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Rabies virus (RABV) causes fatal neurological disease. Pre-exposure prophylaxis (PrEP) and post-exposure prophylaxis (PEP) using inactivated-virus vaccines are the most effective measures to prevent rabies. In Japan, HEP-Flury, the viral strain, used as a human rabies vaccine, has historically been propagated in primary fibroblast cells derived from chicken embryos. In the present study, to reduce the cost and labor of vaccine production, we sought to adapt the original HEP-Flury (HEP) to Vero cells. HEP was repeatedly passaged in Vero cells to generate ten- (HEP-10V) and thirty-passaged (HEP-30V) strains. Both HEP-10V and HEP-30V grew significantly better than HEP in Vero cells, with virulence and antigenicity similar to HEP. Comparison of the complete genomes with HEP revealed three non-synonymous mutations in HEP-10V and four additional non-synonymous mutations in HEP-30V. Comparison among 18 recombinant HEP strains constructed by reverse genetics and vesicular stomatitis viruses pseudotyped with RABV glycoproteins indicated that the substitution P(L115H) in the phosphoprotein and G(S15R) in the glycoprotein improved viral propagation in HEP-10V, while in HEP-30V, G(V164E), G(L183P), and G(A286V) in the glycoprotein enhanced entry into Vero cells. The obtained recombinant RABV strain, rHEP-PG4 strain, with these five substitutions, is a strong candidate for production of human rabies vaccine. [ABSTRACT FROM AUTHOR]

Purpose: Fertility preservation (FP) is becoming increasingly common among child, adolescent, and young-adult (CAYA) patients with cancer. However, Japan has long lacked definite estimates of utilization rates for FP services among CAYA patients with cancer, and little is known about disease/FP outcomes among users. Therefore, the Japan Society for Fertility Preservation (JSFP) launched the Japan Oncofertility Registry (JOFR) in 2018 and started the online registration of information regarding primary disease, FP, and data on prognosis and pregnancy outcomes. This study reports the analytical results of FP data registered in the JOFR as of 2021. Methods: Data about patients' primary disease(s), treatment courses, cancer and pregnancy outcomes, and specific procedures were extracted from the JOFR and analyzed. Results: In 2021, 1244 patients received counseling or treatment related to FP (540 males, 704 females). While the numbers of males in each age group were approximately equal, most females were aged between 31 and 40 years. In total, 490 male and 540 female patients underwent FP procedures. Leukemia, testicular cancer, and malignant lymphoma accounted for the majority of male cases seeking treatment, whereas breast cancer was the primary disease in two-thirds of the females. Since 1999, 395 patients have accumulatively experienced subsequent pregnancy. Conclusions: As of January 2022, >7000 cases from >100 fertility facilities have been registered in the JOFR. In the future, maintaining JOFR to disseminate information on cancer prognoses, pregnancy rates, and other oncofertility outcomes is expected to drive further expansion of oncofertility services in Japan. [ABSTRACT FROM AUTHOR]

Background: Multiple surveys of the general public and experts on human genome editing have been conducted. However, many focused only on editing in clinical applications, with few regarding its use for basic research. Given that genome editing for research purposes is indispensable for the realization of clinical genome editing, understanding lay attitudes toward genome editing in research, particularly using human embryos, which is likely to provoke ethical concerns, is helpful for future societal discussion. Methods: An online survey was conducted with Japanese laypeople and researchers to ascertain their views regarding human genome editing for research purposes. Participants were queried about their acceptance as a function of the target of genome editing (germ cells, surplus IVF embryos, research embryos, somatic cells); then, those who answered "acceptable depending on the purpose" were asked about their acceptance in the context of specific research purposes of genome editing. Participants were also asked about their expectations and concerns regarding human genome editing. Results: Replies were obtained from 4,424 laypeople and 98 researchers. Approximately 28.2-36.9% of the laypeople exhibited strong resistance to genome editing for research purposes regardless of their applications. In contrast, 25.5% of the researchers demonstrated resistance only to genome editing in research embryos; this percentage was substantially higher than those concerning the other three targets (5.1-9.2%). Approximately 50.4-63.4% of laypeople who answered "acceptable depending on the purpose" approved germline genome editing for disease research; however, only 39.3-42.8% approved genome editing in basic research to obtain biological knowledge. In contrast, the researchers displayed a lower degree of acceptance of germline genome editing for research purposes related to chronic diseases (60.9-66.7%) than for other research purposes (73.6-90.8%). Analysis of responses concerning expectations and concerns indicated that laypeople who would not accept genome editing of human embryos did not necessarily worry about "instrumentalization of the embryo." They also had substantially low expectations for recognized advantages of genome editing, including "advances in science" and "reduction of intractable diseases," compared with other groups of respondents. Conclusion: The assumptions shared among experts in conventional bioethical debates and policy discussions on human genome editing are not self-evident to laypeople. [ABSTRACT FROM AUTHOR]

Apostichopus japonicus is preserved in dried form after processing and used as food and traditional medicine. During the sea cucumber drying process, the boiled liquid is discarded as waste. In this study, the freeze-dried powder obtained from the boiled aqueous extracts from sea cucumbers cultured in Japan and Taiwan (SCAE-Tw and SCAE-Jp) were used to observe the anti-melanogenesis effects of the extracts on B16F10 cells and zebrafish embryos. A foam test, Fourier transform infrared (FTIR), and ultraviolet (UV) spectra analysis revealed that SCAEs had amphiphilic features similar to saponin. According to the saponin standard, we calculated that SCAE-Tw and SCAE-Jp contained 3.45 and 6.48 mg, respectively, of the saponin component per 100 mg after the lyophilization treatment. The survival rate experiments indicated that the lethal concentration in B16F10 cells is 54 and 201 μg/mL, respectively. The anti-melanogenic activity of the SCAEs was confirmed by assessing the inhibition of melanin content and intracellular tyrosinase activity in B16F10 cells and zebrafish embryos. qPCR analysis demonstrated that SCAEs suppressed the expression of Mitf, Tyr, and Trp-1 in a concentration-dependent manner. This research may provide new insights into the reuse of food processing wastewater and the development of novel anti-melanin treatments and applications in aquaculture, cosmetics, and medicine. [ABSTRACT FROM AUTHOR]

