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Background: The need to evaluate the impact of oral health has led to the development of instruments for measuring oral health-related quality of life (OHQoL). One such instrument is the Child Perceptions Questionnaire (CPQ11-14), developed specifically for 11-to-14-year-old children. As this questionnaire was considered long (37 items), shorter forms were developed with 8 (Impact Short Form: 8 - ISF:8) and 16 items (Impact Short Form: 16 - ISF:16) to facilitate use in the clinical setting and population-based health surveys. The aim of the present study was to translate and cross-culturally adapt these CPQ11-14 short forms for Brazilian Portuguese and evaluate the measurement properties of these versions for use on Brazilian children. Methods: Following translation and cross-cultural adaptation, the ISF:8 and ISF:16 were tested on 136 children from 11 to 14 years of age in the city of Belo Horizonte, Brazil. The instrument was administered by a trained researcher who also performed clinical examinations. The measurement properties (i.e. criterion validity, construct validity, internal consistency reliability, test-retest reliability) were determined. Discriminant validity was tested between groups, which were divided into children with no cavities and no malocclusion; children with cavities and without malocclusion; and children with malocclusion and without cavities. Results: The mean total score was 6.8 [standard deviation (SD) 4.2] for the ISF:8 and 11.9 (SD 7.6) for the ISF:16 (p < 0.001). Statistically significant associations were found between oral abnormalities and the subscales of the ISF:8 and ISF:16 (p < 0.05). Both test-retest stability and internal consistency, as measured by the intra-class correlation coefficient (ICC) (ISF:8 = 0.98 and ISF:16 = 0.97) and Cronbach's alpha (ISF:8 = 0.70 and ISF:16 = 0.84) proved to be adequate. Construct validity was confirmed from the correlation between the short form scores and oral health and overall well-being ratings. The score on the short forms of the CPQ11-14 was able to discriminate between different oral conditions. Criterion validity was satisfactory (p < 0.05). Conclusion: The Brazilian versions of CPQ11-14 ISF:8 and ISF:16 have satisfactory psychometric properties, similar to those of the original instrument. [ABSTRACT FROM AUTHOR]

Melon residues were used to produce cellulases (EC 3·.1·4) and to investigate the prebiotic effect in vitro. Peel and seed flours were characterized regarding contents of dietetic fibre, lipids, total protein, cellulose, hemicelluloses and lignin. A kinetic study was carried out in bacteriological greenhouse during 24, 48, 72, 96, 120, 144 and 168 h. In melon seed flour, cellulose at 350 g·kg-1, hemicellulose at 79 g·kg-1 and lignin at 24 g·kg-1 were determined. Results showed, in terms of dietetic fibre and high protein contents, that both seeds and peels have good nutritional value. The highest content of lipids, 246 g·kg-1, was found in seeds, compared to only 36·3 g·kg-1 in the peel. The best activity for carboxymethyl cellulase in 144 h of fermentation was 1.045 U·g-1, while the activity with filter paper after 96 h of cultivation was 0·190 U·g-1. Melon seed flour demonstrated to be a good substrate for the growth of bifidobacteria with 8 h for fermentation, and it was tolerant to the action of bile salts at 8 h of fermentation. Melon residues can be taken as a potential prebiotic ingredient and a source of cellulolytic enzymes. [ABSTRACT FROM AUTHOR]

Abstract Background Traumatic dental injury (TDI) could have physical and psychosocial consequences for children. Thus, it is important to measure the impact of TDI on the quality of life of children (QoL). The aim of the present study was to investigate the association between treated/untreated TDI and the impact on the quality of life of 11-to-14-year-old Brazilian schoolchildren. Methods A cross-sectional study was carried out involving 1612 male and female schoolchildren aged 11 to 14 years attending public and private elementary schools in the city of Belo Horizonte, Brazil. A multi-stage sampling technique was adopted to select the children. Three calibrated examiners used the Andreasen classification for the diagnosis of TDI. Oral health-related quality of life was assessed using the Brazilian version of the Child Perceptions Questionnaire (CPQ11-14) - Impact Short Form (ISF:16), composed of 16 items and self-administered by all children. Other oral conditions (dental caries and malocclusion) and the Social Vulnerability Index were determined and used as controlling variables. Results Two hundred nineteen children were diagnosed with untreated TDI and 64 were diagnosed with treated TDI. There were no statistically significant associations between untreated or treated TDI and overall CPQ11-14 (Fisher = 0.368 and Fisher = 0.610, respectively). Children with an untreated TDI were 1.4-fold (95% CI = 1.1-2.1) more likely to report impact on the item "avoided smiling/laughing" than those without TDI, whereas children with a treated TDI were twofold (95% CI = 1.1-3.5) more likely to report impact on the item "other children asked questions" than those without TDI. Conclusions Neither treated nor untreated TDI was associated with oral symptoms, functional limitations or emotional wellbeing. However, children with a TDI in the anterior teeth experienced a negative impact on social wellbeing, mainly with regard to avoiding smiling or laughing and being concerned about what other people may think or say.

