Swim-up method is superior to density gradient centrifugation for preserving sperm DNA integrity during sperm processing.

Purpose: This study aimed to evaluate the effects of swim-up and density gradient centrifugation methods on sperm DNA fragmentation. Methods: Nineteen normozoospermic patient samples with ≥100 × 106 motile sperms were included in this study. Sperm DNA fragmentation, progressive motility, and progressive motile sperm number were measured before and after the swim-up method or density gradient centrifugation. Results: Sperm DNA fragmentation was not statistically different between swim-up-(14.4 ± 2.1%, p = 0.32) and density gradient centrifugation-processed (25.0 ± 3.0%, p = 0.20) and unprocessed semen samples (19.2 ± 1.9%). Sperm DNA fragmentation was significantly lower in swim-up-than in density gradient centrifugation-processed samples (p < 0.05). Sperm progressive motility was significantly higher (p < 0.05) in swim-up-(92.9 ± 1.0%) and density gradient centrifugation-processed (81.3 ± 2.0%) samples, with the former being higher, than in unprocessed semen samples (53.1 ± 3.7%). The recovery rate of progressive motile sperms was significantly lower in swim-up-(9.7 ± 1.4%) than in density gradient centrifugation-processed samples (17.2 ± 1.8%, p < 0.05). Conclusions: The swim-up method is superior to density gradient centrifugation, evidenced by less sperm DNA fragmentation and higher sperm progressive motility. The recovery rate of progressive motile sperms was better after density gradient centrifugation than after swim-up.

Co-Self-Assembly of Amphiphiles into Nanocomposite Hydrogels with Tailored Morphological and Mechanical Properties.

As one of the most amazing aspects of life, all living organisms are formed by self-assembly, a fundamental biological design process in which ordered nanostructures are assembled from small parts. For example, most of the biological tissues contain structurally soft and hard parts that are usually hierarchically organized at nano or micro levels to achieve specific functions. Hydrogels are one of the most promising soft materials owing to their potential applications in building of biological tissues and stretchable sensors. In this work, a series of hydrogels are synthesized through the co-self-assembly of two types of amphiphiles in their aqueous solution prior to polymerization. Soft and hard parts with nanostructures of different order parameters are incorporated into the hydrogels. The hydrophilic segment (as soft phases) of the polymer network provides water absorption, fluid flow, and softness, whereas the hydrophobic segment (as hard phases) provides strength and tearing and fracture resistance. Appropriate soft/hard nanostructures and their interfaces allow for the tailoring of the desired morphological and mechanical properties, including a different wetting ability, toughness, energy dissipation, self-recovery, and fracture resistance arising from their nanostructures. This work provides insights into the design of nanostructured anisotropic hydrogels with controlled morphological and mechanical properties.

Heterotopic pregnancy with suspicion of superfetation after the intrauterine insemination cycle with ovulation induction using clomiphene citrate: A case report.

At 22 days after intrauterine insemination with ovulation induction using clomiphene citrate at a previous hospital, a 30-year-old woman was admitted to our hospital owing to right lower quadrant abdominal pain. We diagnosed threatened abortion because of a gestational sac in the uterus on transvaginal ultrasonography. The next day, she complained of increased abdominal pain. Transvaginal ultrasonography revealed a gestational sac-like structure in the echo free space. She was diagnosed with heterotopic pregnancy due to a ruptured right tubal pregnancy, underwent laparoscopic right salpingectomy. Rupture of a gestational age of 5 weeks is extremely rare. If this was a case of a heterotopic pregnancy due to superfetation, it could be explained as this clinical course. When a pregnant woman develops abdominal pain, heterotopic pregnancies should not be excluded from the differential diagnosis, and the possibility of superfetation should be taken into consideration.

Fluorescence via Reverse Intersystem Crossing from Higher Triplet States in a Bisanthracene Derivative.

To elucidate the high external quantum efficiency observed for organic light-emitting diodes using a bisanthracene derivative (BD1), non-radiative transition processes as well as radiative ones are discussed employing time-dependent density functional theory. It has been previously reported that the observed high external quantum efficiency of BD1 cannot be explained by the conventional thermally activated delayed fluorescence involving T1 exciton nor triplet-triplet annihilation. The calculated off-diagonal vibronic coupling constants of BD1, which govern the non-radiative transition rates, suggest a fluorescence via higher triplets (FvHT) mechanism, which entails the conversion of a high triplet exciton generated during electrical excitation into a fluorescent singlet exciton. This mechanism is valid as long as the relaxation of high triplet states to lower states is suppressed. In the case of BD1, its pseudo-degenerate electronic structure helps the suppression. A general condition is also discussed for the suppression of transitions in molecules with pseudo-degenerate electronic structures.

