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This study designs a low-power photoplethysmography (PPG) sensor based on the error compensation method for heartbeat interval acquisition. To perform heartbeat monitoring in daily life, it is necessary to obtain long-term and accurate heartbeat interval data with low power consumption, because of the limited size and battery capacity of the PPG sensor. Effective reduction in the power consumption of the sensor requires the duty-cycled LEDs and lowering pulse repetition frequency (PRF), i.e., decreasing the sampling rate. However, these methods reduce the accuracy of the heartbeat interval measurement because of signal-to-noise ratio (SNR) degradation and sampling errors. We propose an algorithm for heartbeat interval error compensation and incorporate a low-noise readout circuit to improve SNR. The readout circuit uses current integration to achieve low duty-cycle LED driving. A correlated double sampling (CDS) is introduced to minimize the random noise arising from the switching operation of the integration circuit. An error compensation method based on the PPG waveform similarity is also introduced using the autocorrelation and linear interpolation. The measurement results obtained from nine subjects show that a total current consumption of 28.2 μA is achieved with a 20-Hz PRF and 0.3% LED duty cycle. The proposed design effectively reduces the mean absolute error (MAE) of the heartbeat interval to an average of 6.2 ms.

During irradiation sessions for brain tumors or head and neck cancers, some patients experience abnormal olfactory sensations. To date, the frequency of such sensations during these treatment sessions has not been investigated. We analyzed abnormal olfactory sensations in patients who underwent radiation therapy at our institution for primary brain tumors, excluding malignant lymphoma, between January 2009 and January 2018. A total of 191 patients who were awake during radiation treatment and capable of communicating were analyzed in this retrospective medical study. Of these patients, 7 were aware of olfactory sensations during irradiation. The median age of these 7 patients was 13 (range 8-47) years, Six were <20 years of age, accounting for 10% of the total population of similar age (n = 60). However, only 1 of 131 patients aged ≥20 years complained of strange olfactory sensations. Four of seven patients had germ cell tumors, but none had a medulloblastoma. We investigated patients who experienced light sensation, as an internal standard to ascertain the accuracy of this study. Only 10 patients experienced light sensation during their irradiation sessions. This suggests that the frequency of these sensations was possibly underestimated in our study. In conclusion, a considerable number of patients experienced unusual olfactory sensations during radiation treatment. Further prospective studies on abnormal olfactory sensations during irradiation are needed to clarify the underlying mechanism of this sensation., (© The Author(s) 2019. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Objectives: Most patient-derived pancreatic ductal adenocarcinoma (PDAC) xenografts have been established from surgical specimens of patients who have not received chemotherapy. However, xenografts have rarely been established from chemotherapy-resistant, advanced PDACs, because such cases are usually inoperable. The purpose of this study is to establish patient-derived xenografts using PDAC cells refractory to chemotherapy., Methods: Clinical PDAC cells obtained from ascites of patients who had received continuous chemotherapy were implanted into the flanks of immunocompromised mice. Growth and histological features of the xenografts with and without gemcitabine treatment were then analyzed., Results: Ascites-derived PDAC cells were successfully expanded through serial xenograft passage without changes in histological appearance. While treatment with gemcitabine substantially inhibited the growth of all PDAC xenografts tested, the tumor volume gradually increased, and the tumors showed marked regrowth even under continued gemcitabine treatment. These findings are consistent with the actual clinical course of the corresponding patients for each xenograft., Conclusions: Ascites-derived xenograft models represent a valuable experimental system for testing the efficacy of currently available therapeutic compounds on chemotherapy-resistant PDAC cells and for elucidation of the mechanisms underlying chemotherapy resistance.

Attenuated strains of avian Eimeria parasites, generated by the selection of precocious lines through serial passaging in chicks, have been used widely as live vaccines. Detailed morphological transitions including their life cycle depending on the passages remain poorly understood. Here, we showed early development and acceleration of transitions in morphological forms of the asexual schizonts of E. tenella that had been attenuated for virulence by serial passaging. Our results may be helpful in understanding parasitism, facilitating further molecular analyses such as comparative genomic or transcriptomic tests., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Simultaneous evaluation of coagulation and fibrinolysis facilitates an overall understanding of normal and pathological haemostasis. We established an assay for assessing clot formation and fibrinolysis simultaneously using clot waveform analysis by the trigger of a mixture of activated partial thromboplastin time reagent and an optimized concentration of tissue-type plasminogen activator (0·63 μg/ml) to examine the temporal reactions in a short monitoring time (<500 s). The interplay between clot formation and fibrinolysis was confirmed by analysing the effects of argatroban, tranexamic acid and thrombomodulin. Fibrinogen levels positively correlated with coagulation and fibrinolytic potential and initial fibrin clot formation was independent of plasminogen concentration. Plasminogen activator inhibitor-1-deficient (-def) and α2-antiplasmin-def plasmas demonstrated different characteristic hyper-fibrinolytic patterns. For the specificity of individual clotting factor-def plasmas, factor (F)VIII-def and FIX-def plasmas in particular demonstrated shortened fibrinolysis lag-times (FLT) and enhanced endogenous fibrinolysis potential in addition to decreased maximum coagulation velocity, possibly reflecting the fragile formation of fibrin clots. Tranexamic acid depressed fibrinolysis to a similar extent in FVIII-def and FIX-def plasmas. We concluded that the clot-fibrinolysis waveform analysis technique could sensitively monitor both sides of fibrin clot formation and fibrinolysis, and could provide an easy-to-use assay to help clarify the underlying pathogenesis of bleeding disorders in routine clinical practice., (© 2019 British Society for Haematology and John Wiley & Sons Ltd.)

Reduced-intensity stem cell transplantation (RIST) minimizes the adverse effects of traditional hematopoietic stem cell transplantation, and low-dose total-body irradiation (TBI) is administered over a short period prior to RIST (TBI-RIST). Different institutes adopt different approaches for the administration of TBI-RIST, and since no study had previously investigated this issue, a survey of the TBI schedules in Japan was conducted. In October 2015, the Japanese Radiation Oncology Study Group initiated a national survey of TBI-RIST procedures conducted between 2010 and 2014. Of 186 institutions performing TBI, 90 (48%) responded to the survey, 78 of which performed TBI-RIST. Of 2488 patients who underwent TBI for malignant disease at these institutions, 1412 (56.8%) patients were treated for leukemia, 477 (19.2%) for malignant lymphoma, 453 (18.2) for myelodysplastic syndrome, 44 (1.8%) for multiple myeloma, and 102 (4.1%) for other malignant diseases. Further, 206 (52.0%) of 396 patients (a high proportion of patients) who underwent TBI for benign disease had aplastic anemia. The TBI-RIST equipment and treatment methods were similar to those used for myeloablative regimens. Routinely shielded organs included the lungs (43.6%), eyes (50.0%) and kidneys (10.2%). The ovaries (14.1%), thyroid (6.4%) and testicles (16.7%) were also frequently shielded, possibly reflecting an emphasis on shielding reproductive organs in children. TBI-RIST was performed more frequently than myeloablative conditioning in patients with benign disease. Genital and thyroid shielding were applied more frequently in patients treated with TBI-RIST than in patients treated with myeloablative conditioning. In conclusion, this study indicates the status of TBI-RIST in Japan and can assist future efforts to standardize TBI-RIST treatment methods and to design a future multicenter collaborative research study., (© The Author(s) 2019. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.)

