José González-buitrago | University of Salamanca (original) (raw)
Papers by José González-buitrago
Advances in Experimental Medicine and Biology, 2019
Urine is a biological fluid that can be collected noninvasively in relatively large quantities wh... more Urine is a biological fluid that can be collected noninvasively in relatively large quantities which can be used for the search of biomarkers of disease, both diseases of the urological tract and systemic diseases. One of the most important aspects in proteomic studies is sample treatment before further analysis. Methods of preparation of a urine sample depend on the techniques that will be used later for separation and identification of the proteins. Also, urine preparation should be as simple as possible to increase reproducibility. Normal urine has a much diluted protein concentration with a high-salt content, which interferes with proteomic analysis. Thus, an initial step in the handling of urine sample should be to concentrate and eliminate salts. As range of protein concentrations in urine spans several orders of magnitude, effective proteomic analyses require either removal of most abundant protein or enrichment of the less abundant ones. In this chapter, we discuss the aspects related to the collection and treatment of urine for proteomic studies.
Microorganisms, 2020
Several artisanal cheeses are elaborated in European countries, being commonly curdled with renne... more Several artisanal cheeses are elaborated in European countries, being commonly curdled with rennets of animal origin. However, in some Spanish regions some cheeses of type “Torta” are elaborated using Cynara cardunculus L. rennets. Two of these cheeses, “Torta del Casar” and “Torta de Trujillo”, are elaborated in Cáceres province with ewe’s raw milk and matured over at least 60 days without starters. In this work, we identified the lactic acid bacteria present in these cheeses using MALDI-TOF MS and pheS gene analyses, which showed they belong to the species Lactobacillus curvatus, Lactobacillus diolivorans, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis and Leuconostoc mesenteroides. The pheS gene analysis also allowed the identification of the subspecies La. plantarum subsp. plantarum, La. paracasei subsp. paracasei and Le. mesenteroides subsp. jonggajibkimchii. Low similarity values were found in this gene for some currently accepted...
Toxicological Sciences, 2019
Acute kidney injury (AKI) is a serious syndrome with increasing incidence and health consequences... more Acute kidney injury (AKI) is a serious syndrome with increasing incidence and health consequences, and high mortality rate among critically ill patients. Acute kidney injury lacks a unified definition, has ambiguous semantic boundaries, and relies on defective diagnosis. This, in part, is due to the absence of biomarkers substratifying AKI patients into pathophysiological categories based on which prognosis can be assigned and clinical treatment differentiated. For instance, AKI involving acute tubular necrosis (ATN) is expected to have a worse prognosis than prerenal, purely hemodynamic AKI. However, no biomarker has been unambiguously associated with tubular cell death or is able to provide etiological distinction. We used a cell-based system to identify TCP1-eta in the culture medium as a noninvasive marker of damaged renal tubular cells. In rat models of AKI, TCP1-eta was increased in the urine co-relating with renal cortical tubule damage. When kidneys from ATN rats were perfus...
Enfermedades infecciosas y microbiologia clinica (English ed.), 2017
Continuing medical education: Methods of rapid diagnosis Fast methods of fungal and bacterial ide... more Continuing medical education: Methods of rapid diagnosis Fast methods of fungal and bacterial identification. MALDI-TOF mass spectrometry, chromogenic media ଝ
Enfermedades Infecciosas y Microbiología Clínica, 2017
La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología ... more La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología Clínica, por su rapidez y fiabilidad en la identificación de microorganismos. Sus resultados están perfectamente contrastados en la identificación de bacterias y levaduras. La identificación de micobacterias y hongos filamentosos presenta mayor complejidad, por la mayor heterogeneidad de espectros dentro de cada especie. La metodología es algo más compleja, y la ampliación del número de especies de referencia, y del número de espectros de cada especie, serán cruciales para alcanzar mayor eficacia. La identificación directa a partir de hemocultivos se ha implantado dada su aportación al manejo de pacientes graves, pero su aplicación a otras muestras es más compleja. Los medios de cultivos cromogénicos han supuesto también una aportación al diagnóstico rápido tanto en bacterias como en levaduras, ya que aceleran el diagnóstico, facilitan la detección de cultivos mixtos y permiten un diagnóstico rápido de especies resistentes.
