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Vector-borne pathogens exist in obligate transmission cycles between vector and reservoir host species. Host and vector shifts can lead to geographic expansion of infectious agents and the emergence of new diseases in susceptible individuals. Three bacterial genospecies (Borrelia afzelii, Borrelia bavariensis, and Borrelia garinii) predominantly utilize two distinct tick species as vectors in Asia (Ixodes persulcatus) and Europe (Ixodes ricinus). Through these vectors, the bacteria can infect various vertebrate groups (e.g., rodents, birds) including humans where they cause Lyme borreliosis, the most common vector-borne disease in the Northern hemisphere. Yet, how and in which order the three Borrelia genospecies colonized each continent remains unclear including the evolutionary consequences of this geographic expansion. Here, by reconstructing the evolutionary history of 142 Eurasian isolates, we found evidence that the ancestors of each of the three genospecies probably have an Asian origin. Even so, each genospecies studied displayed a unique substructuring and evolutionary response to the colonization of Europe. The pattern of allele sharing between continents is consistent with the dispersal rate of the respective vertebrate hosts, supporting the concept that adaptation of Borrelia genospecies to the host is important for pathogen dispersal. Our results highlight that Eurasian Lyme borreliosis agents are all capable of geographic expansion with host association influencing their dispersal; further displaying the importance of host and vector association to the geographic expansion of vector-borne pathogens and potentially conditioning their capacity as emergent pathogens.

Daily dose distributions for adaptive radiotherapy (ART) using helical tomotherapy (HT) are calculated using megavoltage computed tomography (MVCT). Generally, the MVCT number is converted to mass density (MD) using an MD calibration table (MVCT-MD table). The aims of this study are to calculate the tolerance levels of the MD for ART and to evaluate the tolerance levels using clinical patient plans. These tolerance levels of MD were calculated based on the tissue maximum ratio (TMR) of 6MV flattening-filter-free (FFF) beam of HT and the effective tissue thickness data from an International Commission on Radiological Protection 110 phantom data for lung, adipose/muscle and cartilage/spongy-bone. These tolerance levels were determined by considering both the MD causing a dose error of 2% and the variation in MVCT numbers. Subsequently, the stability of the MD values was estimated with the standard deviations (SD) in the MVCT number over 6 months. The dose distribution for clinical patient plans was calculated using the MVCT-MD table with added tolerance levels. These tolerance levels were determined as MD differences causing a dose error of 2%, and were ± 0.049 g/cm3, ± 0.030 g/cm3 and ± 0.049 g/cm3 for lung, adipose/muscle and cartilage/spongy-bone, respectively. The calculated dose distribution errors using the MVCT-MD table added tolerance levels were within 2%. We proposed these tolerance levels in MD for the quality control of the MVCT-MD table.

Treatment planning for ion therapy involves the conversion of computed tomography number (CTN) into a stopping-power ratio (SPR) relative to water. The purpose of this study was to create a CTN-to-SPR calibration table using a stoichiometric CTN calibration model with a three-parameter fit model for ion therapy, and to demonstrate its effectiveness by comparing it with a conventional stoichiometric CTN calibration model.We inserted eight tissue-equivalent materials into a CTN calibration phantom and used six CT scanners at five radiotherapy institutes to scan the phantom. We compared the theoretical CTN-to-SPR calibration tables created using the three-parameter fit and conventional models to the measured CTN-to-SPR calibration table in three tissue types: lung, adipose/muscle, and cartilage/spongy bone. We validated the estimated SPR differences in all cases and in a worst-case scenario, which revealed the largest estimated SPR difference in lung tissue.For all cases, the means ± standard deviations of the estimated SPR difference for the three-parameter fit method model were -0.1 ± 1.0%, 0.3 ± 0.7%, and 2.4 ± 0.6% for the lung, adipose/muscle, and cartilage/spongy bone, respectively. For the worst-case scenario, the estimated SPR differences of the conventional and the three-parameter fit models were 2.9% and -1.4% for the lung tissue, respectively.The CTN-to-SPR calibration table of the three-parameter fit model was consistent with that of the measurement and decreased the calibration error for low-density tissues, even for the worst-case scenario.

