Influence of DNA extraction kits on freshwater fungal DNA metabarcoding

[Background] Environmental DNA (eDNA) metabarcoding is a common technique for efficient biodiversity monitoring, especially of microbes. Recently, the usefulness of aquatic eDNA in monitoring the diversity of both terrestrial and aquatic fungi has been suggested. In eDNA studies, different experimental factors, such as DNA extraction kits or methods, can affect the subsequent analyses and the results of DNA metabarcoding. However, few methodological studies have been carried out on eDNA of fungi, and little is known about how experimental procedures can affect the results of biodiversity analysis. In this study, we focused on the effect of DNA extraction method on fungal DNA metabarcoding using freshwater samples obtained from rivers and lakes. [Methods] DNA was extracted from freshwater samples using the DNeasy PowerSoil kit, which is mainly used to extractmicrobial DNA from soil, and the DNeasy Blood & Tissue kit, which is commonly used for eDNA studies on animals. We then compared PCR inhibition and fungal DNA metabarcoding results; i.e., operational taxonomic unit (OTU) number and composition of the extracted samples. [Results] No PCR inhibition was detected in any of the samples, and no significant differences in the number of OTUs and OTU compositions were detected between the samples processed using different kits. These results indicate that both DNA extraction kits may provide similar diversity results for the river and lake samples evaluated in this study. Therefore, it may be possible to evaluate the diversity of fungi using a unified experimental method, even with samples obtained for diversity studies on other taxa such as those of animals

Oral human cytomegalovirus prevalence and its relationships with periodontitis and Porphyromonas gingivalis in Japanese adults: a cross-sectional study

Objective: This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology: In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results: HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions: Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease

Noninvasive Positive Pressure Ventilation against Reperfusion Pulmonary Edema following Percutaneous Transluminal Pulmonary Angioplasty

A 69-year-old man with chronic thromboembolic pulmonary hypertension (CTEPH) was on amblatory oxygen inhalation therapy (3 L/min) and scheduled for percutaneous transluminal pulmonary angioplasty (PTPA). The patient's New York Heart Association functional status was class III with recent worsening of dyspnea and apparent leg edema. Transthoracic echocardiography revealed right ventricular enlargement with mean pulmonary artery pressure of 42 mmHg. After PTPA, he was complicated with postoperative reperfusion pulmonary edema, and noninvasive positive pressure ventilation (NPPV) was applied immediately. Hypoxemia was successfully treated with 15 days of NPPV. Although mean pulmonary artery pressure was unchanged, his brain natriuretic peptide level decreased from preoperative 390.3 to postoperative 44.3 pg/dL. In addition, total pulmonary resistance decreased from preoperative 18 to postoperative 9.6 wood unit·m2. The patient was discharged on day 25 with SpO2 of 95% on 5 L/min of oxygen inhalation. Because pulmonary edema is a postsurgical life-threatening complication following PTPA, application of NPPV should be considered

The intertidal macrobenthic fauna of the Hatakejima Experimental Field, Wakayama Prefecture, Japan, in 2019

ファイル差し替え（2021-05-17）Hatakejima Experimental Field is located in Tanabe Bay, Wakayama Prefecture, Japan, which is composed of Hatakejima Island and Komarujima Islet, connected to the former in low tide. Hatakejima Island was purchased by Kyoto University and was designated as the “Hatakejima Experimental Field” in 1968. The year 2019 marks the 50th year of the long-term surveys that have been formally conducted on the experimental field since 1969 (i.e., the Century of Research Project). We conducted a field survey to record the macrobenthic fauna of the experimental field in 2019. A total of 168 species of 11 phyla were recorded in this survey. In each phylum, the number of species is listed as follows in descending order: Mollusca (78 spp.), Arthropoda (27 spp.), Echinodermata (23 spp.), Annelida (21 spp.), Cnidaria (7 spp.), Porifera (3 spp.), Nemertea (3 spp.), Platyhelminthes (2 spp.), Chordata (2 spp.), Bryozoa (1 sp.), and Hemichordata (1 sp.). We also recorded and discussed the influence of recent environmental changes around the Hatakejima Experimental Field. Tropical sea urchin species disappeared in the winter of 2017–2018 following the large meander of the Kuroshio Current, which led to decreasing water temperatures. The population of the seagrass Zostera japonica drastically decreased on the western sandy shore of the island in 2019, most likely because of two big typhoons in September 2018. We must conduct continuous observations to aid the recovery of seagrass-associated communities and protect the experimental field to keep high biodiversity of macrobenthic fauna in the future

