Such a result suggests that the markers

do not share the same genealogy, likely due to extensive recombination or re-assortment Ro-3306 breaking down linkage between markers. The diversity of E. histolytica genome raises a concern in regard to later analysis as it raises the possibility that a rapid rate of evolution may drive any observed differences between E. histolytica genotypes in samples isolated in regions separated even by relatively small geographical distances. Figure 3 Lack of consistent patterns of descent among SNP markers from Bangladeshi E. histolytica isolates suggests they segregate independently. Consensus phylogeny inferred from 100 bootstrap replicates of polymorphic SNP markers, constructed using the MEGA 5 program and the Maximum Likelihood method based on the Tamura-Nei model and using the sequences

shown in Additional file 1: Table 8 [42]. Branches produced in fewer than 50% of the bootstrap phylogenies were collapsed. Sequences from stool have the suffix s; culture c; monthly survey stools begin with MS or CMS, diarrheal DS or CDS, amebic liver abscess samples RUF. The effect of adaptation of to in vitro culture on SNP allele frequencies To examine the potential effect of adaption Tucidinostat research buy to in vitro culture on the frequency of SNP alleles, and therefore how well transiently or long established cultured trophozoites represent the parasite population, SNP allele frequencies were compared Tangeritin between parasites genotyped directly from stool samples and those from cultured trophozoites

(Additional file 1: Table S10). In cultures originating from asymptomatic isolates five linked Non-Reference SNPs at the LCAT EHI_065250/XM_647310.1 locus were detected in 80% of the strains, these same SNPs occurred in only 16% of the E. histolytica positive stool samples from asymptomatic hosts (Figure 4). This suggests that during establishment of E. histolytica cultures a strong selection pressure was exerted on sequence in linkage with the LCAT EHI_065250 gene. This could either cause growth failure of the strains with the Reference allele or the outgrowth of a minority genotype in mixed infections (previous studies using the short tandem repeats have MK-8931 indicated that mixed infections are rare however this possibility cannot be discounted [24]). Figure 4 Amebic culture effect on the EHI_065250 Entamoeba genotype. Distribution of the EHI_065250 SNP at the 10296 location in field isolates or cultured strains established from asymptomatic disease (p = 0.0166). The distribution of the individual SNPs, which were either Reference (Ref), Non-Reference (Non-Ref) or heterologous was shown on the x-axis. The number of samples of with this genotype isolated from patients with asymptomatic disease was shown on the y-axis.

NOR is pinkish in color. After 4-d cultures, the control plate was pink in color, while no color was observed in the plate with 40 mg/mL D-glucal. Spectrophotometric analyses showed that NOR productions were significantly inhibited by selleck inhibitor D-glucal at concentrations of 10 mg/mL or higher (Figure 4C). These results suggest that D-glucal inhibits the

AF biosynthesis pathway prior to the production of NOR. D-glucal inhibited expression of AF biosynthetic genes, but promoted expression of kojic acid biosynthetic genes To examine the effect of D-glucal on AF biosynthesis at the transcriptional level, we analyzed expression of several genes in the AF biosynthetic gene cluster in A. flavus A 3.2890 by qRT-PCR and observed that, in the presence of 40 mg/mL

D-glucal, no significant change was detected for aflR [a Zn (II)2 Cys6 IAP inhibitor transcription factor], while a 28% reduction was observed for aflS (a co-activator, Figure 5A). In addition, expression levels of all seven genes encoding AF biosynthetic enzymes tested, aflC (polyketide synthase), aflD (oxidoreductase), aflM (dehydrogenase), aflO (O-methyltransferase B), aflP (O-methyltransferase A), aflU (P450 monooxygenase) and nadA (a cytosolic enzyme converting AFB1 to AFG1), were decreased significantly (Figure 5A). Among these, aflC encodes an upstream enzyme in AF biosynthesis pathway, ERK inhibitor acting before NOR production to synthesize the polyketide backbone [21], while nadA encodes the most downstream enzyme, converting AFB1 to AFG1 [22, 23]. Figure 5 Expression analyses of genes for AF and kojic acid production and sugar utilization. (A) qRT-PCR analyses of expression of 9 AF biosynthetic genes (aflR, aflS, aflC, aflD, aflM, aflP, aflO, aflU, and nadA) and 3 sugar utilization genes (hxtA, glcA and sugR) in mycelia grown

