Differential Reactivation of Cytomegalovirus and Epstein-Barr Virus in Patients with B Cell Lymphoma.

Although cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are considered latent viruses, their reactivation occurs in immunosuppressed conditions. We previously reported that CMV and EBV are reactivated in patients receiving immunosuppressive therapy and/or chemotherapy. This retrospective, single-center study aimed to determine the frequency of viral reactivation and clinical characteristics of patients with B cell lymphoma (B-ML) receiving chemotherapy. Twenty-four patients (mean age 73 years, range 40-87 years; male-to-female ratio, 15:9) with diffuse large B cell lymphoma (n = 15), follicular lymphoma (n = 8), or mantle cell lymphoma (n = 1) were enrolled. Serum CMV and EBV DNA levels were analyzed using quantitative real-time polymerase chain reaction in patients with B-ML receiving chemotherapy. We determined the cumulative reactivation of each virus and analyzed the relationship between viral reactivation and clinical characteristics. Three patients experienced relapse or refractory (R/R) disease and the others had de novo lymphomas. The frequencies of CMV and EBV reactivations were 54.2% and 37.5%, respectively. CMV reactivation occurred significantly earlier during chemotherapy courses in R/R patients than in de novo patients (p = 0.0038), while EBV reactivation was frequently found before treatment. Baseline serum levels of soluble interleukin-2 receptor were higher (4318.0 vs. 981.1 U/mL, p = 0.010) and hemoglobin levels were lower (11.1 vs. 13.0 g/dL, p = 0.0038) in patients with EBV reactivation than in those without reactivation. These findings were not observed in patients with CMV reactivation. CMV reactivation was associated with iatrogenic immunosuppression, whereas EBV reactivation was related to immunosuppression by lymphoma, indicating that the mechanisms of these viral reactivations differed.

Nephrotic Syndrome without Hematuria due to Infection-Related Glomerulonephritis Mimicking Minimal-Change Disease in a Child.

Nephrotic syndrome without hematuria due to infection-related glomerulonephritis is uncommon. The present report describes a case of nephrotic syndrome due to infection-related glomerulonephritis without hematuria and hypertension in an older child. A 14-year-old boy was referred to our hospital because of a 5-day history of fever, nausea, weight gain and recent leg edema without hypertension. Laboratory data showed nephrotic-range proteinuria, hypoalbuminemia, mild hypocomplementemia and acute renal injury without hematuria. Although, due to the clinical presentation, minimal-change nephrotic syndrome was mostly suspected, a renal biopsy showed endocapillary hypercellularity mainly of mononuclear cells with segmental mesangiolytic changes. Fine granular IgG and C3 deposits were noted by an immunofluorescent study; many relatively small electron-dense deposits were observed electron-microscopically. These findings led to the diagnosis of nephrotic syndrome due to infection-related endocapillary proliferative glomerulonephritis, although the causative organism of his nephritis was not detected. He recovered with rest and dietary cure. When we examine an acute nephrotic child, infection-related glomerulonephritis should be considered as the differential diagnosis to avoid unnecessary use of corticosteroids.

Temporal coding of insulin action through multiplexing of the AKT pathway.

One of the unique characteristics of cellular signaling pathways is that a common signaling pathway can selectively regulate multiple cellular functions of a hormone; however, this selective downstream control through a common signaling pathway is poorly understood. Here we show that the insulin-dependent AKT pathway uses temporal patterns multiplexing for selective regulation of downstream molecules. Pulse and sustained insulin stimulations were simultaneously encoded into transient and sustained AKT phosphorylation, respectively. The downstream molecules, including ribosomal protein S6 kinase (S6K), glucose-6-phosphatase (G6Pase), and glycogen synthase kinase-3ß (GSK3ß) selectively decoded transient, sustained, and both transient and sustained AKT phosphorylation, respectively. Selective downstream decoding is mediated by the molecules' network structures and kinetics. Our results demonstrate that the AKT pathway can multiplex distinct patterns of blood insulin, such as pulse-like additional and sustained-like basal secretions, and the downstream molecules selectively decode secretion patterns of insulin.

Latent process genes for cell differentiation are common decoders of neurite extension length.

