22q11 Copy Number Variations in a Brazilian Cohort of Children with Congenital Heart Disorders.

Introduction: Congenital heart disease (CHD) is the most common type of congenital defect reported to be one of the leading causes of mortality in the first year of life. Microdeletion and microduplication syndromes (MMS) are associated with cardiac malformations. Understanding which genetic factors are involved in these conditions directly impacts treatment decisions. We aimed to identify the occurrence of genetic alterations and their association with MMS in CHD pediatric patients evaluated in a reference service of Southern Brazil. Methods: Participants were recruited during 2010 in the intensive care unit of a pediatric hospital. MMs and regions of chromosome 22 were screened by SALSA MLPA Probemix P245 Microdeletion Syndromes-1A kit for detection of copy number variations (CNVs). Results: MMS were detected in 11 from 207 patients (5.3%). Heterozygous deletion in the 22q11.2 chromosome region was the most prevalent CNV (5 from 11 patients). Also, atypical RTDR1 deletion and 22q11.2 duplication were detected. MLPA was able to reveal microdeletions in SNRPN and NF1 genes in patients with a normal karyotype and FISH. Conclusion: Our study reports the prevalence and variability of genomic alterations associated with MMS in CHD pediatric patients. The results by MLPA are of great help in planning and specialized care.

Which outcomes are most important to measure in patients with COVID-19 and how and when should these be measured? Development of an international standard set of outcomes measures for clinical use in patients with COVID-19: a report of the International Consortium for Health Outcomes Measurement (ICHOM) COVID-19 Working Group.

OBJECTIVES: The COVID-19 pandemic has resulted in widespread morbidity and mortality with the consequences expected to be felt for many years. Significant variation exists in the care even of similar patients with COVID-19, including treatment practices within and between institutions. Outcome measures vary among clinical trials on the same therapies. Understanding which therapies are of most value is not possible unless consensus can be reached on which outcomes are most important to measure. Furthermore, consensus on the most important outcomes may enable patients to monitor and track their care, and may help providers to improve the care they offer through quality improvement. To develop a standardised minimum set of outcomes for clinical care, the International Consortium for Health Outcomes Measurement (ICHOM) assembled a working group (WG) of 28 volunteers, including health professionals, patients and patient representatives. DESIGN: A list of outcomes important to patients and professionals was generated from a systematic review of the published literature using the MEDLINE database, from review of outcomes being measured in ongoing clinical trials, from a survey distributed to patients and patient networks, and from previously published ICHOM standard sets in other disease areas. Using an online-modified Delphi process, the WG selected outcomes of greatest importance. RESULTS: The outcomes considered by the WG to be most important were selected and categorised into five domains: (1) functional status and quality of life, (2) mental functioning, (3) social functioning, (4) clinical outcomes and (5) symptoms. The WG identified demographic and clinical variables for use as case-mix risk adjusters. These included baseline demographics, clinical factors and treatment-related factors. CONCLUSION: Implementation of these consensus recommendations could help institutions to monitor, compare and improve the quality and delivery of care to patients with COVID-19. Their consistent definition and collection could also broaden the implementation of more patient-centric clinical outcomes research.

ABC2-SPH risk score for in-hospital mortality in COVID-19 patients: development, external validation and comparison with other available scores.

