PRPH2-Related Retinal Dystrophies: Mutational Spectrum in 103 Families from a Spanish Cohort.

PRPH2, one of the most frequently inherited retinal dystrophy (IRD)-causing genes, implies a high phenotypic variability. This study aims to analyze the PRPH2 mutational spectrum in one of the largest cohorts worldwide, and to describe novel pathogenic variants and genotype-phenotype correlations. A study of 220 patients from 103 families recruited from a database of 5000 families. A molecular diagnosis was performed using classical molecular approaches and next-generation sequencing. Common haplotypes were ascertained by analyzing single-nucleotide polymorphisms. We identified 56 variants, including 11 novel variants. Most of them were missense variants (64%) and were located in the D2-loop protein domain (77%). The most frequently occurring variants were p.Gly167Ser, p.Gly208Asp and p.Pro221_Cys222del. Haplotype analysis revealed a shared region in families carrying p.Leu41Pro or p.Pro221_Cys222del. Patients with retinitis pigmentosa presented an earlier disease onset. We describe the largest cohort of IRD families associated with PRPH2 from a single center. Most variants were located in the D2-loop domain, highlighting its importance in interacting with other proteins. Our work suggests a likely founder effect for the variants p.Leu41Pro and p.Pro221_Cys222del in our Spanish cohort. Phenotypes with a primary rod alteration presented more severe affectation. Finally, the high phenotypic variability in PRPH2 hinders the possibility of drawing genotype-phenotype correlations.

RPE65-related retinal dystrophy: Mutational and phenotypic spectrum in 45 affected patients.

INTRODUCTION: Biallelic pathogenic RPE65 variants are related to a spectrum of clinically overlapping inherited retinal dystrophies (IRD). Most affected individuals progress to severe disease, with 50% of patients becoming legally blind by 20 years of age. Deeper knowledge of the mutational spectrum and the phenotype-genotype correlation in RPE65-related IRD is needed. PATIENTS AND METHODS: Forty-five affected subjects from 27 unrelated families with a clinical diagnosis of RPE65-related IRD were included. Clinical evaluation consisted of self-reported ophthalmological history and objective ophthalmological examination. Patients' genotype was classified according to variant class (truncating or missense) or to variant location at different protein domains. The main phenotypic outcome measure was age at onset (AAO) of symptomatic disease and a Kaplan-Meier analysis of disease symptom event-free survival was performed. RESULTS: Twenty-nine different RPE65 variants were identified in our cohort, 7 of them novel. Patients carrying two missense alleles showed a later disease onset than those with 1 or 2 truncating variants (log-rank test p <0.05). While 60% of patients carrying a missense/missense genotype presented symptoms before or during the first year of life, almost all patients with at least 1 truncating allele (91%) had an AAO ≤1 year (p <0.05). CONCLUSION: Our findings suggest an association between the type of RPE65 variant carried and AAO. These findings provide useful data on RPE65-associated IRD phenotypes and may help improve clinical and therapeutic management of these patients.

Exome sequencing identifies PEX6 mutations in three cases diagnosed with Retinitis Pigmentosa and hearing impairment.

Purpose: The aim of the present work is the molecular diagnosis of three patients with deafness and retinal degeneration. Methods: Three patients from two unrelated families were initially analyzed with custom gene panels for Usher genes, non-syndromic hearing loss, or inherited syndromic retinopathies and further investigated by means of clinical or whole exome sequencing. Results: The study allowed us to detect likely pathogenic variants in PEX6, a gene typically involved in peroxisomal biogenesis disorders (PBDs). Beside deaf-blindness, both families showed additional features: Siblings from Family 1 showed enamel alteration and abnormal peroxisome. In addition, the brother had mild neurodevelopmental delay and nephrolithiasis. The case II:1 from Family 2 showed intellectual disability, enamel alteration, and dysmorphism. Conclusions: We have reported three new cases with pathogenic variants in PEX6 presenting with milder forms of the Zellweger spectrum disorders (ZSD). The three cases showed distinct clinical features. Thus, expanding the phenotypic spectrum of PBDs and ascertaining exome sequencing is an effective strategy for an accurate diagnosis of clinically overlapping and genetically heterogeneous disorders such as deafness-blindness association.

