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Comarca Ngöbe-Buglé Tour

Guided Tour through Indigenous Village, including Oreba Chocolate Farm Visit

Visit the Ngöbe-Buglé´s organic cocoa farm, part of the richness of this indigenous group’s customs and culture that forms an essential part of their economy. The Ngöbe-Buglé will guide you through their traditional preparation of cocoa and discuss its’ importance to their daily lives. Through dance, artwork, food, language lessons and tour of their home, including the surrounding waterfalls, trails and plantations, experience the richness of their hard-fought traditions and independence. With the option to horseback ride, purchase of handmade necklaces and hats, and the scenic views of their lush green home, your morning adventure will be both eye-opening and full of love and appreciation for a beautiful way of life.

Includes: bilingual guide, hotel pick-up/drop-off, lunch, refreshments


“My interview with Hope Cell founder Robert Vancina provides an insight not only into his breakthrough technology but a glimpse of the future where zero carbon footprint will go hand-in-hand with mobile energy storage and delivery.

The last hurdle to the large-scale adoption of fuel cells is about to fall, with the green and efficient production of hydrogen provided by Hope Cell.” Jurgen Vollrath Exponential Technologies Counsel

Application Carbon

Methane gas application

The present invention relates to plasma decomposition of methane and other hydrocarbons for producing hydrogen through plasma reforming process.

Excerpt from patent papers: ” An aspect of the invention is an improved apparatus and method for providing stable and controllable plasma for the purpose of generating hydrogen via plasma decomposition of hydrocarbon materials. Employing plasma, through proposed method and apparatus, as medium for changing state of the matter of the hydrocarbon enclosed in the vacuum, change physical properties of the cluster. Plasma electrical charge ionizes hydrocarbons and enables lower temperatures of hydrocarbon decomposition through resonating bonds in the cluster using hydrogen exothermic effect with a highly energetic rate and resulting in more effective breakage of hydrogen-carbon bond. This approach gives a number of advantages: lower energy consumption, higher energy efficiency in production, starting and stopping the decomposition process is close to instantaneous, user-friendly control with possibility of instant variable output of the process, scalability of application, decomposition approaching 100% under optimal high pressure, low electrical conductivity of input hydrocarbon gas can be converted through plasma discharge into high conductivity physical properties.

Methane US011 Certificate of Grant

A wide variety of hydrocarbon compounds can be used in plasma decomposition according to this invention where carbon, as the by-product is released in solid soot state and is easily removable and ready for usage in different applications or safe storage. Important characteristics of the process are simplification of the decomposition, no need for catalyst so no catalyst deactivation, scalable size, on-demand usage, mobile equipment friendly and low-cost applications.”

And just an observation: side effect – Atomic Carbon sot as a perfect source for graphene nanotechnology feedstock

One of the main problems associated with hydrogen production in hydrocarbon conversion process, from a chemical point of view, is the kinetic limitation. Low feasibility narrowing options of the process for conventional thermal conversion. Most common production of hydrogen – Steam Reforming resulting with high energy consumption. Using special high-priced catalysts to attain reasonable productivity and equivalent equipment size without much scalability rate characterize this technology. The necessity to heat the catalyst to the high working temperature (approx.800 0C) leads also to the problem of ‘cold start’ and restricting mobile applications.  Enormous energy waste in the process accompanied with the production of over 8.8 billion tons* of carbon globally, are additional negative aspects of the process. Hydrogen basic physical properties ensure future wide usage as an energy source and carrier of high caloric value. A wide variety of applications can be adapted to hydrogen use as the source or medium of energy. Hydrogen is a very reactive element and does not exist in the elementary form in the natural environment of the Earth. It always comes in a molecular arrangement of clusters based on H2 dipole. Stability of those clusters depends on the stability of all elements included. Hydrogen is bonded with other elements not only as single-molecule bond but rather as oscillating clusters of molecules bonded together. Hydrogen bonds on photo and schematic                                                                                Hydrogen cluster image and schematic of it

The substantial cooperative strengthening of the hydrogen bonds is dependent on long-range interactions and strength of each bond in the cluster, which encourages larger clusters formation for the same average bond density and potential.

Elements isolation can be achieved by exposing cluster to a range of high temperatures. An unstable elementary hydrogen in the cluster, whose bond with other elements has been broken when exposed to high temperatures, will tend to react with predominantly electrically opposite element in its proximity. In a vacuum environment, it will form a hydrogen molecule.

Breaking one bond, through exposing cluster to heat, generally weakens those around. If exposed to the oxygen environment, and accompanied with high temperature, hydrogen will violently react in combining with oxygen through combustion. This mechanism would define most common combustion in general, allowing for some untypical exceptions. Exothermic reaction further breaks the hydrogen bond with other elements of the cluster, exposes more hydrogen to run off combustion process.

If we take, for example, hydrocarbon case, different hydrocarbon bonds occur in various lengths and structures, comprise various additional elements as well. More complex hydrocarbon cluster can be broken to as many simple hydrocarbons and other components through exposing to different temperatures.

Reactivity of metal hydride with hydrogen is known and used in various applications.

The phenomenon of hydrogen embrittlement results from the formation of interstitial hydrides. Interstitial hydrides most commonly exist within metals or alloys more closely resembling common alloys. In such hydrides, hydrogen can exist as either atomic or diatomic entity. Mechanical or thermal processing, such as bending, striking, or annealing may cause the hydrogen to precipitate out of solution, by degassing. These systems are usually non-stoichiometric, with variable amounts of hydrogen atoms in the lattice. Hydrides of this type form according to either one of two main mechanisms. The first mechanism involves the adsorption of dihydrogen, succeeded by the cleaving of the H-H bond, the delocalization of the hydrogen’s electrons, and finally, the diffusion of the protons into the metal lattice. The other main mechanism involves the electrolytic reduction of ionised hydrogen on the surface of the metal lattice, also followed by the diffusion of the protons into the lattice. The second mechanism is responsible for the observed temporary volume expansion of certain electrodes used in electrolytic experiments.

Those mechanisms do not have any typical side effects of an atomic reaction, supported by strong evidence of lattice transmutation through spectrometry readings, and can’t be considered as such.

Mechanism initiated through plasma treatment of hydrogen based cluster in presence of metal hydride lattice would present new moment in hydrogen embrittlement and is accompanied with exothermic reaction.

Hope Cell initial sea water test with LENR surface etching evidence; plasma relocation example

Practical example of hydrogen embrittlement through proposed mechanism in metal hydride lattice

Plasma is a highly dense source of energy, which covering process enthalpy and provide optimal temperature range to eliminate kinetic limitations of hydrogen isolation.

Low electrical conductivity of the medium has been converted into high conductivity physical properties through the interaction of plasma which resulting in a change of the state of the matter.

Double Layer plasma mechanism isolates an unstable and highly reactive elementary atomic hydrogen H in the cluster, whose bond with other elements has been broken. Exposed atomic hydrogen proton will violently react with surrounding fast moving metal hydride lattice electron and forming additional neutron through isolated but violent exothermic reaction. This additional exothermic reaction – highly energized emission, results in elevated atomic hydrogen isolation by syncing into molecular dipole frequency with resonating effect, where excessive breakage of surrounding cluster bonds is maintained in a run-off process. Breaking one bond, through exposing hydrogen medium cluster to excess heat, bends and weakens bonds around, and a process is repeated in surrounding area of metal hydride lattice. Mechanism results in forming of 2H Deuterium, which is one neutron heavier, and sheds excess binding energy to the lattice through beta decay, further resulting in the nano-dimensional isolated transmutation of the surface, with spectrometry detection of numerous new elements. Plasma electromagnetic excitement allows the process to continue with the hydrogen proton capture in the lattice. Each successive cascade and decay emit a significant amount of excess heat energy and result in further isolated metal hydride surface lattice transmutation through this weak nuclear force.

