Antioxidant Activities in Kenaf (Hibiscus cannabinus) Shoots during Growth Stages and Destination of Chlorogenic Acid and Kaempferol Glycosides.

Apart from being utilized as a commercial fiber at maturity, kenaf shoots have potential as a food and feed source because of their diverse bioactivities. Previous studies have focused on mature stems because of their high biomass, whereas the antioxidant activities (AA) and the destination of AA contributors of kenaf stems and their high-yielding byproduct leaves during the growth stage have rarely been studied. Therefore, we investigated changes in AA and its relative components in kenaf leaves and stems during the four vital growth stages. Higher ABTS radical cation and DPPH radical scavenging abilities and ferric reducing antioxidant power, total phenolic content, total flavonoid content, and total polysaccharide content were observed at all leaf stages and in the late stem stages. Chlorogenic acid (CGA) and kaempferol glycosides, especially kaempferitrin (Kfr), were identified as representative phenolic acids and flavonoids in both kenaf leaves and stems. The content of CGA in both leaves and stems increased corresponding to the plant's growth stage, whereas kaempferol glycosides were enhanced in leaves but declined in stems. The highest correlation was observed between TPC and AA in all organs. Further evaluation of CGA and Kfr verified that CGA was the predominant contributor to AA, surpassing Kfr. These findings suggest that kenaf leaves increase antioxidant levels as they grow and can be a useful source of stem harvesting byproducts.

Intraoperative fluoroscopy reduces the variability in femoral tunnel placement during single-bundle anterior cruciate ligament reconstruction.

PURPOSE: To evaluate the effect of using intraoperative fluoroscopy on femoral and tibial tunnel positioning variability in single-bundle anterior cruciate ligament (ACL) reconstruction. METHODS: A total of 80 consecutive patients with single-bundle ACL reconstruction between 2014 and 2016 were retrospectively reviewed. Among them, 40 underwent ACL reconstruction without fluoroscopy (non-fluoroscopy group) and 40 underwent fluoroscopy-assisted ACL reconstruction (fluoroscopy group). Femoral and tibial tunnel locations were evaluated using a standardized grid system with three-dimensional computed tomography images. Femoral and tibial tunnel location variability was compared between the groups. RESULTS: The operation time was longer in the fluoroscopy group than in the non-fluoroscopy group (61.3 ± 5.2 min vs. 55.5 ± 4.5 min, p < 0.001). In the fluoroscopy group, a guide pin was repositioned in 16 (40%) cases on the femoral side and 2 (5%) cases on the tibial side. No significant difference in the femoral tunnel location was observed between the fluoroscopy and non-fluoroscopy groups (anterior-posterior plane, 29.0% ± 3.2% vs. 30.0% ± 6.1%; proximal-distal plane, 30.8% ± 4.8% vs. 29.4% ± 8.3%; all parameters, n.s.); variability was significantly lower in the fluoroscopy group (p < 0.001 for both anterior-posterior and proximal-distal planes). No significant difference in the tibial tunnel location and variability was observed between the fluoroscopy and non-fluoroscopy groups (medial-lateral plane, 45.8% ± 2.0% vs. 46.6% ± 2.4%; anterior-posterior plane, 31.2% ± 4.0% vs. 31.0% ± 5.4%) (all parameters, n.s.). CONCLUSIONS: Tunnel positioning with fluoroscopic assistance is feasible and effective in achieving consistency in femoral tunnel placement despite a slightly longer operation time. Intraoperative fluoroscopy can be helpful in cases wherein identifying anatomical landmarks on arthroscopy was difficult or for surgeons with less experience who performed ACL reconstruction. LEVEL OF EVIDENCE: IV.

Second-look arthroscopic findings and clinical outcomes of meniscal repair with concomitant anterior cruciate ligament reconstruction: comparison of suture and meniscus fixation device.

