The development of protection in mice was investigated for the in

The development of protection in mice was investigated for the intra-peritoneal and intranasal immunisation routes. The results showed that CpG ODN alone, or in combination with aluminum hydroxide, gave enhancement in anti-pertussis toxin, anti-filamentous haemagglutinin and especially

anti-pertactin titers after mucosal immunisation. Higher macrophage NO levels indicating activation were found when the antigens were co-formulated with CpG ODN. Vaccines containing CpG ODN gave enhanced humoral and CMI responses with a shift selleck toward Th-1 and increased protection against challenge infection with B. pertussis in mice.”
“Using a recently developed high-resolution learn more noninvasive ultrasonic method, we recently demonstrated that the intima-media complex of the common carotid artery show a bidirectional multiphasic longitudinal displacement of the same magnitude as the diameter change during the cardiac cycle. The longitudinal movement of the adventitial region was smaller, thus, we identified shear strain and, thus, shear stress, within the arterial wall. The aim of this study was to evaluate the intra-observer variability of measurement of the longitudinal displacement of the intima-media complex and the

intramural shear strain of the common carotid artery in vivo using the new ultrasonic method. The evaluation was carried out by comparing two consecutive check details measurements on the common carotid artery of 20 healthy human subjects. According to the method of Bland Altman, we show that

the systematic and random differences for the different phases of movement are acceptable in comparison to the measured displacement and no significant differences between the two measurements could be detected (p > 0.05 for all measured parameters). The coefficient of variation (CV) for measurement of the different phases of movement was <= 16%, including short-term physiologic variations. The higher variability in the measurement of the intramural shear strain (CV = 24%) has several explanations, which are discussed. In conclusion, this study shows that the present first ultrasonic method for high-resolution measurement of the longitudinal movement of the arterial wall is reliable and satisfactory for the further research of the longitudinal movement of the arterial wall in vivo. Further studies on the longitudinal movement of the arterial wall are important for developing an improved understanding of the physiology and the pathophysiology of the cardiovascular system. (E-mail: [email protected]) (c) 2010 World Federation for Ultrasound in Medicine & Biology.

Of the 10 adults (aged

28-55 years), 1 had 100% facial re

Of the 10 adults (aged

28-55 years), 1 had 100% facial repigmentation in 3 months, 1 had 76% to 99% facial repigmentation in 5 to 9 months, and 2 had 26% to 50% repigmentation in 3 months. Twelve patients developed some facial repigmentation. S63845 No patients experienced atrophy, telangiectases, or lesion enlargement during treatment. Combination calcipotriene 0.005%-betamethasone dipropionate 0.064% ointment shows promise as a once-daily vitiligo therapy. Adult and pediatric facial vitiligo patients may see repigmentation as early as 2 months after initiation of therapy. Children may experience a better response, but larger studies are needed. Cutis. 2011;88:256-259.”
“Introduction: Homozygosity for interleukin-6 (IL-6) 174G_C promoter polymorphism has recently been associated with indices of metabolic syndrome; however, this problem has not been investigated in non-obese subjects. The aim of this study was to explore the relation between abdominal fat distribution and some inflammatory risk factors of atheromatosis and IL-6 174G_C gene polymorphism in non-obese healthy subjects.\n\nMaterial and methods: Relationships were investigated between anthropometric variables, i.e. weight, height, BMI, waist circumference (WC), waist-to-hip ratio (WHR), body fat distribution (DXA), serum CRP and IL-6, insulin

sensitivity/resistance indices, and IL-6 Selleck Cyclopamine 174G_C gene polymorphism, in healthy non-obese Polish subjects: 232 women (age 31.4 +/- 5.5 years) and 199 men (age 30.3 +/- 6.0 years).\n\nResults: The genetic study revealed that the CC genotype was observed in 15.56% of subjects, the CG genotype in 52.74%, and the GG genotype in 31.7%. IL-6 and CRP