Hydrogen Proton

Schematic of Hydrogen embrittlement in metal hydride lattice and conditions of exothermal reaction mechanism LENR

Hydrogen based cluster decomposition through double layer plasma mechanism demonstrates a high specific productivity rate of decomposition comparing with steam reforming or partial oxidation processes.

Employing plasma, as a medium for changing the state of the matter of the cluster, changes its physical properties. Plasma electrical charge ionising hydrocarbon and allowing lower temperatures of decomposition from approximately 8000C in conventional steam reforming to approximately 1200C with plasma decomposing through resonating bonds in the cluster with a high energetic rate. The process resulting in more effective and substantially less energy demanding breakage of a hydrogen-carbon bond. Enthalpy of the mechanism covering a wide range of temperatures where different hydrogen based clusters can be decomposed. The process demonstrates over-unity comparing to electrolysis or steam reforming and is proportionally reflected by lowering final price of the product.Hope Cell Double Layer Plasma Mechanism

This approach gives numeral advantages of Lower energy consumption; Higher energy efficiency in production; Starting and stopping process of decomposition close to instantaneous; User friendly control with possibility of instant variable output of the process; Scalability of application; Decomposition approaching 100% under optimal high pressure; Wide variety of hydrogen based cluster compounds can be used in plasma decomposition through proposed method, where carbon, as the by-product is released in solid soot state – it is easily removable and ready for usage in different applications or safe storage. An important characteristic of the process are the simplification of the decomposition; no need for catalyst so no catalyst deactivation; scalable size; on demand usage; mobile equipment friendly; low-cost applications. Water decomposing would be the most obvious application as well.

  magnyfied plasma thermoionic scatering under 720 Wh

Result of applying mechanism to sea water decomposition

Uniquely scalable setup allowing the exothermic effect of hydrogen in a robust stainless steel enclosure with LENR evidence; Neutron capture and weak interactions explain the surface reactions and excess heat generation. Hope Cell have surface interactions spread throughout the body of the cell on multiple rates – example of the discovery of controlling, directing and magnification. Process strongly supporting water dislocation in anomalous over-unity quantity comparing to standard electrolysis. Burned mark on another side of the body showing plasma change of the state of the matter of the water and physical properties as a result of it (water can burn)! Using surplus of the wind, sun-generated energy for conversion to hydrogen for readily available, on demand usage is another innovative example of converting hydrogen to medium or carrier of energy, allowing alternative sources to become mainstream as a major breakthrough in energy consumption. There are many more exciting possibilities.

* Source – Royal Society of Chemistry 2009

Journal of Advanced Chemical Engineering

High Impact List of Articles

Journal of Fundamentals of Renewable Energy and Aplications Publication

Published October 2015

Research Gate Publication

Published July 2014

Naval Surface Warefare Center Featuring Hope Cell

Published September 2015

More info -Web<a href=&quot,http://www.energyglobe.com/en/energy-globe-award/press-room/press-photos/recent-photos/national-winner-prague-09/continent-europe/

WIPO 1 (1)WIPO 1 (2)US Patent012US Patent No1 Methane US011 australian-patent 1

Certificate of Grant

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How much are those apple fritters going to ‘cost’ you in calories?

So you indulged in a pair of apple fritters. (No one is blaming you.) But now you want to undo the damage.

Each fritter is about 300 calories, according to Here are some ways a 145-pound person could burn off 600 calories. The precise number of calories a person burns are influenced by the person’s age, body weight, gender, activity level and movement efficiency. Use this only as a guide.

canoe3 hours, 2 minutes of leisurely canoeing

2 hours, 46 minutes of walking at 3.4 miles per hour on a flat surface

2 hours of vigorous housecleaning

1 hour, 2 minutes of cycling at 12 to 13.9 miles per hour on a flat road

50 minutes of vigorous lap swimming

–reviewed by exercise physiologist Carol Sames, PhD, director of the Vitality fitness program at Upstate’s Institute for Human Performance.

This entry was posted in community, diet/nutrition, entertainment, fitness. Bookmark the permalink.


PLoS ONEplosplosonePLoS ONE1932-6203 Public Library of Science San Francisco, USA 08-PONE-RA-07797R1-A10.1371/journal.pone.0005298Research ArticleEcology/Environmental MicrobiologyMicrobiology/Environmental MicrobiologyMicrobiology/Microbial Evolution and GenomicsModern Subsurface Bacteria in Pristine 2.7 Ga-Old Fossil Stromatolite Drillcore Samples from the Fortescue Group, Western AustraliaBacteria in Old Stromatolites GérardEmmanuelle1 MoreiraDavid2 PhilippotPascal1 Van KranendonkMartin J.3 López-GarcíaPurificación2* IPGP-IMPMC, Université Denis Diderot et CNRS, Paris, France Unité d'Ecologie, Systématique et Evolution - CNRS UMR8079, Université Paris-Sud 11, Orsay, France Geological Survey of Western Australia, East Perth, Western Australia, Australia WardNaomi Editor University of Wyoming, United States of America * E-mail:

Conceived and designed the experiments: DM PLG. Performed the experiments: EG. Analyzed the data: EG DM PLG. Contributed reagents/materials/analysis tools: PLG. Wrote the paper: EG PLG. Sampling processing: PLG DM. Co-organised the Pilbara Drilling Project: PP MJVK.

The authors have declared that no competing interests exist.

Several abiotic processes leading to the formation of life-like signatures or later contamination with actual biogenic traces can blur the interpretation of the earliest fossil record. In recent years, a large body of evidence showing the occurrence of diverse and active microbial communities in the terrestrial subsurface has accumulated. Considering the time elapsed since Archaean sedimentation, the contribution of subsurface microbial communities postdating the rock formation to the fossil biomarker pool and other biogenic remains in Archaean rocks may be far from negligible.

Methodology/Principal Findings

In order to evaluate the degree of potential contamination of Archean rocks by modern microorganisms, we looked for the presence of living indigenous bacteria in fresh diamond drillcores through 2,724 Myr-old stromatolites (Tumbiana Formation, Fortescue Group, Western Australia) using molecular methods based on the amplification of small subunit ribosomal RNA genes (SSU rDNAs). We analyzed drillcore samples from 4.3 m and 66.2 m depth, showing signs of meteoritic alteration, and also from deeper “fresh” samples showing no apparent evidence for late stage alteration (68 m, 78.8 m, and 99.3 m). We also analyzed control samples from drilling and sawing fluids and a series of laboratory controls to establish a list of potential contaminants introduced during sample manipulation and PCR experiments. We identified in this way the presence of indigenous bacteria belonging to Firmicutes, Actinobacteria, and Alpha-, Beta-, and Gammaproteobacteria in aseptically-sawed inner parts of drillcores down to at least 78.8 m depth.

The presence of modern bacterial communities in subsurface fossil stromatolite layers opens the possibility that a continuous microbial colonization had existed in the past and contributed to the accumulation of biogenic traces over geological timescales. This finding casts shadow on bulk analyses of early life remains and makes claims for morphological, chemical, isotopic, and biomarker traces syngenetic with the rock unreliable in the absence of detailed contextual analyses at microscale.

