Publications by authors named "Raquel Ruiz"

18 Publications

  • Page 1 of 1

The effect of omitting axillary dissection and the impact of radiotherapy on patients with breast cancer sentinel node macrometastases: a cohort study following the ACOSOG Z0011 and AMAROS trials.

Breast Cancer Res Treat 2021 Aug 4;189(1):111-120. Epub 2021 Jun 4.

Bellvitge University Hospital: Hospital Universitari de Bellvitge, Hospitalet de Llobregat, Spain.

Purpose: To report the outcomes of implementing the ACOSOG Z0011 and AMAROS trials relevant to clinical practice, and to define target groups in whom to avoid or recommend axillary radiotherapy (ART). We also aimed to analyse the reduction in morbidity when axillary lymph node dissection (ALND) was omitted.

Methods: A retrospective cohort study of T1-T2 patients with macrometastases at sentinel lymph node (SLN) who were treated between 2011 and 2020. Breast surgery included either lumpectomy or mastectomy. Patients with ≤ 2 positive SLN were divided into two cohorts by whether they received ART or not. Survival outcomes and morbidity were analysed by Kaplan-Meyer curves and Cox-regression, respectively.

Results: 260 pN1a patients were included and ALND was avoided in 167 (64.2%). According the Z0011 results, 72 (43.1%) received no further ART; and based on AMAROS criteria 95 (56.9%) received ART. Median follow-up was 54 months. The 5-year overall survival was 96.8% in the non-RT cohort and 93.4% in the RT cohort (p = 0.19), while the respective 5-year disease-free survivals were 100% and 92.3% (p = 1.06). Lymphedema developed in 3.6% of patients after SLNB versus 43% after ALND (OR 20.25; 95%CI 8.13-50.43). Decreased upper-extremity range of motion appeared in 8.4% of patients after SLNB versus 31.2% after ALND (OR 4.95; 95%CI 2.45-9.98%).

Conclusions: Our study confirms that omitting ALND is safe and has high survival rates in patients with T1-T2 tumours and ≤ 2 positive SLNs. Adding ART could be a treatment option for patients who present other risk factors. Avoiding ALND with or without ART was associated with significantly less arm morbidity.
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http://dx.doi.org/10.1007/s10549-021-06274-9DOI Listing
August 2021

A Pea ( L.) Seed Vicilins Hydrolysate Exhibits PPARγ Ligand Activity and Modulates Adipocyte Differentiation in a 3T3-L1 Cell Culture Model.

Foods 2020 Jun 16;9(6). Epub 2020 Jun 16.

Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín (EEZ, CSIC), 18008 Granada, Spain.

Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea ( L.) seed meal proteins (albumins, legumins and vicilins) were isolated, submitted to a simulated gastrointestinal digestion, and the effects of their hydrolysates (pea albumins hydrolysates (PAH), pea legumins hydrolysates (PLH) and pea vicilin hydrolysates (PVH), respectively) on 3T3-L1 murine pre-adipocytes were investigated. The pea vicilin hydrolysate (PVH), but not native pea vicilins, increased lipid accumulation during adipocyte differentiation. PVH also increased the mRNA expression levels of the adipocyte fatty acid-binding protein (aP2) and decreased that of pre-adipocyte factor-1 (Pref-1) (a pre-adipocyte marker gene), suggesting that PVH promotes adipocyte differentiation. Moreover, PVH induced adiponectin and insulin-responsive glucose transporter 4 (GLUT4) and stimulated glucose uptake. The expression levels of peroxisome proliferator-activated receptor γ (PPARγ), a key regulator of adipocyte differentiation, were up-regulated in 3T3-L1 cells treated with PVH during adipocyte differentiation. Finally, PVH exhibited PPARγ ligand activity. Lactalbumin or other pea hydrolysates (PAH, PLH) did not exhibit such effects. These findings show that PVH stimulates adipocyte differentiation via, at least in part, the up-regulation of PPARγ expression levels and ligand activity. These effects of PVH might be relevant in the context of the beneficial health effects of legume consumption in obesity-associated metabolic disorders.
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http://dx.doi.org/10.3390/foods9060793DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353609PMC
June 2020

Ultrasound examination at term for predicting the outcome of delivery in women with a previous cesarean section.

