text:
- Review
- Open Access
Cellulose degradation: a therapeutic strategy in the improved treatment of Acanthamoeba infections
- Sahreena Lakhundi1,
- Ruqaiyyah Siddiqui1 and
- Naveed Ahmed Khan1Email author
© Lakhundi et al.; licensee BioMed Central. 2015
- Received: 3 September 2014
- Accepted: 5 January 2015
- Published: 14 January 2015
Acanthamoeba is an opportunistic free-living amoeba that can cause blinding keratitis and fatal brain infection. Early diagnosis, followed by aggressive treatment is a pre-requisite in the successful treatment but even then the prognosis remains poor. A major drawback during the course of treatment is the ability of the amoeba to enclose itself within a shell (a process known as encystment), making it resistant to chemotherapeutic agents. As the cyst wall is partly made of cellulose, thus cellulose degradation offers a potential therapeutic strategy in the effective targeting of trophozoite encased within the cyst walls. Here, we present a comprehensive report on the structure of cellulose and cellulases, as well as known cellulose degradation mechanisms with an eye to target the Acanthamoeba cyst wall. The disruption of the cyst wall will make amoeba (concealed within) susceptible to chemotherapeutic agents, and at the very least inhibition of the excystment process will impede infection recurrence, as we bring these promising drug targets into focus so that they can be explored to their fullest.
- Acanthamoeba
- Cyst formation
- Cyst wall
- Cellulose
Protists are the largest, unicellular, non-photosynthetic eukaryotes that lack cell walls and include pathogens of both animals and humans. Pathogenic and opportunistic free living protist such as Acanthamoeba spp., Balamuthia mandrillaris and Naegleria fowleri occur worldwide and propagate independently in the environment. However upon accidental encounter with humans, they can produce disease [1-3]. Acanthamoeba species are now becoming increasingly important as human pathogens causing serious and life threatening infections. They are well known to cause blinding keratitis as well as granulomatous amoebic encephalitis [4]. The most distressing aspect of Acanthamoeba infections is their recurrence owing to the failure of therapy, attributed to the ability of the parasite to transform into a resistant cyst form [1-3]. Acanthamoeba undergoes two stages during its life cycle: a metabolically active "trophozoite" stage and a "cyst" stage with minimal metabolic activity. The trophozoite aggressively feeds on human cells as well as grazes upon bacteria, yeast and/or small organic particles and multiplies enthusiastically. Exposure to harsh environmental conditions i.e., in the face of famine, extremes of pH, temperature, osmolarity and the presence of antimicrobial drugs, results in cellular differentiation into a resistant cyst form [5]. During encystment, the trophozoite enfolds itself into a circular structure surrounded by a cyst wall with minimal metabolic activity. The transformation of the trophozoite from a metabolically active form to an inactive form can impede successful treatment. When encysted, the trophozoite encloses itself within a shell that makes it resistant to physiological concentrations of a number of physical, chemical and radiological conditions and chemotherapeutic agents [6,7]. It is envisaged that degradation of the shell (cyst wall) will allow effective killing of trophozoite within. The cyst wall consists of an ectocyst (laminar, fibrous outer layer) and an endocyst (fine, fibriler inner layer) [5,8,9]. Both layers are separated by space except at opercula in the centre of ostioles which during excystation serves as the exit point for trophozoite. The mature cyst therefore consists of a double-walled structure with the wall serving as a protective barrier to facilitate parasite survival of hostile conditions. As the cyst wall offers resistance to chemotherapeutic agents, it may leave amoeba viable to re-establish infection following antimicrobial chemotherapy [6]. Cyst walls remain intact even with the treatment of SDS plus boiling, EDTA, enzymes (including papain, DNase, RNase, amyloglucosidase, proteinase K) and DTT [10]. Resilient nature of cyst walls suggest that they are composed, at least in part, of carbohydrates possibly polysaccharides. This is in agreement with the fact that the carbohydrate analysis of cysts walls of A. castellanii using GC/MS showed that they contain 44.4% glucose [10]. The linkage analysis confirmed the presence of 4-linked glucopyranose (22.2%) in the cyst walls of A. castellanii which is suggestive of cellulose and is consistent with previous findings [11,12]. Dudley et. al., [13] observed the inhibition of Acanthamoeba encystment in the presence of 2,6-dichlorobenzonitrile, an inhibitor of cellulose synthesis. Furthermore, the enzymes for the synthesis and breakdown of cellulose have been identified in A. castellanii, indicating the presence of cellulose in the cyst walls of this organism [14]. Hence the sturdy nature of Acanthamoeba cysts is attributed, in part, to cellulose and suggests that cellulose could serve as a potential target. Previous work has described cellulose synthesis in Acanthamoeba [5,12,14-16]. In agreement with recent studies [17,18], here it is proposed that cellulose degradation offers a potential therapeutic strategy in effective targeting of trophozoite encased within the cyst walls. Here, we present a comprehensive report on the structure of cellulose and cellulases, as well as known cellulose degradation mechanisms with an eye to target Acanthamoeba cyst wall. It is envisaged that the disruption of cyst wall will make amoeba (concealed within) susceptible to chemotherapeutic agents, and at the very least inhibition of excystment process would impede infection recurrence and undoubtedly be of potential value in therapy.
What is cellulose?
Cellulose, although it is said to have a crystalline structure, in nature these fibers are not purely crystalline. In the physical world, cellulose fibers range from purely crystalline to purely amorphous forms [20,24]. Depending on the origin, the degree of cellulose crystallinity can vary from 0% of that of amorphous, acid swollen cellulose, to 70% of that from cotton, to nearly 100% of that from Valonia macrophysa [25,26]. The organization of individual microfibrils in crystalline cellulose is such that the component molecules are packed tightly enough to prevent penetration by enzymes. In addition, cellulose also contain various irregularities such as twists or voids, surface micropores, large pits and capillaries etc., increasing the total surface area much larger than that of an ideally smooth fiber of the same dimensions [20,27-29]. Hence the net effect of this heterogeneity is that the fibers are partially hydrated when in aqueous medium, allowing the invasion of comparatively larger molecules like cellulases. Overall, cellulose imparts high tensile strength to the wall it is contained in, serving as a structural component. It is an extracellular polysaccharide and is a part of the cell wall in plants, algae, bacteria, slime mold Dictyostelium and other protists such as Acanthamoeba cyst wall [30-33].
What are cellulases?
