Steroid hormone profiles and molecular diagnostic tools in pediatric patients with non-CAH primary adrenal insufficiency

2022 ◽  
Author(s):  
Tuba Seven Menevse ◽  
Yasemin Kendir Demirkol ◽  
Busra Gurpinar Tosun ◽  
Elvan Bayramoglu ◽  
Melek Yildiz ◽  
...  
Keyword(s):  
Adrenal Insufficiency ◽  
Molecular Diagnosis ◽  
High Throughput Sequencing ◽  
Steroid Hormone ◽  
Genetic Diagnosis ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Unknown Etiology ◽  
Control Group ◽  
Primary Adrenal Insufficiency

Abstract Background There is a significant challenge of attributing specific diagnoses to patients with primary adrenal insufficiency of unknown etiology other than congenital adrenal hyperplasia (non-CAH PAI). Specific diagnoses per se may guide personalized treatment or may illuminate pathophysiology. Objective Investigation of the efficacy of steroid hormone profiles and high-throughput sequencing methods in establishing the etiology in non-CAH PAI of unknown origin. Design Paediatric patients with non-CAH PAI whose etiology could not be established by clinical and biochemical characteristics were enrolled. Genetic analysis was performed using targetedgene panel sequencing (TPS) and whole-exome sequencing (WES). Plasma adrenal steroids were quantified by liquid chromatography-mass spectrometry and compared to that of controls. Setting Eighteen pediatric endocrinology clinics. Patients Forty-one patients (17 females, median age: 3 months, range: 0-8 years) with non-CAH PAI of unknown etiology. Results A genetic diagnosis was obtained in 29 (70.7%) patients by TPS. Further molecular diagnosis could not be achieved by WES. Compared to healthy control group, patients showed lower steroid concentrations, most significantly in cortisone, cortisol, and corticosterone (p<0.0001, area under the ROC curve: 0.96, 0.88, 0.87, respectively). Plasma cortisol<4 ng/mL, cortisone<11 ng/mL, and corticosterone<0.11 ng/mL had >95% specificity to ensure the diagnosis of non-CAH PAI of unknown etiology. Conclusion Steroid hormone profiles are highly sensitive for the diagnosis of non-CAH PAI of unknown etiology, while they are unlikely to point out a specific molecular diagnosis. TPS is an optimal approach in the molecular diagnosis of these patients with high efficacy, while little additional benefit is expected from WES.

Molecular Diagnosis of Inherited Coagulation and Bleeding Disorders

2019 ◽  
Vol 45 (07) ◽  
pp. 695-707 ◽  
Author(s):  
José María Bastida ◽  
Rocío Benito ◽  
María Luisa Lozano ◽  
Ana Marín-Quilez ◽  
Kamila Janusz ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
High Throughput Sequencing ◽  
Clinical Care ◽  
Genetic Diagnosis ◽  
Multidisciplinary Teams ◽  
Molecular Defect ◽  
Bleeding Disorders ◽  
Whole Exome ◽  
Increased Risk ◽  
Risk Of Malignancy

AbstractDiagnosis of inherited bleeding disorders (IBDs) remains challenging, especially in the case of inherited platelet disorders, due to the heterogeneity of the clinical and laboratory phenotype, the limited specificity of platelet function tests, and the large number of potential culprit genes. Unraveling the underlying molecular defect provides the definitive diagnosis of IBDs, facilitating prognosis and clinical care, which are especially important for severe clinical syndromes and those that may be associated with an increased risk of malignancy. Until recently, Sanger sequencing of candidate genes has been the only method of molecular diagnosis, but this approach is time-consuming and costly and requires phenotype-based identification of any obvious candidate gene(s). Nowadays, high-throughput sequencing (HTS) allows the simultaneous and rapid investigation of multiple genes at a manageable cost. This HTS technology that includes targeted sequencing of prespecified genes, whole-exome sequencing, or whole-genome sequencing, is revolutionizing the genetic diagnosis of human diseases. Through its extensive implementation in research and clinical practice, HTS is rapidly improving the molecular characterization of IBDs. However, despite the availability of this powerful approach, many patients still do not receive a diagnosis. As IBDs are complex and rare diseases, development of more advanced laboratory assays, improvements in bioinformatic pipelines, and the formation of multidisciplinary teams are encouraged to advance our understanding of IBDs.

scholarly journals Clinical Significance of Molecular Diagnostic Tools for Bacterial Bloodstream Infections: A Systematic Review

