Evaluation of different formulations to label exogenous natural surfactant labeled with (99m-Tc-ENS): stability and toxicity studies

El surfactante natural exógeno (ENS) marcado con 99mTc (99mTc-ENS) es un nuevo agente ventilatorio para la centellografía aérea pulmonar. Con el fin de elegir una formulación estable de ENS para ser marcado rutinariamente con 99mTc en Centros de Medicina Nuclear, las propiedades de dos formulaciones (ENS + cloruro estannoso + ácido gentísico y ENS + cloruro estannoso + ácido ascórbico) y una formulación control liofiolizada del ENS fueron analizadas mensualmente por un período de 12 meses. Sus propiedades fisicoquímicas, sus porcentajes de marcación y sus distribuciones biológicas fueron adecuadas durante este período. La formulación ENS + cloruro estannoso + ácido gentísico presenta la menor dispersión en los resultados de biodistribución, por lo cual esta formulación fue elegida para la futura producción de 99mTc-ENS. El estudio de toxicidad aguda de esta última formulación demostró que la dosis tóxica es al menos 1000 veces mayor que la dosis diagnóstica

Changes in reactive oxygen species (ROS) production in rat brain during global perinatal asphyxia: an ESR study.

A large body of evidence suggests that the production of reactive oxygen species (ROS) can play an important role in ischemic neuronal injury. However any studies has been performed in hypoxic conditions. In the present experiments we studied using electron spin resonance (ESR) techniques the ROS release in neostriatum of newborn rats subjected to acute perinatal asphyxia (PA) followed by various periods of reoxygenation. Pregnant rats' uteri still containing foetuses were taken out and subjected to PA by immersion in a 37 degrees C water bath during the following periods of time: 5, 10, 15, 19 and 20 min. After performing PA, animals were recovered and ROS measured after 0, 5, 15, 30 or 60 min of reoxygenation. Then, pups were sacrificed, their neostriatum removed and homogenised with N-tert.-butyl-alpha-phenylnitrone (PBN) and diethylenetriamine-pentacetic acid (DPTA) in phosphate-buffered saline (PBS) and the formed complexes were extracted with ethyl acetate an analysed using an X-band ESR spectrometer. A significant release of ROS was detected at 19 and 20 min of PA after 5 min of reoxygenation. These data provide strong evidence that ROS could be involved in neuronal damage during PA.

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.

Zinc status and immune system relationship: a review.

The essentiality of zinc for humans was first documented by Prasad in the 1960s. The main clinical manifestations associated with zinc deficiency are growth retardation, hypogonadism, diarrhea, and increased susceptibility to infectious diseases. Thus, in the past 25 yr, there was an increased interest of researchers in studying the role of zinc in human immunity. Although mechanistic research has been carried out using animal models, there are several studies in humans with similar results. This work is an attempt to review the information available in this field to understand the important role that zinc plays in the normal development and function of the immune system.

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.

99mTc-ENS ventilation scintigraphy: preliminary study in human volunteers.

Exogenous natural surfactant (ENS) labeled with (99m)Tc ((99m)Tc-ENS, 900-1110 MBq), a new radiopharmaceutical for ventilation scintigraphy, was nebulized during 3 min to five volunteers. For comparative purposes, (99m)Tc-diethylenetriamine pentaacetic acid (DTPA) was studied in the same way. (99m)Tc-ENS images were of at least the same quality as (99m)Tc-DTPA images. However, in smoking volunteers, the (99m)Tc-DTPA images show some areas that seemed to be not well-ventilated, although these areas appeared well-ventilated when the study was performed with (99m)Tc-ENS. These results suggest that (99m)Tc-ENS can be used for ventilation scintigraphy to allow the observation of some areas that cannot been visualized using (99m)Tc-DTPA as ventilation agent.

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.

[Zinc: concepts on an essential micronutrient]. / Cinc: conceptos actuales sobre un micronutriente esencial.

It is now well known that zinc is an essential micronutrient. Even though much information is available, there are many points that remain to be studied. zinc is absorbed by the intestine and transported in the plasma by albumin, forming a small exchangeable pool, which is rapidly exhausted even in cases of mild deficiency. zinc is essential for the activity of about 70 enzymes among other functions. Therefore, zinc deficiency produces a great number of clinical disorders, the symptoms of which may range between mild up to serious dysfunctions. These problems are corrected by a dietary supplementation with zinc. The lack of a reliable method to determine the zinc nutritional status reflects the problem to estimate the metal recommended dietary allowances, which are difficulty met specially by children, old persons and pregnant women which constitute the main risk groups. Although our knowledge concerning zinc toxicity is scarce, it is well known that the amounts of zinc that produces toxic effects are much higher than those that are contained in regular diets as well as in diets supplemented with this metal. Therefore, the need of the development of effective strategies, like food fortification with proper zinc compounds, appears as an attractive alternative in order to prevent and/or correct the deficiency of this vital element and to improve the health and the life quality of the whole population.

