The clinical and molecular spectrum of the KDM6B-related neurodevelopmental disorder.

De novo variants are a leading cause of neurodevelopmental disorders (NDDs), but because every monogenic NDD is different and usually extremely rare, it remains a major challenge to understand the complete phenotype and genotype spectrum of any morbid gene. According to OMIM, heterozygous variants in KDM6B cause "neurodevelopmental disorder with coarse facies and mild distal skeletal abnormalities." Here, by examining the molecular and clinical spectrum of 85 reported individuals with mostly de novo (likely) pathogenic KDM6B variants, we demonstrate that this description is inaccurate and potentially misleading. Cognitive deficits are seen consistently in all individuals, but the overall phenotype is highly variable. Notably, coarse facies and distal skeletal anomalies, as defined by OMIM, are rare in this expanded cohort while other features are unexpectedly common (e.g., hypotonia, psychosis, etc.). Using 3D protein structure analysis and an innovative dual Drosophila gain-of-function assay, we demonstrated a disruptive effect of 11 missense/in-frame indels located in or near the enzymatic JmJC or Zn-containing domain of KDM6B. Consistent with the role of KDM6B in human cognition, we demonstrated a role for the Drosophila KDM6B ortholog in memory and behavior. Taken together, we accurately define the broad clinical spectrum of the KDM6B-related NDD, introduce an innovative functional testing paradigm for the assessment of KDM6B variants, and demonstrate a conserved role for KDM6B in cognition and behavior. Our study demonstrates the critical importance of international collaboration, sharing of clinical data, and rigorous functional analysis of genetic variants to ensure correct disease diagnosis for rare disorders.

SVEP1 as a Genetic Modifier of TEK-Related Primary Congenital Glaucoma.

Purpose: Affecting children by age 3, primary congenital glaucoma (PCG) can cause debilitating vision loss by the developmental impairment of aqueous drainage resulting in high intraocular pressure (IOP), globe enlargement, and optic neuropathy. TEK haploinsufficiency accounts for 5% of PCG in diverse populations, with low penetrance explained by variable dysgenesis of Schlemm's canal (SC) in mice. We report eight families with TEK-related PCG, and provide evidence for SVEP1 as a disease modifier in family 8 with a higher penetrance and severity. Methods: Exome sequencing identified coding/splice site variants with an allele frequency less than 0.0001 (gnomAD). TEK variant effects were assayed in construct-transfected HEK293 cells via detection of autophosphorylated (active) TEK protein. An enucleated eye from an affected member of family 8 was examined via histology. SVEP1 expression in developing outflow tissues was detected by immunofluorescent staining of 7-day mouse anterior segments. SVEP1 stimulation of TEK expression in human umbilical vascular endothelial cells (HUVECs) was measured by TaqMan quantitative PCR. Results: Heterozygous TEK loss-of-function alleles were identified in eight PCG families, with parent-child disease transmission observed in two pedigrees. Family 8 exhibited greater disease penetrance and severity, histology revealed absence of SC in one eye, and SVEP1:p.R997C was identified in four of the five affected individuals. During SC development, SVEP1 is secreted by surrounding tissues. SVEP1:p.R997C abrogates stimulation of TEK expression by HUVECs. Conclusions: We provide further evidence for PCG caused by TEK haploinsufficiency, affirm autosomal dominant inheritance in two pedigrees, and propose SVEP1 as a modifier of TEK expression during SC development, affecting disease penetrance and severity.

Genetic variants in ICAM1, PPARGC1A and MTHFR are potentially associated with different phenotypes of diabetic retinopathy.

PURPOSE: To explore phenotype-genotype correlations that may contribute to a better understanding of diabetic retinopathy (DR). PROCEDURES: An exploratory association study was performed to identify genetic variants associated with non-proliferative DR (NPDR) in 307 type 2 diabetic patients who were previously stratified into 3 different phenotypes of NPDR progression. The 307 patients were genotyped for 174 single nucleotide polymorphisms of 11 candidate genes (ACE, AGER, AKR1B1, ICAM1, MTHFR, NOS1, NOS3, PPARGC1A, TGFB1, TNF and VEGFA). RESULTS: Significant associations were observed for PPARGC1A rs16874120 with phenotype A (odds ratio, OR = 0.60, 95% confidence interval, CI 0.36-0.99), ICAM1 rs1801714 with phenotype B (OR = 3.32, 95% CI 1.05-10.50) and both PPARGC1A rs10213440 (OR = 2.00, 95% CI 1.07-3.73) and MTHFR rs1801133 (OR = 1.84, 95% CI 1.08-3.11) with phenotype C. CONCLUSIONS: RESULTS indicate that specific gene variants in ICAM1, PPARGC1A and MTHFR are associated with different NPDR phenotypes, being likely candidates to explain different disease mechanisms underlying the different phenotypes. This is the first study to show correlations between specific gene variants and NPDR phenotypes, opening new perspectives on DR.

