Identification of a molecular defect in a stillborn fetus with perinatal lethal hypophosphatasia using a disease-associated genome sequencing approach.

Lethal skeletal disorders represent a heterogeneous and clinically variable group of genetic conditions, usually difficult to diagnose without post-mortem radiological assessment. Here we report on a stillborn patient delivered at 22 weeks of gestation who presented with severe skeletal symptoms comprising limb shortening and intrauterine fractures detected upon prenatal ultrasound and autopsy examination. Since post-mortem X-ray was refused and no phenotypic diagnosis could be attempted, we performed next-generation sequencing (NGS) of 2741 genes associated with all known Mendelian disorders. With this strategy, we were able to demonstrate the diagnosis at a molecular level, which turned out to be perinatal lethal hypophosphatasia (HPP). This severe form of HPP represents an inborn defect of ossification often resulting in stillbirth or postnatal death. The NGS panel revealed compound heterozygous ALPL missense mutations: c.1283G>C(p.Arg428Pro) and c.1363G>A(p.Gly455Ser). Mutations detected in our case, although previously described in other patients, have not been reported to co-occur in a single individual. The diagnosis established in our index using the NGS-based approach could have been successfully reached by standard radiography. Thus, our report points to the importance of X-ray examination in stillborn cases and highlights the emerging role of NGS strategies in the diagnostic process of prenatally manifesting skeletal disorders.

Results from the Survey of Antibiotic Resistance (SOAR) 2011-13 in the Gulf States.

OBJECTIVES: To provide surveillance data on the susceptibility of community-acquired respiratory tract isolates from four Gulf and Near East countries from 2011 to 2013. METHODS: MICs were determined using Etests(®) for all antibiotics evaluated except erythromycin, where testing was by disc diffusion. Susceptibility was assessed using CLSI, EUCAST and pharmacokinetic/pharmacodynamic (PK/PD) breakpoints. RESULTS: Seven hundred and twenty-six respiratory isolates comprising 265 isolates of Streptococcus pneumoniae, 125 isolates of Streptococcus pyogenes and 336 isolates of Haemophilus influenzae were collected from Bahrain, Lebanon, Oman and the United Arab Emirates (UAE). Among S. pneumoniae, susceptibility to penicillin was low in the UAE and Bahrain. Macrolide susceptibility was ∼45%-60% in the UAE and Oman but higher in Lebanon (73.7%) and Bahrain (84%-85%). Penicillin susceptibility using CLSI intravenous breakpoints was >85% in all countries. Antibiotic susceptibility of S. pneumoniae was lower in UAE and Oman. Among S. pyogenes isolates, resistance to erythromycin was highest in Oman (31.6%) but <20% in the other countries. In H. influenzae, susceptibility to most antibiotics was high, except for ampicillin in Lebanon (70.2%) and amoxicillin in Oman (95.4%). Lebanon also had a high percentage (14.9%) of ß-lactamase-positive isolates with non-susceptibility to ampicillin. Amoxicillin/clavulanic acid susceptibility was >95% in all countries. Use of EUCAST versus CLSI breakpoints demonstrated profound differences for cefaclor and cefuroxime in S. pneumoniae and H. influenzae, with EUCAST showing lower susceptibility. CONCLUSIONS: There was considerable variability in susceptibility among countries in the same region. Thus, continued surveillance is necessary to track future changes in antibiotic resistance.

A novel nonsense mutation in CUL4B gene in three brothers with X-linked mental retardation syndrome.

Cabezas syndrome (MIM 300354) is a recently identified syndromic form of X-linked mental retardation (XLMR) caused by mutations in the CUL4B gene. In total, nine XLMR families carrying mutations in the CUL4B gene have been described to date. Here, we present a detailed clinical phenotype of three affected brothers of Polish descent. Based on the symptoms, we made a clinical diagnosis of Cabezas syndrome, which was subsequently confirmed by identification of a novel nonsense mutation (c.2107A-->T, p.703K-->X) in exon 18 of the CUL4B gene. The mutation was inherited from an asymptomatic mother and was present in all three affected brothers. The patients presented with typical features of Cabezas syndrome, such as severe mental retardation, speech impairment, hyperactivity, seizures, intention tremor, inguinal hernia, small feet, and craniofacial dysmorphism. In addition to previously described symptoms, syndactyly of the second and third toes and skin manifestations (hyperhydrosis and keratosis pilaris) were present in our cases. Our report provides further support that Cabezas syndrome is a recognizable syndromic form of XLMR. We conclude that the CUL4B gene should be screened in males with severe speech impairment and primary intention tremor, especially if characteristic facial dysmorphism is also present.

A new sporadic case of early-onset Loeys-Dietz syndrome due to the recurrent mutation p.R528C in the TGFBR2 gene substantiates interindividual clinical variability.

