CHIPS for genetic testing to improve a regional clinical genetic service.

In current practice of clinical genetics, molecular diagnosis has become more widely used than ever before. DNA diagnosis is important for appropriate medical care of the patient, and proper genetic counseling to the family. However, genetic testing of orphan disease cannot always be performed easily. In multiple congenital anomalies (MCA) syndromes by monogenic cause, the broad mutational spectrum and large size of responsible genes often make molecular diagnosis expensive and cumbersome. We solve this problem with on-demand genetic testing by CHIPS (CEL nuclease mediated heteroduplex incision with polyacrylamide gel electrophoresis and silver staining) technology, which is the ultimately conventional and economical mutation screening system. In this article, we show eight patients with MCA syndromes who were recently treated at our hospital, and demonstrate that CHIPS successfully offers efficient and inexpensive genetic testing and facilitates clinical genetic service in our local region.

SANASA Capivari II - the first full-scale municipal membrane bioreactor in Latin America.

The macro region of Campinas (Brazil) is rapidly evolving with new housing developments and industries, creating the challenge of finding new ways to treat wastewater to a quality that can be reused in order to overcome water scarcity problems. To address this challenge, SANASA (a publicly owned water and wastewater concessionaire from Campinas) has recently constructed the 'EPAR (Water Reuse Production Plant) Capivari II' using the GE ZeeWeed 500D(®) ultrafiltration membrane system. This is the first large-scale membrane bioreactor (MBR) system in Latin America with biological tertiary treatment capability (nitrogen and phosphorus removal), being able to treat an average flow of 182 L/s in its first phase of construction. The filtration system is composed of three membrane trains with more than 36,000 m(2) of total membrane filtration area. The membrane bioreactor (MBR) plant was commissioned in April 2012 and the permeate quality has exceeded expectations. Chemical oxygen demand (COD) removal rates are around and above 97% on a consistent basis, with biochemical oxygen demand (BOD5) and NH3 (ammonia) concentrations at very low levels, and turbidity lower than 0.3 nephelometric turbidity unit (NTU). Treated effluent is sent to a water reuse accumulation tank (from where will be distributed as reuse water), and the excess is discharged into the Capivari River.

SCN1B is not related to benign partial epilepsy in infancy or convulsions with gastroenteritis.

We hypothesized that benign partial epilepsy in infancy (BPEI) and convulsions with gastroenteritis (CwG) may have a similar genetic background, because previous studies indicate that clinical features overlap between BPEI and CwG. As carbamazepine is effective for cessation of clustering seizures in children with BPEI and CwG, some genetic mutations regarding sodium channels may be related to the development of BPEI and/or CwG. We focused on SCN1B encoding the voltage-dependent sodium channel ß subunit. We explored SCN1B mutation in 6 children with BPEI and 6 children with CwG. Genomic DNAs were extracted from peripheral blood samples accumulated from the patients and all 5 exons of SCN1B were amplified by standard PCR amplification. There were no SCN1B mutations or pathological single nucleotide polymorphisms in any of the patients, although the phenotypes of our patients were typical for BPEI or CwG. Our study demonstrated that SCN1B may not be related to the occurrence of BPEI or CwG.

Transient and mild reduction of consciousness during febrile illness in children.

We have studied the clinical and neuroimaging characteristics of transient and mild reduction of consciousness during febrile illness in children. We retrospectively evaluated 58 children admitted with mild reduction of consciousness within 12 h during febrile illness. 53 patients (91%) had delirious behavior, and 5 (9%) had no delirious behavior. We also compared the clinical characteristics, brain magnetic resonance imaging (MRI) findings, and electroencephalography (EEG) findings between patients with and without delirious behavior, and no statistically significant differences were observed in any of them between the 2 patient groups (P≥0.05). MRI was performed 0-4 days after onset in 23 patients. Reversible splenial or callosal and white matter lesions were observed in 2 of 3 patients without delirious behavior vs. 4 of 20 patients with delirious behavior on diffusion-weighted images. EEG was performed 0-3 days after onset in 29 patients. Transient abnormal findings were observed in 3 of 4 patients without delirious behavior vs. 11 of 25 patients with delirious behavior. In conclusion, we consider that transient and mild reduction of consciousness during febrile illness is a unique clinical group that is constituted by children both with and without delirious behavior.

