Urgent referral would allow confirmation of a diagnosis of HIV in

Urgent referral would allow confirmation of a diagnosis of HIV in an infant and treatment to prevent disease progression. Individual feedback was sent to the units who sent guidelines, check details to allow them to improve their guidelines. Two units asked for a template to produce local guidelines. In summary, mother-to-child transmission of HIV is preventable. All maternity units should have

local guidelines, based on the BHIVA/CHIVA pregnancy guidelines, to allow them to manage infants born to HIV-infected women. Other regions should review local guidelines to ensure that they give enough information to manage both low-risk and high-risk infants, together with information on how and when to seek expert advice. “
“Quantification of obligate biotrophic parasites has been a long-standing problem in plant pathology. Many attempts have been made to determine how much of a pathogen is present in infected plant tissue. Methods of quantification

Trametinib included scoring disease symptoms, microscopic evaluation, determination of specific compounds like Ergosterol, and lately nucleic acid-based technologies. All of these methods have their drawbacks, and even real-time PCR may not be quantitative if for example the organism of interest has specific and differing numbers of nuclei in different infection structures. We applied reverse transcription (RT) real-time PCR to quantify Uromyces fabae within its host plant Vicia faba. We used three different genes, which have been shown to be constitutively expressed. Our analyses show an exponential increase of fungal material between 4 and 9 days post inoculation and thereafter reaching a steady state of around 45% of total RNA. We also used haustorium-specific genes to determine the amount

of haustoria present at each time point. These analyses parallel the development of the whole fungus with the exception of the steady-state level, which is only around 5% of the total RNA. This indicates that RT real-time PCR is a suitable method for quantification of obligate biotrophic parasites, and also for the differentiation of developmental stages. All higher organisms exhibit a more or less pronounced association with a plethora of symbiotic microorganisms, some of them beneficial, some of them neutral, and some of them pathogenic. While the determination second of the number of mutualistic or neutral symbionts has more of an academic value, accurate quantification of pathogen abundance is a critical issue in medicine and plant pathology. There have been numerous approaches to quantify the number of pathogens present in various host–parasite interactions at any given time point of pathogenesis. Traditionally, visual inspection and scoring of disease symptoms have been used to determine disease severity (Pei et al., 2002; Bock et al., 2008). Lately, this type of rating has been complemented by digital image analysis (Bock et al., 2008).

azotoformans were reported This investigation adds to the organi

azotoformans were reported. This investigation adds to the organizational Y-27632 order pattern diversity of the carotenogenesis gene cluster and the variety of CrtI in photosynthetic bacteria. The results of the

present study may provide a new gene resource for the reconstruction of carotenogenesis pathways to produce engineered carotenoids. Photosynthetic bacterial CGMCC 6086 was isolated from domestic sewage in Xiaoqinghe, Jinan, Shandong province. It was grown semianaerobically under phototrophic conditions at 30 °C for 48 h in RCVBN medium (Weaver et al., 1975). Escherichia coli strain BL21 (DE3) was used as the expression host and was grown aerobically at 37 °C in LB medium. Antibiotics were added to LB medium to a final concentration of 50 μg mL−1 (ampicillin) or 25 μg mL−1 (chloramphenicol), if required. Bacterial CGMCC 6086 was identified through morphological features, carotenoid composition, utilization of electron donors and carbon sources, and 16S rRNA gene sequence. Carotenoids in CGMCC 6086 were extracted and analyzed using the method described later. Utilization of electron donors and carbon sources were tested in RCVBN medium by replacing malic acid with organic acids, sugars, or alcohols in anaerobic

light or anaerobic dark denitrifying conditions. 16S rRNA gene was amplified through PCR using the universal primers 27f and 1525r (Table 1). Genomic DNA was extracted using a Biospin bacterial genomic DNA extraction kit (BioFlux, Japan), and PCR was performed using Taq DNA polymerase (TaKaRa, Japan). Sequence analysis RGFP966 nmr was performed using the nucleotide blast program (http://www.ncbi.nlm.nih.gov/BLAST/). AGAGTTTGATC MTGGCTCAG AAGGAGGTGA TCCAGCC CGCCCATTCCGG GCAATCCT GGCGCCCATATCA GCGCGAAA ACCCGGTCGCCCG GCTTGAA GGCGCTGCACCACG CGGGCAA GCCGCAAAGAGAAC GCCTGA Sequence between the crtAIB-tspO and crtCDEF fragments GCCCCGAAGCCCGG GCCTGA GGCCTTCGGACGC CTCCTGA GCCGGCTGGCGCTTT CCCAA TGCCATATGCCCGC

