Considerable efforts were made to ensure that neither the patient

Considerable efforts were made to ensure that neither the patients nor the dentist performing the tests were aware of which group and sequence the child Dasatinib order was allocated to. Blinding of the chair-side assistant was not possible, as

she was administering the drugs. The patients could probably have been aware of the sedative effect of inhalation of N2O/O2. As this is part of the pharmacological effect of the dug, it could not be disguised, but patients were carefully instructed, not to communicate with the dentist performing the tests. Furthermore, the dentist only entered the operatory, performed the tests and left the operatory again without having any communication with the patients. Thus, bias due to these factors seems to have been reduced as much as practically possible. A suggestion for further studies could be to have asked the participants to guess whether they had received placebo or N2O/O2 NVP-LDE225 in vitro as a check of the blinding. The present study was conducted as a crossover trial with random allocation to two sequences. The strength of this design

is usually considered to be an increase in statistical power, as the patient is serving as his/her own control. Power calculations performed prior to the study based on pilot data from children from the same population and of the same age showed that a minimum of 28 patients in each group was needed to identify a 25% reduction in tooth-pulp pain sensitivity for α = 0.05 and β = 0.80. Power calculations also showed that approximately 200 individuals in each group would be necessary to obtain the same power in a parallel group design. Recruiting children of 12–15 years for at study like the present proved to require considerable effort and time. Furthermore, it required complicated negotiations with authorities to obtain the required

approval for the study. Thus, any reduction in number of subjects needed Sinomenine can save considerable resources. In spite of the fact that N2O/O2 inhalation is commonly seen as a successful method to obtain acceptance of restorative treatment in children and adolescents, the present study has not been able to show any analgesic effect on tooth-pulp pain sensitivity, but did find a 20% reduction in pressure-induced pain of the jaw muscles, Thus, the success of N2O/O2 inhalation in restorative paediatric dental care must also be caused by other factors. First of all, the sedative effect would result in a more relaxed patient, who would react later – and maybe less precisely – on painful treatment. This is supported be the finding that the discomfort of the children from the two experimental tests was not influenced by the inhalation of N2O/O2. Secondly, many of the other unpleasant stimuli, the patient received during restorative treatment, like muscle discomfort from having to keep the mouth open for a long time, etc. may be less disturbing when sedated.

2b Although all investigated bacteria possess a PPDK, only Anaer

2b. Although all investigated bacteria possess a PPDK, only Anaerocellum thermophilum [recently reclassified as Caldicellulosiruptor bescii (Yang et al., 2009)] reveals the same gene arrangement as C. saccharolyticus. GSK1120212 mouse For A. thermophilum, an additional ORF, coding for a hypothetical protein, can be found overlapping both the PPDK and the DeoR ORF. Whether the PPDK gene clusters of C. saccharolyticus and A. thermophilum are transcribed as a single polycistronic mRNA remains to be investigated. In contrast, PPDK

from Thermotoga maritima clusters with the glycolytic enzyme FBA, an acetate kinase and a GntR-type transcription regulator (data not shown). Furthermore, except for Clostridium thermocellum, which lacks a PK, all the investigated organisms revealed the PK gene to be clustered with the gene coding Roxadustat ic50 for the ATP-PFK, suggesting coregulation (Belouski et al., 1998). In Lactococcus lactis, the ATP-PFK and PK operon additionally contains the gene coding for LDH and is known as the las (lactic acid synthesis) operon (Llanos et al., 1993). If PPDK acts in the catabolic direction, C. saccharolyticus has two options for converting PEP to pyruvate

and ATP. It is therefore plausible that some type of regulation might occur. Therefore, the influence of PPi levels on PK activity in C. saccharolyticus was investigated. PPi was found to inhibit PK activity in C. saccharolyticus, with an apparent Ki value of 2.9 ± 0.9 mM PPi (Fig. 4). Consequently, when the PPi levels are high during exponential growth (approximately 4 mM;

