Plasmid curing resulted in improved heterologous gene expression in Leuconostoc citreum EFEL2700.

Leuconostoc citreum EFEL2700 isolated from kimchi was used as a host strain for genetic and metabolic engineering in our previous studies, but the cells of EFEL2700 contained a cryptic plasmid (P-cells). Thus, we created plasmid-free cells (F-cells) using the CRISPR/Cas9 system. In this study, we compared the microbial characteristics of P- and F-cells in terms of growth rate, biochemical properties, transformation efficiency, plasmid copy number and protein expression level. When the growth rate was measured in MRS medium at 30°C, no significant difference (P > 0·01) was observed. Biochemical properties, tested using an API 50CHL kit, showed no differences. Transformation efficiency of F-cells, measured using pCB4270, was higher (1·3 × 104 CFU per µg DNA) than that of P-cells (5·0 × 103 CFU per µg DNA). Copy number after transformation of pCBBgl was 4-fold higher for F-cells than for P-cells. When ß-glucosidase activity was assayed in the above experiment, F-cells showed 3·4-fold higher values than P-cells. In conclusion, this study demonstrates that plasmid curing in L. citreum EFEL2700 improves its characteristics as a gene expression host. SIGNIFICANCE AND IMPACT OF THE STUDY: Leuconostoc citreum EFEL2700 (P-cell) isolated from kimchi is a useful food-grade host for expressing heterologous genes. The presence of a cryptic plasmid is thought to limit efficient gene expression. In this study, we compared the microbial and genetic changes after plasmid curing in this strain. The plasmid-free strain showed improved levels of transformation efficiency, copy number and heterologous gene expression without alterations in phenotypes such as the growth rates and biochemical properties. The resulting strain of L. citreum EFEL2701 (F-cell) can be used as an efficient host for genetic engineering.

Efficacy and safety of non-invasive body tightening with high-intensity focused ultrasound (HIFU).

BACKGROUND: Noninvasive skin-tightening devices have become increasingly popular in response to increasing demand for improvements in skin laxity and tightening with minimal risk and recovery time. OBJECTIVE: We evaluated the efficacy and safety of HIFU for skin tightening in the face and body. METHODS: A total of 32 Korean subjects enrolled in this prospective clinical trial. The subjects were treated with HIFU to both cheeks, lower abdomen, and thigh. Skin elasticity was measured before and after treatment using a Cutometer (CT575, Courage and Khazaka® , Cologne, Germany). Three blinded, experienced dermatologists evaluated paired pre- and post-treatment (week 4 and 12) photographs according to the Global Aesthetic Improvement Scale (GAIS). Participants also completed self-assessments using GAIS. Subjects rated their pain on a numeric rating scale (NRS) immediately, 7 days, 4 weeks, and 12 weeks after treatment. RESULTS: Skin elasticity measured via a Cutometer was significantly improved 12 weeks after treatment at all treated sites (P<.05). Both IGAIS and SGAIS showed significant improvements 12 weeks after treatment. Immediately after treatment the mean NRS score was 3.00±1.586, but no pain was reported at 4 and 12 weeks post-treatment. No serious adverse effects were observed during the follow-up period. CONCLUSION: HIFU safely and effectively improves skin elasticity and clinical contouring of the face and body.

Improved methods for evaluating pre-clinical and histological effects of subcutaneous fat reduction using high-intensity focused ultrasound in a porcine model.

