Extracellular Vesicles of Stem Cells to Prevent BRONJ.

Extracellular vesicles (EVs), several tens to hundreds of nanometers in size, are vesicles secreted by cells for intercellular communication. EVs released from mesenchymal stem cells (MSC-EVs) have the potential to treat multiple diseases. This study aimed to determine the effects of MSC-EVs on bisphosphonate-related osteonecrosis of the jaw (BRONJ), whose pathogenesis and treatment are not yet established. To this end, zoledronic acid (ZOL) was administered to bone marrow cells and fibroblasts in vitro. In vivo, a BRONJ model was produced by administering ZOL to rats and extracting teeth. Each MSC-EV-treated and nontreated group was compared histologically and molecularly. In vitro, the nontreated group showed an increased number of ß-galactosidase-positive cells and expression of senescence-associated genes p21, pRB and senescence-related inflammatory cytokines. Conversely, MSC-EV administration decreased the number of senescent cells and expression levels of p21, pRB and inflammatory cytokines. In vivo, in the nontreated group, the socket was partially uncovered by the oral epithelium, leaving an exposed bone. Conversely, in the MSC-EV-treated group, the socket was healed. Besides, in the nontreated group, ß-galactosidase-positive cells existed in the socket and colocalized with the CD90 and periostin-positive cells. However, there were few ß-galactosidase-positive cells in the MSC-EV-treated group. Furthermore, gene expression of stem cell markers Bmi1 and Hmga2 and the vascular endothelial marker VEGF was significantly increased in the MSC-EV-treated group, compared with that in the nontreated group. These results indicate that MSC-EVs prevent ZOL-induced senescence in stem cells, osteoblasts, and fibroblasts and reduce inflammatory cytokines. Furthermore, administration of MSC-EVs prevented senescence of cells involved in wound healing and the spread of chronic inflammation around senescent cells, thereby promoting angiogenesis and bone regeneration and preventing BRONJ.

Detection of centrosome amplification as a surrogate marker of dysfunction in the p53 pathway -p53 gene mutation or MDM2 overexpression.

Abstract In human and canine cancers, the inactivation of p53 protein as well as p53 gene mutation and MDM2 overexpression result in centrosome amplification that in turn contributes to chromosomal instability. To explore the usefulness of the detection of centrosome amplification as a surrogate marker of dysfunction in the p53 pathway, we systematically analysed centrosome amplification, p53 overexpression, p53 gene mutation and MDM2 overexpression in canine tumours. Centrosome amplification was detected in 16 of 51 (31%) naturally developing tumours in dogs. All the tumour specimens with aberrations in the p53 pathway, including p53 overexpression, p53 gene mutation or MDM2 overexpression, showed centrosome amplification, suggesting that the detection of centrosome amplification could serve as a preliminary surrogate marker of dysfunction in the p53 pathway.

Natural history of a patient with malignant nonfunctioning islet cell tumor associated with unresectable multiple liver metastases.

Islet cell tumors of the pancreas are uncommon. Approximately 15% of islet cell tumors are nonfunctioning and have a higher malignancy rate than their functioning counterparts. Though, because of the rarity of malignant nonfunctioning islet cell tumors, the natural history of a patient with this tumor has not been clearly defined. We describe a young patient with unresectable malignant nonfunctioning islet cell tumor associated with multiple liver metastases. He was treated with palliative therapies to improve his quality of life, but did not undergo surgical removal of tumors or systemic chemotherapy. He survived for 46 months since laparotomy for histological diagnosis. Our findings may represent the natural history of patients with unresectable malignant nonfunctioning islet cell tumor, and suggest that palliative therapy may contribute not only to the improvement of a patients' quality of life but also the prolongation of survival.

A small leiomyosarcoma of the pancreas treated by local excision.

BACKGROUND: Leiomyosarcoma of the pancreas is a very rare tumour; only 33 cases have been reported in the English literature. Because these tumours tend to be large and locally advanced, extended resection such as pancreatoduodenectomy or distal pancreatectomy has been advocated. CASE OUTLINE: A 25-year-old woman with left-sided back pain had a small low-density lesion (2 x 2 cm) in the pancreatic body on computed tomography (CT). The tumour was initially diagnosed as a pseudocyst and was managed conservatively. A repeat CT scan 10 months later showed an increase in the size of the lesion (3.5 x 3.5 cm) with contrast enhancement of a high-density area. The diagnosis was therefore changed to papillary-cystic neoplasm. At laparotomy, the tumour seemed to be arising from the mid-body of the pancreas with no invasion into surrounding organs. Local excision of the tumour was performed. Pathological examination revealed a leiomyosarcoma of the pancreas. DISCUSSION: Complete surgical resection offers the only potential chance of cure for patients with leiomyosarcoma of the pancreas.The lack of evidence of recurrence at three-and-a-half years suggests that the less invasive surgical procedure in the present case has been curative. Careful and intraoperative exploration was important to determine the best surgical procedure.

