Neonatal screening for spinal muscular atrophy: A pilot study in Brazil.

Spinal muscular atrophy (SMA) is considered one of the most common autosomal recessive disorders, with an estimated incidence of 1 in 10,000 live births. Testing for SMA has been recommended for inclusion in neonatal screening (NBS) panels since there are several therapies available and there is evidence of greater efficacy when introduced in the pre/early symptomatic phases. In Brazil, the National Neonatal Screening Program tests for six diseases, with a new law issued in 2021 stating that it should incorporate more diseases, including SMA. In the present study, dried blood spot (DBS) samples collected by the Reference Services of Neonatal Screening of RS and SP, to perform the conventional test were also screened for SMA, using real-time PCR, with SALSA MC002 technique. A total of 40,000 samples were analyzed, enabling the identification of four positive cases of SMA, that were confirmed by MLPA. Considering our sampling, Brazil seems to have an incidence comparable to the described in other regions. This work demonstrated that the use of the MC002 technique in samples routinely collected for the conventional NBS program is suitable to screen for SMA in our conditions and can be included in the expansion of the neonatal screening programs.

The impact of neonatal 17-hydroxyprogesterone cutoff determination in a public newborn screening program for congenital adrenal hyperplasia in Southern Brazil: 3 years' experience.

Congenital adrenal hyperplasia (CAH) occurs due to enzyme defects in adrenal steroidogenesis. The 21-hydroxylase deficiency accounts for 90-95% of cases, triggering accumulation of 17-hydroxyprogesterone (17-OHP). Early diagnosis through neonatal screening allows adequate treatment and reduced mortality. The purpose of the study was to determine 17-OHP cutoffs for the diagnosis of CAH in a public newborn screening program in Southern Brazil. A retrospective, descriptive, cross-sectional study was conducted to analyze 17-OHP levels in dried blood samples collected on filter paper of 317,745 newborns screened at a public newborn screening center from May 2014 to April 2017. Neonatal 17-OHP was measured in DBS samples using a time-resolved fluoroimmunoassay (GSP® kit 3305-0010; PerkinElmer). Different cutoffs were determined and stratified by birth weight. The incidence of CAH was 1:15,887 live births in the state of Rio Grande do Sul, with 20 cases of classical CAH diagnosed during the study period. Most newborns (80.73%) were white, and the prematurity rate was 9.8% in the study population. The combination of different percentiles, 98.5th for birth weight 2001-2500 g and 99.8th for the other birth weight groups, decreased false-positive results and increased specificity compared with current reference values to identify classical CAH cases. The local 17-OHP cutoffs determined were higher than those currently used by this screening program for all birth weight groups. The calculation of reference values from local population data and the combination of percentiles proved to be a valuable tool for proper diagnosis of CAH and reduction in the number of false positives.

Heterogeneity of variance and genetic parameters for milk production in cattle, using Bayesian inference.

The goal of this study was to verify the effect of heterogeneity of variance (HV) on milk production in up to 305 days of lactation (L305) of daughters of Girolando, Gir and Holstein sires, as well as in the genetic evaluation of these sires and their progenies. in Brazil. The model included contemporary groups (consisting of herd, year and calving season) as a fixed effect, cow age at calving (linear and quadratic effects) and heterozygosity (linear effect) as covariates, in addition to the random effects of direct additive genetic and environmental, permanent and residual. The first analysis consisted of the single-trait animal model, with L305 records (disregarding HV). The second considered classes of standard deviations (SD): two-trait model including low and high classes (considering HV), according to the standardized means of L305 for herd-year of calving. The low SD class was composed of herds with SD equal to or less than zero and the high class with positive SD values. Estimates of (co)variance components and breeding values were obtained separately for each scenario using Bayesian inference via Gibbs sampling. Different heritability was estimated. Higher for the high DP class in the Gir (0.20) and Holstein (0.15) breeds, not occurring the same in the Girolando breed, with a lower value among the classes for the high DP (0.10). High values of genetic correlations were also found between low and high SD classes (0.88; 0.85 and 0.79) for the Girolando, Gir and Holstein breeds, respectively. Like the order correlations (Spearman) which were also high for the three breeds analyzed (equal to or above 0.92). Thus, the presence of HV had a smaller impact for L305 and did not affect the genetic evaluation of sires.

