Microencapsulation of parathyroid cells via electric field and non-surgical transplantation approach.

PURPOSE: Hypoparathyroidism is a rare disease with low PTH, mostly seen as a consequence of neck surgery. Current management is the prescription of calcium and vitamin D, but the definitive treatment is parathyroid allotransplantation, which frequently triggers an immune response, thus cannot achieve the expected success. To overcome this problem, encapsulation of allogeneic cells is the most promising method. By optimizing the standard alginate cell encapsulation technique with parathyroid cells under high-voltage application, the authors reduced the size of parathyroid-encapsulated beads and evaluated these samples in vitro and in vivo. METHODS: Parathyroid cells were isolated, and standard-sized alginate macrobeads were prepared without any electrical field application, while microbeads in smaller sizes (< 500 µm), by the application of 13 kV. Bead morphologies, cell viability, and PTH secretion were evaluated in vitro for four weeks. For the in vivo part, beads were transplanted into Sprague-Dawley rats, and after retrieval, immunohistochemistry and PTH release were evaluated in addition to the assessment of cytokine/chemokine levels. RESULTS: The viability of parathyroid cells in micro- and macrobeads did not differ significantly. However, the amount of in vitro PTH secretion from microencapsulated cells was significantly lower than that from macroencapsulated cells, although it increased throughout the incubation period. Immunohistochemistry of PTH staining in both of the encapsulated cells identified as positive after retrieval. CONCLUSION: Contrary to the literature, a minimal in vivo immune response was developed for alginate-encapsulated parathyroid cells, regardless of bead size. Our findings suggest that injectable, micro-sized beads obtained using high-voltage may be a promising method for a non-surgical transplantation approach.

Paracrine effects of living human bone particles on the osteogenic differentiation of mesenchymal stem cells.

Bone autografting remains the clinical model of choice for resolving problematic fractures. The precise mechanisms through which the autograft promotes bone healing are unknown. The present study examined the hypothesis that cells within the autograft secrete osteogenic factors promoting the differentiation of mesenchymal stem cells (MSCs) into osteoblasts. Particles of human bone ("chips") were recovered at the time of joint replacement surgery and placed in culture. Then, conditioned media were added to cultures of human, adipose-derived MSCs under both basal and osteogenic conditions. Contrary to expectation, medium conditioned by bone chips reduced the expression of alkaline phosphatase and strongly inhibited mineral deposition by MSCs cultured in osteogenic medium. Real time PCR revealed the inhibition of collagen type I alpha 1 chain (Col1A1) and osteopontin (OPN) expression. These data indicated that the factors secreted by bone chips inhibited the osteogenic differentiation of MSCs. However, in late cultures, bone morphogenetic protein-2 (BMP-2) expression was stimulated, suggesting the possibility of a delayed, secondary osteogenic effect.

The effect of accelerated mineral trioxide aggregate on odontoblastic differentiation in dental pulp stem cell niches.

AIM: To investigate the effect of accelerated-set mineral trioxide aggregate (MTA) on the proliferation and odontoblastic differentiation of human dental pulp cell niches (DPSC). METHODOLOGY: ProRoot White MTA (WMTA; Dentsply Tulsa Dental, Johnson City, TN, USA) was mixed with various additives, which included distilled water, 2.5% disodium hydrogen phosphate (Na2 HPO4 ; Merck, Darmstadt, Germany) and 5% calcium chloride (CaCl2 ; Merck). DPSC niches extracted from third molars were cultured directly on MTA in the culture medium. Cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4- sulphophenyl)-2H-tetrazolium (MTS) assay. Cell growth and expression of odontoblastic differentiation markers (dentine sialophosphoprotein (DSPP) and collagen type 1 (COL1)) were determined using Real-Time Polymerase Chain Reaction analysis. Osteo-/odontogenic differentiation of DPSC niches was evaluated by measurement of alkaline phosphatase activity (ALP). Calcium deposition was assessed using von Kossa staining. The results were analysed statistically using Mann-Whitney tests and Kruskal-Wallis tests. RESULTS: MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 exhibited optimal cell viability (P < 0.05) compared to MTA mixed with distilled water. MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 significantly increased ALP activity (P < 0.05), significantly promoted mineralization nodule formation (P < 0.05) and significantly enhanced the mRNA expression level of the osteogenic/odontogenic markers (P < 0.05; DSPP and COL1) compared with MTA mixed with distilled water. CONCLUSIONS: MTA mixed with 5% CaCl2 and 2.5% Na2 HPO4 was biocompatible with dental pulp stem cell niches. Accelerated-set MTA promoted better differentiation in DPSC niches than conventional MTA. The accelerators could provide an alternative to MTA mixed with distilled water.

