Hydrodynamic evaluation of a new dispersive aortic cannula (Stealthflow).

The aim of this study was to evaluate flow from a new dispersive aortic cannula (Stealthflow) in the aortic arch using flow visualization methods. Particle image velocimetry was used to analyze flow dynamics in the mock aortic model. Flow patterns, velocity distribution, and streamlines with different shape cannulas were evaluated in a glass aortic arch model. We compared flow parameters in two different dispersive type cannulas: the Stealthflow and the Soft-flow cannula. A large vortex and regurgitant flow were observed in the aortic arch with both cannulas. With the Stealthflow cannula, a high-velocity area with a maximum velocity of 0.68 m/s appeared on the ostium of the cannula in the longitudinal plane. With the Soft-flow cannula, 'multiple jet streams, each with a velocity less than 0.60 m/s, were observed at the cannula outlet. Regurgitant flow from the cannula to the brachiocephalic artery and to the ascending aorta on the greater curvature was specific to the Soft-flow cannula. The degree of regurgitation on the same site was lower with the Stealthflow cannula than with the Soft-flow cannula. The Stealthflow cannula has similar flow properties to those of the Soft-flow cannula according to glass aortic model analysis. It generates gentle flow in the aortic arch and slow flow around the ostia of the aortic arch vessels. The Stealthflow cannula is as effective as the Soft-flow cannula. Care must be taken when the patient has thick atheromatous plaque or frail atheroma on the lesser curvature of the aortic arch.

Cannulation needle-induced anterior wall tenting of internal jugular vein causing posterior wall penetration.

Unintentional posterior venous wall penetration during internal jugular vein (IJV) cannulation may cause critical arterial injuries in spite of ultrasound guidance. We aimed to evaluate whether small venous diameter and anterior venous wall tenting by a needle would be associated with posterior venous wall penetration, and to seek factors related to the venous wall tenting. We conducted a retrospective review in patients who underwent IJV cannulation. Using an ultrasound view obtained when puncturing, venous diameter, venous wall thickness, anterior venous wall tenting length, and needle angle were measured, and posterior venous wall penetration was determined. Eleven cannulations in 56 patients were assigned to posterior venous wall penetration. Small venous diameter (p = 0.004), and long anterior venous wall tenting (p = 0.007) were associated with posterior venous wall penetration. The longer anterior venous tenting would be expected with reducing needle angle (p = 0.004) or increasing anterior venous wall thickness (p = 0.006). In conclusion, small IJV and anterior venous wall tenting lead to posterior venous wall penetration. Anterior venous wall tenting is longer with reducing needle angle, or increasing the anterior venous wall thickness.

A report from Fukushima: an assessment of bone health in an area affected by the Fukushima nuclear plant incident.

Bone health was assessed for inhabitants of an area affected by the Fukushima nuclear plant incident. Osteoporotic patients, who had been treated with active vitamin D3 and/or bisphosphonate at Soma Central Hospital before the Fukushima incident, were enrolled. Changes in bone turnover markers and bone mineral density were retrospectively analyzed. Serum levels of a bone resorption marker, serum type I collagen cross-linked N-telopeptide were decreased in all the treated groups, whereas those of a bone formation marker, bone-specific alkaline phosphatase, were increased. Accordingly, bone mineral density, estimated by dual-energy X-ray absorptiometry, was increased in the lumbar spine of all groups, but bone mass increase in the proximal femur was detected only in the group treated with the two agents in combination. From the degree of these parameter changes, the antiosteoporotic treatments looked effective and were equivalent to the expected potency of past observations. At this stage, the present study implies that the Fukushima nuclear incident did not bring an acute risk to bone health in the affected areas.

Prevalence and antimicrobial susceptibility of foodborne bacteria in wild boars (Sus scrofa) and wild deer (Cervus nippon) in Japan.

