Randomized clinical trial of landiolol hydrochloride for the prevention of atrial fibrillation and postoperative complications after oesophagectomy for cancer.

BACKGROUND: Atrial fibrillation is common after oesophageal surgery. The aim of this study was to evaluate whether landiolol hydrochloride was effective and safe in the prevention of atrial fibrillation after oesophagectomy, and to see whether a reduction in incidence of atrial fibrillation would reduce other postoperative complications. METHODS: This single-centre study enrolled patients scheduled for transthoracic oesophagectomy in a randomized, double-blind, placebo-controlled trial between March 2013 and January 2016. Enrolled patients were randomized with a 1 : 1 parallel allocation ratio to either landiolol prophylaxis or placebo. The primary endpoint was the occurrence of atrial fibrillation after oesophagectomy. Secondary endpoints were incidence of postoperative complications, and effects on haemodynamic and inflammatory indices. RESULTS: One hundred patients were enrolled, 50 in each group. Postoperative atrial fibrillation occurred in 15 patients (30 per cent) receiving placebo versus five (10 per cent) receiving landiolol (P = 0·012). The overall incidence of postoperative complications was significantly lower in the landiolol group (P = 0·046). In the landiolol group, postoperative heart rate was suppressed effectively, but the decrease in BP was not harmful. The interleukin 6 level was significantly lower on days 3 and 5 after surgery in the landiolol group (P = 0·001 and P = 0·002 respectively). CONCLUSION: Landiolol was effective and safe in preventing atrial fibrillation after oesophagectomy. Registration number: UMIN000010648 (http://www.umin.ac.jp/ctr/).

Phase I/II study of divided-dose docetaxel, cisplatin and fluorouracil for patients with recurrent or metastatic squamous cell carcinoma of the esophagus.

Squamous cell carcinoma of the esophagus (SCCE) has a poor prognosis compared with other gastrointestinal cancers. Many patients present with locoregional unresectable or metastatic disease at the time of diagnosis. For these patients with metastatic esophageal cancer, chemotherapy is generally indicated. The aim of this phase I/II study was to evaluate the efficacy and safety of the combined use of docetaxel, cisplatin (CDDP) and 5-fluorouracil (5-FU)(DCF) in patients with recurrent/metastatic SCCE. This study adopted divided doses of docetaxel and CDDP in order to reduce the toxicities of the treatment. The dose of docetaxel was escalated using the following protocol in the phase I stage: level 1, 30 mg/m2; level 2, 35 mg/m2 and level 3, 40 mg/m2, which was intravenously infused for 2 hours on days 1 and 8. CDDP was administered at a dose of 12 mg/m2 infused for 4 hours on days 1-5. The 5-FU was administered at a dose of 600 mg/m2 continuously infused from day 1 to 5. This regimen was repeated every 4 weeks. The study subjects were nine patients (phase I) and 48 patients (phase II). The recommended dose was determined as level 3 in phase I. In the phase II stage, the overall response rate was 62.5%, with a complete response rate of 12.5%. The median progression-free survival was 6 months, and the median overall survival was 13 months. Grade 3/4 toxicities of leukopenia, neutropenia and febrile neutropenia occurred in 64.6%, 68.8% and 14.6% of the patients, while grade 3/4 non-hematological toxicities were relatively rare. No treatment-related death was recorded. This modified DCF regimen with divided doses can be a tolerable and useful regimen of definitive chemotherapy for unresectable SCCE because of its high efficacy, although adequate care for severe neutropenia must be administered.

Randomized clinical trial comparing long-term quality of life for Billroth I versus Roux-en-Y reconstruction after distal gastrectomy for gastric cancer.

