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Biomedical subjects

F Sevilla

Publications and source records attributed to F Sevilla.

At least 19 recordsLinked to original sources

Roles for redox regulation in leaf senescence of pea plants grown on different sources of nitrogen nutrition.

Leaf senescence and associated changes in redox components were monitored in commercial pea (Pisum sativum L. cv. Phoenix) plants grown under different nitrogen regimes for 12 weeks until both nodules and leaves had fully senesced. One group of plants was inoculated with Rhizobium leguminosarum and grown with nutrient solution without nitrogen. A second group was not inoculated and these were grown on complete nutrient solution containing nitrogen. Leaf senescence was evident at 11 weeks in both sets of plants as determined by decreases in leaf chlorophyll and protein. However, a marked decrease in photosynthesis was observed in nodulated plants at 9 weeks. Losses in the leaf ascorbate pool preceded leaf senescence, but leaf glutathione decreased only during the senescence phase. Large decreases in dehydroascorbate reductase and catalase activities were observed after 9 weeks, but the activities of other antioxidant enzymes remained high even at 11 weeks. The extent of lipid peroxidation, the number of protein carbonyl groups and the level of H(2)O(2) in the leaves of both nitrate-fed and nodulated plants were highest at the later stages of senescence. At 12 weeks, the leaves of nodulated plants had more protein carbonyl groups and greater lipid peroxidation than the nitrate-fed controls. These results demonstrate that the leaves of nodulated plants undergo an earlier inhibition of photosynthesis and suffer enhanced oxidation during the senescence phase than those from nitrate-fed plants.

Aging↗

Location and effects of long-term NaCl stress on superoxide dismutase and ascorbate peroxidase isoenzymes of pea (Pisum sativum cv. Puget) chloroplasts.

The present work describes the intrachloroplast localization and the changes that took place in the thylakoid and stroma-located superoxide dismutases (SOD, EC 1.15.1.1) and ascorbate peroxidases (APX, EC 1.11.1.11), in response to long-term NaCl stress in Pisum sativum L. cv. Puget plants. Native PAGE using high chloroplast protein concentrations pointed to the presence of the two main Fe-SODs, together with CuZn-SODs, both in thylakoids and in the stroma. Western blot and immunogold labelling using the antibodies against chloroplastic Fe-SOD from Nuphar luteum also confirmed the chloroplastic localization of a Fe-SOD. Thylakoidal Fe-SOD activity was induced by a NaCl concentration as low as 70 mM, while CuZn-SOD was induced at 90 mM, although in severe stress conditions (110 mM) both activities were similar to the levels at 90 mM NaCl. NaCl stress also induced stromatic Fe-SOD and CuZn-SOD activities, although these inductions only started at higher NaCl concentration (90 mM) and were significant at 110 mM NaCl. The increase in activity of both Fe-SODs was matched by an increase in Fe-SOD protein. Chloroplastic APX isoenzymes behaved differently in thylakoids and stroma in response to NaCl. A significant increase of stromal APX occurred at 70 mM, whereas the thylakoidal APX activity was significantly and progressively lost in response to NaCl stress (70-110 mM). A significant increase in the H2O2 content of chloroplasts during stress and a reduction in the ascorbate level at 90 mM NaCl also took place, although the oxidized ascorbate pool at the highest NaCl concentration did not show significant changes. These results suggest that the loss of thylakoidal APX may be an important factor in the increase in chloroplastic H2O2, which also results from the increased thylakoid and stroma-located Fe-SOD and CuZn-SOD activities. This H2O2 may be involved in the induction of stromal APX. The up-regulation of the above enzymes in the described stress conditions would contribute to the adaptation of cv. Puget plants to moderate NaCl stress.

Adaptation, Physiological↗

Antioxidant systems and O(2)(.-)/H(2)O(2) production in the apoplast of pea leaves. Its relation with salt-induced necrotic lesions in minor veins.