J Matern Fetal Neonatal Med 2016 Dec 3;29(23):3870-4. Epub 2016 Mar 3.

a Department of Obstetrics and Gynecology , Corporació Sanitària Parc Taulí , Universitat Autònoma de Barcelona , Sabadell , Barcelona , Spain.

Objective: To evaluate if ultrasound variables at term are associated with the mode of delivery in women with previous cesarean section (PCS).

Methods: This was a prospective study of singleton pregnant women who planned a trial of vaginal birth after cesarean delivery. Cervical length, posterior cervical angle, head-perineum distance, and estimated fetal weight were measured at 37-39 weeks of gestation.

Results: One hundred forty-four pregnancies were examined and vaginal delivery was achieved in 98 women (73%). Logistic regression analysis identified cervical length, head-perineum distance, age, previous vaginal delivery, previous cesarean for dystocia, and Bishop score as predictors of vaginal delivery. Combining ultrasound and clinical parameters, two models for risk scoring that differ in the variable Bishop score or cervical length were constructed. The AUC of these models was 0.867 and 0.855, respectively.

Conclusions: In women with a PCS, measurement of cervical length and head-perineum distance at term is associated with the mode of delivery. A combination of clinical and sonographic parameters at term can predict the likelihood of vaginal delivery.
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http://dx.doi.org/10.3109/14767058.2016.1149566DOI Listing
December 2016

Pea (Pisum sativum L.) seed albumin extracts show anti-inflammatory effect in the DSS model of mouse colitis.

Mol Nutr Food Res 2015 Apr 2;59(4):807-19. Epub 2015 Mar 2.

CIBER-EHD, Department of Pharmacology, ibs. GRANADA, Center for Biomedical Research (CIBM), University of Granada, Granada, Spain.

Scope: This study investigates the preventive effects of two pea (Pisum sativum) seed albumin extracts, either in the presence (pea seed extract [PSE]) or absence (albumin fraction from PSE [AF-PSE]) of soluble polysaccharides, in the dextran sodium sulfate (DSS) induced colitis in mice.

Methods And Results: Male C57BL/6J mice were assigned to five groups: one noncolitic and four colitic. Colitis was induced by incorporating DSS (3.5%) in the drinking water for 4 days, after which DSS was removed. Treated groups received orally PSE (15 g/kg⋅day), or AF-PSE (1.5 g/kg⋅day), or pure soy Bowman-Birk inhibitor (BBI; 50 mg/kg⋅day), starting 2 wk before colitis induction, and maintained for 9 days after. All treated groups showed intestinal anti-inflammatory effect, evidenced by reduced microscopic histological damage in comparison with untreated colitic mice. The treatments ameliorated the colonic mRNA expression of different proinflammatory markers: cytokines, inducible enzymes, metalloproteinases, adhesion molecules, and toll-like receptors, as well as proteins involved in maintaining the epithelial barrier function. Furthermore, the administration of PSE, AF-PSE, or soy BBI restored bacterial counts, partially or totally, to values in healthy mice.

Conclusion: PSE and AF-PSE ameliorated DSS-induced damage to mice, their effects being due, at least partially, to the presence of active BBI.
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http://dx.doi.org/10.1002/mnfr.201400630DOI Listing
April 2015

Massive left atrium thrombus.

Int J Cardiovasc Imaging 2014 Jan 10;30(1):67. Epub 2013 Oct 10.

Department of Cardiology, Hospital Universitario Fundación Alcorcón, Av Budapest, 1., 28922, Alcorcón, Madrid, Spain,

The image shows a left atrium completely occupied by a giant non-homogeneous, irregularly-surfaced mass, obstructing the disc motion, mimicking severe mitral prosthetic stenosis.
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http://dx.doi.org/10.1007/s10554-013-0299-2DOI Listing
January 2014

Characterization of pea (Pisum sativum) seed protein fractions.

J Sci Food Agric 2014 Jan 8;94(2):280-7. Epub 2013 Jul 8.

Physiology and Biochemistry of Animal Nutrition (EEZ, CSIC), Profesor Albareda 1, E-18008, Granada, Spain.

Background: Legume seed proteins have to be chemically characterized in order to properly link their nutritional effects with their chemical structure.

