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PubMed2004

Venous blood flow through vein grafts before harvesting minimizes endothelial cell desquamation immediately after implantation.

AIM: The aim of this paper was to examine effectiveness of maintaining venous blood flow through dissected, but not harvested, grafts on prostaglandin I2 production and endothelial cell regeneration after implantation. METHODS: Using 10 dogs, 8-cm-long bilateral jugular veins were dissected; 1 was harvested, and immersed in heparinized blood for 60 or 120 min. The other was left in situ with retained blood flow until harvest after 60 or 120 min. Two 1-cm-long specimens were obtained from these 4 graft types and subjected to PGI2 assay or silver nitrate staining for measurement of endothelial cell coverage area. The remaining 6-cm portions of the 120-min grafts were autogenously implanted into the abdominal aorta, retrieved after 7 days and assayed and measured. RESULTS: Comparisons between groups suggested beneficial effects of preserving venous blood flow until implantation on endothelial coverage and PGI2 production (p<0.05). CONCLUSION: Venous blood flow through dissected vein grafts effectively helps preserve PGI2 production.

Animals↗
PubMed1992

[New reconstruction method for defect of chest wall using composite graft].

In the case of a 64-year-old man squamous cell carcinoma in the left lung with metastasis to the rib, we excised the affected portion of the chest wall and used a composite graft, a combination hydroxyapatite (HAP) filler, for reconstruction with good results. HAP filler is a ceramic that combines the calcium phosphate apatite hydroxide with 3-calcium phosphate. The Dacron fabric used was made of polyethylene terephthalate fiber. Two layers of Dacron fabric were laid together, stitched to the rib with nylon thread, and the resulting tubular cavity packed with HAP filler to create an artificial rib. The intercostal muscle was sandwiched between the layers of Dacron fabric and the ends of the natural rib laid across those of the artificial one. We used the periosteum and intercostal muscle to integrate the graft and complete reconstruction. Postoperative recovery was good, and there was no visible sign of deformation at the graft site. Even in palpation we were unable to tell that the rib was artificial.

Biocompatible Materials↗
PubMed1983

Identification of the probable site of synthesis of butyrylcholinesterase in the superior cervical and ciliary ganglia of the cat.

The source of butyrylcholinesterase (acylcholine acylhydrolase, EC 3.1.1.8) in the ganglion cells of the cat superior cervical and ciliary ganglia has been elusive, inasmuch as the enzyme is present in high concentrations in the neuropil, where it is confined largely to the dendritic and perikaryonal plasma membranes, but appears to be absent from the perikarya. In the present study, ganglionic butyrylcholinesterase was near-totally inactivated by the injection of tetramonoisopropyl pyyrophosphoramide (6.0 mumol/kg of body weight) intravenously. During the ensuing 72 hr, the regenerating enzyme became detectable by the copper thiocholine histochemical method in the somata of essentially all ganglion cells and in the neuropil. Results were similar in preganglionically denervated superior cervical ganglia and in normal ciliary ganglia. These findings suggest (i) that butyrylcholinesterase indeed is synthesized in the ganglion cell perikarya (presumably, the rough endoplasmic reticulum) and transported extremely rapidly to more peripheral cellular sites and (ii) that the synthesis is largely independent of control by any neurotrophic factor provided by the preganglionic axonal terminals. Similar studies were conducted in the rat. In this species, in contrast to the cat, the somata of essentially all ganglion cells of the superior cervical ganglion contain various but at least moderate concentrations of acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7) and propionylcholinesterase (acylcholine acylhydrolase, EC 3.1.1.8). After injection of tetramonoisopropyl pyrophosphoramide, propionylcholinesterase reappeared in the ganglion cell somata before its accumulation in the neuropil, as would be expected.

Acetylcholinesterase↗
PubMed2005

Matrix metalloproteinase-9 modulation by resident arterial cells is responsible for injury-induced accelerated atherosclerotic plaque development in apolipoprotein E-deficient mice.