The present work describes, for the first time, the changes that take place in the leaf apoplastic antioxidant defenses in response to NaCl stress in two pea (Pisum sativum) cultivars (cv Lincoln and cv Puget) showing different degrees of sensitivity to high NaCl concentrations. The results showed that only superoxide dismutase, and probably dehydroascorbate reductase (DHAR), were present in the leaf apoplastic space, whereas ascorbate (ASC) peroxidase, monodehydroascorbate reductase (MDHAR), and glutathione (GSH) reductase (GR) seemed to be absent. Both ASC and GSH were detected in the leaf apoplastic space and although their absolute levels did not change in response to salt stress, the ASC/dehydroascorbate and GSH to GSH oxidized form ratios decreased progressively with the severity of the stress. Apoplastic superoxide dismutase activity was induced in NaCl-treated pea cv Puget but decreased in NaCl-treated pea cv Lincoln. An increase in DHAR and GR and a decrease in ASC peroxidase, MDHAR, ASC, and GSH levels was observed in the symplast from NaCl-treated pea cv Lincoln, whereas in pea cv Puget an increase in DHAR, GR, and MDHAR occurred. The results suggest a strong interaction between both cell compartments in the control of the apoplastic ASC content in pea leaves. However, this anti-oxidative response does not seem to be sufficient to remove the harmful effects of high salinity. This finding is more evident in pea cv Lincoln, which is characterized by a greater inhibition of the growth response and by a higher rise in the apoplastic hydrogen peroxide content, O(2)(.-) production and thiobarbituric acid-reactive substances, and CO protein levels. This NaCl-induced oxidative stress in the apoplasts might be related to the appearance of highly localized O(2)(.-)/H(2)O(2)-induced necrotic lesions in the minor veins in NaCl-treated pea plants. It is possible that both the different anti-oxidative capacity and the NaCl-induced response in the apoplast and in the symplast from pea cv Puget in comparison with pea cv Lincoln contributes to a better protection of pea cv Puget against salt stress.

Adaptation, Physiological↗

Differential response of antioxidative enzymes of chloroplasts and mitochondria to long-term NaCl stress of pea plants.

In this work the activity of superoxide dismutase (SOD) and the enzymes of the ascorbate-glutathione (ASC-GSH) cycle were investigated in chloroplasts and mitochondria from leaves of Pisum sativum L. cv. Puget after 15 days treatment with 0-130 mM NaCl. The main chloroplastic SOD activity was due to CuZn-SOD II, which was increased significantly (about 1.7-fold) by NaCl, although during severe NaCl stress (110-130 mM) chloroplastic Fe-SOD exhibited a stronger enhancement in its activity (about 3.5-fold). A sudden induction in chloroplastic APX, DHAR and GR was also caused by NaCl (70-110 mM), but not by the highest salt concentration (130 mM), at which GR and DHAR activities were similar to the control values and APX decreased. In addition, the H2O2 concentration and lipid peroxidation of membranes increased significantly, 3.5- and 7-fold, respectively, in chloroplasts under severe NaCl stress. In purified mitochondria DHAR and GR were significantly induced only at 90 and 130 mM NaCl, respectively, although DHAR activity was below control values in the highest NaCl concentrations. APX and MDHAR activities started their response to salt in mild NaCl conditions (70 mM) and increased significantly with the severity of the stress. Mn-SOD was induced only under severe NaCl concentrations. The mitochondrial H2O2 and lipid peroxidation were increased at the highest NaCl concentration although to a lesser extent (about 2-2.5-fold) than in chloroplasts, whereas the increase in carbonyl protein contents was higher in mitochondria. The results suggest that the degree of enhanced tolerance to NaCl seems to require the induction of specific isoforms, depending on the different organelles.

Antioxidants↗

Adenomyoepithelioma of the breast: report of two cases with prominent cystic changes and intranuclear inclusions.

