Current Status of Newborn Screening in Southeastern Europe.

Significant part of Southeastern Europe (with a population of 76 million) has newborn screening (NBS) programs non-harmonized with developed European countries. Initial survey was conducted in 2013/2014 among 11 countries from the region (Albania, Bulgaria, Bosnia and Herzegovina (BIH), Croatia, Kosovo, Macedonia, Moldova, Montenegro, Romania, Serbia, and Slovenia) to assess the main characteristics of their NBS programs and their future plans. Their cumulative population at that time was ~52,5 million. At that time, none of the countries had an expanded NBS program, while phenylketonuria screening was not introduced in four and congenital hypothyroidism in three of 11 countries. We repeated the survey in 2020 inviting the same 11 countries, adding Cyprus, Greece, Hungary, and Malta (due to their geographical position in the wider region). The aims were to assess the current state, to evaluate the change in the period, and to identify the main obstacles impacting the implementation of expanded NBS and/or reaching a wider population. Responses were collected from 12 countries (BIH-Federation of BIH, BIH-Republic of Srpska, Bulgaria, Croatia, Greece, Hungary, Kosovo, North Macedonia, Malta, Montenegro, Romania, Serbia, Slovenia) with a population of 68.5 million. The results of the survey showed that the regional situation regarding NBS only modestly improved in this period. All of the surveyed countries except Kosovo screened for at least congenital hypothyroidism, while phenylketonuria was not screened in four of 12 countries. Croatia and Slovenia implemented an expanded NBS program using tandem mass spectrometry from the time of last survey. In conclusion, the current status of NBS programs in Southeastern Europe is very variable and is still underdeveloped (or even non-existent) in some of the countries. We suggest establishing an international task-force to assist with implementation and harmonization of basic NBS services where needed.

Neonatal Screening in Europe Revisited: An ISNS Perspective on the Current State and Developments Since 2010.

Neonatal screening (NBS) was initiated in Europe during the 1960s with the screening for phenylketonuria. The panel of screened disorders ("conditions") then gradually expanded, with a boost in the late 1990s with the introduction of tandem mass spectrometry (MS/MS), making it possible to screen for 40-50 conditions using a single blood spot. The most recent additions to screening programmes (screening for cystic fibrosis, severe combined immunodeficiency and spinal muscular atrophy) were assisted by or realised through the introduction of molecular technologies. For this survey, we collected data from 51 European countries. We report the developments between 2010 and 2020 and highlight the achievements reached with the progress made in this period. We also identify areas where further progress can be made, mainly by exchanging knowledge and learning from experiences in neighbouring countries. Between 2010 and 2020, most NBS programmes in geographical Europe matured considerably, both in terms of methodology (modernised) and with regard to the panel of conditions screened (expanded). These developments indicate that more collaboration in Europe through European organisations is gaining momentum. We can only accomplish the timely detection of newborn infants potentially suffering from one of the many rare diseases and take appropriate action by working together.

Role of omega-3 polyunsaturated fatty acids in gestational diabetes, maternal and fetal insights: current use and future directions.