Purpose: The Japan Society of Obstetrics and Gynecology conducted a nationwide clinical study to evaluate the pregnancy outcomes of preimplantation genetic testing for aneuploidy or chromosomal structural rearrangement (PGT‐A/SR). Methods: Patients that had experienced recurrent implantation failure, recurrent pregnancy loss, or chromosomal structural rearrangement were recruited from 200 fertility centers in Japan. For patients in whom one or more blastocysts were classified as euploid or euploid with suspected mosaicism, a frozen–thawed single embryo transfer (ET) was performed. Results: A total of 10 602 cycles, maternal age 28–50 years, were enrolled in this study. 42 529 blastocysts were biopsied, and 25.5%, 11.7%, and 61.7% of embryos exhibited euploidy, mosaicism, and aneuploidy, respectively. At least one euploid blastocyst was obtained in 38.3% of egg retrieval cycles with embryo biopsy. A total of 6080 ETs were carried out, and the clinical pregnancy rate per ET, ongoing pregnancy rate per ET, and miscarriage rate per pregnancy were 68.8%, 56.3%, and 10.4%, respectively. The rates of clinical pregnancy and miscarriage remained relatively constant across all maternal ages. Conclusions: Preimplantation genetic testing for aneuploidy or chromosomal structural rearrangement may improve the pregnancy rate per ET and reduce the miscarriage rate per pregnancy, especially in patients of advanced maternal age. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study is to compare anthropometric measurements between term singletons conceived via fresh embryo transfer (FreET) and frozen embryo transfer (FET) and those born via natural conception (NC) or fertility treatments milder than assisted reproductive technology (non‐ART) at 6 years of age. Methods: A total of 8149 children were enrolled, and questionnaires about anthropometric measures (weight, height, BMI) were addressed to parents, when the children were 1.5, 3, and 6 years of age. A total of 3299 term singletons were enrolled at birth: 533, 476, 916, and 1374 in the NC, non‐ART, FreET, and FET groups, respectively. Results: A total of 1635 term singletons (290, 176, 467, and 702 in the NC, non‐ART, FreET, and FET groups respectively) were enrolled until 6 years of age (follow‐up rate, approximately 50%). When non‐ART group was used as control, the FreET children were 1.0 cm taller than the non‐ART children at 6 years of age, after adjusting for confounding factors. However, no differences were observed in the anthropometric data among the non‐ART, ART, and NC children at 6 years of age. Conclusion: At 6 years of age, term singletons were taller in the FreET group than in the non‐ART group, after adjusting for confounders. [ABSTRACT FROM AUTHOR]

The rabies virus is widely distributed and vaccines are an important strategy to prevent its spread. The whole-genome sequences of rabies strains in relation to vaccine development provide essential information to maintain vaccine quality and develop new vaccines. However, the genetic characteristics of the purified chick embryo cell culture rabies vaccine, KM Biologics (PCECV-KMB), developed in Japan in the 1970s, have not been explored. In this study, we conducted a genome-wide analysis of the open reading frame regions of rabies strains discovered from the 1940s–1980s and used to develop chick embryo cell-adapted HEP-Flury small plaque-forming (CEF-S) strain, which is a vaccine strain of PCECV-KMB. The genetic characteristic of CEF-S, developed by acclimation of the HEP-Flury-NIID strain to one-day eggs and subsequently to chick embryo cells, were confirmed by comparing the genome identity and revealing the nine amino acid mutations between CEF-S and HEP-Flury-NIID. The efficacy of PCECV-KMB was evaluated using attack strains isolated in Thailand in the 1960s–1970s during vaccine development. Phylogenetic analyses of the attack strains classified them in the same Asian clade as the 2000s imported cases from the Philippines to Japan, suggesting that PCECV-KMB is adequate for preventing the spread of the current rabies virus. [ABSTRACT FROM AUTHOR]

Ovarian stimulation is a key issue in assisted reproductive technology (ART) treatment. ART practice in Japan is unique with various types of ovarian stimulation protocols, which may contribute to lower pregnancy rates compared to other countries. This review aims to clarify optimal individualized ovarian stimulation for improving the pregnancy rate per one oocyte retrieval cycle in Japan. We performed a literature review to describe ovarian stimulation, classification of infertile women depending on ovarian reserve and response, and Japanese ART data and discussed optimal conventional and mild ovarian stimulation protocols in Japan. According to Japanese ART registry data, the live birth rate of 30–35‐year‐old women was 32%–37% per ET cycle; therefore, four to five embryos are calculatedly needed when aiming a cumulative live birth rate of ≥80%. Mild stimulation aimed at collecting 5–10 oocytes can be alternative choice as an optimal ovarian stimulation protocol in young women. In 40‐year‐old women, the live birth rate is 18.8%, resulting in eight or more embryos as necessary. Conventional stimulation must be required in women with advanced age. In poor responders, however, mild stimulation may be sufficient for maximumly extracting their ovarian function. In Japan, mild ovarian stimulation can be selected in patients with a good prognosis and poor responders; however, conventional ovarian stimulation is necessary for women in advanced age. [ABSTRACT FROM AUTHOR]

The Pacific oyster (Crassostrea gigas) is an important species in oyster farming worldwide, including in Japan. Hiroshima Bay is one of the most important oyster farming areas in Japan. We investigated the occurrence of antifouling biocides used worldwide including diuron, Irgarol 1051 (Irgarol), and 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT), which have been detected at sub-ppb levels in seawater in Japan, and estimated their no observed effect concentrations (NOECs). In recent years, the spat settlement of Pacific oysters has become poor, which presents a challenge for oyster aquaculture in Hiroshima Bay; hence, we conducted embryotoxicity and larva settlement tests using Pacific oysters. Compared to diuron and Irgarol, DCOIT exhibited a higher toxicity toward oyster embryos, and the minimum 24-h NOEC toxicity value for the oyster embryos was <3 ng/L. The highest concentrations of diuron, Irgarol, and DCOIT in the environmental seawater in the Seto Inland Sea were 27.6, 3.2, and 24 ng/L, respectively. Considering the NOECs, the environmental concentrations of these biocides suggest that the ecological risks posed by diuron and Irgarol are low, whereas those posed by DCOIT are high. However, the rate of detection of DCOIT was low because it degraded rapidly in the seawater before treatment for chemical analysis, except in the case of the treatment on the research vessel. [ABSTRACT FROM AUTHOR]