Endoglucanase activity
Exoglucanase activity (also called cellodextrinase or cellobiohydrolase)
β – glucosidases activity
Cellulose binding domain (CBD)
Catalytic domain (CD)
The longest domain within cellulases corresponds to catalytic cores. Studies of gene deletion, proteolytic truncation etc., show that they behave as independent entities conferred with catalytic activity and defined specificity towards soluble model substrates [41]. Although the CD of cellulases exhibit considerable diversity, they have been grouped into glycoside hydrolase families based on their amino acid sequence similarities. Glycoside hydrolases are a group of enzymes which hydrolyze the glycosidic bond between carbohydrates or between a carbohydrate and a non-carbohydrate moiety [20,55]. The enzymes in the same family contain similar basic fold based on the idea that a direct relationship exists between amino acid sequence and the folding of the protein. Based on this classification, enzymes having different substrate specificities are sometimes found in the same family; for example, family 5 contains cellulase, xylanases and mannanases, indicating an evolutionary divergence to acquire new specificities. On the other hand, enzymes with same substrate specificity are found in different families; for example, cellulases are found in 11 (5, 6, 7, 8, 9, 12, 44, 45, 48, 61 and 74) different families. This classification which contains more than 5000 glycoside hydrolases are grouped into 130 families. The classification of glycoside hydrolase into structurally determined families provides valuable insights into the structural features of the enzymes, which are more informative than the substrate specificity which was the basis of old IUBMB classification. The 3-D structure of one member of the family can be used to infer the structure of other members of the same family. This classification also defines the domains of the enzymes and thus resolves the contradiction about substrate specificity for multifunctional enzymes [20]. It also sheds light on the evolution of the glycoside hydrolases; as for example, some families are deeply rooted evolutionarily, such as family 9, which contains cellulases of fungi, bacteria, animals and plants. However this is in contrast to family 7 which contains hydrolases of fungal origin only and family 8 which contains only bacterial hydrolases. In addition, cellulases from several families and hence from several different folds are found in the same organism; for example, Cellulomonas thermocellum contain endoglucanase and exoglucanase from families 5, 8, 9 and 48. Cellulases are hence a complex group of enzymes that seems to have evolved through concurrence from a repertoire of basic folds. In addition, the extensive diversity within the cellulase families reflects the heterogeneity of cellulose and associated polysaccharide within plant material and a variety of environment where hydrolysis takes place. Proteolytic truncation of CBDs led to the determination of the 3D structures of a variety of CDs by X-ray crystallography. The CD of Trichoderma reesei cellobiohydrolase II (CBH II), belonging to glycosyl hydrolase family 6, is a large α/β protein with 5 α-helices and 7 β-strands. The active site is an enclosed tunnel located at the C-terminal end of a β-barrel through which the cellulose chain threads [56]. This structure was confirmed in endoglucanase II from Thermomonospora fusca, another family 6 member [57]. The 3D structures of catalytic cores of both these enzymes are very similar. However, closer examination reveals a major difference in the degree of active site accessibility to the substrate corresponding to the major difference in their endo- and exo- mode of action [58]. In endoglucanase, the substrate tend to lie in an open cleft which can accommodate cellulose anywhere along the chain, whereas exoglucanase forms an enclosed tunnel in order to allow cellulose chains to be threaded releasing cellobiose or cellooligosaccharides. The hydrolysis of β 1,4-glycosidic bond proceeds via general acid–base catalysis requiring a proton donor and a nucleophile or a base [59,60]. The hydrolysis product can either result in overall retention or inversion of the configuration of the carbon at anomeric position. The stereochemical course of hydrolysis has been determined for several glucanases belonging to different families [61-66]. For example, members of the family 5, 7 and 11 proceeds via retention of the anomeric configuration, whereas enzymes of family 6 and 9 proceeds via inversion mechanism. Generally the overall 3D structure and stereospecificity of hydrolysis are conserved within the family.
Cellulose degradation mechanisms
There are at least four different known strategies for cellulose degradation as described below.
Non-Complexed cellulolysis
Complexed cellulolysis
Presence of correct ratio between components of cellulosomal machinery optimizes synergism.
Optimum spacing and organization of individual components maximizes synergy by avoiding non-productive binding.
Competition between different components is limited due to the binding of the whole complex to a single site.
The presence of enzymes with different specificity in one complex avoids the halt in the process on depletion of one structural type of cellulose.
Cellulolysis without processive cellulases
The third strategy appears to be utilized by at least two very efficient cellulose degraders: Cytophaga hutchinsonii, an aerobic soil cellulolytic organism and Fibrobacter succinogenes, an anaerobic ruminal bacterium [34,35,88]. These organisms do not seem to encode for any processive cellulases. Up until recently, the presence of processive cellulase was considered mandatory for effective degradation of crystalline cellulose. Examination of C. hutchinsonii genome sequence reveals the presence of clusters of genes responsible for degradation of cellulose [38]. In addition to genes encoding probable β endoglucanases, there are genes encoding possible β glucosidases but no recognizable exoglucanases/cellobiohydrolases. In addition, most cellulases of C. hutchinsonii do not contain CBD indicating the absence of any processive cellulases [34,98]. As described above, cellulases generally consist of a CD, a CBD and the linker, joining the two [53,99]. Since to date these features were considered mandatory for the efficient degradation of cellulose by other cellulose degraders, C. hutchinsonii may therefore have a novel mechanism for cellulose degradation. The genome sequence of the major cellulolytic rumen bacterium F. succinogenes has been determined [34]. It does not seem to possess any processive cellulases nor does it seem to encode for dockerin domains or a scaffoldin gene, as does other anaerobes [100,101]. Furthermore, none of the cellulases from these organisms have much activity on crystalline cellulose [35,38,102]. However both these organisms grow very rapidly on cellulose. They have a unique requirement of physical association with their carbon source and are only able to grow when tightly bound to cellulose. In addition, none of these organisms secrete free cellulases or produce cellulosomes. Hence there appears to be a novel strategy for cellulose utilization by these organisms. One possible mechanism for cellulose digestion by these bacteria might be the one used by starch degrading bacterium, Bacteroides thetaiotaomicron [34]. In this model, cellulose is bound to a protein complex present in the outer membrane and the individual molecules are transported into the periplasmic space, where they are degraded by cellulases. In this model, individual cellulose molecules would be readily degraded by endoglucanases and hence excludes the requirement for processive cellulases. This hypothesis also fulfills the unique requirement of these bacteria to be in direct contact with their insoluble substrate for efficient digestion [34,35,38].
Cellulolysis via non-hydrolytic enzymes
Lastly, some non-hydrolytic enzymes i.e., oxidative enzymes have also been reported to be associated with cellulose degradation [103]. This system is most thoroughly studied in white-rot fungi, however some cellulolytic bacteria have also been discovered which tend to degrade cellulose via oxidative enzyme strategy [97,103,104]. In addition to hydrolytic enzymes, occasional non-hydrolytic enzymes have also been cited in the literature to be involved in cellulose degradation [103,104]. These include oxidative enzymes such as cellobiose:quinone oxidoreductase (CBQ), cellobiose dehydrogenase and cellobiose oxidase (CBO) [103,105-107]. These enzymes oxidize cellobiose and/or higher cellodextrin reducing ends to their corresponding lactones which can then be used by organisms as their source of carbon [97,104,108]. After the discovery of oxidative enzymes as potential candidate involved in cellulose degradation [103], research has mostly been focused on enzymes from white rot fungi. To date, there is little knowledge about oxidative systems in bacteria [97,104]. The possible role played by these oxidizing enzymes in cellulose degradation may be to alleviate the inhibitory effect of cellobiose on cellobiohydrolase action, to regulate the synthesis of enzymes involved in cellulose degradation and/or in the metabolism of cellobiose itself [97]. In addition since these enzymes are most commonly found in wood rotting fungi, it is suggested that oxidases might also generate H2O2 required by lignin-degrading enzymes like peroxidases and laccases for the degradation of lignin in natural environment.
