NIL, not given any of the nanocomposite Rats in the treatment gr

NIL, not given any of the nanocomposite. Rats in the treatment groups received a dose of freshly prepared nanocomposite (100 ml/kg body weight), while rats in the control group received only normal saline daily. Animal’s weights were taken at the start of the dosing (day 0) and weekly thereafter. The animals were observed twice daily for any clinical signs of toxicity and possible mortality during the course of treatment. On day 28 of nanocomposite administration, the animals were sacrificed via exsanguination through cardiac puncture following anaesthesia with ketamine and xylazine.

The brain, liver, spleen, heart and kidney harvested from the rats were weighted individually then examined macroscopically for any abnormality. Coefficients of the brain, liver, spleen, heart and kidney The coefficients of the brain, liver, spleen, heart and kidney, which is the ratio of these organs

to body weight, were calculated after weighing each organ [the ratio of organ (wet weight, mg) to body weight (g)]. Biochemical parameters in serum Blood was collected from rats in each group in a plain 15 mL Falcon tube. It was allowed to stand for about 30 min, before centrifuge at 1,500 rpm, at room temperature. The serum obtained was used for BYL719 concentration the assessment of biochemical parameters. Histopathological

evaluation The animals were subjected to trans-cardiac Branched chain aminotransferase perfusion using 4% paraformaldehyde (PFA). The tissues obtained were processed using the standard procedure and embedded into paraffin blocks, then microsectioned into 5-μm thick and placed onto glass slides. Haematoxylin-eosin (H & E) staining was used on the tissue sections and viewed using optical microscope (FSX-100 Olympus, Olympus Corporation, Shinjiku-ku, Tokyo, Japan). Transmission electron microscope analysis The substantia nigra was dissected from the whole brain perfused and fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.2) for 24 h at room temperature, and was washed twice in 0.1 M phosphate buffer. Then the tissues were post-fixed at room temperature for 4 h in a solution containing 1% osmium tetroxide, 0.8% potassium ferricyanide, 5 mM calcium chloride and 0.1 M cacodylate buffer pH 7.2. The tissues were dehydrated in gradient series of ethanol (20% to 100%) and acetone before embedment in epoxy resin at room temperature. The sections for viewing were made into ultra-thin slices using an ultra-microtome, and they were collected on copper grids and stained with uranyl acetate and lead citrate. The sections were viewed with a Hitachi H-600 transition electron microscope (Chiyoda, Tokyo, Japan) (TEM).

Total RNA of tissue samples and cell lines were isolated by using

Total RNA of tissue samples and cell lines were isolated by using Trizol reagent according to the instruction manual (Invitrogen). Total RNA of leukocytes obtained from 2 ml Roxadustat in vitro of peripheral blood was isolated by using PURESCRIPT RNA Isolation Kit (Gentra systems). RT-PCR Five microgram of the total RNA was reverse transcribed using oligo-dT primer and SuperScript III (Invitrogen) according to the instruction manual. To confirm the expression of Rad18, primer sets, 5′-TTC, ACA, AAA, GGA, AGC, CGC, TG

(forward) and 5′-TTA, CTG, AGG, TCA, TAT, TAT, CTT, C (reverse) were used to amplify 310 bp region of human Rad18 gene. PCR was carried out in a condition of, 3 min at 94°C for initial denaturing, followed by 35 cycles of amplification (94°C for

30 sec, 55°C for 30 sec, and 72°C for 30 sec) using GoTaq (Promega). The amplified products were visualized selleck on 1.2% agarose gel with ethidium bromide. GAPDH in the same samples was also amplified using 25 cycles PCR reaction as the internal control. The primer sets for GAPDH is 5′-TGA, CCA, CAG, TCC, ATG, CCA, TC (forward) and 5′-CCA, CCC, TGT, TGC, TGT, AGC, C (reverse). Fragment Southern Genomic DNA from human breast cancer cell line MCF7 and lung carcinoma cell line PC3 were isolated using TRIZOL according to the instruction manual. MCF7 was used as positive control which was confirmed that this cell line carry wild type Rad18 by RT-PCR direct sequencing (data not shown). Ten microgram of genomic DNA were digested by EcoRI or HindIII, electrophoresed on a 0.8% agarose gel and transferred to a Hybond-NX membrane (Amersham).