OBJECTIVES: The majority of available scores to assess mortality risk of coronavirus disease 2019 (COVID-19) patients in the emergency department have high risk of bias. Therefore, this cohort aimed to develop and validate a score at hospital admission for predicting in-hospital mortality in COVID-19 patients and to compare this score with other existing ones. METHODS: Consecutive patients (≥ 18 years) with confirmed COVID-19 admitted to the participating hospitals were included. Logistic regression analysis was performed to develop a prediction model for in-hospital mortality, based on the 3978 patients admitted between March-July, 2020. The model was validated in the 1054 patients admitted during August-September, as well as in an external cohort of 474 Spanish patients. RESULTS: Median (25-75th percentile) age of the model-derivation cohort was 60 (48-72) years, and in-hospital mortality was 20.3%. The validation cohorts had similar age distribution and in-hospital mortality. Seven significant variables were included in the risk score: age, blood urea nitrogen, number of comorbidities, C-reactive protein, SpO2/FiO2 ratio, platelet count, and heart rate. The model had high discriminatory value (AUROC 0.844, 95% CI 0.829-0.859), which was confirmed in the Brazilian (0.859 [95% CI 0.833-0.885]) and Spanish (0.894 [95% CI 0.870-0.919]) validation cohorts, and displayed better discrimination ability than other existing scores. It is implemented in a freely available online risk calculator (https://abc2sph.com/). CONCLUSIONS: An easy-to-use rapid scoring system based on characteristics of COVID-19 patients commonly available at hospital presentation was designed and validated for early stratification of in-hospital mortality risk of patients with COVID-19.

GATA 4 Deletions Associated with Congenital Heart Diseases in South Brazil.

The normal development of the heart comprises a highly regulated machinery of genetic events, involving transcriptional factors. Congenital heart disease (CHD), have been associated with chromosomal abnormalities and copy number variants (CNVs). Our goal was to investigate through the multiplex ligation-dependent probe amplification (MLPA) technique, the presence of CNVs in reference genes for normal cardiac development in patients with CHD. GATA4 , NKX2-5 , TBX5 , BMP4 , and CRELD1 genes and 22q11.2 chromosome region were analyzed in 207 children with CHD admitted for the first time in a cardiac intensive care unit from a pediatric hospital. CNVs were detected in seven patients (3.4%): four had a 22q11.2 deletion (22q11DS) (1.9%), two had a GATA4 deletion (1%) and one had a 22q11.2 duplication (0.5%). No patients with CNVs in the NKX2-5 , TBX5 , BMP4 , and CRELD1 genes were identified. GATA4 deletions appear to be present in a significant number of CHD patients, especially those with septal defects, persistent left superior vena cava, pulmonary artery abnormalities, and extracardiac findings. GATA4 screening seems to be more effective when directed to these CHDs. The investigation of CNVs in GATA4 and 22q11 chromosome region in patients with CHD is important to anticipating the diagnosis, and to contributing to family planning.

Fetal anencephaly with umbilical cord attached to cerebrovasculosa area: autopsy report / Anencefalia fetal com cordão umbilical aderido à área cerebrovasculosa: relato da autópsia

ABSTRACT Our aim was to report the unusual case of an anencephalic fetus that was born with the umbilical cord attached to its cephalic pole. The patient was a 16-year-old pregnant woman. Ultrasound evaluation at 17 weeks revealed a single fetus without cranial vault and brain parenchyma, compatible with anencephaly. Postnatal evaluation, through an autopsy, was consistent with this diagnosis. Moreover, it was observed that the umbilical cord was attached to the area cerebrovasculosa and there was the presence of a fibrous tissue, suggestive of an amniotic band. We have found only one similar case described in the literature.

RESUMEN Reportamos el caso poco usual de un feto anencéfalo nacido con el cordón umbilical adherido al area cerebrovasculosa. La embarazada tenía 16 años de edad. La ecografía de la semana 17 de gestación mostró un feto sin bóveda craneal y parénquima cerebral, compatible con anencefalia. La evaluación posnatal, mediante autopsia, estaba de acuerdo a ese diagnóstico. Además, se ha notado que el cordón umbilical estaba adherido al area cerebrovasculosa, puesto que había presencia de tejido fibroso, sugestivo de banda amniótica. Encontramos solo un caso descrito en la literatura.

