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More specific screening systems for cervical cancer may become necessary as the human papillomavirus (HPV) vaccine becomes more widespread. Although p16/Ki-67 dual-staining cytology has several advantages, it requires advanced diagnostic skills. Here, we developed an automated on-chip immunostaining method using a microfluidic device. An electroactive microwell array (EMA) microfluidic device with patterned thin-film electrodes at the bottom of each microwell was used for single-cell capture by dielectrophoresis. Immunostaining and dual staining for p16/Ki-67 were performed on diagnosed liquid cytology samples using the EMA device. The numbers of p16/Ki-67 dual-stained cells captured by the EMA device were determined and compared among the cervical intraepithelial neoplasia (CIN) lesion samples. Seven normal, fifteen CIN grade 3, and seven CIN grade 2 samples were examined. The percentage of dual-positive cells was 18.6% in the CIN grade 2 samples and 23.6% in the CIN grade 3 samples. The percentages of dual-positive staining increased significantly as the severity of the cervical lesions increased. p16/Ki67 dual immunostaining using the EMA device is as sensitive as the conventional method of confirming the histopathological diagnosis of cervical samples. This system enables a quantified parallel analysis at the individual cell level., (© 2023. Springer Nature Limited.)

Background: To treat diseases caused by genetic variants, it is necessary to identify disease-causing variants in patients. However, since there are a large number of disease-causing variants, the application of AI is required. We propose AI to solve this problem and report the results of its application in identifying disease-causing variants., Methods: To assist physicians in their task of identifying disease-causing variants, we propose an explainable AI (XAI) that combines high estimation accuracy with explainability using a knowledge graph. We integrated databases for genomic medicine and constructed a large knowledge graph that was used to achieve the XAI., Results: We compared our XAI with random forests and decision trees., Conclusion: We propose an XAI that uses knowledge graphs for explanation. The proposed method achieves high estimation performance and explainability. This will support the promotion of genomic medicine.

Background: There is no standard therapy for hemodialysis (HD) patients with COVID-19. Data on remdesivir in HD patients with COVID-19 are scarce., Methods: We retrospectively analyzed 25 HD patients with COVID-19 treated with remdesivir., Results: The median age of the patients was 78 years (range, 45-92 years) and was predominantly male (84%). A total of 44% of the patients had mild disease, 36% had moderate-1, and 20% had moderate-2. The most common symptoms were fever (76%) and coughing (44%). The most common comorbidity was renal failure (100%), followed by hypertension (60%) and cardiac disease (44%). The most frequent biomarker was elevated creatinine (100%), followed by C-reactive protein (80%), lymphopenia (76%), and D-dimer (68%). C-reactive protein levels decreased significantly before and after remdesivir administration (p < 0.001). Two patients showed deterioration, but none died. All patients recovered from COVID-19 and no adverse effects of treatment with remdesivir were observed., Conclusion: Our study suggests the safe use of remdesivir in HD patients with COVID-19., (© 2022 Sociedad Española de Nefrología. Published by Elsevier España, S.L.U.)

