Corrigendum: Evaluation of PIK3CA mutations in advanced ER+/HER2-breast cancer in Portugal - U-PIK Project.

[This corrects the article DOI: 10.3389/fmolb.2023.1082915.].

AdultCarbQuiz for the Portuguese population with type 1 diabetes mellitus: translation, cultural adaptation and validation of its metabolic impact.

AIMS: In type 1 diabetes mellitus (T1DM), functional insulin therapy, based on carbohydrate (CH) counting and individualized insulin-to-carbohydrate ratio, is essential to achieve an adequate metabolic control. However, to date, few tools have been validated to assess patients' knowledge about CH counting, with the AdultCarbQuiz questionnaire having proved to be a reliable method in an American cohort. The aim of this study was to translate, culturally adapt and validate the AdultCarbQuiz questionnaire for the Portuguese population.  METHODS: This was a cross-sectional study of patients with T1DM on functional insulin therapy through continuous subcutaneous insulin infusion (CSII). Prior to its application, the AdultCarbQuiz questionnaire was translated and culturally adapted to the Portuguese context. Statistical analyses include descriptive, correlation and intern consistency analysis using IBM® SPSS® Statistics, version 27. RESULT: One hundred patients were included, 58% of female sex, with a mean age of 31.09 ± 10.77 years. Mean disease duration was 15.04 ± 9.23 years, and mean CSII usage time was 4.02 ± 3.90 years. The average value of glycated haemoglobin (HbA1c), time in range (TIR), time above range (TAR) and time below range was, respectively, 7.32 ± 0.87, 59.75 ± 14.13, 34.38 ± 15.40 and 5.75 ± 6.58%. The average score of the questionnaire was 30.86 points ± 3.58 points, considered high. The Kuder-Richardson 20 coefficient value was 0.63 for the total score, with a Spearman-Brown value for the half-split of 0.63. Individuals with lower HbA1c values scored significantly higher on knowledge about hypoglycaemia prevention and correction (r = - 0.269, p = 0.007) and on the total questionnaire score (r = - 0.205, p = 0.041). A higher TIR and a lower TAR were also associated with a higher total score (r = 0.274, p = 0.007 and r = - 0.274, p = 0.007, respectively). CONCLUSIONS: In this study, the AdultCarbQuiz questionnaire, translated and culturally adapted to the Portuguese context, proved to be a useful tool in assessing knowledge about CH counting in patients with T1DM, allowing to optimize, individually, the therapeutic strategy in consultation.

Complex craniosynostosis in Pitt-Hopkins Syndrome: Case report in twins.

INTRODUCTION: Pitt-Hopkins syndrome (PTHS) is a rare genetic syndrome associated with neurodevelopmental disorders and craniofacial dysmorphisms caused by variations in the TCF4 transition factor. The aim of this article is to report the case of two twin infants diagnosed with PTHS, confirmed by the identification of a heterozygous pathogenic variant in the TCF4 gene through DNA extracted from a buccal swab. CASE PRESENTATION: Both infants presented with craniofacial asymmetry with a metopic crest and cranial deformity. During the diagnostic investigation, computed tomography with three-dimensional reconstruction of the skull showed premature fusion of the left coronal and metopic sutures in both twins. They underwent craniofacial reconstruction at the 9th month of age using a combination of techniques. The postoperative outcomes were satisfactory in both cases. CONCLUSION: To the best of our knowledge, this is the first case report to describe the occurrence of complex craniosynostosis (CCS) in children with PTHS. Further studies are needed to determine whether the co-occurrence of PTHS and CCS described here indicates an association or is explained by chance.

Frequency of CDH1, CTNNA1 and CTNND1 Germline Variants in Families with Diffuse and Mixed Gastric Cancer.

