Diatom extraction: A new technique with heated H2O2. A technical note.

The best method of diatom identification in animal and human tissues is still an important discussion topic, in terms of effectiveness and reliability. In this technical note, authors propose a new method of extraction of diatoms using heated hydrogen peroxide from animal and human tissue samples. This method has been compared with the traditional method of digestion with acids. The results of the comparison show that heated hydrogen peroxide extraction is more efficient in terms of reduction of sediment, extraction of the material and preservation of diatoms proving to be a viable alternative to conventional approaches with acids in terms of costs and operator safety.

Interferon-related transcriptome alterations in the cerebrospinal fluid cells of Aicardi-Goutières patients.

Aicardi-Goutières syndrome (AGS) is a rare interferon (IFN)-related encephalopathy with onset during the first year of life. AGS, is clinically characterized by progressive microcephaly, bilateral basal ganglia calcification, cerebral atrophy, cerebrospinal fluid (CSF), lymphocytosis, delayed development of psychomotor abilities with pyramidal-extrapyramidal syndrome and mimics congenital viral infections. Microarray analysis examining the expression of 18 880 human genes has been applied to the CSF lymphocytes of 20 AGS cases (age 4.5 +/- 4.4 years, mean +/- standard deviation) characterized by high IFN-alpha levels in CSF and 20 matched controls (age 4.4 +/- 4.3 years, mean +/- standard deviation). Gene-expression data reveal significant differences between AGS cases and controls for all controls and 18 AGS cases. The two AGS cases unclassified as compared with controls were both older than 7 years. AGS cases presented upregulation of genes involved in IFN-dependent pathways and lymphocyte functions, paralleled by the downregulation of genes encoding for angiopoietic activities. The cystatin F and DNAJ genes, having a negative feedback on IFN pathways, underwent a progressive age-related increase in their expression. These gene-expression signature parallels a progressive attenuation of clinical symptoms with age. Obtained results provide evidence that exposure to IFN-alpha is harmful for developing brain.

The Rorschach test and Gilles de la Tourette's syndrome: a pilot case-control study.

BACKGROUND: To date only three studies, all lacking a control group, have investigated the personality style underlying tic disorders or Tourette's syndrome (TS), using the projective Rorschach test. Despite the recent controversy about its proper use in clinical practice and research, the Rorschach test may be useful to explore personality organization in TS. The research hypothesis under investigation in this exploratory study was that young TS patients exhibit a peculiar personality organization, with significantly increased indicators of poor emotional control, aggression, anxiety and depression, on the Rorschach test, compared to healthy controls. METHODS: We analyed the Rorschach protocols of 17 newly diagnosed paediatric TS patients and 17 age- and sex-matched healthy controls. RESULTS: Compared with the controls young TS patients recorded significantly lower mean percentages of F and F+ responses and a significantly higher number of FM and FC responses, thus showing reduced control of emotions and difficulty in integrating aggressivity. Furthermore, associations emerged between comorbidity with ADHD and increased emotional instability and between severity of tics and of OCD and a pattern of emotional hyper-control and application of rigid defensive mechanisms. CONCLUSIONS: These results seem to confirm the existence of a peculiar personality style in young TS patients and may have important implications, both pathogenetic and therapeutic.

Dopaminergic receptor D5 mRNA expression is increased in circulating lymphocytes of Tourette syndrome patients.

Tourette syndrome (TS) is a neuropsychiatric disorder in which dopaminergic dysfunction and immune system abnormalities seem to coexist. Using real-time PCR, we determined mRNA expression of dopamine receptors (DRs) D1-5 in peripheral blood lymphocytes (PBLs) from 15 TS patients and 15 sex- and age-matched healthy controls (HCs). DRD5 mRNA levels in cells from TS were higher than in cells from HCs. In TS patients with obsessive-compulsive disorder, DRD5 mRNA levels in PBLs showed a highly positive correlation with the severity of compulsive symptoms. DRD5 mRNA upregulation in PBLs from TS patients may represent a peripheral marker of dopaminergic dysfunction and supports the involvement of the immune system in TS.

