Cataract surgery with new monofocal intraocular lens enhanced for intermediate vision and standard monofocal intraocular lens for retinal disorder.

AIM: To investigate the difference in the initial surgical results between a new monofocal intraocular lens (IOL) with enhanced intermediate vision and the standard monofocal IOL in patients with retinal disease. METHODS: We retrospectively reviewed the medical records of patients with retinal disease who underwent cataract surgery due to accompanying cataracts. Types of retinal diseases were investigated and best-corrected distant visual acuity, distant uncorrected visual acuity (UCVA), intermediate UCVA, near UCVA, and spherical equivalent were recorded at each visit. The surgical results were investigated at 1 day, 1 week, and 1 month after surgery. RESULTS: Seventeen eyes treated with a new monofocal IOL enhanced for intermediate vision (ICB00 group) and 18 eyes treated with the standard monofocal IOL (AAB00 group) were included in this study. There were no significant differences in the baseline characteristics, including the type of underlying retinal disease, between the groups. There were no significant differences between the groups in terms of distant, intermediate, or near UCVA at day 1 and week 1 after surgery. However, at 1 month after surgery, the ICB00 group showed a significantly better intermediate vision improvement than the AAB00 group (p = 0.001). CONCLUSION: Even in patients with cataract accompanied by retinal disease, the use of the ICB00 IOL showed significant improvement in intermediate vision compared to the use of the AAB00 (standard monofocal) IOL. The ICB00 IOL might be a good option for patients with cataract and retinal disease in the era of increased intermediate vision needs in daily life.

Anti-inflammatory and anti-apoptotic effects of N-acetylcysteine in diabetic rat corneal epithelium.

AIM: To characterize the anti-inflammatory and anti-apoptotic effects of N-acetylcysteine (NAC) in streptozotocin (STZ)-induced diabetic rat corneal epithelium and human corneal epithelial cells (HCECs) exposed to a high-glucose environment. METHODS: HCECs were incubated in 0, 5, 50 mmol/L glucose medium, or 50 mmol/L glucose medium with NAC for 24h. Diabetes was induced in rats by intraperitoneal injection of 65 mg/kg STZ and some of these rats were topically administered NAC to corneas with 3 mice per group. We characterized receptor for advanced glycation end-products (RAGE) expression using immunofluorescence, and interleukin (IL)-1ß and cleaved caspase-3 (CCAP-3) expression using immunohistochemistry. Circulating tumor necrosis factor (TNF)-α concentration was measured by ELISA and cleaved poly-ADP ribose polymerase (PARP) concentration was quantified by Western blotting. Apoptotic cells were detected using TUNEL assay and annexin V and propidium iodide staining. RESULTS: Diabetic rats had higher expression of RAGE (2.46±0.13 fold), IL-1ß, and CCAP-3 in apoptotic cells of their corneas than control rats. The expression of RAGE (1.83±0.11 fold), IL-1ß, and CCAP-3, and the number of apoptotic cells, were reduced by topical NAC treatment. HCECs incubated in 50 mmol/L glucose medium showed high concentrations of TNF-α (310±2.00 pg/mL) and cleaved PARP (7.43±0.56 fold), and more extensive apoptosis than cells in 50 mmol/L glucose medium. However, the addition of NAC reduced the concentrations of TNF-α (153.67±2.31 pg/mL) and cleaved PARP (5.55±0.31 fold) and the number of apoptotic cells. CONCLUSION: NAC inhibits inflammation and apoptosis in the corneas of diabetic rats and HCECs maintained in a high-glucose environment.

Topical nerve growth factor attenuates streptozotocin-induced diabetic cataracts via polyol pathway inhibition and Na+/K+-ATPase upregulation.