We report on a 2-year-old Polish girl with typical manifestations of Loeys-Dietz syndrome (LDS), a rare genetic condition belonging to the group of Marfan-related disorders. The characteristic LDS symptoms observed in the girl included craniofacial dysmorphism (craniosynostosis, cleft palate, hypertelorism), arachnodactyly, camptodactyly, scoliosis, joint laxity, talipes equinovarus, translucent and hyperelastic skin, and umbilical hernia. Mild dilatation of the ascending aorta and tortuous course of the left internal carotid artery were recognized during her second year of life. Molecular genetic testing revealed a heterozygous missense mutation (c.1582C>T, p.R528C) in the transforming growth factor beta receptor II gene (TGFBR2). This mutation has been previously associated with LDS in 5 unrelated cases, and was never reported in patients with other Marfan-related disorders. Comparison of the phenotypes of our patient and these 5 individuals with c.1582C>T showed that only the hallmark triad of the syndrome - consisting of hypertelorism, aortic root dilatation/aneurysm, and cleft palate or bifid uvula - was present in all 6 cases. Interestingly, none of the 5 individuals who underwent psychological evaluation showed developmental delay. The pattern of all other LDS features showed interindividual variability. Our data support the recently reported observation that symptoms of LDS can develop at a very young age, making early diagnosis and management essential for these patients. This is the first report on a Polish infant with typical LDS symptoms caused by a TGFBR2 mutation.

A novel GJA1 missense mutation in a Polish child with oculodentodigital dysplasia.

Oculodentodigital dysplasia (ODDD) (OMIM #164200) is a rare congenital, autosomal dominant disorder comprising craniofacial, ocular, dental, and digital anomalies. The syndrome is caused by GJA1 mutations. The clinical phenotype of ODDD involves a characteristic dysmorphic facies, ocular findings (microphthalmia, microcornea, glaucoma), syndactyly type III of the hands, phalangeal abnormalities, diffuse skeletal dysplasia, enamel dysplasia, and hypotrichosis. In a Polish child with the clinical symptoms typical of ODDD, we demonstrated a novel missense mutation c.C31A resulting in p.L11F substitution. Our report provides evidence on the importance of this highly conserved amino acid residue for the proper functioning of GJA1 protein.

Molecular typing of methicillin-resistant Staphylococcus aureus isolated in a Bahrain hospital.

OBJECTIVE: To investigate antibacterial resistance patterns and genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) obtained at the Salmaniya Medical Complex in Bahrain. METHODS: A total of 53 consecutive MRSA isolates obtained from 53 patients were studied using antibacterial resistance patterns, coagulase gene polymorphism, staphylococcal cassette chromosome mec (SCCmec) typing and pulsed-field gel electrophoresis (PFGE). RESULTS: There was a high prevalence of resistance to fusidic acid (92.5%), ciprofloxacin (92.5%), erythromycin (90.6%), tetracycline (88.7%), trimethoprim (88.7%), streptomycin (88.7%), kanamycin (83.0%) and gentamicin (73.6%). Coagulase gene typing divided the isolates into five coagulase types comprising coagulase type 36 (86.7%), type 20 (3.8%), type 16 (3.8%), type 38 (1.9%) and type 384 (3.8%). They belonged to SCCmec type III (86.7%) and SCCmec type IV (13.3%). PFGE identified five pulsotypes (types A-E) with PFGE type A and its subtypes comprising 83% of the isolates. PFGE type A isolates were multiresistant and had the SCCmec type III and coagulase type 36 genotype. The SCCmec type IV isolates were nonmultiresistant with different genetic backgrounds. CONCLUSIONS: The study identified two types of MRSA in the hospital during the study period. One type consisted of a persistent multiresistant PFGE clone with the SCCmec type III and coagulase type 36 genotypes. The second type consisted of nonmultiresistant isolates that belonged to different genetic backgrounds and were isolated less frequently.

Impact of introducing quality control/quality assurance (QC/QA) guidelines in respiratory specimen processing.

OBJECTIVES: To assess the impact of the introduction of a new quality control/quality assurance (QA/QC) protocol on the processing and reporting of respiratory specimens. METHODS: After implementation of guidelines for processing respiratory specimens, an investigation was carried out over a six-month period on 200 specimens, 105 sputa, and 95 deep tracheal aspirates (DTAs), assessed blindly by two independent investigators. Data regarding disagreement were arranged into two subgroups. A minor disagreement was defined as a difference in the two assessments of < 10 or 10-25 for white blood cell (WBC) or squamous epithelial cell (SEC) counts. A major disagreement was defined as one assessor reporting < 10 and the other > 25 for either WBC count or SEC count, or one assessor reporting the specimen as non-assessable, or both assessors having a minor disagreement in both the WBC count and the SEC count. RESULTS: Agreement was obtained on 111 samples. For 45 specimens, a major disagreement was documented, and in 44 cases, a minor disagreement was recorded, WBC being the most common cause of divergence. Data for sputa and DTAs were examined separately: of 45 major disagreements, 64.4% were observed for DTAs, while minor disagreements were recorded mostly for sputa. The role of the settings in which samples were taken in affecting quality was studied. Among the 105 sputa, 27 were from Community Health Centers (CHCs) and 78 from hospitalized patients. Agreement between the two observers was obtained in 48.1% of CHC cases versus 60.2% of hospital samples. To investigate how many of the rejected samples presented WBC > 25 suggestive of infection, we looked at the 107 samples rejected during the six-month period, grouped according to the suspected diagnosis. The highest number of rejected samples falls in the category of unrelated (non-respiratory) diagnosis, and clinical suspicion is not helpful in Gram stain interpretation. The annual saving (not culturing, not testing, and not treating) derived from this simple QC procedure totals about 5000 Euro. CONCLUSIONS: Standardization in microscopic screening of respiratory samples is difficult to achieve. Criteria for rejection must be adapted to local conditions after discussion with clinicians to increase their compliance with the newly introduced guidelines and to avoid sending unnecessary specimens. The effects on patient management and cost control are significant.