The loss of p53 function is a key event in tumorigenesis. Inactivation of p53 in primary tumors and cell lines is mediated by several molecular mechanisms, including deletions and rearrangements. However, generation of a p53 fusion gene has not yet been reported. Here we report a novel p53/an autosomal homolog of the fragile X mental retardation (FXR2) chimeric gene generated by an interstitial deletion. Western blot analyses have shown that the p53/FXR2 protein is indeed expressed in a Down syndrome-related acute megakaryoblastic leukemia cell line, CMK11-5 cells. To investigate the properties of the p53/FXR2 protein, we observed its subcellular localization. Flag-tagged expression vectors were transfected into COS-7 cells and the proteins were stained with an anti-Flag antibody. The p53/FXR2 protein was expressed at high levels in the cytoplasm, whereas wild-type p53 and FXR2 were localized primarily in the nucleus and in the periphery of the nucleus, respectively. Treatment with a topoisomerase II inhibitor, VP16, failed to induce expression of a p53 target gene, the cyclin-dependent kinase inhibitor p21WAF-1/CIP1, in CMK11-5 cells, and transient transfection analysis showed that the p53/FXR2 protein failed to transactivate the p21WAF-1/CIP1 promoter. These results suggest that the p53/FXR2 fusion protein lacks the ability of wild-type p53 to function as a transcription factor. The p53/FXR2 gene is the first reported p53 fusion gene.
Aspiration of acid to the airway causes airway inflammation, and acid stress to the airway caused by gastroesophageal reflux disease (GERD) has been known as a potential mechanism of deteriorated asthma symptoms. However, the efficacy of the acid suppressive drugs, H2-receptor blockers (H2 blocker) and proton pump inhibitors, on asthma symptoms and pulmonary functions remains controversial. We therefore designed the randomized prospective study to determine the efficacy of an H2 blocker (roxatidine, 150 mg/day) and a proton pump inhibitor (lansoprazole, 30 mg/day) on asthma symptoms of 30 asthmatic patients with GERD. These patients were divided in the two groups (15 patients for each group) and treated with either roxatidine or lansoprazole. The diagnosis of GERD was established by the method of Los Angeles classification including mucosal minimum change of Grade M and questionnaire for the diagnosis of reflux disease (QUEST) score. The efficacy of acid suppressive drugs was evaluated by peak expiratory flow (PEF), asthma control questionnaire (ACQ) that evaluates the improvement of asthma symptoms, and forced expiratory volume in 1 second (FEV1.0). Lansoprazole, but not roxatidine, significantly improved PEF and ACQ scores (p < 0.05) with the improved QUEST scores. However, these acid suppressive drugs did not change the pulmonary function of FEV1.0 in asthmatic patients. In conclusion, treatment with a proton pump inhibitor, lansoprazole, appears to be useful in improvement of asthma symptoms in asthmatic patients with GERD.
Long-term treatment of childhood nephrotic syndrome (NS) and rheumatic diseases with cyclosporine A (CsA) given as a single daily dose may yield better results and allow safer use of the drug than the conventional twice-daily dosing. However, the safety of such long-term treatment from the histological standpoint remains to be established. Posttreatment renal biopsy was conducted in a total of eight children (5 with minimal change NS, 2 with focal segmental glomerulosclerosis and 1 with X-linked immune dysregulation, polyendocrinopathy and enteropathy) receiving CsA as a single daily dose, after a mean treatment duration of 20 months (9-36 months). The initial daily dose of CsA (Neoral) was 2.0 mg/kg, given as a single daily dose before breakfast. The dose was subsequently adjusted to achieve a peak (between 1 and 2 hrs post-dosing, C1-C2) blood level of around 800 ng/ml. The mean daily CsA dose, mean C1-C2 blood level, and mean trough blood level in the subjects were 1.9 ± 0.6 mg/kg, 803.8 ± 117.2 ng/ml and 36.1 ± 12.7 ng/ml, respectively. The result revealed no evidence of CsA-related nephrotoxicity, including arteriopathy, striped interstitial fibrosis or tubular atrophy, in any of the study participants. Also, no significant changes were observed in the mean estimated glomerular filtration rate as compared to the pretreatment values (127.6 ± 14.9 ml/min/1.73 m2 vs 115.6 ± 22.8 ml/min/1.73 m2), and except for mild hypertrichosis, no significant adverse effects of CsA were observed. These findings lend further support to the safety of long-term low-dose CsA treatment (median treatment duration in this study, 20 months), with the drug administered as a single daily dose while maintaining a peak (C1-C2) blood level of around 800 ng/ml.
