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Treatment of malignant pericardial effusion with 32P-colloid.

Malignant pericardial effusion is usually treated only when signs of cardiac tamponade develop. Several methods of treatment have been reported with an overall response rate of approximately 75%. Since our initial study using intrapericardial 32P-colloid instillation as a treatment modality for pericardial effusion demonstrated a significant higher response rate, this study was conducted to further evaluate the efficacy of intrapericardial 32P-colloid in terms of response rates and duration of remissions. Intrapericardial instillation of 185-370 MBq (5-10 mCi) 32P-colloid in 36 patients with malignant pericardial effusion resulted in a complete remission rate of 94.5% (34 patients) whereas two patients did not respond to treatment due to a foudroyant formation of pericardial fluid. The median duration time was 8 months. No side-effects were observed. These results suggest that intrapericardial instillation of 32P-colloid is a simple, reliable and safe treatment strategy for patients with malignant pericardial effusions. Therefore, since further evidence is provided that 32P-colloid is significantly more effective than external radiation or non-radioactive sclerosing agents, this treatment modality should be considered for the management of malignant pericardial effusion.

Breast Neoplasms↗

Metabolic factors affecting enhanced phosphorus uptake by activated sludge.

Activated sludges obtained from the Rilling Road plant located at San Antonio, Tex., and from the Hyperion treatment plant located at Los Angeles, Calif., have the ability to remove all of the orthophosphate normally present in Tucson sewage within 3 hr after being added to the waste water. Phosphorus removal was independent of externally supplied sources of energy and ions, since orthophosphate and (32)P radioactivity were readily removed from tap water, glass-distilled water, and deionized water. Phosphorus uptake by Rilling sludge in the laboratory appears to be wholly biological, as it has an optimum pH range (7.7 to 9.7) and an optimum temperature range (24 to 37 C). It was inhibited by HgCl(2), iodoacetic acid, p-chloromercuribenzoic acid, NaN(3), and 2, 4-dinitrophenol (compounds that affect bacterial membrane permeability, sulfhydryl enzymes, and adenosine triphosphate synthesis). Uptake was inhibited by 1% NaCl but was not affected by 10(-3)m ethylenediaminetetraacetic acid disodium salt (a chelating agent for many metallic ions).

Arizona↗

Effects of a tumor promoter on phospholipid metabolism in HeLa cells.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) caused a marked stimulation of inorganic [32P]orthophosphate incorporation into HeLa-cell phosphatidylcholine (PC), phosphatidylethanolamine (PE), and lysophosphatidylethanolamine. The increased incorporation of inorganic [32P]orthophosphate into PE and lysophosphatidylethanolamine in the presence of TPA was not associated with an increase in PE synthesis as detected by the incorporation of [3H]serine or [3H]ethanolamine. The PC-specific exchange protein from beef liver was used to insert PC labeled with [3H]choline, inorganic [32P]orthophosphate, or [14C]arachidonic acid plus [3H]palmitic acid into the outer monolayer of intact HeLa cell membranes. Radioactivity from the latter two compounds was rapidly incorporated into PE and lysophosphatidylethanolamine; the incorporation was stimulated by TPA. It was concluded that TPA stimulated the formation of PE by base exchange between ethanolamine and PC.

HeLa Cells↗

[Pharmacokinetics of P32-phosphemide in the body of rats with sarcoma 45].

Pharmacokinetics of phosphamide-32P was studied in experiments on rats with sarcoma 45 and intact (tumourless) ones. Soon (15 min to 1 hour) after intraperitoneal injection of the compound the maximum amount of the isotope was definable in the liver, kidneys, spleen, suprarenals, in the tumour and the thymus. The content of the compound in the tumour ammounted to 0.2--0.4 per cent of the radioactivity introduced. Distribution of phosphemide-32P among organs and tissues of the rats with sarcoma 45 quatitatively differs from that in intact animals. During the first 24 hours 55--56 per cent of the radioactivity introduced is excreted together with urine.

Adrenal Glands↗

Disposal of spent tributylphosphate by gliding arc plasma.

