Escherichia coli heat-labile enterotoxin (LT) and its non-toxic mutant (LTm) are well-known powerful mucosal adjuvants and immunogens. However, the yields of these adjuvants from genetically engineered strains remain at extremely low levels, thereby hindering their extensive application in fundamental and clinical research. Therefore, efficient production of these adjuvant proteins from genetically engineered microbes is a huge challenge in the field of molecular biology. In order to explore the expression bottlenecks of LTm in E. coli, we constructed a series of recombinant plasmids based on various considerations and gene expression strategies. After comparing the protein expression among strains containing different recombinant plasmids, the signal sequence was found to be critical for the expression of LTm and its subunits. When the signal sequence was present, the strong hydrophobicity and instability of this amino acid sequence greatly restricted the generation of subunits. However, when the signal sequence was removed, abundantly expressed subunits formed inactive inclusion bodies that could not be assembled into the hexameric native form, although the inclusion body subunits could be refolded and the biological activity recovered in vitro. Therefore, the dilemma choice of signal sequence formed bottlenecks in the expression of LTm. These results reveal the expression bottlenecks of LTm, provide guidance for the preparation of LTm and its subunits, and certainly help to promote efficient preparation of this mucosal adjuvant protein.
The objective of this study was to detect the fragments generated during IV (intravenous) catheter injection of contrast medium and drug administration in a clinical setting and removal was performed by experimentally producing a phantom, and to compare the radiography, ultrasonography, and multi-detector computed tomography (MDCT) imaging and radiation dose. A 1 cm fragment of an 18 gage Teflon$^{(R)}$ IV catheter with saline was inserted into the IV control line. Radiography, CT, and ultrasonography were performed and radiography and CT dose were calculated. CT and ultrasonography showed an IV catheter fragment clinically and radiography showed no visible difference in the ability to provide a useful image of an IV catheter fragment modality (p >.05). Radiography of effective dose ($0.2139mSv{\cdot}Gy^{-1}{\cdot}cm^{-2}$) form DAP DAP ($0.93{\mu}Gy{\cdot}m^2 $), and dose length product (DLP) ($201mGy{\cdot}cm$) to effective dose was calculated as 0.483 mSv. IV catheter fragment were detected of radiography, ultrasonography and CT. These results can be obtained by menas of an excellent IV catheter fragment of detection capability CT. However, CT is followed by radiation exposure. IV catheter fragment confirming the position and information recommend an ultrasonography.
Medial patellar luxation in dogs is one of the most common patellar problems presented to the veterinary practitioner. It is observed in toy and miniature breed and the majority of cases is a congenital form. Because of extensor mechanism's instability, it causes deformity and disorder in the growth of the affected limb when the luxation is left without treatment As lameness is not easily detectable in puppies, early diagnosis and correction are essential for therapy. Up to now, there has not been any reports refering to the diagnostic methods and the optimal age for correction in young dogs. Thirteen 45-90 days old puppies, have grade I and/or II medial patellar luxation. Only by palpation, all 13 dogs were diagnosed of patelar luxation. Skyline radiographic view was useful to interpret patellar morphology and depth of trochlear groove only above 60 days old. However, it was difficult to make definite diagnosis patellar luxation. The caudocranial and lateral radiographic view as well as ultrasonographic skyline view were not showed of patellar luxation. 2 puppies had unilateral patellar luxation and 11 puppies had bilateral patellar luxation which more serious on the left than on the right. Only 3 puppies among 11 puppies with bilateral patellar luxation were observed of lameness degree 1. Regardless of grade of patellar luxation and lameness, we performed trochlear chondroplasty using a U-shape sculpture blade to minimize cartilage injury, transposition of tibia tuberosity with No. 1 Supramid to align extensor mechanism and lateral imbrication. After surgery, we examined the operated animal daily for 10 days and on 15, 30 and 60 days after surgery respectively. After operation, pain and fever became normal on 7 days, swelling on 10 days, respectively. On 10 days after surgery, dogs showed normal standing position, and normal walking was observed in 15 days after surgery. In force plate analysis, the operated legs were normal weight bearing at 30 days after operation. After surgery, not only patellar luxation and clinical signs have been gradually reduced but also bone growth have become normal without showing growth physeal plate injury. The survival rate of puppies over 62 days old was 100%, while 42-45 days old 37.5%. The above results suggest that optimal age for surgical correction of congenital medial patellar luxation is recommended over 60 days old. In conclusion, combination of trochlear chondroplasty, transposition of tibia tuberosity, and lateral retinacular imbrication is appropriate for over 60 days old puppies to efficiently correct patellar luxation.