Identification of DLEC1 D215N Somatic Mutation in Formalin Fixed Paraffin Embedded Melanoma and Melanocytic Nevi Specimens.

DLEC1 has been suggested as a tumor suppressor gene in several cancers. DLEC1 D215N somatic mutation (COSM36702) was identified in a melanoma cell line through whole genome sequencing. However, little is known about the implication and prevalence of this mutation in primary melanomas or in melanocytic nevi. The aim of this study was to genotype DLEC1 D215N mutation in melanoma tissue and melanocytic nevi samples to confirm its occurrence and to estimate its prevalence. Primary melanomas (n = 81) paired with synchronous or asynchronous metastases (n = 21) from 81 melanoma patients and melanocytic nevi (n = 28) were screened for DLEC1 D215N mutation. We found the mutation in 3 primary melanomas and in 2 melanocytic nevi, corresponding to a relatively low prevalence (3.7% and 7.1%, resp.). The pathogenic role of DLEC1 215N mutation is unclear. However, since the mutation has not been previously described in general population, its involvement in nevogenesis and melanoma progression remains a possibility to be clarified in future studies.

Assessment of the geographic origins of pinewood nematode isolates via single nucleotide polymorphism in effector genes.

The pinewood nematode, Bursaphelenchus xylophilus, is native to North America but it only causes damaging pine wilt disease in those regions of the world where it has been introduced. The accurate detection of the species and its dispersal routes are thus essential to define effective control measures. The main goals of this study were to analyse the genetic diversity among B. xylophilus isolates from different geographic locations and identify single nucleotide polymorphism (SNPs) markers for geographic origin, through a comparative transcriptomic approach. The transcriptomes of seven B. xylophilus isolates, from Continental Portugal (4), China (1), Japan (1) and USA (1), were sequenced in the next generation platform Roche 454. Analysis of effector gene transcripts revealed inter-isolate nucleotide diversity that was validated by Sanger sequencing in the genomic DNA of the seven isolates and eight additional isolates from different geographic locations: Madeira Island (2), China (1), USA (1), Japan (2) and South Korea (2). The analysis identified 136 polymorphic positions in 10 effector transcripts. Pairwise comparison of the 136 SNPs through Neighbor-Joining and the Maximum Likelihood methods and 5-mer frequency analysis with the alignment-independent bilinear multivariate modelling approach correlated the SNPs with the isolates geographic origin. Furthermore, the SNP analysis indicated a closer proximity of the Portuguese isolates to the Korean and Chinese isolates than to the Japanese or American isolates. Each geographic cluster carried exclusive alleles that can be used as SNP markers for B. xylophilus isolate identification.

Biometrical, Biochemical, and Molecular Diagnosis of Portuguese Meloidogyne hispanica Isolates.

Meloidogyne hispanica infects many economically important crops worldwide. The accurate identification of this pathogen is essential for the establishment of efficient and sustainable integrated pest management programs. Portuguese M. hispanica isolates were studied by biometrical, biochemical, and molecular characteristics. Biometrical characteristics of M. hispanica females, males, and second-stage juveniles were similar to the original description. Biochemical studies revealed a unique enzyme pattern (Hi4) for M. hispanica esterases that allowed for species differentiation. Molecular analysis of the mtDNA region from COII and 16S rRNA genes resulted in amplification products (1,800 bp) similar to M. hispanica, M. ethiopica, and M. javanica, and the described HinfI was unable to discriminate M. hispanica from the other two species. Analysis of the mtDNA sequences revealed altered nucleotides among the isolates that created new restriction sites for AluI and DraIII. The resulting restriction patterns successfully discriminated between the three species, providing a new tool for Meloidogyne identification. Finally, the phylogenetic relationship between M. hispanica and several Meloidogyne spp. sequences was analyzed using mtDNA, confirming the divergence between meiotic and mitotic species and revealing the proximity of M. hispanica to closely related species. Based on the studies conducted, the application of isozyme or polymerase chain reaction restriction fragment length polymorphism analysis would be a useful and efficient methodology for M. hispanica identification.

