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Swarming bacteria are challenged by the need to invade hostile environments. Swarms of the flagellated bacterium Paenibacillus vortex can collectively transport other microorganisms. Here we show that P. vortex can invade toxic environments by carrying antibiotic-degrading bacteria; this transport is mediated by a specialized, phenotypic subpopulation utilizing a process not dependent on cargo motility. Swarms of beta-lactam antibiotic (BLA)-sensitive P. vortex used beta-lactamase-producing, resistant, cargo bacteria to detoxify BLAs in their path. In the presence of BLAs, both transporter and cargo bacteria gained from this temporary cooperation; there was a positive correlation between BLA resistance and dispersal. P. vortex transported only the most beneficial antibiotic-resistant cargo (including environmental and clinical isolates) in a sustained way. P. vortex displayed a bet-hedging strategy that promoted the colonization of nontoxic niches by P. vortex alone; when detoxifying cargo bacteria were not needed, they were lost. This work has relevance for the dispersal of antibiotic-resistant microorganisms and for strategies for asymmetric cooperation with agricultural and medical implications.
IMPORTANCE Antibiotic resistance is a major health threat. We show a novel mechanism for the local spread of antibiotic resistance. This involves interactions between different bacteria: one species provides an enzyme that detoxifies the antibiotic (a sessile cargo bacterium carrying a resistance gene), while the other (Paenibacillus vortex) moves itself and transports the cargo. P. vortex used a bet-hedging strategy, colonizing new environments alone when the cargo added no benefit, but cooperating when the cargo was needed. This work is of interest in an evolutionary context and sheds light on fundamental questions, such as how environmental antibiotic resistance may lead to clinical resistance and also microbial social organization, as well as the costs, benefits, and risks of dispersal in the environment.
To thrive in heterogeneous and competitive environments, microorganisms employ both cooperative and competitive strategies. Swarming bacteria collectively migrate over surfaces driven by their flagella (1). Swarming offers a competitive advantage in invading some habitats (2, 3); however, the costs and benefits remain largely uncertain. Swarming is studied in monoculture, but more than one swarming species can associate (coswarming), with mutual benefits (4, 5). There are also instances where a motile eukaryote (e.g., Acanthamoeba spp. [6] and slugs of Dictyostelium spp. [7]) transports bacteria. Examples where bacteria provide the driving force to move sessile cargo microorganisms are rare. However, swarms of the bacterium Paenibacillus vortex (8, 9) can transport conidia of the filamentous fungus Aspergillus fumigatus (10) over long distances (tens of centimeters). The conidia gain by moving to a more favorable location, while the bacteria use germinating mycelia to cross air gaps. The idea that a bacterial swarm could be considered a moving ecosystem, incorporating sessile cargo microorganisms with costs and benefits, motivated this work. We approached this problem with respect to the contribution of antibiotic resistance by the cargo bacterium, particularly the spread of beta-lactamase (BL) genes encoding enzymes that degrade beta-lactam antibiotics (BLAs) (11).
The spread of antibiotic resistance in bacteria is of widespread concern. The BLAs include major classes of front-line therapeutics (12). Resistance to BLAs is reaching a crisis point, for example with Gram-negative pathogens having extended-spectrum beta-lactamases (ESBLs) and carbapenem resistance. Interactions between different lineages and species of bacteria are relevant to the prevalence of antibiotic resistance within a population. Marine Vibrionaceae show population-level interactions mediated by antibiotic production and resistance (13), suggesting that populations defined by a common microhabitat and an antibiotic-producing/resistance network may represent socially cohesive units. Complex relationships, including reciprocity of both antagonism and cooperation, have been found between Streptomyces strains isolated from the same soil granule (14). The idea that soil bacteria represent a reservoir of resistance genes available for exchange with clinical pathogens is supported by the discovery of identical antibiotic resistance cassettes, including ones active against BLAs, in soil bacteria and human pathogens (15). Taken together, these observations suggest that networks of microbial interactions within the environment are important in the emergence and spread of antibiotic resistance.
Bacteria on surfaces may be in physiological states that confer tolerance to antimicrobials, notably within a biofilm, but also when swarming (1, 16). P. vortex shows a limited phenotypic resistance to a diverse range of antibiotics when swarming (17). Multiple strategies exist within isogenic populations of bacteria to temporarily protect themselves from antibiotics (16, 17, 18), including ones linked to swarming. For example, swarming Bacillus subtilis undergoes spatial and population shifts between motile and nonmotile cells, with the net effect that both colony expansion and growth are less vulnerable to antibiotics (19). Salmonella enterica serovar Typhimurium has decreased permeability to some antibiotics when swarming (20). Further, phenotypic tolerance of antibiotics by P. vortex is due to a highly motile but poorly reproducing subpopulation, the explorer morphotype (17). Despite a degree of antibiotic tolerance, swarming P. vortex cannot thrive (i.e., disperse and expand their population) in the presence of high levels of BLAs.
Here, we show that swarming P. vortex can transport a different species of bacteria (cargo). When the cargo expresses a BL, transport opens up new niches for P. vortex. In the presence of a BLA on an agar plate, both transporter and cargo species gain from moving as a consortium in an asymmetric but mutually beneficial relationship: the former drives dispersal and the latter detoxifies. Additionally, P. vortex employs a bet-hedging strategy; when this bacterium invades new territory, it does so first with swarming cells without cargo bacteria and subsequently with swarming groups containing cargo. Therefore, we characterize the relationship as a mutualism of convenience, in which P. vortex uses the cargo bacteria when needed.
Transport of a BL-expressing cargo by P. vortex facilitates the dispersal and growth of both species in the presence of high levels of the BLA ampicillin.When inoculated on a 14-cm-diameter Mueller-Hinton (MH) agar plate and incubated at 37°C under aerobic conditions, P. vortex was capable of swarming continuously and reaching the edge of the plate within 12 h. When P. vortex was inoculated on a plate containing ampicillin (Amp; 40 to 200 µg ml−1), limited swarming was observed (a few millimeters) without growth. When an Escherichia coli strain (VL001; see Table S1 in the supplemental material) expressing a BL was inoculated on an MH plate containing Amp, a static colony formed. In other words, in the presence of Amp neither species could use the full resources of the agar plate: the E. coli strain could not spread and P. vortex was antibiotic sensitive. However, when inoculated together (in an equal ratio of cells) on an MH agar plate containing 200 µg ml−1 Amp, the two species could colonize the plate within 72 h (Fig. 1a).
Table S1
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Figure S2
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The strain VL001 was able to swim but not swarm. To clarify the role of cargo motility, the experiments described above were repeated using a nonflagellated, Ampr E. coli cargo strain (VL003). On MH with Amp, the nonmotile Ampr strain was able to assist P. vortex in spreading toward the edges of the plates. This indicated that motility of the cargo was not required and P. vortex was providing the driving force for the dispersal process.
Fluorescence microscopy was used to observe the moving microcolonies, after allowing the bacteria to swarm across regions of agar impregnated with fluorogenic dyes to allow distinction of the two species by selective staining (hexidium iodide for P. vortex [red fluorescence] and Syto 9 for cargo [green]) or by using green fluorescent protein (GFP)-expressing cargo. Microcolonies (hundreds of microns across) at the periphery of an expanding colony contained both cargo and transporter strains (Fig. 1b). Cargo bacteria were patterned by the transporting P. vortex (Fig. 1b and c). On MH agar plates with 200 µg ml−1 Amp, the colony was composed of concentric rings of bacteria (Fig. 1a). Colony expansion was periodic (Fig. 2), with active swarming alternating with periods of little progress. The periods of reduced expansion apparently coincided with detoxification of the antibiotic by the BL-expressing cargo, followed by swarming when the BLA remaining at the colony periphery was no longer harmful to P. vortex. This conclusion was supported by the observation that after a pause of the peripheral swarming cells, adding additional Amp (to a local concentration of >200 µg ml−1) at the edge of the colony delayed the reinitiation of swarming by 1 to 2 h. Further, addition of the BL inhibitor clavulanic acid (>0.1 µg ml−1) also delayed the initiation of swarming for 3 to 4 h. In contrast to the Ampr (ampicillin-resistant) cargo strain VL001, an Amps (sensitive) strain of E. coli (VL002; see Table S1 in the supplemental material) was unable to facilitate colonization of MH agar plates containing 200 µg ml−1 Amp when coinoculated with P. vortex. Additionally, the periodicity of swarming decreased with higher concentrations of ampicillin in the plate (see Fig. S1 in the supplemental material). Taken together, these observations suggest that the time taken to detoxify the antibiotic was the limiting factor in colony expansion.
Figure S1
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Changes in populations during the interaction of swarming P. vortex cells with Ampr or Amps cargo strains of E. coli were quantified by selective viable counts. When inoculated alone on MH agar without Amp, both E. coli strains expanded their populations by nearly 2 orders of magnitude by static growth (Fig. 3a). On the same medium, a monoculture of P. vortex achieved a major population expansion (over 4 orders of magnitude) by swarming and growth. When each of the three strains was inoculated as a monoculture on plates containing Amp, the Ampr E. coli strain expanded its population due to local growth, while the viability of P. vortex and Amps E. coli strains declined (Fig. 3b). When the Ampr cargo strain was coinoculated with P. vortex in the presence of the swarm inhibitor 0.5 mM p-nitrophenylglycerol (PNPG) (9, 21), both species gained moderately from localized growth. In this situation, P. vortex benefited from close proximity to the antibiotic-degrading E. coli. When E. coli Ampr was coinoculated with P. vortex under swarming conditions on medium containing Amp, both species expanded their population to a similar extent, around 3 orders of magnitude (Fig. 3b). The E. coli Amps strain could not support the same population expansion when coinoculated with P. vortex on medium containing Amp (Fig. 3b). Finally, when inoculated together without Amp (P. vortex and Amps or Ampr E. coli), the cargo still gained from the association: the colony expanded and E. coli was transported and grew in new regions of the agar. However, Ampr E. coli gained less from being a cargo, relative to the situation in the presence of Amp (population increase almost 2,000-fold with Amp and only 400-fold without), and P. vortex gained more from the situation (population increase 1,500-fold with Amp and 7,000-fold without).
