Evaluation of cleaning methods for change-over after the processing of cell products to avoid cross-contamination risk.

Introduction: Cell-processing facilities face the risk of environmental bacteria contaminating biosafety cabinets during processing, and manual handling of autologous cell products can result in contamination. We propose a risk- and evidence-based cleaning method for cross-contamination, emphasizing proteins and DNA. Methods: The transition and residual risks of the culture medium were assessed by measuring both wet and dried media using fluorescence intensity. Residual proteins and DNA in dried culture medium containing HT-1080 cells were analyzed following ultraviolet (UV) irradiation, wiping, and disinfectant treatment. Results: Wet conditions showed a higher transition to distilled water (DW), whereas dry conditions led to higher residual amounts on SUS304 plates. Various cleaning methods for residual culture medium were examined, including benzalkonium chloride with a corrosion inhibitor (BKC + I) and DW wiping, which demonstrated significantly lower residual protein and DNA compared to other methods. Furthermore, these cleaning methods were tested for residual medium containing cells, with BKC + I and DW wiping resulting in an undetectable number of cells. However, in some instances, proteins and DNA remained. Conclusions: The study compared cleaning methods for proteins and DNA in cell products, revealing their advantages and disadvantages. Peracetic acid (PAA) proved effective for nucleic acids but not proteins, while UV irradiation was ineffective against both proteins and DNA. Wiping emerged as the most effective method, even though traceability remained challenging. However, wiping with ETH was not effective as it caused protein immobilization. Understanding the characteristics of these cleaning methods is crucial for developing effective contamination control strategies.

Synthesis and evaluation of fluorescent resins with europium-ß-diketonate complex for orthodontic use.

No effective technique exists for removing adhesive remnants following bracket debonding. We propose that fluorescence imaging using europium ions (Eu3+) offers an effective solution for minimizing iatrogenic enamel damage. This study aimed to assess the impact of different mixing ratios of monomer mixtures on the photoluminescence and flexural properties of a newly developed fluorescent adhesive. Four monomer blends with varying urethane dimethacrylate (UDMA) to triethylene glycol dimethacrylate (TEGDMA) ratios were prepared and polymerized. The blends contained 0.1 wt% of tris(1,3-diphenyl-1,3-propanedionato)(1,10-phenanthroline) Eu(III), [Eu(DBM)3Phen], as the phosphor. Optical measurements and flexural tests were conducted for each resin specimen. The emission spectra exhibited narrow bands corresponding to the 4f-4f transitions of the Eu3+ ions. The photoluminescence properties remained unaffected by the mixing ratio, whereas the mechanical properties tended to improve with higher UDMA content. We conclude that the Eu(DBM)3Phen-containing resin shows promise as a fluorescent orthodontic adhesive that contributes to preserving enamel health.

Evaluation of the meropenem dosage and administration schedule in patients with bacteremia initial therapy.

BACKGROUND: The standard meropenem (MEPM) regimen allowed by insurance in Japan is 0.5 g two or three times a day. Differences in dosages and administration schedules in Japan were evaluated. METHODS: Patients with bacteremia for whom MEPM was used as the initial treatment at our institution between 2016 and 2021 were included. We retrospectively investigated patients classified into two groups: those treated according to severe infections (high-dose groupand others (low-dose group). After propensity score matching, we compared the probability of achieving free drug blood levels above the minimum inhibitory concentration (MIC) in 24 h (%fT > MIC) and outcomes. RESULTS: The probability of 100% fT > MIC was significantly higher in the high-dose group (96.4% vs 74.5%, odds ratio [OR] = 0.3, 95% confidence interval [CI] = 0.2-0.4, P = < 0.001). Regarding outcomes, the 30-day mortality rate was significantly lower in the high-dose group (1.4% vs. 11.4%, OR = 8.0, 95% CI = 1.5-43.7, P = 0.019). CONCLUSIONS: To improve outcomes in patients with bacteremia treated with MEPM, support for appropriate antimicrobial use is necessary for compliance with the dosage and administration schedule according to severe infections in initial treatment.