BACKGROUND: Non-invasive body sculpting procedures are becoming increasingly popular. High-intensity focused ultrasound (HIFU) treatment is a non-surgical fat reduction procedure that permanently destroys unwanted abdominal fat. Despite its increasing popularity, evaluation methods for the procedure have not yet been fully developed. AIMS: The objective of this study was to develop evaluation methods for HIFU for non-surgical, permanent fat reduction in the anterior abdomen using a porcine model. METHODS: The abdomens of female pigs (Sus scrofa, n = 7) were treated with a HIFU device (SCIZER™ , Classys Inc, Seoul, Korea). We examined treatment effects using photography, ultrasound, gross and microscopic pathology, and serum lipid and liver function level analysis, carbon tracer test, and histological examination in order to determine the mechanism of action, efficacy, and safety of the procedure. RESULTS: HIFU treatment effectively reduced abdominal fat in a porcine model; it accurately treated the target subcutaneous fat layer and the subcutaneous fat was reduced effectively via ultrasonic measurement after HIFU treatment. On histological staining (H&E, toluidine blue, oil red O and immunohistochemistry), we found that subcutaneous fat reduction occurred effectively via accurate treatment of the targeted subcutaneous fat layer. On hematological assay, there were changes within normal range, and values remained stable after 48 h. Via carbon tracer test, the migration of activated macrophages was identified within the axillary lymph node (LN). PPAR-delta, a protein defined by immunohistochemistry staining, was overexpressed in the early stage on days 1 and 7, but a gradual decreasing pattern was confirmed. CONCLUSION: We successfully used a HIFU device for body contouring and fat reduction in a pre-clinical study. These results provide that the essential clues toward the effective evaluation, guiding selection of the appropriate diagnostic investigations.

An Asian perspective on improving outcomes for nasal bone fractures by establishing specific treatment options.

OBJECTIVE: To report treatment outcomes of patients with different types of nasal bone fracture, following a tailored treatment protocol. DESIGN: The patterns and the severity of the fractures were determined by a preoperative facial photo and nasal bone computed tomography (CT) retrospectively. SETTING: A tertiary referral centre. PARTICIPANTS: We evaluated 129 patients who underwent surgery between March 2002 and January 2014. Patients were subjected to five different treatment methods depending on the severity of injury. MAIN OUTCOME MEASURES: Medical records were reviewed to assess rates of complications and revision surgery. Cosmetic and functional (the degree of nasal obstruction) outcomes were measured using the Likert scale from 1 (very dissatisfied, severe obstruction) to 5 (very satisfied, no obstruction). RESULTS: The mean elapsed time from injury to surgery was 14.9 days. The overall treatment failure rate, defined as a deformity that required revision, was 6.2%. The patient satisfaction scores for aesthetic and functional outcomes were 3.9 and 4.6 respectively. CONCLUSION: The prudent selection of patients with indication for closed reduction, and further specification of variable treatment options for the various degrees of nasal bone fracture, may be helpful in achieving an improved treatment outcome. LEVEL OF EVIDENCE: level IV.

Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model.

Globally, cardiac arrest (CA) is a leading cause of death and disability. Asphyxial CA (ACA)-induced kidney damage is a crucial factor in reducing the survival rate. The purpose of this study was to investigate the role of antioxidant enzymes in histopathological renal damage in an ACA rat model at different time points. A total of 88 rats were divided into five groups and exposed to ACA except for the sham group. To evaluate glomerular function and oxidative stress, serum levels of blood urea nitrogen (BUN) and creatinine (Crtn) and malondialdehyde (MDA) levels in renal tissues were measured. To determine histopathological damage, hematoxylin and eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining were performed. Expression levels of antioxidant enzymes including superoxide dismutase-1 (SOD-1), superoxide dismutase-2 (SOD-2), catalase (CAT), and glutathione peroxidase (GPx) were measured by immunohistochemistry (IHC). Survival rate of the experimental rats was reduced to 80% at 6 h, 55% at 12 h, 42.9% at 1 day, and 33% at 2 days after return of spontaneous circulation. Levels of BUN, Crtn, and MDA started to increase significantly in the early period of CA induction. Renal histopathological damage increased markedly from 6 h until two days post-CA. Additionally, expression levels of antioxidant enzymes were significantly decreased at 6 h, 12 h, 1 day, and 2 days after CA. CA-induced oxidative stress and decreased levels of antioxidant enzymes (SOD-1, SOD-2, CAT, GPx) from 6 h to two days could be possible mediators of severe renal tissue damage and increased mortality rate.