OBJECTIVE: Although matrix metalloproteinase-9 (MMP-9) has been implicated in atherosclerotic plaque instability, the exact role it plays in the plaque development and progression remains largely unknown. We generated apolipoprotein E (apoE)-deficient (apoE-/-) MMP-9-deficient (MMP-9-/-) mice to determine the mechanisms and the main cell source of MMP-9 responsible for the plaque composition during accelerated atherosclerotic plaque formation. METHODS AND RESULTS: Three weeks after temporary carotid artery ligation revealed that while on a Western-type diet, apoE-/- MMP-9-/- mice had a significant reduction in intimal plaque length and volume compared with apoE-/- MMP-9+/+ mice. The reduction in plaque volume correlated with a significantly lower number of intraplaque cells of resident cells and bone marrow-derived cells. To determine the cellular origin of MMP-9 in plaque development, bone marrow transplantation after total-body irradiation was performed with apoE-/- MMP-9+/+ and apoE-/- MMP-9-/- mice, which showed that only MMP-9 derived from resident arterial cells is required for plaque development. CONCLUSIONS: MMP-9 is derived from resident arterial cells and is required for early atherosclerotic plaque development and cellular accumulation in apoE-/- mice.

Animals↗
PubMed1987

Correlative studies on antigenicity of pancreatic cancer and blood group types.

Blood group-related antigenicity in 14 pancreatic cancer patients was examined by immunohistological method using monoclonal antibodies (MoAbs) against A, B, H, Lea, Leb, and CA 19-9 and compared with the phenotypic expression of the individuals' blood groups. MoAb-A reacted strongly with tumor tissue in four of five blood group A patients. Two of two patients with blood type B showed a weak, focal reactivity of their cancer with MoAb-B. H antigen was found in four of five patients from blood group A, while it was present in only one blood group O person and absent in B-type individuals. B antigen was inappropriately expressed in one person with type A blood and in two with type O. Lea antigen was expressed in all but two tumor tissues and Leb antigen in all tumorous tissues, irrespective of Le blood group status. MoAb 19-9 reacted with 11 of 14 cases.

ABO Blood-Group System↗
PubMed1994

[Various factors influencing in vitro antifungal activities of omoconazole nitrate (OMZ), a new imidazole antimycotic].

Effects of various factors on in vitro antifungal activities of omoconazole nitrate (OMZ) were investigated with bifonazole (BFZ) as the reference drug using an agar dilution method and a broth dilution method. Composition of the culture medium significantly altered the activity as determined by minimum inhibitory concentration (MIC) of OMZ; OMZ exhibited a greater anti-Candida activity (smaller MIC values) on casitone agar (CA) than on Sabouraud dextrose agar (SDA). Anti-Candida activity of OMZ was the highest in pH 5 medium, but it was lowered by an increase of inoculum size, a prolongation of the incubation period and the addition of calf serum to the medium. Anti-Trichophyton activity of OMZ was not influenced with these factors except for the addition of calf serum. The activity of OMZ was fungicidal against Candida at pH 5.0 and against Trichophyton at pH 5.0 and 6.6. The geometric mean MIC of OMZ was lower than that of BFZ against C. albicans freshly-isolated on acid CA.

Antifungal Agents↗
PubMedDate not supplied

Insulin resistance is associated with reduced nocturnal falls of blood pressure in normotensive, nonobese type 2 diabetic subjects.

To assess the relationship between insulin resistance and ambulatory blood pressure (BP) pattern, we determined glucose infusion rate (GIR) as a marker of insulin resistance using a glucose clamp method, and measured 24-h BPs in 25 normotensive, nonobese type 2 diabetic subjects. They were divided into two groups: 11 dippers and 14 nondippers. Clinical characteristics were similar in the two groups except for orthostatic fall in systolic BP. The median GIR level was significantly lower in nondippers than in dippers (P < 0.05). Spearman's rank correlation revealed that the GIRs were negatively correlated with the systolic, diastolic and mean BPs during nighttime (P < 0.05 or less), but not with daytime or whole day BPs. Moreover, based on a logistic regression analysis, the GIR as well as orthostatic fall in systolic BP discriminated independently between dippers and nondippers. Thus, our results suggest that insulin resistance is associated with decreased nocturnal BP fall in type 2 diabetic subjects.