Abstract Background The need to evaluate the impact of oral health has led to the development of instruments for measuring oral health-related quality of life (OHQoL). One such instrument is the Child Perceptions Questionnaire (CPQ11–14), developed specifically for 11-to-14-year-old children. As this questionnaire was considered long (37 items), shorter forms were developed with 8 (Impact Short Form: 8 – ISF:8) and 16 items (Impact Short Form: 16 – ISF:16) to facilitate use in the clinical setting and population-based health surveys. The aim of the present study was to translate and cross-culturally adapt these CPQ11–14 short forms for Brazilian Portuguese and evaluate the measurement properties of these versions for use on Brazilian children. Methods Following translation and cross-cultural adaptation, the ISF:8 and ISF:16 were tested on 136 children from 11 to 14 years of age in the city of Belo Horizonte, Brazil. The instrument was administered by a trained researcher who also performed clinical examinations. The measurement properties (i.e. criterion validity, construct validity, internal consistency reliability, test-retest reliability) were determined. Discriminant validity was tested between groups, which were divided into children with no cavities and no malocclusion; children with cavities and without malocclusion; and children with malocclusion and without cavities. Results The mean total score was 6.8 [standard deviation (SD) 4.2] for the ISF:8 and 11.9 (SD 7.6) for the ISF:16 (p < 0.001). Statistically significant associations were found between oral abnormalities and the subscales of the ISF:8 and ISF:16 (p < 0.05). Both test-retest stability and internal consistency, as measured by the intra-class correlation coefficient (ICC) (ISF:8 = 0.98 and ISF:16 = 0.97) and Cronbach's alpha (ISF:8 = 0.70 and ISF:16 = 0.84) proved to be adequate. Construct validity was confirmed from the correlation between the short form scores and oral health and overall well-being ratings. The score on the short forms of the CPQ11–14 was able to discriminate between different oral conditions. Criterion validity was satisfactory (p < 0.05). Conclusion The Brazilian versions of CPQ11–14 ISF:8 and ISF:16 have satisfactory psychometric properties, similar to those of the original instrument.

Language disorders may interfere with social integration and affectpersonal development. Beca-use the balance of the stomatognathic system can interfere with language, it is important for speech therapy and dentistry to work together, providing multidisciplinar y Healthcare., Aim: To analyze the 100 most frequently cited articles on language disorders in children and adolescents and assess the interplay with dentistry by means of a bibliometric analysis., Materials and Method: A search of the 100 most frequently cited articles up to December 2021 on language disorders was performed in the Web of Science Core Collection database. Four researchers extracted the data on number of citations, title, authors, country, year of publication, journals, study design, prevalent clinical conditions, and area of expertise. The analyses were performed using VOSviewer and Excel., Results: The total number of citations ranged from 251 to 1,431. Four articles were cited more than 1,000 times. Bishop DVM (10 articles; 3,653 citations) and Tomblin JD (10 articles; 4,261 citations) were the most frequently cited authors. The institutions with the largest number of publications were the University of Oxford/England (11%) and the University of Kansas/USA (8%). Observational study design was the most frequent (77%). Autism spectrum disorder (18%) and dyslexia (14%) were the most broadly investigated clinical conditions. Speech-language pathology (32%) was the area of expertise with the largest number of articles, and none of the top 100 studies showed interplay with dentistry., Conclusion: The 100 most widely cited articles on language disorders are mostly observational, mainly address autism spectrum disorder, and are in the field of speech-language pathology No study reported interplay with dentistry., Competing Interests: The authors declare no potential conflicts of interest regarding the research, authorship, and/or publication of this article., (Copyright© 2024 Sociedad Argentina de Investigación Odontológica.)

The Transcription Termination factor Rho is a ring-shaped, homohexameric protein that causes transcript termination by interaction with specific sites on nascent mRNAs. The process of transcription termination is essential for proper expression and regulation of bacterial genes. Although Rho has been extensively studied in the model bacteria Escherichia coli (EcRho), the properties of other Rho orthologues in other bacteria are poorly characterized. Here we present the heterologous expression and purification of untagged Rho protein from the diazotrophic environmental bacterium Azospirillum brasilense (AbRho). The AbRho protein was purified to >99% through a simple, reproducible and efficient purification protocol, a two-step chromatography procedure (affinity/gel filtration). By using analytical gel filtration and dynamic light scattering (DLS), we found that AbRho is arranged as an homohexamer as observed in the EcRho orthologue. Secondary structure and enzyme activity of AbRho was also evaluate indicating a properly folded and active protein after purification. Enzymatic assays indicate that AbRho is a RNA-dependent NTPase enzyme., (Copyright © 2022. Published by Elsevier Inc.)