The purpose of this study was to investigate whether and how topical nerve growth factor (NGF) attenuates streptozotocin (STZ)-induced diabetic cataracts in vivo. Rats were randomly divided into three groups, including the normal control rat group, STZ-induced diabetic cataract rat group (DM group), and STZ-induced diabetic cataract rat group treated with 200 µg/mL recombinant rat ß-NGF (DM + NGF group). Cataract formation was evaluated by portable slit lamp biomicroscopy following pupil dilation at 8 weeks. The expression levels of NGF, aldose reductase (AR), and Na+/K+-ATPase in the lens epithelial cells (LECs) of the three groups were measured in the presence or absence of topical NGF. TUNEL-positive LECs were quantified to determine if hyperglycemia caused LEC apoptosis. At 8 weeks, the mean cataract score in the control group was significantly lower than that in DM and DM + NGF groups, and the score in the DM + NGF group was significantly lower than that in the DM group. At the equatorial zone and anterior central zone of lens, NGF and Na+/K+-ATPase expression levels were significantly decreased in the DM group; however, they were partially restored in the DM + NGF group. At the equatorial zone and anterior central zone of lens, AR expression and TUNEL-positive apoptotic LECs were significantly increased in the DM group compared with the control group, however, they were significantly decreased in the DM + NGF group. In conclusion, topical NGF could delay the progression of diabetic cataracts by attenuating polyol pathway activation and increasing Na+/K+-ATPase protein levels.

Diffractive multifocal intraocular lens implantation in patients with monofocal intraocular lens in the contralateral eye.

AIM: To evaluate clinical outcomes of unilateral implantation of a diffractive multifocal intraocular lens (IOL) in patients with contralateral monofocal IOL. METHODS: Twenty-two patients who already had implantation of a monofocal IOL in unilateral eye underwent implantation of a diffractive multifocal IOL in contralateral eye were enrolled. After 1, 6, and 12mo, uncorrected and distant corrected distant visual acuity (UCDVA and DCDVA), uncorrected and distant corrected intermediate-visual acuity (UCIVA and DCIVA), uncorrected and distant corrected near visual acuity (UCNVA and DCNVA), and contrast sensitivity were obtained. Halo/glare symptoms, spectacle dependence, and patient satisfaction were also evaluated. RESULTS: The mean age was 67.86±7.25y and the average interval between two IOL implantations was 645.82±878.44d. At 1mo, binocular UCDVA was lower than 0.20 logMAR in 76% of patients (mean 0.12±0.13 logMAR), which increased to 90% by 6 and 12mo. The binocular UCDVA was significantly better than the monocular results (P<0.05) at 1, 6, and 12mo. Additionally, UCNVA was lower than 0.40 logMAR in 82% of patients, increasing to 90% by 6 and 12mo. Mean UCNVA in the multifocal IOL implanted eye was statistically significantly better than that in the monofocal IOL implanted eye (P<0.05) at 1, 6, and 12mo. About 5% of patients at 1 and 6mo, reported "severe glare or halo". Patient satisfaction rates were 95% and 91% at 6 and 12mo, respectively. CONCLUSION: Unilateral implantation of multifocal IOL in patients with a contralateral, monofocal IOL implantation results in high patient satisfaction rate, with low severe glare or halo rate during follow-up. It can represent a good option for patients who have previously had a monofocal IOL implantation regardless of two year interval duration between two IOL implantations.

Diquafosol Sodium Inhibits Apoptosis and Inflammation of Corneal Epithelial Cells Via Activation of Erk1/2 and RSK: In Vitro and In Vivo Dry Eye Model.