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Jean Pierre Rutanga ◽  
Therese Nyirahabimana
Keyword(s):  
Systematic Review ◽  
Antibiotic Therapy ◽  
Clinical Significance ◽  
Molecular Diagnosis ◽  
Accurate Diagnosis ◽  
Molecular Techniques ◽  
Diagnostic Methods ◽  
Diagnosis And Treatment ◽  
Molecular Diagnostic ◽  
Diagnostic Tools

Bacterial bloodstream infection (bBSI) represents any form of invasiveness of the blood circulatory system caused by bacteria and can lead to death among critically ill patients. Thus, there is a need for rapid and accurate diagnosis and treatment of patients with septicemia. So far, different molecular diagnostic tools have been developed. The majority of these tools focus on amplification based techniques such as polymerase chain reaction (PCR) which allows the detection of nucleic acids (both DNA and small RNAs) that are specific to bacterial species and sequencing or nucleic acid hybridization that allows the detection of bacteria in order to reduce delay of appropriate antibiotic therapy. However, there is still a need to improve sensitivity of most molecular techniques to enhance their accuracy and allow exact and on time antibiotic therapy treatment. In this regard, we conducted a systematic review of the existing studies conducted in molecular diagnosis of bBSIs, with the main aim of reporting on clinical significance and benefits of molecular diagnosis to patients. We searched both Google Scholar and PubMed. In total, eighteen reviewed papers indicate that shift from conventional diagnostic methods to molecular tools is needed and would lead to accurate diagnosis and treatment of bBSI.

The SEeMORE strategy: single-tube electrophoresis analysis-based genotyping to detect monogenic diseases rapidly and effectively from conception until birth

2017 ◽  
Vol 56 (1) ◽  
Author(s):  
Federica Cariati ◽  
Maria Savarese ◽  
Valeria D’Argenio ◽  
Francesco Salvatore ◽  
Rossella Tomaiuolo
Keyword(s):  
Prenatal Diagnosis ◽  
Molecular Diagnosis ◽  
Biological Sample ◽  
Genetic Diagnosis ◽  
Disease Type ◽  
Molecular Diagnostic ◽  
Monogenic Disease ◽  
Laboratory Practice ◽  
Single Tube ◽  
Monogenic Diseases

AbstractBackground:The development of technologies that detect monogenic diseases in embryonic and fetal samples are opening novel diagnostic possibilities for preimplantation genetic diagnosis (PGD) and prenatal diagnosis (PND) thereby changing laboratory practice. Molecular diagnostic laboratories use different workflows for PND depending on the disease, type of biological sample, the presence of one or more known mutations, and the availability of the proband. Paternity verification and contamination analysis are also performed. The aim of this study was to test the efficacy of a single workflow designed to optimize the molecular diagnosis of monogenic disease in families at-risk of transmitting a genetic alteration.Methods:We used this strategy, which we designated “SEeMORE strategy” (Results:The results obtained with the SEeMORE strategy concurred with those obtained with traditional PND. In addition, this strategy has several advantages: (i) use of one or a few cells; (ii) reduction of the procedure to 1 day; and (iii) a reduction of at least 2–3-fold of the analytic cost.Conclusions:The SEeMORE strategy is effective for the molecular diagnosis of monogenic diseases, irrespective of the amount of starting material and of the disease mutation, and can be used for PND and PGD.

scholarly journals OR27-01 Combining Clinical and Genetic Approaches in Diagnosing a Large Brazilian Cohort of Patients with 46,XY Differences/Disorders of Sex Development (DSD)

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Nathalia Lisboa Rosa Almeida Gomes ◽  
Rafael Loch Batista ◽  
Mirian Yumie Nishi ◽  
Antonio Marcondes Lerario ◽  
Thatiana Evilen Silva ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
Massively Parallel Sequencing ◽  
Genetic Diagnosis ◽  
Disorders Of Sex Development ◽  
Androgen Insensitivity Syndrome ◽  
Unknown Etiology ◽  
Androgen Synthesis ◽  
Sex Development ◽  
Liquid Chromatography Tandem Mass ◽  
Phenotype Heterogeneity

