Chiara Di Bella

Angiotensin-Converting Enzyme and Angiotensin Type 2 Receptor Gene Genotype Distributions in Italian Children with Congenital Uropathies

Angiotensin I–converting enzyme (ACE) and angiotensin type 2 receptor (AT2R) gene polymorphisms have been associated with an increased incidence of congenital anomalies of the kidney and urinary tract (CAKUT). We investigated the genotype distribution of these polymorphisms in Italian children with CAKUT. We also evaluated the association between the ACE insertion/deletion and the AT2R gene polymorphisms with the progression of renal damage in subgroups of CAKUT patients. We recruited 102 Italian children with CAKUT; 27 with vesicoureteral reflux; 12 with hypoplastic kidneys; 20 with multicystic dysplastic kidneys; 13 with ureteropelvic junctions stenosis/atresia; 18 with nonobstructed, nonrefluxing primary megaureters; and 12 with posterior urethral valves and compared them with 92 healthy control subjects. ACE and AT2R gene polymorphisms were analyzed by PCR. The identification of AT2R gene polymorphisms in intron 1 and in exon 3 was revealed by enzymatic digestion. ACE genotype distribution in children with CAKUT was no different from that of the control subjects, but the subgroup of patients with radiographic renal parenchymal abnormalities showed an increased occurrence of the D/D genotype. The frequency of the G allele of AT2R gene in children with CAKUT was increased in respect to that of the control subjects. By contrast, no significant difference in the frequency of the C and A alleles of the AT2R gene was found. Our findings indicate that the ACE gene can be a risk factor in the progression of renal parenchymal damage in CAKUT patients. Moreover, a major role of the AT2R gene in the development of CAKUT has been found, at least in Italian children.

A mitochondrial DNA mutation (A3243G mtDNA) in a family with cyclic vomiting

We report the incidence of mitochondrial DNA (mtDNA) mutations and/or deletions in a family from Southern Italy in which four members were affected by a cyclic vomiting syndrome (CVS). The A3243G mtDNA mutation was detected in a boy, his mother, maternal grandmother, and aunt but not in other relatives of the family. Patients with a CVS experience a minimum of three distinct episodes of vomiting and nausea, usually involving more than four emeses in one hour at the peak. They feel quite well between episodes. There is no apparent underlying cause for the vomiting [4]. In the differential diagnosis of CVS, mtDNA mutations should be considered when there is a maternal history of CVS and migraine, clinical findings of seizures, neuromuscular and gastrointestinal symptoms and laboratory evidence of lactate increase [5]. A family was recruited with four members suffering from CVS: a 5-year-old boy, his mother, the maternal grandmother and aunt. The three adults were affected by CVS during childhood, whilst they suffered from migraine at an adult age (Fig. 1). Metabolic investigations revealed permanent hyperlactataemia (5–8 mM) with elevated lactate/pyruvate ratios (L/P) (25–30) in the boy’s mother. In the maternal grandmother and aunt, lactate levels (2.6 and 3.0 mM respectively) were slightly elevated and L/P molar ratios remained in the normal range. The young patient suffered from vomiting fits which lasted for many hours until spontaneous resolution and recurrence with the same characteristics after an interval of 15–20 days. Metabolic acidosis (pH 7.3, bicarbonate 16 mM) with hyperlactataemia (2.5–5.35 mM) and elevated L/P (19–32; normal <16) and ketone body molar ratios were found (2–4.6, normal <2). Hyperalaninaemia, lactic aciduria, and an abnormal excretion of suberic, adipic, and 3-hydroxybutyric acids was observed. Laboratory, clinical and instrumental findings of all four patients excluded organic diseases. To check for point mutations and for a previously described 8.1 kb deletion [3], the region between nucleotides 6687 and 15123 was amplified by the polymerase chain reaction (PCR) in ten overlapping segments and completely sequenced. Screening for the 3243 np mutation in mtDNA was carried out by PCR-RFLP. The relevant region of mtDNA was amplified using the following primers according to the Cambridge sequence [1]: 5’CCCACAGGTCCTAAACTACC-3’ (np 2770–2789) and 5’AGCGAAGGGTTGTAGTAGCC-3’ (np 3456– 3437). The PCR products (10 ll) were then purified and digested with 15 U of the restriction endonuclease ApaI for 1 h at 32 C. The A-to-G mutation at 3243 np created an ApaI restriction site (GAGCCC to GGGCCC) and was easily detected by ethidium bromide staining in a 2.5% agarose gel. In order to detect small amounts of mutated mtDNA in the presence of an excess of wildFig. 1 Pedigree of the family with CVS; the arrow indicates the index case. The percentage of mutated mtDNA in blood and muscle of the patients is I:2=25% and 30%; II:2=35% and 38%; II:3=30% and 32%; III:1=70% and 75% respectively. Affected individuals are indicated by solid symbols Eur J Pediatr (2003) 162: 727–728 DOI 10.1007/s00431-003-1280-1