Seasonal occurrences of marine cladocerans in offshore Suruga Bay, Japan, were studied from 2014 to 2019. Zooplankton samples were collected monthly from a station (depth: ca. 1000 m) located in the offshore area of the bay. Cladocerans appeared each year from February to December, and disappeared in January. Their abundance increased drastically from April to September and comprised a substantial portion of the offshore mesozooplankton community during this period. Maximum cladoceran abundance occurred from June to August, ranging from 65.9-1341.9 individuals m-3. These patterns in cladoceran abundances were basically repeated each year throughout the study period. This result suggests that mass occurrences of cladocerans in offshore regions of the bay during the spring-summer are regular events rather than sporadic. Seven species, which have previously been reported from Japanese waters, were identified, and successional changes in the dominant species were observed. The most abundant species, Penilia avirostris, carried parthenogenetic embryos in brood pouches, suggesting that they were not simply transported from coastal areas, but that they also reproduce in the offshore waters. Regular mass occurrences of marine cladocerans in offshore Suruga Bay could have an important impact on the offshore ecosystem of the bay, and factors enabling these population cycles need to be determined. [ABSTRACT FROM AUTHOR]

Toads of the Family Bufonidae possess neurotoxins (bufadienolides) which are generally considered to be synthesized de novo. Thus, invasive toad species pose a threat to native predators via toxic effects. However, the influence of diet on toad toxicity is poorly understood. We evaluated the effect of diet on toxicity of embryos of the invasive toad Bufo japonicus formosus in Japan, using native tadpoles (Rana pirica) as a bioassay. Specifically, we compared the toxicity of embryos spawned by captive toads reared entirely on non-toxic prey versus embryos spawned by wild toads. Embryos of captive-reared toads and wild toads were comparable in terms of toxic effects: all tadpoles that consumed a toad embryo died, irrespective of origin of embryos. Embryo toxicity appears to be via parental (likely maternal) provisioning. Our study indicates that adult B. j. formosus produce toxins (likely bufadienolides) in amounts sufficient for embryos to be fatal to native predator tadpoles irrespective of whether the adult diet contains toxins. Thus, the toxic effects of invasive B. j. formosus on R. pirica, and possibly on other sensitive native predator species, are likely to be widespread and occur regardless of variation in prey biota in invaded areas in Japan. [ABSTRACT FROM AUTHOR]

Copyright of Avian Diseases is the property of American Association of Avian Pathologists, Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objectives: Haemaphysalis longicornis is the most important tick species in Japan and has a wide range of vector capacity. Due to its veterinary and medical importance, this tick species has been used as a model for tick/vector biological studies. To identify the key molecules associated with physiological processes during blood feeding and embryogenesis, full-length cDNA libraries were constructed using the fat body, hemocytes-containing hemolymph, midgut, ovary and salivary glands of fed females and embryos of the laboratory colony of parthenogenetic H. longicornis. The sequences of cDNA from the salivary glands had been already released. However, the related information is still poor, and the other expressed sequence tags have not yet been deposited. Data description: A total of 39,113 expressed sequence tags were obtained and deposited at the DNA DataBank of Japan. There were 7745 sequences from embryos, 7385 from the fat body, 8303 from the hemolymph including hemocytes, 7385 from the midgut, and 8295 from the ovary. The data, including expressed sequence tags from the salivary glands was summarized into Microsoft Excel files. Sharing this data resource with the tick research community will be valuable for the identification of novel genes and advance the progress of tick research. [ABSTRACT FROM AUTHOR]

The large Japanese field mouse (Apodemus speciosus) is a small rodent species endemic to Japan. The genetic characteristics of A. speciosus include different chromosome numbers within the same species. Furthermore, A. speciosus has been used in radiation and genetic research. In the present study, a pregnant A. speciosus was obtained, and histochemical analysis of the implanted embryos was performed and compared with the developmental stages of the mouse (Mus musculus). Although there were some differences, the structures of the implanted embryos, including the primitive streak and placenta of A. speciosus were similar to those of mouse. Our study will be important for the construction of a developmental atlas of A. speciosus. [ABSTRACT FROM AUTHOR]

Aim: To investigate the clinical factors and factors that affect the decisions regarding storage of cryopreserved embryos obtained using assisted reproductive technology. Methods: Clinical characteristics affecting the decisions regarding cryopreserved embryos were analyzed in 5724 Japanese couples who underwent in vitro fertilization (IVF) or intra‐cytoplasmic sperm insemination (ICSI) and embryo transfer over 4 years since April 2015 at our clinic. Statistical analysis was carried out using JMP software. Results: The number of oocytes retrievals and embryos stored, outcomes and number of children, and age of the female patients and male partners were related to the decision‐making regarding cryopreserved embryos. Childbearing and no wish for another child were the major reasons for discontinuing embryo storage. The number of oocytes retrievals and embryos in storage, age of the female patients, and sex of the child were independently associated with this decision‐making in 2682 patients with a single child. Women with male children were more likely to choose discontinuation of embryo storage than those with female children. Conclusion: Already having a child and not wishing for further treatment due to age along with the presence of a male child affect the decision to continue or discontinue embryo storage in Japanese patients with infertility. [ABSTRACT FROM AUTHOR]