Studies on pathogenicities and developmental stages of Nosema apis (Zander, 1909) were carried out through artificial infection to Nosema free honey bees with various levels of spores isolated from local honey bee colony. The results obtained were summarized as follows: 1. The clinical symptoms were observed as dysentery, enteritis of mid-gut (enlargement and decoloration), crawling posture and shortening of the longevity of worker bees in the rearing honey bee colony inoculated with the spores. 2. Number of spores harvested from laboratory rearing honey bees were progresively increased to 4 weeks after inoculation. The regression equations and coefficients of correlations to various spore levels were as follows in each treatment colony. Colony 1. ($$1,000{\times}10^4spores/ml$$) $$y_{c1}=471{\times}10^{4}x+454{\times}10^4(r=0.65^*$$) Colony 2. ($$500{\times}10^4spores/ml$$) $$y_{c2}=340{\times}10^{4}x+207.8{\times}10^4(r=0.99^{**}$$) Colony 3. ($$100{\times}10^4spores/ml$$) $$y_{c3}=150{\times}10^{4}x+84.2{\times}10^4(r=0.99^{**}$$) Colony 4. ($$10{\times}10^4spores/ml$$) $$y_{c4}=13.8{\times}10^{4}x+13{\times}10^4(r=0.98^{**}$$) 3. Average longevity of worker bees artificially infected with Nosema apis was shortened as 21.7~43.8% compare to the control. (p<.05, p<.01) 4. The spores which were isolated from honey bee colony infected with Nosema disease were ovoid or spherical form, and measured, as a rule, from $4.7{\mu}m$ to $6.1{\mu}m$ (mean $5.3{\mu}m$) in length and from $2.4{\mu}m$ to $3.2{\mu}m$ (mean $2.9{\mu}m$) in width. 5. In the mid-gut of honey bees, the spore was progresively germinated and became trophozoite stage. The trophozoites were grown to meronts and their binary fission were begun. The divided two sporoblasts were developed to the spores which had elastic membrane. The new spores were shed in excreta of honey bees 10~15 day after inoculation at $25{\pm}2$ centigrade. 6. The ultrastructure of spore membrane consisted of three layers, such as, outer, middle and inner layer. The sporoplasm consisting lamellar structure occupied only anterior part of the spore and was often extended to posterior direction where definite vacuoles and a polar filament was able to detect.