A latent process involving signal transduction and gene expression is needed as a preparation step for cellular function. We previously found that nerve growth factor (NGF)-induced cell differentiation has a latent process, which is dependent on ERK activity and gene expression and required for subsequent neurite extension. A latent process can be considered as a preparation step that decodes extracellular stimulus information into cellular functions; however, molecular mechanisms of this process remain unknown. We identified Metrnl, Dclk1 and Serpinb1a as genes that are induced during the latent process (LP) with distinct temporal expression profiles and are required for subsequent neurite extension in PC12 cells. The LP genes showed distinct dependency on the duration of ERK activity, and they were also induced during the latent process of PACAP- and forskolin-induced cell differentiation. Regardless of neurotrophic factors, expression levels of the LP genes during the latent process (0-12 hours), but not phosphorylation levels of ERK, always correlated with subsequent neurite extension length (12-24 hours). Overexpression of all LP genes together, but not of each gene separately, enhanced NGF-induced neurite extension. The LP gene products showed distinct spatial localization. Thus, the LP genes appear to be the common decoders for neurite extension length regardless of neurotrophic factors, and they might function in distinct temporal and spatial manners during the latent process. Our findings provide molecular insight into the physiological meaning of the latent process as the preparation step for decoding information for future phenotypic change.

Determination of cyanide in blood by electrospray ionization tandem mass spectrometry after direct injection of dicyanogold.

An electrospray ionization tandem mass spectrometric (ESI-MS-MS) method has been developed for the determination of cyanide (CN(-)) in blood. Five microliters of blood was hemolyzed with 50 µL of water, then 5 µL of 1 M tetramethylammonium hydroxide solution was added to raise the pH of the hemolysate and to liberate CN(-) from methemoglobin. CN(-) was then reacted with NaAuCl(4) to produce dicyanogold, Au(CN)(2)(-), that was extracted with 75 µL of methyl isobutyl ketone. Ten microliters of the extract was injected directly into an ESI-MS-MS instrument and quantification of CN(-) was performed by selected reaction monitoring of the product ion CN(-) at m/z 26, derived from the precursor ion Au(CN)(2)(-) at m/z 249. CN(-) could be measured in the quantification range of 2.60 to 260 µg/L with the limit of detection at 0.56 µg/L in blood. This method was applied to the analysis of clinical samples and the concentrations of CN(-) in the blood were as follows: 7.13 ± 2.41 µg/L for six healthy non-smokers, 3.08 ± 1.12 µg/L for six CO gas victims, 730 ± 867 µg for 21 house fire victims, and 3,030 ± 97 µg/L for a victim who ingested NaCN. The increase of CN(-) in the blood of a victim who ingested NaN(3) was confirmed using MS-MS for the first time, and the concentrations of CN(-) in the blood, gastric content and urine were 78.5 ± 5.5, 11.8 ± 0.5, and 11.4 ± 0.8 µg/L, respectively.

The NMDA receptor NR2A subunit regulates proliferation of MKN45 human gastric cancer cells.

The present study investigated proliferation of MKN28 and MKN45 human gastric cancer cells regulated by the N-methyl-d-aspartate (NMDA) receptor subunit. The NMDA receptor antagonist dl-2-amino-5-phosphonovaleric acid (AP5) inhibited proliferation of MKN45 cells, but not MKN28 cells. Of the NMDA subunits such as NR1, NR2 (2A, 2B, 2C, and 2D), and NR3 (3A and 3B), all the NMDA subunit mRNAs except for the NR2B subunit mRNA were expressed in both MKN28 and MKN45 cells. MKN45 cells were characterized by higher expression of the NR2A subunit mRNA and lower expression of the NR1 subunit mRNA, but MKN28 otherwise by higher expression of the NR1 subunit mRNA and lower expression of the NR2A subunit mRNA. MKN45 cell proliferation was also inhibited by silencing the NR2A subunit-targeted gene. For MKN45 cells, AP5 or knocking-down the NR2A subunit increased the proportion of cells in the G(1) phase of cell cycling and decreased the proportion in the S/G(2) phase. The results of the present study, thus, suggest that blockage of NMDA receptors including the NR2A subunit suppresses MKN45 cell proliferation due to cell cycle arrest at the G(1) phase; in other words, the NR2A subunit promotes MKN45 cell proliferation by accelerating cell cycling.

[Perioperative management of airway foreign bodies in 35 pediatric patients].

We retrospectively reviewed 33 patients (35 cases) who underwent foreign body removal at our institution from 1995 through 2003. Male-female ratio was 21 : 12 and the most frequent age was one year. The most common foreign bodies (FBs) were nuts (n = 14) and plastics (n = 7). A repeater (3 cases) had mental retardation. Patients were referred to our institution with an average interval of 90 hours, and after 1.3 hospitals. All the patients were managed with general anesthesia. Direct laryngoscopy was performed to extract FBs in 11 cases suspected of having pharyngeal or laryngeal FBs. In 24 cases suspected of having tracheobronchial FBs, the trachea was intubated and a flexible fiberoptic bronchoscopy was performed to locate the FBs. FBs were found in the trachea in 2 cases and in the bronchus in 18 patients and were successfully extracted by rigid bronchoscopy in 10 cases. All the patients were admitted for fear of laryngotracheal edema. There were no significant postoperative complications.