concentration did not differ among the genotypes. There were also no differences regarding BMI and WHR. The only differences among genotypes, observed only in men, were those concerning total fat (CC had higher fat content than CG and GG); the difference being statistically significant between CC and GG (p < 0.05), and gynoidal fat deposit (CC had higher gynoidal fat deposit than CG and GG); the difference being statistically significant between Citarinostat ic50 CC and GG (p < 0.025) and between CC and CG (p < 0.05). Biochemical parameters and insulin sensitivity did not differ among the genotypes.\n\nConclusions: These data show that IL-6 174G_C polymorphism is not associated with features describing metabolic syndrome in non-obese healthy subjects. (Pol J Endocrinol 2009; 60 (3): 172-179)”
“Objective. To evaluate the prevalence of atypical antipsychotic use in privately insured children and the diagnoses associated with treatment. Study design. Claims were used to conduct a retrospective cohort study of children aged 2 through 18 years in the Midwest, covered by private insurance between 2002 and 2005 (n = 172 766). The 1-year prevalence of children receiving atypical antipsychotics was determined along with associated diagnoses. Results.

The first arriving male is assured of access to a virgin female,

The first arriving male is assured of access to a virgin female, as his intermediate location between selleck inhibitor the vulva and subsequently arriving males blocks these competitors. Males deposit an adhesive gelatinous copulatory plug into and over the vulva before departing the female.

Fecundity was greater in larger mating clusters, but this was a function of a greater rate of molting which is a prerequisite for mating. Males compete for virgin females by emerging and molting to the adult stage earlier than females. Mating aggregations have previously only been examined in snakes, but these studies have tended to be observational as snakes offer a challenging system for study. The relatively easy to culture and manipulate mermithid system may offer a model for experimental studies of male-male competition, protandry, copulatory plugs and female choice in mating clusters. (C) 2014 Elsevier Inc. All rights reserved.”
“Twin-to-twin transfusion syndrome (TTTS) is a serious complication in about 10% to 20% of monozygous twin gestations with an incidence of 4% to 35% in the United States. Severe TTTS is reported to occur in 5.5% to 17.5% of cases.

TTTS is a progressive disease in which sudden deteriorations in clinical status can occur, leading to death of a co-twin. Up to 30% of survivors may have abnormal neurodevelopment as a result of the combination of profound antenatal insult and the complications of severe prematurity. BGJ398 ic50 This article presents an overview of what is known about the pathophysiology and the diagnosis of TTTS, the role of echocardiography in TTTS, treatment options available for TTTS, complications of treatment for TTTS, and short- and long-term outcomes of TTTS.”
“Maize (Zea mays L.) has a lot of dietary

and industrial value around the globe and its yield potential is being extremely affected by abiotic stresses i.e., drought and soil salinity/sodicity etc. However, damaging effects FAK inhibitor of abiotic stresses can be ameliorated by the combined application of plant growth promoting rhizobacteria (PGPR) and mineral fertilizers. Therefore this field trial was designed to explore the effect of PGPR containing ACC deaminase (1-aminocyclopropane-1-carboxylate deaminase) alone and in combination with mineral fertilizers on yield and nutrient uptake of maize under drought in saline field. Two PGPR strains S-1 (Pseudomonas syringae) and S-2 (Pseudomonas fluorescens) were applied to the maize seeds in separate and along with full and half dose of recommended NPK fertilizers. Drought stress was imposed generally on whole experiment at tasseling by withholding irrigation up to similar to 50% field capacity (FC) level. The PGPR strains significantly improved the yield of maize when applied alone and further promising results were obtained when applied with mineral fertilizers.

The combustion properties and thermal stability of PLA/APP/TT23 a

The combustion properties and thermal stability of PLA/APP/TT23 and PLA/APP/TT4 composites were evaluated by UL-94 burning tests, limiting oxygen index (LOI), cone calorimeter tests and TGA, the chemical structure of char residues were analyzed by FTIR and XPS. It can be concluded that PLA with JQ-EZ-05 30 wt % of APP/TT4 (weight ratio 5 : 1) achieved the greatest flame retardancy. Moreover, the continuous and expansionary char layer observed from SEM images proved better char forming ability of TT4 than that of TT23. (c)

2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42086.”
“Twenty-two compounds of substituted benzoylguanidine derivatives were designed and synthesized as potent NHE1 inhibitors. Twelve compounds showed more potent NHE1 inhibitory activity than cariporide. The activities of compounds 7e, 7h and 7j (IC50 = 0.073