Bimolecular Rate Constants for FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus and Organic Electron Acceptors.

The flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) from Aspergillus species require suitable redox mediators to transfer electrons from the enzyme to the electrode surface for the application of bioelectrical devices. Although several mediators for FAD-GDH are already in use, they are still far from optimum in view of potential, kinetics, sustainability, and cost-effectiveness. Herein, we investigated the efficiency of various phenothiazines and quinones in the electrochemical oxidation of FAD-GDH from Aspergillus terreus. At pH 7.0, the logarithm of the bimolecular oxidation rate constants appeared to depend on the redox potentials of all the mediators tested. Notably, the rate constant of each molecule for FAD-GDH was approximately 2.5 orders of magnitude higher than that for glucose oxidase from Aspergillus sp. The results suggest that the electron transfer kinetics is mainly determined by the formal potential of the mediator, the driving force of electron transfer, and the electron transfer distance between the redox active site of the mediator and the FAD, affected by the steric or chemical interactions. Higher k2 values were found for ortho-quinones than for para-quinones in the reactions with FAD-GDH and glucose oxidase, which was likely due to less steric hindrance in the active site in the case of the ortho-quinones.

A genetic link between epigenetic repressor AS1-AS2 and a putative small subunit processome in leaf polarity establishment of Arabidopsis.

Although the DEAD-box RNA helicase family is ubiquitous in eukaryotes, its developmental role remains unelucidated. Here, we report that cooperative action between the Arabidopsis nucleolar protein RH10, an ortholog of human DEAD-box RNA helicase DDX47, and the epigenetic repressor complex of ASYMMETRIC-LEAVES1 (AS1) and AS2 (AS1-AS2) is critical to repress abaxial (ventral) genes ETT/ARF3 and ARF4, which leads to adaxial (dorsal) development in leaf primordia at shoot apices. Double mutations of rh10-1 and as2 (or as1) synergistically up-regulated the abaxial genes, which generated abaxialized filamentous leaves with loss of the adaxial domain. DDX47 is part of the small subunit processome (SSUP) that mediates rRNA biogenesis. In rh10-1 we found various defects in SSUP-related events, such as: accumulation of 35S/33S rRNA precursors; reduction in the 18S/25S ratio; and nucleolar hypertrophy. Double mutants of as2 with mutations of genes that encode other candidate SSUP-related components such as nucleolin and putative rRNA methyltransferase exhibited similar synergistic defects caused by up-regulation of ETT/ARF3 and ARF4 These results suggest a tight link between putative SSUP and AS1-AS2 in repression of the abaxial-determining genes for cell fate decisions for adaxial development.

Effect of dalteparin, a low-molecular-weight heparin, as adjunctive therapy in patients with Kawasaki disease: a retrospective study.

BACKGROUND: Dalteparin, a low-molecular-weight heparin, has anticoagulant and anti-angiogenic activity. This study investigated whether dalteparin reduced coronary artery lesion (CAL) prevalence, and resistance to intravenous immunoglobulin (IVIG) therapy in Kawasaki disease (KD). METHODS: This retrospective study comprised two parts. In the first cohort, 126 patients with KD (68 male, 58 female; median age: 22 months, range: 1-67 months) admitted to Nihon University Nerima-Hikarigaoka Hospital from January 2004 to June 2008, received either dalteparin 75 IU/kg/day, IVIG 400 mg/kg/day for 5 consecutive days, and aspirin 30 mg/kg/day, or dalteparin 75 IU/kg/day and aspirin 30 mg/kg/day, until clinical improvement. Control data came from the 2005-6 Nationwide KD survey. In the second cohort, 112 patients with KD (59 male, 53 female; median age: 19 months, range: 1-66 months) admitted from June 2010 to February 2012, received either dalteparin 75 IU/kg/day, IVIG 2.0 g/kg over 12 h, and aspirin 30 mg/kg/day, or dalteparin 75 IU/kg/day and aspirin 30 mg/kg/day. Control data came from the 2009-10 Nationwide KD survey. No patients enrolled in the nationwide surveys received dalteparin. All patients at our institution were given dalteparin in their combination therapy. RESULTS: A comparison of the first cohort with controls in the nationwide survey showed that the prevalence of initial administration of IVIG was 80.2% versus 86.0%; the rate of additional IVIG administration was 7.1% versus 14.0% (p = 0.03); CAL prevalence in the acute period was 4.8% versus 11.9% (p < 0.01); and the prevalence of cardiovascular sequelae was 0% versus 3.8% (p < 0.05). A comparison of the second cohort with controls in the nationwide survey showed that the rate of initial administration of IVIG was 92.9% versus 89.5%; the rate of additional IVIG administration was 8.9% versus 17.1% (p = 0.02); the prevalence of resistance to IVIG was 3.6% versus 14.9% (p < 0.001); and CAL prevalence in the acute period was 2.7% versus 8.6% (p = 0.03). CONCLUSIONS: This study found that adjunctive dalteparin was associated with a lower prevalence of IVIG resistance and CAL in young children with KD. TRIAL REGISTRATION UMIN-CTR: UMIN000010349.