Genomic Landscape of Sporadic Retinitis Pigmentosa: Findings from 877 Spanish Cases.

PURPOSE: We aimed to unravel the molecular basis of sporadic retinitis pigmentosa (sRP) in the largest cohort reported to date. DESIGN: Case series. PARTICIPANTS: A cohort of 877 unrelated Spanish sporadic cases with a clinical diagnosis of retinitis pigmentosa (RP) and negative family history. METHODS: The cohort was studied by classic genotyping or targeted next-generation sequencing (NGS). Multiplex ligation-dependent probe amplification (MLPA) and array-based comparative genomic hybridization were performed to confirm copy number variations detected by NGS. Quantitative fluorescent polymerase chain reaction was assessed in sRP cases carrying de novo variants to confirm paternity. MAIN OUTCOME MEASURES: The study of the sRP cohort showed a high proportion of causal autosomal dominant (AD) and X-linked (XL) variants, most of them being de novo. RESULTS: Causative variants were identified in 38% of the patients studied, segregating recessively in 84.5% of the solved cases. Biallelic variants detected in only 6 different autosomal recessive genes explained 50% of the cases characterized. Causal AD and XL variants were found in 7.6% and 7.9% of cases, respectively. Remarkably, 20 de novo variants were confirmed after trio analysis, explaining 6% of the cases. In addition, 17% of the solved sRP cases were reclassified to a different retinopathy phenotype. CONCLUSIONS: This study highlights the clinical utility of NGS testing for sRP cases, expands the mutational spectrum, and provides accurate prevalence of mutated genes. Our findings evidence the underestimated role of de novo variants in the etiology of RP, emphasizing the importance of segregation analysis as well as comprehensive screening of genes carrying XL and AD variants in sporadic cases. Such in-depth study is essential for accurate family counseling and future enrollment in gene therapy-based treatments.

[Family structure of obese/overweight and normal weight adolescents in México]. / Estructura familiar y estado de nutrición en adolescentes de Sonora, México.

BACKGROUND: Feeding modes and appetence toward certain foods are usually conditioned by the family. Obesity during adolescence usually persists during adulthood. AIM: To determine differences in family structure of adolescents according to their nutritional status. MATERIAL AND METHODS: A cross-sectional study was conducted in 60 overweight-obese and 60 normal weight adolescents. Family type was determined based on their conformation (kinship and cohabitation), development (if the mother had a remunerated job), demography (geographical area), integration (functions of the couple); life cycle stage and functionality. RESULTS: Fifty eight percent of normal weight adolescents had simple nuclear families and 47% of overweight-obese adolescents had an extended family. Thirty one and 21% of overweight/obese and normal weight adolescents lived with an overweight/obese individual, respectively. CONCLUSIONS: There are differences in the family structure of overweight/obese and normal weight adolescents.

Comprehensive Genotyping and Phenotyping Analysis of GUCY2D-Associated Rod- and Cone-Dominated Dystrophies.