Local analysis of cargo distribution during transport with and without a BLA.The distributions of the cargo and transporter strains were analyzed by isolating cells from different locations within the spreading colonies. Inoculated plates were sampled after 50 to 72 h, capturing >107 CFU, and the species composition was determined by selective viable counts. The expanding colony, typically 6 cm in radius, was divided into three sampling zones: the periphery of the colony (up to 1 cm from the edge), the immediate interior (1 to 2 cm from the edge), and the central region (>2 cm from the edge, including the inoculation site), as shown in Fig. S2 in the supplemental material. This analysis was particularly revealing for the situation in which the cargo and transporter strains were coinoculated on MH agar without Amp. Moving colonies at the periphery of the colony contained no detectable cargo bacteria. Both cargo and transporter bacteria were present within the colony interior (see Fig. S2). In contrast, analysis of the species distribution after coinoculation on MH agar with Amp revealed that the cargo strain was highly represented in all zone samples, including the edge of the colony. These data, combined with selective viable counts from entire plates (Fig. 3), indicated that the presence of Amp favored the transport of the E. coli Ampr strain and that this selection pressure had the greatest effect at the colony edge. The peripheral colonies lacking cargo in the absence of Amp were smaller, faster, and more highly dispersed than the interior cargo-carrying colonies (see Table S2 in the supplemental material). Microscopy showed that the peripheral colonies were derived from single cells and small groups of P. vortex cells that made an early escape from the central mass. Analysis of expanding colonies at earlier stages in growth indicated that the first wave of moving colonies could be detected after 20 min to 7 h, and these colonies (>90%) lacked cargo. A first scenario was created, where the transporter and cargo initiated swarming in a region of the plate and then the consortium was forced to encounter localized Amp after swarming 4 cm (Fig. 1d; see also Fig. S3 in the supplemental material). Tracking the moving colonies indicated that those lacking cargo (judged by microscopy imaging via GFP and confirmed by viable plating) reached the region of antibiotic first (by 1 to 2 h) and that these then failed to grow. After 3 to 6 h, the region was occupied by larger swarming colonies that contained cargo and which were able to grow.
Figure S3
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Table S2
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In a second scenario, serial culture on agar plates was used to assess the robustness of the cargo-transporter system. Inoculations with P. vortex and a cargo strain were made on one edge of a 14-cm-diameter MH agar plate, and >108 CFU were isolated from the far edge of the plate after 36 to 80 h, to compare plates with and without Amp (see Fig. S3 in the supplemental material). When Amp was absent, it took 1 to 4 transfers (10 replicates; average, 1.8 transfers) to eliminate the cargo, as judged by selective plating. When Amp was present, the system was stable, as after >20 transfers the cargo continued to be highly represented (30 to 65% of the population by viable count; n = 6). A third scenario was created in which exposure to Amp was periodic: the consortium initiated transport in the absence of Amp, then passed through a region of agar containing Amp and was recovered from the far side of this chemical barrier (Fig. 1d; see also Fig. S3). As with the situation with Amp present throughout the agar plate, the cargo was stably maintained (>20 transfers; n = 6). This suggests that intermittent exposure to a BLA is sufficient to maintain the association of the Ampr cargo and P. vortex.
Comigration of P. vortex with E. coli Ampr on plates with Amp led to a stable mixed population, a system that corrected for initial imbalances in the inoculum. When the cargo was inoculated in as little as a 1:1,000 minority (5 × 107 CFU P. vortex, 5 × 104 CFU of Ampr E. coli) or in up to a 20-fold excess (109 CFU E. coli), the result after 72 h was a final population close to parity (with up to a 3-fold excess of cargo or transporter). This, along with the long-term robustness of the system, suggests that population levels move rapidly toward equilibrium as long as the selection pressures for both colony expansion and Amp resistance are maintained. The exception was at high cargo ratios (>20-fold cargo excess; >109 CFU), a situation in which swarming usually failed to initiate due to cargo overgrowth. However, in the cases (<20%) where swarming occurred when there was a 20-fold cargo excess, the consortium formed a stable and apparently balanced population after 72 h.
Transport is mediated by a phenotypic subpopulation of P. vortex.Previous studies have shown that an isogenic P. vortex colony contains two subpopulations: explorers with elevated antibiotic resistance and enhanced motility, and more rapidly dividing but less motile builders that form the majority of the colony interior (17). These two phenotypic variants of isogenic cells were enriched separately to >99% purity (see Fig. S4 in the supplemental material) and tested for their ability to transport an E. coli cargo (compared to the mixed population, which was ~30% explorers) (17). Transport assays were run for 12 h after inoculation with E. coli (Ampr) on LB agar plates containing 100 µg ml−1 Amp. This allowed the majority of the population to be maintained as the inoculated morphotype (17). Neither species could disperse or grow without the cargo—the limited refractory nature of explorers to antibiotics was not sufficient to support swarming on this medium (Fig. 4b to d). The explorer-enriched subpopulation mediated transport and the builders did not (Fig. 4). During transport, flagella create tangled networks, apparently wrapping around the cargo (10). One of the differences between the subpopulations is a higher expression of flagella in explorers versus builders (17). Thus, a possible explanation of the explorers' ability to transport cargo could be more flagella enhancing one or both of motility and cargo capture.
Figure S4
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Cargo bacteria with higher BLA resistance support more effective dispersal on BLA-containing plates.An isogenic collection of cargo strains, differing in their levels of BLA resistance, were tested for their ability to complement P. vortex when swarming in the presence of cefotaxime (Ctx). These strains expressed a TEM-1 type BL, with the encoding gene containing mutations leading to the production of enzymes with different levels of activity (22), and consequently had a wide range of resistance to Ctx, with MICs from 0.03 to 512 µg ml−1. P. vortex was highly sensitive to this BLA (MIC, 0.06 µg ml−1) and could not grow or swarm in the presence of Ctx. As in previous experiments, E. coli alone could grow on MH agar with Ctx if the MIC was sufficient, but could not swarm. Coinoculation of different cargo strains with P. vortex on Ctx plates and incubation for 72 h indicated that the higher the MIC of the cargo, the more effectively the consortium grew and dispersed (Fig. 5a). A higher resistance level was as beneficial to the transporting P. vortex as was the BL-producing strain of E. coli, as long as the two strains had the potential to form moving consortia (Fig. 5a to c). The benefits in terms of population expansion and dispersal were greater than for the E. coli in monoculture (Fig. 5b and c). The consortia appeared more responsive to an increase in MIC of the cargo than E. coli alone, when the MIC was higher than the concentration of Ctx in the agar (Fig. 5b and c). This may be because in the static situation, E. coli only detoxifies local antibiotic, and once that is achieved it gains little additional benefit from a more effective BL. In the moving consortia, the leading edges are expanding into new territory containing fresh antibiotic. This has the potential to happen rapidly—P. vortex can transport cargo at a rate of >1 cm h−1—so we suggest that there is a greater selection pressure for rapid degradation of the target antibiotic.
P. vortex has superior transport capacity compared to other swarming bacteria.Other swarming bacteria were tested for their ability to transport Ampr cargo. Vegetative and swarmer cells of Amps strains of Proteus mirabilis failed to transport Ampr E. coli, as judged by swarm expansion and GFP visualization by fluorescence microscopy. These experiments were performed on MH Eiken agar (0.6% to 1.8%, wt/vol) plates containing 40 to 200 µg ml−1 Amp. Up to 72 h after inoculation, no expansion of the colony was observed beyond a few millimeters from the inoculation point. Ampr strains of P. mirabilis swarmed, but again, transport was not found. Similarly, a clinically derived Amps strain of Enterobacter aerogenes was able to swarm on MH agar (0.8%, wt/vol) but could not transport the same cargo. Other Paenibacillus spp. swarmed on MH agar. A collection of these were tested for their ability to employ Ampr cargo to disperse and grow on MH agar containing Amp (100 µg ml−1). Coinoculation with the cargo strain allowed localized growth of both Paenibacillus spp. and E. coli within the inoculation region, indicating that the former could benefit from a nearby BL-producing strain. Paenibacillus alvei, Paenibacillus polymyxa, and Paenibacillus glucanolyticus had minor transport capacities. Limited transport by Paenibacillus lautus was possible, as judged by the expansion of colonies on Amp-containing plates (Fig. 6) and by the distribution of the cargo strain, assessed by fluorescence microscopy. Paenibacillus dendritiformis has two phenotypic variants on agar. These morphotypes segregated stably, with rare interconversion, as the T (tip-splitting) and C (chiral) morphotypes, with different colony morphologies (23). The C morphotype transported most effectively. None of the Paenibacillus spp. tested could match the transport capacity of P. vortex. Further, the intricate patterning that P. vortex created with the cargo organism (Fig. 1b and 7) was absent with other Paenibacillus spp. The effectiveness of transport by P. vortex could be related to its swarming strategy, which involves multilayers of cells in which cargo microorganisms become stably entrapped by P. vortex, apparently within a network of flagella (10). Modeling of P. vortex "traffic systems" during cargo transport (24) supports physical connections between formed transporters in the context of a highly organized traffic network of cells (25) in which individuals maintain the same neighbors over a period of at least minutes.
