UPLC-QTOF-MS/MS screening and identification of bioactive compounds in fresh, aged, and browned Magnolia denudata flower extracts.

The Magnolia denudata flower is used to prepare tea and is often fermented to improve its flavor. Herein, fresh, aged, and browned M. denudata flower extracts were characterized using ultra performance liquid chromatography coupled with a hybrid quadrupole orthogonal time-of-flight mass spectrometer (UPLC-Q-TOF/MS/MS). The 1223 and 458 mass ions of ESI+ and ESI- modes that were significantly changed by the fermentation process were selected using three criteria. Sixteen compounds including flavonoids, phenylethanoid glycoside derivatives (PhGs), caffeoylquinic acids (CQAs), and others were identified based on their accurate mass and MS/MS spectra and analyzed as the main chemical components. The levels of the main chemical components changed after fermentation. The comparative quantity and composition of the phytochemicals differed for the three extract types. For example, flavonols were affected by fermentation, resulting in an increase or decrease (fold change value of negative ion: rutin -0.47; keioside 1.00). The CQAs were low during fermentation (1-CQA, -1.62; chlorogenic acid, -1.48). However, the quinic acid content was significantly high (quinic acid, 1.36). Isomers of PhGs like isoverbasoside and isoacteoside were produced during fermentation (isoverbasoside, 5.42; isoacteoside, B 3.33). These observations may provide a basis for studying the physiological effects of non-fermented and fermented M. denudata flower.

Structural and kinetic insights into flavin-containing monooxygenase and calponin-homology domains in human MICAL3.

MICAL is an oxidoreductase that participates in cytoskeleton reorganization via actin disassembly in the presence of NADPH. Although three MICALs (MICAL1, MICAL2 and MICAL3) have been identified in mammals, only the structure of mouse MICAL1 has been reported. Here, the first crystal structure of human MICAL3, which contains the flavin-containing monooxygenase (FMO) and calponin-homology (CH) domains, is reported. MICAL3 has an FAD/NADP-binding Rossmann-fold domain for mono-oxygenase activity like MICAL1. The FMO and CH domains of both MICAL3 and MICAL1 are highly similar in structure, but superimposition of the two structures shows a different relative position of the CH domain in the asymmetric unit. Based on kinetic analyses, the catalytic efficiency of MICAL3 dramatically increased on adding F-actin only when the CH domain was available. However, this did not occur when two residues, Glu213 and Arg530, were mutated in the FMO and CH domains, respectively. Overall, MICAL3 is structurally highly similar to MICAL1, which suggests that they may adopt the same catalytic mechanism, but the difference in the relative position of the CH domain produces a difference in F-actin substrate specificity.

Sulfate assimilation regulates hydrogen sulfide production independent of lifespan and reactive oxygen species under methionine restriction condition in yeast.

Endogenously produced hydrogen sulfide was proposed to be an underlying mechanism of lifespan extension via methionine restriction. However, hydrogen sulfide regulation and its beneficial effects via methionine restriction remain elusive. Here, we identified the genes required to increase hydrogen sulfide production under methionine restriction condition using genome-wide high-throughput screening in yeast strains with single-gene deletions. Sulfate assimilation-related genes, such as MET1, MET3, MET5, and MET10, were found to be particularly crucial for hydrogen sulfide production. Interestingly, methionine restriction failed to increase hydrogen sulfide production in mutant strains; however, it successfully extended chronological lifespan and reduced reactive oxygen species levels. Altogether, our observations suggested that increased hydrogen sulfide production via methionine restriction is not the mechanism underlying extended yeast lifespan, even though increased hydrogen sulfide production occurred simultaneously with yeast lifespan extension under methionine restriction condition.

Poly-γ-glutamic acid enhances the growth and viability of Chinese hamster ovary cells in serum-free medium.