The gliding arc in humid air is a relevant source of free radicals and strongly oxidising species such as HO* (shown by emission spectroscopy), which are able to degrade organic wastes. This feature was used in a new process for mineralising spent tributylphosphate (TBP) which is an important waste from nuclear industry. The degradation kinetics is examined by monitoring the conversion of TBP into phosphoric acid in a batch reactor. The kinetics exhibits three steps and especially an overall zero-order linear step with a rate of 10 mmol h(-1) at the beginning of the treatment. This zero-order step agrees with a surface oxidation process. After a 13.7h treatment, about 45% of the TBP is converted into inorganic phosphorus compounds, with phosphoric acid as the major product (63% of inorganic phosphorus compounds), and at least 19.5% is not degraded. Dibutylphosphoric acid (HDBP) was identified as the main by-product by a nuclear magnetic resonance technique, infrared spectroscopy and gas chromatography.

Free Radicals↗

[Calcium absorption by the rat from various milks in relation to their total phosphorus and lactose contents].

The absorption of calcium corresponds to a strictly determined mechanism inhibited by phosphates and activited by carbohydrates. We investigate in what extent the absorption of milk calcium from various species has the same proceeding. 4 months old rats are given orally solutions of CaCl2 alone or in combination with sodium dihydrogen phosphate or lactose or these both compounds. We compare calcium absorption of these solutions to that of milk from woman, cow or sow, or to dilutions of these two latter milks. All these fluids are dosed so that they correspond each other by their respective content in total calcium, total phosphorus and lactose. Each solution contains 45Ca. Blood samples 1,30, 4, 6 and 24 hours after ingestion allow establishing the variations of plasma radioactivity. Rats are sacrified after 24 hours. In certain cases, samples from digestive tube contents and feces provide a coefficient of calcium absorption. The osseous retention is obtained from femur radioactivity. In breif, we may consider that milks from cow and sow provide calcium the absorption of which is settled after their respective content of total phosphorus and lactose. Opposed effects of these inhibitors and activators of calcium absorption compensate so that calcium from these milks is just a little better utilized by adult rat than calcium from an isocalcic solution of CaCl2. Calcium from woman milk, rich in carbohydrates, poor in total phosphorus is better absorbed than that from the two latters, however less than we might expect from its high lactose content. We may wonder that calcium utilization from cow milk is as moderate as that of a solution of CaCl2. But skeleton mineralization which may be fulfilled by milk is made better because of that: it is not chloride but calcium phosphate which ensures this mineralization, a phosphate which alone would impair this mineralization without the compensative role of lactose.

Animals↗

32P-postlabeling detection of DNA adducts in fish from chemically contaminated waterways.

Fish were collected from sites in the chemically-contaminated Buffalo River, New York, and the Detroit River, Michigan. The sediments of these rivers have high levels of chemical contaminants, including polycyclic aromatic hydrocarbons (PAHs), and fish from these locations have high prevalences of liver cancer. To determine chemical-DNA interactions and a possible role for chemicals as a cause of the observed tumors, DNA was isolated from livers and was enzymatically digested to normal and adducted nucleotides. The DNA digests were enriched for hydrophobic, bulky adducts, either by preparative reverse phase high pressure liquid chromatography, or by selective nuclease P1 dephosphorylation of normal nucleotides. DNA-chemical adducts were then quantitated by 32P-postlabeling analysis. Regardless of the adduct enrichment procedure, the chromatograms derived from DNA of fish from polluted areas showed a diffuse, diagonal radioactive zone consisting, at least in part, of multiple overlapping discrete adduct spots. The behavior of the adducts in the diagonal radioactive zone and of their unlabeled precursors is consistent with their identification as nucleotide adducts of a variety of bulky, hydrophobic, aromatic genotoxic compounds. Analysis of bile demonstrated recent exposure to multi-ringed aromatic compounds.

Animals↗

Stimulation of phosphate uptake in human platelets by thrombin and collagen. Changes in specific 32P labeling of metabolic ATP and polyphosphoinositides.

The uptake of [32P]phosphate by human, gel-filtered blood platelets and its incorporation into cytoplasmic ATP and polyphosphoinositides was studied. In unstimulated platelets, uptake was Na+o-dependent and saturable at approximately 20 nmol/min/10(11) cells with a half-maximal rate at 0.5 mM extracellular phosphate. Upon stimulation with thrombin or collagen, net influx of [32P]Pi was accelerated 5- to 10-fold. With thrombin, [32P]Pi efflux was also increased. After the first 2 min, efflux exceeded influx, resulting in the net release of [32P]Pi from the platelets. Since the stimulus-induced burst in [32P]Pi uptake paralleled the secretory responses, it might be an integral part of stimulus-response coupling in platelets. The stimulus-induced burst in net [32P]Pi uptake led to an enhanced labeling of metabolic ATP, which was already detectable at 5 s after stimulation with thrombin. Concomitantly, the incorporation of [32P]Pi into phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate was accelerated. The thrombin-induced increase in specific 32P radioactivity of cytoplasmic ATP fully accounted for the simultaneous increase in specific 32P radioactivity of these phosphoinositides. In studying the extent of 32P labeling of phosphorylated compounds in response to a cellular stimulus, it is therefore essential to measure the effect of the stimulus on the specific radioactivity of cytoplasmic ATP.