with or without 40 mg/mL D-glucal for 3 d, The relative expression levels were quantified through comparison with the expression level of β-tubulin. Data are presented as means ± S.D. (n = 3). (B) Expression of 3 kojic acid biosynthetic genes (kojA, kojR, kojT) by qRT-PCR mafosfamide in mycelia grown with or without 40 mg/mL D-glucal for 3 d. The relative expression levels were quantified through comparison with the expression level of β-tubulin. Data are presented as means ± S.D. (n = 3). We then examined if the expression levels of genes in the sugar utilization gene cluster were changed when cultured in media containing D-glucal. Of three genes tested, sugR (transcriptional regulator), hxtA (sugar transport), and glcA (glycosylation), none showed significant changes in expression (Figure 5A).

Acta Physiol Scand 1973,89(3):374–383.CrossRefPubMed 22. Norman B, Sollevi A, Jansson E: Increased IMP content in glycogen-depleted muscle fibres during submaximal exercise in man. Acta Physiol Scand 1988,133(1):97–100.CrossRefPubMed 23. Ivy JL: Dietary strategies to promote glycogen synthesis after exercise. Can J Appl Physiol 2001, 26:S236-S245.PubMed 24. Wolfe RR: Effects of amino find more acid

intake on anabolic processes. Can J Appl Physiol 2001, 26:S220-S227.PubMed 25. Allen DG, Lannergren J, Westerblad H: Muscle cell function during prolonged activity: cellular mechanisms of fatigue. Exp Physiol 1995,80(4):497–527.PubMed 26. O’Leary DD, Hope K, Sale DG: Posttetanic potentiation of human dorsiflexors. J Appl Physiol 1997,83(6):2131–2138.PubMed 27. Fernstrom JD: Branched-chain amino acids and brain function. J Nutr 2005,135(6 Suppl):s1539-s1546. 28. Markus CR, Olivier B, de Haan EH: Whey protein rich in alpha-lactalbumin increases the ratio of plasma tryptophan to the sum of the other large neutral amino acids and improves cognitive performance in stress-vulnerable subjects. Am J Clin Nutr 2002,75(6):1051–1056.PubMed 29. Davis JM, Bailey SP: Possible mechanisms of central nervous system fatigue during

exercise. Med Sci Screening Library Sports Exerc 1997,29(1):45–57.PubMed 30. IOM: Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington: The national academies press; 2002. 31. Trabulsi J, Schoeller DA: Evaluation of dietary assessment instruments

against doubly labeled water, a biomarker of habitual energy intake. Am J Physiol Endocrinol Metab 2001,281(5):E891–899.PubMed 32. Westerterp KR, Donkers JH, Fredrix EW, Boekhoudt P: Energy intake, physical activity and body weight: a simulation model. Br J Nutr 1995,73(3):337–347.CrossRefPubMed 33. Valentine RJ, Saunders MJ, Todd MK, St Laurent TG: Influence of carbohydrate-protein beverage on cycling endurance and indices of muscle disruption. Int J Edoxaban Sport Nutr Exerc Metab 2008,18(4):363–378.PubMed 34. Tipton KD, Wolfe RR: Protein and amino acids for athletes. J Sports Sci 2004,22(1):65–79.CrossRefPubMed Competing interests The authors declare that they have no competing interests. Authors’ contributions SB, JB, JF and MW all contributed to the study design. SB and NW recruited participants and conducted all data collection. SB undertook analysis of all the data. SB and MW both interpreted the data. All authors reviewed and approved the final manuscript.”
“Background DL-α-hydroxy-isocaproic acid (HICA), also known as leucic acid or DL-2-hydroxy-4-methylvaleric acid, is an end product of leucine metabolism in human tissues such as muscle and connective tissue [1, 2]. Some Belinostat supplier foodstuffs produced by fermentation, e.g. certain cheeses, wines and soy sauce contain HICA [3–7].