The protective effects of polymer additives, including a group of viscosity-enhancing polymer poly-γ-glutamic acid (γPGA; 10, 50, and 500 kDa) and surface-active polymer Pluronic F68, on Chinese hamster ovary cells against damage due to shear stress were investigated in shake-flask cultures. The level of protection was dependent upon the molecular weight of γPGA and its concentration. When 0.05 or 0.075 % of 500 kDa γPGA was added, the cell growth and viability were almost equal to those of Pluronic F68 supplementation and were much higher than those of the control without additives. For the first time, we show that γPGA is another environmentally-friendly medium additive that can be used in place of Pluronic F68.

Simultaneous correction of hard- and soft-tissue facial asymmetry: combination of orthognathic surgery and face lift using a resorbable fixation device.

The purpose of this study was to evaluate outcomes of simultaneous correction of the hard- and soft-tissue facial asymmetry with face lift procedure using a resorbable fixation device (Endotine Ribbon; Coapt Systems, Palo Alto, CA) during bimaxillary orthognathic surgery in cases with severe facial asymmetry. The samples consisted of 8 patients (mean age, 23.3 [SD, 4.4] years; 8 skeletal class III and 2 class II malocclusion) who received bimaxillary orthognathic surgery and a face lift procedure using a resorbable fixation device. Preoperative cephalometric evaluation of the maxillary occlusal plane cant and chin point deviation and data on surgical movement, site, time, and difficulty of face lift procedure were collected at 1 week before operation and during operation procedure. The amounts of lip cant between preoperation and postoperation were compared. Pain, stability of fixation, adverse effects, relapse, and patients' and surgeon's satisfaction were evaluated at 6 months after operation. Initial and final amounts of the lip cant were 4.15 (SD, 0.53) and 0.80 (SD, 0.48) mm (correction rate, 76.8%). The face lift procedure took 28.4 (SD, 3.3) minutes without difficulty. There were no severe complications such as hematoma, facial nerve injury, and postoperative scar. By the patients' and surgeon's view, all had satisfactory jowl elevation, lip canting correction, and achievement of the soft-tissue symmetry without evidence of recurrent asymmetry or loss of fixation. If the face lift procedure using a resorbable fixation device is done with proper vector control during orthognathic surgery, the hard- and soft-tissue facial asymmetry can be corrected simultaneously with satisfactory outcomes.

Subnasale flap for correction of columella base deviation in secondary unilateral cleft lip nasal deformity.

OBJECTIVE: To present the effects of subnasale (SN) flap on the correction of deviated columella base in patients with secondary unilateral cleft lip nasal (UCLN) deformity. PATIENTS AND METHODS: The patients were 6 adult Koreans with cleft cases (mean age, 29.1 y; range, 20-43 y; 1 man and 5 women; secondary UCLN deformity with columella base deviation). After the open rhinoplasty with the Padovan incision had been completed for correction of the secondary UCLN deformity, the SN flap was used for correction of the deviated columella base. The design of the SN flap started from the central portion of the SN region that will be the reconstructed columella and extended to the nasal floor in the larger nostril site. Then, the SN flap was transposed into the smaller nostril site. The columella skin was also elevated and placed into the larger nostril site with the distal end inserting into the donor site. The patients' follow-up period was 16 to 40 months. RESULTS: The SN flap after the open rhinoplasty with the Padovan incision resulted in minimal scarring on the donor and recipient sites. The deviated columella base was straightened in all cases. During the surgical follow-up period, there was no recurrence of the columella base deviation in any patient and no other noteworthy complications were found. CONCLUSIONS: For correction of the columella base deviation in patients with secondary UCLN deformity, the SN flap can be a viable treatment option.

Validated robotic laparoscopic surgical training in a virtual-reality environment.