The development of visual object recognition in school-age children.

This study documents the age-dependent development of visual object recognition abilities in 115 children aged 6 to 11 years, using a battery of neuropsychological tests based on Marrs model (Efron test, Warringtons Figure-Ground Test, Street Completion Test, Poppelreuter-Ghent Test, a selection of stimuli from the Birmingham Object Recognition Battery, a series of color photographs of objects presented from unusual perspectives or illuminated in unusual ways). The results suggest a maturation of complex visual perceptual abilities, possibly related to the development of the cerebral processes involved in object recognition, and could be the starting point for future investigations of these skills in impaired populations.

Neuropsychological deficits and neural dysfunction in familial dyslexia.

We report the neuropsychological profile and the pattern of brain activity during reading tasks in a sample of familial dyslexics. We studied our subjects with an in-depth neuropsychological assessment and with functional neuroimaging (fMRI) during word and pseudoword reading and false font string observations (baseline condition). The neuropsychological assessment revealed that familial dyslexia, in both persistent and compensated forms, is often associated with deficits in verbal short-term memory, phonological awareness and automatization abilities. The functional results showed a lack of activation in the posterior areas of the reading network. This study, together with the previously published VBM study (Brambati, S.M., Termine, C., Ruffino, M., Stella, G., Fazio, F., Cappa, S.F. and Perani, D., Regional reductions of gray matter volume in familial dyslexia, Neurology, 63 (2004) 742-5), provides a multiple modality evaluation of familial dyslexia. The neuropsychological assessment showed cognitive deficits associated with dyslexia that persist also in subjects with compensated reading deficit. Both the anatomical and the functional study point out a deficit in the posterior areas of the reading network.

Mutations in genes encoding ribonuclease H2 subunits cause Aicardi-Goutières syndrome and mimic congenital viral brain infection.

Aicardi-Goutières syndrome (AGS) is an autosomal recessive neurological disorder, the clinical and immunological features of which parallel those of congenital viral infection. Here we define the composition of the human ribonuclease H2 enzyme complex and show that AGS can result from mutations in the genes encoding any one of its three subunits. Our findings demonstrate a role for ribonuclease H in human neurological disease and suggest an unanticipated relationship between ribonuclease H2 and the antiviral immune response that warrants further investigation.

Relationship between migraine and epilepsy in pediatric age.

OBJECTIVE: Many studies have supported the hypothesis of alteration of cortical hyperexcitability as a possible pathological mechanism underlying the onset of migraine and epileptic attacks. Different biochemical pathways involving cellular structures may increase or decrease the excitability of neuronal membranes. The aim of this study was to identify a possible link between migraine and epilepsy from a clinical and neurophysiologic point of view. METHODS: One-hundred thirty-seven children and adolescents consecutively diagnosed for tension-type headache and idiopathic migraine with and without aura were studied. Anamnestic, clinical, and instrumental data were collected by a neurological examination, a specific questionnaire, and awake electroencephalogram (EEG) registrations. EEG features of nonheadache and nonseizures control group were compared. RESULTS: Fourteen cases (10.2%) had a positive history for seizures with fever, isolated seizures, or epilepsy. Distribution differed according to headache diagnosis; migraine with aura (MWA) was largely prevalent. Specific electroencephalographic abnormalities were present in 11.7% of the sample, with a significant different distribution across the groups of children with headache and the control group: specific interictal abnormalities were found in 10 of 23 (43.5%) children with MWA. Two factors, seizures and specific interictal electroencephalographic abnormalities, showed a different distribution in patients with MWA compared to other classes of headache and control group (P < or = .01). CONCLUSION: The present study supports the hypothesis of a possible clinical continuum between some types of MWA and epileptic syndromes as entities due to altered neuronal excitability with similar genetic substrates.