Mother's working status and nutritional status of children under the age of 5 in urban low-income community, Surabaya, Indonesia.

A cross-sectional anthropometrics survey was carried out in a low-income community of Surabaya, Indonesia, to examine the association between mother's employment and the child's nutritional status for identifying a group at risk of having malnourished children. Subjects were 274 children under the age of 5. The children of non-working mothers had significantly higher height-for-age z-score (HAZ) (p < 0.05) than those of working mothers. When mother's work was divided into 'formal' and 'informal', HAZ and weight-for-age z-score (WAZ) of children of the informal worker's group were significantly lower than those of the non-working mother and the formal worker's group (p < 0.05). Mother's education and income of the formal worker's group were significantly higher than those of the informal worker's group (p < 0.01). Our study identified a group at risk of malnutrition, i.e., children of mothers working in informal sectors. The programs to provide childcare for working mothers should target this particular group of poor households.

The effect of additional autopolyploidization in a slow growing cellulase hyperproducer of Trichoderma.

M14-2 is a cellulase hyperproducer derived from Trichderma reesei QM 6a, but with a growth rate lower than that of the original strain. When M14-2 was autopolyploidized followed by haploidization and selection, the strain with both a higher cellulase productivity per mycelia and a higher growth rate could be obtained as M14-2B. This strain seemed to be constructed using gene sources amplified by additional autopolyploidization.

Successive construction of cellulase hyperproducers of Trichoderma using hyperpolyploids.

When the swollen conidia of Trichoderma reesei QM 6a are treated with 0.1% (w/v) colchicine solution, huge autopolyploid nuclei can be formed in those swollen conidia. When a mycelial mat derived from such a conidum is treated with a haploidizing reagent, benomyl, many fan-shaped sectors are produced from the colony, and cellulase hyperproducers are selected from conidia on the colony. When colchicine and benomyl treatments are repeated on cellulase hyperproducers, new hyperproducers can be constructed successively and systematically. Moreover, when conidia derived from autopolyploids are treated with ethylmethanesulfonate solution, another type of cellulase hyperproducers (polyploids) can be obtained.

Cellulase hyperproducers constructed from polyploids of Lentinus edodes.

A mycelial mat of Lentinus edodes was treated with 0.01% (w/v) colchicine solution for 240 h at 26 degrees C and autopolyploidization occurred. The mycelia were treated subsequently with the haploidizing reagent, benomyl, and fanshaped sectors were produced from colonies. Among such sectors, cellulase hyperproducers could be selected. The cellulase productivity of the hyperproducer, L1, did not decrease through five generations.

Construction of cellulase hyperproducing strains derived from polyploids of Trichoderma reesei.

The mycelial mat of Trichoderma reesei strain QM 6a was treated with 0.1% (w/v) colchicine solution for 14 days and designated M14. The cellulase productivity of strain M14 was not much higher than that of the original strain. When conidia of M14 were treated with ethylmethane sulphonate (EMS) solution, the cellulase hyperproducers, M14-1 and M14-2, were isolated using a selection medium containing Avicel. The DNA content of M14-1 and M14-2 was higher than that of the original strain. Cellulase productivity per mycelium of these strains increased and was higher than that of the original strain. The cellulase productivity did not change through ten generations when these strains were cultivated successively on a medium containing Avicel. It was concluded that cellulase hyperproducers, whose cellulase productivity per mycelium increased, could be obtained when the conidia of strain M14 were treated with EMS.

Endoscopic findings in transplanted allo-intestine of rats after discontinuance of immunosuppressive agent.