This work was supported by ATIP (PLG), Programme National de Planétologie (PP) and Geomex (PLP and PP) grants of the Centre National de la Recherche Scientifique, and by the Agence National de la Recherche (PP and PLG). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

In recent years, there has been an intense debate about the authenticity of the oldest claimed traces of life in ancient sedimentary rocks, as most of them, including the accumulation of light isotopes, the presence of organic compounds or that of microfossil-like precipitates, could be simply the product of abiotic processes, particularly those occurring under hydrothermal conditions [1], [2], [3]. The presence of fossil lipids in Archaean rocks cannot be the product of abiotic synthesis and, until recently, was generally accepted as unambiguous evidence for the presence of specific groups of organisms at the time when the sediment deposited. However, ancient life signatures in general might also be the product of later contamination. A recent example highlights this potential problem. In 1999, Brocks and colleagues reported the presence of 2α-methylhopanes and steranes in 2.7 Ga-old rocks from the Pilbara Craton (Australia) that were formerly attributed to 2.7 Gy-old cyanobacterial hopanoids and eukaryotic sterols, respectively [4], [5]. However, recent isotopic δ13C values measured at microscale of pyrobitumen and kerogen within the same samples analyzed by Brocks et al. (1999) showed that these are much lighter than values for previously extracted hydrocarbons, suggesting that the latter were later contaminants and reverting the oldest fossil evidence for cyanobacteria and eukaryotes to 2.15 and 1.78–1.68 Ga-old, respectively [6].

There are in principle three potential sources of post-syngenetic contamination by fossil biomarkers and other types of biogenic traces: i) contamination of rock cores during drilling if hydrocarbon-rich oil or kerosene, which also carry isotopic and other life signatures, are used in drilling fluids, ii) migration of fluids containing biogenic signatures to rock layers initially devoid of them, and iii) the accumulation of biomarkers produced in situ by microorganisms inhabiting the rock after its formation. Whereas the first two possibilities have been often considered [7], the latter is generally ignored despite the accumulation of a large body of evidence showing the occurrence of diverse and active microbial communities in the terrestrial subsurface [8], [9], [10], [11], [12]. Here, we contend that, considering the time elapsed since Archaean sedimentation, the contribution of subsurface microbial communities postdating the rock formation to the biogenic signature pool may be far from negligible and that, in some instances, might even preclude hope to discriminate between truly ancient signatures and posterior contaminants.

In order to determine if well-preserved subsurface ancient sedimentary rocks potentially harboring microfossils or other biogenic traces could be considered sterile and exempt of late, post-syngenetic, microbial remains, we investigated by molecular methods based on the amplification SSU rDNAs a suite of pristine diamond drill cores (PDP1 drillcore samples) intersecting Archaean stromatolitic layers that had been collected during the Pilbara Drilling Project from the 2,724 Myr old Tumbiana Formation (Fortescue Group, Hamersley Basin) at Meentheena, Western Australia [13]. The rocks of this and immediately above formations harbor well-preserved fossil stromatolites and have provided a wealth of biogenic signatures, including very negative δ13C values presumably linked to Archaean methanotrophic activity [14], filamentous microfossils [15], and the above-mentioned 2α-methylhopanes and steranes that were suggested to derive of 2.7 Gy-old cyanobacterial hopanoids and eukaryotic sterols [4], [5], although not without controversy [6]. Our molecular diversity studies reveal the occurrence of indigenous bacteria in aseptically-sawed inner parts of drillcores of the Tumbiana Formation down to at least 78.8 m depth, which has important implications for the interpretation of the ancient fossil record.

Results and Discussion

To look for the putative presence of contemporary microorganisms in 2.7 Ga-old stromatolites, we selected samples from different lithological layers in the field (4.3 m, 66.2 m, 68.0 m, 78.8 m and 99.3 m depth) immediately after the PDP1 core was drilled (Figure 1), which were subsequently treated with maximum precautions to avoid external contamination during sawing and grinding, DNA extraction and subsequent PCR experiments (see Methods). The two upper samples showed evident signs of alteration. The 4.3 m-deep core sample consisted of altered subaerial basalt, and the 66.2 m-deep sample of pervasively weathered reddish stromatolites, which indicated mineral oxidation linked to water circulation, the water table being present approximately until that depth. Deeper samples were apparently unaltered and showed no macroscopic signs of channeled or pervasive fluid infiltration. The 68.0 and 78.8 m-deep samples corresponded to well-preserved dark grey stromatolites consisting of alternated carbonate and mudstone layers. The 99.3 m-deep sample was composed of volcanogenic tuffaceous material [13] (Figure 1).

10.1371/journal.pone.0005298.g001 Samples analyzed from the Pilbara Drilling Project drillcore PDP1.

Black triangles indicate the regions in the schematic lithological PDP1 log [13] where samples for microbiology analyses were retrieved from. Photographs showing the samples analyzed are shown on the left. The scale bar corresponds to 1 cm.

DNA was extracted from 0.5 g ground core fragments. In the case of stromatolitic layers, (66.2 m and 68.0 m), we performed an additional purification from 10 g of material. To limit the effect of the possible introduction of exogenous microorganisms by the drilling fluid in cores, we sawed them aseptically and retained exclusively the inner part for molecular analyses in the case of the deeper, unaltered samples (68.0 m, 78.8 m and 99.3 m). In addition, to evaluate potential contamination by the drilling fluid, we i) extracted DNA from the drilling fluid used to establish a catalogue of potential drilling-fluid contaminants and ii) analyzed both, inner and outer core parts of the 66.2 m-deep sample. We made no distinction between the inner and outer part for the 4.3 m, which we considered as positive control since, being a shallow altered sample, it might contain more biomass than deeper samples. To establish additional controls of putative laboratory and/or DNA extraction kit contaminants, we subjected blank tubes to the DNA extraction protocols along with drillcore samples and we also extracted DNA from the remaining fluid once the drillcores were sawed. Although the sawing step was a potential source of contamination of core samples, sawing-fluid associated microorganisms may not necessarily be contaminants, but indigenous organisms released from the cores during sawing.

We amplified SSU rRNA genes from drillcore and drilling and sawing fluid purified DNA. Weak amplification bands were obtained for 4.3 m, 66.2 m and 68.0 m levels, but in too low amount to construct SSU rDNA libraries for molecular diversity purposes. Nested PCR reactions were then carried out to obtain clear amplification bands that served to construct SSU rDNA libraries (Table 1). Being concerned by exogenous contamination problems, we additionally made control SSU rDNA libraries of drilling and sawing fluids as well as laboratory-associated contaminants. The latter comprised 2 libraries of blank control tubes from the DNA extraction kit as well as 6 additional libraries derived from control reactions in nested PCR experiments (controls in direct PCR experiments were all negative) (Table 1). In most cases, we sequenced all the clones that were generated in the different drillcore libraries and, in the case of drilling fluid and laboratory controls, we sequenced clones until a plateau indicating complete coverage of sample biodiversity was reached or nearly so (Figure 2). Based on the list of laboratory and drilling fluid potential contaminants and also in other lists of contaminants in the literature [16], [17], we classified PDP1 phylotypes along a gradient according to their likelihood to be contaminant or truly indigenous (Figures 3 and 4, and Table S1).

10.1371/journal.pone.0005298.g002 Rarefaction curves of drillcore and control SSU rDNA libraries.

The number of operational taxonomic units (OTUs) accumulated can be observed as a function of the number of total sequences obtained. OTUs were defined as clusters of SSU rDNA sequences >97% identical.

10.1371/journal.pone.0005298.t001 SSU rDNA amplification, libraries generated, and clones yielded and/or analyzed in PDP1 drillcores of different depths.
Direct PCR Nested PCR Number of OTUs
Samples PCR product No. libraries produced No. clones obtained/analyzed PCR product No. libraries produced No. clones obtained/analyzed
Drill core samples
4.3 m +/− 2 17 + 3 63 2
66.2 m outer +/− 1 3 + 4 147 7
66.2 m inner +/− 1 8 + 6 308 12
68.0 m inner +/− 1 4 + 5 244 3
78.8 m inner + 4 173 5
99.3 m inner + 5 155 3
Control samples
Sawing fluid +/− + 3 56 5
Drilling fluid + 7 317 nd nd nd 20
Laboratory* + 8 367 19

(PCR+DNA extraction kit) associated contaminants

nd not done

OTU, operational taxonomic unit.