Results: Vicilin and albumin fractions devoid of cross-contamination, as assessed by mass peptide fingerprinting analysis, were obtained from defatted pea (Pisum sativum cv. Bilbo) meal. The extracted protein fractions contained 56.7-67.7 g non-starch polysaccharides kg⁻¹. The vicilin fraction was higher than legumins in arginine, isoleucine, leucine, phenylalanine and lysine. The most abundant amino acids in the albumin fraction were aspartic acid, glutamic acid, lysine and arginine, and the amounts of methionine were more than double than those in legumins and vicilins. The pea albumin fraction showed a clear enrichment of protease inhibitory activity when compared with the seed meal. In vitro digestibility values for pea proteins were 0.63 ±  0.04, 0.88 ±  0.04 and 0.41 ±  0.23 for legumins, vicilins and albumins respectively.

Conclusion: Vicilin and albumin fractions devoid of cross-contamination with other proteins were obtained from pea seed meal. The vicilin fraction also contained low amounts of soluble non-starch polysaccharides and was enriched in isoleucine, leucine, phenylalanine and lysine. In vitro digestibility values for pea proteins were similar or even numerically higher than those for control proteins.
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http://dx.doi.org/10.1002/jsfa.6250DOI Listing
January 2014

Garlic derivatives (PTS and PTS-O) differently affect the ecology of swine faecal microbiota in vitro.

Vet Microbiol 2010 Jul 28;144(1-2):110-7. Epub 2009 Dec 28.

Dpto de Fisiología y Bioquímica de la Nutrición Animal IFNA, EEZ, CSIC, Profesor Albareda 1, 18008 Granada, Spain.

A number of in vitro experiments were designed to evaluate the effects of two different industrial products, namely PROALLIUM-S-DMC and PROALLIUM-SO-DMC (DMC Research Center, Granada, Spain), obtained from garlic (Allium sativum) on the faecal microbiota of pigs. The effects of three different concentrations (50, 200 and 400 ppm) of the active compounds (PTS and PTS-O, respectively) from both industrial products on the gastrointestinal microbiota of pigs were tested. Growth medium without any additive (0 ppm) was used as control. Predominant bacterial groups (total aerobes, total anaerobes, lactobacilli, bifidobacteria, coliforms, enterobacteria, bacteroides and clostridia) were studied. Results showed that both PTS and PTS-O have significant (P<0.01) antimicrobial activity against every group studied, although enterobacteria and coliforms were the most affected populations (P<0.01). Time kill curves for Escherichia coli and Salmonella typhimurium, two common pathogens of pigs, showed that both compounds had a bactericidal effect against these strains. For the bacterial groups here studied, the antimicrobial effect of PTS-O was significantly (P<0.001) stronger than that of PTS. Trials in vivo are in course to study the potential use of these products as alternatives to antibiotics in pig feeds.
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http://dx.doi.org/10.1016/j.vetmic.2009.12.025DOI Listing
July 2010

Anti-carcinogenic soyabean Bowman-Birk inhibitors survive faecal fermentation in their active form and do not affect the microbiota composition in vitro.

Br J Nutr 2009 Apr;101(7):967-71

Department of Biochemistry and Physiology of Animal Nutrition, Institute of Animal Nutrition, Estación Experimental del Zaidín (CSIC), Profesor Albareda 1, 18008 Granada, Spain.

Bowman-Birk inhibitor (BBI) from soyabeans is a naturally occurring protease inhibitor with potential anti-inflammatory and chemopreventive properties within the gastrointestinal tract (GIT). In a previous paper, we reported that significant amounts of BBI-related proteins reach the terminal ileum functionally and biologically active. We have now investigated: (a) if soyabean BBI is biotransformed by faecal microbiota which would reduce its potential colorectal chemopreventive properties and (b) the potential influence of this protease inhibitor on the modulation of faecal microbiota. In vitro incubation studies of native soyabean BBI at a physiological level (93 microM) with mixed faecal samples of pigs for 24 h at 37 degrees C demonstrated that BBI remains active and its intrinsic trypsin and chymotrypsin inhibitory activities were not significantly influenced by the enzymic or metabolic activity of faecal microbiota. Soyabean BBI did not affect the growth of the different bacterial groups studied (lactobacilli, bifidobacteria, bacteroides, coliforms, enterobacteria, clostridia and total anaerobes). It was concluded that protease inhibitory activities, intrinsically linked to the chemopreventive properties of soyabean BBI, were largely unaffected by faecal microbiota in vitro. BBI retains significance, therefore, as a bioactive compound in the human GIT.
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http://dx.doi.org/10.1017/s0007114508057590DOI Listing
April 2009

Effects of cellulite treatment with RF, IR light, mechanical massage and suction treating one buttock with the contralateral as a control.