Adolescent↗
PubMed1996

8-hydroxy-2'-deoxyguanosine is increased in epidermal cells of hairless mice after chronic ultraviolet B exposure.

8-Hydroxy-2'-deoxyguanosine (8-OHdG) is a mutation-prone (G:C to T:A transversion) DNA base-modified product generated by reactive oxygen species or photodynamic action. G:C to T:A transversions are observed in the p53 and ras genes of UVB-induced skin cancers of mice and in squamous and basal cell carcinomas of human skin exposed to sunlight. In the current study, 8-OHdG formation was evaluated in the epidermis of hairless mice after repeated exposure to UVB, and possible mechanisms involved were studied. Exposure of hairless mice to either 3.4 [2 minimal erythema dose (MED)] or 16.8 (10 MED) kJ/m2 of UVB three times a week for 2 wk induced a 2.5- or 6.1-fold increase, respectively, in the levels of 8-OHdG in DNA, compared to the unexposed controls. An immunohistochemical method using a monoclonal antibody specific for 8-OHdG showed stronger and more extensive staining in the nuclei of UV-irradiated epidermal cells than in those of nonirradiated cells. Western blots probed with antibodies against 4-hydroxy-2-nonenal-modified proteins confirmed the involvement of reactive oxygen species in the epidermal damage induced by chronic UVB exposure. 3-Nitro-L-tyrosine was detected in western blots in a concentration-dependent manner, suggesting that peroxynitrite derived from the reaction of nitric oxide and superoxide, both of which were probably released from inflammatory cells, was involved in modifying the DNA bases. Therefore, the formation of 8-OHdG after UVB exposure appears to be regulated by at least three pathways: photodynamic action, lipid peroxidation, and inflammation and may play a role in sunlight-induced skin carcinogenesis.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed2005

Silent cerebral infarction predicts vascular events in hemodialysis patients.

BACKGROUND: Cardiovascular disease is the leading cause of death in hemodialysis (HD) patients. We have previously reported a higher incidence of silent cerebral infarction (SCI) in HD patients compared with the control group using MRI studies. In the present study, we examined whether or not SCI could predict vascular events in HD patients. METHODS: Cranial magnetic resonance imaging (MRI) was performed on 119 HD patients without symptomatic cerebrovascular disease. SCI was detected by MRI, and the patients were prospectively followed up. The end points of the study were the incidence of major events related to vascular events (cerebral events, cardiac events, and sudden deaths). We investigated the prognostic role of SCI in cerebral, cardiac, and vascular events by using the Kaplan-Meier method and Cox proportional hazards analysis. RESULTS: The prevalence of SCI was 49.6% in HD patients. During a follow-up period of maximum 60 months, vascular events, which included 13 cerebral events, 5 cardiac events, and 3 sudden deaths, occurred in 21 patients. The presence of SCI was predictive for a higher cerebral and vascular morbidity compared to the absence of SCI [18.6% (N= 11) vs. 3.3% (N= 2), P= 0.0169, and 30.5% (N= 18) vs. 5.0% (N= 3), P= 0.0006, respectively]. By multivariate Cox proportional hazards analysis, SCI remained a powerful independent predictor of cerebral and vascular events (hazard ratio for cerebral events 7.33, 95% CI 1.27-42.25: for vascular events 4.48, 95% CI 1.09-18.41). CONCLUSION: The findings of the present study indicate that the presence of SCI is a new risk factor for vascular events in HD patients.

Adult↗
PubMed1994

Oral delivery of poly(lactide-co-glycolide) microspheres containing ovalbumin as vaccine formulation: particle size study.