Clinical isolates expressing ESBLs gain from transport.To investigate whether the P. vortex transport system can aid the spread of antibiotic-resistant pathogens, a collection of clinical isolates (see Table S3 in the supplemental material) of Enterobacteriaceae, both sensitive and resistant to cephalosporins, including Ctx (the resistant group was classified as ESBL producers), were tested as cargo organisms. This collection of resistant strains all complemented P. vortex in colonizing 14-cm MH agar plates containing 3 µg ml−1 Ctx (see Table S3). Ctxr clinical isolates of E. coli, nonmotile, capsulated Klebsiella pneumoniae (26), and Enterobacter aerogenes were all transported by P. vortex, and those that had appropriate BLs allowed transport and cocolonization of plates containing BLAs (Fig. 1c and 7; see also Table S3). Sensitive strains did not complement swarming of P. vortex on Ctx-containing agar. The motility of cargo bacteria was tested in the absence of P. vortex, and most were found to be nonswarming. One E. coli strain (see Table S3) was able to swarm under the test conditions. Even for this strain, the presence of P. vortex was still an advantage in the spreading of the consortium.
Table S3
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Environmental isolates expressing BLs gain from transport.To test the ecological relevance of a transport strategy involving BLA-resistant cargo, bacteria isolated from arable, industrial, and woodland soils were screened for isolates capable of complementing P. vortex, i.e., allowing the latter to swarm in a sustained fashion on MH agar plates containing either Ctx (3 µg ml−1) or the carbapenem BLA meropenem (Mer; 2 µg ml−1). Candidate cargo strains recovered from the edge of MH agar plates containing Ctx or Mer were purified and retested by combining them with fresh P. vortex. This resulted in the isolation of Gram-negative bacterial strains which acted as useful cargo and so enhanced the spread of P. vortex on plates containing one of these BLAs. Identification by 16S sequencing revealed that most of the Ctxr isolates were pseudomonads with Ctx MICs of >8 µg ml−1 and Stenotrophomonas maltophilia (see Table S4 in the supplemental material). The S. maltophilia isolates were resistant to imipenem but not when this BLA was supplemented with EDTA. This suggested that the primary BL that was expressed was a metallo-BL (27). Therefore, non-TEM-1-type BLs can complement P. vortex swarming on BLA. The Mer screen resulted in a different range of isolated bacteria, predominantly Flavobacterium spp. All of these groups of bacteria have been encountered within hospitals and identified as causing opportunistic infections that are particularly hard to eradicate due to their broad spectrum of antibiotic resistance (28, 29). All isolates yielded satellite colonies when plated with a BL-sensitive strain of E. coli (VL002), indicating that they could degrade BLAs. The isolation of BLA-resistant cargo organisms directly from the soil in these experiments suggests that P. vortex can employ environmental bacteria as useful cargo.
Table S4
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When Paenibacillus vortex swarms, it can carry other microorganisms for many centimeters. The cargo microorganisms may be useful in solving problems that P. vortex cannot overcome alone. In this work, we have described a widely applicable selective system for looking at cargo-transporter interactions, one that requires dispersal across an agar plate in the presence of an antibiotic for the transporter and cargo to maximize their populations. We used BLAs to prevent P. vortex from growing or swarming alone and BL-expressing cargo unable to disperse. Together, a consortium of the two strains could colonize a BLA-containing agar plate to mutual benefit as long as the selective pressure was maintained. Colonies of BL-expressing bacteria can protect nearby sensitive strains, i.e., the phenomenon of satellite colonies. The situation we have described here is more dynamic and with reciprocal benefits during expansion of a mixed population. In other ways, P. vortex gained more from the relationship. The cargo bacterium benefited by dispersal, but alone it would have grown near the inoculation site, albeit to a lesser extent than if transported by P. vortex. The latter, when inoculated onto Amp alone, suffered a population decline due to the effects of the antibiotic. P. vortex was inoculated alone and with Ampr cargo on Amp-containing plates. When inoculated alone, the viability of P. vortex declined by over 4 orders of magnitude; with cargo assistance, P. vortex gained ~108-fold CFU/ml (Fig. 3). Ampr cargo gained 100-fold CFU/ml in the presence of the transporter.
A swarming colony of P. vortex contains a highly flagellated subpopulation (explorers) that localize toward the periphery and help to expand the colony, with builders (a less motile but rapidly dividing phenotype) dominating the interior (17). A P. vortex colony can be considered a temporarily multicellular organism with differentiation and task distributions (30–33). Only explorers mediate transport; therefore, we conclude that the explorers are specialized in movement, both movement of their own cells (motility) and of others (transport), the latter being a type of bacterial task distribution not previously reported. Further, of the Paenibacdillus spp. tested, only P. vortex created intricate patterns with the cargo. P. vortex operates in a self-organizing system, but we have shown here that when it is swarming it also can act as an "other-organizing" system (Fig. 1c and 7). Ecosystem patterning is of interest at a macroscopic level (34) and also within a developmental context, i.e., how different cell types organize to form tissues (35). P. vortex appears to provide an important model for studying how cells form more complex structures, as a system with two phenotypes with different spatial distributions (explorers and builders) and the explorer subpopulation organizing other bacteria.
In the absence of antibiotics, the Ampr cargo was still transported by P. vortex, but it was absent from the colony periphery. The faster, smaller lead colonies running ahead of the rest on MH agar did not contain cargo. In this situation, the cargo was transported by a second wave of larger, slower-moving colonies lagging behind the lead elements. This contrasted with the results of the experiment when Amp was present and where the cargo was invariably found at the edge. This has an interesting consequence: swarming colonies solely composed of P. vortex get a chance to explore and exploit new territory first in the absence of selection pressure for the presence of potentially competing cargo bacteria. However, if the lead elements cannot thrive, the second rank carrying the cargo may catch up and allow the swarm to do better. In this sense, P. vortex appears to exploit the cargo bacteria. The latter appear to be treated as an equal partner only when needed. This can be considered a bet-hedging strategy: hitchhikers are only given the best ride when useful to P. vortex.
An important question is how the degree of antibiotic resistance of the cargo affects the survival of both cargo and transporter in the presence of a BLA. A higher level of BL resistance not only promoted local cargo survival but also facilitated dispersal by P. vortex (Fig. 5). High levels of antibiotic resistance made little difference to the growth of the E. coli strain on Ctx alone when the MIC was higher than the concentration of the BLA. A static colony need only survive local BLA, and once this is done it only needs to degrade BLA on the edge of the growing colony. In contrast, a cargo strain expressing a BL can be moved rapidly by P. vortex, in excess of 1 cm h−1. In that situation, the demands for rapid degradation of the BLA are likely to be higher, as the moving consortium encounters fresh antibiotic continuously. Modeling studies support the role of antibiotic gradients in promoting the evolution of resistance in bacteria (36). Transport of cargo bacteria may be one situation where such a selection pressure is generated.
Because clinical BLA resistance is a major issue, we tested isolates obtained from human infections (expressing ESBLs) as cargo. The BL-expressing strains, even an independently swarming strain, showed enhanced dispersal when combined with the more vigorously swarming P. vortex on BLA-containing plates. Additionally, a screening of soil bacteria was undertaken to isolate cargo bacteria which complemented P. vortex when coinoculated on MH agar, also containing BLAs. This resulted in the isolation of a collection of Gram-negative, BLA-degrading bacteria, including ones with BLA resistance profiles commonly found in opportunistic hospital infections (29). Further, when presented with a mixed cargo population (sensitive and resistant E. coli) in the presence of a BLA, P. vortex retained the resistant strain more so than the sensitive strain. Therefore, from a mixed population, the most useful strain (to P. vortex) was retained. This suggests that P. vortex may play a role in the (re)distribution of clinically relevant BLA-resistant bacteria in the environment. BLs are the enzymes responsible for the most prominent and one of the most clinically relevant antibiotic-degrading activities. However, other antibiotic classes, both natural and semisynthetic, can be metabolized by bacteria from the environment, and other interspecies interactions can affect antibiotic survival (37, 38). Therefore, it is possible that other antibiotic-protecting bacteria may be productively transported by P. vortex. Recently, P. vortex was shown to disperse pathogenic species of Xanthomonas spp. on tomato leaves (39), increasing infection. In this case, the less motile "cargo" stimulated P. vortex motility with unknown volatiles, suggesting a role of interspecies communication in forming swarming consortia. Different cargo microorganisms may bring other "skills" which P. vortex lacks, and P. vortex may transport such microorganisms under other types of selection pressures, driving associations in which both partners gain an advantage. Range expansion is a powerful driver in evolution (40) and here may promote interactions between the sessile and motile microbiota. Further work could productively explore the genetic basis of this phenomenon, in both cargo and transporter, and the evolutionary and ecological importance of such relationships, i.e., the extent to which swarming bacteria act as moving ecosystems. Long-term coevolution experiments of transporter and cargo combined under different selection pressures may prove particularly useful in constructing stable or metastable synthetic ecologies (41). Such constructed or artificially evolved consortia are likely to be valuable in examining cooperation, competition, and cheating at a fundamental level (42).