Adenomyoepithelioma is a rare breast tumor. Histologically it may disclose different patterns of growth, and some additional features may result in diagnostic errors. We describe 2 cases of adenomyoepithelioma of the breast initially examined by fine-needle aspiration biopsy (FNAB). Cytologic features included hypercellularity, clusters of epithelial and myoepithelial cells with occasional intranuclear inclusions, prominent apocrine metaplasia, and foam cells. Histologically, both tumors were diagnosed as adenomyepithelioma tubular-variant, with prominent myoepithelial clear cells, apocrine metaplasia, and foci of squamous metaplasia. Immunohistochemically, the tumors showed strong positivity for keratins CAM 5.2, AE1/AE3, and EMA in the epithelial component, while the myoepithelial cells reacted with muscle-specific actin (A14 and HHF35) and S-100 protein. We point out that FNAB in this rare tumor may exhibit a varied spectrum of cells that may result in confusion with other lesions, and we call attention to the presence of intranuclear inclusions. The latter observation in the present cases by cytologic and histologic assessment provides and additional feature to the morphological characteristics of adenomyoepithelioma of the breast.

Actins↗

Intramammary lymph node involvement by mycosis fungoides diagnosed by fine-needle aspiration biopsy.

We report on a case of a patient with diagnosis of mycosis fungoides (MF) who subsequently developed enlargement of the intramammary lymph nodes, which were demonstrated to be involved by tumor on fine-needle aspiration (FNA) cytology. The patient was a 66-yr-old female who had recently noticed the appearance of circumscribed bilateral breast nodules. A mammographic study showed clearly outlined nodules of low density. FNA showed atypical small and large lymphocytes with cerebriform appearance, consistent with intramammary lymph node infiltration by MF. To our knowledge this is the first report of intramammary palpable lymph nodes involved by MF and confirmed by FNA. This case provides support for the use of fine-needle aspiration biopsy as an accurate method for staging MF successfully.

Aged↗

Role of the ascorbate-glutathione cycle of mitochondria and peroxisomes in the senescence of pea leaves

We investigated the relationship between H2O2 metabolism and the senescence process using soluble fractions, mitochondria, and peroxisomes from senescent pea (Pisum sativum L.) leaves. After 11 d of senescence the activities of Mn-superoxide dismutase, dehydroascorbate reductase (DHAR), and glutathione reductase (GR) present in the matrix, and ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR) activities localized in the mitochondrial membrane, were all substantially decreased in mitochondria. The mitochondrial ascorbate and dehydroascorbate pools were reduced, whereas the oxidized glutathione levels were maintained. In senescent leaves the H2O2 content in isolated mitochondria and the NADH- and succinate-dependent production of superoxide (O2.-) radicals by submitochondrial particles increased significantly. However, in peroxisomes from senescent leaves both membrane-bound APX and MDHAR activities were reduced. In the matrix the DHAR activity was enhanced and the GR activity remained unchanged. As a result of senescence, the reduced and the oxidized glutathione pools were considerably increased in peroxisomes. A large increase in the glutathione pool and DHAR activity were also found in soluble fractions of senescent pea leaves, together with a decrease in GR, APX, and MDHAR activities. The differential response to senescence of the mitochondrial and peroxisomal ascorbate-glutathione cycle suggests that mitochondria could be affected by oxidative damage earlier than peroxisomes, which may participate in the cellular oxidative mechanism of leaf senescence longer than mitochondria.

Journal Article↗

The Lactational Amenorrhea Method (LAM): a postpartum introductory family planning method with policy and program implications.

It is well accepted that breastfeeding contributes significantly to child survival and child nutrition. Healthful child spacing is associated with improved birth outcomes and maternal recovery. On a population basis, breastfeeding may contribute more to birth spacing than all family planning use combined in many countries. However, while breastfeeding does provide a period of infertility, until recently, there was no reliable way for an individual woman to capitalize on this lactational infertility for her own efficacious child spacing. The Lactational Amenorrhea Method (LAM) is a new introductory family planning method that simultaneously promotes child spacing and breastfeeding, with its optimal nutrition and disease preventive benefits for the infant. LAM, as it is called, is based on the utilization of lactational infertility for protection from pregnancy and indicates the time for the introduction of a complementary family planning method. LAM is recommended for up to six months postpartum for women who are fully or nearly fully breastfeeding and amenorrheic, and relies on the maintenance of appropriate breastfeeding practices to prolong lactational infertility, with the concomitant delay in menses return. A recent clinical trial confirmed the theoretical 98% or higher effectiveness of the method and field trials are demonstrating its acceptability. Nonetheless, some demographers and family planning organizations continue to debate its value. The development, efficacy, and sequelae of the method are presented using data from several studies by the authors.