05% Congo Red (w/v). SD1 in vitro find more samples were prepared by inoculating a single colony into Luria-Bertani (LB) medium grown to stationary phase at 37°C with agitation. The bacteria were harvested by centrifugation and washed twice with ice-cold PBS (6,000 × g, 15 min) at 4°C. The inoculum for in vivo experiments

was prepared by growing a typical SD1 colony selected from a TSA plate in LB medium overnight. Gnotobiotic piglets used for the animal experiments were delivered by Caesarian section at Tufts University Cummings School of Veterinary Medicine. Of several animals inoculated with SD1, three piglets were chosen for isolation of SD1 bacterial https://www.selleckchem.com/products/a-769662.html cells from the intestine in this comparative study. One of the piglets inoculated with 1 × 108 SD1 cells developed diarrhea 24 h later and was euthanized 4 d later when the gut contents SAHA HDAC concentration were collected for bacterial purification. Another piglet inoculated with 5 × 108 SD1 cells developed diarrhea within 18 h and was euthanized 3 d post-inoculation. A third piglet inoculated with 5 × 109 SD1 cells developed diarrhea within 20 h and the

gut contents collected 2 d post-inoculation. SD1 bacterial cells were isolated from the gut contents as described previously [15]. Briefly, the gut contents from cecum and colon were pooled and transferred to sterile histological cups placed on ice, suspended in ice-cold PBS at 4°C and pelleted at 5,000 × g. After resuspension of the pellet in 65% isotonic Percoll solution and centrifugation at 14,500 × g, the bacterial layer near the bottom was collected using a 3-5 ml syringe with needle. The bacteria were washed twice with ice-cold PBS at 4°C and processed for proteomic analysis. Lysis of S. dysenteriae cells and trypsin digestion of extracted proteins After the PBS wash steps, bacterial cell pellets from in vitro or in vivo culture conditions were re-suspended in a hypotonic lysis buffer composed of 25 mM Tris-HCl (pH 7.8) with 150 μg/mL lysozyme, 0.05% Triton X-100, 5 mM EDTA, protease inhibitors (1 mM benzamidine and AEBSF) for 30 Olopatadine min at

room temperature (RT) with gentle agitation. The samples were then placed at -80°C until further processing. For nucleic acid digestion, bacterial samples suspended in the lysis buffer were thawed and gently agitated for 1 h at RT after the addition of DNase I, RNase and leupeptin (10 μg/mL each) and 20 mM MgCl2. Cell lysates were centrifuged at 16,000 × g for 30 min at 4°C, and the supernatants containing bacterial cell lysate proteins were recovered. Following cell lysis, the extracted bacterial proteins were precipitated in six volumes of ice-cold acetone at -20°C for at least 1 h. Acetone-precipitated proteins were recovered as a pellet after centrifugation at 5,000 × g for 10 min. The protein pellet was resuspended in 0.1 M TAB buffer, pH 8.5, and the total protein concentration measured using the BCA assay. Proteins were denatured in 0.

