Cytogenotoxicity evaluation of heavy metals detected in extracts and infusion of Baccharis trimera, potential bioaccumulator plant.

Heavy metals (HMs) are natural components of the Earth's crust that might originate from natural and anthropogenic sources. In excess quantities, the presence of these metals is harmful for both environment and human health. Taking this into account, various investigators examined bioaccumulator species in order to reduce environmental toxicity, among these Baccharis trimera. Therefore, the present study aimed to determine the capacity of B. trimera to bioaccumulate HMs and assess consequent cytogenotoxicity following exposure. B. trimera vegetative parts were collected from two groups (1) control, in which plants were cultivated in soil exposed to distilled water, and (2) exposed, in which plants were cultivated in soil exposed to HMs including manganese (Mn), iron (Fe), lead (Pb), copper (Cu), cobalt (Co), zinc (Zn), and chromium (Cr). HMs were quantified in cultivation soil and extracts (aqueous and ethanolic) as well as infusion of B. trimera vegetative parts. Root lengths and cytogenotoxic effects were determined using Allium cepa test. Results demonstrated that all HMs studied were absorbed and bioaccumulated by B. trimera. Root lengths were decreased when exposed to ethanolic extract of B. trimera cultivated in soil exposed to HMs solution, which was the extract that exhibited the highest cytogenotoxicity values. Thus, data demonstrated that B. trimera might serve as a bioaccumulator for the reduction of environmental toxicity associated with the presence of certain HMs.

Heavy metal accumulation efficiency and subsequent of cytogenotoxicity evaluation in the medicinal plant Equisetum hyemale.

Heavy metals in soils represent a threat to the environment, food safety, as well as human and animal health. The bioaccumulation of these elements in plants might enhance medium- and long-term adverse health risk promoting genetic alterations that lead to dermal, gastrointestinal, circulatory, renal, and brain disorders. The present study aimed to determine the bioaccumulation potential and cytogenotoxic effect of Equisetum hyemale extracts. E. hyemale seedlings were divided into two groups: exposed group (plants cultivated in soil with heavy metals solution) and control (plants cultivated in soil with distilled water). Heavy metals were quantified in the cultivation soils (control and exposed) and extracts (ethanolic and infusion) of vegetative parts from E. hyemale cultivated in both soils. Root length and cytogenotoxic effect were determined utilizing Allium cepa bioassay. Data demonstrated that Equisetum hyemale present the ability to absorb and bioaccumulate different heavy metals including lead, copper, cobalt manganese, zinc, iron and chromium. Given this property E. hyemale may be considered a reliable bioindicator to assess cytogenotoxicity of certain substances that exert adverse risks to environment and human and animal health.

Diagnosis of Schistosoma mansoni infections: what are the choices in Brazilian low-endemic areas?

The population of Brazil is currently characterised by many individuals harbouring low-intensity Schistosoma mansoni infections. The Kato-Katz technique is the diagnostic method recommended by the World Health Organization (WHO) to assess these infections, but this method is not sensitive enough in the context of low egg excretion. In this regard, potential alternatives are being employed to overcome the limits of the Kato-Katz technique. In the present review, we evaluated the performance of parasitological and immunological approaches adopted in Brazilian areas. Currently, the diagnostic choices involve a combination of strategies, including the utilisation of antibody methods to screen individuals and then subsequent confirmation of positive cases by intensive parasitological investigations.

Reduction of Cellular Expression Levels Is a Common Feature of Functionally Affected Pendrin (SLC26A4) Protein Variants.

