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Journal articles on the topic 'Lyk3'

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Published: 4 June 2021

Last updated: 7 February 2022

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Xue, De-Xing, Chun-Lian Li, Zhi-Ping Xie, and Christian Staehelin. "LYK4 is a component of a tripartite chitin receptor complex in Arabidopsis thaliana." Journal of Experimental Botany 70, no. 19 (2019): 5507–16. http://dx.doi.org/10.1093/jxb/erz313.

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LYK1, LYK4, and LYK5 form a tripartite receptor complex in Arabidopsis to perceive chitin, with constitutive LYK4–LYK5 and chitin-induced LYK1–LYK5 ectodomain interactions, and LYK4 functioning as a LYK5-associated co-receptor or scaffold protein.

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Liang, Pengbo, Thomas F. Stratil, Claudia Popp, et al. "Symbiotic root infections in Medicago truncatula require remorin-mediated receptor stabilization in membrane nanodomains." Proceedings of the National Academy of Sciences 115, no. 20 (2018): 5289–94. http://dx.doi.org/10.1073/pnas.1721868115.

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Plant cell infection is tightly controlled by cell surface receptor-like kinases (RLKs). Like other RLKs, the Medicago truncatula entry receptor LYK3 laterally segregates into membrane nanodomains in a stimulus-dependent manner. Although nanodomain localization arises as a generic feature of plant membrane proteins, the molecular mechanisms underlying such dynamic transitions and their functional relevance have remained poorly understood. Here we demonstrate that actin and the flotillin protein FLOT4 form the primary and indispensable core of a specific nanodomain. Infection-dependent induction of the remorin protein and secondary molecular scaffold SYMREM1 results in subsequent recruitment of ligand-activated LYK3 and its stabilization within these membrane subcompartments. Reciprocally, the majority of this LYK3 receptor pool is destabilized at the plasma membrane and undergoes rapid endocytosis in symrem1 mutants on rhizobial inoculation, resulting in premature abortion of host cell infections. These data reveal that receptor recruitment into nanodomains is indispensable for their function during host cell infection.

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Fliegmann, Judith, Alain Jauneau, Carole Pichereaux, et al. "LYR3, a high-affinity LCO-binding protein ofMedicago truncatula, interacts with LYK3, a key symbiotic receptor." FEBS Letters 590, no. 10 (2016): 1477–87. http://dx.doi.org/10.1002/1873-3468.12191.

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Smit, Patrick, Erik Limpens, Rene Geurts, et al. "Medicago LYK3, an Entry Receptor in Rhizobial Nodulation Factor Signaling." Plant Physiology 145, no. 1 (2007): 183–91. http://dx.doi.org/10.1104/pp.107.100495.

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Haney, Cara H., Brendan K. Riely, David M. Tricoli, Doug R. Cook, David W. Ehrhardt, and Sharon R. Long. "Symbiotic Rhizobia Bacteria Trigger a Change in Localization and Dynamics of the Medicago truncatula Receptor Kinase LYK3." Plant Cell 23, no. 7 (2011): 2774–87. http://dx.doi.org/10.1105/tpc.111.086389.

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Mbengue, Malick, Sylvie Camut, Fernanda de Carvalho-Niebel, et al. "The Medicago truncatula E3 Ubiquitin Ligase PUB1 Interacts with the LYK3 Symbiotic Receptor and Negatively Regulates Infection and Nodulation." Plant Cell 22, no. 10 (2010): 3474–88. http://dx.doi.org/10.1105/tpc.110.075861.

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Pietraszewska-Bogiel, Anna, Benoit Lefebvre, Maria A. Koini, et al. "Interaction of Medicago truncatula Lysin Motif Receptor-Like Kinases, NFP and LYK3, Produced in Nicotiana benthamiana Induces Defence-Like Responses." PLoS ONE 8, no. 6 (2013): e65055. http://dx.doi.org/10.1371/journal.pone.0065055.

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Zhukov, Vladimir, Simona Radutoiu, Lene H. Madsen, et al. "The Pea Sym37 Receptor Kinase Gene Controls Infection-Thread Initiation and Nodule Development." Molecular Plant-Microbe Interactions® 21, no. 12 (2008): 1600–1608. http://dx.doi.org/10.1094/mpmi-21-12-1600.