Full length cDNA clone of Rad18 was labeled using Psoralen-Biotin nonisotopic labeling kit (BrightStar) and hybridized in PEG-SDS including 100 μg/ml Salmon sperm DNA at 65°C. Detection was done using BioDetect nonisotopic detection kit (BrightStar) according to the instruction manual. Membrane was exposed to X-ray film and developed. RT-PCR SSCP and direct sequencing Benzatropine The primer sets for RT-PCR SSCP are shown in Table 1. Each primer sets were designed to partially overlap the next fragment with the length not more than 200 bp. Ten primer sets cover the whole open reading frame of Rad18 gene and partially, 5′ and 3′ non coding lesion. PCR condition is, 3 min at 94°C for initial denaturing, followed by 35 cycles of amplification (94°C for 30 sec, 55°C for 30 sec, and 72°C for 30 sec). Each sample was denatured 5 min at 95°C and rapidly chilled on ice and loaded into 10% acrylamide gel including 5.4% glycerol for 6 hours at 120V using MiniProtean3 (BioRad) at 4°C. After electrophoreses, gels were stained using Silver Stain Plus Kit according to the instruction manual (BioRad). All samples were screened for the presence of an aberrant band compared with reference sample. Samples with abnormal SSCP bands were directly sequenced by ABI 310. Cycle sequencing was performed using Big-Dye Terminator v3.1 (Applied Biosystems).

PubMed 63 Najbauer J, Johnson BA, Young AL, Aswad DW: Peptides w

PubMed 63. Najbauer J, Johnson BA, Young AL, Aswad DW: Peptides with sequences similar to glycine, arginine-rich motifs in proteins interacting with RNA are efficiently recognized by methyltransferase(s) modifying arginine in numerous proteins. J Biol Chem 1993, 268:10501–10509.PubMed 64. Vance JE, Vance DE: Phospholipid biosynthesis in mammalian cells. Biochem Cell Biol 2004, 82:113–128.PubMedCrossRef 65. Kennedy EP, Weiss SB: The function of cytidine coenzymes

in the biosynthesis of phospholipids. J Biol Chem 1956, 222:193–214.PubMed 66. Vance JE, Steenbergen R: Metabolism and functions of phosphatidylserine. Prog Lipid Res 2005, 44:207–234.PubMedCrossRef 67. Smith TK, Bütikofer P: Lipid metabolism in Trypanosoma brucei . selleck chemicals Mol Biochem Parasitol 2010, 172:66–79.PubMedCrossRef 68. Martin KL, Smith TK: Phosphatidylinositol synthesis is essential in bloodstream form Trypanosoma brucei . Biochem J 2006, 396:287–295.PubMedCrossRef 69. Signorell A, Rauch M, Jelk J, Ferguson MAJ, Bütikofer P: Phosphatidylethanolamine in Trypanosoma brucei is organized in two separate pools and is synthesized exclusively by the Kennedy Pathway. J Biol Chem 2008,

283:23636–23644.PubMedCrossRef 70. Ferguson MAJ: The structure, biosynthesis and functions of glycosylphosphatidylinositol anchors, and the contributions of trypanosome research. J Cell Science 1999, 112:2799–2809.PubMed 71. Martin KL, Smith TK: The glycosylphosphatidylinositol (GPI) biosynthetic pathway of bloodstream form Trypanosoma brucei is dependent on the de novo synthesis of inositol. Mol Microbiol 2006, 61:89–105.PubMedCrossRef 72. Menon AK, Eppinger M, Mayor S, Schwarz RT: Phosphatidylethanolamine is the donor selleck compound of the terminal phosphoethanolamine group in trypanosome glycosylphosphatidylinositols. EMBO J 1993, 12:1907–1914.PubMed 73. Gibellini F, Hunter WN, Smith TK: The ethanolamine branch of the Kennedy pathway is essential in the bloodstream form of Trypanosoma brucei . Mol Microbiol 2009, 73:826–843.PubMedCrossRef 74. Brun R, Schonenberg M: Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Acta Clomifene Trop 1979, 36:289–292.PubMed

75. Gietz D, St-Jean A, Woods RA, Schiestl RH: Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 1992, 20:1425.PubMedCrossRef 76. Hayman ML, Miller MM, Chandler DM, Goulah CC, Read LK: The trypanosome homolog of human p32 interacts with RBP16 and stimulates its gRNA binding activity. Nucleic Acids Res 2001, 29:5216–5225.PubMedCrossRef 77. Zeiner GM, Sturm NR, Campbell DA: Exportin 1 mediates nuclear export of the kinetoplastid spliced leader RNA. Eukaryot Cell 2003, 2:222–230.PubMedCrossRef 78. Chapman AB, Agabian N: Trypanosoma brucei RNA polymerase II is phosphorylated in the absence of carboxyl-terminal domain heptapeptide repeats. J Biol Chem 1994,269(7):4754–4760.PubMed Competing interest The authors declare that they have no competing interest.