This study aimed to evaluate the role of wild boars and deer as reservoirs of foodborne bacteria. We investigated the prevalence and antimicrobial susceptibility of Campylobacter spp., Salmonella spp., Shiga toxin-producing Escherichia coli (STEC) O157 and O26, and Listeria monocytogenes isolated from wild boars and deer in Japan, from July through December 2010. Campylobacter spp. and Salmonella spp. were isolated from 43.8% (95% confidence interval [CI]: 35.0-52.6) and 7.4% (95% CI: 2.8-12.1) of rectal content samples of wild boars, respectively, but not from wild deer. The most common Campylobacter species was C. lanienae and C. hyointestinalis. The nine Salmonella serovars isolated were S. enterica subsp. enterica serovar Agona (three isolates), S. Narashino (two), S. Enteritidis (one), S. Havana (one), S. Infantis (one), and S. Thompson (one). Five (16%) and 6 (29%) isolates of C. lanienae and C. hyointestinalis, respectively, were resistant to enrofloxacin. STEC O157 and O26 and L. monocytogenes were isolated from 2.3% (95% CI: 0-5.0), 0.8% (95% CI: 0-2.3), and 6.1% (95% CI: 1.7-10.5) of the rectal content samples of wild deer, respectively, but not from wild boars. This first nationwide survey of the prevalence of foodborne bacteria in wild boars and wild deer in Japan suggests that consumption of meat from these animals is associated with the risk of causing infection with these bacteria in humans. Moreover, these animals are potential vehicles for distribution of antimicrobial-resistant bacteria into their habitat. The prevalence and antimicrobial susceptibility of such foodborne bacteria in these wild animals should be monitored periodically.

Gene expression of type II collagen is regulated by direct interaction with Kruppel-like factor 4 and AT-rich interactive domain 5B.

We have previously found and characterized two pairs of enhancer elements, E1 and E2, in the type II collagen alpha 1 chain (COL2A1) gene. Subsequent studies have suggested that these enhancers function differently in the regulation of gene expression. For example, histone deacetylase 10 modifies only the E2 enhancer region to affect gene expression. Therefore, in this study, we aimed to clarify the transcriptional complex formed at each enhancer region by identifying transcription factors that specifically bind to each enhancer element. To this end, we used chondrocytic cell lines established using our unique silent reporter system and overexpressed candidate transcription factors in these cells. We found two transcription factors, other than the SOX trio, that directly bound to COL2A1 and regulated its expression. The first was Kruppel-like factor-4 (KLF4), which bound to the promoter proximal region, and the second was AT-rich interactive domain 5B (ARID5B) which bound to the E1 enhancer element. Further studies are needed to identify factors that specifically bind to the E2 enhancer element. In any case, our findings provide an important insight into the molecular mechanisms underlying the regulation of COL2A1. In this paper, we reevaluated the previous analysis of transcription factors involved in the regulation of COL2A1 expression.

Purification, characterization and molecular cloning of a novel endo-beta-N-acetylglucosaminidase from the basidiomycete, Flammulina velutipes.

Endo-beta-N-acetylglucosaminidases are thought to be key enzymes in the catabolism of asparagine-linked oligosaccharides. However, little is known about the enzymes of this type in basidiomycetes. We investigated endo-beta-N-acetylglucosaminidases in basidiomycetes using fluorescence-labeled glycoasparagines as substrates. Flammulina velutipes showed high activity and its enzyme was named endo-beta-N-acetylglucosaminidase FV (Endo FV). The enzyme purified from the fruiting bodies of F. velutipes was separated into two forms. Endo FV was specific for high mannose and hybrid-type oligosaccharides. The enzyme was remarkably less active against asparagine-linked oligosaccharides attached to glycoproteins. It transferred an asparagine-linked oligosaccharide to Glc, but not to Gal. cDNA of Endo FV was cloned. It was composed of a 996-bp open reading frame encoding 331 amino acid residues. A recombinant Endo FV expressed in Escherichia coli showed enzymatic activity. The Endo FV gene in the genome of F. velutipes had no introns. The gene encoding Endo FV showed little homology with genes of known endo-beta-N-acetylglucosaminidases. A chitinase active site motif existed in the deduced primary structure, indicating that Endo FV belongs to glycoside hydrolase family 18. The deduced amino acid sequence of Endo FV had regions conserved in class III chitinases from fungi though it showed little homology with the sequence of any other endo-beta-N-acetylglucosaminidases. A folding model of Endo FV indicated it to be homologous with the tertiary structure of Endo H which is quite similar in specificity for asparagine-linked oligosaccharides. This study suggests that Endo FV may become similar to Endo H in substrate specificity as a result of evolutionary convergence.