Fig. 3), the PK is inhibited by ∼60%, again suggesting a catabolic role for PPDK in this growth phase. Consistently, in Trypanosoma cruzi, where PPDK is also working in the direction of ATP generation, PPi is also a strong inhibitor of PK (Acosta et al., 2004). Furthermore, PPDK has been shown to be used in the direction of ATP synthesis in some other organisms (Tjaden et al., 2006; Feng et al., 2008). The role of PPi as an allosteric effector has recently also been described for the LDH of C. saccharolyticus (Willquist & van Niel, 2010). PPi acts as an inhibitor of the LDH, while ATP stimulates the enzyme. The estimated kinetics of the LDH explains the Baricitinib switch from a metabolism producing mainly acetate to a metabolism producing less acetate and more lactate. The hydrolysis of PPi is generally regarded as an indispensable reaction of a cell’s metabolism. PPi is a byproduct of various energy-requiring biosynthetic reactions, for example DNA and RNA synthesis and during the formation of precursors for protein and polysaccharide synthesis (Heinonen, 2001). These reactions are often close to equilibrium and only the effective removal of PPi drives these reactions forward. Therefore, the coupling of these reactions to PPi hydrolysis is crucial to maintain growth (Chen et al., 1990). It is unknown what levels of PPi still allow the cellular metabolism to proceed, but apparently, C.

3E–E6) suggests that these cells first migrate caudally in the

3E–E6) suggests that these cells first migrate caudally in the

lateral subpallium before turning, and migrating in the lateral-to-medial direction within the EA. In sum, our analysis revealed that scgn+ cells cytoarchitecturally resembling migrating neurons formed a continuous stream along the palliosubpallial boundary before reaching their final destinations in the OB or EA (Fig. 4). Next, we analyzed the distribution of scgn+ neurons find more in neonatal mouse brain. We observed that the migration of scgn+ cells concluded by birth and scgn+ neurons inhabited, in an anterior-to-posterior direction, the spatially interrelated nuclei of the BST, interstitial nucleus of the posterior limb of the anterior commissure (IPAC), ventral pallidum (VP), dorsal substantia innominata (SI), and the central and medial amygdaloid nuclei (CA and MA; Fig. 5A–A7). Morphometric analysis revealed that scgn identifies divergent neuron

subpopulations with different somatic diameters in the VP and EA (Fig. 5B). By using genetically tagged reporter mice we demonstrated that scgn+ neurons either adopted a GABA phenotype along the longitudinal axis of the EA (Fig. 5D and D1), similar to scgn+/GABA+ neurons in Selleck Trametinib the embryonic OB (Fig. 5C and C1), or co-express ChAT when found in small-diameter cholinergic neurons of the dorsal SI (Fig. 5E and E1). Collectively, our data suggest that by E18 scgn+ neurons can acquire a distribution pattern resembling that in the adult brain, and differentiate into neurochemically distinct subtypes of EA

neurons. Systematic analysis along the longitudinal axis of the fetal primate brain revealed the first contingent of scgn+ neurons in the granular and glomerular layers of the OB (Fig. 6A). However, unlike in the adult primate brain (Mulder et al., 2009b), neuroblasts migrating in the prenatal rostral migratory stream (Pencea & Luskin, 2003) did not harbour scgn expression (Fig. 6A). Pallial areas were devoid of discernible Montelukast Sodium scgn immunoreactivity. In the basal forebrain, scgn+ neurons were seen in the horizontal diagonal band, nucleus accumbens, medial septum, VP, GP and SI (Fig. 6A1–A7). In contrast to scgn distribution in the neonatal rodent brain, scgn+ cells were only infrequently found in either the CA or MA. In the hypothalamus, substantial scgn+ neuron populations occupied the paraventricular and periverticular nuclei and the supraoptic nucleus (Fig. 6A5–A7). A morphological dichotomy of scgn+ neurons was evident in the basal telencephalon (Fig. 6B): small-to-medium-sized scgn+ neurons populated the horizontal diagonal band, SI and CA. In contrast, large-diameter scgn+ neurons were found in the IPAC and GP.