+/- 0.021, 0.084 +/- 0.012 and 0.068 +/- 0.021 nmol/L, respectively) were two orders of magnitude higher than that of cariporide (30.7 +/- 2.5 nmol/L). Myocardial cells in vitro screening showed 7j had highlighted protective effect on cardiomyocytes subjected to hypoxia/reoxygenation. Thus it is valuable for further investigation. (C) 2011 Elsevier Masson SAS. LY2157299 molecular weight All rights reserved.”
“Protein kinase C (PKC) has been widely implicated in positive and negative control of cell proliferation. We have recently shown that treatment of non-small cell lung cancer (NSCLC) cells with phorbol 12-myristate 13-acetate (PMA) during G(1) phase inhibits the progression into S phase, an effect mediated by PKC delta-induced up-regulation of the cell cycle inhibitor p21(Cip1). However, PMA treatment in asynchronously growing NSCLC cells leads to accumulation of cells in G(2)/M. Studies in post-G(1) phases revealed that PMA induced an irreversible G(2)/M cell cycle arrest in NSCLC cells and conferred morphological and biochemical features of senescence, including elevated

SA-beta-Gal activity and reduced telomerase activity. Remarkably, this effect see more was phase-specific, as it occurred only when PKC was activated in S, but not in G(1) phase. Mechanistic analysis revealed a crucial role for the classical PKC alpha isozyme as mediator of the G(2)/M arrest and senescence, as well as for inducing p21(Cip1) an obligatory event for conferring the senescence phenotype. In addition to the unappreciated role of PKC isozymes, and specifically PKC alpha, in senescence, our data introduce the paradigm that discrete PKCs trigger distinctive responses when activated in different phases of the cell cycle via a common mechanism that involves p21(Cip1) up-regulation.”
“Objectives Previous studies in HIV-infected populations have yielded conflicting results on the effect of antiretroviral therapy (ART) on cognition.

Lymphocyte DNA damage was independently correlated with only TAS

Lymphocyte DNA damage was independently correlated with only TAS. Lymphocyte DNA damage was increased in hypertensive patients. The major increase in lymphocyte DNA damage was observed in newly diagnosed hypertensive patients compared with hypertensive patients who already on drug therapy. Decreased TAS levels, which reflect to increased check details oxidative stress, may be the reason of increased

total lymphocyte DNA damage in South Indian hypertensive patients. Journal of Human Hypertension (2010) 24, 475-482; doi: 10.1038/jhh.2009.100; published online 7 January 2010″
“Mitochondria are responsible for the oxidative phosphorylation process. Accordingly, putatively adaptive changes in their genomic features have been variously associated with major eco-physiological shifts in animal evolution, including increased metabolic rates and heat adaptation. Antarctic pycnogonids offer an interesting system to test whether the selective pressure for heat production and increased aerobic metabolism may be driving genomic changes like: (a) unusual compositional biases at the nucleotide and amino acid level, possibly related to cold adaptation; (b) an accelerated rate of mutations/genomic rearrangements, possibly related to the mutagenic effects of oxygen intermediates.

The complete mitochondrial genome (mtDNA) of the Antarctic sea spider Ammothea carolinensis Leach, 1814 (Arthropoda: Pycnogonida), the type species for the genus Ammothea, has been determined and is here compared

to known genomes from Antarctic and temperate species. We describe a marked heterogeneity in base composition SHP099 Others inhibitor skewness parameters as well as a strong signature of purifying selection toward an increase in thymines at second codon positions, possibly associated with an increased stability of hydrophobic inter-membrane domains. We further observe a fairly high rate of genomic changes, including a possible hot spot of recombination at the level of tRNA-Q. Nevertheless, these features IPI-145 clinical trial do not seem to be restricted to the two Antarctic pycnogonids analyzed, as to suggest a causal relationship between cold adaptation and genomic changes, and are better interpreted as basal features shared by the entire group. The relevance of the newly determined sequence for the phylogeny of pycnogonids, including its base composition and genomic rearrangements, is further discussed.”
“The removal of recalcitrant chemicals in wastewater treatment systems is an increasingly relevant issue in industrialized countries. The elimination of persistent xenobiotics such as endocrine-disrupting chemicals (EDCs) emitted by municipal and industrial sewage treatment plants remains an unsolved challenge. The existing efficacious physico-chemical methods, such as advanced oxidation processes, are resource-intensive technologies. In this work, we investigated the possibility to remove phenolic EDCs [i.e., bisphenol A (BPA)] by means of a less energy and chemical consuming technology.