BACKGROUND: Patients' quality of life (QoL) deteriorates remarkably after gastrectomy. Billroth I reconstruction following distal gastrectomy has the physiological advantage of allowing food to pass through the duodenum. It was hypothesized that Billroth I reconstruction would be superior to Roux-en-Y reconstruction in terms of long-term QoL after distal gastrectomy. This study compared two reconstructions in a multicentre prospective randomized clinical trial to identify the optimal reconstruction procedure. METHODS: Between January 2009 and September 2010, patients who underwent gastrectomy for gastric cancer were randomized during surgery to Billroth I or Roux-en-Y reconstruction. The primary endpoint was assessment of QoL using the Functional Assessment of Cancer Therapy - Gastric (FACT-Ga) questionnaire 36 months after surgery. RESULTS: A total of 122 patients were enrolled in the study, 60 to Billroth I and 62 to Roux-en-Y reconstruction. There were no differences between the two groups in terms of postoperative complications or mortality, and no significant differences in FACT-Ga total score (P = 0·496). Symptom scales such as epigastric fullness (heaviness), diarrhoea and fatigue were significantly better in the Billroth I group at 36 months after gastrectomy (heaviness, P = 0·040; diarrhoea, P = 0·046; fatigue, P = 0·029). The rate of weight loss in the third year was lower for patients in the Billroth I group (P = 0·046). CONCLUSION: The choice of anastomotic reconstruction after distal gastrectomy resulted in no difference in long-term QoL in patients with gastric cancer. REGISTRATION NUMBER: NCT01065688 (http://www.clinicaltrials.gov).

A rapid monitoring system of human herpesviruses reactivation by LightCycler in stem cell transplantation.

To establish a practical monitoring system of human herpesviruses reactivation in patients undergoing stem cell transplantation, we developed a new, very rapid, highly sensitive, and quantitative PCR assay for accurate measurement of human cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and Epstein-Barr virus (EBV) DNA using LightCycler. The LightCycler system revealed that there was a linear correlation in the wide range of viral template DNA at the indicated number of PCR cycles. Peripheral blood cells were collected from 16 patients undergoing stem cell transplantation. The cut-off level of CMV and HHV-6 was assessed as 10(2) copies/microg and that of EBV as 10(3). High numbers of CMV genomes were detected in 3/13 patients after transplant, and reactivation of HHV-6 was frequently seen, whereas none of the patient showed an elevation of EBV genome copies until the end of the observation period. In the present study, the reactivation of beta herpesviruses is associated with the occurrence of thrombotic microangiopathy (TMA) in two patients undergoing allogeneic BMT. Therefore, it may contribute in clarifying the pathological potential of human herpesviruses using a large number of clinical samples. Our results suggest that this system may be useful for monitoring viral reactivation.

Validity and reliability of single-item questions about physical activity.

The Japan Collaborative Cohort Study for Evaluation of Cancer Risk Sponsored by Monbusho (JACC Study) included in its self-administered questionnaires some single-item questions concerning physical activity. We examined the validity of the questions among 1,730 Japanese adults and the reliability of the questions among 1,075 Japanese adults. The validity of the sports and physical exercise questions was estimated by comparing the self-administered questionnaire responses with the time spent on the activity and the energy expenditure index for the previous 12-month period, elicited by the interviewing method used in the Japan Lifestyle Monitoring Study with a minor modification. The Spearman's rank correlation coefficients ranged from 0.43 to 0.60, showing moderate correlations. On the other hand, test-retest reliability was estimated by comparing the responses from two separate surveys conducted roughly one year apart. Weighted kappa coefficients of sports and physical exercise questions, classified according to sex and age, ranged from 0.39 to 0.56, showing moderate reliability; and those of a question about walking ranged from 0.25 to 0.39, showing fair reliability. We suggest that measuring physical activity level with these single-item questions may be appropriate for establishing baseline data that reflects long-term physical activity in a large-scale cohort study targeting lifestyle-related diseases.

(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assays showed a lack of toxicity of ganciclovir (GCV), C.OXTs, and their derivatives, to TaY(OK) cells at 1 microM. Therefore we compared the antiviral potencies of these drugs at 1 microM by monitoring the viral loads produced during a 1-day period during the course of the drug treatment. Among the drugs tested, 3'-fluorocarbocyclic oxetanocin A (3'F-C.OXT-A) was the most effective for inhibiting the virus production, and at concentrations ranging from 0.5 microM to 10 microM, the inhibition of the viral production was dose-dependent. A comparison of the chemical structures of the derivatives with that of C.OXT-A, which is the parental molecule, suggested that the 3'-fluorine-modification might account for the higher anti-HHV-6 activity and lower cytotoxicity.

