Highly sensitive detection of Epstein-Barr virus-infected cells by EBER flow FISH.

When Epstein-Barr virus (EBV) infection is suspected, identification of infected cells is important to understand the pathogenesis, determinine the treatment strategy, and predict the prognosis. We used the PrimeFlow™ RNA Assay Kit with a probe to detect EBV-encoded small RNAs (EBERs) and multiple surface markers, to identify EBV-infected cells by flow cytometry. We analyzed a total of 24 patients [11 with chronic active EBV disease (CAEBV), 3 with hydroa vacciniforme lymphoproliferative disorder, 2 with X-linked lymphoproliferative disease type 1 (XLP1), 2 with EBV-associated hemophagocytic lymphohistiocytosis, and 6 with posttransplant lymphoproliferative disorder (PTLD)]. We compared infected cells using conventional quantitative PCR methods and confirmed that infected cell types were identical in most patients. Patients with CAEBV had widespread infection in T and NK cells, but a small amount of B cells were also infected, and infection in patients with XLP1 and PTLD was not limited to B cells. EBV-associated diseases are believed to be complex pathologies caused by EBV infecting a variety of cells other than B cells. We also demonstrated that infected cells were positive for HLA-DR in patients with CAEBV. EBER flow FISH can identify EBV-infected cells with high sensitivity and is useful for elucidating the pathogenesis.

Pediatric liver failure with massive sinusoidal infiltration of histiocytes.

Histiocytic neoplasms, such as Langerhans cell histiocytosis (LCH) and disseminated juvenile xanthogranuloma (JXG), can involve the liver and sometimes cause liver failure. We aimed to classify non-LCH histiocytic proliferating disorders that do not exhibit typical disseminated JXG histology. We examined four pediatric patients who presented with liver failure and splenomegaly. Two patients with liver cirrhosis without cholestasis underwent liver transplantation (LT). The other two patients presented with giant cell hepatitis causing neonatal/infantile acute liver failure (ALF). The infantile ALF patient also underwent LT. Liver dysfunction developed after LT in all three transplant cases and the grafts exhibited massive sinusoidal infiltration of histiocytes with hemophagocytosis, similar to the native liver. The neonatal ALF patient was treated with an LCH-type chemotherapy regimen, and is alive and well at 18 months. Infiltrating histiocytes were positive for CD68 and CD163, and negative for CD1a, CD207, and S-100 protein. The BRAF V600E mutation was not present. Liver histological findings were not consistent with conventional disseminated JXG or LCH, although the histological findings in other organs overlapped those of well-known histiocytic neoplasms. The histological and immunohistochemical findings of infiltrating histiocytes suggest that these four cases constituted a disseminated JXG-like systemic disease.

Small-Cell Lung Cancer Comorbid with Pulmonary Mycobacterium avium Infection: A Case Report.

BACKGROUND: Small-cell lung cancer (SCLC) rarely coexists with pulmonary Mycobacterium avium intracellular complex (MAC) infection. The key drug for SCLC treatment is etoposide, which is metabolized by cytochrome P-450 (CYP) 3A4. Meanwhile, the key drugs for pulmonary MAC infection are clarithromycin (CAM) and rifampicin (RFP), and their metabolism influences CYP3A4. Therefore, treatment of concurrent SCLC and pulmonary MAC infection is difficult, and to the best of our knowledge, no report of treatments for concurrent SCLC and pulmonary MAC infection has been published. Patient Concerns and Diagnoses: A 65-year-old man presented to our hospital with abnormal findings of chest computed tomography: (1) a hilar region nodule in the left lung and mediastinal lymphadenopathy and (2) a thick-walled cavity lesion in the right upper lobe of the lung. After further examinations, the former lesions were diagnosed as SCLC, cT4N3M0, stage IIIC and the latter as pulmonary MAC infection, fibrocavitary disease. INTERVENTIONS AND OUTCOMES: Concurrent treatment was conducted with discontinuation of CAM and RFP before and after etoposide administration. Specifically, intravenous cisplatin and etoposide were administered on day 1 and days 1-3, respectively, and CAM, RFP, and ethambutol (EB) were administered orally on days 6-22 every 4 weeks. Concurrent radiotherapy was added to the drug administration on days 1-27 of the first cycle. The chemotherapy was continued for 4 cycles, followed by continuation of CAM and RFP administration. EB was discontinued because of optic nerve disorder. The treatments were conducted completely and safely, and both of the SCLC lesions and the MAC lesion were improved. CONCLUSIONS: Treatments for concurrent SCLC and pulmonary MAC infection may be successfully conducted with discontinuation of CAM and RFP before and after etoposide administration.

Acute renal failure likely due to acute nephritic syndrome associated with typhoid fever.

A 45-year-old woman was admitted to our hospital because of high fever and malaise. She also presented abnormal urine findings including macrohematuria and proteinuria with transient renal insufficiency. Salmonella Typhi was isolated from her blood and stool culture, and then she was diagnosed as having typhoid fever. Salmonella enterica serotype typhi infection can be complicated by renal involvement, although rare. In Japan, few cases have been reported about acute nephritic syndrome in typhoid fever. Here, we report a case of endemic acquired typhoid fever associated with acute renal failure probably due to acute nephritic syndrome successfully treated with levofloxacin.