Robots have been used to offset the limb weight through gravity compensation in upper body rehabilitation to delineate the effects of loss of strength and loss of dexterity, which are two common forms of post-stroke impairments. In this paper, we explored the impact of this anti-gravity support on the quality of movement during reaching and coordinated arm movements in a pilot study with two participants with chronic stroke. The subjects donned the Harmony exoskeleton which supported proper shoulder coordination in addition to providing gravity compensation. Participants had previously taken part in seven one-hour sessions with the Harmony exoskeleton, performing six sets of passive-stretching and active exercises. Pre- and post-training sessions included assessments of two separate tasks, planar reaching and a set of six coordinated arm movements, in two conditions, outside of and supported by the exoskeleton. The movements were recorded using an optical motion capture system and analyzed using spectral arc length (SPARC) and straight line deviation to quantify movement smoothness and quality. We observed that gravity compensation resulted in an increased smoothness for the subject with high level of impairment whereas compensation resulted in a reduction in smoothness for the subject with low level of impairment in the reaching task. Both participants demonstrated better coordination of the shoulder-arm joint with gravity compensation. This result motivates further studies into the role of gravity compensation during coordinated movement training and rehabilitation interventions.

Voltammetry and amperometry are inexpensive and high-performance analytical techniques. However, their lack of selectivity limits their use in complex matrices such as biological, environmental, and food samples. Therefore, voltammetric and amperometric analyses of these samples usually require time-consuming and laborious sample pretreatments. In this study, we present a simple and cost-effective approach to fabricate a miniaturized electrochemical cell that can be easily coupled to a head space-like gas extraction procedure in such a way the sample pretreatment and voltammetric detection are performed in a single step. As a proof of concept, we have used the proposed system to quantify sulfite in beverage samples after its conversion to SO 2(g) . Despite the simplicity and low cost of the proposed system, it provided good analytical performance and a limit of detection of 4.0 μmol L -1 was achieved after only 10 min of extraction. The proposed system is quite versatile since it can be applied to quantify any volatile electroactive species. Also, the proposed system provides a unique way to assess real-time extraction curves, which are essential to study and optimize new gas extraction procedures. Therefore, the approach described in this study could contribute to both applied and fundamental Analytical Chemistry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Infections and inflammation lead to a downregulation of drug metabolism and kinetics in experimental animals. These changes in the expression and activities of drug-metabolizing enzymes may affect the effectiveness and safety of pharmacotherapy of infections and inflammatory conditions., Objective: In this review, we addressed the available evidence on the effects of malaria on drug metabolism activity and kinetics in rodents and humans., Results: An extensive literature review indicated that infection by Plasmodium spp consistently decreased the activity of hepatic Cytochrome P450s and phase-2 enzymes as well as the clearance of a variety of drugs in mice (lethal and non-lethal) and rat models of malaria. Malaria-induced CYP2A5 activity in the mouse liver was an exception. Except for paracetamol, pharmacokinetic trials in patients during acute malaria and in convalescence corroborated rodent findings. Trials showed that, in acute malaria, clearance of quinine, primaquine, caffeine, metoprolol, omeprazole, and antipyrine is slower and that AUCs are greater than in convalescent individuals., Conclusion: Notwithstanding the differences between rodent models and human malaria, studies in P. falciparum and P. vivax patients confirmed rodent data showing that CYP-mediated clearance of antimalarials and other drugs is depressed during the symptomatic disease when rises in levels of acute-phase proteins and inflammatory cytokines occur. Evidence suggests that inflammatory cytokines and the interplay between malaria-activated NF-kB-signaling and cell pathways controlling phase 1/2 enzyme genes transcription mediate drug metabolism changes. The malaria-induced decrease in drug clearance may exacerbate drug-drug interactions, and the occurrence of adverse drug events, particularly when patients are treated with narrow-margin-of-safety medicines., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Objectives: The presence of autoimmune events were recorded in patients receiving immune checkpoint inhibitors., Materials & Methods: Retrospective study in patients receiving immune checkpoint inhibitors (ICIs) during the period of 2012-2019., Results: A total of 554 patients received ICIs of which 123 developed an immune related adverse event. Twenty one (17%) with toxicity were identified as having a pre-existing autoimmune disease and 88 required treatment with corticosteroids or hormone replacement. Thirty two (26%) out of 123 had to temporarily discontinue ICIs due to autoimmune manifestations. Endocrine and skin manifestations were the most prevalent immune disorders in our cohort. In melanoma better efficacy was seen in patients with immune toxicity., Conclusion: Autoimmune diseases appear in patients receiving ICIs in this real world experience. Our results differ from other series on the frequency of autoimmunity. Complete discontinuation of ICIs due to autoimmunity was rare., Competing Interests: Financial & competing interests disclosure The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties. No writing assistance was utilized in the production of this manuscript., (© 2020 Morton Scheinberg.)