A 36 year old blindman, engineer was admitted with chief complaints of hemoptysis, recurrent sore throat, pyoderma in genital organ, uveitis and thrombophlebitis for 10 years. Above the chief complaints were remission or exacerbation during hospitalization. Physicalexamination showed that left radial, ulnar & brachial pulse was not palpable. No bruit or murmur was obtained over the mass. Neurologic examination revealed no significant finding.On admission, chest P-A showed hen egg sized round & oval compact hazy density on left upper lung field. Bronchogram revealed no pathological finding and Lt. tomogram showed well define large,ovoid mass density in the superior mediastinum. Fluoroscopy finding showed nonpulsatile on left upper lung field. Pre-op. aortography was not taken, under the impression of lung Ca. rule out .sortie aneurysm, exploratory operation was performed through the 2nd intercostal space, Lt. It was performed that the mass was ascending sortie aneurysm of saccular type. Direct aneurysmectomy with multiple figure of eight suture were done without any prosthetic graft. Post-op. control I.V.C graphy showed completely obstruction sign. Postopcontrol aortography revealed good surgical result. Final, histopathological answered non-specific sortie aneurysm, saccular type. Post-op. courses were uneventful except mild neurologic disturbance with subclavian steal syndrome and associated with both lower leg pitting edema due to inferior vena cava obstruction. After op, 3 month later, discharged to home, with big systemic problem. Behcet`s syndrome reviewed with related literatures. The coexistence of mouth and genital ulceration with hypopyon mentioned by hippocrates and described by various workers in the early part of this century was first defined as a syndrome by Behcet in 1937. In 1937 Behcet described a chronic relapsing triple symptom complex of oral ulceration, genital ulceration, and ocular inflammation. The place of the syndrome as part of a systemic disorder in now clearer, and the under lying pathology appears to be a vasculitis. The disease runs a- chronic course, blindness being the greatest disability and control nervous system involvement a cause of death. Thrombophlebitis is fairly frequent, france et al [1951] giving an incidence of 25% and Dowling [1961] 12%, superficial thrombophlebitis migrans and thrombosis of large veins, including venae cavae [Thomas, 1947: Boolukos 1960] are recorded. Little attention has been paid to arterial involvement. Mishima et al. [1961] described resection cf an aortic aneurysm in a 38 year old man with Behcet`s syndorme. Mounsey in a clinicopathological conference described a case [Brit, med. J., 1966] of ruptured aortic aneurysm in Bechcet`s syndrome treated by aorto-iliac graft. Also, Shikano and Oshima et al [1963] recorded two aneyrysm of smaller arteries. Unfrequently, aortic aneurysm was presumed to be secondary to osteomyelitis of the lumber spine, though the possible association between aortic aneurysm and Behcet`s syndrome was raised. A further case is reported here, in which ascending aortic aneurysm with Behcet`s Ds. appeared to form part of this generalized disease. This is a case report of surgical experience of Behcet`s Ds. with ascending aortic aneurysm which had nearly all the typical clinical features. Above mentioned and was reviewed with related literatures.
International Journal of Industrial Entomology and Biomaterials
/
v.36
no.1
/
pp.15-24
/
2018
N-glycosylation is an important posttranslational modification that results in a variety of biological activities, structural stability, and protein-protein interactions. There are still many mysteries in the structure and function of N-glycans, and detailed elucidation is necessary. Baculovirus expression system (BES) is widely used to produce recombinant glycoproteins, but it is not suitable for clinical use due to differences in N-glycan structure between insects and mammals. It is necessary to develop adequate model glycoproteins for analysis to efficiently alter the insect-type N-glycosylation pathway to human type. The previous research shows the recombinant alpha 1-acid glycoprotein (${\alpha}1AGP$) secreted from silkworm cultured cells or larvae is highly glycosylated and expected to be an excellent research candidate for the glycoprotein analysis expressed by BES. Therefore, we improved the ${\alpha}1AGP$ to be a better model for studying glycosylation. The modified ${\alpha}1AGP$ (${\alpha}1AGP{\Delta}$) recombinant protein was successfully expressed and purified by using BES, however, the expression level in silkworm cultured cells and larvae were lower than that of the ${\alpha}1AGP$. Subsequently, we confirmed the detailed profile of N-glycan on the ${\alpha}1AGP{\Delta}$ by LS/MS analysis the N-glycan structure at each glycosylation site. These results indicated that the recombinant ${\alpha}1AGP{\Delta}$ could be usable as a better model glycoprotein of N-glycosylation research in BES.