Revista española de quimioterapia : publicación oficial de la Sociedad Española de Quimioterapia, 2013
Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of spee... more Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of speed and reliability. Conventional mycological identification has some disadvantages: it is frequently slow, reliability is sometimes low, and an extensive experience is required. The risk population for fungal infections, and therefore their clinical significance has progressively increased in recent years. 153 yeast and mould clinical isolates were analyzed by MALDI-TOF MS and conventional identification. When both methods were discrepant to the genus or species level, ITS-2 sequencing was performed. Results. The correlation in yeasts identification between conventional identification methods and MALDI-TOF MS was extremely high (99.2% to the species level and 100% to the genus level). The only discrepancy was checked by ITS-2 sequencing and confirmed the MALDI-TOF identification. The correlation in moulds identification was more heterogeneous. 68.7% of the isolates showed correlation at l...
PLoS ONE, 2010
Background: MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. So... more Background: MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. Methodology/Principal Findings: We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. Conclusions/Significance: MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles.
Pharmacogenomics, 2014
Aim: Antiretroviral treatment implies a high cost to the healthcare system. The aim of this study... more Aim: Antiretroviral treatment implies a high cost to the healthcare system. The aim of this study was to evaluate the clinical and economic impact of efavirenz (EFV) dose adjustment by monitoring plasma concentrations and pharmacogenetic analysis of the 516G>T CYP2B6 polymorphism. Materials & methods: One hundred and ninety HIV patients treated with EFV were studied. Plasma EFV concentrations were measured by HPLC with ultraviolet detection, and pharmacogenetic analysis was performed by Real Time (RT)-PCR. Results: One hundred and ninety patients initially treated with a standard dose of EFV (600 mg/day) were studied. In 31 (16.3%) patients, EFV dose was reduced. A total of 87.1% of patients were heterozygous/homozygous carriers (GT/TT). CD4+count increased while the minimum steady-state plasma concentration and adverse effects decreased significantly after dose adjustment. Considering only the dose reduction, the adjustments accounted for a saving of 43,539 €/year. Conclusion: T...
Klinische Wochenschrift, 1990
We have studied vitamin B6 status in 26 uremic patients, 18 on maintenance hemodialysis and 8 non... more We have studied vitamin B6 status in 26 uremic patients, 18 on maintenance hemodialysis and 8 nonhemodialyzed. The vitamin B6 status was estimated by an assay of erythrocyte aspartate aminotransferase and coenzyme stimulation. Hemodialyzed uremic patients were found to have vitamin B6 deficiency. Patients were treated with 150 mg pyridoxine hydrochloride daily for 4 weeks. Erythrocyte aspartate aminotransferase increased significantly in both groups of uremic patients, the increase being greater in hemodialyzed patients. In vitro pyridoxal phosphate stimulation produces an erythrocyte aspartate aminotransferase activity greater than that obtained before pyridoxine hydrochloride administration. After cessation of pyridoxine hydrochloride treatment, erythrocyte aspartate aminotransferase decreases in hemodialyzed patients, while it remains elevated in nonhemodialyzed patients. The data obtained appear to indicate that vitamin B6 administration to patients with chronic renal insufficiency must be appraised not only for correcting the deficit but also for increasing the intracellular pyridoxal phosphate concentration, which could modify the possible functional impairment at the level of apoenzymes that use pyridoxal phosphate.
Journal of Clinical Laboratory Analysis, 2002
The biological variation of anti‐TPO and anti‐Tg autoantibodies was studied in 17 clinically and ... more The biological variation of anti‐TPO and anti‐Tg autoantibodies was studied in 17 clinically and biochemically stable female patients with autoimmune thyroid disease (AITD), at regular monthly intervals over a period of 6 consecutive months. The mean and standard deviation (SD), within‐subject coefficient of variation (CV), between‐subject CV, index of individuality, reliability coefficient, and critical differences were as follows: for anti‐TPO 238 (197) U/ml, 9.2%, 81.4%, 0.11, 0.96, and 27.6%; and for anti‐Tg 1,785 (3,170) U/ml, 6.9%, 174%, 0.04, 0.99, and 22.3%. The data indicate a low within‐subject CV, and a high between‐subject CV that is particularly pronounced for anti‐Tg. The high individuality of both autoantibodies indicates that an isolated result compared to conventional population‐based reference intervals is of very little value for diagnosis. Furthermore, the near to 1 reliability coefficient for both autoantibodies correctly classifies the patient with respect to h...