Snapping elbow with congenital radial head dislocation: case report.

An 11-year-old boy with congenital radial head dislocation experienced painful snapping of his left elbow upon movement. He had no previous history of trauma. A plain radiograph of his left elbow showed anterior dislocation of the radial head and flexion deformity of the hypoplastic radial neck. Arthroscopy showed that the snapping of the elbow occurred between the annular ligament and the dislocated radial head during elbow flexion and extension. After the annular ligament was released, the snapping immediately disappeared. Five years after the surgery, the patient has no pain or snapping upon elbow movement.

Levansucrase from Bacillus amyloliquefaciens KK9 and Its Y237S Variant Producing the High Bioactive Levan-Type Fructooligosaccharides.

Levan-typed fructooligosaccharide (LFOS), a ß-2,6 linked oligofructose, displays the potential application as a prebiotic and therapeutic dietary supplement. In the present study, LFOS was synthesized using levansucrase from Bacillus amyloliquefaciens KK9 (LsKK9). The wild-type LsKK9 was cloned and expressed in E. coli, and purified by cation exchanger chromatography. Additionally, Y237S variant of LsKK9 was constructed based on sequence alignment and structural analysis to enhance the LFOS production. High-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) analysis indicated that Y237S variant efficiently produced a higher amount of short-chain LFOS than wild type. Also, the concentration of enzyme and sucrose in the reactions was optimized. Finally, prebiotic activity assay demonstrated that LFOS produced by Y237S variant had higher prebiotic activity than that of the wild-type enzyme, making the variant enzyme attractive for food biotechnology.

Plasticity and regulatory mechanisms of Hox gene expression in mouse neural crest cells.

In amniotes, the developmental potentials of neural crest cells differ between the cranium and the trunk. These differences may be attributable to the different expression patterns of Hox genes between cranial and trunk neural crest cells. However, little is known about the factors that control Hox genes expression in neural crest cells. The present data demonstrate that retinoic acid (RA) treatment and the activation of Wnt signaling induce Hoxa2 and Hoxd9 expression, respectively, in mouse mesencephalic neural crest cells, which never express Hox genes in vivo. Furthermore, Wnt signaling suppresses the induction of Hoxa2. We also demonstrate that these factors participate in the maintenance of Hoxa2 and Hoxd9 expression in mouse trunk neural crest cells. Our results suggest that RA and Wnt signaling function as environmental factors that regulate the expression of Hoxa2 and Hoxd9 in mouse neural crest cells.

Spatio-temporal expression of activating transcription factor 5 in the skeletal development of mouse limb.

There are no data about the expression pattern of activating transcription factor 5 (Atf5) during limb development. The aim of the present study was to describe the expression of Atf5 throughout mouse limb development. Some specimens were examined for the expression of type II collagen alpha1 (Col2a1), sex-determining region Y-related high-mobility-group-box 9 (Sox9), and activating transcription factor 2 (Atf2) by in situ hybridization. ATDC5 cells, mouse limb buds, and epiphyseal cartilage were examined for the expression of Atf5alpha and Atf5beta by quantitative real-time PCR. Atf5 transcripts were detected in condensed mesenchymal cells, cartilage primordia, proliferative, prehypertrophic, and articular chondrocytes, but not in hypertrophic chondrocytes. Atf5 was predominantly expressed by osteoblasts in the primary spongiosa. At the postnatal stage, Atf5 was expressed in epiphyseal chondrocytes and osteoblasts lining the bone trabeculae. Spatial distribution of Col2a1, Sox9, and Atf2 transcripts closely corresponded to that of Atf5 within developing limbs, except for bony tissues. In cartilage, Atf5alpha is the dominant form in the embryonic stage as well as in the postnatal stage.

