Can rapid maxillary expansion affect speech sound production in growing patients? A systematic review.

Rapid maxillary expansion (RME) may change speech sound parameters due to the enlargement of oral and nasal cavities. This study aimed to systematically review the current evidence on speech changes as a side effect of RME. An electronic search was conducted in nine databases, and two of them accessed the 'grey literature'. The eligibility criteria included clinical studies assessing orthodontic patients with maxillary transverse deficiency and the relationship with speech alterations without restricting publication year or language. Only interventional studies were included. The JBI Critical Appraisal Tool assessed the risk of bias. The initial search provided 4853 studies. Seven articles (n = 200 patients) met the inclusion criteria and were analysed. The primary source of bias was the absence of a control group in four studies. RME altered speech production by changing vowel fundamental frequency and fricative phoneme formant frequency. Shimmer and jitter rates changed in one and two studies, respectively. Two studies presented deterioration during orthodontic treatment, but speech improved after appliance removal. Despite the limited evidence, RME affects speech during and after treatment.

Water Leakage Pathway Leads to Internal Hydration of the p53 Core Domain.

The gene encoding the p53 tumor suppressor protein is the most frequently mutated oncogene in cancer patients; yet, generalized strategies for rescuing the function of different p53 mutants remain elusive. This work investigates factors that may contribute to the low inherent stability of the wild-type p53 core domain (p53C) and structurally compromised Y220C mutant. Pressure-induced unfolding of p53C was compared to p63C, the p53 family member with the highest stability, the engineered superstable p53C hexamutant (p53C HM), and lower stability p53C Y220C cancer-associated mutant. The following pressure unfolding values (P50% bar) were obtained: p53C 3346, p53C Y220C 2217, p53C HM 3943, and p63C 4326. Molecular dynamics (MD) simulations revealed that p53C Y220C was most prone to water infiltration, followed by p53C, whereas the interiors of p53C HM and p63C remained comparably dry. A strong correlation (r2 = 0.92) between P50% and extent of interior hydration was observed. The pathways of individual water molecule entry and exit were mapped and analyzed, revealing a common route preserved across the p53 family involving a previously reported pocket, along with a novel surface cleft, both of which appear to be targetable by small molecules. Potential determinants of propensity to water incursion were assessed, including backbone hydrogen bond protection and combined sequence and structure similarity. Collectively, our results indicate that p53C has an intrinsic susceptibility to water leakage, which is exacerbated in a structural class mutant, suggesting that there may be a common avenue for rescuing p53 function.

Acute effects of intracerebroventricular administration of α-ketoisocaproic acid in young rats on inflammatory parameters.

Maple Syrup Urine Disease (MSUD) is an autosomal recessive inborn error of metabolism (IEM), responsible for the accumulation of the branched-chain amino acids (BCAA) leucine, isoleucine, and valine, in addition to their α-keto acids α-ketoisocaproic acid (KIC), α-keto-ß-methylvaleric acid (KMV), and α-ketoisovaleric acid (KIV) in the plasma and urine of patients. This process occurs due to a partial or total blockage of the dehydrogenase enzyme activity of branched-chain α-keto acids. Oxidative stress and inflammation are conditions commonly observed on IEM, and the inflammatory response may play an essential role in the pathophysiology of MSUD. We aimed to investigate the acute effect of intracerebroventricular (ICV) administration of KIC on inflammatory parameters in young Wistar rats. For this, sixteen 30-day-old male Wistar rats receive ICV microinjection with 8 µmol KIC. Sixty minutes later, the animals were euthanized, and the cerebral cortex, hippocampus, and striatum structures were collected to assess the levels of pro-inflammatory cytokines (INF-γ; TNF-α, IL-1ß). The acute ICV administration of KIC increased INF-γ levels in the cerebral cortex and reduced the levels of INF-γ and TNF-α in the hippocampus. There was no difference in IL-1ß levels. KIC was related to changes in the levels of pro-inflammatory cytokines in the brain of rats. However, the inflammatory mechanisms involved in MSUD are poorly understood. Thus, studies that aim to unravel the neuroinflammation in this pathology are essential to understand the pathophysiology of this IEM.

