Identification of AREB/ABF Gene Family Involved in the Response of ABA under Salt and Drought Stresses in Jute (Corchorus olitorius L.).

The abscisic acid (ABA)-responsive element binding protein/ABRE-binding factor (AREB/ABF) subfamily members are essential to ABA signaling pathways and plant adaptation to various environmental stresses. Nevertheless, there are no reports on AREB/ABF in jute (Corchorus L.). Here, eight AREB/ABF genes were identified in the C. olitorius genome and classified into four groups (A-D) based on their phylogenetic relationships. A cis-elements analysis showed that CoABFs were widely involved in hormone response elements, followed by light and stress responses. Furthermore, the ABRE response element was involved in four CoABFs, playing an essential role in the ABA reaction. A genetic evolutionary analysis indicated that clear purification selection affects jute CoABFs and demonstrated that the divergence time was more ancient in cotton than in cacao. A quantitative real-time PCR revealed that the expression levels of CoABFs were upregulated and downregulated under ABA treatment, indicating that CoABF3 and CoABF7 are positively correlated with ABA concentration. Moreover, CoABF3 and CoABF7 were significantly upregulated in response to salt and drought stress, especially with the application of exogenous ABA, which showed higher intensities. These findings provide a complete analysis of the jute AREB/ABF gene family, which could be valuable for creating novel jute germplasms with a high resistance to abiotic stresses.

Stability of Cocaine Compounds in Biological Fluids During Post-Analytical Sample Storage.

The objective of this study is to evaluate in vitro stability of cocaine compounds, cocaine (COC), benzoylecgonine (BE), ecgonine methyl ester (EME) and benzoylecgonine ethyl ester (EBE), in blood and urine, during post-analysis custody. Stability was evaluated by measuring percent recovery. Parameters evaluated were time of custody (1 year), storage temperature (-20°C and 4°C), influence of preservative (only for blood samples) and pH (only for urine samples). The impact of the temperature is very important in blood samples. At -20°C all compounds demonstrated to be stable, with recoveries higher than 80% after 1 year. In contrast, degradation was observed in the concentration for all four compounds when the samples were maintained at 4°C. In these same conditions, the influence of the preservative was also noticeable and a higher stability was found in samples preserved with NaF. COC and EBE had similar profiles, and both compounds disappeared after 30 days in samples without NaF and after 150 days in samples with NaF added. EME disappeared after 185 days and after 215 days in samples without and with preservative, respectively. BE recoveries, after 365 days of storage, were 68.5% (in samples with NaF) and 3.7% (in samples without NaF). In urine samples, the four compounds were stable in all the studied conditions except when samples were at pH 8 and stored at 4°C where the compounds disappeared (COC and EBE after 75 days of storage and EME after 15 days). The exception was BE, with a recovery of 23% after 1 year of storage. Of the temperatures evaluated, -20°C seems to be optimal for storage to maintain the stability of cocaine and metabolites in biological samples. This can be further enhanced by maintaining a pH of 4 in urine samples and adding a NaF preservative to blood.

Genetic variant affecting the myosin light chain 2 related to familial hypertrophic cardiomyopathy.

Familial hypertrophic cardiomyopathy (FHCM) is a genetic disease characterized by left ventricle (LV) or interventricular septum hypertrophy. FHCM is a common heart disease (affecting 1 out of 500 individuals) associated with genetic variants in genes related to the sarcomere, including the MYL2 (myosin light chain 2) gene that is affected in 1 to 3% of the cases. As described in this report, the genetic mutation p.Gly87Ala, rs 397516399 in the MYL2 gene is likely pathogenic. Reported here is the case of a 37-year-old Colombian man with asymmetric septal hypertrophic cardiomyopathy and ventricular tachycardia. The man had progressive symptomatology, a family history of FHCM with a dominant inheritance pattern, a mother and 2 brothers with FHCM, and 2 brothers who died suddenly before the age of 35. A molecular panel of 17 genes for hypertrophic cardiomyopathy identified a heterozygous variant, p.Gly87Ala, of the MYL2 gene. This variant can be found in Ensembl, dbSNP, and ClinVar, where it has conflicting interpretations: it either has an uncertain significance or it is likely pathogenic. This is the first report of a Colombian case of FHCM secondary to a mutation in the MYL2 gene, highlighting the importance of molecular diagnosis, genetic counseling, and bioinformatic analysis in these patients.

