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Hepatocytes constitute the majority of hepatic cells, and play a key role in controlling systemic innate immunity, via pattern-recognition receptors (PRRs) and by synthesizing complement and acute phase proteins. Leishmania infantum, a protozoan parasite that causes human and canine leishmaniasis, infects liver by establishing inside the Kupffer cells. The current study proposes the elucidation of the immune response generated by dog hepatocytes when exposed to L. infantum. Additionally, the impact of adding leishmanicidal compound, meglumine antimoniate (MgA), to parasite-exposed hepatocytes was also addressed. L. infantum presents a high tropism to hepatocytes, establishing strong membrane interactions. The possibility of L. infantum internalization by hepatocytes was raised, but not confirmed. Hepatocytes were able to recognize parasite presence, inducing PRRs [nucleotide oligomerization domain (NOD)1, NOD2 and Toll-like receptor (TLR)2] gene expression and generating a mix pro- and anti-inflammatory cytokine response. Reduction of cytochrome P 450s enzyme activity was also observed concomitant with the inflammatory response. Addition of MgA increased NOD2, TLR4 and interleukin 10 gene expression, indicating an immunomodulatory role for MgA. Hepatocytes seem to have a major role in coordinating liver's innate immune response against L. infantum infection, activating inflammatory mechanisms, but always balancing the inflammatory response in order to avoid cell damage.
The use of cactus cladodes in animal feed is well-established in semi-arid areas. The cactus Nopalea cochenillifera (L.) Salm-Dyck cladodes (Nopalea) have high acceptability amongst dairy cows and are resistant to carmine cochineal insects (Dactylopius opuntiae Cockerell), a problem in semi-arid regions, but in regions of prolonged drought, it has lower productivity compared with the cactus Opuntia stricta (Haw.) Haw cladodes (Opuntia), which is also resistant to the insect. The objective of the current study was to evaluate the intake and content of digestible material of dry matter (DM) and its components, feeding behaviour, microbial protein synthesis, nitrogen balance, blood parameters, performance and milk composition of Holstein cows fed a control diet, containing either Nopalea or Opuntia associated with different concentrate levels (225, 275, 325 and 375 g/kg). Ten cows with an initial average milk production of 20 ± 2.1 kg/day were distributed into a double 5 × 5 Latin square design. Diets containing 775 g roughage/kg and 225 g concentrate/kg promoted similar responses to the analysed variables regardless of the cactus cladode used, except for digestibility of neutral detergent fibre. Diets containing higher proportions of concentrate (325 and 375 g/kg) promoted greater DM intake and 3.5% fat-corrected milk yield. The diet containing Opuntia at 775:225 g/kg roughage:concentrate proportion is as effective as the control diet for Holstein cows producing 20 kg of milk/day. To promote greater milk production, higher proportions of concentrate should be added to diets using Opuntia.
Foetal sex hormones can have powerful and far-reaching effects on later phenotype. However, obtaining accurate measurements is difficult for ethical reasons, and researchers often employ proxy variables to examine their effects. The relative length of the second and fourth fingers (digit ratio or 2D:4D) is frequently used for this purpose, as it is hypothesized to index variance in prenatal androgen and oestrogen exposure. Most studies employing this method examine digit ratio for the right hand (R2D:4D) and/or left hand (L2D:4D), though the mean value (M2D:4D) (i.e., the average of R2D:4D and L2D:4D) and directional asymmetry (D[R–L]) (i.e., R2D:4D minus L2D:4D) are also commonly used. As no published studies have examined M2D:4D or D[R-L] in relation to testosterone measured from amniotic fluid, we conducted a secondary analysis of data published by Ventura et al. The sample comprises 106 mothers from Portugal who underwent amniocentesis during the second trimester and their neonates. Newborn M2D:4D was negatively correlated with amniotic testosterone in females (P<0.05) but not in males; no significant association was observed between amniotic testosterone and D[R–L] in either sex. In addition, we examined testosterone measured from maternal circulation during the second trimester, and found that it was not a significant predictor of M2D:4D or D[R–L] in male or female infants. Further research should aim to measure the ratio of testosterone to oestradiol present in amniotic fluid and maternal plasma, to examine whether either is a predictor of digit ratio variables at different stages of postnatal development.