Sequence alterations in the pendrin gene (SLC26A4) leading to functionally affected protein variants are frequently involved in the pathogenesis of syndromic and nonsyndromic deafness. Considering the high number of SLC26A4 sequence alterations reported to date, discriminating between functionally affected and unaffected pendrin protein variants is essential in contributing to determine the genetic cause of deafness in a given patient. In addition, identifying molecular features common to the functionally affected protein variants can be extremely useful to design future molecule-directed therapeutic approaches. Here we show the functional and molecular characterization of six previously uncharacterized pendrin protein variants found in a cohort of 58 Brazilian deaf patients. Two variants (p.T193I and p.L445W) were undetectable in the plasma membrane, completely retained in the endoplasmic reticulum and showed no transport function; four (p.P142L, p.G149R, p.C282Y and p.Q413R) showed reduced function and significant, although heterogeneous, expression levels in the plasma membrane. Importantly, total expression levels of all of the functionally affected protein variants were significantly reduced with respect to the wild-type and a fully functional variant (p.R776C), regardless of their subcellular localization. Interestingly, reduction of expression may also reduce the transport activity of variants with an intrinsic gain of function (p.Q413R). As reduction of overall cellular abundance was identified as a common molecular feature of pendrin variants with affected function, the identification of strategies to prevent reduction in expression levels may represent a crucial step of potential future therapeutic interventions aimed at restoring the transport activity of dysfunctional pendrin variants.

SARS-CoV-2 breakthrough infections enhance T cell response magnitude, breadth, and epitope repertoire.

Little is known about the effect of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or SARS2) vaccine breakthrough infections (BTIs) on the magnitude and breadth of the T cell repertoire after exposure to different variants. We studied samples from individuals who experienced symptomatic BTIs during Delta or Omicron waves. In the pre-BTI samples, 30% of the donors exhibited substantial immune memory against non-S (spike) SARS2 antigens, consistent with previous undiagnosed asymptomatic SARS2 infections. Following symptomatic BTI, we observed (1) enhanced S-specific CD4 and CD8 T cell responses in donors without previous asymptomatic infection, (2) expansion of CD4 and CD8 T cell responses to non-S targets (M, N, and nsps) independent of SARS2 variant, and (3) generation of novel epitopes recognizing variant-specific mutations. These variant-specific T cell responses accounted for 9%-15% of the total epitope repertoire. Overall, BTIs boost vaccine-induced immune responses by increasing the magnitude and by broadening the repertoire of T cell antigens and epitopes recognized.

Antischistosomal activity of a calcium channel antagonist on schistosomula and adult Schistosoma mansoni worms.

Current schistosomiasis control strategies are largely based on chemotherapeutic agents and a limited number of drugs are available today. Praziquantel (PZQ) is the only drug currently used in schistosomiasis control programs. Unfortunately, this drug shows poor efficacy in patients during the earliest infection phases. The effects of PZQ appear to operate on the voltage-operated Ca2+ channels, which are located on the external Schistosoma mansoni membrane. Because some Ca2+ channels have dihydropyridine drug class (a class that includes nifedipine) sensitivity, an in vitro analysis using a calcium channel antagonist (clinically used for cardiovascular hypertension) was performed to determine the antischistosomal effects of nifedipine on schistosomula and adult worm cultures. Nifedipine demonstrated antischistosomal activity against schistosomula and significantly reduced viability at all of the concentrations used alone or in combination with PZQ. In contrast, PZQ did not show significant efficacy when used alone. Adult worms were also affected by nifedipine after a 24 h incubation and exhibited impaired motility, several lesions on the tegument and intense contractility. These data support the idea of Ca2+ channels subunits as drug targets and favour alternative therapeutic schemes when drug resistance has been reported. In this paper, strong arguments encouraging drug research are presented, with a focus on exploring schistosomal Ca2+ channels.

Schistosoma mansoni in a low-prevalence area in Brazil: the importance of additional methods for the diagnosis of hard-to-detect individual carriers by low-cost immunological assays.

