Proteome-Level Responses of Escherichia coli to Long-Chain Fatty Acids and Use of Fatty Acid Inducible Promoter in Protein Production

In Escherichia coli, a long-chain acyl-CoA is a regulatory signal that modulates gene expression through its binding to a transcription factor FadR. In this study, comparative proteomic analysis of E. coli in the presence of glucose and oleic acid was performed to understand cell physiology in response to oleic acid. Among total of 52 proteins showing altered expression levels with oleic acid presence, 9 proteins including AldA, Cdd, FadA, FadB, FadL, MalE, RbsB, Udp, and YccU were newly synthesized. Among the genes that were induced by oleic acid, the promoter of the aldA gene was used for the production of a green fluorescent protein (GFP). Analysis of fluorescence intensities and confocal microscopic images revealed that soluble GFP was highly expressed under the control of the aldA promoter. These results suggest that proteomics is playing an important role not only in biological research but also in various biotechnological applications

Transcriptome and proteome analyses of adaptive responses to methyl methanesulfonate in Escherichia coli K-12 and ada mutant strains

<p>Abstract</p> <p>Background</p> <p>The Ada-dependent adaptive response system in <it>Escherichia coli </it>is important for increasing resistance to alkylation damage. However, the global transcriptional and translational changes during this response have not been reported. Here we present time-dependent global gene and protein expression profiles following treatment with methyl methanesulfonate (MMS) in <it>E. coli </it>W3110 and its <it>ada </it>mutant strains.</p> <p>Results</p> <p>Transcriptome profiling showed that 1138 and 2177 genes were differentially expressed in response to MMS treatment in the wild-type and mutant strains, respectively. A total of 81 protein spots representing 76 nonredundant proteins differentially expressed were identified using 2-DE and LC-MS/MS. In the wild-type strain, many genes were differentially expressed upon long-exposure to MMS, due to both adaptive responses and stationary phase responses. In the <it>ada </it>mutant strain, the genes involved in DNA replication, recombination, modification and repair were up-regulated 0.5 h after MMS treatment, indicating its connection to the SOS and other DNA repair systems. Interestingly, expression of the genes involved in flagellar biosynthesis, chemotaxis, and two-component regulatory systems related to drug or antibiotic resistance, was found to be controlled by Ada.</p> <p>Conclusion</p> <p>These results show in detail the regulatory components and pathways controlling adaptive response and how the related genes including the Ada regulon are expressed with this response.</p

Discovery of Novel Sources of Vitamin B12 in Traditional Korean Foods from Nutritional Surveys of Centenarians

Human longevity can be explained by a variety of factors, among them, nutritional factor would play an important role. In our study of Korean centenarians for their longevity, the apparent nutritional imbalance in the traditional semi-vegetarian diet raised a special attention, especially on vitamin B12 status, supplied by animal foods. Interestingly, we found that the prevalence of vitamin B12 deficient Korean centenarians was not higher compared with those from Western nations with animal-oriented traditional foods. We assumed that there might be some unveiled sources for vitamin B12 in the Korean traditional foods. Screening of vitamin B12 contents has revealed that some traditional soybean-fermented foods, such as Doenjang and Chunggukjang, and seaweeds contain considerable amounts of vitamin B12. Taken together, it can be summarized that the traditional foods, especially of fermentation, might be evaluated for compensation of the nutritional imbalance in the vegetable-oriented dietary pattern by supplying vitamin B12, resulting in maintenance of health status

Estimation of axial curvature of anterior sclera: correlation between axial length and anterior scleral curvature as affected by angle kappa

