99mTc-ENS: a new radiopharmaceutical for aerosol lung scintigraphy. Comparison between different freeze-dried formulations.

UNLABELLED: Exogenous natural surfactant (ENS) labeled with 99mTc(99mTc-ENS) is a new radiopharmaceutical for pulmonary aerosol scintigraphy. In this study, different freeze-dried formulations were evaluated to develop a suitable and long-storage method for the ENS, the nonradioactive precursor of this radiopharmaceutical. METHODS: Two freeze-dried formulations were evaluated: the sterile ENS suspension-stannous chloride altogether lyophilized (chlorlioENS) and the lyophilized sterile ENS suspension with the addition of stannous chloride as a solid drug (lioENS). These precursors were stored at room temperature for 3 mo and then labeled with 99mTc. For comparative purposes, the sterile ENS suspension with the addition of stannous chloride labeled with 99mTc(99mTc-chlorENS) was also studied. The quality controls for each radiopharmaceutical were performed by an ascending paper chromatography to determine the labeling yield percentages. The study was performed in 30 female Sprague Dawley rats, which inhaled each radiopharmaceutical by nebulization. Twenty-five minutes after the aerosol inhalation, the animals were killed to extract their organs and measure their activity in a gamma spectrometer. The data are given as the percentage of activity concentration (C%) for each organ. RESULTS: The physicochemical properties of lioENS were adequate for a freeze-dried product. The labeling yields for 99mTc-lioENS and for 99mTc-chlorENS were always greater than 95% even after nebulization. The results of the biologic distribution studies showed that the activity concentration found in lungs for these radiopharmaceuticals were 95.7% +/- 2.6% and 96.7% +/- 2.6% respectively, results that do not differ statistically. On the other hand, the activity concentration found in lungs for the 99mTc-chlorlioENS (31.3% +/- 11.1%) and its labeling yield percentages (<10%) are statistically different (P < 0.05) from the results obtained with the two radiopharmaceuticals mentioned above. CONCLUSION: Taking into account the lioENS physicochemical properties, its long shelf life and that 99mTc-lioENS shows the same radiochemical and radiopharmacological behavior of the 99mTc-chlorENS, it can be concluded that the 99mTc-lioENS can be used for aerosol lung scintigraphy.

A new procedure to fortify fluid milk and dairy products with high-bioavailable ferrous sulfate.

The behavior of SFE-171 used for food fortification was studied. The biological and nutritional properties of this new iron source are discussed in this work.

Use of colloids of chromic [32P] phosphate in treatment of solid tumors.

In order to evaluate the effectiveness of an intratumorally single dose of chromic [32P] phosphate for the treatment of solid tumors, studies of bioelimination, biodistribution, and therapeutic action were carried out. Only for comparative purposes were similar studies undertaken using a solution of sodium [32P] orthophosphate-gelatin. Results show that when sodium [32P] orthophosphate-gelatin was intratumorally injected, the percentage of total elimination, after 32 days of treatment, was equal to 85.90 +/- 8.70%, with a higher percentage in urine (64.50 +/- 13.70%) than in feces (21.40 +/- 4.50%). In biodistribution studies, the greater percentage was found in bone (15.54 +/- 2.21%), whereas only 2.51 +/- 0.39% remained in the tumor. When chromic [32P] phosphate was intratumorally injected, we found that the total elimination was equal to 51.70 +/- 6.90%, with a higher amount in feces (32.70 +/- 4.80%) than in urine (19.00 +/- 3.60%). Biodistribution studies demonstrated that 28.93 +/- 1.30% was still in the tumor and 19.01 +/- 1.30% of the injected activity was found in the liver. On the other hand, when therapeutic action was evaluated, no tumoral regression was observed. These results demonstrate that the colloid of chromic [32P] phosphate cannot be used in the treatment of solid tumors as it mobilizes from the injection point, delivering a high dose to the entire organism.

Radiochemical and radiopharmacological properties of pirocarbotrat and other labeled charcoal dispersions: comparative studies in rats with NMU-induced mammary tumors.