Strains and culture conditions.The strains used in this study are summarized in Table S1 in the supplemental material. P. vortex and other Paenibacillus species (P. dendritiformis, P. polymyxa, P. glucanolyticus, P. alvei, and P. lautus) were grown, as previously described, on MH agar (1% [wt/vol] Eiken agar) (8), Luria broth agar, or peptone agar (17), aerobically at 20 to 37°C. Cargo strains were grown on MH agar at 20 to 37°C. Proteus mirabilis was grown on MH agar (0.8 to 1.5% [wt/vol] Eiken agar) at 37°C (43).
Cargo transport assays.For cargo transport assays, 5 × 107 cells of each species (P. vortex or another candidate transporter and/or cargo organism) were inoculated into the center of an MH agar plate (1% [wt/vol] Eiken agar) supplemented with appropriate antibiotics and incubated under aerobic conditions at 20 to 37°C. Cargo transport assays with the Ctx series of E. coli strains (see Table S1 in the supplemental material) were performed in the same way with 50 µM isopropyl β-d-1-thiogalactopyranoside (IPTG) to induce the TEM-1 BL (22). Selective plating of wild-type P. vortex was on MH agar containing 1 µg ml−1 kanamycin and 0.5 mM of the swarming inhibitor PNPG with incubation at 20°C for 25 h. Selective plating of P. vortex Rifr was on MH agar with 3 µg ml−1 rifampin (Rif) and 1 mM PNPG (8, 21), and plates were incubated at 20°C for 25 h. Selective plating to determine the viable count of E. coli was on MH agar supplemented with 1 µg ml−1 vancomycin with incubation at 37°C for 18 h.
Passage to extinction experiments.To assess the robustness of long-term transporter-cargo interactions, coinoculations were made as described above, but at 1 cm from the edge of a 14-cm-diameter MH agar plate containing appropriate antibiotics (see Fig. S2 in the supplemental material). Recovery of swarming bacteria was achieved by harvesting >108 CFU from the opposite side of the plate. These samples were subjected to selective viable counts to assess the numbers of cargo/transporter organisms and reinoculation on a new plate to repeat the process. The number of passages required to eliminate the cargo was assessed. In a variation of this assay, four antibiotic-containing Neosensitabs (Roscoe Diagnostics, Taastrup, Denmark) were placed between inoculation and recovery points to create a continuous region of antibiotic-impregnated agar that had to be traversed to reach the recovery point.
Enrichment and assays with P. vortex subpopulations.Both explorer and builder subpopulations were enriched as previously described (17). Briefly, enrichment for builders was by growth in 2 ml LB overnight at 37°C. The overnight culture was diluted 1:100 into fresh LB and grown for 4 h; serial cultivation was repeated for three more passages. For explorers, cultures of P. vortex were cultured in LB containing 60 µg ml−1 kanamycin for 18 h. Verification of the two enrichment procedures is given in Fig. S3 in the supplemental material.
MIC determinations.MIC values for antibiotics were determined on MH agar (1% [wt/vol] Eiken agar) containing a 2-fold dilution series of the appropriate antibiotic, using EUCAST breakpoints and procedures to determine sensitivity and resistance (Clinical breakpoints for bacteria, version 3.1, 11 February 2013; http://www.Eucast.org/clinical_breakpoints/).
Staining and imaging of swarming cells.Staining cells with the fluorogenic dyes hexidium iodide and Syto 9 was also used to distinguish P. vortex from Gram-negative cargo bacteria. To stain moving colonies, 4-mm-diameter areas of agar 1 cm ahead of the moving colony were impregnated with 30 µM Syto 9 and 15 µM hexidium iodide (Life Technologies, Bleiswijk, Netherlands) in a 10-µl aliquot of sterile water and allowed to dry into the agar. Bacteria were stained as they swarmed into this region. Imaging was done with a BX-41 fluorescence microscope equipped with a Kappa 8 b, black and white charge-coupled-device camera and 4× and 10× Fluorotar objective lenses (8). Swarming on MH agar plates was monitored with a Dino-Lite AM7031MT USB microscope with Dinocapture software (version 1.3.6K) and by imaging plates on a heated slide warmer at 37°C through the agar. Illumination of whole plates to detect GFP expression was performed with a 470-nm LED (Thorlabs, Dachau, Germany). Calculations of swarm rates and colony diameters were made using the ImageJ software package, version 1.45s (44).
Isolation of environmental microorganisms.Arable and woodland soil samples from locations within 1 km of Utrecht (Netherlands) or within 1 km of Rotterdam harbor (Netherlands) were screened for cargo bacteria that could assist P. vortex in colonizing plates containing BLAs (3 µg ml−1 Ctx or 2 µg ml−1 Mer). Soil (5 g) was soaked in 50 ml MH broth for 16 h. After settling, samples of the supernatant were mixed with equal volumes of P. vortex (108 CFU in a 50-µl inoculum) in MH broth and incubated at room temperature for 3 days on 14-cm-diameter MH agar plates containing the appropriate BLA. Where there was migration toward the plate edge, the bacteria were isolated and streaked to single colonies on MH agar containing the same antibiotic and 1 mM PNPG to inhibit swarming. Isolates that were not P. vortex (on the basis of swarming behavior and colony morphology) were identified by 16S sequencing (45). Purified strains were tested for antibiotic resistance (MIC, ability to swarm alone, and ability to complement P. vortex on MH agar containing Ctx or Mer, as appropriate). BL activity was assessed by nitrocefin hydrolysis (Fluka, St. Louis, USA). A general bioassay for the hydrolysis of Ctx was performed, based on the growth of sensitive satellite colonies around the resistant strain. An indicator strain, a BL-sensitive strain of E. coli expressing GFP (VL001), was spread plated (ca. 106 CFU per plate) on MH agar containing 3 µg ml−1 Ctx. The candidate BL-producing strains were then streaked on the same plates. Those isolates that promoted the growth of the indicator strains within a few millimeters, after 2 days at 25°C, were scored as BLA degrading (see Table S4 in the supplemental material).
This work was funded in part by EU FP7 program EvoTAR (C.J.I.), the Tauber Family Funds, the Maguy-Glass Chair in Physics of Complex Systems at Tel Aviv University, by the NSF Center for Theoretical Biological Physics (grant no. PHY-1308264), and the Cancer Prevention and Research Institute of Texas at Rice University (E.B.J.).
The funders played no direct role in this work and were not involved in the decision to publish.
Thanks to Arjan van de Visser for strains and Rob Willems and Willem van Shaik for the supply of antibiotics.
- Received 15 January 2015
- Accepted 13 April 2015
- Published 12 May 2015
- Copyright © 2015 Finkelshtein et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.


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True Experiences of Homeless Students at BHS
"Most adults would be in a fetal position if they would have to go through half of what these kids have experienced," states Geiss, Blaine High school's social worker.
Visualize coming home tonight from school. You ask your Mom and Dad to pay the fee so you can join the sports team, but they are trying to pay the apartment rent, medical bills, and for car repairs on time, so they are tight on cash. Your Mom gets into a fight with your Dad about it, and she forgets to make you dinner. You are too stressed to eat anyway, but you can't focus on your homework, either. It's hard to fall asleep too.
The next day at school it's rough. Your teachers are disappointed that you didn't finish your homework. After having not enough sleep, you fall asleep in class, and you are too hungry to focus anyways. Your friends notice you are in a cranky mood and avoid you.
For most students, they might have a bad day or two, but they know they will come home to a warm dinner and a roof over their heads at the end of the day. "For us, there might be a temporary falling apart, but for them, there is no 'tomorrow will be better," explained Geiss, Blaine High School's social worker.
"Them" being students without permanent homes and little or no food security. So far this year, Blaine has about 50 to 60 families with low food security and 10 to 15 kids who are truly homeless.
Not many students are homeless as a result of drugs or alcohol abuse. Most end up in their situation because of a bad turn of events, like a loss of a job, an eviction (which makes even it harder to buy a new apartment) a broken-down car, trying to pay piles of medical bills, or even a house fire.
When their families usually live paycheck to paycheck, it causes their delicately balanced life to crumble. Sometimes, parents might divorce and find themselves unable to fully support all of the kids separately.
Other times, it is the student's choice to leave. If their parents are constantly fighting, they might not feel safe at home, and run away.
Whatever the cause, Ms. Geiss wanted to make it clear that it was "always outside of the control of the kid."
If a student is homeless, a friend or relative might take them in, and they'll 'couch hop' from home to home. Others sleep in their car, or in a hotel. The worst-case scenario would be on a bench or in a park; which is difficult to imagine in freezing midwinter nights in Minnesota.
"Most adults would be in a fetal position if they would have to go through half of what these kids have experienced," states Geiss.
At school, it's hard for homeless students to focus on their work when they are concerned about core needs to survive that aren't met and are out of their control. "They do their best, but their education is still impacted," says Geiss.
During a teacher's lecture, they are worrying about if they'll be able to eat dinner tonight, wondering about where they'll spend the night and if they'll get a ride there, and agonizing about their unclear future.
They are also faced with questions, like, What address should I write on this form when I don't have a permanent home? How can I give out my contact info if I don't own a cell phone? These may seem to be minor issues, but for Geiss, when a homeless student doesn't come to school, she can't get in touch with them. Some homeless students have even gone missing before.
All this built up stress causes the students to react in different ways, Geiss explains. Some students will have their head down during class, speak rudely, they might be frustrated and even walk out of class. Geiss asks that students should have "a broader view of the situation" if they see someone acting like this, "If someone is crabby or rude, you don't know what they might be going through."
