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Long-term in vivo observation in large animal model is critical for evaluating the potential of small diameter tissue engineering vascular graft (SDTEVG) in clinical application, but is rarely reported. In this study, a SDTEVG is fabricated by the electrospinning of poly(ε-caprolactone) and subsequent heparin modification. SDTEVG is implanted into canine's abdominal aorta for 511 days in order to investigate its clinical feasibility. An active and robust remodeling process was characterized by a confluent endothelium, macrophage infiltrate, extracellular matrix deposition and remodeling on the explanted graft. The immunohistochemical and immunofluorescence analysis further exhibit the regeneration of endothelium and smooth muscle layer on tunica intima and tunica media, respectively. Thus, long-term follow-up reveals viable neovessel formation beyond graft degradation. Furthermore, the von Kossa staining exhibits no occurrence of calcification. However, although no TEVG failure or rupture happens during the follow-up, the aneurysm is found by both Doppler ultrasonic and gross observation. Consequently, as-prepared TEVG shows promising potential in vascular tissue engineering if it can be appropriately strengthened to prevent the occurrence of aneurysm.

To create anti-adhesive materials to be more effective and safer, we developed a thermally cross-linked gelatin film that showed superior anti-adhesive effects with excellent peritoneal regeneration. However, it may act as a convenient scaffold for tumor cell growth, thereby accelerating peritoneal dissemination when used in surgery for abdominal tumors. In this study, we tried to clarify this issue using mouse carcinomatous peritonitis models. First, we examined the in vitro tumor cell growth of mouse B16 melanoma or Colon26 cells on the gelatin film or the conventional hyarulonate/carboxymethylcellulose film. Tumor cell growth on each film was significantly lower than that of the control (no film). Next, we conducted the following in vivo experiments: After the parietal peritoneum was partially removed and covered with each film or without any film, mice were inoculated intraperitoneally with B16 melanoma or Colon26/Nluc cells expressing NanoLuc luciferase gene. At 7 days after the operation, we measured the weight of B16 melanoma tumors or the NanoLuc activity of Colon26/Nluc cells using in vivo imaging at the injured sites. There were no significant differences in the weight of the tumors and the NanoLuc activity among the three groups. We also observed the survival time of mice receiving the same operation and treatments. There was no significant difference in the survival time among the three groups. These results suggest that the gelatin film will likely not accelerate peritoneal dissemination as a convenient scaffold for tumor cell growth when used in surgery for abdominal tumors. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2122-2130, 2018.

Aim Adhesion after pelvic surgery causes infertility, ectopic pregnancy, and ileus or abdominal pain. The materials currently available for clinical use are insufficient. The purpose of this study was to develop an anti-adhesive material that overcomes the limitations of conventional anti-adhesive agents. Methods The adhesion prevention effects of three methods – a two-layered sheet composed of gelatin film and gelatin sponge, Seprafilm and INTERCEED – were evaluated in 37 dogs. Anti-adhesive effects were investigated macroscopically and microscopically in a cauterized uterus adhesion model. Cell growth on the materials in vitro using human peritoneal mesothelial cells, fibroblasts and uterine smooth muscle cells were also evaluated. Results The two-layered gelatin sheet had significantly superior anti-adhesive effects compared to the conventional materials (Seprafilm and INTERCEED). A single-cell layer of mature mesothelium formed three weeks after surgery in the gelatin group. Peritoneum regeneration in the Seprafilm and INTERCEED groups was delayed and incomplete in the early phase. Little inflammation around the materials occurred and cell growth was significantly proliferated with the gelatin sheet. Conclusion The anti-adhesive effects of a two-layered gelatin sheet were superior to conventional agents in a cauterized canine uterus model, demonstrating early regeneration of the peritoneum, little inflammation and material endurance. The newly developed two-layered gelatin sheet is a useful option as an anti-adhesive agent for deeply injured and hemorrhagic sites.