tumefaciens GV3101∷pMP90 to obtain the strain GV3101∷pMP90(pPZP-e

tumefaciens GV3101∷pMP90 to obtain the strain GV3101∷pMP90(pPZP-eGFP). The vector pRK415 GSK J4 clinical trial (Keen et al., 1988) or the plasmid pRKLACC (Shah et al., 1998), which is pRK415 containing the acdS gene from Pseudomonas putida UW4 under the control of a lac promoter, was electroporated into A. tumefaciens GV3101∷pMP90(pPZP-eGFP) to obtain strain YH-1, which is GV3101∷pMP90(pPZP-eGFP)(pRK415), and strain YH-2, which is GV3101∷pMP90(pPZP-eGFP)(pRKLACC). Agrobacterium strains were grown in Luria–Bertani (LB) (Miller, 1976) or M9 medium (Atlas, 1993) (for ACC deaminase

activity assay) at 28 °C. When required, antibiotics were added at the following concentrations: rifampicin, 50 μg mL−1; gentamicin, 50 μg mL−1; spectinomycin, 50 μg mL−1; streptomycin, 20 μg mL−1; and tetracycline, 2 μg mL−1. An ACC deaminase activity assay was performed as described by Hao et al. (2007). The infection and regeneration protocols were modified from Cardoza & Stewart (2003). The media used are listed in Table 1. Seeds of B. napus cv. Westar, B. napus cv. Hyola 401 and B. napus cv. 4414 RR were surface sterilized by soaking in 70% ethanol for 1 min, followed by 20% commercial bleach for 20 min, and were then

rinsed four times with sterilized distilled water and planted at a density of 10–12 seeds per Petri dish (100 × 25 mm) (Fisher Scientific, Ottawa, ON) on seed germination medium. Seeds were germinated at 22–25 °C in the dark for about 1 week. The seedling drug discovery hypocotyls were cut into about 1-cm pieces and preconditioned for 3 days on a cocultivation medium. Agrobacterium tumefaciens strains YH-1 and YH-2 were grown in 50 mL LB medium until the culture reached OD600 nm≈1. The cells were pelleted, resuspended in the infection medium and normalized to OD600 nm=1 to obtain a 1 × dilution. Serial dilutions were then performed using the infection medium to obtain 10−1× and 10−2× dilutions. The preconditioned explants were infected by soaking in A. tumefaciens culture suspensions for 30 min at room temperature with gentle shaking. The infected hypocotyls were first

cocultured on a cocultivation medium for 48 h, then transferred to a callus induction medium for 2 weeks, Palmatine then to an organogenesis medium with (OA) or without AgNO3 (OB) for another 2 weeks, and then to a shoot induction medium for 3–6 weeks until shoots appeared. The induced shoots were transferred to a shoot elongation medium for 2 weeks and then to a rooting medium for another 2 weeks, and finally, the transgenic plants were transferred to soil and grown in a greenhouse. Plant tissue cultures were maintained in a growth chamber at 25 °C with 16 h of light and 8 h of dark, with a light intensity of 40 μmol m−2 s−1 from cool-white fluorescent lamps. The stable transformation frequency was calculated using the following formula: transformation frequency=the number of transgenic plants obtained/the number of explants used for transformation. After infection with various dilutions of A.

