Mutations in Radial Spoke Head Genes and Ultrastructural Cilia Defects in East-European Cohort of Primary Ciliary Dyskinesia Patients

Primary ciliary dyskinesia (PCD) is a rare (1/20,000), multisystem disease with a complex phenotype caused by the impaired motility of cilia/flagella, usually related to ultrastructural defects of these organelles. Mutations in genes encoding radial spoke head (RSPH) proteins, elements of the ciliary ultrastructure, have been recently described. However, the relative involvement of RSPH genes in PCD pathogenesis remained unknown, due to a small number of PCD families examined for mutations in these genes. The purpose of this study was to estimate the involvement of RSPH4A and RSPH9 in PCD pathogenesis among East Europeans (West Slavs), and to shed more light on ultrastructural ciliary defects caused by mutations in these genes. The coding sequences of RSPH4A and RSPH9 were screened in PCD patients from 184 families, using single strand conformational polymorphism analysis and sequencing. Two previously described (Q109X; R490X) and two new RSPH4A mutations (W356X; IVS3_2–5del), in/around exons 1 and 3, were identified; no mutations were found in RSPH9. We estimate that mutations in RSPH4A, but not in RSPH9, are responsible for 2–3% of cases in the East European PCD population (4% in PCD families without situs inversus; 11% in families preselected for microtubular defects). Analysis of the SNP-haplotype background provided insight into the ancestry of repetitively found mutations (Q109X; R490X; IVS3_2–5del), but further studies involving other PCD cohorts are required to elucidate whether these mutations are specific for Slavic people or spread among other European populations. Ultrastructural defects associated with the mutations were analyzed in the transmission electron microscope images; almost half of the ciliary cross-sections examined in patients with RSPH4A mutations had the microtubule transposition phenotype (9+0 and 8+1 pattern). While microtubule transposition was a prevalent ultrastructural defect in cilia from patients with RSPH4A mutations, similar defects were also observed in PCD patients with mutations in other genes

Towards a human rights-based contraceptive policy, a critique of anti-sterilisation law in Poland

grantor: University of TorontoSterilisation is one of the safest, most effective and most widely used method of family planning in the world. However, it is illegal and inaccessible in Poland. This thesis argues that there are certain harms imposed by the anti-sterilisation policy in Poland, and that they amount to a violation of human rights. It argues that limiting access to a comprehensive range of contraceptive option is harmful for reproductive and sexual well-being. Certain concerns associated with contraceptive sterilisation (e.g. fear of abuse or post-sterilisation regret) would be addressed more appropriately by less restrictive measures that respect rights of individuals and better respond to their needs. I argue that an array of contraceptive options that is as comprehensive as medical technology and modern health policy will allow is a necessary component of equality for women. As such, sterilisation policy should be part of the comprehensive reproductive health policy, which should be built upon principles of equality and public health rather than on moralities, myths and popular misconceptions about the people's capacity to control fertility.L.L.M

CFTR mutations spectrum and the efficiency of molecular diagnostics in Polish cystic fibrosis patients.

Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane regulator gene (CFTR). In light of the strong allelic heterogeneity and regional specificity of the mutation spectrum, the strategy of molecular diagnostics and counseling in CF requires genetic tests to reflect the frequency profile characteristic for a given population. The goal of the study was to provide an updated comprehensive estimation of the distribution of CFTR mutations in Polish CF patients and to assess the effectiveness of INNOLiPA_CFTR tests in Polish population. The analyzed cohort consisted of 738 patients with the clinically confirmed CF diagnosis, prescreened for molecular defects using INNOLiPA_CFTR panels from Innogenetics. A combined efficiency of INNOLiPA CFTR_19 and CFTR_17_TnUpdate tests was 75.5%; both mutations were detected in 68.2%, and one mutation in 14.8% of the affected individuals. The group composed of all the patients with only one or with no mutation detected (109 and 126 individuals, respectively) was analyzed further using a mutation screening approach, i.e. SSCP/HD (single strand conformational polymorphism/heteroduplex) analysis of PCR products followed by sequencing of the coding sequence. As a result, 53 more mutations were found in 97 patients. The overall efficiency of the CF allele detection was 82.5% (7.0% increase compared to INNOLiPA tests alone). The distribution of the most frequent mutations in Poland was assessed. Most of the mutations repetitively found in Polish patients had been previously described in other European populations. The most frequent mutated allele, F508del, represented 54.5% of Polish CF chromosomes. Another eight mutations had frequencies over 1%, 24 had frequencies between 1 and 0.1%; c.2052-2053insA and c.3468+2_3468+3insT were the most frequent non-INNOLiPA mutations. Mutation distribution described herein is also relevant to the Polish diaspora. Our study also demonstrates that the reported efficiency of mutation detection strongly depends on the diagnostic experience of referring health centers

Cystic fibrosis diagnosed in 36-year old man

Cystic fibrosis(CF)is the most common autosomal recessive disorder in Caucasians. There is considerable variability in the clinical presentation and course of cystic fibrosis. Adult patients with mild symptoms are increasingly being diagnosed with CF. We present a case of a man diagnosed with CF at age 36 years. The diagnosis was suspected because of recurrent nasal polyposis, bronchiectases, male infertility, and a positive sweat tests. One CFTR mutation was identified. CONCLUSIONS: 1. Cystic fibrosis should be included into the differential diagnosis of chronic respiratory symptoms in adults. 2. CF patients diagnosed in adulthood usually have milder clinical course of the disease

Mutation detection efficiency using INNOLiPA tests<i>: CFTR19</i> and <i>CFTR17TnUpdate</i>.