An infection of TaY cells, which originated from an adult T-cell leukemia, with an HHV-6B OK isolate resulted in a chronically infected culture, termed TaY(OK). Cell cloning analysis revealed that the TaY(OK) culture consisted of a mixture of cells permissive and refractory to the infection, and that the permissive cells were continuously produced from the refractory cell population. Since the chronically infected culture has been maintained for over 2 years without the addition of uninfected TaY cells, we used it for an evaluation of the antiviral potency of nucleoside analogs, especially carbocyclic oxetanocins (C.OXTs).

Amino acid sequence of squid troponin C.

The complete amino acid sequence of squid Todarodes pacificus troponin C (TnC), which was shown to bind only 1 mol Ca(2+)/mol, was determined by both the Edman and cDNA methods. The squid TnC is composed of 147 amino acids including an unblocked Pro at the N-terminus and the calculated molecular weight is 17003.9. Among the four potential Ca(2+)-binding sites, namely sites I-IV from the N-terminus, only site IV completely satisfied the consensus amino acid sequence for the active Ca(2+)-binding loop. This indicates that squid TnC possesses a single Ca(2+)-binding site at the site IV as scallop TnCs [Nishita et al., J. Biol. Chem. 269 (1994) 3464-3468; Ojima et al., Arch. Biochem. Biophys. 311 (1994) 272-276). The sequence homology of squid TnC to TnCs of scallop, arthropods, and rabbit was 61%, 31-38%, and 31%, respectively. In the sequence of the central D/E-helix region of squid and scallop TnCs, a deletion of three amino acids was required to maximize the homology with the other TnCs.

Functional role of Ca(2+)-binding site IV of scallop troponin C.

Scallop troponin C (TnC) binds only one Ca(2+)/mol and the single Ca(2+)-binding site has been suggested to be site IV on the basis of the primary structure [K. Nishita, H. Tanaka, and T. Ojima (1994) J. Biol. Chem. 269, 3464-3468; T. Ojima, H. Tanaka, and K. Nishita (1994) Arch. Biochem. Biophys. 311, 272-276]. In the present study, the functional role of Ca(2+)-binding site IV of akazara scallop (Chlamys nipponensis akazara) TnC in Ca(2+)-regulation was investigated using a site-directed mutant with an inactivated site IV (TnC-ZEQ), N- and C-terminal half molecule mutants (TnC(N) and TnC(C)), and wild-type TnC (TnC(W)). Equilibrium dialysis using (45)Ca(2+) demonstrated that TnC(W) and TnC(C) bind 0.6-0.8 mol of Ca(2+)/mol, but that TnC-ZEQ and TnC(N) bind virtually no Ca(2+). The UV difference spectra of TnC(W) and TnC(C) showed bands at around 280-290 nm due to the perturbation of Tyr and Trp upon Ca(2+)-binding, while TnC-ZEQ and TnC(N) did not show these bands. In addition, TnC(W) and TnC(C) showed retardation of elution from Sephacryl S-200 upon the addition of 1 mM CaCl(2), unlike TnC-ZEQ and TnC(N). These results indicate that Ca(2+) binds only to site IV and that Ca(2+)-binding causes structural changes in both the whole TnC molecule and the C-terminal half molecule. In addition, TnC(W), TnC-ZEQ, and TnC(C), but not TnC(N), were shown to form soluble complexes with scallop TnI at physiological ionic strength. On the other hand, the Mg-ATPase activity of reconstituted rabbit actomyosin in the presence of scallop tropomyosin was inhibited by scallop TnI and recovered by the addition of an equimolar amount of TnC(W), TnC-ZEQ, or TnC(C), but not TnC(N). These results imply that the site responsible for the association with TnI is located in the C-terminal half domain of TnC. Ternary complex constructed from scallop TnT, TnI, and TnC(W) conferred Ca(2+)-sensitivity to the Mg-ATPase of rabbit actomyosin to the same extent as native troponin, but the TnC(N)-TnT-TnI and TnC-ZEQ-TnT-TnI complexes conferred no Ca(2+)-sensitivity, while the TnC(C)-TnT-TnI complex conferred weak Ca(2+)-sensitivity. Thus, the major functions of scallop TnC, such as Ca(2+)-binding and interaction with TnI, are located in the C-terminal domain, however, the full Ca(2+)-regulatory function requires the presence of the N-terminal domain.