Postoperative air leaks remain a major cause of morbidity after lung resection. This study evaluated the effect of a combination of polyglycolic acid (PGA) sheet and alginate gel on pulmonary air leaks in rats. Four pulmonary sealing materials were evaluated in lung injury: fibrin glue, combination of PGA sheet and fibrin glue, alginate gel, and combination of PGA sheet and alginate gel. With the airway pressure maintained at 20 cmH2O, a 2 mm deep puncture wound was created on the lung surface using a needle. Lowering the airway pressure to 5 cmH2O, each sealing material was applied. The lowest airway pressure that broke the seal was measured. The seal-breaking pressure in each experimental group was fibrin, 10.4±6.8 cmH2O; PGA + fibrin, 13.5±6.5 cmH2O; alginate gel, 10.3±4.9 cmH2O; and PGA + alginate, 35.8±11.9 cmH2O, respectively. The seal-breaking pressure was significantly greater in the PGA + alginate gel group than in the other groups (p<0.01). There were no significant differences among the other three groups. Alginate gel combined with a PGA sheet is a promising alternative to fibrin glue as a safe and low-cost material for air leak prevention in pulmonary surgery.

大動脈を再生するため、多層の管状構造の新規の血管再生足場を開発した。最内層はエレクトロスピニング法で作成したラクチド/カプロラクトン・コポリマー布のチューブである。メルトブロウ法あるいは編んで作成したラクチド/カプロラクトン・コポリマー及びポリラクチド布のロールが最内層の上に巻いてある。吻合部の外層はメルトブロウ法で作成したポリグリコール酸布で覆われている。この多層性の血管用足場をイヌの大動脈に移植した。経時的な組織再生と移植片の分解の詳細をラットとイヌにおいて観察した。その結果、10月後には本来の大動脈と殆ど同じ構造の大動脈が再生された。最初はそれ自身はポリマーの布であった外側ポリマー布層は、移植後早期に、自己組織化した層に置き換わった。この外側層は、最内層の補強材の働きをしていると考えられた。最内層のチューブの布は本来の構造を保持しつつ徐々に分解された。結論として、この新規の血管再生足場は、イヌの大動脈の構造と殆ど同じ構造の大動脈を再生することが可能である。, To regenerate the aorta we devised a new vascular scaffold with multilayered tube structure. The innermost layer was a fabric tube made of poly (lactide/caprolactone) produced by electro-spinning method. Fabric roles of poly (lactide/caprolactone) and/or poly (lactide) produced by melt-blow or knit method were overlying the innermost layer. An outer layer of anastomotic site was covered with a role of poly (glycolic acid) fabric produced by melt-blow method. The multilayered vascular scaffold was implanted in the native abdominal aorta of dog. Chronological observation of tissue regeneration and graft degradation were also conducted in rats and dogs for detail analyses. The aorta was regenerated in 10 months in almost similar structure to the native aorta. In the early phase after implantation, the outer polymer layers, initially polymer fabrics themselves, were replaced by self-organizing membranes. The outer layers were considered to work for reinforcement of the innermost layer. The fabrics of innermost tube maintained the original structure and degraded slowly. In conclusion the new vascular scaffold can regenerate the canine aorta with almost similar structure to the native aorta.

To create anti-adhesive materials to be more effective and safer, we developed a thermally cross-linked gelatin film that showed superior anti-adhesive effects with excellent peritoneal regeneration. However, it may act as a convenient scaffold for tumor cell growth, thereby accelerating peritoneal dissemination when used in surgery for abdominal tumors. In this study, we tried to clarify this issue using mouse carcinomatous peritonitis models. First, we examined the in vitro tumor cell growth of mouse B16 melanoma or Colon26 cells on the gelatin film or the conventional hyarulonate/carboxymethylcellulose film. Tumor cell growth on each film was significantly lower than that of the control (no film). Next, we conducted the following in vivo experiments: After the parietal peritoneum was partially removed and covered with each film or without any film, mice were inoculated intraperitoneally with B16 melanoma or Colon26/Nluc cells expressing NanoLuc luciferase gene. At 7 days after the operation, we measured the weight of B16 melanoma tumors or the NanoLuc activity of Colon26/Nluc cells using in vivo imaging at the injured sites. There were no significant differences in the weight of the tumors and the NanoLuc activity among the three groups. We also observed the survival time of mice receiving the same operation and treatments. There was no significant difference in the survival time among the three groups. These results suggest that the gelatin film will likely not accelerate peritoneal dissemination as a convenient scaffold for tumor cell growth when used in surgery for abdominal tumors. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2122-2130, 2018.