Do Go Chasing Waterfalls: Enoyl Reductase (FabI) in Complex with Inhibitors Stabilizes the Tetrameric Structure and Opens Water Channels.

The enzyme enoyl-ACP reductase (FabI) is the limiting step of the membrane's fatty acid biosynthesis in bacteria and a druggable target for novel antibacterial agents. The FabI active form is a homotetramer, which displays the highest affinity to inhibitors. Herein, molecular dynamics studies were carried out using the structure of FabI in complex with known inhibitors to investigate their effects on tetramerization. Our results suggest that multimerization is essential for the integrity of the catalytic site and that inhibitor binding enables the multimerization by stabilizing the substrate binding loop (SBL, L:195-200) coupled with changes in the H4/5 (QR interface). We also observed that AFN-1252 (naphtpyridinone derivative) promotes unique conformational changes affecting monomer-monomer interfaces. These changes are induced by AFN-1252 interaction with key residues in the binding sites (Ala95, Tyr146, and Tyr156). In addition, the analysis of water trajectories indicated that AFN-1252 complexes allow more water molecules to enter the binding site than triclosan and MUT056399 complexes. FabI-AFN-1252 simulations show accumulation of water molecules near the Tyr146/147 pocket, which can become a hotspot to the design of novel FabI inhibitors.

Pulp changes from rapid maxillary expansion: A systematic review.

INTRODUCTION: This study aimed to systematically review the current evidence on the occurrence of pulp changes as side effects from orthopaedic rapid maxillary expansion (ORME) or surgically assisted rapid maxillary expansion (SARME). METHODS: An electronic search was performed in eleven databases. The eligibility criteria included clinical studies assessing vitality, sensibility or dimensions of the pulp chamber of permanent teeth before and after ORME or SARME, without restrictions on publication year or language. The risk of bias was analysed with the NIH 'Quality Assessment Tool for Before-After (Pre-Post) Studies with No Control Group' and the 'JBI for quasi-experimental studies' tool. The GRADE tool was used to assess the certainty of evidence. RESULTS: The initial search resulted in 1,197 records, from which only seven before-after studies were included. There was a change in the pulpal blood flow (PBF) of maxillary incisors and canines up to 5 days after SARME, which gradually returned after 7 days to 3 months. After ORME, one study observed an increased PBF and one study observed a reduced PBF, which gradually returned after the end of expansion. Two studies observed that both ORME and SARME caused temporary changes in pulp sensibility. Three studies observed a significant reduction in the pulp chamber after ORME or SARME. The outcomes presented a very low certainty of evidence. CONCLUSIONS: Although limited, the evidence shows that ORME and SARME caused temporary changes in pulp vitality and sensibility, with the possibility of inducing a reduction in pulp chamber dimensions.

Antarctic environments as a source of bacterial and fungal therapeutic enzymes.

Microbial therapeutic enzymes are the protagonists in the pharmacological treatment of different human diseases. The intrinsic enzymatic characteristics, such as high affinity and specificity to the corresponding substrate, enable effective therapies, with minimal adverse effects and complete remission. However, immunogenicity, short half-life, low enzymatic yield, and low selectivity regarding available enzyme drugs are currently the main obstacles to their development and the broad adherence to therapeutic protocols. By harboring adapted and still unexplored microbial life, environments of extreme conditions, such as Antarctica, become especially important in the prospecting and development of new enzymatic compounds that present higher yields and the possibility of genetic improvement. Antarctic microorganisms have adaptation mechanisms, such as more fluid cell membranes, production of antifreeze proteins and enzymes with more malleable structures, more robust, stable, selective catalytic sites for their respective substrates, and high antioxidant capacity. In this context, this review aims to explore enzymes synthesized by bacteria and fungi from Antarctica as potential drug producers, capable of providing therapeutic efficacy, less adverse effects, and lower production costs with highlight to L-Asparaginase, collagenase, superoxide dismutase and ribonucleases. In addition, this review highlights the unique biotechnological profile of these Antarctic extremophile microorganisms.

Antarctic organisms as a source of antimicrobial compounds: a patent review.

