Precipitant-Free Crystallization of Lysozyme and Glucose Isomerase by Drying

Protein crystallization is usually conducted by using precipitants, although the “salting-out” phenomenon is still unclear and complex. Moreover, the addition of precipitants sometimes results in irreversible disordered precipitation of protein molecules. Although precipitant-free lysozyme crystals obtained by centrifugal concentration showed significant changes in three-dimensional structure compared to the structure of salted-out crystals, it was rather difficult to mount crystals from a viscous dense liquid phase after centrifugal concentration, and the quality of the crystals often deteriorated during the mounting process. Here we present novel precipitant-free crystallization methods, which were effective for lysozyme and glucose isomerase. Tetragonal lysozyme crystals were successfully crystallized in a glass capillary simply by drying highly concentrated lysozyme solution in the presence of 0.01 M hydrochloric acid without using any precipitants. Glucose isomerase dissolved in ultra-pure water was also successfully crystallized in hanging drops by drying highly concentrated solution under low-humidity conditions. Oscillation images of the obtained crystals were safely collected without handling; they clearly indicated the crystals had a tetragonal form for lysozyme and an orthorhombic form for glucose isomerase, and their lattice parameters are similar to those of previously reported crystals obtained by salting-out methods

Transesterification of Lignocellulosic Biomass Using Ionic Liquids

13301甲第4957号博士（工学）金沢大学博士論文要旨Abstract 要約Outline 掲載予定あ

Evaluation of the Effect of Culling on Browse Damage by the Japanese Serow in Gifu Prefecture, Japan

The Japanese serow (Capricornis crispus; serow) is a protected territorial ungulate native to Japan. However, locally overabundant serow populations can damage forest plantations and agriculture through browsing. Despite government permitted annual culling of serows on forest lands of Gifu Prefecture, Japan, browse damage continues to be reported in hinoki cypress (Chamaecyparis obtusa; cypress) plantations. Sika deer (Cervus nippon; deer), which are co-located with serows can also browse cypress, but their impacts have never been evaluated. The objective of our research was to evaluate the involvement of each species in browse damage and to establish the damage-causing mechanisms after serow culling at selected study sites (T1 [0.3 ha], T2 [0.2 ha], and T3 [1.1 ha]) in 3 cypress plantations in Takayama City, Gifu Prefecture, where serow culling was conducted. In 2019 and 2020, 2 and 2 serows were culled in T1, 3 and 0 in T2, and 1 and 1 in T3, respectively. Forestry workers also applied a chemical repellent (ziram-based fungicide) to some stands in T3 in October 2019 and May 2020. Between December 2018 and September 2020, we used camera traps to monitor activity patterns of serows and deer and the replacement of territorial serows before and after culling. We also investigated seasonal browsing impacts between August 2019 and June 2020 by thoroughly checking for browsing marks on the terminal shoot. Serows and deer accounted for 79% and 21% of camera-trap videos, respectively. Despite annual culling, serows were recorded at all browsed sites before the next growing season. Browse damage was higher in autumn and winter, but in T3 it was reduced when the repellent was applied. Management of ungulate browse damage to cypress will require accurate identification of species causing the damage, monitoring serow activity before and after culling, and a using repellent immediately before browsing seasons

A crowdsourcing approach to collecting photo-based insect and plant observation records

Scientific field observation by members of the public is known as citizen science and has become popular all across the world. Citizen science is advantageous for collecting large amounts of scientific data and can be seen as a crowdsourcing approach to data collection. Information and communications technology is enhancing the availability of citizen science. Mobile devices, such as mobile phones, that have a digital camera with a global positioning system (GPS) are necessities for contemporary life and can be utilised as powerful observation tools in citizen science. A web-based system has been developed as a data collection tool for citizen science. Participants submit an e-mail with a photo taken by their mobile phones. The photos contain location information, which can be easily and automatically embedded if the mobile phone is equipped with GPS. Collaboration has been undertaken with regional event managers, such as museum curators and held citizen science events in each region and for various target taxonomic groups. All photos were stored in the data server and the organisms were taxonomically identified by citizen scientists, regional managers and the authors. In total, 154 species and 843 data records were collected in this project conducted from 2011 to 2016

Hoxa13 regulates expression of common Hox target genes involved in cartilage development to coordinate the expansion of the autopodal anlage