Schistosomiasis diagnosis is based on the detection of eggs in the faeces, which is laborious and lacks sensitivity, especially for patients with a low parasite burden. Immunological assays for specific antibody detection are available, but they usually demonstrate low sensitivity and/or specificity. In this study, two simple immunological assays were evaluated for the detection of soluble Schistosoma mansoni adult worm preparation (SWAP) and egg-specific IgGs. These studies have not yet been evaluated for patients with low parasite burdens. Residents of an endemic area in Brazil donated sera and faecal samples for our study. The patients were initially diagnosed by a rigorous Kato-Katz analysis of 18 thick smears from four different stool samples. The ELISA-SWAP was successful for human diagnosis with 90% sensitivity and specificity, confirming the Kato-Katz diagnosis with nearly perfect agreement, as seen by the Kappa index (0.85). Although the ELISA-soluble S. mansoni egg antigen was 85% sensitive, it exhibited low specificity (80%; Kappa index: 0.75) and was more susceptible to cross-reactivity. We believe that immunological assays should be used in conjunction with Kato-Katz analysis as a supplementary tool for the diagnosis of schistosomiasis for patients with low infection burdens, which are usually hard to detect.

Etiologic and diagnostic evaluation: algorithm for severe to profound sensorineural hearing loss in Brazil.

OBJECTIVE: Evaluation of the effectiveness of imaging and genetic testing, and establishment of a cost-effective diagnostic protocol for the etiologic diagnosis of sensorineural hearing loss (SNHL) in Brazil. DESIGN: Prospective cohort study. STUDY SAMPLE: Analysis of 100 unrelated Brazilian patients with severe to profound bilateral SNHL submitted to cochlear implant (CI) between 2002 and 2010 at the University of Campinas hospital. The study was based upon three groups: individuals with congenital, progressive, and sudden SNHL. RESULTS: After the diagnostic investigation, the number of cases with unknown etiology was reduced from 72 to 42 (a 42% reduction); 25% of cases were due to environmental factors, 19% to genetic causes, and 14% to inner-ear abnormalities or other clinical features. The genetic and imaging findings contributed to the diagnosis of SNHL in 19% and 20% of the cases analysed, respectively. Molecular testing mainly contributed to the diagnosis of patients with congenital SNHL, while the contribution of radiologic examination was higher for individuals with progressive or sudden SNHL. A sequential diagnostic protocol was proposed based on these data. CONCLUSIONS: The proposed diagnostic workup algorithm could provide better optimization of etiologic diagnosis, as well as reduced costs, compared to a simultaneous testing approach.

Prior SARS-CoV-2 Infection Enhances Initial mRNA Vaccine Response with a Lower Impact on Long-Term Immunity.

Spike-encoding mRNA vaccines in early 2021 effectively reduced SARS-CoV-2-associated morbidity and mortality. New booster regimens were introduced due to successive waves of distinct viral variants. Therefore, people now have a diverse immune memory resulting from multiple SARS-CoV-2 Ag exposures, from infection to following vaccination. This level of community-wide immunity can induce immunological protection from SARS-CoV-2; however, questions about the trajectory of the adaptive immune responses and long-term immunity with respect to priming and repeated Ag exposure remain poorly explored. In this study, we examined the trajectory of adaptive immune responses following three doses of monovalent Pfizer BNT162b2 mRNA vaccination in immunologically naive and SARS-CoV-2 preimmune individuals without the occurrence of breakthrough infection. The IgG, B cell, and T cell Spike-specific responses were assessed in human blood samples collected at six time points between a moment before vaccination and up to 6 mo after the third immunization. Overall, the impact of repeated Spike exposures had a lower improvement on T cell frequency and longevity compared with IgG responses. Natural infection shaped the responses following the initial vaccination by significantly increasing neutralizing Abs and specific CD4+ T cell subsets (circulating T follicular helper, effector memory, and Th1-producing cells), but it had a small benefit at long-term immunity. At the end of the three-dose vaccination regimen, both SARS-CoV-2-naive and preimmune individuals had similar immune memory quality and quantity. This study provides insights into the durability of mRNA vaccine-induced immunological memory and the effects of preimmunity on long-term responses.

Immunodiagnostic methods: what is their role in areas of low endemicity?