Serum vitamin D concentrations and associated severity of acute lower respiratory tract infections in Japanese hospitalized children.

BACKGROUND: Vitamin D is an immunomodulatory molecule related to innate immunity that may contribute to the increased occurrence of acute lower respiratory infection (ALRI) in children, one of the most common reasons for hospitalization and intensive care unit admission. In the present study, the association between vitamin D deficiency and the severity of respiratory infection was evaluated by determining serum concentrations of 25-hydroxyvitamin D (25(OH)D) in a group of hospitalized children with ALRI. METHODS: Of the 28 children admitted to Nihon University Nerima-Hikarigaoka Hospital with ALRI over the period November 2008­May 2009, 26 were diagnosed as having bronchiolitis and two were found to have pneumonia. A competitive protein binding radioimmunoassay was used to determine serum 25(OH)D concentrations. RESULTS: Mean 25(OH)D concentrations in breast-fed children with ALRI (n = 7) were significantly lower than those in children with ALRI who were bottle fed/weaned (n = 6) or on a regular diet (n = 15; 14.6 ± 9.7, 28.9 ± 6.9 and 24.6 ± 8.8 ng/mL, respectively). There was a significant correlation between vitamin D deficiency (<15 ng/mL) and the need for supplementary oxygen and ventilator management. CONCLUSION: Significantly more children with ALRI who needed supplementary oxygen and ventilator management were vitamin D deficient. These findings suggest that the immunomodulatory properties of vitamin D may influence the severity of ALRI.

Erythema nodosum associated with Yersinia enterocolitica infection.

We report a 74-year-old woman who presented to hospital with fever, vomiting, diarrhea, and 2 weeks later developed erythema nodosum (EN) on the legs, and was diagnosed with Yersinia enterocolitica infection based on her clinical course and microbiological examination of the stool. She also had a complication of pancreatitis, which made the diagnosis challenging. We should suspect infection by Y. enterocolitica when diagnosing cases of EN with gastrointestinal symptoms. We assume EN is likely to appear 2 weeks after the onset of gastrointestinal symptoms from our case and other case reports.

Serum protein adsorption and platelet adhesion on aspartic-acid-immobilized polysulfone membranes.

Polysulfone (PSf) membranes that covalently conjugated with aspartic acid (ASP-PSf) were prepared and analyzed for hemocompatability. Compared to PSf or other types of surface-modified PSf membranes, the ASP-PSf membranes had a reduced ability to adsorb protein from either a plasma solution or a mixed solution of albumin, globulin and fibrinogen. This appears to be due to the creation of a hydrophilic surface by the aspartic acid zwitterion immobilized on the ASP-PSf membranes. Furthermore, the analyses of membrane protein adsorption showed that a mixed protein solution recapitulates the cooperative adsorption of proteins that occurs in plasma. We also found that the number of adhering platelets was the lowest on the ASP-PSf membranes and, in general, that platelet adhesion decreased in parallel with fibrinogen adsorption. In summary, aspartic acid immobilized on the ASP-PSf membranes, which have zwitterions with a net zero charge, effectively contributes to the hydrophilic and hemocompatible sites on the surface of the hydrophobic PSf membranes.