Synergy and competition among cellulose degrading system
Synergism between endoglucanase and exoglucanase, called endo-exo synergy
Synergism between exoglucanases, called exo-exo synergy
Synergism between exoglucanase and β – glucosidase to remove cellobiose that inhibits exoglucanase
Synergism between catalytic and carbohydrate binding domains
Fujita et al., [109] reported the synergistic hydrolysis of amorphous cellulose by a yeast strain co-displaying endoglucanase II (EGII) and cellobiohydrolase II (CBHII) from Trichoderma reesei and Aspergillus aculeatus β-glucosidase 1 (BGL1). They observed higher hydrolytic activity by the strain co-displaying EGII and CBHII (1.3 mM reducing sugar were released in 60 hours) on amorphous cellulose than the strain displaying only EGII (0.5 mM reducing sugars were released in 60 hours) with the main hydrolysis product been cellobiose. The co-display of BGL1 along with EGII and CBHII resulted in direct production of ethanol from amorphous cellulose. Ethanol was not produced from amorphous cellulose in the presence of only EGII and CBHII. Zhou and Ingram [110] observed synergism between two endoglucanases CelY and CelZ from Erwinia chrysanthemi. They observed about 1.8 fold synergy when the enzymes were used in combination. The synergy was due to the difference in substrate preference. CelY hydrolyzed CMC to fragments averaging 10.7 glucosyl units but was unable to hydrolyze cellotetraose and cellopentaose. On the other hand CelZ readily hydrolyzed soluble cellooligosaccharides and amorphous cellulose to produce cellobiose and cellotriose as major end product. Hydrolysis of CMC by CelZ resulted in fragments averaging 3.6 glucosyl units. In combination both enzymes hydrolyzed CMC to fragments averaging 2.3 glucosyl units. Synergy was also observed after the sequential addition of CelY and CelZ (after heat inactivation of CelY) showing that synergy does not require the simultaneous presence of both the enzymes. However no synergy was observed when CelZ was used as the first enzyme, hence showing that only CelY can act independently to modify the substrate to make it more accessible to CelZ. By conventional definition, exocellulases releases cellobiose from non-reducing ends of cellulose chain, which does not explain exo-exo synergy and would hence compete for limited number of hydrolysis sites, instead of cooperating to give synergistic hydrolysis. Enzymatic hydrolysis would be insufficient if the enzymes only act at non-reducing ends, since half the chain ends would be unused. Hence this traditional model was questioned followed by the report describing two exoglucanases from Aspergillus aculeatus, one of which attacks from reducing end and the other from non-reducing end of cellulose chain. Similarly two different classes of cellobiohydrolases in Trichoderma reesei, CBHI (makes up 60% of total cellulolyticproteins) acting from reducing end of the chain and CBHII (makes up 20% of total cellulolytic proteins) acting from non-reducing end can achieve complete solubilisation, although slow, of cellulose without the help of endoglucanases [40]. Hence these data suggests that the observed exo-exo synergy could be due to the interaction between non-reducing end attacking and reducing end attacking exocellulases.
Most cellulases are composed of CD and CBD that function independently but act synergistically in the disruption and hydrolysis of cellulose fibres. The CBD makes the substrate more accessible to hydrolytic domain by bringing the catalytic module in close proximity to boost hydrolysis [20,45,48]. It is also considered to play a role in sloughing off the cellulose fragments from the surface of cellulose by splitting of the cross linkages. It is also worth mentioning that cellulose degraders always seem to produce multiple enzymes of each class. As an example Trichoderma reesei cellulase system consists of two exoglucanases (CBHI and CBHII), five endoglucanases (EGI, EGII, EGIII, EGIV and EGV) and two β-glucosidases (BGLI and BGLII) [71]. Humicola insolens produces at least seven cellulases including two cellobiohydrolases (CBHI and CBHII) and five endoglucanases (EGI, EGII, EGIII, EGV and EGVI) [79]. Thermobifida fusca contains three endoglucanases (E1, E2 and E5), two exoglucanases (E3 and E6) and a cellulase with both endo-exo activity (E4) [20,85]. The secretion of multiple cellulases of same class could be due to the heterogenous nature of their substrate. As cellulose structure varies from being purely crystalline to purely amorphous with all degrees of order in between, hence some of these enzymes are more effective towards one form of cellulose while others are more effective towards other forms. In addition, it also indicates that although each individual β 1,4-glucosidic bond is chemically identical, the complex nature of the substrate and the environment in which they are present shows that they are not in identical context. Hence therefore one might expect to observe synergy between cellulases of same class as well as of different classes. The combination of these enzymes thus acts synergistically to hydrolyse cellulose. In addition to cellulase these organisms may also possess enzymes to degrade other polysaccharides associated with cellulose such as hemicelluloses, presumably because their breakdown is required to gain access to cellulose fibres [38].
With regard to synergy between cellulases, synergy is observed in some cases but not in other. Anderson et al., [111] reported synergistic effect by a mixture of cel45A, cel6A (an endoglucanse and an exoglucanase respectively from Humicola insolens) and β-glucosidase from Penicillium brasilianum on amorphous cellulose. However on crystalline cellulose, these enzymes seem to rather inhibit each other owing to the competition for binding sites on cellulose. Whereas some other studies [112,113] with different enzymes showed synergistic effect on crystalline cellulose but not on amorphous cellulose.
Cellulolytic protists
Most cellulolytic protists are anaerobes, found in the rumen responsible for the degradation of plant material [36]. Cellulolytic flagellated protists are the major cellulose degraders present in the hind gut of lower termites. In addition, the hind gut of wood eating cockroaches harbor many symbiotic protists responsible for the digestion of cellulose rich diets [36,114,115]. The protists tend to endocytose cellulose particles into their food vacuoles whereby, cellulases produced by protists degrade cellulose [116]. However cellulases from protists have hardly been characterized at molecular level due to the difficulty in culturing the anaerobic gut protist community. So far only a limited number of species have been cultured axenically although culture independent techniques have led some advancement in identifying and characterizing cellulases from gut protists [117,118]. Using PCR, fifteen full-length cDNA clones were isolated and sequenced from the protist community of termite, Reticulitermes sepratus. They were found to be identical to family 45 cellulases and were originated from hypermastigote protists. Unlike other bacterial and fungal cellulases, the cellulolytic enzymes from protist consist of catalytic domain only [119]. Ancillary domains like cellulose or CBD were not present in these enzymes. The ingested cellulosic material is crunched and ground by the host increasing the surface of cellulose [120]. The cellulosic material is then selectively endocytosed into the food vacuole where it is degraded further by cellulases [119,121]. As mentioned earlier, the role of CBD is to anchor the enzyme to its substrate thereby increasing the concentration of enzyme at the surface of the polysaccharide. High enzyme concentration around the substrate in the food vacuole together with increased surface area of the ground and crunched cellulose may compensate for the lack of CBD in protist cellulases.
