Displasia ectodérmica hipohidrótica ligada al cromosoma X de novo por variante recurrente en un paciente mexicano / X-linked hypohidrotic ectodermal dysplasia by a de novo recurrent variant in a Mexican patient

Resumen Introducción: Las displasias ectodérmicas son un grupo de genodermatosis que se caracterizan por distrofia de las estructuras derivadas del ectodermo. De ellas, la variedad más común es la hipohidrótica, con una incidencia de 7/100,000 nacidos vivos observada en todos los grupos étnicos. La displasia ectodérmica hipohidrótica tiene distintas etiologías. La presentación más frecuente es la asociada a un patrón de herencia ligado al cromosoma X, causada por variantes patogénicas del gen EDA en Xq13.1. EDA codifica a la ectodisplasina A, una molécula de señalización que participa en la comunicación epitelio-mesénquima durante el desarrollo de la piel y los anexos. Caso clínico: Varón de 6 años con las características clínicas cardinales de la displasia ectodérmica hipohidrótica ligada al cromosoma X (DEHLX), que incluyen hipotricosis, oligodoncia e hipohidrosis. El análisis del gen EDA por secuenciación directa mostró la presencia de la variante patogénica c.466C>T, p.Arg156Cys, rs132630313 con presentación de novo en el paciente. Esta variante ya ha sido reportada en diferentes poblaciones, incluyendo familias mexicanas, y constituye un punto caliente para mutación en EDA. Se analizaron los hallazgos clínicos, la etiología y el manejo de la DEHLX, en la que de manera reciente se ha planteado la posibilidad de otorgar tratamiento prenatal para prevenir sus manifestaciones clínicas. Conclusiones: Se pone de relevancia que el análisis molecular en pacientes con DEHLX corrobora el diagnóstico clínico y permite brindar asesoramiento genético con bases moleculares.

Abstract Background: Ectodermal dysplasias are a group of genodermatoses characterized by dystrophy of ectodermal derived structures. The most frequent presentation of the ectodermal dysplasias is the hypohidrotic type, which has an incidence of 7/100,000 newborns and has been described in all ethnic groups. The hypohidrotic ectodermal dysplasia (HED) has different etiologies, and it is more frequently associated with an X-linked pattern of inheritance caused by pathogenic variants of the EDA gene in Xq13.1. EDA encodes the protein ectodisplasin A, a signal molecule which participates in epithelium and mesenchymal development of the skin. Case report: A 6 year-old male patient with the main clinical characteristics of the X-linked HED including hypotrichosis, hypodontia and hypohidrosis. The direct sequencing analysis of EDA in our patient detected a de novo pathogenic variant, c.466C>T, p.Arg156Cys, rs132630313. This variant has been previously described in different ethnic groups, including Mexican families, and is considered a mutational hotspot. The clinical characteristics, etiology and management of the X-linked HED, including the possibility of prenatal therapy in order to avoid the clinical manifestations are discussed. Conclusions: The molecular analysis in patients with X-linked HED is of relevance, as it enables to confirm the clinical diagnosis and also, it allows a genetic assessment with molecular bases.

Detection of EDA gene mutation and phenotypic analysis in patients with hypohidrotic ectodermal dysplasia / 北京大学学报(医学版)

