Comparison of Microorganism Detection, Time to Positivity, and Time-Dependent Shift in Viable Bacterial Count from VersaTREK and BacT/ALERT 3D Blood Culture Systems.

Rapid positive blood culture reporting allows early and appropriate treatment of severe infections to improve patient prognosis. This study evaluated performance of the VersaTREK system with gas pressure detection and tornado stirring method and the conventional BacT/ALERT 3D system. Time to positivity (TTP) of simulated blood cultures without whole blood using 17 ATCC strains was faster with VersaTREK than BacT/ALERT 3D, averaging 6.3 h in aerobic bottles and 12.7 hours in anaerobic bottles. In simulated blood cultures with whole blood using 53 clinical isolates, on average, VersaTREK was faster in aerobic bottles by 6.5 h but slower in anaerobic bottles by 3.8 h. Fifty of 53 simulated blood cultures with whole blood (94%) showed fastest TTP with VersaTREK. TTP of VersaTREK for anaerobic bacteria Bacteroides fragilis and Clostridium perfringens, Helicobacter cinaedi, and Candida glabrata was fast, and viable bacteria numbers in bottles using the Miles and Misra method increased quickly.

Modulating effects of oral administration of Lycii Fructus extracts on UVB-induced skin erythema: A Randomized, placebo-controlled study.

Severe UV exposure induces skin inflammation, causing erythema. Lycii Fructus (Lycium barbarum and Lycium chinense) is a potential antioxidant agent with a high content of polyphenols, including rutin and chlorogenic acid. This study examined the effects of Lycii Fructus extract (LFE) on UVB-induced skin erythema in humans. Healthy volunteers were randomly assigned to one of two groups and received UVB irradiation at 1.5 minimal erythemal dose (MED) on day 0 at three designated sites on their backs, and the skin color was measured until day 7. After an 8-week treatment with LFE (900 mg/day) or placebo, UVB irradiation (l.5 MED) was applied again at different sites on day 63. Skin color was continuously measured in each group until day 69. LFE tablet administration for 8 weeks significantly inhibited UVB-induced erythema formation and increased the MED by 13%. Erythema formation peaked on the first day after UVB irradiation, but gradually dissipated over the next several days. LFE tended to accelerate erythema disappearance. To determine the polyphenol responsible for the protection against UVB-induced skin damage, the effects of LFE-derived polyphenols and their metabolites on UVB-induced cytotoxicity were examined in vitro. The major intestinal metabolite of rutin and LFE significantly attenuated phototoxicity and in human keratinocyte HaCaT cells. Quercetin enhanced intracellular glutathione levels in HaCaT cells, even though LFE did not increase it. Together, the results showed that LFE inhibited erythema formation and accelerated erythema dissipation, possibly through its direct antioxidative action.

3,5,7,3',4'-Pentamethoxyflavone Enhances the Barrier Function through Transcriptional Regulation of the Tight Junction in Human Intestinal Caco-2 Cells.

The intestinal tight junction (TJ) barrier plays a pivotal role in the regulation of intestinal homeostasis. This study investigated the effects of 3,5,7,3',4'-pentamethoxyflavone (PMF), a major polymethoxyflavone found in black ginger, on TJ barrier regulation using intestinal Caco-2 cells. PMF treatment enhanced the TJ barrier integrity in Caco-2 cells, indicated by increased transepithelial electrical resistance (control, 1261 ± 36 Ω·cm2; 100 µM PMF, 1383 ± 55 Ω·cm2 at 48 h, p < 0.05) and decreased permeability to fluorescein-conjugated dextran (control, 24.2 ± 1.8 pmol/(cm2 × h); 100 µM PMF, 18.6 ± 1.0 pmol/(cm2 × h), p < 0.05). Immunoblot analysis revealed that PMF increased the cytoskeletal association and cellular expression of the TJ proteins, zonula occludens-1, claudin-3, and claudin-4 (e.g., occludin; control, 1.00 ± 0.2; 100 µM PMF, 3.69 ± 0.86 at 48 h, p < 0.05). Quantitative reverse transcriptase-polymerase chain reaction analysis and a luciferase promoter assay showed that PMF enhanced the transcription of occludin, claudin-3, and claudin-4. The promoter assay with site-directed mutagenesis indicated that PMF-induced occludin and claudin-3 transcription was mediated by transcription factors, KLF5 and EGR1, respectively, while PMF activated claudin-4 transcription through GATA1 and AP1. Taken together, the transcriptional regulation of TJ proteins is involved in PMF-mediated promotion of the intestinal barrier in vitro.

