Domestication influences morphological and physiological responses to salinity in Brassica oleracea seedlings.

Brassica oleracea cultivars include important vegetable and forage crops grown worldwide, whereas the wild counterpart occurs naturally on European sea cliffs. Domestication and selection processes have led to phenotypic and genetic divergence between domesticated plants and their wild ancestors that inhabit coastal areas and are exposed to saline conditions. Salinity is one of the most limiting factors for crop production. However, little is known about how salinity affects plants in relation to domestication of B. oleracea. The objective of this study was to determine the influence of domestication status (wild, landrace or cultivar) on the response of different B. oleracea crops to salinity, as measured by seed germination, plant growth, water content and mineral concentration parameters at the seedling stage. For this purpose, two independent pot experiments were conducted with six accessions of B. oleracea, including cabbage (group capitata) and kale (group acephala), in a growth chamber under controlled environmental conditions. In both taxonomic groups, differences in domestication status and salt stress significantly affected all major process such as germination, changes in dry matter, water relations and mineral uptake. In the acephala experiment, the domestication × salinity interaction significantly affected water content parameters and shoot Na+ allocation. At early stages of development, wild plants are more succulent than cultivated plants and have a higher capacity to maintain lower Na+ concentrations in their shoots in response to increasing levels of salinity. Different responses of domesticated and cultivated accessions in relation to these traits indicated a high level of natural variation in wild B. oleracea. Exclusion of Na+ from shoots and increasing succulence may enhance salt tolerance in B. oleracea exposed to extreme salinity in the long term. The wild germplasm can potentially be used to improve the salt tolerance of crops by the identification of useful genes and incorporation of these into salinity-sensitive cultivars.

CALL ME BY YOUR NAME. Paciente protegido: reflexiones sobre lo que implica privar del nombre propio para la Seguridad del Paciente / CALL ME BY YOUR NAME. Protected patient: reflections on removing the patient's name in Patient Safety

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Development of a construct-based risk assessment framework for genetic engineered crops.

Experience gained in the risk assessment (RA) of genetically engineered (GE) crops since their first experimental introductions in the early nineties, has increased the level of familiarity with these breeding methodologies and has motivated several agencies and expert groups worldwide to revisit the scientific criteria underlying the RA process. Along these lines, the need to engage in a scientific discussion for the case of GE crops transformed with similar constructs was recently identified in Argentina. In response to this need, the Argentine branch of the International Life Sciences Institute (ILSI Argentina) convened a tripartite working group to discuss a science-based evaluation approach for transformation events developed with genetic constructs which are identical or similar to those used in previously evaluated or approved GE crops. This discussion considered new transformation events within the same or different species and covered both environmental and food safety aspects. A construct similarity concept was defined, considering the biological function of the introduced genes. Factors like environmental and dietary exposure, familiarity with both the crop and the trait as well as the crop biology, were identified as key to inform a construct-based RA process.

In vitro activity of glucosinolates and their degradation products against brassica-pathogenic bacteria and fungi.

Glucosinolates (GSLs) are secondary metabolites found in Brassica vegetables that confer on them resistance against pests and diseases. Both GSLs and glucosinolate hydrolysis products (GHPs) have shown positive effects in reducing soil pathogens. Information about their in vitro biocide effects is scarce, but previous studies have shown sinigrin GSLs and their associated allyl isothiocyanate (AITC) to be soil biocides. The objective of this work was to evaluate the biocide effects of 17 GSLs and GHPs and of leaf methanolic extracts of different GSL-enriched Brassica crops on suppressing in vitro growth of two bacterial (Xanthomonas campestris pv. campestris and Pseudomonas syringae pv. maculicola) and two fungal (Alternaria brassicae and Sclerotinia scletoriorum) Brassica pathogens. GSLs, GHPs, and methanolic leaf extracts inhibited the development of the pathogens tested compared to the control, and the effect was dose dependent. Furthermore, the biocide effects of the different compounds studied were dependent on the species and race of the pathogen. These results indicate that GSLs and their GHPs, as well as extracts of different Brassica species, have potential to inhibit pathogen growth and offer new opportunities to study the use of Brassica crops in biofumigation for the control of multiple diseases.

Monounsaturated fatty acid aerobic synthesis in Bradyrhizobium TAL1000 peanut-nodulating is affected by temperature.