Amenorrhea↗

Purification of an iron-containing superoxide dismutase from a citrus plant, Citrus limonum R.

A cyanide-insensitive superoxide dismutase was purified to apparent homogeneity from lemon leaves (Citrus limonum R). The enzyme was isolated from leaf extracts by ammonium sulfate salting-out, and ion-exchange, gel filtration and hydroxylapatite column chromatography. The purified Fe-SOD had a specific activity of about 1,500 U/mg and represents approximately 1.6% of the total soluble protein in lemon leaf extracts. A molecular weight of 47,500 was determined for the enzyme. Analytical gel electro-focusing of the purified preparation revealed the presence of two isozymes with pI values of 5.13 and 4.98. Metal analysis showed the presence of 1 g-atom of iron and 0.5 g-atom of manganese per mol of enzyme. The visible and UV absorption spectra of the Citrus enzyme were similar to those reported for other iron-containing SODs from different origins. The significance of the presence of Fe-SOD in higher plants is briefly discussed.

Chromatography, Ion Exchange↗

Nutritional effect and expression of SODs: induction and gene expression; diagnostics; prospective protection against oxygen toxicity.

The effect of micronutrient stress (either deficiency or toxicity) on the expression of different superoxide dismutase isoenzymes in plants is reviewed. The induction of Fe-SOD and Mn-SOD by different metals and the potential use of the metalloenzyme system SOD for the appraisal of the micronutrient status of plants, is examined. At subcellular level, evidence for the participation of peroxisomal SOD in the molecular mechanism of plant tolerance to Cu is presented, and the activated oxygen-dependent toxicity of a xenobiotic (clofibrate) in plant peroxisomes is examined.

Enzyme Induction↗

Somatostatin analog SMS 201-995 and insulin needs in insulin-dependent diabetic patients studied by means of an artificial pancreas.

SMS 201-995 is a new somatostatin analog which is 10-60 times more potent and specific than somatostatin as an inhibitor of GH and insulin release. The aim of this study was to assess its value as an adjunct to insulin therapy in insulin-dependent diabetic- (IDD) patients. Six IDD patients were studied. Their average insulin doses ranged from 22-46 U/day, and hemoglobin A1c levels varied between 6.5-11.5%. Two patients had background retinopathy and mild sensorimotor neuropathy. After 12 h of glucemic stabilization, the patients were kept normoglycemic by connecting them to the Biostator-GCIIS. The study entailed two parts in random order, in which standardised mixed meals were administered at 0800, 1400, and 2000 h with or without sc bolus injections of 50 micrograms SMS 201-995 immediately before meal ingestion. Plasma free insulin, C-peptide, GH, and glucagon were measured by RIA. Postprandial hyperglycemia was significantly diminished by SMS 201-995 after breakfast, lunch, and dinner. Insulin requirements, both total and 2-h postprandially, decreased significantly with a parallel reduction in free insulin levels. Postprandial glucagon levels also significantly decreased, but GH profiles were similar. In conclusion, the somatostatin analog SMS 201-995 has a potential value as an adjunct to insulin in the management of IDD patients.

Adult↗

Pilot study to detect neurologic disease in Ecuador among a population with a high prevalence of endemic goiter.

A census, uniform screening questionnaire, and simple screening neurologic examination were administered in a door-to-door survey to residents of Quiroga, Ecuador, a rural community in the Andes Mountains. The screening procedures had been pretested to assure a high level of sensitivity for detecting children and adults with major neurologic disease. A total of 1,113 participated in the study. Of these, 399 had responses or findings suggesting the presence of neurologic disease. These individuals were then examined by a neurologist, who used fixed diagnostic criteria. The prevalence ratios (per 1,000) for the most common neurologic conditions identified in this survey are: recurrent/persistent severe headache = 68.3, and epilepsy = 17.1.

Adolescent↗