OBJECTIVE@#To detect the ectodysplasin A (EDA) gene mutation in patients with hypohidro-tic ectodermal dysplasia (HED), and to analyze the distribution pattern of missing permanent teeth and the systemic manifestation of HED patients with EDA gene mutation.@*METHODS@#Twelve HED families were enrolled from clinic for genetic history collection, systemic physical examination and oral examination. Peripheral blood or saliva samples were collected from the probands and the family members to extract genomic DNA. PCR amplification and Sanger sequencing were utilized to detect the EDA gene variations, which were compared with the normal sequence (NM_001399.5). The functional impact of EDA gene variants was then evaluated by functional prediction of mutation, conservation analysis and protein structure prediction. The pathogenicity of each EDA gene variation was assessed according to the stan-dards and guidelines of the American College of Medical Genetics and Genomics (ACMG). The systemic phenotype and missing permanent tooth sites of HED patients with EDA gene mutations were summarized, and the missing rate of each tooth position was analyzed and compared.@*RESULTS@#Eight out of twelve HED families were identified to carry EDA gene mutations, including: c.164T>C(p.Leu55Pro); c.457C>T (p.Arg153Cys); c.466C>T(p.Arg156Cys); c. 584G>A(p.Gly195Glu); c.619delG(p.Gly207Profs*73); c.673C>T(p.Pro225Ser); c.676C>T(p.Gln226*) and c.905T>G(p.Phe302Cys). Among them, c.164T>C(p.Leu55Pro); c.619delG(p.Gly207Profs*73); c.673C>T(p.Pro225Ser); c.676C>T(p.Gln226*) and c.905T>G(p.Phe302Cys) were novel mutations. The HED patients with EDA gene mutations in this study were all male. Our results showed that the average number of missing permanent teeth was 13.86±4.49, the average number of missing permanent teeth in the upper jaw was 13.14±5.76, the missing rate was 73.02%. And in the lower jaw, the average number of missing permanent teeth was 14.57±3.05, the missing rate was 80.95%. There was no significant difference in the number of missing teeth between the left and right sides of the permanent dentition (P>0.05). Specifi-cally, the maxillary lateral incisors, the maxillary second premolars and the mandibular lateral incisors were more likely to be missing, while the maxillary central incisors, the maxillary and mandibular first molars had higher possibility of persistence.@*CONCLUSION@#This study detected novel EDA gene pathogenic variants and summarized the distribution pattern of missing permanent teeth of HED patients, thus enriched the variation and phenotype spectrum of EDA gene, and provided new clinical evidence for genetic diagnosis and prenatal consultation.

Association Of Eda Gene Polymorphisms Rs 372024, Rs 3764746, Rs 3795170 In Skeletal Class I Cases With Dental Crowding

The EDA gene, associated with X linked hydrotic form of Ectodermal Dysplasia, its mutations could potentially lead to differential gene expression that causes large tooth phenotype, which has been suggested to cause dental crowding. We analyzed the association of genetic polymorphisms in EDA gene variants rs 372024, rs 3764746, and rs 3795170 among Skeletal Class I crowding cases using blood samples of 30 cases and 30 controls, which were subjected to PCR amplication and DNA sequencing. Based on the statistical analysis using the Z test we found CG and GG genotype for rs3764746 and GT and TT genotype for rs3795170 showed a statistically signicant result. These results suggest that EDA gene variants rs3764746 and rs3795170 could be genetic markers for dental crowding in our population while EDA gene variant rs372024 did not show any signicant association in our population. These ndings can provide in-depth knowledge, regarding the genetic inuences on the incidence of crowding of teeth.

Effects of negative pressure wound therapy on the expression of EDA+FN in granulation tissues of human diabetic foot wounds / 解放军医学杂志

Objective To investigate the effects of negative pressure wound therapy (NPWT) on the expression of EDA+ FN in granulation tissues of human diabetic foot wounds.Methods Forty patients with diabetic foot wounds fitting the inclusion criteria,admitted from Jan.2014 to Jun.2016,were randomly and equally apportioned to receive either NPWT or conventional gauze therapy (control) for 14 days.Granulated tissue biopsies were collected before (0 day) and after (14 day) treatment in both groups.All biopsies were subdivided into two parts.One part was preserved in 4％ paraformaldehyde for immunocytochemical staining of EDA+FN,and the other part was stored at-80 ℃ for Western blotting and PCR analysis of EDA+FN.Results The immunohistochemical analysis revealed that the mean area density of EDA+ FN increased in both NPWT group and control group at day 14 relative to day 0,but the change value of mean area density was higher in NPWT group than in control group (P＜0.01).Western blotting showed that the relative protein levels of EDA+FN increased in both NPWT group and control group at day 14 relative to day 0,but the change value of relative protein levels of EDA+FN was higher in NPWT group than in control group (P＜0.01).The real time PCR analysis demonstrated that the relative mRNA levels of EDA+ FN increased in both NPWT group and control group at day 14 relative to day 0,but the change value of relative mRNA levels of EDA+ FN was higher in NPWT group than in control group (P＜0.01).The results demonstrated the higher protein and mRNA levels of EDA+FN in NPWT group than that in control group.Conclusion NPWT obviously enhances EDA+FN expression in granulation tissue of diabetic foot wound,as a result promotes wound healing.