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Phenotypic characterization of pea symbiotic mutants has provided a detailed description of the symbiosis with Rhizobium leguminosarum bv. viciae strains. We show here that two allelic non-nodulating pea mutants, RisNod4 and K24, are affected in the PsSym37 gene, encoding a LysM receptor kinase similar to Lotus japonicus NFR1 and Medicago truncatula LYK3. Phenotypic analysis of RisNod4 and K24 suggests a role for the SYM37 in regulation of infection-thread initiation and nodule development from cortical-cell division foci. We show that RisNod4 plants carrying an L to F substitution in the LysM1 domain display a restrictive symbiotic phenotype comparable to the PsSym2A lines that distinguish ‘European’ and ‘Middle East’ Rhizobium leguminosarum bv. viciae strains. RisNod4 mutants develop nodules only in the presence of a ‘Middle East’ Rhizobium strain producing O-acetylated Nod factors indicating the SYM37 involvement in Nod-factor recognition. Along with the PsSym37, a homologous LysM receptor kinase gene, PsK1, was isolated and characterized. We show that PsK1 and PsSym37 are genetically linked to each other and to the PsSym2 locus. Allelic complementation analyses and sequencing of the extracellular regions of PsSym37 and PsK1 in several ‘European’ and ‘Afghan’ pea cultivars point towards PsK1 as possible candidate for the elusive PsSym2 gene.

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Cheval, Cécilia, Sebastian Samwald, Matthew G. Johnston, et al. "Chitin perception in plasmodesmata characterizes submembrane immune-signaling specificity in plants." Proceedings of the National Academy of Sciences 117, no. 17 (2020): 9621–29. http://dx.doi.org/10.1073/pnas.1907799117.

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The plasma membrane (PM) is composed of heterogeneous subdomains, characterized by differences in protein and lipid composition. PM receptors can be dynamically sorted into membrane domains to underpin signaling in response to extracellular stimuli. In plants, the plasmodesmal PM is a discrete microdomain that hosts specific receptors and responses. We exploited the independence of this PM domain to investigate how membrane domains can independently integrate a signal that triggers responses across the cell. Focusing on chitin signaling, we found that responses in the plasmodesmal PM require the LysM receptor kinases LYK4 and LYK5 in addition to LYM2. Chitin induces dynamic changes in the localization, association, or mobility of these receptors, but only LYM2 and LYK4 are detected in the plasmodesmal PM. We further uncovered that chitin-induced production of reactive oxygen species and callose depends on specific signaling events that lead to plasmodesmata closure. Our results demonstrate that distinct membrane domains can integrate a common signal with specific machinery that initiates discrete signaling cascades to produce a localized response.

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Stewart, Bruce, and Jonathan Bardon. "Lyke Allwaye to Contynue?" Books Ireland, no. 175 (1994): 46. http://dx.doi.org/10.2307/20626843.

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Zhou, Qinjun, Yongqiang Wei, Xiaolei Wei, Qi Wei, and Ru Feng. "CD44 over Expression Caused Drug Resistance in Activated B Cell-like Diffuse Large B-Cell Lymphoma." Blood 128, no. 22 (2016): 5316. http://dx.doi.org/10.1182/blood.v128.22.5316.5316.

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Abstract CD44, a transmembrane glycocoptotein, involved in tumor cell survival, migration , invasion, metastasis and prognosis of many cancers. This study was performed to investigate the effects of CD44 over expression on the biological function and the chemotherapeutic sensitivity of ABC-DLBCL. The full length CD44 cDNA was cloned into pEASY-T vector and then transfected into activated B cell-like diffuse large B-cell lymphoma cell line OCI-ly3. QPCR and western blot confirmed that the CD44 expression was up-regulated in both mRNA and protein levels in CD44-transfected OCI-ly3 cells. The cell proliferation of OCI-ly3-CD44 cells was significantly faster than that in OCI-ly3-GFP cells and the OCI-ly3 cells (p<0.001). Annexin V-APC/PI staining results showed that the apoptosis ratio wasn't difference among three groups (p=0.676). OCI-ly3-CD44 cells showed significantly higher migration ratio than OCI-ly3-GFP cells and the OCI-ly3 cells(p=0.031). The IC50 of doxorubicin in OCI-ly3-CD44 cells (0.348±0.072uM) was higher than that in OCI-ly3-GFP (0.348±0.072uM ) and OCI-ly3cells(0.138±0.029uM ) ( P<0.001). After treatment with doxorubicin for 24h, the OCI-ly3-CD44 cells showed lower apoptotic ratio than OCI-ly3-GFP and OCI-ly3 cells ( (17.5±1.33)% VS. (41.7±9.91) %, (41.8±7.23)%), P<0.001). In conclusion, CD44 plays a key role in cell growth regulation and chemo-resistance and is a potential target to overcome drug resistance and improve prognosis in DLBCL. Disclosures No relevant conflicts of interest to declare.

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Mosa, Rasha, Lili Huang, Hongzhuo Li, Michael Grist, Derek LeRoith, and Chen Chen. "Long-term treatment with the ghrelin receptor antagonist [d-Lys3]-GHRP-6 does not improve glucose homeostasis in nonobese diabetic MKR mice." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 314, no. 1 (2018): R71—R83. http://dx.doi.org/10.1152/ajpregu.00157.2017.

