CMV DNA detection in dried blood spots for diagnosing congenital CMV infection in Japan

Human cytomegalovirus (CMV) is a leading congenital infectious agent in developed countries. In the past, the incidence of congenital infection has been rather low in Japan because a high seroprevalence of CMV present in young women. However, this seroprevalence has been decreasing in recent years, so that the incidence of congenital CMV infection in Japanese neonates may increase and approach the level seen in other developed countries. The method was used for detecting CMV DNA reported by Barbi et al. [Barbi et al. (1996): Clin Diagn Virol 6:27-32] using a dried blood spot on filter paper, to diagnose congenital CMV infection in Japanese neonates. This method is effective and less laborious than virus isolation both for epidemiological studies and for identifying asymptomatic infected babies. Japanese neonates (1,176) were examined; two of who were asymptomatic were found to be infected.     

Granulysin produced by uterine natural killer cells induces apoptosis of extravillous trophoblasts in spontaneous abortion

Immune changes are known to occur in recurrent spontaneous abortion, but it is unclear whether either maternal natural killer (NK) cells or T cells attack fetus-derived trophoblasts. To clarify the immunological causes of spontaneous abortion, we examined the relationship between cytotoxic granule proteins in decidual lymphocytes, such as granulysin, granzyme B, and perforin, and the induction of apoptosis in extravillous trophoblasts (EVTs). The number of granulysin-positive CD56^bright NK cells increased significantly in the decidua basalis during spontaneous abortion compared with normal pregnancy; however, granzyme B- and perforin-positive cells did not change. Interestingly, the expression of granulysin was also detected in the nuclei of EVTs in spontaneous abortion samples. When IL-2-stimulated CD56^bright NK cells were cocultured with EVT cells (HTR-8/SV40neo), granulysin was found initially in the cytoplasm and then accumulated in the nuclei of the HTR-8/SV40neo cells. Furthermore, transfected cells expressing a GFP-granulysin fusion protein induced apoptosis in HTR-8/SV40neo cells independently of caspases. Our results suggest that granulysin-positive uterine NK cells attack EVTs; subsequently, the uNK-derived granulysin actively accumulates in the nuclei of EVTs, causing the death of EVTs due to apoptosis. These data support a new apoptosis pathway for trophoblasts via uNK-derived granulysin, suggesting that granulysin is involved in spontaneous abortion.     

Visualization of lymphatic basin from the tumor using magnetic resonance lymphography with superparamagnetic iron oxide in patients with thoracic esophageal cancer

OBJECTIVE: To evaluate magnetic resonance (MR) lymphography with submucosal injection of superparamagnetic iron oxide (SPIO) for imaging lymphatic pathways from thoracic esophageal cancer. METHODS: In 24 patients with esophageal cancer, SPIO was injected into the submucosal layer of the peritumoral region endoscopically and MR lymphography was conducted. In study 1, fast spoiled gradient-recalled acquisition using a steady-state (FSPGR) sequence was performed from the neck to the upper abdomen before and at 20, 40, and 60 minutes after injection in 10 patients. In study 2, FSPGR and spin echo T1-weighted images were obtained after injection in 14 patients. Areas scanned were the neck to the upper mediastinum and the upper abdomen. RESULTS: In study 1, at 20 minutes after injection, the signal of each lymph node appeared attenuated when compared with precontrast images. The signal-to-noise ratio in lymph nodes exhibiting influx of SPIO was significantly lower than that found on precontrast images (P < 0.0005). In study 2, influx to the neck lymph nodes was detected in 8 patients (64.3%), whereas influx to the upper abdominal lymph nodes was detected in 13 (92.9%). CONCLUSIONS: Magnetic resonance lymphography with SPIO could visualize the lymphatic pathways draining from the injection site and the location of lymph nodes exhibiting influx of SPIO in patients with thoracic esophageal cancer.     