フォトダイオードアレイ検出HPLC を用いた微分クロマトグラフ法によるカフェインの定量

We assessed caffeine levels in bathwater and diluted urine samples by derivative processing of chromatograms obtained through the use of HPLC with photodiode array detection. Chromatograms were differentiated on the basis of d（Abs）/d（min）．The peak position of the target analyte, caffeine, was observed to shift depending on the derivative order applied, but was constant when applying the same derivative order, such that retention times were constant even after differentiation. When the method detection limit was examined using standard solutions, the varying absorption of acetonitrile during a linear gradient elution from 1 to 100 % acetonitrile was shown to have an effect, such that the linearity of the peak height deteriorated if caffeine was present at low concentrations. Recovery of caffeine from real samples was optimized in the case of both linear gradient elution and isocratic elution（ 10 % acetonitrile）when applying the second derivative and using a detection wavelength of 259 nm. Interestingly, this does not correspond to the maximum absorption of caffeine. Thus the optimum wavelength for use with this differential chromatographic method is one that is not affected by the absorbance of the eluent and at which the target analyte shows significant absorbance, even if it is not the maximum absorbance. Such conditions were found to be necessary for the high-sensitivity measurement of caffeine. It is believed that this method is applicable to the HPLC analysis of compounds other than caffeine and this technique is expected to be effective when applied to an automated analyzer and should be helpful to analysts unfamiliar with HPLC.フォトダイオードアレイ検出HPLC を用いた微分クロマトグラフ法による希釈尿あるいは風呂水からのカフェインの回収について検討した。通常のクロマトグラムを微分（d（Abs）/d（min））し，微分クロマトグラムを得る。目的の化合物のピーク位置は，微分次数によるずれるが，同じ微分次数で比較すると一定しており，保持時間による定性能は微分後も維持されていた。標準溶液を用いた定量下限値は，リニアグラジエント溶出（1 → 100 ％アセトニトリル）では溶離液として用いたアセトニトリルの吸収が影響を与え，カフェインが低濃度の場合にはピーク高さとの直線性が悪化した。実試料からのカフェインの回収は，波長259 nm，2 次微分実行時にリニアグラジエント溶出，アイソクラティック溶出（10 ％アセトニトリル）共最適であった。この条件は必ずしもカフェインが極大吸収を持つ波長，微分次数ではない。本結果は微分クロマトグラフ法での検出条件として，1）溶離液吸収の影響を受け難い波長を選択すること，2）目的物質であるカフェインの吸光度が極大でなくてもある程度大きいことが高感度測定に必要であることを示す。今回の結果はカフェインのみならず，他の化合物をHPLC 分析する場合，特にHPLC に不慣れな使用者が条件検討する場合や自動分析装置に応用する場合にも利用可能な知見であると考えられる

Risk Factors for Infection in Patients with Remitted Rheumatic Diseases Treated with Glucocorticoids

It is well known that infection is one of the major causes of morbidity and mortality in rheumatic disease patients treated with high-dose glucocorticoids, especially in the early phase after achievement of disease remission. The aim of this study was to identify the risk factors for infection, with a focus on the dose of glucocorticoids administered, following the achievement of disease remission in rheumatic diseases patients. We retrospectively analyzed the medical records of rheumatic disease patients who had been treated with glucocorticoids. The primary endpoint was the incidence rate of infection during a period from 1 to 2 months after the commencement of treatment. From April 2006 to March 2010, 19 of 92 patients suffered from infection during the observation period. Age≧65 yrs, presence of interstitial pneumonia, diagnosis of systemic vasculitis and serum creatinine level≧2.0mg/dl were found to be univariate predictors for infection. However, only the presence of interstitial pneumonia was an independent risk factor for infection (HR＝4.50, 95%CI＝1.65 to 14.44) by the Cox proportional hazard model. Even after achievement of clinical remission, careful observation is needed for patients with interstitial pneumonia, more so than for those receiving high-dose glucocorticoids

Association of blood pressure and renal outcome in patients with chronic kidney disease; a post hoc analysis of FROM-J study

It is well-known that hypertension exacerbates chronic kidney disease (CKD) progression, however, the optimal target blood pressure (BP) level in patients with CKD remains unclear. This study aimed to assess the optimal BP level for preventing CKD progression. The risk of renal outcome among different BP categories at baseline as well as 1 year after, were evaluated using individual CKD patient data aged between 40 and 74 years from FROM-J [Frontier of Renal Outcome Modifications in Japan] study. The renal outcome was defined as >= 40% reduction in estimated glomerular filtration rate to130 mmHg group. A significant increase in the renal outcome was found only in the group of diastolic BP >= 90 mmHg. The group of BP= 130 mmHg at baseline. Targeting SBP level<130 mmHg would be associated with the preferable renal outcome.Clinical Trial Registration-URL: https://www.umin.ac.jp/ctr/. Unique identifier: UMIN000001159 (16/05/2008)

Role of PERK in mitochondrial function

Mitochondria play a central role in the function of brown adipocytes (BAs). Although mitochondrial biogenesis, which is indispensable for thermogenesis, is regulated by coordination between nuclear DNA transcription and mitochondrial DNA transcription, the molecular mechanisms of mitochondrial development during BA differentiation are largely unknown. Here, we show the importance of the ER-resident sensor PKR-like ER kinase (PERK) in the mitochondrial thermogenesis of brown adipose tissue. During BA differentiation, PERK is physiologically phosphorylated independently of the ER stress. This PERK phosphorylation induces transcriptional activation by GA-binding protein transcription factor α subunit (GABPα), which is required for mitochondrial inner membrane protein biogenesis, and this novel role of PERK is involved in maintaining the body temperatures of mice during cold exposure. Our findings demonstrate that mitochondrial development regulated by the PERK–GABPα axis is indispensable for thermogenesis in brown adipose tissue