PURPOSE: To describe the genetic and clinical spectrum of GUCY2D-associated retinopathies and to accurately establish their prevalence in a large cohort of patients. DESIGN: Retrospective case series. METHODS: Institutional study of 47 patients from 27 unrelated families with retinal dystrophies carrying disease-causing GUCY2D variants from the Fundación Jiménez Díaz hospital dataset of 8000 patients. Patients underwent ophthalmological examination and molecular testing by Sanger or exome sequencing approaches. Statistical and principal component analyses were performed to determine genotype-phenotype correlations. RESULTS: Four clinically different associated phenotypes were identified: 66.7% of families with cone/cone-rod dystrophy, 22.2% with Leber congenital amaurosis, 7.4% with early-onset retinitis pigmentosa, and 3.7% with congenital night blindness. Twenty-three disease-causing GUCY2D variants were identified, including 6 novel variants. Biallelic variants accounted for 28% of patients, whereas most carried dominant alleles associated with cone/cone-rod dystrophy. The disease onset had statistically significant differences according to the functional variant effect. Patients carrying GUCY2D variants were projected into 3 subgroups by allelic combination, disease onset, and presence of nystagmus or night blindness. In contrast to patients with the most severe phenotype of Leber congenital amaurosis, 7 patients with biallelic GUCY2D had a later and milder rod form with night blindness in infancy as the first symptom. CONCLUSIONS: This study represents the largest GUCY2D cohort in which 4 distinctly different phenotypes were identified, including rare intermediate presentations of rod-dominated retinopathies. We established that GUCY2D is linked to about 1% of approximately 3000 molecularly characterized families of our cohort. All of these findings are critical for defining cohorts for inclusion in future clinical trials.

Are Mexican agricultural farmlands PCDD/F soil reservoirs?

Our aim was to identify polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzo-p-furans (PCDFs) in agricultural farmland soils in the Northwest of Mexico. We obtained ≈50 g of soil in five Yaqui Valley (VY) agricultural fields in the north-western Mexican State of Sonora and in five Culiacán Valley (VC) agricultural fields in the north-western Mexican State of Sinaloa. Fields with minimal tillage, with ferti-irrigation, and those with intensive aerial and manual tillage were included. All soil samples were subjected to the chemical activated luciferase gene expression (CALUX(®)) test to determine PCDD/F. On average, samples contained 4.2 ± 1.2 PCDD/F ppt TEQ; VY soil samples contained 4.72 ± 1.23 PCDD/F ppt TEQ, while VC soil samples showed 3.6 ± 1.1 PCDD/F ppt TEQ (p = 0.47). On considering tillage-type, in agricultural fields catalogued as intensive tillage, PCDD/F concentrations were 4.40 ± 0.43 in agricultural fields catalogued as intensive tillage, while in farmlands of another tillage-type these concentrations were slightly higher (5.53 ± 0.8).

[Geometric coding and cluster analysis to assess metabolic control of patients with type 2 diabetes mellitus]. / Codificación geométrica y análisis de conglomerados para evaluar el control metabólico de pacientes con diabetes mellitus tipo 2.

OBJECTIVE: Determine the frequency of combinations of higher-than-normal metabolic control parameters, using geometric coding and hierarchical cluster analysis, in patients with type 2 diabetes (DM2) METHODOLOGY: A descriptive cross-sectional study was conducted in Mexico to assess a group of 1 051 patients with DM2. The inclusion criteria were to have one or more of the following values: fasting glucose of 130 mg/dL, total cholesterol of 240 mg/dL, total triglycerides of 200 mg/dL, Body Mass Index of 27 kg/m(2), and systolic blood pressure higher than 130 mmHg or diastolic blood pressure higher than 85 mmHg. Through geometric coding, the frequencies of all combinations were obtained. Cluster analysis was used to determine similarities among the combinations. RESULTS: Using the proposed instrument, it was observed that the paired combinations with the highest number of subjects were hyperglycemia-hypertriglyceridemia (7.3%) and hyperglycemia-hypercholesterolemia (3.6%). The most frequent polycombinations were hyperglycemia-hypercholesterolemia-hypertriglyceridemia (13.2%) and hyperglycemia-hypertriglyceridemia-hypercholesterolemia-hypertension (10.5%). CONCLUSIONS: Geometric coding and cluster analysis could become a suitable instrument for assessing the metabolic control of patients with DM2, as well as for identifying parameters that will help improve their monitoring and treatment.

[Transcutaneous electrical nervous stimulation in the prognosis of Bell's palsy]. / Estimulación nerviosa eléctrica transcutánea para el pronóstico de parálisis de Bell.