RESUMO Relatamos um caso incomum de feto com anencefalia nascido com o cordão umbilical aderido à área cerebrovasculosa. A gestante tinha 16 anos de idade. A avaliação pela ultrassonografia, na 17ª semana de gestação, revelou feto sem calota craniana e parênquima cerebral, compatível com anencefalia. A avaliação pós-natal, através da autópsia, foi concordante com esse diagnóstico. Ademais, notou-se que o cordão umbilical estava aderido à área cerebrovasculosa, visto que havia a presença de tecido fibroso, sugestivo de uma banda amniótica. Encontramos apenas um caso semelhante descrito na literatura.

Description of clinical aspects and microscopy of the hair shaft of a carrier of familial monilethrix / Descrição dos aspectos clínicos e da microscopia da haste capilar de um portador de monilétrix familiar

ABSTRACT Monilethrix is a genetic condition that affects the hair shaft. We describe a family with this disease, focusing on its clinical aspects and microscopic hair characteristics. The patient was a 10-year-old female with history of hypotrichosis. In addition to diffuse alopecia, there was brittle hair, with ruptures in the hair shaft at different levels. The hair had a nodular appearance at naked eye. Other family members had the same symptoms, what indicates an autosomal dominant pattern of inheritance. Microscopic analysis revealed capillary fibers with areas of elliptical nodular appearance interspersed with regions of dystrophic constriction.

RESUMO A monilétrix é uma condição genética que acomete a haste capilar. Descrevemos uma família com essa doença, enfocando seus aspectos clínicos e as características microscópicas do cabelo. A paciente era do sexo feminino, 10 anos de idade, e apresentava história de hipotricose. Além da alopecia difusa, notava-se um cabelo quebradiço, com rupturas na haste capilar em diferentes níveis. Os cabelos possuíam um aspecto nodular a olho nu. Outros membros da família apresentavam os mesmos sintomas, o que indica um padrão de herança autossômica dominante. A análise microscópica revelou fibras capilares com áreas de aparência nodular elíptica, intercaladas por regiões de constrição distrófica.

The role of cytogenetics and molecular biology in the diagnosis, treatment and monitoring of patients with chronic myeloid leukemia / O papel da citogenética e da biologia molecular no diagnóstico, no tratamento e no monitoramento de pacientes com leucemia mieloide crônica

ABSTRACT Chronic myeloid leukemia (CML) is the most common myeloproliferative disorder among chronic neoplasms. The history of this disease joins with the development of cytogenetic analysis techniques in human. CML was the first cancer to be associated with a recurrent chromosomal alteration, a reciprocal translocation between the long arms of chromosomes 9 and 22 - Philadelphia chromosome. This work is an updated review on CML, which highlights the importance of cytogenetics analysis in the continuous monitoring and therapeutic orientation of this disease. The search for scientific articles was carried out in the PubMed electronic database, using the descriptors "leukemia", "chronic myeloid leukemia", "treatment", "diagnosis", "karyotype" and "cytogenetics". Specialized books and websites were also included. Detailed cytogenetic and molecular monitoring can assist in choosing the most effective drug for each patient, optimizing the treatment. Cytogenetics plays a key role in the detection of chromosomal abnormalities associated with malignancies, as well as the characterization of new alterations that allow more research and increase knowledge about the genetic aspects of these diseases. The development of new drugs, through the understanding of the molecular mechanisms involved, will allow a possible improvement in the survival of these patients.

RESUMO Leucemia mieloide crônica (LMC) é a desordem mieloproliferativa mais comum entre as neoplasias crônicas. A história dessa doença se alia ao desenvolvimento de técnicas de análise citogenética em humanos. Foi o primeiro câncer a ser associado a uma alteração cromossômica recorrente, uma translocação recíproca entre os braços longos do cromossomo 9 e 22 - o cromossomo Philadelphia. Este trabalho é uma revisão atualizada sobre LMC, o qual destaca a importância da análise citogenética no monitoramento contínuo e na orientação terapêutica dessa doença. A pesquisa de artigos científicos foi realizada no banco de dados PubMed, usando os descritores "leucemia", "leucemia mieloide crônica", "tratamento", "diagnóstico", "cariótipo" e "citogenética". Livros e sites especializados também foram incluídos. O monitoramento citogenético e molecular detalhado pode auxiliar na escolha do medicamento mais efetivo para cada paciente, otimizando seu tratamento. A citogenética desempenha um papel fundamental na detecção de anormalidades cromossômicas associadas a malignidades, bem como na caracterização de novas alterações que permitem mais pesquisas e ampliação do conhecimento sobre os aspectos genéticos dessas doenças. O desenvolvimento de novas drogas, através da compreensão dos mecanismos moleculares envolvidos, permitirá uma possível melhora na sobrevida desses pacientes.