Epidemiological profile of congenital hypothyroidism at a southern Brazilian state.

Objective: To determine the incidence of congenital hypothyroidism (CH) over a 10-year period at the Reference Service in Neonatal Screening of the state of Rio Grande do Sul (RSNS-RS). Subjects and methods: Historical cohort study including all newborns screened for CH by the RSNS-RS from January 2008 until December 2017. Data of all newborns with neonatal TSH (neoTSH; heel prick test) values ≥ 9 mIU/L were collected. According to neoTSH values, the newborns were allocated into two groups: Group 1 (G1), comprising newborns with neoTSH ≥ 9 mIU/L and serum TSH (sTSH) < 10 mIU/L, and Group 2 (G2), comprising those with neoTSH ≥ 9 mIU/L and sTSH ≥ 10 mIU/L. Results: Of 1,043,565 newborns screened, 829 (0.08%) had neoTSH values ≥ 9 mIU/L. Of these, 284 (39.3%) had sTSH values < 10 mIU/L and were allocated to the G1 group, while 439 (60.7%) had sTSH ≥ 10 mIU/L and were allocated to the G2 group, and 106 (12.7%) were considered missing data. The overall incidence of CH was 42.1 per 100,000 newborns screened (95% confidence interval [CI] 38.5-45.7/100,000) or 1:2377 screened newborns. The sensibility and specificity of neoTSH ≥ 9 mIU/L were 97% and 11%; of neoTSH 12.6 mUI/L, 73% and 85% respectively. Conclusion: In this population, the incidence of permanent and transitory CH was 1:2377 screened newborns. The neoTSH cutoff value adopted during the study period showed excellent sensibility, which matters for a screening test.

Perspectives of economic losses due to condemnation of cattle and buffalo carcasses in the northern region of Brazil.

The work aims to study the economical losses of the condemnation of bovine and buffalo carcasses, in order to estimate the losses in animals slaughtered in Santarém-Pará, Brazil, between 2016 and 2018, with data obtained from the Municipal Department of Agriculture and Fisheries. Sex, age, origin, total number of animals slaughtered and causes of condemnation of carcasses were considered. All analyzes were performed in RStudio version 1.1.463. In this study, 71,277 bovine carcasses and 2,016 buffalo carcasses were inspected, of which 300 bovine and 71 buffalo were condemned. The highest prevalence of causes of condemnation in cattle was recorded for brucellosis (0.0020%) and tuberculosis (0.0019%). In buffaloes, tuberculosis (0.0307%) peritonitis (0,0019%) were the main causes of condemnations. Economical losses were more evident in females, for both species. The projection of economical losses related to the condemnation of carcasses showed a sharp growth for the next three years, if the average growth remains constant. The biggest projected loss was for bovine females, with an accumulated projection of $ 5,451.44. The smallest estimated loss was for buffalo males, projected at more than thirty-two thousand reais. The most important causes of condemnation report the diseases brucellosis and tuberculosis, as the ones with the greatest impact. In the buffalo species this was even more accentuated, even though the number of buffaloes slaughtered is more than 35 times smaller than the number of cattle.

Evaluation of RET-He values as an early indicator of iron deficiency anemia in pregnant women

Abstract Introduction During pregnancy, women are at an increased risk of developing iron-deficiency anemia. Objective The objective of this study was to assess the diagnostic performance of the reticulocyte hemoglobin equivalent (RET-He) in the early detection of iron-deficiency anemia in a group of pregnant women and to establish a reference range for this parameter in a group of control individuals. Method: A total of 60 patients and 130 control subjects were included in the study. Blood samples collected from the subjects were submitted to a complete blood count and a serum ferritin test and the data were analyzed by comparing the groups and ROC curves. Results The reference range found for the RET-He was between 29.75pg and 38.24pg, with a median of 35pg. The receiver operating characteristic (ROC) curve analysis for the ferritin parameter showed an area under the curve of 0.732 for the RET-He, 0.586 for hemoglobin, 0.551 for the mean corpuscular hemoglobin concentration and 0.482 for the mean corpuscular volume. Conclusion Early diagnosis of iron deficiency anemia in pregnancy is essential to prevent damage to both maternal and fetal health. The RET-He presents an excellent potential as an auxiliary tool for the diagnosis of iron deficiency in pregnant women.

