No abstract

Congenital hemolytic anemias (CHAs) are heterogeneous and rare disorders caused by alterations in structure, membrane transport, metabolism, or red blood cell production. The pathophysiology of these diseases, in particular the rarest, is often poorly understood, and easy-to-apply tools for diagnosis, clinical management, and patient stratification are still lacking. We report the 3-years monocentric experience with a 43 genes targeted Next Generation Sequencing (t-NGS) panel in diagnosis of CHAs; 122 patients from 105 unrelated families were investigated and the results compared with conventional laboratory pathway. Patients were divided in two groups: 1) cases diagnosed with hematologic investigations to be confirmed at molecular level, and 2) patients with unexplained anemia after extensive hematologic investigation. The overall sensitivity of t-NGS was 74 and 35% for families of groups 1 and 2, respectively. Inside this cohort of patients we identified 26 new pathogenic variants confirmed by functional evidence. The implementation of laboratory work-up with t-NGS increased the number of diagnoses in cases with unexplained anemia; cytoskeleton defects are well detected by conventional tools, deserving t-NGS to atypical cases; the diagnosis of Gardos channelopathy, some enzyme deficiencies, familial siterosterolemia, X-linked defects in females and other rare and ultra-rare diseases definitely benefits of t-NGS approaches.

Maternal cbl status is an important determinant of infantile cbl status. Both maternal holoTC and tHcy may assist in predicting infantile cbl status. The finding of high prevalence of maternal and infantile cbl deficiency in this study points towards the need for effective strategies to prevent cbl deficiency in women prior to getting pregnant.

Few studies have examined the association between maternal vitamin B 12 status and their breast-fed infants' findings. The objective of this study was to analyze the association of maternal B 12 status with infant findings including neurodevelopmental outcome in breast-fed babies with B 12 deficiency. Correlation analyses between the laboratory findings of infants with B 12 deficiency (n = 120) and their mothers were performed and the association of maternal B 12 status with infant findings including the Denver-II developmental screening test (DDST II) results was examined. There was a significant correlation between infant and maternal B 12 levels (r = 0.222; P = 0.030), and between infant and maternal homocysteine (Hcy) levels (r = 0.390; P < 0.001). Among the babies 4 months of age or older, maternal Hcy levels were significantly correlated with infant mean corpuscular hemoglobin (r = 0.404; P = 0.001) and infant mean corpuscular volume (r = 0.461; P < 0.001). Mothers of infants with abnormal DDST II had lower vitamin B 12 (196.9 ± 41.2 vs. 247.0 ± 77.0 pg/mL; P = 0.018) and higher Hcy levels (17.3 ± 5.0 vs. 10.7 ± 3.1 µmol/L; P < 0.001) than mothers of infants with normal DDST II. A lower maternal vitamin B 12 status may be related to impaired neurodevelopment in breast-fed infants with vitamin B 12 deficiency. Pregnant and lactating women should be offered easy access to healthy nutrition and vitamin B 12 supplements.

The diagnosis of mild bleeding disorders is not easy as most of the "healthy" individuals also report bleeding symptoms. In order to get a precise bleeding history, Pediatric Bleeding Questionnaire (PBQ) has been developed. In our study, Turkish children diagnosed with Von Willebrand disease (VWD), platelet function defect (PFD), and healthy children without any symptoms (control group 1) and healthy children with symptoms but found hemostatically normal (control group 2) were analyzed with PBQ. The cut off level for "positive bleeding score" was found to be ≥2 (area under the curve [AUC]: 0.785, 95% confidence interval [CI]: 0.718-0.852). The sensitivity, specificity, positive predictive value, and negative predictive value of PBQ to define VWD versus control group 1 was 100%, 97.4%, 96.4%, and 100%; VWD versus control group 2 was 100%, 53.1%, 64.3%, and 100%; PFD versus control group 1 was 93.3%, 53.1%, 73.7%, and 85%; and PFD versus control group 2 was 93.3%, 53.1%, 73.7%, and 85%, respectively.