Worldwide Schistosomiasis mansoni continues to be a serious public health problem. Over the past decades, control programmes have made remarkable progress in reducing S. mansoni infections to a relatively low level in Brazil and African countries. Endemic regions are currently circumscribed in certain core areas where reinfection and repeated chemotherapy are frequent and, consequently, are related to residents with low parasite load. At present, diagnosis is predominately a key step for final disease control although low endemicity area residents are hardly detected by most of the available assays. In this paper, we review the current status and efforts made aiming at the improvement of diagnostic tools for S. mansoni in low endemicity infections. The establishment of diagnostic assays--simple, affordable, sensitive, and specific for field diagnosis of S. mansoni--is essential and should be given high priority.

The effect of waning on antibody levels and memory B cell recall following SARS-CoV-2 infection or vaccination.

As of March 2022, there have been over 450 million reported SARS-CoV-2 cases worldwide, and more than 4 billion people have received their primary series of a COVID-19 vaccine. In order to longitudinally track SARS-CoV-2 antibody levels in people after vaccination or infection, a large-scale COVID-19 sero-surveillance progam entitled SPARTA (SeroPrevalence and Respiratory Tract Assessment) was established early in the pandemic. Anti-RBD antibody levels were tracked in more than 1,000 people. There was no significant decrease in antibody levels during the first 14 months after infection in unvaccinated participants, however, significant waning of antibody levels was observed following vaccination, regardless of previous infection status. Moreover, participants who were pre-immune to SARS-CoV-2 prior to vaccination seroconverted to significantly higher antibody levels, and antibodies were maintained at significantly higher levels than in previously infected, unvaccinated participants. This pattern was entirely due to differences in the magnitude of the initial seroconversion event, and the rate of antibody waning was not significantly different based on the pre-immune status. Participants who received a third (booster) dose of an mRNA vaccine not only increased their anti-RBD antibody levels ∼14-fold, but they also had ∼3 times more anti-RBD antibodies compared to the peak of their antibody levels after receiving their primary vaccine series. In order to ascertain whether the presence of serum antibodies is important for long-term seroprotection, PBMCs from 13 participants who lost all detectable circulating antibodies after vaccination or infection were differentiated into memory cells in vitro . There was a significant recall of memory B cells in the absence of serum antibodies in 70% of the vaccinated participants, but not in any of the infected participants. Therefore, there is a strong connection between anti-RBD antibody levels and the effectiveness of memory B cell recall.

Medicinal plant use in primary health care: an integrative review. / O uso de plantas medicinais na atenção primária à saúde: revisão integrativa.

Medicinal plant (MP) use supports comprehensiveness of care in Primary Health Care (PHC), enabling appreciation of popular knowledge and self-care. This integrative literature review aims to analyze researches that approach the insertion of using MP in PHC. PICO strategy was used as a guideline in search of evidence, reuniting 18 articles published between January 2015 and August 2020, in the Virtual Health Library and PubMed databases. The variables of analysis were knowledge of PHC healthcare professionals about MP and associated policies, MP use by its users, highlighting their profile, the reasons that lead to the use and lack of security in MP use. The results show insufficient knowledge of healthcare professionals about Integrative and Complementary Practices policies and the medicinal uses of plants. The main users are women, elderly, with low income and education, either in Brazil or other countries. Regarding safety in MP use, frequently there is no correct identification of species, its origin, its preparation and the appropriate dose for each case. Finally, failure to approach these contents during training of healthcare professionals generates less knowledge, less research and more prejudice due to lack of information, impairing incentive and dissemination to the community.