Visualization of superluminal pulses inside a white light cavity using plane wave spatio temporal transfer functions.

In a white light cavity (WLC), the group velocity is superluminal over a finite bandwidth. For a WLC-based data buffering system we recently proposed, it is important to visualize the behavior of pulses inside such a cavity. The conventional plane wave transfer functions, valid only over space that is translationally invariant, cannot be used for the space inside WLC or any cavity, which is translationally variant. Here, we develop the plane wave spatio temporal transfer function (PWSTTF) method to solve this problem, and produce visual representations of a Gaussian input pulse incident on a WLC, for all times and positions.

Distortion free pulse delay system using a pair of tunable white light cavities.

Recently, a tunable bandwidth white light cavity (WLC) was demonstrated by using an anomalously dispersive intra-cavity medium to adjust a cavity linewidth without reducing the cavity buildup factor [G.S. Pati et al., Phys. Rev. Lett. 99, 133601 (2007)]. In this paper, we show theoretically how such a WLC can be used to realize a distortion-free delay system for a data pulse. The system consists of two WLCs placed in series. Once the pulse has passed through them, the fast-light media in both WLCs are deactivated, so that each of these now acts as a very high reflectivity mirror. The data pulse bounces around between these mirrors, undergoing negligible attenuation per pass. The trapped pulse can be released by activating the fast-light medium in either WLC. Numerical simulations show that such a system can far exceed the delay-bandwidth constraint encountered in a typical data buffer employing slow light. We also show that the pulse remains virtually undistorted during the process.

Pro-inflammatory mediators and signaling proteins in the decidua of pre-eclampsia.

OBJECTIVE: To evaluate the role of CD68+ macrophages and inflammatory/signaling proteins in the decidua of singleton pregnancies with late-onset pre-eclampsia. PATIENTS AND METHODS: This study was designed as a prospective case-control study. Decidual tissue samples were obtained from twenty healthy pregnant women as a control group and twenty pregnant women with late-onset pre-eclampsia showing severe symptoms as the study group. We examined the abundance of CD68+ macrophages in both groups using flow cytometry. Protein and mRNA expression levels of inflammatory/signaling proteins, including inducible nitric oxide synthase, nuclear factor-κB inhibitor α, cyclooxygenase-2, and phosphorylated c-Jun N-terminal kinase, in the decidua of both groups were measured using Western blotting and Reverse Transcription-Polymerase Chain Reaction, respectively. Student's t-tests were performed for statistical analysis. RESULTS: The numbers of CD68+ macrophages were similar in the study and control groups (p=0.47). However, the levels of inducible nitric oxide synthase, nuclear factor-κB, cyclooxygenase-2, and phosphorylated c-Jun N-terminal kinase were significantly increased in the study group. Therefore, pro-inflammatory mediators and signaling proteins in the decidua during pre-eclampsia may be related to the pathogenesis of pre-eclampsia. CONCLUSIONS: Pre-eclampsia-induced alterations in the expression of inflammatory/signaling proteins in the decidua during singleton pregnancies may play a critical role in the pathogenesis of pre-eclampsia.

Does isoflurane enhance rhinovirus replication and virus-induced cytokine secretion in airway epithelial cells?

BACKGROUND: There have been no previous studies regarding the effect of volatile anesthetics on human rhinovirus (RV) infection in airway epithelial cells of patients with an upper respiratory infection (URI). We have therefore evaluated in vitro the effect of isoflurane on RV infection in airway epithelial cells. METHODS: A549 cells and RV-infected A549 cells were treated with isoflurane for 2 or 4 h. Surface expression of intercellular adhesion molecule-1 (ICAM-1) was assessed by flow cytometry, and effects on the secretion of interleukin (IL)-6 and IL-8 were measured by ELISA. The effect on RV replication in the cells was determined by viral titer. RESULTS: Isoflurane treatment for 2 or 4 h had no significant effect on ICAM-1 expression and secretion of IL-6 and IL-8 in control cells. Isoflurane also had no significant additional effect on RV-induced ICAM-1 expression and secretion of IL-6 and IL-8. Viral titers were not significantly influenced by isoflurane. CONCLUSION: Isoflurane treatment showed no additional effects with RV on ICAM-1 expression, secretion of IL-6 and IL-8, and viral titer in A549 cells. These results suggest that isoflurane itself may not increase further RV infections, inflammations, and viral replication in patients with a viral URI.