Harmony is a bimanual upper-limb exoskeleton designed for post-stroke rehabilitation. It moves the subject's shoulders and arms through their entire ranges of motion while maintaining natural coordination, is capable of force/torque control of each joint, and is equipped with sensors to measure motions and interaction forces. With these capabilities Harmony has the potential to assess motor function and create individualized therapy regimens. As a first step, five stroke survivors underwent rehabilitation sessions practicing multijoint movements with the device. Each participant performed a total of 1130 motions over seven hours of therapy with no adverse effects reported by participants or the attending therapist, supporting the suitability of Harmony for use in a clinical setting. Donning and doffing time averaged 3.5 minutes and decreased with therapist experience. Reported levels of stress, anxiety, and pain indicate that the Harmony safely assisted in the completion of the trained movements and has great potential to motivate and engage patients. We developed a novel methodology for assessing coordination capability and results from the study indicate that Harmony can enable therapists to identify neuromuscular weakness and maladaptive coordination patterns and develop targeted interventions to address these aspects of upper-limb function. The results suggest Harmony's feasibility and show promising improvements, motivating future study to gain statistical support.

Introduction: Accidental damage to the inferior alveolar nerve is of concern during endodontic treatment of the posterior mandible teeth, and a careful evaluation of the relationship between the apices and the mandibular canal (MC) is mandatory to prevent iatrogenic complications. This study aimed to assess the distance between the root apices of posterior teeth and the MC and the bone density in such areas., Methods: One thousand retrospective cone-beam computed tomographic scans were randomly selected from a private oral radiologic clinic database. The images were acquired with a limited field of view and analyzed by 2 evaluators who measured the shortest distance between the root apices and the upper cortical bone of the MC using Dental Slice software (Bioparts Biomedical Prototyping, Brasília, DF, Brazil). In addition, bone density was estimated using the tool "profile line of Hounsfield" and expressed in Hounsfield units., Results: The distal root of the right second molars was the closest to the MC (mean = 3.41 mm), and the right first premolar root was the farthest from the MC (mean = 5.87 mm), respectively. The root apices were closer to the MC in women than in men (P < .05). In 1.79% of the cases, the roots were in contact or invading the MC, whereas 8.35% of the root apices were close to the MC (<1.00 mm), and 89.85% were distant from the MC (>1.00 mm). In 85.55% of the cases, the bone was dense., Conclusions: The results suggest that the risk of damage to the inferior alveolar nerve is higher for second molars, especially in female patients. The higher density of the adjacent bone may act as a protective factor against nerve damage arising from endodontic procedures., (Copyright © 2019 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Background and Purpose: The endocannabinoids anandamide and 2-arachidonoylglycerol (2-AG) bind to CB 1 and CB 2 cannabinoid receptors in the brain and modulate the mesolimbic dopaminergic pathway. This neurocircuitry is engaged by psychostimulant drugs, including cocaine. Although CB 1 receptor antagonism and CB 2 receptor activation are known to inhibit certain effects of cocaine, they have been investigated separately. Here, we tested the hypothesis that there is a reciprocal interaction between CB 1 receptor blockade and CB 2 receptor activation in modulating behavioural responses to cocaine., Experimental Approach: Male Swiss mice received i.p. injections of cannabinoid-related drugs followed by cocaine, and were then tested for cocaine-induced hyperlocomotion, c-Fos expression in the nucleus accumbens and conditioned place preference. Levels of endocannabinoids after cocaine injections were also analysed., Key Results: The CB 1 receptor antagonist, rimonabant, and the CB 2 receptor agonist, JWH133, prevented cocaine-induced hyperlocomotion. The same results were obtained by combining sub-effective doses of both compounds. The CB 2 receptor antagonist, AM630, reversed the inhibitory effects of rimonabant in cocaine-induced hyperlocomotion and c-Fos expression in the nucleus accumbens. Selective inhibitors of anandamide and 2-AG hydrolysis (URB597 and JZL184, respectively) failed to modify this response. However, JZL184 prevented cocaine-induced hyperlocomotion when given after a sub-effective dose of rimonabant. Cocaine did not change brain endocannabinoid levels. Finally, CB 2 receptor blockade reversed the inhibitory effect of rimonabant in the acquisition of cocaine-induced conditioned place preference., Conclusion and Implications: The present data support the hypothesis that CB 1 and CB 2 receptors work in concert with opposing functions to modulate certain addiction-related effects of cocaine., Linked Articles: This article is part of a themed section on 8 th European Workshop on Cannabinoid Research. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v176.10/issuetoc., (© 2018 The British Pharmacological Society.)