Enfermedades Infecciosas y Microbiología Clínica, 2012
MALDI-TOF mass spectrometry (MS) is becoming a major resource in the Clinical Microbiology labora... more MALDI-TOF mass spectrometry (MS) is becoming a major resource in the Clinical Microbiology laboratory. Results on some groups of microorganisms are still controversial. We have studied the reliability of MALDI-TOF MS for the identification of anaerobic clinical isolates was studied compared to conventional biochemical methods, with rRNA 16S sequencing being used as a reference when discrepancies arose. A total of 126 anaerobic bacteria clinical isolates were studied by using API20A kits (bioMérieux, Marcy l'Étoile, France) and MALDI-TOF MS (Autoflex II, Bruker Daltonics, Germany), and using the data library BioTyper 2.0 (Bruker Daltonics, Germany). When discrepancies arose, or MALDI-TOF MS was not able to identify any microorganism, rRNA 16S sequencing was used as the reference standard. The biochemical method and MALDI-TOF MS agreed in identifying 60.9% of isolates at species level, and 20.3% of isolates at genus level. Among the 48 discrepancies observed, rRNA 16S sequencing supported MALDI-TOF MS identification, at species level, in 32 isolates (66.7%), and in 8 isolates (16.7%) at genus level. rRNA 16S sequencing supported biochemical identification in only two isolates (4.2%) at species level, and in 26 isolates (54.2%) at genus level. The eight isolates for which MALDI-TOF MS did not manage to identify, or the identification obtained was rejected by sequencing, belonged to species that are still not added to the BioTyper II data library. Results obtained in this study show that, overall, MALDI-TOF MS identification of anaerobic bacteria is more reliable than identification obtained by conventional biochemical methods (24% more correct identifications at species level). The number of major errors (incorrect identification at the genus level) is also 2.5-times lower. Moreover, all the major errors obtained by MALDI-TOF MS were due to the absence of some species in the data library. Thus, when data libraries are more complete, reliability differences between both methods will probably be even higher.
Cytokine, 2000
The biological variation of interleukin 6 (IL-6) and soluble interleukin 2 receptor (sIL2R), meas... more The biological variation of interleukin 6 (IL-6) and soluble interleukin 2 receptor (sIL2R), measured by automated enzyme immunoassay, in fifteen subjects studied at regular monthly intervals over a period of 6 consecutive months was measured. The mean and standard deviation (SD), within-subject CV, between-subject CV, individuality index (II) and reliability coefficient (R) were as follow: for sIL2R 571 (231) U/ml, 5.84%, 38.81%, 0.21 and 0.93; and for IL-6 1.43 (0.9) pg/ml, 48.48%, 39.38%, 1.44, and 0.37. The data indicate a relatively high between-subject CV, quite similar in both cases, and a within-subject CV much higher for IL-6 than for sIL2R. Thus, reference values can be used for diagnosis for IL6 (high II), while not for sIL2R (low II). However, the low R for IL-6 implies that more than one measurement are needed. sIL2R has a very high R and a relatively small critical differences, a circumstance appropriate for follow-up.
Clinica Chimica Acta, 2007
Allergic diseases are (IgE)-mediated hypersensitivity reactions affecting more than 25% of the wo... more Allergic diseases are (IgE)-mediated hypersensitivity reactions affecting more than 25% of the world's population. Proteomic technologies have been increasingly used in the field of allergy and include the use of protein microarrays and two-dimensional gel electrophoresis coupled with immunoblotting. The literature relevant to proteomic approaches to allergic diseases was searched using MEDLINE database. We reviewed proteomics approaches and applications, focusing specifically on two-dimensional immunoblotting techniques and allergen microarrays. The results obtained show that proteomic approaches using two-dimensional immunoblotting appear to be a powerful strategy for the identification of allergenic proteins. Likewise, the use of allergen microarrays allows a large number of IgE antibodies to be simultaneously identified. Proteomic approaches are only beginning to be applied to the study of allergy. In the field of in vitro diagnosis, allergen microarrays provide a promising tool not routinely used in the allergy laboratory. In the near future this powerful technique will be used as a standard technique for in vitro diagnosis of allergy.
Electrophoresis, Sep 1, 2013
This article has been accepted for publication and undergone full peer review but has not been th... more This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as
Urologia Internationalis, 1985
The content of cytoplasmic androgen receptors (Bmax) was analyzed in non-tumoral renal tissue of ... more The content of cytoplasmic androgen receptors (Bmax) was analyzed in non-tumoral renal tissue of 12 men and 15 postmenopausal women using a synthetic androgen (methyltrienolone) as ligand and a method of dextran-coated charcoal. The Bmax in both sexes was compared, establishing correlations between it and circulating levels of testosterone, dehydroepiandrosterone sulfate, androstenedione, and estradiol to find out the possible influence of the hormonal environment on androgen receptors in the human kidney. No differences in Bmax were observed between males (46.3 +/- 24 fmol/g of tissue) and females (45.4 +/- 26 fmol/g), in spite of the significantly greater (p less than 0.01) levels of circulating testosterone in the former group. No significant linear correlations existed between any of the steroids analyzed and the Bmax. These results demonstrate the existence of androgenic receptors in non-tumoral human kidney and indicate that its content is not regulated by circulating levels of testosterone. The concentrations of the principal extratesticular androgens and estradiol do not seem to have a quantitative influence on these androphilic proteins either.
