Class II LitR serves as an effector of "short" LOV-type blue-light photoreceptor in Pseudomonas mendocina.

PmlR2, a class II LitR/CarH family transcriptional regulator, and PmSB-LOV, a "short" LOV-type blue light photoreceptor, are adjacently encoded in Pseudomonas mendocina NBRC 14162. An effector protein for the "short" LOV-type photoreceptor in Pseudomonas has not yet been identified. Here, we show that PmlR2 is an effector protein of PmSB-LOV. Transcriptional analyses revealed that the expression of genes located near pmlR2 and its homolog gene, pmlR1, was induced in response to illumination. In vitro DNA-protein binding analyses showed that recombinant PmlR2 directly binds to the promoter region of light-inducible genes. Furthermore PmSB-LOV exhibited a typical LOV-type light-induced spectral change. Gel-filtration chromatography demonstrated that the illuminated PmSB-LOV was directly associated with PmlR2, whereas non-illuminated proteins did not interact. The inhibition of PmlR2 function following PmSB-LOV binding was verified by surface plasmon resonance: the DNA-binding ability of PmlR2 was specifically inhibited in the presence of blue light-illuminated-PmSB-LOV. An In vitro transcription assay showed a dose-dependent reduction in PmlR2 repressor activity in the presence of illuminated PmSB-LOV. Overall, evidence suggests that the DNA-binding activity of PmlR2 is inhibited by its direct association with blue light-activated PmSB-LOV, enabling transcription of light-inducible promoters by RNA polymerase.

Abundance of cell-free mitochondrial DNA in spent culture medium associated with morphokinetics and blastocyst collapse of expanded blastocysts.

PURPOSE: This retrospective observational study investigated relationships between the abundance of cell-free mitochondrial DNA (cf-mtDNA) in spent culture medium (SCM) of human-expanded blastocysts and their morphokinetics to address the question of whether the abundance of cf-mtDNA in SCM could predict the quality of blastocysts. METHODS: Embryos (n = 53) were individually cultured in a time-lapse incubator until they reached the expanded blastocyst stage (5 or 6 days), following which copy numbers of cf-mtDNA in SCM (20 µL) of expanded blastocysts were determined using real-time PCR. RESULTS: The duration between start of blastulation to expanded blastocyst (tEB-tSB) and between that of the blastocyst stage to expanded blastocyst (tEB-tB) significantly and positively correlated with the abundance of cf-mtDNA in the SCM (tEB-tSB: r = .46; P < .01; tEB-tB: r = .47; P < .01). The abundance of cf-mtDNA in the SCM was significantly greater in blastocysts with blastocyst collapse (BC), than without BC, and significantly and positively correlated with the number of BC. CONCLUSIONS: The abundance of cf-mtDNA in the SCM was associated with expansion duration and BC. Thus, cf-mtDNA abundance in the SCM serves as a marker to predict the quality of expanded blastocysts.

Analyses of the local control of pulmonary Oligometastases after stereotactic body radiotherapy and the impact of local control on survival.

BACKGROUND: Successful local therapy for oligometastases may lead to longer survival. The purpose of this multicentre retrospective study was to investigate factors affecting the local control (LC) of pulmonary oligometastases treated by stereotactic body radiotherapy (SBRT) and to investigate the impact of LC on survival. METHODS: The inclusion criteria included 1 to 5 metastases, the primary lesion and other extrathoracic metastases were controlled before SBRT, and the biological effective dose (BED10) of the SBRT was 75 Gy or more. The Cox proportional hazards model was used for analyses. RESULTS: Data of 1378 patients with 1547 tumours from 68 institutions were analysed. The median follow-up period was 24.2 months. The one-year, 3-year and 5-year LC rates were 92.1, 81.3 and 78.6%, respectively, and the 1-year, 3-year and 5-year overall survival rates were 90.1, 60.3 and 45.5%, respectively. Multivariate analysis for LC showed that increased maximum tumour diameter (p = 0.011), type A dose calculation algorithm (p = 0.005), shorter overall treatment time of SBRT (p = 0.035) and colorectal primary origin (p < 0.001 excluding oesophagus origin) were significantly associated with a lower LC rate. In the survival analysis, local failure (p < 0.001), worse performance status (1 vs. 0, p = 0.013; 2-3 vs. 0, p < 0.001), oesophageal primary origin (vs. colorectal origin, p = 0.038), squamous cell carcinoma (vs. adenocarcinoma, p = 0.006) and increased maximum tumour diameter (p < 0.001) showed significant relationships with shorter survival. CONCLUSIONS: Several factors of oligometastases and SBRT affected LC. LC of pulmonary oligometastases by SBRT showed a significant survival benefit compared to patients with local failure.

