The study was approved by the University of East Anglia Ethics Committee. An introductory e-mail was sent out to 10,000 e-mail addresses held by the Centre for Pharmacy Postgraduate Education

containing a link to an online survey with a follow up e-mail after two weeks. It was estimated that 1/3rd of e-mail addresses may be no longer active and that only 50 % of the remaining e-mail addresses were for practicing community pharmacists. Participants were asked to enter how many consultations (one to one discussions in the consultation room) they had held with patients in their last standard working week. STATA® 12 SE was ERK inhibitor clinical trial used to conduct a backward stepwise elimination linear regression model for the number of consultations as the dependent variable. A total of 700 responses (42% of predicted potential LGK 974 respondents) with 595 responses eligible for inclusion.

Descriptive results have been reported previously2. The median (quartiles) for the number consultations performed in a standard week was 5 (3, 10), these include Medicine Use Reviews, New Medicine Service and additional enhanced services such as emergency contraception. The statistically significant predictors of number of consultations in the final model were: working in a multiple pharmacy, having received consultation skills training during preregistration, male gender,

requesting further consultation skills training, and greater confidence in consultation skills. Confidence in consultations skills had the highest positive relationship with number of consultations. Participants had to rate their how confident they were in their consultation skills on a scale where 1 was not confident and 5 was fully confident. A value of 3 on the confidence scale was modelled Methane monooxygenase as having an increase of 34% in the number of consultations compared to the reference group of confidence 1 or 2 (p = 0.025); a value of 4 an increase of 56% (p < 0.001) and a confidence rating of 5 an 81% increase on the reference group (p < 0.001). The model explained 27.2% of the variance in the number of consultations. This exploratory analysis suggests that the more confident a participant is in their consultation skills, the more consultations they conduct. Previous research suggests that training is important in increasing confidence3. While there are many changes in pharmacy education to include consultation skills training during undergraduate and pre-registration year, there are still a large number of registered pharmacists for whom further training in consultation skills could help increase the delivery of more patient facing services.

melbae and C. columbae samples, respectively. We thank

the Slovak Academy of Science and IAEA for tsetse pupae. We acknowledge the funding support of NASA NNX07AL53A, NIH R03AI081701 and NSF-REU DBI-0849917. Table S1. Primers, annealing temperatures (Ta), and resulting amplicon sizes. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials PD-0332991 nmr supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Genetic transformation is an indispensable tool for basic fungal research, as well as a useful technique for directed improvement of industrial strains. Here we describe a simple and reproducible transformation system for the filamentous fungus Hypocrea jecorina. The system is based on hxk1 (encoding hexokinase) as selectable marker, a hexokinase-negative strain and d-mannitol, which is used as selective carbon source and osmotic stabilizer. Following transformation with the hxk1 gene, the obtained transformants were able to grow on d-mannitol as sole carbon source. Transformation efficiency achieved using d-mannitol as carbon source and osmotic stabilizer was roughly five

times higher than that using d-sorbitol. The utility of this system was further demonstrated by transformation of H. jecorina with the egfp (encoding the enhanced green fluorescent protein) gene. Fluorescence microscopy revealed EGFP fluorescence Ergoloid in positive transformants. Our results demonstrated the feasibility of exploiting a carbon source metabolic selleck chemicals llc pathway for the development of promising fungal transformation systems, which provides a new molecular toolbox for genetic modifications of the cell factory H. jecorina. Hypocrea jecorina (anamorph Trichoderma reesei) is one of the workhorse organisms for production of a wide spectrum of polysaccharide-hydrolyzing enzymes, including

cellulases and xylanases, which are applied today in the food, pharmaceutical, textile and pulp industries. Hypocrea jecorina is also recognized as a model cellulolytic microorganism and research efforts today are focused on understanding and improving cellulase efficiency and productivity (Hartl et al., 2007; Seiboth et al., 2007; Fekete et al., 2008; Martinez et al., 2008; Stricker et al., 2008). Moreover, due to its enormous secretion potential and its generally regarded as safe status, H. jecorina is considered an attractive cell factory for large-scale production of homologous and heterologous proteins (Nyyssonen et al., 1993; Nevalainen et al., 2005). The genetic transformation of filamentous fungi is a crucial prerequisite for manipulations at the molecular level. Techniques suitable for H. jecorina transformation, such as protoplast transformation (Gruber et al., 1990), biolistic transformation (Te’o et al., 2002) and Agrobacterium tumefaciens-mediated transformation (Zhong et al.