Stereotactic and Functional Neurosurgery, 1990
It is presumed today that spinal cord stimulation induces local delivery of vasoactive substances... more It is presumed today that spinal cord stimulation induces local delivery of vasoactive substances, such as prostacyclins, histamine, substance P, and vasoactive neuropeptides, in the perivascular environment and the vascular wall to mediate the segmental vasodilator response. To investigate this mechanism, 9 dogs were subjected to low thoracic spinal cord stimulation. Venous and arterial blood samples from the paraesthesic area in the lower limbs were obtained before and 120 min after stimulation to measure changes in the plasma concentration of vasoactive intestinal peptide, substance P, and histamine. The results were compared with those obtained from vessels of the upper limbs. Blood flow changes following stimulation were recorded by electromagnetic flowmeters. Local arterial vasoactive intestinal peptide showed a mean increase of 33% after 60 min of stimulation. Changes concerning substance P were inconclusive. Local arterial and venous histamine concentrations increased 26 and 29%, respectively, after 60 min of stimulation.
Revista española de fisiología, 1988
Enzyme levels of lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), asparta... more Enzyme levels of lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured in the cytosol of renal cortex samples from either normal and pathologic kidney tissue. The mean enzyme activity values, expressed in Units per gram of cytosolic protein decreased in the following order: normal cortex (LDH = 4,299 +/- 654; AST = 522 +/- 101; ALT = 197 +/- 44). chronic pyelonephritis (LDH = 2,360 +/- 876; AST = 297 +/- 117; ALT = 90 +/- 48), hydronephrosis (LDH = 2,208 +/- 1,264; AST = 279 +/- 165; ALT = 82 +/- 61), pyonephrosis (LDH = 1,410 +/- 596; AST = 158 +/- 69; ALT = 23.4 +/- 16.4) and renal tuberculosis (LDH = 1,149 +/- 481; AST = 93 +/- 34; ALT = 5.6 +/- 2.8). The decrease in the enzyme activities paralleled tissue damage and it was shown to affect cellular functionality in relation with energy and amino acid metabolism.
Anticancer research
We evaluated the potential role of serial preinstillation levels of several interleukins, TNFalph... more We evaluated the potential role of serial preinstillation levels of several interleukins, TNFalpha and urinary tumor markers to monitor patients with bladder cancer receiving intravesical BCG. 121 urine samples were collected from: patients with bladder cancer treated with BCG (group 1); patients with bladder cancer receiving other intravesical treatment (group 2) and patients with urinary tract infections (group 3). Cytokines [IL-2, IL6 and [L8] and TNFalpha and urinary tumor markers [UBC, CYFRA 21-1 and NMP22] were measured by immunoassays. In 3 out of 15 BCG non-responders that recurred over the period of the study, no cytokine peak for IL-2, IL-6 or TNFa were detected. Urinary tumor markers increased in 2 out of 3 of these patients earlier than scheduled cystoscopies. Cytokine measurement was heterogeneous among 12 out of 15 BCG-responding patients: there were low levels of IL-6 and TNFalpha and peaks of IL-2 and IL-8 in 10 out of 12 and 4 out of 12 patients, respectively. Durin...
The Journal of pharmacology and experimental therapeutics, 1995
This study examines the ability of S-adenosyl-L-methionine to prevent or antagonize the cyclospor... more This study examines the ability of S-adenosyl-L-methionine to prevent or antagonize the cyclosporine A-induced adverse effects on biliary secretion in the rat. S-adenosyl-L-methionine was administered as a single bolus 3, 5 and 8 hr before administering a single dose of cyclosporine A, and also concurrently administered with cyclosporine A for 1 or 2 wk. Acute S-adenosyl-L-methionine preadministration attenuated the cyclosporine-induced cholestasis and inhibition of bile acids, cholesterol and phospholipid biliary secretion. S-adenosyl-L-methionine pretreatment for 1 wk and simultaneous cotreatment with cyclosporine for 1 or 2 wk not only maintained the beneficial effects reported above but further improved them because the adverse effects of the immunosuppressor drug were prevented or antagonized by S-adenosyl-L-methionine. These results provide the first direct evidence of the ability of exogenously administered S-adenosyl-L-methionine to antagonize cyclosporine-induced abnormalit...