Factors related to primary cancer death and non-primary cancer death in patients treated with stereotactic body radiotherapy for pulmonary oligometastases.

Cancer-specific death (CSD) and non-cancer-specific death (non-CSD) after stereotactic body radiotherapy (SBRT) for pulmonary oligometastases have not been studied in detail. The aim of this study was to determine the cumulative incidences of CSD and non-CSD and to reveal prognostic factors. Data from a large survey of SBRT for pulmonary oligometastases were used for analyses, and patients with unknown cause of death were excluded from current analyses. CSD was primary cancer death and non-CSD was non-primary cancer death including a series of cancer treatment-related deaths. Cumulative incidences were calculated using the Kaplan-Meier method and a stratified Cox regression model was used for multivariate analyses (MVA). Fifty-two patients with an unknown death were excluded and a total of 1326 patients was selected. CSD and non-CSD occurred in 375 and 109 patients, respectively. The median OS period was 53.2 months and the cumulative incidences of 1-, 3-, and 5-year CSD vs. non-CSD rates were 6.5% vs. 2.3%, 29.5% vs. 8.6%, and 41.2% vs. 11.0%, respectively. In MVA, the incidence of CSD was related to performance status (1 vs. 0; p < 0.001, 2-3 vs. 0; p = 0.011), oligometastatic state (sync-oligometastases vs. oligo-recurrence, p = 0.026) and maximum tumor diameter (p = 0.009), and the incidence of non-CSD was related to age (p = 0.001), sex (p = 0.030), performance status (2-3 vs. 0; p = 0.002), and irradiated tumor-located lung lobe (left lower lobe vs. other lobes, p = 0.036). CSD was main cause of death, but non-CSD was not rare after SBRT. Prognostic factors for CSD and non-CSD were different, and an understanding of the factors would help in treatment.

Comparative analysis of cell-free DNA content in culture medium and mitochondrial DNA copy number in porcine parthenogenetically activated embryos.

We examined the effect of ploidy on mitochondrial DNA (mtDNA) copy number in embryos and the amount of cell-free mitochondrial and nucleic DNA content (cf-mtDNA and cf-nDNA) in spent culture medium (SCM). Oocytes collected from the ovaries were matured, activated, incubated in medium containing cycloheximide (CHX) or CHX and cytochalasin B (CB) for 4.5 h to produce haploid or diploid embryos (H-group and D-group embryos). These embryos were cultured for 7 days, and the blastocysts and SCM were examined. The amount of mtDNA and nDNA was determined by real-time PCR. The rate of development to the blastocyst stage was higher for the D-group than for the H-group. Moreover, D-group blastocysts had less mtDNA compared to the H-group blastocysts. After activation, the mitochondrial content was constant before the blastocyst stage in D-group embryos, but increased earlier in H-group embryos. The amount of cf-mtDNA in the SCM of D-group blastocysts was greater than that of H-group blastocysts. However, when the cf-mtDNA in the SCM of 2 cell-stage embryos (day 2 post-activation) was examined, the amount of cf-mtDNA was greater in the H-group than in the D-group embryos. When D-group embryos were cultured for 7 days, a significant correlation was observed between the total cell number of blastocysts and cf-nDNA content in the SCM. Hence, although careful consideration is needed regarding the time point for evaluating mtDNA content in the embryos and SCM, this study demonstrates that mtDNA in the embryos and SCM was affected by the ploidy of the embryos.