BACKGROUND: The peripheral face palsy (PFP) is the commonest acute cranial neuropathy. The PFP has a showy clinical pattern which contrasts with a favorable course. Our objective was to determine the sensitivity and specificity for the nervous excitability test (NET) with transcutaneous electrical nerve stimulation (TENS) and the time required to obtain face symmetry. METHODS: An analytical cross-sectional study was made in 22 patients with PFP. The goal was the time (days) to obtain face symmetry. The sensitivity and specificity was carried out. RESULTS: A sensitivity and specificity of the NET was of 100 %. The correlation corrected by sex and age between both variables was 0.89. The average in days of recovery was smaller in those with a positive NET (p < 0.05) test. CONCLUSIONS: The test of nervous excitability for PFP with TENS is safe and simple to use in primary care and urgencies services.

[Visceral fat and metabolic manifestations in hypertensive men]. / Grasa visceral y alteraciones metabólicas en hombres hipertensos.

OBJECTIVE: to determine visceral fat and metabolic profile in hypertensive men with and without obesity. METHODS: analytic cross-sectional study was carried out. Thirty hypertensive men with obesity and 30 hypertensive men without obesity from a primary care clinic. Visceral and subcutaneous fat (by computer axial tomography), anthropometry, lipid profile, fibrinogen, and glucose (fasting and 2 hr after glucose load) were measured. RESULTS: hypertensive obese men were younger (48 + or - 7 versus 57 + or - 6 years) who had higher values of blood pressure, lipids and lipoproteins, waist and hip circumference and total abdominal and subcutaneous fat (420 versus 321 cm(2), p < 0.001; and 257 versus 162 cm(2), p = 0.03). Visceral fat was similar in both groups (163 versus 162 cm(2)). The non obese group had higher levels of glucose either fasting and post glucose load. Both groups had increased mean values of fibrinogen and triglycerides and low concentrations of HDL-c. CONCLUSIONS: hypertension was associated with increased visceral fat, regardless the BMI. Although obese hypertensive men were younger, they had a metabolic profile more deleterious than non obese men.

Derivation of a human DOA iPSC line, IISHDOi006-A, with a mutation in the ACO2 gene: c.1999G>A; p.Glu667Lys.

Human iPSC line, IISHDOi006-A, was obtained from fibroblasts of a patient with Dominant Optic Atrophy (DOA) carrying a heterozygous mutation in the gene ACO2: c.1999G>A; p.Glu667Lys. Reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using a non-integrative methodology that involves the use of Sendai virus.

Cardiovascular risk factors and acculturation in Yaquis and Tepehuanos Indians from Mexico.

BACKGROUND: Cardiovascular (CV) risk factors are influenced by behavioral, cultural, and social factors, suggesting that acculturation plays a significant role in the emergency and growth of chronic disease. The objective of this study was to determine the relation between CV risk factors and the main components of acculturation, in Yaquis and Tepehuanos Indians from Mexico. METHODS: This was a cross-sectional population-based study in Yaquis and Tepehuanos communities from the Yaqui Valley in Sonora and the Sierra Madre Occidental Mountains in Durango, in northwest Mexico. Acculturation status is different in both ethnic groups, with Tepehuanos living in small and remote communities retaining their traditional lifestyle and Yaquis living in well-communicated communities that have assumed Westernized lifestyles. RESULTS: A total of 278 indigenous (120 Tepehuanos and 158 Yaquis) were randomly enrolled. Prevalence of obesity (48.1 and 6.7%, p <0.001), diabetes (18.3 and 0.83%, p <0.001), hypertriglyceridemia (43.0 and 15.0%, p <0.001), alcohol consumption (46.8 and 26.6%, p >0.001), and smoking (29.7 and 15.0%, p = 0.006) were significantly higher in Yaquis Indians. High blood pressure (6.3 and 3.3%, p = 0.40) and low HDL-cholesterol (42.4 and 34.2%, p = 0.22) were similar between Yaquis and Tepehuanos. Multivariate regression analysis adjusted by sex and age showed a significant association between calorie intake from saturated fat, but not other nutrients of customary diet, with hyperglycemia (OR 7.4, 95% CI 2.6-20.1), hypertriglyceridemia (OR 3.1, 95% CI 1.5-6.3), and obesity (OR 3.4, 95% CI 1.6-10.1). CONCLUSIONS: Among the components of acculturation, intake of saturated fat is the most strongly associated with the development of CV risk factors.