The purpose of this work is to study the physicochemical properties of Pirocarbotrat to explain its radiopharmacological behavior. We also studied a mixture of charcoal plus chromic [32P]phosphate and charcoal plus sodium [32P]orthophosphate only for comparative purposes. The results show that the mean diameter of the Pirocarbotrat particles was 2.5 microm with an homogeneous distribution, while the other products show an heterogeneous distribution of the particle sizes, with a mean size diameter between 0.5 and 0.9 microm. Hydrolysis studies with a solution of 0.1 N HCl and with sulfochromic mixture revealed that in Pirocarbotrat the 32P is strongly bound to the charcoal particles. Bioelimination studies of Pirocarbotrat show that the total eliminated activity was 12.70 +/- 3.90%, with a higher amount in urine (8.30 +/- 1.80%) than in feces (4.40 +/- 3.50%). When biodistribution studies of Pirocarbotrat were carried out, we found that the 84.50 +/- 2.60% of the activity remained in the tumor with almost null irradiation of the other organs under study. When therapeutic action was evaluated, we observed that the percentage of tumor regression was 78.3% for the tumors injected with Pirocarbotrat. The other dispersions under study showed different behaviors with high activity percentages distributed throughout the organism. These studies demonstrate that Pirocarbotrat has the best radiopharmacological properties to ensure irradiation of the tumor with the least concomitant irradiation of surroundings or other organs or tissues.

Pirocarbotrat: a new radiopharmaceutical for the treatment of solid tumors--comparative studies in N-nitrosomethylurea-induced rat mammary tumors.

To evaluate the effectiveness of a single intratumoral dose of Pirocarbotrat, a gelatin-protected charcoal suspension labeled with chromic [32P]pyrophosphate, studies of bioelimination, biodistribution and therapeutic action were carried out in rats, and the results obtained were compared with those of other 32P dispersions. We found that 78.3% of the treated tumors reduced size after 32 days of treatment. At that time, the total eliminated activity was 12.70 +/- 3.90% distributed in urine (8.30 +/- 1.80%) and feces (4.40 +/- 3.50%). Biodistribution studies demonstrate that 84.50 +/- 2.60% of the injected activity remained in the tumor, with no significant concentration in the rest of the organism. We conclude that Pirocarbotrat can be used as a safe agent for brachytherapy of solid tumors with beta particles.

Further studies on the histamine metabolism in the M-2 adenocarcinoma.

Further studies of the histamine metabolism in the M-2 adenocarcinoma have shown that the tumor and the intestinal histidine decarboxylase should have similar conformations, since both enzymes exhibit superimposable curves of activity vs. pH as well as nearly identical 50% inhibitory concentrations for the specific inhibitor NSD-1015. It has also be found that the increase of the radioactivity uptake by the tumor nuclear fraction compared to the intestinal nuclear fraction after their incubation with 3H-histamine, may be due to qualitative differences between the nuclei, since endogenous contents of DNA and associated proteins were not significantly different. The specific binding of 3H-histamine to different purified DNAs demonstrated very dissimilar proportions of bound histamine, suggesting a possible interaction of histamine with DNA, similar to some of the proposed mechanisms for the modulation of the genomic expression of DNA by some polyamines.

Histamine receptors in an experimental mammary carcinoma.

An experimental mammary carcinoma was induced in Sprague-Dawley rats by the ip administration of N-nitroso-N-methylurea (NMU) in three doses of 50 mg/kg. In order to study the expression of histamine receptors in these experimental tumors, the presence of specific binding sites for histamine was studied. Using [3H]-histamine as a radioligand, two specific binding sites were characterized on the cell membrane. The first site, of high affinity, Kd = 4 +/- 2 nM, was further characterized as an H2 type using [3H]-cimetidine and [3H]-tiotidine as radioligands and by displacement experiments with different histamine agonists and antagonists. The second one of low affinity, Kd = 35 +/- 14 nM, needs further characterization. The determination of cAMP levels showed that histamine and the H2 agonist dimaprit, produced a significant decrease in the nucleotide concentration 6 minutes after stimulation, in a response that was specifically abolished by H2 antagonists. Based on these results, we conclude that neoplastic cells from NMU induced tumors express H2 histamine membrane receptors which are coupled to a transductional pathway different from cAMP production, which may be involved in the regulation of tumor growth.