Background: Though the development and fitting of scleral contact lenses are expanding steadily, there is no simple method to provide scleral metrics for scleral contact lens fitting yet. The aim of this study was to establish formulae for estimation of the axial radius of curvature (ARC) of the anterior sclera using ocular biometric parameters that can be easily obtained with conventional devices. Methods: A semi-automated stitching method and a computational analysis tool for calculating ARC were developed by using the ImageJ and MATLAB software. The ARC of all the ocular surface points were analyzed from the composite horizontal cross-sectional images of the right eyes of 24 volunteers; these measurements were obtained using anterior segment optical coherence tomography for a previous study (AS-OCT; Visante). Ocular biometric parameters were obtained from the same volunteers with slit-scanning topography and partial coherence interferometry. Correlation analysis was performed between the ARC at 8 mm to the axis line (ARC[8]) and other ocular parameters (including age). With ARC obtained on several nasal and temporal points (7.0, 7.5, 8.0, 8.5, and 9.0 mm from the axis line), univariate and multivariate linear regression analyses were performed to develop a model for estimating ARC with the help of ocular biometric parameters. Results: Axial length, spherical equivalent, and angle kappa showed correlations with temporal ARC[8] (tARC[8]; Pearson’s r = 0.653, −0.579, and −0.341; P = 0.001, 0.015, and 0.015, respectively). White-to-white corneal diameter (WTW) and anterior chamber depth (ACD) showed correlation with nasal ARC[8] (nARC[8]; Pearson’s r = −0.492 and −0.461; P = 0.015 and 0.023, respectively). The formulae for estimating scleral curvatures (tARC, nARC, and average ARC) were developed as a function of axial length, ACD, WTW, and distance from the axis line, with good determinant power (72 − 80 %; SPSS ver. 22.0). Angle kappa showed strong correlation with axial length (Pearson’s r = −0.813, P <0.001), and the different correlation patterns of nasal and temporal ARC with axial length can be explained by the ocular surface deviation represented by angle kappa. Conclusions: Axial length, ACD, and WTW are useful parameters for estimating the ARC of the anterior sclera, which is important for the haptic design of scleral contact lenses. Angle kappa affects the discrepancies between the nasal and temporal scleral curvature.Korea (South). Ministry of Health & Welfare (Projects A084496 and A120018

TOLL-LIKE RECEPTOR 2 AND MUC2 EXPRESSION ON HUMAN INTESTINAL EPITHELIAL CELLS BY GYMNOPHALLOIDES SEOI ADULT ANTIGEN

Goblet cell hyperplasia and mucin hypersecretion are important for the expulsion of the intestinal trematode, Gymnophalloides seoi, from mice. However, regulatory mechanisms underlying these processes remain elusive. To better understand the effects of G. seoi antigen on the host`s intestinal epithelial cells, we determined whether G. seoi induces expression of Toll-like receptors (TLRs) and in mucin-related (MUC) genes on a human intestinal epithelial cell line (HT29 cells). We treated HT29 cells with G. senior other adult helminth antigens and measured mRNAs of TLRs and MUCs. We also performed reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry to determine whether TLR and MUC expression is regulated by interferon (IFN)-gamma, interleukin-4, or monoclonal antibodies (mAbs) against G. seoi 46 kDa antigen. Gymnophalloides seoi antigen significantly induced expression of TLR2 and MUC2 in HT29 cells, and IFN-gamma was found to upregulate TLR2 expression on the surface of the cells. The expression of MUC2 was increased by IFN-gamma, but was decreased significantly via the combination of mAbs-to-human TLRs and G. seal antigen. These results demonstrated that G. seoi antigen upregulates TLR2 and MUC2 expression on human intestinal epithelial cells. These effects reflect a helminth-induced. IFN-gamma dependent, and innate mucosal immune mechanism in this human intestinal cell line.Guk SM, 2009, J PARASITOL, V95, P581, DOI 10.1645/GE-1807.1Ueno K, 2008, AM J RESP CELL MOL, V38, P263, DOI 10.1165/rcmb.2007-0336RCKraft M, 2008, EUR RESPIR J, V31, P43, DOI 10.1183/09031936.00103307Andrianifahanana M, 2007, ONCOGENE, V26, P7251, DOI 10.1038/sj.onc.1210532Ikeda H, 2007, LAB INVEST, V87, P559, DOI 10.1038/labinvest.3700556Mueller T, 2006, J IMMUNOL, V176, P5805Harris G, 2006, WORLD J GASTROENTERO, V12, P2149Yamauchi J, 2006, APMIS, V114, P270, DOI 10.1111/j.1600-0463.2006.apm_353.xSchroder K, 2006, IMMUNOBIOLOGY, V211, P511, DOI 10.1016/j.imbio.2006.05.007Chen XM, 2005, J IMMUNOL, V175, P7447Ding SZ, 2005, HELICOBACTER, V10, P193Campos-Rodriguezp R, 2005, PARASITE IMMUNOL, V27, P1Akira S, 2004, NAT REV IMMUNOL, V4, P499, DOI 10.1038/nri1391Cario E, 2004, GASTROENTEROLOGY, V127, P224, DOI 10.1053/j.gastro.2004.04.015KAMMANADIMINTI SJ, 2004, FASEB J, V18, P155Seo M, 2003, J PARASITOL, V89, P1080, DOI 10.1645/GE-3182RNMoncada DM, 2003, TRENDS PARASITOL, V19, P305, DOI 10.1016/S1471-4922(03)00122-3McGuinness DH, 2003, TRENDS PARASITOL, V19, P312, DOI 10.1016/S1471-4922(03)00123-5Chai JY, 2003, TRENDS PARASITOL, V19, P109, DOI 10.1016/S1471-4922(02)00068-5Reddy PK, 2003, EUR J CANCER, V39, P397Coelho PS, 2002, J LEUKOCYTE BIOL, V71, P837Rose MC, 2001, AM J RESP CELL MOL, V25, P533Shekels LL, 2001, DIGEST DIS SCI, V46, P1757, DOI 10.1023/A:1010622125040Gouyer V, 2001, BBA-MOL CELL RES, V1539, P71DEPLANCKE B, 2001, AM J CLIN NUTR, V73, P1131CHAI JY, 2001, KOREAN J PARASITOL, V39, P23Enss ML, 2000, INFLAMM RES, V49, P162ONAH DN, 2000, KOREAN J PARASITOLOG, V38, P209Dabbagh K, 1999, J IMMUNOL, V162, P6233Cohn L, 1999, J IMMUNOL, V162, P6178LEE SH, 1994, AM J TROP MED HYG, V51, P281NAWA Y, 1994, PARASITE IMMUNOL, V16, P333LEE SH, 1993, J PARASITOL, V79, P677CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P159BEAVER PC, 1984, CLIN PARASITOLOGY