The aim of this study was to compare the quality of dynamic wave arc (DWA) and coplanar volumetric modulated arc therapy (co-VMAT) plans for the treatment of localized prostate cancer. The planning target volume (PTV)-rectum, a section of the PTV comprising the PTV minus that of the rectum, received 78 Gy in 39 fractions as the mean dose to the PTV-rectum. The DWA and co-VMAT plans were generated for each patient using the RayStation treatment planning system for the Vero4DRT system. The PTV-rectum dose (D95%: the percent dose irradiating 95% of the volume), homogeneity index (HI), conformity index (CI), as well as doses to the bladder wall, rectum wall (V10–70 Gy: the percent volume receiving 10–70 Gy), and bilateral femoral heads of the DWA and co-VMAT plans were compared. The output monitor unit (MU) and delivery time obtained for each set of plans were also investigated. In terms of target coverage, the DWA plans provided an average D95% of 75.5 Gy, which was comparable to the co-VMAT-plan D95% of 75.2 Gy (p

Sasaki, Mizuki, Anders, Jason Lee, Nakao, Minoru (2021): Cestode fauna of murid and cricetid rodents in Hokkaido, Japan, with assignment of DNA barcodes. Species Diversity 26: 255-272, DOI: 10.12782/specdiv.26.255

This study aims to evaluate the effect of different air computed tomography (CT) numbers of the image value density table (IVDT) on the retrospective dose calculation of head-and-neck (HN) radiotherapy using TomoTherapy megavoltage CT (MVCT) images. The CT numbers of the inside and outside air and each tissue-equivalent plug of the "Cheese" phantom were obtained from TomoTherapy MVCT. Two IVDTs with different air CT numbers were created and applied to MVCT images of the HN anthropomorphic phantom and recalculated by Planned Adaptive to verify dose distribution. We defined the recalculation dose with MVCT images using both inside and outside air of the IVDT as

New data have been obtained for three representatives of Exorchis; Exorchis convictus sp. n., Exorchis oviformis and Exorchis sp., from fish in the East-Asian region. For the first time, based on combined sequences of the ITS2 rDNA region and the 28S rRNA gene, Exorchis is confirmed to belong Cryptogonimidae. Based on analysis of a mitochondrial marker (cox1), the ‘Japanese’ and ‘Russian’ haplogroups are identified for E. oviformis isolated from Silurus asotus. One specimen of E. oviformis obtained in Japan is identical to the ‘Russian’ haplotype. Haplotype patterns are also observed for metacercariae of Exorchis sp. from Tanakia lanceolata and Carassius sp. fish in Kyushu Island (Japan).

Purpose We investigated the interfractional variation in the tumor position during lung stereotactic body radiotherapy (SBRT) under expiratory-phase breath hold (BH) using cone-beam computed tomography (CBCT). Methods A total of 79 patients with lung cancer were treated with lung SBRT, wherein the Abches system under expiratory-phase BH was used to study interfractional variation. The tumors were located in the upper lobe in 31 cases, in the middle lobe in 11 cases, and in the lower lobe in 37 cases. Planning CTs were scanned under expiratory-phase BH with the Abches system. The 3-degrees-of-freedom (DOF) tumor-based setup using CBCT images under expiratory-phase BH was performed after a 6-DOF bony vertebrae-based setup using an ExacTrac X-ray system. Interfractional variation in the lung tumor position was defined as the difference in the position of the lung tumor relative to the bone anatomy in the left-right (LR), antero-posterior (AP), and craniocaudal (CC) directions represented as absolute values. Results The interfractional variation in the lung tumor position was very similar in all the lung regions, and its mean ± standard deviation values in all patients were 1.0 ± 1.1, 1.6 ± 1.9, and 1.6 ± 1.9 mm in the LR, AP, and CC directions, respectively. Further, 99.1%, 92.4%, and 92.7% of all the fractions for the interfractional tumor positional variation in the LR, AP, and CC directions were less than 5 mm, respectively. Conclusion The interfractional variation in the tumor position was small for lung cancer patients treated with the Abches system under expiratory-phase BH.

Michajlovia turdi (Yamaguti, 1939) (Digenea: Brachylaimoidea) has been found from the Japanese thrush, Turdus cardis Temminck, 1831, in Hokkaido, the northernmost island of Japan. This is a rediscovery of M. turdi in Japan after approximately 80 years from the original description of the species as Leucochloridium turdiYamaguti, 1939. Here we redescribe the morphology of M. turdi and generate DNA barcodes for the species by sequencing nuclear ribosomal RNA (18S and 28S) and mitochondrial cytochrome c oxidase subunit I (COI) genes. Our molecular phylogenetic analysis was not effective at determining the taxonomic rank of Michajlovia in Brachylaimoidea, and consequently the genus remains as incertae sedis.