J Cosmet Laser Ther 2008 Dec;10(4):193-201

Instituto Medico Vilafortuny/Antoni De Gimbernat Foundation, Cambrils, Spain.

Background And Objectives: A system that combines bipolar radio frequency (RF) and intense infrared light (IR) together with mechanical massage and suction has recently been reported as being efficient for cellulite treatment. The present split study was designed to evaluate the efficacy of such a system through various treatments of cellulite located on the buttocks.

Methods: Ten patients were enrolled for 12 sessions of 30 minutes each performed over one buttock, the other buttock serving as an untreated control. Sessions were conducted twice a week for a period of 12 weeks. Clinical photography and profilometry were carried out to assess textural changes before (baseline) and 2 months after the final treatment. Histopathology was performed at baseline, 2 hours after the first session, and just before the 12th session and 2 months thereafter.

Results: All patients noted improvement in the treated buttock before the final session, which was maintained at the 2-month assessment. Improved skin appearance was noticed after the first session and was maintained throughout the study. All patients were satisfied with the results and requested further treatment in order to balance the results in both buttocks. Random histological analyses suggested dermal firmness, fibre compaction and tightening of skin layers, including the subcutis, as possible reasons for the effects achieved. The authors recognize that the small number of participants limits the statistical power of the study.

Conclusions: Treatment sessions with the combined RF, IR light and mechanical massage and suction system were complication free, produced improvements in the overall cellulite appearance and skin condition, suggesting that further treatment sessions for maintenance could sustain patient satisfaction index (SI) and lead to lasting results. Based on the good results in the limited trial population, further studies with larger patient populations are warranted.
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http://dx.doi.org/10.1080/14764170802524403DOI Listing
December 2008

[A new fish species Hemibrycon (Characiformes: Characidae)].

Rev Biol Trop 2006 Mar;54(1):209-17

Universidad del Quindío, Laboratorio de Ictiología, A.A. 460, Armenia, Quindío, Colombia.

Hemibrycon pautensis (Characiformes, Characidae), a new fish species from Paute River, eastern Ecuador is described. Diagnostic characteristics: eight to nine branched rays in the dorsal fin (vs. six to seven), and 27 - 28 in the anal fin (vs. 16 - 26, except in H. dariensis which presents 22 - 27, in H. metae 26 - 31 and H. jabonero 23 - 28); a no occurrence of dorsal pharyngeal plate (vs. occurrence); a cartilaginous and divided-in-two basihial (vs. an osseous base and a cartilaginous upper part). Hemibrycon pautensis resembles H. metae by its oblique external edge of the pelvic fins. They can be distinguished by the position of the pectoral fins in relation to the snout (38.24-41.6% in H. pautensis vs. 21.21-25.87) and by the position of the pectoral fins in relation to the origin of the dorsal fin (20.95-24.30 in Hemibrycon pautensis vs. 35.89-42.63), and by the number of proximate radials in the pectoral girdle (five in Hemibrycon pautensis vs. three to four). In addition, the geographic distribution of H. metae is restricted to the upper part of the Meta River in Colombia and can be distinguished of H. boquiae by: the number of scales between the lateral-line and the origin of the dorsal fin (eight in H. pautensis vs. 5-7); the distance between the snout and the pelvic fins (38.00-42.90 % in H. pautensis vs. 42.9-46.19%); the pelvic fins length (13.77-17.96% in H. pautensis vs. 10.72-13.21%); and the snout length (21.34-27.88 in H. pautensis vs. 26.92-33.66%).
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March 2006

Complexity in efflux pump control: cross-regulation by the paralogues TtgV and TtgT.

Mol Microbiol 2007 Dec 5;66(6):1416-28. Epub 2007 Nov 5.

Department of Environmental Protection, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Granada, Spain.