O uso de plantas medicinais (PM) favorece a integralidade do cuidado na atenção primária à saúde (APS), valorizando o saber popular e o autocuidado. Esta revisão integrativa de literatura objetiva analisar estudos sobre a inserção do uso de PM na APS. A estratégia PICO norteou a busca de evidências, reunindo 18 artigos publicados de janeiro de 2015 a agosto de 2020 nos bancos de dados Biblioteca Virtual de Saúde e PubMed. As variáveis de análise foram o conhecimento dos profissionais da saúde da APS sobre PM e políticas associadas, o uso de PM pelos usuários, destacando seu perfil, fatores que favorecem o uso e a falta de segurança no uso de PM. A literatura aponta insuficiência de conhecimento dos profissionais de saúde sobre as políticas de práticas integrativas e complementares e o uso de plantas para fins medicinais. Os principais usuários são mulheres, idosas, com baixa renda e escolaridade, tanto no Brasil quanto em outros países. Sobre a segurança no uso de PM, frequentemente não há correta identificação de espécie, origem, preparo e dose adequada para cada caso. Por fim, a não abordagem desses conteúdos durante a formação de profissionais da saúde gera menos conhecimento, menos pesquisas e mais preconceito por falta de informação, prejudicando o incentivo e divulgação à comunidade.

The Effect of Waning on Antibody Levels and Memory B Cell Recall following SARS-CoV-2 Infection or Vaccination.

In order to longitudinally track SARS-CoV-2 antibody levels after vaccination or infection, we assessed anti-RBD antibody levels in over 1000 people and found no significant decrease in antibody levels during the first 14 months after infection in unvaccinated participants, however, a significant waning of antibody levels was observed following vaccination. Participants who were pre-immune to SARS-CoV-2 prior to vaccination seroconverted to higher antibody levels, which were maintained at higher levels than in previously infected, unvaccinated participants. Older participants exhibited lower level of antibodies after vaccination, but a higher level after infection than younger people. The rate of antibody waning was not affected by pre-immunity or age. Participants who received a third dose of an mRNA vaccine not only increased their antibody levels ~14-fold, but also had ~3 times more antibodies compared to when they received their primary vaccine series. PBMC-derived memory B cells from 13 participants who lost all circulating antibodies were differentiated into antibody secreting cells (ASCs). There was a significant recall of memory B cell ASCs in the absence of serum antibodies in 5-8 of the 10 vaccinated participants, but not in any of the 3 infected participants, suggesting a strong connection between antibody levels and the effectiveness of memory B cell recall.

A Comparison of Two Structurally Related Human Milk Oligosaccharide Conjugates in a Model of Diet-Induced Obesity.

Obesity is the largest risk factor for the development of chronic diseases in industrialized countries. Excessive fat accumulation triggers a state of chronic low-grade inflammation to the detriment of numerous organs. To address this problem, our lab has been examining the anti-inflammatory mechanisms of two human milk oligosaccharides (HMOs), lacto-N-fucopentaose III (LNFPIII) and lacto-N-neotetraose (LNnT). LNFPIII and LNnT are HMOs that differ in structure via presence/absence of an α1,3-linked fucose. We utilize LNFPIII and LNnT in conjugate form, where 10-12 molecules of LNFPIII or LNnT are conjugated to a 40 kDa dextran carrier (P3DEX/NTDEX). Previous studies from our lab have shown that LNFPIII conjugates are anti-inflammatory, act on multiple cell types, and are therapeutic in a wide range of murine inflammatory disease models. The α1,3-linked fucose residue on LNFPIII makes it difficult and more expensive to synthesize. Therefore, we asked if LNnT conjugates induced similar therapeutic effects to LNFPIII. Herein, we compare the therapeutic effects of P3DEX and NTDEX in a model of diet-induced obesity (DIO). Male C57BL/6 mice were placed on a high-fat diet for six weeks and then injected twice per week for eight weeks with 25µg of 40 kDa dextran (DEX; vehicle control), P3DEX, or NTDEX. We found that treatment with P3DEX, but not NTDEX, led to reductions in body weight, adipose tissue (AT) weights, and fasting blood glucose levels. Mice treated with P3DEX also demonstrated improvements in glucose homeostasis and insulin tolerance. Treatment with P3DEX or NTDEX also induced different profiles of serum chemokines, cytokines, adipokines, and incretin hormones, with P3DEX notably reducing circulating levels of leptin and resistin. P3DEX also reduced WAT inflammation and hepatic lipid accumulation, whereas NTDEX seemed to worsen these parameters. These results suggest that the small structural difference between P3DEX and NTDEX has significant effects on the conjugates' therapeutic abilities. Future work will focus on identifying the receptors for these conjugates and delineating the mechanisms by which P3DEX and NTDEX exert their effects.