Increased levels of chitotriosidase in a patient with Alagille syndrome: association or coincidence? / Aumento dos níveis de quitotriosidase em um paciente com síndrome de Alagille: associação ou coincidência?

ABSTRACT We describe a case of a patient with Alagille syndrome (AS) presenting an increased level of the enzyme chitotriosidase (ChT), evaluating factors that could justify the relationship between AS and ChT. He was a male patient with cholestatic jaundice, facial dysmorphia and congenital heart disease who presented a brief septicemia. He underwent liver biopsy and analyses for inborn errors of metabolism that respectively showed ductopenia and increased levels of ChT. This increase could be potentially explained by inflammatory and infectious processes, or even by AS itself.

RESUMO Descrevemos o caso de um paciente do sexo masculino com síndrome de Alagille (SA), o qual manifestou aumento do nível da enzima quitotriosidase (ChT). Avaliamos os fatores que pudessem justificar a relação entre AS e ChT. O paciente apresentou icterícia colestática, tinha dismorfias faciais, cardiopatia congênita e manifestou um breve quadro de septicemia. Foi submetido à biópsia de fígado e análises para erros inatos do metabolismo que mostraram, respectivamente, ductopenia e aumento dos níveis de ChT. Esse aumento poderia ser potencialmente explicado por processos infecciosos e inflamatórios, ou mesmo pela própria SA.

Evaluation of the fetal urine sample through vesicocentesis: an approach to diagnostic and therapeutic application / Avaliação da urina fetal por meio da vesicocentese: uma abordagem com aplicação diagnóstica e terapêutica

ABSTRACT We report the case of a fetus with mega-bladder and suspected lower urinary tract obstruction (LUTO). The 20-week pregnancy ultrasound scan showed absence of amniotic fluid (anhydramnios), enlarged bladder, and narrowing of the urethra in the proximal region. At 21 weeks of gestational age, vesicocentesis was performed for relief of obstruction and analysis of biochemical of the fetal urine and karyotyping was carried out, which presented normal result (46,XY). This technique is indicated in cases of severe oligohydramnios or difficulty of placental access and has diagnostic and therapeutic function.

RESUMO Relatamos o caso de um feto com megabexiga e suspeita de obstrução do trato urinário inferior (LUTO). O exame ultrassonográfico realizado com 20 semanas de gestação mostrou líquido amniótico ausente (adramnia), bexiga com aumento de volume e estreitamento na região da uretra proximal. Na 21ª semana de gestação, foi realizada a vesicocentese para alívio e análise bioquímica e cariotípica da urina fetal, que apresentou resultado normal (46,XY). Essa técnica é indicada em casos de grave oligodramnia ou dificuldade de acesso placentário e tem função diagnóstica e terapêutica.

Report of a patient with fragile X syndrome unexpectedly identified by karyotype analysis / Relato de um paciente com a síndrome do X frágil identificada de forma inesperada por meio do cariótipo

ABSTRACT Fragile X syndrome is considered the main known cause of inherited learning disabilities and it is characterized by mutations in the FMR1 gene. Our aim was to report an unexpected detection of a patient with fragile X syndrome by GTG-Banding karyotype analysis (G-bands after trypsin and Giemsa). The karyotype analysis identified Xq27.3 fragility in 17% of the metaphases analyzed and in 54% when using TC 199, consistent with the cytogenetic diagnosis of the syndrome. This case was the sole one to present the fra(X) tests in the high-resolution karyotype analysis in our care service, contributing to future diagnoses of patients with history of developmental delay.