Identification of α-thalassemia mutations in subjects from Eastern Sicily (Italy) with abnormal hematological indices and normal Hb A2

We analyzed the prevalence of α-thalassemia mutations in 298 subjects from Eastern Sicily (Italy) with reduced mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH), normal HbA2 and HbF, and normal serum iron. In 131 subjects (43.9%) we found six different genotypes of α-thalassemia: -α3.7/αα (36.6%), -α3.7/-α3.7 (27.5%), –MED/αα (10.0%), -α20.5/αα (9.1%), αHphIα/αα (8.4%), αHphIα/αHphIα (6.1%), and -α3.7/αHphIα (2.3%). Our data underline that in Eastern Sicily populations, the molecular screening of α-thalassemia mutations and/or deletions may be useful to better characterize the clinically asymptomatic subjects with a slightly reduced MCV and MCH and normal iron status.

Histological heterogeneity and somatic mtDNA mutations in gastric intraepithelial neoplasia

Somatic mutations of mitochondrial DNA (mtDNA) are associated with various types of human cancer. To elucidate their role in gastric carcinogenesis, we analyzed mutations in the displacement loop region of mtDNA in 24 paraffin-embedded gastric intraepithelial neoplasias (formerly dysplasia) from a high gastric cancer risk area in northern Italy. Helicobacter pylori infection was assessed by histological examination (Giemsa staining). Gastritis was classified according to the guidelines of the Updated Sydney System. The mtDNA displacement loop region was amplified and sequenced from gastric intraepithelial neoplasia samples and adjacent non-neoplastic gastric mucosa. The gastric intraepithelial neoplasias were divided into two groups by their association with H. pylori gastritis. Group A with lesions arising on a background of H. pylori-positive gastritis contained 7 patients, and group B with lesions associated with H. pylori-negative gastritis contained 17 patients. Group A had a larger proportion of high-grade lesions than group B and showed a foveolar phenotype (type II dysplasia). Group B had a larger proportion of cases with mtDNA displacement loop region mutations than group A (P=0.004, Fishers exact test) and exhibited an intestinal phenotype. No evidence of heteroplasmic variants in the mtDNA displacement loop, suggestive of mutations, was detected in gastric biopsies from 25 H. pylori-negative subjects and 60 cancer-unaffected H. pylori-positive patients. These results provide further evidence for the morphologic and mtDNA biomolecular differences of gastric intraepithelial neoplasias, and suggest the existence of two distinct pathways to gastric cancer—corpus-dominant H. pylori gastritis and the atrophy–metaplasia pathway.

Prevalence of Deafness-Associated Connexin-26 (GJB2) and Connexin-30 (GJB6) Pathogenic Alleles in a Large Patient Cohort from Eastern Sicily.

Mutations in the gene encoding the gap junction protein connexin 26 (GJB2) and connexin 30 (GJB6) have been shown to be a major contributor to prelingual, sensorineural, nonsyndromic deafness.

Uteroglobin-related protein 1 gene 112G/A polymorphism and atopic asthma in Sicilian children

The secretory protein, uteroglobin-related protein 1 (UGRP1), is expressed mainly in the lung and trachea and recently has been implicated in asthma. The -112G to A transition in the promoter was reported to be associated with asthma in the Japanese population. However, this has not been replicated in other studies. The aim of this study was to find the association of the UGRP1 gene polymorphism with atopic asthma in the Sicilian population. We conducted a transmission disequilibrium test (TDT) in 73 trios identified through 113 pediatric patients being treated for asthma. A case-control study also was performed by comparing the 113 unrelated asthmatic children and 230 unrelated healthy Italian subjects (121 children and 109 adults). The -112 G/A polymorphism was genotyped by the polymerase chain reaction-restriction fragment length polymorphism method and direct sequencing. The TDT revealed that the -112A allele was not preferentially transmitted from the parents to asthmatic offspring (chi-square = 3.08; p = NS). Neither the presence of at least one A allele in an individuals genotype (sum of the G/A and A/A genotype) nor the -112A allele was more prevalent among the asthma subjects than among the control subjects. Our results suggest that the -112G/A polymorphism does not play a significant role in the genetic predisposition of the UGRP1 gene in atopic asthma in the Sicilian population.

