Bacterial succession and co-occurrence patterns of an enriched marine microbial community during light crude oil degradation in a batch reactor.

AIMS: The aim of this study was to investigate the dynamic changes in the bacterial structure and potential interactions of an acclimatized marine microbial community during a light crude oil degradation experiment. METHODS AND RESULTS: The bacterial community effectively removed 76·49% of total petroleum hydrocarbons after 30 days, as evidenced by GC-FID and GC-MS analyses. Short-chain alkanes and specific aromatic compounds were completely degraded within the first 6 days. High-throughput sequencing of 16S rRNA gene indicated that the starting bacterial community was mainly composed by Marinobacter and more than 30 non-dominant genera. Bacterial succession was dependent on the hydrocarbon uptake with Alcanivorax becoming dominant during the highest degradation period. Sparse correlations for compositional data algorithm revealed one operational taxonomic unit (OTU) of Muricauda and an assembly of six OTUs of Alcanivorax dieselolei and Alcanivorax hongdengensis as critical keystone components for the consortium network maintenance and stability. CONCLUSIONS: This work exhibits a stabilized marine bacterial consortium with the capability to efficiently degrade light crude oil in 6 days, under laboratory conditions. Successional and interaction patterns were observed in response to hydrocarbon consumption, highlighting potential interactions between Alcanivorax and keystone non-dominant OTUs over time. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results contribute to the understanding of interactions and potential roles of specific members of hydrocarbonoclastic marine bacterial communities, which will be useful for further bioaugmentation studies concerning the associations between indigenous and introduced micro-organisms.

Intermittent binge-like ethanol exposure during adolescence attenuates the febrile response by reducing brown adipose tissue thermogenesis in rats.

Ethanol (EtOH) consumption is a primary health risk worldwide, which generally starts during adolescence in a binge pattern (i.e., the episodic consumption of high amounts). Binge EtOH consumption can lead to modifications of the innate and adaptive immune responses, including fever. The present study evaluated the febrile response that was induced by lipopolysaccharide (LPS) and prostaglandins E2 (PGE2) and the mechanisms of thermoregulation in adolescent rats that were exposed to EtOH in a binge-like pattern. Male Wistar rats were treated with an intraperitoneal (i.p.) injection of EtOH or saline on postnatal days (PND) 25, 26, 29, 30, 33, 34, 37, and 38. On PND 51, they received a pyrogenic challenge with LPS (i.p.) or PGE2 (intracerebroventricular) to induce a febrile response. Interscapular brown adipose tissue (BAT) mass and uncoupling protein (UCP) activity in isolated mitochondria were evaluated on PND 51. The rats were then subjected to cold challenges to analyze adaptive thermogenesis. Intermittent EtOH exposure during adolescence impaired the LPS- and PGE2-induced febrile response 12 days after the end of EtOH exposure. Ethanol exposure decreased interscapular BAT mass, oxygen consumption, and UCP activity in isolated mitochondria, resulting in an impairment in thermogenesis at 5 °C. No morphological changes in BAT were observed. These findings indicate that binge-like EtOH exposure during adolescence impairs thermoregulation by reducing BAT mass and function. This reduction may last for a prolonged period of time after the cessation of EtOH exposure and may affect both cold defenses and the febrile response during the development of infectious diseases.

Cold stress on Araucaria angustifolia embryogenic cells results in oxidative stress and induces adaptation: implications for conservation and propagation.

Araucaria angustifolia (Bert.) O. Kuntze is a species critically endangered of extinction and its development and propagation is strongly affected by abiotic stress. We have previously shown the activation of uncoupling protein in A. angustifolia embryogenic stem cells subjected to cold stress. Now, we have furthered those studies by exposing these cells to cold stress (4 ± 1 °C for either 24 or 48 h) and evaluating parameters associated with oxidative stress and alterations in the cellular and mitochondrial responses. Cold stress affect the H2O2 levels and lipid peroxidation increased after both stress condition, an effect associated with the decrease in the activities of peroxidases, catalase and ascorbate/dehydroascorbate ratio. On the other hand, the activities of ascorbate peroxidase, monodehydroascorbate and dehydroascorbate reductases increased as an indication of adaptation. Another important impact of cold stress conditions was the decrease of external alternative NAD(P)H dehydrogenases activity and the increase of mitochondrial mass. These results show that cold stress induces oxidative stress in A. angustifolia embryogenic cells, which results in activation of the glutathione-ascorbate cycle as a compensation for the decrease in the activities of catalase, peroxidases, and external NAD(P)H dehydrogenases. Our results contribute to the understanding of the pathways that gymnosperms employ to overcome oxidative stress, which must be explored in order to improve the methods of conservation and propagation of A. angustifolia.

Plasma kinetics and biodistribution of a lipid emulsion resembling low-density lipoprotein in patients with acute leukemia.