Effect of aging on mitochondria and metabolism of bovine granulosa cells.

This study investigated the effect of aging on mitochondria in granulosa cells (GCs) collected from the antral follicles of young and aged cows (25-50 months and over 140 months in age, respectively). When GCs were cultured under 20% O2 for 4 days, mitochondrial DNA copy number (Mt-number), determined by real-time PCR, increased throughout the culture period, and the extent of increase was greater in the GCs of young cows than in those of old cows. In a second experiment, GCs were cultured under 20% O2 for 24 h. Protein levels of TOMM20 and TFAM in GCs were lower in aged cows than in young cows, and the amount of reactive oxygen species and the mitochondrial membrane potential were higher, whereas ATP content and proliferation activity were lower, respectively. Glucose consumption and lactate production were higher in the GCs of aged cows than in those of young cows. When GCs were cultured under 5% or 20% O2 for 24 h, low O2 decreased ATP content and increased glucose consumption in GCs of both age groups compared with high O2; however, low O2 decreased the Mt-number only in the GCs of young cows. In conclusion, we show that aging affects mitochondrial quantity, function, and response to differential O2 tensions in GCs.

Pulmonary Oligometastases Treated by Stereotactic Body Radiation Therapy: A Nationwide Survey of 1,378 Patients.

AIM: This study was performed to confirm the superior overall survival (OS) after pulmonary oligo-recurrence compared to pulmonary sync-oligometastases in a large nationwide study. PATIENTS AND METHODS: Patients that met the following criteria were included: 1 to 5 lung-only metastases at the beginning of stereotactic body radiation therapy (SBRT) was performed between January 2004 and June 2015, and the biological effective dose (BED) of SBRT was 75 Gy or more. The parameters included in the analyses were age, gender, ECOG PS, primary lesion, pathology, oligoetastatic state, SBRT date, chemotherapy before SBRT, chemotherapy concurrent SBRT, chemotherapy after SBRT, maximum tumor diameter, number of metastases, field coplanarity, dose prescription, BED10, OTT of SBRT. RESULTS: In total, 1,378 patients with 1,547 tumors were enrolled. Oligo-recurrence occurred in 1,016 patients, sync-oligometastases in 118, and unclassified oligometastases in 121. The three-year OS was 64.0% for oligo-recurrence and 47.5% for sync-oligometastasis (p<0.001). In the multivariate analysis, the hazard ratio (HR) for sync-oligometastases versus oligo-recurrence was 1.601 (p=0.014). Adverse events of Grade 5 were occurred in 3 patients. CONCLUSION: This is the first nationwide to indicate that the OS of patients with pulmonary oligo-recurrence is better than that of patients with sync-oligometastases.

Improvement of implantation potential in mouse blastocysts derived from IVF by combined treatment with prolactin, epidermal growth factor and 4-hydroxyestradiol.