Tungsten(VI) Complex of N-Fused Porphyrin Absorbing Near-Infrared Light beyond 1000 nm.

Incorporating tungsten into the N3 core of a N-fused porphyrin (NFP; 1) affords high-valent tungsten(VI)-NFP complexes, WClO2 -1 and 21-chlorinated WClO2 -3. The X-ray structure of WClO2 -1 reveals a distorted octahedral geometry with sitting atop metal coordination. The absorption spectrum of WClO2 -1 displays bathochromically shifted Q-like bands beyond 1000 nm, indicating an inherently narrow HOMO-LUMO energy gap. DFT calculations show that the degeneracy of the LUMO and LUMO+1 pair of 1 is significantly resolved by the tungsten(VI) coordination. Conclusively, magnetic circular dichroism (MCD) spectroscopy and cyclic voltammetry provide a rationale for the narrow HOMO-LUMO energy gap in the "16-electron" d0 tungsten(VI)-NFP complexes.

Ruthenium N-Confused Porphyrins: Selective Reactivity for Ambident 2-Heteroatom-Substituted Pyridines Serving as Axial Ligands.

Three types of ruthenium(II) N-confused porphyrin (NCP) complexes bearing an axial 2-thiopyridine, 2-pyridone, and 2-iminopyridine moiety at the inner carbon atom, respectively, were synthesized. The unique reactivity of the 2-substituted pyridine derivatives (2-X-pyridine; X=SH, OH, NH2 ) toward the inner carbon atom of the NCP allows the formation of two types of coordinated products (i. e., pyridine donor versus 2-heteroatom donors), as inferred from single-crystal X-ray structures. The selective reactivity was investigated by using density functional theory (DFT) calculations. Finally, the catalytic activity of these ruthenium complexes was demonstrated through the styrene oxidation reactions. As a result, the ruthenium(II) NCP complex bearing a 2-thiopyridine moiety, together with aqueous H2 O2 as an oxidant showed the highest selectivity for benzaldehyde (benzaldehyde/styrene oxide=20 : 1).

Two Discrete RuCp* (Cp*=Pentamethylcyclopentadienyl) Binding Modes of N-Confused Porphyrins: Peripheral π Complex and Sitting Atop Ruthenocenophane Complex by Skeletal Transformation.

Complexation of a RuCp* cation with N-confused tetraarylporphyrins (NCPs) forms directly bound ruthenium(II) pentamethylcyclopentadienyl (Cp*) π-complex on a specific meso-aryl group (e.g., phenyl) neighboring peripheral imino nitrogen of NCPs in high yields. In contrast, in the case of NCPs bearing bulky meso-substituents (e.g., 3,5-di-tert-butylphenyl), new ruthenocenophane-like complex embedded on an N-confused calix[4]phyrin was formed through multiple C-H bond activation of methyl groups of Cp* ligand. The mechanistic insight into the formation of the ruthenocenophane was derived from DFT calculations.

Characterization of a recombinant Bacteroides fragilis sialidase expressed in Escherichia coli.

The human gut commensal Bacteroides fragilis produces sialidases that remove a terminal sialic acid from host-derived polysaccharides. Sialidase is considered to be involved in B. fragilis infection pathology. A native B. fragilis sialidase has been purified and characterized, and was shown to be post-translationally modified by glycosylation. However, the biochemical properties of recombinant B. fragilis sialidase expressed in a heterologous host remain uncharacterized. In this study, we examined the enzymatic properties of the 60-kDa sialidase NanH1 of B. fragilis YCH46, which was prepared as a recombinant protein (rNanH1) in Escherichia coli. In E. coli rNanH1 was expressed as inclusion bodies, which were separated from soluble proteins to allow solubilization of insoluble rNanH1 in a buffer containing 8 M urea and renaturation in refolding buffer containing 100 mM CaCl2 and 50 mM L-arginine. The specific activity of renatured rNanH1 measured using 4-methylumberiferyl-α-D-N-acetyl neuraminic acid as a substrate was 6.16 µmol/min/mg. The optimal pH of rNanH1 ranged from 5.0 to 5.5. The specific activity of rNanH1 was enhanced in the presence of calcium ions. rNanH1 preferentially hydrolyzed the sialyl α2,8 linkage and cleaved sialic acids from mucin and serum proteins (e.g., fetuin and transferrin) but not from α1-acid glycoprotein, which is similar to the previously observed biochemical properties for a native sialidase purified from B. fragilis SBT3182. The results and methods described in this study will be useful for preparing and characterizing recombinant proteins for other B. fragilis sialidase isoenzymes.