Background and study aims: Recently, the shielding method with polyglycolic acid (PGA) fabric and fibrin glue (P-F method) has been reported to prevent serious complications after endoscopic submucosal dissection (ESD). However, the effectiveness and mechanism to prevent complications by this method remain unclear and the corresponding basic research has not been fully conducted. Methods and results: We examined the effectiveness and mechanism of the P-F method, using a surgical ESD model of canine stomach and in vitro experiments. In the model experiment treated by P-F method or no treatment (control), ulcer perforation or penetration occurred only in the control group, but not in the P-F group. Microscopically, the P-F group showed less damages of the ulcer lesion than that of the control group, showing thicker granulation tissues including PGA fibers on the third day and excellent mucosal regeneration on the fourteenth day. In vitro culture experiments showed that fibroblasts proliferated at a significantly higher rate on PGA than on fibrin or a complex thereof. However, under hydrochloric acid treatment, fibroblasts were protected by fibrin, followed by the complex of both, and PGA. Conclusion: The P-F method exhibited a protective effect against gastric juice by fibrin glue to reduce tissue damages and a scaffold function of PGA fabric to induce better granulation formation at the earlier phase, resulting in excellent long-term tissue repair, on ulcer lesion following ESD, although the results were based on basic experiments.

Total knee arthroplasty (cemented) using a ceramic Yokohama Medical Ceramic Knee (YMCK) (Kyocera Corp; Kyoto, Japan) prosthesis was evaluated in 90 knees of 64 patients with severe rheumatoid arthritis, with a mean age of 59 ± 11 years. The mean follow-up period was 56 ± 20 months (range, 27 to 97 months). Larsen's grading of rheumatoid arthritis revealed grade 2 in 3, grade 3 in 36, grade 4 in 36, and grade 5 in 15 of 90 knees. Of 64 patients, 9 were judged to have class 1, 30 were judged to have class 2, and 25 were judged to have class 3 functional disability. The American Knee Society knee score and function score improved from 41 ± 16 and 29 ± 22 preoperatively to 83 ± 14 and 50 ± 29 at the final follow-up (P

The purpose of the study reported in this article was to investigate effects of synthetic lipid A on the expression of adhesion molecule CD44 on bovine articular chondrocytes. Full-thickness bovine articular cartilage was dissected from the carpometacarpal joints of 24 cows. Cartilage pieces were enzymatically digested to liberate chondrocytes. The chondrocytes were incubated in the presence of synthetic lipid A in suspension culture. Cell characteristics and binding of monoclonal antihuman CD44 antibodies were assessed with a flow cytometer. The expression of CD44 mRNA in chondrocytes was detected by reverse transcription-polymerase chain reaction (RT-PCR) technique. PCR products were quantified with a charge-coupled device image sensor. The percentage of CD44-positive chondrocytes was 42.2% ± 12.0%, 51.7% ± 6.8%, and 51.1% ± 5.0%, in the presence of lipid A at 0.25 μg/ml, 2.5 μg/ml, and 25 μg/ml, respectively, whereas it was 39.2% ± 8.9% in the absence of lipid A. In flow cytometry, two subpopulations of chondrocytes were found in each of five separate experiments, one with smaller number of forward scatter (FS) and the other with larger number of FS. The percentage of CD44-positive cells was 24.8% ± 8.5% in the subpopulation with smaller number of FS and 31.9% ± 6.4% in the subpopulation with larger number of FS at time 24 h after incubation. The bacterial component, lipid A, upregulated expression of CD44 on articular chondrocytes.