Adenosine Triphosphate↗

Synoviorthesis with 32P-colloidal chromic phosphate in rheumatoid arthritis--clinical, histopathologic and arthrographic changes.

Synoviorthesis was performed in 217 joints from 111 patients suffering from different stages of rheumatoid arthritis (RA). 32P-colloidal chromic phosphate was employed, with an average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as the MCP or PIP joints. Satisfactory clinical results were observed in 84% of the cases and no significant side effects resulted after a follow-up period from 1 to 10 years. Striking effects after treatment were observed through histopathological studies (light and electron microscopy) and the use of contrast arthrography. We concluded that radioactive synovectomy with 32P-chromate is a very useful method for the local treatment of RA.

Arthritis, Rheumatoid↗

Butanol extracts from myelin fragments-II. Some properties of 5-hydroxytryptamine binding.

The myelin fragments of rat brain stem were treated with butanol-water mixtures, and the butanol extracts (total extract=TE) were incubated with 5 x10(-7) M of C14.5-HT. After incubation, protein, lipid phosphorus and radioactivity were analyzed by Sephadex LH20 column chromatography. Two peaks of components eluted in chloroform-methanol 4:1 (peak I and II) showed the binding capacity for C14.5-HT. The displacement studies with unlabeled 5-HT (5 X 10(-4) M) suggested that peak II was the saturable binding component to 5-HT. On the other hand, butanol extracts from the synaptic membranes of rat brain stem did not show binding for C14.5-HT. Various compounds were studied to determine their inhibitory effects on the saturable binding of 5-HT to TE. The results indicated that acetylcholine, dopamine and tryptamine inhibited the 5-HT binding but LSD, reserpine, colchicine, vinblastine, 5-hydroxytryptophan (5-HTP) and 5-hydroxy-3-indole acetic acid (5-HIAA) had no effect.

Acetylcholine↗

Effects of two fluoride gels on fluoride uptake and phosphorus loss during artificial caries formation.

Blocks of human enamel were cycled through a demineralization--F-treatment-remineralization procedure and then analyzed for fluoride and the presence of caries-like lesions. Treatments with a sodium fluoride gel (5000 ppm F) increased the enamel fluoride concentration to 6500 ppm F, whereas a stannous fluoride gel (1000 ppm F) increased enamel fluoride to about 1200 ppm F. Although a control treatment (water) allowed caries-like lesions to form, as observed by microradiography, no lesions were found in either of the fluoride-treated groups. When the experiment was repeated with radioactive teeth, mineral loss, as determined by release of 32P, was again greatest in the water-treated control group, but some loss was observed in the fluoride treatment groups. The least loss was found in the sodium fluoride group. It was concluded that the fluoride treatments not only increased enamel resistance but also enhanced remineralization so that calcium phosphate was replaced during the subsequent remineralization phase. Because of the probability that stannous ions were deposited during the stannous fluoride treatments, some of the apparent calcium phosphate re-deposition in this group was probably stannous compounds.

Absorptiometry, Photon↗

Synthesis of fluorescently labeled alkylated DNA adduct standards and separation by capillary electrophoresis.

DNA adducts are regarded as individual internal dosimeters for the exposure to chemical carcinogens. To date, the most sensitive method for DNA adduct analysis is the radioactive 32P-postlabeling method, which allows the detection of one adduct in 10(10) unmodified nucleotides in microg amounts of DNA. However, this technique suffers from disadvantages such as working with radioactive phosphorus and time-consuming chromatographic separation procedures. In addition, the simultaneous detection of adducts from different classes of carcinogens in a DNA sample is difficult. In order to overcome these drawbacks, we are developing a new detection method, comprising fluorescence labeling of DNA adducts, capillary electrophoretic (CE) separation, and on-line detection by monitoring laser-induced fluorescence (LIF). So far, we have evaluated the separation power and the detection limit of CE with fluorescently labeled standard compounds such as unmodified nucleotides or alkylated thymidines. For this purpose, we developed a universal method for labeling 5'-OH-mononucleosid-3'-dicyanoethyl-phosphates with fluorescent dyes based on the phosphoramidite technology for DNA synthesis. The separation of N3-methylated, N3-, O2- and O4-butylated thymidines from the unmodified nucleotide within a few minutes recommends CE-LIF as a powerful method for DNA adduct analysis.