Abstract Background: It is recommended a multidisciplinary approach consisted of clinical, hormonal and genetic workups for diagnosing 46,XY DSD. However, no previous study has quantified how useful is this combined approach. Objectives: To retrospectively review the clinical and genetic findings for diagnosing a large cohort of patients with 46,XY DSD from a single Brazilian center. Methods: 247 non-syndromic 46,XY DSD individuals (159 sporadic and 88 familial cases from 39 families) were studied. Clinical and hormonal data were collected from medical files. Testosterone (T), androstenedione (A) were measured by immunoradiometric or immunofluorimetric assays and dihydrotestosterone (DHT) by RIA after celite chromatography or by liquid chromatography tandem mass spectrometry; T/DHT and T/A ratios were calculated. Analysis of sensitivity (SE), specificity (SP) of T/DHT was performed, being the molecular diagnosis considered the gold standard for diagnosing SRD5A2 deficiency. A T/A>0.8 was considered indicative of 17ß-HSDB3 deficiency. The patients were clinically classified into four subgroups: 1) androgen insensitivity syndrome (AIS), 2) gonadal dysgenesis (GD); 3) defects in androgen synthesis (DAS) and 4) DSD of unknown etiology. Molecular studies were performed by Sanger sequencing and/ or massively parallel sequencing (MPS). Results: The median age at first visit was 14 years (range 0.1 to 59 years). The molecular diagnosis was established in 96.5% of the cases with AIS (n=28/29), in 96% of the subjects with DAS (n=46/48), in 36% of the patients with GD (n=21/57) and in 26.7% (n=15/56) with DSD of unknown etiology. The best cut-off for T/DHT in basal state and hCG stimulated was 12.5 (SE=100%; SP=78.57%) and 24 (SE=87.5%; SP=95.7%) respectively. A T/A<0.8 was observed in 13/16 (81%) of the patients with molecular diagnosis of 17ß-HSDB3 deficiency and also in 1/49 patients with other diagnose. Classification according to the phenotype matched with the genetic diagnosis in most cases. The molecular evaluation allowed that 16% (9/56) of the patients that were classified as DSD of unknow etiology had a definitive diagnosis, including six GD cases, two individuals with SRD5A2 deficiency and one with 17ß-HSDB3 deficiency. A clear AIS phenotype of five patients allowed us to consider and prove the pathogenicity of two synonymous and one promoter region variants as the cause of AIS. The combination of clinical and molecular diagnosis led to an increase in 8% the diagnosis in a total of 116 index-cases (58.5%) with a molecular diagnosis. Conclusion: Considering the phenotype heterogeneity, pitfalls of the hormonal assessment and number of genes involved, it is reasonable to consider MPS as a first test for diagnosing patients with 46,XY DSD. However, the combination of clinical and molecular diagnosis is more accurate than either strategies alone in diagnosing 46,XY DSD.

scholarly journals Outcomes of 4 years of molecular genetic diagnosis on a panel of genes involved in premature aging syndromes, including laminopathies and related disorders

2019 ◽  
Vol 14 (1) ◽  
Author(s):  
Maude Grelet ◽  
Véronique Blanck ◽  
Sabine Sigaudy ◽  
Nicole Philip ◽  
Fabienne Giuliano ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Diagnostic Yield ◽  
Genetic Diagnosis ◽  
Natural Aging ◽  
Premature Aging ◽  
Pathogenic Variants ◽  
Progeroid Syndromes ◽  
Premature Aging Syndromes

Abstract Background Segmental progeroid syndromes are a heterogeneous group of rare and often severe genetic disorders that have been studied since the twentieth century. These progeroid syndromes are defined as segmental because only some of the features observed during natural aging are accelerated. Methods Since 2015, the Molecular Genetics Laboratory in Marseille La Timone Hospital proposes molecular diagnosis of premature aging syndromes including laminopathies and related disorders upon NGS sequencing of a panel of 82 genes involved in these syndromes. We analyzed the results obtained in 4 years on 66 patients issued from France and abroad. Results Globally, pathogenic or likely pathogenic variants (ACMG class 5 or 4) were identified in about 1/4 of the cases; among these, 9 pathogenic variants were novel. On the other hand, the diagnostic yield of our panel was over 60% when the patients were addressed upon a nosologically specific clinical suspicion, excepted for connective tissue disorders, for which clinical diagnosis may be more challenging. Prenatal testing was proposed to 3 families. We additionally detected 16 variants of uncertain significance and reclassified 3 of them as benign upon segregation analysis in first degree relatives. Conclusions High throughput sequencing using the Laminopathies/ Premature Aging disorders panel allowed molecular diagnosis of rare disorders associated with premature aging features and genetic counseling for families, representing an interesting first-level analysis before whole genome sequencing may be proposed, as a future second step, by the National high throughput sequencing platforms (“Medicine France Genomics 2025” Plan), in families without molecular diagnosis.