Most homeless students, however, blend in the student body. Geiss says, "You can never tell who is homeless, as they work really hard to present themselves."
"There are kids that you talk to every day that you have no idea of what struggles they go through," she says.
Homeless students are often reluctant to come forward and speak about their situation because of stigma. "Their main goal is to be just like a regular student," says Ms. Pohl, Blaine's student achievement advisor. "For homeless kids, it is so hard to ask for food and support." Once, it took Ms. Pohl months to convince a student that they should accept food from Tiger Takeout, Blaine's in-house food shelf, run by Pohl and Geiss.
Around eight years ago, Tiger Takeout began in the English area, disguised as a closet. It ran on donations from the generous BHS staff, explains Pohl, "everybody wanted to be involved." As Tiger Takeout grew bigger, it was relocated in a hidden, cramped, storage room in the auditorium. New shelves were built by theater teachers who used their experience from building sets for plays. On the shelves, in addition to food items, there are personal hygiene supplies, blankets, towels, and backpacks.
Since then, other schools have followed in Blaine's example. Today, all high schools and middle schools in the district have food shelves.
Every week, volunteers come in from Tiger Takeout's faith partner, Chain of Lakes Church, whose mission is to end youth homelessness. They keep the shelf organized, and pack backpacks with food to be sneakily distributed to students, to preserve the anonymity of the homeless students.
Blaine's food shelf provides supplemental food to students and families, but unfortunately, it isn't a permanent solution to food insecurity. Although, Pohl says, "If the need is there, we will give."
Another source of relief for homeless students is Hope 4 Youth, a homeless shelter providing transitional housing for teens and families located in Anoka. Additionally, there are showers, a laundry, a food pantry, a clothing closet, and volunteers that help teens with paperwork for getting a job, receiving healthcare, finding permanent housing, and finishing high school.
Most of the students that Pohl and Geiss have worked with have successfully graduated high school. In spite of their situation, some students have even managed to get A's in their classes. "Education is a big part of their lives because it's a huge part of becoming self-sufficient adults," says Pohl.
Even students who are unaccompanied, who are not permanently connected with any adults, end up going to college. Funds from Chain of Lakes fundraisers furnish new dorm rooms for these hard-working students. For them, says Pohl, "It's the first time they'll have a permanent place of their own."
Geiss remembers a story of success coming from a former student at Blaine who was left homeless after both of her parents died. She was bounced around from relative to relative until her family wasn't able to support her. On her own, she began to work a full-time job along with school to support herself. For two years she didn't have permanent housing, and would even end up staying as far away from school as Minneapolis. Still, with "amazing strength" she graduated from Blaine, and now attends college in Duluth.
Geiss says, "It's just a situation- not who they are. It doesn't define them. They have goals and dreams too. Those kids are going to make it and grow up to be the strongest, most resilient humans."

subset: commoncrawl


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Inclusion of people with mental health disorders
Aditi Hastak, Bangalore Community Manager
Bangalore, 3 March 2016
Disability is an umbrella term for impairments, activity limitations and participation restrictions either due to physical, mental, intellectual or sensory deficiencies, of which mental illness is the most challenging. Association of People with Disabilities (APD) engages with slums in Bengaluru and other small towns to identify, build capacity and attempt social integration of people with mental health disability.
In the state of Karnataka, it is estimated that over five million people are suffering from mental illness. "Considering that a considerable part of the population is affected by mental illness in varying degrees, our main objective should be to end the stigma attached to the condition. The next step is to advocate for treatment, recovery, and rehabilitation of the affected individuals," mentions the APD website.
The APD India works across several verticals of livelihoods, education, healthcare and advocacy to empower the people with disabilities (PWD). In the vertical of health care, mental health is an integral component. This type of disability is laden with stigma, neglect, isolation, abuse and, in some cases, traditional forms of treatment. On the other hand, the provision of healthcare services is minimal, with less than 700 healthcare professionals, doctors and social workers in Karnataka dealing with such type of disabilities.
Keeping in tandem with this gap, APD commenced a community mental health program in 2004. The program has been operational for more than 10 years now. The program started with 19 slums in Bangalore and then expanded to other districts within Karnataka like Davangere, Kolar, Chikballapur and Bijapur. Recent stats record that 1,137 people with mental disorders were identified and involved in the health program in 2013-14 and 947 in 2014-15.
The program operates at multiple levels, starting from identification. A primary survey of the location for at least four to five months to understand common problems and identify people with mental illness is conducted. The identified people are then directed to a suitable healthcare facility. Though it sounds straightforward, in reality the process is an arduous task of breaking barriers of traditional healers, stigmas and sometimes lack of willingness to receive treatment.
While working with individuals to improve the situation is on-going, engagement with communities and government agencies for capacity building is also happening. Social awareness through various means of plays, exhibitions and monthly meetings to interact with parents, caregivers and residential camps are just a few of the activities that APD conducts. The final phase is rehabilitation and social integration through regular counselling and appropriate livelihood training after their condition stabilizes.
The concern of livelihood opportunity is the last issue one thinks of, and the most difficult part is to identify people with such illness, make them and their families to accept this not as a "bad" thing but a disorder which can be corrected like any other. Another major factor that needs attention is the availability of experienced and specialized professionals in taking care of such cases. An article cites that out of the five years of Bachelor of Medicine and Bachelor of Surgery (MBBS) education in India, only two weeks of clinical postings are dedicated to psychiatry. Both have to improve to adequately address mental illness issues.
Photo credit: APD India
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- "Leave this place!"
- ―Unidentified Noghri guard.
Yalara was a primitive planet located within the Yalara system of the Wild Space.
Yalara was originally inhabited by a primitive species of Yalarans that were not ready to become part of galactic society. To protect them, Jedi Master Broden Kel Verdox installed a cloaking device that masked the whole planet from the galaxy.[2]
Noghri were sent there, who wiped out the Yalarans defending the device.[2]
Battle of YalaraEdit
In 14 ABY, the Imperial Remnant, under the command of the Disciples of Ragnos, attempted to lay hands on the cloaking device. However, Jedi Scholar Tionne Solusar soon rediscovered the planet with her holocron, and, in order to thwart the Imperial plans, Jedi Knight Jaden Korr was sent to destroy the mechanism. After a short skirmish with Imperial Remnants, Jaden Korr managed ascend to the top of the tower, holding bombs to destroy the device. After a fight with many New Reborns and Noghri on the top, Jaden Korr manged to deploy four bombs around the device and successfully destroyed the cloaking device.[2]
Aftermath of the battleEdit
After returning to the Jedi Academy, Skywalker suspected that Darth Vader, who used many Noghri as slaves and bodyguards, had ordered the Noghri to exterminate the Yalarans and guard the cloaking device. The remaining Noghri on Yalara were returned to Honoghr.
- Star Wars: Jedi Knight: Jedi Academy (First appearance)


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July 26, 2016
The limits of May's freedom of manoeuvre
A month after the Brexit vote, the debate has calmed a little as political Britain is descending into its annual holidays - except for the Labour Party, which is holding a leadership election. Our interest remains focused on the dynamics of the Brexit process, and to this effect we have picked on three comments today, each focusing on different aspects.
There is now a suspicion among those who were hard Brexiteers during the campaign that Theresa May could go soft on Brexit. They don't feel that she would undo it, but rather that she'd opt for an EEA agreement, which would not allow Britain to control immigration. Melanie Philips notes in her Times column that May has already shown weakness by her readiness to compromise on the red lines of Brexit. She must stop now.
"If she fails to respect the passionate desire for self-government expressed by more than 17 million citizens, she will not only damage British interests and harm her own reputation. She may also end up destroying the Conservative party she now so unexpectedly leads."
We agree with Philips on this specific point. The real red line for the new prime minister is keeping the party together. It is hard to see that this a permanent EEA agreement could do this - though we think that a time-limited agreement, of say two to five years, leading to an FTA with the EU - would constitute a reasonable compromise that would respect the Brexit vote, and cause minimal economic damage.
William Hague is mapping out a new post-Brexit industrial strategy. He notes that the industrial revolution that is currently under way will have a big impact on lower and middle class jobs, and may drive more and more people into insurrectional opposition to the establishment. Brexit allows the UK to focus much more on investments, including public sector investments, that would give the country a niche in those markets of the future. The only way to keep the army of discontented voters happy is through strong firms, better jobs, and adaptive learning.
And finally, we noted a rather emotional comment by Marie-Noëlle Loewe, who lives in the UK, and who says that Brexit is personal:
"I, as many other mobile European citizens, have felt a bit uncomfortable. The public mood has shifted from being welcome here to being tolerated. It is hard not to take the increasingly negative language used with regards to immigrants personal. Schrodinger's immigrant, i.e. the one stealing British jobs whilst equally claiming unemployment benefits, does not exist. Ultimately, the rhetoric is aimed at those who are different, those who were not born here, those who are changing the make-up of this country through their mere presence – people like me."


text:
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The old customs hut at Pettigo. Picture: Paul Bell
While taking some time out a few weeks ago in Clones prior to the Ulster Final, I showed a flabbergasted friend how a journey from Cavan went in and out of the Republic of Ireland and Northern Ireland at three different junctures.
In his latest publication, 'Hard Border: Walking Through a Century of Irish Partition', Clones native, Darach MacDonald walks the 50-mile route along the Ulster Canal, and his experiences will resonate with any of you who live along the frontiers of Fermanagh, Tyrone or Derry.
