A key enzyme, most commonly an NRPS or polyketide synthase, produces a precursor molecule, which is subsequently modified by other enzymes encoded by the cluster. These genes usually produce a single product of small molecular weight, for example polyketides (lovastatin and aflatoxin B1), nonribosomal peptides (penicillin G and gliotoxin),

terpenes (gibbererellin) and indole alkaloids (fumitromorgin C), which are dispensable for cellular growth and have a restricted taxanomic distribution (Keller et al., 2005). The well-documented cytotoxic and phytotoxic properties of many of these compounds have long identified them as putative virulence factors. Gene expression profiling and candidate gene analysis of multiple SGI-1776 price secondary metabolite-producing species present us with the first opportunity to assess their role as a common molecular feature used by fungi to overcome universal challenges encountered in the host niche. Other secondary metabolites have impacts on virulence that are unique to

both the host environment and the stage of infection. The immunotoxic dipeptide gliotoxin, produced by a cluster of 19 genes in A. fumigatus (Cramer et al., 2006), is induced 14-h postinfection relative to laboratory culture (McDonagh et al., 2008) and is a virulence factor in a hydrocortisone acetate-treated, but not neutropenic, murine infection model. The action of gliotoxin as a virulence factor Metabolism inhibitor in vivo is most likely due to action against neutrophils, which is supported by ex vivo cellular assays (Bok et al., 2006a), and has recently been substantiated as acting at the level of proapoptotic gene family members in a physiologically relevant context using hydrocortisone acetate-treated BAK knockout mice (Pardo et al., 2006). A unifying model to NADPH-cytochrome-c2 reductase explain the existence of fungal clusters is currently unavailable, but it seems clear that multiple evolutionary mechanisms may explain their origin. Currently, three main hypotheses have been proposed, and gene expression analysis

represents a useful tool for determining the relative contribution of these hypotheses to fungal gene clustering. Horizontal gene transfer (HGT) suggests that clusters may both originate and be maintained from selective pressure following HGT events. Gene clusters that encode an entire metabolic pathway or virulence factor are more likely to result in a phenotypic advantage to the recipient genome (Walton, 2000). The gene duplication, diversification and differential gene loss (DDL) hypothesis emphasizes the fundamental features of specific genomic regions as being the driving force behind clustering. Areas of microbial eukaryotes that are most subject to high genetic and genomic variability are the subtelomeres, located between the telomeric end of linear chromosomes and chromosome-specific sequences (Farman, 2007).

Prognosis is severe in children, especially in those with age less than 1 year or severe malnutrition.[1] Adult mortality rates are also high in immunocompromised patients.[3, 6] Conversely, elderly patients without underlying disease and young immune-competent

adults are much more likely to have a favorable outcome,[4, 5, 7] as observed in our two patients. Shigellosis, because of its severity, should always be treated, whether bacteremia is found or not. But the global increase in antibiotic resistance limits the choice of drugs.[1] Among Dasatinib order recommended treatments, fluoroquinolones or third-generation cephalosporins are the best choices for Shigella bacteremias, but sensitivity must be confirmed. Due to an absence of controlled studies, there is no consensus on treatment duration. Courses generally range between 5 and 14 days, depending on the severity and duration of symptoms.[3-5] Shigella bacteremia is fortunately uncommon in healthy travelers. When an underlying disease is absent, it should alert the physician to the possibility

of a transient co-morbid condition. These case reports underline the importance for travelers to seek pre-travel advice and be prepared to prompt self-treatment of diarrhea with an antibiotic-containing regimen. The authors thank Dr M. Nesemann for linguistic assistance. The authors state they have no conflicts of interest to declare. “
“Clinical and laboratory findings are described from 77 persons from Nairobi, Kenya, of whom 66 Epigenetic inhibition acetylcholine were diagnosed with acute Schistosoma mansoni infection following a trip to Mwanza, Tanzania. Unusual ocular symptoms were observed as a rare manifestation of acute schistosomiasis. The outbreak highlights

