Alternately binding probe competitive PCR as a simple, cost-effective, and accurate quantification method for JAK2V617F allele burden in myeloproliferative neoplasms

We developed a simple, cost-effective, and accurate JAK2 allele burden quantification method named alternately binding probe competitive PCR (ABC-PCR). ABC-PCR can be performed to quantify target JAK2 allele burdens in a single reaction. The throughput and running cost of ABC-PCR are markedly improved compared with those of allele-specific quantitative PCR (AS-qPCR). The quantification of samples with known JAK2 allele burdens revealed that ABC-PCR had a small assay-to-assay variation. The JAK2 allele burdens in the patients with myeloproliferative neoplasms measured by ABC-PCR and AS-qPCR showed a good fitting. ABC-PCR would be a powerful tool for quantifying target JAK2 allele burdens.     

Antibodies to myelin oligodendrocyte glycoprotein are not involved in the severity of chronic non-remitting experimental autoimmune encephalomyelitis

To elucidate the role of antibodies in development of chronic non-remitting experimental autoimmune encephalomyelitis (EAE) in C57BL/6 mice, which is a well-established Th1-mediated autoimmune disease, and the involvement of activation-induced cytidine deaminase (AID) in Th1-mediated function, we have investigated the myelin oligodendrocyte glycoprotein (MOG)-induced EAE in mice deficient of AID, which is absolutely required for class switching and somatic hypermutation. Following immunization with MOG, AID(-/-) had completely same levels of clinical and pathological severity of EAE when compared with AID(+/-) and AID(+/+), although AID(-/-) did not produce IgG and anti-MOG IgG. Similar levels of T cell proliferation and a modest increase of anti-MOG IgM synthesis were found in spleen cells of AID(-/-) stimulated with MOG. These results indicate that antibodies are not involved in development of EAE in C57BL/6 mice.     

Long-term outcome of ABO-incompatible living-donor liver transplantation: a single-center experience

Purpose  We report the long-term outcome of ABO-incompatible living donor liver transplantation (LDLT) performed in our hospital. Methods  We started the LDLT program in 1991 and from that year up to now (2008) 11 patients have received an ABO-incompatible graft. Results  Nine out of the 11 cases have survived from 3.7 years to 13.9 years (mean 7.3 years) and they are in good conditions at present. Seven patients were subjected to preoperative apheresis. Eight patients experienced acute rejection and of them, 6 experienced steroid-resistant rejection that was treated with deoxyspergualin and apheresis. One patient who suffered rapidly progressing rejection died due to liver failure. Three patients who were administered rituximab did not suffer severe rejection nor adverse effects. During the long-term follow up 5 recipients had major complications such as postoperative lymphoproliferative disease, post-transplantation diabetes mellitus, portal vein occlusion and biliary stenosis. But those complications were controlled under stable conditions. Conclusions  We concluded that long-term survival can be expected after ABO-incompatible LDLT provided perioperative complications such as humoral rejection are overcome.     

Oral Erythromycin Accelerates Impaired Gastrointestinal Motility After Endoscopic Mucosal Resection

Gastrointestinal motility may be impaired after endoscopic mucosal resection of gastric lesions. We investigated whether oral erythromycin could improve motility. Twenty patients were divided randomly into groups that received oral omeprazole with or without erythromycin. Motility was recorded overnight at 3 days before and 4 days after endoscopic resection using a microtransducer probe. In the group without erythromycin, gastric phase III activity decreased significantly after endoscopic resection, while it was increased significantly by erythromycin (P < 0.01). After resection, there were significantly more gastric phase III events in the erythromycin group (P < 0.05). The interval between the start of the evening meal and the initial gastric phase III activity was significantly prolonged after resection, while this interval was significantly shortened by erythromycin (P < 0.05). The gastric phase III cycle length was also significantly shortened by erythromycin (P < 0.05). Postprandial and fasting gastrointestinal motility were impaired after endoscopic resection, and postprandial as well as fasting motility were improved by oral erythromycin.     

Gasless laparoscopy-assisted distal gastrectomy is feasible and useful for non-obese patients with early gastric cancer

BACKGROUND/AIMS: To evaluate the feasibility and usefulness of gasless laparoscopy-assisted distal gastrectomy except when treating obese patients compared with open distal gastrectomy for early cancer. METHODOLOGY: We treated 92 patients with distal gastrectomy for early gastric cancer consecutively. Patients with massive submucosal invasion and/or LN swelling were allocated for the open method, and patients with slightly invasive submucosal cancer were allocated for gasless laparoscopy-assisted surgery. As exceptions we employed open surgery for overweight patients and gasless laparoscopy for elderly and/or feeble patients. RESULTS: We attempted to perform open and laparoscopy-assisted surgery on 52 and 40 patients, respectively. Three cases in the laparoscopy-assisted group were converted to open surgery because of obesity. The age was older and BMI was lower in the laparoscopy-assisted group. In terms of operative time and blood loss as well as postoperative recovery, the results for the laparoscopy-assisted group were superior to those of the open surgery group. There were no cases of cardiopulmonary complications for the laparoscopy-assisted group. CONCLUSIONS: Gasless laparoscopy-assisted distal gastrectomy is feasible and useful for early gastric cancer except when treating obese patients.     

