ObjectiveThis study aimed to assess the diagnostic yield, clinical indications, and utility of next-generation sequencing (NGS) testing since its implementation through collaboration between the University of Rijeka Faculty of Medicine and the Clinical Hospital Centre Rijeka.Materials and MethodsThis retrospective study included patients referred between 2018 and 2023 from the Clinical Hospital Centre Rijeka to the University of Rijeka Faculty of Medicine for genetic testing, primarily using exome sequencing.ResultsBetween April 2018 and December 2023, 412 patients were referred for exome sequencing, of whom 353 (85.7%) underwent diagnostic genetic testing. A notable increase in tests ordered was observed over time. Patients were most frequently referred from Pediatrics (55.0%), Neurology (29.5%), Cardiology (7.4%), Ophthalmology (3.4%), and others (4.7%). A diagnosis was confirmed in 103/353 patients, corresponding to an overall diagnostic yield of 29.2%, and an adjusted diagnostic yield of 27.2% after collapsing related individuals into single family units. In these confirmed cases, 83 distinct disorders involving 71 unique genes were identified, with most patients showing heterozygous variants and several recurrent disorders and genes. Variants of uncertain significance were reported in 35/353 (9.9%) patients.ConclusionThe 27.2% diagnostic yield demonstrates effective integration of NGS into tertiary clinical practice. The recent introduction of medical genetics specialization is expected to further improve referral quality, variant interpretation, and overall diagnostic outcomes.

The Exposure and Health Examination Survey (EXHES) cohort aims to elucidate the impact of environmental exposures (the external exposome) and their biological markers (the internal exposome) on childhood health conditions, asthma and allergies, obesity, and cognitive development in particular. Utilizing singletons and twins helped differentiate environmental effects from genetic influences due to the shared genetic background in twins. The EXHES cohort includes 2356 mother-child pairs across 10 European countries, comprising 1945 singletons and 411 twins, with data collected during the crucial first 1000 days of life. Data were gathered through epidemiological questionnaires and biomarkers, including blood, urine, hair, and breast milk from mothers, and cord blood, placenta, and cord tissues from children. Findings confirm that twin pregnancies are linked with increased risks of pregnancy complications, preterm birth, cesarean delivery, low birthweight, maternal health problems during pregnancy and a lower risk of macrosomia. Moreover, mothers of twins were more likely to have asthma, while higher maternal education was associated with a lower likelihood of twin births. The EXHES cohort provides a robust framework to be adopted in other studies for comparing singletons and twins to better understand how the exposome affects early child development and health outcomes. This approach offers new insights into the interplay between environmental and biological factors in shaping long-term health.

Terapijska primjena ketogene dijete postala je moćnim terapijskim pristupom u liječenju tvrdokornih epilepsija, osobito u dojenčadi i djece. Povezivanje praksi utemeljenih na dokazima i kliničke stručnosti dovelo je do razvoja internacionalnih sveobuhvatnih smjernica s ciljem standardizacije primjene i optimizacije učinkovitosti ketogene dijete na temelju kojih su pripremljene nacionalne smjernice. Ove smjernice pružaju strukturirani protokol za zdravstvene djelatnike koji primjenjuju ketogene dijete u svojoj kliničkoj praksi. Pored toga, naglašavaju važnost individualiziranih planova liječenja, pažljive procjene oboljelih i stalnog praćenja kako bi se osigurala sigurnost i učinkovitost. Središnji aspekt istaknut u ovim smjernicama jest temeljita procjena prikladnosti terapijske primjene ketogene dijete za oboljelog, uzimajući u obzir čimbenike poput dobi, epileptičkog sindroma, učestalosti napadaja, komorbiditeta i osobne preferencije oboljelog/obitelji. Uvođenje i održavanje primjene ketogene dijete zahtijeva blisku suradnju multidisciplinarnog tima, uključujući neuropedijatre, nutricioniste, roditelje/ skrbnike i druge zdravstvene djelatnike kako bi se lakše upravljalo mogućim izazovima i optimizirali ishodi dijetoterapije. Ključne komponente smjernica obuhvaćaju protokole za preevaluaciju, uvođenje i praćenje. Fokus je na postizanju i održavanju odgovarajuće nutritivne ketoze uz osiguravanje adekvatnosti prehrane i umanjivanje nuspojava. Redovito praćenje biokemijskih parametara, parametara rasta i kontrole epileptičkih napadaja imperativ je za podešavanje ketogene dijete i učinkovito ublažavanje mogućih neželjenih pojava. Nadalje, smjernice zagovaraju kontinuirano obrazovanje i podršku oboljelima i roditeljima/skrbnicima kako bi se poboljšalo pridržavanje dijetoterapije i promicao dugoročni uspjeh. Ove smjernice služe kao važan izvor za zdravstvene djelatnike, nudeći preporuke utemeljene na dokazima i praktične uvide za upravljanje izazovima terapijske primjene ketegene dijete, u konačnici nastojeći poboljšati njen učinak i kvalitetu života oboljelih od tvrdokornih epilepsija.