BACKGROUND: A robotic virtual-reality (VR) simulator has been developed to improve robot-assisted training for laparoscopic surgery and to enhance surgical performance in laparoscopic skills. The simulated VR training environment provides an effective approach to evaluate and improve surgical performance. This study presents our findings of the VR training environment for robotic laparoscopy. METHODS: Eight volunteers performed two inanimate tasks in both the VR and the actual training environment. The tasks were bimanual carrying (BC) and needle passing (NP). For the BC task, the volunteers simultaneously transferred two plastic pieces in opposite directions five times consecutively. The same volunteers passed a surgical needle through six pairs of holes in the NP task. Both tasks require significant bimanual coordination that mimics actual laparoscopic skills. Data analysis included time to task completion, speed and distance traveled of the instrument tip, as well as range of motion of the subject's wrist and elbow of the right arm. Electromyography of the right wrist flexor and extensor were also analyzed. Paired t-tests and Pearson's r were used to explore the differences and correlations between the two environments. RESULTS: There were no significant differences between the actual and the simulated VR environment with respect to the BC task, while there were significant differences in almost all dependent parameters for the NP task. Moderate to high correlations for most dependent parameters were revealed for both tasks. CONCLUSIONS: Our data shows that the VR environment adequately simulated the BC task. The significant differences found for the NP task may be attributed to an oversimplification in the VR environment. However, they do point to the need for improvements in the complexity of our VR simulation. Further research work is needed to develop effective and reliable VR environments for robotic laparoscopic training.

Use of soybean protein hydrolysates for promoting proliferation of human keratinocytes in serum-free medium.

Human keratinocytes are generally cultured in media containing bovine pituitary extract (BPE), an animal product that can be a source of infectious contaminants. We investigated whether a safer plant product could replace BPE in the culture medium. Medium containing both BPE and soy protein hydrolysates (Bacto Soytone and Soy Hydrolysate) produced the largest number of viable cells, followed in descending order by medium supplemented only with BPE, only with the hydrolysates, and without supplementation (basal medium only). Soybean protein is thus an excellent source of nutrients for the growth of adherent keratinocytes, although they do not fully substitute for BPE.

Short esophagus: how much length can we get?

INTRODUCTION: Laparoscopic antireflux surgery requires an adequate length of intra-abdominal esophagus. Short esophagus can cause wrap herniation and poor clinical outcomes. The aim of the study is to measure maximum length of esophageal elongation with transhiatal mediastinal dissection. METHODS: This is a review of a prospective database created in the tertiary referral center between 2003 and 2006. One hundred and six patients with gastroesophageal reflux disease and suspected short esophagus on barium swallow were studied. Patients underwent antireflux surgery with extended transhiatal mediastinal dissection to elongate short esophagus. Routine measurement of intra-abdominal esophageal segment length with intraoperative esophagogastroscopy and laparoscopy was utilized to define the gastroesophageal junction (GEJ) in order to quantify total intra-abdominal esophageal length. Postoperative 24-h pH manometry, UGI series, and symptom scores were recorded to document the clinical outcomes. The aim of the dissection was to mobilize>or=3 cm of intra-abdominal esophagus. RESULTS: Total esophageal elongation was achieved with a mean of 2.65 (range 2-18) cm. Resultant intra-abdominal esophageal length was measured with a mean of 3.15 (range of 3 to 5) cm. None of the preoperative "short esophagus" required Collis' gastroplasty post extended mediastinal dissection. All preoperative symptom scores showed significant improvements with mean follow-up of 18 (9-36) months. Mean distal esophageal acid exposure normalized in all patients studied postoperatively. CONCLUSION: Short esophagus can be safely elongated with extended mediastinal esophageal dissection. This technique can obviate the need for Collis' gastroplasty and improve overall outcome after antireflux surgery. We recommend that extended transhiatal mediastinal dissection be performed to establish 3 cm of intra-abdominal esophagus at the time of antireflux procedures.

Long-term outcome of cruroplasty reinforcement with human acellular dermal matrix in large paraesophageal hiatal hernia.