Purpose: To evaluate the effect of diquafosol on corneal epithelium in a dry eye model using Transwell culture and a scopolamine-induced dry eye rat model. Methods: Desiccation stress induced in an in vitro dry eye model using human corneal epithelial cells was used, and the cells were incubated with or without diquafosol media diluted at 1:100. Reactive oxygen species (ROS) generation was measured using 2',7'-dichlorofluorescein diacetate (DCFH-DA). Apoptosis was analyzed, and levels of phosphorylated Erk1/2, phosphorylated p90RSK, phosphorylated Akt, IκB-α, and NF-κB-p65 were determined. Levels of IL-1ß, TNF-α, IL-6, IL-8, and GM-CSF were quantified. To investigate the in vivo effects of diquafosol, we induced dry eye in Wistar rats using scopolamine hydrobromide. The rats were divided into three groups: control, dry eye, and dry eye diquafosol; topical DIQUAS was applied four times daily for 28 days. We used immunohistochemistry to detect the levels of phosphorylated Erk1/2, phosphorylated p90RSK, and IL-1ß, and used the TUNEL assay in corneal tissue. Results: The distribution of highly fluorescent dichlorofluorescein and the proportion of annexin V- and PI-positive cells decreased in the diquafosol medium. Diquafosol increased the levels of phospho-Erk1/2, phospho-90RSK, phospho-Akt, and IκB-α, whereas it significantly decreased the levels of NF-κB-p65, IL-1ß, and TNF-α. In vivo, apoptosis was enhanced in dry eye group. This response was markedly reduced and the level of phosphorylated p90RSK and phosphorylated ERK1/2 were upregulated and IL-1ß was downregulated by DIQUAS. Conclusions: Diquafosol treatment reduced intracellular ROS levels, apoptosis, and inflammation, all of which were increased in the dry eye model through desiccation.

Effects of propofol and isoflurane on excitatory amino acid carrier 1 mRNA and glutathione protein levels in rat hippocampus.

OBJECTIVE: We compared the effects of two anesthetics, isoflurane and propofol, on the nuclear or cytosolic localization of nuclear factor erythroid 2-related factor 2 (Nrf2), mRNA expression levels of excitatory amino acid carrier 1 (EAAC1), and glutathione (GSH) protein levels in the rat hippocampus. METHODS: Fifty-two adult male Sprague-Dawley rats were randomly divided into three groups: a control group, a group that received propofol for 240 minutes (P240), and a group that received isoflurane for 240 minutes (I240). We compared GSH protein and EAAC1 mRNA expression levels in the rat hippocampus and evaluated Nrf2 content in cytosolic and nuclear fractions in the three groups. RESULTS: GSH protein and EAAC1 mRNA expression levels were significantly higher in the I240 and P240 groups compared with the control group. The I240 and P240 groups showed lower Nrf2 protein levels in the cytosolic fractions, but higher levels in the nuclear fractions compared with the control group. CONCLUSION: Treatment with isoflurane or propofol may enhance GSH production by facilitating translocation of Nrf2 into the nucleus and increasing EAAC1mRNA expression in the rat hippocampus. Isoflurane and propofol show similar profiles in EAAC1 expression-associated GSH production.

Comparison of immediate small-incision lenticule extraction after suction loss with uneventful small-incision lenticule extraction.

PURPOSE: To evaluate and compare the predictability and safety of immediate small-incision lenticule extraction, also known as SMILE, after suction loss during the procedure with those of uneventful small-incision lenticule extraction. SETTING: Nunemiso Eye Center, Seoul, South Korea. DESIGN: Retrospective case series. METHODS: Patients who had immediate small-incision lenticule extraction using reapplied suction (study group) or uneventful small-incision lenticule extraction (control group) for myopia or myopic astigmatism were included. All patients were followed for at least 12 months. Demographic as well as visual and refractive outcomes were compared between the 2 groups. RESULTS: Twenty-three eyes were enrolled in the study group and 48 eyes in the control group. The overall cumulative incidence of suction loss was 0.2% (28 of 11 762 eyes). Twenty-two eyes (96%) in the study group had unchanged or improved decimal Snellen corrected distance visual acuity (CDVA). At 12 months, the mean uncorrected distance visual acuity, CDVA, and manifest refraction defocus equivalent in the study group and control group were 0.99 ± 0.17 (SD), 1.11 ± 0.14, 0.75 ± 0.50 diopters (D), and 1.25 ± 0.22 D, 1.26 ± 0.21 D, 0.35 ± 0.26 D, respectively (P = .000, P = .007, and P = .000, respectively). The correlation between the achieved and the attempted spherical equivalent refraction was slightly better in the control group (R2 = 0.972) than in the study group (R2 = 0.933). CONCLUSIONS: Immediate small-incision lenticule extraction using reapplied suction resulted in safe and clinically predictable long-term outcomes. However, predictability was lower for immediate than for uneventful small-incision lenticule extraction.