To improve the homogeneity and conformity of the irradiation dose for postoperative breast cancer including regional lymph nodes, we planned Hybrid volumetric-modulated arc therapy (VMAT), which combines conventional tangential field mainly for the chest area and VMAT mainly for the supraclavicular area and marginal zone. In this study, we compared the dosimetric impact between traditional 3D conformal radiotherapy (3DCRT) and Hybrid VMAT and observed toxicities following Hybrid VMAT. A total of 70 patients indicated between October 2016 and December 2017 were included. The prescribed dose was 50 Gy/25 fractions. For the dosimetric impact, 3DCRT and Hybrid VMAT plans were compared in each patient with respect to the dosimetric parameters. Toxicities were followed using the Common Terminology Criteria for Adverse Events version 4.0. The median follow-up duration was 319 days. For the dosimetric impact, the homogeneity index (HI) and conformity index (CI) of PTV were significantly improved in the Hybrid VMAT plan compared with that in the 3DCRT plan (HI, 0.15 ± 0.07 in Hybrid VMAT vs 0.41 ± 0.19 in 3DCRT, P

Purpose In photon radiation therapy, computed tomography (CT) numbers are converted into values for mass density (MD) or relative electron density to water (RED). CT-MD or CT-RED calibration tables are relevant for human body dose calculation in an inhomogeneous medium. CT-MD or CT-RED calibration tables are influenced by patient imaging (CT scanner manufacturer, scanning parameters, and patient size), the calibration process (tissue-equivalent phantom manufacturer, and selection of tissue-equivalent material), differences between tissue-equivalent materials and standard tissues, and the dose calculation algorithm applied; however, a CT number calibration audit has not been established. The purposes of this study were to develop a postal audit phantom, and to establish a CT number calibration audit process. Methods A conventional stoichiometric calibration conducts a least square fit of the relationships between the MD, material weight, and measured CT number, using two parameters. In this study, a new stoichiometric CT number calibration scheme has been empirically established, using three parameters to harmonize the calculated CT number with the measured CT number for air and lung tissue. In addition, the suitable material set and the minimal number of materials required for stoichiometric CT number calibration were determined. The MDs and elemental weights from the International Commission on Radiological Protection Publication 110 were used as standard tissue data, to generate the CT-MD and CT-RED calibration tables. A small-sized, CT number calibration phantom was developed for a postal audit, and stoichiometric CT number calibration with the phantom was compared to the CT number calibration tables registered in the radiotherapy treatment planning systems (RTPSs) associated with five radiotherapy institutions. Results When a least square fit was performed for the stoichiometric CT number calibration with the three parameters, the calculated CT number showed better agreement with the measured CT number. We established stoichiometric CT number calibration using only two materials because the accuracy of the process was determined not by the number of used materials but by the number of elements contained. The stoichiometric CT number calibration was comparable to the tissue-substitute calibration, with a dose difference less than 1%. An outline of the CT number calibration audit was demonstrated through a multi-institutional study. Conclusions We established a new stoichiometric CT number calibration method for validating the CT number calibration tables registered in RTPSs. We also developed a CT number calibration phantom for a postal audit, which was verified by the performances of multiple CT scanners located at several institutions. The new stoichiometric CT number calibration has the advantages of being performed using only two materials, and decreasing the difference between the calculated and measured CT numbers for air and lung tissue. In the future, a postal CT number calibration audit might be achievable using a smaller phantom.

New data have been obtained for three representatives of Exorchis; Exorchis convictus sp. n., Exorchis oviformis and Exorchis sp., from fish in the East-Asian region. For the first time, based on combined sequences of the ITS2 rDNA region and the 28S rRNA gene, Exorchis is confirmed to belong Cryptogonimidae. Based on analysis of a mitochondrial marker (cox1), the ‘Japanese’ and ‘Russian’ haplogroups are identified for E. oviformis isolated from Silurus asotus. One specimen of E. oviformis obtained in Japan is identical to the ‘Russian’ haplotype. Haplotype patterns are also observed for metacercariae of Exorchis sp. from Tanakia lanceolata and Carassius sp. fish in Kyushu Island (Japan).