Nivel de conocimiento y frecuencia de autoexamen de mama en alumnos de los primeros años de la carrera de Medicina / Level of knowledge and frequency of breast self-exam in medical students

Determinar la frecuencia y el conocimiento de la práctica del autoexamen de mama en alumnos de los tres primeros años de la carrera de Medicina en una universidad privada de Lima, Perú. Material y métodos: Estudio transversal descriptivo en 281 estudiantes que consistió en una encuesta de once preguntas diseñadas para evaluar la frecuencia de la práctica y el conocimiento del autoexamen de mama. Resultados: El 89,7% de los participantes había oído del autoexamen de mama; el 21,0% lo practicaba y el 41,0% de ellos lo hacía mensualmente. En términos de conocimiento, la media fue 6,8 ± 1,8 puntos; solo el 1,0% alcanzó el puntaje máximo de once puntos. Conclusiones: La práctica del autoexamen es baja; pero parece haber un conocimiento aceptable. Es importante enfocar el esfuerzo de las intervenciones en promover la práctica de este método de tamizaje temprano...

To determine the frequency, level of knowledge and practice of breast auto-exam among first three-year medical students of a private University in Lima, Peru. Methods: Cross-sectional study performed in 281 students who answered a survey aimed at evaluating the objectives of the study. Results: 89.7% of participants have heard of breast self-exam; 21% practiced it, and 41% performed it monthly. Mean knowledge score was 6.8 ± 1.8 points; only 1% of participants attained the maximum score of 11 points. Conclusions: The frequency of breast self-exam is low despite of an acceptable level of knowledge. Efforts should be implemented to promote breast self-exam...

Specific characteristics of sudden death in a mediterranean Spanish population.

Most of the data reported on sudden cardiac death has been from studies of Anglo-Saxon patients. We conducted a study to ascertain the relation between sudden death (SD) and some epidemiologic, clinical, and biochemical parameters and to assess the coronary histopathologic aspects of subjects in a Spanish population who had died suddenly. A total of 204 subjects (86% men), aged 12 to 80 years (mean 54 ± 15), who had died from out-of-hospital natural SD were evaluated. Only 15% of subjects had been previously diagnosed with heart disease. Pathologic evidence of underlying cardiovascular disease was found in 90% of cases, with coronary heart disease (CHD) the most frequent (58%). The CHD was acute coronary thrombosis in 41% and a stable plaque with luminal narrowing of ≥75% in 59%. An old myocardial infarction was found in 31% of the SD victims. Cardiac hypertrophy was found in 48%, with no relation between the presence of cardiac hypertrophy and CHD. Patients with stable plaques had a greater heart weight than did those with acute coronary thrombosis (p = 0.02). Male gender, older age, smoking, and low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio of ≥3 were associated with CHD. A greater percentage of patients with an eroded and/or ruptured plaque than patients with a stable plaque were smokers. Only smoking and a low-density lipoprotein/high-density lipoprotein cholesterol ratio of ≥3 were associated with an eroded and/or ruptured plaque. In conclusion, compared with the findings from studies of Anglo-Saxon patients, a lower incidence of CHD and acute coronary thrombosis and a greater incidence of cardiac hypertrophy were found in SD victims of a Mediterranean Spanish population.

Cocaine-related sudden death: a prospective investigation in south-west Spain.

