Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.
The objective of the present study was to investigate the expression of heat shock protein 90 (HSP90) and 70 (HSP70) mRNA as cellular stress responses, the levels of plasma cortisol with glucose as neuro-endocrine stress responses during water temperature rising in freshwater adapted black porgy, Acanthopagrus schlegeli. A cDNA fragment of 891 (HSP90) and 465 (HSP70) bp was cloned from black porgy testis by Reverse transcription-polymerase chain reaction (RT-PCR) with primers designed from the conserved regions of other teleost. The PCR product of HSP90 showed very high homology to red seabream (99%), rainbow trout (95%), Atlantic salmon (94%), zebrafish (94%) HSP90, HSP70 of black porgy was also highly similar to those of rainbow trout (96%), silver seabream (95%), zebrafish (95%) HSP70. Water temperature rising ($20{\sim}30^{\circ}C$) induced elevation of HSP90 mRNA in black porgy gonad, liver, brain, intestine and kidney, whereas it resulted in an induction of the HSP70 mRNA expression in gonad only. Plasma cortisol levels increased significantly at $30^{\circ}C$ in the fish compared to those at $20^{\circ}C$. Glucose levels of the fish showed a tendency of co-increase with cortisol during water temperature rising. These results suggest that increased HSP90 mRNA in liver with plasma cortisol following heat shock may be related to increasing glucose for homeostasis in this species.
Kim, Soung-Min;Lee, Jong-Ho;Jo, Joung-Ae;Lee, Seung-Cheol;Lee, Suk-Keun
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.31
no.5
/
pp.440-453
/
2005
Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.
Purpose : To evaluate the efficacy of arthroscopic management of the triangular fibrocartilage complex(TFCC). Materials and Methods : Thirteen patients(14 wrists) with acute or chronic traumatic triangular fibrocartilage complex lesions were included in the study. The mean patients' age was 28.3 years, with a range of 21 to 45 years. All patients were diagnosed by physical examination, arthrographic or magnetic resonance imaging studies. Eight of the 14 wrists had central TFCC tear while 6 wrists had peripheral tear. Under arthroscopic control, injuries to the central portions were treated by debridement and excision of unstable tissue fragment while peripheral tears were repaired. The follow-up period averaged 28 months. The results were analyzed clinically using the Mayo modification of the Green and O' Brien wrist scoring system. Results : Nine of the 14 wrists were rated excellent,3 good and 2 fair Overall, 12 of the 14 patients rated as satisfactory and returned to sports or work activities. Conclusion : Arthroscopic treatment of TFCC resulted in a high degree of patient satisfaction and an increase in the ability to perform at workshop.
With a double mismatch primer set designed for amplifying the modified DNA sequence fragments, bovine melanocortin-1 receptor(MC1R) gene encoded in Extension locus which plays a critical role in coat color development was analyzed using polymerase chain reaction mediated restriction fragment length polymorphism(PCR-RFLP). Amplified PCR fragments were successfully discriminated with combining the MspI- and AluI-RFLP into three major alleles(ED, E+, and e), directly related to bovine coat color phenotypes. The genotyping results showed that Jeju black cattle contained three MC1R alleles, but yellowish-red colored Hanwoo and bridle colored Korean Brindle cattle did not contained the dominant black allele ED. However, two dominant black-colored cattle breeds, Holstein and Angus, contained the ED allele over 96% in frequency. Hanwoo×Holstein F1 and Hanwoo×Angus F1 crossbred calves showed ED/e MC1R genotypes, and uniformly black coat color. the results suggested that this MC1R genotyping method be useful in allele discrimination for bovine MC1R gene which used for breed classification and characterization, as one of the important genetic markers, using combination of MspI- and AluI-RFLP for modified PCR product amplified with a newly designed double mismatch primer set.
To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.
The optimum blasting pattern to excavate a quarry efficiently and economically can be determined based on the minimum production cost which is generally estimated according to rock fragmentation. Therefore it is a critical problem to predict fragment size distribution of blasted rocks over an entire quarry. By comparing various prediction models, it can be ascertained that the result obtained from Kuz-Ram model relatively coincides with that of field measurements. Kuz-Ram model uses the concept of rock factor to signify conditions of rock mass such as block size, rock jointing, strength and others. For the evaluation of total production cost, it is imperative to estimate 3-D spatial distribution of rock factor for the entire quarry. In this study, a sequential indicator simulation technique is adopted for estimation of spatial distribution of rock factor due to its higher reproducibility of spatial variability and distribution models than Kriging methods. Further, this can reduce the uncertainty of predictor using distribution information of sample data The entire quarry is classified into three types of rock mass and optimum blasting pattern is proposed for each type based on 3-D spatial distribution of rock factor. In addition, plane maps of rock factor distribution for each ground levels is provided to estimate production costs for each process and to make a plan for an optimum blasting pattern.