The possibility of using the RNA interference (RNAi) mechanisms in gene therapy was one of the scientific breakthroughs of the last century. Despite the extraordinary therapeutic potential of this approach, the need for an efficient gene carrier is hampering the translation of the RNAi technology to the clinical setting. Although a diversity of nanocarriers has been described, liposomes continue to be one of the most attractive siRNA vehicles due to their relatively low toxicity, facilitated siRNA complexation, high transfection efficiency and enhanced pharmacokinetic properties. This review focuses on RNAi as a therapeutic approach, the challenges to its application, namely the nucleic acids' delivery process, and current strategies to improve therapeutic efficacy. Additionally, lipid-based nanocarriers are described, and lessons learned from the relation between biophysical properties and biological performance of the dioctadecyldimethylammonium:monoolein (DODAX: MO) system are explored. Liposomes show great potential as siRNA delivery systems, being safe nanocarriers to protect nucleic acids in circulation, extend their half-life time, target specific cells and reduce off-target effects. Nevertheless, several issues related to delivery must be overcome before RNAi therapies reach their full potential, namely target-cell specificity and endosomal escape. Understanding the relationship between biophysical properties and biological performance is an essential step in the gene therapy field., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

The hygiene hypothesis was proposed almost three decades ago. Nevertheless, its mechanism still remains with relevant controversies. Some studies defend that early exposures during childhood to microbes and parasites are key determinants to prevent allergies and autoimmune diseases; however, other studies demonstrated that these early exposures can even potentiate the clinical scenario of the diseases. Based on several studies covering the influences of microbiome, parasites, related theories and others, this review focuses on recent advances in the hygiene hypothesis field. In addition, the main immunological mechanisms underlying the hygiene hypothesis are also discussed. We also strongly encourage that researchers do not consider the hygiene hypothesis as a theory based strictly on hygiene habits, but a theory combining diverse influences, as illustrated in this review as the hygiene hypothesis net., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: The 2 main forms of thrombotic microangiopathy (TMA) are thrombotic thrombocytopenic purpura (TTP) and atypical hemolytic uremic syndrome (aHUS). Deficiency of ADAMTS13 and dysregulation of the complement pathway result in TTP and aHUS, respectively; however, overlap of their clinical characteristics makes differential diagnosis challenging., Objectives and Methods: We aimed to develop a TMA diagnosis workflow based on ADAMTS13 activity and screening of ADAMTS13 and complement genes using a custom next-generation sequencing (NGS) gene panel., Patients: For this, from a cohort of 154 Portuguese patients with acute TMA, the genotype-phenotype correlations were analyzed in 7 hereditary TTP (ADAMTS13 activity <10%, no inhibitor), 36 acquired TTP (ADAMTS13 activity <10%, presence of an inhibitor), and in 34 presumable aHUS., Results: In total, 37 different rare variants, 8 of which novel (in ADAMTS13 , CFH , and CD46 ), were identified across 7 genes. Thirteen TTP patients were homozygous (n=6), compound heterozygous (n=2), and heterozygous (n=5) for 11 ADAMTS13 variants (6 pathogenic mutations). Among the 34 aHUS patients, 17 were heterozygous for 23 variants in the different complement genes with distinct consequences, ranging from single pathogenic mutations associated with complete disease penetrance to benign variants that cause aHUS only when combined with other variants and/or CFH and CD46 risk haplotypes or CFHR1-3 deletion., Conclusions: Our study provides evidence of the usefulness of the NGS panel as an excellent technology that enables more rapid diagnosis of TMA, and is a valuable asset in clinical practice to discriminate between TTP and aHUS.

Portal hypertension (PHT) is a major consequence of any chronic liver disease and it is the main cause of complications in patients with cirrhosis. Measurement of hepatic vein pressure gradient is considered the gold standard for PHT assessment, together with its diagnosis and prognosis relevance. Even though hepatic vein pressure gradient measurement is a safe procedure, it is still considered an invasive technique and not widely available. There is thus a need for noninvasive methods that can predict the progression of PHT as well as the presence and the risk of complications related to esophageal varices. This review aimed to discuss the noninvasive markers used in the assessment of PHT and detection of high-risk esophageal varices in patients with liver cirrhosis. We focus on the main biomarkers, particularly those used in the routine assessment of chronic liver disease, and the physical methods that use tissue elastography as a diagnosis tool.