As he reminds us: "...from my earliest age, I realised we lived in a frontier town, and these steel impediments marked the border even more forcefully than the customs posts on the Newtownbutler Road where motorists had to have their passbooks checked. Yet there were some marginal benefits of living right on the cusp of another jurisdiction, such as the border shops that offered Opal Fruits, Mars bars, Spangles and other confections unavailable on our side.
"We had television for a full decade before RTÉ began broadcasting down south. Initially, it was just BBC, but when UTV went on air from Belfast's Havelock House on Halloween 1959, we found a use at last for the rotary channel switch on the Murphy TV set.
"By then, we were thoroughly immersed in the culture, current affairs and social mores of Britain and its outpost in the north of Ireland.
"When RTÉ came along, it seemed amateurish, parochial and dated by contrast with Cathy McGowan's Ready, Steady, Go on UTV, BBC's Juke Box Jury and, of course, Top of the Pops. We were there for the gritty start of Granada TV's Coronation Street and TV hosts such as David Frost inured us against Gay Byrne's Late Late Show from Dublin."
Culture and attitudes
When he writes that "our border was always hard, because it intruded politically in communities that are almost indistinguishable, differentiating people with the same beliefs and allegiances, history and traditions, culture and attitudes" and that for "... a century, we were branded as different, corralled into political entities not of our choosing and condemned to peripheral neglect", he could indeed be writing our story.
Darach's uncle is the famous playwright Eugene McCabe and he recalls how cross-border routes were 'spiked', including at Kilrooskey, a Fermanagh townland that loops into Monaghan just north of the town, cutting off his uncle's farm. I have a memory of him building a small link road through a corner of a field to provide a circuitous escape route to town. He called it the Khyber Pass, in memory of another north-west frontier road. It is still there today, skirting the house where my maternal grandmother settled for her final years between two daughters and their growing families.
The house is divided by the border, and that proud County Clare woman relished my dad's cheeky observation that she slept 'with her head in the Free State and her backside in the North'."
As I drove through Pettigo on my return from Clones, and watched the football celebrations in the town, I thought how apt the head and backside analogy was in our own wee corner.
My own father's first station was Carrigans in 1947, so I grew up with many hilarious tales of smuggling.
Darach's experience mirrored his: "Smuggling became a popular wartime pursuit as ordinary folk played off shortages and rationing on one side against plenty and surplus on the other.
"So, for instance, tea poured south and sugar and butter went north. For Dan Kerr, it was a frantic and fun-filled period. As well as working as a young railway porter, he stood on the front line lest Crown forces attempt to invade the poorly armed Free State to prevent Germany from doing so first as a step towards an invasion of Britain."
Dan recalls: "We all had to do our training. You know, you were called out at night for mock battles and such like. We were fully qualified as soldiers, but we were all just on rations (rather than wages).
"As far as I remember, there was a half-ounce of tea per person and you got brown bread. And other things, like cigarettes, were rationed. You never saw bananas, or fruit, or anything at all like that."
"So, it was like an Ulster (Gaelic Football) Final coming out of that station, trampling each other to get up the town. And then they'd all come back and you could see them bulging with all the stuff they'd bought hidden inside their coats.
"And when they'd go back on the train to Monaghan, where the customs boarded, all the stuff would be taken off them.
"There used to be more yarns told about what was going on. Like this lady who sat down and the customs man says, 'You're very big, what have you got in there?' and she says, 'Come back in three months and I'll tell you'."
Dan also recalled being put on full alert when British soldiers from Derry strayed across the Donegal border, sparking fears of an imminent incursion!
'Hard Border' makes compulsive reading and is published by New Island Books.
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Texas Execution – Routine or "Awesome"?By
Austin, Apr. 27th—Texas conducted its fourth execution of the year this week, an event now so routine that the state's major newspapers didn't even send reporters. It was left to the Associated Press to tell the story they missed. Here in the state capital, the report was buried in the second section of the local paper.
29-year-old Richard Cobb was put to death for a 2002 murder. The United States Supreme Court had turned down his last appeal two hours earlier. According to AP reporter Michael Graczyk, when Cobb was offered the opportunity for a last statement he said, "Life is too short to harbor feelings of hatred and anger. That's it, warden."
But, as Graczyk noted, that wasn't it. After the lethal chemicals were administered, Cobb turned his head, looked up at the warden and said loudly, "Wow! That is great. That is awesome. Thank you, warden!" He then slumped and died. (I've been following death penalty cases for years, and Cobb's positive reaction is a definite first.)
The drug used in Texas is pentobarbital, a barbiturate that has been withdrawn by its Danish maker, which will not sell it for use in executions. That decision has several capital punishment states scrambling for a new supply, but the Texas Department of Criminal Justice apparently stocked up before the ban. According to Austin's American Statesman, the agency may have spent as much as $50,000 on execution chemicals and supplies.
Cobb's execution was the 496th in Texas since 1976. It is the nation's undisputed leader in capital punishment; runner-up Virginia put to death only 110 inmates in the same period. The state's pharmacy is stocked and ready to continue the pace.
Around here, it is simply routine.


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It is now certain that Mahesh Kathi's political ambitions are buried for ever. Not many know that Kathi Mahesh was looking at an active political career. He wanted to contest from some constituency in Chittoor and was approaching various political parties.
At one stage, when he was taking on Pawan Kalyan and his friend Poonam Kaur, many thought he would contest on the TDP ticket. He was seen as a counter to Pawan Kalyan. In the meanwhile, he also made common cause with Gujarat's Dalit MLA Jignesh Mevani. He met him when Mevani came to Hyderabad recently. He was also seen moving closely with Madiga groups, including IPS officer Praveen Kumar. Later, it was rumoured that he had approached the YSRCP in Chittoor district.
But, now that he is embroiled in an unwanted crisis involving Lord Ram, no party wants to touch him. He has unnecessarily made comments on Lord Ram and any party that gives him ticket will have to face the Hindu anger. So, Kathi Mahesh lost both his TV career, his career as a film critic and also his political career.


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The ground floor houses the main services: the dining room, family room and two bedrooms with playground for the children. The upper floor houses the master bedroom and its services, also a new study designed eminently to benefit and provide an area of decent work for the client.
Location: Mexico City
Area: 450 sqm
Design: 2013-2014


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[News] Is GeoHot ready to release Jailbreak for iOS 4 ?
Thursday, July 22, 2010
Posted by skygoat76 in news
A very curious post on Twitter is leading some to believe that a jailbreak release is imminent from Geohot. If so, the move would come as quite a surprise given the great fanfare that recently surrounded Geohot's alleged and self-proclaimed "retirement" from the jailbreak community - a move that saw Geohot cancel his Twitter account, set his blog to private, and assign good friend Mike Cohenn as his proxy, noting that all future messages would come through Mike.
Today, one such message from Mike was received and it has the jailbreak community on a tentative high alert. The message: What's a good time to make it ra1n? Naturally, one could be reading way too much into this particular tweet, but Cohenn's relationship with Geohot is solid and it stands to reason that the jailbreak master may have simply thrown everyone off his scent to toil in private for the last few days of the jailbreak release...


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Angie Harrod showed professionalism, kindness, and patience when confronted with a very challenging, severely and persistently mentally ill patient that showed up in her caseload in August.
Every week the patient resisted Angie's visits, he was often contentious, oppositional and unpleasant. The house was filthy and foul smelling even to this social worker's standards. The patient was unkempt, often unclothed and unwilling to accept PCS service. Always a professional, Angie persevered with patience, kindness, and respect. Week after week Angie gained ground and eventually won a tiny bit of trust.
When it came to a head as Angie predicted, she stepped up showing leadership. She called in her interdisciplinary team and provided superior care offering reassurance, kindness, and respect. The patient opposed going to the Dayton Hospice House at first, but relented and allowed the ambulance staff to take him. Because of Angie's commitment to her patient and to the mission of Ohio's Hospice of Dayton, the patient was finally made comfortable physically, mentally and emotionally. The patient's daughter, who had been estranged for 20 years, took the opportunity to drive to Dayton to say goodbye to her father. The patient died peacefully and with dignity the following day.


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Pilate Condemns Jesus
Pilate assembled the chief priests and other Jewish authorities.
Pilate: You presented this man to me as a rabble-rouser, but I examined Him in your presence and found Him not guilty of the charges you have leveled against Him. Herod also examined Him and released Him to my custody. So He hasn't done anything deserving the death penalty. I'll see to it that He is properly whipped and then let Him go.
[It was the custom for Pilate to set one prisoner free during the holiday festivities.]
Crowd (all shouting at once): Away with this man! Free Barabbas instead!
Crucifixion is a favorite Roman punishment for insurrectionists, slaves, and prisoners of war. Anyone daring to defy the power and authority of Caesar is executed in this public and humiliating way. Jesus indeed is a revolutionary. He doesn't come to proclaim a new religion, but a new kingdom—a new way of life. He is indeed a threat to Caesar's way of doing things, a way that co-opts the religious leaders.
Jesus' revolution is a peaceful revolution. He doesn't advocate the use of violence—in fact, when one of His disciples uses the sword to try to protect Jesus from arrest, Jesus heals the "enemy" and rebukes His disciple. So Jesus doesn't support the regime of Caesar or follow the usual violent path of revolution: He leads a revolutionary revolution—in a path of love, healing, justice, and reconciliation.