ω-3-Polyunsaturated fatty acids (ω-3 PUFAs) are widely used during pregnancy and gestational diabetes mellitus (GDM). ω-3 PUFAs are beneficial in the regulation of maternal and fetal metabolic function, inflammation, immunity, macrosomia (MAC), oxidative stress, preeclampsia, intrauterine growth, preterm birth, offspring metabolic function, and neurodevelopment. Dietary counseling is vital for improving therapeutic outcomes in patients with GDM. In maternal circulation, ω-3 PUFAs are transported via transporters, synthesis enzymes, and intracellular proteins, which activate nuclear receptors and play central roles in the cellular metabolic processes of placental trophoblasts. In patients with GDM, this process is compromised due to abnormal functioning of the placenta, which disrupts the normal mother to fetus transport. This results in reduced fetal levels of ω-3 PUFAs, which contributes negatively to fetal growth, metabolic function, and development. Dietary counseling and nutritional assessment remain challenging in the prevention and alleviation of GDM. Therefore, personalized approaches, including measurement of the ω-3 index, pharmacogenetic implementation strategies, and appropriate supplementation with ω-3 PUFAs are used to achieve sufficient distribution in the maternal and fetal fluids during the entire pregnancy period. Developing new dosing guidelines and personalized approaches, determining the mechanisms of ω-3 PUFAs in the placenta, and examining the pharmacodynamic and pharmacokinetics interactions involving ω-3 PUFAs will lead to better management and increase the quality of life of patients with GDM and their offspring. Moreover, different strategies for supplementing with ω-3 PUFAs, improving their placental transport, and pharmacological exploration of the maternal-fetal interactions will help to further elucidate the role of ω-3 PUFAs in women with GDM. In this review, we summarize the current information on the potential therapeutic benefits and clinical applicability of ω-3 PUFAs in patients with GDM and their offspring, highlighting recent progress and future perspectives in this field. Studies investigating the mechanisms of ω-3 PUFA transport to targeted tissues have spurred an interest in personalized treatment strategies for patients with GDM and their offspring. To implement such therapies, we need to clarify the index/ratio of ω-3 PUFAs in maternal and fetal fluids, delineate the ω-3 PUFA transport pathways, and establish the guidelines for FA profiling prepregnancy and during pregnancy-associated weight gain. Such therapies also need to take into account the gender of the fetus, and whether the patient is obese.

Genetic and Clinical Characteristics of Patients With Homozygous and Compound Heterozygous Familial Hypercholesterolemia From Three Different Populations: Case Series.

Homozygous familial hypercholesterolemia (HoFH) and compound heterozygous familial hypercholesterolemia (cHeFH) are rare disorders generated by disease-causing variants in both alleles of the LDLR or other familial hypercholesterolemia (FH)-related genes. HoFH and cHeFH are characterized by severely elevated low-density lipoprotein-cholesterol (LDL-C), frequently leading to early cardiovascular disease. We investigated the genetic and clinical characteristics of HoFH and cHeFH patients from the Slovenian FH registry and/or those who were previously diagnosed or managed at our institution (Slovenian, Pakhtun and Albanian ethnicity), where genetic testing is not available. Our study includes seven patients. Their median age at the time of clinical diagnosis was 6.3 years (2.9-12.9 years); 2/7 were females. Two patients were diagnosed through the universal FH screening and five patients were diagnosed due to the presence of xanthomas. All the mutations are present in LDLR gene: 7 different genotypes for HoFH (p.Cys167Leu, p.Asp178Asn, p.Cys243Tyr, p.Gly549Asp, p.Cys27Trp, p.Ile585Thr and p.Val797Met) and p.Gly549Asp/p.Gln384Pro genotype for cHeFH patient. The median initial level of LDL-C was 17.0 mmol/L [655 mg/dL] (range 7.6-21.6 mmol/L). The HoFH/cHeFH patients are clinically and genetically very diverse. The clinical criteria (as Simon Broome criteria) might be applicable already in children to raise suspicion of FH but in some cases fail to distinguish heterozygous FH and HoFH/cHeFH patients. However, genetic testing is helpful in confirming the diagnosis, also for a prompt awareness, better compliance to treatment and family screening.

Celiac Disease in Kosovar Albanian Children: Evaluation of Clinical Features and Diagnosis.