The two major components of the metanephros, the urinary collecting system (UCS) and nephron, have different developmental courses. Nephron numbers vary widely between species and individuals and are determined during fetal development. Furthermore, the development of nascent nephrons may contribute to the expansion of the proximal part of the UCS. This study investigated the distribution of nascent nephrons and their interrelationship with UCS branches during human embryogenesis. We obtained samples from 31 human embryos between Carnegie stages (CSs) 19 and 23 from the Kyoto Collection at the Congenital Anomaly Research Center of Kyoto University in Japan. Serial histological sections of the metanephros with the UCS were digitalized and computationally reconstructed for morphological and quantitative analyses. The three‐dimensional (3D) coordinates for the positions of all UCS branch points, end points, attachment points to nascent nephrons (APs), and renal corpuscles (RCs) were recorded and related to the developmental phase. Phases were categorized from phase 1 to phase 5 according to the histological analysis. The UCS branching continued until RCs first appeared (at CS19). End branches with attached nascent nephrons (EB‐AP[+]) were observed after CS19 during the fifth generation or higher during the embryonic period. The range of end branch and EB‐AP(+) generation numbers was broad, and the number of RCs increased with the embryonic stage, reaching 273.8 ± 104.2 at CS23. The number of RCs connected to the UCS exceeded the number not connected to the UCS by CS23. The 3D reconstructions revealed RCs to be distributed around end branches, close to the surface of the metanephros. The RCs connected to the UCS were located away from the surface. The APs remained near the end point, whereas connecting ducts that become renal tubules were found to elongate with maturation of the RCs. Nascent nephrons in RC phases 3‐5 were preferentially attached to the end branches at CS22 and CS23. The mean generation number of EB‐AP(−) was higher than that of EB‐AP(+) in 19 of 22 metanephros and was statistically significant for eight metanephros at CS22 and CS23. The ratio of the deviated branching pattern was almost constant (29%). The ratio of the even branching pattern with EB‐AP(+) and EB‐AP(+) to the total even branching pattern increased with CS (9.2% at CS21, 19.2% at CS22, and 45.4% at CS23). Our data suggest the following: EB‐AP(+) may not branch further at the tip (i.e., by tip splitting), but branching beneath the AP (lateral branching) continues throughout the embryonic stages. Our study provides valuable data that can further the understanding of the interactions between the UCS and nascent nephrons during human embryogenesis. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the risk of ectopic pregnancies (EPs) for fresh cycles according to different ovarian stimulation protocols.Design: Registry-based retrospective cohort study.Setting: Not applicable.Patient(s): A total of 68,851 clinical pregnancies after fresh single embryo transfer between 2007 and 2015.Intervention (s): None MAIN OUTCOME MEASURE(S): Ectopic pregnancies. Odds ratios and 95% confidence intervals for EPs were calculated by using generalized estimating equations adjusted for potential maternal and treatment characteristics.Result(s): Among 68,851 clinical pregnancies, 1,049 (1.46%) cases of EP were reported. Compared with natural cycles, all ovarian stimulation protocols were associated with a significantly increased risk of EP. Ovarian stimulation with clomiphene (CC) demonstrated the highest odds ratios for EPs. Significant associations between ovarian stimulation protocols and EP compared with natural cycles were prominent when the number of retrieved oocytes was low (1-3) to moderate (4-7), but there were no significant associations when the number of retrieved oocytes was high (≥8).Conclusion(s): Ovarian stimulation protocols were significantly associated with an increased risk of EP. In particular, ovarian stimulation with CC had the highest risk of EP compared with other stimulation protocols. Further studies are essential to investigate possible confounding factors for different ovarian stimulation protocols, especially CC, and the risk of EP. [ABSTRACT FROM AUTHOR]

Copyright of Magyar Állatorvosok Lapja is the property of Herman Otto Intezet Nonprofit Kft. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this article, I discuss the significance of embryological knowledge, such as the red and white drops and the five developmental stages of the embryo, in medieval Shingon esoteric Buddhism. Specifically, I examine the writings of Kakuban, an eminent Shingon Buddhist monk in early medieval Japan, and point out that, according to Kakuban, embryological knowledge was connected with the six elements, which were fundamental to Shingon conceptions of ontology. In other words, by constructing embryological theories, medieval Shingon monks such as Kakuban attempted to make a correlation between abstract and distant cosmologies and the life and death realities of their daily lives. [ABSTRACT FROM AUTHOR]

Aim: The survival rates of cancer patients have greatly improved owing to the advances in oncology. The preservation of fertility in cancer patients is an important task. To determine the reality of cryopreservation of embryos, oocytes and ovarian tissue in cancer patients, large‐scale survey analysis was performed in Japan. Methods: We sent 613 Japan Society of Obstetrics and Gynecology‐certified assisted reproductive technology institutions a questionnaire about their experience of performing cryopreservation for cancer patients between January 2011 and December 2015. Subsequently, the institutions that conducted cryopreservation for cancer patients were sent a second questionnaire. Results: We received replies from 481 (78.5%) institutions. Among them, 126 (26.2%) conducted cryopreservation for cancer patients. These 126 institutions were sent a second questionnaire. Of these, 108 (85.7%) institutions responded. At the 108 institutions, 1085 embryo or oocyte cryopreservation procedures and 122 ovarian tissue cryopreservation procedures were conducted for cancer patients. Cryopreservation was mainly performed for breast cancer patients (~70%), followed by patients with hematological malignancy. A total of 361 and 19 embryo transfer cycles were performed for patients whose embryos and oocytes were cryopreserved, respectively, and 42 and seven institutions reported pregnancy outcomes after embryo transfer in patients that underwent embryo and oocyte cryopreservation, respectively. However, pregnancy was not observed in the seven cases that underwent ovarian tissue transfer. Conclusion: Indications, age limits and ovarian stimulation protocols for cryopreservation widely varied between the institutions. A national registration system for oncofertility must be established to evaluate the safety and efficacy of the current system. [ABSTRACT FROM AUTHOR]

Purpose: In assisted reproductive technology, normal zygotes are bipronuclear (2PN) during fertilization confirmation; however, sometimes, nonpronuclear zygotes (0PN) and monopronuclear zygotes (1PN) are found during routine observations. Methods: To elucidate the clinical usefulness of in vitro‐fertilized embryos, we investigated the rates of clinical pregnancy, live birth, miscarriage, and congenital abnormality after transfer of frozen‐thawed 1PN‐ and 0PN‐derived single blastocysts at Denentoshi Ladies Clinic, Kanagawa, Japan. Results: The rates of pregnancy and live birth for 1PN‐derived blastocysts obtained by conventional in vitro fertilization were 37.5% and 27.1%, respectively, which was not significantly different from those for 2PN‐derived blastocysts; however, the rates for 0PN‐derived blastocysts were significantly lower. The pregnancy and live birth rates for 0PN‐derived embryos obtained by intracytoplasmic sperm injection (ICSI) were 45.7% and 34.8%, respectively, which was not significantly different from those for 2PN‐derived blastocysts; however, the rates for 1PN‐derived blastocysts were significantly lower (4.0% for both) than those for 2PN‐ and 0PN‐derived blastocysts. No congenital abnormalities were found in infants resulting from transfer of 0PN‐ or 1PN‐derived blastocysts. Conclusions: Both 1PN‐ and 0PN‐derived blastocysts can be used for embryo transfer; however, care should be taken in making decisions about 1PN‐derived blastocysts, especially if they are obtained by ICSI. [ABSTRACT FROM AUTHOR]