the risk of swimming in Lake Victoria. In August 2008, the Seventh Day Adventist (SDA) group of churches in East Africa organized a family retreat to Mwanza in Tanzania, located on the shores of Lake Victoria. They were there for several days, during which most of them swam in the lake, having been assured that the water was “safe.” Once the retreat was over, the families returned to their homes all over the East African region and beyond. Approximately 8 weeks after exposure to the lake water, on October 28, 2008, a 10-year-old girl was referred to the Centre for Tropical and Travel Medicine (CTTM) laboratory for malaria and hemoglobin testing. The child presented with general malaise which her mother thought was malaria, but she also had periorbital edema. On examination of the Giemsa-stained blood slide, marked eosinophilia was noted in the absence of malaria. Given the history of recent swimming in Lake Victoria during the church retreat to Mwanza, a blood test for bilharzia antibody was requested, which was positive at a titer of 1 : 1024. The child was put on treatment with praziquantel at a dose of 40 mg/kg daily for 4 days, and her symptoms subsided rapidly.

e. a PI). There is no randomized comparison of these three strategies. However, in one study a lower number of emergent resistance mutations were seen in patients switching to a PI compared with those undertaking a simultaneous or staggered stop [54]. Therapeutic plasma concentrations of EFV can also be detected up to 3 weeks after stopping the drug in some

patients and thus a staggered stop of 1 week may potentially be inadequate to prevent emergence of NNRTI mutations [56]. The optimal duration of replacement with a PI is not known, but 4 weeks is probably advisable. Data on how to switch away from EFV to an alternative ‘third’ agent are either non-existent, or of low or very low quality. Based on pharmacological principles, there is little rationale for any strategy other than straightforward Selleck Y27632 substitution

when switching to a PI/r or RAL. Pharmacokinetic studies show that straightforward substitution with ETV and RPV may result in slightly lower concentrations of either drug for a short period following switching, but limited virological data suggest that risk of virological failure with this strategy is low. Different strategies for switching to NVP have been proposed, but no comparative data are available to guide the choice of strategy. Limited data suggest that the dose of MVC should be doubled in the week following switching (unless given together with a PI/r). If switching away from EFV is undertaken when VL is likely to still to be detectable (e.g. because http://www.selleckchem.com/products/r428.html of CNS intolerance within the first few weeks of starting EFV), substitution with a PI/r in preference to a within-class switch is advised. Switching a component of an ART regimen is frequently considered click here in patients to manage drug side effects or

address adherence issues. ARVs that either induce or inhibit drug-metabolizing enzymes have the potential to affect the plasma concentrations of the new agent. This applies in particular to switching away from NNRTIs. Induction of drug metabolizing enzymes by EFV is likely to persist for a period beyond drug cessation. Consideration should also be taken of whether or not VL is maximally suppressed when planning how to switch away from EFV to an alternative agent. Broadly, strategies for switching from EFV to an alternative ‘third’ agent may be summarized as follows. A pharmacokinetic study performed in HIV-positive individuals suggested that patients changing from EFV to NVP should commence on 200 mg twice a day to ensure therapeutic plasma concentrations and potentially avoid selection of resistance to NVP [57]. However, no patient in the NVP lead-in group experienced virological failure in the 3-month follow-up period.

coli and an isogenic mutant lacking relA, the gene coding for the ppGpp synthetase gene. There was no change in β-galactosidase activity (data not EX 527 in vitro shown). This suggests that the stringent response might not be involved in aah-aidA control. Other nutrient-limitation pathways might therefore be involved. In summary, we identified,

in a wild-type pathogenic strain of E. coli, two putative promoters upstream of aah likely to drive the production of aah-aidA bicistronic transcripts. Our work shows that aah and aidA are induced at the early-stationary phase, likely because of nutrient starvation. This pattern leads us to hypothesize that RpoS is the principal regulator of the expression of the aah-aidA operon, at least in the wild-type strain and the conditions we used. This hypothesis is consistent with the consensus sequences of one promoter we identified upstream of aah. Other promoters are likely to be present upstream of aah and aidA and could play further roles in conditions not reproduced in our assays. The preferential expression of AIDA-I at a high density of bacteria and/or during nutrient starvation is consistent with the fact that AIDA-I is mediating bacterial auto-aggregation.

