To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGFtreated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.
Getah virus is known as a causative agent of recognized febrile illness of horses characterized by fever, rash and edema. A serological survey indicated that hemagglutination inhibition antibody against Getah virus was detected in 34% of 464 racehorses from Korean Horse Affairs Association and 57% of 262 ponies from Cheju island, respectively. Several field strains of Getah virus isolated were from the racehorse that have been shown fever and febrile signs in 1989. The field isolates produced cytopathic effect in Vero, MA-104, BHK-21 cell cultures. Especially, they multiplied to the highest titer($10^6TCID_{50}/0.1ml$) in Vero cell cultures. When day-old mice were inoculated with field isolates by the intracerebral route, they showed a typical paralysis sign and died within seven days after inoculation. The guinea pig exhibited skin rash and edema, and died with neural signs after inoculation with the field isolates. In the cross neutralization test and indirect immunofuorescent assay, the field isolates were proved to be closely related to the Sakai strain of Getah virus antigenically.
The diagnosis of swine sarcoptic mange (Sarcoptes scabiei var. suis) was investigated by ELISA in order to replace current diagnostic methods such as skin scraping, scratching index, or lesion score of dermatitis. The current methods need many efforts and much times and cost much. They can not handle many samples simultaneously. Therefore, in this research we developed ELISA that can handle many samples at a time. The antigens of swine sarcoptic mite were isolated and examined by 12.5% SDS-PAGE and silver staining. The antigenicity of antigen was confirmed by Western blotting using the swine from the artificailly-infested swine with swine sarcoptic mite. The optical density (OD) values of the artificailly-infested positive sera and the naturally-infested positive sera of sows were measured and read in order to confirm the stability of antigens, the reproducibility and validity (sensitivity and specificity) of the manufactured ELISA of swine sarcoptic antigens. In above results, the developed ELISA would be possible to use the diagnostic tool of sarcoptic mange if OD values of piglets, fattening pigs and sows are interpreted reasonably and classified as mange-free and infested.
The present studies, the investigations were undertaken for an attemp to analyze time course change of biogemic amines in respons to bite injury in rats and guinea pigs, and obtained the following results that might be applicable to the forensic medicine and Forensic Odontology. The histamine concentrations measured at rat abdominal skins were decreased during the first 8 hours and recovered to control levels after 12 hours. However the serotonin concentrations were decreased during the first hours and increased during 4-8 hours, and then returned to control levels thereafter. In contrast with these results, both histamine and serotonin concentrations were tended to increase in guinea pig skins, but the concetration of histamine showed maximum rate of increase immediately after injury and those of sertonin showed maximum rate of increase at 30 minutes. The differencd in the response of histamine and serotonine after injury between rats and ginea pigs revealed that there is specials deffrence in the response of biogenic amines with regard to external stimuli. These results suggest that the determinition of biogenic amines in skin lesion of bite marks could be applied to the estimation of the elapsed time after antemortem injury.
The antigenicity of the aqueous extract of red ginseng (ARG) was evaluated using the following assay procedures : 1. active systemic anaphylaxis (ASA) in guinea pigs, 2 active cutaneous anaphylaxis (ACA) in guinea pigs, 3 passive cutaneous anaphylaxis (PCA) in guinea pigs, 2.active cutaneous anaphylaxis (ACA) in guinea pigs, 3. passive cutanepous anaphylaxis (PCA) in guina pigs with serum for guina pigs sensitized with ARG and 4. passive hemagglutination (PHA) with serum from guinea pigs sensitized with ARG. 1. ASA : No anaphylaxis reaction was observed in any of the sensitized guinea pigs by elictitation with ARG. 2. ACA : No skin reaction was observed in sensitized guinea pigs after intrademal injection of ARG. 3. PCA in guinea pigs : PCA titer of sera from all the sensitized animals was less than 10 in eliciation with ARG. 4. PHA reaction : When eythrocytes coated with challenge antigen were added to sensitized sera, the hemagglutination titer was less than 1. These results suggest that ARG has no antigenicity under the conditions used. And the dose levels of ARG employed in the present experiment were confirmed not to suppress immune reactions.