HEO Moon-Soo;SONG Choon Bok;LEE Jehee;YEO In-Kyu;JEON You-Jin;LEE Jung-Jae;CHUNG Sang-Chul;LEE Ki-Wan;RHO Sum;CHOI Kwang-Sik;LEE Young Don
Korean Journal of Fisheries and Aquatic Sciences
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v.34
no.4
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pp.365-369
/
2001
Streptococcus spp. of bacterial pathogen of fish were isolated from the cultured flounder (Paralichthys olivaceus) in fish farm of Jeju Island. Clinical signs of the infected flounder which are the most commons symptoms are as follows: erratic swimming, darkening of the body colour, unilateral or bilateral exophthalmia, corneal opacity, hemorrhages in the opercular and the bases of the fins, and the ulceration of the body surface. Biochemical characteristics of pathogenic fish Streptococcus spp, were gram positive, spherical form, catalase negative, oxidase negative and $\beta$-haemolytic, respectively, The viable cells counted from the tissue of the diseased flounder were the largest in the order of the ulcer, the kidney, the blood and the brain, The drugs used were ampicillin, ciprofloxacin, doxycycline, gentamycin, tetracycline, erythromycin, streptomycin and oxytetracycline, Streptococcus spp. were found to be sensitive to ampicillin, ciprofloxacin, doxycycline and gentamycin, but were resistant to tetracycline, erythromycin, streptomycin and oxytetracycline. The pathogenicity of Streptococcus spp. on the cultured flounder with an abdominal cavity injection was high. The haemolytic activity of the toxin against the sheep red blood cells reached the maximum after 30 min incubation at $37^{\circ}C$ or $50^{\circ}C$. The toxin showed highest activity at pH $5.5\sim6.5$.
Temporomandibular joint ankylosis is the movemental obstacle of mandible which depend on proliferation of bony or fibrous tissue in temporomandibular joint structure by various causes. In order to treat this, various surgical methods have been performed, but no operative methods have been produced consistently successful results. This research has been performed to the patients who had been operated due to temporomandibular joint ankylosis by studying classification, cause, onset, duration, anesthesia and treatment method, symptom, change of mouth opening, complication through medical record, X-ray, follow-up for being a help to proper selection of treatment method and evaluation of prognosis. The author obtained the following results by analyzing 44 cases among patients who had been operated due to temporomandibular joint ankylosis during 8 year hospitalization from 1986 to 1993 in Dept. of Oral & Maxillofacial Surgery of Seoul National University Hospital. 1. The occurrence was in the order of below 10, 20's, 10's, 30's. The average of occurrence was 12.95. Illness period was 50.0% within 10 years and 50% beyond 10 years. The average period of illness was 13.33 years. 2. Trauma occupied 54.5% of causes and inflammation occupied 45.5%. Men had more occurrences due to trauma and there was no difference in case of inflammation. 3. In nasotracheal intubations for general anesthesia, the cases of using fiberoptic laryngoscope occupied 40.9%, direct or blind nasotracheal intubation occupied 40.9% and the cases of using tracheostomy occupied 18.2%. 4. In operative approaching methods, submandibular & preauricular approach were mainly applied, and in operative methods, high condylectomy(Group I) occupied 11.4%, arthroplasty without interpositional material following condylectomy or gap ostectomy(Group II) occupied 11.4%, with interpositional material following high condylectomy (Group III) occupied 40.9%, and using condylar reconstruction following condylectomy or gap ostectomy(Group IV) occupied 36.6%. 5. In change of mouth opening reformed after surgery, Group III showed the best result of average 23.5mm, Group IV showed 16.3mm, Group I showed 14.9mm and Group II showed 10.2mm of reformation. Summarizing the results as written above, it is considered that early treatment is important as soon as possible in Temporomandibular joint ankylosis. It is recommended in surgical method what can lead to postoperative early movement maintaining anatomaical & functional form, and then the development of various surgical methods will be requested.