GACCAAGCATGT GTCAAGCTTTTCCGC GGCCAGCCTTT GTAGGATCCGATGAC GGTCTGCGCAAAAA TGCGAGCTCTTAACTG ACGGCAGCGAGT TGCCATATGAATAATC CGTCGTTACTC TAAGGTACCCTAGAGC GGGCGCTGCCAGA The carotenogenesis gene cluster of Rba. azotoformans CGMCC 6086 was cloned via PCR amplification. All the PCR primers are listed in Table 1. for Primers Ra-Ad, Ra-Fd, Ra-Od, and Ra-Cd were designed based on reported sequence of carotenogenesis gene clusters in Rba. sphaeroides (GenBank accession nos. CP001150, CP000577, CP000661, AF195122, and AJ010302). PCR was performed using LA Taq DNA polymerase and GC buffer I (TaKaRa). The genomic DNA of Rba. azotoformans CGMCC 6086 was used as the template. The amplification fragments were inserted into the pMD18-T vector (TaKaRa) and sequenced. Sequence alignments were performed with the protein blast program (http://www.ncbi.nlm.nih.gov/BLAST/).

Alternatively, cannabinoid-mediated spinal analgesia might be eli

Alternatively, cannabinoid-mediated spinal analgesia might be elicited through completely different mechanisms. Hegyi et al. (2009) showed that CB1 receptors in the spinal cord dorsal horn are not only found on neurons but also on half of the astrocytes and on the majority of microglia cells. Both types of glia cells contribute to pathological pain syndromes (Miraucourt et al., 2007; Inoue & Tsuda, 2009) and a CB1 receptor-dependent regulation of these cells might very well contribute to cannabinoid-mediated spinal analgesia. Regardless of the eventual explanation for these discrepant results, increasing evidence indicates that the action

cannabinoids and CB1 Napabucasin purchase receptors in vivo is more complex than apparent ex-vivo. The study by Zhang et al. (2010) will certainly not remain the last surprise in cannabinoid research. “
“Peripheral nerve injury induces axonal degeneration and demyelination, which are collectively referred to as Wallerian degeneration. It is generally assumed that axonal degeneration is a trigger for the subsequent demyelination processes such as myelin destruction

and de-differentiation of Schwann cells, but the detailed sequence of events that occurs during this initial phase of demyelination following axonal degeneration remains unclear. Here we performed a morphological analysis of injured sciatic nerves of wlds mice, a naturally occurring mutant selleck screening library mouse in which Wallerian degeneration shows a significant delay. The slow Wallerian degerenation phenotype of the wlds mutant mice would enable us to dissect the

events that take place during the initial phase of demyelination. Ultrastrucural analysis using electron microscopy showed that the initial process of myelin destruction was activated in injured nerves of wlds mice even though they exhibit morphologically complete protection of axons against nerve injury. We also found that some intact axons were completely demyelinated in degenerating MycoClean Mycoplasma Removal Kit nerves of wlds mice. Furthermore, we observed that de-differentiation of myelinating Schwann cells gradually proceeded even though the axons remained morphologically intact. These data suggest that initiation and progression of demyelination in injured peripheral nerves is, at least in part, independent of axonal degeneration. “
“Evaluation of the behavioral ‘costs’, such as effort expenditure relative to the benefits of obtaining reward, is a major determinant of goal-directed action. Neuroimaging evidence suggests that the human medial orbitofrontal cortex (mOFC) is involved in this calculation and thereby guides goal-directed and choice behavior, but this region’s functional significance in rodents is unknown despite extensive work characterizing the role of the lateral OFC in cue-related response inhibition processes.