Prospective studies have demonstrated that markers of inflammation, such as high-sensitivity C-reactive protein (hsCRP) and fibrinogen, predict future cardiovascular disease risk. However, the association between the hsCRP and fibrinogen levels and the extent of coronary stenosis in patients with coronary artery disease (CAD) remains controversial. The aim of our case-control study was to assess the association of inflammatory markers with the occurrence and extent of CAD. Serum hsCRP and plasma fibrinogen levels were measured in 138 patients with angiographically assessed CAD and in 183 healthy subjects matched according to age and gender. According to the number of significantly stenosed (≥ 50%) vessels, the patients were classified in four groups: those without stenosis (0-vessel disease) and those with 1, 2 or 3-vessel disease. The hsCRP and fibrinogen levels were significantly higher in patients than in controls (p < 0.001). Although the hsCRP and fibrinogen levels tended to increase with the number of stenotic vessels, the differences were only significant for hsCRP (p < 0.01). Regression analysis indicated hsCRP as an independent predictor for the presence (OR = 3.573, p < 0.05) and extent of CAD (β = 1.095, p < 0.05). In conclusion, the present study is the first report concerning the frequency distribution of hsCRP in Serbian healthy subjects and CAD patients. We have shown that elevated levels of hsCRP are associated with the presence and extent of CAD.
Invasive fungal infection is a fatal complication in liver transplantation and it is very difficult to diagnose at the early stage. The aim of this study was to review our experience with invasive fungal infections in living donor liver transplantation (LDLT) and to analyze the risk factors and the impact of β-D glucan. From 1991 to 2005, 96 LDLTs were performed in our institution and we measured the serum level of β-D glucan in order to clarify the diagnosis. Invasive fungal infection was diagnosed based on clinical symptoms, culture, radiological evidence and β-D glucan. Active fungal infection was treated with fluconazole, amphotericin B, flucytosine and micafungin. Risk factors both pre- and post- LDLT were analyzed. Candida albicans was the most frequently isolated species (70%). The risk factors identified by univariate analysis include the following four conditions: acute blood purification (plasma exchange with or without continuous hemodiafiltration), hepatic vein complications, renal failure and respiratory failure. By logistic regression analysis, hepatic vein complications and respiratory failure were identified as independent risk factors. The risk factors for invasive fungal infection of LDLT in Japan have not been well analyzed and this report will provide valuable information for the prevention of the fungal infection.
Resistance to cytosine arabinoside (Ara-C) is a major problem in the treatment of patients with acute myeloid leukemia (AML). In order to investigate the mechanisms involved in Ara-C resistance, the gene expression profile of Ara-C-resistant K562 human myeloid leukemia cells (K562/AC cells) was compared to that of Ara-C-sensitive K562 cells (K562 cells) by using a cDNA microarray platform. Correspondence analysis demonstrated that insulin-like growth factor I (IGF-I) gene was upregulated in K562/AC cells. The biological significance of IGF-I overexpression was further examined in vitro. When K562 cells were incubated with IGF-I ligand, they were protected from apoptosis induced by Ara-C. In contrast, a significant inhibition of growth and increase of apoptosis of K562/AC cells were induced by IGF-I receptor neutralizing antibody, or suramin, a nonspecific growth factor antagonist. Moreover, from the analysis of 27 AML patients, we have shown that IGF-I expression levels are higher in patients at refractory stage, after Ara-C combined chemotherapy, than those in patients at diagnosis. These results suggest that the inhibition of IGF-I and its downstream pathway is a valuable therapeutic approach to overcome Ara-C resistance in AML.
Cytotoxin-associated gene A (cagA) of Helicobacter pylori (H. pylori) encodes a highly immunogenic and virulence-associated protein. The presence of cagA+H. pylori strains in tonsil and adenoid tissues may affect clinical outcome. The aim of the present study was to determine the presence of H. pylori cagA gene in tonsil and adenoid tissues and to establish the potential association of cagA+H. pylori in recurrent adenotonsillitis (RAT) and adenotonsillar hypertrophy (ATH). For this aim, a total of 118 tissue samples (71 tonsil and 47 adenoid tissues) were collected from a total of 71 children: 28 cases with RAT and 43 cases with ATH. The samples were analyzed for glmM gene to detect the infection with H. pylori by polymerase chain reaction (PCR). H. pylori-positive samples were further analyzed for the presence of the cagA gene. The PCR analysis showed that 29 samples (24.6%) were positive for H. pylori. Seventeen out of these 29 samples (58.6%) were found positive for cagA; the cagA gene was detected in 12 samples of ATH and 5 samples of RAT. The presence rate of cagA gene was significantly higher (p < 0.05) in ATH patients than that found in RAT patients. These results suggest that presence of cagA+H. pylori may be associated with development of ATH.