Currently, antimicrobial resistance has become a global public health problem, which has made the need for new antimicrobial compounds to deal with resistant infections an emergency. However, environments that once offered so many innovative molecules, now already exhaustively exploited, do not meet this need. In this context, a geographically isolated, under-explored and extreme environment, such as Antarctica, which holds organisms with unique physiological and biochemical characteristics, assumes great importance as a potential source of new compounds with antimicrobial activity. In this patent review, we investigate the state of technological development in the field of antimicrobial compounds obtained from Antarctic organisms, highlighting the main countries and researchers active in the field, the species utilized, the compounds obtained, and their possible therapeutic applications. As results, few patent documents were found, however they encompass a wide diversity of compounds and species, indicating a great antimicrobial potential present in Antarctic biota, including compounds active against the most important human pathogenic microorganisms, such as including methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus spp. and multi-resistant Mycobacterium tuberculosis. Furthermore, due to the increasing trend in patent applications, a significant rise in the number of patents in this area is expected in the coming years.

Effects of low-level laser therapy on reducing pain, edema, and trismus after orthognathic surgery: a systematic review.

The purpose of this study was to systematically review the scientific evidence of the effect of low-level laser therapy (LLLT) on the perception of pain, edema, and trismus after orthognathic surgery. The literature was searched in 11 databases (MedLine via PubMed, Scopus, LILACS, SciELO, Embase, Web of Science, Cochrane Library, and LIVIVO, OpenGrey, OADT, and OpenThesis), without restriction of publication year or language. This search aimed to identify randomized clinical trials comparing low-level laser therapy and placebo for controlling pain, edema, and trismus after orthognathic surgeries. Two reviewers extracted the data and assessed the individual risk of bias of the eligible studies using the Cochrane Collaboration Risk of Bias Tool (RoB, version 2.0). The initial search resulted in 808 articles, from which only five (total of 190 participants) were included in the qualitative synthesis. The studies were published from 2014 to 2020. Two presented a low risk of bias + in the mean mouth opening of all patients subjected to bimaxillary surgery who received LLLT. However, the other study found a significant difference in maximum mouth opening in the LLLT group at 14, 30, and 60 days after surgery. Based on limited evidence, LLLT was presented as an auxiliary tool for reducing pain and trismus after surgery. However, the reduction of edema is controversial due to the absence of measuring standardization.

Antibacterial and modulatory activities of ß-cyclodextrin complexed with (+)-ß-citronellol against multidrug-resistant strains.

The present study aimed to investigate the antibacterial and modulatory activities of (+)-ß-citronellol (ßCT), ß-cyclodextrin (ß-CD), and their complex ßCT/ß-CD and characterize them using infrared spectroscopy. Infrared spectra were recorded in the 750-4000 cm-1 region. The antibacterial effects of these compounds and their modulatory-antibiotic activities were determined using the minimum inhibitory concentration (MIC) test. Signatures of these pure compounds were detected in the infrared spectrum of the ßCT/ß-CD complex. The MIC of the ßCT/ß-CD complex against the tested strains was found to be 1024 µg/mL. The antagonistic and synergistic effects of these compounds were also observed using the modulation tests. ßCT or ß-CD alone did not exhibit any direct antibacterial activity. However, the ßCT/ß-CD complex in combination with gentamicin showed a synergistic effect against E. coli.

Biophysical characterization of p53 core domain aggregates.

Aggregation is the cause of numerous protein conformation diseases. A common facet of these maladies is the transition of a protein from its functional native state into higher order forms, such as oligomers and amyloid fibrils. p53 is an essential tumor suppressor that is prone to such conformational transitions, resulting in its compromised ability to avert cancer. This work explores the biophysical properties of early-, mid-, and late-stage p53 core domain (p53C) aggregates. Atomistic and coarse-grained molecular dynamics (MD) simulations suggest that early- and mid-stage p53C aggregates have a polymorphic topology of antiparallel and parallel ß-sheets that localize to the core amyloidogenic sequence. Both topologies involve similar extents of interstrand mainchain hydrogen bonding, while sidechain interactions could play a role in regulating strand orientation. The free energy difference between the antiparallel and parallel states was within statistical uncertainty. Negative stain electron microscopy of mature fibrils shows a wide distribution of fiber widths, indicating that polymorphism may extend to the quaternary structure level. Circular dichroism of the fibrils was indicative of ß-sheet rich structures in atypical conformations. The Raman spectrum of aggregated p53C was consistent with a mixture of arranged ß-sheets and heterogeneous structural elements, which is compatible with the MD findings of an ordered ß-sheet nucleus flanked by disordered structure. Structural polymorphism is a common property of amyloids; however, because certain polymorphs of the same protein can be more harmful than others, going forward it will be pertinent to establish correlations between p53C aggregate structure and pathology.