Gastrointestinal parasites including Eimeria spp. are known to affect domestic animal productivity causing watery or lethal bloody diarrhea. However, there are few reports on the detailed distribution of bovine Eimeria spp. in cattle, particularly in developing tropical and sub-tropical areas. Using a total of 289 fecal samples collected from beef cattle on Java Island, one of the five main islands of Indonesia, fecal examinations by the Whitlock and sugar flotation methods and molecular surveys were conducted to reveal the prevalence of 6 Eimeria spp. As a result of morphological screening using Whitlock methods and sugar flotation, Eimeria spp. prevalences of 9.4% and 52.3% were confirmed, respectively. The prevalence was higher in younger cattle [under 1 year (63.9%), 1-2 years (75.0%) and more than in 2 year old cattle (42.3%)]. The prevalences of identified species were as follows: 10.4% for E. bovis, 2.8% for E. ellipsoidalis, 2.1% for E. alabamensis, 1.4% for E. zuernii, 1.1% for E. auburnensis, and 0.4% for E. cylindrica. Moreover, prevalences of 12.8% for Strongyloides papillosus, 7.3% for Trichuris globulosa, and 0.3% for Capillaria bovis were detected. Although the average number of oocysts per gram of feces was <100 among the positive samples, and cases of heavy infection were limited, the endemicity of these pathogenic Eimeria species among farms in Indonesia should be noted., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Strangles is a commonly diagnosed and important infectious disease of equids worldwide, caused by Streptococcus equi subsp. equi. We determined the SeM genotypes of S. equi isolated from imported horses at the Japanese border within the past 8 years, which allowed us to classify 12 strains isolated from these horses from each exporter into four allelic groups. These alleles were different from the alleles of past isolates found in Japan. Furthermore, four strains classified into the same allele were isolated from horses from one exporter over several years. In this study, S. equi isolates from different exporters had different SeM alleles. Attention to the hygiene status of farms will be necessary to prevent the incursion of strangles.

Since metastasis accounts for the majority of cancer-associated deaths, studies on the mechanisms of metastasis are needed to establish innovative strategies for cancer treatment. We previously reported that melanoma cell adhesion molecule (MCAM) functions as a critical receptor for S100A8/A9, and binding of S100A8/A9 to MCAM results in the migration of melanoma cells to lung tissue. However, the critical role of MCAM in the original melanoma skin lesion is still not clear. In this study, we aimed to determine the importance of the S100A8/A9-MCAM axis in melanoma dissemination in a skin lesion as a critical early step for metastasis. Mechanistic studies revealed the downstream signaling of MCAM that signaled the induction of metastasis. S100A8/A9-MCAM binding activates mitogen-activated protein kinase kinase kinase 8 (MAP3K8), also termed TPL2, leading to strong activation of the transcription factor ETV4 and subsequent induction of matrix metalloproteinase-25 (MMP25), and finally to induction of melanoma lung tropic metastasis. Collectively, our results demonstrate a crucial role of the S100A8/A9-MCAM signaling axis in metastatic onset of melanoma cells and indicate that strategies targeting the identified pathway may be useful for the establishment of innovative anti-cancer therapies., (Copyright © 2019 The Author(s). Published by Elsevier B.V. All rights reserved.)

We developed a new head supporting device to provide accurate correction of rotational setup during image-guided radiation therapy (IGRT), evaluating its correction performance and the efficacy of dose distribution in stereotactic radiotherapy (SRT) using a helical tomotherapy (HT) system. The accuracy of rotational motion was measured using an electronic inclinometer; we compared device angles and measurement values from 0.0° to 3.0°. The correction accuracy was investigated based on the distance between rotational centers in the device and on megavoltage computed tomography (MVCT); the correction values were compared using distances in the range of 0.0-9.0 cm using a head phantom with a rotational error of 1.5°. For an SRT with a simultaneous integrated boost plan and a rotational error of 3.0° in yaw angle using a head phantom, and for a single-isocenter SRT for multiple brain metastases in the data of three patients, dosimetric efficacy of the HT unit was evaluated for calculated dose distributions with MVCT after rotational correction. This device can correct pitch and yaw angles within 0.3° and can be corrected to within 0.5° for each rotational angle according to the result of MVCT correction regardless of the rotational center position. In the head phantom study, the device had a beneficial impact on rotational correction; D99% for the target improved by approximately 10% with rotational correction. Using patient data with the device, the mean difference based on the treatment planning data was 0.3% for D99% and -0.1% for coverage index to the target. Our rotational setup correction device has high efficacy, and can be used for IGRT., (© 2019 The Authors. Journal of Applied Clinical Medical Physics published by Wiley Periodicals, Inc. on behalf of American Association of Physicists in Medicine.)

The purpose of this study was to quantify actual patient organ doses from megavoltage computed tomography (MVCT) using an MVCT beam model of a helical tomotherapy unit in a general treatment planning system (TPS). Dosimetric parameters (percentage depth dose, lateral beam profile, and longitudinal beam profile) of the MVCT beam were measured using Gafchromic EBT3 films (ISP Corporation, Wayne, NJ, USA) and used for beam modeling in a Pinnacle3 TPS (Philips, Amsterdam, Netherlands); this TPS is widely used with linear accelerators. The created beam model was adjusted and validated by assessing point doses in a cylindrical phantom in static and helical beam plans with fine, normal and coarse pitches. Maximum doses delivered to important organs from MVCT delivery for five clinical cases were calculated using the created beam model. The difference (average ± one standard deviation for all evaluation points) between calculated and measured doses was -0.69 ± 1.20% in the static beam plan. In the helical beam plan, the differences were 1.83 ± 2.65%, 1.35 ± 5.94% and -0.66 ± 8.48% for fine, normal and coarse pitches, respectively. The average maximum additional dose to important organs from MVCT in clinical cases was 0.82% of the prescribed dose. In conclusion, we investigated a method for quantifying patient organ dose from MVCT delivery on helical tomotherapy using an MVCT beam model in a general TPS. This technique enables estimation of the patient-specific organ dose from MVCT delivery, without the need for additional equipment., (© The Author(s) 2019. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.)

To evaluate the accuracy of hemagglutination inhibition (HI) test as the index of feline panleukopenia virus (FPV)-protective ability, sera from 153 FPV-vaccinated cats aged ≥7 months with HI titer of <1:10-1:40, were examined for serum neutralizing (SN) antibody. SN antibody was detected (≥1:10) in 33 (62.3%) of 53 HI antibody-negative cats, and ranged <1:10-1:160. This suggests that FPV-antibody detection sensitivity of HI test is lower than SN test, and SN test is more suitable for the assessment of FPV-vaccine effect than HI test especially in cats with negative or low HI titer. SN titer was 1:32, FPV-protective threshold, or higher in all cats with HI titers of ≥1:20, suggesting it may be appropriate to set protective HI threshold at 1:20.