Chondrogenic potential of macroporous biodegradable cryogels based on synthetic poly(α-amino acids).

In this study, the potential of highly porous hydrogels based on biodegradable synthetic poly(α-amino acids) to support proliferation and chondrogenesis of human dental pulp stem cells (hDPSCs) was investigated. Covalently crosslinked gels with permanent pores were formed under cryogenic conditions by free-radical copolymerization of poly[N5-(2-hydroxyethyl)-l-glutamine-stat-N5-(2-methacryloyl-oxy-ethyl)-l-glutamine] (PHEG-MA) with 2-hydroxyethyl methacrylate (HEMA) and N-propargyl methacrylamide (PrMAAm) as minor co-monomers. PrMAAm provided alkyne groups for modifying the gels with cell-supporting moieties (RGDS peptides) by the azide-alkyne "click"-reaction. Two types of gels with different compressive moduli were prepared. Each type was modified with two different concentrations of RGDS peptide. X-ray computed nanotomography (nanoCT) was used to visualize and analyze the 3D-structure of the cryogels. It was shown that modifying the PHEG-MA cryogels within the range of RGDS concentrations examined here had a positive effect on the proliferation of hDPSCs. Immunofluorescence staining for collagen type 2 and aggrecan proved that there was differentiation of hDPSCs into chondrocytes.

Cartilage tissue engineering on macroporous scaffolds using human tooth germ stem cells.

The main objective was to study cartilage regeneration through differentiation of human tooth germ stem cells (HTGSCs) into chondrocytes on different three-dimensional (3D) scaffolds (PCL, PLLA and PCL-PLLA). Scaffold topographies were studied by scanning electron microscopy and it was found that the scaffolds had interconnected macroporous structures. HTGSCs were isolated from impacted third molar tooth germs of young adult patients and grown for 3 weeks on the scaffolds in chondrogenic differentiation medium. Cell proliferation on the scaffolds was determined by MTS assay and it was observed that all scaffolds supported cell proliferation. Immunostaining was carried out for morphological and differentiation analyses. Immunohistochemical analyses revealed that the cells attached onto the scaffolds and deposited cartilage-specific extracellular matrix (ECM). Real-time PCR was performed to determine the expression levels of cartilage-specific genes. After 21 days of incubation in cartilage differentiation medium, expression of collagen type II increased only in the cells seeded onto PCL-PLLA blend scaffolds. Similarly, aggrecan expression was the highest on PCL-PLLA scaffolds after 3 weeks. These results suggest that all the scaffolds, and especially PCL-PLLA, were suitable for chondrogenic differentiation of HTGSCs. Copyright © 2015 John Wiley & Sons, Ltd.

The ovarian cancers in geriatric population: the validity of inflammatory markers, malignancy risk indices 1, 2, 3, 4, and CA-125 levels in malignancy discrimination of adnexal masses.

PURPOSE: To investigate the predictive value of the Risk of Malignancy Index (RMI), CA-125, and inflammatory markers in discriminating ovarian cancers (OCs). MATERIALS AND METHODS: The postmenopausal (PM) women (n= 139) with adnexal masses who un- derwent surgery were included. The predictive value of CA-125, RMI (1, 2,3, and 4) and inflammatory markers [neutrophil lymphocyte ratio (NLR), platelet lymphocyte ratio (PLR)] were calculated in geriatric (G) and non-geriatric women. RESULTS: OCs had significantly increased NLR and PLR. RMI models were highly reliable in PM (Kappa: 0.642-0.715; AUC: 0.907-0.934). CA-125 measurement alone had good accuracy and moderate reliability in PM (kappa: 0.507-0.587), excellent accuracy and moderate reliability in G, NLR, and PLR predicting OCs, showed fair agreement in the PM, while PLR had a moderate agreement with G. CONCLUSION: RMI algorithms were the best models for malignancy prediction. However, the rise of PLR and CA-125 levels in a G population may be used as refer- ring adnexal masses to gynecologic oncologists.