Advances in Experimental Medicine and Biology, 2019
Urine is a biological fluid that can be collected noninvasively in relatively large quantities wh... more Urine is a biological fluid that can be collected noninvasively in relatively large quantities which can be used for the search of biomarkers of disease, both diseases of the urological tract and systemic diseases. One of the most important aspects in proteomic studies is sample treatment before further analysis. Methods of preparation of a urine sample depend on the techniques that will be used later for separation and identification of the proteins. Also, urine preparation should be as simple as possible to increase reproducibility. Normal urine has a much diluted protein concentration with a high-salt content, which interferes with proteomic analysis. Thus, an initial step in the handling of urine sample should be to concentrate and eliminate salts. As range of protein concentrations in urine spans several orders of magnitude, effective proteomic analyses require either removal of most abundant protein or enrichment of the less abundant ones. In this chapter, we discuss the aspects related to the collection and treatment of urine for proteomic studies.
Microorganisms, 2020
Several artisanal cheeses are elaborated in European countries, being commonly curdled with renne... more Several artisanal cheeses are elaborated in European countries, being commonly curdled with rennets of animal origin. However, in some Spanish regions some cheeses of type “Torta” are elaborated using Cynara cardunculus L. rennets. Two of these cheeses, “Torta del Casar” and “Torta de Trujillo”, are elaborated in Cáceres province with ewe’s raw milk and matured over at least 60 days without starters. In this work, we identified the lactic acid bacteria present in these cheeses using MALDI-TOF MS and pheS gene analyses, which showed they belong to the species Lactobacillus curvatus, Lactobacillus diolivorans, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis and Leuconostoc mesenteroides. The pheS gene analysis also allowed the identification of the subspecies La. plantarum subsp. plantarum, La. paracasei subsp. paracasei and Le. mesenteroides subsp. jonggajibkimchii. Low similarity values were found in this gene for some currently accepted...
Toxicological Sciences, 2019
Acute kidney injury (AKI) is a serious syndrome with increasing incidence and health consequences... more Acute kidney injury (AKI) is a serious syndrome with increasing incidence and health consequences, and high mortality rate among critically ill patients. Acute kidney injury lacks a unified definition, has ambiguous semantic boundaries, and relies on defective diagnosis. This, in part, is due to the absence of biomarkers substratifying AKI patients into pathophysiological categories based on which prognosis can be assigned and clinical treatment differentiated. For instance, AKI involving acute tubular necrosis (ATN) is expected to have a worse prognosis than prerenal, purely hemodynamic AKI. However, no biomarker has been unambiguously associated with tubular cell death or is able to provide etiological distinction. We used a cell-based system to identify TCP1-eta in the culture medium as a noninvasive marker of damaged renal tubular cells. In rat models of AKI, TCP1-eta was increased in the urine co-relating with renal cortical tubule damage. When kidneys from ATN rats were perfus...
Enfermedades infecciosas y microbiologia clinica (English ed.), 2017
Continuing medical education: Methods of rapid diagnosis Fast methods of fungal and bacterial ide... more Continuing medical education: Methods of rapid diagnosis Fast methods of fungal and bacterial identification. MALDI-TOF mass spectrometry, chromogenic media ଝ
Enfermedades Infecciosas y Microbiología Clínica, 2017
La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología ... more La espectrometría de masas MALDI-TOF es ya una herramienta de trabajo rutinaria en Microbiología Clínica, por su rapidez y fiabilidad en la identificación de microorganismos. Sus resultados están perfectamente contrastados en la identificación de bacterias y levaduras. La identificación de micobacterias y hongos filamentosos presenta mayor complejidad, por la mayor heterogeneidad de espectros dentro de cada especie. La metodología es algo más compleja, y la ampliación del número de especies de referencia, y del número de espectros de cada especie, serán cruciales para alcanzar mayor eficacia. La identificación directa a partir de hemocultivos se ha implantado dada su aportación al manejo de pacientes graves, pero su aplicación a otras muestras es más compleja. Los medios de cultivos cromogénicos han supuesto también una aportación al diagnóstico rápido tanto en bacterias como en levaduras, ya que aceleran el diagnóstico, facilitan la detección de cultivos mixtos y permiten un diagnóstico rápido de especies resistentes.