Nerve Growth Factor Attenuates Apoptosis and Inflammation in the Diabetic Cornea.

Purpose: To examine the effects of nerve growth factor (NGF) on apoptosis and inflammation in the diabetic cornea. Methods: To investigate the effects of NGF on glucose-induced apoptosis, we stained human corneal epithelial cells (HCECs) for annexin-V and propidium iodide (PI), and measured expression of cleaved caspase-3 and the Bcl-2-associated X protein (BAX). Moreover, to examine the effects of NGF on inflammation, we quantified the expression of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) using multiplex cytokine analysis, and analyzed nuclear factor-κB (NF-κB) activation and NF-κ-B inhibitor α (IκBα) degradation using Western blot analysis. To investigate the effects in vivo, we induced diabetes in male Sprague-Dawley rats using streptozotocin. The rats were divided into three groups: control, diabetic control, and diabetic NGF; topical NGF was applied three times daily for 3 weeks. We used the TUNEL assay to detect apoptosis in corneal tissue, and immunohistochemistry to identify cleaved caspase-3 and IL-1ß. Results: In HCECs, high glucose concentration (≥25 mM) led to reactive oxygen species (ROS) generation, apoptosis, and the release of inflammatory cytokines. Nerve growth factor markedly reduced ROS activation, annexin-PI-positive cells, and levels of cleaved caspase-3, BAX, IL-1ß, and TNF-α. In the diabetic rat cornea, apoptosis and inflammation were enhanced, as were levels of cleaved caspase-3 and IL-1ß. These responses were markedly reduced by NGF. Conclusions: Apoptosis and inflammation are enhanced in the diabetic cornea; NGF attenuates these responses-both in vivo and in vitro. Therefore, NGF therapy may represent a novel approach for the management of diabetic keratopathy.

Preoperative depressed mood and perioperative heart rate variability in patients with hepatic cancer.

STUDY OBJECTIVE: How perioperative heart rate variability (HRV) indices differ according to the anxiety or depressed mood of patients scheduled to undergo a major surgical procedure for cancer. DESIGN: Prospective observational study. SETTING: Operating room. PATIENTS: Forty-one male patients between 40 and 70 years of age with hepatocellular carcinoma were included in the final analysis. INTERVENTIONS: HRV was measured on the day before surgery (T1), impending anesthesia (T2), and after anesthetic induction (T3). Preoperative anxiety and depressed mood of all patients were evaluated using the State-Trait Anxiety Inventory and Self-Rating Depression Scale (SDS). MEASUREMENTS AND RESULTS: HRV was significantly different among T1, T2, and T3. At T2, high frequency (HF) (normalized units of HF [nuHF]) was decreased and low frequency (LF) (normalized units of LF) and LF/HF were increased compared with those at T1 and T3. In the subgroup analysis between high and low SDS groups, high SDS group showed significantly decreased nuHF (P = .035), increased nuLF (P = .039), and increased LF/HF (P = .020) compared to low SDS group at T1. However, these values at T2 and T3 were not different between 2 groups. In analysis within the groups, low SDS group showed significant differences in nuHF, nuLF, and LF/HF among T1, T2, and T3 (P < .05, respectively), but no changes in these values were observed in high SDS group among the 3 different time points. CONCLUSIONS: HRV decreased significantly immediately before anesthesia and recovered to baseline with anesthetic induction. Preoperative, more depressed patients showed increased sympathetic tone at baseline and blunted response to impending anesthesia on the HRV measurements.

Transesophageal echocardiographic guidance for percutaneous closure of aortic pseudoaneurysm using a type II Amplatzer vascular plug: a case report.