Perinatal and neonatal outcomes of maternal heart diseases.

AIMS: To explore the perinatal and neonatal outcomes of patients with heart diseases. Materials Methods: Retrospective case control analysis was carried out among 10,527 deliveries, 188 pregnancies complicated by cardiovascular disease (CVD) compared with pregnancies without CVD for obstetric outcomes from January 2000 to December 2012. The effect of cardiac functional classification (NHYA) on maternal and neonatal complications was explored. RESULTS: The incidence of CVD in pregnancy was 1.78%. About 80.3% had rheumatic heart disease (RHD). Maternal and neonatal mortality rate was 1.06% and 2.13 %, respectively. The obstetric outcomes of women in NHYA class I/II were similar to normal group. Vaginal delivery was the preferred way of birth unless deterioration of cardiac functions as in the cases of NHYA class III/IV. NHYA class III/IV had significantly decreased birth weight, premature birth, and increased maternal-neonatal mortality (p < 0.05). CONCLUSION: RHD is still prevalent. The cardiac functional capacity predicts maternal and neonatal outcomes.

Trace element levels in children with atopic dermatitis.

BACKGROUND: Trace elements are micronutrients that are present in small amounts in the body and are essential for normal functioning of the immune and antioxidant systems. Inflammation and oxidative stress are major pathogenic mechanisms in the development of atopic dermatitis (AD). The role of micronutrients in AD has been investigated in a limited number of studies, although the results are contradictory. OBJECTIVES: In this study, we examined the levels of iron, copper, and magnesium in serum and the level of zinc in erythrocytes in children with AD. We compared our findings with those of a healthy control group. METHOD: The study population comprised 92 AD patients and 70 controls. We performed a complete blood count and measured levels of iron, copper, and magnesium in serum and levels of zinc in erythrocytes. RESULTS: We found that serum magnesium and erythrocyte zinc levels were lower in children with AD than in the control group; levels of copper and iron did not differ between the groups. The levels of micronutrients studied were not correlated with disease severity. CONCLUSION: Evaluation of zinc and magnesium levels in children with AD could prove useful. The role of micronutrients in the pathogenesis and course of AD warrants further study.

Specific immunotherapy improves asthma related quality of life in childhood

Background: Asthma is the most common chronic illness in childhood and really affects the everyday life of patients who suffer from it. Since asthma is a common disease, there is a great endeavour to achieve the most appropriate treatment option. Despite inhaled corticosteroids and leukotrien receptor antagonists both being routinely used in asthma treatment, specific immunotherapy is still questioned. There are numerous aspects affecting asthma-related quality of life, such as age; seasons; disease control and severity etc, which are well studied -apart from the type of treatment. With this study we aimed to stress the influence of asthma treatment on quality of life. Methods: A total of 102 children, aged 6—18 years, were assigned to classic asthma therapy (n=50) and specific immunotherapy (n=52). The quality of life is assessed using the Standardized Pediatric Asthma Quality of Life Questionnaire (PAQLQ) interviewer-administered Turkish version. Pulmonary function testing was performed on the same day, after the questionnaire was completed. Results: The PAQLQ total scores were significantly higher in the specific immunotherapy group (p<0.001). Apart from emotional function domain scores; symptoms domain and activity limitation domain scores were higher in the specific immunotherapy group. Emotional function domain scores were similar in the two groups (p>0.05). There were no statistically significant differences in pulmonary function testing results between the two groups (p>0.05). There was a linear correlation between FEV1%, FVC level and total and domain scores of PAQLQ with Spearman Correlation tests (AU)

Specific immunotherapy improves asthma related quality of life in childhood.

BACKGROUND: Asthma is the most common chronic illness in childhood and really affects the everyday life of patients who suffer from it. Since asthma is a common disease, there is a great endeavour to achieve the most appropriate treatment option. Despite inhaled corticosteroids and leukotrien receptor antagonists both being routinely used in asthma treatment, specific immunotherapy is still questioned. There are numerous aspects affecting asthma-related quality of life, such as age; seasons; disease control and severity etc, which are well studied -apart from the type of treatment. With this study we aimed to stress the influence of asthma treatment on quality of life. METHODS: A total of 102 children, aged 6-18 years, were assigned to classic asthma therapy (n=50) and specific immunotherapy (n=52). The quality of life is assessed using the Standardized Pediatric Asthma Quality of Life Questionnaire (PAQLQ) interviewer-administered Turkish version. Pulmonary function testing was performed on the same day, after the questionnaire was completed. RESULTS: The PAQLQ total scores were significantly higher in the specific immunotherapy group (p<0.001). Apart from emotional function domain scores; symptoms domain and activity limitation domain scores were higher in the specific immunotherapy group. Emotional function domain scores were similar in the two groups (p>0.05). There were no statistically significant differences in pulmonary function testing results between the two groups (p>0.05). There was a linear correlation between FEV1%, FVC level and total and domain scores of PAQLQ with Spearman Correlation tests.

