Intestinal inflammations increase efflux of innate lymphoid cells from the intestinal mucosa to the mesenteric lymph nodes through lymph-collecting ducts.

INTRODUCTION: Innate lymphoid cells (ILCs) are abundant in the intestinal mucosa, forming boundaries externally. Herein, ILCs were directly obtained from intestinal lymph using a lymph fistula rat model and analyzed under physiological and pathological conditions. METHODS: Thoracic duct (TD) lymphocytes were collected by cannulation with/without preceded mesenteric lymphadenectomy, which were comparable to lymphocytes flowing through mesenteric lymphatic vessels (MLVs) or TD, respectively. The collected ILCs were classified according to gene transcription factors and analyzed by flow cytometry. The effect of IL-25 or indomethacin was studied. RESULTS: The proportion of total ILCs in the MLVs (MLV-ILCs) was significantly higher than that in TD (TD-ILCs, 0.01% vs. 0.003%, respectively). Physiologically, there were several significant differences in the MLV-ILCs compared with TD-ILCs, including the proportion of ILC2 (42.3% vs. 70.9%) and ILC3 (33.3% vs. 13.8%), and the proportion of α4-integrin-positive cells (36.8% vs. 0.3%). IL-25 significantly increased the proportion of MLV-ILC2 after 3 days. Indomethacin-induced intestinal injury increased the proportion of MLV-ILC3 in the early phase within 12 h. CONCLUSION: Intestinal ILCs were found to migrate through MLVs. The altered mobilization of MLV-ILCs after stimuli suggests that ILCs play an important role in regulating the immune responses at the secondary lymph nodes.

CD47 is an adverse prognostic factor and a therapeutic target in gastric cancer.

CD47 is an antiphagocytic molecule that acts via ligation to signal regulatory protein alpha on phagocytes; its enhanced expression and therapeutic targeting have recently been reported for several malignancies. However, CD47 expression in gastric cancer is not well documented. Immunohistochemical expression of CD47 in surgical specimens was investigated. Expression of CD47 and CD44, a known gastric cancer stem cell marker, were investigated in gastric cancer cell lines by flow cytometry. MKN45 and MKN74 gastric cancer cells were sorted by fluorescence-activated cell sorting according to CD44 and CD47 expression levels, and their in vitro proliferation, spheroid-forming capacity, and in vivo tumorigenicity were studied. In vitro phagocytosis of cancer cells by human macrophages in the presence of a CD47 blocking monoclonal antibody (B6H12) and the survival of immunodeficient mice intraperitoneally engrafted with MKN45 cells and B6H12 were compared to experiments using control antibodies. Immunohistochemistry of the clinical specimens indicated that CD47 was positive in 57 out of 115 cases, and its positivity was an independent adverse prognostic factor. Approximately 90% of the MKN45 and MKN74 cells expressed CD47 and CD44. CD47(hi) gastric cancer cells showed significantly higher proliferation and spheroid colony formation than CD47(lo) , and CD44(hi) CD47(hi) cells showed the highest proliferation in vitro and tumorigenicity in vivo. B6H12 significantly enhanced in vitro phagocytosis of cancer cells by human macrophages and prolonged the survival of intraperitoneal cancer dissemination in mice compared to control antibodies. In conclusion, CD47 is an adverse prognostic factor and promising therapeutic target in gastric cancer.

Role of caspase 8 as a determinant in chemosensitivity of p53-mutated head and neck squamous cell carcinoma cell lines.

We studied factors that control chemoresistance to 6 head and neck squamous cell carcinoma cell lines carrying p53 mutations. Cell lines were chosen, based on the presence of EGFR amplifications, the presence of H-ras mutations, and the absence of either. WST-1 viability assays showed that, in response to etoposide, Ca922 was most sensitive, HOC313 most resistant, and HSC6 and the others moderately sensitive. A similar tendency was shown by further analyses with cisplatin, 5-fluorouracil, LY294002, and combined treatment with LY294002 and TNF-related apoptosis-inducing ligand (TRAIL). Although both Ca922 and HOC313 had activating mutations upstream of Akt signaling, the constitutive phosphorylation of Akt at S473 was observed in chemosensitive Ca922, but not in chemoresistant HOC313, suggesting that constitutive Akt phosphorylation was not the primary determinant for chemoresistance in these cell lines. Further, by the combined treatment with LY294002 and TRAIL, apoptosis was induced in Ca922 and HSC6 but not in HOC313. Interestingly, caspase 8 was not detected in HOC313, while it was cleaved in the other 2 cell lines. Further, in Ca922 and HSC6 but not in HOC313, caspase 8 inhibitor restored loss of viability induced either with LY294002 and TRAIL or even with etoposide alone. These findings suggest that caspase 8 played an important role in chemoresistance against genotoxic drugs.