Human acute Chagas disease: changes in factor VII, activated protein C and hepatic enzymes from patients of oral outbreaks in Pará State (Brazilian Amazon).

Oral transmission of Chagas disease has been increasing in Latin American countries. The present study aimed to investigate changes in hepatic function, coagulation factor levels and parasite load in human acute Chagas disease (ACD) secondary to oral Trypanosoma cruzi transmission. Clinical and epidemiological findings of 102 infected individuals attended in the State of Pará from October 2013 to February 2016 were included. The most common symptoms were fever (98%), asthenia (83.3%), face and limb edema (80.4%), headache (74.5%) and myalgia (72.5%). The hepatic enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of 30 ACD patients were higher compared with controls, and this increase was independent of the treatment with benznidazole. Moreover, ACD individuals had higher plasma levels of activated protein C and lower levels of factor VII of the coagulation cascade. Patients with the highest parasite load had also the most increased transaminase levels. Also, ALT and AST were associated moderately (r = 0.429) and strongly (r = 0.595) with parasite load respectively. In conclusion, the present study raises the possibility that a disturbance in coagulation and hepatic function may be linked to human ACD.

Implementation of dental specialty centers: a descriptive analysis of the current status in the Brazilian territory.

This study aimed to assess the current implementation status of Dental Specialty Centers (Centros de Especialidades Odontológicas - CEO) in Brazil. The sample included CEOs implemented up to November 2015 in the 27 Brazilian federative units. Data were obtained directly from the database of the Informatics Department of the Brazilian Unified Health System, according to the National Registry of Health Facilities (NRHF) of Dental Specialty Centers of all Brazilian regions. Primary care data were also collected from the cities with implemented CEOs, including coverage status of the Family Health Strategy (FHS) and number of Oral Health Teams (OHT) I and II, at 2 collection periods (January 2006 and November 2015). There were 1019 CEOs implemented in Brazil, which were unequally distributed among the Brazilian states, with prevalence of implementation of CEOs type II (n=503, 49.4%). The statistical analysis showed significant difference between the three types of CEO (I, II, and III) and the variables of coverage rate (FHS) and number of teams (OHT I, OHT II) at both data collection periods. Although presenting an evolutionary aspect in the implementation of CEOs, the implementation of medium-complexity care in Brazil is disorganized.

A scientific survey on 1550 cases of oral lesions diagnosed in a Brazilian referral center.

The present study aimed to perform an epidemiological survey on the prevalence rate of oral lesions diagnosed in a referral center in Brazil and to establish association between the lesions and the age and gender of the affected patients. This analytical observational study analyzed 3521 medical records of anatomopathological exams of the Pathological Anatomy and Cytopathology Laboratory of the Cancer Hospital of the state of Mato Grosso (Brazil), from December 2011 to September 2015. A total of 1550 relevant medical records was detected. Patients aging between 41 and 60 consisted of 42.84% of the sample. Males were more affected than females (60.13%). Inflammatory processess consisted of the most prevalent diagnosis (43.29%), followed by squamous cell carcinoma (6.06%), and periapical granuloma (5.23%). Oral health public policies must give major attention to Brazilian males aged between the fourth and sixth decades of life in order to improve their quality of life.

Perception, knowledge and attitudes of Brazilian orthodontists on the treatment of Class II malocclusions.