Most of these requests were ordered from the hospital’s emergency department for suspected insufficient serum concentrations. Antiepileptic drug monotherapy is still the most frequently employed therapeutic strategy in adult patients with epilepsy in keeping

with the standard therapeutic guidelines. Sodium valproate is commonly used for different types of seizures reflecting its wide spectrum of anticonvulsant potential. Newer AED utilizations are becoming increasingly popular in our subjects particularly as add-on with other standard AEDs. “
“Objectives  To determine statin usage pattern and evaluate whether new generation statins are actually needed by the patients receiving them. Methods  Cisplatin price This retrospective cohort included patients receiving first-time statins at a tertiary care hospital in Thailand. Using electronic medical records from 2005, its indication was determined based on history of coronary heart disease (CHD) and CHD-risk equivalents. The lipid profiles tested within 30 days prior to the first date of statins prescription were analysed. Each patient was assessed as

selleck kinase inhibitor to whether statin was needed based on low-density lipoprotein cholesterol (LDL-C) reduction capacity and lipid goals. Results  A total of 2479 first-time statin users was included. Ninety percent of the users received simvastatin, while 8% and 2% received atorvastatin and pravastatin respectively. More than half (58.0%) used statins for primary prevention, although all usage of atorvastatin was considered not needed. Considering the use of statin for secondary prevention to achieve the LDL-C goal of <130 mg/dl (3.37 mmol/l), more than 80% of atorvastatin users could be switched to simvastatin. Only 8% of simvastatin Megestrol Acetate usage would not be able to achieve this target. When

the LDL-C goal was <70 mg/dl (1.81 mmol/l), 40.2% simvastatin users was considered appropriate, while 58.6% needed atorvastatin to be prescribed. Conclusion  A substantial proportion of patients did not need statins therapy, particularly for primary prevention. In addition, atorvastatin use is mostly not needed except in patients requiring statins for secondary prevention to achieve the LDL-C goal of <70 mg/dl (1.81 mmol/l). The findings should prompt hospital policy makers to develop measures to ensure the proper use of statins in their clinical settings. "
“Objective  Most epilepsies are managed with anti-epileptic drugs (AEDs), but medication non-adherence has been frequently reported. Satisfying patient information needs has demonstrated improved adherence. Multi-professional working has been encouraged to provide cost-effective health services by using the most appropriate healthcare professional. Research has demonstrated that pharmacist-led consultations are acceptable to patients with other medical conditions and therefore may be appropriate for patients with epilepsy.

cerevisiae and Aspergillus fumigatus) revealed the presence of two distinct regions. The one

located at the 5′- region showed high homology with the Spe genes, whereas the one present at the 3′-region was homologous to the Sdh genes; both were linked through a region of approximately 60 nucleotides without Selleck Selumetinib homology (not shown). As expected, the alignment of amino acid sequences encoded by these genes showed the same pattern of homology, demonstrating the high preservation of the gene in the Basidiomycota (not shown). With these data we designed degenerate primers to be used for PCR amplification of the chimeric genes. The forward primer was selected at the 3′-end of the region with homology to Spe, and the reverse primer was designed from the homologous region

at the 5′-end of the Sdh, in such a way that the amplification fragment covered the nonhomologous region that separates both coding regions (see Fig. 1a). Using the PCR conditions described above and selleck chemical the designed degenerate primers, it was possible to amplify DNA fragments of the predicted size from genomic DNA of all the Basidiomycota species tested (see Materials and methods), whose genomes have been sequenced or not, that represented the three subphyla from Basidiomycota. The size of the fragments (around 1300 bp) coincided with the expected values. On the other hand, and as expected, no such amplification occurred when DNA from Ascomycota or Zygomycota species was used as template (Fig. 1b). The PCR products corresponding to the Basidiomycota species analyzed in this work were sequenced. Alignment of the encoded sequences revealed their high conservation (Fig. 2). Additionally, the encoded sequences