Evaluation of RET-He values as an early indicator of iron deficiency anemia in pregnant women.

INTRODUCTION: During pregnancy, women are at an increased risk of developing iron-deficiency anemia. OBJECTIVE: The objective of this study was to assess the diagnostic performance of the reticulocyte hemoglobin equivalent (RET-He) in the early detection of iron-deficiency anemia in a group of pregnant women and to establish a reference range for this parameter in a group of control individuals. METHOD: A total of 60 patients and 130 control subjects were included in the study. Blood samples collected from the subjects were submitted to a complete blood count and a serum ferritin test and the data were analyzed by comparing the groups and ROC curves. RESULTS: The reference range found for the RET-He was between 29.75pg and 38.24pg, with a median of 35pg. The receiver operating characteristic (ROC) curve analysis for the ferritin parameter showed an area under the curve of 0.732 for the RET-He, 0.586 for hemoglobin, 0.551 for the mean corpuscular hemoglobin concentration and 0.482 for the mean corpuscular volume. CONCLUSION: Early diagnosis of iron deficiency anemia in pregnancy is essential to prevent damage to both maternal and fetal health. The RET-He presents an excellent potential as an auxiliary tool for the diagnosis of iron deficiency in pregnant women.

Epidemiological profile of congenital hypothyroidism at a southern Brazilian state

ABSTRACT Objective: To determine the incidence of congenital hypothyroidism (CH) over a 10-year period at the Reference Service in Neonatal Screening of the state of Rio Grande do Sul (RSNS-RS). Subjects and methods: Historical cohort study including all newborns screened for CH by the RSNS-RS from January 2008 until December 2017. Data of all newborns with neonatal TSH (neoTSH; heel prick test) values ≥ 9 mIU/L were collected. According to neoTSH values, the newborns were allocated into two groups: Group 1 (G1), comprising newborns with neoTSH ≥ 9 mIU/L and serum TSH (sTSH) < 10 mIU/L, and Group 2 (G2), comprising those with neoTSH ≥ 9 mIU/L and sTSH ≥ 10 mIU/L. Results: Of 1,043,565 newborns screened, 829 (0.08%) had neoTSH values ≥ 9 mIU/L. Of these, 284 (39.3%) had sTSH values < 10 mIU/L and were allocated to the G1 group, while 439 (60.7%) had sTSH ≥ 10 mIU/L and were allocated to the G2 group, and 106 (12.7%) were considered missing data. The overall incidence of CH was 42.1 per 100,000 newborns screened (95% confidence interval [CI] 38.5-45.7/100,000) or 1:2377 screened newborns. The sensibility and specificity of neoTSH ≥ 9 mIU/L were 97% and 11%; of neoTSH 12.6 mUI/L, 73% and 85% respectively. Conclusion: In this population, the incidence of permanent and transitory CH was 1:2377 screened newborns. The neoTSH cutoff value adopted during the study period showed excellent sensibility, which matters for a screening test.

Escuta do Sofrimento Mental Estudantil: Relato de Experiência do Atendimento Psicológico aos Estudantes da UFRJ / Listening to Student Mental Suffering: Experience Report of Psychological Care to UFRJ Students / Escuchando el sufrimiento Mental de los Estudiantes: Experiencia Informe de Atención Psicológica de la UFRJ

Relato de experiência acadêmica de três estagiários de graduação em Psicologia da Universidade Federal do Rio de Janeiro (UFRJ) no atendimento psicológico para estudantes da universidade. O objetivo é destacar as principais questões de sofrimento psíquico relatadas pelos estudantes atendidos e elucidar a importância da escuta clínica no campo acadêmico. Analisaram-se os aspectos recorrentes emergidos durante os atendimentos e presentes na discussão dos casos durante as supervisões com a professora orientadora. Do total de 24 estudantes atendidos, as principais queixas sintomatológicas dos participantes foram crises de ansiedade, sintomas de depressão e de ideação suicida. Buscou-se realizar um paralelo entre as dificuldades acadêmicas e emocionais com a implicação subjetiva dos estudantes na instituição UFRJ. Além disso, a questão do suicídio também foi destacada como o maior desafio clínico pelos estagiários nos atendimentos. Ao final verificou-se a importância do atendimento psicológico para os estudantes dentro do espaço acadêmico, pois esse lugar também de escuta é uma forma de acolhimento integral do estudante.