Lead (Pb) is a global contaminant associated with multiple adverse health effects. Humans are especially vulnerable during critical developmental stages. During pregnancy, exposure to Pb can occur through diet and release from maternal bones. Apolipoprotein E gene (APOE) variants (ɛ2, ɛ3, ɛ4 alleles) may influence sex steroid hormones, bone metabolism, and Pb kinetics.We examined the interplay among maternal APOE (mAPOE) genotypes, fetal sex, parity, and Pb in maternal and cord blood (mB-Pb, CB-Pb) using linear regression models. Our study involved 817 pregnant women and 772 newborns with measured adequate levels of zinc and selenium. We compared carriers of the ε2 and ε4 alleles to those with the ε3/ε3 genotype.The geometric means (range) of mB-Pb and CB-Pb were 11.1 (3.58–87.6) and 9.31 (1.82–47.0) ng/g, respectively. In cases with female fetuses, the maternal mAPOE ε2 allele was associated with higher, while the mAPOE ε4 allele was associated with lower mB-Pb and CB-Pb levels. Nulliparity increased the strength of the observed associations. These findings highlight the significance of mAPOE genetics, fetal sex, and parity in prenatal Pb kinetics. Notably, the maternal ε2 allele may increase the risk of Pb exposure.

Abstract Due to the increasing importance of exposome in environmental epidemiology, feasibility and usefulness of an Environmental Data Management System (EDMS) using Open Data was evaluated. The EDMS includes data from 10 European cities (Celje (Slovenia), Łódź (Poland), Manchester (UK), Palermo (Italy), Paris (France), Porto (Portugal), Regensburg (Germany), Reus (Spain), Rijeka (Croatia), Thessaloniki (Greece)) about external non-specific and specific exposome factors at the city or country level (2017–2020). Findings showed that the highest values of life expectancy were in Reus females (86 years) and Palermo males (81 years). UK had the highest obesity rate (28%), Croatia the highest prescribed drug consumption (62%), Greece and Portugal the highest smoking rates (37%, 42%) and daily alcohol consumption (21%), respectively. The most polluted cities were Thessaloniki for PM10 (38 µg/m3), Łódź for PM2.5 (25 µg/m3), Porto for NO2 (62 µg/m3) and Rijeka for O3 (92 µg/m3). Thessaloniki had the highest grey space (98%) and Łódź the highest cumulative amount of pollen (39,041 p/m3). The highest daily noise levels ≥ 55 dB was in Reus (81% to traffic) and Regensburg (21% to railway). In drinking water, arsenic had the highest value in Thessaloniki (6.4 µg/L), boron in Celje (24 mg/L) and lead in Paris (46.7 µg/L). Portugal and Greece showed the highest pesticide residues in food (7%). In conclusion, utilizing open-access databases enables the translation of research findings into actionable strategies for public health interventions.