Simultaneous operation of ischemic heart disease, abdominal aortic aneurysm, and rectal cancer.

A 68-year-old man with ischemic heart disease, abdominal aortic aneurysm, and rectal cancer was referred. Coronary angiography indicated triple-vessel disease with jeopardized collaterals, and dipyridamole myocardial scintigraphy disclosed no viability in the inferior, posterior, and lateral walls. Abdominal computed tomography scanning revealed an infrarenal abdominal aortic aneurysm, 65 mm in diameter, with an expanding rate of 8 mm/year. Barium enema revealed stenosis 4 cm in length 5 cm inward from the anal verge, and an endoscopic finding was ulcerated type tumor with a clear margin and circumferential stenosis. Histological examination of a biopsy specimen revealed adenocarcinoma, and the clinical stage in the Japanese classification of colorectal carcinoma was II according to other examinations. Simultaneous operations were scheduled because of the jeopardized collaterals of the coronary arteries, rapid expansion of the aneurysm, and subileus due to the cancer. The patient underwent simultaneous off-pump coronary artery bypass grafting to the left anterior descending artery with the in situ internal thoracic artery through a median sternotomy, abdominal aortic aneurysm repair with a tube graft through a median laparotomy, and the Miles' operation with total mesorectal excision. Although infection of the perineal wound was postoperatively recognized, it remained local and was healed with irrigation only. The patient is doing well 12 months after the operation, without myocardial ischemic symptoms or recurrence of the cancer.

Liddle's syndrome caused by a novel mutation in the proline-rich PY motif of the epithelial sodium channel beta-subunit.

Liddle's syndrome is an autosomal dominant form of salt-sensitive hypertension and has been shown to be caused by missense or frameshift mutations in the amiloride-sensitive epithelial sodium channel (ENaC), which is composed of three subunits: alpha, beta, and gamma. All disease mutations either remove or alter amino acids of the target proline-rich PPPxY sequence (PY motif) of beta- or gamma-ENaC and result in increased channel activity. In this report, we present a family with Liddle's syndrome whose abnormality is caused by a novel missense mutation, P616R, in the PY motif of the betaENaC. Functional studies using the P616R mutant expressed in Xenopus oocytes showed an approximately 6-fold increase in the amiloride-sensitive sodium channel activity compared with that of the wild type. These findings provide additional clinical evidence that a conserved PY motif is critically important for the regulation of ENaC activity.

Primary peritonitis associated with streptococcal toxic shock-like syndrome: report of a case.

Several reports over the past 15 years describe severe group A streptococcal infections causing septic shock, soft-tissue necrosis, and multiple organ failure; a phenomenon known as streptococcal toxic shock-like syndrome (TSLS). However, primary peritonitis associated with TSLS is rare. We report the case of a 40-year-old man admitted with pain in both thighs, hypotension, and severe abdominal pain. His daughter had been diagnosed with streptococcal pharyngitis 3 days earlier. We performed an emergency laparotomy for peritonitis, and culture of the ascites was positive for group A beta -hemolytic streptococcus (GAS). Further serotyping of the isolated GAS strain revealed the T-type 22 and the pyrogenic exotoxin gene, spe-C. The criteria for TSLS were clearly met, including the isolation of GAS from ascites, hypotension, liver failure, renal failure, coagulopathy, myositis, and a generalized erythematous macular rash with desquamation.

Possible impairment of transcardiac utilization of adiponectin in patients with type 2 diabetes.

OBJECTIVE: Adiponectin, an adipocyte-derived protein, has been suggested to enhance insulin sensitivity and prevent atherosclerosis. Circulating adiponecin levels are reduced in states of insulin resistance such as type 2 diabetes. We examined transcardiac utilization of adiponectin in patients with and without type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 17 male type 2 diabetic patients and 17 male nondiabetic patients were investigated. Venous blood samples were taken to measure glucose and lipid variables. Blood samples for the measurement of adiponectin were collected simultaneously from the aortic root and coronary sinus. Angiographic semiquantitative stenosis score of coronary artery was also evaluated. RESULTS: The adiponectin levels in both the aortic root and coronary sinus in the diabetic patients were significantly lower than those in the nondiabetic patients. The adiponectin level was significantly lower in the coronary sinus than in the aortic root in the nondiabetic patients, but there was no significant difference between adiponectin levels in the aortic root and coronary sinus in the diabetic patients. The total stenosis score, as an index of severity of coronary artery stenosis, was significantly higher in the diabetic patients than in the nondiabetic patients. The stenosis score was correlated with the degree of transcardiac utilization of adiponectin from the aortic root to coronary sinus in the nondiabetic patients but not in the diabetic patients. CONCLUSIONS: Diabetic patients not only have a decreased adiponectin level in the basal state compared with nondiabetic patients but also have impaired utilization of adiponectin in the coronary artery and/or the heart, which may promote the development of atherosclerosis.

Experimental studies to estimate the intestinal viability in a rat strangulated ileus model using a dielectric parameter.