This work evaluates the environmental performance of using pulp and paper sludge as feedstock for the production of second generation ethanol. An ethanol plant for converting 5400 tons of dry sludge/year was modelled and evaluated using a cradle-to-gate life cycle assessment approach. The sludge is a burden for pulp and paper mills that is mainly disposed in landfilling. The studied system allows for the valorisation of the waste, which due to its high polysaccharide content is a valuable feedstock for bioethanol production. Eleven impact categories were analysed and the results showed that enzymatic hydrolysis and neutralisation of the CaCO3 are the environmental hotspots of the system contributing up to 85% to the overall impacts. Two optimisation scenarios were evaluated: (1) using a reduced HCl amount in the neutralisation stage and (2) co-fermentation of xylose and glucose, for maximal ethanol yield. Both scenarios displayed significant environmental impact improvements., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Introduction: Inherited protein C (PC) deficiency is a well-known risk factor for venous thrombosis (VT). Plasma PC levels are reliable in moderate to severe deficiencies; however, in mildly deficient individuals, the levels may overlap with those considered normal. Genetic studies of PROC, which encodes PC, could help identify carriers; genome-wide association studies (GWAS) have shown that approximately 50% of phenotypic variation in PC deficiency is caused by the cumulative effects of mutations in several other loci, namely in the PROCR., Patients and Methods: With the main objective of determining the genotype/phenotype correlation in 59 Portuguese individuals from 26 unrelated families with history of thrombosis and repeatedly low/borderline PC plasma levels, we conducted a molecular study by direct sequencing of PROC; PROC promoter haplotypes and PROCR c.4600A>G polymorphism (rs867186), which are known to influence plasma PC concentrations, were also screened., Results: Twelve different PROC mutations were identified, one of them not previously reported, p.Cys105Arg. The mutation types and locations as well as haplotype combinations correlated with the phenotypic severity. The most frequent mutation, p.Arg199X, correlated with the CGTC haplotype and was identified in nine families containing patients with higher numbers of VT episodes. This mutation in homozygous individuals for the CGTC haplotype is a significant risk factor for VT in Portuguese., Conclusion: These genetic family studies allowed the identification of the unknown carriers and individuals at a higher thrombotic risk within each family, thus permitting the evaluation of the need for prophylactic measures, particularly in at-risk situations., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

While the delivery of small interfering RNAs (siRNAs) is an attractive strategy to treat several clinical conditions, siRNA-nanocarriers' stability after intravenous administration is still a major obstacle for the development of RNA-interference based therapies. But, although the need for stability is well recognized, the notion that strong stabilization can decrease nanocarriers' efficiency is sometimes neglected. In this work we evaluated two stealth functionalization strategies to stabilize the previously validated dioctadecyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. The nanocarriers were pre- and post-pegylated, forming vectors with different stabilities in biological fluids. The stealth nanocarriers' behavior was tested under biological mimetic conditions, as the production of stable siRNA-lipoplexes is determinant to achieve efficient intravenous siRNA delivery to cancer cells. Upon incubation in human serum for 2h, by fluorescence Single Particle Tracking microscopy, PEG-coated lipoplexes were found to have better colloidal stability as they could maintain a relatively stable size. In addition, using fluorescence fluctuation spectroscopy, post-pegylation also proved to avoid siRNA dissociation from the nanocarriers in human serum. Concomitantly it was found that PEG-coated lipoplexes improved cellular uptake and transfection efficiency in H1299 cells, and had the ability to silence BCR-ABL, affecting the survival of K562 cells. Based on an efficient cellular internalization, good silencing effect, good siRNA retention and good colloidal stability in human serum, DODAB:MO (2:1) siRNA-lipoplexes coated with PEG-Cer are considered promising nanocarriers for further in vivo validation., Statement of Significance: This work describes two stealth functionalization strategies for the stabilization of the previously validated dioctadecyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. These nanocarriers are capable of efficiently incorporating and delivering siRNA molecules to cells in order to silence genes whose expression is implicated in a pathological condition. The main objective was to functionalize these nanocarriers with a coating conferring protection to siRNA in blood without compromising its efficient delivery to cancer cells, validating the potential of DODAB:MO (2:1) siRNA-lipoplexes as therapeutic vectors. We show that the stealth strategy is determinant to achieve a stable and efficient nanocarrier, and that DODAB:MO mixtures have a very promising potential for systemic siRNA delivery to leukemic cells., (Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Dioctadecyldimethylammonium bromide (DODAB):1-monooleoyl-rac-glycerol (MO) cationic liposomes were reported as a promising alternative to common transfection agents, showing superior effectiveness on the transfection of the 293T mammalian cell line with pSV-β-gal plasmid DNA. The study of DODAB:MO aggregates in the absence of DNA has indicated that their morphology depends on the balance between DODAB's tendency to form bilayer structures and MO's propensity to form inverted non-lamellar structures. Other parameters, such as the temperature have proved to be crucial in the definition of the morphology of the developed nanocarrier. Therefore, in this work, a step forward to the current gene carrier system will be given by studying the effect of the tunable parameters (incubation temperature and MO content) on the structure of pDNA:DODAB:MO lipoplexes. More importantly, the implications that these tunable parameters could have in terms of lipoplex transfection efficiency will be investigated. Dynamic light scattering (DLS), zeta (ζ) potential, cryo-transmission electron microscopy (cryo-TEM) and ethidium bromide (EtBr) exclusion were used to assess the formation, structure and destabilization of pDNA:DODAB:MO lipoplexes at DODAB molar fractions of (1:1) and above equimolarity (2:1, 4:1) prepared at incubation temperatures from 25 to 50°C. Experimental results indicate that pDNA:DODAB:MO's structure is sensitive to the lipoplex incubation temperature, resulting in particles of distinct size, superficial charge and structure. These variations are also visible on the complexation dynamics of pDNA, and subsequent release upon incubation with the model proteoglycan heparin (HEP), at 25 and 50°C. Increase in temperature leads to re-organization of DODAB and MO molecules within the liposomal formulation, causing a positive charge re-localization in the lipoplex surface, which not only alters its structure but also its transfection efficiency. Altogether, these results confirm that in the DODAB:MO carriers, an increase in the incubation temperature has a similar effect on aggregate morphology as the observed with an increase in MO content. This conclusion is extended to the pDNA:DODAB:MO lipoplexes morphology and subsequent transfection efficiency defining new strategies in lipoplexes preparation that could be used to modulate the properties of other lipid formulations for nonviral gene delivery applications., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Introduction: The pharmacological assessment of the factors for gastric protection of a test substance should involve experimental models that can determine the involvement of cytoprotective factors, as well as their influence on the secretion of hydrochloric acid. The original protocol of pylorus ligation in rats proposed by Shay et al. in 1945, still in use today, provides a latency time of 240 min without considering the effect of postoperative pain in the mechanisms of peptic ulcer. This paper proposes a modification of this experimental protocol by eliminating the pain throughout the postoperative period, as a refinement of the test with consequent improvement of the pharmacological response., Methods: Adult male Wistar/Uni rats underwent surgical ligation of the pylorus and were kept anesthetized throughout the experimental period (4h) in contrast to the other experimental groups that followed the original protocol proposed by Shay et al., 1945., Results: We were able to determine effective doses for a positive control, as well as of a variety of secretagogues in the new experimental protocol proposed., Discussion: The suppression of post-surgical pain, through the use of anesthesia throughout the experimental period, brought several benefits for the study of gastric acid secretion, rendering a more homogeneous pharmacologic response in non-inbred animals, thus being an effective experimental procedure., (Copyright © 2012 Elsevier Inc. All rights reserved.)