The most well-characterized hereditary form of gastric cancer is hereditary diffuse gastric cancer (HDGC), an autosomal dominant syndrome characterized by an increased risk of diffuse gastric and lobular breast cancer. HDGC is predominantly caused by germline pathogenic variants in the CDH1 gene, and more rarely in the CTNNA1 gene. Furthermore, the International Gastric Cancer Linkage Consortium (IGCLC) guidelines do not clarify whether or not mixed gastric cancer (with a diffuse component) should be considered in the HDGC genetic testing criteria. We aimed to evaluate the contribution of CTNNA1 and CTNND1 germline variants to HDGC. Additionally, we also intended to compare the frequencies of CDH1 and CTNNA1 (and eventually CTNND1) germline variants between patients with diffuse and mixed gastric carcinomas to evaluate if genetic testing for these genes should or should not be considered in patients with the latter. We analyzed the CDH1 gene in 67 cases affected with early-onset/familial mixed gastric carcinomas and the CTNNA1 and CTNND1 genes in 208 cases with diffuse or mixed gastric cancer who had tested negative for CDH1 pathogenic germline variants. A deleterious CTNNA1 germline variant was found in 0.7% (1/141) of diffuse gastric cancer patients meeting the 2020 IGCLC criteria, as compared to the rate of 2.8% of CDH1 deleterious variants found by us in this setting. No deleterious variants were found in CTNND1, but six variants of uncertain significance were identified in this gene. We did not find any pathogenic CDH1, CTNNA1 or CTNND1 variant in index patients with early-onset/familial mixed gastric cancer, so there is no evidence that supports including this tumor type in the testing criteria for germline variants in these genes. The role of the CTNND1 gene in inherited gastric cancer predisposition is still unclear.

Combined germline and tumor mutation signature testing identifies new families with NTHL1 tumor syndrome.

NTHL1 tumor syndrome is an autosomal recessive rare disease caused by biallelic inactivating variants in the NTHL1 gene and which presents a broad tumor spectrum. To contribute to the characterization of the phenotype of this syndrome, we studied 467 index patients by KASP assay or next-generation sequencing, including 228 patients with colorectal polyposis and 239 patients with familial/personal history of multiple tumors (excluding multiple breast/ovarian/polyposis). Three NTHL1 tumor syndrome families were identified in the group of patients with polyposis and none in patients with familial/personal history of multiple tumors. Altogether, we identified nine affected patients with polyposis (two of them diagnosed after initiating colorectal cancer surveillance) with biallelic pathogenic or likely pathogenic NTHL1 variants, as well as two index patients with one pathogenic or likely pathogenic NTHL1 variant in concomitance with a missense variant of uncertain significance. Here we identified a novel inframe deletion classified as likely pathogenic using the ACMG criteria, supported also by tumor mutational signature analysis. Our findings indicate that the NTHL1 tumor syndrome is a multi-tumor syndrome strongly associated with polyposis and not with multiple tumors without polyposis.

Implementation of upfront DPYD genotyping with a low-cost and high-throughput assay to guide fluoropyrimidine treatment in cancer patients.

OBJECTIVES: Genetic variants in the dihydropyrimidine dehydrogenase (DPYD ) gene are associated with reduced dihydropyrimidine dehydrogenase enzyme activity and can cause severe fluoropyrimidine-related toxicity. We assessed the frequency of the four most common and well-established DPYD variants associated with fluoropyrimidine toxicity and implemented a relatively low-cost and high-throughput genotyping assay for their detection. METHODS: This study includes 457 patients that were genotyped for the DPYD c.1129-5923C>G, c.1679T>G, c.1905 + 1G>A and c.2846A>T variants, either by Sanger sequencing or kompetitive allele specific PCR (KASP) technology. Of these, 172 patients presented toxicity during treatment with fluoropyrimidines (post-treatment group), and 285 were tested before treatment (pretreatment group). RESULTS: Heterozygous DPYD variants were identified in 7.4% of the entire series of 457 patients, being the c.2846A>T the most frequent variant. In the post-treatment group, 15.7% of the patients presented DPYD variants, whereas only 2.5% of the patients in the pretreatment group presented a variant. The KASP assays designed in this study presented 100% genotype concordance with the results obtained by Sanger sequencing. CONCLUSIONS: The combined assessment of the four DPYD variants in our population increases the identification of patients at high risk for developing fluoropyrimidine toxicity, supporting the upfront routine implementation of DPYD variant genotyping. Furthermore, the KASP genotyping assay described in this study presents a rapid turnaround time and relatively low cost, making upfront DPYD screening feasible in clinical practice.

Genetic landscape of homologous recombination repair genes in early-onset/familial prostate cancer patients.

