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Biomedical subjects

Y Yamane

Publications and source records attributed to Y Yamane.

393 records · Page 22Linked to original sources

Thermal cross-linking for biologically degradable materials. Preliminary report.

To diminish undesirable side effects of chemical cross-linking of biodegradable materials, the authors developed a thermal cross-linking method that involved esterification by dehydration under dry conditions. The optimal condition for cross-linking was heating at 130-140 degrees C for 40 hr. Efficacy of the cross-linking was evaluated using enzymatic digestion of 0.01% protease in phosphate buffer at pH 7.4. Collagenous materials without cross-linking were digested completely within 30 min. However, with thermal cross-linking, it required 7 days for digestion. The capacity for holding onto water also was tested. Water content decreased when the cross-link density was increased. As an in vivo experiment, six succinylated collagen sealed vascular grafts treated with thermal cross-linking were implanted in the abdominal aortae of dogs and removed 21 days later. These grafts showed no foreign body reaction, and the collagen layer was almost completely absorbed. A collagen sealed graft cross-linked with formaldehyde used as a control showed a strong foreign body reaction. These results suggest that the physical cross-linking method was suitable for biodegradable biologicals, such as collagenous materials, without the undesirable side effects of chemical cross-linking regents.

Animals↗

Cardiovascular hemodynamics and vasopressin blockade in DOCA-salt hypertensive rats.

In conscious rats with near-malignant phases of DOCA-salt (DS) hypertension, hemodynamics were studied with microspheres before and after administration of a vasopressin (VP) vasopressor antagonist in relation to plasma VP levels (pVP). Compared to the controls, the DS rats showed significant elevations in mean arterial pressure (MAP), total vascular resistance (TVR), and pVP, and a flow redistribution from kidney and spleen to skeletal muscles and heart, with increased vascular resistance in almost all organs. The antagonist elicited no significant systemic hemodynamic effects in DS rats as a whole; however, two subgroups, responders vs nonresponders, were identified according to the effects on MAP. In responders with a pVP of 29.2 +/- 2.7 (SE) pg/ml, the antagonist lowered MAP (-24.9 +/- 5.9 mm Hg) and TVR significantly, while in nonresponders with a pVP of 15.2 +/- 3.4 pg/ml, there were no effects. The major antagonist-induced regional responses were increased flow and decreased vascular resistance in skeletal muscles and skin in whole DS rats, and additionally in the gastrointestinal tract, portal organs, and testes in the responders. Significant correlations were observed between pVP, MAP, TVR, and depressor responses to the antagonist only when all data for DS and control rats were pooled. Thus, the systemic hemodynamic effects of VP are important only in responders with exceedingly elevated pVP. VP contributes significantly to the regional hemodynamic abnormalities in skeletal muscles and skin in whole DS rats, and also in several other organs in the responders.

Animals↗

Accuracy of a portable blood gas analyzer incorporating optodes for canine blood.

The accuracy of a portable blood gas analyzer (OPTI 1) was evaluated using canine blood and aqueous control solutions. Sixty-four arterial blood samples were collected from 11 anesthetized dogs and were analyzed for pH, partial pressure of carbon dioxide (PCO2) partial pressure of oxygen (PO2), and bicarbonate concentration ([HCO3-]) values by the OPTI 1 and a conventional blood gas analyzer (GASTAT 3). The conventional analyzer was considered as a standard against which the OPTI 1 was evaluated. Comparison of OPTI 1 results with those of GASTAT 3 by linear regression analysis revealed a high degree of correlation with the GASTAT 3 (r = .90-.91). The mean +/- SD of the differences between OPTI 1 and GASTAT 3 values was -0.008 +/- 0.017 for pH, -0.88 +/- 3.33 mm Hg for PCO2, 3.71 +/- 6.98 mm Hg for PO2, and -0.34 +/- 1.45 mEq/L for [HCO3-]. No statistically significant difference was found between the OPTI 1 and the GASTAT 3. Agreement between these 2 methods is within clinically acceptable ranges for pH, PCO2, PO2, and [HCO3-]. The coefficients of variation for measured pH, PCO2, and PO2 values of 3 aqueous control solutions (acidic, normal, and alkalotic) analyzed by the OPTI 1 ranged from 0.047 to 0.072% for pH, 0.78 to 1.81% for PCO2, and 0.73 to 2.77% for PO2. The OPTI 1 is concluded to provide canine blood gas analysis with an accuracy that is comparable with that of conventional benchtop blood gas analyzers.