Removal of a metallic foreign body embedded in the external nose via open rhinoplasty approach.

Most metallic foreign bodies are inert, but they can cause chronic inflammatory reactions and be a source of infection. Identification and removal of foreign bodies from wounds is often necessary. The present report describes two cases of a foreign body embedded in the external nose. Each case was successfully treated by an open rhinoplasty approach. This approach is an effective and safe method for removal of foreign bodies in the external nose. It provides a good surgical field and a better cosmetic outcome than a conventional incision.

Endoscopic transnasal reduction of an anterior table frontal sinus fracture: technical note.

Repair of an isolated anterior table frontal sinus fracture traditionally requires a coronal incision, which results in a large scar, alopecia and paresthesias, because of these problems, endoscopic approaches have been attempted. These approaches still involve small incisions at the forehead or behind the hairline so might be better described as endoscopy-assisted surgery. This report describes the reduction of an isolated anterior table frontal sinus fracture in a 14-year-old boy using an endoscopic procedure that involves a transnasal approach with no external incisions. Endoscopic instruments were used under 25 and 70 degrees endoscope guidance. A custom-made latex glove balloon was inserted into the frontal sinus, and then expanded and maintained for 20 days to support the reduced bony fragments. Postoperatively, the reduction was successful. The cosmetic deformity was repaired and there was no postoperative morbidity. This case indicates that endoscopic reduction may be a valuable treatment option for certain types of frontal sinus fractures.

Prevalence of and factors related to latent tuberculous infection among all employees in a referral hospital.

SETTING: Referral hospital, South Korea. OBJECTIVE: To investigate the prevalence of and factors related to latent tuberculous infection (LTBI) among all hospital employees. DESIGN: A cross-sectional study of 479 health care workers (HCWs) undergoing LTBI screening. RESULTS: Overall prevalence of LTBI was 15.7%, 43.1% of whom initiated and completed LTBI treatment. Compared with HCWs without LTBI, those with LTBI were more likely to be older (P < 0.001), male (P = 0.003), work in low-risk departments (P = 0.013) and have more years of employment (P < 0.001). LTBI prevalence was highest in physicians (27.8%), followed by HCWs without patient contact (23.4%), nurses (8.3%) and other HCWs in contact with patients (6.9%). In multivariate analysis, compared with HCWs aged <20 years, those aged 40 years were 4.08 times more likely to have LTBI (P = 0.007). In addition, compared with HCWs working for <1 year, those working for 1-5 years or for 5 years were respectively 7.55 (P = 0.014) and 13.69 (P = 0.001) times more likely to have LTBI. CONCLUSIONS: Our results suggest that modified LTBI screening strategies, including HCWs with no patient contact and encouraging LTBI treatment participation, might be helpful in improving LTBI control in HCWs.

Anti-asthmatic effects of volatile organic compounds from Chamaecyparis obtusa, Pinus densiflora, Pinus koraiensis, or Larix kaempferi wood panels.

