rhBMP-2-Coated Acellular Dermal Graft for Chronic Rotator Cuff Healing: Translational Tendon Repair Research

A rotator cuff tear is a common shoulder injury in sports medicine. However, a rotator cuff repair still has the high failure rate (57%) in large torn (>8 cm2) rotator cuff cases. One of the main reasons is failing at suture-tendon cause of continuous tensional and torsional stresses even after surgery, and thus, an ideal biologic augmentation to overcome large tears is an essential challenge. The ECM graft, the biological material can be useful for augment repair of large torn rotator cuff. Recombinant human bone morphogenetic protein 2 (rhBMP-2), which belongs to transforming growth factor-β superfamily, is well known as an osteoinductive growth factor. It plays an important role in the development of bone and cartilage. rhBMP-2 also facilitates chemotaxis in the host tissue. In this study, rhBMP-2-coated acellular dermal graft, which is isolated from human cadaveric donor, was transplanted in the rabbit with the chronic rotator cuff injury. The radiologic image, histomorphometric, histologic image analyses, and tensile test were performed to evaluate the effectiveness. The results showed the enhancement of increased host cell infiltration, new bone formation, and tensile mechanical property. The rhBMP-2-coated acellular dermal graft will be promising for chronic rotator cuff healing

The Injectable rhBMP-2-containing Collagen Gel for Tendon Healing in a Rabbit Extra-Articular Bone Tunnel Model

This rabbit animal study has a hypothesis that the collagen gel, which is injectable easily, can be an effective carrier for recombinant human bone morphogenetic protein 2 (rhBMP-2) for the tendon healing in a bone tunnel. The cut upper long digital extensor tendon of each rabbit was inserted into the proximal tibia bone tunnel, and rhBMP-2 conjugated collagen gel was injected into the tendon-bone tunnel interface using a syringe. Biomechanical and histological performances were analyzed at 3 and 6 weeks after surgery. The collagen sol at room temperature was transformed to a gel at 37°C. The rhBMP-2 was slowly released from the collagen gel for more than 4 weeks. The in vivo experiment showed the enhanced new fibrocartilage and bone tissue formation at 6 weeks after injecting the rhBMP-2-containing collagen gel. The calcification and enthesis-like tissue were detected radiologically in the repaired tendon-bone junction. The viscous collagen gel-containing rhBMP-2 increased the fusion rate of the repaired tendon in the bone tunnel. This study showed that viscous collagen gel can be an effective carrier for rhBMP-2 for tendon healing in the bone tunnel. The rhBMP-2-containing collagen gel will be promising for tendon-bone interface healing in the future

Proizvodnja antikomplementnih egzopolisaharida submerznim uzgojem gljive Flammulina velutipes

Seven species of basidiomycetes have been investigated for anti-complementary activity in hot water extracts and ethanol soluble fractions. Since Flammulina velutipes had the most potent activity, culture conditions for its mycelial growth were optimized to increase the production efficiency of anti-complementary exopolysaccharides. The optimal medium composition was (in g/L): galactose 15, sodium nitrate 5, glutamic acid 3, KH2PO4 2.5 and MgSO4·7H2O 0.6. Optimal production of anti-complementary activity was achieved at pH=3.5–5.5 and 25 °C. With these optimal medium and culture conditions, mycelial dry mass was maximized at 3.17 mg/mL after 6 days of cultivation in a 5-liter stirred-tank bioreactor, without pH control. The anti-complementary activity of exopolysaccharides increased sharply after 4 days of cultivation, and showed a high level at 5–6 days of cultivation. A periodate-labile carbohydrate moiety played a leading role in the anti-complementary activity exhibited by exopolysaccharide produced from F. velutipes. Results of tests on the anti-complementary activity in the absence of Ca²+ and immunoelectrophoresis indicated that the mode of complement activation by exopolysaccharide from F. velutipes is via both the classical and alternative pathways and that the activation degree is almost the same in each pathway.Istražena je antikomplementna aktivnost spojeva ekstrahiranih vrućom vodom i etanolom iz sedam vrsta gljiva stapčara. Optimirani su uvjeti uzgoja micelija gljive s najvećom aktivnosti, Flammulina velutipes, radi povećanja proizvodnje antikomplementnih egzopolisaharida. Optimalni sastav podloge bio je (u g/L): galaktoza 15, natrijev nitrat 5, glutamična kiselina 3, KH2PO4 2,5 i MgSO4·7H2O 0,6. Optimalna proizvodnja postignuta je pri pH=3,5-5,5 i 25 ºC. Pri tim uvjetima proizvedena je maksimalna količina suhe tvari od 3,17 mg/L nakon 6 dana uzgoja u bioreaktoru s miješalicom zapremnine 5 L, bez kontrole pH-vrijednosti. Antikomplementna aktivnost egzopolisaharida naglo se povećala nakon 4 dana, te je bila visoka nakon 5-6 dana uzgoja. Šećerni je ostatak, podložan djelovanju perjodata, glavni razlog antikomplementne aktivnosti egzopolisaharida gljive F. velutipes. Rezultati testiranja takve aktivnosti u odsutnosti Ca²+ iona i imunoelektroforeza upućuju na to da se komplementi egzopolisaharida iz F. velutipes aktiviraju klasičnim i alternativnim putem te da je stupanj aktivacije skoro jednak za oba puta