Acute encephalopathy in a patient with Dravet syndrome.

Dravet syndrome (severe myoclonic epilepsy in infancy) is an epileptic syndrome with various types of seizures that begin in the first year of life and may result in intellectual impairment. Mutations of the SCN1A gene are the most prevalent genetic cause of Dravet syndrome. In this study, we report a 12-year-old girl with Dravet syndrome carrying an SCN1A mutation, c.2785Cdel (L929del fsX934). She had an episode of status epilepticus and persistent lethargy after 48 h of acute febrile illness that was preceded by an annual flu vaccination. Low voltage activities detected by electroencephalogram and elevated neuron-specific enolase/interleukin-6 concentrations in the cerebrospinal fluid suggested acute encephalopathy. MRI showed abnormalities in the bilateral thalami, cerebellum and brainstem. These abnormalities were protracted over a month. The biochemical and MRI characteristics of this case are different from any known type of encephalopathy, and may suggest a vulnerability of neurons expressing mutant SCN1A in the brain.

UK B.1.1.7 variant exhibits increased respiratory replication and shedding in nonhuman primates.

The continuing emergence of SARS-CoV-2 variants calls for regular assessment to identify differences in viral replication, shedding and associated disease. In this study, African green monkeys were infected intranasally with either a contemporary D614G or the UK B.1.1.7 variant. Both variants caused mild respiratory disease with no significant differences in clinical presentation. Significantly higher levels of viral RNA and infectious virus were found in upper and lower respiratory tract samples and tissues from B.1.1.7 infected animals. Interestingly, D614G infected animals showed significantly higher levels of viral RNA and infectious virus in rectal swabs and gastrointestinal tract tissues. Our results indicate that B.1.1.7 infection in African green monkeys is associated with increased respiratory replication and shedding but no disease enhancement similar to human B.1.1.7 cases. ONE-SENTENCE SUMMARY: UK B.1.1.7 infection of African green monkeys exhibits increased respiratory replication and shedding but no disease enhancement.

Does the pediatricians' work setting or years of experience influence febrile seizure education?

INTRODUCTION: A febrile seizure is a benign condition. However, for parents, witnessing their child's FS can cause excessive anxiety. It is therefore important for pediatricians to provide appropriate information in order to reduce anxiety. In this study, we analyze whether work setting and years of experience influence the explanations given to caregivers. METHODS: Questionnaires were mailed to members of the Japan Pediatric Society, Tokyo Chapter (n=1 870). The Mantel-Haenszel test was used for dichotomous variables. Differences for continuous variables were evaluated at 95% confidence intervals. RESULTS: A total of 482 pediatricians participated. There were no significant differences in responses to any questions according to work setting. Responders with less than 20 years of experience reported a higher prevalence of febrile seizures than those in the more experienced group. Compared to the experienced group, more responders with less than 20 years of experience stated that they would administer antiepileptic prophylaxis and advise parents not to use antipyretics, and indicated that they did not know the FS treatment guidelines. CONCLUSION: The findings suggest the importance of promoting a better understanding of FS among less-experienced pediatricians and encouraging adherence to the guidelines to maintain a consistent level of support for parents and caregivers.

Congenital infiltrating lipomatosis of the face with ipsilateral hemimegalencephaly, band heterotopia, and hypertrophy of brainstem and cerebellum.

The simultaneous appearance of congenital infiltrating lipomatosis of the face that causes facial hemihypertrophy and ipsilateral hemimegalencephaly is extremely rare. We report a 4-year-old boy with congenital facial asymmetry and infantile-onset epilepsy. Magnetic resonance imaging (MRI) results led to the diagnosis of infiltrating lipomatosis of the face; the diagnosis was confirmed on the basis of the results of pathological examinations. Additionally, brain MRI revealed ipsilateral hemimegalencephaly, associated with band heterotopia and the hemihypertrophy of the ipsilateral brainstem and cerebellum. He had no nevi or other skin abnormalities suggesting neurocutaneous syndrome. His seizures were so intractable that they necessitated functional hemispherectomy. The lipomatous lesion was successfully resected without relapse. Psychomotor delay and left hemiplegia were observed at the last follow-up.