Asthma is a common chronic inflammatory disease in which lung airways narrow and produce extra mucus. Numerous medications, such as steroids, are used to prevent or control asthmatic symptoms, but side effects are associated with those medications. There are reports of anti-inflammatory, antibacterial, and antiparasitic effects of terpene, a volatile organic compound (VOC) in conifers. VOCs easily enter a gaseous form, and wood products are good sources of VOCs. However, only a few studies have been conducted on the effect on asthma of VOCs emitted by wood. In this study, we examined the effects of VOCs diffused from wood panels on ovoalbumin (OVA)-induced asthma in a mouse model. The mice were intraperitoneally sensitized with 10 µg of OVA with aluminum hydroxide on days 0, 7, and 14. From day 21 to day 26, the mice were challenged with 2% OVA intranasally for 30 min. For VOC treatment, asthma model mice were placed in polyacrylamide chambers containing wood panels of Chamaecyparis obtusa, Pinus densiflora, Pinus koraiensis, or Larix kaempferi. On day 27, serum, lung tissue, and bronchoalveolar lavage fluids were prepared for H&E staining, qRT-PCR, ELISA, and Diff-Quik staining, as appropriate. OVA treatment induced hypertrophy of the bronchiolar wall. The budesonide group and all four of the wood panel-exposed groups showed less thickening of the bronchiolar wall and downregulated transcriptional expressions of cytokines such as interleukin-4 (IL-4) and interleukin-13 (IL-13). The serum tumor necrosis factor-α (TNF-α) mRNA expression level was significantly decreased only in the C. obtusa group, but the serum IL-4 levels were decreased in all wood panel treatment groups. Diff-Quik staining of bronchoalveolar lavage fluids revealed a decrease in the number of granulocytes in all wood panel treatment groups. The results suggest that VOCs from C. obtusa, P. densiflora, P. koraiensis and L. kaempferi produce antiasthmatic effects by regulating the production of IL-4, IL-9, IL-13, TNF-α.

Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model

Globally, cardiac arrest (CA) is a leading cause of death and disability. Asphyxial CA (ACA)-induced kidney damage is a crucial factor in reducing the survival rate. The purpose of this study was to investigate the role of antioxidant enzymes in histopathological renal damage in an ACA rat model at different time points. A total of 88 rats were divided into five groups and exposed to ACA except for the sham group. To evaluate glomerular function and oxidative stress, serum levels of blood urea nitrogen (BUN) and creatinine (Crtn) and malondialdehyde (MDA) levels in renal tissues were measured. To determine histopathological damage, hematoxylin and eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining were performed. Expression levels of antioxidant enzymes including superoxide dismutase-1 (SOD-1), superoxide dismutase-2 (SOD-2), catalase (CAT), and glutathione peroxidase (GPx) were measured by immunohistochemistry (IHC). Survival rate of the experimental rats was reduced to 80% at 6 h, 55% at 12 h, 42.9% at 1 day, and 33% at 2 days after return of spontaneous circulation. Levels of BUN, Crtn, and MDA started to increase significantly in the early period of CA induction. Renal histopathological damage increased markedly from 6 h until two days post-CA. Additionally, expression levels of antioxidant enzymes were significantly decreased at 6 h, 12 h, 1 day, and 2 days after CA. CA-induced oxidative stress and decreased levels of antioxidant enzymes (SOD-1, SOD-2, CAT, GPx) from 6 h to two days could be possible mediators of severe renal tissue damage and increased mortality rate.

Percutaneous direct puncture glue embolization of high-flow craniofacial arteriovenous lesions: a new circular ring compression device with a beveled edge.

SUMMARY: The purpose of this study was to evaluate the efficacy of a newly designed circular ring compression device that allows safe and effective glue injection during preoperative embolization of high-flow superficial craniofacial arteriovenous malformations (AVMs). The device was used in 4 cases of craniofacial AVM with multiple feeding arteries and draining veins. It provided a safe glue injection route as well as effective compression of radiating multiple venous drainage routes.

Donor site morbidities resulting from conchal cartilage harvesting in rhinoplasty.