15 pKD46 100 5 2 0 26 pACBSR 100 2 8 1 5 0 pRW50 100 1 2 1 1 0 pU

15 pKD46 100 5 2 0.26 pACBSR 100 2.8 1.5 0 pRW50 100 1.2 1.1 0 pUC18PCR 100 57 15 1 Since the Datsenko and Wanner system

relies upon the introduction of PCR generated DNA into cells and not plasmids that have been isolated from selleck chemical an E. coli K-12 strain, we re-examined the DNA uptake efficiencies of the strains when transformed with a PCR generated version of the plasmid, pUC18. We reasoned that plasmids isolated from a K-12 strain may be subject to host restriction-modification systems in pathogenic strains, hence, using a PCR-generated pUC18 derivative would not only more closely resemble the conditions used by Datsenko and Wanner, but also allow us to monitor the transformation efficiencies by means of the acquired ampicillin resistance due to pUC18 plasmid uptake. Thus, we amplified pUC18 by PCR and then incubated the reaction with DpnI, which specifically digested the methylated template plasmid and not the PCR generated MK-1775 product. The PCR generated pUC18 plasmid (pUC18PCR) was then transformed into MG1655, CFT073, O157:H7 Sakai and O42 by electroporation. The results (table 1) show that the transformation frequency of the pathogenic strains by pUC18PCR was slightly improved when compared with MG1655, although the overall transformation frequency remains far lower than MG1655. The overall number of MG1655

colonies identified after transformation with pUC18 or pUC18PCR was comparable. Thus, the electroporation step is likely to be the primary reason for the poor efficiency of this system in pathogenic E. coli strains. This shortcoming was alleviated somewhat by Murphy and Campellone

Florfenicol [15] who developed an improved electroporation based protocol for recombineering in E. coli EHEC and EPEC strains. However, we have had mixed success using this protocol, particularly when recombineering in EAEC and UPEC strains, where no increase in recombination frequency was observed. B. Two-plasmid recombineering The two plasmid gene-gorging method described by Herring and co-workers [4] has an immediate advantage for recombineering in pathogenic strains since the method does not rely upon efficient electroporation as a means of introducing target DNA into the cell. Instead, the target DNA is flanked by recognition sites for the meganuclease I-SceI on a donor plasmid that is transformed into cells along with the recombineering plasmid, pACBSR, which carries I-SceI and the λ-Red genes whose expression is controlled by an arabinose inducible promoter. Induction of I-SceI results in donor plasmid cleavage, generating the linear dsDNA target, which is a substrate for λ-Red gene products. Herring and co-workers disrupted chromosomal genes by introducing amber mutations, using long regions of homology to the chromosome and reported that the recombination frequency for gene gorging was between 1-15%.

The results of the tests for examining intragenic recombination (

The results of the tests for examining intragenic recombination (recombination within the sequence of a gene) are summarised in Table  2. For each test the number of loci that were positive for recombination is recorded. For RDP at least two of the individual tests in the suite had to Selleck RAD001 be positive in order for the locus to be

scored positive overall. Table 2 Number of loci positive for recombination by the Sawyer’s run test and RDP suite   Sawyer’s run test RDP tests Staphylococcus aureus (Clonal) 0 loci 1 locus Streptococcus pneumoniae (Intermediate) 3 loci 4 loci Neisseria menigitidis (Panmictic) 7 loci 6 loci Legionella pneumophila 1 locus 2 loci Both the Sawyer’s run test and RDP show L. pneumophila has an intermediate rate of

intragenic recombination when compared with other bacterial species. Overall the collected evidence from this and several previous studies [12–14, 16, 17, 23] strongly suggest that L. pneumophila is not a purely clonal organism but also undergoes significant recombination. The results presented here suggest that L. pneumophila retains evidence for a clonal vertical inheritance of genetic material whilst also demonstrating strong evidence of recombination by horizontal transfer of genetic loci. Although there was some evidence for recombination within the SBT genes, the frequency was low and this indicates selleck chemical that new alleles are most likely to be generated by point mutations 2-hydroxyphytanoyl-CoA lyase rather than recombination. The signal from vertical inheritance of genetic material through clonal lineages is still evident when examining the genetic information contained from seven L. pneumophila loci. However it is also clear that recombination happens often enough so that it is a significant force in shaping the population structure. This does not alter the utility of SBT as a means to discriminate between isolates of L. pneumophila, particularly for outbreak investigation, since the results indicate that it is far from being a panmictic organism. Although we