Collection of Emricasan sputum samples and microbial culture Spontaneously expectorated sputum samples were collected from consecutive outpatients AP26113 within a cohort of adult NCFBr patients. The samples were washed with phosphate-buffered saline to remove any contamination from oral flora [12]. Each sample was homogenised with Sputasol (Oxoid) and divided into two aliquots, one for subsequent DNA extraction

and one for immediate culture, performed in accordance with national standard methods in an accredited UK clinical laboratory. Briefly, 10 μL aliquots of homogenised sputum were cultured onto Columbia blood agar and Chocolate agar plus bacitracin. The sample was subsequently diluted 1/100 in sterile saline (0.85%) and 10 μL of this was cultured onto

chocolate agar and incubated in air plus 5% carbon dioxide (37°C, 48 h). Isolates were identified by matrix assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry (Bruker Daltonics) and, where necessary, appropriate API kits (bioMérieux) [29]. Information, from up to 10 years previously on prior P. aeruginosa status, was collected (Additional file 1: Table S1). Persistent infection was defined as isolation ofa taxa from previous sputum samples Doramapimod with a minimum requirement of having been cultured on two or more occasions [2] based upon current and prior sputa culture data. Intermittent colonisation was defined as isolation of taxa from a patient’s sputa preceded or followed by sputa that was culture negative. DNA was extracted from 0.5 ml of each sputum sample using the MoBio Ultraclean Microbial DNA isolation kit (MoBio, CA, USA) according to the manufacturer’s protocol. A

negative control where template DNA was replaced with sterile distilled water was prepared with the same reagents. Extracted DNA was quantified with a NanoDrop 1000 Spectrophotometer (Thermo Scientific). 454 Pyrosequencing From standardised concentrations of template DNA a Rebamipide portion of 16S rRNA gene (position 341 to 907; Escherichia coli numbering) was amplified using the primer set 341 F and 907R [30]. DNA sequencing was performed using the 454 GS FLX Titanium Sequencing System (Roche, IN, USA) by the Research and Testing Laboratory (RTL, TX, USA) using previously described methods [31]. The raw sequencing reads were quality filtered in QIIME 1.6.0 [32] using the split-library.py script. Remaining high quality sequences were clustered into operational taxonomic units (OTUs) at 97% similarity using UCLUST [33]. Representative sequences for each OTU were aligned using PyNAST [34] and taxonomic identities were assigned using RDP-classifier (version 2.2) [35] with 50% as confidence value threshold. Detection of potentially chimeric sequences was performed using ChimeraSlayer [36] and chimeric sequences were removed from downstream analysis prior to tree building using FastTree [37].

We isolated a single protein, IsaB. Subsequently, we found that IsaB did not play a role in regulation of ica expression, was not localized to the cytoplasm where it could potentially play a regulatory role but

rather was secreted and partially selleck compound associated with the bacterial cell surface, and bound to RNA, ssDNA, and dsDNA with no apparent sequence specificity. Because a number of studies have shown a role for extracellular DNA in biofilm formation, we hypothesized that ON-01910 the extracellular DNA-binding protein IsaB could play a role in this process [18–21]. However, we found that IsaB did not contribute to biofilm formation under a variety of conditions. This study is the first to assign a function to the putative virulence factor IsaB. The physiologic role of binding extracellular nucleic acids is still unclear. Results Isolation of IsaB by RNA Affinity Chromatography We hypothesized that an RNA-binding protein could regulate ica expression at the post-transcriptional level through binding to the 5′-untranslated region (5′-UTR). To isolate factors that bound to the 5′-UTR, we designed an RNA Affinity Chromatography assay using a biotinylated chimeric oligonucleotide (WTUTR-c) based on the sequence upstream Mocetinostat in vivo from the ica locus as shown in Table 1. The 3-nt at the Anacetrapib beginning and end were

synthesized as deoxyribonucleotides to protect the oligo from exoribonuleases, and the remaining 40-nt were ribonucleotides. The chimeric