Design of a 3D aligned myocardial tissue construct from biodegradable polyesters.

The heart does not regenerate new functional tissue when myocardium dies following coronary artery occlusion, or if it is defective. Ventricular restoration involves excising the infarct and replacing it with a cardiac patch to restore the heart to a more healthy condition. The goal of this study was to design and develop a clinically applicable myocardial patch to replace myocardial infarcts and improve long-term heart function. A basic design composed of 3D microfibrous mats that house mesenchymal stem cells (MSCs) was developed from human umbilical cord matrix (Wharton's Jelly) cells aligned in parallel to each other mimicking the native myocardium. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), poly(L-D,L-lactic acid) (P(L-D,L)LA) and poly(glycerol sebacate) (PGS) were blended and electrospun into aligned fiber mats with fiber diameter ranging between 1.10 and 1.25 microm. The micron-sized parallel fibers of the polymer blend were effective in cell alignment and cells have penetrated deep within the mat through the fiber interstices, occupying the whole structure; 8-9 cell layers were obtained. Biodegradable macroporous tubings were introduced to serve as nutrient delivery route. It was possible to create a thick myocardial patch with structure similar to the native tissue and with a capability to grow.

Isolation and characterization of stem cells derived from human third molar tooth germs of young adults: implications in neo-vascularization, osteo-, adipo- and neurogenesis.

A number of studies have reported in the last decade that human tooth germs contain multipotent cells that give rise to dental and peri-odontal structures. The dental pulp, third molars in particular, have been shown to be a significant stem cell source. In this study, we isolated and characterized human tooth germ stem cells (hTGSCs) from third molars and assessed the expression of developmentally important transcription factors, such as oct4, sox2, klf4, nanog and c-myc, to determine their pluri-potency. Flow-cytometry analysis revealed that hTGSCs were positive for CD73, CD90, CD105 and CD166, but negative for CD34, CD45 and CD133, suggesting that these cells are mesenchymal-like stem cells. Under specific culture conditions, hTGSCs differentiated into osteogenic, adipogenic and neurogenic cells, as well as formed tube-like structures in Matrigel assay. hTGSCs showed significant levels of expression of sox2 and c-myc messenger RNA (mRNA), and a very high level of expression of klf4 mRNA when compared with human embryonic stem cells. This study reports for the first time that hTGSCs express developmentally important transcription factors that could render hTGSCs an attractive candidate for future somatic cell re-programming studies to differentiate germs into various tissue types, such as neurons and vascular structures. In addition, these multipotential hTGSCs could be important stem cell sources for autologous transplantation.

A Feingold syndrome case with previously undescribed features and a new mutation.

Feingold syndrome (FS) is a dominantly inherited combination of microcephaly with or without learning disabilities, hand and foot abnormalities, short palpebral fissures and esophageal/duodenal atresia. The syndrome has autosomal dominant inheritance with full penetrance, and variable expressivity. Digital anomalies are almost always present. The gene for FS is localized to a 2.2 cM region in 2p23-p24. We report on the first Turkish family with Feingold syndrome. The propositus is a male infant with microcephaly, frontal balding, brachymesophalangy of the second and fifth fingers, bilateral syndactyly of toes 2-3, facial anomalies, choanal atresia and focal epilepsy. His father has microcephaly, and more severe hands and feet abnormalities. One of his brothers died because of eosofageal atresia. Clinical presentation of the family was suggestive of Feingold syndrome, and genetic testing of the MYCN gene confirmed the diagnosis. The missense mutation we report here has not been described previously. FS is an autosomal dominant condition, and therefore, the diagnosis has important implications for genetic counseling.

Ultrasound-guided botulinum toxin type A injection to the iliopsoas muscle in the management of children with cerebral palsy.