De Quervain disease caused by abductor pollicis longus tenosynovitis: a report of three cases.

De Quervain disease is caused by a stenosing tenosynovitis in the first dorsal compartment, and the main aetiology is extensor pollicis brevis (EPB) tenosynovitis. We encountered three cases in which EPB tenosynovitis was absent and abductor pollicis longus (APL) tenosynovitis was confirmed during operation. In the treatment of de Quervain disease, APL tenosynovitis should be paid as much attention as EPB tenosynovitis.

Production and purification of mannan oligosaccharide with epithelial tight junction enhancing activity.

BACKGROUND: Mannanan oligosaccharide (MOS) is well-known as effective supplement food for livestock to increase their nutrients absorption and health status, but the structure and identification of bioactive MOS remain unclear. In this study, MOS production was accomplished, using enzymatic hydrolysis of pretreated coconut meal substrate with recombinant mannanase. METHODS: The mannanase gene was cloned from Bacillus subtilis cAE24, then expressed in BL21. Purified Mannanase exhibit stability over a wide range of pH and temperature from pH 6-8 and 4 °C to 70 °C, respectively. SEM analysis revealed that sonication could change the surface characteristic of copra meal, which gave better MOS yield, compared to untreated substrates. The separation and purification of each MOS were achieved using Biogel-P2 column chromatography. Determination of biological active MOS species was also investigated. T84 cells were cultured and treated with each of the purified MOS species to determine their tight junction enhancing activity. RESULTS: Scanning electron microscope imaging showed that pretreatment using sonication could disrupt the surface of copra meal better than grinding alone, which can improve the production of MOS. Pentamer of MOS (M5) significantly increased tight junction integration of T84 cells measured with TEER (p < 0.0001).

Galactomannan Pentasaccharide Produced from Copra Meal Enhances Tight Junction Integration of Epithelial Tissue through Activation of AMPK.

Mannan oligosaccharide (MOS) is well-known as an effective fed supplement for livestock to increase their nutrients absorption and health status. Pentasaccharide of mannan (MOS5) was reported as a molecule that possesses the ability to increase tight junction of epithelial tissue, but the structure and mechanism of action remains undetermined. In this study, the mechanism of action and structure of MOS5 were investigated. T84 cells were cultured and treated with MOS5 compared with vehicle and compound C, a 5'-adenosine monophosphate-activated protein kinase (AMPK) inhibitor. The results demonstrated that the ability of MOS5 to increase tight junction integration was inhibited in the presence of dorsomorphine (compound C). Phosphorylation level of AMPK was elevated in MOS5 treated group as determined by Western blot analysis. Determination of MOS5 structure was performed using enzymatic mapping together with 1H, 13C NMR, and 2D-NMR analysis. The results demonstrated that the structure of MOS5 is a ß-(1,4)-mannotetraose with α-(1,6)-galactose attached at the second mannose unit from non-reducing end.

Notch signaling is required for the chondrogenic specification of mouse mesencephalic neural crest cells.