Laboratory contaminants coming from the surrounding environment, PCR reagents or the DNA extraction kit accounted for a relative high proportion of sequences in SSU rDNA libraries (Figure 3) belonging mostly to Gram positive bacteria and various subdivisions of the Proteobacteria (Figure 4). Most of them affiliated to the genera Sphingomonas, Stenotrophomonas, Ralstonia, to a divergent alphaproteobacterium (clone 4_60) and to Shigella/Escherichia, the latter likely deriving from trace amounts of E. coli DNA in recombinant Taq polymerase aliquots as it was associated to PCR controls, and they comprised 36%, 16%, 11%, 11%, and 8% of the total contaminant clones, respectively. Although not found in laboratory control libraries, we also discarded clones that shared more than 95% sequence identity with animal or human-related bacteria as potential contaminants (Figure 4 and Table S1). Contaminants introduced by the drilling fluid comprised moderate proportions that were observed in all the drillcore samples down to 99.3 m depth. The proportion of drilling fluid-associated phylotypes was not significantly higher in the outer part of the 66.2 m-deep core compared to its inner part and was comparable to that of the 4.3 and 68.0 m-deep layers (Figure 3). This suggests that, the porosity of the different samples being very low [18], contamination by the drilling fluid was not extensive in the rock and that contamination of both, inner and outer part of cores, occurred through microfractures. Phylotypes identified in the drilling fluid affiliated to the Sphingomonadaceae family (48% of the total drilling fluid clones) or were related to the genera Delftia (24%), Agrobacterium (14%) and, distantly, Flavobacterium (14%). Although we considered them as contaminants introduced by the drilling fluid, microorganisms belonging to the Sphingomonadaceae family or to Delftia species are ubiquitous, being frequently isolated from aquifers [19], [20], and the possibility that they might be found not only in the water used for drilling but also in the water circulating in the subsurface cannot be completely excluded. This caution might apply also to some phylotypes that we have considered here as laboratory contaminants. For instance, we identified a phylotype closely related to Hydrogenophilus spp. (Figure 4, Table S1), moderately thermophilic hydrogen and sulfur-compound oxidizers [21] associated to subsurface aquifers, oil wells and even, perhaps, the lake Vostok [22]. Although we consider it here as a contaminant, Hydrogenophilus relatives are also present in the subsurface aquifer of the Great Australian Artesian Basin [23], and might therefore be actually present in PDP1 samples. Phylotypes detected in the sawing-associated fluid were not considered as indigenous subsurface microbes and were classified apart, although the possibility that they come from the core samples themselves cannot be discarded.

10.1371/journal.pone.0005298.g003 Relative abundance of contaminant and indigenous SSU rDNA sequences in the different PDP1 drillcore sample libraries. 10.1371/journal.pone.0005298.g004 Maximum likelihood phylogenetic tree showing the position of different OTUs identified in PDP1 drillcore and control (laboratory, drilling fluid, sawing fluid) samples.

The tree was constructed including the closest possible cultivated members to our sequences. The number of phylotype counts in SSU rDNA libraries from different samples is given on the right. Phylotypes are color-coded attending to their classification along a gradient going from indigenous and potentially indigenous to laboratory contaminants. Clones considered indigenous or potentially indigenous were found exclusively in PDP1 drillcores, having no close relatives (>95% identical) in control contaminant libraries or in the sawing fluid-associated library. Only bootstrap values above 50% are given at nodes.

Phylotypes found in drillcores that were not detected in control libraries but that were found only once or belonged to ubiquitous lineages having relatives identified also in contaminant libraries from literature data were considered only as potentially indigenous. This was the case of several alphaproteobacterial sequences shared by samples coming from 66.2 m to 78.8 m depth that formed 3 OTUs affiliated to the genus Sphingomonas or Sphingobium. Species of these genera are ubiquitous aerobic, chemoheterotrophic bacteria having been isolated from various environments including the deep subsurface [19], [20], [24], [25] but also clean rooms where space craft are assembled [17]. Similarly, various sequences forming three OTUs affiliated to the genus Methylobacterium were considered only as potentially indigenous (Figure 2 and Table S1). Although members of this genus have been detected as contaminant in some libraries in the literature [26], their distribution is widespread in many different environments, and moderate thermophilic Methylobacterium-like alphaproteobacteria have been isolated from deep aquifers in Australia [27].

Finally, we classified as truly indigenous to the Tumbiana subsurface, sequences found in general at several core depths or retrieved from independent clone libraries. Several of these phylotypes were more or less distantly related to Alpha-, Gamma- and Betaproteobacteria, Firmicutes and Actinobacteria sequences identified in soil but also in hot springs or subsurface sediments (Figure 4 and Table S1). One Firmicutes phylotype was distantly related to Bacillus and Paenibacillus sequences retrieved from subsurface sediments [28]. Iron and manganese reducing Bacillus species have been isolated from deep aquifers in Australia [29]. We also detected phylotypes closely related to the alphaproteobacterium Pedomicrobium australicum in samples 66.2 and 68.0 m. Pedomicrobium species are manganese and sometimes iron oxidizing bacteria able to colonize a wide variety of environments including desert rocks [30], [31]. Some of them can also metabolize methanesulfonic acid, a capability also shared by Methylobacterium species [32], and sulfur-containing aromatic compounds [33], playing also a role in the sulfur cycle. Since iron, manganese and sulfur compounds are abundantly available in these Archaean rocks, they might be used as electron donors by a variety of subsurface microorganisms. The other two indigenous alphaproteobacterial phylotypes identified were only very distantly related to any described species or environmental sequences, and might thus represent new lineages of subsurface-associated microorganisms. This was also the case of Betaproteobacteria indigenous phylotypes, which were only distantly related to environmental sequences retrieved from geothermal areas or compost. Among the Gammaproteobacteria, we identified three phylotypes indigenous to the drillcores. One of them was closely related to Silanimonas lenta, isolated from a hot spring, and to a Xanthomonadaceae sequence (Acc.No. DQ230964) retrieved from subsurface water in Kalahari, South Africa.

The fact that we amplified indigenous bacterial DNA in our samples did not prove that these bacteria are actually alive and active. They might be dormant or the detected DNA could correspond to the remains of dead bacterial cells. However, DNA is a relatively unstable molecule that may be rapidly degraded after the death of the microorganisms, first by endogenous nucleases and, then, by chemical processes like hydrolysis or oxidation [34]. In certain conditions of desiccation or high salinity, DNA may be more stable and several studies have claimed that bacterial DNA could survive more than 10 millions years [35], [36], [37], [38] but, in addition of being highly controversial claims [39], [40], a few million years is a short time in the geological record, especially when 2,724 Myr old Archaean rocks are being considered. In addition, fossil DNA is characterized by being split in small fragments by the above-cited hydrolytic processes. Since we amplified 1465-bp long DNA sequences, it is highly improbable to obtain such long amplification from fossil DNA older than few thousand years [41]. Furthermore, several of our indigenous or potential indigenous candidate SSU rDNAs share up to 99% identity with those of other modern bacteria. If these sequences were truly derived from the amplification of Archaean bacteria, they should have diverged more that 1% at this locus since the deposition of these rocks some 2,724 Myr ago. Such low evolutionary rate at this locus is incompatible with current knowledge of gene evolution. We thus conclude that the DNA we detected in our core samples comes from modern bacteria that are alive or were still alive few thousand years ago.