Report of academic experience from a team of graduate students in Psychology at the Federal University of Rio de Janeiro (UFRJ) providing psychological assistance to other students at this university. The objective is to highlight the main issues of psychological suffering reported by the students assisted and to elucidate the importance of clinical listening in the academic field. The recurrent aspects that emerged during the consultations and were present in the discussion of cases during the supervisions with the mentor teacher were analysed. Of the total of 24 students attended, the main symptomatological complaints of the participants were anxiety crises, symptoms of depression and suicidal ideation. We sought to make a parallel between academic and emotional difficulties, and the subjective implication of UFRJ students. In addition, the issue of suicide was also highlighted as the greatest clinical challenge by trainees in care. It was found that psychological assistance for students within the academic space is of huge importance, for the provision of a careful listening is also a kind of integral reception.

Informe de experiencia académica de tres pasantes de licenciatura en Psicología en la Universidad Federal de Río de Janeiro (UFRJ) en atención psicológica a estudiantes universitarios. El objetivo es resaltar los principales problemas del sufrimiento psicológico reportados por los estudiantes atendidos y dilucidar la importancia de la escucha clínica en el ámbito académico. Se analizaron aspectos recurrentes que surgieron durante las consultas y presentes en las discusiones de casos durante las supervisiones con el profesor guía. Del total de 24 estudiantes atendidos, las principales quejas sintomáticas de los participantes fueron crisis de ansiedad, síntomas de depresión e ideación suicida. Se buscó establecer un paralelismo entre las dificultades académicas y emocionales con la participación subjetiva de los estudiantes en UFRJ. Además, también se destacó el tema del suicidio como el mayor desafío clínico al que se enfrentan los internos en las consultas. En conclusión, se verificó la importancia del cuidado psicológico para los estudiantes dentro del espacio académico, ya que este lugar de escucha es también una forma de acogida integral para el estudiante.

Chemically enhanced primary treatment of dairy wastewater using chitosan obtained from shrimp wastes: optimization using a Doehlert matrix design.

Dairy operations generate large volumes of polluted wastewater that require treatment prior to discharge. Chemically enhanced primary treatment (CEPT) is a widely utilized wastewater treatment strategy; but it requires the use of non-biodegradable coagulants that can lead to toxic-byproducts. In this study, chitin from shrimp shell waste is extracted and converted into chitosan. Chitosan was demonstrated to be a natural, low-cost alternative coagulant compatible with the CEPT. Following treatment, dissolved air flotation allowed for the removal of turbidity, COD, and UV254 from the synthetic dairy effluent (SDE). Doehlert matrix was used to optimize the chitosan dosage and pH of the CEPT; as well as to model the process. The mechanisms behind the coagulation-flocculation were revealed using zeta potential analysis. FTIR spectroscopy was utilized to confirm the functional groups present on the chitosan. Chitosan with a degree of deacetylation equal to 81% was obtained. A chitosan dose of 73.34 mg/L at pH 5.00 was found to be optimal for the removal of pollutants. Removals of COD, turbidity and UV254 were 77.5%, 97.6%, and 88.8%, respectively. The amount of dry sludge generated to treat 1 m³ of SDE was 0.041 kg. Coagulation-flocculation mechanisms involved in chitosan-mediated treatment of SDE involve the neutralization of electrostatic charges carried on the amine groups present in cationic chitosan at pH 5.00. Doehlert matrix proved to be a useful tool in optimizing parameters throughout the coagulation-flocculation process. Chitosan from shrimp waste is a low-cost, eco-friendly coagulant alternative for the removal pollutants from dairy effluent using the CEPT.

A pre-formulation study of tetracaine loaded in optimized nanostructured lipid carriers.