Kim, Young-Ok;Chung, Hye-Joo;Chung, Seung-Tae;Kim, Jin-ho;Park, Jae-Hyun;Kil, Kwang-Sup;Cho, Dae-Hyun
Archives of Pharmacal Research
/
v.22
no.2
/
pp.143-150
/
1999
melatonin (MLT), N-acetyl-5-methoxytryptamine, is mainly secreted by the pineal gland. The ultraviolet (UV), infrared (IR) and 1H-NMR spectra of irradiated and non-irradiated MLT were measured, and phototoxicity tests of MLT, anthracence (positive control) and sodium lauryl sulfate (SLS, negative control) were performed. The methods employed include both in vitro test such as MTS assay using the human fibroblast cell and yeast growth inhibition assay using Candida albicans and in vivo method using the skin of guinea pig. UV absorption spectra and 1H-NMR spectra of MLT were changed by UVA (365 nm, 15 J/$\textrm{cm}^2$), but IR spectra of MLT were not changed. The fifty percent inhibitory concentration (IC50) ratio (UV-/UV+) of MLT was 10. The inhibition zone of irradiated-paper disks treated with MLT was not observed. According to the results of histophathological examination, no pathologic lesion was observed in the non-irradiated group, but slight degeneration of keratinocytes in the epidermis, homorrhage and vasodilation in dermis were observed in the irradiated group. These results indicated that the molecular structure of MLT is altered by UVA to unidentified photoproducts and a moderate phototoxicity of MLT is predicted.
Various biometerials have been researched and have been developed for treatment of some disease through transplantation to body. They have been evaluated by in vitro cytotoxicity test using some skin-derived cell lines for prediction of their biocompatibility in vivo. However, the results of experiments using mesenchymal or epithelial cells could not be considered in vivo immune reaction. In this study, we evaluated the biomaterial-elution (elute from high density polyethylene film) using some cell lines (L929, Jurkat, U937) in vitro, and then that results were compared with in vivo results from guinea pig sensitization test. In sensitization test, saline and elution of syringe could not induce erythema, but only DNCB (hypersensitive chemical) induce erythema at guinea pig sensitization test. In cell experiment, the cytotoxicity results of inflammatory cells (Jurkat; T lymphocyte, U937; monocyte) was no difference with L929 (fibroblast) in the overall trend. However, inflammatory cell lines were only secreted inflammatory cytokine (TNF-${\alpha}$, INF-${\gamma}$) in some materials (biomateriallution, FAC, DNCB). And the biomaterial-elution did not have toxicity to the cells, but it induced the inflammatory cytokines in inflammatory cell lines only. So, we were predicted inflammatory reaction through the cytokine resultes of inflammatory cell lines, and it was more correlated with in vivo results than cytotoxicity test. Therefore, we suggested that the inflammatory cytokine assay using inflammatory cell lines are more effective method in vitro for evaluation of biocompatibility of biomaterials or chemicals.