It has been reported that malaria reduces cytochrome-P450 (CYP) content and monooxygenase activities in the mammalian host liver. The mechanism by which malaria modulates CYP activities, however, remains unclear. In this study we found that activities of ethoxy- and benzyloxy-resorufin-O-dealkylases, p-nitrophenol-hydroxylase and erythromycin-N-demethylase (mediated by CYP1A, 2B, 2E1 and 3A, respectively) were depressed, while uridine-glucuronosyl-transferase (a phase 2 enzyme) was unaltered in liver microsomes of Plasmodium berghei-infected (parasitemia >20%) male Swiss Webster mice. Prolongation of midazolam sleeping time and a slower clearance of chlorzoxazone were also noted in infected mice. Reductions of hepatic levels of CYP1A2, 2E1 and 3A11 mRNAs indicated that malaria downregulated these CYP-mediated activities at a pre-translational level.

The induction of cytochrome P4501A-mediated activity (e.g. ethoxyresorufin-O-deethylation, EROD) has been used as a biomarker for monitoring fish exposure to AhR-receptor ligands such as polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB) and polychlorinated dibenzo-dioxins/furans (PCDD/Fs). In this study we found that hepatic EROD is induced in fish ("Nile tilapia", Oreochromis niloticus and "acará", Geophagus brasiliensis) from the Guandu River (7-17-fold) and Jacarepaguá Lake (7-fold), Rio de Janeiro, Brazil. Since both cichlid fish are consumed by the local population and the Guandu River is the main source of the drinking water supply for the greater Rio de Janeiro metropolitan area, pollution by cytochrome P4501A-inducing chemicals is a cause for concern and should be further investigated in sediments, water and biota. We additionally showed that EROD activity in the fish liver post-mitochondrial supernatant-simpler, cheaper and less time consuming to prepare than the microsomal fraction-is sufficiently sensitive for monitoring purposes.

Malaria has been reported to modulate the activity of cytochrome-P450 enzymes (CYP). Since CYPs are involved both in the activation and detoxication of xenobiotics, we investigated whether malaria would modify the effects of chemical carcinogens in the bone-marrow micronucleus assay. Female C57BL6 mice were infected with Plasmodium berghei (ANKA) and treated (ip route) with cyclophosphamide (CPA, 25 mg/kg body weight), 7,12-dimethylbenz[a]anthracene (DMBA, 50mg/kg body weight) or ethyl methanesulfonate (EMS, 150 mg/kg body weight), on post-infection days 9-12 when parasitemia was > or =9% of RBC. Controls were age-paired non-infected mice. Bone marrows were sampled at 24 and 48 h (CPA), 24 h (EMS) or 48 h (DMBA) after treatment. The background incidence of polychromatic erythrocytes with micronuclei (MN-PCE) in malaria-infected mice was approximately twofold the background incidence in non-infected controls. Effects of indirect clastogens (CPA and DMBA) in the micronucleus assay were attenuated while the effect of EMS, a direct clastogen, was enhanced by infection. In a separate experiment, malaria was shown to decrease activities of ethoxy-(EROD, a marker for CYP1A) and benzyloxy-(BROD, CYP2B) resorufin-O-dealkylases in liver microsomes. The foregoing findings are consistent with the hypothesis that malaria-caused attenuation of genotoxicity arose from a down modulation of CYP isoforms that convert CPA (CYP2B) and DMBA (CYP1A) into their active metabolites.