This study investigates how consistent genetic factors are, as measured by heritability estimates (h2), in the leisure-time physical activity index (LTPAI) and sport participation index (SPI) from early (10–14 yrs) to late adolescence (15–19 yrs). The sample comprises 12,385 subjects from 3,378 Portuguese nuclear families. Height and weight were measured and body mass index (BMI) was calculated, and the LTPAI and SPI were estimated by questionnaire. Socioeconomic status (SES) was assessed by parental occupation. Analyses were done using S.A.G.E. software. Our results showed that h2 estimates for the LTPAI and SPI in the two age groups (10–14 yrs and 15–19 yrs) were stable: for the LTPAI, h2 = 0.297 and 0.322, respectively; and for the SPI, h2 = 0.413 and 0.428, respectively. Sibling correlations and environmental correlations are higher in the younger age group for both the LTPAI and the SPI. Spousal correlations are higher in the younger age group for the LTPAI and lower for the SPI than the older group. Parent–offspring correlations are similar in both age groups for the LTPAI and SPI. In conclusion, the influence of genetic factors on physical activity and sport participation remains stable across age in adolescence. However, variation in sibling correlations — in particular, environmental correlations — was observed. These findings suggest that shared/non-shared environmental factors express different degrees of importance across age. Future intervention programs aiming to promote change in behaviors need to consider these results to bring about positive changes in physical activity and sport participation behaviors within the family setting.
In this study, a Leishmania hypothetical protein, LiHyS, was evaluated regarding its antigenicity, immunogenicity and protective efficacy against visceral leishmaniasis (VL). Regarding antigenicity, immunoblottings and an enzyme-linked immunosorbent assay using human and canine sera showed high sensitivity and specificity values for the recombinant protein (rLiHyS) in the diagnosis of VL. When evaluating the immunogenicity of LiHyS, which is possibly located in the parasite's flagellar pocket, proliferative assays using peripheral blood mononuclear cells from healthy subjects or VL patients showed a high proliferative index in both individuals, when compared to the results obtained using rA2 or unstimulated cultures. Later, rLiHyS/saponin was inoculated in BALB/c mice, which were then challenged with Leishmania infantum promastigotes. The vaccine induced an interferon-γ, interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor production, which was maintained after infection and which was associated with high nitrite and IgG2a antibody levels, as well as low IL-4 and IL-10 production. Significant reductions in the parasite load in liver, spleen, bone marrow and draining lymph nodes were found in these animals. In this context, the present study shows that the rLiHyS has the capacity to be evaluated as a diagnostic marker or vaccine candidate against VL.
We studied the ontogenetic growth of goat wethers (castrated male goats) of the Saanen and Swiss Alpine breeds based on a large range of intraspecific body mass (BM). The body parts and the chemical constituents of the empty body were described by the allometric function by using BM and the empty body mass (EBM) as the predictors for morphological traits and chemical composition, respectively. We fitted the allometric scaling function by applying the SAS NLMIXED procedure, but to evaluate assumptions regarding variances in morphological and compositional traits, we combined the scaling function with homoscedastic (MOD1), and the heteroscedastic exponential (MOD2) and power-of-the-mean (MOD3) variance functions. We also predicted the ontogenetic growth by using the traditional log-log transformation and back-transformed results into the arithmetic scale (MOD4). We obtained predictions from MOD4 in the arithmetic scale by a two-step process, and evaluated MOD1, MOD2 and MOD3 by a model selection framework, and compared MOD4 with MOD1, MOD2 and MOD3 based on goodness-of-fit measures. Based on information criteria for model selection, heterogeneous variance functions were more likely to describe 10 over 36 traits with a low level of model selection uncertainty. One trait was predicted by averaging the MOD1 and MOD2 variance functions; and nine traits were better described by averaging the MOD2 and MOD3 variance functions. The predictions for other 16 traits were averaged from MOD1, MOD2 and MOD3. However, MOD4 better described 11 traits according to the goodness-of-fit measures. Depending on the variable being analyzed, the body parts and the chemical amounts exhibited the three types of allometric behavior with respect to BM and EBM, that is, positive, negative and isometric ontogenetic growth. Reference BMs, that is, 20, 27, 35 and 45 kg, were used to compute the net protein and energy requirements based on the first derivative of the scaling function, and the results were presented in reference to the EBM and EBM0.75. Both the net protein and energy requirements scaled to EBM0.75 increased from 20 to 45 kg of BM.
