Immunoelectron microscopy using chromogranin A-specific antibodies has been proposed as an efficient technique for identification of secretory granules (SGs) in tumor cells with evidence of apparent ...neuroendocrine differentiation. Using an antigen retrieval (AR) method, we succeeded in immunolabeling SGs with antibodies in ultrathin sections of routinely processed epoxy-embedded blocks of tissue. Samples of an insulinoma were fixed in 2% glutaraldehyde, postfixed in 1% OsO4, and embedded in epoxy resin. Ultrathin sections were immunostained with chromogranin A-specific antibodies and gold-conjugated second antibodies. There was no significant labeling in the absence of AR. Neither etching with sodium metaperiodate nor microwave irradiation of ultrathin sections in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0) was effective in improving the efficiency of immunolabeling. However, ultrathin epoxy-embedded sections that were microwaved in alkaline solution (pH 10) were adequately labeled (5.2 ± 0.34 particles per SG). Moreover, considerably improved efficiency of immunostaining was achieved by microwaving sections in alkaline solution (pH 10) with subsequent immunostaining at 60C (12.2 ± 0.51 particles per SG). This method can also be applied to epoxy-embedded sections obtained from formalin-fixed, paraffin-embedded blocks of tissue and was even valid for an old epoxy-embedded block of tissue prepared 15 years previously.
Despite recent advances in sequencing technology and large-scale drug screenings employing hundreds of cell lines, the predictive accuracy of mutation-based biomarkers is still insufficient as a ...guide for cancer therapy. Therefore, novel types of diagnostic methods using alternative biomarkers would be highly desirable. We have hypothesized that sensitivity-specific changes in the phosphorylation of signaling molecules could be useful in this respect. Here, with the aim of developing a method for predicting the response of cancers to cisplatin using a combination of specific biomarker(s) and patient-derived tumor organoids (PDOs), we found that cisplatin-sensitive cell lines or PDOs showed enhanced phosphorylation of c-Jun (p-c-Jun) within 24 h after cisplatin treatment. We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy (docetaxel/cisplatin/5-fluorouracil) in 6 matched patients. Mechanistically, the c-Jun induction was partly related to TNF signaling induced by cisplatin. Our data suggest that enhanced phosphorylation of c-Jun in response to cisplatin treatment could be a predictive biomarker for the efficacy of cisplatin in selected cancer patients.
This study shows the potential application of c-Jun phosphorylation upon cisplatin treatment as a biomarker for cisplatin sensitivity in combination with patient-derived tumor organoids (PDOs). We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy in 6 matched patients with esophageal squamous cell carcinoma.
Ixodid ticks were collected from medium-sized to large mammals in Ehime Prefecture, Shikoku, Japan. Ten species of ticks (Amblyomma testudinarium, Dermacentor taiwanensis, Haemaphysalis flava, H. ...formosensis, H. hystricis, H. longicornis, H. megaspinosa, Ixodes nipponensis, I. ovatus, and I. tanuki) were collected from a total of 29 mammals comprising 11 species. Haemaphysalis hystricis, a possible vector of Japanese spotted fever in Ehime prefecture, was collected from Canis lupus familiaris (domestic dog), Martes melampus melampus, and Sus scrofa leucomystax. This is a first report of H. hystricis from the domesticated dog in the endemic area of Japanese spotted fever. This suggests that it is necessary to pay attention to dogs as a host of the vector ticks for Japanese spotted fever control. Nyctereutes procyonoides and Ma. melampus are new hosts for A. testudinarium. Nyctereutes procyonoides, Mustela itatsi, and Lepus brachyurus are new hosts for H. formosensis. Martes melampus is a new host for H. hystricis.
Organic amendments are important sources of nitrous oxide (N
O) emissions from agricultural soils. In 2020, the total amount of N in organic amendments applied to Japanese agricultural soils (440 ...ktN) was larger than that of synthetic fertilizer (374 ktN). However, N
O emissions from organic amendments were estimated by using the country-specific N
O emission factor (EF) for synthetic fertilizer (0.31 % for rice paddy, 2.9 % for tea, and 0.62 % for other crops) in the National Greenhouse Gas Inventory Report of Japan. Thus, we conducted a N
O flux measurement campaign at 12 different experimental sites across Japan to estimate fertilizer-induced N
O EFs for major organic amendments in Japan, that is, poultry manure compost, swine manure compost, cattle manure compost, and organic fertilizer pellets. In addition, we conducted systematic review of N
O emissions and EFs for organic amendments, including data from our measurement campaign and published data from peer-reviewed papers in Japan. The final dataset, including the field measurement campaign and published data, resulted in 404 observations (including synthetic fertilizer and zero-N control) in 29 sites. Results showed that soil type affected EFs, that is, the mean EF of Andosols was lower than that of non-Andosols, which is similar to the case of EFs for synthetic fertilizer. Mean EFs for poultry manure compost, swine manure compost, cattle manure (compost and slurry), and non-animal manure organic fertilizers were 0.83 % (uncertainty range of 2.5th and 97.5th percentile: 0.09 % to 3.46 %), 0.70 % (0.02 % to 2.45 %), 0.39 % (0.00 % to 1.62 %), and 1.16 % (0.41 % to 3.03 %), respectively, when weighted by area of soil types. The mean EF of all organic amendments was 0.84 % (0.00 % to 2.91 %), when the area of soil type and amount of organic amendment used in Japan were considered. Our study provides country-specific EFs to estimate N
O emission from organic amendments in Japan.
Here, we describe the reproductive behaviour and life history traits of Acanthosoma firmatum (Walker, 1868) (Heteroptera: Acanthosomatidae). Females laid eggs in tight masses on the underside of ...leaves of the host plant and the clutch size often exceeded 130, which seems to be the largest among those recorded from Acanthosoma and the related Sastragala species. Females stayed on the clutches in a straddling posture. When disturbed, females showed aggressive responses, such as tilting the body towards the source of disturbance. No female was observed tending to the second instar nymphs that had left the natal leaf. The females sometimes copulated with males during maternal care.
Despite recent advances in sequencing technology and large-scale drug screenings employing hundreds of cell lines, the predictive accuracy of mutation-based biomarkers is still insufficient as a ...guide for cancer therapy. Therefore, novel types of diagnostic methods using alternative biomarkers would be highly desirable. We have hypothesized that sensitivity-specific changes in the phosphorylation of signaling molecules could be useful in this respect. Here, with the aim of developing a method for predicting the response of cancers to cisplatin using a combination of specific biomarker(s) and patient-derived tumor organoids (PDOs), we found that cisplatin-sensitive cell lines or PDOs showed enhanced phosphorylation of c-Jun (p-c-Jun) within 24 h after cisplatin treatment. We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy (docetaxel/cisplatin/5-fluorouracil) in 6 matched patients. Mechanistically, the c-Jun induction was partly related to TNF signaling induced by cisplatin. Our data suggest that enhanced phosphorylation of c-Jun in response to cisplatin treatment could be a predictive biomarker for the efficacy of cisplatin in selected cancer patients. This study shows the potential application of c-Jun phosphorylation upon cisplatin treatment as a biomarker for cisplatin sensitivity in combination with patient-derived tumor organoids (PDOs). We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy in 6 matched patients with esophageal squamous cell carcinoma.