INTRODUCTION: The purpose of this study was to evaluate the healing rate of repaired meniscus and functional outcomes of patients who received all-inside meniscal repair using sutures or devices with concomitant arthroscopic anterior cruciate ligament (ACL) reconstruction. MATERIALS AND METHODS: Among the patients who have ACL tear and posterior horn tear of medial or lateral meniscus, 61 knees who received all-inside repair using sutures (suture group, n = 28) or meniscal fixation devices (device group, n = 33) with concomitant ACL reconstruction during the period from January 2012 to December 2015, followed by second-look arthroscopy, were retrospectively reviewed. Healing status of the repair site was assessed by second-look arthroscopy. Through the clinical assessment, clinical success (negative medial joint line tenderness, no history of locking or recurrent effusion, and negative McMurray test) rate of the repaired meniscus and functional outcomes (International Knee Documentation Committee subjective score and Lysholm knee score) was evaluated. RESULTS: In a comparison of healing status of repaired meniscus evaluated by second-look arthroscopy, suture group had 23 cases of complete healing (82.1%), 4 cases of incomplete healing (14.3%), and 1 case of failure (3.6%). Device group had 18 cases of complete healing (54.5%), 4 cases of incomplete healing (24.2%), and 7 cases of failure (21.2%) (p = 0.048). Clinical success rate of the meniscal repair was 89.3% (25 cases) and 81.8% (27 cases) in suture group and device group, respectively (p = 0.488). No significant difference of functional outcomes was observed between the two groups (p > 0.05, both parameters). CONCLUSIONS: Among the patients who received meniscal repair with concomitant ACL reconstruction, suture group showed better healing status of repaired meniscus based on the second-look arthroscopy than device group. However, no significant between-group difference of clinical success rate and functional outcomes was observed.

Long-term outcomes of unicompartmental knee arthroplasty in patients requiring high flexion: an average 10-year follow-up study.

INTRODUCTION: To evaluate the long-term survival of unicompartmental knee arthroplasty (UKA) in the Asian population and assess differences in clinical outcomes between mobile- and fixed-bearing UKA. MATERIALS AND METHODS: Among 111 cases of UKA that were performed by 1 surgeon from January 2002 to December 2009, we retrospectively reviewed 96 cases (36 mobile-bearing, 62 fixed-bearing) for this study. We examined cause of revision or failure, type of reoperation/revision, and duration from the surgery date to the revision upon reviewing the medical record. Survival analysis was conducted using the Kaplan-Meier method. Functional outcomes were evaluated based on range of motion and patient-reported outcome (PRO) measures (Knee Injury and Osteoarthritis Outcome Score) for cases with at least 8 years of follow-up (average, 10.2 years). RESULTS: Overall, the 10-year survival was 88% [95% confidence interval (CI) 0.81-0.95], and the estimated mean survival time was 13.4 years (95% CI 12.5-14.2). In a comparison of survival between the mobile- and fixed-bearing groups, the former had a 10-year survival of 85% (95% CI, 0.72-0.97) and an estimated mean survival time of 13.5 years (95% CI 12.2-14.7) and the latter had a 10-year survival of 90% (95% CI 0.82-0.99) and an estimated mean survival time of 13.4 years (95% CI 12.3-14.4). Thus, there was no significant difference in survival between the two groups (log-rank test, p = 0.718). In addition, no significant difference in functional outcomes was observed between the two groups (p > 0.05 for all). CONCLUSIONS: UKA performed in the Asian population showed a relatively good functional outcome and survival rate at an average 10-year follow-up. No difference in survival and PROs was observed according to the bearing type. Although the present study demonstrated a good survival rate, similar to that in other Western studies, further studies investigating the impact of the Asian lifestyle on the long-term survival of UKA is necessary.

Multicast silicon photonic MEMS switches with gap-adjustable directional couplers.

We report on 4x20 silicon photonic MEMS switch that is capable of multicasting. The switch is built on passive optical crossbar network with gap-adjustable directional couplers. The switch has high on-off extinction ratio (59 dB), low insertion loss (< 4.0 dB), small footprint (1.2x4.5 mm2), and fast response (9.8 µs). The switching voltage is 9.6 V and 20 dB bandwidth is 31.5 nm. One-to-two and one-to-four multicast operations are demonstrated.

Non-fluorescent nanoscopic monitoring of a single trapped nanoparticle via nonlinear point sources.

Detection of single nanoparticles or molecules has often relied on fluorescent schemes. However, fluorescence detection approaches limit the range of investigable nanoparticles or molecules. Here, we propose and demonstrate a non-fluorescent nanoscopic trapping and monitoring platform that can trap a single sub-5-nm particle and monitor it with a pair of floating nonlinear point sources. The resonant photon funnelling into an extremely small volume of ~5 × 5 × 7 nm3 through the three-dimensionally tapered 5-nm-gap plasmonic nanoantenna enables the trapping of a 4-nm CdSe/ZnS quantum dot with low intensity of a 1560-nm continuous-wave laser, and the pumping of 1560-nm femtosecond laser pulses creates strong background-free second-harmonic point illumination sources at the two vertices of the nanoantenna. Under the stable trapping conditions, intermittent but intense nonlinear optical spikes are observed on top of the second-harmonic signal plateau, which is identified as the 3.0-Hz Kramers hopping of the quantum dot trapped in the 5-nm gap.