AIMS: With an estimated 12 million consumers in Europe, cocaine (COC) is the illicit drug leading to the most emergency department visits. The aim of this study was to examine a consecutive series of sudden deaths (SDs) to focus on the prevalence, the toxicological characteristics, and the causes of death in COC-related fatalities. METHODS AND RESULTS: Prospective case-control study of forensic autopsies was carried out in the time interval November 2003 to June 2006 at the Institute of Legal Medicine, Seville, south-west Spain, with a reference population of 1 875 462 inhabitants. Toxicology included blood ethanol analysis and blood and urine investigation for drugs of abuse and medical drugs. Autopsy was performed according to the European standardized protocol. Ten age- and sex-matched patients who died of violent causes with no antecedents of COC consumption and negative toxicology served as controls. During the study period, 2477 forensic autopsies were performed, including 1114 natural deaths. Among the latter, 668 fulfilled the criteria of SD and 21 (all males, mean age 34.6 +/- 7.3 years) resulted to be COC-related (3.1%). Cocaine was detected in 67.1% of the blood (median 0.17 mg/L, interquartile range 0.08-0.42) and in 83.0% of the urine samples (median 1.15 mg/L, interquartile range 0.37-17.34). A concomitant use of ethanol was found in 76.0% and cigarette smoking in 81.0%. Causes of SD were cardiovascular in 62.0%, cerebrovascular in 14.0%, excited delirium in 14.0%, respiratory and metabolic in 5.0% each. Left ventricular hypertrophy was observed in 57.0%, small vessels disease in 42.9%, severe atherosclerotic coronary artery disease in 28.6%, and coronary thrombosis in 14.3%. CONCLUSION: Systematic toxicology investigation indicates that 3.1% of SDs are COC-related and are mainly due to cardio-cerebrovascular causes. Left ventricular hypertrophy, small vessel disease, and premature coronary artery atherosclerosis, with or without lumen thrombosis, are frequent findings that may account for myocardial ischaemia at risk of cardiac arrest in COC addicts.

Pulmonary embolism and sudden-unexpected death: prospective study on 2477 forensic autopsies performed at the Institute of Legal Medicine in Seville.

Pulmonary embolism (PE) and deep venous thrombosis (DVT) are clinical manifestations of the same entity, venous thromboembolic disease (VTD). In approximately 25% of patients, the first manifestation of PE is sudden-unexpected death. We carried out a prospective study at the forensic pathology service of the Institute of Legal Medicine in Seville with the aim to know the incidence of PE as well as to describe the epidemiological, pathological and clinical characteristics of these deaths and associated risk factors. In the study period (32 months) 32 cases of PE were registered from a total of 2447 completed autopsies. Three cases were considered accidental deaths and the remaining 29 cases were sudden natural deaths, which represents 1.3% of the total autopsies, 2.6% of natural deaths and 4.3% of sudden deaths. Nineteen cases (59%) were men (mean age 50.3+/-13.8, range 22-74 years) and 13 cases (41%) were women (mean age 50.3+/-13.8, range 18-87 years). In 78% of cases death occurred at home or during transfer to a health care centre, mainly during the fall or winter (69%) and between 8a.m. and 4p.m. (47%). Pulmonary infarction was associated only in two cases (6%). Nine cases (28%) had been immobilized but only three (9%) received anticoagulant therapy. Surgical interventions had occurred in seven cases (22%). A history of psychiatric pathology was found in 31%. Overweight or obesity was found in 75%. The most frequent symptoms prior to death were dyspnea (31%) and chest pain (19%), and 19% of patients were examined in an Emergencies Department for symptoms compatible with deep vein thrombosis and/or PE, but this diagnosis was not suspected in any case. PE frequently makes its first appearance as sudden death. In addition to the classic risk factors, this study highlights that 75% of the cases were overweight/obese as well as 31% having had a history of psychiatric disorders and treatment as to support that this association should be considered as a risk factor. PE continues to be under diagnosed in Emergencies Department patients, which hinders the application of adequate therapeutic measures to prevent these deaths.

Commotio cordis as a result of a fight: report of a case considered to be imprudent homicide.

Commotio cordis is a clinic-pathological syndrome related to sudden death in young people involved in sports activities. It has been described, mainly, in athletes without previous cardiac anomalies who received a minor blow to the chest which produces ventricular fibrillation and cardiac arrest in the absence of structural damage to the ribs, sternum, or heart. There are few reported cases of commotio cordis associated with violent, non-sports related actions, which are commonly considered to be imprudent homicides. We present the case of a 20-year-old man, who was kicked in the chest during a fight; he suddenly collapsed although advanced cardio-respiratory resuscitation started shortly. Autopsy showed no cardiac lesions concluding that death was due to commotio cordis (blunt trauma to the chest). Toxicological analysis determined the presence of 5.14 mg/L benzoylecgonine in blood. On the basis of medico-legal investigation, the official prosecution considered the death to be imprudent homicide and the aggressor was sentenced to 4 years in prison. We emphasize the importance of the knowledge of the death circumstances through the witnesses' testimony, prior to beginning the autopsy, to confirm this important medico-legal diagnosis. Arrhythmogenic effects of cocaine and its contribution in the production of these deaths are also exposed.