The objective of the present study was to produce ovalbumin (OVA) loaded poly(lactide-co-glycolide) (PLGA) microspheres with different diameters and to evaluate their possibilities as vaccine formulation in mice following oral inoculation. Four kinds of OVA loaded PLGA microspheres with different mean average volume diameters (1.3, 4.0, 7.5, and 14.0 microns) were manufactured using a w/o/w emulsion/solvent evaporation method. Low loading efficiencies (8-20% w/w) were observed in all batches although smooth spherical particles were obtained. Single oral administrations of OVA loaded PLGA microspheres with different diameters to mice produced immune responses (serum IgG levels by ELISA) which were statistically higher than OVA solution as negative control (Fisher's paired t-test). A dose-response was observed, and single and double inoculation orally produced similar serum antibody levels. The rank of immune response was as follows: 4.0 microns- > 1.3 microns- = 7.5 microns- > 14.0 microns-microspheres. The oral inoculation with 0.13% OVA loaded PLGA microspheres having a mean volume diameter of 4.0 microns exhibited the best immune responses with values greater than those obtained after subcutaneous inoculation with complete Freund's adjuvant (CFA) as positive control, and not significantly different from those obtained after subcutaneous inoculation with the same microspheres.

Adjuvants, Immunologic↗
PubMed1993

Functional morphology of the lymphatic system in the monkey diaphragm.

A morphological study of the lymphatic system in the diaphragm of adult Japanese monkeys was carried out using intraperitoneal injection with India ink, light microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) techniques. For SEM examination of the subperitoneal connective tissue, the peritoneal mesothelium was stripped by either the NaOH/sonication microdissection techniques or the NaOH digestion method. Light microscopy of 1 micron Epon sections and TEM of thin sections demonstrated that lymphatic capillaries were distributed in the subperitoneal connective tissue of the muscular and tendinous portions. Gaps (stomata) between neighboring cells of the peritoneal mesothelium were occasionally encountered. In SEM observations, stomata with oval shapes and various sizes were abundantly seen in the peritoneal mesothelium, but were absent from the pleural mesothelium. It deserves particular note that a large number of foramina were detected in the subperitoneal connective tissue. The cluster of the foramina was similar in appearance to the "macula cribriformis" depicted by KIHARA (1956). Carbon particles injected into the monkey peritoneal cavity passed through the peritoneal stomata and then the macula cribriformis, ultimately to enter the lymphatic capillaries. This paper also discusses the functional implication of the lymphatic system in the diaphragm.

Animals↗
PubMed2005

Ultrastructural analysis of the behavior of the dimorphic fungus Microbotryum violaceum in fungus-induced anthers of female Silene latifolia flowers.

The development of male organs is induced in female flowers of the dioecious plant Silene latifolia by infection with the fungus Microbotryum violaceum. Stamens in a healthy female flower grow only to stage 6, whereas those in an infected female flower develop to the mature stage (stage 12), at which the stamens are filled with fungal teliospores instead of pollen grains. To investigate these host-parasite interactions, young floral buds and fungus-induced anthers of infected female flowers were examined by electron microscopy following fixation by a high-pressure freezing method. Using this approach, we found that parasitic hyphae of this fungus contain several extracellular vesicles and have a consistent appearance up to stage 8. At that stage, parasitic hyphae are observed adjacent to dying sporogenous cells in the infected female anther. At stage 9, an increased number of dead and dying sporogenous cells is observed, among which the sporogenous hyphae of the fungus develop and form initial teliospores. Several types of electron-dense material are present in proximity to some fungi at this stage. The initial teliospores contain two types of vacuoles, and the fungus cell wall contains abundant carbohydrate, as revealed by silver protein staining. The sporogenous cell is probably sensitive to infection by the fungus, resulting in disruption. In addition, the fungus accelerates cell death in the anther and utilizes constituents of the dead host cell to form the mature teliospore.

Basidiomycota↗
PubMed1995

Immunohistochemistry of gap junctions in normal and diseased gastric mucosa of humans.

