Epac2a-null mice exhibit obesity-prone nature more susceptible to leptin resistance.

BACKGROUND: The exchange protein directly activated by cAMP (Epac), which is primarily involved in cAMP signaling, has been known to be essential for controlling body energy metabolism. Epac has two isoforms: Epac1 and Epac2. The function of Epac1 on obesity was unveiled using Epac1 knockout (KO) mice. However, the role of Epac2 in obesity remains unclear. METHODS: To evaluate the role of Epac2 in obesity, we used Epac2a KO mice, which is dominantly expressed in neurons and endocrine tissues. Physiological factors related to obesity were analyzed: body weight, fat mass, food intake, plasma leptin and adiponectin levels, energy expenditure, glucose tolerance, and insulin and leptin resistance. To determine the mechanism of Epac2a, mice received exogenous leptin and then hypothalamic leptin signaling was analyzed. RESULTS: Epac2a KO mice appeared to have normal glucose tolerance and insulin sensitivity until 12 weeks of age, but an early onset increase of plasma leptin levels and decrease of plasma adiponectin levels compared with wild-type mice. Acute leptin injection revealed impaired hypothalamic leptin signaling in KO mice. Consistently, KO mice fed a high-fat diet (HFD) were significantly obese, presenting greater food intake and lower energy expenditure. HFD-fed KO mice were also characterized by greater impairment of hypothalamic leptin signaling and by weaker leptin-induced decrease in food consumption compared with HFD-fed wild-type mice. In wild-type mice, acute exogenous leptin injection or chronic HFD feeding tended to induce hypothalamic Epac2a expression. CONCLUSIONS: Considering that HFD is an inducer of hypothalamic leptin resistance and that Epac2a functions in pancreatic beta cells during demands of greater work load, hypothalamic Epac2a may have a role in facilitating leptin signaling, at least in response to higher metabolic demands. Thus, our data indicate that Epac2a is critical for preventing obesity and thus Epac2a activators may be used to manage obesity and obesity-mediated metabolic disorders.

Effects of Allopurinol and Apocynin on Renal Ischemia-Reperfusion Injury in Rats.

BACKGROUND: This study evaluated the effects of allopurinol (ALP), a xanthine oxidase inhibitor, and apocynin (APC), a NADPH oxidase inhibitor, administered alone or together, on kidney damage caused by renal ischemia-reperfusion (IR) in rats. METHODS: Thirty rats were randomly assigned to 5 groups. Group 1 was a sham group. Group 2 was the renal IR control group (30-min ischemia followed by 24-h reperfusion). In groups 3 and 4, ALP or APC, respectively, was administered 1 h before the ischemia. In group 5, ALP and APC were co-administered. Blood urea nitrogen (BUN) and serum creatinine (Cr), renal tissue malondialdehyde (MDA) and superoxide dismutase (SOD), and histological changes were evaluated. RESULTS: A significant increase in BUN and Cr level, and histological damage was seen in the IR control group, indicating renal injury. Elevated MDA and decreased SOD levels in the IR control group demonstrated that renal damage occurred through oxidative stress. Pretreatment with ALP or APC alone or together prevented IR-induced renal damage. However, there was no significant difference between treatment with a single drug and co-administration of ALP and APC. CONCLUSIONS: The use of ALP and/or APC before ischemia may be beneficial to ameliorate renal IR injury.

First Report of Pectobacterium carotovorum subsp. carotovorum Causing Soft Rot of Orostachys japonica in Korea.