Introduction: We investigated the association between stroke and asymptomatic latent syphilis (ALS) in geriatric patients., Methods: This retrospective observational study included patients aged >65 years who underwent routine qualitative rapid plasma reagin (RPR) and Treponema pallidum Latex Agglutination (TPLA) tests at a 161-bed community acute care hospital with long-term care facilities in Kamakura, Japan, from August 2014 to February 2019. Asymptomatic patients with a positive TPLA test result were diagnosed with ALS. Ninety-six patients with ALS were included in the study. Fifty-one patients (53.1%) had a positive RPR test. Comorbidities included hypertension (n = 44; 45.8%), chronic kidney disease (n = 44; 45.8%), and fracture (n = 29; 30.2%). No significant differences were found in sex, age, or comorbidities in univariate analyses. Multivariate analysis of the TPLA-positive geriatric patients revealed that a positive RPR test (odds ratio = 9.06; 95% confidence interval = 1.69-48.5; p = 0.01) was associated with a history of stroke., Conclusions: For geriatric patients with ALS, a positive qualitative RPR test was associated with a history of stroke. Medical evaluation and management of the risk factors for stroke are more necessary for ALS with a positive RPR qualitative test., (Copyright © 2021 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Several specific tests for cervical screening have been developed recently, including p16/Ki67 dual immunostaining for diagnosing high-risk human papillomavirus positive squamous intraepithelial lesion in the cervix. However, manual screening of cells in an entire glass slide is currently a standard clinical procedure for quantification and interpretation of immunocytochemical features of the cells. Here, we developed a microfluidic device containing an electroactive microwell array with barriers (EMAB) for highly efficient single-cell trapping followed by on-chip immunofluorescence analysis with minimum loss of the sample. EMAB utilizes patterned electrodes at the bottom of cell-sized microwells to trap single cells using dielectrophoresis (DEP) and cell-holding structures behind the microwells to stabilize the position of trapped cells even without DEP. Using the device, we evaluated the performance of p16/Ki67 dual immunostaining of HeLa cells on the chip. The device shows 98% cell-trapping efficiency as well as 92% cell-holding efficiency against the fixed HeLa cells, and we successfully demonstrated high-efficiency on-chip immunofluorescence analysis with minimal loss of sample. p16/Ki67 dual immunostaining using EMAB may be useful for complementary tests for cervical screening in confirming the histopathological diagnosis.

We report the case of a 32-year-old Japanese female who presented with a mild sore throat, rash and posterior cervical and posterior auricular lymphadenopathy. She was diagnosed with rubella. Japan has experienced multiple outbreaks of rubella in recent times. Travelers to Japan should consider getting vaccinated for rubella., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Detecting the bio-potential changes of plants would be useful for monitoring their growth and health in the field. A sensitive plant monitoring system with flexible boron-doped diamond (BDD) electrodes prepared from BDD powder and resin (Nafion or Vylon-KE1830) was investigated. The properties of the electrodes were compared with those of small BDD plate-type electrodes by monitoring the bioelectric potentials of potted Aloe and hybrid species in the genus Opuntia . While flexible BDD electrodes have wide potential windows, their cyclic voltammograms are different from those of the BDD plate. Further, the potential gap between a pair of electrodes attached to the plants changes as the plants are stimulated artificially with a finger touch, suggesting that the bioelectric potentials in the plant also changed, manifesting as changes in the potential gap between the electrodes. The BDD electrodes were assessed for their response reproducibility to a finger stimulus for 30 days. It was concluded that the plant monitoring system worked well with flexible BDD electrodes. Further, the electrodes were stable, and as reliable as the BDD plate electrodes in this study. Thus, a flexible and inexpensive BDD electrode system was successfully fabricated for monitoring the bioelectric potential changes in plants., Competing Interests: The authors declare no conflict of interest.

Background/purpose: Our aim was to describe the clinical features and prognostic factors of Gram-negative rod bacteremia (GNRB) after cardiovascular surgery (CVS)., Methods: This retrospective observational study included adults with GNRB onset within 100 days after CVS at a single institution from April 2004 to May 2013. Clinical data regarding episodes of GNRB were collected from patients' medical charts. Those having polymicrobial bacteremia with a bacterium other than a GNR were excluded., Results: Among 2017 CVS patients, GNRB occurred in 78. Klebsiella, Pseudomonas aeruginosa, Enterobacter, and Escherichia coli were the most commonly isolated organisms. Graft replacement was the most common surgical procedure in patients with GNRB after CVS (44.9%). Prophylaxis antibiotics were ampicillin/sulbactam (76.9%), and vancomycin (12.8%). The crude 90-day mortality rate was 21.8%, and the mean Acute Physiology and Chronic Health Evaluation II score was 15.6 (range, 3-39). In 34.6% of patients, the same GNR species were isolated from other samples within 30 days of GNRB occurrence. Multivariate analysis indicated that P. aeruginosa bacteremia [odds ratio (OR), 175; confidence interval (CI), 2.40-1270; p = 0.0182], Acute Physiology and Chronic Health Evaluation II scores of ≥ 25 (OR 76.2; CI 1.04-5580; p = 0.0479), and vancomycin for prophylaxis (OR 45.4; CI 1.02-202; p = 0.0488) were significant independent prognostic factors associated with death due to GNRB after CVS., Conclusion: Graft replacement was the most common surgical procedure in patients with GNRB after CVS. Empirical antibiotics covering Gram-negative rods including P. aeruginosa should be considered if bacteremia is suspected in unstable patients after CVS., (Copyright © 2015. Published by Elsevier B.V.)

