A deazariboflavin chromophore kinetically stabilizes reduced FAD state in a bifunctional cryptochrome

An animal-like cryptochrome derived from Chlamydomonas reinhardtii (CraCRY) is a bifunctional flavoenzyme harboring flavin adenine dinucleotide (FAD) as a photoreceptive/catalytic center and functions both in the regulation of gene transcription and the repair of UV-induced DNA lesions in a light-dependent manner, using different FAD redox states. To address how CraCRY stabilizes the physiologically relevant redox state of FAD, we investigated the thermodynamic and kinetic stability of the two-electron reduced anionic FAD state (FADH−) in CraCRY and related (6–4) photolyases. The thermodynamic stability of FADH− remained almost the same compared to that of all tested proteins. However, the kinetic stability of FADH− varied remarkably depending on the local structure of the secondary pocket, where an auxiliary chromophore, 8-hydroxy-7,8-didemethyl-5-deazariboflavin (8-HDF), can be accommodated. The observed effect of 8-HDF uptake on the enhancement of the kinetic stability of FADH− suggests an essential role of 8-HDF in the bifunctionality of CraCRY.Hosokawa Y., Morita H., Nakamura M., et al. A deazariboflavin chromophore kinetically stabilizes reduced FAD state in a bifunctional cryptochrome. Scientific Reports 13, 16682 (2023); https://doi.org/10.1038/s41598-023-43930-0

Cystatin C in children with malignancies

Several factors besides renal function influence serum cystatin C (CysC) levels. The present study evaluates the value of serum CysC and the equation for CysC based estimated glomerular filtration rate (CysC-eGFR) for Japanese children with malignancies.We collected information at 36 time points from 13 patients aged ≤ 17 years with malignancies. We assessed tumor activity, cell recovery phase after chemotherapy, neutropenia phase, inflammation response and medication with granulocyte-colony stimulating factor, steroid, and levothyroxine as risk factors associated with serum CysC levels. Although no 24-h creatinine clearance (CCr) data collected at 36 time points indicated renal dysfunction, serum CysC levels were above and below the reference values at four and five time points, respectively. The frequency of elevated serum CysC levels was higher in patients without therapy or with stable or progressive disease than among those with a complete or partial response (p = 0.0046). The correlation coefficient between CCr and CysC-eGFR was 0.355 (p = 0.054), but this improved to 0.663 (p = 0.0010) when restricted to patients with a complete or partial response. Levels of serum CysC might become elevated regardless of renal function, and CysC-eGFR might become unpredictable during the active phase of tumors

Broadband dielectric spectroscopy of glucose aqueous solution: Analysis of the hydration state and the hydrogen bond network.

Recent studies of saccharides' peculiar anti-freezing and anti-dehydration properties point to a close association with their strong hydration capability and destructuring effect on the hydrogen bond (HB) network of bulk water. The underlying mechanisms are, however, not well understood. In this respect, examination of the complex dielectric constants of saccharide aqueous solutions, especially over a broadband frequency region, should provide interesting insights into these properties, since the dielectric responses reflect corresponding dynamics over the time scales measured. In order to do this, the complex dielectric constants of glucose solutions between 0.5 GHz and 12 THz (from the microwave to the far-infrared region) were measured. We then performed analysis procedures on this broadband spectrum by decomposing it into four Debye and two Lorentz functions, with particular attention being paid to the β relaxation (glucose tumbling), δ relaxation (rotational polarization of the hydrated water), slow relaxation (reorientation of the HB network water), fast relaxation (rotation of the non-HB water), and intermolecular stretching vibration (hindered translation of water). On the basis of this analysis, we revealed that the hydrated water surrounding the glucose molecules exhibits a mono-modal relaxational dispersion with 2-3 times slower relaxation times than unperturbed bulk water and with a hydration number of around 20. Furthermore, other species of water with distorted tetrahedral HB water structures, as well as increases in the relative proportion of non-HB water molecules which have a faster relaxation time and are not a part of the surrounding bulk water HB network, was found in the vicinity of the glucose molecules. These clearly point to the HB destructuring effect of saccharide solutes in aqueous solution. The results, as a whole, provide a detailed picture of glucose-water and water-water interactions in the vicinity of the glucose molecules at various time scales from sub-picosecond to hundreds of picoseconds