of the amplified fragments from Basidiomycota species whose genomes had been previously sequenced were compared with those existing in their corresponding data banks. The results obtained confirmed the fidelity of the PCR amplification Enzalutamide clinical trial (Table 1). The differences observed can be explained by the fact that different isolates were used in these studies. The sequences of the fragments were deposited in GenBank, with the following accession numbers: Ustilago cynodontis, FN646089; Tilletia foetida, FN646090; Bjerkandera adusta, FN646091; Rhizoctonia solani, FN822770; Schizophyllum commune, FN822771; Ustilago hordei, FN822772; Ustilago maydis, FN822773; Coprinus cinerea, FN822774; Pleurotus ostreatus, FN822775; Ganoderma lucidum, FN822776; Agaricus bisporus, FN827330; and Ganoderma sp., FN827329. The sequences of the regions corresponding to the fragments amplified by PCR from the Spe-Sdh genes obtained in this study, and those reported in the databases, were used for the construction of a phylogenetic tree. The results obtained showed the phylogenetic relationship (Fig.

The spectral width in the carbon dimension was 170 p.p.m. and 180 p.p.m., respectively. All spectra were processed and analyzed using Bruker’s topspin

(v3.0) software. Usually, zero-filling was applied to double the number of real points in each dimension. Chemical shifts were referenced to the HDO resonance at 4.7 p.p.m. Chemical shift assignments for 13C were determined indirectly from HSQC and HMBC spectra. Pseudomonas sp. strain Chol1 was subjected to random transposon mutagenesis Y-27632 by insertion of the transposon mini-Tn5 Km1 and screened for transposon mutants showing altered growth with cholate as described previously (Birkenmaier et al., 2007). One mutant, strain G12, was analyzed further. Strain G12 could not grow with cholate as the sole substrate, but it could grow with succinate in the presence of cholate. HPLC analysis of supernatants from these cultures revealed that strain G12 did not transform cholate at all. We then checked this website whether strain G12 could grow with intermediates of cholate degradation. With supernatants containing DHADD (VIII), strain G12 could grow after a long lag phase. Notably, cells of strain G12 induced for growth with DHADD were also induced for cholate transformation during growth with succinate in

the presence of cholate. HPLC analysis revealed that cholate was transformed into several compounds with an absorption maximum at 244 nm, which is indicative of steroids with a 3-keto-1,4-diene structure of the A-ring (Philipp et al., 2006). In the next step, we identified the gene in strain G12, in which the mini-Tn5 Km1 had been inserted. The transposon Cyclooxygenase (COX) was inserted into an

ORF of 1212 bp at bp 333. The predicted protein had 403 amino acids and showed high identity to nonspecific lipid transfer proteins from various bacteria. Among these were two bacteria, for which growth with cholate had been demonstrated, namely Pseudoalteromonas haloplanktis strain TAC125 (Birkenmaier et al., 2007) and Comamonas testosteroni strain KF-1 (Rösch et al., 2008). The nonspecific lipid transfer proteins from strains TAC125 and KF-1 showed 80% and 68% identity, respectively, to the gene product from strain Chol1 (Fig. 2). This gene was named skt (for steroid β-ketothiolase) for reasons that will be described below. To investigate the function of skt for cholate degradation further, we decided to construct a defined mutant of this gene by subjecting strain Chol1 to insertional mutagenesis with the suicide vector pKnockoutG. The resulting strain Chol1-KO[skt] could not grow with cholate; growth with cholate was restored when an intact copy of skt was provided in trans on the vector pBBR1MCS-5 (Fig. 3a). This complementation clearly showed that the phenotype of this mutant was caused by the inactivation of skt. Strain Chol1-KO[skt] could grow with succinate in the presence of cholate (Fig. 3b).

The spectral width in the carbon dimension was 170 p.p.m. and 180 p.p.m., respectively. All spectra were processed and analyzed using Bruker’s topspin