Development of a real-time neutron beam detector for boron neutron capture therapy using a thin silicon sensor.

We have developed a new real-time neutron detector, which is able to measure a direct neutron beam of boron neutron capture therapy. The detector consists of both a 40-µm-thick pn diode and around 0.09-µm-thick LiF neutron converter. Experimental results indicate that this neutron detector can measure neutron flux up to 1 × 109 (cm-2 s-1), separately from gamma rays around 500 mGy/h. The measured depth distribution of neutron flux in an acrylic block is in agreement with the activation results of gold.

A mutation of the ß-domain in POU1F1 causes pituitary deficiency due to dominant PIT-1ß expression.

BACKGROUND: POU1F1 encodes both PIT-1α, which plays pivotal roles in pituitary development and GH, PRL and TSHB expression, and the alternatively spliced isoform PIT-1ß, which contains an insertion of 26-amino acids (ß-domain) in the transactivation domain of PIT-1α due to the use of an alternative splice acceptor at the end of the first intron. PIT-1ß is expressed at much lower levels than PIT-1α and represses endogenous PIT-1α transcriptional activity. Although POU1F1 mutations lead to combined pituitary hormone deficiency (CPHD), no patients with ß-domain mutations have been reported. RESULTS: Here, we report that a three-generation family exhibited different degrees of CPHD, including growth hormone deficiency with intrafamilial variability of prolactin/TSH insufficiency and unexpected prolactinoma occurrence. The CPHD was due to a novel POU1F1 heterozygous variant (c.143-69T>G) in intron 1 of PIT-1α (RefSeq number NM_000306) or as c.152T>G (p.Ile51Ser) in exon 2 of PIT-1ß (NM_001122757). Gene splicing experiments showed that this mutation yielded the PIT-1ß transcript without other transcripts. The lymphocyte PIT-1ß mRNA expression was significantly higher in the patients with the heterozygous mutation than a control. A luciferase reporter assay revealed that the PIT-1ß-Ile51Ser mutant repressed PIT-1α and abolished transactivation capacity for the rat prolactin promoter in GH3 pituitary cells. CONCLUSIONS: We describe, for the first time, that the PIT-1ß mutation can cause CPHD through a novel genetic mechanism, such as PIT-1ß overexpression, and that POU1F1 mutation might be associated with a prolactinoma. Analysis of new patients and long-term follow-up are needed to clarify the characteristics of PIT-1ß mutations.

Oral Supplementation of Collagen Peptides Improves Skin Hydration by Increasing the Natural Moisturizing Factor Content in the Stratum Corneum: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial.

INTRODUCTION: We aimed to investigate the effect of orally ingested collagen peptides (CPs) on skin condition and elucidate their mechanism of action. METHODS: A randomized, placebo-controlled, double-blind trial was conducted in 99 healthy Japanese women, aged 35-50 years. The subjects were randomized into 3 groups (33 subjects/group) to receive 1 or 5 g of CP or placebo once daily for 12 weeks. Skin water content, transepidermal water loss (TEWL), skin elasticity, and skin thickness were evaluated before treatment and after 4, 8, and 12 weeks of treatment. The level of natural moisturizing factor (NMF) constituents in the stratum corneum (SC) was quantified before treatment and after 12 weeks of treatment. RESULTS: Oral ingestion of CP increased the water content in the SC and epidermis and decreased TEWL. Furthermore, the NMF level in the SC was increased. However, skin elasticity and skin thickness remained unchanged. CONCLUSIONS: The improvement in skin water content following the oral ingestion of CP can be attributed to an increase in the level of NMF in the SC. TRIAL REGISTRATION: UMIN000030375 (retrospectively registered).

Novel strategy of rapid typing of Shiga toxin-producing Escherichia coli using MALDI Biotyper and ClinProTools analysis.

INTRODUCTION: Rapid detection of Shiga toxin-producing Escherichia coli (STEC) is essential. Matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows rapid, accurate, and low-cost microbial identification. We aimed to examine the discrimination ability of MALDI Biotyper (Bruker Daltonics) between STEC and non-STEC. METHODS: In total, 234 strains (79 STEC strains and 155 non-STEC strains) isolated from stool between 2009 and 2014 were measured by MALDI Biotyper and mass spectra of 2000-20,000 m/z were analyzed with ClinProTools (Bruker Daltonics). Eighty-three strains were randomly extracted to produce STEC detection models using 3 algorithms, and 151 strains were used as validation samples to verify STEC detection performance and discrimination performance of Shiga toxins with the STEC detection models. RESULTS: The STEC detection model with Quick Classifier (QC) algorithm was the most sensitive: sensitivity, 84.1% (37/44); specificity, 94.4% (101/107). Discrimination between STEC and non-STEC was excellent, but individual discrimination of Shiga toxins was not possible. CONCLUSION: MALDI Biotyper may be a useful rapid diagnostic method for STEC infection.

