A newly identified glutaminase-free ʟ-asparaginase (ʟ-ASPG86) from the marine bacterium Mesoflavibacter zeaxanthinifaciens

ʟ-Asparaginase (EC 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the ʟ-asparaginase gene (ʟ-ASPG86) in the genus Mesoflavibacter, which consists of a 1035-bp open reading frame (ORF) encoding 344 amino acids (aa). Following phylogenetic analysis, the deduced amino acid sequence of ʟ-ASPG86 (ʟ-ASPG86) grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The ʟ-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant ʟ-asparaginase (r-ʟ-ASPG86) showed optimum conditions at 37-40oC, pH 9. Moreover, r-ʟ-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-ʟ-ASPG86 was 687.1 units/mg under optimum conditions (37oC, pH 9 and 5 mM MnSO4). Please click Additional Files below to see the full abstract

Review of child maltreatment prevention programs in South Korea

Objective: The objective of this study is twofold. The first objective is to examine the status of studies on the effect of child maltreatment prevention programs in South Korea. The second objective is to recommend directions for child maltreatment prevention programs based on the results of the first objective.Procedure: A literature search of prevention programs for child maltreatment was conducted. Research studies published from 1989 to 2013 were identified through a computerized selection process, resulting in 198 studies. Further detailed search of keywords ultimately identified 37 studies focusing on child maltreatment prevention programs. These 37 studies were further classified and reviewed. Results: This study was able to find several pertinent findings. First, child maltreatment prevention programs targeting teachers and parents were not sufficient. Second, child maltreatment prevention programs for high risk population were also insufficient in the selected literatures. Third, based on the existing studies, child maltreatment prevention programs in South Korea lacked diversified contents. Fourth, most research measured effectiveness through attitude and awareness factors. Finally, child maltreatment programs were not systematic and mainly were conducted as one-time programs.Conclusions: There needs to be more studies examining the effectiveness of prevention programs for child maltreatment, notably, sustainable and long-term effects of prevention programs. Moreover, these programs need to include high-risk populations, with more diverse contents, that are systematically included throughout all three levels of prevention – primary, secondary, and tertiary

IFN-gamma signaling in the central nervous system controls the course of experimental autoimmune encephalomyelitis independently of the localization and composition of inflammatory foci

<p>Abstract</p> <p>Background</p> <p>Murine experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, presents typically as ascending paralysis. However, in mice in which interferon-gamma (IFNγ) signaling is disrupted by genetic deletion, limb paralysis is accompanied by atypical deficits, including head tilt, postural imbalance, and circling, consistent with cerebellar/vestibular dysfunction. This was previously attributed to intense cerebellar and brainstem infiltration by peripheral immune cells and formation of neutrophil-rich foci within the CNS. However, the exact mechanism by which IFNγ signaling prohibits the development of vestibular deficits, and whether the distribution and composition of inflammatory foci within the CNS affects the course of atypical EAE remains elusive.</p> <p>Methods</p> <p>We induced EAE in IFNγ-/- mice and bone marrow chimeric mice in which IFNγR is not expressed in the CNS but is intact in the periphery (IFNγR^CNSKO) and vice versa (IFNγR^periKO). Blood-brain barrier permeability was determined by Evans blue intravenous administration at disease onset. Populations of immune cell subsets in the periphery and the CNS were quantified by flow cytometry. CNS tissues isolated at various time points after EAE induction, were analyzed by immunohistochemistry for composition of inflammatory foci and patterns of axonal degeneration.</p> <p>Results</p> <p>Incidence and severity of atypical EAE were more pronounced in IFNγR^CNSKO as compared to IFNγR^periKO mice. Contrary to what we anticipated, cerebella/brainstems of IFNγR^CNSKO mice were only minimally infiltrated, while the same areas of IFNγR^periKO mice were extensively populated by peripheral immune cells. Furthermore, the CNS of IFNγR^periKO mice was characterized by persistent neutrophil-rich foci as compared to IFNγR^CNSKO. Immunohistochemical analysis of the CNS of IFNγ-/- and IFNγR chimeric mice revealed that IFNγ protective actions are exerted through microglial STAT1.</p> <p>Conclusions</p> <p>Alterations in distribution and composition of CNS inflammatory foci are not sufficient for the onset of atypical EAE. IFNγ dictates the course of neuroinflammatory disorders mainly through actions exerted within the CNS. This study provides strong evidence that link microglial STAT1 inactivation to vestibular dysfunction.</p

Biochemical properties of a novel neoagarotriose-producing β-agarase from Gilvimarinus agarolyticus JEA5