OBJECTIVE: To determine the effect of botulinum toxin type A (BTX-A) on spasticity and functional development in children with cerebral palsy (CP) in conjunction with a physiotherapy program. METHOD: In this prospective study, 18 CP patients were evaluated. Multilevel BTX-A injection was applied to children at a dose of 15 U/kg. Children were assessed before and at the 5th and 12th week post-injection using Thomas test, Duncan-Ely test, passive range of motion (pROM) measurement, Distance Between Knee (DBK), Selective Motor Control (SMC) scale, modified Ashworth Scale (MAS) and modified Physician Rating Scale (mPRS). To assess functional improvement, Gross Motor Function Measure (GMFM) and Functional Independence Measure for Children (WeeFIM) were used before and at the 12th week post-injection. RESULTS: At 5th week post-injection, a statistically significant decrease was determined in spasticity (p < 0.01). Improvement was observed in mPRS and pROM, but not in SMC. At the 12th week post-injection, GMFM (p< 0.001) and WeeFIM improved significantly (p< 0.001). The improvement in pROM and mPRS (p< 0.01) lasted until the 12th week post-injection, but the improvement in MAS (p > 0.05) and in the Tardieu test of hip adductors (p > 0.05) did not last after the 5th week. CONCLUSION: BTX-A injection enhances functional and motor abilities in the development process.

The subclinical incidence of CTS in pregnancy: Assessment of median nerve impairment in asymptomatic pregnant women.

PURPOSE: The true incidence of pregnancy related carpal tunnel syndrome (PRCTS) is unknown. Most of the diagnoses of PRCTS are made based only on clinical symptoms. Here, we report a prospective controlled clinical trial assessing the electrophysiological changes in pregnant women to provide objective measure of the median nerve function. METHODS: Pregnant women in the third trimester (n=69) and age-matched non-pregnant women (n=40) asymptomatic for CTS were included in the study. Nerve conduction studies of the median and ulnar nerves across the carpal tunnel were bilaterally performed with the standard techniques. RESULTS: All the median sensory nerve conduction studies (amplitude, latency and velocity) performed from the ring finger and palmar region to wrist showed significant prolongation of median nerve conduction in the pregnant women compared with the control group (*p

Giant axonal neuropathy: clinical and genetic study in six cases.

BACKGROUND: Giant axonal neuropathy (GAN) is a severe recessive disorder characterised by variable combination of progressive sensory motor neuropathy, central nervous system (CNS) involvement, and "frizzly" hair. The disease is caused by GAN gene mutations on chromosome 16q24.1. AIMS: To search for GAN gene mutations in Turkish patients with GAN and characterise the phenotype associated with them. METHODS: Linkage and mutation analyses were performed in six affected patients from three consanguineous families. These patients were also investigated by cranial magnetic resonance imaging (MRI) and electroencephalography (EEG). Electromyography (EMG) was performed in heterozygous carriers from family 1 and family 3. RESULTS: Linkage to 16q24.1 was confirmed by haplotype analysis. GAN mutations were identified in all families. Family 1 had the R293X mutation, previously reported in another Turkish family. Families 2 and 3, originating from close geographical areas, shared a novel mutation, 1502+1G>T, at the donor splice site of exon 9. All patients displayed a common phenotype, including peripheral neuropathy, cerebellar ataxia, and frizzly hair. Cranial MRI showed diffuse white matter abnormalities in two patients from family 1 and the patient from family 3, and minimal white matter involvement in the patient from family 2. EMG of a heterozygous R293X mutation carrier showed signs of mild axonal neuropathy, whereas a 1502+1G>T mutation carrier had normal EMG. EEG abnormalities were found in three patients. CONCLUSION: These findings highlight the association of CNS involvement, in particular white matter abnormalities, with peripheral neuropathy in GAN. The phenotypical consequences of both mutations (when homozygous) were similar.

Bone tissue engineering on patterned collagen films: an in vitro study.