To study the effect of synthetic lipid A on the expression of the adhesion molecule CD44 in synovium and cartilage, synthetic lipid A (10 μg/ml, 0.5 ml) diluted in 0.025% triethylamine (TEA) was injected into the left knee of male Wistar rats. The equivalent volume of 0.025% TEA was injected simultaneously into the right knee of the same rat as a control. The numbers of infiltrating neutrophils and mononuclear cells in the synovium were counted, and the expression of CD44 was detected immunohistochemically. Infiltration of neutrophils in the synovium of the knee reached a maximum at 12 h after intraarticular injection of lipid A (78.4 ± 5.6 cells/400× field), and had subsided at 7 days after injection (2.1 ± 0.6). CD44 expression in the lining cells of the synovium was detected 24 h after injection (2.5 ± 0.3 cell layers), and it lasted 7 days after injection (2.6 ± 0.4 cell layers). CD44 expression in cartilage started at 24 h (69.0% ± 4.6% positive area of specimen) and lasted 7 days after injection (27.3% ± 3.3%). Intraarticular injection of lipid A was proven to induce acute arthritis and CD44 expression in the synovial lining layers and articular cartilage.

This study was designed to determine the effect of lipid microspheres containing prostaglandin E 1 (lipo PGE 1 ) on cis -diamminedichloroplatinum (CDDP) accumulation in primary and lung metastatic lesions. Sixty mice were divided into four groups, depending on whether or not an intra-foot-pad injection of Dunn osteosarcoma cells had been administered and on whether or not an intraperitoneal injection of lipo PGE 1 had been administered. CDDP was injected intraperitoneally into all the mice 6 weeks after the inoculation. Tumor colonies of spontaneous metastases in the left lung were found in 21 out of 30 tumor-inoculated mice at autopsy. Tissue platinum concentrations in the lungs with metastatic colonies and in the foot-pad tumors were significantly higher in the lipo PGE 1 -administered mice than in those without treatment. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) assay showed marked localization of dying cells in the lung metastatic lesions of the lipo PGE 1 -administered mice. The results of this study showed that pretreatment with lipo PGE 1 may augment the anti-tumor effects of CDDP at the tumor site.

A 53-year-old man recieved left-upper lobectomy for a suspecion of lung cancer associated with bullae, which was indicated by a rapid enlargement of abnormal shadow in the lung. Postoperative histologic and immunohistochemical examinations could first offer definite diagnosis of primary malignant fibrous histiocytoma of the lung, but pre-and intraoperatively we had difficulties in judging whether the tumor was benign or malignant. Malignant fibrous histiocytoma primary arising in the lung is so rare that only approximately 30 cases have been reported in the foreign and domestic literature. This extreme difficulty in diagnosing the disease might emphasize the importance of aggressive surgical treatment leading to early diagnosis and treatment. Primary malignant fibrous histiocytoma of the lung with bullous lesion has never been reported in the literature.

In the present study, the adjuvant effect of zymosan on human immunodeficiency virus type-1 (HIV-1)-specific DNA vaccine and the mechanism of this enhancement were studied in a murine model. We coinoculated zymosan with our candidate HIV-1 specific DNA vaccine (pCMV160IIIB) into skeletal muscles of BALB/c mice. Higher levels of both humoral immune response and HIV-specific delayed-type hypersensitivity (DTH) response were observed when zymosan was coinoculated with pCMV160IIIB compared with that obtained using pCMV160IIIB alone. HIV-specific cytotoxic T lymphocyte (CTL) activity was also enhanced. This enhancing activity was suppressed when coinoculated to the fifth complement (C5)-deficient DDD and AKR mice. The enhanced activity was also suppressed when anti-C3 antibody was inoculated to mice intramuscularly. There was significant induction of immunoglobulin G2a (IgG2a) and interferon-gamma (IFN-gamma) in pCMV160IIIB vaccine with zymosan. These results suggest that zymosan-mediated DNA vaccination enhances helper T cell (Th) 1-mediated immunity. The effect is suggested to be based on the consequences of its recruitment and activation of macrophages, dendritic cells or antigen-presenting cells (APC) through complement activation, especially through the alternative pathway. Taken together, these results suggest that zymosan can be an effective immunological adjuvant in DNA vaccination against HIV-1.