BACKGROUND: Celiac disease is an immune-mediated disorder characterized by variable clinical manifestations, specific antibodies, HLA-DQ2/DQ8 haplotypes, and enteropathy. OBJECTIVES: The aim of this study was to present the clinical spectrum and patterns of celiac disease in Kosovar Albanian children. METHODS: A cross-sectional retrospective study was performed with Albanian children aged 0-18 years, treated for celiac disease in the Pediatric Clinic, University Clinical Center of Kosovo from 2005 to 2016. RESULTS: During the study period, 63 children were treated for celiac disease. The mean age at diagnosis was 5.5 years (SD ± 3.31). The mean age at celiac disease onset was 3.3 years (SD ± 2.02), while the mean delay from the first symptoms indicative of celiac disease to diagnosis was 2.2 years (SD ± 2.09). More than 70% of the patients were diagnosed in the first 7 years of life, mainly presented with gastrointestinal symptoms, while primary school children and adolescents mostly showed atypical symptoms (p<0.001). The classical form of celiac disease occurred in 78% of the cases. Sixty (95%) patients carried HLA-DQ2.5, DQ2.2 and/or HLA-DQ8 heterodimers, and only three of them tested negative. CONCLUSION: Kosovo, as the majority of developing countries, is still facing the classical form of celiac disease as the dominant mode of presentation; as a result, most children with other forms of the celiac disease remain undiagnosed. Physicians should be aware of the wide range of clinical presentations and utilize low testing thresholds in order to prevent potential long-term problems associated with untreated celiac disease.

The Influence of Type 1 Diabetes Mellitus on Dental Caries and Salivary Composition.

Diabetes mellitus is the most common chronic disease that affects the oral health. The aim of the study is to evaluate the dental caries, salivary flow rate, buffer capacity, and Lactobacilli in saliva in children with type 1 diabetes mellitus compared to the control group. Methods. The sample consisted of 160 children of 10 to 15 years divided into two groups: 80 children with type 1 diabetes mellitus and 80 children as a control group. Dental caries was assessed using the DMFT index for permanent dentition. Stimulated saliva was collected among all children. Salivary flow rate and buffer capacity were measured, and the colonies of Lactobacillus in saliva were determined. The observed children have answered a number of questions related to their dental visits and parents' education. The data obtained from each group were compared statistically using the chi-square test and Mann-Whitney U-test. The significant level was set at p < 0.05. Results. DMFT in children with type 1 diabetes was significantly higher than that in the control group (p < 0.001). Diabetic children have a low level of stimulated salivary flow rate compared to control children (0.86 ± 0.16 and 1.10 ± 0.14). The buffer capacity showed statistically significant differences between children with type 1 diabetes and control group (p < 0.001). Also, children with type 1 diabetes had a higher count and a higher risk of Lactobacillus compared to the control group (p < 0.05 and p < 0.001). Conclusion. The findings we obtained showed that type 1 diabetes mellitus has an important part in children's oral health. It appears that children with type 1 diabetes are exposed to a higher risk for caries and oral health than nondiabetic children.

Novel Founder Mutation in FANCA Gene (c.3446_3449dupCCCT) Among Romani Patients from the Balkan Region.

BACKGROUND: Fanconi anemia is a rare autosomal recessive or X-linked disorder characterised by clinical and genetic heterogeneity. Most fanconi anemia patients harbour homozygous or double heterozygous mutations in the FANCA (60-65%), FANCC (10-15%), FANCG (~10%) or FANCD2 (3-6%) genes. We have already reported the FANCA variant c.190-256_283+1680del2040dupC as a founder mutation among Macedonian fanconi anemia patients of Gypsy-like ethnic origin. Here, we present a novel FANCA mutation in two patients from Macedonia and Kosovo. CASE REPORT: The novel FANCA mutation c.3446_3449dupCCCT was identified in two fanconi anemia patients with Romany ethnicity; a 2-year-old girl from Macedonia who is a compound heterozygote for a previously reported FANCA c.190-256_283+1680del2040dupC and the novel mutation and a 10-year-old girl from Kosovo who is a homozygote for the novel FANCA c.3446_3449dupCCCT mutation. The novel mutation is located in exon 35 in the FAAP20-binding domain which plays a crucial role in the FANCA-FAAP20 interaction and is required for integrity of the fanconi anemia pathway. CONCLUSION: The finding of the FANCA c.3446_3449dupCCCT mutation in two unrelated FA patients with Romani ethnicity from Macedonia and Kosovo suggests it is a founder mutation in the Romani population living in the Balkan region.