BACKGROUND: Laparoscopic repair of a large hiatal hernia using simple sutures only for the cruroplasty is associated with a high recurrence rate. The solution was to place synthetic mesh over the cruroplasty thereby decreasing recurrence rates in exchange for complications, such as gastric and esophageal erosions. Our initial report investigated the use of human acellular dermal matrix (AlloDerm) as a more suitable alternative. This study highlights our long-term results>1 year of cruroplasty reinforcement with AlloDerm in the repair of large hiatal hernias. METHODS AND MATERIAL: This is a retrospective study performed at our university. Between 2005 and 2006, 52 consecutive patients with large hiatal hernias had the cruroplasty site reinforced with AlloDerm. The variables analyzed were age, sex, weight, height, hiatal hernia size, operative time, length of hospital stay, follow-up, and postoperative complications. RESULTS: The mean for age was 56.7 years, for weight was 87.9 kg, for height 117 cm, for hernia size was 5.75 cm, operative time was 121 min, and for hospital stay was 1.36 days. Complication included pneumothorax, 3 (5.5%); atelectasis, 1 (1.9%); urinary retention, 1 (1.9%); and recurrence, 2 (3.8%). CONCLUSION: Laparoscopic hiatal hernia repair with reinforcement using human acellular dermal matrix can be performed safely with a short hospital stay and low rate of complications, especially a low rate of recurrence.

One hundred consecutive laparoscopic Nissen's without the use of a bougie.

BACKGROUND: The creation of a floppy and symmetric fundoplication over a bougie has been the standard of care in laparoscopic surgery. The use of a bougie carries a risk of esophageal perforation but lowers the risk of postoperative dysphagia. Intraoperative esophagogastroduodenoscopy (IEGD) can be used to assess the orientation and position of a properly constructed Nissen. The aim of this study was to determine if IEGD can replace the routine use of a bougie in the creation of a fundoplication. METHODS: One hundred consecutive patients undergoing laparoscopic Nissen fundoplication from 2003 to 2005 were entered into a prospective database. IEGD was used in all patients instead of a bougie. Preoperative and postoperative data, including symptom scores, pH studies, manometry, and upper gastrointestinal studies, were analyzed. RESULTS: All 100 patients completed the study for a mean follow-up period of 18 months. The mean surgical time was 102 minutes. The mean intraoperative endoscopy time was 14 minutes. There were a total of 24 (24%) alterations of the fundoplication performed according to endoscopic wrap creation. The most common alteration was removal of a fundoplication stitch in 15 patients with a tight appearance of wrap. There were no major complications. Two patients required esophageal dilatation for mild to moderate dysphagia. CONCLUSIONS: IEGD is a valuable tool for laparoscopic Nissen fundoplication. IEGD helps to appreciate the true location of the gastroesophageal junction and allows for fundoplication adjustment based on additional visualization. The creation of a symmetric and floppy wrap during Nissen fundoplication can be facilitated greatly by intraoperative endoscopy and may lead to improved clinical outcomes without the risk of esophageal perforation.

Cellular toxicity of various inhalable metal nanoparticles on human alveolar epithelial cells.

Nanoparticles (NPs) have a greater potential to travel through an organism via inhalation than any other larger particles, and could be more toxic due to their larger surface area and specific structural/chemical properties. The aim of this study was to evaluate in vitro biological effects of various inhalable metallic NPs (TiO2, Ag, Al, Zn, Ni). Human alveolar epithelial cells (A549) were exposed to various concentrations of NPs for 24 h. The extent of morphological damage was in the order of m-TiO2 > n-TiO2 > m-silica >> n-Ni approximately = n-Zn approximately = n-Ag approximately = n-Al and was affected in a dose-dependent manner. The extent of apoptotic damage measured with two-color flow cytometry was in the order of n-Zn > n- Ni > m-silica >> n- TiO2 > m- TiO2 > n-Al > n-Ag. The extent of apoptotic damage measured with DNA fragmentation was in the order of n-Zn approximately = m-silica > n- Ni >> m- TiO2 approximately = n- TiO2 approximately = n-Al > n-Ag, indicating no significant difference in the damages by both m-TiO2 and n-TiO2. The extents of apoptotic damages were also affected in a dose-dependent manner. Uptake of no other NPs but n-TiO2 and m-TiO2 into the cells was observed after 24 h exposure. The intracellular generation of ROS was significant with n-Zn but not with the other particles. These results demonstrated that various inhalable metallic NPs (TiO2, Ag, Al, Zn, Ni) could cause cell damages directly or indirectly. More detailed studies on the influence of size, structure, and composition of the NPs are needed to better understand their toxic mechanisms.