“Recently, travel to underdeveloped and exotic destination


“Recently, travel to underdeveloped and exotic destinations has increased substantially. International travel is a multi-billion dollar industry exceeding $900 billion US IWR-1 datasheet dollars (600 euros) in 2008. By the year 2020, it is expected

that the number of international travelers will exceed 1 billion, half being for leisure purposes and approximately 15% business related.1Prior to departure for travel, it is widely recommended to consult with a specialist in travel health, as many travelers are unaware of the immunizations and preventative measures that are recommended. Pharmacists are accessible healthcare professionals who have unique opportunities to provide education and administration of immunizations FK228 chemical structure to this population. Over the past two decades, pharmacists have become more involved in the provision of travel medicine services in a variety of settings.2–5 The Clinical Pharmacy

International Travel Clinic (CPITC), established in the early 1990s, is a telepharmacy consultation service run by pharmacists from the Kaiser Permanente Colorado region.2 The team, composed of five clinical pharmacists, a pharmacy technician, and a consulting infectious diseases physician, provide phone consultation for approximately 9500 travel patients every year, following referrals from primary care physicians (PCPs) or customer service associates. As no appointments are required, patients receive their consultation at the time they call the service. The pharmacists provide recommendations regarding travel immunizations, medications, and preventive measures against diseases abroad, and they attain prescriber co-signatures for these orders. It is estimated that the CPITC pharmacists could save $47,000 per year in unnecessary immunizations with this consultation service.3 Community pharmacists have also become involved in travel medicine services, due to their ease of accessibility with many convenient locations, long hours of operation, and the ability to immunize.3,5 One pretravel health program, TravelRx, offered by a supermarket

chain pharmacy in Central Virginia, provides initial phone consultation followed by individualized appointments in a private counseling room within the pharmacy for approximately 1000 patients per year.4Following the patient interview and assessment of travel-related needs, the patient’s Acyl CoA dehydrogenase PCP is contacted to gain authorization for the administration of immunizations and medications; the pharmacist then schedules an appointment for the patient’s travel education and immunizations. Following the patient’s visit, the pharmacist follows up with both the physician (to provide documentation of the patient’s immunizations) and the patient (to complete any additional vaccine series post-travel). Patients expressed a high level of satisfaction with the pharmacist-run program through patient satisfaction surveys, although no outcomes were formally assessed.

, 2005) By shifting the position of the reference objects relati

, 2005). By shifting the position of the reference objects relative to the gaze fixation target, it was possible to determine whether neurons represented the position of missing components in viewer-centered

or object-centered spatial coordinates. If neurons coded the position of missing object components in viewer-centered coordinates, their activity would vary as a function of whether they were located to the left or right of the gaze fixation target (at the midline of viewer-centered frames of learn more reference). If, instead, neurons coded the position of missing object components in object-centered coordinates, their activity would vary as a function of whether components were missing from the left or right side of the copy object, relative to its intrinsic midline. A population of neurons in area 7a was found which represented the position of missing components in object-centered coordinates, relative to the midline of the object (Chafee et al., 2007). Kinase Inhibitor Library These neurons were similarly activated during the copy period whenever the missing component was located on a preferred side of the copy object, regardless of where the copy object was presented in viewer-centered space (Fig. 7A). The above

data provided evidence that the parietal neurons supported a spatial cognitive process during the object construction task that analyzed object structure and that represented the results of the analysis in object-centered coordinates. However, parietal neurons were heterogeneous in terms of the spatial coordinate

system they used to represent space, and neurons coding position in viewer- and object-centered position were both present in area 7a (Chafee et al., 2007). A decoding analysis quantified the information carried by the activity of the two simultaneously active populations in their respective coordinate systems, and found that variation in viewer-centered information preceded and could predict variation in object-centered Diflunisal information over time within a trial (Crowe et al., 2008). This observation was consistent with the hypothesis that spatial information provided by the visual input coding position, initially in viewer-centered (retinocentric) coordinates, was converted into spatial information coding position in object-centered coordinates over time, and that a correlate of this transform could be detected in parietal cortex. The above neural data provide evidence that parietal neurons encode spatial information that is the product of a spatial cognitive analysis applied to the visual input to meet a specific behavioural objective. That functional conclusion was further substantiated by the results of neurophysiological recordings in parietal area 7a of monkeys performing a visual maze task (Fig. 8).