Acid-Base Imbalance↗

Sealing of highly porous fabric vascular prostheses by adipose connective tissue fragments instead of preclotting with fresh blood.

Preclotting is an essential procedure for porous fabric vascular prostheses, but fatal bleeding due to fibrinolysis after implantation can occur in some cases. To overcome this problem, a method was developed to seal highly porous fabric vascular prostheses with adipose connective tissue fragments. A piece of subcutaneous adipose connective tissue weighing approximately 1 g was minced with scissors and stirred into 20 mL of saline, thereby creating a tissue suspension. This was sieved through the wall of a highly porous fabric prosthesis from the outside to the inside by intraluminal suction. The prostheses were implanted in the thoracic descending aortae of 20 dogs and removed from 1 h to 1 year after implantation. Twelve preclotted prostheses were used as controls. None of the the treated grafts experienced bleeding complications postoperatively. In the controls, the chest wall was closed after the bleeding stopped from the suture line and through the prosthesis wall, but problems occurred after surgery. Three out of 12 control animals (25%) bled in the pleural cavity within 24 h. In the tissue-sealed grafts, smooth neointima formation without any degenerative changes was observed during a 1-year observation period.

Adipose Tissue↗

Relationship between loss of heterozygosity at microsatellite loci and computerized nuclear morphometry in hepatocellular carcinoma.

Loss of heterozygosity (LOH) at 4 microsatellite loci (chromosomes 8p23, 8q24, 13q14 and 17p13) and the nuclear morphometric data were analyzed in 28 tumors from 28 patients with hepatocellular carcinoma (HCC). LOH was detected in 63.2% of the tumors with marker D8S264 (chromosome 8p23), in 52.6% with marker D8S1801 (chromosome 8q24), in 25.0% with marker D13S328 (chromosome 13q14), and in 52.4% with marker D17S796 (chromosome 17p13). LOH on 17p13 was most frequent in tumors that had reached more advanced stages (p < 0.05). The frequencies of LOH on 17p13 were significantly higher in tumors with portal invasion of cancer cells or intrahepatic metastasis than those without (p < 0.05). Moreover, capsule infiltration of cancer cells or hepatitis B viral infection were more frequently detected in patients with LOH on 17p13 in their tumors. The mean nuclear area (MNA) and the standard deviation (SDNA) of tumors with LOH on 17p13 were significantly larger than those of the other sites (p < 0.01). No significant relationships were found between the LOH on 8p23, 8q24 or 13q14 of HCC and the clinicopathologic data, MNA or SDNA. Therefore, LOH on 17p13 of HCC correlates with the stage, portal invasion of cancer cells, intrahepatic metastasis, and nuclear morphometry. These findings suggest that the LOH on 17p13 is closely connected to the progression of HCC and that nuclear morphometry is useful as an indicator of genetic alterations in HCC.

Adult↗

Establishment of an enzyme-linked immunosorbent assay with a coated deflagellated Salmonella enteritidis antigen for detection of a specific chicken antibody.