AIMS: The aim of this work was to clarify the mechanism of monounsaturated fatty acid (MUFA) synthesis in Bradyrhizobium TAL1000 and the effect of high temperature on this process. METHODS AND RESULTS: Bradyrhizobium TAL1000 was exposed to a high growth temperature and heat shock, and fatty acid composition and synthesis were tested. To determine the presence of a possible desaturase, a gene was identify and overexpressed in Escherichia coli. The desaturase expression was detected by RT-PCR and Western blotting. In B. TAL1000, an aerobic mechanism for MUFA synthesis was detected. Desaturation was decreased by high growth temperature and by heat shock. Two hours of exposure to 37°C were required for the change in MUFA levels. A potential ∆9 desaturase gene was identified and successfully expressed in E. coli. A high growth temperature and not heat shock reduced transcript and protein desaturase levels in rhizobial strain. CONCLUSIONS: In B. TAL1000, the anaerobic MUFA biosynthetic pathway is supplemented by an aerobic mechanism mediated by desaturase and is down-regulated by temperature to maintain membrane fluidity under stressful conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This knowledge will be useful for developing strategies to improve a sustainable practice of this bacterium under stress and to enhance the bioprocess for the inoculants' manufacture.

Identification of Sources of Resistance to Xanthomonas campestris pv. campestris in Brassica napus Crops.

Black rot, caused by Xanthomonas campestris pv. campestris, is one of the most important diseases affecting Brassica crops worldwide. Nine races have been differentiated in X. campestris pv. campestris, with races 1 and 4 being the most virulent and widespread. The objective of this work was to identify sources of resistance to races 1 and 4 of X. campestris pv. campestris in different Brassica napus crops, mainly in the underexplored pabularia group. Seventy-six accessions belonging to four B. napus groups were screened for resistance to two X. campestris pv. campestris races (1 and 4). The strain of race 1 used in this study was more virulent on the tested materials than the strain of race 4. No race-specific resistance was found to race 1. Most cultivars were susceptible except Russian kale, from the pabularia group, which showed some resistant plants and some other accessions with some partially resistant plants. High levels of race-specific resistance to race 4 were found in the pabularia group, and great variability within accessions was identified. Three improved cultivars (Ragged Jack kale, Friese Gele, and Valle del Oro) and four landraces (Russian kale, MBG-BRS0037, MBG-BRS0041, and MBG-BRS0131) showed plants with some degree of resistance to both races, which may indicate that race-nonspecific resistance is involved. These accessions could be directly used in breeding programs, either as improved cultivars or as donors of race-specific resistance to other Brassica cultivars.

When does acute pain become chronic?

The transition from acute to chronic pain appears to occur in discrete pathophysiological and histopathological steps. Stimuli initiating a nociceptive response vary, but receptors and endogenous defence mechanisms in the periphery interact in a similar manner regardless of the insult. Chemical, mechanical, and thermal receptors, along with leucocytes and macrophages, determine the intensity, location, and duration of noxious events. Noxious stimuli are transduced to the dorsal horn of the spinal cord, where amino acid and peptide transmitters activate second-order neurones. Spinal neurones then transmit signals to the brain. The resultant actions by the individual involve sensory-discriminative, motivational-affective, and modulatory processes in an attempt to limit or stop the painful process. Under normal conditions, noxious stimuli diminish as healing progresses and pain sensation lessens until minimal or no pain is detected. Persistent, intense pain, however, activates secondary mechanisms both at the periphery and within the central nervous system that cause allodynia, hyperalgesia, and hyperpathia that can diminish normal functioning. These changes begin in the periphery with upregulation of cyclo-oxygenase-2 and interleukin-1ß-sensitizing first-order neurones, which eventually sensitize second-order spinal neurones by activating N-methyl-d-aspartic acid channels and signalling microglia to alter neuronal cytoarchitecture. Throughout these processes, prostaglandins, endocannabinoids, ion-specific channels, and scavenger cells all play a key role in the transformation of acute to chronic pain. A better understanding of the interplay among these substances will assist in the development of agents designed to ameliorate or reverse chronic pain.

Resistance of cabbage (Brassica oleracea capitata group) crops to Mamestra brassicae.