It would indeed make sense to turn on the gene coding learn more for AIDA-I in an environment where a high density of bacteria are present or under adverse conditions of poor nutrient availability, so as to form ‘micro-colonies’ of bacteria of the same kind and increase Acetophenone survival chances. Similar effects of nutrient starvation influencing the expression of virulence genes

in pathogenic E. coli have been observed before: for instance, in enterohemorrhagic E. coli, the expression of LEE genes is activated in response to starvation and bacterial adherence is increased (Nakanishi et al., 2006), and in uropathogenic E. coli, the entry into the stationary phase triggers the expression of fimbriae and an increase in the frequency of adherent bacteria (Aberg et al., 2006). As we were writing this report, another study was published on the regulation of aah and aidA (Benz et al., 2010). This work was performed in a laboratory strain of E. coli with the aah-aidA region cloned on a multicopy plasmid. It therefore complements well our own study performed in a wild-type background. The two aah promoters we identified were also found in this recent study and experiments showed the existence of a bicistronic message, in agreement with our conclusions. Two additional promoters were found upstream of aidA. As explained above, it is possible that the presence of a multicopy plasmid and/or the background of a laboratory strain of E. coli allowed the identification of these promoters that we failed to find.

2,11,16 Travel to altitude could have more severe consequences for diabetic patients with complications or poor metabolic control, and they should be evaluated and counseled accordingly. All diabetic patients should be carefully screened for complications that could increase their risk associated with exercise or exposure to altitude.11 The Web site www.mountain-mad.org is an excellent resource for people with diabetes who are interested in mountain pursuits.84 Ri-Li and colleagues found that obese people had worse AMS scores than non-obese counterparts

at a simulated altitude of 3,658 m.85 This effect is attributed to nocturnal desaturation associated with periodic, apneic breathing.85,86 Furthermore, excess abdominal weight increases the likelihood of OSA and obesity–hypoventilation selleck chemicals syndrome.8 These factors can exacerbate both hypoxemia and pulmonary hypertension which may increase an individual’s risk for

developing HAPE.8,43 Excess body weight may also complicate or preclude stretcher rescue from remote locations. Obesity–hypoventilation syndrome is a contraindication to high altitude travel. If such travel is necessary, supplemental oxygen and prophylactic acetazolamide are recommended.8 The effect of altitude on the seizure threshold has not been studied in depth. However, many well-controlled epileptics safely travel to altitude and are at no known increased risk Decitabine manufacturer for development of altitude-related illness or seizures.43,87 buy RGFP966 There have been multiple case reports of seizures occurring in non-epileptic individuals at altitude, including one fatal case.12,87–91 Daleau and colleagues reported a case where previously undiagnosed hyperventilation-induced

seizures were unmasked in a patient with a positive family history for epilepsy.92 Basnyat also reported a single case of grand mal seizures at high altitude in a well-controlled epileptic patient on anticonvulsant medications.87 Seizures at high altitude are believed to be provoked by a number of potential factors including respiratory alkalosis, hypocapnia, hypoxia, or sleep deprivation.12,87 Fluoroquinolone antibiotics prescribed for gastroenteritis have also been implicated in two case reports87,88 because of their potential for lowering the seizure threshold.93 Lastly, although the potential for having a seizure may not be greatly elevated at altitude, consideration must be given to the additional potential for harm, should a seizure occur in a remote location or while performing high risk technical mountaineering maneuvers. The risk of stroke at altitude may be increased due to hyperviscosity secondary to polycythemia, dehydration, cold exposure, and forced inactivity. Ischemic stroke and cerebral artery thrombosis are potential complications of high altitude cerebral edema.

rpoA could be a useful marker for identifying and classifying S. pneumoniae, selleck compound S. mitis, and S. oralis from closely related taxa. Family Streptococcaceae encompasses a broad range of gram-positive, catalase-negative, chain-forming coccus-shaped organisms. Currently, 92 species are recognized, many of which are associated with disease in humans and animals (http://www.bacterio.cict.fr). Among these group species, Streptococcus pneumoniae, the most common cause of pneumonia, bacterial meningitis, and nongonococcal urethritis in humans (Marrie et al., 1989; Hall et al., 1995; Fine et al., 1996), is frequently detected in the oral environment.