STUDY QUESTION: Can supplementation of medium with prolactin (PRL), epidermal growth factor (EGF) and 4-hydroxyestradiol (4-OH-E2) prior to embryo transfer improve implantation potential in mouse blastocysts derived from IVF? SUMMARY ANSWER: Combined treatment with PRL, EGF and 4-OH-E2 improves mouse blastocyst implantation rates, while alone, each factor is ineffective. WHAT IS KNOWN ALREADY: Blastocyst dormancy during delayed implantation caused by ovariectomy is maintained by continued progesterone treatment in mice, and estrogen injection rapidly activates blastocysts to implantation-induced status in vivo. While the expression of many proteins is upregulated in implantation-induced blastocysts, selective proteolysis by proteasomes, such as estrogen receptor α (ESR1), occurs in implantation-induced blastocysts to achieve implantation-competent status. It is worth evaluating the proteins expressed during these periods to identify humoral factors that might improve the implantation potential of IVF-derived blastocysts because the poor quality of embryos obtained by IVF is one of the major causes of implantation failure. STUDY DESIGN, SIZE, DURATION: Superovulated oocytes from ICR mice were fertilized with spermatozoa and then cultured in vitro in potassium simplex optimized medium (KSOM) without phenol red (KSOM-P) for 90-96 h. Blastocysts were treated with PRL (10 or 20 mIU/mL), EGF (5 or 10 ng/mL) or 4-OH-E2 (1 or 10 nM) in KSOM-P for 24 h. PARTICIPANTS/MATERIALS, SETTING, METHODS: Levels of breast cancer 1 (BRCA1), EGF receptor (EGFR, also known as ERBB1), ERBB4, tubulointerstitial nephritis antigen-like 1 (TINAGL1) and ESR1 protein were examined with immunohistochemical analysis using immunofluorescence methods and confocal laser scanning microscopy. For embryo transfer, six blastocysts were suspended in HEPES-buffered KSOM-P medium and transferred into the uteri of recipient mice on the morning of Day 4 (0900-1000 h) of pseudopregnancy (Day 1 = vaginal plug). The number of implantation sites was then recorded on Day 6 using the blue dye method. MAIN RESULTS AND THE ROLE OF CHANCE: PRL, EGF and 4-OH-E2 each promoted BRCA1 protein level in the trophectoderm (TE). While PRL treatment resulted in an increase in EGFR, EGF increased both EGFR and ERBB4 in the blastocyst TE. TINAGL1 in the TE was enhanced by 4-OH-E2, which also increased localization of this protein to the basement membrane. Treatment with PRL, EGF or 4-OH-E2 alone did not improve blastocyst implantation rates. Combined treatment with PRL, EGF and 4-OH-E2 resulted in increased levels of EGFR, ERBB4, TINAGL1 and BRCA1 in the TE, whereas ESR1 was not upregulated in the treated blastocysts. Furthermore, combined treatment with PRL, EGF and 4-OH-E2 improved blastocyst implantation rates versus control (P = 0.009). LARGE SCALE DATA: Not applicable. LIMITATIONS, REASONS FOR CAUTION: Our studies were carried out in a mouse model, and the conclusions were drawn from limited results obtained from one species. Whether the increase in EGFR, ERBB4 and TINAGL1 protein in the TE improves implantation potential of blastocysts needs to be further studied experimentally by assessing other expressed proteins. The influence of combined supplementation in vitro of PRL, EGF and 4-OH-E2 on implantation also requires further examination and optimization in human blastocysts before it can be considered for clinical use in ART. WIDER IMPLICATIONS OF THE FINDINGS: Enhanced implantation potential by combined treatment with PRL, EGF and 4-OH-E2 appears to result in the upregulation of at least two distinct mechanisms, namely signaling via EGF receptors and basement membrane formation during the peri-implantation period in mice. While PRL, EGF and 4-OH-E2 each promoted BRCA1 protein level in the TE, treatment with each alone did not improve blastocyst implantation. Therefore, BRCA1 protein appears to be unnecessary for the attachment reaction in blastocysts in mice Combined supplementation of PRL, EGF and 4-OH-E2 might also be of relevance for embryo transfer of human IVF-derived blastocysts for ART. STUDY FUNDING/COMPETING INTEREST(S): This work was supported in part by the JSPS KAKENHI [Grant numbers 22580316 and 25450390 (to H.M.)] and the Joint Research Project of Japan-U.S. Cooperative Science Program (to H.M.). The authors have no conflict of interest to declare.

Degradation of estrogen receptor α in activated blastocysts is associated with implantation in the delayed implantation mouse model.