Copyright (C) 2010 S Karger AG, Basel”
“Artificial activato

Copyright (C) 2010 S. Karger AG, Basel”
“Artificial activators designed using transcription activator like effector (TALE) technology have broad utility, but previous studies suggest that these monomeric proteins often exhibit low activities. VE-821 Here we demonstrate that TALE activators can robustly function individually or in synergistic combinations to increase expression of endogenous human genes over wide dynamic ranges. These findings will encourage applications of TALE activators for research and therapy, and guide design of monomeric TALE-based fusion proteins.”
“Proline-rich

tyrosine kinase 2 (PYK2) is a cytoplasmic, nonreceptor tyrosine kinase implicated in multiple signaling pathways. It is a negative

regulator of osteogenesis and considered a viable drug target for osteoporosis treatment. The high-resolution structures of the human PYK2 kinase domain with different inhibitor complexes establish the conventional bilobal kinase 17-AAG inhibitor architecture and show the conformational variability of the DFG loop. The basis for the lack of selectivity for the classical kinase inhibitor, PF-431396, within the FAK family is explained by our structural analyses. Importantly, the novel DFG-out conformation with two diarylurea inhibitors (BIRB796, PF-4618433) reveals a distinct subclass of non-receptor tyrosine kinases identifiable by the gatekeeper Met-502 and the unique hinge loop conformation of Leu-504. This

is the first example of a leucine residue in the hinge loop that blocks the ATP binding site in the DFG-out conformation. Our structural, biophysical, and pharmacological studies suggest that the unique features of the DFG motif, including Leu-504 hinge-loop variability, can be exploited for the development of selective protein kinase inhibitors.”
“The purpose of this study was to investigate the beneficial effects of particulate ostrich eggshell grafting on the healing of experimentally induced skull defects. The clinical, radiological, histological, Prexasertib and histomorphometrical findings of this material were compared with the results of commercially available demineralized bone matrix (DBM). The study was conducted on 18 adult New Zealand rabbits. One defect served as a control and the remaining ones either were filled with different sized eggshell particles or DBM, in each animal. Clinical and radiological inspections and histologic investigations of the animals were done at the 1st, 3rd, and 6th months of postoperative period. Radiologically, minimal bone regeneration was observed at the empty, control defect sites. The most advanced bone regeneration was in the DBM grafted defects. The eggshell particle grafted defect sites displayed weak bone regeneration at earlier stages, at 1st and 3rd months after operation when compared with demineralized bone matrix.

In addition, the proposed method of syndrome classification could

In addition, the proposed method of syndrome classification could also avoid two disadvantages in previous methods: linear relation of the clinical data and mutually exclusive symptoms among different syndromes.

And it could help exploit the latent relation between syndromes and symptoms more effectively. Better performance of syndrome classification is able to be achieved according to the experimental results and the clinical experts.”
“Aim: Huanglian-Jie-Du decoction DMH1 datasheet (HLJDD) is an important multiherb remedy in TCM, which is recently demonstrated to be effective to treat ischemic stroke. Here, we aimed to investigate the pharmacological mechanisms of HLJDD in the treatment of ischemic stroke using systems biology approaches. Methods: Putative targets of HLJDD were predicted using MetaDrug. An interaction network of putative HLJDD targets and known therapeutic targets for the treatment of ischemic stroke was then constructed, and candidate HLJDD targets were identified by calculating

topological features, including ‘Degree’, ‘Node-betweenness’, ‘Closeness’, and ‘K-coreness’. The binding efficiencies of the candidate HLJDD targets with the corresponding compositive compounds were further validated by a molecular docking simulation. Results: A total of 809 putative targets were obtained for 168 compositive compounds AG-120 purchase in HLJDD. Additionally, 39 putative targets were common to all four herbs of HLJDD. Next, 49 major nodes were identified as candidate HLJDD targets due to their network topological AZD4547 mw importance. The enrichment analysis based on the Gene Ontology (GO) annotation system and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway demonstrated that candidate HLJDD targets were more frequently involved in G-protein-coupled receptor signaling pathways, neuroactive ligand-receptor interactions and gap junctions, which all played important roles in the progression of ischemic stroke. Finally,

the molecular docking simulation showed that 170 pairs of chemical components and candidate HLJDD targets had strong binding efficiencies. Conclusion: This study has developed for the first time a comprehensive systems approach integrating drug target prediction, network analysis and molecular docking simulation to reveal the relationships between the herbs contained in HLJDD and their putative targets and ischemic stroke-related pathways.”
“Percutaneous nephrolithotomy (PCNL) is generally considered a safe technique offering the highest stone-free rates after the first treatment as compared to the other minimal invasive lithotripsy techniques. Still, serious complications although rare should be expected following this percutaneous procedure.