Use of real-time PCR to monitor human herpesvirus 6 reactivation after allogeneic bone marrow transplantation.

Reactivation of human herpesvirus 6 (HHV-6) is common following allogeneic marrow transplantation, however, little is known about the immune control and pathogenic potential of HHV-6 infection after transplantation. In order to determine whether reactivation of HHV-6 plays an important role in the development of complications in patients undergoing allogeneic bone marrow transplantation or not, we developed a very rapid quantification of viral DNA using a LightCycler. The amount of viral DNA was determined using a supernatant of a chronically infected cell line [TaY(OK)] which contains a known amount of viral DNA. Peripheral blood cells were collected from 5 patients undergoing allogeneic bone marrow transplantation once before transplant and once per week after transplant for 8-24 weeks. The real-time PCR system revealed that there was a linear correlation in the range of 101 to 105 molecules of reference. Using this system, we have found the presence of non-diagnosed HHV-6 reactivation as well as symptomatic infection, indicating the potential for routine implementation of this technology for laboratory diagnosis of HHV-6 infection. Our study shows that this method of rapid quantification of HHV-6 genomes by the real-time PCR using a LightCycler may be useful not only to understand the reconstitution of the immune system following marrow transplantation but also to manage the care of patients.

Effects of removal and reconstitution of myosin regulatory light chain and troponin C on the Ca(2+)-sensitive ATPase activity of myofibrils from scallop striated muscle.

In order to examine the involvement of troponin-linked Ca(2+)-regulation, in addition to well-known myosin-linked Ca(2+)-regulation, in the contraction of molluscan striated muscle, myofibrils from Ezo-giant scallop striated muscle were desensitized to Ca(2+) by removing both myosin regulatory light chain and troponin C by treatment with a strong divalent cation chelator, CDTA. The ATPase level in the desensitized myofibrils was about half the maximum level in intact myofibrils regardless of the Ca(2+)-concentration at 25 and 15 degrees C. In the absence of Ca(2+), the ATPase of the desensitized myofibrils was suppressed by myosin regulatory light chain but not affected by troponin C at either temperature. The ATPase was activated at higher Ca(2+)-concentrations by both myosin regulatory light chain and troponin C, but the activating effects of these two proteins were affected differently by temperature. The activation of ATPase by myosin regulatory light chain was much greater than that by troponin C at 25 degrees C, whereas the activation by troponin C was much greater than that by myosin regulatory light chain at 15 degrees C. The maximum activation was only obtained in the presence of both myosin regulatory light chain and troponin C at these temperatures. These findings strongly suggest that the contraction of scallop striated muscle is regulated through both myosin-linked and troponin-linked Ca(2+)-regulation, and that the troponin-linked Ca(2+)-regulation is more significant at lower temperature.

Association between health-related knowledge and the awareness of blood pressure readings. Japan Lifestyle Monitoring Study Group.

This study aims to evaluate the relationships between awareness of blood pressure testing and health-related knowledge. Among 9 blocks in Japan, one or two areas were selected and a questionnaire survey including physical activity, nutrient intake, drinking habits, smoking habits, medical history, annual frequency of blood pressure testing, subjective sense of wellness, and health related knowledge was conducted. Respondents' sex and age were adjusted as potential confounding factors and the association between possession of health-related knowledge and awareness of blood pressure testing were examined by using a multiple logistic regression model. Among the 1,834 effective responses, 263 (14.3%) answered that they could state their blood pressure levels. These individuals tended to have a greater health-related knowledge. Even after adjusting for other potential confounders (smoking habits, medical history, annual frequency of blood pressure testing, and subjective sense of wellness) health-related knowledge were significantly related to blood pressure awareness.

Efficient propagation of human herpesvirus 6B in a T-cell line derived from a patient with adult T-cell leukemia/lymphoma.