Objectives: This study examined whether children with distinct brain disorders show different profiles of strengths and weaknesses in executive functions, and differ from children without brain disorder. Methods: Participants were children with traumatic brain injury (N=82; 8–13 years of age), arterial ischemic stroke (N=36; 6–16 years of age), and brain tumor (N=74; 9–18 years of age), each with a corresponding matched comparison group consisting of children with orthopedic injury (N=61), asthma (N=15), and classmates without medical illness (N=68), respectively. Shifting, inhibition, and working memory were assessed, respectively, using three Test of Everyday Attention: Children’s Version (TEA-Ch) subtests: Creature Counting, Walk-Don’t-Walk, and Code Transmission. Comparison groups did not differ in TEA-Ch performance and were merged into a single control group. Profile analysis was used to examine group differences in TEA-Ch subtest scaled scores after controlling for maternal education and age. Results: As a whole, children with brain disorder performed more poorly than controls on measures of executive function. Relative to controls, the three brain injury groups showed significantly different profiles of executive functions. Importantly, post hoc tests revealed that performance on TEA-Ch subtests differed among the brain disorder groups. Conclusions: Results suggest that different childhood brain disorders result in distinct patterns of executive function deficits that differ from children without brain disorder. Implications for clinical practice and future research are discussed. (JINS, 2017, 23, 529–538)
Introduction: Increased prescribing of high potency opioids has been associated with increasing opioid addiction and linked to serious adverse outcomes including misuse, diversion, overdose and death. Problems related to opioids are a major Canadian public health concern yet few data are available on prescribing in most Canadian provinces. The objective of this study was to describe opioid prescribing in Ontario EDs and patient harms associated with this practice. Methods: We conducted a population-based cohort study among Ontario residents aged 15-64 years who were eligible for public drug coverage between April 2008 and March 2012. Using administrative databases, we identified patients with no opioid use in the past 12 months who received a prescription opioid from an emergency or family physician. Patients were followed for 2 years following their index prescription. The primary outcome was hospital admission for opioid toxicity and secondary outcome was dose-escalation exceeding 200 mg morphine equivalents (MEQ). Results: Of the 77,270 unique patients included, 33,492 (43.3%) and 43,778 (56.7%) prescriptions were issued by emergency physician (EP) and family physicians (FP), respectively. FP patients were older (45.9 vs 41.2 yr, MSD 0.35), had fewer ED visits (0.9 vs 2.3, MSD 0.46), and more FP visits (11.5 vs 8.7 MSD 0.31) in the year prior to their index visit. For combination products, EPs were more likely to prescribe oxycodone compared to FPs (37.2% vs 16.7%, Δ 20.5, 95% CI: 19.9, 21.2). For single agent products, EPs were more likely to prescribe hydromorphone compared to FPs (44.5% vs 21.7%, Δ 22.8, 95% CI: 20.4, 25.2). FPs were more likely to prescribe codeine either as a combination or single agent formulation. EP prescriptions led to significantly more hospital admissions for opioid toxicity (0.5% vs 0.3%, Δ 0.2, 95% CI: 0.1, 0.3), while FP prescriptions more often resulted in dose escalation beyond 200 mg MEQs (0.1% vs 0.7%, Δ 0.6, 95% CI: 0.4, 0.7). Conclusion: A large percentage of opioid-naïve patients receive an initial opiate prescription in the ED, where the use of high potency opioids is much more common, with 1/200 of these patients subsequently hospitalized for opioid toxicity. Creation of a physician accessible provincial registry would be useful to monitor opioid prescribing and dispensing, inform clinical practice, and identify patients at high-risk who may benefit from early interventions.