Prostate cancer (PrCa) is one of the three most frequent and deadliest cancers worldwide. The discovery of PARP inhibitors for the treatment of tumors with deleterious variants in homologous recombination repair (HRR) genes has placed PrCa on the roadmap of precision medicine. However, the overall contribution of HRR genes to the 10%-20% of carcinomas arising in men with early-onset/familial PrCa has not been fully clarified. We used targeted next-generation sequencing (T-NGS) covering eight HRR genes (ATM, BRCA1, BRCA2, BRIP1, CHEK2, NBN, PALB2, and RAD51C) and an analysis pipeline querying both small and large genomic variations to clarify their global and relative contribution to hereditary PrCa predisposition in a series of 462 early-onset/familial PrCa cases. Deleterious variants were found in 3.9% of the patients, with CHEK2 and ATM being the most frequently mutated genes (38.9% and 22.2% of the carriers, respectively), followed by PALB2 and NBN (11.1% of the carriers, each), and finally by BRCA2, RAD51C, and BRIP1 (5.6% of the carriers, each). Using the same NGS data, exonic rearrangements were found in two patients, one pathogenic in BRCA2 and one of unknown significance in BRCA1. These results contribute to clarify the genetic heterogeneity that underlies PrCa predisposition in the early-onset and familial disease, respectively.

Diachronic profile of startup companies through social media.

Social media platforms have become powerful tools for startups, helping them find customers and raise funding. In this study, we applied a social media intelligence-based methodology to analyze startups' content and to understand how their communication strategies may differ during their scaling process. To understand if a startup's social media content reflects its current business maturation position, we first defined an adequate life cycle model for startups based on funding rounds and product maturity. Using Twitter as the source of information and selecting a sample of known Portuguese IT startups at different phases of their life cycle, we analyzed their Twitter data. After preprocessing the data, using latent Dirichlet allocation, topic modeling techniques enabled the categorization of the data according to the topics arising in the published contents of the startups, making it possible to discover that contents can be grouped into five specific topics: "Fintech and ML," "IT," "Business Operations," "Product/Service R&D," and "Bank and Funding." By comparing those profiles against the startup's life cycle, we were able to understand how contents change over time. This provided a diachronic profile for each company, showing that while certain topics remain prevalent in the startup's scaling, others depend on a particular phase of the startup's cycle. Our analysis revealed that startups' social media content differs along their life cycle, highlighting the importance of understanding how startups use social media at different stages of their development.

Potato Chips Byproducts as Feedstocks for Developing Active Starch-Based Films with Potential for Cheese Packaging.

The potato chip industry generates brownish frying residues, which are usually landfilled. While spent frying oil has value as biodiesel, the defatted brownish water-soluble extract (BrE) does not yet have an application. In this work, it was hypothesized that BrE can be a source of compounds for active packaging. BrE is composed of carbohydrates (66.9%), protein (5.7%), and a small amount of phenolics and esterified fatty acids. When incorporated into starch-based formulations and casted, BrE at 5%, 10%, and 15% w/w (dry starch weight) conferred a yellowish coloration while maintaining the transparency of neat films. The BrE increased the films' traction resistance, elasticity, and antioxidant activity while decreasing their hydrophilicity. Furthermore, starch/15% BrE-based films showed diminished water vapor and good UV-light barrier properties. Their contact with sliced cheese did not change the products' hardness during storage (14 days). Weight loss of the cheese was observed after 7 days of storage, stabilizing at 6.52%, contrary to the cheese packed in polyamide (PA)/polyethylene (PE), already used in food packaging. The cheese packed in the starch/15% BrE-based films showed a significant yellowish darkening and lower content of volatile oxidation products compared to the PA/PE. Therefore, BrE revealed to have compounds with the potential to tune the performance of starch-based films for food packaging.

Evaluation of PIK3CA mutations in advanced ER+/HER2-breast cancer in Portugal - U-PIK Project.

Background: Around 40% of ER+/HER2-breast carcinomas (BC) present mutations in the PIK3CA gene. Assessment of PIK3CA mutational status is required to identify patients eligible for treatment with PI3Kα inhibitors, with alpelisib currently the only approved tyrosine kinase inhibitor in this setting. U-PIK project aimed to conduct a ring trial to validate and implement the PIK3CA mutation testing in several Portuguese centers, decentralizing it and optimizing its quality at national level. Methods: Eight Tester centers selected two samples of patients with advanced ER+/HER2- BC and generated eight replicates of each (n = 16). PIK3CA mutational status was assessed in two rounds. Six centers used the cobas® PIK3CA mutation test, and two used PCR and Sanger sequencing. In parallel, two reference centers (IPATIMUP and the Portuguese Institute of Oncology [IPO]-Porto) performed PIK3CA mutation testing by NGS in the two rounds. The quality of molecular reports describing the results was also assessed. Testing results and molecular reports were received and analyzed by U-PIK coordinators: IPATIMUP, IPO-Porto, and IPO-Lisboa. Results: Overall, five centers achieved a concordance rate with NGS results (allele frequency [AF] ≥5%) of 100%, one of 94%, one of 93%, and one of 87.5%, considering the overall performance in the two testing rounds. NGS reassessment of discrepancies in the results of the methods used by the Tester centers and the reference centers identified one probable false positive and two mutations with low AF (1-3%, at the analytical sensitivity threshold), interpreted as subclonal variants with heterogeneous representation in the tissue sections processed by the respective centers. The analysis of molecular reports revealed the need to implement the use of appropriate sequence variant nomenclature with the identification of reference sequences (HGVS-nomenclature) and to state the tumor cell content in each sample. Conclusion: The concordance rates between the method used by each tester center and NGS validate the use of the PIK3CA mutational status test performed at these centers in clinical practice in patients with advanced ER+/HER2- BC.