A nationwide fish survey was conducted in Japan to detect metacercariae of the genus Metagonimus (Trematoda: Heterophyidae). The metacercariae were subjected to DNA barcoding for molecular species identification. A phylogeny inferred from the sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) prompted us to recognize three cryptic species complexes (i.e., the M. miyatai complex, the M. takahashii complex, and the M. katsuradai complex). Each complex included one or two undescribed species. For morphological description, adult flukes of each species were raised through the experimental infections of immunosuppressed mice. We propose M. saitoi n. sp., M. kogai n. sp., M. shimazui n. sp., and M. kinoi n. sp., based on their phylogeny, morphology, biogeography, and ecology (host-parasite relationships). The originally described species, M. miyatai, was split into M. miyatai sensu stricto and M. saitoi n. sp. The former is distributed mainly in eastern Japan and uses the sweetfish (Plecoglossus altivelis) and daces (Pseudaspius hakonensis and Ps. sachalinensis) as principal second intermediate hosts, while the latter is in western Japan and its principal fish hosts are the dark chub (Nipponocypris temminckii) and the pale chub (Opsariichthys platypus). The present survey resolves a long-standing controversy on the microtaxonomy of Metagonimus in Japan since the first discovery of Metagonimus yokogawai in 1912, and shows that 10 species of Metagonimus are still distributed in Japan, although human metagonimiasis is almost eradicated.

Pulsating broodsacs of Leucochloridium sp. (Trematoda: Leucochloridiidae) were found from amber snails (Succinea lauta) in Iwate, the northern part of Honshu, Japan. A pattern with red-brown vertical stripes was characteristic of the broodsac. Very similar broodsacs were already detected from Okinawa Islands, the southern archipelago of Japan, and tentatively identified as Leucochloridium cf. passeri. A phylogenetic analysis based on DNA sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) showed that Leucochloridium sp. is different at species level from L. cf. passeri and that both species are related to Leucochloridium vogtianum from Europe. In this study the definitive identification of larval Leucochloridium sp. was impossible, but the resulting phylogeny confirmed that at least 4 species of Leucochloridium are distributed in Japan, depending on locality and climate. The DNA barcode generated in this study will be useful in detecting the adult stage of Leucochloridium sp. from birds.

The genus Rhabdias StilesHassall, 1905 comprises lung parasites of amphibians and reptiles worldwide. In Japan, 9 species have been recorded, including Rhabdias incerta Wilkie, 1930 which has been reported only in Bufo species. In this study, to assess the diversity of R. incerta, we performed molecular analyses of Rhabdias species sampled from three species/subspecies of Japanese toads namely Bufo japonicus, B. japonicus formosus, and B. torrenticola, collected in various regions of Honshu, Shikoku, and Kyushu, Japan. DNA sequence divergence was compared using mtDNA (COI) and nuclear DNA (28S) to identify possible cryptic species. Morphological analysis was performed through light microscopy and scanning electron microscopy (SEM). The results revealed that Bufo spp. serve as hosts for at least three Rhabdias species. Morphologically, most samples were identified as R. incerta but with a longer body and larger buccal cavity than originally described. Rhabdias incerta appears to be specific to the genus Bufo and is further subdivided into two or three phylogroups based on subspecies divisions and biogeography of their host. Some Rhabdias specimens collected in this study resemble R. tokyoensis Wilkie, 1930, parasitic in hosts from the order Caudata, which suggests host switching. Both molecular and morphological analyses suggested the presence of undescribed and cryptic Rhabdias species within toads collected in Japan. This study was the first to molecularly characterize Rhabdias species in Japan, including novel sequences of R. incerta and two undescribed species.