Revista española de quimioterapia : publicación oficial de la Sociedad Española de Quimioterapia, 2013
Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of spee... more Recently, bacterial identification by MALDI-TOF MS has acquired a high relevance in terms of speed and reliability. Conventional mycological identification has some disadvantages: it is frequently slow, reliability is sometimes low, and an extensive experience is required. The risk population for fungal infections, and therefore their clinical significance has progressively increased in recent years. 153 yeast and mould clinical isolates were analyzed by MALDI-TOF MS and conventional identification. When both methods were discrepant to the genus or species level, ITS-2 sequencing was performed. Results. The correlation in yeasts identification between conventional identification methods and MALDI-TOF MS was extremely high (99.2% to the species level and 100% to the genus level). The only discrepancy was checked by ITS-2 sequencing and confirmed the MALDI-TOF identification. The correlation in moulds identification was more heterogeneous. 68.7% of the isolates showed correlation at l...
PLoS ONE, 2010
Background: MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. So... more Background: MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. Methodology/Principal Findings: We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. Conclusions/Significance: MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles.
Pharmacogenomics, 2014
Aim: Antiretroviral treatment implies a high cost to the healthcare system. The aim of this study... more Aim: Antiretroviral treatment implies a high cost to the healthcare system. The aim of this study was to evaluate the clinical and economic impact of efavirenz (EFV) dose adjustment by monitoring plasma concentrations and pharmacogenetic analysis of the 516G>T CYP2B6 polymorphism. Materials & methods: One hundred and ninety HIV patients treated with EFV were studied. Plasma EFV concentrations were measured by HPLC with ultraviolet detection, and pharmacogenetic analysis was performed by Real Time (RT)-PCR. Results: One hundred and ninety patients initially treated with a standard dose of EFV (600 mg/day) were studied. In 31 (16.3%) patients, EFV dose was reduced. A total of 87.1% of patients were heterozygous/homozygous carriers (GT/TT). CD4+count increased while the minimum steady-state plasma concentration and adverse effects decreased significantly after dose adjustment. Considering only the dose reduction, the adjustments accounted for a saving of 43,539 €/year. Conclusion: T...
Klinische Wochenschrift, 1990
We have studied vitamin B6 status in 26 uremic patients, 18 on maintenance hemodialysis and 8 non... more We have studied vitamin B6 status in 26 uremic patients, 18 on maintenance hemodialysis and 8 nonhemodialyzed. The vitamin B6 status was estimated by an assay of erythrocyte aspartate aminotransferase and coenzyme stimulation. Hemodialyzed uremic patients were found to have vitamin B6 deficiency. Patients were treated with 150 mg pyridoxine hydrochloride daily for 4 weeks. Erythrocyte aspartate aminotransferase increased significantly in both groups of uremic patients, the increase being greater in hemodialyzed patients. In vitro pyridoxal phosphate stimulation produces an erythrocyte aspartate aminotransferase activity greater than that obtained before pyridoxine hydrochloride administration. After cessation of pyridoxine hydrochloride treatment, erythrocyte aspartate aminotransferase decreases in hemodialyzed patients, while it remains elevated in nonhemodialyzed patients. The data obtained appear to indicate that vitamin B6 administration to patients with chronic renal insufficiency must be appraised not only for correcting the deficit but also for increasing the intracellular pyridoxal phosphate concentration, which could modify the possible functional impairment at the level of apoenzymes that use pyridoxal phosphate.
Journal of Clinical Laboratory Analysis, 2002
The biological variation of anti‐TPO and anti‐Tg autoantibodies was studied in 17 clinically and ... more The biological variation of anti‐TPO and anti‐Tg autoantibodies was studied in 17 clinically and biochemically stable female patients with autoimmune thyroid disease (AITD), at regular monthly intervals over a period of 6 consecutive months. The mean and standard deviation (SD), within‐subject coefficient of variation (CV), between‐subject CV, index of individuality, reliability coefficient, and critical differences were as follows: for anti‐TPO 238 (197) U/ml, 9.2%, 81.4%, 0.11, 0.96, and 27.6%; and for anti‐Tg 1,785 (3,170) U/ml, 6.9%, 174%, 0.04, 0.99, and 22.3%. The data indicate a low within‐subject CV, and a high between‐subject CV that is particularly pronounced for anti‐Tg. The high individuality of both autoantibodies indicates that an isolated result compared to conventional population‐based reference intervals is of very little value for diagnosis. Furthermore, the near to 1 reliability coefficient for both autoantibodies correctly classifies the patient with respect to h...