<p>Legend: ^a I148T included in CFTR-17TnUpdate was not counted as a mutation if not in cis with c.3067-72del6 (l.n. 3199del6); c.1210(-12)T_n site in intron 9 (l.n.IVS8-T_n) without data on the associated TG repeat was not counted as a mutation.</p

Mutations found in the analyzed cohort of 738 Polish CF patients, sorted according to the position in the gene.

<p>Legend: ^a IL19 i 17 – mutations included in the INNOLiPA tests (see below); CFMDB – non-INNOLiPA mutations present in the CTFR mutation database; novel – mutations first reported in this study; ^b in three chromosomes R668C with G576A in trans; ^c F508del, c.1585-1G>A, G542X, N1303K or c.579+3A>G; ^d F508del, G542X, R553X or N1303K; ^e not pathogenic if not in cis with c.3067-72del6 (l.n.3199del6); ^f not pathogenic – see explanation the text; ^g not pathogenic if not in cis with G1244V.</p>a<p>Mutations detected by two INNOLiPA_CFTR tests (legacy names): IL19 (INNOLiPA_<i>CFTR</i>19): F508del; G542X; N1303K; W1282X; G551D; 1717-1G>A; R553X; <i>CFTR</i>dele2,3(21kb); I507del; 711+1G>T; 3272-26A>G; 3905insT; R560T; 1898+1G>A; S1251N; I148T; 3199del6; 3120+1G>A; Q552X.</p><p>IL17 (INNOLiPA_<i>CFTR</i>17_TnUpdate): 621+1G>T; 3849+10kbC>T; 2183AA>G; 394delTT; 2789+5G>A; R1162X; 3659delC; R117H; R334W; R347P; G85E; 1078delT; A455E; 2143delT; E60X; 2184delA; 711+5G>A; polymorphism 5T/7T/9T.</p

Comparison of the efficiency of mutation detection efficiency using INNOLiPA tests and teh follow-up screening (SSCP/HD and sequencing).

<p>Legend: ^a including six alleles with T₅_TG_12–13 in intron 9; ^b including four alleles with T_5or8_TG_12–13 in intron 9; ^c also analyzed by MLPA</p

Distribution of the most frequent CFTR mutations detected in CF patients from Poland, compared with Central and Southeastern European populations.

<p>Legend: Data are given in %. ^a non-INNOLiPA mutations are in bold, and a novel mutation is underlined; ^b very close to the earlier estimate of 54% based on a much smaller study group of PL patients <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Aznarez1" target="_blank">[7]</a>; ^c only selected segments of the gene have been screened; ^d frequency significantly higher or ^e lower than in Polish cohort (p<0.005, Pearsons’s chi square); ^f<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Bobadilla1" target="_blank">[5]</a>; NA– not analyzed/not available; Poland (mostly Southern and Western Poland; this study); Czech Republic <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Krenkova1" target="_blank">[21]</a>; Slovakia (based on <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Krenkova1" target="_blank">[21]</a>); Germany <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Dork1" target="_blank">[22]</a>; Lithuania <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Giannattasio1" target="_blank">[23]</a>; Western Ukraine <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Makukh1" target="_blank">[24]</a>; East Hungary <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Ivady1" target="_blank">[25]</a>; Romania <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Frentescu1" target="_blank">[26]</a>; Bulgaria <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Angelicheva1" target="_blank">[27]</a>; Serbia <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Radivojevic1" target="_blank">[28]</a>; Greece <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089094#pone.0089094-Kanavakis1" target="_blank">[29]</a>.</p

<i>CFTR</i> mutation detection efficiency in PL CF patients from different health centers.

<p>A. Both mutations identified. B. One mutation identified. C. No mutation identified. Diagonal stripes – INNOLiPA mutations; solid black – non-INNOLiPA mutations. Rabka – Institute of Tuberculosis and Lung Diseases in Rabka; Other – other health care centers in Poland.</p

Analysis of exposure to Hg from the perspective of interaction between elements

Background. The aim of the study has been to determine mercury content in selected biological samples: pharyngeal tonsils, gallbladder and femoral head tissues. Methods. Hg content in pharyngeal tonsils (n474), deposits of the gallbladder (n180) in femoral head tissues (n319) has been determined by ICP-AES method with the use of the plasma spectrophotometer Optima 5300DV. Biological samples were dissolved by nitric acid (V) in pressure digestion system in a closed system PD 3-6. Results. Co-occurrence of Hg with other elements was assessed based on two way correlation analysis results. The analysis of mercury co-occurrence with other elements in exemplary biological samples was carried out for samples in which Hg content generally ranged: in femur bone tissue from 0,02–0,97 μg/g; in hydroxyapatites of deposits of the gallbladder from 0,02–0,99 μg/g and in pharyngeal tonsils of children from 0,05 to 5,85 μg/g. Because pharyngeal tonsils are located in the main stream of inhaled air where the highest concentration occurred, the highest accumulation of mercury Hg with other trace elements also occurred. The number of significant accumulations is the greatest and the results are very important in the risk evaluation of two way correlation analysis concerning femoral head tissues. In deposits of the gallbladder the changes of mercury content significantly correlated directly proportional (r0,80 p0,005) Ag, Sr, Ba, Se, Cu, Al in women and in men Cu (r0,80 p0,001). Conclusion. Biological availability of some elements such as As, Se, Be, Sb had a big impact on Hg content in single investigated tissues. The choice of a given biological sample, for risk assessment of Hg, ought to take into account pilot studies results on how mercury co-occurs with other metals