Absent cyclicity on aEEG within the first 24 h is associated with brain damage in preterm infants.

OBJECTIVE: Our aim was to clarify the relationship between amplitude-integrated electroencephalographic (aEEG) findings before 24 h of age in preterm infants and neurodevelopmental outcome. DESIGN: 12 infants born between 27 and 32 weeks of gestation were eligible. The recordings of aEEG and conventional EEG were started within 12 h after birth. The background aEEG findings were evaluated and classified. Additionally, we evaluated the absence or presence of changes on the lower border of the aEEG. RESULTS: All infants had discontinuous normal voltage background on aEEG, corresponding to decreased or normal continuity on conventional EEG. Cyclicity on aEEG was seen in 8 of 12 infants within 24 h of age, and all of these infants had favourable outcomes. Cyclicity on aEEG was not recognized in 4 infants. 3 of the 4 infants with absent cyclicity had abnormal neurodevelopmental outcomes at 12 months. One of these infants had intraventricular haemorrhage (grade 2) with delayed development, and 2 had cystic periventricular leukomalacia followed by spastic diplegia. CONCLUSION: Absent cyclicity on aEEG within 24 h of age was associated with poor outcome in preterm infants.

An African pygmy hedgehog adenovirus 1 (AhAdV-1) outbreak in an African pygmy hedgehog (Atelerix albiventris) colony in Japan.

An African pygmy hedgehog adenovirus 1 (AhAdV-1) outbreak in a colony of 24 African pygmy hedgehogs (APHs) with a case of fatal pneumonia occurred in Japan. Thirteen out of a colony of 15 APHs with respiratory symptoms were diagnosed with AhAdV-1 infection based on the detection of AhAdV-1 genome in throat/nasal swabs and further one APH was diagnosed on isolation of the virus. Five infected APHs died during the outbreak and AhAdV-1 caused severe pneumonia and death in one case. After the outbreak, persistent AhAdV-1 infection was suggested in one surviving APH. AhAdV-1 is a novel adenovirus and is suspected to be an emerging pathogen.

A rare case of fetal extensive intracranial hemorrhage and whole-cerebral hypoplasia due to latent maternal vitamin K deficiency.

We present here a late preterm infant with extensive brain lesions resulting from vitamin K deficiency. A female infant was born after 35 weeks of gestation by emergent cesarean section because of non-reassuring fetal status. Her mother had severe eating disorder and recurrent vomiting since early pregnancy. She was immediately intubated and ventilated because she was extremely pale, hypotonic, and non-reactive. Cerebral magnetic resonance imaging immediately after birth showed intraparenchymal hemorrhage in the left frontal lobe and cerebellum, marked cerebral edema, and cerebellar hypoplasia. Coagulation studies of the infant showed hepaplastin test <5%, prolonged PT and APTT, and a marked elevation of protein induced by vitamin K absence or antagonist-II. This case highlighted a potential risk of intracranial bleeding due to maternal vitamin K deficiency and difficulty in its prediction before delivery. Vitamin K supplementation to high risk mothers might be indispensable for preventing severe fetal vitamin K deficiency. Even when coagulation studies in mothers is normal, it is imperative to provide vitamin K supplementation for total protection.

Focal brain glucose hypometabolism in patients with neuropsychologic deficits after diffuse axonal injury.

BACKGROUND AND PURPOSE: Neuropsychologic deficits are well-known sequelae of traumatic brain injury. However, the cerebral correlates of these deficits are still unclear. The aim of the present study was to elucidate the regions of cerebral dysfunction correlated with such neuropsychologic deficits after traumatic brain injury. METHODS: Sets of fluorine-18 fluorodeoxyglucose-positron-emission tomography (FDG-PET) images in the resting state were obtained from 12 patients with neuropsychologic deficits after diffuse axonal injury and from 32 healthy volunteers. The cortical metabolic activity of each subject's PET image sets was extracted using 3D stereotactic surface projection (3D-SSP). A "normal" data base was created using the extracted datasets of the healthy subjects. The patients' datasets were compared with the normal data base by calculating a statistical Z-score on a pixel-by-pixel basis in searches for focal metabolic abnormalities. RESULTS: Group comparisons revealed hypometabolism in the cingulate gyrus with additional involvement of the lingual gyrus and cuneus. Individual case-by-case analyses disclosed differences in the site and extent of the hypometabolism in the cingulate gyrus of each case. Predominant hypometabolism was found in the anterior cingulate gyrus of 6 patients, the middle cingulate gyrus of 2 patients, and the posterior cingulate gyrus of 4 patients. CONCLUSION: Interpretation of FDG-PET using 3D-SSP facilitates the identification of regional hypometabolism in the cerebral cortex of patients after diffuse axonal injury. Dysfunction of the cingulate gyrus, lingual gyrus, and cuneus may play a crucial role in neuropsychologic deficits after traumatic brain injury.