Orostachys japonica (Maxim) A. Berger is an important traditional medicine in Korea. The extract of this plant has antioxidant activity and suppresses cancer cell proliferation (1). From summer through fall of 2012 and 2013, a high incidence (~10% to 30%) of disease outbreaks of all plants characterized by water-soaked lesions and soft rot with a stinky odor was observed in cultivated O. japonica around Uljin (36°59'35.04″N, 126°24'1.51″E), Korea. Water-soaked lesions were first observed on the stem base of plants. Subsequently, the plants collapsed, although the upper portion remained asymptomatic. Thereafter, the lesions expanded rapidly over the entire plant. To isolate potential pathogens from infected leaves, small sections (5 to 10 mm2) were excised from the margins of lesions. Ten bacteria were isolated from ten symptomatic plants. Three representative isolates from different symptomatic plants were used for identification and pathogenicity tests. Isolated bacteria were gram negative, pectolytic on crystal violet pectate agar, nonfluorescent on King's medium B, and elicited a hypersensitive response in tobacco plants. All isolates caused soft rot of potato tubers. These isolates also differed from isolates of Erwinia chrysanthemi (Ech) that they were insensitive to erythromycin and did not produce phosphatase. These isolates differed from known strains of E. carotovora subsp. atroseptica in that they did not produce reducing substances from sucrose (2). Use of the Biolog GN microplate and the Release 4.0 system identified the isolate as Pectobacterium carotovorum subsp. carotovorum with 81.2% similarity. The 16S rRNA of the isolated bacteria was amplified by PCR and sequenced as described by Weisburg et al. (3). A BLAST analysis for sequence similarity of the 16S rRNA region revealed 99% similarity with nucleotide sequences for P. carotovorum subsp. carotovorum isolates (KC790305, KC790280, JF926758, JX196705, and AB680074). The pathogenicity of three bacterial isolates was examined on three 2-year-old O. japonica plants by adding 50 µl of a bacterial suspension containing 108 CFU/ml when wounding the leaves with sterile needles. Ten control plants were inoculated with sterilized water. After inoculation, plants were maintained in a growth chamber at 25°C with relative humidity ranging from 80 to 90%. After 2 to 3 days, tissue discoloration, water-soaked lesions, and soft rot developed around the inoculation point. Severe symptoms of soft rot and darkening developed on leaves of inoculated plants within 3 to 5 days after inoculation. All controls remained healthy during these experiments. The bacterial strains re-isolated from the parts of the leaf showing the symptoms and identified as P. carotovorum subsp. carotovorum on the basis of the biochemical and physiological tests, as well as Biolog system. The results obtained for pathogenicity, Biolog analysis, and molecular data corresponded with those for P. carotovorum subsp. carotovorum. To our knowledge, this is the first report of the presence of P. carotovorum on O. japonica in Korea. References: (1) C.-H. Kim et al. Kor. J. Med. Crop Sci. 11:31, 2003. (2) N. W. Schaad et al. Erwinia Soft Rot Group. Page 56 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al. eds. American Phytopathological Society, St. Paul. MN, 2001. (3) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991.

Enhanced anti-tumor effects of combined MDR1 RNA interference and human sodium/iodide symporter (NIS) radioiodine gene therapy using an adenoviral system in a colon cancer model.

Using an adenoviral system as a delivery mediator of therapeutic gene, we investigated the therapeutic effects of the use of combined MDR1 shRNA and human NIS (hNIS) radioiodine gene therapy in a mouse colon xenograft model. In vitro uptake of Tc-99m sestamibi was increased approximately two-fold in cells infected with an adenovirus vector that expressed MDR1 shRNA (Ad-shMDR1) and I-125 uptake was 25-fold higher in cells infected with an adenovirus vector that expressed human NIS (Ad-hNIS) as compared with control cells. As compared with doxorubicin or I-131 treatment alone, the combination of doxorubicin and I-131 resulted in enhanced cytotoxicity for both Ad-shMDR1- and Ad-hNIS-infected cells, but not for control cells. In vivo uptake of Tc-99m sestamibi and Tc-99m pertechnetate was twofold and 10-fold higher for Ad-shMDR1 and Ad-hNIS-infected tumors as compared with tumors infected with a control adenovirus construct that expressed beta-galactosidase (Ad-LacZ), respectively. In mice treated with either doxorubicin or I-131 alone, there was a slight delay in tumor growth as compared to mice treated with Ad-LacZ. However, combination therapy with doxorubicin and I-131 induced further significant inhibition of tumor growth as compared with mice treated with Ad-LacZ. We have shown successful therapeutic efficacy of combined MDR shRNA and hNIS radioiodine gene therapy using an adenoviral vector system in a mouse colon cancer model. Adenovirus-mediated cancer gene therapy using MDR1 shRNA and hNIS would be a useful tool for the treatment of cancer cells expressing multi-drug resistant genes.

Effects of sevoflurane on collagen production and growth factor expression in rats with an excision wound.

BACKGROUND: Sevoflurane is a widely used inhalation anesthetic, but there are no studies on its effect on the wound-healing process. This study was undertaken to evaluate the effect of exposure time to sevoflurane on wound healing. METHOD: Male Sprague-Dawley rats were used. Two circular full-thickness skin defects 8 mm in diameter were made on the dorsum of the rats. The animals were divided into six groups according to exposed gas type and time: S1 (sevoflurane, 1 h), S4 (sevoflurane, 4 h), S8 (sevoflurane, 8 h), O1 (oxygen, 1 h), O4 (oxygen, 4 h), and O8 (oxygen, 8 h). The surface area of the wounds was measured 0, 1, 3, and 7 days after surgery. Separately, the mean blood pressures (MBP) and arterial oxygen pressures (PaO(2)) were monitored during the sevoflurane exposure. Collagen type I production and transforming growth factor-beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) expression on the wound surface were analyzed. Routine histological analysis was also performed. RESULT: Exposure duration to sevoflurane had no influence on MBP and PaO(2). The reduction in wound size and collagen type I production was delayed in S8. The expression of TGF-beta1 and bFGF on the wound surface in S8 was significantly attenuated in S8. The histology of the S8 demonstrated a delayed healing status. CONCLUSIONS: Prolonged exposure to sevoflurane might alter the inflammatory phase of the wound-healing process by attenuation of growth factor expression such as TGF-beta1 and bFGF and subsequently by reduced collagen production.