Twenty-one cabbage (Brassica oleracea capitata group) varieties, including 16 local varieties and five commercial hybrids, were screened for resistance to the moth Mamestra brassicae L. under natural and artificial conditions in northwestern Spain. Resistance was assessed as the proportion of damaged plants and damaged leaves, leaf feeding injury, and number of larvae present. Correlation coefficients among damage traits showed that a visual scale (general appearance rating) should be a useful indicator of resistance. Most local varieties were highly susceptible to M. brassicae, whereas the commercial hybrids tested were resistant in terms of head foliage consumption and number of larvae per plant. Performance of varieties was similar under natural and artificial infestation although some of them performed differently at each year. Three local varieties (MBG-BRS0057, MBG-BRS0074, and MBG-BRS0452) were highly susceptible at both natural and artificial infestation conditions being MBG-BRS0074 the most damaged variety. Two local varieties (MBG-BRS0402 and MBG-BRS0535) and commercial hybrids were identified as resistant or moderately resistant to M. brassicae. Among them, 'Corazón de Buey' and 'Cabeza negra' were the most resistant and produced compact heads. These varieties could be useful sources of resistance to obtain resistant varieties to M. brassicae or as donors of resistance to other Brassica crops. The possible role of leaf traits, head compactness, and leaf glucosinolate content in relation to M. brassicae resistance is discussed.

Performance of automated slidemakers and stainers in a working laboratory environment - routine operation and quality control.

The automated slidemaker/stainers of the four Beckman Coulter LH755 hematology systems in our laboratory are operated as analyzers, with similar requirements for setup, maintenance and quality control. A study was performed to confirm that these slide maker/stainers in routine use produce peripheral blood films that are completely satisfactory for microscopy and without cells, particularly abnormal cells, being pulled to the edges or sides of the film outside the usual working area. One hundred and thirty-nine automated blood films that had been produced during routine operation were compared with well-prepared manual films from the same patients. None of the films was unacceptable for microscopy. The distributions of normal white cell types within the counting areas of automated films compared with manual films, for all 139 samples for WBC from 1.0 to 352.8 x 10(9)/l; for blasts and promyelocytes in the 65 samples in which they occurred and for nucleated red blood cells in the 58 samples in which they occurred all fell within the expected limits of 200 cell differential counts of CLSI H20-A. Red cell morphology and the occurrence of WBC clumps, platelet clumps and smudge cells were comparable between the automated and manual films of all samples. We conclude that automated slidemaker/stainers, as typified by those of the Beckman Coulter LH755 system, are capable of producing blood films comparable with well-prepared manual films in routine laboratory use; and that the maintenance and quality control procedures used in our laboratory ensure consistent high quality performance from these systems.

Comprehensive lung injury pathology induced by mTOR inhibitors

Interstitial lung disease is a rare side effect of temsirolimus treatment in renal cancer patients. Pulmonary fibrosis is characterised by the accumulation of extracellular matrix collagen, fibroblast proliferation and migration, and loss of alveolar gas exchange units. Previous studies of pulmonary fibrosis have mainly focused on the fibroproliferative process in the lungs. However, the molecular mechanism by which sirolimus promotes lung fibrosis remains elusive. Here, we propose an overall cascade hypothesis of interstitial lung diseases that represents a common, partly underlying synergism among them as well as the lung pathogenesis side effects of mammalian target of rapamycin inhibitors (AU)

Short daily versus conventional hemodialysis quality of life: a cross-sectional multicentric study in Spain.

BACKGROUND: End-stage renal disease is a major health problem worldwide nowadays. Although conventional hemodialysis is the most widely used modality, short daily hemodialysis has been proposed as a more physiologic treatment. The objective of this article is to compare the quality of life of patients on each hemodialysis modality. METHODS: A multicentric cross-sectional study was performed in 9 Spanish hospitals. Patients treated for at least 3 months with conventional or short daily hemodialysis were included and quality of life measured using the Euroqol-5D quality of life questionnaire. Bayesian models were used for analyzing quality of life results. RESULTS: Ninety-three patients were included, 27 were on daily hemodialysis and 66 on conventional hemodialysis. All models demonstrated a better quality of life for daily hemodialysis versus conventional hemodialysis. Only 14% of the patients on conventional hemodialysis were willing to change to a daily schedule. CONCLUSIONS: Short daily hemodialysis shows a better quality of life than conventional hemodialysis with all Bayesian approaches considered.