Cinc: conceptos actuales sobre un micronutriente esencial. / [Zinc: concepts on an essential micronutrient]

It is now well known that zinc is an essential micronutrient. Even though much information is available, there are many points that remain to be studied. zinc is absorbed by the intestine and transported in the plasma by albumin, forming a small exchangeable pool, which is rapidly exhausted even in cases of mild deficiency. zinc is essential for the activity of about 70 enzymes among other functions. Therefore, zinc deficiency produces a great number of clinical disorders, the symptoms of which may range between mild up to serious dysfunctions. These problems are corrected by a dietary supplementation with zinc. The lack of a reliable method to determine the zinc nutritional status reflects the problem to estimate the metal recommended dietary allowances, which are difficulty met specially by children, old persons and pregnant women which constitute the main risk groups. Although our knowledge concerning zinc toxicity is scarce, it is well known that the amounts of zinc that produces toxic effects are much higher than those that are contained in regular diets as well as in diets supplemented with this metal. Therefore, the need of the development of effective strategies, like food fortification with proper zinc compounds, appears as an attractive alternative in order to prevent and/or correct the deficiency of this vital element and to improve the health and the life quality of the whole population.

A spectrophotometric method for the determination of hydroperoxides in liposomes.

A modification of the Asakawa-Matsushita iodometric assay method for the determination of the content of lipid hydroperoxides was developed which permits the simultaneous processing of many samples of high lipid content. The method has the advantages of simplicity as well as good reproducibility, so it is not necessary to process standards with each determination. Our technique exceeds the sensitivity attained with other spectrophotometric determinations reported in the literature. The method requires the total elimination of water from the samples, and this was accomplished using an azeotropic mixture of ethanol:water of 96:4. The results obtained with liposomes indicate that the method is applicable to biological material limited to small volume samples, ranging 5-50 microliters. We want to emphasize that this method permits the study of the peroxidation process as function of time.

Oxidative damage, plasma antioxidant capacity, and glucemic control in elderly NIDDM patients.

A study of oxidative damage was made in elderly noninsulin-dependent diabetes mellitus (NIDDM) patients. A statistically significant increase in glucose and fructosamine was found in fasting NIDDM patients, as well as an increase in the oxidation induced by tert-butyl hydroperoxide. The Total Reactive Antioxidant Potential (TRAP) of the plasma was much reduced (p < .02) and the uricemia was unchanged. The erythrocytes of diabetic patients show greater basal oxidation products (p < .05), and the susceptibility of the diabetic erythrocytes to oxidation injury was also shown to increase in the oxidation induced by t-BOOH (p < .05). Linear regression studies showed that TRAP was associated directly with uric acid (p < .05) and inversely with fructosamine and with glucose (p < .03 and p < .05 respectively) in patients with NIDDM, but not in the controls. The levels of fructosamine were found to be related to the basal damage of the red blood cells (direct correlation, p < .001). This study suggest an useful approach to diabetic oxidative stress for clinical settings.

[The pulmonary surfactant system: physiology, pathologies associated with its alteration and exogenous administration as therapeutic and diagnostic agent]. / El sistema surfactante pulmonar: fisiología, patologías asociadas a su alteración y administración exógena como agente terapéutico y de diagnóstico.

Pulmonary surfactant is a lipoproteic mixture synthesized and secreted by alveolar type II cells. Its principal property is to reduce the surface tension by lining on the alveolar surface. Surfactant deficiency is the major factor responsible for the respiratory distress syndrome of the newborn (RDS) and the adult respiratory distress syndrome (ARDS). Since 1980, the exogenous administration of surfactant for the treatment of these syndromes is being studied. In this work the exogenous surfactant preparations, the delivery techniques and the dosing schedule is discussed. The utilization of the exogenous natural surfactant (ENS) as precursor of a radiopharmaceutical labeled with 99mTc (99mTc-ENS) for aerial lung scintigraphy is also discussed.

Oxidative stress and membrane fluidity in erythrocytes from patients with hemolytic uremic syndrome.

In order to investigate the implications of oxidative disturbances in the hemolysis associated with the Hemolytic Uremic Syndrome (HUS), basal levels of lipid peroxidation products, the response to t-butyl hydroperoxide induced damage and membrane fluidity were assayed by the technique of electron spin resonance in erythrocytes spin labeled with 5-Doxyl stearic acid obtained from eight children with HUS, during the 1st, 2nd, 4th and 12th weeks after diagnosis. During the acute phase of the disease, red blood cells (RBC) showed increased initial lipid peroxidation products, a higher susceptibility to oxidative insult and a lower membrane fluidity. All parameters reached control values the 12th week after diagnosis. The results suggest that in the acute phase of HUS, RBCs are exposed to an oxidative imbalance that could contribute to hemolysis directly through oxidative damage and/or by decreasing membrane fluidity.