Jae-Young Cho

Lysophosphatidic acid stimulates CREB through mitogen- and stress-activated protein kinase-1.

Lysophosphatidic acid (LPA) is a growth factor-like phospholipid that elicits a variety of cellular responses in numerous cell types, including neurons, immune cells, and fibroblasts. In this report, we investigated the possibility that LPA activates the transcription factor cAMP response element-binding protein, CREB, in Rat-2 fibroblast cells. CREB is activated in many cells downstream of signaling events, such as growth factor and neurotrophin stimulation. We found that LPA rapidly stimulated phosphorylation of CREB at Ser133 in a time- and dose-dependent manner, as revealed by immunoblot analysis with a phospho-specific antibody recognizing CREB on Ser133. LPA-induced phosphorylation of CREB was dependent on the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38 MAPK). Inhibition of ERK1/2 with PD98059 and of p38 MAPK with SB203580 efficiently blocked LPA-mediated phosphorylation of CREB. The LPA-induced CREB phosphorylation was abolished by H89, an inhibitor of mitogen- and stress-activated protein kinase-1 (MSK1). Together, these data suggest that LPA stimulates nuclear transcription factor CREB via mitogen-activated protein kinase signaling components, ERK1/2, p38 MAPK, and MSK1 in Rat-2 fibroblast cells.

Glial expression of the 90-kDa heat shock protein (HSP90) and the 94-kDa glucose-regulated protein (GRP94) following an excitotoxic lesion in the mouse hippocampus

Heat shock proteins (HSPs) are immediately expressed in neuronal and glial cells under various stressful conditions and play a protective role through molecular chaperones. Although several studies have been focused on the expression of HSPs, little is known about HSP90s expression in glial cells under neuropathological conditions. In this study, we evaluated the expression pattern of the glial cell‐related HSP90 and GRP94 proteins, following the induction of an excitotoxic lesion in the mouse brain. Adult mice received an intracerebroventricular injection of kainic acid; the brain tissue was then analyzed immunohistochemically for HSPs and double labeling using glial markers. HSPs expression was quantified by Western blot analysis. Excitotoxic damage was found to cause pyramidal cell degeneration in the CA3 region of the hippocampus. In the injured hippocampus, reactive microglia/macrophages expressed HSP90 from 12 h until 7 days postlesion (PL), showing maximal levels at day 1. In parallel, hippocampal reactive astrocytes showed the expression of GRP94 from 12 h until 7 days PL. In general, HSPs expression was transient, peaked at 1–3 days PL and reached basal levels by day 7. For the first time, our data demonstrate the injury‐induced expression of HSP90 and GRP94 in glial cells, which may contribute to the mechanism of glial cell protection and adaptation in response to damage, thereby playing an important role in the evolution of the glial response and the excitotoxic lesion outcome. HSP90 may provide antioxidant protective mechanisms against microglia/macrophages, whereas GRP94 may stabilize the astroglial cytoskeleton and participate in astroglial antioxidant mechanisms.

Increased expression of phosphatase and tensin homolog in reactive astrogliosis following intracerebroventricular kainic acid injection in mouse hippocampus.

A phosphatase and tensin homolog (PTEN) has been known to play multiple biological roles. However, role of PTEN in astrocyte activation is not clear yet. In the present study, the expression pattern of PTEN in the process of reactive gliosis was immunohistochemically examined in intracerebroventricular (i.c.v.) injected kainic acid mouse hippocampus. Mice were grouped into three; 30 min, 1 day and 7 days after kainic acid i.c.v. injection. Thirty minutes after kainic acid i.c.v. injection, astrocytes were activated and PTEN was weakly expressed in immature astrocytes. Seven days after kainic acid i.c.v. injection, PTEN expression was decreased in highly activated astrocytes showing extensively spindled shape. Immunofluorescence double labeling experiment showed that PTEN was expressed in glial fibrillary acidic protein-positive astrocytes. These findings suggest that PTEN might have a role in early stage of reactive astrogliosis in vivo.

Surgical resection of nodular ground-glass opacities without percutaneous needle aspiration or biopsy

BackgroundPercutaneous needle aspiration or biopsy (PCNA or PCNB) is an established diagnostic technique that has a high diagnostic yield. However, its role in the diagnosis of nodular ground-glass opacities (nGGOs) is controversial, and the necessity of preoperative histologic confirmation by PCNA or PCNB in nGGOs has not been well addressed.MethodsWe here evaluated the rates of malignancy and surgery-related complications, and the cost benefits of resecting nGGOs without prior tissue diagnosis when those nGGOs were highly suspected for malignancy based on their size, radiologic characteristics, and clinical courses. Patients who underwent surgical resection of nGGOs without preoperative tissue diagnosis from January 2009 to October 2013 were retrospectively analyzed.ResultsAmong 356 nGGOs of 324 patients, 330 (92.7%) nGGOs were resected without prior histologic confirmation. The rate of malignancy was 95.2% (314/330). In the multivariate analysis, larger size was found to be an independent predictor of malignancy (odds ratio, 1.086; 95% confidence interval, 1.001-1.178, p =0.047). A total of 324 (98.2%) nGGOs were resected by video-assisted thoracoscopic surgery (VATS), and the rate of surgery-related complications was 6.7% (22/330). All 16 nGGOs diagnosed as benign nodules were resected by VATS, and only one patient experienced postoperative complications (prolonged air leak). Direct surgical resection without tissue diagnosis significantly reduced the total costs, hospital stay, and waiting time to surgery.ConclusionsWith careful selection of nGGOs that are highly suspicious for malignancy, surgical resection of nGGOs without tissue diagnosis is recommended as it reduces costs and hospital stay.