Background Few studies have assessed hydrogel spacer shrinkage during external-beam radiation therapy following brachytherapy for localized high-risk prostate cancer. This case presentation evaluated the changes in hydrogel spacer appearance by magnetic resonance imaging during external-beam radiation therapy after brachytherapy for prostate cancer and analyzed the effect of this shrinkage on the dose distribution in four cases. Case presentation In all cases, we implanted 125I sources using a modified peripheral loading pattern for seed placement. The prescribed dose for each implant was 110 Gy. After delivering the sources, a hydrogel spacer was injected. All cases underwent external-beam radiation therapy approximately 1–2 months after brachytherapy. The prescribed dose of external-beam radiation therapy was 45 Gy in 1.8-Gy fractions. Magnetic resonance imaging was performed for evaluation on the day following seed implantation (baseline), at external-beam radiation therapy planning, and during external-beam radiation therapy. The median hydrogel spacer volume was 16.2 (range 10.9–17.7) cc at baseline, 14.4 (range, 9.4–16.1) cc at external-beam radiation therapy planning, and 7.1 (range, 2.0–11.4) cc during external-beam radiation therapy. The hydrogel spacer volume during external-beam radiation therapy was significantly lower than that at external-beam radiation therapy planning. The rectum V60–80 (rectal volume receiving at least 60–80% of the prescribed dose of external-beam radiation therapy) during external-beam radiation therapy was significantly higher than that at external-beam radiation therapy planning. Conclusions The potential reduction in hydrogel spacer size during external-beam radiation therapy following brachytherapy can lead to unexpected irradiation to the rectum. This case presentation would be helpful for similar cases.

Purpose We aimed to evaluate the magnetic resonance imaging (MRI) appearance changes during stereotactic radiotherapy (SRT) for large sized brain metastases, and analyze the lesions necessitating treatment plan modification. Materials and methods A total of 23 patients (27 lesions, >2 cm in tumor diameter) underwent SRT and all lesions were evaluated the appearance changes which had the necessity of the treatment plan modification. The appearance change of tumor during SRT was evaluated using gadolinium-enhanced MRI. The reasons of the modification were classified into tumor reduction, tumor enlargement, displacement, and shape change. Results Among the 27 lesions, 55.6% required the treatment plan modification. The reasons were tumor reduction in six lesions, tumor enlargement in three lesions, displacement in three lesions, and shape change in three lesions. The planning target volume (PTV) size changed up to 43.0% and the shift of center of PTV was a maximum of 1.7 mm. The pathological status (adenocarcinoma vs others) and timing of steroid administration (prior vs after SRT start) were the predictive factors of tumor changes required the modification. Conclusions As tumor changes might occur even during short period of SRT, the treatment plan evaluation and modification were important in SRT for large brain metastases.

Purpose The detection of intestinal/rectal gas is very important during image-guided radiation therapy (IGRT) of prostate cancer patients because intestinal/rectal gas increases the inter- and intra-fractional prostate motion. We propose a deep convolutional neural network (DCNN) to detect intestinal/rectal gas in the pelvic region. Material and methods We selected 300 anterior-posterior kilo-voltage (kV) X-ray images from 30 prostate cancer patients. Thirty images were randomly chosen for a test set, and the remaining 270 images used as the training set. The intestinal/rectal gas was manually delineated on kV X-ray images and segmented. The training images were augmented by applying artificial shifts and fed into a DCNN. The network models were trained to keep the quality of the output image close to the quality of the input image by pooling and upsampling. The training set was used to adjust the parameters of the DCNN, and the test set was used to assess the performance of the model. The performance of the DCNN was evaluated using a fivefold cross-validation procedure. The dice similarity coefficient (DSC) was calculated to evaluate the detection accuracy between the manual contour and auto-segmentation. Results The DCNN was trained within approximately 17 min with a time step of 20 s/epoch. The training and validation accuracy of the models after 50 epochs were 0.94 and 0.85, respectively. The average ± standard deviation of the DSC for 30 test images was 0.85 ± 0.08. Conclusions The proposed DCNN method can automatically detect the intestinal/rectal gas in kV images with good accuracy.