Background: Late infantile neuronal ceroid lipofuscinosis type 2 (CLN2) is a rare neurodegenerative disease that generally appears in children between 2 and 4 years old, leading to seizures and a progressive loss of language and motor functions. As the disease progresses, affected individuals typically experience blindness and ultimately pass away in late childhood. Treatment with intracerebroventricular cerliponase alfa has been shown to slow the deterioration of motor and language functions compared to the natural progression of the disease. We aim to highlight the early symptoms of CLN2 which help with early diagnosis and timely treatment initiation in children with specific medical indications, as well as identify medical contraindications for enzyme replacement therapy. Methods: We describe five Croatian patients and one Bosnia and Herzegovinian patient with CLN2 disease, analyzing the clinical characteristics, neuroimaging findings, electroencephalogram results, genetic analysis, treatment indications and contraindications, and disease progression. Results: All six patients presented with seizures: focal seizures (n = 1), myoclonic–atonic seizures (n = 1), febrile seizures (n = 2), and tonic–clonic seizures (n = 2), along with language delay (n = 6). Despite this, one patient refused treatment, two were initially included in the clinical trial and then continued treatment, one did not indicate starting treatment, and three continued treatment. One patient, after 4.5 years of treatment, no longer had medical indications for the therapy, which was discontinued. The other two patients who received treatment had a significant slowing of disease progression. Conclusions: The early onset of seizures between ages 2 and 4, alongside delayed language development, is a defining characteristic of CLN2 disease. Enzyme replacement therapy using cerliponase alfa represents the initial treatment for neuronal ceroid lipofuscinosis type 2, targeting the underlying cause of the disease. It effectively delays the progression of language and motor decline in patients diagnosed with this condition.

Febrilne konvulzije najčešći su konvulzivni poremećaj u djetinjstvu, zbog čega bi liječnici trebali biti upoznati s pravilnom procjenom i liječenjem ovoga uobičajenog stanja. Definira se kao bilo koji napadaj popraćen vrućicom, bez infekcije središnjega živčanog sustava, koji se javlja kod djece između šest mjeseci i pet godina. U upotrebi je pet kriterija prema kojima se klasificiraju kao jednostavne ili složene febrilne konvulzije. Ova klasifikacija nudi drugačije smjernice za kliničku praksu, poglavito odluku o provođenju lumbalne punkcije kako bi se isključila intrakranijalna infekcija. Hrvatsko društvo za dječju neurologiju izradilo je 2012. godine prve smjernice za racionalnu dijagnostiku i terapiju febrilnih konvulzija koje su trebale biti izvedive u svim bolničkim ustanovama u Republici Hrvatskoj. Iako su smjernice bile od velike pomoći mnogim kliničarima, dobivanjem novih medicinskih dokaza i razumijevanjem kliničkih entiteta koju se prezentiraju febrilnim konvulzijama, a variraju od samolimitirajućeg poremećaja bez dugoročnih posljedica preko samoograničavajućih i medikamentima kontroliranih entiteta pa sve do teških epileptičkih encefalopatija, nove smjernice pridonose postavljanju dijagnoze u hitnoj ambulanti, uključuju preporuke o hospitalizaciji, primjeni elektroencefalografije, neuroslikovnih pretraga, profilaksi diazepamom i antipireticima. Iako nove smjernice sadrže nove kliničke preporuke, ostavljaju neka neriješena pitanja na koja buduća klinička istraživanja trebaju ponuditi odgovore.