Synthesis of oligosaccharide-containing orange juice using glucansucrase

Orange juice is a well-accepted fruit juice, and is a natural source of various vitamins, especially vitamin C, as well as sugar, potassium, thiamine, folate, flavonoids and antioxidants. The respective fructose, glucose, and sucrose concentrations were 9.3, 22.9, and 48.1 g/L in the original orange juice used in this study, and 183.4, 170.1, and 142.8 g/L after concentration. Over 97% of the sucrose in the juice was enzymatically converted to glucooligosaccharides upon addition of 3 U/mL dextransucrase, prepared from Leuconostoc mesenteroides 512FMCM, at 16°C. The synthesized oligosaccharides comprised 35.0% of the total saccharides in the concentrated juice and 31.7% in the original juice. The optimum conditions for oligosaccharide synthesis using the concentrated juice were 35.2 × 10−1 U/mL dextransucrase and 1% Ca(OH)2. The calories in the original and modified concentrated orange juices were 325.4 and 246.7 kcal/L, respectively. Compared to the original concentrated juice, the enzyme-modified concentrated juice prevented the formation of 62.7% of the insoluble glucan resulting from addition of mutansucrase, produced by Streptococcus mutans.

Long-Term Follow-up of Small Pulmonary Ground-Glass Nodules Stable for 3 Years: Implications of the Proper Follow-up Period and Risk Factors for Subsequent Growth

Introduction: How long persistent and stable ground‐glass nodules (GGNs) should be followed is uncertain, although a minimum of 3 years is suggested. Here, we evaluated a group of GGNs that had remained stable for an initial period of 3 years with the aim of determining the proportion of GGNs showing subsequent growth after the initial 3 years and identifying the clinical and radiologic factors associated with subsequent growth. Methods: We retrospectively analyzed patients who underwent further computed tomography (CT) after the initial 3‐year follow‐up period showing a persistent and stable GGN (at least 5 years of follow‐up from the initial CT). Results: Between May 2003 and June 2015, 453 GGNs (438 pure GGNs and 15 part‐solid GGNs) were found in 218 patients. Of the 218 patients, 14 had 15 GGNs showing subsequent growth after the initial 3 years during the median follow‐up period of 6.4 years. For the person‐based analysis, the frequency of subsequent growth of GGNs that had been stable during the initial 3 years was 6.7% (14 of 218). For the nodule‐based analysis, the frequency was 3.3% (15 of 453). In a multivariate analysis, age 65 years or older (OR = 5.51, p = 0.012), history of lung cancer (OR = 6.44, p = 0.006), initial size 8 mm or larger (OR = 5.74, p = 0.008), presence of a solid component (OR = 16.58, p = 0.009), and air bronchogram (OR = 5.83, p = 0.015) were independent risk factors for subsequent GGN growth. Conclusions: For the individuals with GGNs having the aforementioned risk factors, the longer follow‐up period is required to confirm subsequent GGN growth.

Gastroprotective and gastric motility benefits of AD-lico/Healthy Gut™ Glycyrrhiza inflata extract

ABSTRACT The aim of this study was to evaluate in vivo both the anti-Helicobacter and the gastric-relaxing effects of AD-lico/Healthy Gut™ in rat models. AD-lico/Healthy Gut™ is a specially prepared commercial formulation of Glycyrrhiza inflata extract that is under clinical development for indications of gastrointestinal disease and inflammatory bowel disease. In the current study, the oral administration of AD-lico/Healthy Gut™ significantly reduced mucosal damage from Helicobacter pylori in rats and decreased the expression of the inflammatory markers iNOS and COX-2 in the test cells. AD-lico/Healthy Gut™ also reduced mucosal damage caused by water immersion stress in rats. The accelerated gastric emptying in normal rats was also seen with AD-lico/Healthy Gut™, providing relief in gastric relaxation in the test animals. The special formulation of AD-lico/Healthy Gut™ with reduced levels of component glycyrrhizin also has benefits in minimizing the potential side effects attributed to glycyrrhizin seen with similar Glycyrrhiza extracts in terms of induction of hypokalemia and muscle weakness. The preparation has a relatively high phenolic compound content relative to other methods of preparation and is indicative of lower glycyrrhizin levels. These results suggest that AD-lico/Healthy Gut™ may provide the necessary relief from a number of stomach discomfort issues faced by a large population of people.