We also measured performance on an effort-based discounting task

We also measured performance on an effort-based discounting task. We then assessed D1 and D2 dopamine receptor mRNA expression in subregions of the prefrontal cortex and nucleus accumbens using in situ hybridisation, and compared these data with behavioral

performance. Expression of D1 and D2 receptor mRNA in distinct brain regions was predictive of impulsive action. A dissociation within the nucleus accumbens was observed between subregions and receptor subtypes; higher D1 mRNA expression in the shell predicted greater impulsive action, whereas lower D2 mRNA expression in the core predicted greater impulsive action. click here We also observed a negative correlation between impulsive action and D2 mRNA expression in the prelimbic cortex. Interestingly, a similar relationship was present between impulsive choice and prelimbic cortex D2 mRNA, despite the fact that behavioral indices of impulsive action and impulsive choice were uncorrelated. Finally, we found that both high D1 mRNA expression in the insular

cortex and low D2 mRNA expression in the infralimbic cortex were associated with willingness to exert effort for rewards. Notably, dopamine receptor mRNA in these regions was not associated with either facet of impulsivity. Kinase Inhibitor Library mouse The data presented here provide novel molecular and neuroanatomical distinctions between different forms of impulsivity, as well as effort-based decision-making. “
“Polyphenol resveratrol (RSV) has been associated with Silent Information Regulator T1 (SIRT1) and AMP-activated protein kinase (AMPK) metabolic stress sensors and probably responds to the intracellular energy status. Our aim here was to investigate the neuroprotective effects of RSV and its association with SIRT1 and AMPK signaling in recurrent ischemia models. In this study, elderly male Wistar rats received

a combination of two mild transient middle cerebral artery occlusions (tMCAOs) as an in vivo recurrent ischemic model. Primary cultured cortical neuronal cells subjected to combined oxygen–glucose Diflunisal deprivation (OGD) were used as an in vitro recurrent ischemic model. RSV administration significantly reduced infarct volumes, improved behavioral deficits and protected neuronal cells from cell death in recurrent ischemic stroke models in vivo and in vitro. RSV treatments significantly increased the intracellular NAD+/NADH ratio, AMPK and SIRT1 activities, decreased energy assumption and restored cell energy ATP level. SIRT1 and AMPK inhibitors and specific small interfering RNA (siRNA) for SIRT1 and AMPK significantly abrogated the neuroprotection induced by RSV. AMPK-siRNA and inhibitor decreased SIRT1 activities; however, SIRT1-siRNA and inhibitor had no impact on phospho-AMPK (p-AMPK) levels.

Self-treatment

Self-treatment learn more for probable malaria was always atovaquone/proguanil, according to French recommendations. The preferred regimens for prophylaxis were doxycycline (n = 282, 48%), atovaquone/proguanil (n = 205, 34.7%), and mefloquine (n = 57; 9.6%). The other prescriptions were choice left to patients among the three preferred options (n = 16; 2.7%), chloroquine/proguanil (n = 27; 4.5%), and chloroquine (n = 4; 0.7%). Prescriptions were in accordance to the French 2008 recommendations in 95.1% (95% CI: 93–96.5%) of the cases (578 of 608). Non-conforming

prescriptions are detailed BIBW2992 manufacturer in Figure 1. Four patients were prescribed chloroquine/proguanil although the trip was planned to an area with high prevalence of resistance in sub-Saharan Africa: Senegal, Mali, and Burkina Faso (which changed from area 2 to area 3 during the study in the June 2008 issue of the BEH). One of these travelers came

back with Plasmodium falciparum malaria. He presented a mild case and was treated as an outpatient with atovaquone/proguanil with a favorable outcome. Another patient received a prescription of chemoprophylaxis without indication. He got a prescription of atovaquone/proguanil for a trip to Vietnam, but he visited only Hanoi where MG132 there is no risk for malaria. The reasons given by physicians for their choices of malaria prophylaxis were cost (n = 282; 49%), tolerability (n = 74; 13%), duration of prophylaxis after exposure (n = 160; 29%), benefit of a weekly regimen (n = 31; 5%) or because a similar prescription had already been

provided by the general practitioner (n = 73; 13%). All travelers to tropical areas should have been advised to use personal protection against insect-borne diseases, not only for malaria but also for dengue and other arthropod-borne diseases. In files, mosquito nets were recommended in 549 (77.7%) instances, insect repellents in 675 (93.4%), and impregnated clothings in 663 (92.1%). Among the group of patients returning to their country to visit friends and relatives (n = 120), mosquito nets were recommended in 81 cases (62%) and insecticides in 106 cases (92.4%). With regards to malaria-endemic areas (n = 608), repellents were prescribed for 538 travelers (93.4%), impregnated clothings for 560 (92.1%), and mosquito nets for 473 (77.7%). Among the 730 travelers, 542 (74.2%) went to yellow fever-endemic areas. Yellow fever vaccine was contra-indicated in three immunocompromised patients who were not vaccinated.