Monoclonal Antibodies Generated against Glycoconjugates Recognize Chemical Linkers.

Monoclonal antibodies (mAbs) that recognize glycans are useful tools to assess carbohydrates' structure and function. We sought to produce IgG mAbs to the human milk oligosaccharide (HMO), lacto-N-fucopentaose III (LNFPIII). LNFPIII contains the Lewisx antigen, which is found on the surface of schistosome parasites. mAbs binding the Lewisx antigen are well-reported in the literature, but mAbs recognizing HMO structures are rare. To generate mAbs, mice were immunized with LNFPIII-DEX (P3DEX) plus CpGs in VacSIM®, a novel vaccine/drug delivery platform. Mice were boosted with LNFPIII-HSA (P3HSA) plus CpGs in Incomplete Freund's Adjuvant (IFA). Splenocytes from immunized mice were used to generate hybridomas and were screened against LNFPIII conjugates via enzyme-linked immunosorbent assay (ELISA). Three positive hybridomas were expanded, and one hybridoma, producing IgG and IgM antibodies, was cloned via flow cytometry. Clone F1P2H4D8D5 was selected because it produced IgG1 mAbs, but rescreening unexpectedly showed binding to both LNFPIII and lacto-N-neotetraose (LNnT) conjugates. To further assess the specificity of the mAb, we screened it on two glycan microarrays and found no significant binding. This finding suggests that the mAb binds to the acetylphenylenediamine (APD) linker-spacer structure of the conjugate. We present the results herein, suggesting that our new mAb could be a useful probe for conjugates using similar linker spacer structures.

Study of modifiers factors associated to mitochondrial mutations in individuals with hearing impairment.

Hearing impairment is the most prevalent sensorial deficit in the general population. Congenital deafness occurs in about 1 in 1000 live births, of which approximately 50% has hereditary cause in development countries. Non-syndromic deafness can be caused by mutations in both nuclear and mitochondrial genes. Mutations in mtDNA have been associated with aminoglycoside-induced and non-syndromic deafness in many families worldwide. However, the nuclear background influences the phenotypic expression of these pathogenic mutations. Indeed, it has been proposed that nuclear modifier genes modulate the phenotypic manifestation of the mitochondrial A1555G mutation in the MTRNR1 gene. The both putative nuclear modifiers genes TRMU and MTO1 encoding a highly conserved mitochondrial related to tRNA modification. It has been hypothesizes that human TRMU and also MTO1 nuclear genes may modulate the phenotypic manifestation of deafness-associated mitochondrial mutations. The aim of this work was to elucidate the contribution of mitochondrial mutations, nuclear modifier genes mutations and aminoglycoside exposure in the deafness phenotype. Our findings suggest that the genetic background of individuals may play an important role in the pathogenesis of deafness-associated with mitochondrial mutation and aminoglycoside-induced.

Serological proteomic screening and evaluation of a recombinant egg antigen for the diagnosis of low-intensity Schistosoma mansoni infections in endemic area in Brazil.