To elucidate the role of Hox genes in limb cartilage development, we identified the target genes of HOXA11 and HOXA13 by ChIP‐Seq. The ChIP DNA fragment contained evolutionarily conserved sequences and multiple highly conserved HOX binding sites. A substantial portion of the HOXA11 ChIP fragment overlapped with the HOXA13 ChIP fragment indicating that both factors share common targets. Deletion of the target regions neighboring Bmp2 or Tshz2 reduced their expression in the autopod suggesting that they function as the limb bud‐specific enhancers. We identified the Hox downstream genes as exhibiting expression changes in the Hoxa13 knock out (KO) and Hoxd11‐13 deletion double mutant (Hox13 dKO) autopod by Genechip analysis. The Hox downstream genes neighboring the ChIP fragment were defined as the direct targets of Hox. We analyzed the spatial expression pattern of the Hox target genes that encode two different categories of transcription factors during autopod development and Hox13dKO limb bud. (a) Bcl11a, encoding a repressor of cartilage differentiation, was expressed in the E11.5 autopod and was substantially reduced in the Hox13dKO. (b) The transcription factors Aff3, Bnc2, Nfib and Runx1t1 were expressed in the zeugopodal cartilage but not in the autopod due to the repressive or relatively weak transcriptional activity of Hox13 at E11.5. Interestingly, the expression of these genes was later observed in the autopodal cartilage at E12.5. These results indicate that Hox13 transiently suspends the cartilage differentiation in the autopodal anlage via multiple pathways until establishing the paddle‐shaped structure required to generate five digits

Paraganglioma that caused sinus arrest

Paragangliomas are neural-crest-derived nonepithelial neuroendocrine tumors distributed along the parasympathetic and sympathetic nerves. To our knowledge, no studies were reported regarding sinus arrest on day 4 after paraganglioma resection. A 66-year-old female patient with a history of pulmonary vein isolation visited our department for sigmoid colon cancer treatment. Enhanced computed tomography revealed an enhanced small nodule-like lymph node near the root of the inferior mesenteric artery. The patient underwent laparoscopic colectomy with regional lymph node dissection. Postoperatively, paroxysmal atrial fibrillation attacks developed, and the patient resumed oral medication. Additionally, sinus arrest after tachycardia developed. Changing the oral medication could maintain her circulatory dynamics. Pathological examination revealed that differentiated tubular adenocarcinoma infiltrated the submucosa. Immunohistochemically, the excised nodule as a lymph node was considered a functional paraganglioma. Our case indicates that paraganglioma resection and oral medication resumption may contribute to sinus arrest. When arrhythmias affecting the circulation occur perioperatively, the presence of a catecholamine-producing tumor should be considered in addition to cardiac disease

Pericardial Mesothelioma in a Dog: The Feasibility of Ultrasonography in Monitoring Tumor Progression

A 6-year-old neutered male Yorkshire Terrier presented with recurrent pericardial effusion. Although clinical examinations including computed tomography were inconclusive, an exploratory thoracotomy revealed multiple small nodules and plaques on the inner surface of the pericardial sac (Day 1). A subtotal pericardiectomy was performed to prevent cardiac tamponade due to the increasing pericardial effusion, and the resected section of the pericardium was histopathologically diagnosed with mesothelioma. After surgery, chemotherapy with intrathoracic carboplatin was commenced. During the course of the treatment, a detailed follow-up ultrasonographic scan was performed to detect early lesions disseminated on the pleura, originating from the primary pericardial mesothelioma. On Day 101, the minute pleural nodules, which were disseminated lesions as predicted, were successfully imaged by ultrasonography. As the clinical stage advanced, the nodules were observed to gradually increase in size and number, implying tumor progression. These observations highlight the feasibility of ultrasonography in detecting minute disseminated lesions at an early stage, monitoring tumor progression, and thereby, predicting the prognosis of canine pericardial mesothelioma

パラオで採集した海洋糸状菌TUF98F139株が生産する微少管重合阻害活性物質

東京水産大学海洋環境学科東京水産大学海洋環境学科東京水産大学海洋環境学科東京大学分子細胞生物学研究所東京水産大学練習船海鷹丸東京水産大学練習船海鷹

Metastatic Renal Cell Carcinoma to the Left Sphenoid Sinus: A Case Report in Light of the Literature

A 79-year-old Japanese woman presented with a rare case of metastatic renal cell carcinoma to the left sphenoid sinus with left nasal bleeding. She had previously had right radical nephrectomy for renal cell carcinoma at the age of 64 years and brain and spinal cord infarction at 74 years. Endoscopic examination revealed no mass in the nasal cavity. CT and MRI revealed a tumor in the left sphenoid sinus. The size of the tumor increased gradually from 12 to 15 years after the radical nephrectomy. Complete resection with endoscopic surgery was performed without preoperative embolization. The tumor cells had clear cytoplasm and were arranged in a trabecular pattern lined by a layer of endothelial cells. These findings were identical to the pathological findings of the surgical specimen of the renal cell carcinoma from 15 years previous. A pathological diagnosis of metastatic renal cell carcinoma of clear cell type (grade 1) was made. PET-CT demonstrated no metastasis. The patient’s condition was successfully managed with excision of the tumor, and she remains well with no evidence of recurrence and metastasis 36 months after treatment. Metastatic renal cell carcinoma to the sphenoid sinus is rare, but it might be considered in the differential diagnosis of masses in the paranasal sinus even long after initial treatment of renal cancer