scholarly journals An unusual presentation of X-linked adrenoleukodystrophy

2015 ◽  
Vol 2015 ◽  
Author(s):  
Avinash Suryawanshi ◽  
Timothy Middleton ◽  
Kirtan Ganda
Keyword(s):  
Blood Pressure ◽  
Genetic Testing ◽  
Adrenal Insufficiency ◽  
Spastic Paraparesis ◽  
Genetic Diagnosis ◽  
Sensory Loss ◽  
List Type ◽  
Luteinising Hormone Level ◽  
Childhood Onset ◽  
Primary Adrenal Insufficiency

Summary X-linked adrenoleukodystrophy (X-ALD) is a rare genetic condition caused by mutations in the ABCD1 gene that result in accumulation of very long chain fatty acids (VLCFAs) in various tissues. This leads to demyelination in the CNS and impaired steroidogenesis in the adrenal cortex and testes. A 57-year-old gentleman was referred for the assessment of bilateral gynaecomastia of 6 months duration. He had skin hyperpigmentation since 4 years of age and spastic paraparesis for the past 15 years. Physical examination findings included generalised hyperpigmentation (including skin, buccal mucosa and palmar creases), blood pressure of 90/60 mmHg, non-tender gynaecomastia and bilateral hypoplastic testes. Lower limb findings were those of a profoundly ataxic gait associated with significant paraparesis and sensory loss. Primary adrenal insufficiency was confirmed and investigations for gynaecomastia revealed normal testosterone with mildly elevated luteinising hormone level and normal prolactin. The combination of primary adrenal insufficiency (likely childhood onset), partial testicular failure (leading to gynaecomastia) and spastic paraparesis suggested X-ALD as a unifying diagnosis. A serum VLCFA panel was consistent with X-ALD. Subsequent genetic testing confirmed the diagnosis. Treatment with replacement doses of corticosteroid resulted in improvement in blood pressure and increased energy levels. We have reported the case of a 57-year-old man with a very late diagnosis of X-ALD manifested by childhood onset of primary adrenal insufficiency followed by paraparesis and primary hypogonadism in adulthood. Thus, X-ALD should be considered as a possibility in a patient with non-autoimmune primary adrenal insufficiency and neurological abnormalities. Learning points Adult patients with X-ALD may be misdiagnosed as having multiple sclerosis or idiopathic spastic paraparesis for many years before the correct diagnosis is identified. Screening for X-ALD with a VLCFA panel should be strongly considered in male children with primary adrenal insufficiency and in male adults presenting with non-autoimmune primary adrenal insufficiency. Confirmation of a genetic diagnosis of X-ALD can be very useful for a patient's family as genetic testing enables detection of pre-symptomatic female heterozygotes who can then be offered pre-natal testing to avoid transmission of the disease to male offsprings.

scholarly journals Genetic analysis of pediatric primary adrenal insufficiency of unknown etiology: 25 years’ experience in the UK

2021 ◽  
Author(s):  
Federica Buonocore ◽  
Avinaash Maharaj ◽  
Younus Qamar ◽  
Katrin Koehler ◽  
Jenifer P Suntharalingham ◽  
...  
Keyword(s):  
Young People ◽  
Genetic Analysis ◽  
Adrenal Insufficiency ◽  
Unknown Etiology ◽  
Candidate Gene Approach ◽  
Potential Candidate ◽  
Clinical Testing ◽  
Primary Adrenal Insufficiency ◽  
Children And Young People ◽  
Genetic Causes

Abstract Context Although primary adrenal insufficiency (PAI) in children and young people is often due to congenital adrenal hyperplasia (CAH) or autoimmunity, other genetic causes occur. The relative prevalence of these conditions is poorly understood. Objective We investigated genetic causes of PAI in children and young people over a 25 year period. Design, Setting and Participants Unpublished and published data were reviewed for 155 young people in the United Kingdom who underwent genetic analysis for PAI of unknown etiology in three major research centers between 1993-2018. We pre-excluded those with CAH, autoimmune or metabolic causes. We obtained additional data from NR0B1 (DAX-1) clinical testing centers. Intervention and Outcome Measurements Genetic analysis involved a candidate gene approach (1993 onwards) or next generation sequencing (NGS) (targeted panels, exomes) (2013-2018). Results A genetic diagnosis was reached in 103/155 (66.5%) individuals. In five children the adrenal insufficiency resolved and no genetic cause was found. Pathogenic variants occurred in 11 genes: MC2R (ACTH receptor) (30/155, 19.4%), NR0B1 (DAX-1) (7.7%), CYP11A1 (7.7%), AAAS (7.1%), NNT (6.5%), MRAP (4.5%), TXNRD2 (4.5%), STAR (3.9%), SAMD9 (3.2%), CDKN1C (1.3%) and NR5A1/SF-1 (0.6%). Additionally, 51 boys had NR0B1 variants identified through clinical testing. Although age at presentation, treatment, ancestral background and birthweight can provide diagnostic clues, genetic testing was often needed to define the cause. Conclusions PAI in children and young people often has a genetic basis. Establishing the specific etiology can influence management of this lifelong condition. NGS approaches improve the diagnostic yield when many potential candidate genes are involved.