Size Regulation and Stability Enhancement of Pt Nanoparticle Catalyst via Polypyrrole Functionalization of Carbon-Nanotube-Supported Pt Tetranuclear Complex.

A novel multiwall carbon nanotube (MWCNT) and polypyrrole (PPy) composite was found to be useful for preparing durable Pt nanoparticle catalysts of highly regulated sizes. A new pyrene-functionalized Pt4 complex was attached to the MWCNT surface which was functionalized with PPy matrix to yield Pt4 complex/PPy/MWCNT composites without decomposition of the Pt4 complex units. The attached Pt4 complexes in the composite were transformed into Pt0 nanoparticles with sizes of 1.0-1.3 nm at a Pt loading range of 2 to 4 wt %. The Pt nanoparticles in the composites were found to be active and durable catalysts for the N-alkylation of aniline with benzyl alcohol. In particular, the Pt nanoparticles with PPy matrix exhibited high catalyst durability in up to four repetitions of the catalyst recycling experiment compared with nonsize-regulated Pt nanoparticles prepared without PPy matrix. These results demonstrate that the PPy matrix act to regulate the size of Pt nanoparticles, and the PPy matrix also offers stability for repeated usage for Pt nanoparticle catalysis.

Detection of Korean Native Honey and European Honey by Using Duplex Polymerase Chain Reaction and Immunochromatographic Assay.

Korean native honey (KNH) is much more expensive than European honey (EH) in Korea, because KNH is a favored honey which is produced less than EH. Food fraud of KNH has drawn attention of the government office concerned, which is in need of a method to differentiate between KNH and EH which are produced by the Asiatic honeybee, Apis cerana and the European honeybee, Apis mellifera, respectively. A method to discriminate KNH and EH was established by using duplex polymerase chain reaction (PCR) in this study. Immunochromatographic assay (IC) was examined to analyze the duplex PCR product. The DNA sequences of primers for the duplex PCR were determined by comparing cytochrome C oxidase genes of the two honey bee species. Chelex resin method was more efficient in extracting genomic DNA from honey than the other two procedures of commercial kits. The duplex PCR amplifying DNA of 133 bp were more sensitive than that amplifying DNA of 206 bp in detecting EH in the honey mixture of KNH and EH. Agarose gel electrophoresis and IC detected the DNA of 133 bp at the ratios of down to 1% and 5% EH in the honey mixture, respectively and also revealed that several KNH products distributed by internet shopping sites were actually EH. In conclusion, the duplex PCR with subsequent IC could also discriminate between KNH and EH and save time and labor.

Structural Analysis of Recombinant Human Preproinsulins by Structure Prediction, Molecular Dynamics, and Protein-Protein Docking.

More effective production of human insulin is important, because insulin is the main medication that is used to treat multiple types of diabetes and because many people are suffering from diabetes. The current system of insulin production is based on recombinant DNA technology, and the expression vector is composed of a preproinsulin sequence that is a fused form of an artificial leader peptide and the native proinsulin. It has been reported that the sequence of the leader peptide affects the production of insulin. To analyze how the leader peptide affects the maturation of insulin structurally, we adapted several in silico simulations using 13 artificial proinsulin sequences. Three-dimensional structures of models were predicted and compared. Although their sequences had few differences, the predicted structures were somewhat different. The structures were refined by molecular dynamics simulation, and the energy of each model was estimated. Then, protein-protein docking between the models and trypsin was carried out to compare how efficiently the protease could access the cleavage sites of the proinsulin models. The results showed some concordance with experimental results that have been reported; so, we expect our analysis will be used to predict the optimized sequence of artificial proinsulin for more effective production.

Recombinant Glargine Insulin Production Process Using Escherichia coli.

Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrix-assisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.

A nondestructive approach for discrimination of the origin of sesame seeds using ED-XRF and NIR spectrometry with chemometrics.

An energy dispersive X-ray fluorescence (ED-XRF) spectrometer and a near infrared (NIR) spectrometer combined with chemometrics were applied for origin discrimination of 48 Korean, 44 Chinese, and 21 Indian sesame seed samples used for development of a discriminant calibration model. Multi-elemental ED-XRF analysis based on Mg, Al, Si, P, S, Cl, K, Ca, Mn, Fe, and Cu was used for comparisons among origins. All elements, except for Fe, showed differences and 96.5% of seed samples were assigned to the correct origin using discriminant analysis based on chemical analytical results. NIR measurements were performed for spectral scanning. Classification of seeds using NIR discriminant analysis achieved 89.4% of seed samples assigned to the correct origin. Both ED-XRF and NIR are useful as nondestructive tools for discrimination of sesame seed origins.