cannot infer a rate of recombination from this study, the relatively low frequency of recombination suggests that recombination would be unlikely to take place in the timescale of an outbreak and therefore the ST of isolates involved in an outbreak is also unlikely to change. Sequence Based Typing analysis: Clustering Since the ultimate aim of this work was to find a practical way to cluster L. pneumophila isolates, a method of determining which clustering method resulted in the most accurate sub-groups was required. Given that the recombination analysis above indicates that clonal vertical inheritance plays a major role in the evolution of L. pneumophila, a phylogenetic tree based on the genetic distance between the concatenated sequences from the SBT loci will provide an approximate representation of the evolutionary history.

It has been reported that the release of cyto c appears to

It has been reported that the release of cyto c appears to

be dependent on the induction of mitochondrial permeability transition, which is associated with a decrease in Δφm; therefore, the loss of Δφm and the release of apoptogenic factors, such as cyto c, from the mitochondria into the cytosol are associated with apoptosis induced by chemotherapeutic drugs[25–27]. In the present study, loss of Δφm and release of Cyto c were observed in NCTD-treated cells, resulting in caspase-9 and caspase-3 activation and PARP cleavage and, finally, apoptosis. Moreover, the loss of Δφm may, in fact, be a consequence of massive cytochrome c release from the mitochondria. Thus, a mitochondrial damage-dependent pathway may be involved in NCTD-induced apoptosis in HepG2 cells. Some studies have reported that ROS act as secondary messengers in apoptosis induced by anti-cancer and chemopreventive agents[28, 29]. The generation of ROS can cause the loss of Δφm, and induce apoptosis by releasing pro-apoptotic proteins such as AIF and Cyto c from mitochondria to the cytosol.The generation of ROS may contribute to mitochondrial

damage and lead to cell death by acting as an apoptotic signaling molecule[30, 31]. To reveal if NCTD influenced the level of ROS, we stained drug treated cells with DCFH-DA. We found that, in addition to its effect on Δφm, NCTD caused an increase in ROS production in HepG2 cells. The NCTD -induced increase in ROS and antiproliferation in HepG2 cells are apparently dependent on ROS generation, because the NCTD -induced increase in ROS can be abolished or MG-132 solubility dmso attenuated by antioxidants, such as NAC. In addition, we found that NCTD -induced antiproliferation in HepG2 cells was also abolished by the antioxidant NAC. Conclusions In conclusion, our data indicate that NCTD induced apoptosis in HepG2 cells via ROS generation and mitochondrial pathway (Figure 7)[32]. These findings suggest that NCTD

may one day be used in the prevention and treatment of cancer. Figure 7 A proposed model showing the mechanism of NCTD anti-proliferative and apoptosis effects in HepG2 cells. ROS, reactive oxygen species; PARP, poly (ADP ribose)polymerase; Δφm, mitochondrial membrane potential; Sulfite dehydrogenase Apaf-1, apoptotic protease activating factor-1. Acknowledgements We thank Yan Wan, Department of Immunology, Wuhan University, for exceptional technical assistance in flow cytometry analysis. References 1. El-Serag HB, Rudolph KL: Hepatocellular carcinoma:epidemiology and molecular carcinogenesis. Gastroenterology 2007, 132: 2557–2576.PubMedCrossRef 2. Wang GS: Medical uses of mylabris in ancient China and recent studies. J Ethnopharmacol 1989, 26: 147–162.PubMedCrossRef 3. Peng F, Wei YQ, Tian L, Yang L, Zhao X, Lu Y: Induction of apoptosis by norcantharidin in human colorectal carcinoma cell lines: involvement of the CD95 receptor/ligand. J Cancer Res Clin Oncol 2002, 128: 223–230.PubMedCrossRef 4.