[Resection of bronchogenic cyst by video-mediastinoscopy]. / Ressecção de quisto broncogénico por vídeomediastinoscopia.

A bronchogenic cyst is a congenital malformation originating from the ventral primitive gut. It may be located in the mediastinum or in the lung parenchyma. Its location depends on the stage of gestation in which it developed. Despite being a histological benign tumor, many authors recommend its complete excision in order to obtain histological confirmation and to prevent future complications. The traditional approaches for excision are thoracotomy or video-assisted thoracoscopic surgery (VATS). However, a minimally invasive approach through vídeomediastinoscopy constitutes a valid alternative in selected cases. The authors present a case of a 23 years old female patient, admitted to the emergency department with retrosternal pain, dyspnea at rest, tachycardia, polypnea, infra-clavicular accessory muscle contraction, jugular vein distention and hypoxemia. A Chest CT revealed a large cystic lesion of the middle mediastinum, with compression of the vascular structures, deviation of the tracheobronchial tree and reduction in the diameter of the main bronchi. The patient was referred for surgical treatment. A mini-cervicotomy incision was made, and with elevation of the sternum a video-mediastinoscope was introduced. With bimanual instrumentation, complete excision of a large mediastinal cyst of the middle mediastinum was performed. The cyst was located in the subcarinal and pre-tracheal space, had contact with the left and right main bronchi, esophagus, roof of the left atrium, pulmonary artery and superior pulmonary veins. The histological evaluation revealed a bronchogenic cyst. There were no immediate postoperative complications. The patient remains asymptomatic after two years of follow-up, and without recurrence.

A rare case of co-infection with pulmonary tuberculosis and oronasal actinomycosis.

Oronasal actinomycosis is an infection seldom described in the literature, especially in the form of a co-infection with pulmonary tuberculosis. We report the case of a 48-year-old male admitted to the isolation ward due to active pulmonary tuberculosis, with a history of diabetes and alcohol abuse. While hospitalized, the patient complained of dysphagia and nasal regurgitation of food. The examination of the oral cavity revealed an oronasal fistula. The infecting agent was identified, and the treatment was successful. We also present a brief review of the literature, as well as a full description and discussion of the process of investigating this rare clinical case.

A rare case of co-infection with pulmonary tuberculosis and oronasal actinomycosis / Caso raro de coinfecção tuberculose pulmonar e actinomicose oronasal

Oronasal actinomycosis is an infection seldom described in the literature, especially in the form of a co-infection with pulmonary tuberculosis. We report the case of a 48-year-old male admitted to the isolation ward due to active pulmonary tuberculosis, with a history of diabetes and alcohol abuse. While hospitalized, the patient complained of dysphagia and nasal regurgitation of food. The examination of the oral cavity revealed an oronasal fistula. The infecting agent was identified, and the treatment was successful. We also present a brief review of the literature, as well as a full description and discussion of the process of investigating this rare clinical case.

A actinomicose oronasal é uma infecção raramente descrita na literatura, especialmente na forma de coinfecção com tuberculose pulmonar. Descrevemos o caso de um paciente de 48 anos de idade, admitido em enfermaria de isolamento por tuberculose pulmonar bacilífera, com história de diabetes e alcoolismo. Durante a internação, o paciente referiu queixas de disfagia e regurgitação alimentar por via nasal. O exame da cavidade oral revelou uma fístula oronasal. O agente infeccioso foi identificado, e o tratamento foi realizado com sucesso. Apresentamos também uma breve revisão da literatura e uma descrição e discussão completa do processo de investigação deste raro caso clínico.

Lung adenocarcinoma - For how long a good response is the question. / Adenocarcinoma do pulmão - Até quando a boa resposta?

A 75 year old woman was observed at the emergency department with cough, hyperthermia and thoracic pain in October 2004.A pleural effusion was identified and studied. Thorax CT scan evidenced a pleural effusion and thickening in the RSL anterior segment but bronchofibroscopy only identified inflammatory changes. In February 2005 the pleural effusion relapsed and the CT scan showed nodular densification adjacent to LSLB and lingual. VATS converted to thoracotomy was preformed and followed by a long recover period. Histology of surgical specimen revealed an adenocarcinoma. First line CT was administered until progression as bone metastasis, in March 2006. Second line CT was started, with disease stabilization. The administration of erlotinib was started in July 2007 and continues to the present with good tolerance. The patient has no pain and continues without progression of disease after 13 month of treatment. For how long will she maintain this response, is the question. Rev Port Pneumol 2008; XIV (Supl 3): S71-S77.