Experimental Assessment of Restoration of the Bronchial Artery in Sleeve Lobectomy Combined with Pulmonary Angioplasty

The restoration of the bronchial artery after bronchoplasty combined with pulmonary angioplasty was studied in relation to the degree of stenosis of the pulmonary artery by means of the microangiographic technique. Thirty-three dogs were used in this study. They were divided into Group A (15 dogs) with sleeve lobectomy alone, Group B (10 dogs) with sleeve lobectomy and a 50% stenosis of the left main pulmonary artery and Group C (8 dogs) with sleeve lobectomy and a 75% stenosis of the left main pulmonary artery. The restoration of the bronchial artery was assessed by microangiography. 1) The restoration of th bronchial artery after bronchoplasty was completed on day 14. The trend toward a ready regeneration of the bronchial artery becomes apparent if some degree of stenosis of the pulmonary artery exists after a duration of seven days. 2) The interrupted bronchial artery starts to restore at the adventitial face and extends throughout the whole wall of the bronchus after the 7th day. 3) From these results, an operative procedure of sleeve lobectomy with pulmonary angioplasty is applicable even though stenosis of the pulmonary artery remains to some degree

Risk of bleeding and repeated bleeding events in prasugrel-treated patients: a review of data from the Japanese PRASFIT studies

Prasugrel is a third-generation thienopyridine that achieves potent platelet inhibition with less pharmacological variability than other thienopyridines. However, clinical experience suggests that prasugrel may be associated with a higher risk of de novo and recurrent bleeding events compared with clopidogrel in Japanese patients undergoing percutaneous coronary intervention (PCI). In this review, we evaluate the risk of bleeding in Japanese patients treated with prasugrel at the doses (loading/maintenance doses: 20/3.75 mg) adjusted for Japanese patients, evaluate the risk factors for bleeding in Japanese patients, and examine whether patients with a bleeding event are at increased risk of recurrent bleeding. This review covers published data and new analyses of the PRASFIT (PRASugrel compared with clopidogrel For Japanese patIenTs) trials of patients undergoing PCI for acute coronary syndrome or elective reasons. The bleeding risk with prasugrel was similar to that observed with the standard dose of clopidogrel (300/75 mg), including when bleeding events were re-classified using the Bleeding Academic Research Consortium criteria. The pharmacodynamics of prasugrel was not associated with the risk of bleeding events. The main risk factors for bleeding events were female sex, low body weight, advanced age, and presence of diabetes mellitus. Use of a radial puncture site was associated with a lower risk of bleeding during PCI than a femoral puncture site. Finally, the frequency and severity of recurrent bleeding events during continued treatment were similar between prasugrel and clopidogrel. In summary, this review provides important insights into the risk and types of bleeding events in prasugrel-treated patients. Trial registration numbers: JapicCTI-101339 and JapicCTI-111550

Dispersion-Flocculation Behavior of Fine Lead Particles in an Organic Solvent

Methyl ethyl ketone (MEK) is a good solvent for polyvinyl chloride (PVC) and it has been proposed for use in PVC recycling. In the recycling process, fine particles of 3PbOÁPbSO 4 ÁH 2 O, used as a thermal stabilizer in PVC products, are dispersed and not dissolved in the solvent. To establish methods for removing of 3PbOÁPbSO 4 ÁH 2 O particles from the solvent, factors affecting the dispersion-flocculation behavior of the particles in MEK were investigated. The zeta potential and particle distribution of 3PbOÁPbSO 4 ÁH 2 O particles in MEK solutions containing known amounts of H 2 O were measured. Above 5 vol%H 2 O in MEK solutions, the zeta potential of 3PbOÁPbSO 4 ÁH 2 O particles approached zero and the flocculation of particles was achieved. In addition, it was found that Pb 2þ and Cl À affect the zeta potential of the particles. These results indicate that the dispersion-flocculation behavior of lead particles can be influenced by the concentration of H 2 O, Pb 2þ , and Cl À in MEK