We examined the roles of Notch signaling in the chondrogenesis of mouse mesencephalic neural crest cells. The present study demonstrated that the activation of Notch signaling or the treatment with fibroblast growth factors (FGFs) promotes the differentiation of proliferative and prehypertrophic chondrocytes expressing collagen type II. Notch activation or FGF2 exposure during the first 24h in culture was critical for the differentiation of proliferative and prehypertrophic chondrocytes. The expression of SOX9, a transcription activator of collagen type II, was also upregulated by Notch activation or FGF2 treatment. The promotion of proliferative and prehypertrophic chondrocyte differentiation by FGF2 was significantly suppressed by the inhibition of Notch signaling using Notch-1 siRNA. These results suggest that FGFs activate Notch signaling and that this activation promotes the chondrogenic specification of mouse mesencephalic neural crest cells. Furthermore, we investigated the expression patterns of Notch-1, SOX9, and p75, which is a marker of undifferentiated neural crest cells, in the mandibular arch where mesencephalic neural crest cells colonize and undergo chondrogenesis. These in vivo observations, coupled with the results of the present in vitro study, suggest that Notch signaling as well as FGFs is a component of epithelial-mesenchymal interactions that promote the chondrogenic specification of mouse mesencephalic neural crest cells.

Convenient preparation and characterization of a monoclonal antibody for the N-linked sugar chain of a glycoprotein using a microbial endoglycosidase.

We attempted to obtain the monoclonal antibody specific for the N-linked complex-type sialo-oligosaccharide in glycoproteins. We first synthesized a chimeric immunoantigen having an N-linked complex-type of oligosaccharide of glycopeptide, which was bound to a p-formylphenyl compound and conjugated with phosphatidylethanolamine dimyristoyl using the transglycosylation activity of a microbial endoglycosidase (Endo-M) and a reductive amination reaction. This preparative method was convenient and provided a good yield. By immunizing mice with this chimeric neoglycolipid, the monoclonal antibody for the complex-type of sialo-oligosaccharide was obtained in the culture fluid of the cell line even though it was relatively unstable. The monoclonal antibody reacted with various glycoproteins having complex-type sialo-oligosaccharides, but not with those having complex-type asialo-oligosaccharides and high mannose types of oligosaccharides, or with any glycosphingolipids. One of epitopes of this monoclonal antibody seemed to be an alpha-2,6-linked sialic acid at the non-reducing end of the sialo-oligosaccharide of the glycoprotein.

Nilotinib vs. imatinib in Japanese patients with newly diagnosed chronic myeloid leukemia in chronic phase: long-term follow-up of the Japanese subgroup of the randomized ENESTnd trial.

In the ongoing, international, phase 3 study Evaluating Nilotinib Efficacy and Safety in Clinical Trials-Newly Diagnosed Patients (ENESTnd), nilotinib 300 and nilotinib 400 mg, both twice daily, are compared with imatinib 400 mg once daily for the treatment of newly diagnosed chronic myeloid leukemia in the chronic phase (CML-CP). Results for the overall population in ENESTnd (n = 846) showed that nilotinib resulted in higher response rates vs. imatinib and was well tolerated. Outcomes among Japanese patients in ENESTnd were specifically analyzed after 1 year of follow-up, and showed similar trends to the overall population; we present updated analysis of the Japanese subgroup based on 5 years of follow-up. Among Japanese patients in the nilotinib 300-mg (n = 29), nilotinib 400-mg (n = 23), and imatinib (n = 25) arms, 86.2, 78.3, and 60.0%, respectively, achieved major molecular response [BCR-ABL1 ≤ 0.1% on the International Scale (BCR-ABL1 IS)] by 5 years, and 65.5, 69.6, and 40.0%, respectively, achieved MR4.5 (BCR-ABL1 IS ≤ 0.0032%). Safety results were consistent with prior reports. In this subgroup, one death occurred during treatment in the nilotinib 400-mg twice-daily arm (unknown cause), and one patient in each arm progressed to accelerated phase/blast crisis by the data cutoff.

Ruxolitinib is effective and safe in Japanese patients with hydroxyurea-resistant or hydroxyurea-intolerant polycythemia vera with splenomegaly.