Bilateral putaminal necrosis associated with the mitochondrial DNA A8344G myoclonus epilepsy with ragged red fibers (MERRF) mutation: an infantile case.

Myoclonus epilepsy with ragged red fibers (MERRF) is one of the major mitochondrial encephalomyopathies. Its main clinical features are myoclonus epilepsy, ataxia, and myopathy with ragged red fibers. Whereas there is a close correlation between MERRF syndrome and the A8344G mutation of mitochondrial DNA, the reverse is not true. In fact, this mutation is also responsible for various other syndromes, such as Leigh syndrome, spinocerebellar degeneration, atypical Charcot-Marie-Tooth disease, and multiple truncal lipomas. We describe a child with the A8344G mutation of mitochondrial DNA and an unusual clinical, neuroradiologic, and biochemical phenotype, characterized by early-onset, nonprogressive cerebellar ataxia, and subclinical myoclonias in association with bilateral putaminal necrosis on magnetic resonance imaging and a reduction in complex V activity. Our case confirms the existence of a relationship between alteration in adenosine triphosphatase activity and basal ganglia involvement. We recommend that the possibility of a mitochondrial pathology should always be taken into consideration in the presence of bilateral symmetric lesions of the basal ganglia, even when the typical clinical picture is lacking. (J Child Neurol 2006;21:79-82).

A pilot study of a ketogenic diet in patients with Lafora body disease.

PURPOSE: Lafora body disease (LBD) is severe and rapidly worsening progressive myoclonus epilepsy (PME), not treatable with specific therapy. In LBD patients, typical polyglucosan accumulations result from alterations of proteins involved in the regulation of glycogen metabolism. Thus, a ketogenic regimen might reasonably be expected to counteract the disease progression. We set out to assess the feasibility and tolerability of a long-term ketogenic diet (KD) in LBD patients and to make a preliminary evaluation of its effect on the disease course. METHODS: We treated five LBD patients with KD and evaluated the changes in the clinical, neuropsychological and neurophysiological findings over 10-30 months. RESULTS: The KD was well tolerated in all the patients for the first 16 months. Nutritional measures and laboratory findings remained substantially stable. The disease progressed in all the patients, reaching an advanced stage in one. Electrophysiological findings indicated the presence of increased cortical excitability in four patients, paralleling the worsening of the myoclonus. CONCLUSION: KD was unable to stop the disease progression. However, given the considerable heterogeneity of the natural history of LBD, we cannot exclude the possibility that KD has the potential to slow down the disease progression. The application of this nutritional approach should be further evaluated in larger case series.

The ketogenic diet: from molecular mechanisms to clinical effects.

Recent years have witnessed an increased interest from pediatric neurologists, neuropediatricians, epileptologists and general neurologists in the use of the ketogenic diet (KD) for the management of refractory epilepsies, particularly in children and adolescents. This article summarizes current knowledge on various issues related to its use, as discussed at a recent international workshop. Aspects discussed in some detail include (i) the putative mechanisms responsible for the diet's anticonvulsant effects,based on results of biochemical and neurophysiological studies in experimental models; (ii) consensus and controversies on the modalities of initiation of the diet, and different protocols of implementation; (iii) indications and contraindications; (iv) efficacy data, also in relation to seizure type, syndromic form and patients age; (v) adverse effects; (vi) methodological aspects related to assessment of the diet's clinical effects, and perspectives for future research. Overall, the data reviewed indicate that considerable advances have been made in understanding the modes of action of the diet, its efficacy and tolerability profiles and its potential role in different types of epilepsy. Although clinical studies performed to date have important methodological limitations, including suboptimally characterized patients' populations and an uncontrolled design, a number of innovative, prospective randomized study protocols have been recently proposed and are being implemented. The results of these will hopefully provide much needed high-quality information to better define the role of the diet in the treatment algorithms in different epilepsy syndromes.