Toxicological assessment of indium nitrate on aquatic organisms and investigation of the effects on the PLHC-1 fish cell line.

Indium nitrate is mainly used as a semiconductor in batteries, for plating and other chemical and medical applications. There is a lack of available information about the adverse effects of indium compounds on aquatic organisms. Therefore, the toxic effects on systems from four trophic levels of the aquatic ecosystem were investigated. Firstly, the bacterium Vibrio fischeri, the alga Chlorella vulgaris and the cladoceran Daphnia magna were used in the toxicological evaluation of indium nitrate. The most sensitive model was V. fischeri, with a NOAEL of 0.02 and an EC(50) of 0.04 mM at 15 min. Although indium nitrate should be classified as harmful to aquatic organisms, it is not expected to represent acute risk to the aquatic biota. Secondly, PLHC-1 fish cell line was employed to investigate the effects and mechanisms of toxicity. Although protein content, neutral red uptake, methylthiazol metabolization, lysosomal function and acetylcholinesterase activity were reduced in cells, stimulations were observed for metallothionein levels and succinate dehydrogenase and glucose-6-phosphate dehydrogenase activities. No changes were observed in ethoxyresorufin-O-deethylase activity. To clarify the main events in PLHC-1 cell death induced by indium nitrate, nine modulators were applied. They were related to oxidative stress (alpha-tocopherol succinate, mannitol and sodium benzoate), disruption of calcium homeostasis (BAPTA-AM and EGTA), thiol protection (1,4-dithiotreitol), iron chelation (deferoxiamine) or regulation of glutathione levels (2-oxothiazolidine-4-carboxylic acid and malic acid diethyl ester). The main morphological alterations were hydropic degeneration and loss of cells. At least, in partly, toxicity seems to be mediated by oxidative stress, and particularly by NADPH-dependent lipid peroxidation.

Ecotoxicological effects of the antioxidant additive propyl gallate in five aquatic systems.

Propyl gallate is an antioxidant widely used in foods, cosmetics and pharmaceuticals. The occurrence and fate of additives in the aquatic environment is an emerging issue in environmental chemistry. To date, there is little available information about the adverse effects of propyl gallate on aquatic organisms. Therefore, the toxic effects were investigated, using five model systems from four trophic levels. The most sensitive system was the hepatoma fish cell line PLHC-1 according to total protein content, with an EC(50) of 10 microM and a NOAEL of 1 microM at 72 h, followed by the immobilization of Daphnia magna, the inhibition of bioluminescence of Vibrio fischeri, the salmonid fish cell line RTG-2 and the inhibition of the growth of Chlorella vulgaris. Although protein content, neutral red uptake, methylthiazol metabolization and acetylcholinesterase activity were reduced in PLHC-1 cells, stimulations were observed for lysosomal function, succinate dehydrogenase, glucose-6-phosphate dehydrogenase and ethoxyresorufin-O-deethylase activities. No changes were observed in metallothionein levels. The main morphological observations were the loss of cells and the induction of cell death mainly by necrosis but also by apoptosis. The protective and toxic effects of propyl gallate were evaluated. General antioxidants and calcium chelators did not modify the toxicity of propyl gallate, but an iron-dependent lipid peroxidation inhibitor gave 22% protection. The results also suggest that propyl gallate cytotoxicity is dependent on glutathione levels, which were modulated by malic acid diethyl ester and 2-oxothiazolidine-4-carboxylic acid. According to the results, propyl gallate should be classified as toxic to aquatic organisms.

Ecotoxicological evaluation of sodium fluoroacetate on aquatic organisms and investigation of the effects on two fish cell lines.