“Background: Adherence to
4 tube


“Background: Adherence to tuberculosis (TB) treatment is troublesome, due to long therapy duration, quick therapeutic response which allows the patient to disregard about the rest of their treatment and the lack of motivation on behalf of the patient for improved. The objective of this study was to develop and validate a scoring system to predict the probability of lost Rabusertib clinical trial to follow-up outcome in TB patients as a way to identify patients suitable for directly observed treatments (DOT) and other interventions to improve adherence.\n\nMethods: Two prospective cohorts, were used to develop

and validate a logistic regression model. A scoring system was constructed, based on the coefficients of factors 123 associated with a lost to follow-up outcome.

The probability of lost to follow-up outcome associated with each score was calculated. Predictions in both cohorts were tested using receiver operating characteristic curves (ROC).\n\nResults: The best model to predict lost to follow-up outcome included the following characteristics: immigration GDC-0994 (1 point value), living alone (1 point) or in an institution (2 points), previous anti-TB treatment (2 points), poor patient understanding (2 points), intravenous drugs use (IDU) (4 points) or unknown IDU status (1 point). Scores of 0, 1, 2, 3, 4 and 5 points were associated with a lost to follow-up probability of 2,2% 5,4% 9,9%, 16,4%, 15%, and 28%, respectively. The ROC curve for the validation group demonstrated a good fit (AUC: 0,67 [95% CI; 0,65-0,70]).\n\nConclusion: This model has a good capacity to predict a lost to follow-up outcome. Its use could help TB Programs to determine which patients are good candidates for DOT and other strategies to improve TB treatment adherence.”
“Motivation: Metabolite identification from tandem mass spectra is an important problem in metabolomics, underpinning subsequent metabolic modelling and network analysis. Yet, currently this task requires matching the observed spectrum against a database of reference spectra originating from similar equipment and closely matching operating parameters, a condition that is rarely satisfied in public repositories.

https://www.selleckchem.com/products/ferrostatin-1-fer-1.html Furthermore, the computational support for identification of molecules not present in reference databases is lacking. Recent efforts in assembling large public mass spectral databases such as MassBank have opened the door for the development of a new genre of metabolite identification methods.\n\nResults: We introduce a novel framework for prediction of molecular characteristics and identification of metabolites from tandem mass spectra using machine learning with the support vector machine. Our approach is to first predict a large set of molecular properties of the unknown metabolite from salient tandem mass spectral signals, and in the second step to use the predicted properties for matching against large molecule databases, such as PubChem.

Their fasting serum insulin, fasting glucose, insulin-like growth

Their fasting serum insulin, fasting glucose, insulin-like growth factor-1 (IGF-1) concentrations, and the homeostasis assessment model for insulin resistance (HOMA-IR) and beta-cell function (HOMA%) were evaluated. The values of HOMA-IR in CUG SGA were significantly higher than that in NCUG SGA (P=0.002) and AGA children (P=0.036), respectively. Correlation analysis revealed that the concentrations of fasting serum insulin were positively correlated with IGF-1 (r=0.443,

