In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.
Background: There is an opinion that improper postures of the head and cervical spine are associated with temporomandibular joint (TMJ) disorders (TMDs). Objects: The aim of this study was to investigate the proportions among the cervical kyphotic angle, physical symptoms including the pain intensity level of the TMJ, and severity of TMD disability in patients diagnosed with TMD. Methods: Sixty-two subjects participated in the study. The evaluation tools included measurements of the cervical kyphotic angle based on the Ishihara index, pressure pain threshold (PPT) on the TMJ, maximal mouth opening (MMO) without pain, current pain intensity level of the TMJ measured using the Quadruple Visual Analogue Scale (QVAS), Korean TMD (KTMD) disability index, KTMD Symptom Frequency/Intensity Scales (SFS/SIS), and Korean Headache Impact Test-6. Correlation analysis was conducted to investigate the correlations between the cervical kyphotic angle and parameters related to TMJ symptoms. Results: Variables that were significantly correlated with the cervical kyphotic angle were the PPT around the TMJ (r = 0.259, p < 0.05), current pain intensity level of the TMJ based on the QVAS (r = -0.601, p< 0.01), and usual pain intensity level based on the SIS (r = -0.379, p < 0.01). The level of TMD functional disability was significantly correlated with the degree of headache (r = 0.551, p < 0.01), level of PPT of the TMJ (r = -0.383, p < 0.01), pain-free MMO (r = -0.515, p < 0.01), pain intensity level of the TMJ based on the QVAS (r = 0.393, p < 0.01), TMD symptom frequency (r = 0.739, p < 0.01), usual pain intensity of the TMJ (r = 0.624, p < 0.01), and most severe pain intensity of the TMJ (r = 0.757, p < 0.01). Conclusion: There is a positive correlation between the cervical kyphotic angle and PPT and a negative correlation between the current and usual pain intensity levels of the TMJ. The cervical kyphotic angle was a predictor of the pain level, tenderness threshold, and intensity of pain in the TMJ.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.9
/
pp.1347-1355
/
2015
In this study, the physicochemical properties, antioxidant capacities, and volatile flavor compounds of oriental melon wine prepared by freeze concentration after heat treatment (HA), ascorbic acid treatment (AAT), and heat and ascorbic acid treatment (HAAT) were investigated. During fermentation period, the melon wine by HAAT showed greater reduction of soluble solids and reducing sugar contents compared to other treatments. In addition, the melon wine treated with HAAT also showed a higher L value and lower browning index compared to other treatments. After aging, free sugar including fructose, and organic acids including citric acid, succinic acid, and malic acid were detected in all samples. For antioxidant activities and contents, HAAT treated wine showed greater antioxidant activities and total phenolic contents than those of others. In GC/MS analysis, a total of 33 volatile flavor compounds were identified. In the principal component analysis of volatile flavor compounds, principal components 1 and 2 represented 88.15% of the whole date distribution and showed opposite tendencies. Taken together, HAAT enhanced the antioxidant activities and sensory properties of oriental melon wine. Moreover, freeze concentration gave the different volatile flavor characteristics in oriental melon wine.
The purpose of this article to evaluate the availability of the rectus femoris flap in Korean subjects. Material and Methods is that Cadaveric dissections were done on 51 femoral triangles of 26 cadevers. We measured the length of the direct head of rectus femoris from anterior superior iliac spine to patella upper pole, ASIS to lateral border of femoral nerve, and entry point of femoral nerve and vessel branches to rectus. Usually, there were three terminal branches to rectus femoris from the femoral nerve. The entry point of the first branch was at the proximal $17.5{\sim}31.4%$ portion of the rectus femoris. The second and the third branch entered at the proximal $22.5{\sim}40.7%$ and $26.3{\sim}42.3%$, respectively. The vessel entry was at $20.2{\sim}37.3%$. The length from ASIS to femoral nerve was $3.5{\sim}8.5\;cm$. Among the 51 rectus femoris muscles, 44 had one nutrient artery, and 7 had 2 nutrient arteries. The nutrient artery originated from the descending branch of the lateral femoral circumflex artery in 18(40.9%) cases, directly from the lateral femoral circumflex artery in 8(18.0%) cases, and from proximal(6 cases, 13.6%) and distal(12 cases, 27.3%) portion of the deep femoral artery. The average length of the nutrient artery was 29.8 mm and the width was 2.14 mm. The point where it meets the main feeding artery of the rectus femoris was $9.0{\sim}15.0\;cm$ from the ASIS. In all cases, the main artery's entrance was proximal to the first nerve branch. Conclusion is that rectus femoris has available data for functional flap.