An agar degrading bacterium was isolated from seawater, collected from the east coast of Jeju Island, republic of Korea and identified as Gilvimarinus agarolyticus JEA5. The β-agarase gene from Gilvimarinus agarolyticus JEA5 (rGaa16B) was identified from draft genome sequence by BLAST. Gaa16B has 1800 bp of open reading frame encoding 636 amino acids (aa), and include glycosyl hydrolase family 16 (GH16) β-agarase module and two carbohydrate binding module 6 (CBM6). The Gaa16b was cloned and overexpressed as a MBP-fusion recombinant β-agarase (without signal peptide and two CBM6) in E. coil. rGaa16B showed highest activity at 60°C and pH 7. After incubation at 45OC for 90 min, rGaa16B showed over than 95% of its initial activity. rGaa16B were enhanced in the presence of MnCl2, KCl2, MgCl2, FeSO4. rGaa16B showed 2112.1 unit/mg in the presence of 2.5 mM of MnCl2. rGaa16B produce mainly neoagartetraose (NA4) and neoagarobiose (NA2). Interestingly, we observed neoagartriose (NA3) from hydrolytic products of rGaa16B. LC/Mass analysis was performed to confirm the hydrolytic products containing neoagarotriose. We found three different hydrolytic products which showed 324.28, 468.41, 630.55 Da of molecular weight, respectively. Please click Additional Files below to see the full abstract

Molecular cloning and Biochemical properties of GH-16 β-agarase from Gilvimarinus agarolyticus JEA5

Agar is complex polysaccharide founds in the cell walls of some red algae and up to 70 % of the algal cell wall can be agar polymers. Agar was formed by a mixture of two polysaccharides named agarose and agaropectin. Agarose can be hydrolyzed by α-agarase (E.C. 3.2.1.158) and by β-agarase (E.C. 3.2.1.81); the former cleaves the α-1, 3 linkage of agarose to generate agaro-oligosaccharides, and the latter cleaves the β-1,4 linkage to generate neoagaro-oligosaccharides. Agarases have been isolated from many sources, including seawater, marine sediments, marine algae, marine mollusks, fresh water and soil. Recently, Givimarinus chinensis, G. polysacchalyticus, G. agarilyticus were identified and their agarolytic activity also reported. However, there are no report published that molecular and functional characterization of agarase from Givimarinus genus. In this study, we first report molecular characterization and biochemical properties of agarase from Gilvimarinus genus. Please click Additional Files below to see the full abstract

An infant case of ruptured umbilical hernia with eviscerated omentum after acupuncture

Serious acupuncture-related complications are rare in infants. We experienced a case of a 58-day-old infant with an umbilical hernia who received acupuncture from a non-professional guardian. Upon initial examination in the emergency department, the omentum protruded through the skin defect caused by acupuncture performed on top of the hernia. The infant underwent emergency surgery, and subsequently recovered uneventfully

Intakes of Dairy Products and Calcium and Obesity in Korean Adults: Korean National Health and Nutrition Examination Surveys (KNHANES) 2007-2009

Background: The possible effects of dairy product intake against obesity have been suggested in animal studies; however, the association is still not well established in epidemiological studies. Few studies in Asian countries with relatively low intake of dairy products exist. Objective: We investigated the association between dairy products and calcium intake and obesity in Korean population with relatively low intake of dairy products. Subjects and Methods Our study population consisted of adults (n = 7173) aged 19–64 among participants of the 2007, 2008 and 2009 Korean National Health and Nutrition Examination Survey who had not made any attempt of intentional weight loss. Dietary intake data from food frequency questionnaire (FFQ) and 24-hour recall were used. Dairy products included milk and yogurt in the FFQ. Obesity was defined as BMI≥25 kg/m2. Results: Higher frequency of dairy product intake was associated with a reduced prevalence of obesity (OR = 0.63; 95% CI = 0.45–0.89 for ≥2 times/day vs. ≤1 time/month; p for trend = 0.003) using the intake data from FFQ. Similarly, high frequency of milk or yogurt intake had an inverse association with obesity. The association between milk and yogurt intake and obesity was similar when the intake from 24-hour recall was examined. Higher calcium intake from dairy products as well as total dietary calcium intake was associated with a decreased prevalence of obesity (OR = 0.83; 95% CI = 0.71–0.98 for highest vs. lowest quintile of dairy calcium intake; p for trend = 0.02, OR = 0.78; 95% CI = 0.64–0.94 for highest vs. lowest quintile of total calcium intake; p for trend = 0.04). The associations appeared to be stronger in women than in men. Conclusion: These results suggest that high consumption of dairy products is associated with a lower prevalence of obesity and that calcium in dairy products may be one of the components contributing to the association. Further longitudinal studies are warranted to replicate our findings