How many contemporary cells might be present in Hamersley subsurface fossil stromatolites? This is very difficult to estimate due to a likely low cell density combined with the difficulties associated to the study of subsurface rock-associated bacteria. By direct PCR, we only detected weak amplification bands in some samples, in agreement with the presence of low biomass even in altered samples. Direct counting of bacterial cells using epifluorescence microscopy and DNA labeling with fluorochromes such as DAPI or Syto9 was not possible. Carbonate-rich mineral samples exhibit high autofluorescence and have affinity for usual DNA stains used to detect microbial cells, making it very difficult to distinguish bacterial cells from mineral particles in carbonate rocks, particularly in samples where the biomass is extremely low, as was likely the case. The combination of carbonate autofluoresce and low cell density prevents also the use of metabolic dyes that target intact cell membranes such as the LIVE/DEAD® Viability kit (Molecular Probes™) or that of fluorescent in situ hybridization (FISH) experiments using specific probes, making it extremely difficult to distinguish between artefactual binding from true cell labeling. This kind of detection is further impeded if we consider that cells present in the rock may be dormant or have extremely low metabolic rates. This would imply that metabolic staining may be below detection limits. Similarly, cells with low or no detectable metabolic activity may have too few ribosomes for FISH probes to produce a detectable signal. Therefore, we cannot provide precise estimates of cell numbers in our samples but the weak amplification signal obtained in most PCR experiments suggests that cell density is probably very low. For comparison, microbial cell densities of ca. 102–104 cells/g rock have been estimated in ultradeep mines [16] and other continental drilling samples [12], [42].

Our results strongly suggest that contemporary bacteria inhabit what are generally considered exceptionally well-preserved subsurface Archaean fossil stromatolites of the Hamersley Basin, Western Australia. They are possibly in very low numbers, their distribution confined to microfractures where water may circulate (perhaps only intermittently), and their metabolic activities might be extremely low. However, upon geological timescales spanning 2.7 Gy, even such low cell numbers must have contributed significantly to the pool of biogenic signatures associated to these rocks, including microfossils, biological isotopic fractionation and lipid biomarkers. Although our results do not necessarily invalidate previous analyses, they cautiously question the interpretation of ancient biomarkers or other life traces associated to old rocks, even pristine, as syngenetic biogenic remains when bulk analyses are carried out. In this context, the development of high-resolution techniques that allow the identification of biogenic signatures in microscale-preserved environments is particularly promising. For example, high resolution spectroscopy and microscopy analyses revealed the presence of aragonite nanocrystals closely associated with organic nanoglobules, supporting a microbial role for stromatolite formation within well-preserved 68.2 m-deep stromatolitic layers in the same PDP1 drillcore that we have studied here [18], and measurements of carbon isotope deviations at microscale have allowed the discrimination between syngenetic organic matter and later biomarker contaminants in Archaean sedimentary rocks, eliminating false positives and paving the way for more accurate datation based on fossil biomarkers in the future [6].

Methods Sample collection and preparation

The samples analyzed in this study were obtained in August 17th and 18th 2004 from one diamond drillhole section (PDP1) that intersected one stromatolitic horizon through the c. 2.72 Ga Tumbiana Formation (Fortescue Group, Mount Bruce Supergroup, Hamersley Basin). Sampling was made possible within the framework of the Pilbara Drilling Project between the Institut de Physique du Globe de Paris and the Geological Survey of Western Australia. The detailed geological log of the diamond drillhole section has been reported [13]. Briefly, PDP1 intersected subaerial basalts of the Maddina Formation, stromatolitic carbonates and interbedded black shale of the Meentheena Member of the Tumbiana Formation, and underlying volcaniclastic sandstones. Stromatolites were identified by domical shapes and wrinkly laminations, including black organic-rich laminae. Local water from the Nullagine river was used for drilling. PDP1 was drilled with NQ2 core (47.6 mm diameter) from the surface to a depth of 104 m (Figure 1). Drillcores were extracted with maximum precautions to avoid external environmental contamination. All manipulation with bare hands by the operators of the Mount Magnet Drilling Company was prevented and cores were directly deposited in clean steel core trays. Digital photographs and lithological logs of the core were made in the field. Core sections selected for microbiological analyses were immediately separated using clean laboratory gloves and preserved in sterile plastic bags from external contact during transportation of core trays to the Geological Survey in Perth. Selected core fragments were sawed to extract the inner core (ca. 2 cm×2 cm×10 cm) for DNA extraction and analysis of microbial diversity. Prior to use the saw was treated extensively with 5% sodium hypochlorite for several hours and then rinsed with sterile mineral water. The saw reservoir was then filled with sterile mineral water and mixed with sodium hypochlorite to reach a diluted solution (ca. 0.1%). Core surfaces were cleaned with ethanol prior to sawing. During sawing the cores were hold with clean laboratory plastic gloves avoiding touching the inner core, which was finally collected using heat-sterilized forceps. Inner core surfaces were then pressed against a sterile gaze soaked with 5% sodium hypochlorite and the inner cores introduced in 50-ml Falcon tubes and fixed with 96% ethanol from a newly opened bottle. Treatment with 5% sodium hypochlorite was found to be the most efficient protocol for decontamination of external contaminating microbes and nucleic acids while maintaining those of internal microbes in ice cores [43]. Outer core sections were kept dry in sealed sterile plastic bags. After their transportation to the microbiology laboratory at the University of Paris-Sud, Orsay, France, all samples were stored at −20°C until use.

In addition, samples of the drilling fluid and sawing water used were also collected and fixed in situ to establish control SSU rRNA gene libraries of potential microbial contaminants introduced during the drilling process. The drilling fluid consisted of a mixture of river water with silicon oil. No drilling mud or diesel was put down the hole during drilling to avoid undue contamination by foreign organic material. The drilling fluid, which was at a pH of 7.0, was collected directly from the tank feeding the diamond drillcore during the drilling operation at Pilbara. 50 ml were filtered through a TMMP Millipore 5 µm-pore size filter and then through a GTTP Millipore 0.2 µm-pore size filter in a field laboratory set for this purpose. All the materials used were sterile. Upon filtration, the filter was fixed in ethanol for preservation and transport until the microbiology laboratory at Orsay, France. Filters were then stored at −20°C until use. Similarly, ca. 25 ml of the water used for core sawing containing a high density of rock particles in suspension was collected at the Geological Survey laboratory in Perth immediately after sawing, fixed with two volumes of 100% ethanol for transportation to France and, upon arrival, stored at −20°C.

DNA extraction, PCR amplification, and cloning

All sample manipulations carried out at the University of Paris-Sud, including DNA extraction and derived DNA-based protocols were done in a Biocap™ RNA/DNA hood (Captair, Erlab, France), a vertical laminar flow chamber specifically designed for the manipulation of samples with high contamination risk, equipped with a HEPA filter that guarantees 99,999% filtration efficacy for particles larger than 0,3 µm in diameter. Prior to use the chamber was UV-sterilized. All laboratory materials and solutions used were sterile (most often sterilized twice). Aerosol resistant pipette tips were always used to reduce the likelihood of external contamination. We selected inner core samples from 4.3, 66.2, 68.8, 78.8 and 99.3 m depth representing characteristic layers in the log (Figure 1). In addition, the outer core fragments of the 66.2 m-deep sample, corresponding to altered stromatolitic layers, was also analyzed (Table 1). Initially, inner cores were fragmented into pieces and ground finely in a sterile agatha mortar. 0.5 g of drillcore powder from each level was used for molecular diversity studies. Subsequently, due to the low yield of PCR and cloning steps for deep cores, we additionally used 10 g of powder from deep stromatolitic layers 66.2 m (altered) and 68.0 m (non-altered) inner core samples. 1.4 g of wet decanted powder from the sawing associated-fluid was used for DNA extraction, which was then mixed with the DNA extracted from a GTTP 0.2 µm pore-size filter where the potential microbial biomass from the remaining sawing water fixed in ethanol was filtered through. DNA of the different samples, including 0.2 µm pore-size filters used to collect the biomass from drilling and sawing fluids cut into pieces, was extracted using the PowerSoil DNA isolation kit (MoBio Laboratories) with minor modifications. DNA was resuspended in 50 µl of sterile 10 mM Tris/HCl, pH 8.5, and conserved at −20°C. As a laboratory control of potential contaminants, for each different DNA extraction done, a blank tube was subjected in parallel to the same extraction protocol applied to drillcore samples in order to construct control clone libraries from kit material and/or other laboratory-associated contaminants that might be introduced in stromatolite samples during DNA purification. Two laboratory control samples were produced in this way and subjected to PCR amplification protocols simultaneously to drillcore DNA samples.