Purpose: To demonstrate the benefits of the freeze‐all strategy for in vitro fertilization treatment based on retrospective analyses. Methods: Post‐thaw embryo survival rates of slow‐frozen embryos in 294 cycles and vitrified embryos in 12 195 cycles were assessed. Progesterone (P4) and estradiol (E2) levels per mature oocyte by age category were assessed in 9081 cycles and pregnancy rates with fresh embryo transfer and frozen‐thawed embryo transfer by P4 level were assessed in 1535 cycles. Results: The survival rates of frozen‐thawed embryos were 92.5% with slow freezing and 99.1% with vitrification. P4 levels on the day of human chorionic gonadotropin (hCG) injection showed a trend toward an increase with age. The pregnancy rate per mature oocyte with fresh embryo transfer decreased dependently upon P4 level, while that with frozen‐thawed embryo transfer was not affected by P4 level. The pregnancy rates with frozen‐thawed embryo transfer were higher than those with fresh embryo transfer in patients aged 42 years or younger. Conclusions: The freeze‐all strategy is a valuable treatment option which allows the separation of an embryo transfer cycle from an oocyte retrieval cycle, especially for patients with high P4 levels at oocyte retrieval and patients of advanced maternal age. [ABSTRACT FROM AUTHOR]

The use of human embryos in genome editing research has recently been approved in China and the United Kingdom. In Japan, the debate on regulations on genome editing research studies using human embryos is underway, but is becoming increasingly entangled, to the point of deadlock. One main reason for this is the misalignment between the Japanese government and the research communities, in their awareness surrounding these regulations. In this article, we report on this ongoing and entangled debate in Japan concerning the regulations on genome editing technology using human embryos. The most critically needed next step is a grassroots level discussion among various experts such as those in the arts and humanities. [ABSTRACT FROM AUTHOR]

How are tiger shark embryos nourished to large size without a placental connection? Tiger sharks belong to the family Carcharhinidae, and all carcharhinid sharks are placental with the exception of the tiger shark. The aim of this study was to test the hypothesis that tiger shark embryos are nourished to large size by imbibing a clear uterine fluid found in their egg cases. Based on weights of fertilized eggs and of term embryos, the tiger shark is a matrotrophic species, and its embryos appear to reach gains of 2119% in wet weight and 1092% in dry weight during gestation. By measuring the total energy content of the fluid in the egg case by chemical oxygen demand (COD), the authors demonstrate that clear liquid in the tiger shark egg case is an energy-rich embryotrophe that nourishes the embryos to large size. We suggest that the process be termed ‘embryotrophy'. The process appears to be an adaptation for producing large broods of large embryos in a species lacking a placental connection. [ABSTRACT FROM PUBLISHER]

The life history, reproductive ecology and habitat utilization of the Itasenpara (deepbody) bitterling Acheilognathus longipinnis were investigated in a lowland segment of the Moo River in Toyama Prefecture, central Honshu, Japan. Analysis of 1285 individuals revealed that the study population comprised a single size class, an age at maturation of 3 months and a life span of 1 year. On the basis of the growth pattern, the life cycle was divided into two stages: the juvenile stage, characterized by rapid growth, and the adult stage at which growth ceased. Spawning by A. longipinnis was recorded between early September and late October. Female A. longipinnis in the 0+ year age class began to mature when they reached a standard length ( LS) of 56·4 mm. Mature females had a large clutch size (maximum 273 eggs) and deposited highly adhesive and relatively large eggs (2·55 mm3; major axis, 3·12 mm; minor axis, 1·22 mm) via a short ovipositor (mean length, 21·5 mm) into freshwater mussels. The embryos remained in the gill cavities of the freshwater mussels (used as a spawning substratum) and emerged as juveniles ( LS, 9 mm). Habitat utilization during spawning was analysed using a generalized linear model. The best-fit model showed that three environmental factors (freshwater mussel availability, water depth and vegetation cover) were important variables for habitat utilization by A. longipinnis. Shallow areas (water depth, 250-330 mm) created for rice paddy management and areas with an abundance of cover were particularly effective for predator avoidance. These results suggest that maintenance of water level fluctuations corresponding with rice cultivation and the abundance of vegetation on the river bank (particularly avoidance of concrete revetments) is essential for conservation of this species under current practices for rice cultivation in Japan. [ABSTRACT FROM AUTHOR]

The dose response of embryos of the zebrafish, Danio rerio , irradiated at 5 h post fertilization (hpf) by 2-MeV neutrons with ≤100 mGy was determined. The neutron irradiations were made at the Neutron exposure Accelerator System for Biological Effect Experiments (NASBEE) facility in the National Institute of Radiological Sciences (NIRS), Chiba, Japan. A total of 10 neutron doses ranging from 0.6 to 100 mGy were employed (with a gamma-ray contribution of 14% to the total dose), and the biological effects were studied through quantification of apoptosis at 25 hpf. The responses for neutron doses of 10, 20, 25, and 50 mGy approximately fitted on a straight line, while those for neutron doses of 0.6, 1 and 2.5 mGy exhibited neutron hormetic effects. As such, hormetic responses were generically developed by different kinds of ionizing radiations with different linear energy transfer (LET) values. The responses for neutron doses of 70 and 100 mGy were significantly below the lower 95% confidence band of the best-fit line, which strongly suggested the presence of gamma-ray hormesis. [ABSTRACT FROM AUTHOR]

In 2002, a commercial layer flock in Japan was initially diagnosed as being infected with infectious bronchitis (IB) based on clinical signs, virus isolation, and serological analysis but was later found to be atypically infected with velogenic Newcastle disease virus (NDV) following molecular diagnosis. The flock had slightly decreased egg production and an increased occurrence of soft-shelled eggs without significant mortality. IB-like viruses were isolated, which caused dwarfing and curling in 12-day-old chicken embryos. Ten years after this case, retrospective genetic analyses showed that apart from IB virus (IBV), the flock was also infected with NDV. Mean death time (MDT), intracerebral pathogenicity index (ICPI), and deduced amino acid sequence of the cleavage site of the fusion (F)-protein gene revealed that the NDV isolate was velogenic (112RRQKR116). These results indicate that poultry clinicians should look out for atypical velogenic ND, especially in vaccinated commercial chicken flocks, which may harbor hidden NDV infection. [ABSTRACT FROM AUTHOR]

The role of women is essential in embryo creation and donation, as they undergoin vitrofertilisation (IVF) treatment for ova collection. Yet, it has often been pointed out that in the process of embryo donation women's role is largely neglected, as if embryos come from nowhere. The procedure of embryo donation in Japan is a case in point. During field research in Japan, we found that women were of the view that the procedures of embryo donation are inhuman, cold and harsh, while specialists said that no value is attached to embryos in Japan and that women have no scruples about donating them. This article explores the cause of the gap in the perception of experiences of ova collection between women and IVF specialists. Our primary source of data was interview narratives with women undergoing IVF, IVF specialists, nurses, counsellors and policy makers. Field research was conducted in 2006–2008. We analysed the process and the ways in which women's experiences of ova collection become invisible in Japanese socio-cultural contexts, including marriage, households and motherhood values in Japan. Our analysis critically draws on Marxist feminist theories, which enabled us to discuss the link among the production of embryos and ova, reproduction, ownership and gender. This theoretical framework also enabled us to view the case of Japan, paying particular attention to historical perspectives and specific cultural forms. [ABSTRACT FROM PUBLISHER]

