The ethical framework for performing research with rare inherited neurometabolic disease patients.

The need for performing clinical trials to develop well-studied and appropriate medicines for inherited neurometabolic disease patients faces ethical concerns mainly raising from four aspects: the diseases are rare; include young and very young patients; the neurological impairment may compromise the capability to provide 'consent'; and the genetic nature of the disease leads to further ethical implications. This work is intended to identify the ethical provisions applicable to clinical research involving these patients and to evaluate if these cover the ethical issues. Three searches have been performed on the European regulatory/legal framework, the literature and European Union-funded projects. The European legal framework offers a number of ethical provisions ruling the clinical research on paediatric, rare, inherited diseases with neurological symptoms. In the literature, relevant publications deal with informed consent, newborn genetic screenings, gene therapy and rights/interests of research participants. Additional information raised from European projects on sharing patients' data from different countries, the need to fill the gap of the regulatory framework and to improve information to stakeholders and patients/families. CONCLUSION: Several recommendations and guidelines on ethical aspects are applicable to the inherited neurometabolic disease research in Europe, even though they suffer from the lack of a common ethical approach. What is Known: • When planning and conducting clinical trials, sponsors and researchers know that clinical trials are to be performed according to well-established ethical rules, and patients should be aware about their rights. • In the cases of paediatric patients, vulnerable patients unable to provide consent, genetic diseases' further rules apply. What is New: • This work discusses which ethical rules apply to ensure protection of patient's rights if all the above-mentioned features coexist. • This work shows available data and information on how these rules have been applied.

Review and evaluation of the methodological quality of the existing guidelines and recommendations for inherited neurometabolic disorders.

BACKGROUND: Inherited neurometabolic disorders (iNMDs) represent a group of almost seven hundred rare diseases whose common manifestations are clinical neurologic or cognitive symptoms that can appear at any time, in the first months/years of age or even later in adulthood. Early diagnosis and timely treatments are often pivotal for the favorable course of the disease. Thus, the elaboration of new evidence-based recommendations for iNMD diagnosis and management is increasingly requested by health care professionals and patients, even though the methodological quality of existing guidelines is largely unclear. InNerMeD-I-Network is the first European network on iNMDs that was created with the aim of sharing and increasing validated information about diagnosis and management of neurometabolic disorders. One of the goals of the project was to determine the number and the methodological quality of existing guidelines and recommendations for iNMDs. METHODS: We performed a systematic search on PubMed, the National Guideline Clearinghouse (NGC), the Guidelines International Network (G-I-N), the Scottish Intercollegiate Guideline Network (SIGN) and the National Institute for Health and Care Excellence (NICE) to identify all the published guidelines and recommendations for iNMDs from January 2000 to June 2015. The methodological quality of the selected documents was determined using the AGREE II instrument, an appraisal tool composed of 6 domains covering 23 key items. RESULTS: A total of 55 records met the inclusion criteria, 11 % were about groups of disorders, whereas the majority encompassed only one disorder. Lysosomal disorders, and in particular Fabry, Gaucher disease and mucopolysaccharidoses where the most studied. The overall methodological quality of the recommendation was acceptable and increased over time, with 25 % of the identified guidelines strongly recommended by the appraisers, 64 % recommended, and 11 % not recommended. However, heterogeneity in the obtained scores for each domain was observed among documents covering different groups of disorders and some domains like 'stakeholder involvement' and 'applicability' were generally scarcely addressed. CONCLUSIONS: Greater efforts should be devoted to improve the methodological quality of guidelines and recommendations for iNMDs and AGREE II instrument seems advisable for new guideline development. The elaboration of new guidelines encompassing still uncovered disorders is badly needed.

Mitochondrial myopathy associated with a novel 5522G>A mutation in the mitochondrial tRNA(Trp) gene.

We report a novel pathogenic mutation of the mitochondrial transfer RNA (tRNA) gene for tryptophan in a patient with isolated myopathy and persistently elevated creatine kinase. Muscle studies revealed ragged red fibres and decreased activity of respiratory chain complex I and cytochrome c oxidase (COX). Sequencing of the 22 mitochondrial tRNA genes revealed a mutation m.5522G>A, which alters a conserved base pairing in the D-stem of the tRNA for tryptophan. The mutation was heteroplasmic with a mutational load between 88 and 99% in COX-negative fibres. This case contributes to the genetic heterogeneity of mitochondrial diseases caused by mutations in mitochondrial tRNA genes.