Coptotermes formasanus, a wood-feeding termite harbours three symbiotic parabasalian flagelates including Spirotrichonympha leidyi, Holomastigotoides mirabile and Pseudotrichonympha grassii [119,122]. These flagellates play an essential role in the digestion of cellulose rich diet. P. grassii is however found to utilize high molecular weight cellulose particles whereas; H. mirabile and S. leidyi tend to degrade low molecular weight cellooligosaccharides [119,123]. Nakashima et al., [122] isolated a novel cellulase gene, PgCBH-homos from the flagellated protist P. grassii. On the basis of amino acid sequence, PgCBH-homos is found to be similar to GHF 7 (Glycosyl hydrolase family 7) members that mainly consists of fungal cellulases. However clones similar to GHF7 have been detected in Dictyostelium discoideum, the social amoebae of the cellular slime mold (Dictyostelium discoideum cDNA Project at [124]. On the basis of their tertiary structures and the catalytic mode of action, GHF 7 members are subdivided into two groups: endoglucanases and cellobiohydrolases. PgCBH-homo however has shown to consist of regions similar to Cel7A from Trichoderma reesei, a processive cellobiohydrolase forming a tunnel shaped active site [122]. Hence it was predicted to have similar 3D structure to Cel7A and probably functions as a processive cellulase active against crystalline cellulose. The enzyme was also found to contain signal sequence at N-terminal corresponding to the fact that the enzymes could be secreted into the food vacuole after endocytosis of wood particles. Another cellulase gene coding for an endoglucanase CFP-EGI, from symbiotic protist Spirotrichonympha leidyi present in the hind gut of lower termite Coptotermes formasanus was cloned, expressed and characterized [119]. It's a 33.6 kDa protein and shows sequence similarity with members of GHF 5, a large and growing family of glycosyl hydrolases comprising of endoglucanases, xylanases, mannanases and 1,3-exoglucanases of both aerobic and anaerobic origins. In addition hydrolases of GHF 5 are very diverse and belongs to both prokaryotes and eukaryotes including bacteria, fungi, nematodes, protists and insects. The heterologous CFP-EGI was found to have an optimum pH of 5.8-6.0 and an optimum temperature of 70°C. In 2010, Todoka et. al., heterologously expressed and characterized a cellulase, RsSymEG, from a symbiotic protist of the lower termite, Reticulitermes speratus. The amino acid sequence of RsSymEG indicates that it is a GHF 7 cellulase. The lack of insertion sequence responsible for the formation of a tunnel shaped active site conserved among GHF7 CBHs, indicate that RsSymEG is an endoglucanase similar to that of Cel7B, an endoglucanase from Trichoderma reesei. However unlike Cel7B, no CBD was detected in RsSymEG and the optimum temperature and pH was found to be 45°C and 6.5 respectively. In addition chromatographic analysis revealed that the preferred substrate for RsSymEG is cellodextrin which is broken down into cellobiose and glucose. Using PCR, 11 different cellulases from symbiotic protists of four different termites were cloned in Saccharomyces cerevisiae [125]. The cellulases belong to 3 different glycosyl hydrolase families i.e., GHF 5, 7 & 45 and were found to be more efficient than EG I, a major endoglucanase from Trichoderma reesei, in degrading carboxymethyl cellulose. The presence of cellulases from a variety of different glycosyl hydrolase family corresponds to the fact that various cellulases act in synergism to bring efficient hydrolysis of polymeric cellulose [116]. Unlike anaerobic rumen protists that degrade cellulose, Dictyostelium discoideum, the amoebae of the cellular slime mold under harsh conditions transforms into a microcyst, containing cellulose rich cell wall [126-129]. Their cell wall mainly consists of four layers: 1, 2a, 2b and 3. Middle layers i.e., layer 2a and 2b are mainly composed of cellulose fibrils and layer 3 i.e., the inner most layer is at least made up of two components: cellulose and proteins [129,130]. As mentioned earlier the cellulose in their cell wall serves as the structural component and help survive hostile conditions. The inability of Dictyostelium mutants to synthesize cellulose results in the failure to form viable spores [33]. The spores remain dormant until the conditions are favourable which then stimulates spore germination. The germination is characterized by the activation of spore, budding of the cell wall with swelling of the entire spore and finally, lysis of the cell wall with simultaneous emergence of an amoeba [131]. Cellulolytic activity in D. discoideum, first observed by Rosness [127], plays a significant role in the germination process by causing lysis of the cell wall, hence allowing the emergence of the viable amoeba. Rosness observed the degradation of acid swollen cellulose to glucose by extracts from D. discoideum at the sorocarp stage of development. The cellulase activity was found to increase rapidly during aging of sorocarp and therefore the cellulases are thought to play a central role in the germination process of this organism. Latter Jones et al. [132], partially characterized two cellulolytic enzymes in this organism released during swelling stage of germination. During this stage the layers 1, 2a and 2b are ruptured and this hence suggests that cellulases during this stage act to degrade layers 2a and 2b celluloses and eventually diffuse into the extracellular medium after the rupture of the cell wall.
With the completion of the Acanthamoeba genome, enzymes for the synthesis and breakdown of cellulose have been identified in A. castellanii which are likely to participate in the morphogenesis in this group of organism [133,134]. Based on sequence data, cellulases of A. castellanii appear to be similar to bacterial proteins and belong to the glycosyl hydrolase family 5, while that of D. discoideum belong to glycosyl hydrolase family 9. Cellulases of GH family 5 are mainly endoglucanase which randomly cut cellulose chain in the middle releasing short chain oligosaccharides like cellodextrins. However, a complete identification, characterization, and localization of enzymes in Acanthamoeba with β-glucanase activity, which are likely necessary in morphogenetic events as well as controlled hydrolysis of the cyst wall remain unclear. Based on the known enzymes involved in various cellulose degradation systems, future studies should be carried out to specifically address the following question.
What is the specific structure of cellulose in Acanthamoeba? how are cellulose fibers in Acanthamoeba organized, ranging from crystalline to amorphous forms? and what is the specific combination of cellulases that can disrupt Acanthamoeba cyst wall integrity. These are important questions as in patient tissues, Acanthamoeba form cysts and are resistant to chemotherapy, leading to recurrence of infection after treatment. It is envisaged that the disruption of cyst wall will make amoeba (concealed within) susceptible to available chemotherapeutic agents. 'Associative' therapy should therefore act in conjunction and thus may augment the potency of other compounds, resulting in the improved treatment with reduced infection recurrence. Also cellulose is a structural component limited to some bacteria, protists and higher plants. Consequently, a specific cellulose degrader (i.e., cellulase) should ideally have no and/or minimal effect on non-target (human) cells. Studies should therefore focus on testing the use of these molecules/compounds in vivo which may aid in the improved therapy against Acanthamoeba infections. These are important questions as in patient tissues, Acanthamoeba form cysts and are resistant to chemotherapy, leading to recurrence of infection after treatment. In addition, a thorough understanding of cellulose degradation mechanisms and/or identification of components that can interfere with this process would undoubtedly be of potential value in therapy.