oligo was immobilized on streptavidin-coated magnetic particles, which were used to isolate proteins from whole cell lysates of S. aureus strain MN8. A single 19.5 kDa protein was detectable by Coomassie staining (data not shown), and was identified by Mass Spectral analysis as the immunodominant surface antigen B (IsaB). Table 1 Oligonucleotides used in this study are shown Oligo name Sequence WTUTR-c 5′-BIOTIN-TGCaauuacaaauauuuccguuuaauuauaacaacaaucuauuGCA-3′       IsaBIntein 5′-GGGCATATGAATAAAACCAGTAAAGTTTGTGTAGC-3′         IsaBInteinREV 5′-GGTTGCTCTTCCGCAACCTTTACTTGTTTTGTATGGTGTATGTCC-3′ isaBDELFWD 5′-GGATCCCGGATTTAGGCAATTCTTTTAATGC-3′              isaBDELREV 5′-GGATCCCATTAGAACTAATGTGCTTTGATGG-3′             isaBXhoFWD 5′-GGGCATATGGTTTGTGTAGCAGCAACATTAGC-3′            isaBXhoREV 5′-GGGCTCGAGCGAAGTAACAGTTGGACATACACC-3′           icaUTR6 5′-GUUUAAUUAUAACAACAAUCUAUUGCA-3′                BioticaPRO 5′-BIOTIN-ATTGVGTTATCAATAATCTTA-3′               IcaRcloneFWD 5′-GGTGGGATCCTTGAAGGATAAGATTA-3′            WTUTR(RNA) 5′-Biot-tegugcaauuacaaauauuuccguuuaauuauaacaacaaucuauuGCA-3′        Deoxyribonucleotides are shown in capital letters and ribonucleotides are shown in lower case.

However, this mutation has been described earlier as being specific for the Haarlem genotype and is not

associated with resistance to EMB [16]. As mentioned above, other so far unknown resistance mediating mechanisms are probably responsible for the resistance phenotype in these four strains. Mutations or insertions in the pncA gene are known to mediate PZA resistance [42, CAL-101 order 43], as observed in our study. No hotspot region has been determined, since polymorphisms occur throughout the complete gene. However, according to our data some specific mutations do obviously not mediate resistance that is detectable by applying standard critical concentrations. In the panel of strains analyzed, two susceptible strains carry a SNP at codon 47 and one displays a mutation at codon 96. PZA-MIC determination for these strains revealed slightly elevated values for the strains carrying I-BET-762 supplier the mutation at codon 47 (25.0 μg/ml) compared to the H37Rv control. In a recent study it has been shown that the site of the mutation is leading to varying efficiencies of the mutated pyrazinamidase mediating a wide range of resistance levels from low to high [44]. As the mutation at codon 47 has previously been described

by Juréen and co-workers [42] in PZA resistant strains, further investigations are necessary to determine if additional mutations in other parts of the genome might be responsible for the observed low-level resistance in the strains analyzed in this study. Out of all PZA resistant strains three carried the pncA wild type sequence. This indicates that further Niclosamide mutations in as yet unidentified genes are also important for mediating PZA resistance. Conclusions Although resistance mechanisms to INH and RIF are well understood, unknown resistance determining regions and resistance mediating mechanisms appear to play an important role for SM, EMB and PZA, where we observed

a relatively low sensitivity for selleck chemicals llc detection of resistance by analysis of common genes. Therefore, it is essential to gather information on further mechanisms leading to drug resistant MTBC strains. For the design and implementation of molecular resistance assays it is fundamental to consider strain diversity with respect to resistance mutations in a given geographical setting. Finally, it should be noted that not all variations in well described resistance genes are related to the development of high-level resistance, a finding arguing for a very careful interpretation of molecular resistance assays. Acknowledgments We thank I. Razio, P. Vock, T. Ubben and L. Dost, Borstel, Germany, for excellent technical assistance. Parts of this work have been supported by the European Union TM-REST (FP7-202145) and the TB-PAN-NET (FP7-223681) projects. Electronic supplementary material Additional file 1: PCR primers and conditions used for amplification and sequencing.