Visual evoked potential (VEP) testing is used frequently and is an important ophthalmologic physiological test to examine visual functions objectively. The VEP is a complicated waveform consisting of negative waveform named N75 and N135, and positive waveform named P100. Delayed P100 latency and greatly attenuated amplitude on VEP are known characteristics for diagnosing optic nerve disease. Acupuncture has been used to treat wide clinical symptoms with minimal side effects. The confirmation of the efficacy of acupuncture generally relies on subjective symptoms. There is not much scientific evidence supporting the acupuncture treatments for eye diseases up to today. However, the VEP test can evaluate objectively and numerically the efficacy of the treatment by the acupuncture. We analyzed 19 healthy subjects (38 eyes). The P100 latencies in the group of less than 101.7 msec (total average) before acupuncture stimulations were not different than those after treatment (98.2 ± 3.0 msec, 98.2 ± 4.0 msec, respectively, p = 0.88, n = 17), but the latencies in those subjects with longer or equal to 101.7 msec were statistically different after acupuncture (104.6 ± 2.8 msec, 101.9 ± 3.7 msec, respectively, p = 0.006, n = 21). These results show that the acupuncture stimulation contributes to the P100 latencies of pattern reversal (PR)-VEP to some subjects who have delayed latencies, and this electrophysiological method is a valuable technique in monitoring the effectiveness of acupuncture therapy in the improvements of visual functions. The purpose of this study is to evaluate the physiological effects by acupuncture stimulations using PR-VEP in normal subjects.
Only scarce data are available on chronic copper poisoning in general toxicology literature. This paper reports four patients with chronic copper poisoning and one patient with acute poisoning. The cases with chronic poisoning in our study consisted of four members of a farmer family presenting to the emergency department (ED) with malaise, weakness, abdominal pain, headache, dizziness, tightness in the chest, leg and back pain, accompanied by significant anemia (hemoglobin [Hb]: 8.7 - 9.5 g/dl). They were hospitalized and investigated thoroughly, although there were no other findings or clues enlightening the etiology of anemia. The anemia was attributed to chronic copper exposure acquired from vegetables containing copper. The diagnosis was established by ruling out other possible etiologies and history coupled with laboratory findings. The patients were discharged with the recommendation on diet to avoid consumption of pesticide-treated vegetables. Their Hb values were between 10 and 11.4 g/dl on the 15th day, and between 12 and 14 g/dl after two months. Their symptoms had also resolved completely in two months. The patient with acute intoxication (5th case) had ingested copper oxychloride with suicidal intent. He was admitted with anuria and hemolytic anemia. After being hospitalized for fifteen days, he was diagnosed with chronic renal failure and was scheduled for a dialysis program. Acute poisoning is more deliberate, while chronic exposure may result in atypical findings. In conclusion, physicians working in primary care and EDs should consider copper poisoning in patients presenting with anemia, abdominal pain, headache, tightness in the chest, and leg and back pain.
Peritoneum has an intrinsic fibrinolytic activity that breaks the peritoneal adhesions. Peritoneal injuries with ischemia interfere this fibrinolytic activity and cause adhesions. Pentoxifylline, a methyl xanthine derivative, improves blood flow by decreasing its viscosity and also increases fibrinolytic activity in plasma. We hypothesized that pentoxifylline would increase peritoneal fibrinolysis and ameliorate adhesions. A rat model of peritoneal adhesion (cecal abrasion with gauze, n = 15 for each group) was used to test this hypothesis and cardinal parameters of peritoneal fibrinolysis were measured in peritoneal samples. No medication was given in control animals, while pentoxifylline was administered intraperitonealy (IP) (25 mg/kg, before abdominal closure to whole abdomen) or intravenously (IV) (25 mg/kg, for 9 days after operation) in the experimental groups. At postoperative day 10, peritoneal biopsies were obtained and adhesions were graded qualitatively. Activities and concentrations of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), tPA/PAI-1 complex and hydroxyproline contents were determined. Total adhesion scores were decreased in both treated groups. Mean levels of tPA concentration and tPA activity were increased in the treated groups compared to controls (p < 0.001 and p = 0.001, respectively). The tPA/PAI-1 complex levels were similar among the three groups. PAI-1 levels were lower in animals receiving IP pentoxifylline compared to control animals and those treated with IV pentoxifylline (p = 0.048, p = 0.015, respectively). Peritoneal hydroxyproline levels were similar among the three groups. Our results suggest that pentoxifylline administration either through IV or IP may reduce peritoneal adhesion formation probably by altering peritoneal fibrinolytic activity.