Zika virus infection was declared a public health emergency of international concern in February 2016 in response to the outbreak in Brazil and its suspected link with congenital anomalies. In this study, we use notification data and disease natural history parameters to estimate the basic reproduction number (R0) of Zika in Rio de Janeiro, Brazil. We also obtain estimates of R0 of dengue from time series of dengue cases in the outbreaks registered in 2002 and 2012 in the city, when DENV-3 and DENV-4 serotypes, respectively, had just emerged. Our estimates of the basic reproduction number for Zika in Rio de Janeiro based on surveillance notifications (R0 = 2·33, 95% CI: 1·97–2·97) were higher than those obtained for dengue in the city (year 2002: R0 = 1·70 [1·50–2·02]; year 2012: R0 = 1·25 [1·18–1·36]). Given the role of Aedes aegypti as vector of both the Zika and dengue viruses, we also derive R0 of Zika as a function of both dengue reproduction number and entomological and epidemiological parameters for dengue and Zika. Using the dengue outbreaks from previous years allowed us to estimate the potential R0 of Zika. Our estimates were closely in agreement with our first Zika's R0 estimation from notification data. Hence, these results validate deriving the potential risk of Zika transmission in areas with recurring dengue outbreaks. Whether transmission routes other than vector-based can sustain a Zika epidemic still deserves attention, but our results suggest that the Zika outbreak in Rio de Janeiro emerged due to population susceptibility and ubiquitous presence of Ae. aegypti.
We discus here the prehistoric settlement of the central-south Brazilian coast, and, more specifically, 1 old radiocarbon date obtained for a costal shellmound, as well as its implications concerning the chronology attributed to the settlement process. The accelerator mass spectrometry (AMS) technique was used to determine the 14C age of charcoal from a shellmound on the southern coast of Rio de Janeiro. The resulting age was 7860 ± 80 BP, an unexpected result that reinforces 2 similar previously obtained dates for the same region. Brazilian archaeologists, however, have questioned those 2 dates, because they would predate by some 2000 yr the antiquity consensually accepted for the settlement of the central-south Brazilian littoral.
Kawasaki disease is an acute systemic vasculitis. Cardiac complications are frequent and include endothelial dysfunction in patients with coronary anomalies. Thus far, endothelial dysfunction in patients with no coronary lesions is poorly understood. Our aim was to access the vascular function in adolescents and young adults long term after Kawasaki disease, but without coronary aneurysms or any other cardiac risk factors.
We carried out a single-centre prospective study in a Portuguese population. We evaluated two groups of subjects: (1) Kawasaki disease patients over 11 years of age, diagnosed >5 years ago, with no coronary lesions or any other risk factors for cardiovascular disease; (2) control group of individuals without cardiovascular risk factors. Patients and controls were clinically assessed. Endo-PAT and carotid intima-media thickness assessment were performed to determine vascular function.
In total, 43 Kawasaki disease patients were assessed and compared with 43 controls. Kawasaki disease patients presented a decreased reactive hyperaemia index compared with controls (1.59±0.45 versus 1.98±0.41; p<0.001). Augmentation index was similar in both groups (−4.5±7 versus −5±9%; p 0.6). The mean carotid intima-media thickness was not significantly increased in the Kawasaki disease group. There were no statistically significant changes with regard to laboratory data.
Children with Kawasaki disease may have long-term sequelae, even when there is no discernible coronary artery involvement in the acute stage of the disease. Further research is needed to assess whether known strategies to improve endothelial function would bring potential benefits to Kawasaki disease patients.
After 22 yr of the low-level liquid scintillation counting 14C laboratory at the Center for Nuclear Energy in Agriculture (CENA) at São Paulo University (USP), Piracicaba, Brazil, and several collaborative projects with Brazilian and international researchers from distinct scientific areas, the first 14C accelerator mass spectrometry (AMS) laboratory in Latin America was installed at the Physics Institute of the Universidade Federal Fluminense (UFF), Niterói, Brazil. A 250kV single stage accelerator produced by National Electrostatics Corporation began its operation in 2012. In this work, we compare measurements performed at the AMS Radiocarbon Laboratory at UFF (LAC-UFF) with those performed at CENA and the University of Georgia (UGAMS), Georgia, USA. All the results obtained from distinct inorganic and organic samples were in very good agreement.