Rano suzbijanje, prekidanje epileptičkih napadaja središnji je stup u strategiji njihova liječenja. Većina epileptičkih napadaja događa se izvan medicinske ustanove, najčešće u kućnom okruženju. Stoga smo izradili i predložili ovaj algoritam zbrinjavanja motoričkih/konvulzivnih epileptičkih napadaja u izvanbolničkom okruženju, prvenstveno s ciljem pružanja uniformne informacije i edukacije roditelja/skrbnika te za olakšavanje snalaženja liječnicima primarne zdravstvene zaštite i timovima hitne medicinske pomoći. Za potrebe pisanja ovog rada analizirali smo i sintetizirali postojeće smjernice u cilju preporuke jednostavnog, razumljivog i racionalnog terapijskog algoritma koji savjetujemo i primjenjujemo kao službeni algoritam preporuka naše ustanove. U njegovoj izradi usmjerili smo se na svjetske i nacionalne preporučene algoritme kao i dostupne studije koje su, koliko je to moguće, temeljene na dokazima. Prema našim saznanjima ovo je prvi pisani nacionalni algoritam takve vrste te do sada nisu postajale pisane smjernice/upute za liječenje epileptičkih napadaja, produljenih epileptičkih napadaja i epileptičkog statusa u izvanbolničkom okruženju uzimajući u obzir dostupnost, racionalnost i primjenjivost pojedinih lijekova u izvanbolničkim uvjetima, uvažavajući kliničku praksu u Republici Hrvatskoj. Sukladno navedenom, primjena benzodiazepina (BZD), vremenski jasno definirana, smatra se prvom linijom liječenja epileptičkog napadaja. Izvanbolnička primjena BZD-a, prvenstveno midazolama za orkomukuznu primjenu ili diazepama za rektalnu primjenu, povezana je s kraćim trajanjem generaliziranih konvulzivnih napadaja, smanjenjem vjerojatnosti ponavljajućih napadaja i smanjenjem broja posjeta hitnoj pomoći. Ukoliko napadaj potraje >3 minuta, svakako je potrebno žurno primijeniti jedan od BZD-a, pod pretpostavkom da ga se posjeduje (midazolam oromukozno – prednost! ili diazepam rektalno), te isto ponoviti u slučaju daljnjeg trajanja napadaja duljeg od pet minuta te pozvati/alarmirati sustav hitne medicinske pomoći. Uz provođenje mjera održavanja vitalnih funkcija potrebno je inzistirati na intravaskularnom pristupu za ponovljenu primjenu BZD-a, no doza BZD-a može se ponoviti intramuskularno. U rijetkim slučajevima može se primijeniti fenobarbiton intramuskularno, kao i levetiracetam intravenozno, a u slučaju izostanka njihova učinka započeti s primjenom midazolama u trajnoj infuziji. Dostupnost primjerenih oblika BZD-a, sukladno smjernicama, u izvanbolničkom okruženju kao i njihova pravilna uporaba – pravodobna primjena i odgovarajuće doziranje početkom napadaja – dva su ključna koraka prema poboljšanju cjelokupne zdravstvene skrbi u djece s visokim rizikom za pojavu epileptičkih napadaja i djece s epilepsijom. Navedeno ujedno predstavlja i pravovremeno suzbijanje produljenih epileptičkih napadaja i prevenciju epileptičkog statusa.

Epilepsy is one of the most common neurological disorders with diverse phenotypic characteristics and high genetic heterogeneity. Epilepsy often occurs in childhood, so timely diagnosis and adequate therapy are crucial for preserving quality of life and unhindered development of a child. Next-generation-sequencing (NGS)-based tools have shown potential in increasing diagnostic yield. The primary objective of this study was to evaluate the impact of genetic testing and to investigate the diagnostic utility of targeted gene panel sequencing. This retrospective cohort study included 277 patients aged 6 months to 17 years undergoing NGS with an epilepsy panel covering 142 genes. Of 118 variants detected, 38 (32.2%) were not described in the literature. We identified 64 pathogenic or likely pathogenic variants with an overall diagnostic yield of 23.1%. We showed a significantly higher diagnostic yield in patients with developmental delay (28.9%). Furthermore, we showed that patients with variants reported as pathogenic presented with seizures at a younger age, which led to the conclusion that such children should be included in genomic diagnostic procedures as soon as possible to achieve a correct diagnosis in a timely manner, potentially leading to better treatment and avoidance of unnecessary procedures. Describing and discovering the genetic background of the disease not only leads to a better understanding of the mechanisms of the disorder but also opens the possibility of more precise and individualized treatment based on stratified medicine.