(v3.0) software. Usually, zero-filling was applied to double the number of real points in each dimension. Chemical shifts were referenced to the HDO resonance at 4.7 p.p.m. Chemical shift assignments for 13C were determined indirectly from HSQC and HMBC spectra. Pseudomonas sp. strain Chol1 was subjected to random transposon mutagenesis p38 MAPK inhibitors clinical trials by insertion of the transposon mini-Tn5 Km1 and screened for transposon mutants showing altered growth with cholate as described previously (Birkenmaier et al., 2007). One mutant, strain G12, was analyzed further. Strain G12 could not grow with cholate as the sole substrate, but it could grow with succinate in the presence of cholate. HPLC analysis of supernatants from these cultures revealed that strain G12 did not transform cholate at all. We then checked selleck products whether strain G12 could grow with intermediates of cholate degradation. With supernatants containing DHADD (VIII), strain G12 could grow after a long lag phase. Notably, cells of strain G12 induced for growth with DHADD were also induced for cholate transformation during growth with succinate in

the presence of cholate. HPLC analysis revealed that cholate was transformed into several compounds with an absorption maximum at 244 nm, which is indicative of steroids with a 3-keto-1,4-diene structure of the A-ring (Philipp et al., 2006). In the next step, we identified the gene in strain G12, in which the mini-Tn5 Km1 had been inserted. The transposon Paclitaxel order was inserted into an

ORF of 1212 bp at bp 333. The predicted protein had 403 amino acids and showed high identity to nonspecific lipid transfer proteins from various bacteria. Among these were two bacteria, for which growth with cholate had been demonstrated, namely Pseudoalteromonas haloplanktis strain TAC125 (Birkenmaier et al., 2007) and Comamonas testosteroni strain KF-1 (Rösch et al., 2008). The nonspecific lipid transfer proteins from strains TAC125 and KF-1 showed 80% and 68% identity, respectively, to the gene product from strain Chol1 (Fig. 2). This gene was named skt (for steroid β-ketothiolase) for reasons that will be described below. To investigate the function of skt for cholate degradation further, we decided to construct a defined mutant of this gene by subjecting strain Chol1 to insertional mutagenesis with the suicide vector pKnockoutG. The resulting strain Chol1-KO[skt] could not grow with cholate; growth with cholate was restored when an intact copy of skt was provided in trans on the vector pBBR1MCS-5 (Fig. 3a). This complementation clearly showed that the phenotype of this mutant was caused by the inactivation of skt. Strain Chol1-KO[skt] could grow with succinate in the presence of cholate (Fig. 3b).

6%) with splenomegaly observed in 96 patients (33%) and jaundice in 17 (5.8%). The most common laboratory abnormalities observed in our case series were thrombocytopenia (239, 82%), elevated serum lactate dehydrogenase levels (276, 95%), elevations of liver transaminases (96, 33%), and anemia (89, 30%); only five patients (1.7%) had a hemoglobin level below 80 g/L. Plasmodium vivax-infected patients had lower mean platelet counts than P falciparum-infected subjects (86 × 109/L vs 97.9 × 109/L; p = 0.02). Quantification of parasites by direct microscopy was available on admission for 145 patients (49.8%) of whom 117 with P falciparum malaria (50%) and 28 (49.1%) with

P vivax/P ovale; in more detail, for the former, parasite counts ranged Navitoclax from 68/µL to 1,652,000/µL (median value 60,600/µL) with 26 patients showing a parasite count of more than 5% (median 338,000/µL, range 253,460–1,652,000/µL). For the latter, parasitemia ranged from 210/µL to 57,600/µL (median 1,340/µL). Of the 233 patients with P falciparum malaria, 35 (15%) fulfilled the WHO criteria for severe malaria; 19 patients (54.3%) had more than one WHO criteria; 6 patients (2.6%) were initially admitted to the intensive care unit (ICU) and 5 more patients were subsequently referred selleck inhibitor to the ICU (11 total patients requiring intensive care).

Four patients received exchange transfusion as adjunctive therapy; all patients recovered uneventfully, but those treated in ICU had longer hospital stay (median 16 d vs 4 d; p < 0.001). All patients, irrespective of the

infecting Plasmodium species were admitted to the hospital; drug regimens employed are reported in Table 1. In our case file, mefloquine, either alone (173, 59.4%) or in combination with other drugs (27, 9.3%), was the most frequently used drug. It was employed in the treatment of all four Plasmodium species: in 177 patients infected by P falciparum (77.6%), 14 with P vivax (29.2%), 1 with P malariae (100%), 1 with Evodiamine P ovale (11.1%), and 3 mixed infections. The analysis of tolerance included 254 patients, thus excluding those who where treated with more than one drug: 34 (19.5%) adverse events were reported in those treated with mefloquine, 29 (76%) in the quinine-treated patients, and 2 (4.7%) in those receiving chloroquine (Figure 1). Cinchonism was registered exclusively in patients treated with quinine; only one patient treated with mefloquine discontinued treatment due to intractable vomiting. Incorrect use of anti-malarial drugs occurred overall in 25 patients (8.6%) in our case file (Table 2); anti-malarial errors were recorded more frequently in patients affected by P vivax malaria (14/48, 29.1%) than in those with P falciparum malaria (9/229, 3.9%; p = 0.0001).