The result suggests that a small fraction of the pLS32neo molecul

The result suggests that a small fraction of the pLS32neo molecules, which had escaped the BsuM restriction, settled in the R+ M+ cell together with pHV33 in the same way as observed for see more the transfer in the homologous pairs. When the donor was proficient in the BsuM

function and the recipient was not, the fractions of the colonies showing Spr Nmr Cmr were 8% and 10% among those showing Spr Nmr and Spr Cmr, respectively (line 4 in the last two columns). The percentages were 1/9 to 1/7 of those observed for the homologous pairs. The above-mentioned results suggested the usefulness of the restriction-deficient B. subtilis protoplast as a host for successful transfer of genetic materials from other bacterial species. This notion prompted us to test the R− M− RM125 strain for interspecific cell fusion with two strains of bacilli, one a thermophile, B. stearothermophilus, and the other a mesophile, B. circulans. The protoplasts of B. stearothermophilus CU21 Doxorubicin and B. circulans BM carrying pTHT151 (Tcr) and pHB201ds15dlt (Cmr), respectively, were fused with those of B. subtilis RM125 recA::Emr. It was shown that the plasmids were successfully transferred from the donor strains to B. subtilis

RM125 (Table 3), although the transfer efficiencies were 1/7 to 1/5 as compared with the fusion between the B. subtilis Inositol monophosphatase 1 R+ M+ (donor) and R− M− (recipient) (Table 2, line 4). It has been reported that Type I restriction enzymes are located at different cytoplasmic membrane sites of the Escherichia coli cell (Holubova et al., 2004). The current study demonstrates that the BsuM restriction enzyme is present at least in part in the cytoplasm, because pLS32neo with eight BsuM restriction sites was restricted

upon cell fusion, which involves the contact of the cytoplasms from the donor and the recipient cells. It was shown that pLS32neo was severely restricted upon transfer from the R− M− to R+ M+ cells, whereas its transfer from the R+ M+ to R− M− cells was successful, although the efficiency was lower (7.8–8.8%) than that for the transfer between the R− M− donor and recipient pair (see ‘Results’). The reduced but significant transfer efficiency from the R+ M+ to R− M− cells indicates that the chromosomal DNA in the recipient R− M− cell survived the attack of BsuM restriction from the cytoplasm of the donor R+ M+ cell. How can these phenomena be explained? There may be two possible explanations. One is that the fusion of multiple protoplasts of the recipient R− M− cells with a donor R+ M+ protoplast carrying pLS32neo diluted the BsuM enzyme level in the fusant, resulting in successful transfer of the plasmid. This explanation, however, is unlikely because a similar situation, i.e.

Anti-HBs antibody GMCs at year 4 were 423, 236, and 137 mIU/mL

Anti-HBs antibody GMCs at year 4 were 42.3, 23.6, and 13.7 mIU/mL in the three groups, respectively. One month after the additional dose of hepatitis A-containing vaccine, ≥99.4% of subjects in all

the groups were seropositive RG7204 in vivo for anti-HAV antibodies. Anti-HAV response rates were 98.2% in the HAB group, 97.6% in the ENG + HAV group, and 99.4% in the HBVX + VAQ group. One month after the additional dose of hepatitis B-containing vaccine, 95.2% of subjects in the HAB group had antibody concentrations ≥10 mIU/mL compared with 90.5 and 85.3% in the ENG + HAV and HBVX + VAQ groups (p = 0.1367 and p = 0.0026, respectively) (Figure 1B). Corresponding anti-HBs GMCs were 7233.7, 1242.5, and 1075.1 mIU/mL. Overall anti-HBs response rates were 93.4% in the HAB group, 88.1% in the ENG + HAV group, and 83.4% in the HBVX + VAQ group (p = 0.1305 and p = 0.0054, respectively). In subjects with anti-HBs antibody selleck inhibitor concentration <3.3 mIU/mL prior to administration of the additional vaccine dose, 82.1, 82.0, and 72.6% achieved an anti-HBs concentration ≥10 mIU/mL post-vaccination. In the three groups, virtually all seronegative subjects who failed to respond to the additional dose and reach the cut-off of 10 mIU/mL were already nonresponders, or very low