BACKGROUND: Despite decades of use of control programs, schistosomiasis remains a global public health problem. To further reduce prevalence and intensity of infection, or to achieve the goal of elimination in low-endemic areas, there needs to be better diagnostic tools to detect low-intensity infections in low-endemic areas in Brazil. The rationale for development of new diagnostic tools is that the current standard test Kato-Katz (KK) is not sensitive enough to detect low-intensity infections in low-endemic areas. In order to develop new diagnostic tools, we employed a proteomics approach to identify biomarkers associated with schistosome-specific immune responses in hopes of developing sensitive and specific new methods for immunodiagnosis. METHODS AND FINDINGS: Immunoproteomic analyses were performed on egg extracts of Schistosoma mansoni using pooled sera from infected or non-infected individuals from a low-endemic area of Brazil. Cross reactivity with other soil-transmitted helminths (STH) was determined using pooled sera from individuals uniquely infected with different helminths. Using this approach, we identified 23 targets recognized by schistosome acute and chronic sera samples. To identify immunoreactive targets that were likely glycan epitopes, we compared these targets to the immunoreactivity of spots treated with sodium metaperiodate oxidation of egg extract. This treatment yielded 12/23 spots maintaining immunoreactivity, suggesting that they were protein epitopes. From these 12 spots, 11 spots cross-reacted with sera from individuals infected with other STH and 10 spots cross-reacted with the negative control group. Spot number 5 was exclusively immunoreactive with sera from S. mansoni-infected groups in native and deglycosylated conditions and corresponds to Major Egg Antigen (MEA). We expressed MEA as a recombinant protein and showed a similar recognition pattern to that of the native protein via western blot. IgG-ELISA gave a sensitivity of 87.10% and specificity of 89.09% represented by area under the ROC curve of 0.95. IgG-ELISA performed better than the conventional KK (2 slides), identifying 56/64 cases harboring 1-10 eggs per gram of feces that were undiagnosed by KK parasitological technique. CONCLUSIONS: The serological proteome approach was able to identify a new diagnostic candidate. The recombinant egg antigen provided good performance in IgG-ELISA to detect individuals with extreme low-intensity infections (1 egg per gram of feces). Therefore, the IgG-ELISA using this newly identified recombinant MEA can be a useful tool combined with other techniques in low-endemic areas to determine the true prevalence of schistosome infection that is underestimated by the KK method. Further, to overcome the complexity of ELISA in the field, a second generation of antibody-based rapid diagnostic tests (RDT) can be developed.

Interaction between audiology and genetics in the study of a family: the complexity of molecular diagnosis and genetic counseling.

UNLABELLED: Hearing loss is a multifaceted condition with many etiologies, among which genetic mutation is. Therefore, it is important to connect audiological investigation to etiological diagnosis. AIM: this study aims to establish the audiological and genetic profiles of three non-syndromic children with sensorineural hearing loss. MATERIALS AND METHOD: three brothers aged 3, 5 and 16 were enrolled in this study. They were submitted to behavioral and electrophysiological hearing tests and molecular studies. RESULTS: the hearing tests showed moderate to moderately severe bilateral symmetric sensorineural hearing loss and an accentuated descending slope. Transient and Distortion Product Otoacoustic emissions were absent in the two younger children. ABR showed a bilateral moderately severe to severe sensorineural hearing loss. P300 showed bilateral normal latencies in the older brother. Molecular tests showed that the two younger children were heterozygote for mutation 35delG on gene GJB2. CONCLUSION: The combination of speech and hearing tests and genetic analysis allows for the etiologic diagnosis of seemingly similar hearing loss cases, which however display different genetic backgrounds. Molecular studies must be comprehensive enough to avoid precipitated diagnosis which may impair genetic counseling.

Banda ortodôntica como alternativa de tratamento conservador provisório para primeiros molares permanentes com hipomineralização molar incisivo (HMI): relato de caso / Orthodontic banding as an alternative provisional conservative treatment for permanent first molars with incisor molar hypomineralization (IMH): case report