Multiple hepatic hemangiomas with fluid-fluid levels.

Hepatic haemangiomas with fluid-fluid levels are known to be rare with only five cases reported in the English literature. According to the previous reports, the presence of fluid-fluid level could attribute to the separation of blood cells and serous fluid because of the extremely slow flow in cavernous haemangioma of the liver. We describe the imaging features of multiple hepatic haemangiomas with fluid-fluid levels, which was pathologically proven with ultrasonography guided biopsy.

Acetic acid as a sclerosing agent for renal cysts: comparison with ethanol in follow-up results.

PURPOSE: To compare follow-up results of sclerotherapy for renal cyst using 50% acetic acid with those using 99% ethanol as sclerosing agents. METHODS: Eighty-one patients underwent sclerotherapy and 58 patients, 23 males, 35 females, aged 6-76 years, having a total of 60 cysts, were included in this study; the others were lost to follow-up. The renal cysts were diagnosed by sonography, computed tomography (CT), or magnetic resonance imaging (MRI). Sclerotherapy was performed using 50% acetic acid for 32 cysts in 31 patients and 99% ethanol for 28 cysts in 27 patients. Under fluoroscopic guidance, cystic fluid was aspirated as completely as possible. After instillation of a sclerosing agent corresponding to 11.7%-25% (4-100 ml) of the aspirated volume, the patient changed position for 20 min and then the agent was removed. Patients were followed up by sonography for a period of 1-49 months. The volume of the renal cyst after sclerotherapy was compared with that of the renal cyst calculated before sclerotherapy. Medical records were reviewed to analyze complications. RESULTS: The mean volume after sclerotherapy of the 17 cysts followed for 3-4 months in the acetic acid group was 5.1% of the initial volume, and for the 14 cysts in the ethanol group it was 10.2%. Complete regression during follow-up was shown in 21 cysts (66%) in the acetic acid group; the mean volume of these cysts before the procedure was 245 ml. The mean volume of the nine (32%) completely regressed cysts in the ethanol group was 184 ml. Mild flank pain, which occurred in three patients in each group, was the only complication and resolved the next day. CONCLUSION: Acetic acid was an effective and safe sclerosing agent for renal cysts, tending to induce faster and more complete regression than ethanol.

A molecular epidemiologic study of methicillin-resistant Staphylococcus aureus infection in patients undergoing middle ear surgery.

The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections after middle ear surgery has recently increased at our hospital. Most of these infections were thought to be hospital-acquired when medical personnel in contact with an MRSA-infected patient may have inadvertently transmitted the pathogen to other patients. To prevent further transmission it is essential that such sources of MRSA infection and transmission routes be selected out and eradicated. Therefore, it is necessary to determine whether the strains of MRSA isolated from infected patients are identical to those obtained from medical personnel in order to prove a reciprocal transmission of organisms between medical personnel and patients. Surveillance bacterial cultures from the anterior nares and hands of medical personnel working in the Department of Otolaryngology, Korea University Guro Hospital, were performed at two different time points: 6 December 1994 and 17 June 1996. Ribotyping with Southern blot technique was used to compare 12 MRSA strains from medical carriers with 60 strains identified from the otorrhea of MRSA-infected patients undergoing middle ear surgery. As results, six different MRSA strains were identified (types I, II, III, IV, V and VI) from ribotyping with EcoR1. One distinct subtype, type I strain, was the most frequently identified strain in both medical carriers and patients. Results also showed that 6 MRSA isolates from 10 medical carriers and 20 from 30 patients contained type I ribotype at first culture. Two medical carriers' isolates and 13 isolates from 30 patients shared the same type I strain at the second surveillance culture. In all, 41 out of 72 MRSA strains (56.9%) shared an identical ribotype pattern. Postoperative MRSA infection rates after treatment of medical carriers and the application of rigorous preventive procedures decreased from 11.9 to 5.7% after first culture and 9.0 to 7.7% following second cultures. These findings confirm that MRSA transmission can occur between medical personnel and patients and that effective preventive measures can reduce the postoperative infection rate.

Molecular characterization of a cDNA encoding chloroplastic fructose-1,6-bisphosphatase from soybean (Glycine max L.).

A full-length cDNA of soybean chloroplastic fructose-1,6-bisphosphatase was cloned and sequenced. The cDNA contained 1321 bp with 5' (26 bp) and 3' (88 bp) untranslated regions. The open reading frame of the cDNA contained 1206 bp corresponding to a polypeptide of 402 amino acids with 50 amino acid residues of a transit peptide at N-terminus that is necessary for transport into the chloroplast. A unique site relevant to the action of thioredoxin f was conserved at 221 amino acid residue. Northern blot analysis indicated that the expression of the enzyme was regulated by light illumination.