Caracterización de un Consorcio Microbiano Metanogénico de una Mina de Carbón en la Cuenca de Bogotá / Characterization of a Methanogenic Microbial Consortium from a Coal Mine in Bogotá Basin

RESUMEN En el trabajo se estudió un consorcio microbiano metanogénico de una mina de carbón de la cuenca de Bogotá en Colombia. Se establecieron cultivos de enriquecimiento de carbón ex situ para el crecimiento y la producción de gas de novo. El gas biogénico producido por los cultivos se analizó mediante cromatografía de gases con detectores de ionización de llama y conductividad térmica. Los cultivos se utilizaron para aislar estirpes microbianas y para generar bibliotecas del gene 16S rARN empleando de cebadores de bacteria y de arquea. El análisis de cromatografía de gases mostró producción de metano a 37 oC, pero no a 60 oC, donde el CO2 fue el componente principal del gas biogénico. El análisis de la secuencia del gen 16S rARN de estirpes microbianos y de las bibliotecas de clones, estableció que el consorcio microbiano metanogénico estuvo formado por especies de bacterias de los géneros Bacillus y Gracilibacter más la arquea del género Methanothermobacter. El consorcio microbiano metanogénico identificado es potencialmente responsable de la generación de gas biogénico en la mina de carbón La Ciscuda. Los resultados sugirieron que los metanógenos de este consorcio producían metano por vía hidrogenotrófica o de reducción de CO2.

ABSTRACT The work studied the methanogenic microbial consortium in a coal mine from the Bogotá basin in Colombia. Ex situ coal-enrichment cultures were established for in vitro growth and de novo gas production. Biogenic gas produced by cultures was analyzed by gas chromatography using thermal conductivity and flame ionization detectors. Cultures were used to isolate microbial specimens and to generate 16S rRNA gene libraries employing bacterial and archaeal primer sets. The gas chromatographic analysis showed methane production at 37 oC, but not at 60 oC, where CO2 was the major component of the biogenic gas. 16S rRNA gene sequence analysis of microbial isolates and clone libraries established that the methanogenic microbial consortium was formed by bacteria species from Bacillus and Gracilibacter genera plus archaea from the Methanothermobacter genus. This meth-anogenic microbial consortium was potentially responsible for biogenic gas generation in La Ciscuda coal mine. The results suggested that these methanogens produced methane by hydrogenotrophic or CO2 reduction pathways.

Proteomic Analysis Reveals That an Extract of the Plant Lippia origanoides Suppresses Mitochondrial Metabolism in Triple-Negative Breast Cancer Cells.

Triple-negative breast cancer is an aggressive subtype of breast cancer with low 5-year survival rates, high 3-year recurrence rates, and no known therapeutic targets. Recent studies have indicated that triple-negative breast cancers possess an altered metabolic state with higher rates of glycolysis, mitochondrial oxidative phosphorylation, and increased generation and utilization of tricarboxylic acid cycle intermediates. Here, we utilized label-free quantitative proteomics to gain insight into the anticancer mechanisms of a methanolic extract from the Central American plant Lippia origanoides on MDA-MB-231 triple-negative breast cancer cells. The L. origanoides extract dysregulated mitochondrial oxidative phosphorylation by suppressing the expression of several subunits of Complex I of the electron transport chain, and inhibited cellular metabolism by down-regulating key tricarboxylic acid cycle enzymes and mitochondrial lipid and amino-acid metabolic pathways. Our study also revealed that treatment with the extract activated the stress response and pathways related to cell-cycle progression and DNA repair. Overall, our results reveal compelling new evidence that the extract from L. origanodes triggers rapid irreversible apoptosis in MDA-MB-231 cells by effectively 'starving' the cells of metabolites and ATP. We continue to study the specific bioactive components of the extract in the search for novel, highly effective mitochondrial inhibitors to selectively target triple-negative breast cancer.

Lippia origanoides extract induces cell cycle arrest and apoptosis and suppresses NF-κB signaling in triple-negative breast cancer cells.

Treatments targeting hormone receptors typically fail to provide a positive clinical outcome against triple-negative breast cancers (TNBC), which lack expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her2/neu). Towards identifying viable treatments for aggressive breast cancer, we have tested an extract of the tropical plant Lippia origanoides (LOE) on TNBC and normal cells lines to uncover its potential anticancer effects. Treatment with LOE reduced TNBC cell viability in a dose-dependent manner to a greater extent than in normal mammary epithelial MCF10A cells. In MDA-MB­231 cells, LOE was found to halt the cell cycle in the G0/G1 phase via cyclin D1 and cIAP2 regulation, and induce apoptosis without promoting necrosis via caspase-8/-3 and PARP cleavage. Constitutive nuclear factor-κB (NF-κB) signaling has been shown to contribute to the heightened inflammatory state and survival in TNBC cells. Herein, we also provide evidence that LOE inhibits NF-κB signaling by reducing RIP1 protein levels in MDA-MB-231 cells. These studies reveal that LOE suppresses key features of the progression of aggressive breast cancer cells and provides a basis for further definition of its underlying mechanisms of action and anticancer potential.

Spatiotemporal heterogeneity in precipitation patterns explain population-level germination strategies in an edaphic specialist.