Here, we use an Escherichia coliΔnanT strain to characterize

the function of known and proposed bacterial sialic acid transporters. We discover that the STM1128 gene from Salmonella enterica serovar Typhimurium, which encodes a member of the sodium solute symporter family, is able to restore growth on sialic acid to the ΔnanT strain and is Epacadostat able to transport [14C]-sialic acid. Using the ΔnanT genetic background, we performed a direct in vivo comparison of the transport properties of the STM1128 protein with those of sialic acid transporters of the major facilitator superfamily and tripartite ATP-independent periplasmic families, E. coli NanT and Haemophilus influenzae SiaPQM, respectively. This revealed that both STM1128 and SiaPQM are sodium-dependent and, unlike SiaPQM, both STM1128 and NanT are reversible secondary carriers, demonstrating qualitative functional differences in the properties of sialic acid transporters

used by bacteria that colonize humans. Sialic acids are a family of related nine carbon sugar acids that play important roles in the biology Tofacitinib ic50 of a wide range of both eukaryotic and prokaryotic organisms (Schauer, 2004; Vimr et al., 2004). In mammals, sialic acids are a predominant feature on the surface of many cell types, and bacteria have evolved multiple mechanisms to exploit these host-derived sugars (Vimr et al., 2004; Severi et al., 2007). For example, Escherichia coli is able to grow on the most common sialic acid N-acetylneuraminic

acid (Neu5Ac) as a sole carbon and nitrogen source (Vimr & Troy, 1985), which is important for successful colonization of the mouse gut (Chang et al., 2004). Other bacteria such as Haemophilus influenzae use host-derived Neu5Ac in an immune evasion mechanism by adding it as a terminal component of their lipopolysaccharide (Bouchet et al., 2003). While some pathogens have evolved de Elongation factor 2 kinase novo biosynthesis pathways for Neu5Ac (Vimr et al., 2004), many bacteria rely on the acquisition of Neu5Ac from their environment and hence require high-affinity transport systems (Bouchet et al., 2003). The pioneering work of Vimr and colleagues led to the first molecular characterization of a bacterial Neu5Ac transporter, which was the NanT protein from E. coli (Vimr & Troy, 1985). This is a secondary transporter and a member of the major facilitator superfamily (MFS) (Martinez et al., 1995). Very recently, another MFS family member, distinct from NanT, has been implicated in sialic acid uptake in Bacteriodes fragilis (Brigham et al., 2009) and Tannerella forsythia (Thompson et al., 2009). We and others have characterized a tripartite ATP-independent periplasmic (TRAP) transporter for Neu5Ac from H. influenzae, SiaPQM, that is important for virulence (Allen et al., 2005; Severi et al., 2005; Mulligan et al., 2009).

Light-emitting diodes with

narrow spectral emissions or notch filters facilitate these investigations. Practicalities may force a reliance on incandescent and fluorescent lights but, because of their complex spectra, comparing light of different colors is more difficult. Illuminance measures suffice when wavelength per se is not a central focus. The photosensitivity of a physiological or behavioral response to light depends on what is being measured. This is important as the photosensitivity of one response cannot be generalized to other functions. As an example, measurements were made of the thresholds of entrainment of wheel-running DNA Damage inhibitor rhythms at three wavelengths, and these were compared with the thresholds of two other non-image-forming visual system functions, i.e. masking and the pupillary light reflex. Dim light that entrained mice failed to elicit either masking or pupillary light reflex; in general, circadian entrainment is more sensitive by 1–2 log units than other measures of the non-image-forming visual system. In an artificial photic environment, dim light can entrain circadian rhythms even when it fails to produce more easily measurable acute responses to light such as phase shifting and melatonin suppression (Butler & Silver, 2011). As mentioned previously, not only does the