In adults, serum uric acid levels are positively correlated with body mass index (BMI) and hyperuricemia is considered to be a common lifestyle disorder related with obesity. However, the relation of serum uric acid levels with obesity has not been elucidated in children and adolescents. Serum uric acid levels were determined in 1,729 healthy children, consisted of 923 boys and 806 girls, aged 9.1 - 15.0 years. The incidence of hyperuricemia (defined as more than 7.0 mg/dl) in boys and girls were 8.8% and 0.6%, respectively. In 1,281 children out of all subjects, including 684 boys and 597 girls, height, weight, aspartate aminotransferase, and alanine aminotransferase were also determined and the correlations between serum uric acid levels and obesity were analyzed. BMI is popularly used as a standard indicator of obesity in adults. However, BMI increases without fat accumulation as children grow. In Japan, percentage of overweight (POW) is usually used as an alternative indicator for obesity. In general, children are evaluated as obesity, when POW is equal to or more than 20% (≥ 20%). Serum uric acid levels are positively correlated with obesity-related indicators, BMI and POW, in both boys and girls. Serum uric acid levels of the subjects with high POW (≥ 20%) are significantly higher than those of the subjects with low POW (< 20%) in both boys and girls. These results suggest that serum uric acid levels are significantly increased with obesity and could be used as one of obesity-related indicators even in early adolescence.
Niemann-Pick disease type C (NPC) is an inherited lipid storage disorder, characterized by a defect in intracellular trafficking of exogenous cholesterol that leads to the lysosomal accumulation of unesterified cholesterol. We report a Japanese patient with NPC caused by a homozygous c.2974 G > T mutation of the NPC1 gene, which predicts a glycine (GGG) to tryptophan (TGG) change at codon 992 (designated as p.G992W). This is a well-known NPC1 gene mutation that causes a unique phenotype of NPC, which has been limited to a single Acadian ancestor in Nova Scotia, Canada. Our patient characteristically started presenting with cataplexy at the age of 9 years. Recent studies have shown reduced hypocretin-1 levels in the cerebrospinal fluid (CSF) of narcoleptic patients with cataplexy. In our patient, the level of hypocretin-1 was determined as moderately low, 174 pg/ml (normal, > 200 pg/ml). To date, CSF levels of hypocretin-1 have been determined by using an identical assay method in 7 cases of NPC, including our case. All of the NPC cases with cataplexy demonstrated low levels of CSF hypocretin-1, confirming the association of reduced CSF hypocretin-1 levels with cataplexy in NPC.
Neuromyelitis optica (NMO) is clinically characterized by severe optic neuritis and transverse myelitis. In Japan, NMO has been named optic-spinal multiple sclerosis (OSMS) and it has been thought to be a subtype of multiple sclerosis (MS). However, several clinical and laboratory findings suggest NMO or OSMS is distinct from MS. Recently, the disease-specific antibody (NMO-IgG) was found in the serum from NMO patients, and its target antigen was identified as aquaporin-4 (AQP4) water channel protein which is mainly expressed in astroglial foot processes. However, the pathogenetic role of AQP4 in NMO remains unknown. We herein report a typical case of NMO in which immunohistochemical analysis showed a lack of AQP4 in the spinal cord lesions. The loss of AQP4 was evident in the central gray matter, especially in the perivascular lesions where immunoglobulins and complements were deposited, and glial fibrillary acidic protein (GFAP) staining was weak in those lesions. However, GFAP was strongly stained at the reactive astrogliosis surrounding the lesions. Myelin basic protein (MBP)-stained myelinated fibers were relatively preserved in the lesions where AQP4 was lost. In contrast to these NMO lesions, AQP4 was expressed predominantly in the gray matter in control spinal cords, and AQP4 was preserved in demyelinating MS lesions. Our findings suggest that astrocytic impairment associated with humoral immunity against AQP4 may be primarily involved in the lesion formation of NMO, and that the pathomechanisms of NMO are different from those of MS in which demyelination is the primary pathology.