We attempted to establish an enzyme-linked immunosorbent assay (ELISA) for field monitoring/profiling purposes for Salmonella enteritidis (SE) infection of poultry flocks. Serotyping rabbit sera, commercially obtained, specific for Salmonella identification sera to O2, O4, O7, O8, S. Vi, S. Hm, and O9, showed negative ELISA (E)-values (< 0.2) on ELISA, except the O9 identification serum (E-value > 0.5). Similar negative E-value results were obtained for antisera to Echerichia coli (E. O antigen). Field serum samples originating from SE-isolated flocks yielded similar positive ratios on both ELISAs including the present coated deflagellated SE antigen and a commercially obtained flagellated SE antigen and that of rapid plate aggregation with a pullorum antigen (PD-RPA). About 100 days after the first monitoring, no SE isolation in the same flock was observed resulting in a carrier state of SE infection. Although both the monitoring results with commercially obtained ELISA and PD-RPA showed lower positive or negative ratios, the present ELISA showed a higher positive ratio than that of the first monitoring. The present ELISA is suggested to be a suitable method to do accurate profiling on the carrier state of infection.

Animals↗

A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin.

In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples.

Animal Husbandry↗

Development of a new scrim cardiac wall substitute.

A cardiac wall substitute made of non-woven ultrafine fibers was developed. It consists of a basal knitted scrim with strongly entangled ultrafine polyester fibers, lined with a fine velour of entangled ultrafine fibers that provide high ravel and tear resistance, a perfect matrix for preclotting, and an anchor for cell adhesion. Six of the new patches and all control patches (a polytetrafluorethylene patch, woven Dacron patch, and glutaraldehyde-treated equine pericardium) were implanted in the right ventricular outflow tracts in 24 dogs. Animal experiments showed that the new patch was easy to handle and suture without fraying or tearing through the edge.

Animals↗

Healing process of vascular prostheses seeded with venous tissue fragments.

Rapid neointima formation in fabric vascular prostheses seeded with autologous venous tissue fragments was examined. A piece of peripheral vein was minced into small fragments and stirred into 20 ml of saline. This tissue suspension was sieved through the wall of Dacron prostheses. The prostheses implanted in the descending aortae of dogs showed extremely rapid healing of the neointima. Endothelial cells lined the entire luminal surface within 14 days. There was no difference in the healing process between the area near anastomotic sites and the center.

Animals↗

Development of a soft, pliable, slow heparin release venous graft.

To prevent their collapse, a certain amount of stiffness is generally required for prosthetic venous grafts, so EPTFE grafts have been used. However, the native vein is pliable without any stiffness. We developed a soft and pliable graft that can maintain patency of the lumen because of its compliance. Fresh porcine ureter was incubated in a ficin solution to remove cell components and noncollagenous proteins. One percent protamine sulfate solution was injected into the ureter lumen to impregnate the inner surface. The ureter was then crosslinked with a 1% glutaraldehyde solution, dipped into a 1% heparin solution for 5 hours, and rinsed with distilled water. This procedure made the ureter very soft and pliable, and also conferred antithrombogenicity to the graft by heparinization. The grafts were implanted into the posterior vena cavae of 20 dogs and were removed from 1 to 878 days after implantation. Eighteen grafts were patent, but two grafts were occluded at the anastomotic site at 218 and 107 days, respectively. As a control experiment, nonheparinized grafts were implanted into 15 dogs; all were occluded with fresh thrombi. All the patent grafts kept their original elasticity, which allowed them to heave in unison with the heartbeat, and were similar in appearance to the native vena cava. Heparinization was effective in preventing thrombus formation. These results indicate that this type of graft is an ideal prosthesis as a venous graft, having physiologic properties such as compliance and antithrombogenicity.

Animals↗

Endothelialization of vascular prostheses by transplantation of venous tissue fragments.

A method to accelerate the endothelialization of vascular prostheses by seeding venous tissue fragments was developed. A piece of peripheral vein was obtained, chopped into small fragments, and stirred into 20 ml of saline, making a tissue suspension. This suspension was sieved through the wall of a highly porous vascular prosthesis (water porosity: 3,600-4,000). The prostheses, (7 mm ID and 5.7 cm in length) seeded with tissue fragments, were implanted into the thoracic descending aortae of 20 dogs, and were removed from 1 to 371 days after implantation. Ten prostheses, preclotted with fresh blood, were used as controls. In the seeded grafts, an infinite number of endothelial cells migrated and proliferated from the fragments. These had produced numerous capillaries by 5 days after implantation that had reached and opened onto the luminal surface of the prosthesis. From these openings, numerous endothelial cells spread out and formed colonies. With the increase in the size of the colonies, the inner surface was completely endothelialized within 5 weeks. This quick neointimal formation by seeding venous tissue fragments might be applicable to several artificial organs.