circadian system influence feeding and metabolism, but food cues can also act to entrain circadian rhythms (Saper, 2006; Patton & Mistlberger, 2013). If food presentation is restricted to Niclosamide a short temporal window (typically a few hours), animals

exhibit increased Y-27632 in vitro activity in anticipation of feeding [food anticipatory activity (FAA)]. Because this synchronization of behavior with feeding persists in the absence of the SCN, a separate designation of the food entrainable oscillator was coined (Stephan et al., 1979). The identification of the neural locus of the food-entrainable oscillator has been challenging. The dorsomedial nucleus of the hypothalamus (DMH) probably plays a role in food entrainment (Gooley et al., 2006; Fuller et al., 2008), although mice and rats can entrain to food cues in the absence of a DMH (Landry et al., 2006, 2007; Acosta-Galvan et al., 2011). In mice, DMH lesions lead to reduced FAA, whereas lesions of both the SCN and DMH result in enhanced FAA (Acosta-Galvan et al., 2011). These findings suggest that the DMH participates in FAA, but is not the sole neural locus of the food-entrainable oscillator. It is likely that metabolic cues from the periphery, communicated to the central nervous system, participate in food entrainment. For example, ghrelin cells in the stomach that signal hunger express clock genes, ghrelin administration leads to increased activity in animals fed ad libitum, and ghrelin and clock gene rhythms in these stomach cells are synchronized to feeding (LeSauter et al., 2009). Consistent with these findings, FAA is greatly reduced in ghrelin receptor knockout mice (Blum et al., 2009; LeSauter et al.

During the period, 185 children (122 families) attending the center for pre-travel advice agreed

to participate. One hundred sixty-seven (90%) children (109 families) were evaluated by the post-travel questionnaire. Three (2%) children had cancelled their journey and 15 (8%) this website were unobtainable for follow-up. Sex ratio was 1.0 and mean age 68 (SD = 54) months. Ninety-nine (54%) children traveled to Africa, 48 (26%) to Indian Ocean, 18 (10%) to Asia, and 20 (11%) to South America. The five most visited countries were the Comoros (22%), Senegal (18%), Kenya (8%), Cameroon (7%), and French Guyana (5%). The mean duration of travel was 29 days (SD = 19). One hundred eighty-three (99%) children were born in France, but only 103 (56%) had European maternal ascendance. Thirty-seven (20%) of the children lived with only one of the parents (monoparental families) and 41 (22%) children had state health insurance. One hundred two children (55%) were VFR and 83 (45%) were traveling for tourism. As shown in Table 1, VFR children significantly differed from tourists in age (younger), maternal origins (outside Europe), family structure (monoparental), health insurance (state insurance), siblings (higher number), destination (Indian Ocean), housing during travel (local housing), duration

of the stay (longer), and time Galunisertib nmr between pre-travel visit and departure (shorter). Table 2 reports the compliance with prophylactic measures among the 167 post-travel evaluated children. Only 75 (41%) children were already fully

immunized with routine vaccines.[7] Differences were observed in vaccine coverage: 84% for diphtheria, tetanus, poliomyelitis, pertussis, or Haemophilus influenzae type B, but IMP dehydrogenase 54% for hepatitis B. A routine vaccine update and travel-specific vaccines were proposed to 74 (40%) and 132 (71%) children, respectively. Among the 167 children for whom vaccination was recommended, 118 (71%) were fully compliant. Yellow fever vaccine was accepted in 100% of cases. Acceptance rates of hepatitis A, typhoid fever, and Bacillus Calmette Guérin immunizations were 75, 77, and 36%, respectively. Parents’ reasons for not going ahead with prescribed vaccinations (49 children) were: cost of vaccines (12%), fear of adverse events (12%), neglect of vaccination (6%), perceived inefficacy of vaccines (4%), or lack of time before departure (2%). One hundred sixty-one (87%) children were prescribed antimalarials: atovaquone-proguanil (46%), mefloquine (40%), doxycycline (9%), chloroquine (2%), and chloroquine plus proguanil (2%). Of those children 147 (91%) were evaluated on their return. All had used at least one form of protection against arthropod bites (repellent 95%, bed net 71%, or insecticides 54%) but only 46 (31%) children had used the three types of protection. The chemoprophylaxis was purchased for 136 (93%) children.