Tetracaine (TTC) is a local anesthetic broadly used for topical and spinal blockade, despite its systemic toxicity. Encapsulation in nanostructured lipid carriers (NLC) may prolong TTC delivery at the site of injection, reducing such toxicity. This work reports the development of NLC loading 4% TTC. Structural properties and encapsulation efficiency (%EE > 63%) guided the selection of three pre-formulations of different lipid composition, through a 23 factorial design of experiments (DOE). DLS and TEM analyses revealed average sizes (193-220 nm), polydispersity (< 0.2), zeta potential |- 21.8 to - 30.1 mV| and spherical shape of the nanoparticles, while FTIR-ATR, NTA, DSC, XRD and SANS provided details on their structure and physicochemical stability over time. Interestingly, one optimized pre-formulation (CP-TRANS/TTC) showed phase-separation after 4 months, as predicted by Raman imaging that detected lack of miscibility between its solid (cetyl palmitate) and liquid (Transcutol) lipids. SANS analyses identified lamellar arrangements inside such nanoparticles, the thickness of the lamellae been decreased by TTC. As a result of this combined approach (DOE and biophysical techniques) two optimized pre-formulations were rationally selected, both with great potential as drug delivery systems, extending the release of the anesthetic (> 48 h) and reducing TTC cytotoxicity against Balb/c 3T3 cells.

Mucoadhesive, Thermoreversible Hydrogel, Containing Tetracaine-Loaded Nanostructured Lipid Carriers for Topical, Intranasal Needle-Free Anesthesia.

Recent advances have been reported for needle-free local anesthesia in maxillary teeth by administering a nasal spray of tetracaine (TTC) and oxymetazoline, without causing pain, fear, and stress. This work aimed to assess whether a TTC-loaded hybrid system could reduce cytotoxicity, promote sustained permeation, and increase the anesthetic efficacy of TTC for safe, effective, painless, and prolonged analgesia of the maxillary teeth in dental procedures. The hybrid system based on TTC (4%) encapsulated in nanostructured lipid carriers (NLC) and incorporated into a thermoreversible hydrogel of poloxamer 407 (TTCNLC-HG4%) displayed desirable rheological, mechanical, and mucoadhesive properties for topical application in the nasal cavity. Compared to control formulations, the use of TTCNLC-HG4% slowed in vitro permeation of the anesthetic across the nasal mucosa, maintained cytotoxicity against neuroblastoma cells, and provided a three-fold increase in analgesia duration, as observed using the tail-flick test in mice. The results obtained here open up perspectives for future clinical evaluation of the thermoreversible hybrid hydrogel, which contains TTC-loaded NLC, with the aim of creating an effective, topical, intranasal, needle-free anesthesia for use in dentistry.

Rhesus macaques self-curing from a schistosome infection can display complete immunity to challenge.

The rhesus macaque provides a unique model of acquired immunity against schistosomes, which afflict >200 million people worldwide. By monitoring bloodstream levels of parasite-gut-derived antigen, we show that from week 10 onwards an established infection with Schistosoma mansoni is cleared in an exponential manner, eliciting resistance to reinfection. Secondary challenge at week 42 demonstrates that protection is strong in all animals and complete in some. Antibody profiles suggest that antigens mediating protection are the released products of developing schistosomula. In culture they are killed by addition of rhesus plasma, collected from week 8 post-infection onwards, and even more efficiently with post-challenge plasma. Furthermore, cultured schistosomula lose chromatin activating marks at the transcription start site of genes related to worm development and show decreased expression of genes related to lysosomes and lytic vacuoles involved with autophagy. Overall, our results indicate that enhanced antibody responses against the challenge migrating larvae mediate the naturally acquired protective immunity and will inform the route to an effective vaccine.

Definition of Reference Range for the Immature Granulocytes Parameter Provided by a Hematology Analyzer.