Jesus appropriates and transforms the symbol of their power into a symbol of His greater power. He makes the cross not the icon of violent domination, but the reverse. By hanging on the cross and speaking of forgiveness, Jesus shows that there is a greater power at work in the world than the power of domination: it's the power of God's saving and reconciling love.
Barabbas had been imprisoned after being convicted of an insurrection he had led in Jerusalem. He had also committed murder. Pilate argued with them, wishing he could release Jesus, but they wouldn't be silenced.
Crowd (shouting): Crucify Him! Crucify Him!
Pilate (countering a third time): Why? What has He done that is so evil? I have found in Him no offense worthy of capital punishment. As I said, I will punish Him and then release Him.
But they would not relent. They shouted louder and louder that He should be crucified, and eventually Pilate capitulated. So he pronounced the punishment they demanded.
He released the rebel and murderer Barabbas—the insurrectionist they had pleaded for in His place—and he handed Jesus over to them to do with as they desired (vv. 13-25, The Voice).


text:
The applicant [Mr Maurice Papon] is a French national, born in 1910 and currently in custody in the Santé Prison in Paris. He was represented before the Court by Mr Francis Vuillemin and Mr Jean-Marc Varaut, both of the Paris Bar. The respondent Government were represented by their Agent, Mr Ronny Abraham, Director of Legal Affairs at the Ministry of Foreign Affairs.
A. The circumstances of the case
The facts of the case, as submitted by the parties, may be summarised as follows.
On 2 April 1998 the Gironde Assize Court found the applicant guilty of aiding and abetting crimes against humanity and sentenced him to ten years' imprisonment. On 21 October 1999 the Court of Cassation ruled that he had forfeited his right to appeal on points of law because he had failed to surrender to custody. Those proceedings gave rise to application no. 54210/00.
From 22 October 1999 to 13 November 1999 the applicant was held in Fresnes Prison, initially in the prison hospital and then under normal conditions in a cell in the prison's solitary-confinement wing. On 13 November 1999, on an application by his lawyer, who considered the conditions in which the applicant was being held unacceptable (particularly the very low temperature in his cell), the prison authorities decided to transfer him to the Santé Prison in Paris on the ground that he could be provided there with conditions of detention that were more suitable. Since then the applicant has been serving his sentence at the Santé Prison.
1. Applicant's state of health and medical supervision
The applicant is currently 90 years old and had a triple heart bypass operation in 1996. During the investigation of his first application for a pardon on medical grounds (see below), it was noted in a preliminary expert report of 4 January 2000 prepared for the Principal Public Prosecutor at the Paris Court of Appeal, that the applicant's heart condition had deteriorated and the recommendation was made that he be immediately transferred to a specialist cardiology unit where it could be decided whether he was in urgent need of a pacemaker. He was fitted with a pacemaker on 12 January 2000.
On 2 April 2001 Dr Sicard, the head of the Internal Medicine Department at Cochin Hospital prepared a medical certificate at the applicant's request that read as follows:
"I, the undersigned, certify that I admitted Mr Maurice Papon to hospital in the Internal Medicine Department of which I am the head from 20 to 28 March 2001.
I will not go back over the patient's previous heart problems, for which he was hospitalised and provided with other medical certificates in 1999 and 2000.
The patient is currently suffering from increasingly incapacitating angina. For three months now he has found it difficult to climb the stairs (two floors) leading up to the visiting room. This difficulty in getting up stairs, which amounts almost to disablement, has prompted him to stop taking walks, especially as he now has chest pains from time to time even when resting.
Clinical examination
The patient's general condition is good; he is perfectly aware and lucid. His heart sounds are regular and his blood pressure reading is 140/80. He has some oedema of the lower limbs, probably linked to the reduced intake of diuretics, which he does not tolerate well urologically or in terms of hydration. He gets extremely tired when he is dehydrated. His pacemaker is working well and is due to be checked in a few days' time.
Laboratory test results
None of the tests reveals any renal, haematological, or electrolyte anomalies or any biochemical evidence of inflammation. The patient's anticoagulant treatment is well balanced.
Scanning results
X-rays of the lungs are normal. A cardiac ultrasound scan shows that the left ventricle is functioning normally without any pulmonary hypertension. A cardiac scintigram before and after injection of thallium after 24 hours shows that the two low-uptake areas in the posterior and anterior regions are myocardially viable, meaning that the strain on the myocardium remains partly reversible. Neither an exercise test (even mild) nor a coronarography were carried out because of the attendant risks.
General conclusions
It is likely that the coronary arteries, which have undergone a triple bypass, have become less permeable; this would explain the recurrence of angina during physical exertion.
Although there is no congestive heart failure, the patient's progressive coronary disease means that there is still a worrying risk of his suffering a fatal heart attack and the possibility of sudden death in prison cannot be ruled out, even though it is entirely possible that his heart function, which is relatively well preserved in itself, may be compatible with living a good deal longer.
It appears obvious to us, however, that keeping this 90-year-old man with a vascular and coronary disease in prison is not likely to lessen the danger of his sudden death."
According to the information provided by the Government, the applicant's health is monitored by doctors and nurses from the prison's Outpatient Consultation and Treatment Unit (UCSA), which was set up inside the prison under an Act of 18 January 1994 (it provides a doctor's surgery, twice-daily nurse clinics and weekly laboratory tests and electrocardiographic check-ups), and by specialists consulted in hospitals outside the prison (cardiological consultations in Cochin Hospital, a period spent in Jean-Rostand Hospital for a cardiovascular check-up and fitting of a pacemaker, and regular checks on the pacemaker in the same hospital).
2. Applicant's conditions of detention
The applicant is being held in an individual cell in the prison's "VIP" wing. His cell is 12 sq. m in area and is furnished with a bed, two tables, two cupboards, two chairs, a television and, at the medical service's request, a medical chair and an automatic air humidifier. There is an alarm bell at the head of the applicant's bed linked to the control centre, which is permanently manned by a warder. In one corner there is a lavatory and a washbasin; the shower is not far away and has been equipped with a seat as it does not have any grab bars. The applicant himself is responsible for cleaning and tidying his cell, with the assistance of another inmate once a week if need be.
The applicant finds it difficult to reach the exercise yard because of the steps leading to it. He has been given permission to walk along the prison landings twice a day. He has regular visits from his family and his lawyers.
3. Applications for a pardon on medical grounds
On 23 December 1999 one of the applicant's lawyers applied to the French President for a pardon on medical grounds. The application was refused on 7 March 2000 after an expert medical report had been obtained.
On 20 June 2000 the lawyer submitted a further application for a pardon on medical and humanitarian grounds, which was refused on 24 October 2000 after a medical examination by two experts on 21 July 2000. Neither of these expert reports was communicated to the applicant.
4. Letters from the applicant to the prison governor
On 19 March, 21 April and 1 July 2000 the applicant sent letters to the governor of the Santé Prison, in which he complained that the light was turned on several times a night at around 4 a.m. or 5 a.m. (letters of 19 March and 21 April) and that a warder had refused to allow him to take a walk on the landings (letter of 1 July).
B. Relevant law
1. French law
French law does not set an upper age-limit for pre-trial detention or the serving of prison sentences.
However, an offender's state of health (in which age may be an aggravating factor) may be taken into account when an application for a pardon from the French President is being considered (Articles 17 and 19 of the Constitution) or when it is being decided whether to grant parole under Article 729 of the Code of Criminal Procedure, which provides:
"Parole is designed to encourage the rehabilitation of convicted prisoners and prevent them from reoffending. Convicted persons serving one or more prison sentences may be granted parole if they have made serious efforts to readjust to society, particularly if they can show that they have engaged in occupational activities or regularly attended an education or vocational training course or have taken part in a work experience scheme or had a temporary contract of employment with a view to their social integration, or that their presence is essential to the life of their family, or that they have to undergo treatment or that they have made efforts to compensate their victims.
Subject to Article 132-23 of the Criminal Code, parole may be granted if the length of the sentence already served by the prisoner is at least equal to that remaining to be served. However, recidivists covered by Articles 132-8, 132-9 and 132-10 of the Criminal Code may only be granted parole if the length of the sentence already served is at least double that remaining to be served. In the cases provided for in this paragraph, the qualifying period may not exceed fifteen years..."
2. Comparative law
In criminal law, advanced age is not a bar to prosecution, pre-trial detention or a prison sentence in the Council of Europe's member States.
Age may, however, be taken into account, together with other factors such as state of physical or mental health, when sentences are being determined. In Andorra, in particular, the courts may, in respect of offences punishable with a prison sentence of less than three years, substitute another penalty for the custodial penalty in view of the character and the general circumstances of the accused. In three countries a sentence of life imprisonment cannot be imposed on a person above a certain age: 60 in Romania and Russia and 65 in Ukraine; in Greece life imprisonment is reduced to 16 years for people over 70.
As far as the execution of sentences is concerned, age is rarely taken into account per se but in conjunction with state of health. In Luxembourg, however, prison sentences for minor offences are not executed if the convict is over 70 (in the case of serious offences, sentences are served in a semi-open prison). Two countries (Italy and San Marino) make provision for courts to substitute house arrest for a prison sentence where the convicted person is partially disabled and over 60 (in Italy) or 65 (in San Marino). German law also provides for the enforcement of a sentence to be deferred or suspended in the event of illness.
In a number of countries age is taken into account for parole – which is generally granted sooner once the prisoner has reached a given age, namely 70 in Spain and Greece, and 60 for men and 55 for women in Romania – and for pardons.