The efficient propagation of the OK strain of the B variant of human herpesvirus 6 (HHV-6B) was demonstrated in a line of T cells, TaY, established from the peripheral blood lymphocytes of a patient with adult T-cell leukemia/lymphoma (ATL). Growth of TaY cells depends on the presence of IL-2 and the cells harbor HTLV-I genomes. A severe cytopathic effect (CPE) was observed in many HHV-6B(OK)-infected TaY cells one week after infection. The release of virus from HHV-6B(OK)-infected TaY cells [TaY(OK)] was first detected after three days and increased rapidly for up to seven days after infection, as demonstrated by PCR. The titer of HHV-6B(OK) in the supernatant was comparable to the value of 10(3.5) TCID50/ml obtained with PHA-activated cord blood lymphocytes (CBL) that had been infected with HHV-6B(OK). The replication of the virus was shown to depend to a considerable extent on cell viability. Electron microscopy revealed many herpesvirus-type capsid- and enveloped-viruses in the nuclei and cytoplasm of degenerated cells in TaY(OK) cultures. The U1102 strain of HHV-6A and the Z29 strain of HHV-6B also infected TaY cells productively, as detected by PCR and an immunofluorescence test. These results suggest that the activation of CD4+ T lymphocytes with mitogens such as PHA or IL-2 and the expression of some cellular gene or the HTLV-I gene might be essential for efficient propagation of HHV-6B. TaY cells should play an important role in future investigations of cell-virus interactions and genetic variations or cell tropism of HHV-6 isolates since no cell line that shows propagation of both HHV-6A and HHV-6B has been reported to date.

Quantification of human herpesvirus 6 in healthy volunteers and patients with lymphoproliferative disorders by PCR-ELISA.

To determine whether actual numbers of human herpesvirus 6 (HHV-6) genome in hematologic neoplasias are associated with disease condition, we developed a quantitative PCR-ELISA for detection of HHV-6. The amount of viral DNA was determined using externally amplified known amounts of the plasmid DNA containing the viral target sequences. First, we determined a viral burden in peripheral blood leukocytes obtained from 23 healthy volunteers and four specimens of lymph nodes with reactive hyperplasia. Using 1 microg of DNA, the prevalence of HHV-6 was 43.4% (10/23), ranging from 0 to 100 HHV-6 genomes in blood obtained from healthy volunteers. The amounts of HHV-6 genomes were < 10 in four non-neoplastic lymph node specimens. We next examined the amount of viral DNA in 21 blood specimens and 19 lymph node specimens obtained from patients with lymphoproliferative diseases (LPD) at the time of diagnosis. The number of HHV-6 genomes in most of the B-cell lymphoma was < 5 in both blood and lymph node specimens, however, two lymph node specimens obtained from immunoblastic lymphadenopathy (IBL) and T-cell lymphoma had very high levels of HHV-6 viral DNA (3705 and 810, respectively). We also found that HHV-6 genomes in peripheral blood were more than 1000 in two patients with chronic lymphocytic leukemia. For all LPD patients combined, there were significantly higher levels of viral DNA (200.6 +/- 654.8 HHV-6 genomes per 1 microg purified DNA) compared to those in healthy volunteers (10.0 +/- 21.0 HHV-6 genomes per 1 microg purified DNA) (P < 0.05). This study demonstrates that a high level of HHV-6 viral DNA is occasionally associated with LPD patients. Although it is still uncertain whether HHV-6 is related to the pathogenesis in LPD or not, our results suggest that measurement of HHV-6 genomes using PCR-ELISA may be useful not only to understand the mechanism of HHV-6 infection in hemopoietic neoplasia but also to manage the care of immnocompromised patients such as bone marrow transplant patients.

[Ethical issues regarding individual data collection and utilization in community health care programs].