Prohibition of Ot (organotin) compounds was introduced in Japan in 1997 and worldwide from September 2008. This meant that the production of paints containing TBT compounds was stopped and alternatives to the available Ot antifoulants had to be developed. It has been claimed that the degradation by-products of these alternative antifoulants were less toxic than those of Ot compounds. Since the introduction of the alternative antifoulants, the accumulation of these compounds has been reported in many countries. However, the toxicity of these compounds was still largely unreported. In this research, the toxicity of the alternative Ot antifoulants TPBP (triphenylborane pyridine) and TPBOA (triphenylborane octadecylamine) and their degradation products on Crassostea gigas and Hemicentrotus pulcherrimus were tested. The results showed that toxic effects in Crassostea gigas was higher for each antifouling biocide than that in Hemicentrotus pulcherrimus. Also, while the toxicity of the Organoboron antifoulants and the Ots were the same, the former's degradation products were much less harmful. [ABSTRACT FROM AUTHOR]

Abstract: To carry out a primary risk assessment in the Inland Sea of Japan for 2,2′-dipyridyldisulfide [(PS)2], a metal pyrithione photodegradation product, we used a methodology based on the species sensitivity distribution (SSD) estimated with a Bayesian statistical model. We first conducted growth inhibition tests with three marine phytoplankton species, Tetraselmis tetrathele, Chaetoceros calcitrans, and Dunaliella tertiolecta. We also performed acute and early life stage toxicity (ELS) tests with a teleost fish, the mummichog (Fundulus heteroclitus). The algal growth inhibition tests revealed that the 72-h EC50 ranged from 62 to 1100μg/L. Acute toxicity tests with larval mummichogs revealed that the 96-h LC50 was approximately 500μg/L based on the actual toxicant concentrations. ELS testing of (PS)2 under continuous flow-through conditions for 50 days revealed that growth was the most sensitive endpoint, and both total length and body weight were significantly lower in the groups exposed to 27μg/L (PS)2 compared to the solvent control group. We determined a lowest observed effect concentration of 17μg/L and a NOEC of 5.9μg/L based on the actual toxicant concentrations. By using the ecotoxicity data (LC50 and EC50) from this study and previous work, we calculated a hazardous concentration that should protect 95% and 99% of species (HC5 and HC1) based on the SSD derived with a Bayesian statistical model. The medians with 90% confidence intervals (parentheses) of the HC5 and HC1 were 31.0 (3.2, 101.8)μg/L and 10.1 (0.5, 44.2)μg/L, respectively. In the ELS test, about 80% of hatched larvae exposed to 243-μg/L (PS)2 displayed a notochord undulation. To elucidate the cause of the notochord undulation, we carried out embryo toxicity tests by exposing embryos at various developmental stages to (PS)2. Exposure to (PS)2 through the entire gastrulae stage was important to induction of the morphological abnormality. Lysyl oxidase activity was significantly decreased in these embryos compared to the control group, a suggestion that lysyl oxidase-mediated collagen fiber organization, which is essential for notochord formation, is disrupted because of (PS)2 toxicity. We also investigated the occurrence of (PS)2 in water from several coastal sites of the Inland Sea and detected (PS)2 at concentrations of <0.1–0.4ng/L. Comparison of environmental concentrations to the HC values suggests that the current ecological risk posed by (PS)2 in the Inland Sea is low. This is the first report of the detection of a metal pyrithione degradation product in the natural marine environment. [Copyright &y& Elsevier]

To obtain data concerning the survival of embryos and calves derived from somatic cell nuclear transfer (SCNT) in Japan, a nationwide survey was carried out in April, 2009. As a result, data concerning 3264 embryo transfers (ETs) with SCNT embryos which produced 301 calves were accumulated and their survival was analyzed. The present survey revealed that survival rates of transferred bovine embryos and produced calves derived from SCNT had not improved over a decade (1998-2007). A remarkable feature of the pregnancies with SCNT embryos was a high incidence of spontaneous abortions. When the decade was divided by the occurrence of bovine spongiform encephalopathy (BSE) in 2001, significant decreases in the 'after BSE' period (2002-2007) were observed in the percentages of calves born ( P < 0.01), calves living at birth ( P < 0.05), calves living for 24 h ( P < 0.05) and 6 months ( P < 0.01). Abortions that occurred during 61-99 days after ETs were significantly increased ( P < 0.01) in the 'after BSE' period. Certain kinds of regeneration that occurred in oocytes during the 15-20 h of storage of bovine ovaries at 10-15°C as a part of BSE inspection might have had some negative effects on SCNT embryos when these oocytes were used as recipients of SCNT. [ABSTRACT FROM AUTHOR]

Abstract: This study investigated the sub-lethal effects of waterborne mercury on growth, bioaccumulation and antioxidative responses of larvae and juveniles of Japanese flounder (Paralichthys olivaceus). Fish were exposed to 0–10μg Hg2 + L−1 solutions from embryonic to the juvenile stages for 80days. Antioxidative responses to mercury exposure were studied in metamorphosing larvae (18days post hatching, dph), settling larvae (33dph) and juveniles (78dph). Results showed that increasing mercury concentration led to increased mercury bioaccumulation and reduced flounder growth. Of the antioxidants investigated, superoxide dismutase (SOD) and catalase (CAT) activities at the three developmental stages were sensitive to mercury exposure and increased with increasing mercury concentration. Glutathione (GSH) content was elevated in metamorphosing larvae, but decreased in juveniles as mercury concentration increased. Glutathione-S-transferase (GST) activity did not significantly vary with mercury concentration in either larvae or juveniles. Mercury exposure did not affect malondialdehyde (MDA) content of larvae, but significantly increased MDA content of juveniles. Results suggest that flounder larvae and juveniles have the potential to manipulate the levels of antioxidants such as SOD, CAT and GSH, which protect flounder from oxidative stress induced by mercury exposure. These antioxidants could serve as biomarkers of mercury contamination in the aquatic environment. [Copyright &y& Elsevier]

