Kim, Young-Suk;Kim, Myung-Jin;Lee, Soo-Jin;Lee, Jae-Il;Jun, Moo-Hyung;Park, Chang-Sik;Kim, Myung-Cheol
Journal of Veterinary Clinics
/
v.24
no.3
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pp.300-304
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2007
The purpose of this study was to determine the anesthetic effects of tiletamine-zolazepam (TZ) alone and azaperone plus tiletamine-zolazepam in growing pigs, and to compare the various physiological parameters in both treatments. Cross experiment was accomplished at 2-week interval. Group 1 (TZ group): six pigs ($31.4{\pm}4.83$ kg) received 4.4 mg/kg of TZ alone. Group 2 (ATZ group); the same six pigs ($43.6{\pm}4.31$ kg) received 4.4 mg/kg of TZ twenty minutes after receiving 2 mg/kg of azaperone. All of the anesthetic drugs were injected into the trapezius muscles. The pigs were fasted for 24 hours before the experiments. Induction and recovery values were determined. Heart rate, respiratory rate, rectal temperature, $pO_2,\;pCO_2$ and pH were determined before administration and 5, 25, 45, 65 and 85 minutes after administration. Induction time of ATZ group was more rapid than that of TZ group (p<0.01). During recovery, sternal recumbency time, standing time and walking time of ATZ group were longer than those of TZ group (p<0.01). Heart rate, respiratory rate, $pO_2,\;pCO_2$, and pH did not show especial differences between the two groups. However, rectal temperature was significantly different between the TZ and ATZ group (p<0.05). As a result, ATZ group had a faster induction and a longer duration of anesthesia than TZ group did. Thus, it was concluded that ATZ combination could be usefully used for chemical restraint in pigs.
The aim of the study was to assess the in vitro effect of 808-nm InGaAs diode laser on rabbit articular chondrocyte proliferation and sulphated glycosaminoglycan (sGAG) synthesis in alginate bead. Previous studies revealed either positive or negative stimulatory effects of laser on different types of cells. A 808-nm InGaAs diode laser at 1.0W power output was used to irradiate the rabbit chondrocytes in alginate beads with energy densities of $31J/cm^2$ (G 1) and $62J/cm^2$ (G 2) corresponding to the experimental groups for 10 seconds and 20 seconds, respectively at 24, 48, 72 and 96 hours after seeding. Control group was left untreated. MTT assay was performed at 1 week and 2 weeks after the $1^{st}$ laser irradiation in alginate beads. sGAG synthesis in alginate beads at 1 week and 2 weeks were determined by DMMB assay. Histological evaluation for cellular distribution and sGAG deposition around the cells were performed by alcian blue stain. MTT assay revealed no positive stimulatory effect in cell proliferation in alginate bead. DMMB assay results showed significantly increased sGAG production in G 2 chondrocytes at 2 weeks. Image analysis of alcian blue stained slides also showed significantly higher percentage of positive alcian blue stain in G 2 chondrocytes. This result suggests that 808-nm InGaAs diode laser with 1.0 W power output although cannot stimulate cell proliferation it can increase the cell secretion activity and sGAG deposition in alginate beads.
The cardiopulmonary responses during total intravenous anesthesia (TIVA) between remifentanil/propofol infusion and remifentanil/ketamine infusion in dogs were compared. Fourteen healthy adult beagle dogs were premedicated with acepromazine (0.1 mg/kg, SC) and medetomidine (20 ${\mu}g$/kg, IV), and anesthetized for 3 hr with remifentanil (0.5 ${\mu}g$/kg/min)/propofol (loading dose: 1 mg/kg, CRI: 0.3 mg/kg/min) CRI (group 'P') or remifentanil/ ketamine (loading dose : 5 mg/kg, CRI: 0.1 mg/kg/min) CRI (group 'K'), respectively. Hemodynamics, blood gas analysis and behavioral changes during recovery were measured. The level of anesthesia was determined by toe-web clamping test. The level of surgical anesthesia was maintained throughout the experiment in both groups. Systolic arterial pressure, mean arterial pressure, $PaO_2$ and $SpO_2$ in group 'K' were significantly higher than in group 'P', and were maintained near the normal ranges. In addition, $PaO_2$ in group 'K' was significantly lower than in group 'P'. However, diastolic arterial pressure, heart rate and respiratory rate were not significantly differed. Mean extubation time from the end of infusion was significantly reduced in group 'K', but mean sitting time was significantly reduced in group 'P'. Mean head-up time and mean walking time were not significantly differed. In group 'K', brief muscle rigidity, head waving and licking during recovery were observed. In conclusion, infusion rate of ketamine (0.1 mg/ kg/min) with remifentanil (0.5 ${\mu}g$/kg/min) is an appropriate for obtaining the surgical plane of anesthesia. These results showed that group 'K' had better cardiopulmonary function than group 'P'. That is, remifentanil/ketamine CRI is better TIVA protocol than remifentanil/propofol CRI for 3 hr surgery.