Bioavailability, biodistribution, and toxicity of BioZn-AAS(1): a new zinc source. comparative studies in rats.

Food fortification with a proper zinc compound is an economic and effective strategy to prevent zinc deficiency. BioZn-AAS, a zinc gluconate stabilized with glycine, was compared with zinc sulfate (reference standard), zinc hydroxide, and zinc gluconate, all of them labeled with (65)Zn. This preclinical study was performed on Sprague-Dawley rats of both sexes, and the administered dose was 85 microg/kg of zinc. Bioavailability studies showed that absorption of BioZn-AAS was not statistically different than absorption from other sources in female rats (25.65% +/- 2.20% for BioZn-AAS, 28.24% +/- 4. 60% for ZnSO(4), 24.91% +/- 4.02% for Zn[OH](2), and 25.51% +/- 2. 70% for Zn-gluconate). In the case of the male rats, absorption of BioZn-AAS (27.97% +/- 4.20%) was higher (P<0.05) than that from the other compounds (23.15% +/- 2.90% for ZnSO(4), 22.62% +/- 3.90% for Zn[OH](2), and 22.30% +/- 3.90% for Zn-gluconate). Biodistribution studies demonstrated that the zinc from BioZn-AAS followed the same metabolic pathway as zinc from the other sources. Toxicity studies were performed with 50 female and 50 male rats. The value of oral lethal dose 50 (LD(50)) was 2000 mg/kg for female rats and 1900 mg/kg for male rats. Therefore, we conclude that BioZn-AAS has adequate properties to be considered a proper zinc compound for food fortification or dietary supplementation.

Bioavailability study of dried microencapsulated ferrous sulfate--SFE 171--by means of the prophylactic-preventive method.

Microencapsulated ferrous sulfate with soy lecithin (SFE-171) has been used as an iron source for the fortification of milk and dairy products. With the purpose to extend the use of this agent to other kind of foods or even to pharmaceutical preparations for oral administration, the SFE-171 was turned into a fluid powder (SFE-171-P) by means of vacuum drying. The iron bioavailability (BioFe) of SFE-171-P was evaluated in this work by means of the prophylactic-preventive method in rats, using ferrous sulfate as reference standard. Both iron sources were separately added to a basal diet of low iron content in a concentration of 10 mg iron/kg diet. Two groups of 10 weaned rats 25 days old received the fortified diets during 28 days, while a third group of the same size received the basal diet without iron additions. The weights and haemoglobin concentrations (HbC) of every animal were determined before and after the treatment, thus allowing the calculation of the mass of iron incorporated into haemoglobin (HbFe) during this period. The BioFe of the iron sources were obtained as the percentage ratio between the HbFe and the mass of iron consumed by each animal. The results were also given as Relative Biological Value (RBV), which relates the BioFe of the studied source with that of the reference standard. The liver iron concentration (LIC) of each animal was determined at the end of the experiment in order to evaLuate the influence of the studied iron sources on the liver iron stores. SFE-171-P presented BioFe, RBV and LIC values of (47 +/- 7)%, 109% and (46.6 +/- 3.4) mg/kg respectively, while the corresponding values for the reference standard were of (43 +/- 7)%, 100% and (45.0 +/- 4.7) mg/kg. These results show that the drying process used to produce the SFE-171-P does not affect its bioavailability, which is also adequate for the potential use of this product in food fortification or with pharmaceutical purposes.

Bioavailability, absorption mechanism, and toxicity of microencapsulated iron (I) sulfate: studies in mice.