The present study aimed to assess the perception and knowledge of Brazilian orthodontists on the ideal moment to treat Class II malocclusions. Questionnaires with open, semi-open and close questions were sent by e-mail to 1653 Brazilian orthodontists. These orthodontists were registered in the Brazilian Association of Orthodontics and Dentofacial Orthopedics (ABOR). One-hundred and three (9.86%) Orthodontists replied to the questionnaires. Most of them were males (60.5%) with specialization as the highest level of professional qualification (59.9%). Most of the orthodontists (51%) reported preference for treating Class II malocclusions in late mixed dentition, followed by the early mixed dentition (29%). The age range between 10 and 12 years old figured as the most prevalent in patients searching for treatment (42.7%). Most of the patients searching for treatment were females (69.7%). Preferences for functional orthopedic appliances were reported by 35% of the orthodontists. Brazilian orthodontists revealed a trend for treating patients with Class II malocclusions in the late mixed dentition. Female patients aged from 10 to 12 years old represented the majority of patients that search for orthodontic treatment. Functional orthopedic appliances were preferred by Brazilian orthodontists for treating Class II malocclusions.

Influence of Age on Factors associated with Peri-implant Bone Loss after Prosthetic Rehabilitation over Osseointegrated Implants.

INTRODUCTION: To verify the influence of age on factors associated with peri-implant bone loss after prosthetic rehabilitation over osseointegrated implants. MATERIALS AND METHODS: This is an analytical, observational, and longitudinal study with initial 23 participants. Patients presenting with osseointegrated implants with their respective prostheses installed were included, and they could be carriers of chronic and degenerative diseases, such as diabetes, osteoporosis, hypothyroidism, cardiovascular disease (CVD), and systemic arterial hypertension. Thus, 18 participants with 57 implants were selected and followed up from 2009 to 2013. For statistical analysis, chi-square or Fisher's exact test was used for the association of systemic conditions and bone loss. Student's t-test was used for mean comparisons of age and number of total upper and lower implants. RESULTS: The average age of the sample studied was 71.05 years (65-80). The average implant per person was 3.2. Smoking had an influence on both mesial and distal bone loss, and the latter was significant (p = 0.0370). The association between bone loss and gender was also significant (p < 0.05). Moreover, male gender and upper implants were factors significantly associated with bone loss. The systemic conditions, when isolated, did not have significant influence on implant survival. CONCLUSION: Age is not a factor that, alone, contraindicates implant-rehabilitating therapy. On the contrary, smoking has a significant influence on dental implant survival. Systemic diseases, such as osteoporosis, hypothyroidism, diabetes, hypertension, and heart diseases, when controlled, are not contraindication factors. CLINICAL SIGNIFICANCE: This study is relevant for assessing peri-implant bone loss in elderly patients, right after implant installation and over time. Therefore, it was possible to verify that age is not a limiting factor for this procedure. Controlled systemic diseases do not contraindicate implant installation, but smoking is a factor that affects implant survival.

Most Frequent Oral Lesions in Patients with Type 2 Diabetes Mellitus.

AIM: Type 2 diabetes mellitus (DM2) is a chronic disease caused by the underproduction of insulin in the organism and it is considered a risk factor to periodontal disease. MATERIALS AND METHODS: This study performed a cross-sectional research on the main oral changes in patients with DM2 and nondiabetics, in Passo Fundo, Rio Grande do Sul, Brazil. The sample included 116 patients examined at the Diabetes Outpatient Clinic of the School of Medicine of the University of Passo Fundo (UPF) and 134 nondiabetic patients examined at the Examinations, Triage, and Emergency Sector of the School of Dentistry of UPF. Inclusion criteria for the study were patients over 35-years old, diagnosed with DM2 for more than 2 years. The same criteria were used for the control group, except for the presence of diabetes. Data collected were analyzed by Statistical Package for the Social Sciences 18.0 for Windows™ software and the Chi-square test at 5% significance. This study showed that, overall, oral lesions were more prevalent in diabetic patients. RESULTS: The stomatological manifestations observed more frequently in such patients were pseudomembranous candidia-sis, lichen planus, lingual varices, xerostomia, and prosthetic stomatitis (p > 0.001). CONCLUSION: Therefore, based on the sample investigated, it is concluded that patients with DM2 present higher prevalence of oral lesions when compared with nondiabetics. CLINICAL SIGNIFICANCE: It is important for the dentist to know about oral lesions because they may allow either early diagnosis in patients unaware of this condition or help diagnosing a potential decompensation. Moreover, oral lesions may represent a potential gateway for infectious agents, and the dentist may restrain this condition by performing treatment as early as possible.