Pseudomonas putida DOT-T1E, known for its high tolerance to solvents, possesses three Resistance-Nodulation-Cell Division-type (RND) efflux pumps, namely TtgABC, TtgDEF and TtgGHI, which are involved in the active extrusion of solvents. Expression of the ttgABC and ttgGHI operons was previously shown to be regulated by the adjacently encoded repressors, TtgR and TtgV, respectively. Upstream of the third RND operon, ttgDEF, is located a putative regulator gene, ttgT. In this study, TtgT is shown to bind to the promoter region of the ttgDEF operon, and to be released from DNA in the presence of organic solvents. In vitro studies revealed that TtgV and TtgT bind the same operator sites in both the ttgDEF and the ttgGHI promoters. However, the affinity of TtgV for the ttgDEF operator was higher than that of TtgT, which, together with the fact that the ttgV promoter seems to be almost twice stronger than the ttgT promoter, explains why TtgV takes over in the regulation of the two efflux pump operons. The functional replacement of the cognate, chromosomally encoded TtgT by the plasmid-encoded paralogue TtgV illustrates a new mode of efflux pump regulation of which the physiological relevance is discussed.
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http://dx.doi.org/10.1111/j.1365-2958.2007.06004.xDOI Listing
December 2007

[A new Ecuadorian species of the Andean fish genus Grundulus (Characiformes: Characidae)].

Rev Biol Trop 2005 Sep-Dec;53(3-4):537-44

Universidad del Quindio, laboratorio de ictiologia, A.A. 460, Armenia, Quindio, Colombia.

The new species Grundulus quitoensis is described from El Voladero Lake, El Angel Biological Reserve, Northern Ecuador. The new taxon can be distinguished from all congeners by the following characters, respectively: a low number of unbranched rays in the anal fin (three vs. four-five) and in pectoral fins (on vs. two - three), a higher number of dentary teeth (12-14 vs. 8-10), a second infraorbital about three times larger than the first (vs. about two times larger), a third infraorbital which is not in contact with the preopercle (vs. in contact with preopercle), the presence of four small fenestrae in the infero-lateral bone dentary (vs. one - two); and a premaxilla which presents a long lateral process (vs. short), a maxilla with two notches in the infero-lateral surface (vs. one notch in the infero-lateral surface).
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July 2007

Systemic pathological alterations caused by Philodryas patagoniensis colubrid snake venom in rats.

Toxicon 2006 Oct 30;48(5):520-8. Epub 2006 Jun 30.

Cátedra de Química Biológica I, Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste (UNNE), Av. Libertad 5470, Corrientes 3400, Argentina.

Very little is known about the systemic effects caused by Philodryas patagoniensis colubrid snake venom. In this work, this venom was tested for its ability to induce histopathological changes in rats after its intramuscular, subcutaneous or intravenous administration, by light microscopic examination of some organs (cerebellum, cerebrum, lung, liver, kidney and heart). Four rats were used for each dose of 0.23, 0.45 and 0.90 mg of venom in 0.3 ml of phosphate-buffered saline solution (pH 7.4). Aliquots of blood were withdrawn at different time intervals for enzymatic determination of alanine aminotransferase, aspartate aminotransferase and creatine kinase levels. After 2h the animals were killed by an overdose of anesthetic, and samples of kidney, heart, liver, lung, cerebrum and cerebellum were taken to microscopic examination (hematoxylin and eosin stain). Histologically, no abnormality was observed in heart tissue, in none of the administration routes of the venom used. However, histological observations showed multifocal hemorrhage in cerebellum, cerebrum and lung sections, severe peritubular capillary congestion in kidney sections and hydropic degeneration in liver sections, when venom was administrated intravenously. The subcutaneous route showed similar results to the previous one, with the exception of cerebellar hemorrhage. Intramuscularly, neither cerebral nor cerebellar hemorrhage was observed. Plasma alanine aminotransferase and aspartate aminotransferase increased levels were demonstrated, mainly when venom was administered intravenously or subcutaneously. Our results suggest that P. patagoniensis venom induces moderate histopathological changes in vital organs of rats. These changes are initiated at early stages of the envenomation and may be associated with a behavioral or functional abnormality of those organs during envenoming.
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http://dx.doi.org/10.1016/j.toxicon.2006.06.013DOI Listing
October 2006

The multidrug efflux regulator TtgV recognizes a wide range of structurally different effectors in solution and complexed with target DNA: evidence from isothermal titration calorimetry.

J Biol Chem 2005 May 13;280(21):20887-93. Epub 2005 Mar 13.