We describe a sensitive plant monitoring system by the detection of the bioelectric potentials in plants with boron-doped diamond (BDD) electrodes. For sensor electrodes, we used commercially available BDD, Ag, and Pt plate electrodes. We tested this approach on a hybrid species in the genus Opuntia (potted) and three different trees (ground-planted) at different places in Japan. For the Opuntia, we artificially induced bioelectric potential changes by the surface potential using the fingers. We detected substantial changes in bioelectric potentials through all electrodes during finger touches on the surface of potted Opuntia hybrid plants, although the BDD electrodes were several times more sensitive to bioelectric potential change compared to the other electrodes. Similarly for ground-planted trees, we found that both BDD and Pt electrodes detected bioelectric potential change induced by changing environmental factors (temperature and humidity) for months without replacing/removing/changing electrodes, BDD electrodes were 5-10 times more sensitive in this detection than Pt electrodes. Given these results, we conclude that BDD electrodes on live plant tissue were able to consistently detect bioelectrical potential changes in plants.

Background: Clostridium difficile infection (CDI) is a highly prevalent hospital-associated infection. Although most patients respond well to discontinuation of antibiotics, 20%-30% of patients relapse. To initiate early therapeutic measures, the risk factors for recurrent CDI must be identified, although very few Japanese studies have used standard surveillance definitions to identify these risk factors., Methods: We retrospectively reviewed the medical records of patients with health care facility-onset CDI between August 2011 and September 2013. Patients with diarrhea who were positive for Clostridium difficile (via an enzyme immunoassay) were defined as having CDI. Clinical data (eg, demographics, comorbidities, medication, laboratory results, and clinical outcomes) were evaluated, and multivariate analysis was used to identify risk factors that were associated with recurrent CDI., Results: Seventy-six health care facility-onset CDI cases were identified, with an incidence rate of 0.8 cases per 10,000 patient-days. Fourteen cases (18.4%) were recurrent, with 13 patients having experienced a single recurrent episode and one patient having experienced three recurrent episodes. The 30-day and 90-day mortality rates were 7.9% and 14.5%, respectively. Multivariate analysis revealed that recurrent patients were more likely to have underlying malignant disease (odds ratio: 7.98; 95% confidence interval: 1.22-52.2; P=0.03) and a history of intensive care unit hospitalization (odds ratio: 49.9; 95% confidence interval: 1.01-2,470; P=0.049)., Conclusion: Intensive care unit hospitalization and malignancy are risk factors for recurrent CDI. Patients with these factors should be carefully monitored for recurrence and provided with appropriate antimicrobial stewardship.

Dynasore, a specific dynamin GTPase inhibitor, suppresses lamellipodia formation and cancer cell invasion by destabilizing actin filaments. In search for novel dynamin inhibitors that suppress actin dynamics more efficiently, dynasore analogues were screened. N'-[4-(dipropylamino)benzylidene]-2-hydroxybenzohydrazide (DBHA) markedly reduced in vitro actin polymerization, and dose-dependently inhibited phosphatidylserine-stimulated dynamin GTPase activity. DBHA significantly suppressed both the recruitment of dynamin 2 to the leading edge in U2OS cells and ruffle formation in H1299 cells. Furthermore, DBHA suppressed both the migration and invasion of H1299 cells by approximately 70%. Furthermore, intratumoral DBHA delivery significantly repressed tumor growth. DBHA was much less cytotoxic than dynasore. These results strongly suggest that DBHA inhibits dynamin-dependent actin polymerization by altering the interactions between dynamin and lipid membranes. DBHA and its derivative may be potential candidates for potent anti-cancer drugs., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Up to date, no worldwide standard in vitro method has been established for the determination of the sun protection factor (SPF), since there are many problems in terms of its repeatability and reliability. Here, we have studied the problems on the in vitro SPF measurements brought about by the phenomenon called viscous fingering. A spatially periodic stripe pattern is usually formed spontaneously when a viscous fluid is applied onto a solid substrate. For the in vitro SPF measurements, the recommended amount of sunscreen is applied onto a substrate, and the intensity of the transmitted UV light through the sunscreen layer is evaluated. Our theoretical analysis indicated that the nonuniformity of the thickness of the sunscreen layer varied the net UV absorbance. Pseudo-sunscreen composites having no phase separation structures were prepared and applied on a quartz plate for the measurements of the UV absorbance. Two types of applicators, a block applicator and a 4-sided applicator were used. The flat surface was always obtained when the 4-sided applicator was used, while the spatially periodic stripe pattern was always generated spontaneously when the block applicator was used. The net UV absorbance of the layer on which the stripe pattern was formed was found to be lower than that of the flat layer having the same average thickness. Theoretical simulations quantitatively reproduced the variation of the net UV absorbance led by the change of the geometry of the layer. The results of this study propose the definite necessity of strict regulations on the coating method of sunscreens for the establishment of the in vitro SPF test method., (© 2014 S. Karger AG, Basel.)