Two isolates (H063920004 and H091960009) were sequenced with diff

Two isolates (H063920004 and H091960009) were sequenced with different technologies. H063920004 with Illumina paired end, Illumina mate-paired and Roche 454, H091960009 with Illumina paired end and Illumina mate-paired. There were

no SNP differences between the sequences of these replicate samples demonstrating that the protocol used for calling SNP variants is both robust and consistent. There were three isolates of ST47 (labelled ST47, LP-617 and Lorraine), two from the UK and one from France, each isolated in a different year between 2003 and 2006. These differed by just four SNPs. Two ST42 isolates, from the UK and USA (labelled check details ST42 and Wadsworth), were isolated 20 years apart and only exhibited 20 SNP differences. In contrast two ST1 isolates, a representative of the ‘Paris’ strain and a UK strain sequenced as part of another study, were isolated within 2 years of each other yet these exhibited 280 SNP differences. These results show that lineages of L. pneumophila contain differing levels of observable diversity. There are several evolutionary scenarios that could be postulated Rapamycin cost as explanations for these observed differences. A lineage that occupies a niche where there is strong purifying selection will be less diverse. Conversely a lineage that is the result of rapid expansion

within a previously unoccupied niche will tend to be more diverse. One likely scenario is that ST1 is a successful clonal lineage that emerged before the ST47 lineage and therefore has had more time to diversify by genetic drift. It is also possible that each lineage of L. pneumophila will be subject to differing selection pressures when infecting a human host, even though this is effectively an evolutionary dead-end. One possible scenario is that the majority of ST1 strains

and a limited number of sub-lineages of ST47 cause disease in humans. If this is the case then a likely explanation is that the common ancestor of the ST1 lineage was able to infect the human species and the ancestor of the ST47 lineage did not replicate effectively in a human host. Subsequently a minority of descendents of the ST47 lineage have acquired click here the ability (through mutation, gene loss or acquisition) to cause human infection. Differentiating between these putative evolutionary scenarios will be difficult and will require a greater understanding of the effects of diversity within the lineages of L. pneumophila sampled from the environment and human infections. When examining the output from the Splits Tree analysis, the more splits observed, the more recombination or HGT is likely to have taken place. The majority of clades in the tree show a branching network structure suggestive of frequent recombination. The Phi test for recombination as implemented in SplitsTree also showed evidence for recombination (p = 0.0). The exceptions are the clade(s) containing ST136/154 and ST707.

For a flat surface having an AR overlayer, using Fresnel’s reflec

For a flat surface having an AR overlayer, using Fresnel’s reflection formula, we measured the

reflectance at different wavelengths. It is observed that with varying film thickness, the position of the reflection minima shifts, while a change in the refractive index modifies buy Torin 1 the amount of surface reflectance [25]. Although similar trends are quite evident, the experimentally observed average surface reflectance turns out to be much lower over the spectral range under consideration. In order to explain these results, let us first try to understand the role of the Si template which is practically an ensemble of ion beam-fabricated self-organized conical nanofacets at the top of the Si substrate. It is known that grating on any surface can be used to achieve arbitrary refractive index if the geometry of the grating structures can be tuned. For instance, if we consider a binary grating, its effective refractive index, n eff, can be expressed as n eff = (n 1 - 1)DC + 1, where n 1 is the refractive index of the grating and DC is the duty cycle and is defined as the ratio of the grating

line width to the grating period [26]. If the surrounding medium is taken as air and the grating is of the same material as the substrate, the optimized duty cycle (to meet the AR criterion) can be expressed as where n 2 is the refractive index GDC-0199 cell line of the substrate [26]. Such binary gratings are expected to exhibit the AR property over a very narrow spectral range. This range can be broadened by continuous tuning of the refractive index (n eff) between the two surrounding media. This would essentially mean

a continuous change in DC along the depth (from the apex towards the base of the facets) of the grating lines, which is possible to be achieved by having tapered/conical gratings. When the grating and the substrate materials are the same, the matching of refractive index at the substrate interfaces can exhibit highly Celecoxib improved AR property [27]. This explains the enhanced AR performance observed here for the faceted Si surface formed on the Si substrate. Following the same argument, further improved AR performance is expected due to the conformal growth of an AZO overlayer on nanofaceted Si template. Indeed, the experimental findings confirm the same where increasing AZO thickness leads to a systematic red shift in the reflection minima. However, such small variations in the thickness may not be sufficient to cause any significant difference in depth-dependent change of the effective refractive index for the AZO-coated faceted Si template which corroborates well with the experimentally measured reflectance minima values.