Large-Scale Refolding and Enzyme Reaction of Human Preproinsulin for Production of Human Insulin.

Human insulin is composed of 21 amino acids of an A-chain and 30 amino acids of a B-chain. This is the protein hormone that has the role of blood sugar control. When the recombinant human proinsulin is expressed in Escherichia coli, a serious problem is the formation of an inclusion body. Therefore, the inclusion body must be denatured and refolded under chaotropic agents and suitable reductants. In this study, H27R-proinsulin was refolded from the denatured form with ß-mercaptoethanol and urea. The refolding reaction was completed after 15 h at 15°C, whereas the reaction at 25°C was faster than that at 15°C. The refolding yield at 15°C was 17% higher than that at 25°C. The refolding reaction could be carried out at a high protein concentration (2 g/l) using direct refolding without sulfonation. The most economical and optimal refolding condition for human preproinsulin was 1.5 g/l protein, 10 mM glycine buffer containing 0.6 M urea, pH 10.6, and 0.3 mM ß-mercaptoethanol at 15°C for 16 h. The maximum refolding yield was 74.8% at 15°C with 1.5 g/l protein. Moreover, the refolded preproinsulin could be converted into normal mature insulin with two enzymes. The average amount of human insulin was 138.2 g from 200 L of fermentation broth after enzyme reaction with H27R-proinsulin. The direct refolding process for H27R-proinsulin was successfully set up without sulfonation. The step yields for refolding and enzyme reaction were comparatively high. Therefore, our refolding process for production of recombinant insulin may be beneficial to the large-scale production of other biologically active proteins.

Identification of Porcine Endogenous Retrovirus (PERV) packaging sequence and development of PERV packaging viral vector system.

Studies of the retroviruses have focused on the specific interaction of the nucleocapsid protein with a packaging signal in the viral RNA as important for this selectivity, but the packaging signal in porcine endogenous retrovirus (PERV) has not been defined. Herein, we identified and analyzed this packaging signal in PERV and found hairpin structures with conserved tetranucleotides in their loops and nucleocapsid recognition sequences; both of which are key elements in the viral packaging signal of MLV. We evaluated packaging efficiency of sequence variants isolated from viral and proviral integrated genomes. All viral packaging sequences (Ψ) were identical, while five distinct packaging sequences were identified from proviral sources. One proviral sequence (Ψ1) was identical to that of the viral Ψ and had the highest packaging efficiency. Three variants (Ψ2, Ψ3, Ψ4) maintained key elements of the viral packaging signal, but had nucleotide replacements and consequently demonstrated reduced packaging efficiency. Despite of the same overall hairpin structure, the proviral variant (Ψ5) had only one GACG sequence in the hairpin loop and showed the lowest packaging efficiency other than ∆Ψ, in which the essential packaging sequence was removed. This result, thus, defined the packaging sequences in PERV and emphasized the importance of nucleotide sequence and RNA structure in the determination of packaging efficiency. In addition, we demonstrate efficient infection and gene expression from the PERV based viral vector, which may serve as a novel alternative to current retroviral expression systems.

Acupuncture for acute postoperative pain after back surgery: a systematic review and meta-analysis of randomized controlled trials.

OBJECTIVES: Acupuncture is commonly used as a complimentary treatment for pain management. However, there has been no systematic review summarizing the current evidence concerning the effectiveness of acupuncture for acute postoperative pain after back surgery. This systematic review aimed at evaluating the effectiveness of acupuncture treatment for acute postoperative pain (≤1 week) after back surgery. METHODS: We searched 15 electronic databases without language restrictions. Two reviewers independently assessed studies for eligibility and extracted data, outcomes, and risk of bias. Random effect meta-analyses and subgroup analyses were performed. RESULTS: Five trials, including 3 of high quality, met our inclusion criteria. The meta-analysis showed positive results for acupuncture treatment of pain after surgery in terms of the visual analogue scale (VAS) for pain intensity 24 hours after surgery, when compared to sham acupuncture (standard mean difference -0.67 (-1.04 to -0.31), P = 0.0003), whereas the other meta-analysis did not show a positive effect of acupuncture on 24-hour opiate demands when compared to sham acupuncture (standard mean difference -0.23 (-0.58 to 0.13), P = 0.21). CONCLUSION: Our systematic review finds encouraging but limited evidence for the effectiveness of acupuncture treatment for acute postoperative pain after back surgery. Further rigorously designed clinical trials are required.