Surface-sterilized wheat seeds were treated with bacterial cultur

Surface-sterilized wheat seeds were treated with bacterial cultures at 108 CFU mL−1 as described previously (Pierson et al., 1998). To determine bacterial colonization, sterile soil and natural soil (not autoclaved) were used to grow wheat seeds. After 7, 14 and 21 days, 10 plants were collected HSP activation randomly from each treatment, and the population densities in the rhizosphere (1 g) and the root tip (1 cm) were determined as described by Hoben & Somasegaran (1982). The experiment was repeated twice, and population data collected as CFU counts were log10-transformed

before statistical analysis. Data were analyzed and compared by performing two-sample independent t-tests (P<0.05 was considered significant) using origin 7.0 software (Originlab Corporation). Strain 2P24 carrying a plasmid-borne phlA-lacZ fusion was subjected to a random mini-Tn5 insertion mutagenesis to identify novel regulators of the antibiotic 2,4-DAPG production. Among the ∼10 000 transposon mutants tested, one mutant designated PMphlA23, which exhibited the greatest reduction in phlA Selleck PXD101 expression (83% decrease of the β-galactosidase activity compared with its parental strain), was selected and purified for further studies. The mini-Tn5-flanking

region in the PMphlA23 mutant was cloned, and sequence analysis revealed that the transposon was inserted into the upstream region between positions −16 and −17 of a locus that had 84% amino acid identity to Hfq (Fig. 1), encoding a key global regulator for stress resistance and virulence in Pseudomonas aeruginosa (Sonnleitner et al., 2003). A 3.2-kb BamHI fragment containing the entire hfq gene was cloned from the genomic DNA of strain 2P24 (see Materials and methods). Sequencing and blast analysis (Altschul et al., 1997) of this fragment revealed three ORFs (Fig. 1). The deduced

product (86 amino acids) of the hfq gene in strain 2P24 is very similar to the Hfq proteins of P. fluorescens Pf0-1 (accession number CP000094; 98% identity), Pseudomonas syringae pv. tomato DC3000 (accession number AAO58370; 95% identity), P. aeruginosa PAO1 (accession number AE004091; 84% identity) and E. coli O157:H7 (accession number AAG59368; 60% identity). G protein-coupled receptor kinase As in P. aeruginosa PAO1 (Sonnleitner et al., 2003), the hfq gene in P. fluorescens 2P24 is localized between two ORFs, encoding a putative tRNA isopentenyltransferase (OrfA, 79% identity to gene PA4945 of P. aeruginosa PAO1) and a putative GTP-binding protein (OrfB, 83% identity to gene PA4943 of P. aeruginosa PAO1). This arrangement also appears to be conserved in other Pseudomonas spp. (data not shown). In order to determine the potential regulatory effect of the hfq gene on phlA expression, the p970Gm-phlA plasmid containing the phlA promoter fused to the lacZ (β-galactosidase) reporter gene was transformed into the hfq mutant PM107 and its parental strain 2P24.

These results suggest that modulation of PI3K signaling could con

These results suggest that modulation of PI3K signaling could contribute to improve the cognitive deficits associated with FXS. “
“In the anti-saccade task, a subject must make a saccadic eye movement in the opposite direction from a suddenly-presented visual target. This sets up a conflict between the natural tendency to make a pro-saccade towards the target and the required anti-saccade. Consequently there is a tendency to make errors, usually corrected by a second movement in the

correct anti-saccade direction. In a previous paper, we showed that a very simple model, with racing LATER (Linear Approach to Threshold at Ergodic Rate) units for the pro- and anti-directions, and a stop unit that inhibits the impending error response, could account precisely for the detailed distributions of reaction times both for correct and ABT-199 clinical trial error responses. However, the occurrence and timing of these final corrections have not been studied. We propose a novel mechanism: the decision race re-starts Selumetinib order after an error. Here we describe measurements of all the responses in an anti-saccade task, including corrections,

in a group of human volunteers, and show that the timing of the corrections themselves can be predicted by the same model with one additional assumption, that initiation of an incorrect pro-saccade also resets and initiates a corrective anti-saccade. No extra parameters are needed to predict this complex aspect of behaviour, adding weight to our proposal that we correct our mistakes by re-starting a neural decision race.