Journal of the Korea Academia-Industrial cooperation Society
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v.18
no.2
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pp.583-589
/
2017
The 1900s in New Orleans, the harbor city, was indeed an era of confusion because there were various ethnic groups and races. Songs that had been sung by slaves taken from Africa, Black spiritual music, blues, British folk songs, French folk music, ballet music, Spanish dance music, and the march of military bands were mixed with Rag Time to achieve diversity. This developed the beginning of jazz. While swing jazz was most popular and loved by the public during the 20th century, the bebop preferred the small scale organization of musical instruments and developed as a form of jazz featuring the impromptu musical performances. Later, cool jazz, a new style involving the fast and complicated code progress, emerged with free jazz, which features the fundamental rupture from the tradition of the jazz. Miles Davis, who introduced the rock beat in jazz, started fusion jazz. Although jazz has been named differently depending on the era, the main attraction of jazz lies on improvisation. In other words, despite a small changes in code progress and rhythm, the most important thing the player considers is improvisation. Some famous players who lived in the same era followed the whole atmosphere but each had their own style. So, even when they did play the same song, they revealed their style in solo parts despite the same head.
The occurrence and time course of capacitation, acrosomal loss, and hyperactivated motility require quantitative definition in order to characterize fertile human sperm. Recently the method has been developed to estimate the quality of spermatozoa by using kinematic parameters such as curvilinear velocity(VCL), average path velocity(VAP), linearity(LIN), straightness(STR), amplitude of lateral head displacement(ALH), and beat cross frequence(BCF) from Computer Assisted Sperm Analysis (CASA). In this study, using the Hamilton Thorn motility analyzer HTM 2030(Hamilton Thorn Research, Beverly, MA), we attempted to identify the spermatozoa with hyperactivated motility (HA) objectively and to monitor hyperactivation of human spermatozoa during incubation in capacitating media and after treatment of calcium ionophore as compared with acrosome status. And we examined whether HA are related to the result of SPA. Semen samples obtained from 16 healthy men were prepared by swim up technique and preincubated in a capacitating media(modified BWW medium) for 5 hours and treated with calcium ionophore solution. The acrosome reaction was detected with PSA-FITC labelling of the acrosome and in vitro sperm ferilizing capacity was assessed by the zona free hamster ovum penetration assay (SPA). The incidence of hyperactivated sperm was 2.6% in fresh semen, 14.3% of the swim up population, 13.7% after 5h of incubation. Significant increase of percentage of hyperactivated sperm was observed after the incubation (p<0.05) but after treatment, no significant changes of percentage of hyperactivated sperm(l1.8%) in contrast to significant rise in the percentage of acrosome reacted cells. Correlation analysis failed to show any significant relationship between the percentage of sperm with HA and SPA score. In conclusion, although no direct correlations were found between the results of SPA and HA, hyperactivation of sperm is associated with capacitation and monitoring hyperactivated sperm will be expected as a method of evaluating the functional quality of sperm such as SPA.
Kim, Il-Man;Lee, Chang Young;Son, Eun Ik;Kim, Dong Won;Yim, Man Bin;Kim, Sang Pyo
Journal of Korean Neurosurgical Society
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v.30
no.3
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pp.334-341
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2001
Objective : We have currently changed treatment strategies to methotrexate(MTX)-based preirradiation chemotherapy with subsequent planned radiation for the initial therapy of primary central nervous system lymphoma (PCNSL). The aim of this study was to evaluate the results of treating PCNSL with chemotherapy plus radiotherapy (CRT) or radiotherapy(RT) alone. Method and Material : This study involved 10 females and 3 males patients with a mean age of 54.2 years. All patients underwent surgery, open(8 cases) or stereotactic biopsy(5 cases) for histological diagnosis. Eleven tumors were diffuse large B-cell lymphomas. Tumor volume change in the follow-up images and survival time were evaluated in patients treated with CRT and RT alone. In the beginning, two patients received ProMACE-Cytabom chemotherapeutic regimen, but did not complete the course and died of progressive tumor 8 and 9 months after diagnosis, respectively. One patient died at 6 months before chemotherapy. These three were excluded from the survival analysis. Five patients(RT group) completed full courses of cranial irradiation with or without boost. For the current combined modality treatment, high-dose MTXbased chemotherapy(systemic and intrathecal MTX, IV vincristine, and oral procarbazine) followed by whole brain irrdiation to 45Gy to tumor was introduced in 5 patients of CRT group. Result : A complete response was achieved in three of five who received RT only and in all of five who received CRT. All patients in CRT groups are in disease free status at a mean 23 months following therapy. The RT group patients refused any additional salvage therapy at tumor relapse and survived at mean 20 months from diagnosis. The Karnofsky performance status improved in eight of ten patients with treatment. The treatment toxicity included leukoencephalopathy in RT group and severe leukopenia, transient hepatitis, avascular necrosis of femoral head, hearing loss, and amenorrhea in CRT group, respectively. Conclusion : The combined modality therapy of MTX-based chemotherapy plus radiotherapy for PCNSL may enhance tumor response and improve patient survival. The patients who received CRT should be carefully followed up because of the higher risk of treatment-induced late neurotoxicity.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.26
no.5
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pp.455-461
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2000
Malignant tumors of the head and neck frequently require treatment with both radiotherapy and surgery. Reconstruction of the defect in previously irradiated field is a challenge to surgeon, who must produce both a functional and an esthetic result. Hyperbaric oxygen therapy(HBO) has been used in an attempt to reduce the deleterious effects of radiation. But the issue of whether prior irradiation and HBO of the recipient site of a free flap affects the result of reconstruction continues to generate controversy. So, the effects of irradiation and hypergbaric oxygen therapy on microvascular anastomosis was evaluated in an experimental study in femoral vessels of rats. The experimental groups were divided into 3 groups, contorol group, irradiation group, and irradiation and HBO group. Preoperative irradiation was delivered in the left groin field with single dose corresponding 2,000cGy and total 48 hours of HBO was given 100% oxygen at 2.4 atmosphere for 4 weeks. The femoral vessels of 60 rats were anastomosed after irradiation and HBO treatment. Three days, 1 week, 2 weeks and 4 weeks after surgery, the femoral vessels were evaluated for patency and histopathologic changes. There was no notable effect of irradiation on patency of femoral vessels in rats and the radiation effects were obvious on histological examination which showed the sloughing of the endothelial cells, subintimal hyperplasia and fibrosis on the media and adventitia of femoral arteries. The histologic changes of the femoral veins were mild and not typical. But the effects of hyperbaric oxygen therapy after irradiation was seen not marked difference in irradiation group.
