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Objectives: Gene therapy using viral vectors and antibody-based therapies continue to expand within the pharmaceutical market. We evaluated whether Cellhesion ® VP, a chitin-based material, can be used as 3D culture platform for cell lines used for the production of antibodies and viral vectors., Results: The results of Cell Counting Kit-8 assay and LDH assay revealed that Cellhesion ® VP had no adverse effect to Human Embryonic Kidney (HEK) 293, A549 and Chinese hamster ovary (CHO) DG44-Interferon-β (IFN) cells. Cell growth analyses showed that Cellhesion ® VP supported the 3D culture of HEK293, A549 and CHO DG44- IFN-β cells with a spherical morphology. Importantly, subculture of these cell lines on Cellhesion ® VP was easily performed without trypsinization because cells readily transferred to newly added scaffold. Our data also suggest that CHO DG44-IFNβ, cultured on Cellhesion ® VP secreted IFNβ stably and continuously during the culture period., Conclusions: Cellhesion ® VP provides a simple and streamlined expansion culture system for the production of biopharmaceuticals., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Mesenchymal stem cell (MSC) transplantation, in particular allogeneic transplantation, is a promising therapy for a variety of diseases. However, before performing allograft treatment it is necessary to find suitable donors, establish culture methods that maintain cell quality, and reduce cell production costs. Here, we present a new method of producing allogeneic MSCs combining human umbilical cord-derived mesenchymal stem cells (UCMSCs) and chitin-based polysaccharide fibers (Cellhesion ® MS). UCMSC numbers significantly increased, and cells grew as dispersed spheres on Cellhesion ® MS. Subsequent biological analyses showed that the expression levels of stemness-related and migration-related genes were significantly upregulated, including octamer-binding transcription factor 4 (OCT4), Nanog homeobox (NANOG), and C-X-C chemokine receptor type 4 (CXCR4). The secretion levels of paracrine factors such as prostaglandin E2 (PGE 2 ), TNFα-stimulating gene (TSG)-6, fibroblast growth factor 2 (bFGF), and Angiogenin (Ang) from UCMSCs using Cellhesion ® MS were significantly higher than with microcarrier and U-bottom plate culture. In addition, culture supernatant from UCMSCs with Cellhesion ® MS had better angiogenic potential than that from monolayer cultured UCMSCs. Furthermore, we succeeded in a scaled-up culture of UCMSCs with Cellhesion ® MS using a closed culture bag. Therefore, Cellhesion ® MS is a key material for producing high-quality UCMSCs in a three-dimensional (3D) culture system.

Signal-transducing adaptor family member-2 (STAP-2) is an adaptor protein that regulates various intracellular signaling pathways and promotes tumorigenesis in melanoma and breast cancer cells. However, the contribution of STAP-2 to the behavior of other types of cancer cells is unclear. Here, we show that STAP-2 promotes tumorigenesis of prostate cancer cells through up-regulation of EGF receptor (EGFR) signaling. Tumor growth of a prostate cancer cell line, DU145, was strongly decreased by STAP-2 knockdown. EGF-induced gene expression and phosphorylation of AKT, ERK, and STAT3 were significantly decreased in STAP-2-knockdown DU145 cells. Mechanistically, we found that STAP-2 interacted with EGFR and enhanced its stability by inhibiting c-CBL-mediated EGFR ubiquitination. Our results indicate that STAP-2 promotes prostate cancer progression via facilitating EGFR activation., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)

Chronic myeloid leukemia is a clonal disease characterized by the presence of the Philadelphia chromosome and its oncogenic product, BCR-ABL, which activates multiple pathways involved in cell survival, growth promotion, and disease progression. We previously reported that in murine hematopoietic Ba/F3 cells, signal transducing adaptor protein-2 (STAP-2) binds to BCR-ABL and up-regulates BCR-ABL phosphorylation, leading to enhanced activation of its downstream signaling molecules. The binding of STAP-2 to BCR-ABL also influenced the expression levels of chemokine receptors, such as CXCR4 and CCR7. For the induction of CCR7 expression, signals mediated by the MAPK/ERK pathway were critical in Ba/F3 cells expressing BCR-ABL and STAP-2. In addition, STAP-2 cooperated with BCR-ABL to induce the production of CCR7 ligands, CCL19 and CCL21. Our results demonstrate a contribution of CCR7 to STAP-2-dependent enhancement of BCR-ABL-mediated cell growth in Ba/F3 cells., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The promyelocytic leukemia protein PML acts as a tumor suppressor by forming transcription-regulatory complexes with a variety of repressor proteins. In the present study, we found that endogenous PML suppresses interleukin (IL)-6-induced gene expression as well as phosphorylation and transcriptional activation of STAT3 in hepatoma cells. We also found that PML-mediated suppression of IL-6-induced STAT3 activation by disrupting interactions between STAT3 and HDAC3. These results indicate that PML modulates IL-6-induced STAT3 activation and hepatoma cell growth by interacting with HDAC3., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Ectoparasitic arthropods are often vectors of rickettsiosis. We conducted a survey of ectoparasites on U.S. military facilities throughout Japan with the use of specimens submitted by pest control, public health, and veterinary personnel. Over 1,600 individual ectoparasites were collected. Fifteen species were identified, including several significant vectors of human diseases such as scrub typhus and rickettsial spotted fevers. These ectoparasites included Ctenocephalides felis , Haemaphysalis longicornis, Ixodes persulcatus , Leptotrombidium fuji, Leptotrombidium pallidum, and Rhipicephalus sanguineus . Rickettsial agents were detected by PCR and DNA sequencing. These included Bartonella henselae , Bartonella japonica, a novel Bartonella, Coxiella burnetii , an unnamed Coxiella, Ehrlichia canis , Orientia tsutsugamushi , Rickettsia typhi , and "Rickettsia Rf2125"/"Rickettsia cf1and5".