High-Throughput Screening of Acyl-CoA Thioesterase I Mutants Using a Fluid Array Platform

Screening target microorganisms from a mutated recombinant library plays a crucial role in advancing synthetic biology and metabolic engineering. However, conventional screening tools have several limitations regarding throughput, cost, and labor. Here, we used the fluid array platform to conduct high-throughput screening (HTS) that identified Escherichia coli ???TesA thioesterase mutants producing elevated yields of free fatty acids (FFAs) from a large (106) mutant library. A growth-based screening method using a TetA-RFP fusion sensing mechanism and a reporter-based screening method using high-level FFA producing mutants were employed to identify these mutants via HTS. The platform was able to cover >95% of the mutation library, and it screened target cells from many arrays of the fluid array platform so that a post-analysis could be conducted by gas chromatography. The ???TesA mutation of each isolated mutant showing improved FFA production in E. coli was characterized, and its enhanced FFA production capability was confirmed

Analysis of import changes through shift-share, location quotient and BCG techniques: Gwangyang Port in Asia

The main aim of this article is to analyze the import changes of Gwangyang Port using shift-share, location quotient and BCG matrix techniques. We perform the standard shift-share analysis and spatial shift-share analysis for the period 2010–2018 and investigate the import performance of Gwangyang Port for coal, iron ore, natural gas and vegetable matter. The static analysis shows that the regional shift effect, which is the most important component, is negative for coal and iron ore, but positive for natural gas and vegetable matter. The spatial shift-share analysis also indicates that Gwangyang Port experiences not only the gains in regional competitiveness but the industrial advantage for iron ore, natural gas and vegetable matter owing to its higher competitiveness. Incorporating location coefficients into BCG matrix for coal imports, we also show that Gwangyang Port succeeds upgrading its position for natural gas and vegetable matter, but fails escaping from transformation category or upgrading its position for coal and iron ore

Preparative Synthesis of dTDP-L-Rhamnose Through Combined Enzymatic Pathways

dTDP-L-rhamnose, an important precursor of O-antigen, was prepared on a large scale from dTMP by executing an one-pot reaction in which six enzymes are involved. Two enzymes, dTDP-4-keto-6-deoxy-D-glucose 3,5-epimerase and dTDP-4-keto-rhamnose reductase, responsible for the conversion of dTDP-4-keto-6-deoxy- D-glucose to dTDP-L-rhamnose, were isolated from their putative sequences in the genome of Mesorhizobium loti, functionally expressed in Escherichia coli, and their enzymatic activities were identified. The two enzymes were combined with an enzymatic process for dTDP-4- keto-6-deoxy-D-glucose involving TMP kinase, acetate kinase, dTDP-glucose synthase, and dTDP-glucose 4,6- dehydratase, which allowed us to achieve a preparative scale synthesis of dTDP-L-rhamnose using dTMP and glucose-1-phosphate as starting materials. About 82% yield of dTDP-L-rhamnose was obtained based on initial dTMP concentration at 20 mM dTMP, 1 mM ATP, 10 mM NADH, 60 mM acetyl phosphate, and 80 mM glucose-1- phosphate. From the reaction with 20 ml volume, approximately 180 mg of dTDP-L-rhamnose was obtained in an overall yield of 60% after two-step purification, that is, anion exchange chromatography and gel filtration for desalting. The purified product was identifiedbyHPLC, ESI-MS,andNMR,showingabout95%purity

Effect of Apneic Oxygenation via Nasal Prong on Gas Exchange and Vital Signs during Fiberoptic Intubation under General Anesthesia

A clinical study for the evaluation of the effect of apneic oxygenation by nasal prong during fiberoptic orotracheal intubation on gas exchange and vital signs has been done on the patients who received tympanomastoidectomy (ASA c1assfication 1 and 2, aged from 20 to 40). Among them, 22 patients were selected whose fiberoptic intubation lasted more than 3 but less than 4 minutes, to observe the changes of Pa02, PaC02, HR, and MAP. 11 patients who underwent fiberoptic orotracheal intubation in apneic state without oxygen administration (Group I) showed similar increases in vital signs to the other 11 patients who received apneic oxygenation (Group II). PaC02 increased more in Group I than in Group II, which was not statistically significant. The differences of Pa02 at 1 and 2 minutes between two groups after removal of oxygen mask and beginning of fiberoptic intubation, were not statistically significant but Group II showed a significantly lesser degree of decrease in Pa02 at 3 minutes. We might say that apneic oxygenation during fiberoptic intubation under general anesthesia is useful because it could delay the onset of hypoxia, thereby provide extra time for intubation. Therefore we could attempt intubation up to 3 minutes on the fully relaxed patient, if we give oxygen via nasal prong