The presence of BRAF point mutation in adult papillary thyroid carcinomas from atomic bomb survivors correlates with radiation dose

In papillary thyroid carcinogenesis, the constitutively activated mitogen-activated protein (MAP) kinase signaling pathway caused by a genetic alteration such as RET/PTC rearrangement or mutation of RAS and BRAF genes, is thought to be a major early event. Among these, the recently identified BRAF(V600E) mutation has been found at high frequency in adult patients with papillary thyroid carcinoma (PTC). However, the association between this mutation and radiation exposure in adult PTC is still unknown. In this study, we examined the BRAF(V600E) mutation in 64 PTCs among adult atomic bomb survivors in Hiroshima, Japan, comprising 17 nonexposed (0 mGy) and 47 exposed patients who developed the carcinoma after the bombing, and assessed the association of BRAF(V600E) mutation with clinico-pathological and epidemiological variables. The median radiation dose in PTCs with the BRAF(V600E) mutation was significantly lower than that without the mutation (18.5 vs.156.9 mGy, Wilcoxon rank-sum test, P=0.022). A significant difference was found in the median latency period (years elapsed from atomic bombing to diagnosis) between exposed patients with and without BRAF(V600E) mutation (29 vs. 21 yr, Wilcoxon rank-sum test, P=0.014). These findings were further confirmed by logistic regression analysis with BRAF(V600E) mutation status as a dependent variable and taking into account possible interactions between the variables. We found that the log-transformed radiation dose and latency period were independently associated with the BRAF(V600E) mutation (P=0.039 and P=0.010, respectively). These results suggest that involvement of BRAF mutation in thyroid carcinogenesis in exposed people may differ from that in the nonexposed people.     

Metabolism and hepatic toxicity of flutamide in cytochrome P450 1A2 knockout SV129 mice

Background Flutamide, a nonsteroidal antiandrogen used for treatment of prostate cancer, causes a temporary increase in transaminase and in some cases severe liver dysfunction. It is dominantly metabolized by cytochrome P450 (CYP) 1A2 into 2-hydroxyflutamide (OH-flutamide), which has stronger antiandrogenic activity without obvious cytotoxicity to cultured hepatocytes. We hypothesized that another subsidiary metabolite might be responsible for induction of hepatotoxicity. Methods Flutamide was administered daily to CYP1A2 knockout mice and parental SV129 mice to compare pharmacokinetics and appearance of hepatic toxicity. Results In the CYP1A2 knockout mice, the plasma concentration of flutamide maintained at a high level and OH-flutamide stayed low; a higher amount of FLU-1, an alternative metabolite of flutamide, was detected in urine. Simple repetitive administration of 800 mg/kg of flutamide for 28 days to CYP1A2 knockout mice did not show abnormal elevation of plasma alanine aminotransferase (ALT). However, after the knockout mice were fed with an amino acid-deficient diet for 2 weeks, which reduced the glutathione (GSH) content to 27% of the initial, administration of 400 mg/kg of flutamide increased ALT to over 200 IU/l and histopathologically moderate hepatitis developed. Since FLU-1 itself did not show cytotoxicity or reduce GSH content in vitro, a further metabolized molecule must cause the hepatotoxicity. Conclusions Blockade of CYP1A2 produced an unknown potential hepatotoxic molecule through FLU-1, and GSH might play an important role in diminishing the reactive hepatotoxic metabolite.     

Factors that influence home return from health care facilities for the elderly--related to the attitude of family caregivers

AIM: The number of users of long-term care insurance has been increasing rapidly since it started in 2000. The number of those who want to enter the long-term care insurance facilities has increased. Although the basic philosophy of long-term care insurance is independence support and self-decision, to enter a facility or home return from facilities is likely to be decided by family caregivers, not by the elderly themselves. Moreover, the number of elderly who return home from welfare facilities is decreasing. We investigated the intension of caregivers who are willing to accept the institutionalized elderly at home and analyzed the factors affecting the acceptance of caregivers. METHODS: Subjects were elderly who were in long-term care insurance facility in June 2004, and their caregivers. The study was conducted between June 2004 and September 2004 in Ibaraki Prefecture in Japan. A face-to-face interview based on a questionnaire was conducted for the institutionalized elderly and by the mail for the caregivers. RESULTS: The caregivers of 34.6% of the elderly who hoped to return home intended to accept them home. There were differences between the plans of the elderly and caregivers. The risk factors (OR, 95% CI) to make the intention of the caregivers to accept the institutionalized elderly home difficult were level of cooperation with other family members to take care of elderly (OR 15.37, 2.05-115.24), dementia behavior disturbance category with more than one (OR 8.34, 1.02-68.05), time spending in bed of a day (OR 1.31, 1.01-1.71), few knowledge of long-term care insurance system of caregivers (OR 3.65,0.81-16.38). CONCLUSION: It has been suggested that more physical activities in the facility, establishment of a care-system for the demented elderly living in the community and an educational campaign by the long-term care insurance system are necessary to increase the willingness of caregivers to accept home return of institutionalized elderly.