P=0.001) and Delta height standard deviation score (SDS; r=0.500, P=0.002) in <= 6-year- old SGA children, but only with Delta weight SDS (r=0.496, P=0.030) in >6-year-old children. In conclusion, SGA children with CUG in height and a higher body mass index are prone to the development of insulin resistance. Higher levels of insulin were closely correlated with the postnatal height CUG in young SGA children and with the weight CUG in old children.”
“Hospital surfaces play SHP099 an selleck chemical important role in nosocomial infection (NI), in that the health-care environment contains a diverse population of microorganisms. Antibiotic resistance is the ability of a micro-organism to withstand the effects of an antibiotic, which is a specific type of drug resistance. Antibiotic resistance evolves naturally from a natural selection through random mutation, but could also be engineered by other selections. The research was performed

with laboratory method in Esfahan City and the PF-03084014 Neuronal Signaling inhibitor study as a whole comprised 194 strains obtained from hospital surfaces’ samples. These strains were randomly selected

from different wards of the hospital with sterile swab and NB medium. According to the results, Staphylococcus spp. (54.7%), Bacillus spp. (25%) and Enterobacteriaceae (10.7%) consist of isolated bacteria. The results of this study show high frequency of antibiotic resistant strain on hospital surfaces. Establishing systems for monitoring antimicrobial resistance in hospitals and the community, and linking these findings to resistance and disease surveillance data is fundamental to developing treatment guidelines accurately and to assessing the effectiveness of interventions appropriately.”
“In prescribing natural compounds, it is important matching scientific names of medicinal materials which I want to use and those which have been found those effects. This point is also important in Oriental medicine but isn’t kept because of differences in traditional sorting system and latest sorting system, external forms which are difficult to sort, and so on.. Baekbuja ((sic)) is a good example. In traditional 4 Korean medicine, Aconitum koreanum Raymond (AKR) has been used as a Baekbuja, but in traditional Chinese medicine, Typhonium gigantum Engl. (TGE) has been used as a Baekbuja. Added to this, Helianthus tuberosis Linne (HTL) is used as an imitational Baekbuja in distribution channels and prescriptions now.

Unlike canonical cadherins, it is believed to function primarily

Unlike canonical cadherins, it is believed to function primarily as a signaling molecule. T-cadherin is highly expressed in endothelium. Using transendothelial electrical resistance measurements and

siRNA-mediated depletion of T-cadherin in human umbilical vein endothelial cells, we examined its involvement in regulation of endothelial barrier. We found that in resting confluent monolayers adjusted either to 1% or 10% serum, T-cadherin depletion modestly, but consistently reduced transendothelial resistance. This was accompanied by increased phosphorylation of Akt and LIM kinase, reduced phosphorylation of p38 MAP kinase, but no 4EGI-1 difference in tubulin acetylation and in phosphorylation of an actin filament severing protein cofilin and myosin light chain kinase. Serum stimulation elicited a biphasic increase in resistance with peaks at 0.5 and 4-5 h, which was suppressed by a PI3 kinase/Akt inhibitor wortmannin and a p38 inhibitor SB 239063. T-cadherin depletion increased transendothelial resistance between the two peaks and reduced the amplitude of the second peak. T-cadherin depletion abrogated serum-induced Akt phosphorylation at Thr308 and reduced phosphorylation at Ser473, reduced phosphorylation of cofilin,

and accelerated tubulin deacetylation. selleck chemicals Adiponectin slightly improved transendothelial resistance irrespectively of T-cadherin depletion. T-cadherin depletion also resulted in a reduced sensitivity and delayed responses to thrombin. These data implicate T-cadherin in regulation of endothelial barrier function, and suggest a complex signaling network that links T-cadherin and regulation of barrier function. J. Cell. Physiol. 223: 94-102, 2010. (C) 2009 Wiley-Liss, Inc.”
“Advanced 432 image-guidance systems allowing presentation of three-dimensional navigational data in real time are being developed enthusiastically for many medical procedures. Other industries, including aviation and the military, have noted that shifting attention toward such compelling assistance

has detrimental effects. Using the detection rate of unexpected findings, we assess whether inattentional blindness is significant in a surgical context and evaluate the impact of on-screen navigational Nutlin-3 chemical structure cuing with augmented reality.\n\nSurgeons and trainees performed an endoscopic navigation exercise on a cadaveric specimen. The subjects were randomized to either a standard endoscopic view (control) or an AR view consisting of an endoscopic video fused with anatomic contours. Two unexpected findings were presented in close proximity to the target point: one critical complication and one foreign body (screw). Task completion time, accuracy, and recognition of findings were recorded.\n\nDetection of the complication was 0/15 in the AR group versus 7/17 in the control group (p = 0.008). Detection of the screw was 1/15 (AR) and 7/17 (control) (p = 0.041). Recognition of either finding was 12/17 for the control group and 1/15 for the AR group (p < 0.001).