The iron compounds used for food fortification have to meet certain requisites related to their bioavailability, absorption mechanism, and toxicity, since they will be consumed by a massive population group. With these purposes, we evaluated a new product used for the iron fortification of milk and lacteous derivatives, called SFE-171, which is a ferrous sulfate, microencapsulated with phospholipids. The bioavailability studies were carried out using four groups of 30 female mice each. In two groups, we studied the absorption of ferrous ascorbate and ferrous sulfate, both in water as reference standards, which show absorptions of 13.1+/-4.9% and 13.2+/-4.3%, respectively. With the third group, we studied the absorption of ferrous sulfate in milk; its value, 7.9+/-3.2%, is significantly lower than that of the remaining groups, with a p < 0.01. The studies with SFE-171 in milk, were performed on the fourth group, with a result of 11.6+/-4.5%, demonstrating that its absorption does not differ significantly from that of the reference standards. The absorption mechanism was determined by means of in vivo self-displacement studies of the ferrous ion and the SFE-171, taking ferrous sulfate as the reference compound. For this study, 210 female mice were used, and no significant difference between the absorption mechanism of both products could be observed. Toxicity studies of the new product with regard to ferrous sulfate were carried out with two groups of 70 female mice each and two groups of 70 male mice each. The lethal dose 50% LD50 for SFE-171 and for ferrous sulfate was 1200 and 680 mg/kg for female mice and 1230 and 670 mg/kg for male mice, respectively, demonstrating that the toxicity of the first product is substantially lower than that of the reference standard. We conclude that the iron product under study has a high bioavailability, an absorption mechanism equal to that of nonhemic iron, and lower toxicity than ferrous sulfate.

Microencapsulated ferrous sulfate to fortify cow milk: absorption and distribution in mice.

To determine the absorption and biodistribution of iron from microencapsulated ferrous sulfate (SFE-171), used to fortify dairy products with iron, a comparative study in four groups of 30 mice each was carried out. In two of the groups, the absorption of iron from ferrous ascorbate in water (13.3 +/- 4.3%) and from ferrous sulfate in water (12.7 +/- 3.9%) was determined and taken as reference standards. In the third group the iron absorption from SFE-171 in milk was determined, giving a value of 12.1 +/- 4.2%, which statistically does not differ from the data obtained with either reference standard. In the fourth group, the absorption of iron from ferrous sulfate in milk showed a value of 7.7 +/- 3.4%, which statistically differs with a p < 0.01 from the data corresponding to the other three groups. The biodistribution studies showed that the iron from the SFE-171 follows the same metabolic pathway as the iron from the reference standards thus, giving a higher radioactivity percentage and radioactivity concentration in organs or systems, principally blood, that are closely related to iron metabolism. Our studies allow us to conclude that the iron from SFE-171 in milk follows the same behavior as the nonhemic iron, with a higher absorption than that of ferrous sulfate in milk.

Bioavailability and stability of microencapsulated ferrous sulfate in fluid milk: studies in mice.

Iron deficiency is the most important nutritional problem all over the world. Fluid milk is an attractive vehicle for iron fortification, since it is a food with a high nutritional value, accessible to the whole population and easy to be given to children. Fortification of this food with iron has the disadvantage of the interaction of the iron with the constitutive elements of milk, diminishing its bioavailability and changing its sensorial properties, making it unacceptable. Nowadays, this problem can be overcome by the implementation of a new technological procedure, which consists in the microencapsulation of the ferrous sulfate with lecithin, thus avoiding the interaction of iron with the food. The absorption obtained in mice for milk-ferrous sulfate was 7.9 +/- 3.2%, while for microencapsulated ferrous sulfate-milk the result was 11.6 +/- 4.5%. Comparing these data with those obtained with the ferrous ascorbate in water 13.1 +/- 4.9% and ferrous sulfate in water 13.2 +/- 4.3%, both of them considered as reference standards, no statistically significant difference between them and the microencapsulated ferrous sulfate in milk can be observed. However, this difference becomes significant (p < 0.01) when these products are compared to the non-encapsulated ferrous sulfate in milk. On the other hand, we demonstrated that this product is stable to heat-processing (100 degrees C, 30 min) and storage at a room temperature up to 6 months that lacteous products are usually submitted to.

Study of industrial microencapsulated ferrous sulfate by means of the prophylactic-preventive method to determine its bioavailability.