It has been reported that malaria infection impairs hepatic drug clearance and causes a down-regulation of CYP-mediated monooxygenase activities in rodents and humans. In the present study, we investigated the effects of Plasmodium berghei infection on the activity of liver monooxygenases in female DBA/2 and C57BL/6 mice. In both mouse strains, P. berghei infection decreased activities mediated by CYP1A (EROD: DBA/2 65.3%, C57BL/6 44.7%) and 2B (BROD: DBA/2 64.3%, C57BL/6 49.8%) subfamily isoforms and increased activities mediated by 2A5 (COH: DBA/2 182.4%, C57BL/6 148.5%) and 2E1 (PNPH: DBA/2 177.8%, C57BL/6 128.5%) isoforms as compared to non-infected controls. Since malaria infection also produced an increase in ALT (273.1%) and AST (354.1%) activities in the blood serum, our findings are consistent with the view that CYP2A5 activity is induced by liver injury. An almost generalized depression of CYP-mediated activities has been found with numerous infections and inflammatory stimuli but an induction of CYP2A5 had been previously noted only in some viral hepatitis and trematode (liver fluke) infections.

Schistosoma mansoni has been reported to cause a downregulation of hepatic cytochrome P450 activities after granulomas are formed around worm eggs harbored in the mouse liver. Only a few studies, however, provided data on the activity of xenobiotic-biotransaformation enzymes in the early phase of S. mansoni infection. In this study, we evaluated the alterations of liver microsomal enzymes during early infection (post-infection days, PIDs, 15 and 30) when granulomas are not found in the mouse liver yet. Swiss Webster (SW) and DBA/2 mice of either sex were infected with 100 S. mansoni cercariae on postnatal day 10. Levels of total-CYPs and activities of alkoxyresorufin-O-dealkylases (EROD, MROD, PROD and BROD), N-nitrosodimethylamine-N-demethylase (NDMA-d), coumarin 7-hydroxylase (COH, DBA/2 only) and UDP-glucuronosyltransferase (UGT) were measured in liver microsomes from mice killed on PIDs 15 and 30. Age-matched (sham-infected) mice of the same sex and strain were used as controls. Neither total-CYP levels nor microsomal enzyme activities were altered in SW and DBA/2 mice on PID 15. On PID 30, total-CYP levels, and COH, PROD and UGT activities remained unaltered, while gender- and strain-specific minor changes of EROD, MROD, BROD and NDMA-d (i.e., increase in SW and reduction in DBA/2) were found. In conclusion, our results suggest that, contrasting to a consistent and almost generalized downregulation of CYPs in chronic schistosomiasis, alterations of hepatic CYPs in early (acute) infection are isoform and mouse's gender and strain specific.

The activity of fish monooxygenases has been extensively used as a monitoring tool to detect contamination of water bodies by cytochrome P450-inducing agents. In this study we evaluated the activities of ethoxy- (EROD), methoxy- (MROD) and pentoxy- (PROD) resorufin-O-dealkylases in the liver of Nile tilapias (Oreochromis niloticus) collected at the Guandu river, at a reference clean site (Lake 1) and at two other sampling sites (Lakes 2 and 3) in Rio de Janeiro state, Brazil. Alkoxyresorufin-O-dealkylases were measured fluorimetrically in the hepatic S9 fraction. EROD (17.7-fold), MROD (14.2-fold) as well as PROD activities were considerably higher in tilapias from Guandu river. A moderate increase of EROD (5.0-fold) and MROD (5.4-fold) was also found in tilapias from Lake 3. These findings suggest that Guandu river watershed, the main source of urban drinking water supply in Rio de Janeiro, is polluted with CYP1A-inducing xenobiotics. Furthermore, we also found a good linear relationship between EROD and MROD, a finding that agrees with the hypothesis that the two reactions are catalysed by the same CYP1A isoform in O. niloticus.

How to handle Western blot (WB) seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2) constitutes a challenge for blood banks and families. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA) reactive individuals from the hematological center (HEMOCE) of Fortaleza (Brazil), examining their serological (WB) and molecular (PCR) diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22%) were positive and 32 (78%) were negative for HTLV-1/2. The demographic analysis indicated a trend towards a predominance of males among the seroindeterminate individuals and females in the seropositive ones. The seroindeterminate individuals were younger than the seropositive ones. We did not find any association of these conditions with syphilis, Chagas disease or HIV or hepatitis, and with risk factors such as breast-feeding, blood transfusion, STD (syphilis) and IDU.