This study aimed at guiding osteoblast cells from rat bone marrow on chemically modified and patterned collagen films to study the influence of patterns on cell guidance. The films were stabilized using different treatment methods including crosslinking with carbodiimide (EDC) and glutaraldehyde, dehydrothermal treatment (DHT), and deposition of calcium phosphate on the collagen membrane. Mesenchymal osteoprogenitor cells were differentiated into osteoblasts and cultured for 7 and 14 days on micropatterned (groove width: 27 microm, groove depth: 12 microm, ridge width: 2 microm) and macropatterned (groove width: 250 microm, groove depth: 250 microm, ridge width: 100 microm) collagen films to study the influence of pattern dimensions on osteoblast alignment and orientation. Fibrinogen was added to the patterned surfaces as a chemical cue to induce osteoblast adhesion. Cell proliferation on collagen films was determined using MTS assay. Deposition of calcium phosphate on the surface of the film increased surface hydrophilicity and roughness and allowed a good cell proliferation. Combined DHT and EDC treatment provided an intermediate wettability, and also promoted cell proliferation. Glutaraldehyde crosslinking was found to lead to the lowest cell proliferation but fibrinogen adsorption on glutaraldehyde treated film surfaces increased the cell proliferation significantly. Macropatterns were first tested for alignment and only microscopy images were enough to see that there is no specific alignment. As a result of this, micropatterned samples with the topography that affect cell alignment and guidance were used. Osteoblast phenotype expression (ALP activity) was observed to be highest in calcium phosphate deposited samples, emphasizing the effect of mineralization on osteoblast differentiation. In general ALP activity per cell was found to decrease from day 7 to day 14 of incubation. SEM and fluorescence microscopy revealed good osteoblast alignment and orientation along the axis of the patterns when micropatterned films were used. This study shows that it is possible to prepare cell carriers suitable for tissue engineering through choice of appropriate surface topography and surface chemistry. Presence of chemical cues and micropatterns on the surface enhance cell orientation and bone formation.

Bone generation on PHBV matrices: an in vitro study.

Bone formation was investigated in vitro by culturing rat marrow stromal osteoblasts in biodegradable, macroporous poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) matrices over a period of 60 days. Foams were prepared after solvent evaporation and solute leaching. PHBV solutions with different concentrations were prepared in chloroform: dichloromethane (1:2, v/v). In order to create a matrix with high porosity and uniform pore sizes, sieved sucrose crystals (300-500 microm) were used. PHBV foams were treated with rf-oxygen plasma (100 W 10 min) to modify their surface chemistry and hydrophilicity with the aim of increasing the reattachment of osteoblasts. Osteoblasts were isolated from rat bone marrow and seeded onto PHBV foams. The cell density on and in the foams was determined with MTS assay. MTS results showed that osteoblasts proliferated on PHBV. Twenty-one days after seeding of incubation, growth of osteoblasts on matrices and initiation of mineralization were observed by confocal laser scanning microscopy. Increasing ALP and osteocalcin secretion during 60 days confirmed the osteoblastic phenotype of the derived stromal cells. SEM, histological evaluations and confocal laser scanning microscopy showed that osteoblasts could grow inside the matrices and lead to mineralization. Cells exhibited spindle-like morphology and had a diameter of 10-30 microm. Based on these, it could confidently be stated that PHBV seems to be a promising polymeric matrix material for bone tissue engineering.

Acute disseminated encephalomyelitis in children: outcome and prognosis.

BACKGROUND: Acute disseminated encephalomyelitis (ADEM) is the most common demyelinating disorder of childhood. Its clinical features, prognosis and treatment vary in different reports. OBJECTIVES: To examine a series of children with ADEM for clinical findings, course, recurrences, and possible variables affecting outcome. METHODS: Multicentric data collected from 7 tertiary referral centers were registered and evaluated in a central database in 1990 - 2001 for clinical, laboratory, and MRI features. Course and prognosis were assessed in patients with at least 12 months' follow-up. RESULTS: Forty-six patients were evaluated. Median age at onset was 8 years, M/F ratio, 1.7/1. Most common symptoms and signs pertained to the motor system and consciousness. Of 39 children with 12 months' follow-up, 71 % recovered completely. Thirteen (33 %) children had relapses. Patients who had more than one relapse (n = 4) presented with new symptoms at each attack. Treatment with high-dose methylprednisolone was associated with complete recovery, and tapering over more than 3 weeks, with a lower rate of relapses. MRI lesions could persist even in asymptomatic patients; in particular, periventricular lesions tended to disappear later than others. CONCLUSIONS: Complete clinical recovery is common and serious complications are rare in childhood ADEM, but the rate of relapses is considerable. Clinical picture at first relapse may help to identify patients likely to experience multiple relapses. The timing and duration of steroid treatment affects outcome.