Ruxolitinib, a potent JAK1/JAK2 inhibitor, was found to be superior to the best available therapy (BAT) in controlling hematocrit, reducing splenomegaly, and improving symptoms in the phase 3 RESPONSE study of patients with polycythemia vera with splenomegaly who experienced an inadequate response to or adverse effects from hydroxyurea. We report findings from a subgroup analysis of Japanese patients in RESPONSE (n = 18). The composite response rate (hematocrit control and spleen response) was higher in patients receiving ruxolitinib (50.0%) than in those receiving BAT (8.3%). A total of 50.0% of patients randomized to ruxolitinib achieved a spleen response vs 8.3% of those receiving BAT; 100 and 33.3% of patients in the respective groups achieved hematocrit control, with mean hematocrit in ruxolitinib-treated patients remaining stable at < 45% throughout the study. Similarly, a higher proportion of ruxolitinib-treated patients achieved complete hematologic remission (33.3 vs 16.7%). Ruxolitinib also led to rapid improvements in pruritus. All responses with ruxolitinib were durable to week 80, and its safety profile was consistent with that in the overall study. These findings suggest that ruxolitinib is an effective and well-tolerated treatment option for Japanese patients with polycythemia vera with an inadequate response to or adverse effects from hydroxyurea.

[Therapeutic agents for disorders of bone and calcium metabolism--Denosumab, a fully human monoclocal antibody-targeting RANKL as a therapy for cancer-induced bone diseases].

Receptor activator of nuclear-kappaB ligand (RANKL) is a protein expressed by oseoblastic stromal cells, binds to receptor activator of nuclear factor-kappaB (RANK) and is the primary mediator of osteoclast differentiation, activation, and survival. RANKL is responsible for osteoclast-mediated bone resorption in a broad range of conditions and play a key role in establishment and propagation of skeletal disease in patients with advanced cancer. Denosumab is a fully human monoclonal antibody that binds RANKL with high affinity and specificity and inhibits RANKL-RANK interaction, mimicking the endogenous effects of osteoprotegerin, a soluble RANKL decoy receptor. In the phase 1 clinical trials in healthy post menopausal women and patients with multiple myeloma or breast cancer with bone metastasis including Japanese (except for multiple myeloma) showed that single and multiple subcutaneous injection of denosumab caused rapid and sustained suppression of markers of osteoclastic bone resorption with favorable safety profiles. Currently, the larger global clinical trials to investigate the effect of this agent for the treatment of cancer-induced bone disease as well as osteoporosis are underway.

Mouse embryonic dorsal root ganglia contain pluripotent stem cells that show features similar to embryonic stem cells and induced pluripotent stem cells.

In the present study, we showed that the dorsal root ganglion (DRG) in the mouse embryo contains pluripotent stem cells (PSCs) that have developmental capacities equivalent to those of embryonic stem (ES) cells and induced pluripotent stem cells. Mouse embryonic DRG cells expressed pluripotency-related transcription factors [octamer-binding transcription factor 4, SRY (sex determining region Y)-box containing gene (Sox) 2, and Nanog] that play essential roles in maintaining the pluripotency of ES cells. Furthermore, the DRG cells differentiated into ectoderm-, mesoderm- and endoderm-derived cells. In addition, these cells produced primordial germ cell-like cells and embryoid body-like spheres. We also showed that the combination of leukemia inhibitor factor/bone morphogenetic protein 2/fibroblast growth factor 2 effectively promoted maintenance of the pluripotency of the PSCs present in DRGs, as well as that of neural crest-derived stem cells (NCSCs) in DRGs, which were previously shown to be present there. Furthermore, the expression of pluripotency-related transcription factors in the DRG cells was regulated by chromodomain helicase DNA-binding protein 7 and Sox10, which are indispensable for the formation of NCSCs, and vice versa. These findings support the possibility that PSCs in mouse embryonic DRGs are NCSCs.

Posteromedial papillary muscle rupture due to squeezing of the left anterior descending coronary artery.

An unusual case of posteromedial papillary muscle (PPM) rupture due to isolated left anterior descending (LAD) artery ischaemia, associated with severe myocardial bridge contraction, is presented. The unusual blood supply to the PPM was associated with its apical origin and apex-forming LAD.