Predominant area of brain lesions in neonates with herpes simplex encephalitis.

OBJECTIVE: Nonspecific manifestations and a varied distribution of brain lesions can delay the diagnosis of herpes simplex encephalitis (HSE) in neonates. The aim of this study was to report predominant brain lesions in neonatal HSE, and then to investigate the association between pattern of predominant brain lesions, clinical variables and neurodevelopmental outcome. STUDY DESIGN: A multicenter retrospective study was performed in neonates diagnosed with HSE between 2009 and 2014. Magnetic resonance (MR) images, including diffusion-weighted images, were obtained in the acute and chronic phase. RESULTS: Three predominant areas of brain injury could be defined based on characteristic MRI findings in 10 of the 13 infants (77%). The inferior frontal/temporal pole area was involved in five (38%) patients. The watershed distribution was present in six (46%) patients. Four (31%) infants involved the corticospinal tract area. No significant association was found between any predominant distribution of brain lesion pattern and sex, country, viral type or viral load. However, the corticospinal tract involvement was significantly associated with motor impairment (P=0.045). CONCLUSION: Three predominant areas of brain lesion could be recognized in neonatal HSE. Recognition of those areas can improve prediction of neurodevelopmental outcome.

Relationship between regional cerebral metabolism and consciousness disturbance in traumatic diffuse brain injury without large focal lesions: an FDG-PET study with statistical parametric mapping analysis.

BACKGROUND: The cerebral metabolism of patients in the chronic stage of traumatic diffuse brain injury (TDBI) has not been fully investigated. AIM: To study the relationship between regional cerebral metabolism (rCM) and consciousness disturbance in patients with TDBI. METHODS: 52 patients with TDBI in the chronic stage without large focal lesions were enrolled, and rCM was evaluated by fluorine-18-fluorodeoxyglucose positron emission tomography (FDG-PET) with statistical parametric mapping (SPM). All the patients were found to have disturbed consciousness or cognitive function and were divided into the following three groups: group A (n = 22), patients in a state with higher brain dysfunction; group B (n = 13), patients in a minimally conscious state; and group C (n = 17), patients in a vegetative state. rCM patterns on FDG-PET among these groups were evaluated and compared with those of normal control subjects on statistical parametric maps. RESULTS: Hypometabolism was consistently indicated bilaterally in the medial prefrontal regions, the medial frontobasal regions, the cingulate gyrus and the thalamus. Hypometabolism in these regions was the most widespread and prominent in group C, and that in group B was more widespread and prominent than that in group A. CONCLUSIONS: Bilateral hypometabolism in the medial prefrontal regions, the medial frontobasal regions, the cingulate gyrus and the thalamus may reflect the clinical deterioration of TDBI, which is due to functional and structural disconnections of neural networks rather than due to direct cerebral focal contusion.

Evidence for white matter disruption in traumatic brain injury without macroscopic lesions.