Recent reports indicate that shorter etching times than 60 seconds can be adopted without affecting the bond strength and clinical disadvantages. The purpose of this in vitro study was to compare the shear bone strength and to measure depth of etch at different etching time length. One hundred and eight extracted bovine lower central incisors were embedded each in a tooth cup with cold-cure acrylic resin. The facial surfaces of the teeth were ground wet with 600-, 800-, 1000-, and 1200-grit Sic papers, and finally polished with a water slurry of extrafine silicon carbide powder, washed with tap water, and dried with hot air. Nine groups of nine prepared teeth were etched with a commercial($38\%$ phosphoric acid solution) for 0, 5, 10, 15, 20, 30, 60, 90, and 120 seconds, respectively, rinsed with tap water, and dried with hot air. One conditioned teeth from every group was selected randomly for the scanning electron microscopic examination, and the remaining eight teeth of the groups were used for measuring the push shear bond strength after bonding brackets and immensing them in the $36.5^{\circ}C$ water for 24 hours. Another nine groups of three teeth were used for measuring the depth of etch and surface roughness with a surface profilometer. after pieces of adhesive tape of 3mm inner diameter positioned on the ground enamel surfaces, and etched with the above mentioned. The data obtained form the above expeiments were analysed statistically with one way ANOVA and Dunkan's multiple range test with the $95\%$ confidence level. The results and conclusion of the study were as follows; 1. The results of shear bond strength for the given experimental etching times were not statistically different, but showed the tendency of decreasing shear bone strength after over 60 seconds etching times. 2. On the scanning election microscopic examination, it was observed that the morphological patterns of etched enamel surface for 5 to 20 seconds were similar and consitent, and those for 30 to 120 seconds showed increasing over-etched patterns depending on the length of etching times. 3. The depth of etch was increased almost proportionally by the length of etching times, but it was not associated with the shear bond strength. 4. The surface roughness increased depending on the length of etching times, but it was not associated with the shear bond strength. 5. This experiment indicated that proper etching time with $38\%$ phosphoric acid solution is in the range of 5 to 30 seconds.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.26
no.6
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pp.581-590
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2000
Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Over 30 oncogenes can be activated by insertional mutagenesis, single point mutations, chromosomal translocations and gene amplification. The ras oncogenes have been detected in $15{\sim}20%$ of human tumors that include some of the most common forms of human neoplasia and are known to acquire their transforming properties by single point mutations in two domains of their coding sequences, most commonly in codons 12 and 61. The ras gene family consists of three functional genes, N-ras, K-ras and H-ras which encode highly similar proteins of 188 or 189 amino acid residues generically known as P21. ras proteins have been shown to bind GTP and GTP, and possess intrinsic GTPase activity. Experimental study was performed to observe the mutational change of the ras gene family and apply the results to the clinical activity. 36 Golden Syrian Hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek (control side) was treated with mineral oil as the same manner of the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were completely dissected by microdissection and DNA from both tissue were isolated by proteinase K/phenol/chloroform extraction. Segments of the K-ras and H-ras gene were amplified by PCR using the oligonucleotide primers corresponding to the homologous region (codon 12 and 61) of the hamster gene, and then confirmational change of ras genes was observed by SSCP and autosequencing analysis. The results were as follows : 1. Malignant lesion could be found in the experimental side from the experimental six weeks. 2. One hamster among six showed point mutation of the H-ras codon 12($G{\rightarrow}A$ transition) at the experimental 10 and 14 weeks. 3. One of six at 6 weeks, two of six at 8 weeks and one of six at 12 weeks revealed the confirmational change of the H-ras codon 61($A{\rightarrow}T$ transversion). 4. The incidence of point mutation of H-ras codon 12 and 61 were 5.5%(2 of 36) and 11%(4 of 36) respectively. 5. Point mutation of the K-ras could not be seen during the whole experimental period. Form the above results, these findings strongly support the concept that H-ras oncogenes may have the influence of the DMBA induced carcinoma of hamster buccal pouch.
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