A porous hydroxyapatite (highly purified synthetic Ca10(PO4)6(OH)2) wedge was inserted into the tibia in 10 knees of 7 patients with osteoarthritis of the knee who underwent high tibial osteotomy. The interface of this HA wedge with bone was histologically examined in undecalcified specimens obtained at the time of hardware removal in all 10 knees, and total incorporation of the HA into bone was observed, with no inflammatory reaction. The proportion of pores that were filled with regenerated bone within 300 microm from the interface was 71.8 +/- 10.1% (mean +/- S.D.), and it was positively correlated with the time from implantation to biopsy. Radiodensity of the HA wedge was measured during the follow-up period, and it did not change significantly within 36 months after osteotomy. Our study concluded that bone ingrowth into the HA block slowly progressed and was correlated with the passage of time.

We reviewed 11 knees in ten patients at a mean follow-up of 23 years after a modified Loeffler's medial capsulotomy for osteoarthritis of the knee. The mean age was 59+/-8 years at surgery. The mean knee score (Hospital for Special Surgery) was 59 points before surgery and 79 points at follow-up. The long-term results of this operation were thus maintained over a prolonged period. This is a relatively simple procedure, which may be performed in the elderly, with satisfactory long-term results.

The promoting effects of matrix metalloproteinase-2 (MMP-2) on lung metastasis of human fibrosarcoma cells (HT1080) were studied using nude mice. The fourth generation of HT1080 was established by consecutive clonal selection of metastatic lung nodules formed by intravenous transplantation. MMP-2 and MMP-9 in the culture supernatants of the first and fourth generation cells were analyzed by gelatin zymography and Western blotting, and quantified by scanning densitometry. In gelatin zymograms, mean ratios of values for the 59-kDa band (the active form of MMP-2) to those for the 72-kDa band (the inactive form of MMP-2) for optical density; area, and volume measured by densitometry were 1.44 +/- 0.12, 0.93 +/- 0.05, and 1.27 +/- 0.20, respectively, in the culture supernatant of fourth generation cells isolated from metastatic lung nodules. These values were significantly (P0.01) higher than those of first generation cells (0.70 +/- 0.04, 0.48 +/- 0.01, and 0.57 +/- 0.42). Three weeks after intravenous transplantation of HT1080 cells into nude mice, the incidence of lung metastasis and mean number and diameter of metastatic nodules formed by injection of first generation cells were 20% (2 of 10 mice), 2.9 +/- 0.2 and 2.0 +/- 0.2 mm, respectively; while they were 100%, 99.8 +/- 7.2 and 4.3 +/- 0.3 mm following injection of fourth generation cells. These findings suggest that the active MMP-2 produced by human fibrosarcoma cells is important for the cells to form lung metastatic lesions in nude mice.