The authors would like to stress the fact that the work presented in this review is the results of the dedication and insights of many scientists who contributed considerably to our understanding of this fascinating organism. The work is supported by the Aga Khan University.
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Fire Poi Triple 50mm Oval 40cm Chain Double Leather
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text:
Moderator: Liquidcamphor
On a recent morning, MTA officials led a walking and stair-climbing tour to showcase progress on the unfinished, cavern-like terminal with ceilings as high as six stories. Dozens of high-speed escalators are being built to lead down to a 350,000-square-foot (32,516-square-meter) LIRR concourse with marble already laid on its walls and space reserved for retail shops and dining areas.
It will be the hub for 8 miles (13 kilometers) of new tunnels blasted and drilled out from 400 million-year-old bedrock, winding their way under Park Avenue and the East River and on to Queens and Long Island.
It is all focused on alleviating some of the pain for about 150,000 LIRR commuters who must somehow get to and from Grand Central every day. Currently, they have to joust, push and elbow their way to platforms at crowded Penn Station, which also accommodates Amtrak and New Jersey Transit trains, adding up to 40 minutes to their daily commutes.
Lieber says he is focused on hitting the Dec. 31, 2022, deadline.
Herring wrote:Project to be done by 2022
I'll be retired by then.
SRich wrote:Even thru a VPN i can't acces the link can you post a fair quote ?
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text:
A Lake Shore Drive address has a certain cachet in Chicago, and the new 500 Lake Shore Drive apartment tower aims to live up to that reputation.
The doorman wears white gloves. The spacious, two-story glass, wood and stone lobby soars in the manner of an upscale hotel, unlike some peers with more perfunctory entrances. The impressive array of amenities and services includes on-site pet care.
"Everything in this building is meant to make a renter's life easier," said Curt Bailey, president of Related Midwest, developer of the property.
Pets, for example, are welcome throughout the 500-unit building, and the open policy has lured some of the early residents who began moving into the studio to two-bedroom units on floors 3 to 29 in mid-May. Leasing for the remainder of the 47 floors begins in the fall.
Pet care is the primary concern, "by a mile," of apartment dwellers, Bailey said.
The solution is the addition of a full-service pet-care center as part of the 2,500 square feet of first-floor retail space. Dog City Chicago offers residents dog walking, grooming, training, overnight care and veterinary services as well as cat-sitting.
In addition to pet lovers and young urban professionals, Bailey expects "empty nesters and suburban homeowners wanting a second home downtown to be tenants.
"We think it is a big untapped market," he said.
The building, across from the busy Grand Avenue offramp from Lake Shore Drive, is part of a high-profile neighborhood. Lake Point Tower, Olive Park and Lake Michigan are across the street. Navy Pier, one of the city's top tourist attractions, is a short walk away.
The apartments range from convertible units from 550 square feet to three-bedroom penthouses of up to 2,800 square feet on the top four floors. Model apartments are sleekly contemporary, with floor-to-ceiling windows, wide-plank flooring and 9-foot ceilings. Most of the units have some water view, and those with east- and south-facing windows enjoy a bird's-eye view of the Navy Pier fireworks.
A 1,200-square-foot, two-bedroom, two-bath model on the northeast corner of the building shows off lake views than can be enjoyed from the kitchen that opens to the main living area.
The kitchen, equipped with Italian cabinetry, quartz countertops, white marble mosaic backsplash and built-in stainless steel appliances, is separated from the living area by an island with a countertop gas range. The stainless steel hood functions as a vent and design element.
The microwave is discreetly tucked inside kitchen cabinetry, and installation of the in-unit washer and dryer — they are standard in all apartments — is similarly unobtrusive.
One of the few plans without any glimpse of the lake is a convertible apartment that has its own charms. It faces west to the city's lights and has little wasted space, packing the available square footage into a side-by-side living area and sleeping/home office area. The plan has a natural division of public and private spaces while managing the feeling of elbow room.
There has been no stinting on the amenities in the 12th-floor 500 Club, anchored by a fitness center overlooking Olive Park and Lake Michigan. Residents can play in a game room with walls of reclaimed wood or avail themselves of the movie screening room, tech center, lounge and coffee bar, outdoor pool and deck with cabanas, fire pit and grills. A charming library is tucked between more active, screen-oriented spaces.
Among the services available in the smoke-free building is a 24-hour concierge, an on-site personal assistant to help with move-in, and an on-call technology service to hook up and set up home entertainment and electronics. The building is equipped with a high-tech elevator system designed to whisk tenants efficiently during those busy morning and evening hours.
The site has a distributed antenna system for enhanced in-building cellphone reception.
Transportation is no problem, as the 11-story garage includes private and public parking, a bike share program and an on-site Zip Car station with a Toyota Prius, Audi and BMW available, Bailey said.
500 Lake Shore Drive
Chicago, 60611; 312-850-0500; 500lakeshore.com
500 studio, convertible and one- to three-bedroom residences in the Streeterville neighborhood.
Apartments: Prices based upon availability and subject to change: studios, 600 sq. ft., from $1,795; convertible, one-bath, 550 to 750 sq. ft., from $1,995; one-bedroom, one-bath, 775 to 800 sq. ft., from $2,215; two-bedroom, two-bath, 1,150 to 1,550 sq. ft., from $3,695; three-bedroom, three-bath, 2,000 to 2,800 sq. ft., from $8,995.
Lease terms: 1-year lease. One-time application fee, $50 per applicant, and one-time administration fee, $450.
Renter's insurance: Recommended
Utilities: Tenant is responsible for utility and service amenity fee, $85 to $195, depending on size of apartment, includes heat, air conditioning, water, gas, Internet and cable. Electricity is individually metered.
Parking: Indoor reserved parking, $375 per month; unreserved parking, $275.
Pets: One-time dog fee, limit two dogs, $695, includes one-year Dog City membership. Breed restrictions, 75-pound weight limit. One-time cat fee, $200, limit two cats, includes one-year Dog City membership.
Smoking policy: Smoke-free
Amenities: Snaidero cabinetry, quartz countertops, built-in stainless steel kitchen appliances and hood. Natural stone flooring, walnut vanities in baths. Nine-foot ceilings, wide-plank flooring, fitness center, game room, library, screening room, coffee bar, tech center, and rooftop terrace with pool, hot tub, cabanas, fire pit and grill. Around-the-clock doorman and concierge services. Distributed antenna system to enhance cellphone service. Parking garage with reserved and public parking, and bike share and car share station. Building has applied for silver Leadership in Energy and Environmental Design certification.