[Clinical course of multiple sclerosis in patients treated with cytostatic drugs for cancer]. / Evolución clínica de pacientes con esclerosis múltiple tratados con citostáticos por cáncer.

INTRODUCTION: The risk of side effects secondary to global and non-specific immune suppression has limited the systematic application of immunosuppressive therapy in multiple sclerosis (MS). However, when a patient with MS develops a cancer, cytostatic drugs as treatment for the neoplastic process may induce improvement not only of the cancer but also of MS. CASE REPORTS: We present a series of four women with clinically defined MS and subjected to cytostatic therapy (cisplatin, 5-fluorouracil, leukovorin, adriamycin, tamoxifen and anastrozole) after the development of cancer: two presented breast cancer, one colon cancer, and the fourth parotid gland malignancy. Their clinical and neuroimaging course is described, following chemotherapy for the malignant disease. None of the patients have suffered further MS outbreaks. The four women have improved and remain clinically stable after neoplastic treatment. Magnetic resonance imaging showed persistence of the same lesion burden in three patients, and reduction in the other. At the present one patient receives weekly intramuscular interferon-beta 1a, whereas the other did not received any treatment. CONCLUSIONS: Probably there are no specific cytostatics for MS. Immunosuppressive therapy could be a therapeutic option among patients with an aggressive clinical course.

Evolución clínica de pacientes con esclerosis múltiple tratados con citostáticos por cáncer / Clinical course of multiple sclerosis in patients treated with immunosuppressive drugs for cancer

Introducción. Aunque los citostáticos en dosis bajas se usan en la esclerosis múltiple (EM) por su efecto inmunosupresor, sus efectos secundarios a largo plazo limitan su utilización. En cambio, su uso es necesario en aquellos pacientes con EM que desarrollan un cáncer, pudiendo mejorar no sólo el cáncer, sino también su EM. Casos clínicos. Presentamos cuatro mujeres con EM clínicamente definida sin tratamiento previo que recibieron diversos citostáticos (cisplatino, 5-fluorouracilo, leucovorín, ciclofosfamida, adriamicina, tamoxifeno y anastrozol) tras haber desarrollado una neoplasia: cáncer de mama (2), cáncer de colon (1) y cáncer de parótida (1). Describimos la evolución clínica y de la neuroimagen tras haber sido tratadasdel cáncer con tratamiento inmunosupresor. Ninguna de las pacientes ha vuelto a presentar brotes de su EM. Las cuatro mejoraron y continúan clínicamente estables después de haber sido tratadas de su neoplasia. Una paciente está en tratamiento con interferón-beta 1a intramuscular semanal en el momento actual, mientras que las otras tres no reciben tratamiento alguno.En los estudios de resonancia magnética no se evidenciaron cambios significativos después del tratamiento en tres pacientes, mientras que en la otra se apreció una disminución en el número de lesiones (FLAIR) y carga lesional (T2). Conclusiones.Podrían no existir citostáticos específicos en el tratamiento de la EM. Aunque no son un tratamiento de primera elección, el tratamiento con citostáticos podría ser una opción terapéutica en los enfermos fundamentalmente con una evolución clínica más agresiva

Introduction. The risk of side effects secondary to global and non-specific immune suppression has limited the systematic application of immunosuppressive therapy in multiple sclerosis (MS). However, when a patient with MS develops a cancer, cytostatic drugs as treatment for the neoplastic process may induce improvement not only of the cancer but also of MS. Case reports. We present a series of four women with clinically defined MS and subjected to cytostatic therapy (cisplatin,5-fluorouracil, leukovorin, adriamycin, tamoxifen and anastrozole) after the development of cancer: two presented breast cancer, one colon cancer, and the fourth parotid gland malignancy. Their clinical and neuroimaging course is described, following chemotherapy for the malignant disease. None of the patients have suffered further MS outbreaks. The four womenhave improved and remain clinically stable after neoplastic treatment. Magnetic resonance imaging showed persistence of the same lesion burden in three patients, and reduction in the other. At the present one patient receives weekly intramuscular interferon-beta 1a, whereas the other did not received any treatment. Conclusions. Probably there are no specific cytostaticsfor MS. Immunosuppressive therapy could be a therapeutic option among patients with an aggressive clinical course

Clinical picture of a patient with a novel PSEN1 mutation (L424V).