The anti-tumor effects of hypoosmotic solution of MTX in distilled water (DW) on Dunn osteosarcoma were evaluated in mouse air pouches. Dunn osteosarcoma cell suspension (1 x 10[5] cells in 0.1 ml of medium) was inoculated into the mouse subcutaneous air pouch that had formed 7 days after the initial injection of air. Two hours after the inoculation of tumor cells, 5 ml of various concentrations of MTX (from 0 to 1 x 10[-3] M) dissolved in DW or PBS were injected into the air pouch. Five minutes later, the entire solution in the air pouch was aspirated. The mice were sacrificed 3 weeks after the inoculation of tumor cells and the air-pouch tissue was transected in the coronal plane with the largest area of tumor mass. The sections were stained with HE and the area was measured with the NIH Image program. The largest area of tumor mass in the air pouch treated with 1 x 10(-3) M of MTX in DW was 11.8+/-3.4 mm2 (N = 5), which was significantly (P0.005) smaller than that in PBS (51.7+/-8.3 mm2). These findings suggested that hypoosmotic solution in DW might augment the anti-tumor effect of MTX on sarcoma cells.

Objective. Previous studies suggested that it was possible to characterize the intact surface of articular cartilage by probing it with antibodies against matrix macromolecules. The present studies were undertaken to investigate type II collagen (CII) epitope availability on the intact surface of articular cartilage. Methods. Normal bovine, rabbit, and human cartilage specimens were used to measure binding of anti-CII antibodies to the articular and cut surfaces of cartilage. Antisera were raised against the material obtained after brief extraction of the cartilage surface with 4M guanidine solution. Results. Anti-CII did not bind to the intact surface of rabbit articular cartilage when compared with control rat sera, but did bind significantly to the cut surface. With normal human articular cartilage, the cut surfaces bound approximately 4 times as much anti-CII antibody as the intact articular surfaces. These findings suggested that the CII epitopes are normally protected by the superficial cartilage layer. In experiments carried out to characterize this layer, binding of anti-CII antibodies was unchanged after exhaustive washing of bovine or rabbit cartilage with phosphate buffered saline or 1M NaCl solution, whereas it was significantly increased after brief exposure to 4M guanidine solution or after incubation with neutrophil elastase. No restoration of the protection of CII epitopes in guanidine-treated cartilage could be achieved by incubation with undiluted normal bovine synovial fluid; however, 8-day culture of elastase-treated cartilage explants resulted in partial restoration of protection of the CII epitopes. Rat and rabbit antisera raised against the cartilage surface material extracted with 4M guanidine contained antibodies that bound to the cartilage surface. By Western blotting, rat antibodies identified 50–65-kd protein bands present in the guanidine extract, but not present either in the material obtained from brief digestion of cartilage with neutrophil elastase or in synovial fluid. The rabbit antisera identified a broad 70–95-kd band. Exposure of elastase-treated cartilage to the guanidine-extracted material resulted in a partial decrease of anti-CII antibody binding. Conclusion. These results suggest that CII on intact cartilage is protected from antibody binding, and that the protective material is at least partly proteinaceous in nature, is unlikely to be derived from synovial fluid, is noncovalently bound to the underlying intercellular matrix, and is synthesized by resident chondrocytes. Further characterization of the protective material may provide important information on the mechanisms of early cartilage damage in inflammatory arthritis.

Objective. To study the interaction of synovial fluid (SF) immunoglobulins with living chondrocytes, and to evaluate the relative contribution of type II collagen (CII) antibodies. Methods. SF of patients with rheumatoid arthritis (RA), osteoarthritis (OA), and gout were incubated with isolated bovine articular chondrocytes. Ig binding was measured by flow cytometry and by quantitation with 125I-labeled anti-IgG and anti-IgM. Complement-dependent cytotoxicity was determined by 51Cr release. Immunoglobulin binding and cytotoxicity were compared between chondrocytes obtained from the superficial and from the deep cartilage zones. Results. Significantly greater IgG and IgM binding was found with RA SF compared with OA or gout SF. Chondrocytes bound more Ig than did fibroblasts. The relative contribution of anti-CII antibodies to Ig binding was studied following absorption of the SF with bovine CII, and by incubation with bacterial collagenase-treated chondrocytes. There was a small but significant reduction in IgG and IgM binding with SF samples that were positive for anti-CII. RA SF exhibited modest, but significantly greater complement-dependent cytotoxicity than OA SF. Gel chromatography fractionation indicated that IgM antibodies were responsible for the cytotoxic activity. Additional studies showed that SF IgM antibodies bound preferentially to, and killed chondrocytes obtained from, the superficial layers of cartilage. Conclusion. Anti-CII antibodies contained in RA SF represent one of many antibody specificities reacting with chondrocyte membrane antigens. Chondrocyte-reactive SF antibodies may play an important pathogenic role in the processes leading to irreversible cartilage damage in RA. These deleterious effects appear to be exerted particularly on chondrocytes located near the articular surface of cartilage.