[A Case of Fungalemia that Detected the Mucor circinelloides by Versa TREK].

Man in his 80s. In March 20XX, the level of consciousness decreased at the admission facility, and he was transported as an emergency case. He was diagnosed as aspiration pneumonia, septic shock due to cholecystitis, and DIC, and was hospitalized for medical treatment. During the course of hospitalization, aspiration pneumonia continued to improve and worsen, but in January 20XX＋3, a fever of 38.7°C occurred, and Mucor circinelloides was detected in the blood culture collected at this time. In sputum 7 days before the blood culture was submitted, an image of suspicious zygomycosis was confirmed by Gram stain, so the patient was diagnosed with Mucor disease and started administration of amphotericin B. After that, the condition was temporarily stable, but due to recurrence of aspiration pneumonia and renal damage, he died 19 days after the start of amphotericin B administration. It is difficult to detect Mucor spp. in blood culture, however in this case, it was detected by the blood culture device; Versa TREK (Thermo Fisher Scientific K.K. Tokyo, Japan).

Correlation of Strain Classification with IR Biotyper and Molecular Epidemiological Method of Pseudomonas aeruginosa.

INTRODUCTION: From 2018, IR Biotyper (IRBT; Bruker Daltonik GmbH, Germany) based on the Fourier transform infrared spectrophotometer has begun to be introduced as a new strain classification method in the field of clinical microbiological examination. We compared it with molecular epidemiology method to evaluate the usefulness of strain classification by IRBT. METHOD: Homology of strain classification with molecular epidemiology method (Multilocus Sequencing Typing; MLST and PCR-based ORF Typing; POT) for 48 strains of Pseudomonas aeruginosa with different detection times from multiple institutions to evaluate the accuracy of IRBT was compared. RESULTS: IRBT used "KBM" SCD agar medium for preculture and was classified into 12 types when classified by Cut-off value 0.181, 8 types by MLST, and 13 types by POT. In the Adjusted Wallace between IRBT and molecular epidemiology method, MLST was 0.458 (95% CI; 0.295 to 0.620) and POT was 0.330 (95% CI; 0.135 to 0.525), indicating a discrepancy in strain classification. CONCLUSION: No correlation was found between IRBT and the classification results by the molecular epidemiology method. In the molecular epidemiology method, strains are classified by matching only specific gene regions, but IRBT irradiates a sample with an infrared laser and classifies the strains according to the difference in absorption spectrum according to the molecular structure, so the measurement principle is different. When classifying strains by IRBT, it is desirable to grasp the clinical information of the detected strains and to target multiple strains isolated at the same facility at the same time.

Single-Domain and Atomically Flat Surface of κ-Ga2O3 Thin Films on FZ-Grown ε-GaFeO3 Substrates via Step-Flow Growth Mode.

Herein, single-domain κ-Ga2O3 thin films were grown on FZ-grown ε-GaFeO3 substrates via a step-flow growth mode. The ε-GaFeO3 possessing the same crystal structure and similar lattice parameters as those of the orthorhombic κ-Ga2O3 facilitated the growth of κ-Ga2O3 thin films, as observed by the X-ray diffraction (XRD) analysis. Furthermore, the surface morphologies of the κ-Ga2O3 thin films exhibited a step-terrace and atomically flat structure. XRD φ-scan and transmission electron microscopy with selected area electron diffraction revealed that there is no occurrence of in-plane rotational domains in the κ-Ga2O3 thin films on ε-GaFeO3 substrates and that the κ-Ga2O3 thin film comprised a single domain. TEM analysis revealed that there were no clear dislocations in the observation area. Moreover, high-resolution TEM observation showed that the atomic arrangements of the film and the substrate were continuous without the presence of an intermediate layer along the growth direction and were well-aligned in the in-plane direction.

A 34-year-old Japanese patient exhibiting NBAS deficiency with a novel mutation and extended phenotypic variation.