Introduction Neuronal ceroid lipofuscinosis type 2 (CLN2 disease) is a rare pediatric neurodegenerative condition, which is usually fatal by mid-adolescence. Seizures are one of the most common early symptoms of CLN2 disease, but patients often experience language deficits, movement disorders, and behavioral problems. Diagnosis of CLN2 disease is challenging (particularly when differentiating between early-onset developmental, metabolic, or epileptic syndromes), and diagnostic delays often overlap with rapid disease progression. An enzyme replacement therapy (cerliponase alfa) is now available, adding CLN2 disease to the list of potentially treatable disorders requiring a prompt diagnosis. Areas covered Although advances in enzymatic activity testing and genetic testing have facilitated diagnoses of CLN2 disease, our review highlights the presenting symptoms that are vital in directing clinicians to perform appropriate tests or seek expert opinion. We also describe common diagnostic challenges and some potential misdiagnoses that may occur during differential diagnosis. Expert opinion An awareness of CLN2 disease as a potentially treatable disorder and increased understanding of the key presenting symptoms can support selection of appropriate tests and prompt diagnosis. The available enzyme replacement therapy heralds an even greater imperative for early diagnosis, and for clinicians to direct patients to appropriate diagnostic pathways.

Epilepsy is one of the most common chronic diseases in children, and cannot be controlled with conventional antiepileptic drugs in 30% of cases. Therefore, in these cases, alternative approach such as corticosteroid therapy (CT) is used. The aim of this study was to analyze different types of CT used to treat drug-resistant childhood epilepsies, treated at Rijeka University Hospital Centre during a 5-year period (2016-2020). This retrospective study included 32 patients. The following parameters were analyzed: number of patients with a particular diagnosis, average age (in months) at the onset of epilepsy, average epilepsy duration (in months) prior to CT, average number of antiepileptic drugs used prior to CT, presence of changes on magnetic resonance imaging (MRI), presence of comorbidities, and types of CT. The average age at the onset of epilepsy was 14 months and average epilepsy duration prior to CT was 16 months. On average, 5 antiepileptic drugs were used prior to CT. MRI changes were present in 53.13% and comorbidities in 81.25% of study patients. Prednisone therapy was used in 28.13%, combined therapy with prednisone and methylprednisolone in 65.63%, and methylprednisolone in 6.25% of patients. Study results revealed the use of CT for particular diagnosis to differ among the centers, as well as within the same center, so it is important to highlight the importance of reaching universal guidelines for CT therapy of childhood epilepsies.

Few studies provide a detailed description of dietary habits during pregnancy, despite the central role of nutrition for the health of the mother and offspring. This paper describes the dietary habits, energy and nutrient intake in pregnant women from four countries belonging to the Mediterranean PHIME cohort (Croatia, Greece, Italy and Slovenia) and evaluates their adherence to the European Food Safety Authority (EFSA) recommendations. A total of 1436 women were included in the present analysis. Maternal diet was assessed using a food frequency questionnaire (FFQ). The mean macro and micronutrient intakes were estimated and compared with the dietary reference values (DRVs). The percentage distribution of the 16 food groups in the total intake of each macronutrient was estimated. All women shared a similar diet during pregnancy; almost all the women in the four countries exceeded the DRV for sugars, and the total fat intake was above the DRV in most women in all the countries, as was the contribution of saturated fatty acids (SFAs) to the total energy intake. In all four countries, we observed an increased risk of micronutrient deficiency for iron, folate and vitamin D. Shared guidelines, implemented at both the national and European level, are essential to improve the maternal nutritional status during pregnancy.