Purification, Characterization, and Assessment of Antimicrobial Activity and Toxicity of Portulaca elatior Leaf Lectin (PeLL).

Lectins are carbohydrate-binding proteins with several bioactivities, including antimicrobial properties. Portulaca elatior is a species found at Brazilian Caatinga and data on the biochemical composition of this plant are scarce. The present work describes the purification of P. elatior leaf lectin (PeLL) as well as the assessment of its antimicrobial activity and toxicity. PeLL, isolated by chromatography on a chitin column, had native liquid charge and subunit composition evaluated by electrophoresis. Hemagglutinating activity (HA) of PeLL was determined in the presence of carbohydrates or divalent cations, as well as after heating and incubation at different pH values. Changes in the lectin conformation were monitored by evaluating intrinsic tryptophan fluorescence and using the extrinsic probe bis-ANS. Antimicrobial activity was evaluated against Pectobacterium strains and Candida species. The minimal inhibitory (MIC), bactericidal (MBC), and fungicidal (MFC) concentrations were determined. Finally, PeLL was evaluated for in vitro hemolytic activity in human erythrocytes and in vivo acute toxicity in mice (5 and 10 mg/kg b.w. per os). PeLL (pI 5.4; 20 kDa) had its HA was inhibited by mannose, galactose, Ca2+, Mg2+, and Mn2+. PeLL HA was resistant to heating at 100 °C, although conformational changes were detected. PeLL was more active in the acidic pH range, in which no conformational changes were observed. The lectin presented MIC and MBC of 0.185 and 0.74 µg/mL for all Pectobacterium strains, respectively; MIC of 1.48 µg/mL for C. albicans, C. tropicalis, and C. krusei; MIC and MFC of 0.74 and 2.96 µg/mL for C. parapsilosis. No hemolytic activity or signs of acute toxicity were observed in the mice. In conclusion, a new, low-toxic, and thermostable lectin was isolated from P. elatior leaves, being the first plant compound to show antibacterial activity against Pectobacterium.

Avaliação imunológica dos camundongos geneticamente selecionados para diferente capacidade de produção de anticorpo

INTRODUCTION: The potentiality of the inflammatory response or the specific immune response of the individual are complex characteristics that vary continuously and have a normal distribution in a heterogeneous population, since they are under polygenic control. Aiming at the study of the genetic regulation of specific immunity, our laboratory developed strains of heterogeneous mice genetically selected for high – HIII and low – LIII ability to produce antibodies to certain antigens through the process of genetic selection bidirectional. For some genetic and immunological studies, isogenic mice must be used. Therefore, we produced isogenic trunks of these parental lines in the vivarium of the Immunogenetics laboratory of the Butantan Institute, which after the selection process were subjected to inbreeding. OBJECTIVE: Evaluate the ability to produce antibodies as well as cells – B and T lymphocytes (CD4 and CD8) after stimulating the immune system with diphtheria anatoxin, comparing isogenic trunks with parental lineages. METHODOLOGY: Isogenic mice from Stem A (High) and Stem E (Low) and heterogeneous (HIII and LIII) were immunized with diphtheria anatoxin. The primary and secondary humoral immune response was evaluated in serum by quantifying the production of anti-diphtheria IgG antibodies using the ELISA technique. For the characterization of the T and B lymphocyte cell response profile, the spleen and lymph nodes were collected and the cells were identified by surface markers (CD4, CD8, B220) and evaluated by flow cytometry. RESULTS: The results show significant increases in the concentration of antibodies produced by the groups when comparing normal serum with primary and secondary responses. The production of antibodies in the secondary response was higher in animals from the HIII and Stem A strains compared to the other groups. In the cellular response, a significant increase in T lymphocytes (CD4+and CD8+) and B lymphocytes was seen in the lymph node in the experimental HIII group compared to the other groups, however in the spleen a significantly increased T cell response was detected in the immunized animals in the LIII group. Regarding the isogenic trunks, the animals from Stem A showed an increase in CD4+ T lymphocytes when compared to the experimental Stem E in splenic cells. CONCLUSIONS: Animals selected for high antibody production capacity showed greater responses both in IgG secretion in the secondary response and in the higher frequency of T and B cells in the lymph nodes. It is important to in-depth study of the modulation mechanism of the immune response in these strains to understand the multi-specific genetic effects against certain antigens.