Computed tomography (CT) data are required to calculate the dose distribution in a patient’s body. Generally, there are two CT number calibration methods for commercial radiotherapy treatment planning system (RTPS), namely CT number‐relative electron density calibration (CT‐RED calibration) and CT number‐mass density calibration (CT‐MD calibration). In a previous study, the tolerance levels of CT‐RED calibration were established for each tissue type. The tolerance levels were established when the relative dose error to local dose reached 2%. However, the tolerance levels of CT‐MD calibration are not established yet. We established the tolerance levels of CT‐MD calibration based on the tolerance levels of CT‐RED calibration. In order to convert mass density (MD) to relative electron density (RED), the conversion factors were determined with adult reference computational phantom data available in the International Commission on Radiological Protection publication 110 (ICRP‐110). In order to validate the practicability of the conversion factor, the relative dose error and the dose linearity were validated with multiple RTPSes and dose calculation algorithms for two groups, namely, CT‐RED calibration and CT‐MD calibration. The tolerance levels of CT‐MD calibration were determined from the tolerance levels of CT‐RED calibration with conversion factors. The converted RED from MD was compared with actual RED calculated from ICRP‐110. The conversion error was within ±0.01 for most standard organs. It was assumed that the conversion error was sufficiently small. The relative dose error difference for two groups was less than 0.3% for each tissue type. Therefore, the tolerance levels for CT‐MD calibration were determined from the tolerance levels of CT‐RED calibration with the conversion factors. The MD tolerance levels for lung, adipose/muscle, and cartilage/spongy‐bone corresponded to ±0.044, ±0.022, and ±0.045 g/cm3, respectively. The tolerance levels were useful in terms of approving the CT‐MD calibration table for clinical use.

Thermoplastic masks, used along with surgical masks, enable immobilization methods to reduce the risk of infection in patients undergoing intracranial stereotactic radiosurgery and stereotactic radiotherapy (SRS/SRT) during the COVID‐19 crisis. The purpose of this study was to investigate the feasibility of thermoplastic mask immobilization with a surgical mask using an ExacTrac system. Twelve patients each with brain metastases were immobilized using a thermoplastic mask and a surgical mask and only a thermoplastic mask. Two x‐ray images were acquired to correct (XC) and verify (XV) the patient’s position at a couch angle of 0°. Subsequently, the XC and XV images were acquired at each planned couch angle for non‐coplanar beams. When the position errors were detected after couch rotation for non‐coplanar beams, the errors were corrected at each planned couch angle until a clinically acceptable tolerance was attained. The position errors in the translational and rotational directions (vertical, lateral, longitudinal, pitch, roll, and yaw) were retrospectively investigated using data from the ExacTrac system database. A standard deviation of XC translational and rotational position errors with and without a surgical mask in the lateral (1.52 vs 2.07 mm), longitudinal (1.59 vs 1.87 mm), vertical (1.00 vs 1.73 mm), pitch (0.99 vs 0.79°), roll (1.24 vs 0.68°), and yaw (1.58 vs 0.90°) directions were observed at a couch angle of 0°. Most of patient positioning errors were less than 1.0 mm or 1.0° after the couch was rotated to the planned angle for non‐coplanar beams. The overall absolute values of the translational and rotational XV position errors with and without the surgical mask were less than 0.5 mm and 0.5°, respectively. This study showed that a thermoplastic mask with a surgical mask is a feasible immobilization technique for brain SRS/SRT patients using the ExacTrac system.

A tick infestation is one of the most common arthropod-related skin diseases in Hokkaido, the northernmost island of Japan. Ticks also act as an infectious disease vector for humans. Tick-borne encephalitis (TBE), a highly mortal central nervous system infection caused by TBE virus (TBEV), has sporadically occurred there recently. However, there have been no epidemiological data on the current surveillance of human tick bites and the prevalence of TBEV in human-feeding ticks. This study was performed to clarify those indeterminate issues. One hundred and fifty-three ixodid ticks feeding on humans were collected from 150 outpatients in Hokkaido during the season of April to August 2018. None of the cases showed any infectious symptoms. These ticks were morphologically identified to species, and a cytopathic assay on baby hamster kidney cells was carried out to detect TBEV from each tick. The tick collection consisted of 108 Ixodes persulcatus (one nymph and 107 adult females), 44 female Ixodes ovatus, and one female Haemaphysalis japonica. No tick extracts showed positive results of the cytopathic assay, suggesting the non-existence of TBEV in the present specimens. However, the survey to detect TBEV from human-feeding ticks is still important to monitor the occurrence of TBE, because human tick bites by I. ovatus, a possible vector of TBEV, are increasing even in the northern and eastern areas of Hokkaido.