Social and medical care of preschool children with epilepsy in Croatia: population-based survey.

BACKGROUND: Early detection of mental retardation and other epilepsy-associated impairments is essential for successful medical and social care of children with epilepsy; the corresponding information for children in Croatia has not yet been known. AIMS OF THE STUDY: To obtain the basic information of epilepsy-associated disability in preschool children, and fundamentals of their medical and social care. METHODS: Data about mental retardation and other associated impairments (motor, speech, seeing, hearing), antiepileptic drug therapy and diurnal residence were collected by means of questionnaires completed by physicians working in primary health care (PHPs). Only children (0-7 years) with active epilepsy confirmed previously by neuropaediatricians were included. RESULTS: A total of 37 PHPs provided the required data for 116 children. One or more impairments were found in 56% children; most frequent were motor impairments (47%), speech impairments (42%) and mental retardation (40%). The regular kindergarten attendance rate of children without impairment (33%) was not different from the children without epilepsy, but high proportion (76%) of children with impairment stayed with their families during weekdays. In this subgroup monotherapy was more rarely used (64% vs. 90% in children without impairment (p<0.01)). Valproate was predominantly used (56%) in children with and without impairment; lamotrigine was more frequently used in the former subgroup (p<0.01). CONCLUSIONS: Existence of associated impairments has significant impact on medical and social care in preschool children with epilepsy. These children need an early diagnosis and consecutive multidisciplinary care of their intellectual and body impairments, as well as problems in social development.

Age-related pattern of the antiepileptic drug utilization in active epilepsy: a population-based survey.

The aim of this study was to investigate the relationship between antiepileptic drug (AED) utilization and patient age in a population of patients treated by primary health care physicians. Data were collected by using questionnaires completed by family physicians and paediatricians working in primary health care. Only patients with active epilepsy confirmed previously by neurologists or neuropaediatricans were included. One hundred and twenty-three physicians provided the requested data for 966 patients (range 1-92 years). Most frequently prescribed AEDs were barbiturates (BARB) (37%) and carbamazepine (CBZ) (37%). Valproic acid derivates (VPA) were prescribed in 28%, but the rate was higher (51%) in children. By calculating the correlation between age and the prescription of single AEDs across the whole sample, linear correlations were found for BARB (r = 0.94; p < 0.01), VPA (r = -0.93; p < 0.01) and for topiramate (TPM) (r = -0.90; p < 0.01). Since our results showed significant correlations between age and the use of the majority of AEDs, we concluded that the age may be considered a methodological bias in the presentation of data. Therefore we calculated AED utilization as the age-adjusted prevalence rates (per/1000 inhabitants). For the most commonly prescribed AEDs they were: BARB 1.8 (95% CI 1.6-2.0), CBZ 1.9 (95% CI 1.7-2.1), VPA 1.3 (95% CI 1.1-1.5), lamotrigine (LTG) 0.7 (CI 95% 0.6-0.8), TPM 0.6 (CI 95% 0.5-0.7). In conclusion, the age of patients has a significant impact on the prescription patterns not only between children and adults, but at every age. Therefore we suggest that reporting of AED utilization pattern should also include age-standardized prevalence rates of individual AED utilization.

Prospective surveillance of Croatian pregnant women on lamotrigine monotherapy--aspects of pre-pregnancy counseling and drug monitoring.

We prospectively surveyed 23 pregnant women with epilepsy on lamotrigine monotherapy and reported outcome of their pregnancies, including one fetal intrauterine death, one spontaneous abortion and two preterm deliveries. There were no congenital malformations in their offspring. Women with pregnancy planning and folic acid intake delivered babies with higher values of birth weight and birth length. There was large inter-patient variation during drug monitoring and in the need of dose adjustment. Individual approach to every woman and monotherapy with minimal effective lamotrigine dose with frequent drug monitoring enhances the possibility for successful pregnancy. The management of women with epilepsy should begin with pre-pregnancy counseling. Planned pregnancy enables periconceptional folic acid supplementation. Despite the small number of cases, these data indicate that la motrigine treatment during pregnancy might be relatively safe. Larger prospective studies are needed to obtain adequate power for statistical analysis including long-term cohort studies.