Objectives: Feline gingivostomatitis (FGS) is a painful chronic inflammatory disease of the oral cavity. The purpose of this study was to examine the frequency of detection of certain common feline bacteria and viruses to determine any potential associations with FGS., Methods: A multicentre case-control study design was conducted. In total, 72 control cats and 32 cats with FGS were included in the study. Oral swabs were cultured for bacterial identification and a PCR assay was carried out to examine the infection of feline calicivirus (FCV), feline herpesvirus-1 (FHV-1), Chlamydia felis, Mycoplasma felis and Bordetella bronchiseptica., Results: There was a significant difference in age distribution between the control and the FGS group. Based on a PCR assay, the positive rate of FCV was significantly higher in FGS cats than control animals. For other infectious pathogens, including FHV-1, C felis and M felis, there was no significant difference. Bacterial culture of oral swabs revealed that Pasteurella multocida was most frequently detected, but the detection rate was significantly lower in FGS cats. In FGS cats, the incidence of Enterococcus faecalis and anaerobic bacteria were more frequently isolated than in control cats., Conclusions and Relevance: This study indicates that the positive rate of FCV was significantly higher in cats with FGS, and the microflora of the oral cavity of cats with FGS might be disrupted, although additional studies are required to compare the oral microbiome in cats of a variety of ages.

Eimeria tenella is an intracellular apicomplexan parasite, which infects cecal epithelial cells from chickens and causes hemorrhagic diarrhea and eventual death. We have previously reported the comparative RNA sequence analysis of the E. tenella sporozoite stage between virulent and precocious strains and showed that the expression of several genes involved in mitochondrial electron transport chain (ETC), such as type II NADH dehydrogenase (NDH-2), complex II (succinate:quinone oxidoreductase), malate:quinone oxidoreductase (MQO), and glycerol-3-phosphate dehydrogenase (G3PDH), were upregulated in virulent strain. To study E. tenella mitochondrial ETC in detail, we developed a reproducible method for preparation of mitochondria-rich fraction from sporozoites, which maintained high specific activities of dehydrogenases, such as NDH-2 followed by G3PDH, MQO, complex II, and dihydroorotate dehydrogenase (DHODH). Of particular importance, we showed that E. tenella sporozoite mitochondria possess an intrinsic ability to perform fumarate respiration (via complex II) in addition to the classical oxygen respiration (via complexes III and IV). Further analysis by high-resolution clear native electrophoresis, activity staining, and nano-liquid chromatography tandem-mass spectrometry (nano-LC-MS/MS) provided evidence of a mitochondrial complex II-III-IV supercomplex. Our analysis suggests that complex II from E. tenella has biochemical features distinct to known orthologues and is a potential target for the development of new anticoccidian drugs.

Total body irradiation (TBI) using helical tomotherapy (HT) has advantages over the standard linear accelerator-based approach to the conditioning regimen for hematopoietic cell transplantation. However, the radiation field has to be divided into two independent irradiation plans to deliver a homogeneous dose to the whole body. A clinical target volume near the skin increases the skin surface dose; therefore, high- or low-dose regions arise depending on the set-up position accuracy because the two radiation fields are somewhat overlapped or separated. We aimed to determine an adequate treatment planning method robust to the set-up accuracy for the field joint dose distribution using HT-TBI. We calculated treatment plans reducing target volumes at the interface between the upper and lower body irradiations and evaluated these joint dose distributions via simulation and experimental studies. Target volumes used for the optimization calculation were reduced by 0, 0.5, 1.0, 2.0, 2.5, and 3.0 cm from the boundary surface on the upper and lower sides. Combined dose distributions with set-up error simulated by modifying coordinate positions were investigated to find the optimal planning method. In the ideal set-up position, the target volume without a gap area caused field junctional doses of up to approximately 200%; therefore, target volumes reduced by 2.0-3.0 cm could suppress the maximum dose to within 150%. However, with set-up error, high-dose areas exceeding 150% and low-dose areas below 100% were found with 2.0 and 3.0 cm target volume reduction. Using the dynamic jaw (DJ) system, dose deviations caused by set-up error reached approximately 20%, which is not suitable for HT-TBI. Moreover, these dose distributions can be easily adjusted when combined with the intensity modulation technique for field boundary regions. The results of a simulation and experimental study using a film dosimetry were almost identical, which indicated that reducing the target volume at the field boundary surface by 2.5 cm produces the most appropriate target definition., (Copyright © 2018 American Association of Medical Dosimetrists. Published by Elsevier Inc. All rights reserved.)

Background: Palliative therapeutic strategies in esophageal squamous cell carcinoma (ESCC) patients with dysphagia remain controversial. Only few studies have assessed therapeutic effect factors related to improvement in dysphagia score and nutrition-support-free survival (NSFS)., Objective: The present study assessed the efficacy and therapeutic effect factors related to the use of palliative radiotherapy (RT) and chemoradiotherapy (CRT) in ESCC patients with dysphagia., Methods: We retrospectively evaluated 70 patients with stage IVA/B ESCC. Patients received RT of 30 Gy in 10 fractions or concurrent CRT using 5-fluorouracil plus cisplatin of 40 Gy in 20 fractions. The change in the dysphagia score from before to after treatment was assessed, and NSFS was evaluated., Results: The median follow-up duration was 6 months (range 1-41 months). The overall rate of improvement in the dysphagia score was 60%. The median NSFS was 7.5 months. Craniocaudal tumor length < 6 cm, tumor circumference < 3/4, and CRT of 40 Gy in 20 fractions were associated with a significant improvement in the dysphagia score (p = 0.0036, p = 0.0069, and p = 0.03, respectively). NSFS was significantly longer with CRT than with RT (p = 0.048)., Conclusion: Palliative RT and CRT are effective treatment options for ESCC patients with dysphagia. Craniocaudal tumor length < 6 cm, tumor circumference < 3/4, and CRT of 40 Gy in 20 fractions may improve dysphagia. CRT of 40 Gy in 20 fractions may improve NSFS.

We report a case of pelvic metastasis of rectal cancer that developed 10 years after curative resection. An 81-year-old woman underwent intersphincteric resection for lower rectal cancer 10 years previously. The tumor was pathologically diagnosed as T2N0M0, Stage Ⅰ. Nine years after the curative resection, serum carcinoembryonic antigen(CEA)levels were slightly elevated, but no recurrence was found on computed tomography(CT). Eleven months after CT, serum CEA levels elevated to 15.9 ng/mL. Pelvic metastasis in the piriformis muscle was detected on positron emission tomography(PET)-CT. Following CT-guided biopsy, she was pathologically diagnosed with metastatic rectal cancer. Radiotherapy (60 Gy/30 Fractions) was administered. Ten months after radiotherapy, PET-CT revealed no relapse in the pelvis with lung metastases.

Recent research suggests that raccoons (Procyon lotor) can transmit several important pathogens affecting humans, including protozoans. In Japan, the number of wild raccoons has increased since they were first introduced more than 50 years ago. Here, we report the first survey of Cryptosporidium infection using fecal swabs of raccoons captured in Osaka, Japan. Of 116 raccoons examined by PCR targeting of the Cryptosporidium 18S rRNA gene, 7 (6.03%; 2 adults and 5 young animals) were positive, and the isolates were identified as Cryptosporidium skunk genotype (subtype XVIa) and C. parvum based on sequence and phylogenetic analyses. Both species and the genotype are zoonotic; thus, our results suggest that raccoons could transmit Cryptosporidium infections to humans in Japan.

Stereotactic body radiation therapy (SBRT) allows the targeting of high doses of radiation with steep dose gradients. Vertebral fracture is increasingly recognized as an adverse event after SBRT; however, no sacral fractures have been observed after SBRT. We report an extremely rare case of sacral insufficiency fracture after SBRT.

[This corrects the article DOI: 10.1371/journal.pone.0199117.].