Low-density lipoprotein (LDL) could be used as a carrier of chemotherapeutic agents to neoplastic cells that overexpress LDL receptors (rLDL), but LDL is difficult to obtain and handle. Recently, it was observed that a protein-free emulsion resembling the lipid portion of LDL (LDE) behave like native LDL when injected into the bloodstream. In this study, the evidence that LDE is taken up by rLDL was expanded by comparing LDL and LDE plasma decay curves in rabbits and by competition experiments with lymphocytes. To verify whether LDE could be removed from the plasma by neoplastic cells with increased rLDL, LDE labeled with 14Ccholesteryl ester was injected into 14 patients with acute myeloid leukemia (AML) and into 7 with acute lymphocytic leukemia (ALL). In AML rLDL expression is increased but in ALL it is normal. LDE plasma fractional clearance rate (FCR, in h-1) was calculated from the remaining radioactivity measured in plasma samples collected during 24 h following injection. LDE FCR was 3-fold greater in AML than in ALL patients 0.192 +/- 0.210 (SD) and 0.066 +/- 0.033 h-1, respectively, P < 0.035. When LDE injection was repeated in 9 AML patients in hematological remission, LDE FCR diminished 66% compared to the pretreatment values (from 0.192 +/- 0.210 to 0.065 +/- 0.038 h-1, P < 0.02), so that it could be estimated that nearly 66% of the emulsion was taken up by AML cells and only 34% by the normal tissues. As expected, LDE FCR was unchanged in 4 patients with ALL in hematological remission (0.069 +/- 0.044 h-1). Gamma camera images obtained 6 h after the injection of 99mTc-label LDE into one patient with ALL showed biodistribution similar to that of LDL. In one AML patient LDE was comparatively more concentrated over the areas corresponding to the bone marrow infiltrated by AML cells. Our results indicate that LDE FCR is increased in a disease known to contain malignant cells that overexpress rLDL, suggesting that LDE is taken up by malignant cells with increased rLDL.

Effect of MI-D, a new mesoionic compound, on energy-linked functions of rat liver mitochondria.

MI-D (4-phenyl-5-(4-nitro-cinnamoyl)-1,3,4-thiadiazolium-2-phenylami ne chloride), a new mesoionic compound, depressed the phosphorylation efficiency of liver mitochondria as deduced from an accentuated decrease of the respiratory control coefficient and ADP/O ratio. Analysis of segments of the respiratory chain suggested that the MI-D inhibition site is further on than complex I and between complexes II and III. The transmembrane electrical potential (delta psi) was collapsed dependent on MI-D concentration. ATPase activity was dramatically increased by MI-D in intact mitochondria, but inhibited in carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP)-uncoupled mitochondria. These results suggest that MI-D acts as an uncoupler agent, a property closely related to its structural characteristics.

Cardiovascular disease risk factors and diet of Fulani pastoralists of northern Nigeria.

BACKGROUND: The Fulani of northern Nigeria are seminomadic pastoralists who consume a diet rich in saturated fats, do not use tobacco, are lean, and have an active lifestyle. Little is known about their serum lipid profiles and corresponding risk of cardiovascular disease. OBJECTIVE: We measured serum lipid, homocysteine, folate, and vitamin B-12 concentrations in Fulani men and women and assessed the nutrient content of their diet. DESIGN: Blood samples from 42 men (18-64 y old) and 79 women (15-77 y old) living in the Jos Plateau of Nigeria were analyzed for cholesterol (total, HDL, and LDL), triacylglycerol, homocysteine, folate, and vitamin B-12 serum concentrations. Body composition was determined by bioelectrical impedance analysis. Dietary information was obtained with use of a 7-d dietary recall and a food-frequency questionnaire. Results were compared with US referent ranges. RESULTS: The mean energy content of the Fulani diet was relatively low (men, 6980 kJ; women, 6213 kJ) and the mean protein content was high (men, 20% of energy; women, 16% of energy). Nearly one-half of energy was provided by fat, and one-half of that was derived from saturated fatty acids. The diet provided marginal to adequate amounts of vitamins B-12, B-6, and C but only one-third of the US recommended dietary allowance for folate. The mean total cholesterol, HDL-cholesterol, and triacylglycerol concentrations of Fulani adults were within the referent ranges; the mean LDL-cholesterol concentration of Fulani adults below the range; and the mean serum homocysteine concentration of Fulani men above the range. Homocysteine and folate concentrations were inversely correlated for both men and women. CONCLUSIONS: Despite a diet high in saturated fat, Fulani adults have a lipid profile indicative of a low risk of cardiovascular disease. This finding is likely due to their high activity level and their low total energy intake.

Sister-chromatid exchange frequencies induced by metrifonate in mammalian in vivo and in vitro systems.

Metrifonate is a drug widely used to cure several helminthiases in man including cysticercosis. In this study we determine its capacity to induce SCEs in vivo and in vitro using the fluorescence-plus-Giemsa technique. In the human lymphocyte system we tested 10, 20, 30, 40, 50 and 60 micrograms/ml added at 24 h of culture with a duration of 72 h. The results showed a SCE inducing effect with the exception of the first tested dose (10 micrograms/ml), but without a dose-response expression. A cell cycle progression delay was seen with the three highest dosages. In the mouse bone marrow we tested 30, 60 and 120 mg/kg in a 24-h experiment, and the results indicated an SCE increase only with the highest dose. No modification of the cell proliferation kinetics was observed.