Dynamin GTPase, a key molecule in endocytosis, mechanically severs the invaginated membrane upon GTP hydrolysis. Dynamin functions also in regulating actin cytoskeleton, but the mechanisms are yet to be defined. Here we show that dynamin 1, a neuronal isoform of dynamin, and cortactin form ring complexes, which twine around F-actin bundles and stabilize them. By negative-staining EM, dynamin 1-cortactin complexes appeared as "open" or "closed" rings depending on guanine nucleotide conditions. By pyrene actin assembly assay, dynamin 1 stimulated actin assembly in mouse brain cytosol. In vitro incubation of F-actin with both dynamin 1 and cortactin led to the formation of long and thick actin bundles, on which dynamin 1 and cortactin were periodically colocalized in puncta. A depolymerization assay revealed that dynamin 1 and cortactin increased the stability of actin bundles, most prominently in the presence of GTP. In rat cortical neurons and human neuroblastoma cell line, SH-SY5Y, both dynamin 1 and cortactin localized on actin filaments and the bundles at growth cone filopodia as revealed by immunoelectron microscopy. In SH-SY5Y cell, acute inhibition of dynamin 1 by application of dynamin inhibitor led to growth cone collapse. Cortactin knockdown also reduced growth cone filopodia. Together, our results strongly suggest that dynamin 1 and cortactin ring complex mechanically stabilizes F-actin bundles in growth cone filopodia. Thus, the GTPase-dependent mechanochemical enzyme property of dynamin is commonly used both in endocytosis and regulation of F-actin bundles by a dynamin 1-cortactin complex.

The present study was conducted to investigate the effect of estradiol treatment and/or ovariectomy (OVX) on non-neoplastic lesions in the pancreatic islets of Sprague-Dawley rats. Males were divided into non-treatment (naïve) and beta-estradiol 3-benzoate (EB) treatment groups and females into naïve, sham-operation, OVX, and OVX plus EB treatment groups. EB was subcutaneously administered once a week from seven to twenty-six weeks of age. The animals were euthanized at twelve, eighteen, and twenty-six weeks of age, and the serum estradiol concentrations were measured in conjunction with the pancreatic islet histopathology. The histological stages of pancreatic findings were classified into three groups, hemorrhagic, fibrotic, and inflammatory lesions, and the incidence of each type of lesion was enumerated. In males, both the total and individual incidence of pancreatic lesions increased age dependently in the naïve group. EB treatment significantly decreased the total incidence at twenty-six weeks. This alteration consisted of fibrotic and inflammatory lesions, but not hemorrhagic lesions. Additionally, the incidence of hemorrhagic lesions was at the same level between male naïve and male EB groups at twelve weeks, despite a markedly higher concentration of serum estradiol in the EB group. In females, a similar tendency was seen, and the total incidence was generally low in the naïve group, whereas it was increased by OVX. OVX plus EB treatment tended to decrease the incidence accompanied by a marked increase in estradiol concentrations. In conclusion, estrogen was shown to inhibit the development of pancreatic islet lesions toward inflammation and fibrosis but did not inhibit the occurrence of hemorrhagic lesions.