Due to recent advancements in diagnostic techniques, the incidence of multiple primary cancer has increased; however, synchronous cancer of the anal canal and cervix (SCACC) is rare, and no previous studies have investigated the treatment of this disease. The present study reports a case in which intensity-modulated radiotherapy (IMRT) was used to treat a 64-year-old female with SCACC, inguinal lymphadenopathy and anal pain. The patient was diagnosed with cT3N3M0 stage IIIb anal canal squamous cell carcinoma and cT1b1N0M0 stage Ib1 cervical squamous cell carcinoma, based on biopsy and imaging study data. According to the definitive treatment for advanced-stage anal canal cancer, outpatient treatment with chemoradiotherapy (CRT) using S-1 for SCACC was recommended, as the patient did not want to undergo resection of the anus. Considering the lymph node regions involved in SCACC and the necessary doses, the treatment plan was as follows: Whole pelvis and inguinal lymph node region radiation (36 Gy/20 fractions); a first booster radiation dose (9 Gy/5 fractions) for the whole pelvis; and a second booster radiation dose (14.4 Gy/8 fractions) for the primary lesions. The patient was prescribed S-1 at a dose of 60 mg/m 2 /day twice daily on days 1-14 and 29-42. The patient experienced grade 2 diarrhea and anal mucositis, but CRT was completed without discontinuation and hospitalization. The patient exhibited a complete response and remained disease-free without any treatment-associated complications at the 6-month follow-up. In conclusion, SCACC was successfully treated with IMRT in the present case. It is important to determine the treatment strategy for synchronous cancer types, taking into consideration the tumor stage, tumor location and patient situation.

Background: For patients with postoperative lymph node (LN) recurrent esophageal cancer, the appropriate irradiation field in chemoradiotherapy (CRT) remains controversial. We assessed the clinical outcomes and prognostic factors related to involved-field CRT for postoperative solitary LN recurrence of esophageal squamous cell carcinoma (ESCC)., Methods: We retrospectively evaluated 21 patients who had received curative resection, with LN recurrence of ESCC. Patients received CRT using 5-fluorouracil plus cisplatin or docetaxel, prescribed at 60 Gy in 30 fractions. We evaluated the pattern of failure, toxicities, survivals, and prognostic factors. We defined elective nodal failure (ENF) as recurrence in a regional LN without involved-field failure., Results: The median follow-up duration was 32 months (range, 4-106 months). Nine patients experienced failure-4 (19%) within involved-field and 5 (24%) with distant metastasis. No patients had ENF. We observed no severe toxicities. The 2-year overall survival (OS) rate was 78%. In the univariate analysis of OS, two factors, the maximal diameter of the metastatic LN < 25 mm and the absence of serum p53 antibodies (s-p53-Abs), were associated with a significantly better prognosis (p = 0.025 and p = 0.01, respectively)., Conclusions: Involved-field CRT for postoperative solitary LN recurrence of ESCC did not cause ENF and was without severe toxicities. Two factors, a length of the metastatic LN < 25 mm and the absence of s-p53-Abs may improve the treatment outcome. Involved-field CRT is a treatment option worthy of consideration for postoperative solitary LN recurrence of ESCC.

Objective: In the treatment of head-neck cancer, parotid-sparing intensity-modulated radiotherapy (IMRT) could reduce the incidence of xerostomia. When the parotid glands cannot be sufficiently spared because of the widespread tumor, contralateral superficial lobe parotid-sparing (CSLPS)-IMRT could be used to reduce marginal recurrence, however the success of this approach remains to be determined. The primary purpose of this study was to compare the incidence of xerostomia between three-dimensional conformal radiotherapy (3D-CRT) and CSLPS-IMRT for oropharyngeal and hypopharyngeal cancer. In a second aim, we also compared the clinical efficacy of 3D-CRT and CSLPS-IMRT., Methods: We retrospectively reviewed the medical records of locally advanced oropharyngeal and hypopharyngeal cancer patients who were treated with definitive concurrent chemoradiotherapy between June 2007 and April 2014. We estimated the average mean dose delivered to the parotid glands, the incidence of xerostomia≥Grade 2, patterns of failure, and survival outcomes., Results: Seventeen patients received 3D-CRT and 21 received CSLPS-IMRT. The average mean dose delivered to the superficial lobe of the contralateral parotid gland was 45.3Gy and 26.6Gy (p<0.001), and the incidence of xerostomia≥Grade 2 following treatment was 75% and 26% at 12 months (p=0.012) and 67% and 18% at 24 months (p=0.018) in the 3D-CRT and CSLPS-IMRT groups, respectively. Patterns of failure did not differ between the two groups. The 2-year progression-free survival was 59% and 62% (p=0.73), and the 2-year overall survival rate was 71% and 71% in the 3D-CRT and CSLPS-IMRT groups, respectively (p=0.76)., Conclusion: Incidence of xerostomia was significantly lower in patients receiving CSLPS-IMRT compared with 3D-CRT, while clinical efficacy did not differ between two treatment strategies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A myeloablative regimen that includes total-body irradiation (TBI) before hematopoietic stem cell transplantation results in higher patient survival rates than achieved with regimens without TBI. The TBI protocol, however, varies between institutions. In October 2015, the Japanese Radiation Oncology Study Group initiated a national survey of myeloablative TBI (covering 2010-2014). Among the 186 Japanese institutions performing TBI, 90 (48%) responded. The 82 institutions that had performed myeloablative TBI during this period treated 2698 patients with malignant disease [leukemia (2082 patients, 77.2%), malignant lymphoma (378, 14%)] and 37 with non-malignant disease [severe aplastic anemia (20, 54%), inborn errors of metabolism (5, 14%)]. A linear accelerator was used at all institutions. The institutions were divided into 41 large and 41 small institutions based on the median number of patients. The long source-surface distance technique was the method of choice in the 34 institutions (82.9%) and the moving-couch technique in the 7 (17.1%) in the large institutions. The schedules most routinely used by the participating institutions consisted of 12 Gy/6 fractions/3 days (26 institutions, 63.5%) in the large institutions. The dose rate varied from 5 to 26 cGy/min. The lungs and lenses were routinely shielded in 23 large institutions (56.1%), and only the lungs in 9 large institutions (21.9%). At lung-shielding institutions, the most frequent maximum acceptable total dose for the lungs was 8 Gy (19 institutions, 27.5%). Our results reveal considerable differences in the TBI methods used by Japanese institutions and thus the challenges in designing multicenter randomized trials based on TBI.

The effects of the high-dose ionizing radiation used in radiotherapy have been thoroughly demonstrated in vitro and in vivo. However, the effects of low-dose ionizing radiation (LDIR) such as computed tomography-guided biopsies and X-ray fluoroscopy on skin cells remain controversial. This study investigated the molecular effects of LDIR on the human primary keratinocytes (HPKs) and U937 cells, monocytes-like cell lines. These cells were exposed to 0.1 Gray (Gy) X-ray as LDIR. The modulation of transcription was assessed using a cDNA array, and the protein expression after LDIR exposure was investigated using isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis at 24 hours. These effects were confirmed by immunoblotting analysis. The direct effects of LDIR on the U937 cells and HPKs and the bystander effects of irradiated HPKs on U937 cells were also investigated. LDIR downregulated c-Myc in both U937 cells and HPKs, and upregulated the p21WAF1/CIP1 protein expression in U937 cells along with the activation of TGFβ and protein phosphatase 2A (PP2A). In HPKs, LDIR downregulated the mTOR signaling with repression of S6 and 4EBP1 activation. Similar changes were observed as bystander effects of LDIR. Our findings suggest that LDIR inhibits protein synthesis and induces the cytokines activation associated with inflammation via direct and bystander effects, which might recapitulate the effects of LDIR in inflammated skin structures., Competing Interests: The authors have declared that no competing interests exist.