Implantation of a blastocyst into a receptive uterus involves a series of highly coordinated cellular and molecular events directed by ovarian estrogen and progesterone. In particular, estrogen is essential for on-time uterine receptivity and blastocyst activation in mice. Although estrogen receptor α (ERα) is expressed in blastocysts, its targeted disruption leaves embryonic development and implantation unaffected. Therefore, the role of ERα in implanting blastocysts remains unclear. Using a delayed implantation model in mice, we showed increased expression of ERα in implantation-induced (activated) blastocysts; however, this ERα expression in activated blastocysts decreased within 6-h culture. In contrast, breast cancer 1 (Brca1) was maintained in the blastocysts during the culture. The treatment of activated blastocysts with the proteasome inhibitor MG132 demonstrated that proteolysis is associated with down-regulation of ERα expression in activated blastocysts. Embryo transfer of MG132-treated activated blastocysts into recipient mice on the morning of Day 4 of pseudopregnancy (Day 1 = vaginal plug) showed a decreased implantation rate, whereas combined treatment with MG132 and the ER antagonist, ICI 182,780, resulted in recovery of the rate of implantation. This study has revealed that down-regulation of ERα in activated blastocyst is associated with completion of blastocyst implantation after embryo transfer on the morning of Day 4 of pseudopregnancy. Our results also suggest that selective protein turnover, such as that of ERα, occurs in activated blastocysts, while expression of other proteins, including Brca1, is maintained at the same stage.

[Conduct of survey four months after the start of EMR for home visits to promote cooperation between hospitals and clinics].

Electronic medical records(EMR)for home visits were introduced in October 2013 at our institution in order to ensure smooth cooperation between the hospital and clinic by sharing the details of a patient's medical record. A system was developed for remote desktop connections to the EMR terminal server(virtual server)with the use of an SSL-VPN. Mobile terminals and mobile printers were used. Four months after the start of this system, a survey was conducted for 41 home care professionals and other staff(physicians, nurses, and office staff). Home care staff indicated that they had problems with the system, including bad connections and operating conditions, and difficulties responding to problems when they arose. Other staff indicated that they were able to acquire patient information faster than with paper-based records. Future issues include improvements to the user-friendliness of the terminals and improved responses to problems when they occur.

Pharmacists' awareness and attitude toward blood pressure measurement at home and in the pharmacy in Japan.

The aim of this study is to investigate the status of Japanese pharmacists' awareness and attitude toward blood pressure (BP) measurement at home (HBP) and in the pharmacy. Of the 708 community pharmacists and the 117 hospital pharmacists, more than 90% of pharmacists answered that HBP was equally important to or more important than clinic BP, 71.9% (community) and 48.7% (hospital) recommended HBP measurement to the hypertensive patients, and about 15% correctly recognized the reference values of HBP hypertension. Among community pharmacists, 54.0% answered that BP-measuring devices were available in their pharmacy. More aggressive promotion of HBP measurement among pharmacists is warranted.

Association between fibroblast growth factor 23 and left ventricular hypertrophy in maintenance hemodialysis patients. Comparison with B-type natriuretic peptide and cardiac troponin T.

BACKGROUND: Fibroblast growth factor 23 (FGF-23) is a novel bone-derived phosphate-regulating hormone, and serum FGF-23 levels are associated with mortality among hemodialysis (HD) patients. However, the pathophysiological role of FGF-23 in those patients remains unclear, so the association between serum FGF-23 levels and known cardiac biomarkers or echocardiographic measurements were investigated in long-term HD patients without cardiac symptoms. METHODS AND RESULTS: The 87 consecutive patients treated in a single HD center (51 males, 36 females; mean age 64 years, mean HD duration 5.8 years) were included in this study. Comprehensive echocardiography was performed after HD. Blood samples were obtained before HD. Serum FGF-23 levels in dialysis patients were 1,171±553pg/ml. In univariate analysis, serum phosphate (r=0.443, P<0.001) and calcium levels (r=0.256, P=0.04), left ventricular mass index (LVMI) (r=0.268, P=0.039) were significantly associated with FGF-23 levels. Neither the B-type natriuretic peptide (BNP) nor the cardiac troponin T level was correlated with FGF-23. In multivariate regression analysis, only LVMI (ß=0.287, P=0.031, confidence interval (CI) 0.390-8.040) and phosphate levels (ß=0.419, P=0.001, CI 57.12-207.7) and calcium levels (ß=0.277, P=0.025, CI 24.95-360.1) remained significantly correlated with FGF-23. CONCLUSIONS: Beside BNP, FGF-23 was identified as a factor that is significantly associated with LVMI. FGF-23 could be a novel biomarker of left ventricular overload, which is closely associated with the increased risk of death in HD patients.