Multiple domains of TonEBP cooperate to stimulate transcription in response to hypertonicity.

Tonicity-responsive enhancer binding protein (TonEBP), also known as NFAT5, belongs to the Rel family of transcriptional activators. In the kidney medulla and thymus, TonEBP plays a major role in protecting renal cells and T cells from the deleterious effects of ambient hypertonicity. TonEBP is stimulated by hypertonicity via several pathways: increased expression of protein, nuclear translocation, and increased transactivation. In this study, we identified five domains of TonEBP involved in transactivation. The two conserved glutamine repeats were not involved in transactivation. There were three activation domains that could stimulate transcription independently. In addition, there were two modulation domains that potentiated the activity of the activation domains. One of the activation domains is unique to a splice isoform that is more active than others, indicating that alternative splicing can affect the activity of TonEBP. Another activation domain and one of the modulation domains were stimulated by hypertonicity. All the five domains acted in synergy in every combination. Although overall phosphorylation of TonEBP increased in response to hypertonicity, phosphorylation of the activation and modulation domains did not increase in isolation. In sum, TonEBP possesses far more elaborate domains involved in transactivation compared with other Rel proteins

Effect of GCSB-5, a Herbal Formulation, on Monosodium Iodoacetate-Induced Osteoarthritis in Rats

Therapeutic effects of GCSB-5 on osteoarthritis were measured by the amount of glycosaminoglycan in rabbit articular cartilage explants in vitro, in experimental osteoarthritis induced by intra-articular injection of monoiodoacetate in rats in vivo. GCSB-5 was orally administered for 28 days. In vitro, GCSB-5 inhibited proteoglycan degradation. GCSB-5 significantly suppressed the histological changes in monoiodoacetate-induced osteoarthritis. Matrix metalloproteinase (MMP) activity, as well as, the levels of serum tumor necrosis factor-α, cyclooxygenase-2, inducible nitric oxide synthase protein, and mRNA expressions were attenuated by GCSB-5, whereas the level of interleukin-10 was potentiated. By GCSB-5, the level of nuclear factor-κB p65 protein expression was significantly attenuated but, on the other hand, the level of inhibitor of κB-α protein expression was increased. These results indicate that GCSB-5 is a potential therapeutic agent for the protection of articular cartilage against progression of osteoarthritis through inhibition of MMPs activity, inflammatory mediators, and NF-κB activation

Experience of Comamonas Acidovorans Keratitis with Delayed Onset and Treatment Response in Immunocompromised Cornea

summary:We make some comments on the problem of how the Henstock-Kurzweil integral extends the McShane integral for vector-valued functions from the descriptive point of view