Natural orifice transluminal endoscopic surgery is a newly emerging technique recently, with its many potential advantages in clinical practice. Cholecystectomy by Hybrid NOTES in this work, performed with single working channel endoscope in conjunction with a laparoscopic grasping forceps in dogs, is a "bridge" between laparoscopic procedure and pure NOTES. Three different approaches for cholecystectomy were carried out; transgastric, transcolonic and transvaginal. In all three approaches, abdominal opening was made by a 5 mm trocar, followed by making pneumoperitoneum of 4 mmHg with $CO_2$ insufflator. Transgastric cholecystectomy, single working channel endoscope was advanced to the peritoneal cavity through gastric incision in antral region made by endoscopic needle knife. Endoscope was retroflexed to visualize the gall bladder. Transcolonic access, incision for endoscopic entry was done at right ventral wall of descending colon, 15 cm inside from the anus. Incision in transvaginal access was made at right-ventral region, just caudal to the caudal tubercle. With the simple traction by the laparoscopic grasping forceps, good visualization of surgical field was obtained in all three groups. Cystic duct and artery were ligated with endoclips; for complete gall bladder dissection from liver, L-knife was used. Closure of incision sites were done in transgastric and transcolonic cholecystectomy by endoclips, not in transvaginal approach.
Park, Se-Il;Moon, Young-Mi;Jeong, Jae-Ho;Jang, Kwang-Ho;Ahn, Myun-Hwan
Journal of Veterinary Clinics
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v.28
no.5
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pp.486-496
/
2011
A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.
Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.
The purpose of present study was to investigate the possibility to immunize guineapigs and swine against Metastrongylus apri infection by the administration of irradiated infective larvae. Four main experiments were undertaken. Firstly, three groups of infective larvae irradiated at $3{\times}10^4r$, $4{\times}10^4r$, and $5{\times}10^4r$ respectively were inoculated to guineapigs and their immunogenic effects were examined from the clinical, anatomical, and serological viewpoints to decide the optimal dose of X-ray for the atenuation of them. Secondly, the migratory behavior of the larvae irradiated at the optimal dose was compared with that of normal infective larvae. Thirdly, pigs were inoculated with each 5,000 infective larvae irradiated at the optimal dose and the clinical, anatomical and serological responses of them before and after challenge were examined. Fourthly, the heated extract of adult M. apri which had been used as an antigen in the serological examinations was analyzed and compared with that of adult Ascaris suis and of adult Trichuris suis by immunoelectrophoretic method. The results obtained are summerized as follows: 1) The optimal dose of X-ray for the atenuation of the infective larvae which can minimize the pathogenecity but keep the antigenecity of the infective larvae was $5{\times}10^4r$. 2) Guineapigs could become completely resistant to subsequent challenge infection by the administration of 1,000 infective larvae irradiated at $5{\times}10^4r$, without showing any symptom of disease before and after the challenge. 3) There were some indication that guineapigs could acquire complete immunity after they overcome the infection with normal infective larvae. 4) It was shown that, in guineapigs, the $5{\times}10^4r$-irradiated larvae can migrate to the large intestine and mesenteric lymph node within a day, where they stay for as long as 16 days to stimulate the host's immunity. 5) It also was shown that, in guineapigs, the normal infective larvae challenged to resistant guineapigs can migrate to the large intestine and mesenteric lymph node, where they are affected by the immune mechanism of host within 10 days without further migration. 6) Pigs could become partially resistant to subsequant challenge by the administration of 5,000 infective larve irradiated at $5{\times}10^4r$; no clinical symptom occurred after the administration, but milder symptoms of parasitic bronchitis were observable after the challenge infection and fewer number of worms were detected from the lungs at autopsy compared with severe symptoms and much number of worms in control pigs. 7) It was shown that, in pigs, a few of the $5{\times}10^4r$-irradiated larvae can migrate to the lungs, where they stay for as long as 104 days in stunted and sterile states; their body-lengths were short and their uteri developted no eggs. 8) There was evidence that the male larvae were more susceptible to X-ray than the female larvae. 9) Antibodies relating to the administration with $5{\times}10^4r$-irradiated or normal larvae were detected from the sera of both guineapigs and pigs by means of indirect haemagglutination and agar diffusion precipitin tests. Relatively higher antibody titers were recorded by the former test, but precipitin bands were demonstrable only when the positive sera were concentrated in one tenth of original volume in the later one. 10) The antibody titers of pig sera began to rose on 14 days, kept their peak during the period from 14 th day to 21st day, and fell to a low level on 28 days after the administration of $5{\times}10^4r$ or normal infective larvae. 11) A slight increase in gamma globublin of the pig sera occurred following the administration. The gamma globulin level showed a tendency to fluctuate in acordance with the antibody level. 12) A marked eosionophilia occurred in pigs on 7 or 14 days following the administration. The eosinophil count showed the same tendency to fluctuate as the gamma globulin did. 13) It was shown that the serum antibodies detected by the heated extract of adult Metastrongylus apri react crossly with the heated extract of adult Ascaris suis but not with that of adult Trichuris suis in indirect haemagglutination and agar diffusion preciption reactions. 14) The heated extract of adult Metastrongylus apri could he divided into 9 antigenic components by immunoelectrophoresis, one (arc 4) of which was shown to be common to both extracts of adult Ascaris suis and adult Trichuris suis, and the other one (arc 9) to only the extract of adult Ascaris suis.