OBJECTIVE: Conchal cartilage is frequently used in rhinoplasty, but donor site morbidity data are seldom reported. This study aimed to investigate the complications of conchal cartilage harvesting in rhinoplasty. METHODS: A retrospective chart review of 372 patients who underwent conchal cartilage harvesting for rhinoplasty was conducted. Data regarding patient demographics, types of nasal deformities, graft usage and complications were analysed. RESULTS: A total of 372 patients who underwent conchal cartilage harvesting for rhinoplasty were enrolled. The harvested conchal cartilage tissues were used in a variety of applications: tip graft, dorsal graft, septal reinforcement and correction of nostril asymmetry. Nine cases (2.4 per cent) with donor site morbidities were identified, including four cases (1.1 per cent) with keloids and five cases (1.3 per cent) with haematomas. CONCLUSION: Conchal cartilage harvesting is a safe and useful technique for rhinoplasty, with a low complication rate. However, patients should be informed about the possibility of donor site morbidities such as keloids and haematomas.

p53 mediated sensitization of squamous cell carcinoma of the head and neck to radiotherapy.

Radiation resistant squamous cell carcinoma of the head and neck cell line JSQ-3 carries a mutant form of tumor suppressor gene p53. Treatment of these cells with an adenoviral vector containing wild-type p53 (Av1p53) was able to inhibit their growth in vitro and in vivo while having no effect on normal cells. More significantly, introduction of wtp53 also reduced the radiation-resistance level of this cell line in vitro, in a viral dose-dependent manner. Furthermore, this radiosensitization also carried over to the in vivo situation where the response of JSQ-3 cell-induced mouse xenografts to radiotherapy was markedly enhanced after treatment with Av1p53. Complete, long-term regression of the tumors for up to 162 days was observed when a single dose of Av1p53 was administered in combination with ionizing radiation, demonstrating the effectiveness of this combination of gene therapy and conventional radiotherapy. This sensitization of tumors to radiation therapy by replacement of wtp53 could significantly decrease the rate of recurrence after radiation treatment. Since radiation is one of the most prevalent forms of adjunctive therapy for a variety of cancers, these results have great relevance in moving toward an improved cancer therapy.

Fission yeast dihydrolipoamide dehydrogenase gene is involved in G1/S cell cycle progression.

Using functional complementation with a Schizosaccharomyces pombe genomic library, we have isolated a clone complementing a G1/S phase progression defective mutant. The newly isolated temperature-sensitive mutant, cyj150, showed elongated morphology at a restrictive temperature of 36 degrees C and DNA content analysis of the mutant indicated a defect in cell cycle progression at the G1/S phase. Sequence analysis of the genomic and cDNA clones complementing this elongated phenotype at 36 degrees C show that it encodes a protein that has 50% amino acid identity with dihydrolipoamide dehydrogenase from Saccharomyces cerevisiae and garden pea. Alignment of the deduced amino acid sequence of S. pombe dihydrolipoamide dehydrogenase (dld1+) with glutathione reductase and mercuric reductase revealed extensive homologies throughout the primary sequence and protein structure, and contained amino acid sequences of the active site region conserved from prokaryote to higher eukaryote. Gene disruption and tetrad analysis showed that dld1+ is an essential gene for cell viability. Northern analysis indicates that transcriptional expression of this gene is not fluctuated according to the cell cycle. However, it is certain that malfunction of this Dld1 protein blocks the progression of cell cycle from G1 to S phase. The sequence of the dld1+ gene is available in EMBL/GenBank under Accession Number L40360.

Alternative splicing in the Caenorhabditis elegans DNA topoisomerase I gene.

5'-end cDNA fragments of the Caenorhabditis elegans DNA topoisomerase I gene were obtained by rapid amplification of the cDNA ends from C. elegans mRNAs. The presence of a SL1 sequence at the 5'-terminus of the cDNA sequence suggested trans-splicing of the pre-mRNA. By comparing the complete cDNA sequence with the genomic lambda DNA clones, the gene structure composed of five exons was established. Alternative splicing deleting the second exon was observed in the cDNA fragments obtained by a gene-specific reverse transcription followed by polymerase chain reactions. The shorter mRNA missing the second exon was expressed at all the developmental stages, while the full-length mRNA was present only in embryos.