OBJECTIVE To investigate the interactions between anticollagen antibodies and living chondrocytes. METHODS Mouse monoclonal anti-type II collagen (anti-CII) antibodies, rabbit anti-human CII, and rat anti-CII, anti-CIV, anti-CV, anti-CVI, and anti-CIX were studied in vitro to determine their ability to bind to the plasma membrane of living bovine chondrocytes. RESULTS Mouse monoclonal anti-CII, rabbit anti-CII, and rat anti-CII, anti-CV, and anti-CIX were shown to bind in vitro to the plasma membrane of bovine chondrocytes. Antibody binding was not observed with anti-CIV, or with chondrocytes previously incubated with bacterial collagenase. A significant increase in chondrocyte caseinase and collagenase secretion was observed following sequential incubation with the monoclonal antibodies and a source of activating cytokines. CONCLUSION These results suggest that collagen autoantibodies may exert some of their pathogenic effects on cartilage through interactions with resident chondrocytes, leading to modulation of the rate of secretion of cartilage matrix-degrading enzymes.

GM1-gangliosidosis is a disease characterized by abnormal accumulation of GM1-ganglioside in the brain and viscera. The disease is characterized by clinical findings similar to Hurler's disease and pathologic features resembling Niemann-Pick's disease but with involvement of the glomerular epithelium. A 14-month-old boy, clinically diagnosed as GM1-gangliosidosis, died of respiratory insufficiency and was autopsied except for the brain. Biochemically, marked increase of GM1-ganglioside in the viscera was demonstrated. Pathologically, the foam cells were present in the viscera. Some parts of the cytoplasmic vacuoles in the lungs and spleen contained osmiophilic fibrillar material electron-microscopically. This case was characterized by marked accumulation of foam cells in the pulmonary alveolar spaces.

Eleven autopsy cases (average age of 71 years) with complete right bundle branch block were examined histologically with serial sectioning of the conduction system. The results were as follows: 1) moderate to severe reduction of conducting cells in the right bundle branch (RBB) by fibrous replacement in all 11 cases, 2) a preponderance of main lesions in the first portion of the RBB in cases with R configuration in leads V1 and V2, and in the second to the third portion without significant lesion of the Purkinje cell network in the right ventricle in cases with rsR', rSR', and RSR' configuration in these leads, 3) as to the etiology, 1 case was assumed to have had rheumatic myocarditis and an another case, 1 chronic ischemia. In cases in which the cause could not be clearly established, reduction of the conducting cells in the first portion of the RBB was thought to be due to fibrosis of the upper ventricular septum. Lesions in the second portion of the RBB may be due to mechanical strain effected by muscle crossing of the trabecula septomarginalis and RBB.

Summary The effects of histamine, 5-hydroxytryptamine (5-HT) and prostaglandin E2 (PGE2) on plasma protein extravasation in the rat subcutaneous air-pouch have been studied. Both histamine and 5-HT produced increases in plasma protein extravasation which were inhibited by specific receptor antagonists. Plasma protein extravasation induced by PGE2 was partially inhibited by either a 5-HT receptor antagonist (methysergide) or by a combination of H1 and H2 receptor antagonists (mepyramine and Cimetidine). A combination of all three antagonists further reduced plasma protein extravasation. These results suggest that PGE2 increases vascular permeability indirectly via the degranulation of mast cells. This supposition was confirmed by histological evidence of extensive mast cell degranulation following the injection of PGE2 but not following histamine, 5-HT or saline injection. Using a technique of vascular labelling, following the intravenous injection of Monastral blue dye, plasma extravasation induced by histamine, 5-HT or PGE2 was observed to be restricted to post-capillary venules and was not observed in arterioles or capillaries. Electron microscopic examination of the tissue revealed the presence of monastral blue particles trapped between endothelial cells. These findings suggest that the microcirculation of the rat subcutaneous air-pouch behaves in an analogous manner to that of other tissues.