Radio-iron tests are frequently used to measure the bioavailability of different iron sources for food fortification. As the labeling procedures must be done under laboratory conditions, complementary studies should be carried out to evaluate the bioavailability of iron sources produced on an industrial scale. The iron bioavailability of SFE-171 (ferrous sulfate microencapsulated with phospholipids) was studied in previous reports using the compounds labeled with 59Fe and 55Fe; the results showed an iron bioavailability similar to that of ferrous sulfate. In the present work, the iron bioavailability of industrial SFE-171 was studied by the prophylactic-preventive method in rats using ferrous sulfate as the reference standard. Elemental iron powder was also studied by the same method for comparative purposes. The liver iron concentration of each animal was determined at the end of the experiment in order to evaluate the influence of each iron source on the liver iron stores. Relative biological values of 98 and 34% were found for SFE-171 and elemental iron powder, respectively, while the corresponding relative liver iron concentrations were 104 and 45%. The results provided by the prophylactic-preventive method show that the iron bioavailability of industrial SFE-171 is similar to that of ferrous sulfate; these results are also in agreement to those obtained with the radioactive compounds. We can conclude that the SFE-171 obtained by industrial procedures for massive use in iron food fortification has the same bioavailability as that of the SFE-171 produced and labeled under laboratory conditions.

Zinc and diabetes mellitus: is there a need of zinc supplementation in diabetes mellitus patients?

Diabetes mellitus is a group of metabolic disorders, the incidence of which varies widely throughout the world. The treatment of diabetes mellitus includes insulin, oral antidiabetic agents, and dietary regimens. Although the emphasis is on macronutrients intakes, there is strong evidence that there is an abnormal metabolism of several micronutrients in diabetic individuals. Zinc is one of the essential micronutrients of which status and metabolism is altered in this condition. This work is a short review about the close relation among zinc, glucose metabolism, and insulin physiology, as well as about the few experimental data about zinc absorption and zinc supplementation in diabetes mellitus patients.

New procedure to fortify fluid milk and derivatives with iron: a comparative study in mice.

It is known that the fortification of milk and its derivatives with iron has been recognized as a worldwide necessity, since the deficiency of this element produces different metabolic disorders. With this purpose, we have studied the iron absorption in mice after the administration of fluid milk and yogurt with FeSO4, stabilized by microencapsulation with soy lecithin (SFE-171) or with FeSO4, both labeled with 59Fe, which was used in the same experimental conditions for comparative purposes. The absorption of iron is influenced by the presence of some additives which usually are ingested together with milk, such as cacao, coffee, tea, "Argentine green herbs tea or maté" or cereals. The experimental results demonstrate that the iron absorption from SFE-171 is (12.3 +/- 2.9)%, whereas that from FeSO4 is (7.7 +/- 2.7)%, this difference is highly significant at p < 0.01. In all the cases, groups of 25 mice each were used to increase the statistical robustness of the experimental results. On the other hand, it could be demonstrated that the presence of 10% w/v cacao (Nesquik) increases the iron absorption in both cases, whereas 1% w/v cacao (Nesquik) and maté has no influence on the iron absorption if SFE-171 is used. Cereals (Nestum 3 cereals), yogurt and other additives like tea and coffee, for different reasons, decrease the absorption of this element.

Modification of the ferrozine technique to analyze iron contents in different foods: comparative study using an internal standard as reference methodology.

A methodology for the determination of iron in foods fortified with this element or in nutritional products is important and has to be sensitive and rapid. In developing countries, an inexpensive and reliable methodology is also required. For this purpose, the Gordon's Ferrozine technique was slightly modified and assayed with yogurt, dry powdered milk, and cereal mixtures, all of them fortified with iron, using an internal standard as the reference methodology. The obtained results demonstrate a close correlation between the standard curve interpolation method and the internal standard reference method (correlation coefficient r2 = 0.9950) in a wide range of concentrations. The slope (0.9998+/-0.0040) demonstrates that both procedures measure equal amounts of iron. The conclusion is that the proposed technique is a reliable, practical, and inexpensive methodology for iron determination in different foods fortified with iron.

Biosynthesis and subcellular localization of histamine in H-110 lymphoma.