EDA mutation screening and phenotype analysis in patients with tooth agenesis / 北京大学学报(医学版)

Objective:To screen the ectodysplasin A (EDA)gene mutation in the patients with non-syndromic tooth agenesis and ectodermal dysplasia,and to analyze the phenotype of missing teeth pattern in these two groups of patients.Methods:In the study,174 patients with tooth agenesis (143:non-syn-dromic,31:ectodermal dysplasia)and 451 health control volunteers were enrolled from the clinic,and the genome DNA was extracted from either peripheral blood or oral mucosal swab.The coding region of EDA gene was then amplified by PCR,sequenced and blasted to online NCBI database.The missing teeth were recorded for all patients,and the missing teeth from patients with EDA mutation were com-pared among the different dentition sites.Results:33 patients were identified with EDA mutation.In the non-syndromic patients,13 /143(9.09%)were identified with EDA mutation,while in patients with ec-todermal dysplasia,20 /31 (64.52%)were found with EDA mutation.Ten novel EDA mutations were identified (c.769G >C[p.G257R],c.936C >G[p.I312M],c.223G >A[p.E75K],c.1166C >T[p. P389L],c.133G >C[p.G45R],c.1109G >A[p.E370K],c.914G >T[p.S305I],c.916C >T[p. Q306X],c.602G >T[p.G201V],c.88 -89insG[p.A30GfsX69]).For each dentition site there was no statistic difference in the number of missing teeth between the left and right sides,so the number from both sides were combined later in the analysis.In the patients with EDA mutation,the non-syndromic pa-tients had fewer missing teeth (15.9 ±6.4 missing teeth for each,207 /364 in total)than the patients with ectodermal dysplasia (23.9 ±4.3,478 /560).In the non-syndromic patients with EDA mutation, the maxillay central incisors and first molars were less affected,with the same missing rate as 19.2% (5 /26).While the mandibular central incisors (with a missing rate of 76.9%,20 /26),the maxillary late-ral incisors (the missing rate:88.5%,23 /26 ),the mandibular lateral incisors (the missing rate:80.8%,21 /26),and the maxillary first premolars (the missing rate:80.8%,21 /26)were more likely to be missing.In the ectodermal dysplasia patients with EDA mutation,only maxillary central incisors (the missing rate:60%,24 /40),maxillary canines (the missing rate:70%,28 /40),mandibular ca-nines (the missing rate:67.5%,27 /40),maxillary first molars (the missing rate:65%,26 /40)and mandibular first molars (the missing rate:72.5%,29 /40)had higher possibility of persistence.Teeth at other dentition sites were more likely to be affected (the minimum missing rate:87.5%,35 /40). Conclusion:The findings would help to reveal the EDA gene and its function in ectodermal organogene-sis.

Estudio clínico y molecular en un escolar con displasia ectodérmica hipohidrótica ligada al X / Clinical and molecular study in a child with X-linked hypohidrotic ectodermal dysplasia

Las displasias ectodérmicas comprenden más de 200 entidades clínicamente distintivas, las cuales afectan, al menos, dos estructuras derivadas del ectodermo, que incluyen la piel, el pelo, las unas, los dientes, las glándulas sudoríparas y sebáceas. La displasia ectodérmica hipohidrótica ligada al X es el tipo más frecuente y es causada por mutación del gen EDA, que codifica la ectodisplasina-A. Su frecuencia es menor de 1 en 100000 individuos y se caracteriza clínicamente por presentar hipodoncia, hipohidrosis, hipotricosis y alteraciones oculares. Se expone el caso de un escolar evaluado de forma multidisciplinaria con diagnóstico clínico y molecular de displasia ectodérmica hipohidrótica ligada al X con mutación tipo cambio de sentido c.1133C>,T, p.T378M, en el gen EDA.

Ectodermal dysplasia encompasses more than 200 clinically distinct entities, which affect at least two structures derived from the ectoderm, including the skin, hair, nails, teeth, sweat glands, and sebaceous glands. X-linked hypohidrotic ectodermal dysplasia is the most common type and is caused by mutation of the EDA gene that encodes Ectodysplasin-A. It occurs in less than 1 in 100 000 individuals and is clinically characterized by hypodontia, hypohidrosis, hypotrichosis, and eye dis orders. We present a child evaluated in a multidisciplinary manner with clinical and molecular diagnosis of X-linked hypohidrotic ectodermal dysplasia with type missense mutation c.1133C> T; p.T378M in EDA gene.