By contrast, two viridans group streptococci, Streptococcus mitis and Streptococcus oralis, which constitute major populations on oral soft tissues, cause dental caries and endocarditis (Willcox et al., 1988; Dyson et al., 1999). Precise discrimination

among the strains is essential for accurate diagnosis and treatment. Identification and classification of these organisms has long been considered difficult, however, because Nintedanib ic50 they have a close, common genetic ancestry (Whiley & Beighton, 1998; Whatmore et al., 2000; Mager et al., 2003). In recent years, molecular genetic analyses based on the 16S rRNA gene have provided new insights into the phylogenetic inter-relationships of many organisms (Bentley et al., 1991) and provided a powerful means for characterizing the level of species (Stackebrandt selleck chemicals et al., 1991; Fox et al, 1992; Stackebrandt & Goebel, 1994). However, the 16S rRNA gene molecule from members of closely related species may be so conserved that it cannot be used to distinguish between strains at the species level (Stackebrandt et al., 2002). Indeed, the nucleotide sequences of the 16S rRNA genes from S. mitis and S. oralis are almost (>99%) identical to that of S. pneumoniae, making the use of 16S rRNA gene alone insufficient for discrimination among these species (Suzuki et al., 2005). Housekeeping protein-coding genes are thought to

evolve faster than rRNA genes, and have been proposed as suitable phylogenetic markers for the identification and classification of bacteria (Palys et al., 1997, 2000). The aim of this study was to focus on the evaluation of the rpoA (RNA polymerase α subunit) gene for its reliability and usefulness as a new marker for discrimination among Streptococcus species. The 28 bacterial strains used in this study are listed in Table 1 and were obtained from the Korea Collection for Type Culture (KCTC, Daejeon, Korea), Culture Collection of Antibiotics Resistant Microbe (CCARM, Seoul, Korea), Korean Collection for Oral Microbiology (KCOM, Gwangju, Korea), Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig Germany), and the American Type Culture Collection (ATCC, Manassas, VA). Each bacterial strain was grown on sheep blood agar plates (Asan Pharm Co.

This pattern of anatomical connectivity was confirmed with RSFC in the see more human brain and clearly sets ventral area 6 apart from areas 44 and 45. As can be seen in Fig. 1, the functional connectivity of area 6 was restricted to the anterior

part of the supramarginal gyrus that is delimited by the posterior ascending ramus of the Sylvian fissure. The pattern of RSFC associated with Cluster 3 (Fig. 5) supports this conclusion, which was also confirmed by the direct contrasts between BA 6, and BAs 44 and 45 (as shown in Fig. 1 and Table 1). The strong RSFC of BA 6 with the most anterior part of the inferior parietal lobule and the absence of correlations with the posterior part of the supramarginal gyrus and the angular gyrus define a unique profile of parietal RSFC for ventral BA 6. By contrast, areas 44 and 45 exhibited a functional connectivity pattern with the posterior supramarginal gyrus and the angular gyrus (Fig. 1), consistent with predictions Linsitinib in vitro from the macaque monkey studies (Petrides & Pandya, 2009). Furthermore, areas 44 and 45 had strong correlations with the cortex in the superior temporal sulcus and the temporal cortex just below it, namely the middle temporal gyrus (Figs 1 and 2). The strong distinction between the connectivity patterns associated with ventral area 6 relative to areas 44 and 45 is most evident in the results of the clustering analysis. The simplest

and most robust partitioning of the data (K = 2, see Fig. 3) was one that separated ventral area 6 into one cluster, and areas 44, 45 and the rest of the inferior frontal gyrus into another (see top row of Fig. 4). The clear separation DAPT price between ventral area 6 and area 44 anteriorly was also present for the optimal solution (K = 4, see Fig. 4). In both monkey and human brains, ventral area 6 is a typical premotor cortex that lacks layer IV, whereas area 45 is a typical prefrontal

cortex with a well-developed layer IV (Brodmann, 1909; Amunts et al., 1999; Petrides & Pandya, 2002). Area 44, which lies between areas 6 and 45, does possess a layer IV, but it is interrupted and not well developed. Consequently, there has long been confusion as to whether BA 44 should be considered a premotor zone that is functionally similar to premotor cortex or whether BA 44 is functionally more similar to prefrontal BA 45. For instance, some investigators have considered Broca’s region to include both BAs 44 and 45 (Amunts et al., 1999) while others have restricted it to BA 44 (Mohr et al., 1978). The present results address this issue. The functional connectivity patterns of BAs 44 and 45, which together comprise Broca’s area, were more similar to one another than to the RSFC of ventral BA 6. This conclusion is also consistent with a study by Amunts & Zilles (2006), who examined the architectonic and neurochemical profiles of BA 44 and concluded that it shares more features with BA 45 than with BA 6.