Dental Caries Among Kosovar Children with Type 1 Diabetes Mellitus

Objective: To evaluate the oral health status in children with type 1 diabetes mellitus. Material and Methods: Dental examinations, based on World Health Organization caries diagnostic criteria for DMFT index for permanent dentition and survey were performed among 160 children, aged 10-15-year-old, divided into two groups. The first group consisted of 80 children with type 1 diabetes mellitus (41 females, 39 males), and in the second group, consisted 80 healthy children (49 females, 31 males). Frequency, odds ratio and Mann-Whitney U test were used in the statistical analyses. The level of significance was set at 5%. Results: The higher mean of the DMFT index was found among children with type 1 diabetes compared to the healthy group. The mean DMFT index for diabetic children was 6.56 ± 3.56 and for the healthy group was 4.21 ± 2.63. Moreover, the frequency of decayed teeth was higher in children with type 1 diabetes than in the healthy group. The higher risk of caries was found in diabetic children compared with healthy for 1.35 times. A higher proportion of children, 61.25% with type 1 diabetes mellitus, reported that they brush their teeth once per day, 22.50% twice per day, and 16.25% rarely. From the healthy group, 46.25% of children brush their teeth once per day, and 42.50% twice per day and 11.25% rarely brush their teeth per day. Conclusion: Diabetic children are at higher risk for caries than are healthy children.

Prevalence of gestational diabetes mellitus according to IADPSG and NICE criteria.

OBJECTIVE: To investigate the impact of the International Association of Diabetic Pregnancy Study Group (IADPSG) diagnostic criteria on the prevalence of gestational diabetes mellitus (GDM) and overt diabetes as compared with the UK National Institute for Health and Care Excellence (NICE) criteria, and to evaluate the prevalence of maternal and perinatal outcomes among pregnant women with fasting plasma glucose (FPG) levels of 5.1-5.5 mmol/L. METHODS: A retrospective study was undertaken of data for women who underwent a 2-hour 75-g oral glucose tolerance test at 24-32 weeks of a singleton pregnancy at a center in Croatia between January 2012 and December 2014. RESULTS: Among 4646 included women, 1074 (23.1%) had GDM according to IADPSG criteria, 826 (17.8%) would be diagnosed according to NICE criteria, and 50 (1.1%) had overt diabetes. FPG levels were 5.1-5.5 mmol/L for 409 (8.8%) women. Compared with a control group (n=3391), these women had higher odds of large-for-gestational-age newborns (odds ratio 3.7, 95% CI 2.0-4.6) and cesarean delivery (odds ratio 1.8, 95% CI 1.3-2.3). CONCLUSION: Women with FPG levels of 5.1-5.5 mmol/L have an increased risk of adverse maternal and perinatal outcome, although they would not be diagnosed with GDM according to NICE criteria.

Hashimoto Thyroiditis and Nephrocalcinosis in a Child with Down Syndrome.

INTRODUCTION: Hypothyroidism has been reported to affect renal function and structure. However, the association of hypothyroidism with distal renal tubular acidosis (dRTA) is rarely reported in children. CASE PRESENTATION: We present a 6-year-boy with Down syndrome admitted in our department due to vomiting, weakness, polyuria, polydipsia, irritability and weight loss in the last few weeks. Investigations revealed features of hypokalemia, metabolic acidosis and alkaline urine consistent with dTRA. Abdominal ultrasound found nephrocalcinosis. In addition, Antithyroid peroxidase antibodies were positive, suggesting an autoimmune background for the pathogenesis of the tubular dysfunction. Treatment for dRTA and hypothyroidism was started and symptomatic improve was noticed. CONCLUSION: dRTA should be excluded in children with autoimmune disorders who develop weakness, polyuria, polydipsia or growth failure. Early diagnosis would reduce long-term complications.