Blood pressure-lowering effect and duration of action of bedtime administration of doxazosin determined by home blood pressure measurement.

The effects and duration of action of bedtime administration of doxazosin 2 mg for 4 weeks on uncontrolled morning home hypertension were investigated. Morning home blood pressure (HBP) was significantly lowered by bedtime administration of doxazosin. Doxazosin significantly lowered evening HBP only in the subgroup of patients with an uncontrolled evening HBP. The evening (E)/morning (M) ratio was greater in patients with an uncontrolled evening HBP than in those with a controlled evening HBP. The results suggest that bedtime administration of doxazosin effectively suppresses morning HBP in uncontrolled morning hypertensives and lowers evening HBP in uncontrolled evening hypertensives.

Practice and awareness of physicians regarding casual-clinic blood pressure measurement in Japan.

The optimal procedure for casual-clinic blood pressure (CBP) measurement is outlined in the 2004 Japanese guidelines. We investigated the status of physicians' practices and their awareness of CBP measurement immediately and 4 years after the publication of the guidelines using a questionnaire regarding CBP. This survey was conducted among physicians who attended educational seminars on hypertension in 2004-2005 and in 2007-2008; the questionnaire was distributed, completed and collected just before the start of the seminars. Of the 1966 respondents to the 2004-2005 survey and the 2995 respondents to the 2007-2008 survey, the proportion of physicians who answered that CBP was more important than self-measured BP at home (home BP) was less than 10% in both surveys. The proportion of physicians who used a mercury sphygmomanometer (68.1-75.5%) was higher than those who used an automatic and electronic sphygmomanometer (20.7-29.0%) in both surveys. However, the use of an automatic and electronic sphygmomanometer slightly increased from 20.7% in 2004-2005 to 29.0% in 2007-2008. Physicians who were younger or working in a hospital were less likely to measure CBP using the conditions of the guidelines. Approximately 50% of physicians correctly recognized the reference values of hypertension based on CBP measurement (systolic/diastolic, 140/90 mm Hg) in both surveys. The status of physicians' practice and awareness of CBP measurement varied by physicians' age, specialty and work place and those situations did not change over 4 years, underlying the importance of clarifying factors inherent in these situations.

Practice and awareness of physicians regarding home blood pressure measurement in Japan.

The Japanese Society of Hypertension published guidelines for home blood pressure (HBP) measurement in 2003 and for the management of hypertension in 2004. The objective of this study was to investigate the status of physicians' practice and awareness of HBP measurement based on the Japanese guidelines and compare the status between immediately after and 4 years after publication of the guidelines. A questionnaire survey regarding HBP was conducted among physicians who attended educational seminars on hypertension in 2004-05 and in 2007-08. This questionnaire was distributed, completed, and collected just before the start of the seminars. Of the 1966 and 2995 respondents to the 2004-05 and 2007-08 surveys, respectively, 90.2 and 94.6% recommended HBP measurement to their patients. The majority of physicians recommended use of the upper-arm cuff device, and recommendation of the number of measurements, documentation and evaluation of the measured values varied widely among physicians, both in 2004-05 and in 2007-08. About 10% of physicians showed sufficient understanding of the optimal methods for HBP measurement based on Japanese guidelines both in 2004-05 and in 2007-08. Only 21.6 and 23.9% of physicians correctly recognized the reference values of hypertension based on HBP measurement (systolic/diastolic, 135/85 mm Hg) in 2004-05 and in 2007-08, respectively. Although most Japanese physicians recognized the importance of HBP measurement, many had inadequate knowledge of HBP measurement, both in 2004-05 and in 2007-08. More aggressive promotion of HBP measurement among physicians is warranted.