The transplanted forebrain cells failed to activate regulatory genes specific of cerebellar interneurons, such as Pax-2 (Maricich & Herrup, 1999). Nonetheless, they engrafted in the cerebellum and developed mature neurons, which were assigned SB431542 to different categories

of local interneurons, based on their morphology and localization. Hence, it was concluded that extracerebellar donors differentiate into cerebellar-like interneurons. In the article published in this issue of EJN, Rolando et al. (2010) compared the developmental potentialities of progenitors from different sites along the neuraxis exposed to the postnatal cerebellar PWM. To identify the phenotypes acquired by donor cells, these investigators applied a set of concurrent criteria, including expression of region-specific transcription factors, morphological features, GKT137831 cost neurochemical profiles and position in the recipient architecture. Most importantly, starting from the recent work of Fernando Rossi and collaborators, showing that the phenotype and position of cerebellar

interneurons are specified according to precise spatio-temporal patterns (Jankovski et al., 1996; Carletti et al., 2002; Leto et al., 2006, 2009), Rolando et al. (2010) asked whether extracerebellar donors shared the same developmental phases and final fate of the cerebellar interneurons generated at the age when transplantation was done. Although the results of these experiments are partly consistent with those of Milosevic et al. (2008), the conclusions

are quite different. The morphology, position and expression of type-specific markers in donor neurons did not correspond to those of their age-matched cerebellar counterparts. Furthermore, the morphological features of donor neurons that may be termed ‘cerebellar-like’ appeared to result from local interactions at the homing site rather than from the unfolding of a host-specific ontogenetic program. Interestingly, the acquisition of such features occurs more frequently when donor cells are derived from sites close to the cerebellum along the rostro-caudal extent of the neuraxis. Thus, although exogenous neurons stably engraft in the cerebellum and acquire some features reminiscent of local interneurons, it is clear that they develop according Cyclooxygenase (COX) to their own native properties and fail to become integrated into the host ontogenetic mechanisms. Thus, the results reported by Rolando et al. (2010) indicate that changing the regional identity of neural progenitors is not an easy task. “
“Synaptic transmission is a complex process comprised of several steps. These include the arrival of action potentials at presynaptic terminals, the activation of presynaptic Ca2+ channels, the binding of Ca2+ ions to the sensors of exocytosis, the fusion of synaptic vesicles with the presynaptic plasma membrane, the release of transmitter into the synaptic cleft, and ultimately the activation of postsynaptic receptors.

Q-Sepharose, Phenyl Superose, ECL Western blotting detection reagents were obtained from Amersham. All other biochemicals were of the highest grade available. The WHO reference strain of L. donovani (MHOM/IN/80/Dd8) was obtained from Imperial College London (UK) and maintained in vitro as promastigotes in RPMI 1640, supplemented with 10% heat-inactivated fetal bovine serum containing 40 μg mL−1 gentamycin at 25 °C. The TA cloning vector pGEM-T Easy was used to clone the PCR product, and the pET-28(a) vector having both N and C terminal His6-tag was used for expression of the recombinant protein. The recombinant plasmids were transformed into E. coli BL21 (DE3) cells for expression. PCR primers 5′-CATATGGGGTTCTTCTCGGATTCGGTAG-3′

(forward) and 5′-AAGCTTCGCGTGGCCGGCAATCTCCTTG-3′ (reverse) with NdeI restriction INK 128 in vivo site at the forward and HindIII at the reverse end shown as underlined were designed based on the L. major SSN gene (U30455) sequence. Amplification of the SSN gene was carried out using genomic DNA of L. donovani as template. The reaction mixture contained 50 ng of genomic DNA, 1.5 U Taq polymerase, 0.5 μM primer (each) and 200 μM each dNTPs in 50 μL PCR reaction mixture volume. After initial denaturation at 94 °C

for 3 min, the PCR reaction was programmed for 30 selleck inhibitor cycles, with each cycle including denaturation at 94 °C for 30 s, 50 °C annealing temperature for 1 min and extension at 72 °C for 2 min. There was a final extension at 72 °C for 10 min. The amplified product after gel purification was cloned in the pGEM-T-Easy vector and transformed in E. coli DH5α competent cells. Nucleotide sequencing of the recombinant construct was carried out in both directions to confirm the sequence of the amplicon and the sequence was submitted to NCBI GenBank. The recombinant construct was digested with