Bacterial SSU rRNA genes were amplified by PCR using different combinations of the bacterial-specific primers 27F (5′-AGAGTTTGATCCTGGCTCAG) or 63F (5′-CAGGCCTAACACATGCAAGTC) and the prokaryote-specific reverse primers 1492R (5′-GGTTACCTTGTTACGACTT) or 1397R (5′-GGGCGGWGTGTACAAGGC). 5 µl of DNA purified as described above were used in 25–30 µl-volume PCR reactions using either GoTaq polymerase (Promega, France) or the hot-start Platinum Taq (Invitrogen), always opening new aliquots for each set of drillcore SSU rDNA amplification reactions. These were performed under the following conditions: 30 cycles (denaturation at 94°C for 15 s, annealing at 55°C for 30 s, extension at 72°C for 2 min) preceded by 2 min denaturation at 94°C, and followed by 7 min extension at 72°C. Direct PCR reactions were carried out with different primer combinations and, when applicable (Table 1), 1 µl of the first PCR reaction carried out with the most external primers (27F and 1492R) was then used in nested PCR reaction using the same conditions with primers 63F and 1387R. Negative controls were set for all PCR reactions, all of which were negative in direct PCR reactions. Several negative controls in nested PCR amplification experiments yielded amplicons, which were then cloned to contribute establishing a catalogue of laboratory and/or PCR reagent contaminants. In total, eight libraries of PCR-associated laboratory contaminants were constructed. From them, a relative high proportion of these clones yielded sequences closely related to Escherichia coli, suggesting that traces of DNA may persist in Taq polymerase commercial aliquots. Cloning was done using the Topo TA Cloning system (Invitrogen) following the instructions provided by the manufacturers. After plating, positive transformants were screened by PCR amplification of inserts using flanking vector primers and PCR products partially sequenced using either 1387R or 1492R (Cogenics, France). Table 1 is a summary of the PCR amplification results, SSU rDNA libraries constructed and the number of clones obtained and/or analyzed per library corresponding to the different drillcore samples, the drilling and sawing fluids as well as laboratory (PCR reaction+DNA extraction kit) associated contaminants. Sequence data have been deposited in GenBank ( with accession numbers FJ854696-FJ854749.

Phylogenetic and statistical analyses

Only high-quality partial sequences (700–800 bp) were retained for subsequent analyses. We discarded sequences of poor quality or potential chimeras. This yielded a total of 1578 sequences (Figure 2) that were used for phylogenetic and statistical analyses. Partial sequences were compared to those in databases by BLAST [44]. Multiple alignments were carried out using MUSCLE [45] and manually edited using the program ED from the MUST package [46]. Preliminary distance (neighbor-joining) trees allowed the identification of groups of highly similar sequences (>97% identity) or phylotypes. One representative clone from each phylotype was fully sequenced. Complete sequences were used to reconstruct a maximum likelihood phylogenetic tree with TREEFINDER [47], applying a general time reversible model of sequence evolution (GTR), and taking among-site rate variation into account by using an six-category discrete approximation of a Γ distribution (Figure 2). ML bootstrap proportions were inferred using 1000 replicates. Rarefaction curves were estimated out of the distance matrix from the different sequence alignments using the program DOTUR and the conventional species cut-off limit (members of a same OTU have >97% SSU rDNA sequence identity) [48].

Supporting Information

(0.07 MB PDF)

We thank the Institut de Physique du Globe de Paris, the Institut de Sciences de l'Univers, and the Geological Survey of Western Australia for supporting the PDP. This paper is published with the permission of the Executive Director of the Geological Survey of Western Australia.

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H.D.H K.T Treatise on Geochemistry Oxford Elsevier Pergamon 63 115 PedersenK 2000 Exploration of deep intraterrestrial microbial life: current perspectives. FEMS Microbiol Lett 185 9 16 NewmanDK BanfieldJF 2002 Geomicrobiology: how molecular-scale interactions underpin biogeochemical systems. Science 296 1071 1077 ParkesRJ WebsterG CraggBA WeightmanAJ NewberryCJ 2005 Deep sub-seafloor prokaryotes stimulated at interfaces over geological time. Nature 436 390 394 TeskeAP 2005 The deep subsurface biosphere is alive and well. Trends Microbiol 13 402 404 PedersenK ArlingerJ ErikssonS HallbeckA HallbeckL 2008 Numbers, biomass and cultivable diversity of microbial populations relate to depth and borehole-specific conditions in groundwater from depths of 4-450 m in Olkiluoto, Finland. Isme J 2 760 775 Van KranendonkMJ PhilippotP LepotK 2006 The Pilbara drilling project: c. 2.72 Ga Tumbiana Formation and c. 3.49 Ga Dresser Formation, Pilbara Craton, Western Australia. Western Australia Geological Survey, Record 2006/14, 25p: Western Australia Geological Survey. 25 p HayesJM 1994 Global methanotrophy at the Archean-Proterozoic transition. BengtsonS Early Life on Earth New York Columbia University Press 220 236 SchopfJW 2006 Fossil evidence of Archaean life. Philos Trans R Soc Lond B Biol Sci 361 869 885 OnstottTC MoserDP PfiffnerSM FredricksonJK BrockmanFJ 2003 Indigenous and contaminant microbes in ultradeep mines. Environ Microbiol 5 1168 1191 MoisslC OsmanS La DucMT DekasA BrodieE 2007 Molecular bacterial community analysis of clean rooms where spacecraft are assembled. FEMS Microbiol Ecol 61 509 521 LepotK BenzeraraK BrownGE PhilippotP 2008 Microbially influenced formation of 2,724-million-year-old stromatolites. Nat Geoscience 1 118 121 StapletonRD BrightNG SaylerGS 2000 Catabolic and genetic diversity of degradative bacteria from fuel-hydrocarbon contaminated aquifers. Microb Ecol 39 211 221 ShiT FredricksonJK BalkwillDL 2001 Biodegradation of polycyclic aromatic hydrocarbons by Sphingomonas strains isolated from the terrestrial subsurface. J Ind Microbiol Biotechnol 26 283 289 MiyakeD IchikiS TanabeM OdaT KurodaH 2007 Thiosulfate oxidation by a moderately thermophilic hydrogen-oxidizing bacterium, Hydrogenophilus thermoluteolus. Arch Microbiol 188 199 204 LavireC NormandP AlekhinaI BulatS PrieurD 2006 Presence of Hydrogenophilus thermoluteolus DNA in accretion ice in the subglacial Lake Vostok, Antarctica, assessed using rrs, cbb and hox. Environ Microbiol 8 2106 2114 KimuraH SugiharaM YamamotoH PatelBK KatoK 2005 Microbial community in a geothermal aquifer associated with the subsurface of the Great Artesian Basin, Australia. Extremophiles 9 407 414 FredricksonJK BalkwillDL DrakeGR RomineMF RingelbergDB 1995 Aromatic-degrading Sphingomonas isolates from the deep subsurface. 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Appl Environ Microbiol 70 4911 4920 KansoS GreeneAC PatelBK 2002 Bacillus subterraneus sp. nov., an iron- and manganese-reducing bacterium from a deep subsurface Australian thermal aquifer. Int J Syst Evol Microbiol 52 869 874 DornRI OberlanderTM 1981 Microbial origin of desert varnish. Science 213 1245 1247 CoxTL SlyLI 1997 Phylogenetic relationships and uncertain taxonomy of Pedomicrobium species. Int J Syst Bacteriol 47 377 380 BaxterNJ ScanlanJ De MarcoP WoodAP MurrellJC 2002 Duplicate copies of genes encoding methanesulfonate monooxygenase in Marinosulfonomonas methylotropha strain TR3 and detection of methanesulfonate utilizers in the environment. Appl Environ Microbiol 68 289 296 ErcoleC SulpiziiMP VeglioF BartoliniS ToroL 1997 Desulfurization of model aromatic molecules by Pedomicrobium sp. Fuel Processing Technology 52 191 197 LindahlT 1993 Instability and decay of the primary structure of DNA. Nature 362 709 715 CanoRJ BoruckiMK 1995 Revival and identification of bacterial spores in 25- to 40-million-year-old Dominican amber. Science 268 1060 1064 VreelandRH RosenzweigWD PowersDW 2000 Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal. Nature 407 897 900 FishSA ShepherdTJ McGenityTJ GrantWD 2002 Recovery of 16S ribosomal RNA gene fragments from ancient halite. Nature 417 432 436 InagakiF OkadaH TsapinAI NealsonKH 2005 Microbial survival: the paleome: a sedimentary genetic record of past microbial communities. Astrobiology 5 141 153 HebsgaardMB PhillipsMJ WillerslevE 2005 Geologically ancient DNA: fact or artefact? Trends Microbiol 13 212 220 DamsteJS CoolenMJ 2006 Fossil DNA in cretaceous black shales: myth or reality? Astrobiology 6 299 302 LambertDM RitchiePA MillarCD HollandB DrummondAJ 2002 Rates of evolution in ancient DNA from Adelie penguins. Science 295 2270 2273 ZhangG DongH XuZ ZhaoD ZhangC 2005 Microbial diversity in ultra-high-pressure rocks and fluids from the Chinese Continental Scientific Drilling Project in China. Appl Environ Microbiol 71 3213 3227 RogersSO TheraisnathanV MaLJ ZhaoY ZhangG 2004 Comparisons of protocols for decontamination of environmental ice samples for biological and molecular examinations. Appl Environ Microbiol 70 2540 2544 AltschulSF MaddenTL SchafferAA ZhangJ ZhangZ 1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25 3389 3402 EdgarRC 2004 MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32 1792 1797 PhilippeH 1993 MUST, a computer package of Management Utilities for Sequences and Trees. Nucleic Acids Res 21 5264 5272 JobbG von HaeselerA StrimmerK 2004 TREEFINDER: a powerful graphical analysis environment for molecular phylogenetics. BMC Evol Biol 4 18 SchlossPD HandelsmanJ 2005 Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71 1501 1506