BACKGROUND: Some pathologies or physiological changes may show forms of granulocytes in peripheral blood. Hematology analyzers have brought new parameters such as the detection of immature granulocytes (IG), which may be useful biomarkers. The objective of this study was determined the IG count in a control group to establish the reference range. METHODS: A group of healthy donors was used to obtain the reference value of IG in the laboratory of the Hospital de Clínicas de Porto Alegre. RESULTS: The reference range of IG (n = 115) was 0 - 0.06 x 109/L and 0 - 0.63%. This reference interval was similar to that of previous studies. CONCLUSIONS: The determination of the specific reference interval for each region is an important tool in the direct application of biomarkers in clinical laboratory routines. The use of reference values is essential so that the true positive cases for microscopic review are detected without a large number of false positives, which would impair laboratory efficiency.

Twenty Years of Neonatal Screening for Sickle Cell Disease in Brazil: The Challenges of a Continental Country with High Genetic Heterogeneity

Abstract Sickle cell disease (SCD) is the most common inherited hematological disease worldwide. The benefits of diagnosis and early intervention have led to the wide dissemination of public health programs worldwide. Through neonatal screening programs, it is possible to reduce morbidity and mortality in the first 5 years of life. The prophylactic use of penicillin, the anti-pneumococcal vaccine and other intensive care, increase the survival and quality of life of people with SCD. The aim of this study is to present the 20-year history of screening for hemoglobinopathies in Brazil and its challenges. From 2001 to 2019, an average of 2,400,000 children were screened per year nationwide, with the coverage being of 82,16%. The screening of 54,9% of newborns is collected up to their 5th day of life. The prevalence of SCD was 1:2,263 newborns; therefore, it was the second most-common disease detected by the program of Brazil, being only after hypothyroidism (1/2,175 live births). The healthcare system should provide the necessary infrastructure to confirm the diagnosis of newborns and to provide appropriate counseling and treatment. The early diagnosis and treatment, as well as the follow-up with a multidisciplinary team, are fundamental to the survival rate and the quality of life of patients.

Rhesus macaques self-curing from a schistosome infection can display complete immunity to challenge

The rhesus macaque provides a unique model of acquired immunity against schistosomes, which afflict >200 million people worldwide. By monitoring bloodstream levels of parasite-gut-derived antigen, we show that from week 10 onwards an established infection with Schistosoma mansoni is cleared in an exponential manner, eliciting resistance to reinfection. Secondary challenge at week 42 demonstrates that protection is strong in all animals and complete in some. Antibody profiles suggest that antigens mediating protection are the released products of developing schistosomula. In culture they are killed by addition of rhesus plasma, collected from week 8 post-infection onwards, and even more efficiently with post-challenge plasma. Furthermore, cultured schistosomula lose chromatin activating marks at the transcription start site of genes related to worm development and show decreased expression of genes related to lysosomes and lytic vacuoles involved with autophagy. Overall, our results indicate that enhanced antibody responses against the challenge migrating larvae mediate the naturally acquired protective immunity and will inform the route to an effective vaccine.

Functional Hybrid Nanoemulsions for Sumatriptan Intranasal Delivery.

In recent years, advanced nanohybrid materials processed as pharmaceuticals have proved to be very advantageous. Triptans, such as the commercially available intranasal sumatriptan (SMT), are drugs employed in the treatment of painful migraine symptoms. However, SMT effectiveness by the intranasal route is limited by its high hydrophilicity and poor mucoadhesion. Therefore, we designed hybrid nanoemulsions (NE) composed of copaiba oil as the organic component plus biopolymers (xanthan, pectin, alginate) solubilized in the continuous aqueous phase, aiming at the intranasal release of SMT (2% w/v). Firstly, drug-biopolymer complexes were optimized in order to decrease the hydrophilicity of SMT. The resultant complexes were further encapsulated in copaiba oil-based nanoparticles, forming NE formulations. Characterization by FTIR-ATR, DSC, and TEM techniques exposed details of the molecular arrangement of the hybrid systems. Long-term stability of the hybrid NE at 25°C was confirmed over a year, regarding size (~ 120 nm), polydispersity (~ 0.2), zeta potential (~ -25 mV), and nanoparticle concentration (~ 2.1014 particles/mL). SMT encapsulation efficiency in the formulations ranged between 41-69%, extending the in vitro release time of SMT from 5 h (free drug) to more than 24 h. The alginate-based NE was selected as the most desirable system and its in vivo nanotoxicity was evaluated in a zebrafish model. Hybrid NE treatment did not affect spontaneous movement or induce morphological changes in zebrafish larvae, and there was no evidence of mortality or cardiotoxicity after 48 h of treatment. With these results, we propose alginate-based nanoemulsions as a potential treatment for migraine pain.