A comparison of organ-tissue level body composition between college-age male athletes and nonathletes

The purpose of this study was to compare the characteristics of skeletal muscle (SM) mass and internal organ (liver and kidney) mass in resistance and/or high intensity trained collegiate athletes with nonathletes, and to examine the relationships between fat-free mass (FFM) and its major components of SM, liver mass, and kidney mass. Fifteen athletes and seventeen nonathletes volunteered for the study. FFM was measured by two-compartment densitometry. Contiguous magnetic resonance imaging was used to obtain images from the first cervical vertebrae to the ankle joint (no inter-slice gap) for each subject, and SM, liver and kidney cross-sectional areas and organ-tissue volumes were determined. Organ-tissue volumes (cm (3)) were converted to mass (kg) by multiplying the volumes by the assumed constant density of the tissues. On average, athletes had greater FFM (69.1 kg) than the nonathletes (52.6 kg). SM, liver, and kidney masses in athletes (33.0 kg, 1.84 kg and 0.39 kg, respectively) were higher compared with nonathletes (23.5 kg, 1.39 kg and 0.31 kg, respectively). When the various determinants of FFM were expressed as ratios, it was determined that the ratio of SM mass to FFM was higher in athletes (47.7 %) than nonathletes (44.7 %), and the ratios of liver and kidney to FFM were similar for the two groups (2.6 % and 0.6 %, respectively, for athletes; and 2.7 % and 0.6 %, respectively, for nonathletes). A strong correlation between FFM and SM mass was observed in athletes and nonathletes and the slopes of these regression lines were almost identical and parallel. FFM was also significantly correlated with liver and kidney mass for both athletes and nonathletes. This study suggests that SM, liver, and kidney masses are increased by FFM accumulation in resistance and/or high intensity trained athletes.     

Usefulness of beta-D glucan in diagnosing Pneumocystis carinii pneumonia and monitoring its treatment in a living-donor liver-transplant recipient

Pneumocystis carinii pneumonia (PCP) is one of the fatal complications encountered after liver transplantation. The diagnosis of PCP is sometimes very difficult, because detection of the bacteria itself is not easy under some conditions, and the serum level of the chemical mediator is not yet considered to be a definitive diagnostic marker. We report a case of PCP that occurred 3 months after transplantation in a living-donor liver-transplant recipient; the disease developed during the course of outpatient follow-up when the patient's condition was stable. The patient was maintained with the usual level of immunosuppressants, using tacrolimus, steroid, and mycophenolate mofetil. The patient had a dry cough with mild fever, and a chest computed tomography (CT) scan showed a reticular shadow in the left lung field. The plasma level of beta-d glucan was high (135 pg/ml). We suspected an invasive fungal infection, but no pathogen was detected by routine fungal culture and cytology. Finally, P. carinii was detected by polymerase chain reaction (PCR), and we started treatment with trimethoprim-sulfamethoxazole (TMP/SMX) combined with an antifungal agent. During this period, the level of beta-d glucan correlated with the patient's clinical symptoms; this marker was very useful for monitoring the treatment of PCP in this living-donor liver-transplant recipient.     

Hydrogen sulfide inhibits activity of three isoforms of recombinant nitric oxide synthase

To clarify the presence of cross-talk between H(2)S and NO, we investigated effect of NaHS, an H(2)S donor, on activity of recombinant NO synthase (NOS) isoforms. Activity of all nNOS, iNOS and eNOS was inhibited by NaHS (IC(50): 0.13-0.21 mM). In contrast, Na(2)SO(3), L-cysteine and threo-1,4-dimercapto-2,3-butanediol, a reductant, exerted poor inhibition of NOS activity. Increasing concentrations of tetrahydrobiopterin (BH(4)) reversed the NaHS inhibition of nNOS and eNOS, but not iNOS. Our data thus demonstrate inhibition of three NOS isoforms by NaHS/H(2)S, and suggest involvement of interaction of NaHS/H(2)S with BH(4) in inhibition of nNOS and eNOS, but not iNOS.