Genetic and clinical aspects of Wolfram syndrome 1, a severe neurodegenerative disease

Wolfram syndrome 1 (WS1) is a rare autosomal recessive neurodegenerative disease characterized by diabetes insipidus, diabetes mellitus, optic atrophy, deafness, and other abnormalities. WS1 usually results in death before the age of 50 years. The pathogenesis of WS1 is ascribed to mutations of human WFS1 gene on chromosome 4p encoding a transmembrane protein called wolframin which has physiological functions in membrane trafficking, secretion, processing, and/or regulation of ER calcium homeostasis. Different types of WFS1 mutations have been identified, and some of these have been associated with a dominant, severe type of WS.

Correction: Genetic and clinical aspects of Wolfram syndrome 1, a severe neurodegenerative disease

The original version of this Article erroneously cropped part of the abstract. The abstract has now been corrected to read ‘Wolfram syndrome 1 (WS1) is a rare autosomal recessive neurodegenerative disease characterized by diabetes insipidus, diabetes mellitus, optic atrophy, deafness, and other abnormalities. WS1 usually results in death before the age of 50 years. The pathogenesis of WS1 is ascribed to mutations of human WFS1 gene on chromosome 4p encoding a transmembrane protein called wolframin, which has physiological functions in membrane trafficking, secretion, processing, and/or regulation of ER calcium homeostasis. Different types of WFS1 mutations have been identified, and some of these have been associated with a dominant, severe type of WS. Mutations of CISD2 gene cause autosomal recessive Wolfram syndrome 2 (WS2) characterized by the absence of diabetes insipidus and psychiatric disorders, and by bleeding upper intestinal ulcer and defective platelet aggregation. Other WFS1-related disorders such as DFNA6/14/38 nonsyndromic low-frequency sensorineural hearing loss and Wolfram syndrome-like disease with autosomal dominant transmission have been described. WS1 is a devastating disease for the patients and their families. Thus, early diagnosis is imperative to enable proper prognostication, prevent complications, and reduce the transmission to further progeny. Although there is currently no effective therapy, potential new drugs have been introduced, attempting to improve the progression of this fatal disease.” in both the PDF and HTML versions of the Article.

A Novel Mutation of the δ-Globin Gene in an Asymptomatic 30-Year-Old Female

hinder the diagnosis of the β-thalassemia trait [4]. Values of HbA2 >3.2% associated with microcytosis characterize the β-thalassemia trait [3]. However, the coinheritance of β-globin and δ-globin gene mutations can decrease HbA2 values to normal or lower levels due to a decreased δ-chain production. As iron deficiency anemia is also characterized by low HbA2 levels, this disease must firstly be ruled out by evaluating iron parameters. Subsequently, a δ-globin mutation screening should be performed, especially in subjects from geographic areas with a high incidence of β-thalassemia carriers. δ-Thalassemia (OMIM No. 142000) resulting from mutations on the HBD gene usually has no clinical consequences, but it may cause a reduced rate of hemoglobin A2 (HbA2) [1, 2]. Human HbA2 (α2δ2), which contains δ-globin, represents a minor fraction of the Hb found in human adults. In normal individuals, it constitutes between 2.5 and 3.3% of the total adult Hb content as measured by high-performance liquid chromatography [3]. Although genetic defects (δ-thalassemia o HbA2 variants) that decrease HbA2 levels (2% in heterozygote or ≤0.6% in homozygote subjects) do not affect health, they can Received: June 22, 2017 Accepted after revision: November 6, 2017 Published online: January 13, 2018

Co-inheritance of Hb Hershey [β70(E14) Ala → Gly] and Hb La Pommeraie [β133(H11)Val → Met] in a Sicilian subject

Objectives: This report represents the first observation in Sicily of two rare β‐globin gene variants, Hb Hershey [β70(E14) Ala→Gly] and Hb La Pommeraie [β133(H11)Val→Met], found in a 35‐year‐old male patient from Messina, in the north‐east of Sicily during population screening for hemoglobinopathies. Methods: The occurrence of the Hb variants was assessed by cation exchange chromatography while complete blood counts were obtained using automatic cell counters. Red cell lysates were analyzed by electrophoresis at alkaline and acid pH. Stability of hemoglobin was checked by the isopropanol precipitation test and by the heat tests while inclusion bodies and reticulocyte count were determined by incubation of blood samples with brilliant cresyl blue. Molecular analysis was performed by DNA sequencing of β‐ and α‐globin genes. Results: We observed an abnormally high performance liquid chromatography elution with a slight reduction in mean corpuscular volume and mean corpuscular haemoglobin parameters and mutations at codon 70 GCC→GGC (Hb Hershey) and at codon 133 GTG→ATG (Hb La Pommeraie) in β‐globin gene. Conclusion: Family analysis of three generations demonstrated the presence of these two mutations in trans. So it was possible to describe the phenotypes of these variants in a heterozygous state and in double heterozygous state.