responders, to primary vaccination. Exploratory subgroup analyses showed hepatitis A and B seropositivity rates at year 4 to be slightly lower in subjects aged ≥61 years, with a BMI ≥30 kg/m2, receiving concomitant medication or with a current concomitant medical condition. No consistent effects of any of these factors on response to the additional vaccine dose(s) were observed (data not shown). We assessed persistence of immune response to a combined hepatitis A/B vaccine in adults aged >40 years. The study population can be considered representative of the general

population in this age group, with a high proportion of subjects being overweight, having underlying Phospholipase D1 medical conditions, or receiving concomitant medication. The differences in immune response between the combined hepatitis A/B vaccine and monovalent vaccines previously observed after primary vaccination6 were found to be maintained over time. As described in another follow-up study of this combined hepatitis A/B vaccine in this population,7 anti-HAV seropositivity rates remained high in all groups over the 4 years of follow-up. At year 4, anti-HBs rate ≥10 mIU/mL and anti-HBs GMCs were highest in subjects who received the combined vaccine, although these were lower than have been reported in younger adults 10 years after administration of this vaccine.8 The lower anti-HBs antibody response rates observed in the HBVX + VAQ group at all time-points may be due to the reduced HBsAg content and adjuvant composition of the hepatitis B vaccine in this group.

The viability of the ΔcymR mutant and the parental strain was fur

The viability of the ΔcymR mutant and the parental strain was further tested 10 min after the addition of 1 mM H2O2. A three- and sevenfold reduction in survival was observed for the ΔcymR mutant as compared with the wild-type strain grown in minimal medium in the presence of methionine or in LB medium, respectively. These results showed that CymR inactivation led to an increased sensitivity to peroxides

and superoxides. The intracellular cysteine level is maintained within a narrow range to address both the cysteine supply for protein synthesis and Buparlisib clinical trial the production of other essential molecules, and the necessity to maintain cysteine levels below the toxicity threshold. In B. subtilis, the CymR regulator plays an essential role in maintaining intracellular cysteine levels. In a ΔcymR mutant, the derepression of genes involved in cysteine uptake and biosynthesis (Even et al., 2006) leads to an intracellular accumulation of cystine and cysteine and to an increase of H2S production. In this mutant, the sixfold increase in H2S production is probably due to cysteine accumulation and its degradation by cysteine desulfhydrases. Four different cysteine desulfhydrases have been detected in vitro in B. subtilis: MccB, MetC, PatB and CysK (Auger et al., 2005). In the zymogram, we mainly observed an increased

MccB activity in the ΔcymR mutant as compared with the wild-type strain, in agreement with the

derepression of mccB transcription in this mutant (Even et al., 2006). However, TGF-beta activation the mutation in one of the genes encoding cysteine desulfhydrases, either patB or mccB or cysK, was unable to abolish the H2S production in a ΔcymR background (data not shown). This suggests that several enzymes are required for H2S production in vivo, including the possible involvement of a new yet uncharacterized enzyme. The ΔcymR mutant poorly C1GALT1 grows in a minimal medium containing cystine at least partially due to the accumulation of thiol-compounds (cysteine, homocysteine, H2S). In Escherichia coli, cysteine toxicity is mainly related to the inhibition of branched-chain amino-acid synthesis. A previous work indicated that the threonine deaminase, homoserine dehydrogenase and/or acetohydroxyacid synthase are probable target enzymes for cysteine toxicity (Kari et al., 1971; Harris, 1981). Interestingly, we observe a depletion of leucine and valine in the ΔcymR mutant grown with cystine. The addition of these two amino acids enhanced the growth of the ΔcymR mutant, but did not fully restore its growth capacity. The addition of casein hydrolysate did not further improve the growth (data not shown), and even in LB medium, the growth yield of the ΔcymR mutant decreased as compared with the wild-type strain. This suggests that additional toxic effects are mediated by cysteine or derived compounds.