Introdução: a Hipomineralização Molar Incisivo (HMI) é um defeito qualitativo de desenvolvimento de esmalte que pode ocasionar fraturas pós-eruptivas (FPE), lesões de cárie e sensibilidade. Objetivo: relatar o tratamento de HMI severa através da cimentação de bandas ortodônticas para preservação da estrutura dentária em primeiros molares permanentes inferiores com FPE. Relato do caso: criança do sexo feminino, 10 anos de idade, apresentou-se com queixa de hipersensibilidade e fratura dentária associada à restauração prévia. Clinicamente, observou-se presença de HMI severa, com FPE associada à lesão de cárie em dentina nas superfícies oclusal e vestibular do dente 36 e restauração insatisfatória com cimento de ionômero de vidro (CIV) na superfície vestibular do dente 46 que apresentava opacidades demarcadas branco-creme. Radiograficamente, observou-se ausência de comprometimento pulpar. Após manejo por meio de abordagens não-invasivas (controle de biofilme e dieta e aplicação de verniz fluoretado), o tratamento proposto foi a cimentação de banda ortodôntica com CIV modificado por resina (Riva Light Cure®, SDI) nos dentes 36 e 46 para maior longevidade das restaurações. O tratamento restaurador atraumático (TRA) foi realizado no dente 36 previamente à cimentação da banda ortodôntica. Resultados: após o tratamento, a criança não relatou dor ou desconforto e as restaurações mantiveram-se intactas. A mãe da criança foi orientada quanto à importância de acompanhamento periódico a cada 4 meses. Conclusão: a cimentação das bandas ortodônticas com CIV possibilitou o manejo conservador de molares permanentes com HMI severa, com manutenção de sua funcionalidade oclusal, saúde pulpar e gengival, proporcionando melhor qualidade de vida à paciente.

Introduction: molar Incisor Hypomineralization (MIH) is a qualitative developmental enamel defect that can cause posteruptive enamel breakdown (PEB), caries lesions, and sensitivity. Objective: to report the treatment of a child with severe MIH through the cementation of orthodontic bands in lower first permanent molars with PEB to preserve tooth structure. Case report: female child, 10 years old, presenting hypersensitivity complaints and tooth fracture associated with previous restoration. Severe MIH was observed, with PEB associated with dentin caries on the occlusal and buccal surfaces of tooth #36 and unsatisfactory glass ionomer cement (GIC) restoration on the buccal surface of tooth #46 which had creamy-white marked opacities. There was no pulp involvement radiographically. After management through non-invasive approaches (biofilm and diet control and application of fluoride varnish), the proposed treatment was the cementation of an orthodontic band with resin-modified GIC (Riva Light Cure®, SDI) on teeth #36 and #46 to long-term lifespan restorations. Atraumatic restorative treatment (ART) was performed on tooth #36 prior to the cementation of the orthodontic band. Results: after treatment, the child did not report pain or discomfort and the restorations remained intact. The child's mother was instructed about the importance of periodic follow-up visits every 4 months. Conclusion: the cementation of orthodontic bands with GIC allowed the conservative management of permanent molars with severe MIH, maintaining their functional occlusion, pulpal and gingival health, providing a better quality of life to the patient.

Implantação do Time de Resposta Rápida em um Hospital Público Pediátrico

Este artigo tem por objetivo descrever a experiência na implantação e condução do trabalho do Time Reposta Rápida em um hospital público pediátrico 100% Sistema Único de Saúde, indicando fatores que funcionaram como facilitadores e dificultadores desta implantação. Realizado estudo descritivo, qualitativo, do tipo relato de experiência. Na implantação do Time Reposta Rápida na enfermaria A do Hospital Infantil Nossa Senhora da Glória (Milena Gottardi), foram identificados vários fatores que funcionaram como facilitadores para esta implantação, dentre eles o envolvimento de grande parte da equipe em todas as etapas. Quanto aos dificultadores cita-se a demora na aquisição dos equipamentos, materiais e insumos, face às questões legais e burocráticas no processo de licitação e compras; grande esforço para adaptar as escalas dos profissionais a realidade do serviço; demora no processo de capacitação dos médicos da equipe; resistência de alguns profissionais por não compreenderem a magnitude do Time Reposta Rápida na realidade local. A implantação do Time Reposta Rápida pode oferecer subsídios para reorganização dos processos de trabalho.