restriction endonuclease NdeI and HindIII and subcloned into the prokaryotic expression vector pET-28(a) for overexpression and purification of recombinant LdSSN. SSN genes of diverse species at the level of the deduced amino acid sequence from Swiss prot or cAMP protein data bank (http://www.expasy.org/sprot/) were aligned with clustalw, followed by the generation of a phylogenetic tree. The recombinant construct pET-28 (a)-LdSSN was used to transform competent E. coli BL21 (DE3). The plasmid was grown overnight as primary culture. Luria–Bertani broth (1 L) containing 25 μg mL−1 kanamycin was reinoculated at 0.1% cell density and grown at 37 °C under constant shaking. The culture was induced by 0.1 mM isopropyl-β-d-thiogalactopyranoside (IPTG) (OD600 nm 0.4) and incubated at 20 °C for another 12 h under gentle shaking at 120 r.p.m. Uninduced culture was taken out and run as a negative control. The overnight grown culture was harvested at 5000 g for 15 min at 4 °C and suspended in buffer A [20 mM Tris buffer, pH 7.

Furthermore, in the subset of Cabozantinib mw patients with an eGFR pre-cART ≥90 mL/min per 1.73 m2, the time of a confirmed eGFR reduction from pre-cART levels was alternatively

defined as the date of the first of two consecutive eGFR values <90 mL/min per 1.73 m2. Poisson regression analyses including the same variables as were included in the main analysis were employed to identify independent predictors of a reduction of eGFR. Patients included in the analysis (n=1505) showed significant differences in immunovirological variables compared with excluded patients (n=5762; Table 1); included patients had higher CD4 cell counts (505 vs. 450 cells/μL, respectively; P<0.0001) and higher median HIV RNA levels (4.14 vs. 3.00 log10 HIV-1 RNA copies/mL, respectively; P<0.0001) at baseline. Included patients were younger (38 vs. 39 years, respectively; P<0.0001) and more likely

to be affected by diabetes and/or hypertension (2%vs. 1%, respectively; P=0.02); a lower percentage of included patients acquired HIV infection thorough injecting drug use (29% of the included patients vs. 35% of the excluded patients). There were no clinical differences in the percentage of female patients or CD8 cell count. A total of 1505 patients satisfied the inclusion criteria for the cross-sectional analysis. The clinical and immunovirologic characteristics of the patients, stratified by eGFR at baseline selleck (<90 or ≥90 mL/min/1.73 m2), are summarized in Table 2. A

confirmed eGFR<90 mL/min/1.73 m2 was observed in 363 (24%) of the patients. Of these, 353 (97%) had an eGFR in the range of 60–89 mL/min/1.73 m2, while only 10 patients Histamine H2 receptor (3%) had an eGFR of 30–59 mL/min/1.73 m2 and none had an eGFR below 30 mL/min/1.73 m2. In univariable analysis, compared with patients with normal eGFR, patients with a value of eGFR<90 mL/min/1.73 m2 at baseline were older, had higher CD4 cell counts, and were more likely to be female and to have suffered from diabetes and/or hypertension prior to baseline; in contrast, patients with normal eGFR were more likely to be coinfected with hepatitis B or C virus (Table 2). After adjustment, older age [odds ratio (OR) 1.58 per 10 years older; 95% confidence interval (CI) 1.37–1.82], female gender (OR 2.41 vs. male; 95% CI 1.75–3.31), a prior history of diabetes and/or hypertension (OR 2.36 vs. neither; 95% CI 1.08–5.14), baseline CD4 count (OR 1.06 per 100 cells/μL higher; 95% CI 1.01–1.11) and hepatitis coinfection (OR 0.51 vs. HIV monoinfection; 95% CI 0.34–0.78) were the sole independent predictors of a value<90 mL/min/1.73 m2 at baseline (Table 2). A total of 644 patients (43% of the total studied) started cART at some point during follow-up and were included in the longitudinal analysis (Table 3). The median calendar year of cART initiation was 2005 (range 2000–2009) and the median number of creatinine values post cART was 6 [interquartile range (IQR) 2–10].