Purpose: In this study, we tried to develop the technique of osteotomy for hallux valgus. The new modified technique of osteotomy was accomplished with even more greater stability, accurate correction of the deformity and more effective than 'chevron' osteotomy in terms of correction of the deformity. Materials and Methods: Between March 1998 and December 2001, 55 cases of new modified osteotomy for hallux valgus were performed for 39 patients, 16 of whom underwent operation of both feet. Operations were made for 34 women and 5 men whose average age was 46 years old (range, $20{\sim}71$ years). Average follow up period was three years (range, $2{\sim}5$ years), and during the follow up, the patients underwent physical examination and assessment with use of the American Orthpaedic Foot and Ankle Society's hallux-metatarso-phalangealinterphalangeal scale and standard foot radiographic measurements. Results: 37 patients (53 cases) out of 39 patients (55 cases) had no pain, good cosmesis, and all of the patients were satisfied with the results of the operation. Two had occasional mild discomfort. The average score according to the hallux-metatarso-phallangeal-interphalangeal scale was 93.2 points (range, $78{\sim}100$ points). The average preoperative intermetatarsal angle was $14.4^{\circ}$, which was decreased to $7.9^{\circ}$ after the osteotomy with an average correction of $6.5^{\circ}$ and The average preoperative hallux valgus angle was $34.1^{\circ}$, which was decreased to $11.1^{\circ}$ after the osteotomy with an average correction of $23^{\circ}$. This new modified technique would prevent the angulation or shortening at the osteotomy site and it was also even more stable at osteotomy site, and could do even more effective and accurate correction of the deformity than conventional Chevron osteotomy. Conclusion: New modified chevron osteotomy for the treatment of symptomatic hallux valgus was done in 55 cases, and the results were satisfactory in all cases. This method was more stable at the osteotomy site than conventional Chevron osteotomy and was also possible to do more accurate and more effective correction of the deformity. It was also easy to control the distal fragment of first metatarsal bone.
Purpose: We try to retrospectively evaluated the amount of dorsal angulation angle of the first metatarsal commonly occurring as the complication of proximal dome osteotomy for hallux valgus. Materials and Methods: Between January 2004 and March 2004, 34 patients who underwent proximal dome osteotomy for moderate to severe hallux valgus. Two of 34 patients were male, and thirty-two were female. The average age was 57.6 years. We measured and compared hallux valgus angle, 1st-2nd intermetatarsal angle, dorsal angulation angle of 1st metatarsal on preoperative, postoperative, postoperative 3 weeks', postoperative 3 months' X-ray. Results: Osteotomy sites were completely united on plane X-ray in all cases. The hallux valgus angle averaged $41.2^{\circ}$ ($30{\sim}60^{\circ}$) at preoperative, $4.3^{\circ}$ ($-10{\sim}20^{\circ}$) at postoperative, $5.5^{\circ}$ ($-1{\sim}20^{\circ}$) at 3 weeks after operation, $7.8^{\circ}$ ($-2{\sim}20^{\circ}$) at 3 months after operation. The 1st-2nd intermetatarsal angle averaged $17.1^{\circ}$ ($12{\sim}24^{\circ}$) at preoperative, $6.3^{\circ}$ ($0{\sim}13^{\circ}$) at postoperative, $7.2^{\circ}$ ($0{\sim}15^{\circ}$) at 3 weeks after operation, $8.7^{\circ}$ ($0{\sim}18^{\circ}$) at 3 months after operation. The dorsal angulation angle averaged $0.4^{\circ}$ ($0{\sim}3^{\circ}$) at postoperative, $1.6^{\circ}$ ($0{\sim}7^{\circ}$) at 3 weeks after operation, $2.1^{\circ}$ ($0{\sim}8^{\circ}$) at 3 months after operation. There were no statistically correlation between increase of dorsal angulation angle of the distal segment of the first metatarsal and increase of hallux valgus angle or 1st-2nd intermetatarsal angle. Conclusion: Our results shows that the dorsal angulation of distal fragment occurring after the proximal dome osteotomy in the treatment of hallux valgus may be minimized with meticulous surgery and patient's education.
Objective : To evaluate the effect of calcium supplementation on spinal bone fusion in ovariectomized (OVX) rats. Methods : Sixteen female Sprague Dawley rats underwent bilateral ovariectomy at 12 weeks of age to induce osteoporosis and were randomly assigned to two groups : control group (n=8) and calcium-supplemented group (OVX-Ca, n=8). Autologous spinal bone fusion surgery was performed on both groups 8 weeks later. After fusion surgery, the OVX-Ca group was supplemented with calcium in drinking water for 8 weeks. Blood was obtained 4 and 8 weeks after fusion surgery. Eight weeks after fusion surgery, the rats were euthanized and the L4-5 spine removed. Bone fusion status and fusion volume were evaluated by manual palpation and three-dimensional computed tomography. Results : The mean fusion volume in the L4-5 spine was significantly greater in the OVX-Ca group ($71.80{\pm}8.06mm^3$) than in controls ($35.34{\pm}8.24mm^3$) (p<0.01). The level of osteocalcin, a bone formation marker, was higher in OVX-Ca rats than in controls 4 weeks ($610.08{\pm}10.41$ vs. $551.61{\pm}12.34$ ng/mL) and 8 weeks ($552.05{\pm}19.67$ vs. $502.98{\pm}22.76$ ng/mL) after fusion surgery (p<0.05). The level of C-terminal telopeptide fragment of type I collagen, a bone resorption marker, was significantly lower in OVX-Ca rats than in controls 4 weeks ($77.07{\pm}12.57$ vs. $101.75{\pm}7.20$ ng/mL) and 8 weeks ($69.58{\pm}2.45$ vs. $77.15{\pm}4.10$ ng/mL) after fusion surgery (p<0.05). A mechanical strength test showed that the L4-5 vertebrae in the OVX-Ca group withstood a 50% higher maximal load compared with the controls (p<0.01). Conclusion : Dietary calcium given to OVX rats after lumbar fusion surgery improved fusion volume and mechanical strength in an ovariectomized rat model.
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