In clinical practice, graft rejection in small-bowel transplantation should be diagnosed before irreversible condition of the graft. We have already reported the usefulness of endoscopic examination for the early detection of acute rejection in a rat model. Here we evaluated rejection after discontinuance of methyl-deoxyspergualin by endoscopy. Heterotopic small-bowel transplantation was performed by the cuff method from a DA to a LEW rat. Endoscopic and histological examinations were performed through the stomas. Two-week administration of methyl-deoxyspergualin significantly prolonged graft survival. Graft rejection after discontinuance of the agent occurred much more slowly than rejection without the immunosuppressive drug. Erosive mucosal changes were endoscopically observed in the early phase of rejection in rats that did not receive the immunosuppressant. However, endoscopic findings after discontinuance of methyl-deoxyspergualin indicated edematous changes and thickening of the wall without erosion, and, histologically, the grafted intestine showed slowly-progressing rejection with flattened villi. If we pay attention to edematous changes and hardening of intestinal wall, and take selective biopsies, endoscopic examination may improve the early diagnosis of slowly progressive rejection in the clinical setting.

Effect of fibrin glue on small and large bowel anastomoses in the rat.

Fibrin glue has been used as a hemostatic and adhesive agent for many years, but its efficacy in digestive surgery is still controversial. In this study, we evaluated the effect of fibrin glue on the healing of the small and large bowel anastomotic portions by measurement of the anastomotic bursting pressure. Further, the efficacy of fibrin glue in preventing intra-abdominal adhesion formation was assessed. Our results indicated that fibrin glue enhanced the resistance towards an elevation of intraluminal pressure in small bowel, but not in large bowel. Concomitantly, the glue did not significantly attenuate postoperative adhesion formation. A sealing action of fibrin glue might protect against bleeding and microleakage, resulting in beneficial effects on small bowel anastomosis.

Hormone-cytokine response. Pneumoperitoneum vs abdominal wall-lifting in laparoscopic cholecystectomy.

BACKGROUND: Changes in blood hormone and cytokine were investigated in patients who underwent laparoscopic cholecystectomy via insufflation (CO2 group) vs those who had abdominal wall-lifting (Air group). METHODS: Seventeen female patients with cholecystolithiasis were randomly divided into two groups. Peripheral blood samples were obtained during perioperative period, and plasma hormone levels (ACTH, cortisol) and serum cytokine levels (TNFalpha, IL-1beta, IL-6, IL-10) were measured. RESULTS: The number of circulating lymphocytes significantly decreased at 1 h after surgery in both groups, but the decrease in the CO2 group was significantly smaller than that in the Air group. There was no significant difference in hormone elevation between groups. Serum concentrations of IL-6 and IL-10 in the Air group were significantly higher than in the CO2 group. CONCLUSIONS: CO2 insufflation may reduce cytokine production in laparoscopic cholecystectomy.

[Awareness of health and welfare planning for elderly and methods to increase awareness: based on social marketing].

To clarify the socio-psychological factors affecting awareness of health and welfare planning for the elderly, a survey of community residents was performed in the Tokyo metropolitan area. The results obtained were as follows: 1. There were three factors which affected awareness concern about health and welfare services for the elderly as a social issue: concern about the local community; and concern about the local government. 2. Two other factors which did not affect the level of awareness were: anxiety about the health and welfare services for the elderly as a personal issue; and having personal experience of nursing care. From these results, a method to increase awareness was studied based on social marketing methods. Two primary target groups for increasing awareness were identified. One target was people who have the socio-psychological factors described above. Another target was people who have the potential need for health and welfare services but who were not aware of it. That is, they have relatively high anxiety about health and welfare services for the elderly as a personal issue and have personally experienced nursing care. The method of approach for these targets were studied. For the first group, the amount of the information available seemed to be important because these persons are ready to recognize the need for planning. Therefore, efficient information channels should be selected. For the second group, approaches that generate greater consumer participation by presenting this as an efficient method for solution of the problem should be adopted.

Segmental small bowel transplantation in the rat: comparison of lipid absorption between jejunal and ileal grafts.

BACKGROUND: From the immunological point of view, it is very important to evaluate the efficacy of segmental small bowel transplantation and to determine which part of the intestine, jejunum, or ileum should be used. In the present study, we investigated the absorptive function of the transplanted jejunum and ileum in the rat. METHODS: Syngeneic segmental small bowel transplantation (jejunal or ileal grafting) was performed. After surgery, body weight and fecal fat excretions were measured. In addition, bile acid concentration in bile juice was measured, and the response of both serum lipase and bile flow rate after oleic acid stimulation was evaluated. Recanalization of the lymphatic vessels was investigated by lymphangiography. RESULTS: There was no significant difference in body weight change between normal controls, jejunum-transplanted rats, and ileum-transplanted rats. In short gut rats, however, body weight was significantly impaired. Fecal fat excretion in short gut rats was the highest in the four groups, and it was significantly lower in ileal grafts than in jejunal grafts. Ileum-transplanted rats also showed a significantly higher bile acid concentration in bile juice than jejunum-transplanted rats. After oleic acid stimulation, serum lipase responded better in ileal transplants than in jejunal transplants, but the bile flow rate did not show significant change in either group. Recanalization of the lymphatic vessels was established on the 28th postoperative day. CONCLUSIONS: These results clearly show that ileal transplantation is more conducive to lipid absorption than jejunal transplantation.