Department of Biochemistry and Molecular and Cellular Biology of Plants, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Profesor Albareda, 1, E-18008 Granada, Spain.

TtgV modulates the expression of the ttgGHI operon, which encodes an efflux pump that extrudes a wide variety of chemicals including mono- and binuclear aromatic hydrocarbons, aliphatic alcohols, and antibiotics of dissimilar chemical structure. Using a 'lacZ fusion to the ttgG promoter, we show that the most efficient in vivo inducers were 1-naphthol, 2,3-dihydroxynaphthalene, 4-nitrotoluene, benzonitrile, and indole. The thermodynamic parameters for the binding of different effector molecules to purified TtgV were determined by isothermal titration calorimetry. For the majority of effectors, the interaction was enthalpy-driven and counterbalance by unfavorable entropy changes. The TtgV-effector dissociation constants were found to vary between 2 and 890 mum. There was a relationship between TtgV affinity for the different effectors and their potential to induce gene expression in vivo, indicating that the effector binding constant is a major determinant for efficient efflux pump gene expression. Equilibrium dialysis and isothermal titration calorimetry studies indicated that a TtgV dimer binds one effector molecule. No evidence for the simultaneous binding of multiple effectors to TtgV was obtained. The binding of TtgV to a 63-bp DNA fragment containing its cognate operator was tight and entropy-driven (K(D) = 2.4 +/- 0.35 nm, DeltaH = 5.5 +/- 0.04 kcal/mol). The TtgV-DNA complex was shown to bind 1-napthol with an affinity comparable with the free soluble TtgV protein, K(D) = 4.8 +/- 0.19 and 3.0 +/- 0.15 mum, respectively. The biological relevance of this finding is discussed.
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http://dx.doi.org/10.1074/jbc.M500783200DOI Listing
May 2005

Mice plasma fibrinogen consumption by thrombin-like enzyme present in rattlesnake venom from the north-east region of Argentina.

Medicina (B Aires) 2004 ;64(6):509-17

Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste, Corrientes, Argentina.

Due to variability of venom components from the same species of snakes that inhabit different regions, particular properties of the venom of Crotalus durissus terrificus that inhabits the North-East of Argentina were studied. Gyroxin, a thrombin-like enzyme, was isolated from this venom by gel filtration and affinity chromatography, it was found to be homogeneous according to SDS-PAGE, with a molecular weight of 33 kDa. "Gyroxin syndrome" in mice was tested and it showed changes in the animal behavior, confirming that the isolated thrombin-like enzyme is gyroxin. Effects of this enzyme and the crude venom on mice plasmatic fibrinogen levels were determined. The mice plasma fibrinogen decreased rapidly until incoagulability during the first hour after thrombin-like enzyme injection, then reaching its normal level 10 hours after injection; whereas crude venom resulted in a 60% decrease of the mice plasma fibrinogen, reaching its normal level after the same period of time. After 1 hour of gyroxin inoculation, intravascular coagulation was observed in histological cuttings of lung, cardiac muscle and liver. The isolated enzyme showed strong hydrolyzing activity on fibrinogen and fibrin in vitro, whereas the crude venom exhibited weak hydrolyzing activity on both substrates. It is probable that this very low activity is due to the low percentage of the enzyme in the crude venom. Decreasing of plasmatic fibrinogen levels may be due to either the coagulant or hydrolyzing actions of the enzyme.
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October 2005

Edematogenic and myotoxic activities of the Duvernoy's gland secretion of Philodryas olfersii from the north-east region of Argentina.

Biocell 2003 Dec;27(3):363-70

CONICET, Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste, Sargento Cabral 2139, (3400) Corrientes, Argentina.