Frangula azorica V. Grubow is a Macaronesian flora medicinal plant, endemic from Azores islands and inscribed on the International Union for the Conservation of Nature (IUCN) Red List of Threatened Species. This species, known as “sanguinho,” belongs to the family Rhamnaceae, the same as Frangula alnus Mill. and Frangula purshiana (DC.) J. G. Cooper, two widely used official laxative herbal medicines of the western Pharmacopoeias constituted by the dried barks of each species. Morphological and chemical studies on F. azorica dried bark are scarce although it is potentially recognized as a Portuguese laxative herbal medicine. Macroscopically, the bark occurs in quills or nearly flat pieces. A channeled external surface with transversely elongated lenticels is characteristic. When the outer phellem layer is removed, a bright purple inner phellem layer is disclosed. Light and electron microscopy observations revealed flattened phellem cells with slightly thickened walls, cortical parenchyma with secretory ducts and groups of sclereids, phloem with groups of fibers and sheaths of parenchymatous cells containing druses or more frequently prismatic crystals of calcium oxalate, and parenchymatous medullary rays one to three cells wide with spherical starch grains. Observation of these botanical characteristics must be included in quality monographs of F. azorica bark herbal medicine.
Rickettsia rickettsii infection is being increasingly recognized as an important cause of fatal acute illness in Brazil, where this tick-borne disease is designated Brazilian spotted fever (BSF). In this study we report five fatal cases of BSF in employees of an animal shelter in an urban area in the municipality of Rio de Janeiro in southeast Brazil after a natural disaster on 11 January 2011. Four of the cases occurred from 27 January to 11 April 2011, while the fifth fatal case was identified in April 2012. Three cases were confirmed by molecular analysis and two by epidemiological linkage. An investigation of BSF was performed in the animal shelter, and blood samples were collected from 115 employees and 117 randomly selected dogs. The presence of high levels (1024–4096) of antibodies against spotted fever group rickettsiae was found in three (2·6%) employees and 114 (97·5%) dogs. These findings emphasize the need to consider BSF as a possible cause of undifferentiated febrile illness, especially dengue and leptospirosis, in patients occupationally exposed to dogs heavily infested by ticks, mainly working at kennels and animal shelters that have inadequate space for the animals housed and frequently providing an environment conducive to exposure to pathogens such as R. rickettsii.
Many dog owners see homemade diets as a way of increasing the bond with their pets, even though they may not have the convenience of commercial diets. Modifications of ingredients, quality and proportion might change the nutritional composition of the diet, generating nutritional imbalances. The present study evaluated how dog owners use and adhere to homemade diets prescribed by veterinary nutritionists over an extended period of time. Forty-six owners of dogs fed a homemade diet for at least 6 months were selected for the present study. Owners were invited to answer questions by first reading all possible answers and then selecting the one that best indicated their opinion. The results were evaluated through descriptive statistics. Thirty-five owners (76·1 %) found that the diets are easy to prepare. Fourteen owners (30·4 %) admitted to modifying the diets, 40 % did not adequately control the amount of provided ingredients, 73·9 % did not use the recommended amounts of soyabean oil and salt, and 34·8 % did not correctly use the vitamin, mineral or amino acid supplements. Twenty-six owners (56·5 %) reported that their dogs refused to eat at least one food item. All of these alterations make the nutritional composition of the diets unpredictable and likely nutritionally imbalanced. Although homemade diets could be a useful tool for the nutritional management of dogs with certain diseases, not all owners are able to appropriately use this type of diet and adhere to it for an extended period of time and this limitation needs to be considered when recommending the use of homemade diets.
In this work, we present an analysis of SNe number ratios in spiral galaxies with different morphological subtypes, luminosities, sSFR, and metallicities, to provide important information about the physical properties of the progenitor populations.