Toward the Mutational Landscape of Autosomal Dominant Retinitis Pigmentosa: A Comprehensive Analysis of 258 Spanish Families.

Purpose: To provide a comprehensive overview of the molecular basis of autosomal dominant retinitis pigmentosa (adRP) in Spanish families. Thus, we established the molecular characterization rate, gene prevalence, and mutational spectrum in the largest European cohort reported to date. Methods: A total of 258 unrelated Spanish families with a clinical diagnosis of RP and suspected autosomal dominant inheritance were included. Clinical diagnosis was based on complete ophthalmologic examination and family history. Retrospective and prospective analysis of Spanish adRP families was carried out using a combined strategy consisting of classic genetic techniques and next-generation sequencing (NGS) for single-nucleotide variants and copy number variation (CNV) screening. Results: Overall, 60% of our families were genetically solved. Interestingly, 3.1% of the cohort carried pathogenic CNVs. Disease-causing variants were found in an autosomal dominant gene in 55% of the families; however, X-linked and autosomal recessive forms were also identified in 3% and 2%, respectively. Four genes (RHO, PRPF31, RP1, and PRPH2) explained up to 62% of the solved families. Missense changes were most frequently found in adRP-associated genes; however, CNVs represented a relevant disease cause in PRPF31- and CRX-associated forms. Conclusions: Implementation of NGS technologies in the adRP study clearly increased the diagnostic yield compared with classic approaches. Our study outcome expands the spectrum of disease-causing variants, provides accurate data on mutation gene prevalence, and highlights the implication of CNVs as important contributors to adRP etiology.

Chitosan treatment for skin ulcers associated with diabetes.

Infections, ulcerations, gangrene and, in severe cases, extremity amputation, are common complications among diabetic subjects. Various biomaterials have been utilized for the treatment of these lesions. Chitosan is an amino sugar with a low risk of toxicity and immune response. In this study, we evaluated chitosan topical gel and film treatments for subjects with diabetic ulcerations and wounds associated with diabetes mellitus. In a pre-experimental design, we described the result of chitosan gel and film treatment for wounds and skin ulcers among patients with long-standing diabetes mellitus. We studied 8 diabetic patients with wounds and skin ulcers (long duration and Wagner degree 1-2). Initially, most lesions had some degree of infection, tissue damage and ulceration. At the end of the treatment (topical chitosan) period, the infections were cured. All patients experienced a significant improvement in the initial injury and developed granulation tissue and a healthy skin cover. This report represents one of the few published clinical experience regarding the chitosan for the treatment of skin lesions among diabetic subjects. These results are relevant and promising for the treatment of this disease.

Risk factors for three-vessel coronary artery disease in patients of Northwest Mexico.

INTRODUCTION: Three-vessel coronary artery disease is an advanced manifestation of atherosclerosis, with high prevalence in Mexico. OBJECTIVE: The aim of this study was to describe coronary risk factors in a group of patients with three-vessel coronary artery disease in Northwest Mexico. METHODS: A cross sectional study was conducted on a population with three-vessel coronary artery disease from May 2015 to February 2016. The disease was defined when ≥70% stenosis was present in each major epicardial coronary artery. Anthropometric and biochemical parameters were measured in each patient. Ankle-Brachial Index was measured with vascular ultrasound, and Syntax score calculation with an on-line application. Statistical analysis for qualitative differences was performed using Pearson X2 test, with p<0.05 being considered as significant. RESULTS: The study included 100 patients, of whom 75 were male (mean age 63±9 years) and 25 female (mean age 69±9 years). The coronary risk factors observed were diabetes (58%), hypertension (86%), smoking (68%), dyslipidaemia (100%), metabolic syndrome (71%), and obesity/overweight (75%). Diabetes and metabolic syndrome prevalence was higher in women (p=0.03), but smoking was higher in men (76%, p=0.003). Ankle-Brachial Index was abnormal in 58% of patients, the mean Syntax score was in 36.9±11.5, and the prevalence of left main coronary heart disease was 36%. CONCLUSIONS: This group of patients with complex coronary lesions has a high prevalence of coronary risk factors, which could represent a worse prognosis.