Biallelic neuroblastoma amplified sequence (NBAS) gene mutations have recently been identified to cause a reduction in its protein expression and a broad phenotypic spectrum, from isolated short stature, optic nerve atrophy, and Pelger-Huët anomaly (SOPH) syndrome or infantile liver failure syndrome 2 to a combined, multi-systemic disease including skeletal dysplasia and immunological and neurological abnormalities. Herein, we report a 34-year-old patient with a range of phenotypes for NBAS deficiency due to compound heterozygous variants; one is a SOPH-specific variant, p.Arg1914His, and the other is a novel splice site variant, c.6433-2A>G. The patient experienced recurrent acute liver failure until early childhood. Hypogammaglobulinemia, a decrease in natural killer cells, and optic nerve atrophy were evident from infancy to childhood. In adulthood, the patient exhibited novel phenotypic features such as hepatic cirrhosis complicated by portal hypertension and autoimmune hemolytic anemia. The patient also suffered from childhood-onset insulin-requiring diabetes with progressive beta cell dysfunction. The patient had severe short stature and exhibited dysmorphic features compatible with SOPH, intellectual disability, and epilepsy. NBAS protein expression in the patient's fibroblasts was severely low. RNA expression analysis for the c.6433-2A>G variant showed that this variant activated two cryptic splice sites in intron 49 and exon 50, for which the predicted consequences at the protein level were an in-frame deletion/insertion, p.(Ile2199_Asn2202delins16), and a premature termination codon, p.(Ile2199Tyrfs*17), respectively. These findings indicate that NBAS deficiency is a multi-systemic progressive disease. The results of this study extend the spectrum of clinical and genetic findings related to NBAS deficiency.

Accurate identification of Escherichia albertii by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

A specific identification protocol for Escherichia albertii by using a MALDI-TOF/MS method was developed. For this purpose, a novel database was established which can differentiate E. albertii from E. coli by combining the mass spectra obtained from 58 E. albertii and 36 E. coli strains.

Oral intake of lingonberry and amla fruit extract improves skin conditions in healthy female subjects: A randomized, double-blind, placebo-controlled clinical trial.

In this study, we examined the effect of ingestion of lingonberry and amla fruit extract (LAE) on several human skin conditions. To conduct a randomized, double-blinded, placebo-controlled study, we randomly divided 99 healthy female subjects into three groups; the first group received a drink containing 25 mg of lingonberry extract and 30 mg of amla fruit extract; the second group received a drink containing double the volume of extracts received by the first group; and the third group received a placebo drink. Each participant drank 50 mL of their assigned drink once daily for 12 weeks. The primary endpoint was skin elasticity, and the secondary endpoints included skin thickness, stratum corneum water content, and degree of wrinkles around the eyes. After 12 weeks of LAE drink intake, skin elasticity showed significant, dose-dependent improvements (P < 0.01). Skin thickness, stratum corneum water content, and the degree of wrinkles also significantly improved (P < 0.001) in a dose-dependent manner. The improvements in skin elasticity and thickness, as well as in the stratum corneum water content and the degree of wrinkles, observed upon oral intake of LAE indicate that LAE may be considered a candidate anti-aging agent for preventing skin weakening.

Transdermal absorption of natural progesterone from alcoholic gel formulations with hydrophilic surfactant.

OBJECTIVES: The aim of this study was to evaluate the in vitro skin permeation and in vivo transdermal absorption of natural progesterone (Prog) from alcoholic gel-based transdermal formulations containing Prog dissolved stably at a concentration of 3%. METHODS: 3% Prog dissolved gel formulations were prepared containing with water, ethanol, 1,3-butylene glycol, carboxyvinylpolymer, diisopropanolamine, polyoxyethylene (2) oleylether and benzyl alcohol. The gel formulations added different hydrophilic surfactants and isopropyl myristate or propylene glycol dicaprylate (PGDC) as oily solvents were applied in vitro permeation study through excised rat skin on unocclusive condition. The gel formulations added polyoxyethylene (20) oleylether (Oleth-20) as hydrophilic surfactant and PGDC were applied in vivo single- and repeated-dose transdermal absorption study of rat on unocclusive condition. RESULTS: The results of evaluation of the gel formulations by an in vitro skin permeation study revealed a high flux of Prog from the formulation containing Oleth-20 and Oleth-20 with PGDC. The results of single and repeated in vivo transdermal absorption studies confirmed that good plasma levels of Prog were achieved and maintained by Oleth-20 and PGDC containing gel formulation. CONCLUSIONS: The Oleth-20 and PGDC containing ethanolic gel formulation seemed to have the ability to maintain a high activity of Prog and high diffusivity or solubility of Prog in the epidermis on the practical formulation application.

Dietary glucosylceramide enhances cornified envelope formation via transglutaminase expression and involucrin production.