Alkylation↗

Determination of levels of glycolytic intermediates and nucleotides in platelets by pulse-labeling with [32P]orthophosphate.

A method is presented for the separation and quantification of 32P-labeled carbohydrates and nucleotides in blood platelets which have been pulse-labeled with [32P]orthophosphate. The procedure is based on two-dimensional paper chromatography, identification of the spots by radioautography and enzymatic methods, and quantitation of 32P radioactivity by liquid scintillation counting. The data show that 32P is homogeneously distributed among the compounds studied so that the total radioactivity is proportional to the levels of these compounds in the metabolic compartment of the cells. Thus, this method provides a sensitive and accurate means to evaluate phosphorylated intermediates in glycolysis and nucleotide metabolism and to assess the transfer of energy-rich phosphate groups between these pathways in particular.

Autoradiography↗

Covalent modification of G-proteins by affinity labeling.

The activation of heterotrimeric G-proteins is tightly regulated by the exchange of GTP for GDP in the alpha-subunit; mostly--but not exclusively--seven-transmembrane receptors function as the guanine nucleotide exchange factors (GEFs). A research goal may be to determine which G-protein alpha-subunit is activated by the receptor under investigation. In a membrane preparation obtained from cells or tissues this can be achieved in a seemingly straightforward manner by determining if the receptor increases the covalent incorporation of GTP analogs into G-protein alpha-subunits. Because the GTP analogs may be labeled to high specific radioactivity the alpha-subunit can then be identified with the use of specific antibodies. One of the compounds we present here (2',3'-dialdehyde-GTP) can also be employed to block receptor-mediated G-protein activation and to disrupt the cognate signaling pathway.

Affinity Labels↗

Intraperitoneal chromic phosphate P 32 suspension therapy of malignant peritoneal cytology in endometrial carcinoma.

Malignant peritoneal cytology in patients with endometrial carcinoma is a poor prognostic feature, identifying patients at high risk for early intra-abdominal recurrence. Between 1977 and January, 1983, 65 women with endometrial carcinoma who had malignant peritoneal cytology were treated with adjuvant intraperitoneal radioactive chromic phosphate P 32 suspension. Fifty-three patients (80%) were clinical Stage I, nine (14%) were Stage II, and three (7%) were clinical Stage III. Life-table estimates of disease-free survival were 89% for clinical Stage I patients and 94% for surgical Stage I patients beyond 24 months. One patient developed an intraperitoneal recurrence, four had simultaneous intraperitoneal and extraperitoneal recurrences, and six developed recurrences outside of the peritoneal cavity. Few significant acute complications occurred after therapy with radioactive chromic phosphate P 32 suspension. Chronic intestinal morbidity that required surgical correction was encountered in five of 17 patients (29%) who received adjuvant pelvic radiation, compared to none of the 48 patients (0%) who received only radioactive chromic phosphate P 32 suspension (p less than 0.001). Intraperitoneal instillation of radioactive chromic phosphate P 32 suspension is effective therapy for patients with malignant peritoneal cytology from endometrial carcinoma. Caution should be exercised when radioactive chromic phosphate P 32 suspension and external radiation therapy are combined.

Adenocarcinoma↗

The acute effect of sodium cellulose phosphate on intestinal absorption and urinary excretion of calcium in man.

Intestinal 47Ca absorption has been determined from blood and stool radioactivity after oral administration of the isotope in nine patients before and during cellulose phosphate treatment. Oral administration of 5 g cellulose phosphate concomitant with 47Ca in 100 mg "carrier" calcium as CaCl2 decreased 47Ca absorption by 80 per cent. Cellulose phosphate, 5 g three times daily, decreased urinary excretion of non-radioactive calcium by 47 percent. Urinary magnesium excretion decreased by 47 percent whereas urinary phosphorus excretion increased by 67%. Calcium and magnesium excretion in urine decrease because cellulose phosphate binds divalent cations within the GI tract. The increased phosphorus excretion is probably due to partial hydrolysis of the substance in the gut.

Calcium↗