Combining targeted panel-based resequencing and copy-number variation analysis for the diagnosis of inherited syndromic retinopathies and associated ciliopathies.

Inherited syndromic retinopathies are a highly heterogeneous group of diseases that involve retinal anomalies and systemic manifestations. They include retinal ciliopathies, other well-defined clinical syndromes presenting with retinal alterations and cases of non-specific multisystemic diseases. The heterogeneity of these conditions makes molecular and clinical characterization of patients challenging in daily clinical practice. We explored the capacity of targeted resequencing and copy-number variation analysis to improve diagnosis of a heterogeneous cohort of 47 patients mainly comprising atypical cases that did not clearly fit a specific clinical diagnosis. Thirty-three likely pathogenic variants were identified in 18 genes (ABCC6, ALMS1, BBS1, BBS2, BBS12, CEP41, CEP290, IFT172, IFT27, MKKS, MYO7A, OTX2, PDZD7, PEX1, RPGRIP1, USH2A, VPS13B, and WDPCP). Molecular findings and additional clinical reassessments made it possible to accurately characterize 14 probands (30% of the total). Notably, clinical refinement of complex phenotypes was achieved in 4 cases, including 2 de novo OTX2-related syndromes, a novel phenotypic association for the ciliary CEP41 gene, and the co-existence of biallelic USH2A variants and a Koolen-de-Vries syndrome-related 17q21.31 microdeletion. We demonstrate that combining next-generation sequencing and CNV analysis is a comprehensive and useful approach to unravel the extensive phenotypic and genotypic complexity of inherited syndromic retinopathies.

[Placental atherosclerosis and markers of endothelial dysfunction in infants born to mothers with gestational diabetes]. / Aterosclerosis placentaria y marcadores de disfunción endotelial en neonatos hijos de madres con diabetes gestacional.

BACKGROUND AND OBJECTIVE: The pathophysiology of gestational diabetes itself causes hyperstimulation of adipose tissue and of the placenta cells increasing the production of inflammatory cytokines, which cause changes in the tissues exposed such as the placenta and foetus. Therefore, the objective of this study was to compare metabolic markers and endothelial dysfunction in umbilical cord blood, as well as to determine the presence of atherosclerosis in the placentas of newborn infants of patients with gestational diabetes and in patients with normally progressing pregnancies. PATIENTS AND METHOD: An analytical cross-sectional study was carried out in 84 patients, obtaining data such as age, smoking and weight gain in pregnancy; the gestational age of the newborns was determined by Capurro, and their weight and destination subsequent to birth, the placentas were also collected in order to look for atherosclerosis through histological studies and glucose, insulin, VLDL-C, HDL-C, triglycerides, cholesterol, fibrinogen, PCR and markers of endothelial dysfunction (adiponectin, VCAM-1, ICAM-1 and IL-6) were determined in blood samples obtained from the umbilical cord. RESULTS: Placental atherosclerosis presented in 28.94% of the group with gestational diabetes compared to 10.52% of the group with normally progressing pregnancies (P=.044); differences were found in glucose, cholesterol, triglycerides, fibrinogen, HOMA-IR, PCR-us, HDL-C, not in VLDL-C. Twenty-one point five percent of the newborns of the gestational diabetes patients required hospitalization, against 5.2% in the control group, CONCLUSIONS: Pregnancies that involve diabetes have higher proportion of atherosclerosis, hospitalization of the newborn, insulin resistance, as well as elevation of markers associated with inflammation and endothelial dysfunction in umbilical cord blood.

Panel-based NGS Reveals Novel Pathogenic Mutations in Autosomal Recessive Retinitis Pigmentosa.