Aortic pseudoaneurysm after cardiac surgery is a rare entity, but it is potentially fatal due to its clinical course along with higher morbidity and mortality rates. Instead of open surgical repair, percutaneous procedures have been introduced as other options for managing an aortic pseudoaneurysm. In this case report, we describe transesophageal echocardiography guidance for successful percutaneous closure of an aortic pseudoaneurysm located in the left ventricular outflow tract by using a type II Amplatzer vascular plug in a patient in whom open surgical repair was not recommended.

Small-incision lenticule extraction in posterior polymorphic corneal dystrophy.

UNLABELLED: We present the case of a 37-year-old man with posterior polymorphic corneal dystrophy who had small-incision lenticule extraction surgery to correct myopic refractive errors. The subjective refraction was -2.62 in the right eye and -2.50 -0.25 × 80 in the left eye. Slitlamp biomicroscopy revealed a band-like lesion in the corneal endothelium of the right eye. The endothelial cell count was 1745 cells/mm(2) in the right eye and 2945 cells/mm(2) in the left eye. Small-incision lenticule extraction was performed to correct the myopic refractive errors in both eyes. Twelve months after surgery, the uncorrected distance visual acuity in both eyes was 20/20 and the posterior polymorphic corneal dystrophy lesion had not progressed. The case suggests that small-incision lenticule extraction surgery can be performed successfully and safely in patients with posterior polymorphic corneal dystrophy. FINANCIAL DISCLOSURE: None of the authors has a financial or proprietary interest in any material or method mentioned.

Corneal coma and trefoil changes associated with incision location in cataract surgery.

PURPOSE: To investigate changes in corneal higher-order aberrations (HOAs) associated with the incision location in cataract surgery. SETTING: Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea. DESIGN: Observational case series. METHODS: Eyes were divided into 3 groups according to the different axes of incision location: superior, temporal, and nasal. Preoperative and 1-month postoperative corneal coma, trefoil, and spherical aberrations were compared. RESULTS: The study evaluated 119 eyes that had uneventful cataract surgery with a 2.2 mm limbal incision. The superior incision group showed significant changes in the vertical coma (amount of change: -0.119 µm ± 0.153 [SD]; P < .001) and vertical trefoil (amount of change: 0.185 ± 0.260 µm, P = .001). The nasal incision group showed a significant change in oblique trefoil (amount of change: 0.176 ± 0.207 µm; P < .001). The temporal incision group did not show significant changes in HOAs. CONCLUSION: The superior incision in 2.2 mm phacoemulsification cataract surgery caused a change in the corneal vertical coma in a negative direction. Corneal trefoil changes were dependent on the incision location. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Accuracy of an automated refractor using a Hartmann-Shack sensor after corneal refractive surgery and cataract surgery.

PURPOSE: To evaluate the accuracy of a new automated refractor (HRK-8000A) using the Hartmann-Shack sensor in eyes with previous corneal refractive surgery or intraocular lens (IOLs) implantation. SETTING: Asan Medical Center, Seoul, South Korea. DESIGN: Retrospective nonrandomized clinical study. METHODS: Refractive errors were evaluated after more than 1 month postoperatively. Subjective refractive error measurements were obtained by the same practitioner. Refractive error measurements were obtained with the Hartmann-Shack sensor refractor and a conventional refractor (KR 8800). RESULTS: There were 121 eyes (121 patients) with previous corneal refractive surgery and 122 eyes (122 patients) with previous cataract surgery. The results of refractive error measured by the Hartmann-Shack sensor refractor were more similar to subjective refraction than those measured by the conventional refractor in both groups. In both groups, the mean spherical equivalent (SE) for the subjective refraction showed better agreement with the automated refraction measured by the Hartmann-Shack sensor refractor (limits of agreement [LoA], -0.66 to +0.65 diopters [D] and -0.47 to +0.41 D, respectively) than the automated refraction measured by the conventional refractor (LoA, -1.66 to +0.58 D and -1.34 to +0.60 D, respectively). In both groups, the Jackson cross-cylinder at axis 0 degrees and 45 degrees from the automated refraction by the Hartmann-Shack sensor refractor and conventional refractor showed similar agreement with the subjective refraction. CONCLUSION: The Hartmann-Shack sensor automated refractor gave reliable and valid objective refraction results in patients who have had cataract or corneal refractive surgery. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

A new multiple noncontinuous puncture (pointage) technique for corneal tattooing.