BACKGROUND: Non-missile traumatic brain injury (nmTBI) without macroscopically detectable lesions often results in cognitive impairments that negatively affect daily life. AIM: To identify abnormal white matter projections in patients with nmTBI with cognitive impairments using diffusion tensor magnetic resonance imaging (DTI). METHODS: DTI scans of healthy controls were compared with those of 23 patients with nmTBI who manifested cognitive impairments but no obvious neuroradiological lesions. DTI was comprised of fractional anisotropy analysis, which included voxel-based analysis and confirmatory study using regions of interest (ROI) techniques, and magnetic resonance tractography of the corpus callosum and fornix. RESULTS: A decline in fractional anisotropy around the genu, stem and splenium of the corpus callosum was shown by voxel-based analysis. Fractional anisotropy values of the genu (0.47), stem (0.48), and splenium of the corpus callosum (0.52), and the column of the fornix (0.51) were lower in patients with nmTBI than in healthy controls (0.58, 0.61, 0.62 and 0.61, respectively) according to the confirmatory study of ROIs. The white matter architecture in the corpus callosum and fornix of patients with nmTBI were seen to be coarser than in the controls in the individual magnetic resonance tractography. CONCLUSIONS: Disruption of the corpus callosum and fornix in patients with nmTBI without macroscopically detectable lesions is shown. DTI is sensitive enough to detect abnormal neural fibres related to cognitive dysfunction after nmTBI.

Parry-Romberg syndrome with a clinically silent white matter lesion.

We performed a detailed neuroimaging study in a patient with Parry-Romberg syndrome. Proton MR spectroscopy demonstrated normal spectral patterns, though conventional MR imaging revealed high-intensity areas in the entire white matter in the left hemisphere. Single-photon emission tomography showed increased perfusion in the cortex of the affected hemisphere. Pyramidal tracts and optic radiations were preserved on diffusion tensor tractography. We will correlate these neuroimaging findings with normal psychomotor development in our patient.

ISG15 enhances the innate antiviral response by inhibition of IRF-3 degradation.

The transcription factor, interferon regulatory Factor 3 (IRF-3) plays a critical role in the activation of an antiviral innate immune response. However the transcriptional activity of IRF-3 is tightly regulated by a proteosome mediated degradation. We describe here a novel mechanism by which the activity of IRF-3 is stabilized in infected cells. We have shown that both interferon treatment and NDV infection profoundly increase conjugation of interferon induced ubiquitin- like protein ISG15 to cellular proteins. ISGylated IRF-3 could be detected both in interferon treated and virus-infected cells. ISG15, subverts the ubiquitin mediated degradation of IRF-3 in NDV infected 2fTGH cells and enhances the NDV mediated transactivation of interferonbeta promoter and the translocation of activated IRF-3 to the nucleus. The relative levels of IRF-3 were significantly lower in NDV infected ISG15 null MEF, than in wt MEF. While ISG15 null MEF were more permissive to VSV replication their sensitivity to the antiviral effect of interferon was not modulated. These results reveal that virus mediated subversion of the antiviral response by proteolysis of IRF-3 is counteracted by induction of ISG15 expression and that ISGylation provides a feedback mechanism, which enhances the host innate antiviral response via IRF-3 stabilization.

Ligands for peroxisome proliferator-activated receptors alpha and gamma inhibit chemically induced colitis and formation of aberrant crypt foci in rats.

The biological role of the peroxisome proliferator-activated receptors (PPARs) in various diseases, including inflammation and cancer, has been highlighted recently. Although PPARgamma ligands have been found to inhibit mammary carcinogenesis in rodents, the effects on colon tumorigenesis are controversial. In the present study, three different experiments were conducted to investigate the modifying effects of PPARs ligands (PPARalpha and PPARgamma) on colitis and an early phase of colitis-related colon carcinogenesis in male F344 rats. In the first experiment, gastric gavage of troglitazone (PPARgamma ligand, 10 or 100 mg/kg body weight) or bezafibrate (PPARalpha ligand, 10 or 100 mg/kg body weight) inhibited colitis induced by dextran sodium sulfate (DSS) and lowered trefoil factor-2 content in colonic mucosa. In the second experiment, dietary administration (0.01 or 0.05% in diet) of troglitazone and bezafibrate for 4 weeks significantly reduced azoxymethane (AOM, two weekly s.c. injections, 20 mg/kg body weight)-induced formation of aberrant crypts foci, which are precursor lesions for colon carcinoma. In the third experiment, dietary administration (0.01% in diet for 6 weeks) of pioglitazone (PPARgamma ligand), troglitazone, and bezafibrate effectively suppressed DSS/AOM-induced ACF. Administration of both ligands significantly reduced cell proliferation activity in colonic mucosa exposed to DSS and AOM. Our results suggest that synthetic PPARs ligands (PPARalpha and PPARgamma) can inhibit the early stages of colon tumorigenesis with or without colitis.