Purpose To improve the hybrid VMAT treatment plan robustness for postoperative breast cancer patients considering small patient motion shifts during treatment using a 3D-CRT with a dose gradient in the junction region. Methods Locoregional supraclavicular nodes of breast cancer patients were planned using 3D-CRT and VMAT. A 3D-CRT plan with a dose gradient on the cranial side was applied by shifting the jaw to reduce hot or cold spots. The VMAT plan was optimized based on the results of the 3D-CRT plan calculation. Hybrid plans were created by the sum of the 3D-CRT and VMAT plans. To simulate patient motion, the plans were recalculated with the VMAT plan simulating isocenter superiorly (separation) or inferiorly (overlap) shifted by 1, 2, and 3 mm. The shifted plans were compared with the non-shifted plans considering the clinical target volume (CTV) (D98% or D2%). Results The D2% value of the CTV with perfectly aligned fields for the hybrid VMAT plan with high- or low-dose gradients on the 3D-CRT plan increased from 102.8%/102.9–107.2%/105.7%, 114.9%/110.9%, and 122.2%/115.5% for each 1 mm, 2 mm, and 3 mm overlapped shift, respectively. The value of D98% to the CTV with perfectly aligned fields decreased from 95.7%/95.6–90.0%/93.1%, 81.2%/88.4%, and 72.8%/83.5% for each 1 mm, 2 mm, and 3 mm separated shift, respectively. Conclusions By employing a 3D-CRT plan with a low dose gradient on the cranial side, the dose differences can be decreased. A more robust treatment delivery option can be achieved for breast cancer treatment using our proposed hybrid VMAT.

The amphibian acanthocephalan, Pseudoacanthocephalus toshimai, was considered to be an island-endemic species in Hokkaido, Japan. However, the parasite was found from Rana ornativentris, Rana tagoi, Zhangixalus arboreus, and Bufo japonicus formosus in northern Honshu (Aomori and Iwate Prefectures), which is separated from Hokkaido by the Tsugaru Strait. The mitochondrial DNA-based phylogenetic and population genetic analyses of P. toshimai showed that the northern Honshu isolates are far distantly related to the Hokkaido isolates, and that a demographic population expansion occurred in Hokkaido during the recent geological past. The rich genetic diversity of P. toshimai in northern Honshu suggests a scenario that anuran hosts invaded Hokkaido together with P. toshimai via the land bridge of the Tsugaru Strait. However, the evolutionary history of Rana pirica, a main definitive host for P. toshimai in Hokkaido, is contradictory to the introduction scenario inferred from the parasite. The finding of several geographically mismatched isolates of P. toshimai from both northern Honshu and Hokkaido suggests a possibility that the migration of the parasite infrequently occurred between the two areas even after the land bridge disappeared. More detailed information on the evolutionary history of anurans is needed to resolve the biogeographical enigma of P. toshimai.

The cryptic diversity of trematodes was evaluated in the Nagayama-Shinkawa River, an artificial canal of the Ishikari River System of Hokkaido, Japan. Numerous migratory waterfowls use the canal as a stopover point in every spring season. The lymnaeid snail, Radix auricularia, and the semisulcospirid snail, Semisulcospira libertina, colonize the static and flowing water areas, respectively. The trematode fauna of the two snails was assessed by molecular phylogenetic and population genetic analyses. Each of distinctive clades in mitochondrial DNA trees was arbitrarily set as a species. In total, 14 species of the families Diplostomidae, Echinostomatidae, Notocotylidae, Plagiorchiidae, and Strigeidae occurred in R. auricularia, wherease S. libertina harbored 10 species of the families Echinochasmidae, Heterophyidae, Notocotylidae, and Lecithodendridae and Cercaria creta, an unclassified species whose adult stage is still unknown. The species diversity of the larval trematodes could be recognized as a “hot spot”, suggesting that the seasonal visit of waterfowls is very important to spread trematodes and to keep their diversity. A high intraspecific genetic diversity was observed in the echinostomatid, notocotylid, echinochasmid, and heterophyid species, whose definitive hosts include birds. It seems likely that each of the parasite populations is always disturbed by repeated visits of waterfowls.

An unknown species of the genus Notocotylus (Digenea: Notocotylidae) was found as the larval stage from the lymnaeid snail, Radix auricularia, in a static water area of the Chubetsu River, Hokkaido, the northernmost island of Japan. A DNA barcoding identification system was applied to detect the adult stage. Through the inspection of anatid game birds in Hokkaido, Anas crecca, Anas platyrhynchos, Anas zonorhyncha, and Mareca penelope were demonstrated to serve as the definitive hosts. The detailed morphological features of the species were characterized using adults raised experimentally in immunosuppressed mice and naturally developed larvae in R. auricularia. Although the species is morphologically similar to Notocotylus attenuatus and Notocotylus magniovatus in both adult and larval stages, its taxonomic independence was confirmed by a comprehensive study based on molecular phylogeny, morphology, and ecology. Here we propose Notocotylus ikutai n. sp. for this species. The migratory behavior of the anatid hosts and the North-Eurasian distribution of R. auricularia suggest that the new species is widely distributed in the northern Far East.