Stray cats live in high-density colonies in urban areas and pose a health hazard to household cats and humans. In Portugal, information on the parasitic fauna of stray cats is limited and relies mostly on results from faecal analysis. The present survey aimed to determine the prevalence, diversity and intensity of parasites in stray cats from the urban area of Lisbon by means of parasitological necropsy. Internal organs were collected from 162 cats captured in different areas of the city and systematically subjected to parasitological dissection. Helminths were identified by macro- and microscopic examination and protozoa by faecal floatation and sedimentation techniques. The overall prevalence of parasites was 90.7% (95% confidence interval (CI): 85.3–94.6%). A total of 12 parasite species was recorded: Cystoisospora felis (14.2%), Cystoisospora rivolta (46.3%), Sarcocystis sp. (1.2%), Ancylostoma tubaeforme (19.1%), Toxocara cati (38.3%), Ollulanus tricuspis (30.9%), Aelurostrongylus abstrusus (12.4%), Eucoleus aerophilus (0.6%), Taenia taeniaeformis (3.1%), Dipylidium caninum (53.1%), Joyeuxiella pasqualei (15.4%) and Diplopylidium nölleri (3.7%). Overall mean species richness was 2.36 ± 1.52. Helminth mean intensity was highest for O. tricuspis (285.8), followed by D. caninum (42.4), J. pasqualei (14.4), A. tubaeforme (8.1) and T. cati (5.9). The prevalence and variety of parasites found in our sampling are substantially higher than the numbers previously reported in Portugal. Some of the parasites, including T. cati and A. tubaeforme, are zoonotic, which emphasizes the need for parasite control strategies based on demographic containment of stray cat populations in urban areas to promote public health protection.
We observe the heat flux exchanged by the hot tip of a scanning thermal microscope, which is an instrument based on the atomic force microscope. We first vary the pressure in order to analyze the impact on the hot tip temperature. Then the distance between the tip and a cold sample is varied down to few nanometers, in order to reach the ballistic regime. We observe the cooling of the tip due to the tip-sample heat flux and compare it to the current models in the literature.
SMYB1 is a Schistosoma mansoni protein highly similar to members of the Y-box binding protein family. Similar to other homologues, SMYB1 is able to bind double- and single-stranded DNA, as well as RNA molecules. The characterization of proteins involved in the regulation of gene expression in S. mansoni is of great importance for the understanding of molecular events that control morphological and physiological changes in this parasite. Here we demonstrate that SMYB1 is located in the cytoplasm of cells from different life-cycle stages of S. mansoni, suggesting that this protein is probably acting in mRNA metabolism in the cytoplasm and corroborating previous findings from our group that showed its ability to bind RNA. Protein–protein interactions are important events in all biological processes, since most proteins execute their functions through large supramolecular structures. Yeast two-hybrid screenings using SMYB1 as bait identified a partner in S. mansoni similar to the SmD3 protein of Drosophila melanogaster (SmRNP), which is important in the assembly of small nuclear ribonucleoprotein complexes. Also, pull-down assays were conducted using immobilized GST-SMYB1 proteins and confirmed the SMYB1-SmRNP interaction. The interaction of SMYB1 with a protein involved in mRNA processing suggests that it may act in processes such as turnover, transport and stabilization of RNA molecules.
Both inhibitory-based executive functioning (IB-EF) and basic information processing (BIP) deficits are found in clinic-referred attention deficit hyperactivity disorder (ADHD) samples. However, it remains to be determined whether: (1) such deficits occur in non-referred samples of ADHD; (2) they are specific to ADHD; (3) the co-morbidity between ADHD and oppositional defiant disorder/conduct disorder (ODD/CD) has additive or interactive effects; and (4) IB-EF deficits are primary in ADHD or are due to BIP deficits.
We assessed 704 subjects (age 6–12 years) from a non-referred sample using the Development and Well-Being Assessment (DAWBA) and classified them into five groups: typical developing controls (TDC; n = 378), Fear disorders (n = 90), Distress disorders (n = 57), ADHD (n = 100), ODD/CD (n = 40) and ADHD+ODD/CD (n = 39). We evaluated neurocognitive performance with a Two-Choice Reaction Time Task (2C-RT), a Conflict Control Task (CCT) and a Go/No-Go (GNG) task. We used a diffusion model (DM) to decompose BIP into processing efficiency, speed–accuracy trade-off and encoding/motor function along with variability parameters.
Poorer processing efficiency was found to be specific to ADHD. Faster encoding/motor function differentiated ADHD from TDC and from fear/distress whereas a more cautious (not impulsive) response style differentiated ADHD from both TDC and ODD/CD. The co-morbidity between ADHD and ODD/CD reflected only additive effects. All ADHD-related IB-EF classical effects were fully moderated by deficits in BIP.
Our findings challenge the IB-EF hypothesis for ADHD and underscore the importance of processing efficiency as the key specific mechanism for ADHD pathophysiology.