Psychopathology in children and adolescents with Tourette's syndrome: a controlled study.

OBJECTIVE: Few controlled studies have considered, in paediatric subjects, associations between Tourette's syndrome (TS) and psychiatric/behavioural disorders. We conducted a case-control study to verify the reproducibility of the few data published on this topic to date. METHOD: Clinicians' impression (i.e. structured interviews and usual history taking), standardized psychiatric and behavioural rating scales (CBCL, TAI, CDI, CRS-R:L, Y-GTSS, CY-BOCS) were used to investigate these associations in 17 youngsters with TS and in 17 age- and sex-matched controls. RESULTS: The clinician's diagnoses revealed TS alone in 23.5% of the patients (4/17), TS plus attention-deficit/hyperactivity disorder (ADHD) in 11.8% (2/17), TS plus obsessive-compulsive disorder (OCD) in 41.2% (7/17), and TS plus ADHD and OCD in 23.5% (4/17). No control subject presented ADHD and/or OCD. These clinical findings were confirmed by Child Behaviour Check List (CBCL) results. The TS group, compared with the controls, recorded significantly higher CBCL scores in scales relating to the main comorbid conditions. No significant differences emerged on the Children's Depression Inventory or Test Anxiety Inventory. CONCLUSION: TS patients have a high prevalence of psychiatric and behavioural problems compared with controls. The CBCL is a rapid and useful screening-diagnostic instrument for highlighting the main psychiatric and behavioural problems in TS.

Factors predicting the efficacy of botulinum toxin-A treatment of the lower limb in children with cerebral palsy.

Botulinum toxin A is widely used for spasticity management in children with cerebral palsy, although outcomes are unpredictable. The aim of this study was to identify criteria for selecting patients most likely to benefit from botulinum toxin A treatment. Fifty-five subjects, aged 2.5 to 18 years, were recruited. The assessment covered measures of spasticity (Modified Ashworth Scale), function (using the Gross Motor Function Measure and the Physician's Rating Scale), selective motor control, static range of motion at the ankle with knee extended and flexed, range of motion of the knee flexors, central and peripheral vision, and cognitive ability. Outcomes at 3 months were compared with baseline values. All of the scales showed significant differences between pre- and postinjection values. Significantly increased Gross Motor Function Measure scores were found in children aged 48 months or under and in those able to walk with support. Greater improvements in selective motor control and Physician's Rating Scale were seen in those with a less severe pattern of paralysis, lower levels of impairment, the ability to walk (with or without support), normal visual acuity, and normal or borderline cognition. We identified factors that mark out patients as most likely to achieve functional gains: young age, hemiplegia or diplegia, slight to moderate disability, walking with support, normal or borderline cognition, and normal or borderline visual acuity.

Motor impairment on awakening in a patient with an EEG pattern of "unilateral, continuous spikes and waves during slow sleep".

Movement disorders are rarely described in association with the "continuous spikes and waves during slow sleep (CSWS)" EEG pattern. We report the case of a young girl affected by an epileptic encephalopathy who, from the age of seven years and four months, has twice presented a movement disorder affecting the right arm, manifesting on awakening and disappearing by early afternoon. Sleep EEG during these periods showed continuous, high-amplitude, diphasic spikes and slow waves over the left hemisphere. Association of clobazam, valproic acid and, on the second occasion, ethosuccimide led to disappearance of the above-described EEG picture and associated motor symptoms. Neurophysiological investigations excluded other possible aetiologies. In view of this, and of the close relationship between the EEG picture and clinical course, we interpret the patient's impairment as "motor neglect" secondary to the continuous electrical activity recorded during sleep over the left hemisphere and involving the associative areas. This electrical activity in sleep, may be regarded as a "functional lesion" whose clinical consequences can be correlated with the site of the abnormalities.[Published with video sequences].

Idiopathic headache in children under six years of age: a follow-up study.