BACKGROUND AND AIMS: Many locally endemic species in biodiversity hotspots are restricted to edaphic conditions that are fixed in the landscape, limiting their potential to track climate change through dispersal. Instead, such species experience strong selection for germination strategies that can track suitable conditions through time. Germination strategies were compared among populations across the geographic range of a California vernal pool annual, Lasthenia fremontii Local germination strategies were tested to determine the associations with geographic variation in precipitation patterns. METHODS: This study evaluated patterns of seed germination, dormancy and mortality in response to simulated variation in the timing, amount and duration of the first autumn precipitation event using seeds from six populations that span a geographic gradient in precipitation. Next, it was tested whether the germination strategies of different populations can be predicted by historical precipitation patterns that characterize each site. KEY RESULTS: A significant positive relationship was observed between the historical variability in autumn precipitation and the extent of dormancy in a population. Marginal populations, with histories of the most extreme but constant autumn precipitation levels, expressed the lowest dormancy levels. Populations from sites with historically higher levels of autumn precipitation tended to germinate faster, but this tendency was not statistically significant. CONCLUSIONS: Germination in L. fremontii is cued by the onset of the first rains that characterize the beginning of winter in California's Great Central Valley. However, populations differ in how fast they germinate and the fraction of seeds that remain dormant when germination cues occur. The results suggest that seed dormancy may be a key trait for populations to track increasingly drier climates predicted by climate change models. However, the low dormancy and high mortality levels observed among seeds of the southernmost, driest populations make them most vulnerable to local extinction.

Intensity and duration of intermittent exercise and recovery during a soccer match.

Soccer is a sport consisting of high-intensity intermittent exercise, with players making forays across their anaerobic threshold for tactical advantage followed by periods of recovery. The intensity and duration of these work and recovery bouts were defined during a men's soccer match using StepWatch Activity Monitors recording step rate for each 3-second period. The data were coded by custom software to separate work bouts (step rate ≥ 4) from recovery bouts (step rate < 4), and a square wave of the pattern of bouts was plotted for 5 players: center forward, central midfielder, wing midfielder, central defender, and wing defender. Four values were calculated for each work and recovery bout identified: duration, and mean, maximum, and minimum step rate (intensity). This novel technique provided detailed graphical information on the duration and exercise intensity of each position throughout the match. The center midfielder was able to sustain work and recovery bout characteristics throughout the match and appeared to recover at higher intensity levels than other players. The forward showed the consequence of accumulated fatigue late in the match and was unable to sustain the duration of high-intensity work bouts observed earlier in the match. The central defender attenuated the intensity of his work and recovery bouts late in the match staying closer to a more moderate work rate with fewer high- or low-intensity bouts. Having objective data qualifying players' work and recovery bout characteristics might prove valuable for tactical decision making, substitution timing, and for planning future training sessions.

Kinematic assessment of treadmill running using different body-weight support harnesses.

10 male collegiate runners (M age = 21.4, SD = 1.5 yr.) ran on a treadmill with no body-weight support (BWS), 20% BWS, and 40% BWS conditions. In addition, they wore three different commercially available harnesses at the 20% and 40% BWS conditions. The aim was to run on the treadmill at a fast speed while maintaining an adequate step length. The purpose was to investigate how each harness changed running gait, and the differences in running gait between the harnesses with various body-weight support. Analysis of variance indicated significant restriction of upper body torso rotation between the harnesses at the 40% BWS conditions. Body-weight support resulted in a longer stride, decreased cadence, less vertical displacement of the center of mass, and diminished hip and ankle joint excursions. These changes indicated that increased body-weight support results in longer steps with the foot contacting the belt for a shorter period of time with less leg angular changes throughout the running cycling.

Comparative Kinematic Measures of Treadmill Running with or without Body Weight Support in Runners.

Treadmill walking and running using a supportive harness has been used as a training method to rehabilitate injured patients' walking or running gait. Comparison of full weight support (FWS) and body weight support (BWS) kinematic measures in competitive runners has received little attention. The purpose of this study was to compare selected FWS to BWS kinematic measures in healthy competitive runners. Ten male runners (age = 21.4 ± 1.5 years) with a training regimen averaging 64 km per week at 3.8 m·s(-1) participated. All participants ran three 3-minute trials. The randomized trial conditions were: FWS, 20% BWS, and 40% BWS. All conditions were videotaped with 2 cameras and a 21-point, 3-D model was generated for analysis. From the position-time data, cycle length (CL), cycle frequency (CF), time of contact (TC), hip-, knee-, ankle- range of motion in degrees (H-ROM, K-ROM, and A-ROM, respectively), and vertical displacement of the center of mass (COM) were derived and compared. With increasing support conditions, cycle length increased. Cycle frequency, hip and ankle angle ranges, and COM vertical displacement decreased (p < 0.05). However, the ankle angle range of motion varied little between 20% and 40 % BWS (p > 0.05). BWS running produced significant changes in selected kinematic measures. These changes may provide insight into runners' behavior when using BWS in training or recovery from competition. Additional investigation of BWS training affect with competitive runners would be recommended. Key PointsRunning kinematics change gradually with increasing levels of body weight support.At constant speed, center of mass vertical displacement, lower leg angular displacement, foot contact time and cycle frequency decrease significantly from FWS to 40S.Harness running can have beneficial effects in healthy runners recovering from intense bouts of exercise.