AIM: To assess the safety and cosmetic efficacy of a new multiple noncontinuous transepithelial puncture technique for tattooing a decompensated cornea. METHODS: It was a non-comparative clinical case series study. The study examines 33 eyes in 33 patients with total corneal opacity due to corneal decompensation, which developed following intraocular surgery. Corneal tattooing was performed using the multiple noncontinuous transepithelial puncture technique (i.e. pointage). The safety of this new surgical strategy was assessed by occurrence of adverse events for the follow-up period. The cosmetic efficacy was determined by the patient's cosmetic satisfaction and independent observer's opinion about patient appearance. RESULTS: Seven women and 26 men were included in the study. The mean age was 46.4±17.5y (range: 7-67). In total, 30 of 33 patients (91%) reported cosmetic satisfaction within the follow-up period. Only 3 patients (9%) required additional tattooing due to cosmetic unsatisfaction. Cosmetic outcomes were analyzed and classified as excellent or good in 13 (39%) and 17 (52%) patients, respectively. No serious adverse events developed, except delayed epithelial healing in 3 cases. CONCLUSION: The cosmetic outcomes of the multiple noncontinuous transepithelial puncture technique for corneal tattooing were good. The safety of this method is higher than conventional procedures. This new procedure also provides improved cost-effectiveness and safety over current corneal tattooing techniques.

The Antioxidant N-Acetylcysteine Inhibits Inflammatory and Apoptotic Processes in Human Conjunctival Epithelial Cells in a High-Glucose Environment.

PURPOSE: We evaluated the effects of N-acetylcysteine (NAC), which is known to inhibit reactive oxygen species (ROS)-dependent apoptosis, on high glucose-induced ROS, apoptosis, inflammation, and delayed-wounding closure in primary cultured human conjunctival epithelial cells (pHCECs), and the regulatory effects of cleaved caspase-3, BAX, nuclear factor κB (NF-κB), IL-6, and TNF-α on these processes. METHODS: High glucose-induced ROS generation was measured using 2',7'-dichlorofluorescein diacetate (DCFH-DA). The effects of NAC on high glucose-induced apoptosis were investigated in pHCECs using Annexin-V and PI staining, and cleaved caspase-3 and BAX expression levels using immunoblotting. To evaluate the inflammatory response, IL-6 and TNF-α expression levels were quantified by multiplex cytokine analysis, and NF-κB activation and IkB-α degradation were assessed by Western blot analysis. The effects of NAC on high glucose-delayed conjunctival epithelial wound healing were assessed by a scratch-induced directional wounding assay. RESULTS: Compared to the untreated control and normal glucose (5 mM), high glucose at 25 mM stimulated ROS generation, apoptosis, and release of inflammatory cytokines, and delayed wound healing in pHCECs. The addition of NAC markedly reduced the high glucose-induced ROS activation, Annexin-PI-positive cells, and levels of cleaved caspase-3, BAX, IL-6, and TNF-α. N-acetylcysteine also prevented high glucose-delayed wound healing. CONCLUSIONS: High glucose levels promote apoptosis by affecting mitochondria-dependent caspase activity through elevation of ROS production, a process that can be reversed by the antioxidant NAC. These findings demonstrate that NAC has a beneficial effect on conjunctival epithelial cell wound healing, antiapoptosis, and anti-inflammation in the conjunctival epithelial cell.

Efficacy and safety of combination treatment for oculodermal melanocytosis: surgical reduction and use of 532-nm Q-switched Nd: YAG laser.