Molecular diagnosis of X-linked hypohidrotic ectodermal dysplasia families / 中华实用儿科临床杂志

Objective To explore the mutations of EDA gene in 2 X - linked hypohidrotic ectodermal dyspla-sia(XLHED)pedigrees,and provide clues for the XLHED diagnosis,genetic counseling and treatment. Methods Polymerase chain reaction and direct sequencing were used to analyze the coding sequences and their flanking sequences of the EDA gene in the patients,suspicious carriers,normal family members in 2 families and non - relative control sam-ples. Results In family 1,mutation c. 659 676del18,namely p. 220 225del(Gly - X - Y)6 which was located in (Gly - X - Y)19 collagen - like repeat domain,was found in the proband and other patient's EDA gene. In family 2,an insertion c. 118 - 119insT was found in the intracellular domain,which induces reading frame alteration from the 40th a-mino acid. The mutations found in the 2 families were consistent with the principle of mutation and phenotype co - sepa-ration,but these mutations were not found in the normal control samples. EDA gene analysis of fetal amniotic fluid sam-ple from Ⅲ - 1 in the family 1 was not found to have the same mutation as the proband,and the follow - up after birth proved normal for the baby. Conclusions EDA gene c. 118 - 119insT mutation found in the research is a novel muta-tion. Sequence analysis of EDA gene is an efficient method in XLHED diagnosis,and is beneficial for the genetic coun-seling and the genetic intervention of the disease in the affected families.

Enfermedad diarreica aguda por Escherichia coli enteropatógenas en Colombia / Acute diarrheal disease caused by enteropathogenic Escherichia coli in Colombia

Intestinal Escherichia coli pathogens are leading causes of acute diarrheal disease in children less than 5 years in Latin America, Africa and Asia and a leading cause of death in children living in poorest communities in Africa and South East Asia. Studies on the role of E. coli pathogens in childhood diarrhea in Colombia and other countries in Latin America are limited due to the lack of detection assays in clinical laboratories at the main urban medical centers. Recent studies report that enterotoxigenic E. coli is the most common E. coli pathogens associated with diarrhea in children less than 5 years of age. Other E. coli pathotypes have been detected in children with diarrhea including enteropathogenic, enteroaggregative, shiga-toxin producing and diffusely adherent E. coli. It was also found that meat and vegetables at retail stores are contaminated with Shiga-toxin producing E. coli and enteroaggregative E. coli, suggesting that food products are involved in transmission and infection of the susceptible host. More studies are necessary to evaluate the mechanisms of transmission, the impact on the epidemiology of diarrheal disease, and management strategies and prevention of these pathogens affecting the pediatric population in Colombia.

Las cepas de E. coli enteropatógenas son causas importantes de la enfermedad diarreica aguda (EDA) en niños bajo 5 años de edad en América Latina, África y Asia y están asociadas a alta mortalidad en niños en las comunidades más pobres de África y el Sudeste Asiático. Estudios sobre el papel de las variedades de E. coli entero-patógenas en la EDA infantil en Colombia y otros países de América Latina son limitados debido a la carencia de ensayos para detección de estos patógenos en los laboratorios clínicos de centros de salud. Estudios recientes han reportado la detección de E. coli enteropatógenas en Colombia, siendo E. coli enterotoxigénica la cepa más frecuentemente asociada a diarrea en niños bajo 5 años. Otros patógenos detectados en estos pacientes incluyen E. coli enteroagregativa, enteropatógena "clásica", productora de toxina Shiga, y de adherencia difusa. Con base en estudios que reportan la presencia de E. coli productora de toxina Shiga y E. coli enteroagregativa en carnes y vegetales en supermercados, se cree que productos alimentarios contaminados contribuyen a la transmisión de estos patógenos y a la infección del hospedero susceptible. Más estudios son necesarios para evaluar los mecanismos de transmisión, el impacto en la epidemiologia de la EDA, y las pautas de manejo y prevención de estos patógenos que afectan la población pediátrica en Colombia.

Effect of EDA-A1 gene mutant on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell ECV304 / 重庆医学

Objective To investigate the effect of ectodysplasin A (EDA‐A1) gene of hypohidrotic ectodermal dysplasia on pro‐liferation and cell cycle of human umbilical vein endothelial cell (ECV304). Methods Recombinant eukaryotic expression vectors pcDNA3. 1(‐)‐EDA‐A1‐M /W (mutant, M and wild, W) containing the coding sequence were transected into ECV304 cells. EDA‐A1 gene was amplified by reverse transcription polymerase chain reaction (RT‐PCR), and the protein was detected by Western blot. Cell viability and cycle distribution were invested by MTT and Flow cytometry (FCM ). Results The EDA‐A1 gene and pro‐tein were detected respectively by RT‐PCR and Western blot in ECV cells transfected with pcDNA3. 1(‐)‐EDA‐A1‐M /W, but not in ECV cells transfected with plasmid pcDNA3. 1(‐) and cells without transection. And also, compared with control groups, EDA‐A1 gene mutant significantly decreased proliferation of ECV cells and its inhibition rate was 45. 70% ( P 0. 05). A significant increase of the G0 /G1 and S fraction was seen in the ECV cells of mutant group, compared with wild group with an accumulation in S phase and a concomitant decrease in G2 /M phase population (P< 0. 05). Conclusion Mutant and wild EDA‐A1 gene may have distinct biological functions on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell.