Options On The Guitar

By Susan Tucker Guitar, Music Career, Songwriter, Songwriting No Comments on Options On The Guitar

Options On The Guitar

Guitarists often learn chords by reading “box” diagrams. If you’ve ever had a lesson in your life (or even if you haven’t) you’re probably familiar with these: essentially a drawing of the guitar

songwriter guitar

Dave Isaacs

neck held vertically, with six vertical lines representing the strings and horizontal lines representing the frets. Fingers are shown  by either dots or numbers on the notes you cover to create each chord.

Now, this is a useful tool….once you understand how to read these boxes, learning a new chord is as simple as looking at the picture. The problem is that many players never get beyond the concept of chords AS boxes: as fixed objects instead of a collection of notes.

A chord is any sound formed by more than two notes simultaneously.  Some on my young students like to grab random collections of notes and ask “is this a chord?” The answer is always yes, no matter how dissonant the sound might be…but the primary language of popular music is major and minor chords, and almost every other type of chord can be derived from this basic foundation.

Major Chords – Minor Chords

A quick definition. “Major” and “minor” are different qualities a chord can have, with two different sounds. (We’re using “quality” to mean a characteristic, as opposed to a judgment of good or bad). Every type of chord has a distinct quality and emotional association to the listener: to be very simplistic about it, major chords are bright and uplifting while minor chords are more dark and moody. Try playing a sequence of minor chords and end with a major…it sounds (and feels) like the sun breaking through the clouds.  (As an aside, one thing I love about great country music is how sad major chords can sound with the right melody, lyric, and delivery. Think of George Jones singing “He Stopped Loving Her Today”).

A little simple theory. Major and minor chords are made up of three notes, the first (root), third, and fifth of the corresponding scale; these three notes together form a triad. This is a simple enough idea to understand if you just start from the letter that names the chord, call that note 1, and count letters to find notes 3 and 5. So if we start from A and call that note 1, notes 3 and 5 would be C and E.

To work with this major and minor concept, strum a simple C chord followed by an A minor.  Listen to the different qualities of these two chords…the bright chime of the C, the major, and the darker sound of the A minor. Then reverse the order and listen to the emotional lift that accompanies the change from A minor to C.

Both the C and Am chords as we usually first learn to play them are 5-note chords, which means that some notes must be doubled. A simple C triad is C-E-G, while our 5-note C chord on the guitar is C-E-G-C-E.  That’s 1-3-5-1-3, for those who are keeping track. The A minor has a similar form: the triad is A-C-E, and the guitar chord is A-E-A-C-E or 1-5-1-3-5. Again, note that the numbers are always counted from the root, or note that names the chord: so C is note 1 of the C chord while A is note 1 of the A minor.

New Versions of Guitar Chords

Now here’s where we leave the box behind. You’re probably accustomed to playing all five notes of both these chords. Try holding the shape of either chord and playing just three strings at a time. You can extract three different versions of each chord this way: striking the three lower, three middle, or three upper strings. Notice how this brings out different notes, and adds an element of melody when you change chords or even just parts of the same chord.

Playing the same chords as arpeggios gives us even more options. To play an arpeggio, hold the chord form with your fretting hand and strike or pluck the strings one at a time. Notice how this also brings in a melodic element even without changing chords, and even more so when we put a series of chords together.

This is a large topic but a simple one. Try applying these ideas to other chords, bringing out different notes. As you change chords notice how melodies are created in this way:  now, instead of a series of “blocks” a chord progression becomes a series of simultaneous melodies. This is a great way to create signature parts that help define the song musically before the vocal even begins.

-Dave Isaacs-

To learn more about Dave  www.daveisaacs.com

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Production Associate-Embroider

This job is no longer active. View similar jobs.

POST DATE 8/31/2016
END DATE 11/19/2016

Things Remembered Fort Worth, TX

Things Remembered
Job Classification
Full Time
Company Ref #
Fort Worth, TX
Entry Level (0 - 2 years)
Job Type
AJE Ref #

The Production Associate represents Things Remembered to our customer. This position has primary responsibility for embroidery, engraving, customer satisfaction, generating sales, housekeeping, merchandising, POS operations and loss prevention, in adherence with all company/store standards.

*Demonstrates success in selling. Uses selling techniques such as add-on sales and describing engraving information to customers to enhance salesmanship and reach store and individual sales goals. This requires constant standing and walking to work with customers and show them merchandise.

*Is proficient in the basic skills of store operations: embroidery; engraving; performance of all register transactions; and equipment maintenance. This requires constant standing at the register and equipment as well as occasional squatting and kneeling.

*Participate in store activities of stocking, processing incoming and outgoing merchandise shipments. This requires constant standing and walking, occasional squatting and kneeling, frequently lifting & carrying up to 10 lbs and occasionally 11-30 lbs, as well as frequently climbing a 3-foot ladder working with arms overhead and occasionally using a 3-10-foot ladder to work with arms overhead.