Diagnostic and therapeutic doubts in retrobulbar neuritis in children.

Retrobulbar neuritis is often very complicated clinical entity. The most common cause of retrobulbar neuritis is demyelinating disease of CNS. This report is to express some other uncommon causes of it. Three children, age 8 to 12 with sudden and severe visual loss are presented. The diagnosis of retrobulbar neuritis is made by complete ophtalmological examination in consultation with neuropediatrics and neuroradiologist. Different ethiological causes of retrobulbar neuritis are found: pranasal sinusitis, functional visual loss and pseudotumor cerebri. In first two children complete therapeutically effort was as expected, and by child with pseudotumor cerebri there was no improvement of visual acuity, even after 6 months. In this presentation the authors want to emphasise some uncommon causes of retrobulbar neuritis.

[Treatment of West syndrome]. / Lijecenje Westovog sindroma.

PURPOSE: West syndrome (WS) is one of the catastrophic epileptic syndromes in infancy characterized by a triad of infantile spasms, psychomotor deterioration and hypsarrhythmic EEG pattern. WS is commonly associated with poor long-term outcome, especially in symptomatic cases, with development of other seizure types, impaired cognitive and psychosocial functioning. The aim of our study was to evaluate the efficacy of the control of infantile spasms using synthetic ACTH or vigabatrin in newly diagnosed cases and to correlate it with the underlyning causes, outcome and adverse effects. PATIENTS AND METHODS: The database of children with WS seen at the Neuropediatric Unit and followed at outpatient clinics from January 1, 1994 until December 31, 2003 were reviewed. The diagnosis of WS following the criteria of ILAE was made in 32 patients. RESULTS: Data were collected for 32 children (9 girls and 23 boys). According to the etiology, 5 (15.6%) were cryptogenic, and 1 (3.1%) was idiopathic. In 26 (81.2%) symptomatic cases, hypoxic-ischemic encephalopathy (69.2%) was the most common etiologic factor, followed by central nervous system anomaly including malformation of cortical development (11.5%), and Sturge Weber syndrome (3.8%), and chromosomal translocation with Down syndrome (11.5%). In 65.1% of symptomatic cases birth occurred prematurely. The mean age at spasm onset was 5.8 months, and mean age at diagnosis and treatment 7.2 months. Between 1994 and 1996 synthetic ACTH was used for treatment of WS in 7 patients (1 cryptogenic and 6 symptomatic), spasm control was achieved in 6, hypsarrhythmia disappeared in 5, and vigabatrin was added after synthetic ACTH in 3 patients. In one child synthetic ACTH was stopped because of arterial hypertension. All children had Cushing syndrome. After 1996, vigabatrin was administrated to 5 children with cryptogenic and 20 children with symptomatic WS. In 22/32 spasm control was achieved within 15 days. Synthetic ACTH was added in 3 children with spasms and hypsarrhythmia disappeared in 1 child. There was no recurrence of WS. The mean follow-up in 27 children was 4.6 (0.5 to 9.9 years) whereas 5 were lost from follow-up. Of 6/27 children with cryptogenic WS, 1 had idiopathic WS, 3 had normal psychomotor development and 2 had psychomotor retardation, without epileptic fits and still receiving AED. Of 21/27 children with symptomatic WS 76.2% had severe psychomotor retardation, 42.8% had epilepsy, 23.8% had intractable epileptic fits, and 2 children with Down syndrome were without epilepsy and without AED. Lennox-Gastaut syndrome developed in 14.2% (3/21 children); 1 of them died at the age of 3.5 years from acute gastric bleeding during the administration of synthetic ACTH, and an other child died at the age of 5.5 years from infection and respiratory insufficiency. The mortality rate was 7.4% (2/27 children). DISCUSSION AND CONCLUSION: The cryptogenic etiology is associated with a very low risk of poor outcome in WS. In children with normal development and regular school performance an idiopathic etiology can be presumed. The children with Down syndrome had a relatively benign outcome with regard to seizure control compared with symptomatic infantile spasms in the general population. In symptomatic WS caused by hypoxic-ischemic encephalopathy the outcome was linked with coexistence of other forms of epilepsy and neurologic deficit. The poor prognosis concerning intractable nature of the seizures and serious neurologic deficit is recorded in children with malformation of cortical development and Sturge Weber syndrome. The outcome of these children is determined by the brain damage other than by epilepsy itself. Regarding the treatment with synthetic ACTH or vigabatrin, the control of WS was the same for cryptogenic and symptomatic forms, one drug may be effective if the other drug fails. Synthetic ACTH can have many side effects, even death. The visual field defect is associated with vigabatrin, but can be avoided with careful funduscopic follow-up. Vigabatrin can be suggested as the first drug for WS; if spasms persist after 15 days with a dose of 150 mg/kg, synthetic ACTH should be considered.