Animals↗

Development of a growable vascular graft.

A vascular graft that can grow with the growth of its recipient was developed. The graft implanted in the thoracic descending aorta grew slowly to the expected size within a year after implantation in puppies. Human saphenous vein was used as the substrate material. It was dipped into distilled water and sonicated, resulting in cell destruction, and followed by cross-linking with a polyepoxy compound to give both controlled biodegradability, hydrophilicity, and antithrombogenic properties. Four millimeter inner diameter (ID) grafts, enveloped with polyester mesh tubes of 10 mm ID, were implanted in 11 puppies. The diameter of the grafts grew to 9.5 mm from their original 4 mm. After 1 year, the graft walls that were reinforced with polyester mesh were covered with endothelial cells. The following requirements were provided in a growable graft: 1) antithrombogenicity in a small caliber graft; 2) ability to grow as well as to terminate growth. The polyester mesh tube, which was larger than the graft, caused arrest of growth at the expected diameter, whereas the growth rate was controlled by the degree of graft cross-linking. With this method, any size graft can be made by changing the size of the original graft and the polyester mesh tube around it.

Animals↗

A biodegradable antiadhesion collagen membrane with slow release heparin.

A biodegradable antiadhesion collagen membrane with slow heparin release was developed. It was totally antiadhesive on the large intestinal surface in an animal study. Four months after implantation, the membrane was absorbed and the wound surface was covered with mesothelial cells. In general, deposition of fibrin on a wound surface leads to the formation of fibrous adhesions. Therefore, heparin, which inhibits the precipitation of fibrin, is effective in preventing these adhesions. Human amnion was cross-linked with a polyepoxy compound after the impregnation of protamine sulfate into its collagen matrix and was heparinized. Nine adult mongrel dogs were used, five for the heparinized experiment and four as controls. In each animal, two 5 X 5 cm2 areas of the serosa were peeled from the large intestine. The exposed areas were covered with the membrane. The membrane showed an excellent antiadhesion effect in both areas in all five dogs (10 areas). In controls, no membrane was used. The greater omentum, uterus or urinary bladder adhered in five of eight areas. An antiadhesion membrane for soft tissue must not interfere with wound healing as well as prevent adhesions.

Amnion↗

A functional neointima with regularly arranged smooth muscle cells in a fabric vascular prosthesis transplanted with autologous venous tissue fragments.

Regular arrangement of smooth muscle cells underneath an endothelial cell layer was observed in the neointima of a fabric vascular prosthesis treated with new technology to accelerate endothelialization, i.e., transplantation of autologous venous tissue fragments in the graft wall. This finding indicated that the neointima has a vital function as the intima of the blood vessel. A canine left jugular vein was minced and stirred into 20 ml of saline containing 1,000 IU heparin. It was injected with pressure into a fabric prosthesis (4 mm inner diameter [ID], 3.5 cm in length, Water porosity: 4,000 ml) to create the tissue fragmented, heparinized graft. The graft was implanted into the same animal from which the jugular vein was taken. Forty tissue fragmented heparinized (TFH) grafts were implanted in both carotid arteries of 20 dogs and explanted from 1 hr to 400 days after implantation. In this study, the neointimae of the grafts implanted for more than 1 month are analyzed, with a focus on the arrangement of smooth muscle cells in the neointima. A circumferential arrangement of smooth muscle cells with a thin layer of longitudinally arranged cells underneath was seen in the neointimae, which resemble the arrangement of smooth muscle cells in the natural arterial wall. Some areas had a thin smooth muscle cell layer in the longitudinal direction just under the endothelial cell layer. At anastomotic sites, they ran in parallel rows in the longitudinal direction. The authors previously clarified that the smooth muscle cells arrange in parallel rows in the direction of strain caused by tensile stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