DRB1 locus alleles of HLA class II are associated with modulation of the immune response in different serological profiles of HIV-1/Epstein-Barr virus coinfection in the Brazilian Amazon region.

Background: Epstein-Barr virus (EBV) infection involves distinct clinical and serological profiles. We evaluated the frequency of alleles of locus DRB1 of HLA class II in different serological profiles of EBV infection among HIV-1 infected patients. Methods: We recruited 19 patients with primary infection, 90 with serological transition and 467 with past infection by EBV, HIV-1 co-infection was 100% in primary infection and approximately 70% in other serological profiles. EBV viral load was quantified by real-time PCR, T lymphocyte quantification and cytokine level analysis were performed by flow cytometry, and HLA locus genotyping was performed by PCR-SSO. Results: The DRB1*09 allele was associated with primary infection (p: 0.0477), and carriers of the allele showed changes in EBV viral load (p: 0.0485), CD8(+) T lymphocyte counts (p: 0.0206), double-positive T lymphocyte counts (p: 0.0093), IL-4 levels (p: 0.0464) and TNF levels (p: 0.0161). This allele was also frequent in HIV-coinfected individuals (p: 0.0023) and was related to the log10 HIV viral load (p: 0.0176) and CD8(+) T lymphocyte count (p: 0.0285). In primary infection, the log10 HIV viral load was high (p: 0.0060) and directly proportional to the EBV viral load (p: 0.0412). The DRB1*03 allele correlated with serological transition (p: 0.0477), EBV viral load (p: 0.0015), CD4(+) T lymphocyte count (p: 0.0112), CD8(+) T lymphocyte count (p: 0.0260), double-negative T lymphocyte count (p: 0.0540), IL-4 levels (p: 0.0478) and IL-6 levels (p: 0.0175). In the serological transition group, the log10 HIV viral load was high (p: 0.0060), but it was not associated with the EBV viral load (p: 0.1214). Past infection was related to the DRB1*16 allele (p: 0.0477), with carriers displaying IgG levels (p: 0.0020), CD4(+) T lymphocyte counts (p: 0.0116) and suggestive CD8(+) T count alterations (p: 0.0602). The DRB01*16 allele was also common in HIV-1 patients with past EBV infection (p: 0.0192); however, the allele was not associated with clinical markers of HIV-1 infection. Conclusion: Our results suggest that HLA class II alleles may be associated with the modulation of the serological profiles of the immune response to Epstein-Barr virus infection in patients coinfected with HIV-1.

Glyphosate pollution of surface runoff, stream water, and drinking water resources in Southeast Brazil.

Glyphosate-based herbicides can be harmful to the environment and human health. Especially in developing countries, these herbicides are often used indiscriminately in agricultural and urban areas. Here, we optimized a simple and efficient flow injection-based spectrophotometric method to monitor environmentally relevant glyphosate concentrations in surface waters. The method was then used to assess the environmental mobility of glyphosate in Southeast Brazil by monitoring surface runoff from experimental agricultural soil plots that received glyphosate applications in 2015. Further, water samples from low-order streams were collected in five agricultural, urban, and natural areas, as well as from the 5th-order Rio das Mortes during the rainy season. Finally, 20 drinking water sources were sampled in urban, rural, and agricultural areas. Runoff from reference plots without glyphosate application showed concentrations below the method's detection limit of 0.49 mg.L-1, whereas runoff from plots with standard glyphosate application had concentrations between 1.24 and 6.1 mg.L-1. Similarly, concentrations in natural stream water were below the detection limit, whereas agricultural streams had concentrations of up to 3.7 mg.L-1 (average: 0.97 mg.L-1). In an agricultural stream monitored weekly, concentration peaks were observed after glyphosate applications by farmers, and concentrations were correlated to stream discharge. Urban streams had concentrations of up to 5.8 mg.L-1 (average: 2.6 mg.L-1), but samples from the catchment's major river were mostly below detection limits, illustrating the dilution of urban and agricultural runoff in high-order rivers. In the sampled drinking water resources, glyphosate pollution occurred mainly in the rainy season, with detectable concentrations between 0.5 and 8.7 mg.L-1 in 80% of the sampled drinking water sources. In conclusion, our results suggest considerable environmental mobility of glyphosate in the studied Southeast Brazilian catchment. Substantial pollution, well above national and international limits, was detected in surface runoff, stream water, and drinking water resources.