INTRODUÇÃO: A potencialidade da resposta inflamatória ou da resposta imune específica do indivíduo são características complexas que variam de forma contínua e têm uma distribuição normal numa população heterogênea, pois estão sob controle poligênico. Visando o estudo da regulação genética da imunidade específica nosso laboratório desenvolveu linhagens de camundongos heterogênicos geneticamente selecionadas para alta (“High” - HIII) e baixa (“Low” - LIII) capacidade de produção de anticorpos a certos antígenos através do processo de seleção genética bidirecional. Para alguns estudos genéticos e imunológicos devem ser utilizados camundongos isogênicos, assim, produzimos no Biotério do laboratório de Imunogenética do Instituto Butantan troncos isogênicos destas linhagens parentais, que após o processo seletivo foram submetidas a cruzamentos consanguíneos. OBJETIVO: Avaliar a capacidade de produção de anticorpos assim como as células – linfócitos B e T (CD4+ e CD8+) após o estímulo do sistema imune com anatoxina diftérica comparando os troncos isogênicos com as linhagens parentais. METODOLOGIA: Camundongos isogênicos (High – Tronco A e Low – Tronco E) e heterogênicos (HIII e LIII) foram imunizados com anatoxina diftérica. A resposta imune humoral primária e secundária foram avaliadas no soro pela quantificação da produção de anticorpos IgG antidiftéricos através da técnica de ELISA. Para a caracterização do perfil de resposta celular de linfócitos T e B, foram coletados o baço e linfonodos e as células foram identificadas por marcadores de superfície (CD4, CD8, B220) e avaliadas por citometria de fluxo. RESULTADOS: Os resultados mostram aumentos significativos na concentração de anticorpos produzidos pelos grupos quando comparadas o soro normal com as respostas primária e secundária. A produção de anticorpos na resposta secundária foi maior nos animais das linhagens HIII e Tronco A em relação aos outros grupos. Na resposta celular foi visualizado no linfonodo um aumento significante de linfócitos T (CD4+e CD8+) e linfócitos B no grupo HIII experimental em relação aos outros grupos, entretanto no baço a resposta significativamente aumentada das células T foi detectada nos animais imunizados do grupo LIII. Em relação aos troncos isogênicos, os animais do Tronco A apresentaram um aumento de linfócitos T CD4+ quando comparados ao Tronco E experimental nas células esplênicas. CONCLUSÕES: Os animais selecionados para alta capacidade de produção de anticorpos apresentaram respostas maiores tanto na secreção de IgG na resposta secundária como na maior frequência de células T e B nos linfonodos. Sendo importante o estudo aprofundado do mecanismo de modulação da resposta imune nessas linhagens para entendimento de efeitos genéticos multiespecíficos frente a determinados antígenos.

Assessment of main complications of regional anesthesia recorded in an acute pain unit in a tertiary care university hospital: a retrospective cohort

Abstract Background Regional anesthesia has been increasingly used. Despite its low number of complications, they are associated with relevant morbidity. This study aims to evaluate the incidence of complications after neuraxial block and peripheral nerve block. Methods A retrospective cohort study was conducted, and data related to patients submitted to neuraxial block and peripheral nerve block at a tertiary university hospital from January 1, 2011 to December 31, 2017 were analyzed. Results From 10,838 patients referred to Acute Pain Unit, 1093(10.1%) had side effects or complications: 1039 (11.4%) submitted to neuraxial block and 54 (5.2%) to peripheral nerve block. The most common side effects after neuraxial block were sensory (48.5%) or motor deficits (11.8%), nausea or vomiting (17.5%) and pruritus (8.0%); The most common complications: 3 (0.03%) subcutaneous cell tissue hematoma, 3 (0.03%) epidural abscesses and 1 (0.01%) arachnoiditis. 204 of these patients presented sensory or motor deficits at hospital discharge and needed follow-up. Permanent peripheral nerve injury after neuraxial block had an incidence of 7.7:10,000 (0.08%). The most common side effects after peripheral nerve block were sensory deficits (52%) and 21 patients maintained follow-up due to symptoms persistence after hospital discharge. Conclusion Although we found similar incidences of side effects or even lower than those described, major complications after neuraxial block had a higher incidence, particularly epidural abscesses. Despite this, other serious complications, such as spinal hematoma and permanent peripheral nerve injury, are still rare.