Young onset dementia raises concern about familial and non degenerative dementias. We describe a patient with early dementia. At the age of 26, a woman developed symptoms of anorexia nervosa, at 30 a memory and attention deficit, and at 34 abnormal behavior with impulsivity, aggression, and dysexecutive disorder. At 36 she showed aphasia, stereotyped behavior, hyperreflexia, grasping reflex, urinary incontinence, myoclonus, and seizures. Blood and cerebrospinal fluid were normal. Brain computed tomography and single photon emission computed tomography showed diffuse cortico-subcortical atrophy and frontotemporoparietal hypoperfusion. A Leu424Val mutation was present in PSEN1 gene. PSEN1 mutations can produce Alzheimer's disease, frontotemporal dementia, and dementia with Lewy bodies phenotypes, or a combination of them. It has been proposed that the mutation type and location may influence the molecular pathogenesis and thus PSEN1 would represent a molecular connexion between these entities. This case shows a novel PSEN1 mutation with outstanding amnesic and frontal symptoms.

Prevalence of multiple sclerosis in Santiago de Compostela (Galicia, Spain).

INTRODUCTION: Multiple sclerosis (MS) is the most common central nervous system disease in young adults, and one of the leading causes of disability in this age group. OBJECTIVES: To establish the prevalence and incidence of MS in Santiago de Compostela (Spain). Material and methods We performed a prospective, case-finding study on > 90,188 inhabitants, from 1 January 1998 to 15 September 2003, selected as day of prevalence. Sources of information were the University Clinical Hospital of Santiago, the hospitals and private clinics of the city, rehabilitators, pediatricians, ophthalmologists, psychiatrists, neurosurgeons, specialists in internal medicine, primary care physicians, association of patients of MS, social services and nongovernmental and religious associations. Informative days were programmed for the susceptible population, and campaigns were carried out in the media. All patients with Poser diagnostic criteria for MS, on the census of Santiago de Compostela as of 15 September 2003, were included in the study. RESULTS: On the day of prevalence, 71 patients with MS were registered on the census of Santiago de Compostela, therefore, the prevalence was 79 cases per 100,000 inhabitants. The incidence of the disease in the study period was 5.3 cases per 100,000 inhabitants and year. CONCLUSIONS: The prevalence and incidence rates of MS found in Santiago de Compostela are the highest reported, to date, in Spain.

[Factors determining the duration of temporary disability and return to work in a health district of Galicia]. / Determinantes de la duración de la incapacidad temporal y la vuelta al trabajo en un área sanitaria de Galicia.

OBJECTIVE: To determine the factors associated with the incidence and duration of temporary work incapacity (TWI) in a health district. DESIGN: Descriptive and retrospective study. SETTING: South health district of the province of Lugo, Spain. PARTICIPANTS: A random sample of 1513 cases was selected among the total of episodes of TWI, during 3 years period. MAIN MEASURES: The main factors analyzed are, on the one hand, the socio-demographic characteristics of the patient, his or her social security (SS) scheme, diagnosis that justifies the TWD, and the prescription date; and, on the other hand, the age, sex, specialised training, time in the post and years in practice of the physician who prescribes the TWI. The comparison of the means was carried out using variance analysis and the Kruskal-Wallis test. The relative effect of each variable on the probability of returning to the work was estimated through Cox regression models. RESULTS: The mean duration of the episodes of TWI was of 74+/-103 days. The most frequent diagnoses were those of the bones-muscles and joints (BMAJ), injuries and poisonings (IAP), and respiratory diseases (RD). The probability of returning to work is reduced with the increase of the age, with agrarian and autonomous SS affiliates, with diagnoses of mental disease or diagnoses of the circulatory system, and in cases prescribed by older doctors or less time in the post. CONCLUSIONS: The mean duration of the episodes of TWD is higher than that of other Spanish studies. The most influential factors in the return to work are the age of the patient, the SS scheme and the diagnosed illness.