In this study, we investigated whether dietary glucosylceramide (GlcCer) and its metabolite sphingoid bases, sphingosine (SS), phytosphingosine (PS), sphingadienine (SD) and 4-hydroxysphingenine (4HS), influence cornified envelope (CE) formation. CE is formed during terminal differentiation of the epidermis through crosslinking of specific precursor proteins by transglutaminases (TGases), and is essential for the skin's barrier function. Oral administration of GlcCer (0.25 mg/day) for 14 consecutive days dramatically reduced transepidermal water loss, an indicator of the skin barrier condition, in hairless mice with barrier perturbation induced by single-dose ultraviolet B (UVB) irradiation. The GlcCer treatment also increased the level of TGase-1 mRNA in UVB-irradiated murine epidermis approximately 1.6-fold compared with the control. Further, all four sphingoid bases at 1 µM concentration enhanced CE formation of cultured normal human keratinocyte cells. Among them, SS, PS and SD, but not 4HS, stimulated production of involucrin, one of the CE major precursor proteins. SD increased the expression of TGase-1 mRNA, while SS increased the expression of TGase-3 mRNA. These results indicate that the skin barrier improvement induced by oral GlcCer treatment might be at least partly due to a reinforcement of CE formation in the epidermis mediated by sphingoid bases metabolically derived from GlcCer.

Distribution and metabolism of sphingosine in skin after oral administration to mice.

In this study, (3)H- or (13)C(2),D(2)-sphingosine (SPH) was orally administered to mice to assess absorption, mass balance, tissue distribution, and metabolites in the skin. The blood concentration of (3)H-SPH showed a Tmax of 10.7 hr. The radioactivity in the skin reached 763.4 ng eq./g tissue at 12 hr, and decreased to 181.7 ng eq./g tissue at 168 hr. The concentration of radioactivity at 12 hr was 577.6 and 100.7 ng eq./g tissue in the dermis and epidermis, respectively. Thereafter, the dermis concentration decreased to 158.5 ng eq./g tissue, while the epidermis concentration increased to 298.8 ng eq./g tissue, suggesting that radioactivity moves from the dermis to the epidermis. Unchanged SPH along with lipophilic metabolites was detected in the skin of mice exposed orally to (3)H- or (13)C(2),D(2)-SPH. Moreover, in an in vitro study using human skin keratinocytes, a (13)C(2),D(2)-SPH-treatment resulted in the intracellular production of glucosylceramides (GlcCer) and ceramides (Cer) containing labeled-SPH. These results indicate the followings: first, that SPH is absorbed through the digestive tract and distributed to the skin; second, it is transferred from the dermis to the epidermis; and third, SPH is partly distributed to the skin in an unchanged form, and some of the distributed compounds are converted into GlcCer and Cer by biosynthesis.

[Underlying examination in the imaging of 89Sr bremsstrahlung radiation].

The radiopharmaceutical strontium chloride ((89)Sr) has been released as a new means of pain relief for painful bone metastasis in cancer patients. Because (89)Sr is a pure beta-emitting nuclide, it was considered difficult to know its distribution in the body from outside. Imaging with a gamma camera using bremsstrahlung radiation has been reported as one method, but there has been little detailed basic examination. We examined the optimal energy window and collimator when imaging with a gamma camera using bremsstrahlung radiation produced from (89)Sr beta rays. The results showed that setting the energy window at 75 keV, which is the peak formed by the characteristic X-ray of lead that is produced by the interaction of bremsstrahlung radiation and lead, is optimal for imaging. Also important are the material of the collimator and the use of an MELP collimator.

Distribution in skin of ceramide after oral administration to rats.

In this study, the absorption, distribution and excretion of ceramide were examined in rats. After a single oral administration of (3)H-ceramide, mean plasma concentration of radioactivity reached maximum at approximately 10.67 hr and decreased with a T(1/2) of 67.12 hr. The mean cumulative excretion of radioactivity in urine and feces accounted for approximately 4.79% and 87.44%, respectively, of the dose. At 96 hr after dosing, 1.67% and 3.67%, respectively, of the dose were still present in the skin and carcass. The radioactivity in the skin at 12 hr was lower than that in plasma and the ratio of skin to plasma concentration was 0.7. However, at 120 hr after dosing, the ratio of skin to plasma concentration increased to 4. A detailed analysis of the distribution of radioactivity in a section of skin showed that radioactivity was located in the dermis and epidermis. At 168 hr, the radioactivity in the epidermis was 8.0% of the radioactivity in skin. The results of the present study clearly demonstrate that some ceramide orally administered is distributed gradually in the dermis after gastrointestinal absorption, followed by transfer from the dermis to the epidermis.