(*Essential Functions)

Embroidery experience

Retail or customer service experience, preferably in a selling environment

Understands basic retail concepts: selling, visual merchandising, and loss prevention

Understands the importance of and is motivated by achieving sales goals and continually strives to increase sales

Demonstrates effective interpersonal skills and the ability to communicate verbally in a clear, audible, and grammatically correct manner

Ability to listen/read and accurately transcribe and verify customer message specifications onto engraved merchandise.

Relates to all customer segments and creates a good first impression

Manages multiple tasks and time effectively

Works as part of a team

Maintains a high energy level

Identifies the specific needs of the customer and suggests appropriate gift items.

Accepts suggestions and criticism in a positive manner and acts on them appropriately

Understands importance of teamwork in the store

Displays pride, self-confidence and a positive attitude

Is an active learner who takes responsibility for personal development

Has a strong work ethic and sense of personal responsibility

Is goal-oriented

Ability to work unsupervised

Displays flexibility to work a varied schedule to meet store needs
Things Remembered, Inc. is proud to be an Equal Opportunity/Affirmative Action employer.


Barack Obama

Barack Hussein Obama was the very first black president of the United States of America. His career as president started in 2009 and lasted 8 years, which equals 2 terms, ending with Trump’s victory in 2017.

Throughout his political career, he developed crucial international relations; for example, he put an end to war with Cuba which lasted around 50 years.

He possesses outstanding leadership qualities, he was loved by the people, kept promises that he had made during his campaign, and brought equality between races and balance to social differences.

Furthermore, he has a gorgeous family: a wife, whose name is Michelle and two daughters, who are called Natasha and Malia. He also has two beloved dogs.

Filipa, Patricia, Helena – FCE1Y M/W 18.50 (Martin)

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Current month ye@r day *


Monthly Message from the Superintendent

 April 2016

April is the time of year when all of the hard work and learning throughout the year appears to coalesce.  Our students continue to work hard connecting concepts and diving deep into engaging content.  They are able to apply their understanding of standards in meaningful ways to demonstrate their learning.

During a three-week window (May 2 – May 20), our students in third through sixth grade will participate in the state academic testing program, California Assessment of Student Performance and Progress (CAASPP).  CAASPP is intended to provide information that can be used to monitor our students’ progress and ensure that all students leave high school ready for college and career.  CAASPP includes computer adaptive tests in English language arts and mathematics (Smarter Balanced assessments), as well as paper-based tests for science.

The hard work and learning accomplished by our students throughout the school year have prepared them for the testing experience. The Smarter Balanced assessments are an academic check-up, designed to give teachers the feedback they need to inform instruction. The tests measure critical thinking, analytical writing, problem solving, and subject area knowledge, providing teachers with multiple sources of actionable information about student strengths and areas where students need additional support. Additional information about CAASPP can be found at this link.

Attendance at school on time, each day is an ongoing priority for every student.  It is even more important during our assessment window as it allows students the greatest opportunity to perform well.  Please ensure your children are well-rested and prepared for each day!

 We are excited about the remaining weeks of the school year and proud of our terrific students and staff!

Holly McClurg, Ph.D.

(858)755-9301 X3699


Seed Dance - Blindfolded Movement Journey with PrayerformanceART and special guest Adam Maalouf

Shamanique Experience Presents: The Seed Dance: Blindfolded Movement Journey w/ Live Music from Adam Maalouf THE SEED DANCE is an opportunity to EMBODY one’s most soulful INTENTIONS. Through blindfolded movement and an integrative dance wave, we grow our roots into the fertile darkness. In the exploration of our heartfelt expressions, we can nourish our seeds for the purpose of blossoming into our authenticity and deepen into the nature of our human experience. This event calls out to SPIRITUAL ENTHUSIASTS, NATURE LOVERS, ARTISTS, HEALERS, ENTREPRENEURS, VISIONARIES and anyone who is willing to be courageous towards living their dreams and vulnerable enough to make their mark in the world. Doors open and warm-up : 6:30 PrayerformanceART and Dance : 7:00-9:00 After dance: Tea and Reflections Wear comfortable clothes you can move in.

Doors and warm up at 6:30pm dancing begins at 7pm to 9:30pm

LIMITED TICKETS: (most likely will sell-out) $25 pre-purchase (plus service fee) $25-35 sliding scale at the door Pre-purchase tickets @

Seed Dance Facilitators:

Dominique Warfield (Asheville) is a catylyst for soulful transformation. She is the firestarter of the "Shamanaiuqe Experience" and has been touring around the country offering the Seed Dance. She is passionate about movement, music and spending time in the Appalachian waterfalls. Dominique aims to open the doors for authentic expression as a passage way for healing, empowerment and respect for our beautiful planet.

Adam Maalouf (New York) musical journey began at the age of 3, and after experiencing a wide range of musical training that included a classical education on the piano and cello, his eyes were opened to the infinite world of percussion. Adam's first experience with hand drumming was on the Arabic Derbekke Drum (otherwise known as the Doumbek) during his childhood visits to Koura, Lebanon. Since then, he has performed in many of the world's finest concert halls and museums, international music festivals, yoga and dance studios, public schools, and homeless shelters.

Lyca Mikhael (Asheville) is a Shape-Shifting Mage of Elemental Harmony and Lucid Movement ~ Founded in a wide library of Exploration, Evolution, and Expression. He is a genuine movement facilitator, drawing from deep roots, a vivid engagement with the present, and the flowing possibilities of perpetual arrival. Lyca is also a performer, spanning experimental dance through theatrical circus – working with his bare body and a wide library of tools like crystal spheres and enigmatic hoops. Lyca's Movement Arts Stoke Kindled Fires, Share Archetypal Stories, and Evoke Inner Light.


Luscious lemon sago recipe

After five attempts at making sago, I have finally found the answer. It still looks like frog’s eggs, but it’s a light and refreshing dessert.

Sago is sometimes sold as ‘seed tapioca’. The one  I used is made by McKenzie’s and they clarify the difference on the back of the packet:

A common alternative to Sago is Seed Tapioca. Sago and Tapioca are both starch extracts, sago from various southeast sago palms and tapioca is processed from the tubers of the cassava plant.

This explains my mixed results. Perhaps I should rename the recipe to Luscious Lemon Seed Tapioca? Naw, it doesn’t have the same ring to it.

You may like to try this recipe with limes instead. I like to eat lemon sago alone, but you can serve it with fresh or stewed fruit and cream or custard.

Luscious lemon sago recipe

1/2 cup of sago
2 cups of water
1 lemon, juiced
2 Tbsp honey
2 Tbsp golden syrup

  1. Wash the sago and soak for an hour in some water.
  2. Rinse off the water. Add 2 cups of water to a saucepan and bring to the boil.
  3. When boiling, add the sago and stir until quite transparent (this can take 15-30minutes).
  4. Add the lemon juice, golden syrup and sugar.
  5. Pour into a bowl or mould and chill.

Serves 4.


NASA's latest humanoid robot 'Valkyrie' may explore Mars (with video, photos)

Almas Meherally, Vancouver Sun

NASA engineers have created a 6.2 ft tall female-looking superhero robot 'Valkyrie' that could do tasks considered too dangerous for humans.

Valkyrie weighs 125 kgs, has 44 degrees of freedom (movement in its joints), a battery backpack and is capable of performing dynamic, dexterous and perception-intensive tasks in a variety of scenarios, according to its creators.

It has interchangeable arms and is covered in soft cloth that also doubles up as protection in case of a fall.

NASA Johnson Space Center hopes to see the robot on a future mission to Mars.

Valkyrie will be a part of the DARPA Robotics Challenge from December 20-21 that aims to create robots that can help people during natural and other kinds of disasters.

The DARPA challenge requires a robot to be compatible with environments engineered for people, use tools and be supervised by people who aren’t necessarily trained to operate robots.