This report includes the distribution records of the Anopheles (Anopheles) Hyrcanus Group and associated species in Kyushu Island, Japan, based on our field collections from various localities of 4 prefectures (Fukuoka, Kumamoto, Nagasaki, Saga), primarily from 2002-2013. The status of common and potential mosquito vectors, particularly Anopheles species, in Japan are noted.

Mosquito collections were carried out in August 2002 and July 2003 in Japan. Anopheles sinensis of the Hyrcanus Group, Myzomyia Series of Anopheles, was the most common species encountered. The distribution and habitats of 5 Anopheles Hyrcanus Group species are included. Eight species belonging to Aedes, Culex, and Uranotaenia were found associated with An. sinensis in rice paddies and a variety of other larval habitats.

To investigate left ventricular wall motion asynchrony in patients with hypertensive heart disease, we measured regional myocardial velocity in hypertensive patients with left ventricular hypertrophy and in normotensive individuals using tissue Doppler imaging. The endocardial velocity and the myocardial velocity gradient were measured in the basal and mid segments of the septal and posterior walls. The dilating velocity of the left ventricular cavity were determined for the basal and mid ventricular segments of left ventricle. The peak myocardial velocity gradient was significantly lower in the hypertensive group than in the control group for all regions. The peak endocardial velocity during early diastole in the mid ventricular septum was significantly lower in the hypertensive group (Hypertensive vs Controls; 3.8 +/- 1.3 vs 5.1 +/1.6 cm/s, P < 0.05), whereas the peak endocardial velocity at the other three sites were similar in the two groups. The peak dilating velocity was significantly lower in the hypertensive group only in the mid portion of the left ventricle (Hypertensive vs Controls; 7.2 +/- 2.4 vs 9.8 +/- 1.3 s(-1), P < 0.005). These results suggest that there were regional wall motion abnormalities and nonuniformity during the early diastolic phase in the hypertensive hearts with left ventricular hypertrophy.

It has been recognized that arterial enlargement occurs in relation to the formation of atherosclerotic plaque. Previous studies on arterial remodeling have disregarded the role of mechanical deformation and have suggested that compensatory mechanisms occur to maintain arterial flow. We postulated that primary atherosclerotic enlargement and mechanical deformation are the predominant causes of the arterial remodeling. This hypothesis better explains the morphological changes without suggesting additional biological reactions.

The effects of changes in preload and chamber compliance on the transmitral flow pattern was investigated with a fluid dynamics model. A decrease in compliance led to a steepening of the deceleration slope and a shortening of the pressure half-time in both restricted and nonrestricted valves. An increase in the preload led to a longer pressure half-time but did not affect the slope in the restricted valve. However, increased preload led to a shorter pressure half-time and a steeper slope in the nonrestricted valve. A mathematically derived method to measure the net atrioventricular compliance (-[annular area] va/[pvt [flow deceleration rate]]), where va and vt are flow velocities at the mitral anulus and the mitral tip levels) was then proposed. The values obtained by this method showed a strong correlation with true values (r2 = 0.89). A possibility of noninvasive quantitative assessment of diastolic filling function was implied.

We measured Doppler echocardiographic diastolic parameter during infusion of low-dose dobutamine in 14 untreated hypertensive subjects and in 14 normal controls. Low-dose dobutamine accelerated left ventricular relaxation in normal controls but not in hypertensive subjects.

A 58-year-old woman with a prosthetic mitral valve and an anomalous single coronary artery received transesophageal echocardiography and was found to have a coronary artery--to--left atrium fistula. Because of its superior imaging quality, transesophageal color Doppler method is a useful diagnostic procedure.

Twenty-five patients with chronic aortic regurgitation (AR), and 12 control subjects were studied using Doppler echocardiography to investigate the effects of AR on transmitral flow. Peak early filling velocities at the levels of the mitral valve tips (E1) and annulus (E2) were measured, and the transmitral flow restriction index (delta E = (E1-E2)/E2) was obtained. Patients with AR were classified into 2 groups according to the ratio of the cross-sectional area of the regurgitant jet to that of the left ventricular outflow tract. Group I had the ratio less than 0.20, and group II had greater than or equal to 0.20. E2 in group II was lower than in control subjects, whereas E1 was not significantly different in any groups. delta E in group II was higher than in group I or in control subjects (p less than 0.05 and 0.01, respectively). delta E showed a significant correlation with the cross-sectional area ratio in all patients with AR (r = 0.70, p less than 0.01) and in group II (r = 0.82; p less than 0.01). Our data suggest that AR restricts early transmitral filling, and that delta E may indicate the increased driving pressure caused by flow restriction and is a useful hemodynamic index of AR.