To understand the nature of ethical issues in community-based health care programs, we conducted a mail survey of subjects who were public health nurses employed by municipal governments. The questionnaire consisted of questions about data collection, usage, disclosure, and educational experience. In 1996 we received 536 completed questionnaires which were then analyzed. Regarding who should input data into computers, the number of those who considered that municipal offices other than public health nurses would be the most appropriate for the input of examination data was the largest, followed by those who felt that contracting out was best. Many of the public health nurses considered that they needed to obtain informed consent for collection, usage and disclosure of sensitive items, such as data on HIV infection. The number of those considering that they could not disclose results of examinations to other community-based specialists in health and welfare without the subject's agreement was very high. In health examination programs, the public health nurses requested information on date of birth and occupations, but there was some hesitation in requesting the latter information. Although about a half of subjects responded that they did not require data concerning the first sexual intercourse in cervical cancer screening, 90 percent asked breast feeding history in breast cancer screening. Approximately 90 percent gave results of the examination to participants personally through personal communication or mail. Of the respondents, 40 percent reported having had educational courses on ethics while the others did not. There were some responses that reminded us of the unsatisfactory level of understanding about ethical issues, which underlined the need to emphasize importance, of including this in educational curricula.

Multilobular cavernous malformation: report of two cases.

The authors report two cases of cavernous malformation characterized by a multilobular appearance on magnetic resonance images. At surgery, the malformations consisted of several nests of angiomatous components that were separated by intervening brain tissue and connected with each other by tiny vessels. This basic configuration seems to explain the unexpected postoperative recurrence of cavernous malformations and/or rebleeding from the residual lesions.

Analyses of factors influencing participation in the cervical cancer screening programme in the community in Japan.

The association between the participation rates of cervical cancer screening programme and implementation methods, concerning data management, participant convenience, promoting participation, and payment, were analyzed. The data regarding the implementation methods were obtained in a nation-wide questionnaire survey. The relationship between the participation rates and implementation methods were assessed using the chi-square test and multiple logistic regression analyses. In small municipalities, with a population < 10,000 items concerning data management, enlisting the assistance of community organizations, fee exemption, and early morning screening were positively associated with the participation. In middle-sized municipalities, with the population 10,000-49,999, early morning screening, community organizations, items concerning data management, and sending out letters were positively associated with participation. Saturday/Sunday screening, community organizations, letters, and postcards were positively associated with the participation of the older group (> or = 50 years) in large municipalities with population of > 50,000. These results indicate that enlisting the assistance of community organizations and establishing a well-organized data management system are likely to improve participation regardless of municipality size. Other implementation methods must be selected taking into account factors such as municipality population size, as well as the age distribution and characteristics of the target population.

Troponin from smooth adductor muscle of Ezo-giant scallop.

Troponin which can confer Ca2+-sensitivity upon rabbit actomyosin Mg-ATPase activity has been prepared from the smooth adductor muscle of Ezo-giant scallop (Patinopecten yessoensis). The troponin comprises 40-, 20-, and 19-kDa components. In order to characterize the components, they were separated from each other by CM-Toyopearl column chromatography in the presence of 6 M urea. Consequently, the 20-kDa component was identified as troponin C, based on the Ca2+-binding ability. The amount of Ca2+ bound to the troponin C was estimated to be 0.75 mol/mol at 10(-4) M Ca2+ by the equilibrium dialysis method. The 19-kDa component was identified as troponin I on the basis of not only its inhibitory effect on rabbit actomyosin Mg-ATPase activity along with the smooth adductor tropomyosin, but also the releasing effect of the smooth adductor troponin C on the inhibition. On the other hand, the 40-kDa component was regarded as troponin T on the basis that it bound to F-actin-tropomyosin filament and was indispensable for conferring Ca2+-sensitivity upon rabbit actomyosin Mg-ATPase activity, along with troponin C and troponin I. The above assignments were confirmed by both amino acid analysis and immunoblotting using rabbit antisera raised against counterparts of scallop striated muscle troponin.

Family history evaluation in epidemiologic studies; an evaluation of bias in the conventional methods and a proposal of a new one.

We discuss problems of conventional methods to evaluate family histories as risk factors in epidemiologic studies, and propose a new one. In the proposal, incidence rates of the disease interested among family members are treated as exposure levels. Besides, we show the quantitative difference between the results obtained by the conventional methods and the new one using models. Because of not so much information required for the new method, we recommend it even though the difference was not so large.