Philodryas olfersii is found in South America, from Amazonas to Patagonia. It is important to characterize the venom of P. olfersii, who inhabits the North-East region of Argentina, since snake venoms are known to exhibit considerable variability in composition and biological activities. In this work, mice weighing 18-20 g (n = 4 for each experimental group) were used. For the edematogenic activity mice were injected s.c. in the right foot pad with 50 microl of solutions containing different amounts of venom, whereas the left foot pad was injected with 50 microl of PBS. Two hours after injection mice were killed by cervical dislocation and both feet were cut off and weighed individually. For the myotoxic activity mice were injected i.m. with 100 microl of solutions containing 40 microg of venom. Blood samples were extracted after 1, 3, 6, 8, 10, 12, 14, 16 and 24 h of venom injection to determinate serum CPK activity and mice were sacrificed at the same time intervals to obtain the inoculated gastrocnemius muscle. They were fixed with Bouin solution and stained with Hematoxylin-Eosin. Results showed that P. olfersii venom exhibits a high edematogenic activity (MED = 0.31 microg) and a moderate myotoxic activity. Myonecrosis reached its highest level after 12 h of venom injection as shown by plasmatic CPK levels (5,401 +/- 330 IU/l) and microscopic assay. It demonstrates the potential toxicity of the venom of P. olfersii, who inhabits the North-East region of Argentina. It also reinforces the original warning concerning the potential danger of bites by colubrids.
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December 2003

Leucines 193 and 194 at the N-terminal domain of the XylS protein, the positive transcriptional regulator of the TOL meta-cleavage pathway, are involved in dimerization.

J Bacteriol 2003 May;185(10):3036-41

Department of Biochemistry and Molecular and Cellular Biology of Plants, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, E-18008 Granada, Spain.

Members of the AraC/XylS family of transcriptional regulators are usually organized in two domains: a conserved domain made up of 100 amino acids and frequently located at the C-terminal end, involved in DNA binding; and an N-terminal nonconserved domain involved in signal recognition, as is the case for regulators involved in the control of carbon metabolism (R. Tobes and J. L. Ramos, Nucleic Acids Res. 30:318-321, 2002). The XylS protein, which is extremely insoluble, controls expression of the meta-cleavage pathway for alkylbenzoate metabolism. We fused the N-terminal end of XylS to the maltose-binding protein (MBP) in vitro and found in glutaraldehyde cross-linking assays that the protein dimerized. Experiments with a chimeric N-terminal XylS linked to a 'LexA protein showed that the dimer was stabilized in the presence of alkylbenzoates. Sequence alignments with AraC and UreR allowed us to identify three residues, Leu193, Leu194, and Ile205, as potentially being involved in dimerization. Site-directed mutagenesis of XylS in which each of the above residues was replaced with Ala revealed that Leu193 and Leu194 were critical for activity and that a chimera in which LexA was linked to the N terminus of XylSLeu193Ala or XylSLeu194Ala was not functional. Dimerization of the chimeras MBP-N-XylSLeu193Ala and MBP-N-XylSLeu194Ala was not observed in cross-linking assays with glutaraldehyde.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC154087PMC
http://dx.doi.org/10.1128/JB.185.10.3036-3041.2003DOI Listing
May 2003

Residues 137 and 153 at the N terminus of the XylS protein influence the effector profile of this transcriptional regulator and the sigma factor used by RNA polymerase to stimulate transcription from its cognate promoter.

J Biol Chem 2002 Mar 21;277(9):7282-6. Epub 2001 Dec 21.

Department of Biochemistry, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Apartado de Correos 419, E-18008 Granada, Spain.

The 321-residue XylS and XylS1 proteins, encoded by the pWW0 and pWW53 plasmids respectively, differ in only 5 residues at positions 4, 53, 90, 137, and 153. As a result, the effector profile of XylS is wider than that of XylS1, and XylS mediates higher levels of transcription from its cognate-regulatable promoter than does XylS1. We generated a series of XylS-pWW0 mutants and found that the single mutants Asp-137-->Glu and His-153-->Asn exhibited an activation pattern different from that of the wild-type regulator. In the double-mutant XylSD137E,H153N the effector profile for benzoates was similar to that of XylS1. This suggests that these two residues are crucial for effector recognition and regulator activation to stimulate transcription. XylS-dependent transcription from its cognate promoter is mediated by RNA polymerase with sigma(32) or sigma(38), whereas XylS1 uses RNA polymerase with sigma(32) or sigma(70). We also found that point mutations at positions 137 and 153 of XylS led RNA polymerase to mediate transcription with sigma(70) rather than with sigma(38), as demonstrated by primer extension analysis in a sigma(70)-thermosensitive background proficient and deficient in sigma(38). This suggests that a positive transcriptional regulator can choose the RNA polymerase complex that mediates transcription from a given promoter.
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http://dx.doi.org/10.1074/jbc.M110226200DOI Listing
March 2002