Beneficial effect of intracellular free high-mannose oligosaccharides on cryopreservation of mammalian cells and proteins.

The cryoprotective effect of intracellular free high-mannose oligosaccharides (HMOS) on mammalian cells and proteins was examined by monitoring PC-12 cell viability and assaying protein kinase C (PKC)-epsilon activity. 1-Deoxymannojirimycin, an inhibitor of alpha-mannosidase, to cause an increase in intracellular free HMOS, significantly rescued PC-12 cells with 2-h freezing insult at -15 degrees C in a concentration (1-50mM)- and pretreatment time (48-72h)-dependent manner, as compared with unpretreated cells; full rescue from freezing injury was obtained with 1-deoxymannojirimycin at more than 25mM for 48-h pretreatment and more than 3mM for 72- and 96-h pretreatment. For PC-12 cells pretreated with 1-deoxymannojirimycin at 1mM for 72h, thawed cell viability after more than 8-w cryopreservation at -80 degrees C in 10% (v/v) dimethyl sulfoxide was much higher than that for cells without pretreatment. PKC-epsilon activity was well preserved after 16-h cryopreservation at -20 degrees C in the presence of mannose 9-N-acetylglucosamine 2 (Man9-GlcNAc2) (1 mM), an HMOS, while the activity was reduced to 15% without Man9-GlcNAc2. Collectively, the results of the present study suggest that intracellular free HMOS is a key molecule to protect mammalian cells and proteins from freezing injury; in other words, HMOS could be a new target for cryopreservation of mammalian cells and proteins.

[Perioperative management for laparoscopic repair of congenital duodenal atresia].

BACKGROUND: Laparoscopic surgery has been applied even to neonates. To examine the safety and efficacy of laparoscopic repair of neonatal congenital duodenal atresia, we compared the 5 laparoscopic cases with the 5 conventional surgical cases. METHODS: The charts were retrospectively reviewed to investigate the anesthetic management, perioperative status and complications in the most recent 5 cases each of laparoscopic and conventional surgeries. RESULTS: There was a tendency to avoid laparoscopic repair in the patients with congenital heart disease. There were no intraoperative complications in both groups. Laparoscopic group exhibited less blood loss but longer operation time. In the two out of five laparoscopic cases re-operation was required due to technical issues, and the group needed a longer period before starting enteral feeding and longer hospitalization. CONCLUSIONS: Up to this time, laparoscopic repair of congenital duodenal atresia exhibited few advantages over conventional open repair.

[Anesthetic management of a 1.7-kg premature infant undergoing thoracoscopic thoracic duct ligation].

We report successful anesthetic management of a 1.7-kg premature infant who underwent thoracoscopic thoracic duct ligation under general anesthesia. She was born at 30 weeks gestation with birth weight of 1,546 g and was suffering from respiratory distress due to persistent right chylothorax for two months after birth. Chest tube drainage, fasting and intrapleural fibrin glue did not reduce her right chylothorax. Thoracoscopic thoracic duct ligation was scheduled on her day 64 under general anesthesia. The tracheal tube end was placed in the midtrachea and carbon dioxide was insufflated into the operative side of the thorax. During thoracoscopy her left lung was ventilated with the right lung pressed with spatulaes, but her respiratory status did not deteriorate so much despite of unilateral ventilation. We speculate that, due to massive right chylothorax, her pulmonary blood flow had already shifted to the left lung, therefore intraoperative substantial left unilateral lung ventilation exerted minimal effect on her respiratory status. The operation was successful and she was weaned from the ventilator on the following day.

A case of diffuse panbronchiolitis (DPB) in a patient positive for HTLV-1.

Diffuse panbronchiolitis (DPB), an important cause of progressive obstructive lung disease in the Far East, represents a distinctive sinobronchial syndrome with typical radiologic and histologic features. Human T-cell lymphotrophic virus (HTLV-1) is a retrovirus that clinically and experimentally suppresses T-cell function and immune responses. The clinical and immunologic features of DPB in HTLV-1 carriers are unclear, because DPB and HTLV-1 endemic areas around the world are mostly non-overlapping. However, both diseases are endemic in Japan. We present a patient with DPB positive for HTLV-1 whose cellular and humoral immune responses were markedly impaired. Six y after diagnosis of DPB, the patient developed respiratory failure and died in spite of treatment with clarithromycin.