There are also several countries in which prisoners are not required to work beyond a certain age.
Relying on Article 3 of the Convention, the applicant submitted in his initial application that keeping a man of over 90 years of age in prison was in itself contrary to that Article and that the conditions of detention in the Santé Prison were not compatible with extreme old age.
In his observations in reply he stated that it was the combination of his advanced age and his state of health which made his imprisonment incompatible with Article 3, and he drew attention to several aspects of the conditions of his detention and mentioned various incidents which he considered to be degrading and humiliating.
The applicant alleged a breach of Article 3 of the Convention, which provides:
"No one shall be subjected to torture or to inhuman or degrading treatment or punishment."
(a) The applicant complained about the conditions of his detention in solitary confinement in Fresnes Prison from 5 to 13 November.
Under Article 35 § 1 of the Convention, the Court may only deal with a case after all domestic remedies have been exhausted and within a period of six months from the date on which the final decision was taken. Where an applicant complains of a continuing situation, the period of six months set by Article 35 § 1 begins when the situation ends (see, among other authorities, Agrotexim Hellas and Others v. Greece, application no. 14807/89, Commission decision of 12 February 1992, Decisions and Reports 72, p. 148 at p. 167).
As the application was lodged on 10 January 2001, this part of it is out of time and must be rejected in accordance with Article 35 § 4 of the Convention.
(b) The applicant mentioned a number of incidents in his observations in reply. He had been struck by a warder and some of his mail, including letters from his Swiss lawyer, had been opened by the prison authorities.
The Court notes that, supposing those facts to be established, the applicant made no mention of them in his letters to the prison governor and in any event did not bring any proceedings in the appropriate courts. That being so, he failed to exhaust domestic remedies in this respect and this part of his application is inadmissible under Article 35 § 4 of the Convention.
(c) The applicant submitted that the combination of his extreme old age and his state of health meant that his detention was in breach of Article 3 of the Convention.
The Government were of the opinion that the conditions of the applicant's detention satisfied all the criteria established by the case-law relating to Article 3 and referred, in particular, to the Raninen v. Finland judgment (16 December 1997, Reports of Judgments and Decisions 1997-VIII, pp. 2804 et seq.) and the Kudła v. Poland judgment ([GC], no. 30210/96, ECHR 2000).
According to the Government, the applicant was under constant medical supervision as part of a health-care arrangement capable of dealing with any emergency. In any case, it seemed that he did not dispute the standard of the medical care provided and was not arguing that it was impossible for him to serve his sentence for health reasons but was seeking to call into question the very principle of his being in prison. He was being held in particularly favourable conditions which made full allowance for his situation, and his advanced age was not in itself a sufficient ground for finding that it was impossible for him to be kept in prison. The Government also pointed out that the complaints raised by the applicant in his letters to the prison governor had been successively dealt with and some of the problems had been solved (among other things, the chain on his fanlight had been lengthened and a seat had been installed in the shower).
Lastly, while observing that it had never been the purpose of imprisonment to improve a person's state of health, the Government argued that Dr Sicard's medical report noted the applicant's good general condition and did not conclude that his state of health was incompatible with his continued detention. In the final analysis, the Government submitted that his imprisonment did not exceed the "unavoidable level of suffering inherent in detention" and that his continued detention in the Santé Prison did not amount to inhuman and degrading treatment within the meaning of Article 3 of the Convention.
The applicant submitted that any form of imprisonment of a man of over 90 constituted in itself and in its essence a breach of Article 3 of the Convention. He cited recent parliamentary reports on French prisons which addressed the subject of old age in prison and raised the question whether the sick or the elderly should be kept in prison. He also drew attention to the fact that the former French Minister of Justice, Robert Badinter, and the main French human-rights associations had come down in favour of his release.
The applicant also pointed out that he had submitted two applications to the French President for a pardon on humanitarian grounds and that, as part of the consideration of his case, he had been examined by experts on two occasions. However, the reports had not been communicated to him even though, during the first expert appraisal, one of the conclusions of a preliminary report of 4 January 2000 had been that there was a "danger compromising the patient's chances of survival if he continue[d] to be held in detention in his cell". In the applicant's opinion, the mere fact of holding a person of over 90 in the conditions that obtained in French prisons amounted in reality, albeit not in appearance to applying the death penalty by precipitating the prisoner's death.
Lastly, in his observations in reply, the applicant complained about various aspects of his conditions of detention (the air humidifier produced stifling clouds of steam; the chain on the fanlight was too short, so that he had to climb on a chair to open it; there were no grab bars in the shower; his light was turned on at night; and he could not reach the exercise yard because the steps leading to it were too steep) and gave details of his various health problems (weak heart, lungs, and legs, and the risk of a hernia) which were worsened by the constant stress of his imprisonment. He also complained about a number of humiliating incidents and practices (body searches; police escort when he went to hospital; assault by a warder who he said had struck him; opening of mail from various correspondents, including a clergyman and his own Swiss lawyer). He also objected to the standard of his medical treatment.
In conclusion, the applicant considered that the combination of his extreme old age and his ill health made his imprisonment inhuman and degrading.
The Court observes, first of all, that the applicant is currently serving a ten-year prison sentence imposed on him by the Gironde Assize Court on 2 April 1998, when he was 88 years old. The Court's task is not to assess whether that sentence was justified but to ascertain whether its enforcement is in breach of Article 3 of the Convention.
If it is to fall within the scope of Article 3, ill-treatment must attain a minimum level of severity. The assessment of this minimum is, in the nature of things, relative: it depends on all the circumstances of the case, such as the nature and context of the treatment, the manner and method of its execution, its duration and its physical or mental effects and, in some instances, the sex, age and state of health of the victim (cf. the Assenov and Others v. Bulgaria judgment of 28 October 1998, Reports 1998-VIII, p. 3288; § 94, and see the Kudła judgment cited above, § 91).
Where persons deprived of liberty are concerned, Article 3 compels the State to ensure that a person is detained in conditions which are compatible with respect for his human dignity, that the manner and method of the execution of the measure do not subject him to distress or hardship of an intensity exceeding the unavoidable level of suffering inherent in detention and that, given the practical demands of imprisonment, his health and well-being are adequately secured by, among other things, providing him with the requisite medical assistance (see Kudła judgment cited above, § 94).
In the instant case the applicant relies essentially on his extreme old age, combined with his state of health.
The Court notes that advanced age is not a bar to pre-trial detention or a prison sentence in any of the Council of Europe's member States. However, age in conjunction with other factors, such as state of health, may be taken into account either when sentence is passed or while the sentence is being served (for instance when a sentence is suspended or imprisonment is replaced by house arrest).
While none of the provisions of the Convention expressly prohibits imprisonment beyond a certain age, the Court has already had occasion to note that, under certain circumstances, the detention of an elderly person over a lengthy period might raise an issue under Article 3. Nonetheless, regard is to be had to the particular circumstances of each specific case (see Priebke v. Italy (dec.), no. 48799/99, 5 April 2001, unreported, Sawoniuk v. the United Kingdom (dec.), no. 63716/00, 29 May 2001, unreported, and also, mutatis mutandis, V. v. the United Kingdom [GC], no. 24888/94, §§ 97-101, ECHR 1999-IX).
The Court has examined all the documentary evidence submitted by the parties. From these it emerges that, although the applicant is over 90 and has health problems which restrict his freedom of movement (in particular, heart problems as he has had a triple heart bypass operation and has had a pacemaker fitted), Dr Sicard describes his overall condition as "good" and finds that "he is perfectly aware and lucid" and shows no signs of dependence. Although the applicant objects to certain aspects of the medical treatment he is undergoing, the Court notes that he is under regular medical supervision and receives treatment both from the medical and paramedical staff in the prison and through hospital consultations and periods spent in hospital.
The Court has also examined the conditions in which the applicant is being held. While it is certain that he is not enjoying the same quality of life as he would if he were still at liberty, the Court notes that the national authorities have made as much allowance as possible for his state of health and his age.
The Court observes, in particular, that some of the matters raised by the applicant (the absence of grab bars in the shower, the short chain on the fanlight and the steep stairs preventing him from reaching the exercise yard) have now been resolved (a seat has been installed in the shower, the chain has been lengthened and the applicant now has permission to walk on the landing). Some of the other inconveniences he mentioned (such as his light being turned on by the warder in the night in order to carry out a check) form part of the constraints imposed on all prisoners for reasons of surveillance and security. It is also clear from the case-file that the applicant keeps up a social life and receives regular visits from his family, friends and lawyers.
In conclusion, having assessed the facts as a whole, the Court finds that, as matters stand at present, the applicant's situation does not attain a sufficient level of severity to come within the scope of Article 3 of the Convention (cf. the Kudła judgment cited above, § 99, and see the Priebke decision cited above).
The Court points out that, if the applicant's health problems, taken together with his extreme old age, were to worsen, French law empowers the national authorities to intervene in various ways. Under Article 729 of the Code of Criminal Procedure, as amended by the Act of 15 June 2000, prisoners may be granted parole if, among other things, they can prove that they "have to undergo treatment". Furthermore, where particularly serious circumstances call for humanitarian measures, the French President may exercise his prerogative of mercy at any time, as provided in Articles 17 and 19 of the Constitution.
It follows that the application is manifestly ill-founded within the meaning of Article 35 § 3 of the Convention.
For these reasons, the Court unanimously
Declares the application inadmissible.
PAPON v. FRANCE DECISION
PAPON v. FRANCE DECISION


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