Expression of

FHL is maximal under fermentative condition

Expression of

FHL is maximal under fermentative conditions in the absence of exogenous electron acceptors and is absolutely dependent on formate [13]. Hyd-3 is considered a labile hydrogenase that has so far proven recalcitrant to isolation in an active form [14]. The labile molybdenum- and selenium-dependent formate dehydrogenase-H (Fdh-H) Enzalutamide in vivo is also associated with the FHL complex [15]. Fdh-H represents one of the three formate dehydrogenase enzymes in E. coli (Fdh-H, Fdh-O, and Fdh-N) [16]. Fdh-O and Fdh-N are membrane-bound and periplasmically-oriented respiratory enzymes that couple formate oxidation to quinone reduction and thus contribute directly to energy Selleck NVP-LDE225 conservation. Several methods have been described for visualizing the redox activity of hydrogenases. Most commonly, low-potential artificial redox-active viologen dyes such as methyl viologen (MV) and benzyl viologen (BV)

have been used [17, 18]. All three E. coli hydrogenases can couple H2 oxidation to BV reduction in vitro and when extracts from fermentatively-grown cells are assayed Hyd-3 can contribute over 90% to the total activity [19, 20]. While Hyd-1- and Hyd-2-catalysed BV reduction can be readily visualised and the enzymes distinguished by use of an in-gel assay [18], Hyd-3 activity has so far proved recalcitrant to zymographic identification and this had been thought to be due to the instability of the large FHL complex (see [1]). Moreover, the large respiratory Fdh-N and Fdh-O enzyme complexes also contribute some background staining due to their inherent H2:BV oxidoreductase activities, thus making any assessment of a Hyd-3 associated activity potentially problematic [21]. Alternative hydrogenase assays have been developed isometheptene for other biological systems. For example, the oxygen-tolerant hydrogenases from Ralstonia eutropha H16 can be visualized with phenazine methosulfate (PMS)/nitroblue tetrazolium (NBT) [22] or PMS/triphenyl tetrazolium chloride (TTC) [23] combinations

of redox dyes. Methylene blue has also been used extensively in hydrogenase research [24]. However, the use of alternative redox-active electron acceptors has not really been extensively explored for the hydrogenases of E. coli. The aim of this study, therefore, was to investigate the differential activities of the E. coli hydrogenases with a view to making it possible to distinguish all enzymes synthesized under anaerobic growth conditions. We describe here conditions that allow the unequivocal visualization of all three, membrane-associated, anaerobically inducible hydrogenase enzyme complexes. Results Identification of Hyd-3 activity through an in-gel assay Hyd-1 and Hyd-2 are readily visualized after gel electrophoresis under non-denaturing conditions in a high-pH buffering system [18–20].

e , female-headed households, households lacking able-bodied men

e., female-headed households, households lacking able-bodied men aged 19–35 years, households with many dependants and households with many sick family members. In order to implement this in practice we therefore suggest [in contrast to the national adaptation policies proposed by the governments in Tanzania and Kenya but in agreement with IFAD recommendations (2011)] a gender-informed and tri-partite integrative policy strategy with focus on: (1) financial and infrastructural support to scale up adoption of locally produced and

affordable technologies and innovations; (2) education and extension services targeting and promoting a shift towards sustainable agricultural intensification; and

(3) capacity building and social learning initiatives to encourage the integration of “marginalized” climate vulnerable groups into collaborative projects and collective action groups to reduce labor burdens and CHIR-99021 ic50 diversify activities and income earning possibilities. In so doing, three important livelihood domains may be promoted and developed: the capability to farm collectively; the means to increase household buffers; and the empowerment of individual agency to enable planning for the uncertainties ahead. Acknowledgments The authors would like to thank the Swedish International Development and Cooperation Agency (Sida) for financial support of the research project and three anonymous reviewers for insightful comments on the article. We also wish to thank all the participating stakeholders in the Kisumu workshop and SCC-VI Kisumu for arranging and

co-hosting the event. Finally, our gratitude goes to the farmers who engaged Dimethyl sulfoxide so willingly in the participatory exercises. Without you this research would not have been possible. Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. References Adger WN (2003) Social capital, collective action, and adaptation to climate change. Econ Geogr 79(4):387–404CrossRef Adger WN (2006) Vulnerability. Global Environ Change 16(3):268–281CrossRef Andersson E, Gabrielsson S (2012) Because of poverty we had to come together—collective action as a pathway to improved food security in rural Kenya and Uganda. J Int Agric Sustain 10(3):245–262 Barrett CB (2008) Poverty traps and resource dynamics in smallholder agrarian systems. In: Dellink RB and Ruijs A (eds) Economics of poverty, environment and natural-resource use.