Hybrid nanofilms as topical anesthetics for pain-free procedures in dentistry.

Topical anesthetics are widely applied in order to relieve the discomfort and anxiety caused by needle insertion and other painful superficial interventions at the oral cavity. So far, there are no commercially available effective topical anesthetic formulations for that purpose, and the most of developments are related to hydrophilic and low mucoadhesive forms. Therefore, we have prepared different hybrid nanofilms composed of biopolymer matrices (chitosan, pectin, and chitosan-pectin) blended with nanostructured lipid carriers (NLC) loading the eutectic mixture of 5% lidocaine-prilocaine (LDC-PLC), in order to fulfill this gap in the market. These dual systems were processed as hybrid nanofilms by the solvent/casting method, and its mucoadhesive, structural and mechanical properties were detailed. The most appropriate hybrid nanofilm combined the advantages of both pectin (PCT) and NLC components. The resultant material presented sustained LDC-PLC release profile for more than 8 h; permeation across porcine buccal mucosa almost twice higher than control and non-cytotoxicity against 3T3 and HACAT cell lines. Then, the in vivo efficacy of PCT/NLC formulation was compared to biopolymer film and commercial drug, exhibiting the longest-lasting anesthetic effect (> 7 h), assessed by tail flick test in mice. These pectin-based hybrid nanofilms open perspectives for clinical trials and applications beyond Dentistry.

Cystic Fibrosis: A Simple and Customized Strategy for Genetic Screening Able to Detect Over 90% of Identified Mutated Alleles in Brazilian Newborns.

INTRODUCTION: The incorporation of molecular genetic testing into cystic fibrosis (CF) screening programs increases the specificity of the diagnostic strategy and has the potential to decrease the rate of false- positive results. In this sense, our objective was to develop a genotyping assay that could detect 25 pathogenic variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene with high sensitivity and that could be incorporated into the routine of newborn screening, complementing the current existing protocol used in our public health institution. METHODS: A mini-sequencing assay was standardized using single-base extension in a previously genotyped control sample. This strategy was validated in a Brazilian cohort of CF patients by Sanger sequencing. RESULTS: The inclusion of the 25 variants in the current newborn screening program increased the identification rates of two alleles from 33 to 52.43% in CF patients. This new approach was able to detect a total of 37 variants, which represents 93.01% of all mutated alleles described in the last CF Brazilian Register. CONCLUSIONS: Mini-sequencing for the simultaneous detection of 25 CFTR gene variants improves the screening of Brazilian newborns and decreases the number of inconclusive cases. This method uses minimal hands-on time and is suited for rapid screening, which reduces sample processing costs.

Improved efficacy of naproxen-loaded NLC for temporomandibular joint administration.

Inflammatory conditions of the temporomandibular joint (TMJ) and peripheral tissues affect many people around the world and are commonly treated with non-steroidal anti-inflammatory drugs (NSAIDs). However, in order to get desirable results, treatments with NSAIDs may take weeks, causing undesirable side effects and requiring repeated administration. In this sense, this work describes the development of an optimized nanostructured lipid carrier (NLC) formulation for intra-articular administration of naproxen (NPX). An experimental design (23) selected the best formulation in terms of its physicochemical and structural properties, elucidated by different methods (DLS, NTA, TEM, DSC, and ATR-FTIR). The chosen formulation (NLC-NPX) was tested on acute inflammatory TMJ nociception, in a rat model. The optimized excipients composition provided higher NPX encapsulation efficiency (99.8%) and the nanoparticles were found stable during 1 year of storage at 25 °C. In vivo results demonstrated that the sustained delivery of NPX directly in the TMJ significantly reduced leukocytes migration and levels of pro-inflammatory cytokines (IL-1ß and TNF-α), for more than a week. These results point out the NLC-NPX formulation as a promising candidate for the safe treatment of inflammatory pain conditions of TMJ or other joints.