OBJECTIVE: To assess the evolution of idiopathic headache with early onset and to investigate the influence of early somatic disorders, "life events," and psychiatric disorders on the onset and the course of headache. We also studied the possible prognostic role of gender, headache diagnosis at onset, and history of headache in family members on the course of headache. We also investigated the applicability of the ICHD-II criteria to idiopathic headache in preschool children. METHODS: Prospective evaluation of 25 consecutive headache patients referred before the age of 6 years. Headache diagnosis was based on the IHS 1988 criteria, on the ICHD-II criteria, and on "alternative" clinical criteria [eg, duration less than 1 hour in migraine without aura (MWA), less than 30 minutes in tension-type headache (TTH)]. All patients were assessed by a structured interview to detect early developmental disorders (eg, feeding difficulties or sleep disorders) and "life events" (eg, parents' separation and others according to DSM-IV). All patients underwent clinical observations and assessment of psychiatric comorbidity (ICD-10) by means of interviews and the Child Behaviour Check List. All the above-mentioned evaluations were performed at recruitment (T0) and at the end of the follow-up period (T1). RESULTS: A total of 25 children with headache (12 males; 13 females) were monitored through long-term clinical follow-up (mean duration: 4.2 years; range: 2.8 to 6.6 years). The "definite" diagnosis of migraine without aura or TTH at T0 was possible in only 6/25 (24%) and 9/25 patients (36%) applying the IHS 1988 criteria and the ICHD-II criteria respectively, but in 20/25 (80%) applying the "alternative" clinical criteria. Evaluation of headache at T1 revealed: remission in 16/25 (64%) patients and persistence in 9/25 (36%). At T1, the ICHD-II diagnosis was possible in 100% of the children with headache persistence (1/9 migraine without aura and 8/9 TTH) and these diagnoses were found to be perfectly concordant with those obtained applying the "alternative criteria." Early developmental disorders were present in 11/25 children, with a significantly higher prevalence in children with headache persistence compared to children showing headache remission (78% vs. 25%; P < .05). No significant differences were found between patients with headache persistence and patients with headache remission with regard to gender, history of headache in family members, headache diagnosis at onset, psychiatric comorbidity at T0, and with regard to "life events" at both T0 and T1. Conversely, the children presenting psychiatric disorders at T1 were more frequently affected by headache than those without psychiatric disorders (59% vs. 15%; P < .05). CONCLUSIONS: Our results suggest that the ICHD-II criteria are too restrictive to allow the classification of migraine without aura and TTH in preschool children. Nevertheless, a diagnosis based on these criteria was possible in all the patients with headache persistence at the end of several years' follow-up. We found a significant association between early somatic disorders and persistence of headache and also between the presence of psychiatric disorders at the end of follow-up and the persistence of headache. "Life events," on the other hand, while not showing a statistically significant association with the evolution of the headache, may nevertheless influence the course of the headache in some patients. Our results suggest that environmental and psychological factors play an important role in idiopathic headache with onset in preschool age, and thus that the diagnostic-therapeutic approach must take these factors into account.

Alternative therapies in the treatment of headache in childhood, adolescence and adulthood.

In recent years, it has become common practice, in the treatment of headache, to use alternative methods, both alone and in association with drug therapies. Alternative therapies would appear to be more indicated in certain subjects: patients opposed to pharmacological treatments, those showing low tolerance of drugs or with a history of drug abuse, and those presenting medical contraindications or poor response to certain drugs. Numerous studies of alternative therapies have been published and here we review the literature data on the topic, considering in particular the accuracy of the various study methods in evaluating the effectiveness of the different therapies and their specificity for the different forms of headache. Specialists involved in the assessment and care of headache patients should strive to increase their knowledge of alternative therapies, so as to be better equipped to guide patients towards safe, economical and potentially effective treatments, rather than useless, costly or dangerous ones.

Facial erythema associated with short stature, absent distal phalanx, dental and nail anomalies: case report and neuropsychological profile.

