Kentaroh Kuwabara

Identification of a novel biomarker candidate, a 4.8-kDa peptide fragment from a neurosecretory protein VGF precursor, by proteomic analysis of cerebrospinal fluid from children with acute encephalopathy using SELDI-TOF-MS

BackgroundAcute encephalopathy includes rapid deterioration and has a poor prognosis. Early intervention is essential to prevent progression of the disease and subsequent neurologic complications. However, in the acute period, true encephalopathy cannot easily be differentiated from febrile seizures, especially febrile seizures of the complex type. Thus, an early diagnostic marker has been sought in order to enable early intervention. The purpose of this study was to identify a novel marker candidate protein differentially expressed in the cerebrospinal fluid (CSF) of children with encephalopathy using proteomic analysis.MethodsFor detection of biomarkers, CSF samples were obtained from 13 children with acute encephalopathy and 42 children with febrile seizure. Mass spectral data were generated by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) technology, which is currently applied in many fields of biological and medical sciences. Diagnosis was made by at least two pediatric neurologists based on the clinical findings and routine examinations. All specimens were collected for diagnostic tests and the remaining portion of the specimens were used for the SELDI-TOF MS investigations.ResultsIn experiment 1, CSF from patients with febrile seizures (n = 28), patients with encephalopathy (n = 8) (including influenza encephalopathy (n = 3), encephalopathy due to rotavirus (n = 1), human herpes virus 6 (n = 1)) were used for the SELDI analysis. In experiment 2, SELDI analysis was performed on CSF from a second set of febrile seizure patients (n = 14) and encephalopathy patients (n = 5). We found that the peak with an m/z of 4810 contributed the most to the separation of the two groups. After purification and identification of the 4.8-kDa protein, a 4.8-kDa proteolytic peptide fragment from the neurosecretory protein VGF precursor (VGF4.8) was identified as a novel biomarker for encephalopathy.ConclusionsExpression of VGF4.8 has been reported to be decreased in pathologically degenerative changes such as Alzheimers disease, amyotrophic lateral sclerosis (ALS), frontotemporal dementia, and encephalopathy. Thus, the VGF4.8 peptide might be a novel marker for degenerative brain conditions.

Acute pancreatitis with Kawasaki disease: analysis of cases with elevated serum amylase levels

Kawasaki disease (KD) is a multi-system disorder of unknown origin [4]. Although inflammatory changes in the pancreas have been reported in KD [1, 6], there have been few reports of clinical pancreatitis [2, 3, 5]. A boy (aged 1 year and 8 months) was admitted to our hospital as having KD. Five days before admission, he was diagnosed with an influenza A infection and was prescribed oseltamivir for 5 days. Slightly increased levels of serum amylase (193 IU/; reference range 80– 170 IU/l) with predominant pancreatic amylase isozyme (pancreas 90% versus salivary gland 10%; reference range 21%–65% versus 35%–79%), pancreas secretory trypsin inhibitor (138.6 ng/ml; reference range 4.2– 12.2 ng/ml), elastase 1 (525 ng/dl; reference range 100– 400 ng/dl), lipase (89 IU/l; reference range 11–53 IU/l), and pancreatic phospholipase A2 (861 ng/dl; reference range 130–400 ng/dl) were noted with swelling of the pancreas. As he exhibited no symptoms suggestive of pancreatitis, he was not treated for severe pancreatitis and he received intravenous gamma globulin therapy

Seronegative Antiphospholipid Syndrome with Anti-phosphatidylethanolamine Antibody in a Boy.