PURPOSE: The aim of this study was to evaluate the efficacy and safety of combination treatment using surgical reduction technique and 532-nm Q-switched Nd:YAG laser treatment for the removal of oculodermal melanocytosis. METHODS: We retrospectively reviewed the medical records of patients who underwent combination treatment using surgical reduction and 532-nm Q-switched Nd:YAG laser for the removal of ocular pigmentation in oculodermal melanocytosis at Asan Medical Center (Seoul, Korea) between January 2009 and January 2013. The safety and cosmetic efficacy of this new surgical strategy were assessed by both patients and observers. RESULTS: Fifty eyes of 47 patients with oculodermal melanocytosis were enrolled. Ocular pigmentation was successfully reduced in all patients, and all patients were satisfied with the cosmetic results at the last visit. Preoperative and postoperative corrected visual acuity, keratometry, and refractive errors did not significantly differ. Postoperative intraocular pressure was significantly reduced (mean change, 2.33 ± 3.54 mm Hg; P = 0.03) at 1 month after surgery. During the follow-up period, no serious adverse events developed other than conjunctival neovascularization and conjunctival inclusion cyst in 5 eyes, respectively. CONCLUSIONS: Combination treatment for oculodermal melanocytosis removal seems to be cosmetically satisfactory and safe. This study suggested that combination treatment using surgical technique and 532-nm Q-switched Nd:YAG laser may be helpful for the removal of oculodermal melanocytosis.

Anti-cell death engineering of CHO cells: co-overexpression of Bcl-2 for apoptosis inhibition, Beclin-1 for autophagy induction.

Genetic engineering approaches to inhibit cell death in Chinese hamster ovary (CHO) cell cultures have been limited primarily to anti-apoptosis engineering. Recently, autophagy has received attention as a new anti-cell death engineering target in addition to apoptosis. In order to achieve a more efficient protection of cells from the stressful culture conditions, the simultaneous targeting of anti-apoptosis and pro-autophagy in CHO cells (DG44) was attempted by co-overexpressing an anti-apoptotic protein, Bcl-2, and a key regulator of autophagy pathway, Beclin-1, respectively. Co-overexpression of Bcl-2 and Beclin-1 exhibited a longer culture period as well as higher viability during serum-free suspension culture, compared with the control (without co-overexpression of Bcl-2 and Beclin-1) and Bcl-2 overexpression only. In addition to the efficient inhibition of apoptosis by Bcl-2 overexpression, Beclin-1 overexpression successfully induced the increase in the autophagic marker protein, LC3-II, and autophagosome formation with the decrease in mTOR activity. Co-immunoprecipitation and qRT-PCR experiments revealed that the enforced expression of Beclin-1 increased Ulk1 expression and level of free-Beclin-1 that did not bind to the Bcl-2 despite the Bcl-2 overexpression. Under other stressful culture conditions such as treatment with sodium butyrate and hyperosmolality, co-overexpression of Bcl-2 and Beclin-1 also protected the cells from cell death more efficiently than Bcl-2 overexpression only, implying the potential of autophagy induction. Taken together, the data obtained here provide the evidence that pro-autophagy engineering together with anti-apoptosis engineering yields a synergistic effect and successfully enhances the anti-cell death engineering of CHO cells.

Evaluation of stress patterns generated by reduction forceps within a photoelastic mandibular model.

INTRODUCTION: Little attention has been paid to the mechanical effects of fracture reduction forceps. AIM: This study aims to evaluate the stress patterns within the fractured mandible generated by reduction forceps. MATERIAL AND METHODS: Thirty-six mandibular models were fabricated using a photoelastic resin. Each of the three sets of mandibular models was osteotomized according to one of three different fracture types. After reducing the cut segments, reduction forceps were placed into different engagement holes to compress the segments. Photoelastic stress analysis was used to visualize the stress patterns within the fractured mandibular models as generated by the reduction forceps. RESULTS: In the case of symphyseal or parasymphyseal fractures, an optimum distribution of stresses over the fracture site was achieved when placing the reduction forceps more than 12mm away from either side of the fracture line, between the midway level of the mandibular height (bisecting the mandible) and 5mm below this level. In the case of body fractures, optimum stress distribution was achieved when the reduction forceps were placed more than 16mm from the fracture line at the midway level. CONCLUSION: Correct use of the reduction forceps helps to provide a precise three-dimensional reduction for mandibular fractures.