Enfermedades Infecciosas y Microbiología Clínica, 2012
MALDI-TOF mass spectrometry (MS) is becoming a major resource in the Clinical Microbiology labora... more MALDI-TOF mass spectrometry (MS) is becoming a major resource in the Clinical Microbiology laboratory. Results on some groups of microorganisms are still controversial. We have studied the reliability of MALDI-TOF MS for the identification of anaerobic clinical isolates was studied compared to conventional biochemical methods, with rRNA 16S sequencing being used as a reference when discrepancies arose. A total of 126 anaerobic bacteria clinical isolates were studied by using API20A kits (bioMérieux, Marcy l'Étoile, France) and MALDI-TOF MS (Autoflex II, Bruker Daltonics, Germany), and using the data library BioTyper 2.0 (Bruker Daltonics, Germany). When discrepancies arose, or MALDI-TOF MS was not able to identify any microorganism, rRNA 16S sequencing was used as the reference standard. The biochemical method and MALDI-TOF MS agreed in identifying 60.9% of isolates at species level, and 20.3% of isolates at genus level. Among the 48 discrepancies observed, rRNA 16S sequencing supported MALDI-TOF MS identification, at species level, in 32 isolates (66.7%), and in 8 isolates (16.7%) at genus level. rRNA 16S sequencing supported biochemical identification in only two isolates (4.2%) at species level, and in 26 isolates (54.2%) at genus level. The eight isolates for which MALDI-TOF MS did not manage to identify, or the identification obtained was rejected by sequencing, belonged to species that are still not added to the BioTyper II data library. Results obtained in this study show that, overall, MALDI-TOF MS identification of anaerobic bacteria is more reliable than identification obtained by conventional biochemical methods (24% more correct identifications at species level). The number of major errors (incorrect identification at the genus level) is also 2.5-times lower. Moreover, all the major errors obtained by MALDI-TOF MS were due to the absence of some species in the data library. Thus, when data libraries are more complete, reliability differences between both methods will probably be even higher.
Cytokine, 2000
The biological variation of interleukin 6 (IL-6) and soluble interleukin 2 receptor (sIL2R), meas... more The biological variation of interleukin 6 (IL-6) and soluble interleukin 2 receptor (sIL2R), measured by automated enzyme immunoassay, in fifteen subjects studied at regular monthly intervals over a period of 6 consecutive months was measured. The mean and standard deviation (SD), within-subject CV, between-subject CV, individuality index (II) and reliability coefficient (R) were as follow: for sIL2R 571 (231) U/ml, 5.84%, 38.81%, 0.21 and 0.93; and for IL-6 1.43 (0.9) pg/ml, 48.48%, 39.38%, 1.44, and 0.37. The data indicate a relatively high between-subject CV, quite similar in both cases, and a within-subject CV much higher for IL-6 than for sIL2R. Thus, reference values can be used for diagnosis for IL6 (high II), while not for sIL2R (low II). However, the low R for IL-6 implies that more than one measurement are needed. sIL2R has a very high R and a relatively small critical differences, a circumstance appropriate for follow-up.
Clinica Chimica Acta, 2007
Allergic diseases are (IgE)-mediated hypersensitivity reactions affecting more than 25% of the wo... more Allergic diseases are (IgE)-mediated hypersensitivity reactions affecting more than 25% of the world's population. Proteomic technologies have been increasingly used in the field of allergy and include the use of protein microarrays and two-dimensional gel electrophoresis coupled with immunoblotting. The literature relevant to proteomic approaches to allergic diseases was searched using MEDLINE database. We reviewed proteomics approaches and applications, focusing specifically on two-dimensional immunoblotting techniques and allergen microarrays. The results obtained show that proteomic approaches using two-dimensional immunoblotting appear to be a powerful strategy for the identification of allergenic proteins. Likewise, the use of allergen microarrays allows a large number of IgE antibodies to be simultaneously identified. Proteomic approaches are only beginning to be applied to the study of allergy. In the field of in vitro diagnosis, allergen microarrays provide a promising tool not routinely used in the allergy laboratory. In the near future this powerful technique will be used as a standard technique for in vitro diagnosis of allergy.
Electrophoresis, Sep 1, 2013
This article has been accepted for publication and undergone full peer review but has not been th... more This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as
Urologia Internationalis, 1985
The content of cytoplasmic androgen receptors (Bmax) was analyzed in non-tumoral renal tissue of ... more The content of cytoplasmic androgen receptors (Bmax) was analyzed in non-tumoral renal tissue of 12 men and 15 postmenopausal women using a synthetic androgen (methyltrienolone) as ligand and a method of dextran-coated charcoal. The Bmax in both sexes was compared, establishing correlations between it and circulating levels of testosterone, dehydroepiandrosterone sulfate, androstenedione, and estradiol to find out the possible influence of the hormonal environment on androgen receptors in the human kidney. No differences in Bmax were observed between males (46.3 +/- 24 fmol/g of tissue) and females (45.4 +/- 26 fmol/g), in spite of the significantly greater (p less than 0.01) levels of circulating testosterone in the former group. No significant linear correlations existed between any of the steroids analyzed and the Bmax. These results demonstrate the existence of androgenic receptors in non-tumoral human kidney and indicate that its content is not regulated by circulating levels of testosterone. The concentrations of the principal extratesticular androgens and estradiol do not seem to have a quantitative influence on these androphilic proteins either.