Leukocytes Detection, Classification and Counting in Smears of Peripheral Blood / Detección, Clasificación y Conteo de Leucocitos en Frotis de Sangre Periférica

Using the k-NN classifier in combination with the first Minkowski metric, in addition to techniques of digital image processing, we developed a computational system platform-independent, which is able to identify, to classify and to count five normal types of leukocytes: neutrophils, eosinophils, basophils, monocytes and lymphocytes. It is important to emphasize that this work does not attempt to diferentiate between smears of leukocytes coming from healthy and sick people; this is because most diseases produce a change in the differential count of leukocytes rather than in theirs forms. In the other side, the system could be used in emerging areas such as the topographic hematology and the chronobiology.

Mediante un clasificador k-NN en combinación con la primera métrica de Minkowski y técnicas de procesamiento digital de imágenes, se desarrolló un sistema computacional independiente de la plataforma, capaz de identificar, clasificar y contar cinco formas normales de leucocitos: neutrófilos, eosinófilos, basófilos, monocitos y linfocitos. Es importante enfatizar que este trabajo no intenta diferenciar entre muestras de leucocitos provenientes de gente sana y enferma, debido a que la mayoría de las enfermedades se detectan principalmente por un cambio en el conteo diferencial de leucocitos más que por cambios en su forma. Finalmente, el contador de leucocitos puede ser usado en áreas emergentes como la hematología topográfica y la cronobiología.

Cell protection induced by beta-sitosterol: inhibition of genotoxic damage, stimulation of lymphocyte production, and determination of its antioxidant capacity.

Beta-sitosterol (BS) is a compound that has shown various activities potentially useful for human health. In the present study, we determined its antigenotoxic capacity and lymphocyte induction potential in mouse as well as its capacity to trap free radicals in vitro. BS, in doses from 200 to 1,000 mg/kg, was able to significantly reduce the frequency of sister chromatid exchanges induced by 10 mg/kg of doxorubicin (DX) in bone marrow cells. The same range of BS doses also gave rise to a strong reduction in the rate of micronucleated, polychromatic erythrocytes induced by DX. In addition, we determined an increase in the production of lymphocytes in mice administered with BS. By means of the DPPH assay, the compound was shown to trap free radicals in a concentration dependent manner as high as 78.12% using 250 mug/ml. Our research established three relevant biological activities of BS which show its potential as a chemopreventive agent.

Genotoxic and cytotoxic studies of beta-sitosterol and pteropodine in mouse.

Beta-sitosterol (BS) and pteropodine (PT) are constituents of various plants with pharmacological activities potentially useful to man. The chemicals themselves possess biomedical properties related to the modulation of the immune and the nervous systems, as well as to the inflammatory process. Therefore, safety evaluation of the compounds is necessary in regard to their probable beneficial use in human health. The present study evaluates their genotoxic and cytotoxic potential by determining the capacity of the compounds to induce sister chromatid exchanges (SCE), or to alter cellular proliferation kinetics (CPK) and the mitotic index (MI) in mouse bone marrow cells. Besides, it also determines their capacity to increase the rate of micronucleated polychromatic erythrocytes (MNPE) in peripheral mouse blood, and the relationship polychromatic erythrocytes/normochromatic erythrocytes (PE/NE) as an index of cytotoxicity. For the first assay, four doses of each compound were tested: 200, 400, 600, and 1000 mg/kg in case of BS, and 100, 200, 300, and 600 mg/kg for PT. The results in regard to both agents showed no SCE increase induced by any of the tested doses, as well as no alteration in the CPK, or in the MI. With respect to the second assay, the results obtained with the two agents were also negative for both the MNPE and the PE/NE index along the daily evaluation made for four days. In the present study, the highest tested dose corresponded to 80% of the LD(50) obtained for BS and to 78% in the case of PT. The results obtained establish that the studied agents have neither genotoxic nor cytotoxic effect on the model used, and therefore they encourage studies on their pharmacological properties.

Escaso conocimiento del paciente con prótesis valvular cardiaca / Scant knowledge of heart valvular protheses patients recarding their disease

Dentro de las cirugías cardiacas la valvular es una de las más frecuente en el adulto; para obtener buenos resultados se requiere de medidas especiales y educación del paciente. Por esta razón se diseño un estudio prospectivo y observacional que consta de un cuestionario de 11 preguntas sobre el conocimiento de la enfermedad vascular, que se aplicó a 126 pacientes con prótesis vascular cardiaca. En promedio respondieron cinco preguntas, ninguno contesto las 11. Este estudio demuestra el poco conocimiento de los pacientes acerca de su enfermedad, el cual los médicos también deben intervenir