This study aimed at establishment of adequate evaluation of intestinal viability before release of the strangulated intestine by measuring electrical properties. In rats a 20-cm segment of the distal ileum and mesentery was strangulated. The intestine was strangulated for 0, 2, 15, 45, 90, and 120 min. The conductance and capacitance obtained by impedance analyzer were used to calculate tan(delta)m in the strangulated intestine. ATP was measured as well. In another experiment, after various periods of strangulation, tan(delta)m was measured just before release of the strangulation. Rats were divided into Group A (survived for 7 days) and Group D (died within 7 days). There was a positive correlation between tan(delta)m and ATP levels (P < 0.01). And tan(delta)m was significantly greater in Group A than in Group D (P < 0.05). There were no deaths at a tan(delta)m value of 2.36 or more and no survivors at a tan(delta)m of less than 2.20 except for one rat. These results suggested that tan(delta)m may be a useful index of the viability of the strangulated intestine before reperfusion.

Translocation of Salmonella typhimurium in rats on total parenteral nutrition correlates with changes in intestinal morphology and mucus gel.

OBJECTIVE: We tested whether alterations in intestinal morphology and mucus gel correlate with differences in Salmonella typhimurium translocation between rats treated with total parenteral nutrition (TPN) and rats given a diet of chow. METHODS: Twenty-seven male Wistar rats were assigned to one of two groups: one received TPN for 14 d and the other (control) received standard rat chow and water ad libitum. Salmonella typhimurium (5 x 10(8) cells; GIFU 12142) was injected into a closed ileal loop. Portal venous blood (PVB), inferior vena cava blood (IVCB), and mesenteric lymph nodes (MLNs) were sampled for evaluation of bacterial translocation. Sections of the loop were prepared and stained with hematoxylin and eosin, periodic acid-Schiff (PAS), and fluorescein isothiocyanate-labeled Ulex europaeus agglutinin I (FITC-UEA-I) for image analysis. Perimeter, mucosal thickness, villus area, and positively stained mucus area were measured. A fluorescent antibody study was also done. RESULTS: Organisms were found in cultures of 1 in 13 control rats and 9 in 14 TPN rats. There were more bacteria in MLNs than in PVB or IVCB. There was no increase in the number of bacteria over time in PVB, IVCB, or MLNs. Perimeter and villus area (P < 0.001) and mucosal thickness (P < 0.01) were significantly smaller in the TPN group than in the control group. The positively stained mucus area was significantly smaller in the TPN group than in the control group (P < 0.05 with PAS, P < 0.01 with FITC-UEA-I). Salmonella typhimurium invaded specifically through Peyer's patches. In all culture-negative samples, bacteria were trapped by the mucous layer, with a very small number attached to the epithelial surface. CONCLUSION: Significant villous atrophy and reduction of mucus play an important role in the rapid translocation of S. typhimurium through Peyer's patches in rats after 2 wk of TPN.

Specific CTL activity of CD8+ TCR Vbeta14+ T cell in mouse 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis.

We analyzed the functional role of CD8+ T-cell receptor (TCR) Vbeta14+ T cells, which increased specifically in the lamina propria in 2,4,6-trinitrobenzene sulfonic acid (TNBS) -induced colitis. Cytotoxic activity and cytokine production in CD8+ TCR Vbeta14+ T-cell clones were analyzed by 51Cr release assay and enzyme-linked immunosorbent assay, respectively. Cell transfer studies using these clones were performed. Established T-cell clones showed specific cytotoxic activity against TNBS-conjugated self spleen cells, and this cytotoxicity was completely inhibited by anti-TCR Vbeta14 monoclonal antibody. These clones produced interferon (IFN) - gamma in their culture supernatant, but neither interleukin (IL) - 2 nor IL-4. Histological findings of the colon in mice, which received clone transfer after enema with suboptimal doses of TNBS, showed massive colitis. Our results indicate that CD8+ TCR Vbeta14+ T cells had a cytotoxic T-lymphocyte function induced by Th-1 T-cell response and played a pathogenic role in the development of TNBS-induced colitis.

Dielectric spectrogram for evaluating ischemic microstructural changes of the liver in simple cold preservation.

PURPOSE: Monitoring ischemic liver injury is important in liver transplantation. We previously reported that dielectric properties were correlated with tissue adenosine triphosphate during liver preservation; however, it is unknown what the changes in dielectric properties are based on. Using rats, we studied the relationship between dielectric parameters and microstructural changes in liver tissues during preservation. METHODS: We serially measured the percent decrease in conductivity and the percent increase in relative permittivity in rat livers, and determined the areas of cellular and extracellular components using a charge-coupled device microscope and an NIH image. Thereafter, we evaluated the relationship between dielectric parameters and microstructural changes in the liver tissues. RESULTS: A positive linear and exponential correlation was found between the percent decrease in conductivity and the percent decrease in extracellular components ( P < 0.01). A positive linear and exponential correlation was also found between the percent increase in relative permittivity and the percent increase in cellular components ( P < 0.01). CONCLUSIONS: Our results suggest that the serial changes in tissue conductivity and the relative permittivity reflect changes in liver tissue microstructures during cold preservation.