Stereotactic and Functional Neurosurgery, 1990
It is presumed today that spinal cord stimulation induces local delivery of vasoactive substances... more It is presumed today that spinal cord stimulation induces local delivery of vasoactive substances, such as prostacyclins, histamine, substance P, and vasoactive neuropeptides, in the perivascular environment and the vascular wall to mediate the segmental vasodilator response. To investigate this mechanism, 9 dogs were subjected to low thoracic spinal cord stimulation. Venous and arterial blood samples from the paraesthesic area in the lower limbs were obtained before and 120 min after stimulation to measure changes in the plasma concentration of vasoactive intestinal peptide, substance P, and histamine. The results were compared with those obtained from vessels of the upper limbs. Blood flow changes following stimulation were recorded by electromagnetic flowmeters. Local arterial vasoactive intestinal peptide showed a mean increase of 33% after 60 min of stimulation. Changes concerning substance P were inconclusive. Local arterial and venous histamine concentrations increased 26 and 29%, respectively, after 60 min of stimulation.
Revista española de fisiología, 1988
Enzyme levels of lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), asparta... more Enzyme levels of lactate dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (HBDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured in the cytosol of renal cortex samples from either normal and pathologic kidney tissue. The mean enzyme activity values, expressed in Units per gram of cytosolic protein decreased in the following order: normal cortex (LDH = 4,299 +/- 654; AST = 522 +/- 101; ALT = 197 +/- 44). chronic pyelonephritis (LDH = 2,360 +/- 876; AST = 297 +/- 117; ALT = 90 +/- 48), hydronephrosis (LDH = 2,208 +/- 1,264; AST = 279 +/- 165; ALT = 82 +/- 61), pyonephrosis (LDH = 1,410 +/- 596; AST = 158 +/- 69; ALT = 23.4 +/- 16.4) and renal tuberculosis (LDH = 1,149 +/- 481; AST = 93 +/- 34; ALT = 5.6 +/- 2.8). The decrease in the enzyme activities paralleled tissue damage and it was shown to affect cellular functionality in relation with energy and amino acid metabolism.
Anticancer research
We evaluated the potential role of serial preinstillation levels of several interleukins, TNFalph... more We evaluated the potential role of serial preinstillation levels of several interleukins, TNFalpha and urinary tumor markers to monitor patients with bladder cancer receiving intravesical BCG. 121 urine samples were collected from: patients with bladder cancer treated with BCG (group 1); patients with bladder cancer receiving other intravesical treatment (group 2) and patients with urinary tract infections (group 3). Cytokines [IL-2, IL6 and [L8] and TNFalpha and urinary tumor markers [UBC, CYFRA 21-1 and NMP22] were measured by immunoassays. In 3 out of 15 BCG non-responders that recurred over the period of the study, no cytokine peak for IL-2, IL-6 or TNFa were detected. Urinary tumor markers increased in 2 out of 3 of these patients earlier than scheduled cystoscopies. Cytokine measurement was heterogeneous among 12 out of 15 BCG-responding patients: there were low levels of IL-6 and TNFalpha and peaks of IL-2 and IL-8 in 10 out of 12 and 4 out of 12 patients, respectively. Durin...
The Journal of pharmacology and experimental therapeutics, 1995
This study examines the ability of S-adenosyl-L-methionine to prevent or antagonize the cyclospor... more This study examines the ability of S-adenosyl-L-methionine to prevent or antagonize the cyclosporine A-induced adverse effects on biliary secretion in the rat. S-adenosyl-L-methionine was administered as a single bolus 3, 5 and 8 hr before administering a single dose of cyclosporine A, and also concurrently administered with cyclosporine A for 1 or 2 wk. Acute S-adenosyl-L-methionine preadministration attenuated the cyclosporine-induced cholestasis and inhibition of bile acids, cholesterol and phospholipid biliary secretion. S-adenosyl-L-methionine pretreatment for 1 wk and simultaneous cotreatment with cyclosporine for 1 or 2 wk not only maintained the beneficial effects reported above but further improved them because the adverse effects of the immunosuppressor drug were prevented or antagonized by S-adenosyl-L-methionine. These results provide the first direct evidence of the ability of exogenously administered S-adenosyl-L-methionine to antagonize cyclosporine-induced abnormalit...