BACKGROUND & AIMS: Intercellular communication through gap junctions has been proposed to be an important mechanism for the maintenance of tissue homeostasis. However, few studies have considered the role of gap junctions in gastric mucosa. The purpose of this study was to evaluate the distribution of gap junction-specific proteins in normal and diseased gastric mucosa of humans. METHODS: Biopsy specimens were obtained endoscopically. Immunohistochemical staining was performed by standard immunoperoxidase techniques using an anti-connexin 32 monoclonal antibody. RESULTS: In normal gastric mucosa, connexin 32 was present chiefly in the foveolar cells and in a decreasing staining gradient extending down to the necks of the glands. Connexin 32 also was observed in the epithelial cells of atrophic mucosa in a pattern similar to that observed in normal controls. Conversely, the majority of the foveolar cells adjacent to erosions had reduced or absent staining. Connexin 32 also was reduced significantly or absent from metaplastic epithelial cells. No malignant cells from patients with carcinoma contained detectable connexin 32. CONCLUSIONS: Intercellular communication likely is impaired in precancerous or paracancerous lesions of the stomach. Abnormal intercellular communication thus may play an important role in the progression from mucosal injury to intestinal metaplasia and/or gastric carcinoma.

Adult↗
PubMed2004

Antitumor effect induced by dendritic cell (DC)-based immunotherapy against peritoneal dissemination of the hamster pancreatic cancer.

Establishing a method to control peritoneal dissemination is one of the most pressing issues in the postsurgical treatment of pancreatic cancer. In the present study, we investigated the effect of dendritic cell (DC)-based immunotherapy on peritoneal disseminations of hamster pancreatic cancer cells, PGHAM-1. After the orthotopically inoculation of 2 x 10(6) PGHAM-1 cells, DC pulsed with PGHAM-1-derived tumor lysates, DC alone or PBS as a vehicle was injected intraperitoneally (i.p.) three times at weekly intervals. The group treated with DC or DC+lysate was found to have smaller disseminated tumors than the vehicle-treated. In addition, mean survival time in the DC+lysate groups was significantly longer than the PBS group. These findings suggested that DC-based immunotherapy might be efficient for the treatment of peritoneal disseminations of the pancreatic cancer.

Animals↗
PubMed2005

Localization and changes of intraneural inflammatory cytokines and inducible-nitric oxide induced by mechanical compression.

STUDY DESIGN: Investigation of intraneural inflammation induced by mechanical compression. OBJECTIVES: In order to investigate the mechanism of neuropathy, this study used a median nerve compression model in dogs. Immunohistochemistry was used to examine the localization and changes of inflammatory cytokines and nitric oxide (NO). SUMMARY OF BACKGROUND DATA: The manifestation of pain at sites of inflammation has a close relationship with the release of mediators from macrophages such as interleulin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha), as well as with NO. However, the mediators involved in inflammation of nerve due to mechanical compression remain almost unknown. METHODS: In this study, the median nerve of dogs was compressed with a clip for three weeks to observe the changes caused by compression. Immunohistochemistry was done by the avidin-biotin-peroxidase complex method to observe the changes of T cells (CD45) and macrophages (Mac-1) after compression. Antibodies against IL-1beta, TNF-alpha, and inducible nitric oxide synthesis (i-NOS) were used to examine the localization and changes of these mediators caused by nerve compression. RESULTS: In control animals, resident T cells were detected, but there were no macrophages. IL-1beta was positive in the Schwann cells and vascular endothelial cells. However, no cells showed TNF-alpha or i-NOS positively. After nerve compression, numerous T cells and macrophages appeared among the demyelinized nerve fibers. The macrophages were positive for IL-1beta, TNF-alpha and i-NOS. CONCLUSION: Inflammatory cytokines and NO may be involved in intraneural inflammatory changes arising from mechanical compression. Such mediators may be of importance in the manifestation of neuropathy.

Animals↗
PubMed2001

Antibody against oxidized low density lipoprotein may predict progression or regression of atherosclerotic coronary artery disease.