Retinitis pigmentosa (RP) is a group of inherited progressive retinal dystrophies (RD) characterized by photoreceptor degeneration. RP is highly heterogeneous both clinically and genetically, which complicates the identification of causative genes and mutations. Targeted next-generation sequencing (NGS) has been demonstrated to be an effective strategy for the detection of mutations in RP. In our study, an in-house gene panel comprising 75 known RP genes was used to analyze a cohort of 47 unrelated Spanish families pre-classified as autosomal recessive or isolated RP. Disease-causing mutations were found in 27 out of 47 cases achieving a mutation detection rate of 57.4%. In total, 33 pathogenic mutations were identified, 20 of which were novel mutations (60.6%). Furthermore, not only single nucleotide variations but also copy-number variations, including three large deletions in the USH2A and EYS genes, were identified. Finally seven out of 27 families, displaying mutations in the ABCA4, RP1, RP2 and USH2A genes, could be genetically or clinically reclassified. These results demonstrate the potential of our panel-based NGS strategy in RP diagnosis.

A Comprehensive Analysis of Choroideremia: From Genetic Characterization to Clinical Practice.

Choroideremia (CHM) is a rare X-linked disease leading to progressive retinal degeneration resulting in blindness. The disorder is caused by mutations in the CHM gene encoding REP-1 protein, an essential component of the Rab geranylgeranyltransferase (GGTase) complex. In the present study, we evaluated a multi-technique analysis algorithm to describe the mutational spectrum identified in a large cohort of cases and further correlate CHM variants with phenotypic characteristics and biochemical defects of choroideremia patients. Molecular genetic testing led to the characterization of 36 out of 45 unrelated CHM families (80%), allowing the clinical reclassification of four CHM families. Haplotype reconstruction showed independent origins for the recurrent p.Arg293* and p.Lys178Argfs*5 mutations, suggesting the presence of hotspots in CHM, as well as the identification of two different unrelated events involving exon 9 deletion. No certain genotype-phenotype correlation could be established. Furthermore, all the patients´ fibroblasts analyzed presented significantly increased levels of unprenylated Rabs proteins compared to control cells; however, this was not related to the genotype. This research demonstrates the major potential of the algorithm proposed for diagnosis. Our data enhance the importance of establish a differential diagnosis with other retinal dystrophies, supporting the idea of an underestimated prevalence of choroideremia. Moreover, they suggested that the severity of the disorder cannot be exclusively explained by the genotype.

[Helicobacter pylori detection in children with initially negative Gram, Giemsa and Wharting-Starry methods, using other histologic techniques]. / Detección de Helicobacter pylori en niños con los métodos de gram, giemsa y Warthing-Starry, inicialmente negativos con otras técnicas histológicas.

OBJECTIVE: To identify Helicobacter pylori by Gram, Giemsa and Warthing-Starry histological staining methods in biopsies of the digestive tract in children, when Helicobacter pylori identification was negative with hematoxilin and eosin. MATERIAL AND METHODS: We carried out a retrospective, cross-sectional and comparative study of 36 gastric biopsies received in the department of Anatomic-Pathology, in a period often months. All the biopsies were initially stained with routine Hematoxilin and Eosin. Of the 36 biopsies, 24 were negative to Helicobacter pylori. From paraffin blocks of these last biopsies three new histological sections were obtained to dye them with Gram, Giemsa and Warthing-Starry, and two medical pathologists performed the microscopic interpretation of these samples. RESULTS: The search for Helicobacter pylori was carried out in 24 initially negative biopsies, one was positive for Gram (4.16%) and negative for Giemsa and Warthing-Starry and four were both positive for Giemsa and Warthing-Starry (16.66%) and both were also negative for Gram. Only one biopsy was positive for the three methods. The remaining samples were negative for the three stains. CONCLUSION: The Giemsa and/or Warthing-Starry histologic techniques can be a more specific alternative for the determination of Helicobacter pylori in patients with negative digestive tract biopsies with the traditional method of hematoxilin and eosin.