Anti-glycopeptide mAb LpMab-21 against Podoplanin

Human podoplanin (hPDPN), which binds to C‐type lectin‐like receptor‐2 (CLEC‐2), is involved in platelet aggregation and cancer metastasis. The expression of hPDPN in cancer cells or cancer‐associated fibroblasts indicates poor prognosis. Human lymphatic endothelial cells, lung‐type I alveolar cells, and renal glomerular epithelial cells express hPDPN. Although numerous monoclonal antibodies (mA bs) against hPDPN are available, they recognize peptide epitopes of hPDPN. Here, we generated a novel anti‐hPDPN mA b, LpMab‐21. To characterize the hPDPN epitope recognized by the LpMab‐21, we established glycan‐deficient CHO‐S and HEK‐293T cell lines, using the CRISPR/Cas9 or TALEN. Flow cytometric analysis revealed that the minimum hPDPN epitope, in which sialic acid is linked to Thr76, recognized by LpMab‐21 is Thr76–Arg79. LpMab‐21 detected hPDPN expression in glioblastoma, oral squamous carcinoma, and seminoma cells as well as in normal lymphatic endothelial cells. However, LpMab‐21 did not react with renal glomerular epithelial cells or lung type I alveolar cells, indicating that sialylation of hPDPN Thr76 is cell‐type‐specific. LpMab‐21 combined with other anti‐hPDPN antibodies that recognize different epitopes may therefore be useful for determining the physiological function of sialylated hPDPN

Effect of CYP3A5*3 genetic variant on the metabolism of direct-acting antivirals in vitro : a different effect on asunaprevir versus daclatasvir and beclabuvir

Direct-acting antivirals, asunaprevir (ASV), daclatasvir (DCV), and beclabuvir (BCV) are known to be mainly metabolized by CYP3A enzymes; however, the differences in the detailed metabolic activities of CYP3A4 and CYP3A5 on these drugs are not well clarified. The aim of the present study was to elucidate the relative contributions of CYP3A4 and CYP3A5 to the metabolism of ASV, DCV, and BCV, as well as the effect of CYP3A5*3 genetic variant in vitro. The amount of each drug and their major metabolites were determined using LC-MS/MS. Recombinant CYP3As and CYP3A5*3-genotyped human liver microsomes (CYP3A5 expressers or non-expressers) were used for the determination of their metabolic activities. The contribution of CYP3A5 to ASV metabolism was considerable compared to that of CYP3A4. Consistently, ASV metabolic activity in CYP3A5 expressers was higher than those in CYP3A5 non-expresser. Moreover, CYP3A5 expression level was significantly correlated with ASV metabolism. In contrast, these observations were not found in DCV and BCV metabolism. To our knowledge, this is the first study to directly demonstrate the effect of CYP3A5*3 genetic variants on the metabolism of ASV. The findings of the present study may provide basic information on ASV, DCV, and BCV metabolisms

Results of Surgical Treatment for Small Cell Lung Cancers

We analyzed the data for patients with small cell lung cancer, especially as regards comparison of the results of surgical treatment before and after the introduction of multimodal chemotherapy treatments. Sixty cases with small cell lung cancer were admitted in our department between January, 1955 and December, 1993. Among them, 38 cases underwent pulmonary resection. To evaluate the efficacy of the multimodal treatment including cisplatin with surgical therapy, patients were devided whether pulmonary resection was performed before 1983 (Group A) or after 1984 (Group B). There were no differences in sex and stage between two groups, but ages and operative proceduress were significantly different (p <0.05). Ages were older in Group B (65.5 years) than in Group A (58.1 years) and operations were lesser in Group B (lobectomy or segmentectomy 87.0 %) than in Group A (pneumonectomy or bilobectomy 53.5 %). The operative mortality rates were 13.3 % (2/15) in Group A and 4.3 % (1/23) n Group B. Survival rate at 3 years of Group A was only 6.7 % and no patients survived more than 4 years. While, survival rates of Group B at 3 and 5 years were 24.2 %. The 5 years survival rate of patients with Stage I and II of Group B was 46.9 % and that of Stage III and IV was 8.5%. It is concluded that surgical resection of limited small cell lung cancer (Stage I and II) with intensive chemotherapy is an efficient therapeutic approach