48-Hour pH monitoring increases the risk of false positive studies when the capsule is prematurely passed.

Ambulatory wireless 48-h esophageal pH monitoring (Bravo Medtronic, Shoreview, MN, USA) has been shown to be more sensitive in detecting abnormal esophageal acid exposure compared with transnasal 24-h pH probes. However, accurate interpretation of the wireless monitoring data is paramount when contemplating surgical intervention for those with gastroesophageal reflux disease. The aim of this study is to evaluate the incidence of false-positive interpretations of this wireless monitoring data secondary to premature transit of the Bravo capsule into the stomach and subsequently into the duodenum prior to the completion of the 48-h study period. We reviewed 100 consecutive Bravo pH studies at our University Esophageal Motility Center. There were 58 women and 42 men included in our evaluation. Premature transit of the Bravo capsule into the stomach and subsequently into the small bowel was defined by a prolonged gastric pH phase with either evidence of alkalinization and no further reflux episodes or loss of communication with the Bravo capsule prior to the end of the 48-h data collection period. Of the 100 patients reviewed, 11% manifested evidence of early passage of the Bravo capsule resulting in a misinterpretation of the data as abnormal acid exposure. The mean time of inaccurate data after transit of the Bravo capsule was 18 h and 42 min. The mean length of time that the capsule was retained in the stomach prior to duodenal passage was 4 h. If the aforementioned data were included in the final interpretation of the study, it yielded a mean DeMeester score of 44.25 with a mean total time of pH <4 of 14.7% per case. Exclusion of the prolonged gastric phase from the final interpretation of each case resulted in a statistically significant reduction in the mean total time the pH <4 (4.33 vs. 14.7%, p < 0.05) and the mean DeMeester score (12.81 vs. 44.25 p < 0.05). The mean time from the initiation of esophageal pH data to the passage of the Bravo capsule into the stomach was 15 h and 22 min. The observation mandates meticulous inspection of the pH tracing by the interpreting physician throughout the entirety of a 48-h study to identify premature transit of the capsule. Tracings that show prolonged acid exposure or loss of communication with the Bravo capsule should be screened for the capsule's possible early dislodgement and premature advancement into the stomach.

Cloning and characterization of antioxidant enzyme methionine sulfoxide-S-reductase from Caenorhabditis elegans.

Methionine (Met) residues in proteins are susceptible to oxidation. The resulting methionine sulfoxide can be reduced back to methionine by methionine sulfoxide-S-reductase (MsrA). The MsrA gene, isolated from Caenorhabditis elegans, was cloned and expressed in Escherichia coli. The resultant enzyme was able to revert both free Met and Met in proteins in the presence of either NADPH or dithiothreitol (DTT). However, approximately seven times higher enzyme activity was observed in the presence of DTT than of NADPH. The enzyme had an absolute specificity for the reduction of l-methionine-S-sulfoxide but no specificity for the R isomer. K(m) and k(cat) values for the enzyme were approximately 1.18 mM and 3.64 min(-1), respectively. Other kinetics properties of the enzyme were also evaluated.

Development of a varicella virus vaccine stabilizer containing no animal-derived component.

Various stabilizers containing no animal-derived components were assessed for their efficacy in stabilizing live attenuated varicella virus under various storage temperatures. Stabilizers containing carrageenan, soy protein hydrolysates, or sucrose had a satisfactory performance to maintain infectivity of the cell-free virus when compared to control stabilizer containing animal-derived gelatin/gelatin hydrolysate.