Influence of home blood pressure measuring conditions in the evening on the morning-evening home blood pressure difference in treated hypertensive patients: the J-HOME study.

OBJECTIVE: The objective of this study was to clarify the factors affecting the morning-evening home systolic blood pressure (BP) difference (home systolic ME dif) in treated hypertensive patients, including evening home BP measuring conditions, based on the data from the Japan Home versus Office BP Measurement Evaluation study. METHODS: The study participants were 3303 essential hypertensive patients (mean age 66.2+/-10.5 years; females 55.3%) treated using antihypertensive drug therapy in primary care settings in Japan. Multivariate regression analysis including the variables that were significantly associated with the home systolic ME dif on the univariate analysis was performed. RESULTS: The mean of the home systolic/diastolic ME dif was 6.1+/-10.8/4.8+/-6.5 mmHg. The proportion of patients measuring evening BP after drinking alcohol was 20.5%, and the proportion of patients measuring evening BP after bathing was 76.8%. Uncontrolled morning systolic BP (morning systolic BP > or =135 mmHg), controlled evening systolic BP (evening systolic BP <135 mmHg), older age (> or =65 years), measurement of evening BP after drinking, and measurement of evening BP after bathing were positively associated with the home systolic ME dif on multivariate regression analysis. CONCLUSION: Measurement of evening BP after drinking and measurement of evening BP after bathing were strongly associated with an increased home systolic ME dif, independent of morning and evening home BP levels. Therefore, evening home BP measuring conditions (before or after drinking alcohol and bathing) should be taken into account while evaluating the home systolic ME dif.

A randomized trial of olfactory stimulation using black pepper oil in older people with swallowing dysfunction.

OBJECTIVES: To determine the effect of olfactory stimulation with volatile black pepper oil (BPO) on risk factors for pneumonia. DESIGN: A 1-month randomized, controlled study. SETTING: Nursing homes in Japan that serve as long-term care facilities for older residents who are physically handicapped, mainly because of cerebrovascular disease. PARTICIPANTS: One hundred five poststroke residents. MEASUREMENTS: Latency of the swallowing reflex (LTSR), the number of swallowing movements, serum substance P (SP), and regional cerebral blood flow (rCBF). RESULTS: Nasal inhalation of BPO for 1 minute shortened LTSR, compared with that of lavender oil and distilled water (P < .03). Compared with the period before the study, the 1-month intervention using BPO improved LTSR with an increase of serum SP (P < .01). The number of swallowing movements for 1 minute during the nasal inhalation of BPO increased (P < .001). Multiple comparisons showed a poststudy increase in rCBF within the insular cortex (P < .001). Compared with the prestudy rCBF, BPO intervention increased rCBF in the right orbitofrontal and left insular cortex (P < .001). CONCLUSION: Inhalation of BPO, which can activate the insular or orbitofrontal cortex, resulting in improvement of the reflexive swallowing movement, might benefit older poststroke patients with dysphagia regardless of their level of consciousness or physical and mental status.

Enhancement of transformed foci and induction of prostaglandins in Balb/c 3T3 cells by palytoxin: in vitro model reproduces carcinogenic responses in animal models regarding the inhibitory effect of indomethacin and reversal of indomethacin's effect by exogenous prostaglandins.

Cell transforming activity of palytoxin, a non-TPA type tumor-promoter, was investigated with the two-stage transformation assay using Balb/c 3T3 cells. Palytoxin showed potent promoting activity; treatment at 1.9 pM or more increased the number of transformed foci after initiation by 3-methylcholanthrene (MCA). Determination of prostaglandin (PG) E2 and PGF(2alpha) concentrations in the culture medium revealed that palytoxin (1.9-3.7 pM for 24 h) stimulated the production of PG in Balb/c 3T3 cells (the concentration reached 3-4 microM), and treatment with PGE2 or PGF(2alpha) itself increased the number of transformed foci of Balb/c 3T3 cells after initiation by MCA. Neither palytoxin nor PGs showed initiating activity. Indomethacin suppressed the promoting activity of palytoxin, but not that of PGE2 and PGF(2alpha). Interestingly, concomitant treatment with PGE2 or PGF(2alpha) in addition to indomethacin markedly reversed the suppressive effect of indomethacin. These findings indicated that the in vitro transformation model could reproduce experiments that have been performed in animal models regarding the inhibitory effect of indomethacin on carcinogenic responses and reversal of indomethacin's effect by exogenous prostaglandin and, therefore, may provide insight into molecular modes of action of palytoxin. In the present study, palytoxin also induced prostaglandin synthesis, and therefore, the Balb/c 3T3 cell model should provide insight into the molecular mechanism by which palytoxin regulates prostaglandin biosynthesis.