Establishment and characterization of a novel ovarian high-grade serous carcinoma cell line-IPO43.

BACKGROUND: Epithelial ovarian cancer (EOC) is an aggressive and lethal malignancy and novel EOC cell lines with detailed characterization are needed, to provide researchers with diverse helpful resources to study EOC biological processes and cancer experimental therapies. METHODS: The IPO43 cell line was established from the ascitic fluid of a patient with a diagnosis of high-grade serous carcinoma (HGSC) of the ovary, previously treated with chemotherapy. Cell immortalization was achieved in 2D cell culture and growth obtained in 2D and 3D cell cultures. The characterization of immortalized cells was done by immunocytochemistry, flow cytometry, cell proliferation, chromosomal Comparative Genomic Hybridization (cCGH), STR profile and Next Generation Sequencing (NGS). RESULTS: Characterization studies confirmed that IPO43 cell line is of EOC origin and maintains morphological and molecular features of the primary tumor. cCGH analysis showed a complex profile with gains and losses of specific DNA regions in both primary ascitic fluid and cell line IPO43. The cell line was successfully grown in a 3D system which allows its future application in more complex assays than those performed in 2D models. IPO43 cell line is resistant to standard drug treatment in vitro. CONCLUSIONS: IPO43 is available for public research and we hope it can contribute to enrich the in vitro models addressing EOC heterogeneity, being useful to investigate EOC and to develop new therapeutic modalities.

KRAS and NRAS mutational analysis in plasma ctDNA from patients with metastatic colorectal cancer by real-time PCR and digital PCR.

PURPOSE: Mutations in the KRAS and NRAS (RAS) genes are negative predictors of response to anti-EGFR therapy in metastatic colorectal cancer (mCRC). The detection of mutations in circulating tumor DNA (ctDNA) has emerged as a less invasive strategy to assess the molecular profile of mCRC patients. We aimed to perform RAS mutational analysis in ctDNA from mCRC patients using BEAMing Digital PCR (OncoBEAM) and Idylla ctDNA qPCR and evaluate the concordance rate with RAS mutational status in tumor tissue and between these two methodologies with different limits of detection. METHODS: Blood samples were collected from 47 mCRC patients previously tested for RAS mutations in tumor tissue. DNA was extracted from plasma using the QIAamp Circulating Nucleic Acid Kit, and RAS mutation analysis was conducted using OncoBEAM RAS CRC and Idylla ctRAS assays. RESULTS: The overall agreement between tumor tissue and ctDNA analyses was 83% and 78.7% using the OncoBEAM and Idylla assays, respectively, with the concordance being 96.2% and 88.5% in naive treatment patients. The overall agreement between OncoBEAM and Idylla ctDNA analyses was 91.7%. CONCLUSIONS: Analysis of ctDNA is a viable strategy for clinical management of mCRC patients. Although the OncoBEAM assay sensitivity is somewhat higher, the fully automated Idylla platform also has good performance, while being cheaper and much less labor-intensive, for the detection of RAS mutations in plasma, either at diagnosis or after progression when considering anti-EGFR treatment rechallenge.

Expression Profiling in Ovarian Cancer Reveals Coordinated Regulation of BRCA1/2 and Homologous Recombination Genes.