Antiphospholipid syndrome (APS) is an autoimmune disease caused by antiphospholipid antibodies. At our institution, APS is diagnosed on the basis of the Sapporo criteria, which consist of thrombosis and recurrent pregnancy-related complications and the following laboratory findings: the presence of lupus anticoagulant, anticardiolipin antibody, or anti-β2 glycoprotein 1 antibody. However, we sometimes treat patients we strongly suspect of having APS but who do not satisfy the laboratory criteria. To accommodate such suspected cases, a subtype of APS termed seronegative APS has been proposed. Here, we report on a man with chronic thromobocytopenic purpura since the age of 3 years and multiple cerebral infarctions since the age of 14 years who finally received a diagnosis of seronegative APS with positive antiphosphatidylethanolamine antibodies.

Neurocutaneous melanosis with acute disseminated encephalomyelitis

Neurocutaneous melanosis (NCM) is a rare congenital disease characterized by the presence of large or multiple congenital melanocytic nevi and benign or malignant melanocytic tumors of the leptomeninges [2] with high risk of melanoma development [1]. Acute disseminated encephalomyelitis (ADEM) is an uncommon monophasic inflammatory demyelinating disease that usually presents in children and young adults [3]. Here, we report a boy aged 10 years and 7 months with NCM complicated by ADEM.

Usefulness of DNA analysis of tuberculosis with ruptured lymphadenitis in a bacille Calmette–Guérin‐vaccinated infant

However, in 0.02% of vaccinated children, the disease progresses to suppurative adenitis with rupture, after which a decision must be made regarding whether anti-tuberculosis medication should be started to minimize bacterial dissemination. 1 Mycobacterium bovis BCG is genetically and phenotypically similar to other strains of M. bovis and other species of the M. tuberculosis complex ( M. tuberculosis , M. africanum , and M. microti ). 3 Because there is no defi nitive test to differentiate between these strains, even using a combination of biochemical and growth features, the best course of treatment is not established. Polymerase chain reaction (PCR) methods have recently been utilized in differential diagnoses for several microorganisms, including M. tuberculosis . 3 – 5 Here, we report axillary suppurative lymphadenitis with rupture in a BCG-vaccinated infant. The isolate was confi rmed to be of BCG origin on PCR and restriction fragment length polymorphism (RFLP), in addition to standard culture and biochemical methods, and these fi ndings greatly assisted in deciding the treatment schedule.

Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome in a 14-year-old boy: an immunohistochemical study of infiltrating lymphocytes in acneous skin regions.

Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome was proposed as a clinico-radiological entity combining skin, bone and joint manifestations [1]. Here, we report a 14-year-old boy with SAPHO syndrome, in whom we investigated the pathology of acneous skin regions with aberrant T-cell lymphocyte infiltration in the dermal area. A 14-year-old boy was admitted to our hospital exhibiting persistent high-grade fever with acne conglobata, in addition to chest, left knee joint and lumbar pain. Physical examination revealed swelling of the left knee joint and severe facial acne (Fig. 1). Haematological and biochemical investigations were all normal; however, C-reactive protein was 8.58 mg/dl and the ESR was 54 mm/h. Urinalysis showed normal findings. The purified protein derivative of tuberculosis reaction was unchanged from the previous year. Blood, urine, stool and pharyngeal cultures were all negative. Compliment, ferritin, and serum immunoglobulin level were normal. LE test, antibodies related to collagen diseases and tumour markers were all negative. Gallium scintigraphy showed abnormal accumulation in the left iliosacral joint, bilateral parotid glands, and acne lesions of the forehead (Fig. 2). CT scans of the pelvis showed increased space in the left iliosacral joint, thus suggesting swelling of the joint cartilage, but no solid-type periosteal reaction [6] were observed (Fig. 3). After admission, we treated the patient with intravenous antibiotics, but the clinical symptoms remained unchanged. For differential diagnosis, we performed a skin biopsy and found pyoderma with focal foreign body reaction and aberrant lymphoid cell infiltration (Fig. 4). We then started to administer oral ibuprofen. Clinical symptoms and laboratory findings improved rapidly after ibuprofen administration. Two years after discharge, his course has been uneventful with continued oral administration of ibuprofen. Because our case has not been refractory, we did not use pamidronate [4].