A gold-catalyzed alkylation of alcohols and aromatic compounds is described. The reaction of ortho-alkynylbenzoic acid alkyl esters with alcohols or aromatic compounds occurs in the presence of catalytic amounts of Ph3PAuCl and AgOTf under mild conditions to produce corresponding ethers or Friedel-Crafts alkylation products in good to high yields. The reaction likely proceeds through the gold-induced in situ construction of leaving groups and subsequent nucleophilic attack of alcohols or aromatic compounds.

We analyzed the proteome of a crenararchaeon, Aeropyrum pernix K1, by using the following four methods: (i) two-dimensional PAGE followed by MALDI-TOF MS, (ii) one-dimensional SDS-PAGE in combination with two-dimensional LC-MS/MS, (iii) multidimensional LC-MS/MS, and (iv) two-dimensional PAGE followed by amino-terminal amino acid sequencing. These methods were found to be complementary to each other, and biases in the data obtained in one method could largely be compensated by the data obtained in the other methods. Consequently a total of 704 proteins were successfully identified, 134 of which were unique to A. pernix K1, and 19 were not described previously in the genomic annotation. We found that the original annotation of the genomic data of this archaeon was not adequate in particular with respect to proteins of 10-20 kDa in size, many of which were described as hypothetical. Furthermore the amino-terminal amino acid sequence analysis indicated that surprisingly the translation of 52% of their genes starts with TTG in contrast to ATG (28%) and GTG (20%). Thus, A. pernix K1 is the first example of an organism in which TTG is the most predominant translational initiation codon.

Rhodococcus erythropolis strain PR4 has been isolated as an alkane-degrading bacterium. The strain harbours one linear plasmid, pREL1 (271 577 bp) and two circular plasmids, pREC1 (104 014 bp) and pREC2 (3637 bp), all with some sequence similarities to other Rhodococcus plasmids. For pREL1, pREC1 and pREC2, 298, 102 and 3 open reading frames, respectively, were predicted. Linear plasmid pREL1 has several regions homologous to plasmid pBD2 found in R. erythropolis BD2. Sequence analysis of pREL1 and pBD2 identified common metal-resistance genes on both, but pREL1 also encodes alkane-degradation genes not found on pBD2, with enzyme constituents some of which are quite different from those of other organisms. The alkane hydroxylase consisted of a cytochrome P450 monooxygenase, a 2Fe-2S ferredoxin, and a ferredoxin reductase. The ferredoxin reductase amino acid sequence resembles the AlkT (rubredoxin reductase) sequence. A zinc-containing alcohol dehydrogenase further oxydizes alkanols, alkane oxidation products catalysed by alkane hydroxylase. Of the circular plasmids, the pREC1 sequence is partially similar to the sequence of pREAT701, the virulence plasmid found in Rhodococcus equi. pREC1 has no pREAT701 virulence genes and encodes genes for beta-oxidation of fatty acids. Thus, joint actions of enzymes encoded by pREL1 and pREC1 may enable efficient mineralization of alkanes.

We report 23 years of observation of a patient with pycnodysostosis associated with progressive spondylolysis. There have been very few papers describing the development of spondylolysis associated with pycnodysostosis as confirmed by serial X-ray examinations. The diagnosis was made by the patient's typical bird-like face, plain radiological findings, and repeated tibial fractures. At the age of 8 years, spondylolysis of L4 was observed. The spondylolysis had increased in number to 4 by the age of 24 years, that is, L2 through L5.

A 46-year-old Japanese woman underwent a right lower lobectomy through a posterolateral incision made in the fifth intercostal space under general and epidural anesthesia on January 23, 1995. During the procedure, oxidized regenerated cellulose (Surgicel) was used to prevent postoperative rebleeding from the dorsal branch of the fifth intercostal artery. The following day it became evident that complete paraplegia had developed below the Th5 level, the cause of which was revealed by an emergency laminectomy, performed within 20 h after the thoracotomy, to be the Surgical treatment. By 50 days after the operation the patient had begun to show improvement, and was able to move her lower extremities against gravity. Her condition is continuing to improve.