Formaldehyde and heavy metal migration from rubber and metallic packaging/utensils in Korea.

The aim of this study was to determine the non-intentionally added substances--formaldehyde and trace metals--at 4% acetic acid conditions in rubber and metallic packaging/utensils. The temperature effect on migration in rubber and metallic packaging/utensils was monitored at 60 °C and 100 °C under acidic (pH < 3) circumstances. The concentrations were: formaldehyde--23.1 µg kg⁻¹, lead--13.41 µg kg⁻¹, cadmium--0.15 µg kg⁻¹, total arsenic--2.02 µg kg⁻¹ and nickel--2.92 µg kg⁻¹ at 60 °C and formaldehyde--148.9 µg kg⁻¹, lead--17.04 µg kg⁻¹, cadmium--0.14 µg kg⁻¹, total arsenic--7.25 µg kg⁻¹ and nickel--8.7 µg kg⁻¹ at 100 °C. A significant difference was noticed in formaldehyde and total arsenic between both temperatures (p < 0.01), which was not present in other trace metals. In conclusion, formaldehyde and total arsenic were more sensitive with cooking temperature than the other metals.

Primo Vessel Stressed by Lipopolysaccharide in Rabbits.

For tracking the primo vascular system, we observed the primo vessels in vivo in situ using the lipopolysaccharide (LPS) response in the lymphatic vessels of a rabbit. Injection of LPS (200 µg/kg) into the lymph nodes resulted in greatly stained primo vessels, which were swollen in some cases. We were able to obtain comparative images through alcian blue and diaminobenzidine staining, which clearly showed different morphologies of the primo vessels. The mechanism causing the response of the primo vessels to the injected LPS is still unclear; however, these results might be a first attempt at giving an explanation of the function of the primo vascular system and identifying the changes in the structure and function of the primo vascular system in response to an external stimulus such as an injection of LPS.

Enhanced Anti-inflammatory Effects of γ-irradiated Pig Placenta Extracts.

Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti-inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1ß, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1ß, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.

Effects of Yogurt Containing Fermented Pepper Juice on the Body Fat and Cholesterol Level in High Fat and High Cholesterol Diet Fed Rat.

This experiment investigated whether yogurt containing fermented pepper juice (FPJY) affects cholesterol level in high fat and high cholesterol diet (HFCD) fed rat. Twenty five Sprague-Dawley male rats of 7 wk were divided into 5 groups, and fed following diets for 9 wk; CON (control diet), HFCD (HFCD), PY (HFCD supplemented with 2% of plain yogurt), LFY (HFCD supplemented with 2% of FPJY), and HFY (HFCD supplemented with 5% of FPJY). In the LFY group, hepatic total lipid level decreased significantly compared to the HFCD group (p<0.05). Serum HDL cholesterol level tended to increase and hepatic total cholesterol level decreased and were comparable to the CON group (p>0.05). In HFY group, body weight and hepatic total lipid level significantly decreased over the HFCD group (p<0.05). Serum and hepatic total cholesterol level, kidney, and body fat weights decreased, and were compared to the CON group (p>0.05). Liver weight decreased as FPJY content was increased. Results suggested FPJY would inhibit organ hypertrophy and accumulation of body fat, hepatic lipid, and cholesterol in HFCD fed rat.

Expression and purification of recombinant human granulocyte colony-stimulating factor in fed-batch culture of Escherichia coli.

Granulocyte colony-stimulating factor (G-CSF) is a cytokine that has multiple roles in hematopoietic cells such as the regulation of proliferation and differentiation. Here, we describe fed-batch culture, refolding, and purification of rhG-CSF. The suitability of urea or sarcosine for solubilizing inclusion bodies (IBs) was tested. It was observed that urea is more efficient for solubilizing and refolding IBs than sarcosine is. The purity of rhG-CSF and the removal percentage of the rhG-CSF isoforms during purification were increased by pH 5.5 precipitation. The purity and the yield of purified rhG-CSF were 99% and 0.5 g of protein per liter culture broth, respectively. Our protocols of recombinant protein purification using ion exchange chromatography and semipreparative high performance liquid chromatography of pH-precipitated refolded solution may be informative to the industrial scale production of biopharmaceuticals.