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Olean and Salamanca City Court Judge William J. Gabler, Sr., announced…that he is a candidate for the office of Cattaraugus County Judge that will be filled this fall due to the mandatory retirement of Judge Michael L. Nenno at the end of this year. Judge Gabler has been endorsed by the Conservative and Democratic Parties.
Judge Gabler retired from his position as Cattaraugus County Family Court Support Magistrate at the end of 2013, where he served for 28 years. He has continued to serve as the Salamanca City Court Judge, a position he has held for the past 25 years; and, was appointed to the part-time Judge position in the Olean City Court in January after serving in that Court the past year on assignment from his post as Salamanca City Court Judge filling the void left by Judge William H. Mountain's retirement. In addition, he represents the Salamanca Industrial Development Agency and the Allegany County Department of Social Services' Support Collection Unit.
In making the announcement, Judge Gabler said, "This decision comes after a great deal of thought, soul searching and prayer. My decision was made more difficult by the political challenge to the designating petitions and the removal of my name from the Democratic line on this year's ballot. But, I believe it is more important for the voters of Cattaraugus County to have the opportunity to elect their next County Judge rather than our Judge being selected by a handful of political operatives in Little Valley. The nature of the work done by our County Courts is serious and sensitive and demands a judge with tested judicial experience. Just this year our County Court has heard a murder trial and other serious felony cases and our Family Court deals on a daily basis with intimate and emotional issues such as the custody and visitation of children, abuse and neglect of children and juvenile delinquency. Our County Courts are no place for on-the-job training."
He added, "I am 64 years old. I have been on the bench, hearing and deciding tens of thousands of cases in the New York State Family Court, Salamanca City Court and Olean City Court for nearly 31 years – essentially half of my life. Over those three decades I have been appointed, reappointed or elected 16 times. Sixteen times my record has been thoroughly vetted, approved and I have been continued in office – and my appointments have been made by Republican and Democratic administrations alike. I respectfully submit this stands as a testament to the quality of my judicial work, my work ethic and my judicial temperament. The voters of Cattaraugus County will not have to guess at the kind of County Judge I will be, they have only to look at my long judicial record for the answer."
In keeping with his record of non-partisanship and reflecting his history of bi-partisan support, Judge Gabler will be reaching out for support from voters in all political parties and independents alike.


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Send your friends a fall greeting wrapped up beautifully in this Tastes of Fall Gift Basket! Tucked inside a gorgeous, copper finish basket with wood handles are delicious chocolate caramel salted cookies, old fashioned vanilla caramels, honey wheat dipping pretzels and more. They're sure to be thankful for your generosity and thoughtfulness!
The Tastes of Fall Gift Basket includes: copper finish oval planter, Traditional popcorn ball, corn cob popcorn popping cop, Smokey cheese straws, 6 oz smoked salami, chocolate and caramel salted cookies, honey wheat dipping pretzels, old fashioned vanilla caramels, Chamberry truffles, grained mustard, French Onion dip mix
All prices in USD ($)
As Shown
As Shown - $72.95


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Police Arrest Nurse Volunteers At Occupy Chicago
Nurses volunteering at a first aid station for Occupy Chicago protesters were included in a mass arrest of over 100 people during a late-night crackdown at the encampment. The police officers removed all protester property from the site and tore down tents, including the first aid tent.
The action has been condemned by National Nurses United (NNU), the country's largest union of nurses, which has set up first aid stations in five other occupied cities, including Occupy Wall Street in New York.
"Even in wartime, combatants respect the work of nurses and other first responders. Yet Mayor Emanuel and Chicago seem to care as little about that tradition as they do in protecting the constitutional rights of free speech and assembly." said NNU Executive Director RoseAnn DeMoro in a statement. "These arrests are disgraceful and unconscionable, and will not deter our nurses from continuing this mission, setting up the station again, and continuing to support the protests."
On Monday morning, Occupiers showed their solidarity with the arrested nurses by joining members of NNU as they picketed the mayor's office at Chicago's City Hall.
The arrests occurred at approximately 1 am as Occupy Chicago participants attempted to rebuild their camp in Grant Park for the second time.
In many cities, police departments have been content to wave public park closing times and camping rules as long as protesters remain peaceful, respectful of non-protest pedestrians and do not disrupt the flow of vehicle traffic.
Emanuel has been perhaps the most aggressive mayor in the nation in repression of the occupy Wall Street movement with mass arrests on at least two occasions now. The Chicago Tribune Saturday reported that city officials are trying to send a message to world leaders of being "tough" on demonstrators in advance of upcoming meetings of G-8 and NATO leaders in May.
Related Reading:
Mark Ruffalo Connects Keystone XL, Fracking & #OWS [Video]
DeFazio: A Politician That Understands Occupy Wall Street
Who Are The 99%? Students Share Opposing Views
Image Credit: NNU


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CILT launch 'Stronger Together' with ITN
The Chartered Institute of Logistics and Transport (CILT) and ITN Productions have officially launched 'Stronger Together', a news and current-affairs style programme bringing to life the importance of collaboration between both individuals and businesses operating within the Logistics & Transport industry, at this year's CILT Annual Conference.
Collaboration is more vital than ever and 'Stronger Together' explores the nine sectors that CILT supports including those operating on land, sea and in the air. Membership of CILT is at the heart of the industry and supports members through knowledge, know-how and networking. This programme also includes a special animation explaining why membership is a must for anyone working in logistics and transport.
'Stronger Together' also brings a report from Core Management Logistics showcasing how value added business is changing the shape of the logistics industry. With the nations shopping habits changing, so are expectations and this report looks into how they have had to introduce a more personalised service to become an industry leader and part of their client's business.
The programme features an interview with Richard Atkinson FCILT, Head of Engagement, CILT, who talks about the need to encourage younger generations into the industry and the innovative ideas they can bring with them.
'Stronger Together' premiered on 14th June at the CILT Annual Conference and Dinner at the Chesford Grange Hotel, Kenilworth, and will form part of a wider digital campaign between CILT and ITN Productions.
Kevin Richardson FCILT, Chief Executive, CILT, said: "We are extremely pleased to once again join up with ITN Productions to produce this film highlighting the importance of collaboration within our profession. The opportunities in logistics and transport are vast, but the only way we can achieve our aims and attract bright new talent is by working together. All of our sectors need to work collectively in order to develop a profession fit for the future."
Simon Shelley, Head of Industry News, ITN Productions, said: "ITN Productions has been delighted to partner with The Chartered Institute of Logistics and Transport to bring to life how the industry needs to collaborate to achieve better results for individuals and businesses within the industry. We have enjoyed exploring what CILT is doing for their members to push the sector forward. We hope that this programme will engage and inform viewers and shape the future of such an important profession."


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Some of you might have read this and thought to yourself…
"Why would you even think I'm to blame?"
Well… It's not impossible that a woman is to blame for her Boyfriend (or Husband) cheating on her – but seeing as I have never been married I would not talk about anything that has to do with a couple bound by law and/or under God's blessing; We'll focus on couples that are just dating or courting as some may refer to it.