We describe a patient presenting with a combination of dental agenesis, agenesis of the distal phalanx of the fifth finger, small, dystrophic toenails, permanent facial erythema, short stature and mild mental retardation. The neuropsychological profile of this patient which was studied in detail and showed a significant discrepancy between verbal and performance IQ. We discuss the differential diagnosis of this case and conclude that our patient's condition could represent a new syndrome.

Long-term follow-up of an adolescent who had bilateral striatal necrosis secondary to Mycoplasma pneumoniae infection.

We describe the long-term follow-up of a patient with bilateral striatal necrosis associated with Mycoplasma pneumoniae infection occurred in adolescence. In the literature there are no longitudinal studies of such a rare condition. Our patient, 4 years after the onset of an acute and reversible akinetic-rigid syndrome, showed a severe obsessive-compulsive disorder, cognitive decline, and a neuropsychological profile characterized by signs of deficient executive functioning. The clinical picture that emerged in our patient is suggestive of a frontosubcortical dementia which might be considered a major long-term sequela of the bilateral selective striatal necrosis and consequent dysfunction of frontostriatal connections.

Towards improved clinical characterization of Leber congenital amaurosis: neurological and systemic findings.

Leber congenital amaurosis (LCA) is the most severe form of inherited retinal dystrophy that presents in infancy. LCA is both clinically and genetically heterogeneous. The aim of our study was to clarify the clinical aspects of LCA and to contribute to improved characterization of the disorder. We studied 40 children affected by LCA (mean age at first observation: 19 months, range: 8-50 months), who underwent a comprehensive evaluation that included: neurophthalmological evaluation, electroretinogram (ERG), and visual evoked potentials (VEPs), general and neurological examinations, developmental assessment using scales for visually impaired children, neuroradiological examinations, hepatic and renal function and metabolic investigations, brainstem auditory evoked potentials (BAEPs), EEG, and hand radiographs. Analyses of known LCA genes are ongoing. The subjects are still being followed up at 6-/12-month intervals. All the subjects fulfilled De Laey's criteria for LCA. The neurological examination was abnormal in 31 cases (hypotonia, ataxia with/without associated cerebellar signs). Cognitive development was normal in 24 cases, borderline in five, and subnormal in 11. Mild and nonspecific alterations on MRI were present in seven cases, and "molar tooth" sign in four; all the others had a normal neuroradiological picture. Among the subjects presenting with neurological signs, a subgroup (13 patients) emerged that was characterized by systemic (skin, kidney, liver) involvement. Our data confirm that LCA is a heterogeneous entity that can present as an isolated ocular manifestation, or in association with neurological and systemic abnormalities and support the need for a multidisciplinary approach to this entity and for genotype-phenotype studies.

Visual-perceptual impairment in children with periventricular leukomalacia.

We set out to define visuo-perceptual impairment related to periventricular leukomalacia (PVL) using the Developmental Test of Visual Perception (DTVP). Correlations were sought between visual-perceptual deficits and DTVP profile and neuroradiological and neurophthalmological findings. The DTVP was administered to 20 children (m/f: 10/10), aged between 5 and 8 years (mean: 6.95 years), presenting with: spastic diplegia; PVL documented by brain MRI; normal or mildly impaired visual acuity; mild-moderate upper limb functional impairment. The mean General Visual-Perceptual Quotient was impaired, showing a great variability among the patients. Despite this, an uneven DTPV profile, characterised by a significant difference between the VMIQ and the Non-Motor Visual-Perceptual Quotient (P < 0.001) and a poor result on the Closure subtest (identification of whole figures from incomplete visual information) was observed in all the subjects. This profile reflects a deficit in eye-hand coordination and in praxic-constructional abilities and could be the expression of malfunctioning of the occipital-parietal pathway of visual integration, the so-called 'dorsal stream,' a hypothesis reinforced by the emergence of a statistically significant correlation between the neuroradiological data and the presence of visual-perceptual impairment.