Spatiotemporal recruitment patterns of scleractinian corals were investigated around Iriomote Island, Ryukyu Archipelago, Japan, in relation to adult coral cover in 2005 and 2006. Although almost all corals were broadcasting spawners, the relationship between recruitment and adult coral cover differed among coral families (Acroporidae, Poritidae, and Pocilloporidae), likely due to differences in embryonic development time. For spawning pocilloporid corals, whose larvae develop relatively more rapidly, recruitment was higher at sites where adult coral cover was higher. In contrast, recruitment was not related to adult coral cover in acroporid and poritid corals, whose embryonic development times were relatively slow. Moreover, recruitment of acroporid corals varied between years, and recruitment was greater at leeward compared to windward reefs for a few days after spawning. These results suggest that embryonic development time and wind-driven surface currents affect larval dispersal and subsequent recruitment patterns at a local scale. Based on embryonic development time, some spawning corals are more likely to have higher rates of self-seeding than others. Our results predict that among spawning corals, local populations of acroporid and poritid corals, whose larvae potentially disperse over long distances and recruit in neighboring reefs, are more resilient to local disturbances than those of pocilloporid corals, whose recruitment relies upon local stock. [ABSTRACT FROM AUTHOR]

We examined the efficacy and safety of the Japanese purified chick embryo cell rabies vaccine (PCECK) when administered on days 0, 7, and 28, as recommended by the WHO. Post-vaccination serum samples were obtained from 53 human subjects, and rabies antibody titers were determined by a combination of enzyme-linked immunosorbent assay (ELISA) and neutralizing antibody (NA) assay. By day 42 of the experiment, which was 2 weeks after the third dose, all subjects had developed NA titers of 0.5 IU/ml or higher. The geometric mean titers of ELISA antibody and NA were 3.8 EU/ml and 5.7 IU/ml, respectively. Overall, the vaccine was well tolerated by all subjects. These results suggest that PCEC-K used for pre-exposure immunization according to the WHO schedule is as immunogenic and effective as the current pre-exposure immunization regimen in Japan, which consists of vaccines administered on days 0, 28, and 180. An accelerated schedule would be of great advantage to Japanese travelers, who could complete the required three doses for primary immunization in 1 month. [ABSTRACT FROM AUTHOR]

The reproductive biology of female Pasiphaea japonica, including the minimum size of females carrying embryos, spawning and hatching seasons, and brood size, was examined in Toyama Bay, central Japan, during 2004 through 2007. Sampling was conducted onboard a commercial seiner and a fisheries research vessel. The minimum size of ovigerous females was 47.7 mm body length. The percent occurrence of ovigerous females increased with body size, reaching 50% at 57.9 mm body length. Ovigerous females occurred all year round with a seasonal change in percent occurrence, ranging between 41.5 and 94.3% of the total observed females. Consequently, reproduction in females seems to occur throughout the year in Toyama Bay, with seasonal differences in reproductive intensity. Spawning intensity was high during April–July and October–January based on the occurrence of the early developmental stage of embryos, while hatching intensity was high in October–May based on the occurrence of embryos just before hatching. Brood sizes were similar at the same size of body length regardless of embryo stage, except just after spawning. [ABSTRACT FROM AUTHOR]

Abstract: The Japanese Silky chicken (SK) shows dermal and visceral hyperpigmentation. This study characterizes ultrastructurally the melanin granules developing in dermal melanocytes of the dorsal skin of SK, in an attempt to better understand the processes of melanogenesis in these permanently ectopic cells. The steps of melanogenesis are similar to those described for epidermal melanocytes, with melanosomes going from stage I to IV but, in SK, the maturation occurs in the cell body, as well as in the cytoplasmic processes. At stage III, the deposition of melanin is cumulative and can aggregate in rounded structures, which combine to turn into the mature granule. The final destiny of mature melanosomes is still unclear, although it was observed that dermal macrophages can accumulate melanin granules in their phagosomes. Even with the close proximity between melanocytes and other dermal cells, the transference of melanosomes was not observed. Our findings indicate that melanogenesis in dermal melanocytes in SK has the same morphological characteristics found in epidermal melanocytes, but the functional aspect still remains to be elucidated. [Copyright &y& Elsevier]

It is well known that heavy oil (HO) on the sea surface causes serious problems in the aquatic environment. In particular, some species of teleosts which develop on the sea surface are thought to be affected by the HO which flows out from tankers or coastal industry. However, the toxicological effects of HO are not fully understood. We performed exposure experiments using the Pleuronectiformean fish, spotted halibut (Verasper variegatus), which is an important fishery resource in Japan. In course of the development, HO-exposed embryos showed remarkable delay in developmental processes including somite formation. We further observed abnormal development of the head morphology. Notably, treated embryos had relatively small eyes and craniofacial structures. These findings strongly suggest that HO seriously affects the cell proliferation and differentiation of the embryo. In addition, HO-exposed embryos showed abnormal neuronal development. We also performed the exposure in the larval stage. Treatment of post-hatching larvae with HO resulted in significantly greater mortality compared with controls. Through these observations, we finally conclude that HO is strongly toxic to halibut in their early life stages. [Copyright &y& Elsevier]

We present a series of monozygous multiple gestations achieved following in vitro fertilization (IVF): one case of monochorionic triplet pregnancy and six cases of dizygotic triplet pregnancy. From September 2000 to December 2006, all patients achieving clinical pregnancy by ART were reviewed (n = 2433). A 37 year-old woman who delivered a healthy singleton after IVF returned two years later for FET, and a single blastocyst was transferred. This also resulted in pregnancy, but TV-USG revealed a single gestational sac with three distinct amniotic sacs, each containing a distinct fetal pole with cardiac activity. This pregnancy was electively terminated at nine weeks' gestation. An additional six cases of dizygotic triplets established after fresh embryo transfer (no ICSI or assisted hatching) are also described. Of these, one resulted in a miscarriage at eight weeks' gestation and five patients have an ongoing pregnancy. This case series suggests the incidence of dizygotic/monochorionic triplets following IVF is approximately 10 times higher than the expected rate in unassisted conceptions, and underscores the importance of a conservative approach to lower the number of embryos at transfer. The role of embryo transfer technique and in vitro culture media in the twinning process requires further study. [ABSTRACT FROM AUTHOR]