The difficulty of measuring output factor (OPF) in a small field has been frequently discussed in recent publications. This study is aimed to determine the OPF in a small field using 10-MV photon beam and stereotactic conical collimator (cone). The OPF was measured by two diode detectors (SFD, EDGE detector) and one micro-ion chamber (PinPoint 3D chamber) in a water phantom. A Monte Carlo simulation using simplified detector model was performed to obtain the correction factor for the detector measurements. About 12% OPF difference was observed in the measurement at the smallest field (7.5 mm diameter) for EDGE detector and PinPoint 3D chamber. By applying the Monte Carlo-based correction factor to the measurement, the maximum discrepancy among the three detectors was reduced to within 3%. The results indicate that determination of OPF in a small field should be carefully performed. Especially, detector choice and appropriate correction factor application are very important in this regard.

This study aimed to verify the validity of generating treatment plans for volumetric arc therapy (VMAT) for prostate cancer using magnetic resonance (MR) imaging with a dose calculation algorithm in Acuros XB (Eclipse version 13.6; Varian Medical Systems, Palo Alto, CA, USA) based on deterministically solving the linear Boltzmann transport equations. Four different classes were applied to prostate MR images: MR W (all water equivalent); MR W+B (water and bone); MR S+B (soft tissue and bone); and MR S+B+G (soft tissue, bone, and rectal gas). Each of these regions was assigned a mass density for calculating doses. The assigned mass-density values were then altered in three ways. Using initial planning and optimization parameters, MR-based VMAT plans were generated and compared with corresponding forward-calculated computed tomography-based plans for doses to the target volumes and organs at risk using dose-volume histograms and γ analyses. In the MR W plans, the mean doses for TVs were overestimated by approximately 1.3%. The MR W+B plans revealed reduced differences within 0.5%. Further segmentation (MR S+B ) did not result in substantial improvement. Dose deviations affected by the changes in the mass densities assigned to soft tissue were as small as approximately 1.0%, whereas larger deviations were revealed in bone and rectal gas, especially those with > 5% error. Assignment of accurate mass-density values acquired from MR images is needed for MR-based radiation treatment planning. Multiple MR sequences should be acquired for segmentation and mass-density conversion purposes. Segmented MR-based VMAT planning is feasible with a density assignment method using Acuros XB.

The link between factor (F)VIII and FX is essential for optimum activity of the tenase complex. The interactive site(s) in FVIII for FX remains to be completely clarified, however. We investigated the FVIII A2 domain-FX association that was speculated from inhibitory mechanism(s) by an anti-A2 autoantibody. SDS-PAGE demonstrated that the purified inhibitor IgG recognizing residues 373-562 blocked FXa cleavage at Arg372 in FVIII, and surface-plasmon resonance (SPR)-based assays showed that intact A2 subunit directly bound to FX ( K d ; 63 nM). The FVIII structure model indicated possible FX-binding site(s) in residues 400-429 in A2. One peptide corresponding to residues 400-409 competitively inhibited both the A2-FX binding and FVIIIa/FIXa-dependent FXa generation. Covalent cross-linking was observed between this peptide and FX following reaction with EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide) using SDS-PAGE. K408 and S409 were not evident in N-terminal sequence analysis of the cross-linked product, suggesting that two residues participated in cross-link formation. SPR-based assays using recombinant FVIII mutants with one or both residues substituted to alanine demonstrated that K408A and K408A/S409A had approximately fourfold high K d values of wild-type (WT-)FVIII. FXa cleavages at Arg372 in both mutants were significantly delayed, suggesting a contribution of K408 for FXa cleavage at Arg372. Furthermore, FXa generation assays with these mutants demonstrated that the K m values were 1.4- to 1.7-fold greater, and overall catalytic efficiency ( k cat / K m ) was 0.49- to 0.89-fold lower than with WT-FVIII, suggesting a significant contribution of K408 for FVIII-FX interaction in tenase assembly. We concluded that the K408 in the A2 domain provided an interactive-site for FX., Competing Interests: The authors declare that they have no conflict of interest., (Schattauer GmbH Stuttgart.)

Background: The therapeutic strategies and prognostic risk factors in patients with lymph node (LN) recurrence of esophageal cancer remain controversial. We assessed clinical outcomes and prognostic factors related to the use of chemoradiotherapy (CRT) for LN recurrence of esophageal squamous cell carcinoma (ESCC) after curative resection., Methods: We retrospectively evaluated survival and prognostic factors in 57 patients with LN recurrence of ESCC after curative resection. Patients received CRT using 5-fluorouracil plus cisplatin (FP) or docetaxel. Radiotherapy was delivered at 2 Gy (total dose, 60-66 Gy; median, 60 Gy)., Results: The median follow-up duration was 24 (range, 3-116) months. The overall survival (OS) rates at 2, 3 and 5 years were 43.7%, 36.9% and 27.6%, respectively. In the univariate analysis of OS, treatment with FP, a single LN recurrence, and a single regional recurrence were associated with a significantly better prognosis (P = 0.04, P = 0.027 and P = 0.0001, respectively). In the multivariate analysis, the combination chemotherapy regimen [hazard ratio (HR), 2.50; 95% confidence interval (CI), 1.23-5.07] and the number of the regional LNs with recurrence (HR, 5.76; 95% CI, 1.22-27.12) were independent prognostic factors., Conclusion: Approximately 28% of ESCC patients with LN recurrence after curative resection could achieve long-term survival with CRT. Treatment with FP or patients with a single regional recurrence might improve the treatment outcome., (© The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com)

OBJECTIVE To evaluate outcome of limb fracture repair in rabbits. DESIGN Retrospective case series. ANIMALS 139 client-owned rabbits with limb fractures treated between 2007 and 2015. PROCEDURES Medical records were reviewed for information on fracture location, fracture treatment, and time to fracture healing. RESULTS 25 rabbits had fractures involving the distal aspects of the limbs (ie, metacarpal or metatarsal bones, phalanges, and calcaneus or talus). Fractures were treated in 23 of these 25 rabbits (external coaptation, n = 17; external skeletal fixation, 4; and intramedullary pinning, 2) and healed in all 23, with a median healing time of 28 days (range, 20 to 45 days). One hundred ten rabbits had long bone fractures, and fractures were treated in 100 of the 110 (external skeletal fixation, n = 89; bone plating, 1; intramedullary pinning, 3; and external coaptation, 7). The percentage of fractures that healed was significantly lower for open (14/18) than for closed (26/26) tibial fractures and was significantly lower for femoral (19/26) and treated humeral (4/6) fractures than for radial (23/24) or closed tibial (26/26) fractures. Micro-CT was used to assess fracture realignment during external skeletal fixator application and to evaluate fracture healing. CONCLUSIONS AND CLINICAL RELEVANCE The prognosis for rabbits with limb fractures was good, with fractures healing in most rabbits following fracture repair (109/123). Micro-CT was useful in assessing fracture realignment and evaluating fracture healing.