Development and application of a mini-QuEChERS method for the determination of pesticide residues in anuran adipose tissues.

The expansion of monocultures to regions close to conservation areas has put biodiversity at risk, mainly due to the intense use of pesticides. Anurans are highly susceptible to pesticides and may be a biological marker in the contamination of an area. However, methods for determining pesticides in anurans are incipient. In this work, a miniaturized QuEChERS method was developed for the extraction of atrazine, chlorpyrifos, α- and ß-endosulfan, α-, ß-, θ- and ζ-cypermethrin in anuran adipose tissues. The method was optimized for the tissue sample size scale according to sample mass availability. Extracting solvent and adsorbents for the clean-up step was evaluated, achieving recoveries next to 100% with acetonitrile and without a clean-up step. The mini-QuEChERS method, using 500 mg of adipose tissue, 50 mg of NaCl and 200 mg of MgSO4, 100 µL of ultrapure water, and 1.50 mL of acetonitrile with no purification step, followed by high-performance liquid chromatography analysis and photodiode array detection was validated following the European Community guidelines. The methodology showed a moderate matrix effect for some pesticides, which was corrected using the matrix-matched calibration. The limits of quantification for the pesticide residues in adipose tissues ranged from 10 to 75 µg kg-1. Pesticide recoveries ranged from 74% to 115%, and repeatability and within-lab reproducibility showed relative standard deviations < 11%. The mini-QuEChERS method was applied to extract pesticide residues from the adipose tissues of two species of anurans: Leptodactylus macrosternum and Scinax x-signatus. 25% of samples were positive, detecting endosulfan and chlorpyriphos, confirmed by liquid chromatography coupled to tandem mass spectrometry. The mini-QuEChERS was a simple, economical, and eco-friendly method for extracting pesticide residues in anuran adipose tissue samples.

Characterization of Depression- and Anxiety-Like Behaviours in a Mouse Model of Relapsing-Remitting Multiple Sclerosis.

Relapsing-remitting multiple sclerosis (RRMS) is an autoimmune neurological disease and is the most common subtype of MS. In addition, it is associated with the development of depression and anxiety. To date, depressive- and anxiety-like behaviours were only studied using models of progressive MS, which causes severe motor alterations. Thus, we sought to standardise the depressive and anxiety-like behaviours in an RRMS model induced by experimental autoimmune encephalomyelitis (RR-EAE) in mice. The RR-EAE model was induced in C57BL/6 female mice using myelin oligodendrocyte glycoprotein (MOG35-55) antigen and Quillaja saponin (Quil A) as an adjuvant. The immunisation of RR-EAE did not induce locomotor alteration but caused relapsing-remitting induction of clinical scores in mice until 35 post-immunization (p.i.). Also, increased levels of tumour necrosis factor alpha (TNF-α), astrocyte marker (GFAP), and microglial markers (IBA-1) were detected in the prefrontal cortex at 35 p.i. of RR-EAE. In the open field test, RR-EAE mice showed decreased time spent at the centre and sniffing behaviour (at days 21 and 34 p.i.). Also, on day 35 p.i. the RR-EAE group spent less time in the open arms and had decreased open-arm entries compared to control mice in the elevated plus maze (EPM) test, confirming the anxiety-like behaviour. At day 36° p.i. in the tail suspension test, mice showed depression-like behaviour with decreased latency time and increased immobility time. Thus, the RR-EAE model mimics the neuroinflammatory and behavioural features of the RRMS, including depression- and anxiety-like symptoms.