Incorporación del componente investigativo para la enseñanza de la enfermedad de diarrea aguda en la formación del médico general / Inclusion of the investigative component to learn about acute diarrheic disease during the training of medical doctors

La morbilidad por las enfermedades diarreicas agudas (EDA) sigue constituyendo un problema de salud, a pesar de que se han logrado avances en la reducción de las cifras de mortalidad. Se realizó una investigación aplicada, descriptiva, explicativa, longitudinal prospectiva, con el objetivo de caracterizar los principales factores de riesgo socio ambientales de las EDA en el área de salud, mediante la vinculación del estudiante de medicina con la familia del niño ingresado en el Hospital Pediátrico "Pepe Portilla" (año 2007-2008). El universo estuvo constituido por 731 niños, con un muestreo aleatorio simple de 252. Se utilizaron los métodos de la encuesta, análisis documental, observación mediante la visita a los hogares. Se procesaron los resultados mediante la estadística descriptiva y arribaron a los siguientes resultados: el estado nutricional detectado fue de 66 niños con afectación del estado nutricional y 186 sin afectación, vinculándose a los primeros las peores condiciones higiénicas sanitarias. En ambos grupos se detectaron deficiencias en las acciones de salud que deben ser instrumentadas en la atención primaria sobre prevención-promoción de las EDA y el seguimiento nutricional a lactantes; por lo que se realizaron charlas educativas por los estudiantes de Medicina sobre las orientaciones higiénicas epidemiológicas en la prevención de la diarrea. Se logró incorporar el componente investigativo en el proceso docente educativo de la formación del estudiante de Medicina.

The morbidity due to Acute Diarrheic Disease (ADD) still constitutes a health problem, though the figures of mortality have been reduced. An applied, descriptive, explanatory, longitudinal-prospective research was conducted with the purpose of characterizing the main socio-environmental risk factors of the ADD in the health area, linking the medical students with the family of the child admitted at "Pepe Portilla" Children Provincial Hospital (2007-2008). The target group was comprised of 731 children with a sample of 252 taken at random. The methods used were the survey, the documentary analysis and the observation through home visits. The results were processed using the descriptive statistics with the following results: 66 children with poor nutritional status in close relation to the worst hygienic conditions and 186 children without affectation. Deficiencies were detected in both groups when taking the health actions to be implemented in Primary Health Care regarding the prevention of ADD, health promotion work and a follow up of the nutritional status of the infants; that was why medical students organized educational talks about epidemiological-hygienic orientations to prevent the ADD. The investigative component about this topic was included in the teaching-learning process to train medical students.

Gene mutation detection for a patient with anhidrotic ectodermal dysplasia / 实用口腔医学杂志

Objetive:To detect the gene mutation of a patient with anhidrotic ectodermal dysplasia. Methods: Genomic DNA was extracted from the peripheral blood of the patient and all the exon fragments were obtained by PCR. These fragments were sequenced directly. Results: In EDA gene a novel base transition was detected: -48A→G. Conclusion: There is a new base transition in EDA gene in this patient

The Effect of Emotionality in Lie- detection Questions on Skin Conductance Response / 中国临床心理学杂志

Objective: to examine the relationship between emotional states and skin conductance response during lie- detection and to provide evidence for improvement of lie- detection technique. Methods: 38 university students participat- ed in the study. They were provided with antisocial behavior materials that elicited different levels of emotional arousal. Results: Significant difference in skin conductance response was observed with the high emotionality items eliciting the highest skin conductance response. Conclusion: Under a certain level of pressure, individuals’emotional state has direct impact on skin conductance. High emotionality questions are better for detecting lies and honesty than low emotionality questions. In addition to lying, the questions themselves have direct impact on subjects’emotional responses, which in turn lead to skin conductance response. When cues for emotionality are obvious, related events tend to receive greater at- tention by subjects and thus lead to special skin conductance response.