Evidence that the elevation of soluble MHC class I antigens in the serum precedes the onset of graft-versus-host disease and is correlated with the severity of the disease in rats.

We examined the changes in the levels of soluble major histocompatibility antigen complex (MHC) class I antigens in the serum under a lethal or nonlethal state of graft-versus-host-disease (GVHD) induced by injecting various doses of PVG rat splenic lymphocytes into (DA x PVG)F1 rats. All rats receiving 4 x 10(8) lymphocytes (lethal dose) died on day 20-36 showing typical features of GVHD, while the injection of 4 x 10(7) cells (nonlethal dose) induced no sign of GVHD. When rats were inoculated with a nonlethal dose of lymphocytes prior to the injection of a lethal dose, all rats survived with or without showing transient GVHD. Preceding the onset of GVHD the levels of soluble class I antigens increased significantly to 1094 +/- 487 ng/ml (mean +/- SD, n = 4) from 3 days after the injection of a lethal dose to the time of death, whilst the levels in the nonlethal dose group remained unchanged. Rats with transient GVHD in the preinoculated group showed the increase of soluble class I antigens to the same extent as rats with lethal GVHD, suggesting that GVHD was systemically ongoing. The levels of soluble class I antigens also correlated with the severity of GVHD as judged by daily observation and histological studies. Rats receiving a lethal dose showed destructive alteration of spleen structure and cellular infiltration in the portal area of the liver before the animals started to show signs of GVHD, whereas rats in the nonlethal dose group exhibited no marked change. These data suggest the possibility of serum soluble class I antigens being not only a diagnostic but also a prognostic marker for GVHD.

Fulminant second-set allograft rejection and endoscopic findings following small bowel transplantation in the rat.

In the presensitized recipient who has been exposed to donor antigens, second-set rejection takes the form of severe hyperacute graft rejection. Second-set allograft rejection was studied following small bowel transplantation in the rat. Heterotopic intestinal grafting was performed from DA (RT1a) donors to PVG (RT1c) recipients 4 weeks after DA skin sensitization. The endoscopic images and histological specimens were compared with those of syngeneic and first-set rejected grafts. Endoscopically, diffuse erosions of the graft were detected from day 1. Mucosal necrosis progressed rapidly, and was accompanied by massive bleeding on days 3-5. These findings were similar to the course of severe necrotizing hemorrhagic enteritis. Histologically, interstitial edema and hemorrhage with massive infiltrations of neutrophils were manifested from day 1. Mesenteric vessels were completely occluded by thrombi on days 3-5. The grafted intestine had became totally necrotic by day 5. Microscopic findings strongly suggested that destructive graft necrosis was due to vascular damage caused by humoral factors. All the presensitized rats (n = 11) died showing systemic septic signs by day 11 after small bowel transplantation. We conclude that lethal hyperacute rejection occurred in presensitized recipients, even when the graft was transplanted heterotopically. Endoscopic evaluation is beneficial for the early diagnosis of graft rejection. Immediate graft removal should be mandatory as a rescue treatment in second-set rejection of the small intestine.

Fungal metabolites. XIX. Structural elucidation of channel-forming peptides, trichorovins-I-XIV, from the fungus Trichoderma viride.

Trichorovins (TV)-I-XIV are new antibiotic peptides obtained from conidia of the fungus Trichoderma viride. The peptide mixture of TVs was repeatedly fractionated by preparative HPLC until individual TVs showed a single peak on their analytical HPLC chromatograms. Nevertheless, FAB-MS or NMR indicated that each of TVs-I-XIV was composed of at least two components. We attempted to elucidate their structures within the fractions by electrospray ionization (ESI)-MS, FAB-MS, FAB-MS/MS and NMR. TVs generally have molecular weights of approximately 1100-1200 Da, and are characterized by an acetylated N-terminus, the presence of an aminoalcohol, e.g. leucinol, isoleucinol or valinol, at the C-terminus, and eleven residues including three alpha-aminoisobutyric acids in the molecule. Thus, it was determined that TVs belong to the class of peptaibols.