Increase of classical antiepileptic drug utilization in Croatia during the process of introducing the new generation of drugs.

During the last decade the process of introducing the new generation of antiepileptic drugs (AEDs) has substantially changed the ways of treating epilepsy. Although a great deal of information about the role of new drugs has been accumulated, much less attention was paid to the impact of the new generation of AEDs on the utilization of classical AEDs. In order to detect the relation between the new and classical AEDs, the data about drug consumption in Croatia in the period 2000-2002 were analyzed. The main results indicated that the growth utilization rate (15%) was more the result of increasing consumption of the classical antiepileptic substances (in almost 2/3). It has been discussed that one of the possible interpretations for this phenomenon could lie in the fact that the continuing process of introducing the new AEDs was accompanied by a great number of educational activities. These activities have led to greater awareness of the facilities in treating epilepsy and consequently to a more active therapeutic approach, which encompassed both generations of drugs, even more the older one.

Carnitine membrane transporter deficiency: a long-term follow up and OCTN2 mutation in the first documented case of primary carnitine deficiency.

Three older patients were diagnosed with systemic carnitine deficiency in childhood nearly a generation ago and have together been treated for more than 50 patient years. Treatment improved tissue carnitine stores (proven in two) and eliminated most of the signs and symptoms of carnitine deficiency. All three have continued to respond to carnitine therapy and remain well except for the irreversible sequelae of the pretreatment illnesses. We demonstrate here that transformed lymphocytes from the first documented case of plasma membrane carnitine transporter deficiency fail to take up carnitine from the medium. The analysis of the cDNA of this patient and his parents revealed a homozygous frameshift mutation, 1027delT in exon 4. The resulting polypeptide terminates after amino acid 295. His parents are heterozygous for this mutation. The deletion resulted in predominately abnormal mRNA splicing with either a 13 or 19bp insertion between the junction of exons 3 and 4. The 13/19bp insertions were found in both parents, predominantly in cis with the deletion, and rarely seen with normal alleles from either parents or controls.

Novel OCTN2 mutations: no genotype-phenotype correlations: early carnitine therapy prevents cardiomyopathy.

Primary systemic carnitine deficiency or carnitine uptake defect (OMIM 212140) is a potentially lethal, autosomal recessive disorder characterized by progressive infantile-onset cardiomyopathy, weakness, and recurrent hypoglycemic hypoketotic encephalopathy, which is highly responsive to L-carnitine therapy. Molecular analysis of the SLC22A5 (OCTN2) gene, encoding the high-affinity carnitine transporter, was done in 11 affected individuals by direct nucleotide sequencing of polymerase chain reaction products from all 10 exons. Carnitine uptake (at Km of 5 microM) in cultured skin fibroblasts ranged from 1% to 20% of normal controls. Eleven mutations (delF23, N32S, and one 11-bp duplication in exon 1; R169W in exon 3; a donor splice mutation [IVS3+1 G > A] in intron 3; frameshift mutations in exons 5 and 6; Y401X in exon 7; T440M, T468R and S470F in exon 8) are described. There was no correlation between residual uptake and severity of clinical presentation, suggesting that the wide phenotypic variability is likely related to exogenous stressors exacerbating carnitine deficiency. Most importantly, strict compliance with carnitine from birth appears to prevent the phenotype.