My close friends and family would know I am an avid lover of Nollywood and I watch a lot of their movies. A lot of them have the 'Guy cheats on girl', 'Girl cheats on guy' storyline and maybe it has influenced my thoughts – but I like to think that the messages they portray in these movies are sometimes based on real life stories or the experience of someone somewhere. So yes, who is to blame when a guy cheats on a girl – or vice versa?
Personally I feel cheating comes in all shapes, sizes and forms and I may be wrong – but this is my opinion and you have every right to agree or disagree. I have been asked a few times by friends what I would do if my boyfriend cheated on me – They know my answer – but I always start by saying cheating is in various forms. Once again I state that
"Everyone has a reason behind their action"
… but no one should say to anyone "The devil pushed me…" because he won't 'push' you if you didn't let him stay close to you!
When a person cheats I feel it's because of one of the following reasons
- They are falling out of 'love' with you
- Insecurity
- Lust
- You never meant anything to them in the first place
- Lack of 'love' and attention from you
… the list goes on but I like to think these are the main reasons. Now based on any of the above reasons, we can start to point fingers and say who is to blame. It would take a whole book to start explaining why XYZ is to blame for each of the reasons mentioned above but I really want to touch on one aspect in particular.
Why do women – especially – blame the other girl for their man's infidelity? I can understand if the girl in question was someone you know, your friend, your sister – yes sister, your cousin, colleague or just someone who knew you were dating this guy. What I DON'T understand is why some innocent women are being blamed for a man's infidelity.
I tried to think about what and how I would feel if I was seeing a guy only to find out later on that he had another girlfriend before I came into the picture – or was married. What would I do? Would it be fair on me if his girlfriend – or wife, came 'bashing' me and blaming me for his actions? I mean… I am innocent! I 'loved' this man based on what he showed me. I 'loved' him based on the man I knew. He gave me no reason to question his behaviour and because I decided to trust him I didn't bother to further investigate – he hadn't proposed or anything or should my investigations begin from the very first day he says hi? If you just said yes in your head then you are not being true to yourself because who does that? No one! Unless you are super paranoid.
Next thing – If a man manages to deceive me so well to the point that I have no clue he is seeing someone else or has another woman in his life, then that guy is SICK!. How can a human being live two lives with the single life God blessed them with? It's not normal! There is actually a name for people who have dual personalities and it is a disorder. It is called multiple personality disorder and with all due respect to anyone who may be facing this in their life, it is something that must be checked and you need some help – I say this in the most respectful and sympathetic way possible. It is not the norm for a person to live two different lives. So if you are such a guy (or girl) please get some help and get well soon!
So far the guy seems to be the person to blame yes? But if a guy doesn't have a disorder, if he is not a player and he decides to see another innocent woman… Then… Are you the cause? Are you the reason for his infidelity? Is it possible for a woman to push her man to cheat? Yes!!! It is very possible. It doesn't justify it though, but when he does access yourself too. Personally I am tired of women always saying
"He shouldn't have, even if I pushed him to it"
Girl!!! Men are humans beings too you know. If you have been a perfect… Note what I said – Perfect then I will support you 1000000000% – But no one is perfect and no one can be perfect so it all boils down to COMMUNICATION and making sure you both understand yourselves and no one is pushing anyone into anything. Even if you have been good, he may have just fallen out of love with you, maybe he isn't interested in the relationship (and guys please don't waste my sisters time – just tell her. She may cry but it is better than wasting our time. Just as guys hate the 'hard to get game' so do women hate the 'time wasting game')… Anyways – Lack of communication is probably the most common reason for cheating. You and your partner need to talk about everything. If you notice something different ASK! He may be going through something. If you cannot open your mouth to ask because you are scared, Oh girl… You need help o… But still – What's the worst that could happen? I mean, if he hits you REPORT TO THE POLICE. There is really no reason to be scared.
Sometimes, we girls also tend to neglect the obvious signs – I will not lie, I have been there too. Everyone else is seeing what is wrong, only you. What is even wrong with our brains? It just refuses to accept certain facts because of the fear of heart-break; but it is needed hunny. You need to accept it and move on – better to cry now than to cry forever.
Another problem with some ladies is that we generally tend to think we are being perfect! Being perfect girlfriends and can't seem to understand why he cheated. Ermm, are you sure or you just think you have been perfect?
There is a massive difference between how well a person is doing and how well you are actually doing. Have you ever taken a test/exam or gone for a job interview feeling so confident that you did so well, you 'smashed it' and then you get the results and you failed woefully. Yep! That's the difference. Again – COMMUNICATION– This thing is lacking a lot especially in Africa and we need our men especially to stop feeling too important and big-headed to be spoken to. Women also need to learn to calm down and find the sense in the 'non-sense' you think he is saying. Men, that a woman asks you a question, for example about your whereabouts, doesn't mean she doesn't trust you – we just like to ask – talking is our hobby… Okay?!!. I am tired of talking about cheating and infidelity but just to round-up I hope you now see that cheating can be anyone's fault.
Like I said in the beginning – Cheating comes in different forms. It's left to you to access and understand the reason and motive behind the individuals actions – whether is was the lady in the relationship or the guy in the relationship that cheated – and then make your decision from there.
Also remember that we are all human… No other comments!
Love you all. Until next week
Follow me on Instagram|| Twitter @msdemi_akin,
Subscribe to my YouTube channel High Street Mania and
Like my Facebook page High Street Mania


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The 2012 Jersey Shore HOT LIST awards are based on more than 22,000 votes from local Jersey Shore-area experts.
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GE Africa (Lagos)
GE Partners with The Tony Elumelu Foundation on Lagos Garage
GE and The Tony Elumelu Foundation (TEF) have announced a partnership that would allow up to 25 Tony Elumelu EF Entrepreneurs to participate in a dedicated 4-week advanced manufacturing training program every year at GE's Lagos Garage - a hub for advanced manufacturing-based innovation, strategy development, idea generation and collaboration. Read more. More about GE in Africa in this BRIEFING.
Nigeria: GE Partners with The Tony Elumelu Foundation on Lagos Garage
GE Africa, 7 March 2017
GE, TEF committed to skills development and growth of Nigerian entrepreneurs GE to offer an advanced manufacturing training program at Lagos Garage for up to 25 Tony Elumelu… Read more »
Mr. Tony Elumelu CON, Founder Tony Elumelu Foundation, addresses 1000 African entrepreneurs at the 2nd edition of the annual Tony Elumelu Foundation Entrepreneurship Forum, which recently held in Lagos.
The GE Garages experiences was created by GE in March 2012 to reinvigorate interest in invention, innovation, and manufacturing. Co-located with the GE Lagos offices in Victoria ... Read more »
"GE made a massive commitment to skills development and job creation in Nigeria with the launch of a permanent installation of the Lagos Garage, which is a hub for training, s Read more »


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Sane is celebrating its 25th anniversary this month and, to mark the occasion, it will be continuing the fight to beat the taboos surrounding mental illness by producing larger-than-lifesize sculptures of dogs that its founder and chief executive, Marjorie Wallace, hopes will be sponsored by companies, schools and individuals and put in prominent positions up and down the country.