Gold Thread Acupuncture for a Headache

A 39-year-old Korean woman presented with chronic cough. She never smoked and had postnasal drip. After physical examination, plain chest and paranasal sinus x-rays were performed, and no structural abnormalities were observed. She was finally diagnosed with upper airway cough syndrome. However, plain radiographs of the paranasal sinuses revealed hundreds of gold threads in the head (Figure 1). She had previously suffered from a headache for several years and had undergone acupuncture with gold threads as an adjuvant therapy for her headache. A clinical examination did not reveal signs of local inflammation, such as swelling, redness or tenderness, and she did not complain of pain within the soft tissue of the face. No specific management was performed for the gold threads. Acupuncture with gold threads has been occasionally used for the management of pain such as headaches, osteoarthritis and facial rejuvenation in Asian countries as a complementary therapy. Insertion of thin pieces of sterile gold threads into painful regions is believed to result in continuous stimulation in the body and relief of pain; however, little evidence is available to support this. Although gold is an inert material, permanent implantation of foreign bodies may lead to unexpected complications resulting in morbidity. Several reports have suggested that the implanted gold threads can become fragmented and migrate. Stark and Bannasch reported the case of a middle-aged Russian woman who experienced severe pain in her face and neck areas after repeated gold thread facelifts for facial rejuvenation. Her skull radiographs revealed numerous fragmented gold threads whose vector orientation had been lost, thus making it difficult for them to function as mechanical suspension vectors for the facelifts. Although fragmentation and migration of the gold threads result in severe pain, complete surgical removal of these foreign bodies is impossible and the pain may intensify. Another complication is local inflammation. Formation of granulomas because of gold thread implantation has been reported. A Korean woman developed erythematous nodules with a purulent discharge 2 months after acupuncture with gold threads for a facelift. Biopsy showed acute and chronic inflammatory cells with foreign body–type giant cells, which led to the diagnosis of a foreign body reaction. Although she was successfully treated with incision, drainage and oral corticosteroids, foreign bodies can lead to secondary infections. Theoretically, fragmented and migratory gold threads can damage arteries and can interfere with the analysis of radiographic images, as they obscure areas of anatomical structures. To date, there is no scientific evidence to support the efficacy of acupuncture with gold threads. In most cases, patients are not fully informed of the relevant risks and other reasonable treatment options. Sometimes, this acupuncture is performed by an unqualified practitioner. Health care providers should provide patients with the appropriate information, and the patients should be informed of reasonable alternatives available to gold thread implantation and the related complications.

UV-B induced fibrillization of crystallin protein mixtures

Environmental factors, mainly oxidative stress and exposure to sunlight, induce the oxidation, cross-linking, cleavage, and deamination of crystallin proteins, resulting in their aggregation and, ultimately, cataract formation. Various denaturants have been used to initiate the aggregation of crystallin proteins in vitro. All of these regimens, however, are obviously far from replicating conditions that exist in vivo that lead to cataract formation. In fact, it is our supposition that only UV-B radiation may mimic the observed in vivo cause of crystallin alteration leading to cataract formation. This means of inducing cataract formation may provide the most appropriate in vitro platform for in-depth study of the fundamental cataractous fibril properties and allow for testing of possible treatment strategies. Herein, we showed that cataractous fibrils can be formed using UV-B radiation from α:β:γ crystallin protein mixtures. Characterization of the properties of formed aggregates confirmed the development of amyloid-like fibrils, which are in cross-β-pattern and possibly in anti-parallel β-sheet arrangement. Furthermore, we were also able to confirm that the presence of the molecular chaperone, α-crystallin, was able to inhibit fibril formation, as observed for ‘naturally’ occurring fibrils. Finally, the time-dependent fibrillation profile was found to be similar to the gradual formation of age-related nuclear cataracts. This data provided evidence for the initiation of fibril formation from physiologically relevant crystallin mixtures using UV-B radiation, and that the formed fibrils had several traits similar to that expected from cataracts developing in vivo.

Glucooligosaccharide production by Leuconostoc mesenteroides fermentation with efficient pH control, using a calcium hydroxide-sucrose solution

Abstract95.3% of the sucrose in a feed batch fermentation (300 g/L) was hydrolyzed by Leuconostoc mesenteroides subp. mesenteroides NRRL B-23188 glucansucrase. Further, the glucose of sucrose formed glucooligosaccharides (GOS) of degree of polymerization (DP) over 2, together with 91.6% of the maltose (200 g/L). Lime saccharate (lime sucrate) was used to control the pH during fermentation. The GOS products had DP between 2 and 7. When Streptococcus mutans mutansucrase (0.1 U/mL) reacted with 0.1% sucrose, addition of 0.1 ~ 10% GOS to the mutansucrase reaction digest resulted in a 56 ~ 90% reduction of mutan formation. GOS also reduced E. coli (72.2%) and Salmonella sp. (over 40.0%) growth, when 2.5% GOS was used as a single carbon source, compared to growth using glucose. The calculated glycemic index and glycemic load of GOS was 8 and 1, respectively, based on a 10 g carbohydrate serving. GOS was calculated to have 2.43 kcal/g. After a glucose tolerance test was performed using C57BL/6 mice, we found that mice treated with GOS showed a 59.4% lower increase in plasma glucose than those treated with maltose.