Tecce, Michael G.;Othman, Sammy;Mauch, Jaclyn T.;Nathan, Shelby;Tilahun, Estifanos;Broach, Robyn B.;Azoury, Said C.;Kovach, Stephen J.
Archives of Craniofacial Surgery
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v.21
no.4
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pp.229-236
/
2020
Background: Oncologic resection of the scalp confers several obstacles to the reconstructive surgeon dependent upon patient-specific and wound-specific factors. We aim to describe our experiences with various reconstructive methods, and delineate risk factors for coverage failure and complications in the setting of scalp reconstruction. Methods: A retrospective chart review was conducted, examining patients who underwent resection of fungating scalp tumors with subsequent soft-tissue reconstruction from 2003 to 2019. Patient demographics, wound and oncologic characteristics, treatment modalities, and outcomes were recorded and analyzed. Results: A total of 189 patients were appropriate for inclusion, undergoing a range of reconstructive methods from skin grafting to free flaps. Thirty-three patients (17.5%) underwent preoperative radiation. In all, 48 patients (25.4%) suffered wound site complications, 25 (13.2%) underwent reoperation, and 47 (24.9%) suffered from mortality. Preoperative radiation therapy was an independent risk factor for wound complications (odds ratio [OR], 2.85; 95% confidence interval [CI], 1.1-7.3; p=0.028) and reoperations (OR, 4.45; 95% CI, 1.5-13.2; p=0.007). Similarly, the presence of an underlying titanium mesh was an independent predictor of wound complications (OR, 2.49; 95% CI, 1.1-5.6; p=0.029) and reoperations (OR, 3.40; 95% CI, 1.2-9.7; p=0.020). Both immunosuppressed status (OR, 2.88; 95% CI, 1.2-7.1; p=0.021) and preoperative radiation therapy (OR, 3.34; 95% CI, 1.2-9.7; p=0.022) were risk factors for mortality. Conclusion: Both preoperative radiation and the presence of underlying titanium mesh are independent risk factors for wound site complications and increased reoperation rates following oncologic resection and reconstruction of the scalp. Additionally, preoperative radiation, along with an immunosuppressed state, may predict patient mortality following scalp resection and reconstruction.
Block, Kai Tobias;Chandarana, Hersh;Milla, Sarah;Bruno, Mary;Mulholland, Tom;Fatterpekar, Girish;Hagiwara, Mari;Grimm, Robert;Geppert, Christian;Kiefer, Berthold;Sodickson, Daniel K.
Investigative Magnetic Resonance Imaging
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v.18
no.2
/
pp.87-106
/
2014
Purpose : To describe how a robust implementation of a radial 3D gradient-echo sequence with stack-of-stars sampling can be achieved, to review the imaging properties of radial acquisitions, and to share the experience from more than 5000 clinical patient scans. Materials and Methods: A radial stack-of-stars sequence was implemented and installed on 9 clinical MR systems operating at 1.5 and 3 Tesla. Protocols were designed for various applications in which motion artifacts frequently pose a problem with conventional Cartesian techniques. Radial scans were added to routine examinations without selection of specific patient cohorts. Results: Radial acquisitions show significantly lower sensitivity to motion and allow examinations during free breathing. Elimination of breath-holding reduces failure rates for non-compliant patients and enables imaging at higher resolution. Residual artifacts appear as streaks, which are easy to identify and rarely obscure diagnostic information. The improved robustness comes at the expense of longer scan durations, the requirement for fat suppression, and the nonexistence of a time-to-center value. Care needs to be taken during the configuration of receive coils. Conclusion: Routine clinical use of radial stack-of-stars sequences is feasible with current MR systems and may serve as substitute for conventional fat-suppressed T1-weighted protocols in applications where motion is likely to degrade the image quality.
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