The concept of re-starting Liothyronine Sodium a decision race is potentially exciting because it implies that neural processing of one decision can influence the next, and may be a fruitful way of understanding the complex behaviour underlying sequential decisions. “
“Axonal projections in the CNS can be categorized as either crossed or uncrossed. Crossing and uncrossing of axons has been explained by attractive and repulsive molecules like Netrin-1 and Slits, which are secreted by midline structures. However, uncrossed projections can be established even in double knockout mice of slit1 and slit2 or of roundabout1 (robo1) and robo2, two receptors for Slits. Here, we found that a novel mechanism mediated by Neuropilin-2 (Nrp2) contributes to the formation of uncrossed projections of midbrain dopaminergic neurons (mDANs). Nrp2 transcriptional activities were detected in a subset of mDANs, and its protein was expressed in mDAN axons growing through the ipsilateral diencephalon. In nrp2lacZ/lacZ mice, mDAN axons aberrantly grew toward the ventral midline and even crossed it, suggesting that Nrp2 is necessary for the development of mDAN ipsilateral projections. We investigated the involvement of Semaphorin 3B (Sema3B) and Sema3F, two ligands of Nrp2, by analysing mDAN axon trajectories in single or double knockout mice.

Similarly, bacteria express a variety of regulatory RNA species r

Similarly, bacteria express a variety of regulatory RNA species ranging from trans-acting RNAs (sRNA), cis-acting RNAs (riboswitches), antisense RNAs and protein-interacting RNAs (6S RNA, CsrB-like RNAs) (Waters & Storz, 2009), and while our knowledge on these species is currently mostly based on E. coli, this is likely to change with the advent of sequencing-based transcriptomics. When combined with buy GW-572016 the latest developments in microarray technologies,

like high-density tiling microarrays (Rasmussen et al., 2009; Toledo-Arana et al., 2009), we now have the ability to investigate transcription at single-nucleotide resolution. This is likely to enrich our knowledge of microbial diversity, and will undoubtedly show us the many different approaches used by bacteria to solve the problems encountered in their respective niches. The author thanks the members of the research group and the collaborators, as well as three anonymous reviewers for helpful comments and suggestions. Research at the author’s laboratory is supported by the BBSRC Institute Strategic Programme Grant to the IFR. “
“Carotenoids

are a structurally diverse class of terpenoid pigments that are synthesized by many microorganisms and plants. In this study, we identified five putative carotenoid biosynthetic PLX4032 concentration genes from the ascomycete Gibberella zeae (GzCarB, GzCarO, GzCarRA, GzCarT, and GzCarX). HPLC showed that the fungus produces two carotenoids: neurosporaxanthin and torulene. We deleted

the five genes individually to determine their functions. GzCarB, GzCarRA, and GzCarT were required for neurosporaxanthin biosynthesis, but the deletion of GzCarX or GzCarO (ΔgzcarX or ΔgzcarO) failed to alter the production of neurosporaxanthin mafosfamide or torulene. ΔgzcarRA and ΔgzcarB did not produce neurosporaxanthin or torulene. ΔgzcarB led to the accumulation of phytoene, which is an intermediate in carotenoid biosynthesis, but ΔgzcarRA did not. ΔgzcarT produced torulene but not neurosporaxanthin. Based on these functional studies and similarities to carotenoid biosynthesis genes in other fungi, we deduced the functions of the three genes and propose the carotenoid biosynthetic pathway of G. zeae. Carotenoids are important natural terpenoid pigments produced by many microorganisms and plants, but not animals. Of the >700 natural carotenoids that have been identified, most are C40 terpenoids that vary in the number of conjugated double bonds, end-group structures, and oxygen-containing functional groups (Britton et al., 2004). The interesting properties and human health benefits of carotenoids have received much attention. Carotenoids exhibit significant anticarcinogenic and antioxidant activity, and play an important role in preventing chronic disease (Landrum & Bone, 2001). Carotenoids are derived from the isoprenoid biosynthetic pathway (Umeno et al., 2005).