We explored the impact of coronavirus virus 2019 (COVID‐19) pandemic on patients with Dravet syndrome (DS) and their family. With European patient advocacy groups (PAGs), we developed an online survey in 10 languages to question health status, behavior, personal protection, and health services before and after lockdown. Approximately 538 European PAG members received electronic invitations. Survey ran from April 14, to May 17, 2020, with 219 answers; median age 9 year 10 months. Protection against infection was highly used prior to COVID‐19, but 88% added facemask‐use according to pandemic recommendations. Only one patient was tested positive for COVID‐19. Most had stable epilepsy during lockdown, and few families (4%) needed emergency care during lockdown. However, behavior disorder worsened in over one‐third of patients, regardless of epilepsy changes. Half of appointments scheduled prior to lockdown were postponed; 12 patients (11%) had appointments fulfilled; and 39 (36%) had remote consultations. Responders welcomed remote consultations. Half of responders were unsatisfied with psychological remote support as only few (21 families) received this support. None of the five of patient in clinical trials stopped investigational treatment. Prior adoption of protective measures against general infection might have contributed to avoiding COVID‐19 infections. Protocols for the favored remote contact ought to now be prepared.

Glucose transporter type 1 (GLUT1) is the most important energy carrier of the brain across the blood–brain barrier, and a genetic defect of GLUT1 is known as GLUT1 deficiency syndrome (GLUT1DS). It is characterized by early infantile seizures, developmental delay, microcephaly, ataxia, and various paroxysmal neurological phenomena. In most cases, GLUT1DS is caused by heterozygous single-nucleotide variants (SNVs) in the SLC2A1 gene that provoke complete or severe impairment of the functionality and/or expression of GLUT1 in the brain. Despite the rarity of these diseases, GLUT1DS is of high clinical interest since a very effective therapy, the ketogenic diet, can improve or reverse symptoms, especially if it is started as early as possible. We present a clinical phenotype, biochemical analysis, electroencephalographic and neuropsychological features of an 11-month-old boy with myoclonic seizures, hypogammaglobulinemia, and mildly impaired gross motor development. Using sequence analysis and deletion/duplication testing, deletion of an entire coding sequence in the SLC2A1 gene was detected. Early introduction of a modified Atkins diet maintained a seizure-free period without antiseizure medications and normal cognitive development in the follow-up period. Our report summarizes the clinical features of GLUT1 syndromes and discusses the importance of early identification and molecular confirmation of GLUT1DS as a treatable metabolic disorder.

Epilepsy is the only chronic neurological disease that can be fully controlled by drugs or introduced into permanent remission. Nonetheless, pharmacological treatment of epilepsy is often accompanied by a range of diagnostic and therapeutic challenges. Due to a wide variety of available antiepileptics, the selection of the first antiepileptic, as well as the combination of polytherapy, is extremely large. The choice of the ideal antiepileptic is individual and depends on a variety of factors based on the pharmacological properties of the drug and the disctinctiveness of every patient (type of seizure, age,sex, profession, comorbidity, co medication, etc.) with specific regulatory limitations (availability,price, indications of regulatory bodies). The basics of rational pharmacotherapy include slow titration of antiepileptics in monotherapy to satisfactory effect, slow drug replacement in the event of failure of the first antiepileptic, combination of two antiepileptics of different mechanism of action or selection of synergistic combinations if dual therapy is necessary, avoiding three or multiple polyterapies whenever possible, and slow reduction of antiepileptic drug after achieving a long-lasting remission.Current ILAE guidelines for epilepsy treatment imply basic and comprehensive recommendations that are acceptable worldwide, including countries with the lowest economic standard. Therefore, these guidelines do not reflect the ideal treatment options, especially in developed countries, and are actually the basis for making national guidelines. The Croatian guidelines for pharmacological treatment of epilepsy are the product of the co-operation of all relevant professional societies and reference centers in the Republic of Croatia, headed by the Croatian League Against Epilepsy and the Croatian Neurological Society. These guidelines reflect the current socioeconomic specificities in our country,the latest knowledge of pharmacological profiles, and efficacy of certain antiepileptics as well as expert opinions.