Tag Archives: depression
GROUND-BREAKING free course to help carers to overcome anxiety and depression
Free carers therapy for positivity
Published on Friday 12 August 2011 09:00
A GROUND-BREAKING free course is running in Dunstable to help carers to overcome anxiety and depression.
Central Bedfordshire Council is pioneering the six-week course, which uses cognitive behavioural therapy (CBT) to help carers look at their situation and their emotional response to it.
Therapist Allison White, who runs the scheme at the local authority offices in High Street North, said: "CBT creates a positive space for carers to talk about how they are feeling and gives them some useful coping mechanisms. It's all about increasing their self worth and quality of life."
"The course provides an insight into anxiety and depression, their effects on the body, thoughts and behaviour.
The shadow of the black dog touches us all, but it is possible to master mental illness
Celebrities back 'black dog' campaign to defeat depression
Mental health charity SANE plans sculptures across the country to raise awareness of 'forgotten illness'


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Your body is smart - biologically intelligent. Biological intelligence is how your body creates and uses information. And because health is learned, you can make your body smarter. Find out how by watching this video: … [Watch the video...]
Biological Intelligence
Read more about Regeneration Health
Taking the Health Trouble Out of Air Travel
Up to one fifth of airline passengers become sick with colds or flus. Are there ways to decrease these infections? Yes. A new Finnish study … [Continue Reading...]
How Much Sleep is Too Much?
Is it true that sleeping more than eight hours a night hastens your demise? People certainly think so. Several years ago a national TV news … [Continue Reading...]
Read more about Staying Healthy
Time to Start Drinking
I'm not referring to alcohol. The summer blazes. Florida's red tide pricks at skin and distends nostrils, California forest fires clog lungs, humid urban inversions break your breath. Wet bulb temperatures of 35 … [Continue Reading...]
Eat Food, Not Vitamins
"Do you take vitamins?" "No," I answer my patients. "Why not? Aren't they good for you?" The nuanced answer to that remains no. A truthful answer might be "when it's been shown you need them." Most … [Continue Reading...]
Get Smart – Go Dirty
To prevent cancer, change early childhood education. Not the book-kind, but the biologically intelligent kind. If you do, you might change the incident of type I diabetes, asthma, peanut allergies, … [Continue Reading...]


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thing, then at least coordinated functions of the same mechanism. The stupid underground is thus both a regressive trap and a delusive utopia for those who mistake their play for a revolution that has already occurred. One could say the same for every program of social change. This bind is irreducible; there is no going beyond the delusion to reality and real political agency, any more than garden variety neurotics like yourselves can escape reality and live entirely in delusion. The empirical fact is invisible without the model, but at the same time the model eclipses it without releasing us from its demand. The map and the territory, the model and the real, the fantasy and the fact constitute each other as each other's excluded precondition. Revolutionary virtuality constitutes the very condition of the revolutionary project and guarantees its utter impossibility. The surrogate both constitutes and belies its truth, grounds it and undermines it, and the stupid underground offers a particularly stark instance of this vertiginous spiral of surrogations.
[17] What should one think of a Nobel Laureate who proposes, quite scientifically, the theory of "directed panspermia": that the nucleic proteins from which "life itself" arises were sent here from another star system? Or the notion that, since the biochemical structure of psilocybin spores resembles nothing else on earth, they too were exported, as the very seeds of consciousness, from somewhere out there? Or the proposition that language itself is a virus from outer space, or that time can be controlled by cutting up audio-tape and projecting images on top of one another? How to comprehend experiments in brain expansion through stimulation by electronic implants, or drugs; or what proposes itself as research into nanotechnology, in which tiny robots will someday patrol our bloodstreams scrubbing out cholesterol deposits and gunning down incipient cancers; or cyberprosthetics; or life extension through the ingestion of massive doses of vitamin compounds and amino acids?12 Or, to address our specific instance, what shall we make of reports of red and black rains, of frogs, fish, and highly-worked stones that fall from the sky? Charles Fort: "I have collected 294 records of showers of living things. Have I? Well, there's no accounting for the freaks of industry."13
Scenewash Kiosk |
Lobbies |
Marginalia |
Luxmachina |
It is


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It's better to miss the feeling because you can find it with someone else who will make you happy.
Seldom do we miss the person. The logical part of us can remove the person, but the way we felt while we were with them, is a memory of emotion and once they're gone we can warp that memory into perfection and never left it go. ~ Jason Krenzer
The feelings we had for them were our feelings, our heart, so it can happen again. ~ Yvonne Kertsch
Focus and get your own source of energy. Find yours and be your own master. Be independent in mind and soul. ~ Liam Bottomley
Never forget to say I love you, you never know when you may lose the chance to say it.
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The Lost Continent of Mu
Mu was an immense continent covering nearly one-half of the Pacific Ocean. When she sank during volcanic destruction, fifty million square miles of water claimed her place. This vast continent and culture was the center of civilization some 25,000 years ago. This is the story of Churchwards search for the lost continent, from the vaults of an Indian temple to the four corners of the world.
Below is a handwritten letter from Colonel James Churchward to Dr. Charles (Chas) D. Hart, dated 20 December 1932. The letter is from my personal archives having reemerged 79 years to the day that it was first written. Read Churchwards magnificent treatise: The Lost Continent of Mu.
In the letter, Churchward writes, "the Delaware Indians Chief, simply continues a symbol that was brought to this country from Mu by his forefathers …"
The 8-rayed star, known as the Sacred Symbol of Mu, was part of the Royal Escutcheon of Mu whose worldwide appearance appears cross culturally worn by the tribes of the Delaware Indians, carved in stone at Machu Picchu, etched in the Palpa sands of the Nazca Starglyph, embedded in the rock face of the Monument to Humanity at Markawasi and symbolically related to the hypercube, and tesseract.
The last vestiges of Mu (the Mother Land) are present today, reawaking in our consciousness, within our hearts.


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text: Anyone using Microsoft hosted exchange (Microsoft Exchange Online) services able to get their i1 device to sync with EAS? I'm able to integrate the EAS account on other Android OS devices (which I understand are a completely different Android OS version) such as Samsung Moment, HTC Evo, HTC Hero and even Droid X and on Windows Mobile 6 phones, however on the Motorola i1 upon entering the email address, server address, domain, username, password and select the boxes for "Use secure connection (SSL)" and "Accept all SSL certificates". I get error: " Authentication Error. Username, password and domain not match, please enter valid username, password and domain." I am positive the authentication credentials are correct since we are able to integrate on other phone models and I can complete a successful test using the Microsoft Remove Connectivity Analyzer. We have tried with and without domain as well as server address and server address IP all with the same error. Oh, and using Touchdown is not an option for us at this time. Any suggestions are appreciated.