Phenylketonuria screening and management in southeastern Europe - survey results from 11 countries.

BACKGROUND: We aimed to assess the current state of PKU screening and management in the region of southeastern Europe. METHODS: A survey was performed involving all identified professionals responsible for the PKU management in the 11 countries from South-Eastern region of Europe (Albania, Bulgaria, Bosnia and Herzegovina, Croatia, Kosovo, Macedonia, Moldova, Montenegro, Romania, Serbia, Slovenia). The questionnaire was designed to assess the characteristics regarding PKU management in three main areas: nation-wide characteristics, PKU screening, and characteristics of the PKU management in the responding centre. It consisted of 56 questions. The distribution and collection of the questionnaires (via e-mail) was taking place from December 2013 to March 2014. RESULTS: Responses from participants from 11 countries were included; the countries cumulative population is approx. 52.5 mio. PKU screening was not yet introduced in 4 of 11 countries. Reported PKU incidences ranged from 1/7325 to 1/39338 (and were not known for 5 countries). National PKU guidelines existed in 5 of 11 countries and 7 of 11 countries had PKU registry (registries included 40 to 194 patients). The number of PKU centers in each country varied from 1 to 6. Routine genetic diagnostics was reported in 4 of 11 countries. Most commonly used laboratory method to assess phenylalanine levels was fluorometric. Tetrahydrobiopterine was used in only 2 of 11 countries. Most frequently, pediatricians were caring for the patients. Dietitian was a member of PKU team in only 4 of 11 countries, while regular psychological assessments were performed in 6 of 11 countries. Patient's PKU society existed in 7 of 11 countries. CONCLUSIONS: The region of southeastern Europe was facing certain important challenges of PKU screening and management. Neonatal PKU screening should be introduced throughout the region. Furthermore, PKU management was falling behind internationally established standards-of-care in many aspects.

Newborn screening in southeastern Europe.

The aim of our study was to assess the current state of newborn screening (NBS) in the region of southeastern Europe, as an example of a developing region, focusing also on future plans. Responses were obtained from 11 countries. Phenylketonuria screening was not introduced in four of 11 countries, while congenital hypothyroidism screening was not introduced in three of them; extended NBS programs were non-existent. The primary challenges were identified. Implementation of NBS to developing countries worldwide should be considered as a priority.

Primary hypothyroidism and nipple hypoplasia in a girl with Wolcott-Rallison syndrome.

UNLABELLED: Wolcott-Rallison syndrome (WRS), caused by mutation in the EIF2AK3 gene encoding the PERK enzyme, is the most common cause of permanent neonatal diabetes mellitus (PNDM) in consanguineous families and isolated populations. Besides PNDM, it also includes skeletal abnormalities, liver and renal dysfunction, and other inconsistently present features. We present two siblings, who are WRS patients, and are Albanians from Kosovo born to unrelated parents. The older sister presented with PNDM, exocrine pancreatic insufficiency, short stature, microcephaly, normocytic anemia, delay in speech development, skeletal abnormalities, primary hypothyroidism, and hypoplastic nipples. Sequencing of the EIF2AK3 gene identified a homozygous mutation R902X in exon 13. The younger brother was diagnosed with PNDM and died from hepatic failure suggesting that he has been suffering from WRS as well. Including one previously reported patient from Kosovo carrying the same homozygous mutation, there are three WRS patients from this very small, ethnically homogenous region suggesting founder effect in this population. CONCLUSION: We postulate that thyroid hypoplasia with primary subclinical hypothyroidism already reported in two WRS patients and nipple hypoplasia could also be the phenotypic reflection of the mutation of pleiotropic EIF2AK3 gene in secretory cells.