Neuronska ceroidna lipofuscinoza-tip 2 (CLN2) najčešća je dječja progresivna neurodegenerativna bolest. Glavna obilježja bolestiCLN2 su usporen razvoj govora koji prethodi epileptičkim napadajima, motoričkim poremećajima, progresivnom propadanju vida ikognitivnih funkcija te smrti u dobi rane adolescencije. Prosječno vrijeme od pojave prvih simptoma do postavljenja dijagnoze iznosi oko dvije godine. Kašnjenje u postavljanju dijagnoze odgađa terapijski pristup koji uključuje multidisciplinsku zdravstvenu skrb iučinkovitu primjenu enzimske nadomjesne terapije cerliponazom alfa (Brineura®). Dostupnost liječenja enzimskom nadomjesnomterapijom cerliponazom alfa svrstava CLN2 u skupinu lječivih neurodegenerativnih bolesti. U svake lječive i/ili potencijalno izlječiveprogresivne neurodegenrativne bolesti pravodobna dijagnoza je ključna, jer omogućuje liječenje u ranom stadiju bolesti. Prepoznavanje ranih simptoma od bitnog je značenja za usmjeravanje kliničara na odgovarajuće pretrage ili traženje dodatnog stručnogmišljenja. U sve djece s prvim neprovociranim epileptičkim napadajem u dobi između druge i četvrte godine, a izričito u one djecekoja dodatno u anamnezi imaju podatak o usporenom razvoju govora i/ili motoričke smetnje prvi korak u cilju rane i pravodobnedijagnoze je mjerenje aktivnosti enzima TTP1 putem suhe kapi krvi na fi ltarskom papiru, koji je dostupan u našoj sredini. Dodatnagenska analiza i nalaz patogenih mutacija u genu CLN2 potvrđuje dijagnozu. Podizanje svjesnosti o bolesti CLN2 kao lječivoj i potencijalno izlječivoj, te poštivanje predloženih smjernica u kliničkom pristupu zasigurno će pridonijeti tome da rana dijagnoza bolestiCLN2 postane naša stvarnost, a sve manje bude mit. Pravovremena i ispravna dijagnoza prvi je korak u poboljšanju cjelokupne skrbiza djecu oboljelu od bolesti CLN2 i njihove roditelje.

Hypothalamic orexin neurons project to many brain areas, including hindbrain structures such as the nucleus of the solitary tract (NTS) and area postrema (AP), where orexin 1 receptors (OX1Rs) are expressed. Hindbrain administration of orexin-A increases feeding and meal size, and blockade of hindbrain OX1Rs with the selective antagonist SB334867 has the opposite effect. Here we asked whether hindbrain OX1R stimulation or blockade alter rats’ sensitivity to gastrointestinal satiety signals. Rats received 4th intracerebroventricular (icv) injections of vehicle or orexin-A, at a dose with no effect on its own, prior to an intragastric (IG) infusion of saline or a satiating volume of Ensure. IG Ensure suppressed subsequent chow intake, but orexin-A pretreatment significantly attenuated this IG nutrient-induced satiety at 2 h into the dark phase. In a second experiment, rats received NTS injections of vehicle or orexin-A before intraperitoneal (IP) injection of vehicle or the satiation hormone cholecystokinin (CCK). NTS orexin-A pretreatment completely blocked the intake-suppressive effect of CCK on dark-phase chow intake. Finally, we investigated the role of endogenous hindbrain OX1R activation by pretreating rats with 4th-icv injection of vehicle or SB334867 followed by IG infusion of saline or Ensure just before a chocolate Ensure licking test session. IG nutrient infusion suppressed Ensure intake, and blockade of hindbrain OX1Rs significantly prolonged that intake-suppressive effect. We conclude that hindbrain OX1Rs are a mechanism though which hypothalamic orexin neurons can reduce animals’ sensitivity to gastrointestinal nutrient load, allowing them to consume more food.