Sodium monofluoroacetate (compound 1080) is one of the most potent pesticides. It is also a metabolite of many other fluorinated compounds, including anticancer agents, narcotic analgesics, pesticides or industrial chemicals. Other sources of water contamination are the atmospheric degradation of hydrofluorocarbons and hydrochlorofluorocarbons. However, there is little information available about the adverse effects of sodium fluoroacetate in aquatic organisms. Firstly, the bacterium Vibrio fischeri (decomposer), the alga Chlorella vulgaris (1st producer) and the cladoceran Daphnia magna (1st consumer) were used for the ecotoxicological evaluation of SMFA. The most sensitive models were C. vulgaris and D. magna, with a NOAEL of 0.1 and an EC50 of 0.5 mM at 72 h, respectively. According to the results after the acute exposure and due to its high biodegradation rate and low bioaccumulation potential, sodium fluoroacetate is most unlikely to produce deleterious effects to aquatic organisms. Secondly, two fish cell lines were employed to investigate the effects and mechanisms of toxicity in tissues from 2nd consumers. The hepatoma fish cell line PLHC-1 was more sensitive to SMFA than the fibroblast-like fish cell line RTG-2, being the uptake of neutral red the most sensitive bioindicator. Lysosomal function, succinate dehydrogenase and acetylcholinesterase activities were inhibited, glucose-6-phosphate dehydrogenase activity was particularly stimulated, and metallothionein and ethoxyresorufin-O-deethylase levels were not modified. Intense hydropic degeneration, macrovesicular steatosis and death mainly by necrosis but also by apoptosis were observed. Moreover, sulphydryl groups and oxidative stress could be involved in PLHC-1 cell death induced by SMFA more than changes in calcium homeostasis.

Toxicological effects of the lipid regulator gemfibrozil in four aquatic systems.

Gemfibrozil is a lipid-regulating agent widely used in patients at risk of coronary disease. Pharmaceutical products, such as gemfibrozil, are found in municipal effluents and represent a major source of contamination. To date, there is little available information about the adverse effects of gemfibrozil in aquatic organisms. For this reason, the toxic effects were investigated using model systems from four trophic levels. The most sensitive system was the immobilization of Daphnia magna, with a non-observed adverse effect level of 30 microM and a mean effective concentration of 120 microM after 72 h, followed by the inhibition of bioluminescence of Vibrio fischeri, the hepatoma fish cell line PLHC-1 line and the inhibition of the growth of Chlorella vulgaris. Although protein content, neutral red uptake, methylthiazol metabolization and lysosomal function were reduced in PLHC-1 cells, stimulations were observed for lysosomal function, metallothionein levels and succinate dehydrogenase, glucose-6-phosphate dehydrogenase and acetylcholinesterase activities. No changes were observed in ethoxyresorufin-O-deethylase activity. The main morphological alterations were hydropic degeneration and loss of cells. Modulation studies on gemfibrozil toxicity were also carried out. General antioxidants and calcium chelators did not modify the toxicity of gemfibrozil, whereas a Fe(III) chelator, a membrane permeable sulphydryl-protecting compound and glutathione level modifying agents did change the toxicity. One of the possible mechanisms of gemfibrozil toxicity seems to be the binding to sulphydryl groups, including those of glutathione. According to the result, gemfibrozil should be classified as harmful to aquatic organisms. However, comparing the concentrations in water and the toxicity quantified in the assayed systems, gemfibrozil is not expected to represent acute risk to the aquatic biota.

Ecotoxicological assessment of bromobenzene using a test battery with five model systems.