[8] There is no such position in the USA Furthermore, technician

[8] There is no such position in the USA. Furthermore, technicians in the UK can work in ‘ward-based management roles’ in the hospital setting.[6] This involves reviewing drug charts and prescriptions for drug therapy problems, which are then referred to a pharmacist for modification if necessary.[6] In addition to this role there are numerous other management positions which may be held by technicians in the UK. These include dispensary team leader, store and distribution senior technician, and pharmacy clinical trials coordinator, to name a few.[6] In the UK, pharmacy technicians can also work in a clinical

pharmacy technician position. This role involves liaising with other healthcare professionals and having close contact with patients. Clinical pharmacy technicians are given Thiazovivin ic50 responsibilities of discussing and checking patient medications, as well as advising them on the safe and most efficient use of medications.[9] In sum, although the job title Pharmacy technician is used both in the UK and USA, the duties and responsibilities seem to vary significantly. In general, roles for pharmacy technician in the UK are more sophisticated and advanced than in the USA.[6] Rouse et al.

define a pharmacy technician as ‘. . .  find more an individual working in a pharmacy [setting] who, under the supervision of a licensed pharmacist, assists in pharmacy activities that do not require the professional judgment of a pharmacist’.[10] While this is a representative definition, it can vary by setting; a consensus definition remains elusive.[11] Pharmacy technicians work in a multitude of settings, with the majority (75%) employed by community pharmacies[12] where they are involved with nearly 96% of prescriptions dispensed

there.[2] Approximately 16% of technicians work in hospitals/health this website systems with the remaining number employed by long-term care facilities, home healthcare agencies, mail-order pharmacies, managed care organizations and health insurance companies.[13,14] Nine out of ten community pharmacies employ pharmacy technicians, while this number approaches 100% in hospital pharmacies.[15,16] According to the Bureau of Labor Statistics there are 326 300 pharmacy technicians in the USA, whereas the National Association of Boards of Pharmacy (NABP) suggests there may be 414 000 in the USA and Puerto Rico.[17] Professional pharmacy organizations such as the American Society of Health-System Pharmacists (ASHP) and American Pharmacists Association (APhA) are among the trailblazers advocating the use of and standardized training for pharmacy technicians. One goal has been to differentiate between the tasks of professional and non-professional staff in both hospital and community pharmacy settings.

Acinetobacter baumannii clones resistant to phage AP22 were forme

Acinetobacter baumannii clones resistant to phage AP22 were formed at the rate of 10−6 per a cell. A total of 50 phage-resistant clones of

A. baumannii 1053 were analyzed to determine whether they are phage-resistant mutants or lysogens with inserted prophage. To reveal possible spontaneous induction, bacterial suspensions of each clone treated with chloroform were spotted on bacterial lawn of sensitive strain. Besides, the resistant clones were grown in the presence of different concentrations of mitomycin C to show possible presence of the phage in concentrated preparation by EM procedure. In both cases, there was no presence of the phage in the samples. A possibility of the prophage presence in genomic DNA of resistant Staurosporine datasheet click here clones was estimated by PCR with two pairs of primers specific to the phage DNA. It was shown the absence

of prophage DNA in genomic DNA of resistant clones (Fig. 5). Lytic activity and host specificity of the phage were tested against 130 identified A. baumannii genotype-varying MDR strains. These strains were isolated from patients of burn units, units of selective and emergency surgery, therapeutics units, intensive care units, and urology units in 2005–2010. Most of them were resistant to diverse groups of antibiotics, including aminoglycosides, fluoroquinolones, third- and forth-generation cephalosporins, and also cefoperazone sulbactam and carbapenems. All strains were divided into 10 groups by RAPD analysis. RAPD groups A1 and B1 predominated with 48% and