RESUMO A síndrome do X frágil é a principal causa conhecida de deficiência de aprendizagem herdada, caracterizada por mutações no gene FMR1. Relatamos a detecção inesperada de um paciente com síndrome do X frágil por meio de cariótipo de sangue periférico com bandamento GTG (bandamento G após tripsina e Giemsa). A análise cariotípica identificou fragilidade Xq27.3 em 17% das metáfases analisadas e em 54% quando utilizado TC 199, consistente com o diagnóstico citogenético da síndrome. Este caso foi o único a apresentar as provas de fra(X) no cariótipo de alta resolução em nosso serviço de atendimento, contribuindo para futuros diagnósticos de pacientes com história de atraso no desenvolvimento.

Applications of electron microscopy in health: the example of epidermolysis bullosa / Aplicações da microscopia eletrônica em saúde: o exemplo da epidermólise bolhosa

ABSTRACT We report the case of a patient with dystrophic epidermolysis bullosa (DEB) diagnosed by transmission electron microscopy (TEM), emphasizing the applications and importance of this technique in the health area. The patient was a male, the only child of young and non-consanguineous parents without similar cases in the family. The patient underwent a cutaneous biopsy in which TEM revealed sub-basal membrane involvement, confirming the diagnosis of DEB. Despite technological advances, TEM continues to play an important role in diagnosis and clinical research and is considered the best option for confirmation of diagnosis and subtypes of diseases such as epidermolysis bullosa (EB).

RESUMO Relatamos o caso de um paciente com epidermólise bolhosa distrófica (EBD) diagnosticado por microscopia eletrônica de transmissão (MET), destacando aplicações e importância desta técnica na área da saúde. Paciente do sexo masculino, filho único de pais jovens não consanguíneos, sem histórico de caso familial. O paciente foi submetido à biópsia cutânea, na qual a MET revelou comprometimento da membrana sub-basal, confirmando o diagnóstico de EBD. Apesar dos avanços tecnológicos, a MET continua tendo papel importante no diagnóstico e na pesquisa clínica, sendo considerada a melhor opção para a confirmação do diagnóstico e dos subtipos de doenças como a epidermólise bolhosa (EB).

Use of the fluid obtained by puncture of cystic hygroma: an alternative method for fetal karyotyping / Uso do fluido obtido por punção do higroma cístico: método alternativo para cariotipagem fetal

ABSTRACT The aim of our study aim was to report the case of a fetus with Turner syndrome (TS) diagnosed by karyotype from cystic hygroma (CH) fluid, highlighting the applications and importance of this procedure. First-trimester screening revealed an increased nuchal translucency measurement, cervical cystic hygroma and head and trunk subcutaneous edema. The presence of oligohydramnios prevented the performance of amniocentesis. We performed puncture of the CH for fetal karyotyping, which revealed X-chromosome monosomy (45,X), compatible with TS. Therefore, the use of CH fluid as an alternative sample for fetal karyotyping may be considered when conventional invasive procedures can not be performed.

RESUMO Relatamos o caso de um feto com síndrome de Turner (TS), diagnosticado por cariótipo de fluido do higroma cístico (CH), salientando as aplicações e a importância desse procedimento. Rastreio de primeiro trimestre revelou aumento da medida da translucência nucal, higroma cístico cervical e edema subcutâneo de cabeça e tronco. A presença de oligodrâmnio impediu a execução de amniocentese. Realizamos punção do CH para cariotipagem fetal, que revelou monossomia do cromossomo X (45,X), compatível com TS. Portanto, o uso de fluido do CH, como amostra alternativa para cariotipagem fetal, pode ser considerado quando procedimentos invasivos convencionais não podem ser realizados.