Journal of the Korean Society of Fisheries and Ocean Technology
/
v.44
no.3
/
pp.174-183
/
2008
In order to develop the substitutive materials for natural baits of swimming crab pots, the fluorescent characteristics of the baits were analyzed, and the preference of fluorescent dyes were investigated by the mean entrapped catch number to the pots through the water tank experiments and fishing experiments. On the investigation of fluorescent characteristics by the 5 kinds of baits, mackerel, krill, manila clam, pig's fat and chicken's head which were used in substitutive baits for test in the UV long wave(365nm) area, it showed clear blue fluorescence in the skin of mackerel, shell of krill, manila clam and bill of chicken's head, and green fluorescence in the mackerel s muscle and internals, and yellow fluorescence in the pig's fat and chicken's head. On the investigation of fluorescent characteristics by the bait cages in the UV short wave(254nm) and long wave(365nm) area, it showed each green, red and blue fluorescence in the cylinderical or hexahedral red plastic bait cages which were painted each green, red and blue fluorescence dyes, but it showed yellowish green flourescence in the cylinderical or hexahedral red plastic bait cage which was painted yellow fluorescent dye. On the preference investigation of the fluorescent dyes of swimming crabs by the 5 kinds of the bait cages which were put the mackerel in the non-fluorescent red plastic cage($RF_N$), red, yellow, green and blue fluorescent plastic cages(RF, YF, GF, BF) each, nonfluorescent red plastic cage($RF_N$) was entrapped mean 2.0(6.7%), but blue fluorescent plastic cage(BF) was mean 5.0(16.7%) and it was more 2.5 times comparing to $RF_N$, and red fluorescent cage(RF) was same level and green fluorescent cage(GF) was 50% of catch number comparing to $RF_N$, and yellow fluorescent cage(YF) was entrapped nothing(F 46.324, P < 0.05). On the investigation of the entrapped catch number to the pots which were put the mackerel in the blue fluorescent hexahedral plastic cage(HP) and blue fluorescent silicon mackerel model cage(SM), HP was mean 3.4(11.3%) and it was a little more comparing to SM which was entrapped mean 3.2(10.7%)(t 0.775, P > 0.05). Fishing experiments on the preference investigation of swimming crabs by the pots which were put in the non-fluorescent red plastic cage($RF_N$) and blue fluorescent plastic cage(BF) were conducted 3 times. Mean catching number and weight of $RF_N$ were 71.7 ind.(18.3%) and 16.9kg(64.3%), and those of BF were 93.0 ind.(23.1%) and 19.8kg(64.5%), respectively.
We cloned and pharmacologically characterized the guinea pig cysteinyl leukotriene (CysLT) 2 receptor (gpCysLT2). gpCysLT2 consists of 317 amino acids with 75.3%, 75.2%, 73.3% identity to those of humans, mice and rats, respectively. The gpCysLT2 gene is highly expressed in the lung, moderately in eosinophils, skin, spleen, stomach, colon, and modestly in the small intestine. CysLTs accelerated the proliferation of gpCysLT2-expressing HEK293. Leukotriene C4 (LTC4) and Leukotriene D4 (LTD4) enhanced the cell proliferation higher than Bay-u9773, a CysLT2 selective partial agonist and a nonselective antagonist for CysLT receptors. Bay-u9773 did not antagonize the cell proliferation by LTC4 and LTD4. Despite the equipotency of the mitogenic effect among these chemicals, calcium mobilization (CM) levels were variable (LTC4 > LTD4 >> Bay-u9773), and Bay-u9773 antagonized the CM by LTC4. Moreover, the Gi/o inhibitor pertussis toxin perfectly inhibited agonist-induced cell proliferation. These results reveal that cell proliferation via CysLT2 signaling was mediated by Gi/o signaling but independent of calcium mobilization.
This study was conducted to survey the farm which suffered from disease similar to classical swine fever(CSF) in Gyeongbuk province. Clinical signs appeared first in a few number of growing pigs which showed specific signs of diarrhea, depression, sleepiness, and reluctance to get up or to eat. Younger piglets may have appeared chilled, shiver and huddle together. As the disease progresses the affected pig's skin went red and purple. In histopathological signs, there were many haemorrhages throughout the body and larger haemorrhages in some organs such as lymph nodes. And there is a precipitous fall in the number of circulating leukocytes in the blood. In spite of insisting of farmer which did not vaccinate to classical swine fever, significant antibody production was detected in these affected pigs at enzyme-linked immuonsorbent assay. According to the above results at first glance, these affected pig suspected with CSF in clinical signs and histopathological lesions only. Because the symptoms and post-mortem picture were very similar to CSF, these false positive results would have been dangerous to diagnostician. But by reverse transcriptase polymerase chain reaction(RT-PCR) and comparative nucleotide sequence analysis, the disease was correctly diagnosed with post-weaning multisystemic wasting syndrome(PMWS) and porcine reproductive and respiratory syndrome(PRRS) compoundly. And the antigen which were detected the lesion similar to CSF virus, was confirmed with LOM vaccine strain of CSF. In most national CSF eradication program and in countries which are free of the CSF virus, vaccination against CSF is not practiced and generally is not allowed. But now in Korea, routine vaccination is practiced because of outbreaking the CSF repeatedly. When CSF is diagnosed the whole herd and other in contact animal are slaughtered continuously.
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