[Linear-accelerator-based stereotactic irradiation for metastatic brain tumors].

To assess the safety and availability of stereotactic radiotherapy (SRT) for metastatic brain tumors, we reviewed 54 consecutive cases with a total of 118 brain metastases treated with linear-accelerator-based stereotactic irradiation (STI). Nineteen patients with a total of 27 brain tumors that were larger than 3 cm or close to critical normal tissues were treated with SRT. The marginal dose of SRT was 15-21 Gy (median 21 Gy) in 3 fractions for 3 days. The median marginal dose of stereotactic radiosurgery (SRS) was 20 Gy. Effective rates of imaging studies were 72.7% and 94.4%, and those of clinical symptoms were 46.7% and 55.6% for SRT and SRS, respectively. One-year and two-year survival rates of SRT were 40.9% and 17.6%, respectively, and the median follow-up period was 6.4 months. The one-year survival rate of SRS was 32.7%, with a median follow-up of 4.6 months. Fourteen cases (7 cases each) had recurrent tumors at STI sites. Early complications were observed in one case of SRT and 8 cases of SRS, and late complications occurred in 3 cases of SRS. There were no significant differences among effective rates, survival rates, median follow-up times, recurrence rates, and complications between SRT and SRS. We concluded that SRT is a safe, effective therapy for large or eloquent area metastases.

Multivariate analysis of dissemination relapse of medulloblastoma and estimation of its time parameter for craniospinal irradiation.

PURPOSE: To examine in retrospect prognostic factors influencing meningeal dissemination relapse of medulloblastoma (MB) and to estimate time parameter gamma/alpha of the biologically effective dose for fractionated craniospinal irradiation (CSI). MATERIALS AND METHODS: Fifty-eight patients with MB who had been treated at our six hospitals from 1980 to 1990, were analyzed by the proportional hazards model consisting of radiation factors of both CSI and local irradiation (LI), sequential CSI time-lag, and eight non-radiation factors (gender, age, performance status, T-stage, dissemination score, extent of resection, and use of chemotherapy and immunotherapy). The gamma/alpha for CSI was estimated by the profile likelihood method using the maximum value of conditionally calculated time-incorporated biologically effective dose, tBEDmax, of the field treated with the least dose. RESULTS: Dissemination relapse was seen in 23 (40%) patients. Nineteen disseminations occurred within four years, and the cumulative dissemination-free rate was 64% at five years. The site of initial dissemination relapse was both cranial and spinal in 13 patients (57%). Dissemination relapse was accompanied with local failure in 43% (10/23) of patients, and four of them were seen later than four years. In the multivariate analysis, significant prognostic factors were dissemination score (p=0.0008) and total dose of CSI (p=0.018). The estimate of gamma/alpha for CSI was about 0.2 Gy/day in BED units. In another multivariate analysis including the best-fitted tBEDmax, significant prognostic factors were dissemination score and the tBEDmax of both CSI (p=0.021) and LI (p=0.024). CONCLUSION: This analysis indicated that the dissemination score, total dose of CSI, and tBEDmax of both CSI and LI were significantly prognostic for dissemination relapse of MB. The estimate of gamma/alpha for CSI was smaller than that derived from our previous analysis for LI. However, in order to estimate the time factor for CSI more precisely, a larger group of patients treated with concurrent CSI is needed.