Aim: Safety and efficacy of vortioxetine (5-20 mg/day) in Japanese patients with major depressive disorder were evaluated in two phase 3 studies consisting of a short-term, 8-week, placebo-controlled, double-blind study followed by a long-term, 52-week, open-label extension study., Methods: The primary end-point of the short-term study was change from baseline in Montgomery-Åsberg Depression Rating Scale (MADRS) total score at week 8. The primary objective of the extension study was vortioxetine's long-term safety; efficacy end-points included change in MADRS total score, Clinical Global Impression Scale (CGI)-Severity (S) score from the long-term study baseline, and CGI-Improvement (CGI-I) score over 52 weeks., Results: Of the 366 randomized patients, 338 completed the short-term study, and 119 patients continued into the extension study. Primary (analysis of covariance) and secondary (mixed model for repeated measurements) analyses in the short-term study showed numerically greater, but not statistically significant, decreases in change in MADRS total score from baseline between the vortioxetine and placebo groups at week 8. In the long-term study, 86.6% of patients reported at least one treatment-emergent adverse event, with the most common being nasopharyngitis (40.3%) and nausea (21%). MADRS total score and CGI-I and CGI-S scores improved with continued vortioxetine treatment from baseline of the open-label study to week 52., Conclusion: Vortioxetine failed to meet significance versus placebo in the primary efficacy analysis at week 8 in the short-term study. The extension trial indicated continued improvement of depressive symptoms from baseline of this study throughout the 52-week treatment period. Vortioxetine treatment was safe and well tolerated in both studies., (© 2017 Takeda Pharmaceutical Company, Ltd. Psychiatry and Clinical Neurosciences published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Psychiatry and Neurology.)

Aim: This study assessed the efficacy and safety of vortioxetine in adults with major depressive disorder., Methods: In this double-blind, placebo-controlled study, 600 patients with major depressive disorder were randomly assigned (1:1:1:1) to receive vortioxetine 5, 10, or 20 mg, or placebo once daily for 8 weeks. The primary end-point was change from baseline in Montgomery-Åsberg Depression Rating Scale (MADRS) total score at week 8, evaluated by the last-observation-carried-forward method. Secondary end-points included response (≥ 50% decrease in the MADRS total score from baseline) and remission (MADRS total score ≤ 10), Clinical Global Impression Scale-Improvement, and change from baseline in Sheehan Disability Scale. Adverse events were summarized., Results: Vortioxetine failed to show significant differences from placebo in the primary end-point. Nominally significant improvements over placebo were observed for vortioxetine doses of 10 and 20 mg when the primary end-point was evaluated using the mixed model for repeated measures as the secondary analysis, and 10 mg in secondary measures of response and patient functioning. Vortioxetine was well tolerated. Nausea, constipation, dry mouth, dizziness, and insomnia each occurred at a >twofold higher rate than placebo. Discontinuation symptom scores were comparable between all groups after 1 and 2 weeks following withdrawal of the study drug., Conclusion: While vortioxetine failed to meet significance versus placebo in the primary efficacy analysis, there was evidence of efficacy for the 10- and 20-mg doses in secondary analyses. Vortioxetine was safe and well tolerated. Additional studies appear warranted., (© 2017 The Authors. Psychiatry and Clinical Neurosciences published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Psychiatry and Neurology.)

Most of preclinical cancer studies use xenograft models established from human cell lines or patient-derived cancer cells subcutaneously implanted into the flank of immunocompromised mice. These models are often assumed to represent the original diseases and are valuable tools, at least to some extent, for understanding both the basic biology of cancer and for proof-of-concept studies of molecularly targeted therapies. However, analyzing the cellular behavior of individual components within xenografts, including tumor cells, stromal cells, immune cells, and blood vessels, is challenging. In particular, it has been difficult and urgently required to trace the whole process of heterogeneous tumor microenvironment formation mediated by various components described above. Here we demonstrate a method for monitoring this process using a window device system that we have recently developed and a subcutaneous xenograft model that accurately recapitulates the histology of human lung adenocarcinoma. Use of our imaging window device and a multiphoton laser scanning microscope provides a powerful tool for investigating tumor heterogeneity and responses to drug treatments in an in vivo live imaging system.

To investigate the utility of cerebrospinal fluid (CSF) anti-feline coronavirus (FCoV) antibody test for diagnosis of feline infectious peritonitis (FIP), the antibody titers were tested in CSF and sera from 271 FIP-suspected neurological cats. CSF antibody was detected in 28 cats, which were divided into 2 groups; 15 with CSF titer of 1:80 or lower and 13 with CSF titer of 1:640 or higher. In the latter group, reciprocal serum titer/reciprocal CSF titer was 8 or lower, which is extremely lower than normal range (256-2048), and FCoV RNA was detected in all of 11 CSF samples assayed by RT-PCR. Our findings indicate that CSF titer of 1:640 or higher may be served as a candidate for the index for diagnosing FIP.

Purpose: To analyze the detailed pattern of intraosseous failure after stereotactic body radiation therapy (SBRT) for coxal bone metastasis., Methods and Materials: Patients treated with SBRT to coxal bone metastasis were identified by retrospective chart review. The SBRT doses were 30 Gy or 35 Gy in 5 fractions. A margin of 5 to 10 mm was added to the gross tumor volume to create the clinical target volume. We evaluated the presence or absence of intraosseous recurrence using magnetic resonance imaging. Intraosseous recurrences were assessed as "in-field" or "marginal/out-of-field." In addition, we measured the distance between the center of the recurrent tumor and the nearest edge of the initial bone metastasis in cases of marginal/out-of-field recurrence., Results: Seventeen patients treated for 17 coxal bone metastases were included. Median age was 64 years (range, 48-79 years). Coxal lesions involved the ilium in 14 cases, pubis in 3, and ischium in 4 (3 lesions crossed over multiple regions). Patients most commonly had renal cell carcinoma (29.4%), followed by lung, hepatic cell, and colorectal cancers (23.5%, 11.8%, and 11.8%, respectively). Median follow-up after SBRT was 13 months (range, 2-44 months). Among all 17 cases, 7 cases developed 8 intraosseous recurrences, including in-field recurrence in 1 case and marginal/out-of-field recurrences in 7 cases. Median time to intraosseous recurrence was 10 months (range, 2-35 months). Among 7 cases with marginal/out-of-field recurrence, mean distance to the center of the recurrent tumor from the nearest edge of the initial bone metastasis was 34 mm (range, 15-55 mm)., Conclusions: Most recurrences were observed out-of-field in the same coxal bone. These results suggest that defining the optimal clinical target volume in SBRT for coxal bone metastasis to obtain sufficient local tumor control is difficult., (Copyright © 2017 Elsevier Inc. All rights reserved.)

From a total of 72 commercial layer and pullet farms that were monitored in the eastern Japan area, 4 farms had mild to severe respiratory disease accompanied by decreased feed intake and drop in egg production. Microbiological analysis showed that 3 of the 4 farms, particularly from Fukushima, Tochigi, and Ibaraki prefectures, were positive for Ornithobacterum rhinotracheale (ORT). Out of 65 birds examined, ORT was isolated in 21 birds (32.31%). All isolates were Gram-negative pleomorphic rods with a colony size of 0.05 mm, translucent with grayish coloration, and with butyric smell after 48 h of incubation in 10% chicken blood agar at 37°C under microaerophilic conditions. All isolates reacted positively in the p-nitrophenyl-β-d-galactopyranoside test within 3 h and were positive in cytochrome oxidase tests with an API 20NE identification system biocode of 0-0-2-0-0-0-4. An agar gel precipitation test showed that all isolates were serotype-A. All strains were positive in PCR by yielding a 784 bp amplicon of the 16S rRNA gene. All strains were resistant to amikacin, colistin, gentamicin, kanamycin, neomycin, polymyxin b, streptomycin, and sulfamethoxazole trimethoprim and susceptible to amoxicillin clavulanic acid, ampicillin, doxycycline, spectinomycin, and tetracycline. This study is the first characterization of ORT from commercial layer chickens in eastern Japan., (© 2017 Poultry Science Association Inc.)