OBJECTIVES: This study aimed to elucidate whether an antibody against oxidized low density lipoprotein (anti-Ox-LDL) could predict short-term coronary artery atherosclerotic lesion progression. BACKGROUND: It is still controversial whether higher levels of the anti-Ox-LDL titer are associated with atherosclerotic coronary artery disease. METHODS: In 52 patients undergoing coronary angioplasty and six-month follow-up angiography, we performed quantitative coronary angiographic analysis of a lesion on a branch away from the intervention site vessel and assessed lesion progression or regression using the Progression-Regression score calculated as the baseline minimal lumen diameter minus the follow-up minimal lumen diameter. The serum anti-Ox-LDL titer was measured using an enzyme-linked immunosorbent assay method just before the initial angiography in all patients. RESULTS: The anti-Ox-LDL titer was 16.6+/-1.5 AcU/ml in the progression group (Progression-Regression score >0.15 mm; n = 20), which was significantly higher (p < 0.001) than the value of 9.5+/-1.2 in the regression group (< or =-0.15 mm; n = 14) and also higher (p < 0.01) than the value of 11.4+/-1.3 in the no-change group (-0.15 to 0.15 mm; n = 18). The Progression-Regression score was correlated with the antibody titer in all patients (r = 0.56, p < 0.001). Multiple regression analysis showed that the Progression-Regression score was independently correlated with the antibody titer (r = 0.44, p < 0.01) as well as lipoprotein (a) (r = 0.33, p < 0.05). CONCLUSIONS: Anti-Ox-LDL may be an independent predictor of coronary atherosclerotic lesion progression in the short term.

Antibodies↗
PubMed2001

Dizocilpine but not ketamine reduces the volume of ischaemic damage after acute subdural haematoma in the rat.

BACKGROUND AND OBJECTIVE: Increased glutamate concentration in the cerebrospinal fluid has been reported in severely head-injured patients, suggesting that an excessive release of glutamate may be involved in the process of neuronal damage. Ischaemic damage after subdural haematoma has been reported to be reduced by glutamate (N-methyl-D-aspartate: NMDA) receptor antagonists such as dizocilpine and CGS 19755; even though these drugs were given 20-30 min after insult. Excessive release of excitatory amino acids may produce the neural damage after subdural haematoma and NMDA receptor antagonists may become valuable therapeutic drugs. This study compared the effects of ketamine and dizocilpine, on intracranial pressure and histopathological changes after acute subdural haematoma produced by an injection of autologous blood (150 microL) in rats. METHODS: The control (n = 9), ketamine (n = 9) and dizocilpine (n = 9) groups, respectively, received saline, ketamine (total dose: 210 mg kg-1) or dizocilpine (total dose: 1.0 mg kg-1) from 0.5 to 8 h after acute subdural haematoma. A silicone group (n = 9) had the same volume of silicone injected subdurally. RESULTS: The volume of ischaemic damage in the silicone group (1.3 +/- 1.2 mm3) was significantly smaller than in the control group (11.9 +/- 3.8 mm3). Ketamine and dizocilpine did not increase intracranial pressure. Dizocilpine significantly decreased the volume of ischaemic damage (6.1 +/- 3.8 mm3). Ketamine failed to significantly decrease damage (7.8 +/- 5.0 mm3). CONCLUSIONS: These results suggest that the factors elicited by the clotted blood contribute to the ischaemic damage after subdural haematoma, and that the glutamate receptor antagonist dizocilpine reduces the damage, while ketamine shows only a trend reduction of the damage.

Animals↗
PubMed2001

A simple and sensitive detection system for Bacillus anthracis in meat and tissue.

AIMS: To detect and isolate Bacillus anthracis from meat and tissue by rapid and simple procedures. METHODS AND RESULTS: Bacillus anthracis Pasteur II cells were added to 1 g lymph node and pig meat, which were then cut into small pieces and suspended in PBS. Aliquots were spread on Bacillus cereus selective agar (BCA) plates to isolate B. anthracis cells, and incubated in trypticase soy broth. The enrichment culture was used for nested PCR with B. anthracis specific primers, which were to confirm the presence of B. anthracis chromosomal DNA and the pXO1/pXO2 plasmids. CONCLUSION: One cell of B. anthracis was detected by nested PCR from 1 g of the samples, and was also isolated on BCA plates according to colony morphology within two days. SIGNIFICANCE AND IMPACT OF THE STUDY: These results could be useful for detecting animals with latent anthrax, and meat contaminated with B. anthracis, rapidly and simply.

Animals↗