Bromobenzene (BrB) is used as a solvent for crystallization and as an additive to motor oils and may be released into the environment through various waste streams. However, there is limited available information about the toxic hazard of BrB in the aquatic environment. Consequently, the ecotoxicological effects induced by BrB were investigated using five model systems with representants from four trophic levels. The battery included bioluminescence inhibition of the bacterium Vibrio fischeri, growth inhibition of the alga Chlorella vulgaris and immobilization of the cladoceran Daphnia magna. Total protein content, neutral red uptake and MTS metabolization were reduced, while lysosomal function, succinate dehydrogenase activity, G6PDH activity and leakage, metallothionein levels and EROD activity were stimulated in PLHC-1 and RTG-2 fish cell lines. The most sensitive bioindicator was the bioluminiscence of V. fischeri, with an EC(50) of 0.04mM BrB at 15min and a non-observed adverse effect level of 0.02 mM BrB. There is a large difference in sensitivity to BrB among the model systems probably due to the metabolic capacity of the different species. PLHC-1 cells were more sensitive to BrB than RTG-2 cells. The most prominent morphological effects observed were hydropic degeneration, loss of cells and of the perinuclear pattern of distribution of lysosomes. Therefore, BrB should be classified as toxic to aquatic organisms.

Ecotoxicological evaluation of the antimalarial drug chloroquine.

There is limited information available about the potential environmental effects of chloroquine (CQ), a widely used antimalarial agent and a promising inexpensive drug in the management of HIV disease. The acute effects of CQ were studied using four ecotoxicological model systems. The most sensitive bioindicator was the immobilization of the cladoceran Daphnia magna, with an EC50 of 12 microM CQ at 72 h and a non-observed adverse effect level of 2.5 microM CQ, followed very closely by the decrease of the uptake of neutral red and the reduction of the lysosomal function in the fish cell line PLHC-1 derived from the top minnow Poeciliopsis lucida, probably due to the selective accumulation of the drug into the lysosomes. There was significant cellular stress as indicated by the increases on metallothionein and glucose-6P dehydrogenase levels after 24 h of exposure and succinate dehydrogenase activity mainly after 48 h. No changes were observed for ethoxyresorufin-O-deethylase (EROD) activity. The least sensitive model was the inhibition of bioluminescence in the bacterium Vibrio fischeri. An increase of more than five-fold in the toxicity from 24 to 72 h of exposure was observed for the inhibition of the growth in the alga Chlorella vulgaris and the content of total protein and MTS tetrazolium salt metabolization in PLHC-1 cells. At the morphological level, the most evident alterations in PLHC-1 cultures were hydropic degeneration from 25 microM CQ after 24h of exposure and the presence of many cells with pyknotic nuclei, condensed cytoplasm and apoptosis with concentrations higher than 50 microM CQ after 48 h of exposure. In conclusion, CQ should be classified as harmful to aquatic organisms.

Ecotoxicological evaluation of the additive butylated hydroxyanisole using a battery with six model systems and eighteen endpoints.

The occurrence and fate of additives in the aquatic environment is an emerging issue in environmental chemistry. This paper describes the ecotoxicological effects of the commonly used additive butylated hydroxyanisole (BHA) using a test battery, comprising of several different organisms and in vitro test systems, representing a proportion of the different trophic levels. The most sensitive system to BHA was the inhibition of bioluminescence in Vibrio fischeri bacteria, which resulted in an acute low observed adverse effect concentration (LOAEC) of 0.28 microM. The next most sensitive system was the immobilization of the cladoceran Daphnia magna followed by: the inhibition of the growth of the unicellular alga Chlorella vulgaris; the endpoints evaluated in Vero (mammalian) cells (total protein content, LDH activity, neutral red uptake and MTT metabolization), mitotic index and root growth inhibition in the terrestrial plant Allium cepa, and finally, the endpoints used on the RTG-2 salmonid fish cell line (neutral red uptake, total protein content, MTS metabolization, lactate dehydrogenase leakage and activity, and glucose-6-phosphate dehydrogenase activity). Morphological alterations in RTG-2 cells were also assessed and these included loss of cells, induction of cellular pleomorphism, hydropic degeneration and induction of apoptosis at high concentrations. The results from this study also indicated that micronuclei were not induced in A.cepa exposed to BHA. The differences in sensitivity for the diverse systems that were used (EC50 ranged from 1.2 to >500 microM) suggest the importance for a test battery approach in the evaluation of the ecological consequences of chemicals. According to the results, the levels of BHA reported in industrial wastewater would elicit adverse effects in the environment. This, coupled with its potential to bioaccumulate, makes BHA a pollutant of concern not only for acute exposures, but also for the long-term.