Microscopic studies of the pulmonary trunk in pulmonary hypertension of 28 cases and control groups of 65 cases ranging from the age of 1 to 83 years were carried out.The elastic configuration of the media of the pulmonary trunk was classified into 5 types and relationship between the severity of the pulmonary hypertension and changes of the elastic fibers in the media was carefully studied.As a result, increase of the elastic fibers even in the acquired severe pulmonary hypertension was observed.This finding is compatible to the views of Meyer although the methods of observation differ and is contradictory to the conclusions of Heath and Saldaiia.Marked changes of the elastic fibers in the media of pulmonary trunk was not found in dogs injected with lycopodeum spores, which may be due to the short duration of pressure loading.A close relationship of the elastic fibers with the smooth muscle cells was observed in the electron microscopic studies of animal experiments.

Summary Background The incidence of postoperative pancreatic fistula (POPF) after distal pancreatectomy is approximately 30%. The most serious complications of pancreatic resection, such as mortality and prolonged hospitalization, are unresolved despite the proposal of various surgical procedures. We developed a new polyglycolic acid (PGA) fabric composed of fine diameter fibers to prevent POPF, and macroscopically and microscopically evaluated the effects of applying it to the pancreatic remnant. Methods The ventral pancreatic surface was cauterized to create the experimental model of POPF in 33 female Wistar/ST rats. The injured sites were wrapped with nonwoven PGA fabrics of different fiber diameters and porosities in the treated rats; one group of rats remained untreated. Survival, incidence of generalized peritonitis, and microscopic findings around the pancreas were investigated. Results The PGA fabrics acted as a scaffold for tissue repair and resulted in superior survival. Generalized peritonitis was milder in the PGA treated groups. With the new PGA fabric, abundant fibroblast infiltration and a uniformly-developed, self-organized barrier wall prevented both pancreatic leak and spread of inflammation. Conclusion Application of the newly developed PGA fabric to the pancreatic remnant prevented POPF, and the essential factor for preventing pancreatic leak was the early formation of a self-organized barrier.

Summary Background/objective Uncontrolled surgical bleeding is associated with increased morbidity, mortality, and hospital cost. Topical hemostatic agents available today have problems controlling hemostatic effects; furthermore, their handling is difficult and they are unsafe. Methods We devised a new hemostatic agent comprising gelatin sponge and film designed to be applied to the bleeding site, thereby creating a topical hemostatic agent made of gelatin alone. The gelatin was prepared by alkali treatment to eliminate viral activity. Hemostatic effects, surgical handling, and tissue reactions of the materials, namely a two-layer sheet of gelatin, TachoSil, and gelatin sponge, were evaluated using 21 dogs' spleens. Results The two-layer gelatin sheet and gelatin sponge exhibited superior hemostatic effects (100% hemostasis completed) compared with TachoSil (0–17% hemostasis). The gelatin matrix immediately absorbed blood flowing from wounds and activated the autologous components in the absorbed blood that promoted coagulation at the bleeding site. The two-layer gelatin sheet had the best surgical handling among the evaluated materials. Materials made of gelatin were associated with fewer inflammatory reactions compared with materials of TachoSil. Conclusion The two-layer sheet of gelatin is a useful topical agent because of its superior hemostatic effects and usability, and is associated with a lower risk of transmitting diseases and inflammatory reactions.