Tissue engineering of articular cartilage with autologous cultured adipose tissue-derived stromal cells using atelocollagen honeycomb-shaped scaffold with a membrane sealing in rabbits.

Adipose tissue derived stromal cells (ATSCs), which were isolated from adipose tissue of rabbit, have shown to possess multipotential, that is, they differentiate into osteoblasts and adipocytes in plate-culturing and into chondrocytes in an established aggregate culture using defined differentiation-inductive medium. The aim of this study was to evaluate the utility of ATSCs in tissue engineering procedures for repair of articular cartilage-defects using the atelocollagen honeycomb-shaped scaffold with a membrane sealing (ACHMS-scaffold). We intended to repair full-thickness articular cartilage defects in rabbit knees using autologously cultured ATSCs embedded in the ACHMS-scaffold. ATSCs were incubated within the ACHMS-scaffold to allow a high density and three-dimensional culture with control medium. An articular cartilage defect was created on the patellar groove of the femur, and the defect was filled with the ATSCs-containing ACHMS-scaffold, ACHMS-scaffold alone, or empty (control). Twelve weeks after the operation, the histological analyses showed that only the defects treated with the ATSCs-containing ACHMS-scaffold were filled with reparative hyaline cartilage, highly expressed Type II collagen. These results indicate that transplantation of autologous ATSCs-containing ACHMS-scaffold is effective in repairing articular cartilage defects.

Significance of angioscopic morphology for the estimation of macrophage infiltration and vascular physiology.

OBJECTIVE: The aim of the present study was to determine whether direct visual morphology can predict vascular physiology, macrophage infiltration and plaque architecture of atherosclerotic lesions. METHODS: Twenty male New Zealand white rabbits weighing 2.5-2.8 kg (mean, 2.7 +/- 0.2 kg) were used. We fed rabbits a 1% cholesterol diet after creating an injury in the middle abdominal aorta using a 2 Fr Fogarty balloon catheter. After 8 weeks, the morphology of plaque lesions was evaluated by angioscopy, and was classified as protruding or lining. Vascular endothelial function (VEF) was evaluated using a Doppler guidewire, and was expressed as ratio of basal to peak velocity (cm/s) in 30 s occlusion using a balloon catheter. Macrophages obtained from abdominal sections were stained with monoclonal antibody against rabbit macrophages (RAM-11). Density of total macrophage cell infiltration was quantified as absolute area of RAM-11 staining. RESULTS: Macrophage density and intima/media (I/M) ratio were significantly higher in the protruding group than in the lining group: macrophage density, 40 +/- 10 vs. 5 +/- 10%, p < 0.01; I/M ratio, 1.2 +/- 1.4 vs. 0.2 +/- 4, p < 0.05. Vascular flow reserve (VFR) was lower in the protruding group than in the lining group (1.8 +/- 0.7 vs. 2.2 +/- 0.5, p < 0.05). There was significant negative correlation between VEF and macrophage cell density (r = -0.593, p < 0.01), whereas there was no significant correlation between VEF and I/M ratio (r = -0.332, p = 0.330). CONCLUSION: The present findings suggest that protruding lesions revealed by angioscopy are rich in macrophages and have a higher I/M ratio and lower VFR.

Ramatroban, a thromboxane A2 receptor antagonist, prevents macrophage accumulation and neointimal formation after balloon arterial injury in cholesterol-fed rabbits.

Macrophage infiltration appears to play an important role in restenosis after arterial intervention. Monocyte chemoattractant protein-1 (MCP-1) is a major chemotactic factor for macrophages. We have previously shown that ramatroban, a thromboxane A(2) (TXA(2)) receptor antagonist, diminished the expression of MCP-1 in human vascular endothelial cells. The aim of this study was to evaluate whether, after balloon angioplasty of atherosclerotic arteries, ramatroban would reduce MCP-1 expression, macrophage accumulation, and neointimal formation. New Zealand white rabbits were fed a cholesterol-rich diet for 4 weeks, and the abdominal aorta of the rabbits were injured by a 2-French Fogarty catheter. They were randomized to receive 1 or 5 mg/kg daily of ramatroban (n = 7 or n = 8) or saline (n = 6). At 4 weeks after balloon angioplasty, the intimal hyperplasia and the macrophage-positive area in the intima by the ramatroban treatment was significantly reduced. Monocyte chemoattractant protein-1 gene expression in injured aortas of the ramatroban-treated group was significantly less evident than in the vehicle-treated group. Thromboxane A(2) receptor blockade by ramatroban for 4 weeks after balloon angioplasty in the atherosclerotic rabbits prevented macrophage infiltration through MCP-1 downregulation and neointimal formation.