The histidine decarboxylase activity has been studied in the H-110 Lymphoma implanted in Balb-c mice. The enzymatic activity has been determined using DL-Histidine 1-14C by measuring the I--14CO2 liberated during the incubation. It has been found that the histidine decarboxylase activity in the tumor is the highest of the studied tissues. The pretreatment for 5 days with two cytostatic drugs, cyclophosphamide and 5-fluoruracile, determined a decrease of the enzymatic activity in the tumor of 33 and 54 per cent, respectively. This effect has not been observed in the lung. The pretreatment of the animals during 10 days with 0.1 mg per kg body of histamine, which in other experimental tumors induces an increase of the endogenous content of histamine, produces a decrease of the tumoral histidine decarboxylase activity. The histamine-14C uptake was not modified by this pretreatment. The subcellular localization of the radioactivity after pretreatment with histamine-14C evidenced that the nuclear fraction of the tumor contained between 2 and 10 times the radioactivity of other tissues of the same animal.

A simplified competition data analysis for radioligand specific activity determination.

Non-linear regression and two-step linear fit methods were developed to determine the actual specific activity of 125I-ovine prolactin by radioreceptor self-displacement analysis. The experimental results obtained by the different methods are superposable. The non-linear regression method is considered to be the most adequate procedure to calculate the specific activity, but if its software is not available, the other described methods are also suitable.

Great particles [32P]chromic phosphate for treatment of solid tumors.

With the purpose of studying the effectivity of an intratumoral single dose of chromic [32P] phosphate with great particles for the treatment of solid tumors, studies of bioelimination, biodistribution and therapeutic action were carried out. Only for comparative purposes, similar studies were undertaken using a solution of sodium -32P- orthophosphate-gelatine. The results show that when sodium [32P] orthophosphate-gelatine is used, the percentage of total elimination is (85.90 +/- 8.70)% with a higher percentage in urine (64.50 +/- 13.70)% than in faeces (21.40 +/- 4.50)%. In biodistribution studies, the greater percentage is found in bone (15.54 +/- 2.21)% while only a (2.51 +/- 0.39)% remains in the tumor. When great particles chromic [32P]phosphate was intratumorally injected, we determined that the total elimination is equal (36.28 +/- 6.27)%, finding a higher amount in faeces (29.44 +/- 5.26)% than in urine (6.84 +/- 2.21)%. Biodistribution studies demonstrated that (49.82 +/- 5.41)% remains in the tumor and (9.63 +/- 4.89)% of the injected activity is found in the liver. On the other hand, when therapeutic action was evaluated, we observed that the percentage of tumor regression (P.T.R.) is 52.0% for the tumors injected with chromic [32P]phosphate and 0.0% for those injected with sodium [32P]orthophosphate-gelatine. These results show that the great particles colloid of chromic [32P]phosphate is not safe enough for the treatment of solid tumors, since it is mobilized from the injection point, delivering a high dose to the whole organism.

New methodology for pancreatic scintiscanning imaging using 125I-L-3 Iodo-alpha-Methyl Tyrosine.

The difficulty of a reliable diagnosis of pancreatic diseases by scintiscanning, is mainly derived from the lack of adequate radiopharmaceuticals. With this purpose 125I-L-3 Iodo-a-Methyl Tyrosine (125I-IMT) has been studied, which has also been used for the diagnosis of different kind of brain tumors. The purpose of this work is the development of a quick and easy method for the synthesis and purification of the 125I-IMT in order to be used in a Nuclear Medicine Service. The L-alpha-Methyl Tyrosine was labeled with 125I using I-/I03 and afterwards purified by an anionic exchange resin. The labeling yield obtained was (96.0 +/- 0.5)% when the incubation time was 15 minutes. No significant statistical differences were observed when the incubation time was extended to 1 hour. Biodistribution studies in mice show that the percentage of activity concentration in pancreas is (34.24 +/- 14.03)% at 15 minutes post injection, remaining constant for 30 minutes. The pancreas/liver ratio 15 minutes after the injection of the labeled product was (12.22 +/- 3.59) and it remained constant for 45 minutes more. These results show that 125I-IMT can be used as a diagnostic agent for pancreatic diseases. Since 123I was not available at the moment, this new methodology was developed with 125I.