The metacercarial infections of door snails (Gastropoda: Clausiliidae) with unknown species of the genus Brachylaima (Trematoda: Brachylaimidae) have recently been reported in eastern Honshu and Kyushu, Japan. A large scale snail survey was carried out to clarify their taxonomic status. From the period of 2015 to 2020, a total of 1239 land snails (768 door snails and 471 others) were collected from 32 localities in Honshu, Shikoku, and Kyushu. The resulting trematode isolates were identified as Brachylaima sp. by mitochondrial DNA barcoding. The sporocysts were found only a few from Megalophaedusa sublunellata (Clausiliidae), Tauphaedusa subaculus (Clausiliidae), and Aegista trochula (Camaenidae), while the metacercariae were frequently detected from 14 species of Clausiliidae and 2 species of other families. Although Brachylaima sp. showed a broad range of intermediate hosts, door snails seem to be very important to drive the life cycle. The gravid adults of Brachylaima sp. was experimentally raised from metacercariae using immunosuppressed mice. Morphological, phylogenetical, and ecological considerations prompted us to propose Brachylaima phaedusae n. sp. for this unknown species. The definitive hosts of the new species are completely unknown. The wide geographic distribution and high genetic diversity of the new species suggest a possibility that the definitive host is ground-foraging birds, which prefer door snails.

Through a continuous survey of trematodes in land snails of Hokkaido, the northernmost island of Japan, we have discovered four species of the genus Brachylaima (Trematode: Brachylaimidae). Among them, Brachylaima ezohelicis, Brachylaima asakawai, and Brachylaima lignieuhadrae have already been described. Each of the three species is a strict specialist in selecting a particular species of land snail as the first intermediate host. In this report, we propose the fourth species, Brachylaima succini sp. nov., based on ecological, morphological, and phylogenetic considerations. Sporocysts and metacercariae of the new species were found exclusively from Succinea lauta, which is known as an amber snail indigenous to Hokkaido. Phylogenetic trees of nuclear 28S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) demonstrated it to be distinct from the other sympatric species. Although metacercariae of the new species possessed unique morphological characters, adult worms experimentally raised from the metacercariae were similar to those of B. ezohelicis and B. lignieuhadrae. Natural definitive hosts of the new species are unknown, but the existence of common cox1 haplotypes from far-distant localities suggests a possibility that birds are involved as the definitive hosts. Findings of amber snails coinfected with both sporocysts of the new species and Leucochloridium perturbatum also support the involvement of birds.

We investigated the feasibility of a robust optimization with 6 MV X-ray (6X) and 10 MV X-ray (10X) flattening filter-free (FFF) beams in a volumetric modulated arc therapy (VMAT) plan for lung stereotactic body radiation therapy (SBRT) using a breath-holding technique. Ten lung cancer patients were selected. Four VMAT plans were generated for each patient; namely, an optimized plan based on the planning target volume (PTV) margin and a second plan based on a robust optimization of the internal target volume (ITV) with setup uncertainties, each for the 6X- and 10X-FFF beams. Both optimized plans were normalized by the percentage of the prescription dose covering 95% of the target volume (D95%) to the PTV (1050 cGy × 4 fractions). All optimized plans were evaluated using perturbed doses by specifying user-defined shifted values from the isocentre. The average perturbed D99% doses to the ITV, compared to the nominal plan, decreased by 369.1 (6X-FFF) and 301.0 cGy (10X-FFF) for the PTV-based optimized plan, and 346.0 (6X-FFF) and 271.6 cGy (10X-FFF) for the robust optimized plan, respectively. The standard deviation of the D99% dose to the ITV were 163.6 (6X-FFF) and 158.9 cGy (10X-FFF) for the PTV-based plan, and 138.9 (6X-FFF) and 128.5 cGy (10X-FFF) for the robust optimized plan, respectively. Robust optimized plans with 10X-FFF beams is a feasible method to achieve dose certainty for the ITV for lung SBRT using a breath-holding technique.