Determinantes de la duración de la incapacidad temporal y la vuelta al trabajo en un área sanitaria de Galicia / Factors determining the duration of temporary disability and return to work in a health district of Galicia

Objetivo. Determinar los factores asociados con la incidencia y la duración de la incapacidad temporal (IT) en un área sanitaria. Diseño. Descriptivo, retrospectivo. Emplazamiento. Área Sanitaria Sur de la provincia de Lugo. Participantes. Una muestra de 1.513 episodios de IT seleccionada aleatoriamente entre el total de éstos, durante un período de 3 años. Mediciones principales. Se analizaron las características sociodemográficas del paciente, el régimen de la seguridad social (SS), el diagnóstico que justifica la IT y la fecha de la prescripción; del médico prescriptor se analizaron la edad, el sexo, la formación especializada, la antigüedad en la plaza y los años de ejercicio. La comparación de medias se realizó mediante el análisis de la varianza y el test de Kruskal-Wallis. El efecto relativo de cada variable sobre la probabilidad de volver al trabajo se estimó mediante modelos de regresión de Cox. Resultados. La duración media de los episodios de IT fue de 74 ± 103 días. Los diagnósticos más frecuentes fueron los del sistema osteomioarticular (SOMA), las lesiones y envenenamientos (LYE) y las enfermedades respiratorias (NML). Se reduce la probabilidad de volver al trabajo con el incremento de la edad, en los regímenes de seguridad social autónomos y agrarios por cuenta propia, en los diagnósticos de enfermedades mentales y del aparato circulatorio, y cuando el médico prescriptor es de mayor edad o menos antiguo en la plaza. Conclusiones. La duración media de los episodios de IT es superior a la de otros estudios españoles. Los factores que más influyen en la reincorporación al trabajo son la edad del paciente, el régimen de la seguridad social y la enfermedad diagnosticada

Objective. To determine the factors associated with the incidence and duration of temporary work incapacity (TWI) in a health district. Design. Descriptive and retrospective study. Setting. South health district of the province of Lugo, Spain. Participants. A random sample of 1513 cases was selected among the total of episodes of TWI, during 3 years period. Main measures. The main factors analyzed are, on the one hand, the socio-demographic characteristics of the patient, his or her social security (SS) scheme, diagnosis that justifies the TWD, and the prescription date; and, on the other hand, the age, sex, specialised training, time in the post and years in practice of the physician who prescribes the TWI. The comparison of the means was carried out using variance analysis and the Kruskal-Wallis test. The relative effect of each variable on the probability of returning to the work was estimated through Cox regression models. Results. The mean duration of the episodes of TWI was of 74±103 days. The most frequent diagnoses were those of the bones-muscles and joints (BMAJ), injuries and poisonings (IAP), and respiratory diseases (RD). The probability of returning to work is reduced with the increase of the age, with agrarian and autonomous SS affiliates, with diagnoses of mental disease or diagnoses of the circulatory system, and in cases prescribed by older doctors or less time in the post. Conclusions. The mean duration of the episodes of TWD is higher than that of other Spanish studies. The most influential factors in the return to work are the age of the patient, the SS scheme and the diagnosed illness

GC-FID determination of cocaine and its metabolites in human bile and vitreous humor.

Gas chromatography was used in combination with flame ionization detection (GC-FID) to develop a method for determining cocaine and its two metabolites, benzoylecgonine (BEG) and ecgonine methyl ester (EME), in bile and vitreous humor. The method used a 12 m x 0.2 mm i.d. column of 0.33 microm film thickness packed with 5% phenylmethylsiloxane, and proadifen as a reference compound. Drug-free bile and vitreous humor samples were used to prepare solutions of the target compounds at concentrations over the range 0.1-4 microg ml(-1) that were subjected to solid-phase extraction through Bond Elut Certify columns and derivatized with 99:1 (v/v) N,O-bis-trimethylsilyltrifluoroacetamide (BSTFA)/trimethylchlorosilane (TMCS). Calibration graphs were highly linear, with correlation coefficients above 0.99 in all instances. Also, the precision of the method was found to be quite acceptable, with coefficients of variation less than 5% for bile and less than 7% for vitreous humor. The average extraction yields ranged from 73.6% to 91.2% for bile and from 71.5% to 92.2% for vitreous humor. The proposed method was used to analyse 26 samples of bile and as many of vitreous humor from individuals fatally poisoned by cocaine, whether alone or in combination with other drugs. The mean drug levels found were 0.75 and 1.54 microg ml(-1) for cocaine in bile and vitreous humor, respectively, 6.35 and 0.94 microg ml(-1) for BEG, and 2.18 and 0.61 microg ml(-1) for EME.