Evaluation of the effectiveness of intermittent mechanical pressure with short loading duration: new type of intermittent force for orthodontic treatment.

Background: Since it is difficult to precisely control the magnitude of force in orthodontic practice, controlling the duration of force is expected to prevent tissue damage. The use of an actuator as an appliance for intermittent force application is a possible solution for controlling the duration. However, effectiveness of an actuator for controlling the duration is still not clear. Aim: To determine whether a short loading duration of intermittent force has the potential to maintain a sound condition of the capillaries and induce dilation of capillaries, which is a requisite phenomenon for orthodontic tooth movement. Materials and methods: Six-week-old male hamsters with a dorsal skinfold chamber attached to an intermittent loading device were used. In three experimental groups, application of intermittent pressure that involved repetition of loading and unloading was performed. The durations of loading/unloading were 1 second/9 seconds (group T10), 1 second/19 seconds (group T20), and 1 second/29 seconds (group T30). Vessels were examined using a fluorescence microscope and a stereomicroscope for 5 days. Results: Widths of capillaries in groups T10 and T20 increased significantly (P < 0.01). In contrast, widths of capillaries in group T30 showed no significant difference from those in the control group. Only group T10 showed bleeding, obvious destruction of vessels, and a significant increase in the rate of disappearance of vessels. Conclusion: In the epidermis, although conditions are limited, a short loading duration of intermittent force maintains a sound condition of the capillaries at the tissue level and induces dilation of capillaries.

Cooperation between metal and ligand in oxygen atom transport by N-confused porphyrin oxorhenium(V) complexes.

N-Confused porphyrin oxorhenium(V) complexes were prepared and their X-ray structures were elucidated. The oxorhenium(V) complexes can transfer oxygen atom from pyridine N-oxide to triphenylphosphine, in which unique cooperation between metal and ligand was observed.

Regulation of NH-tautomerism in N-confused porphyrin by N-alkylation.

A variety of internally N-alkylated N-confused porphyrins were prepared in a stepwise manner through the protection of the reactive peripheral nitrogen atom. NH-Tautomerism in N-confused porphyrins was found to be regulated by N-alkylation, which enabled us to obtain discrete information on two important NH-tautomers of an N-confused porphyrin.

[Two patients with recurrent colon cancer who underwent surgery following a combination of irinotecan and UFT].

One patient with pulmonary metastasis from colon cancer and one with para-aortic lymph node metastasis were treated with a combination of irinotecan and UFT. Irinotecan (100 mg/m2) was given by 24-hour intravenous infusion on day 1, and UFT (600 mg/day) was given orally on days 3 to 7 and days 10 to 14 of a 2-week course, which was then repeated. In the patient with pulmonary metastasis, the lesions in the lung resolved after 7 courses of chemotherapy. Surgery was performed after 10 courses. The patient with para-aortic lymph node metastasis had a partial response after 4 courses of chemotherapy, and underwent surgery after 6 courses. The only adverse effects were grade 2 myelosuppression and hair loss, none of which were severe enough to require treatment. With this chemotherapy regimen, patients are admitted for two days biweekly for 24-hour intravenous infusion of CPT-11. Thus, most of the treatment can be performed on an outpatient basis. The combination of irinotecan and UFT is expected to be useful for metastatic or recurrent colon cancer.