25 tank-mixed combinations between Haloxyfop-methyl at 0, 0.05, 0.1, 0.15 and 0.2 kg ae/ha and Acifluorfen at 0, 0.082, 0.163, 0.245 and 0.326 kg ai/ha were applicated at post-emergence on the late-season cropped soybean(Glycine max) fields predominated by grasses and assessed on the weeding efficacy. A most abundant weed species on the experimented fields were found at September (25 species). The broadleaved weed species and sedges were successively and proportionately emerged by the control rate of grass species. From the mono-treatment of each herbicide, Haloxyfop-methyl was recognized as a highly selective one between grasses and soybean, and Acifluorfen between broadleaf weeds and soybean, respectively. By combined application with both herbicides a synergistic weeding efficacy was detected on total weed species, however antagonistic or additive on each weed groups. Among other traits of soybean, stem dry weight was influenced by weed competitions during October, number of pods per plant during August and seed yields during whole periods. For most effective and reasonable weeding, no less than 0.4 kg ai/ha of each herbicides should be applicated in combinations.
A constructed sea stream in Yeongdo, Busan, Republic of Korea is mostly static due to the lifted stream bed and tidal characters, and receives domestic wastewater nearby, causing a consistent odor production and water quality degradation. Bioaugmentation of a microbial consortium was proposed as an effective and economical restoration technology to restore the polluted stream. The microbial consortium activated on site was augmented on a periodic basis (7~10 days) into the most polluted site (Site 2) which was chosen considering the pollution level and tidal movement. Physicochemical parameters of water qualities were monitored including pH, temperature, DO, ORP, SS, COD, T-N, and T-P. COD and microbial community analyses of the sediments were also performed. A significant reduction in SS, COD, T-N, and COD (sediment) at Site 2 occurred showing their removal rates 51%, 58% and 27% and 35%, respectively, in 13 months while T-P increased by 47%. In most of the test sites, population densities of sulfate reducing bacterial (SRB) groups (Desulfobacteraceae_uc_s, Desulfobacterales_uc_s, Desulfuromonadaceae_uc_s, Desulfuromonas_g1_uc, and Desulfobacter postgatei) and Anaerolinaeles was observed to generally decrease after the bioaugmentation while those of Gamma-proteobacteria (NOR5-6B_s and NOR5-6A_s), Bacteroidales_uc_s, and Flavobacteriales_uc_s appeared to generally increase. Aerobic microbial communities (Flavobacteriaceae_uc_s) were dominant in St. 4 that showed the highest level of DO and least level of COD. These microbial communities could be used as an indicator organism to monitor the restoration process. The alpha diversity indices (OTUs, Chao1, and Shannon) of microbial communities generally decreased after the augmentation. Fast uniFrac analysis of all the samples of different sites and dates showed that there was a similarity in the microbial community structures regardless of samples as the augmentation advanced in comparison with before- and early bioaugmentation event, indicating occurrence of changing of the indigenous microbial community structures. It was concluded that the bioaugmentation could improve the polluted water quality and simultaneously change the microbial community structures via their niche changes. This in situ remediation technology will contribute to an eco-friendly and economically cleaning up of polluted streams of brine water and freshwater.
Rhizoctonia solani [Thanatephorus cucumeris (frank) Donk], one of the major soil-borne plant pathogens with world-wide distribution, can cause great damages on various crops. In Korea, sheath blight on rice caused by this pathogen is the major concern, and active studies on this pathogen have been performed. However, most of these studies were concerned with pathogenicity of the isolates instead of molecular analyses of different AGs of R. solani. Therefore, in this study, thirty isolates of Rhizoctonia solani collected from various sources were used for the analyses of genetic relationships among themselves and for the rapid anastomosis grouping with RAPD method. As a result, thirty isolates of known and unknown AGs were grouped into five subgroups and each group included AG-1, AG-2, AG-3, AG-4, and AG-5. RS-1 isolate was found to be closely related to AG-5. Isolates RS-4, RS-14, RS-17, and RS-16 were found to be closely related to AG-2-2(III B). Isolate RS-13 was closely related to AG-4, isolates RS-8 and RS-10 were closely related to AG-1(I B), and isolates RS-7 and RS-21 were closely related to AG-2-2(IV). Isolate RS-19 was closely related to AG-1(I C), and isolates RS-3, RS-5, RS-18, RS-6, and RS-15 were found to be closely related to AG-1.
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