A rapid speech synthesizing software on a PDA for Japanese with speech impairments.

We developed a Japanese-language, rapid synthesizing software application for use on a personal digital assistant. It has an unrestricted vocabulary and can synthesize words and sentences within 3 s. Eight hundred common sentences and words are preregistered. By touching the first character at the head of a preregistered sentence or word from an on-screen Kana (Japanese alphabet) chart, the user can select the sentence or word to be spoken. Characters on the Kana chart can also be input sequentially. Two Japanese subjects with speech impairments rated the device highly for its portability and quick response. Whereas communication previously had to be done by writing or sign language, it was easy for listeners with or without specialized training in communication with persons with speech impairments to understand the output from this device, making conversation easier which, in turn, improved the quality of life and social activity of these persons with speech impairments.

A novel ferromagnetic thermo-stent for plaque stabilization that self-regulates the temperature.

The purpose of this study is to investigate the vascular wall with a thermally self-regulating, cylindrical stent made of a low Curie temperature ferromagnetic alloy. Physiologic saline was circulated in the silicone model vessel implanted with the stent. The stent-temperature remained nearly constant for variable saline flows, saline temperatures, and magnetic flux densities. Stent implants of this type in human blood vessels could potentially enable thermotherapy and temperature determination without catheterization.

A network system of medical and welfare information service for the patients, their families, hospitals, local governments, and commercial companies in a medical service area.

A service information system using the Internet, which connected the various people who are related to medical treatment and nursing welfare, was constructed. An intractable neurological disease patient who lives in the Onga district, Fukuoka, Japan, and the people who are related to the service were chosen as test users in an experimental model. The communicated service information was divided into open-use data (electronic bulletin board, welfare service, medical care service, and link to private company service home page) and closed-use data (the individual patient's hysterics). The open data server was installed in an Internet service provider The open data could be accessed not only by the patient, but also by the family, information center, companies, hospitals, and nursing commodity store related to patient's nursing and medical treatment. Closed data server was installed in an information center (public health center). Only patient and information center staff can access the closed data. Patients should search and collect the service information of various medical and welfare services by themselves. Therefore, services prepared for the patient are difficult to know, and they cannot be sufficiently utilized. With the use of this information system, all usable service information became accessible, and patients could easily use it. The electronic bulletin board system (BBS) was used by patients for knowing each other or each others' family, and was used as a device for exchange of wisdom. Also, the questions for the specialist, such as doctor, dentist, teacher, physical therapist, care manager, welfare office staff member, and public health nurse, and the answers were shown on the BBS. By arranging data file, a reference of various patients in question and answer, which appeared in this BBS, was made as "advisory hints" and was added to the open data. The advisory hints became the new service information for the patients and their family. This BBS discovered the possibility of becoming an important information source for companies, hospital and, administration to know the requirements of patients and their families and the kind of services to be served. Although suppliers provide medical and welfare services for the patient, there is a tendency that the service information is sent by the suppliers at their own convenience. The information system in which various people participated was constructed in order to collect information for the patient, taking a patient-oriented approach. The result of the model test showed that this information system using Internet technology is a good system for both the service supplier and its receiver.

A practical procedure to prevent electromagnetic interference with electronic medical equipment.

Problems involving electromagnetic interference (EMI) with electronic medical equipment are well-documented. However, no systematic investigation of EMI has been done. We have systematically investigated the causes of EMI. The factors involved in EMI were determined as follows: 1) Electric-field intensity induced by invasive radio waves from outside a hospital. 2) Residual magnetic-flux density at welding points in a building. 3) Electric-field intensity induced by conveyance systems with a linear motor. 4) The shielding capacity of hospital walls. 5) The shielding capacity of commercial shields against a wide range frequency radio waves. 6) The immunity of electronic medical equipment. 7) EMI by cellular telephone and personal handy-phone system handsets. From the results of our investigation, we developed a following practical procedure to prevent EMI. 1) Measurement of electric-field intensity induced by invasive radio waves from outside the hospital and industrial systems in the hospital. 2) Measurement of residual magnetic-flux density at electric welding points of hospital buildings with steel frame structures. 3) Control of the electromagnetic environment by utilizing the shielding capacity of walls. 4) Measurement of the immunity of electronic medical equipment. And 5) Installation of electronic gate equipment at the building entrance to screen for handsets.