Thus far, Entamoeba species have been classified based on morphology such as the number of nuclei in mature cysts and their hosts. Using recently developed molecular tools, ruminant Entamoeba spp. are currently classified into four species/genotypes: E. bovis and Entamoeba ribosomal lineages (RL) 1, 2, and 4. However, the distribution or pathogenicity of ruminant Entamoeba has not been well documented. In the present study, we examined a total of 25 fecal and seven environmental samples collected from six farms in Japan from 2016 to 2017 by the floatation method and PCR and sequencing analyses. Consequently, we detected Entamoeba cysts in 18 of 25 cattle samples and four of the seven environmental samples, including soil and drinking water, by microscopic examinations. In sequential examinations, Entamoeba-positive cattle were found to shed cysts without any clinical symptoms for more than 8 months. By PCR for molecular identification, isolates in ten cattle and one soil sample were successfully sequenced and formed a cluster of E. bovis, which was separated from those of other Entamoeba species/genotypes such as RL1-4 in phylogenetic analysis. To our knowledge, this is the first report about E. bovis in Japan, and our results may implicate that E. bovis is not pathogenic.

Bovine intranuclear coccidiosis is caused by the protozoans Eimeria alabamensis and Cyclospora spp. Here, we characterized the disease and genetically identified the causative species in Japanese black calves with chronic and refractory watery diarrhea. Histologic examinations revealed atrophy of the jejunal villi and numerous parasites in the nucleus of epithelial cells in the jejunum. Based on molecular analyses using 18S ribosomal RNA gene-specific primers that we designed, the parasites were found to be formed in the same cluster as Eimeria subspherica in the phylogenetic tree, which was separated from those of other related Eimeria spp. These results constitute the first report of E. subspherica as a cause of bovine intranuclear coccidiosis.

Raccoons (Procyon lotor) are found worldwide. They are frequently seen in crowded inner cities as well as in forests or wooded areas, often living in proximity to humans and their pets. We examined sera from 100 wild raccoons in Japan for antibodies to six canine viruses with veterinary significance to assess their potential as reservoirs. We also aimed to understand the distribution of potentially infected wildlife. We found that 7% of samples were seropositive for canine distemper virus (CDV), 10% for canine parvovirus type 2, 2% for canine adenovirus type 1, 6% for canine adenovirus type 2, and 7% for canine coronavirus. No samples were found to be seropositive for canine parainfluenza virus. Seropositivity rates for canine distemper virus and canine parvovirus type 2 were significantly different between areas, and younger raccoons (<1 yr old) were more frequently seropositive than older raccoons. Because raccoons belong to the suborder Caniformia, similar to dogs (Canis lupus familiaris), our results suggest that they can act as reservoirs for some of these important canine viruses and might be involved in viral transmission. Further study should include isolation and analysis of canine viruses in wild raccoons from a wider area.

The incidence of lung adenocarcinoma has been increasing recently in smokers. The molecular target therapy has been developed for lung adenocarcinoma patients harboring EGFR gene mutation. However, the treatment modalities for patients without mutation are currently limited. Thus, analysis of EGFR gene mutation status at early stage is important strategy to classify the patients for improving treatments and prognosis efficiently. This study aimed to identify microRNA (miRNA) signature in relation to mutation status in EGFR gene in early stage of lung adenocarcinoma male patients with smoking history. MiRNA profiles were assessed by microarray in paired plasma and tissue pooled from 10 EGFR wild type (EGFR-wt) and 10 EGFR mutated (EGFR-mut) patients. Expressions of selected miRNAs were verified further by real-time qRT-PCR in 83 plasma samples consisting of 55 EGFR-wt patients and 28 EGFR-mut patients and their correlation with clinicopathological parameters and EGFR gene mutation status were evaluated. We found that seven miRNAs (miR-16-5p, miR-23a-3p, miR-103a-3p, miR122-5p, miR-223-3p, miR-346 and miR-451a) were differentially expressed in stage I and stage I+II. Especially, miR-23a-3p was only miRNA shown higher expression in EGFR-wt patients than EGFR-mut patients. Thus, our findings could be useful non-invasive biomarkers to differentiate mutation status in EGFR gene in smoker lung adenocarcinoma male patients., Competing Interests: CONFLICTS OF INTEREST The authors disclose no potential conflicts of interest.

Entamoeba suis and Entamoeba polecki subtypes (ST) 1 and 3 have recently been implicated in disease outbreaks in pigs. However, the distributions of these parasites in Japan and the potential sources of infection on farms still remain unclear. Here, we examined a farm of fattening/growing pigs with abnormal feces in Kagoshima Prefecture, Japan, and found the presence of parasites in the farm environment. Examination of intestinal tissues from pigs presenting with ulcerative colitis revealed a large number of trophozoites that had invaded the lesions. We identified single and mixed infections of E. suis and E. polecki ST1 and ST3 in paraffin sections or fecal samples from affected pigs. Two subtypes of Entamoeba were identified using four primer sets by PCR and sequencing. The parasites were detected in moist soil samples obtained around the drinking water source or puddles, implicating transmission of cysts via contaminated soils. Additionally, we found evidence of Entamoeba spp. and coinfections in surveyed pigs without any diarrhea at two neighboring farms. Our results establish methods for successfully identification of parasites, including cases in which multiple infections are present.

It has been proposed that a subpopulation of tumour cells with stem cell-like characteristics, known as cancer stem cells (CSCs), drives tumour initiation and generates tumour heterogeneity, thus leading to cancer metastasis, recurrence, and drug resistance. Although there has been substantial progress in CSC research into many solid tumour types, an understanding of the biology of CSCs in lung cancer remains elusive, mainly because of their heterogeneous origins and high plasticity. Here, we demonstrate that engineered lung cancer cells derived from normal human airway basal epithelial cells possessed CSC-like characteristics in terms of multilineage differentiation potential and strong tumour-initiating ability. Moreover, we established an in vitro 3D culture system that allowed the in vivo differentiation process of the CSC-like cells to be recapitulated. This engineered CSC model provides valuable opportunities for studying the biology of CSCs and for exploring and evaluating novel therapeutic approaches and targets in lung CSCs.

In the production and management of beef and dairy cattle, controlling diarrhea is one of the important concerns. Pathogenic agents of the disease, protozoan parasites including Cryptosporidium spp., are difficult to control, making prevention, diagnoses, and treatment of diarrhea. In the present study, we investigated a farm with a history of calf deaths over a period of 10 years in order to determine the cause of disease and to clarify the detailed distribution of the pathogens. In four examined calves that were reared in calf pens, all were positive with Cryptosporidium and/or Giardia, while the other breeding stock and adult cattle were negative. Molecular analyses revealed that the isolates from calves were C. parvum subtype IIaA15G2R1 as a zoonotic and G. intestinalis assemblage E. Other pathogenic bacteria and diarrhea-causing viruses were not detected. After treating the calf pens with boiling water and milk of lime (Ca[OH]2), oocysts of C. parvum and cysts of G. intestinalis were not found and no additional calves died. This is the first report to describe the mixed infection of both parasites in Japan.