Predictive biomarkers are crucial in clarifying the best strategy to use poly(ADP-ribose) polymerase inhibitors (PARPi) for the greatest benefit to ovarian cancer patients. PARPi are specifically lethal to cancer cells that cannot repair DNA damage by homologous recombination (HR), and HR deficiency is frequently associated with BRCA1/2 mutations. Genetic tests for BRCA1/2 mutations are currently used in the clinic, but results can be inconclusive due to the high prevalence of rare DNA sequence variants of unknown significance. Most tests also fail to detect epigenetic modifications and mutations located deep within introns that may alter the mRNA. The aim of this study was to investigate whether quantitation of BRCA1/2 mRNAs in ovarian cancer can provide information beyond the DNA tests. Using the nCounter assay from NanoString Technologies, we analyzed RNA isolated from 38 ovarian cancer specimens and 11 normal fallopian tube samples. We found that BRCA1/2 expression was highly variable among tumors. We further observed that tumors with lower levels of BRCA1/2 mRNA showed downregulated expression of 12 additional HR genes. Analysis of 299 ovarian cancer samples from The Cancer Genome Atlas (TCGA) confirmed the coordinated expression of BRCA1/2 and HR genes. To facilitate the routine analysis of BRCA1/2 mRNA in the clinical setting, we developed a targeted droplet digital PCR approach that can be used with FFPE samples. In conclusion, this study underscores the potential clinical benefit of measuring mRNA levels in tumors when BRCA1/2 DNA tests are negative or inconclusive.

Assessment of main complications of regional anesthesia recorded in an acute pain unit in a tertiary care university hospital: a retrospective cohort.

BACKGROUND: Regional anesthesia has been increasingly used. Despite its low number of complications, they are associated with relevant morbidity. This study aims to evaluate the incidence of complications after neuraxial block and peripheral nerve block. METHODS: A retrospective cohort study was conducted, and data related to patients submitted to neuraxial block and peripheral nerve block at a tertiary university hospital from January 1, 2011 to December 31, 2017 were analyzed. RESULTS: From 10,838 patients referred to Acute Pain Unit, 1093(10.1%) had side effects or complications: 1039 (11.4%) submitted to neuraxial block and 54 (5.2%) to peripheral nerve block. The most common side effects after neuraxial block were sensory (48.5%) or motor deficits (11.8%), nausea or vomiting (17.5%) and pruritus (8.0%); The most common complications: 3 (0.03%) subcutaneous cell tissue hematoma, 3 (0.03%) epidural abscesses and 1 (0.01%) arachnoiditis. 204 of these patients presented sensory or motor deficits at hospital discharge and needed follow-up. Permanent peripheral nerve injury after neuraxial block had an incidence of 7.7:10,000 (0.08%). The most common side effects after peripheral nerve block were sensory deficits (52%) and 21 patients maintained follow-up due to symptoms persistence after hospital discharge. CONCLUSION: Although we found similar incidences of side effects or even lower than those described, major complications after neuraxial block had a higher incidence, particularly epidural abscesses. Despite this, other serious complications, such as spinal hematoma and permanent peripheral nerve injury, are still rare.

Development of the prickly pear cactus Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro in response to the replacement of potassium nitrate for a commercial kno 3 fertilizer / Desenvolvimento da palma Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro em resposta à substituição de nitrato de potássio (P.A.) por fertilizante comercial KNO 3

ABSTRACT: In micropropagation, potassium nitrate (KNO3), an ACS reagent grade chemical, used in the preparation of growing mediums is expensive and its procurement depends on bureaucratic procedures, as it is controlled by the Brazilian Army. This research to assessed the effect of replacing the ACS KNO3 for a commercially available fertilizer (KNO3- based) on the micropropagation of the prickly pear cactus (Opuntia stricta (Haw.) Haw. cv. Elephant Ear. Treatments used six different fertilizer concentrations (0, 0.5, 1, 1.5, 2 and 2.5 g L-1) and a control consisting of 1.9 g L-1 KNO3, as shown in the MS salts. The survival, size and number of sprouts and the value of fresh biomass were evaluated. After seedling acclimation, we assessed the survival, number of sprouts, length, and number of roots, racket formation, average fresh biomass mass, macronutrient absorption and morphological changes of the seedlings. Explants inoculated with fertilizers at concentrations of 0.0; 2.0 and 2.5 g L-¹ did not grow. The response of explants at concentrations of 0.5 and 1.5 g L-1 of the fertilizer were the same as those developed in a KNO3 medium, and at a concentration of 1.0 g L-1, in all variables, the means were higher than those of the control medium. Therefore, it showed the feasibility of using fertilizers in the in vitro cultivation of the prickly pear cactus, which may remove bureaucratic barriers and reduce product costs by 99.12%.