Neuromolecular Med 2002,

2:215–231.CrossRef 63. Du L, Zhang X, Han YY, Burke NA, Kochanek PM, Watkins SC, Graham SH, Carcillo JA, Szabó C, Clark RS: Intramitochondrial poly (ADP-ribosylation) contributes to NAD+ depletion and cell death induced by oxidative stress. J Biol Chem 2003, 278:18426–18433.CrossRef 64. Zeng J, Yang GY, Ying W, Kelly M, Hirai K, James TL, Swanson RA, Litt L: Pyruvate improves recovery after PARP-1-associated energy failure induced by oxidative stress in neonatal rat cerebrocortical slices. J Cereb Blood Flow Metab 2007, 27:304–315.CrossRef 65. Araki T, Sasaki Y, see more Milbrandt J: Increased nuclear NAD biosynthesis and SIRT1 activation AC220 concentration prevent axonal degeneration. Science 2004, 305:1010–1013.CrossRef 66. Wang J, Zhai Q, Chen Y, Lin E, Gu W, McBurney

MW, He Z: A local mechanism mediates NAD-dependent protection of axon degeneration. J Cell Biol 2005, 170:349–355.CrossRef 67. Kaundal RK, Shah KK, Sharma SS: Neuroprotective effects of NU1025, a PARP inhibitor in cerebral ischemia are mediated through reduction in NAD depletion and DNA fragmentation. BIX 1294 datasheet Life Sci 2006, 79:2293–2302.CrossRef 68. Ying W, Wei G, Wang D, Wang Q, Tang X, Shi J, Zhang P, Lu H: Intranasal administration with NAD+ profoundly decreases brain injury in a rat model of transient focal ischemia. Front Biosci 2007, 12:2728–2734.CrossRef 69. Liu D, Pitta M, Mattson MP: Preventing NAD (+) depletion protects neurons against excitotoxicity: bioenergetic effects of mild mitochondrial uncoupling and caloric restriction. Ann N Y Acad Sci 2008, 1147:275–282.CrossRef 70. Wang S, Xing Z, Vosler PS, Yin H, Li W, Zhang F, Signore AP, Stetler RA, Gao Y, Chen J: Cellular NAD replenishment confers marked neuroprotection against ischemic

cell death: role of enhanced DNA repair. Stroke 2008, 39:2587–2595.CrossRef Competing Resveratrol interests All authors declare that they have no competing interests. Authors’ contributions LL, JZ, YY, QW, YC, ZS, MZ, and GG have carried out the molecular genetic studies, participated in the sequence alignment, and drafted the manuscript. LL, JZ, LG, YY, TC, XZ, GX, and GG participated in the design of the study and performed the statistical analysis. JZ and GG conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.”
“Review Introduction Liposomes are small artificial vesicles of spherical shape that can be created from cholesterol and natural non-toxic phospholipids. Due to their size and hydrophobic and hydrophilic character(besides biocompatibility), liposomes are promising systems for drug delivery. Liposome properties differ considerably with lipid composition, surface charge, size, and the method of preparation (Table  1). Furthermore, the choice of bilayer components determines the ‘rigidity’ or ‘fluidity’ and the charge of the bilayer.

jejuni in the chicken gut and as such, bacteria that do not bind to smaller sugars would potentially have a competitive advantage. Conclusions The conclusions drawn from the initial screening of C. jejuni 11168

on our glycan array [3] have in the main been confirmed by the screening of additional strains. Sialic acid and https://www.selleckchem.com/products/ew-7197.html mannose still appear to be the general structures recognised after environmental stress, appearing to be important for initial host pathogen interactions. Galactose and fucose structures still appear to be crucial for the persistence of infection. Little difference is seen between the isolates from clinical or chicken hosts, with the exception of carageenan and branched mannose binding, with both more likely to be recognised by chicken isolates than those isolated from