In an investigation of seed germination in Cardiocrinum cordatum var. glehnii, embryos in fresh seeds in October were underdeveloped and did not grow until September of the following year. Then, they grew rapidly and had fully elongated by early November. In the second spring after dispersal, radicles emerged under snow in late March and after snowmelt in April. Cotyledons emerged soon after radicles. In several laboratory experiments, embryos grew at 15°/5°C (light 12 h/dark 12 h) following 25°/15°C. Radicles emerged from seeds with fully elongated embryos at 5°–15°C after cold stratification at 0°–5°C. Cotyledons emerged in 2 wk from seeds with a radicle at 15°/5°C to 30°/20°C. Although seeds require c. 18–19 mo after dispersal to germinate in nature, under controlled conditions, they required only 9 mo with a sequence of 25°/15°C → 15°/5°C → 0°–5°C → 15°/5°C. This is practical knowledge for propagation of plants from seeds. GA3 treatment partially substituted for the high temperature requirement. Based on dormancy-breaking requirements, the seeds have deep simple morphophysiological dormancy (MPD). A literature review of seed dormancy in taxa of Liliaceae s. str. showed that phylogenetic position in this case is not a good predictor of level of MPD. [ABSTRACT FROM AUTHOR]

The article presents information on a research study regarding changes in seawater tolerance during the development of eyed-stage embryos in shishamo smelt Spirinchus lanceolatus. Shishamo smelt Spirinchus lanceolatus is an important resource on the Pacific coast of Hokkaido Prefecture in northern Japan and spawns in early winter. Four hundred adult female and male fish captured by purse seine at the mouth of the Mukawa River, southern Hokkaido, were used in this study. Eggs were spawned naturally on gravel.

Effects of two widely found chemical pollutants, bis(2-ethylhexyl) phthalate (DEHP) and benzo[a]pyrene (BaP), on the embryos of Japanese medaka were investigated. The embryos were exposed to different concentrations (0.01, 0.1, 1, and 10 μg/l) of DEHP and BaP. The following were investigated: (1) hatching time and hatching rate in embryos, (2) mortality, sex ratio, body weight and gonadosomatic index (GSI) in adulthood. These two chemicals delayed the hatching time without dose-dependence, but these chemicals had no effect on hatching rate. Mortality was raised and body weight was reduced by DEHP and BaP-treatment; distortion of sex ratio appeared at the lowest concentration of DEHP tested. GSI was decreased because of the BaP-treatment. DEHP and BaP negatively affected Japanease medaka embryos, and the influences of the effects continued into adult stage. Moreover, the effects did not appear to be necessarily dose-dependent. [Copyright &y& Elsevier]

To support the recovery of Morus boninensis Koidz. (Moraceae), a critically endangered species distributed in the Bonin Islands of Japan, we need to distinguish between pure M. boninensis and hybrid (M. boninensis × M. acidosa Griff.) seedlings within populations derived from M. boninensis mother trees. Therefore, we developed three co-dominant SCAR (Sequence Characterized Amplified Region) markers to distinguish between M. boninensis, M. acidosa and their hybrids. All three markers could clearly identify the species and the hybrids, although we observed one case of a putative null allele or a backcross at one locus (Mos0251-1). Hence, we suggest that pure and hybrid seedlings could be unambiguously identified by the simultaneous use of the three markers. When we investigated a seedling population produced by a single mother tree growing in the Iguma-bay population of Haha-jima Island, the proportion of embryos formed by pollination from M. acidosa pollen was about 13%. All hybrid seedlings identified according to the molecular markers showed incisions in their leaves, a morphological character that has never been observed in pure M. boninensis individuals. Our results suggest that M. boninensis can potentially be multiply by excluding hybrids from the seedling pools, using incisions on the leaves as morphological markers to identify them. [ABSTRACT FROM AUTHOR]

Examines the effect of cryoprotectant agents on the survival and in vitro development of hamster embryos in Tokyo, Japan. Retardation of one-cell embryo development after exposure to agents; Comparison of cryoprotectant agent tolerance between two- and eight-cell embryo; Recommendations for successful vitrification of preimplantation embryos.

Sex ratio and breeding habits of Planaxis sulcatus were studied for Japanese and other Indo-Pacific populations. On the coast of the Kii Peninsula, Japan, the gonad of P. sulcatus is developed from June through August, when pairing of the snails is observed. The females contain embryos in their brood pouches and release veligers. New recruits appear in autumn, grow during the next spring and summer, and merge with the older-year cohort in size in early autumn. The position on the shore of the first-year and the older-year snails is lower in spring and early summer than in other seasons. Sex ratio was significantly different between populations, seemingly due to different percentages of large snails which were mostly females. Inspection of the samples from a wider geographic range, from Japan to India, detected various sex ratios among the localities; females were dominant in the samples composed exclusively of large individuals. The most developed embryos in the female brood pouches were veligers. Parthenogenetic development and rearing of embryos up to crawling juveniles, which have been reported for the populations in the inner Arabian Sea, does not seem to hold for these other Indo-Pacific populations. [ABSTRACT FROM AUTHOR]

The Great Eastern Japan Earthquake of 2011 and consequent tsunami affected many IVF clinics in northeastern Japan. Twelve clinics lost at least one embryo, but all the frozen embryos stored in liquid nitrogen tanks were safe. Emergency power supply is the first priority issue in an unpredictable natural disaster. [ABSTRACT FROM AUTHOR]

One of the possible countermeasures for pollinosis caused by sugi (Cryptomeria japonica), a serious public health problem in Japan, is the use of male sterile plants (MSPs; pollen-free plants). However, the production efficiencies of MSPs raised by conventional methods are extremely poor, time consuming, and resulting in a high seedling cost. Here, we report the development of a novel technique for efficient production of MSPs, which combines marker-assisted selection (MAS) and somatic embryogenesis (SE). SE from four full sib seed families of sugi, carrying the male sterility gene MS1, was initiated using megagametophyte explants that originated from four seed collections taken at one-week intervals during the month of July 2017. Embryogenic cell lines (ECLs) were achieved in all families, with initiation rates varying from 0.6% to 59%. Somatic embryos were produced from genetic marker-selected male sterile ECLs on medium containing maltose, abscisic acid (ABA), polyethylene glycol (PEG), and activated charcoal (AC). Subsequently, high frequencies of germination and plant conversion (≥76%) were obtained on plant growth regulator-free medium. Regenerated plantlets were acclimatized successfully, and the initial growth of male sterile somatic plants was monitored in the field. [ABSTRACT FROM AUTHOR]