RESUMO: Na micropropagação, o nitrato de potássio (KNO3), reagente puro para análise (P.A.), utilizado no preparo dos meios de cultura, possui custo elevado e a sua aquisição depende de trâmites burocráticos, por se tratar de substância controlada pelo Exército Brasileiro. O objetivo deste trabalho foi avaliar o efeito da substituição do KNO3 P.A. por fertilizante comercial (com fonte de KNO3), encontrado livremente no comércio, na micropropagação de palma (Opuntia stricta (Haw.) Haw. cv Orelha de Elefante. Os tratamentos foram de seis concentrações do fertilizante (0; 0,5; 1; 1,5; 2 e 2,5 g L-1) e um controle constituído de 1,9 g L-1 de reagente KNO3, conforme mostrado nos sais MS. Avaliou-se a sobrevivência, tamanho e número de brotações do explante, e o valor da biomassa fresca. Após a aclimatização das mudas avaliou-se a sobrevivência, número de brotações, comprimento da parte aérea, número de raízes, formação da raquete, massa média da biomassa fresca, absorção de macronutrientes e alterações morfológicas das mudas. Os explantes inoculados em meio com fertilizantes nas concentrações de 0,0; 2,0 e 2,5 g L-¹ não se desenvolveram. A resposta dos explantes nas concentrações de 0,5 e 1,5 g L-1 do fertilizante foram iguais aos desenvolvidos em meio contendo KNO3, e na concentração de 1,0 g L-1, em todas as variáveis, as médias foram superiores em relação as do controle. Dessa forma, constatou-se a viabilidade do uso do fertilizante no cultivo in vitro da palma, o que propiciou a eliminação dos entraves burocráticos e redução no custo de 99,12% na compra do produto.

Percepção e práticas de pais/responsáveis sobre questões atuais da odontopediatria: um estudo piloto / Perceptions and practices of parents and guardians on current issues of pediatric dentistry: a pilot study

Objetivo: Investigar as percepções e atitudes de pais/responsáveis sobre cuidados relacionados à saúde bucal das crianças por meio de um estudo descritivo. Métodos: Foi aplicado um questionário semiestruturado aos pais/responsáveis de crianças atendidas nas clinicas de Odontopediatria da Pontifícia Universidade Católica de Minas Gerais (PUC-MG) em 78 pais/responsáveis. O questionário apresentava questões sociodemográficas, assim como questões específicas relacionadas aos cuidados e atitudes para a saúde bucal de seus filhos/crianças. Análise descritiva e exploratória, por meio de médias, frequências e porcentagens foram realizadas e ilustradas por meio de figuras. Resultados: Embora a maioria dos entrevistados tenha alegado já terem recebido informações a respeito dos cuidados bucais infantis, pais/responsáveis consideraram o momento propício para a primeira visita ao dentista somente após o 1º ano de vida, período inadequado para o aleitamento materno, além da utilização inadequada de chupeta e mamadeira por longos períodos e do período para a inserção do hábito da escovação. Além disso, temáticas relacionadas à erosão dentária, teste da linguinha e a importância do íon fluoreto para a saúde bucal foram os assuntos classificados com mais dúvidas. Conclusão: Em geral, as atitudes dos pais e responsáveis não se mostraram adequadas, apesar de possuírem informações prévias no que tange a saúde bucal das crianças. Com isso, verifica-se a necessidade de enfatizar mais a fundo questões referentes a esta temática e diferentes meios para a difusão deste tipo de informação.

Aim: To investigate the perceptions and attitudes of parents/guardians about oral health care for children through a descriptive/cross-sectional study. Methods: A semi-structured questionnaire was applied to 78 parents/guardians of children seen at the Pediatric Dentistry Clinics of the Pontific Catholic University of Minas Gerais (PUC-MG). The questionnaire presented sociodemographic questions, as well as specific questions related to the care and attitudes towards the oral health of their children. Exploratory and descriptive analyses by means of means, frequencies,and percentages were performed and illustrated in figures. Results: Although most of the interviewees claimed to have already received information regarding children's oral care, they considered the proper time for the first visit to the dentist only after the 1st year of life, an inadequate period for breastfeeding, as well as the inappropriate use of pacifiers and bottles for long periods and an improper period for the insertion of the habit of tooth brushing. Furthermore, issues related to dental erosion, the tongue test, and the importance of fluoride ions for oral health were the subjects classified with the most doubts. Conclusion: In general, the attitudes of parents and guardians proved to be inappropriate, although they have previous information regarding the oral health of children. Thus, there is a need to offer a more in-depth emphasis on issues related to this theme and different means through which to disseminate this type of information.