inhibitor, vincristine (VCR) or taxol, was studied in the CaD2 mammary tumour model in mice. Meso-tetra(di-adjacent-sulphonatophenyl) porphine (TPPS2a) was used as a photosensitizer. An enhanced antitumour effect was found when VCR, at an almost non-toxic dose (1 mg/kg1, was injected i.p. into the mice 6 h before PDT, while no such enhanced effect was observed when the same dose of VCR was given either 12 or 24 h before PDT or immediately before PDT. Furthermore, it was found that the number of mitotic cells increased 4-5-fold 6 h after the injection of VCR into the mice. VCR did not enhance the sensitivity of normal skin to PDT. Combination of PDT and taxol was also studied. The antitumour activity of PDT could be increased by taxol when the drug (35 mg/kg) was administered i.p. either 6 h prior to PDT or immediately after or before PDT. No significant enhancement in PDT efficiency was found when PDT with photofrin was combined with VCR.

Radiotherapy of T4 bladder carcinoma.

A 16% 5-year crude survival was observed in 159 irradiated patients with T4NXMO bladder carcinoma. The presence of a T4a tumour and a good performance status were important prognostic parameters. The combination of radiotherapy and weekly injections of 5-FUra (12 mg/kg) resulted in a significant 2-year survival increase. New regimens of combined radiotherapy/chemotherapy should be developed for patients with T4NXMO bladder carcinoma. The palliation effect of radiotherapy should further be evaluated, preferably in prospective studies comparing radiotherapy with other types of palliation treatment.

Prognosis in patients with metastatic non-seminomatous testicular cancer.

155 patients with metastatic non-seminomatous testicular cancer were treated with cisplatin-based chemotherapy which in most cases was combined with surgery. The 5 year crude survival was 90% for all patients (98 patients with small volume disease: 97%; 32 patients with large volume disease: 91%; 25 patients with very large volume disease: 64%). High pre-chemotherapy serum tumour marker levels (AFP greater than 500 micrograms/l; and/or HCG greater than 1000 U/l) decreased the survival rates in all groups. Only 4 of 17 relapsing patients were rendered tumour-free by salvage chemotherapy. In a multivariate analysis, a pre-chemotherapy alpha-foetoprotein (AFP) level greater than 500 micrograms/l was associated with poor survival as was the presence of a retroperitoneal tumour greater than 10 cm, lung metastases greater than 3 cm and/or extrapulmonary hematogenous metastases. It is concluded that easily assessable clinical pre-treatment variables can be used to define high risk or low risk patients with metastatic testicular cancer. Treatment intensity should be adjusted in accordance to such prognostic factors.

Nuclear feulgen DNA content and nuclear size in human breast carcinoma.

Biopsy specimens from 29 patients with operable breast carcinomas were examined by Feulgen-DNA microspectrophotometry. A diploid DNA stemline was found in ten tumors, seven of which were stage I carcinomas. In 19 tumors, ten of which were stage II cancers, a non-diploid DNA stemline was observed. The diploid tumors were most often estrogen receptor-positive (nine of nine examined carcinomas), whereas 13 of 19 non-diploid tumors studied did not contain estrogen receptors. The mean nuclear size of the non-diploid tumor cells was increased compared with that of the diploid malignant cells. Three grade I carcinomas were diploid, whereas three grade III tumors were non-diploid. Of the 23 grade II carcinomas, seven were diploid and 16 non-diploid. Hum Pathol 13:626-630, 1982.

Cooperative inactivation of cells in microcolonies treated with UVA radiation.

Microcolonies of Madison-Darby canine kidney cells (MDCK II) were exposed to UVA radiation, and the number of cells with membrane damage was determined by staining with propidium iodide and fluorescence microscopy. The cells were clearly damaged in a nonrandom manner: The distribution of damaged cells per microcolony was incompatible with the assumption that the cells were damaged independently. The data were accurately described by a so-called propagated damage model in which a damaged cell can influence its neighbors in a propagating manner. These findings do not agree with the common view that optical radiation interacts with cells in a way in which damage manifested in a cell is the result of absorption of light in the same cell.

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.

5-Aminolevulinic acid-based photochemical internalization of the immunotoxin MOC31-gelonin generates synergistic cytotoxic effects in vitro.

Photochemical internalization (PCI) is a novel method for the endosomal or lysosomal release of membrane-impermeable molecules into the cytosol of target cells. This novel technology is based on the photodynamically induced rupture of endocytic vesicles preloaded with molecules of therapeutic interest. PCI of the ribosome-inactivating plant toxin gelonin and the immunotoxin monoclonal antibody 31 (MOC31) gelonin has been performed previously by the use of the endocytic vesicle-localizing photosensitizers TPPS2a and AIPcS2a and light, demonstrating synergistic toxicity against the more than 20 different cell lines tested, most of them of neoplastic origin. In this study we demonstrate that 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) is also capable of inducing PCI of MOC31-gelonin in the human colon adenocarcinoma cell line WiDr. The cells were incubated with 1 mM 5-ALA for up to 8 h in serum-free medium and from 24 to 96 h in serum-containing medium. Fluorescence microscopical studies indicate a partial plasma membrane localization of PpIX when 5-ALA was applied under serum-free conditions. This plasma membrane localization was not seen when 5-ALA was given in the presence of serum. There was a granular component of the PpIX localization in addition to a diffuse cytoplasmic localization. The granular component resembled the localization of the fluorescent dye conjugate Alexa-gelonin and the lysosomal localizing dye acridine orange. Our present results provide evidence for an endocytic vesicle-associated fraction of PpIX after 5-ALA incubation of the WiDr cells. We demonstrate that PCI, by combining 5-ALA, MOC31-gelonin and light, induces a synergistic cytotoxic effect against the WiDr cells.