Analysis of Sperm Motility Related to Transcriptional Alterations of Mitocondrial Genes in Males Affected by Infertility

2012 ◽  
Vol 10 (3) ◽  
pp. 455-462 ◽  
Author(s):  
V. Pietropaolo ◽  
C. Passariello ◽  
A. Bellizzi ◽  
A. Virga ◽  
E. Anzivino ◽  
...  
Keyword(s):  
Gene Expression ◽  
Sperm Motility ◽  
Expression Profiles ◽  
Mitochondrial Genes ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Control Group ◽  
Specific Gene ◽  
Over Expression ◽  
Significant Difference

Infertility is a problem afflicting about 1/6 couples, and in 40% of cases this is primarily due to the male. Male infertility is a multifactorial pathology and it seems mainly related to sperm motility or sperm number. However, a diagnosis of infertility is frequently not followed by a precise explanation of its cause, reflecting our poor understanding of the spermatogenesis-related regulatory mechanisms and gene expression profiles. Therefore, this study was design to investigate the relative gene expression of a specific gene profile in ejaculate spermatozoa of men affected by infertility. This profile included 13 mitochondrial gene encoding subunits of respiratory chain and 7 nuclear sperm motility-related genes. We used values of progressive sperm motility (PR) to separate subjects affected by infertility into two groups, showing PR values higher (H group) or lower (L group) than the mean of the sample, and to classify fertile men (control group). We did not obtain a statistically significant difference in nuclear gene expression patterns in spermatozoa among these three groups. On the other hand, we observed an over-expression in 11/13 tested mitochondrial genes in the population of infertile males with altered sperm motility compared to the control group. This over-expression led us to speculate that there is an abnormal mRNA transcription of these 11 subunits, that impaired the normal energy supply ensuring sperm motility. Regarding the under-expression of 2/13 tested mitochondrial genes, we could assume that the spermatozoa mtDNA has accumulated mutations involving these two genes (CYB and ND4L). In conclusion, our results will provide useful information for the development of molecular diagnostic tools for clinical assessment of sperm health. However, further investigation into other sperm-related genes is needed to establish their roles in male fertility.

The effects of Cyclosporin-A (CYA) on the Ultrastructure of Gingival Mast Cells (GMC) in Behcet's disease (BD)

1990 ◽  
Vol 48 (3) ◽  
pp. 358-359
Author(s):  
Oktay Arda ◽  
Ulkü Noyan ◽  
Selgçk Yilmaz ◽  
Mustafa Taşyürekli ◽  
İsmail Seçkin ◽  
...  
Keyword(s):  
Unknown Etiology ◽  
Control Group ◽  
Target Cells ◽  
Gingival Hyperplasia ◽  
Cellular Activity ◽  
Direct Relation ◽  
Immune Disease ◽  
Genital Ulceration ◽  
Functional Changes ◽  
The Third

Turkish dermatologist, H. Beheet described the disease as recurrent triad of iritis, oral aphthous lesions and genital ulceration. Auto immune disease is the recent focus on the unknown etiology which is still being discussed. Among the other immunosupressive drugs, CyA included in it's treatment newly. One of the important side effects of this drug is gingival hyperplasia which has a direct relation with the presence of teeth and periodontal tissue. We are interested in the ultrastructure of immunocompetent target cells that were affected by CyA in BD.Three groups arranged in each having 5 patients with BD. Control group was the first and didn’t have CyA treatment. Patients who had CyA, but didn’t show gingival hyperplasia assembled the second group. The ones displaying gingival hyperplasia following CyA therapy formed the third group. GMC of control group and their granules are shown in FIG. 1,2,3. GMC of the second group presented initiation of supplementary cellular activity and possible maturing functional changes with the signs of increased number of mitochondria and accumulation of numerous dense cored granules next to few normal ones, FIG. 4,5,6.

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