35% of the investigated strains, respectively, and were spread in clinics of a variety of Russian cities. Unlike some other known A. baumannii phages, bacteriophage AP22 was found to have a broad range of lytic activity against A. baumannii multidrug-resistant clinically relevant strains. The phage was shown to specifically infect and lyse 68% (89 of 130) of A. baumannii strains by forming clear zones. Of (-)-p-Bromotetramisole Oxalate particular interest is that the phage lysed 83% (88 of 106) of A. baumannii strains from those two RAPD groups that were dominating in some Russian hospitals between 2005 and 2010 (Table 1). Wound, tissue sampling, sputum, bronchopulmonary lavage, pleural fluid, urine, bile, blood, and hospital environmental rinses Chelyabinsk, Moscow, St. Petersburg Wound, tissue sampling, sputum, bronchopulmonary lavage, pleural fluid, urine, bile, blood, rinses of drainage and intravenous catheters, and hospital environmental rinses Chelyabinsk, N-Novgorod, Moscow, St. Petersburg Chelyabinsk, N-Novgorod, Moscow, St. Petersburg Wound, sputum, and rinses of intravenous catheters Chelyabinsk, N-Novgorod, St. Petersburg The phage was also tested against some other representatives of the genus Acinetobacter (A. lwoffii, A. anitratus, and A. calcoaceticus), as well as several other gram-negative microorganisms such as P. aeruginosa, E. coli, Y. pseudotuberculosis, Y. enterocolitica, K.

YS from the Kearney

Y.S. from the Kearney Galunisertib in vivo Foundation of Soil Science and the faculties of UCR and UofA. The authors gratefully acknowledge M.G. Klotz, B.D. Lanoil, and anonymous reviewers for critical comments on this and previous versions of the manuscript. Fig. S1. Growth curves of AOB cultivated in HEPES- (a) and phosphate- (b) buffered medium; Nitrosomonas

europaea (squares), Nitrosomonas eutropha (circles), and Nitrosospira multiformis (triangles). Table S1. Genes and PCR primers used in this study. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Three indigenous isolates of Bacillus sphaericus (ISPC-5, ISPC-6 and ISPC-8), along with standard 2362 and 1593 strains, were evaluated for spore viability Selleck Quizartinib and mosquitocidal activity. Among these, ISPC-8 was the most viable and virulent isolate, exhibiting a significantly higher total viability count (TVC) and lower

LC50 values. The TVC of the standard strains ranged from 4.0 to 9.2 × 108 spores mL−1, whereas it was 1.3 × 109 spores mL−1 for ISPC-8. The LC50 values of ISPC-8, 2362 and 1593 against Culex quinquefasciatus were 0.68 × 103, 1.22 × 103 and 1.85 × 103 spores mL−1, respectively. The ISPC-8 was further assessed for host spectrum and found to be more active against C. quinquefasciatus, followed by Culex tritaeniorhynchus, Aedes albopictus and Aedes aegypti. The ISPC-8 strain was thus found to be a promising isolate for developing biopesticides. Among the indigenous strains, only ISPC-8 was found to have binary toxin genes (binA and binB). Comparative sequence analysis revealed that the BinA (41.9 kDa) protein of ISPC-8 differs by one amino acid (R197M), whereas BinB (51.4 kDa) differs by two amino acids (H99P, P174S) as compared with 1593 and 2362 strains. The purified binary proteins of ISPC-8 showed an LC50 value of 6.32 ng mL−1 against C. quinquefasciatus larvae

after 48 h. The adverse environmental effects associated with chemical insecticides have led to the search for alternative methods for controlling different disease-transmitting mosquito species. The aminophylline use of entomopathogenic microorganisms appears to be one of the promising alternatives, and microorganisms such as Bacillus sphaericus and Bacillus thuringiensis ssp. israelensis have been quite effective against different mosquito species (Federici et al., 2007). These two bacteria differ in the nature of their toxins and host range. In general, B. sphaericus is more active against Culex and Anopheles sp., whereas B. thuringiensis ssp. israelensis is more active against Aedes and Culex sp. (Charles et al., 1996). Bacillus sphaericus has an additional attribute as it persists in polluted aquatic environments, whereas in this environment, the toxicity of B. thuringiensis ssp. israelensis is lost rapidly (Silapanuntakul et al.