Human RAD17 acts as an activator of checkpoint signals in response to DNA damage. Here we evaluated the association of hRAD17 Leu546Arg (rs1045051), a missense single nucleotide polymorphism, with the risk of colorectal cancer (CRC) in relation to smoking and alcohol consumption habits in 212 CRC patients and 1,142 cancer-free controls in a case-control study conducted in Japan. The results showed that the hRAD17 Leu/Arg genotype compared to the Leu/Leu genotypes was significantly associated with the protective effect on CRC risk with the adjusted odds ratio (OR) of 0.68 [95% confidence interval (CI): 0.49-0.95, p=0.024], and the males with the Arg/Arg genotype had a greater risk of CRC compared to those with the Leu/Leu and Leu/Arg genotypes (OR=1.87, 95%CI 1.03-3.40, p=0.04). In stratified studies, the protective effect of the Leu/Arg genotype on CRC risk was markedly higher in the light smokers (< 20 pack years) (OR=0.61, 95%CI 0.40-0.94, p=0.024) and the rectal cancer patients (OR=0.49, 95%CI 0.31-0.78, p=0.003). The risk of the Arg/Arg genotype was associated with heavy smoking (≥ 20 pack-years) (OR=2.24, 95%CI 1.09-4.61, p=0.03). These findings suggest that the genetic variant of hRAD17 Leu546Arg polymorphism has a significant effect on CRC susceptibility in Japanese., Competing Interests: No potential conflict of interest relevant to this article was reported.

Objective: The present study aimed to clarify gross tumor volume (GTV) contouring accuracy at the diaphragm boundary using respiratory-gated PET/CT., Methods: The lung/diaphragm boundary was simulated using a phantom containing 18 F solution (10.6 kBq/mL). Tumors were simulated using spheres (diameter, 11-38 mm) containing 18 F and located at the positions of the lungs and liver. The tumor background ratios (TBR) were 2, 4, and 8. The phantom was moved from the superior to inferior direction with a 20-mm motion displacement at 3.6 s intervals. The recovery coefficient (RC), volume RC (VRC), and standardized uptake value (SUV) threshold were calculated using stationary, non-gated (3D), and gated (4D) PET/CT., Results: In lung cancer simulation, RC and VRC in 3D PET images were, respectively, underestimated and overestimated in smaller tumors, whereas both improved in 4D PET images regardless of tumor size and TBR. The optimal SUV threshold was about 30% in 4D PET images. In liver cancer simulation, RC and VRC were, respectively, underestimated and overestimated in smaller tumors, and when the TBR was lower, but both improved in 4D PET images when tumors were >17 mm and the TBR was >4. The optimal SUV threshold tended to depend on the TBR., Conclusions: The contouring accuracy of GTV was improved by considering TBR and using an optimal SUV threshold acquired from 4D PET images.

To investigate whether kokuvirus is present in Japanese dogs, we examined the fecal samples obtained from 94 diarrheal household dogs and 50 clinically healthy kenneled dogs by RT-PCR. The gene was detected in 37.2% and 48.0% in the former and the latter, respectively, suggesting that canine kobuvirus (CaKoV) is circulating among Japanese dogs. From the result of the latter, however, CaKoV may not be a primary pathogen. Furthermore, all gene-positive dogs were purebreds aged four months or younger. This finding suggests that CaKoV endemic is confined in multi-dog environments, and the dogs have a strong age-dependent resistance to CaKoV.

Recent data from The Cancer Genome Atlas analysis have revealed that Aurora kinase A (AURKA) amplification and overexpression characterize a distinct subset of human tumors across multiple cancer types. Although elevated expression of AURKA has been shown to induce oncogenic phenotypes in cells in vitro, findings from transgenic mouse models of Aurora-A overexpression in mammary glands have been distinct depending on the models generated. In the present study, we report that prolonged overexpression of AURKA transgene in mammary epithelium driven by ovine β-lactoglobulin promoter, activated through multiple pregnancy and lactation cycles, results in the development of mammary adenocarcinomas with alterations in cancer-relevant genes and epithelial-to-mesenchymal transition. The tumor incidence was 38.9% (7/18) in Aurora-A transgenic mice at 16 months of age following 4-5 pregnancy cycles. Aurora-A overexpression in the tumor tissues accompanied activation of Akt, elevation of Cyclin D1, Tpx2 and Plk1 along with downregulation of ERα and p53 proteins, albeit at varying levels. Microarray comparative genomic hybridization (CGH) analyses of transgenic mouse mammary adenocarcinomas revealed copy gain of Glp1r and losses of Ercc5, Pten and Tcf7l2 loci. Review of human breast tumor transcriptomic data sets showed association of these genes at varying levels with Aurora-A gain of function alterations. Whole exome sequencing of the mouse tumors also identified gene mutations detected in Aurora-A overexpressing human breast cancers. Our findings demonstrate that prolonged overexpression of Aurora-A can be a driver somatic genetic event in mammary adenocarcinomas associated with deregulated tumor-relevant pathways in the Aurora-A subset of human breast cancer., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Aurora kinases play critical roles in regulating spindle assembly, chromosome segregation, and cytokinesis to ensure faithful segregation of chromosomes during mitotic cell division cycle. Molecular and cell biological studies have revealed that Aurora kinases, at physiological levels, orchestrate complex sequential cellular processes at distinct subcellular locations through functional interactions with its various substrates. Aberrant expression of Aurora kinases, on the other hand, cause defects in mitotic spindle assembly, checkpoint response activation, and chromosome segregation leading to chromosomal instability. Elevated expression of Aurora kinases correlating with chromosomal instability is frequently detected in human cancers. Recent genomic profiling of about 3000 human cancer tissue specimens to identify various oncogenic signatures in The Cancer Genome Atlas project has reported that recurrent amplification and overexpression of Aurora kinase-A characterize distinct subsets of human tumors across multiple cancer types. Besides the well-characterized canonical pathway interactions of Aurora kinases in regulating assembly of the mitotic apparatus and chromosome segregation, growing evidence also supports the notion that deregulated expression of Aurora kinases in non-canonical pathways drive transformation and genomic instability by antagonizing tumor suppressor and exacerbating oncogenic signaling through direct interactions with critical proteins. Aberrant expression of the Aurora kinases-p53 protein family signaling axes appears to be critical in the abrogation of p53 protein family mediated tumor suppressor pathways frequently deregulated during oncogenic transformation process. Recent findings reveal the existence of feedback regulatory loops in mRNA expression and protein stability of these protein families and their consequences on downstream effectors involved in diverse physiological functions, such as mitotic progression, checkpoint response pathways, as well as self-renewal and pluripotency in embryonic stem cells. While these investigations have focused on the functional consequences of Aurora kinase protein family interactions with wild-type p53 family proteins, those involving Aurora kinases and mutant p53 remain to be elucidated. This article presents a comprehensive review of studies on Aurora kinases-p53 protein family interactions along with a prospective view on the possible functional consequences of Aurora kinase-mutant p53 signaling pathways in tumor cells. Additionally, we also discuss therapeutic implications of these findings in Aurora kinases overexpressing subsets of human tumors.