humans. This study increases the understanding of C. jejuni glycan recognition and provides a model for the study of complex glycan recognition from a PLX-4720 purchase number of other yet to be screened bacterial species. Methods Bacterial strains and growth conditions The strains used in this study can be found in Table 5. Bacteria were grown as previously described [3]. Table 5 Bacterial strains used in this study Strain Invasive Source Human +/−   11168 + D. Newell 351 + RMIT 375 + RMIT 520 + RMIT 81116 + D. Newell 81-176 + J. G. Fox Chicken     331 – RMIT 8 + RMIT 19 + RMIT 108 + RMIT 434 + RMIT 506 + RMIT Glycan arrays Glycan arrays were prepared and selleck screening library performed as previously described by Day et al.[3] with slight modification to the preparation of the slides as outlined by Hartley-Tassell et al.[30] using the glycan library described in Arndt et al.[3, 30, 31]. See Additional file 1: Table S1 for full list and DOK2 structures of glycans. The arrays were scanned

by a ProScan Array scanner at 488/520 nm and the results analysed by ScanArray Express software program. Binding was defined as a value greater than 1 fold increase above mean background relative fluorescence units (RFU). The mean background was calculated from the average background of empty spots on the array plus three standard deviations. Statistical analysis of the data was performed by a Student’s t-test with a confidence level of 99.99% (p ≤ 0.0001). All arrays were performed in triplicate with a total of 12 data points for each glycan tested. Lectin competition adherence assays Adherence and lectin competition assays were performed as previously described [3], however, only using C. jejuni grown at 37°C under micraerobic conditions. E. coli DH5α cells were used as a control for the lectin competition assays to ensure that reduction in adherence was not due to steric hindrance of the lectins on the cell surface inhibiting cell binding to non-glycan targets. Lectins were used at 10 μg per well. All assays were performed in triplicate. Free glycan inhibition assay Adherence assays were performed as previously described [3] under conditions described above.

Through its experience in renewable energy technologies, AuroRE is also offering its services to European companies looking to certify and carry out field inspections on renewable energy projects and carbon emission reduction

projects and programs for their Indian clients (Lamba 2009; Shekhar 2009). THRIVE has generated revenues of around USD 2 million till now. THRIVE is developing a renewable energy center outside Hyderabad for training and demonstration projects in renewable energy. It has plans to start new programs for rural water treatment, rural electrification, rural banks, and rural village outlets. THRIVE also has plans to enter into the solar power generation business in line with the National Solar Mission of

the Government of India. In addition, THRIVE is helping many corporate organizations to implement corporate social responsibility (CSR) programs in relation to LED lighting #VE-822 ic50 randurls[1|1|,|CHEM1|]# (Ramani 2010; THRIVE 2011). NEST is planning to expand its production, warehousing, and marketing and sales capabilities through an investment of around BMN 673 price INR 60 million. It expects revenues of around INR 543 million by 2014–2015 and is targeting an EBIDTA (earnings before interests, taxes, depreciation, and amortization) of around 25 % from the fifth year onwards, i.e., from 2015. Mr. Barki is also planning the manufacturing of solar panels in China to reduce costs (Barki and Barki 2010; Uppal and Mahendra 2009; NEST 2009). D.light Design, on the other hand, is focused on becoming a truly global PAK5 company. D.light Design has grown to over 70 employees in three years and has

offices in the USA, India, Tanzania, China, and Hong Kong. In 2010, D.light Design centralized its product design and international sales in Hong Kong, with plans to move additional corporate functions (D.light 2010, 2011). Geographical upscaling With regard to geographical upscaling, there are unique patterns that are dependent on the chosen business model. SELCO is focusing on expanding geographically in five Indian states neighboring Karnataka, including Maharashtra, Tamil Nadu, Kerala, and Andhra Pradesh. By the end of financial year 2010–2011, it is expected that SELCO would be present in 16 districts of Karnataka, 3 districts of Kerala, 4 districts of Gujarat, and 3 districts in states like Maharashtra and Andhra Pradesh (SELCO 2009, 2010). However, SELCO has found it difficult to expand geographically across different Indian states due to the lack of spillover learning across different states and the lack of financial institutions with whom SELCO can partner with. At the same time, SELCO does not want to use the franchise system to sell its products and services, as the reputation of its brand depends on services and it is more difficult to guarantee the same quality of service from franchises.