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Long-term treatment with the ghrelin receptor antagonist [d-Lys3]-GHRP-6 does not improve glucose homeostasis in nonobese diabetic MKR mice. Am J Physiol Regul Integr Comp Physiol 314: R71–R83, 2018. First published September 13, 2017; doi: 10.1152/ajpregu.00157.2017 .—Ghrelin secretion has been associated with increased caloric intake and adiposity. The expressions of ghrelin and its receptor (GHS-R1a) in the pancreas has raised the interest about the role of ghrelin in glucose homeostasis. Most of the studies showed that ghrelin promoted hyperglycemia and inhibited insulin secretion. This raised the interest in using GHS-R1a antagonists as therapeutic targets for type 2 diabetes. Available data of GHS-R antagonists are on a short-term basis. Moreover, the complexity of GHS-R1a signaling makes it difficult to understand the mechanism of action of GHS-R1a antagonists. This study examined the possible effects of long-term treatment with a GHS-R1a antagonist, [d-Lys3]-growth hormone-releasing peptide (GHRP)-6, on glucose homeostasis, food intake, and indirect calorimetric parameters in nonobese diabetic MKR mice. Our results showed that [d-Lys3]-GHRP-6 (200 nmol/mouse) reduced pulsatile growth hormone secretion and body fat mass as expected but worsened glucose and insulin intolerances and increased cumulative food intake unexpectedly. In addition, a significant increase in blood glucose and decreases in plasma insulin and C-peptide levels were observed in MKR mice following long-term [d-Lys3]-GHRP-6 treatment, suggesting a direct inhibition of insulin secretion. Immunofluorescence staining of pancreatic islets showed a proportional increase in somatostatin-positive cells and a decrease in insulin-positive cells in [d-Lys3]-GHRP-6-treated mice. Furthermore, [d-Lys3]-GHRP-6 stimulated food intake on long-term treatment via reduction of proopiomelanocortin gene expression and antagonized GH secretion via reduced growth hormone-releasing hormone gene expression in hypothalamus. These results demonstrate that [d-Lys3]-GHRP-6 is not completely opposite to ghrelin and may not be a treatment option for type 2 diabetes.

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Aghdam Shahryar, Habib, and Alireza Lotfi. "Effects of peripheral administration of ghrelin antagonist [D-Lys<sup>3</sup>]-GHRP-6 on growth performance and blood biochemical indices in broiler chickens." Archives Animal Breeding 59, no. 1 (2016): 113–19. http://dx.doi.org/10.5194/aab-59-113-2016.

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Abstract. In the present study, possible effects of peripheral administration of ghrelin antagonist [D-Lys3]-GHRP-6 on chicken performance, thyroid hormones level and serum biochemical parameters were investigated. Broiler chicks divided into five experimental groups were reared up to day 42. On day 21, a treatment was assigned to the five groups: group 1 (control), chickens without any administration of peptide or solution; group 2 (G50), chickens with intraperitoneal (IP) injection of 50 ng per 100 g body weight (BW) of D-Lys3 peptide on day 21; group 3 (G100), chickens with IP injection of 100 ng per 100 g BW of D-Lys3 peptide on day 21; group 4 (G150), chickens with IP injection of 150 ng per 100 g BW of D-Lys3 peptide on day 21; and group 5 (G200), chickens with IP injection of 200 ng per 100 g BW of D-Lys3 peptide on day 21. On days 21 (post-injection) and 42 (post-rearing), blood samples were obtained from the animals for laboratory analyses. Experimental groups administered the GHS-R antagonist showed less feed intake – i.e., administration of greater doses led to less feed intake (P < 0.01). Daily weight gains within groups G150 and G200 decreased (P < 0.01) in comparison with the control. The feed conversion ratio (FCR) did not differ among the groups. There was a significant difference between control and experimental groups for glucose, total cholesterol and phosphorus levels (P < 0.01) in post-injection samples. In post-injection and post-rearing blood samples, the thyroid hormone (T3 and T4) in 6 h increased in treated groups in comparison with the control (P < 0.01). The infusion of ghrelin antagonist [D-Lys3]-GHRP-6 reduces feed intake and body weight. With regard to increase in T4 level, it can be inferred that [D-Lys3]-GHRP-6 may increase metabolic rate, lipolysis and weight loss, which is similar to results obtained in mammalian species.

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Vera, Matthew D., Amy J. Pfizenmayer, Xiaobin Ding, Dong Xiao, and Madeleine M. Joullié. "[Lys3]didemnins as potential affinity ligands." Bioorganic & Medicinal Chemistry Letters 11, no. 1 (2001): 13–16. http://dx.doi.org/10.1016/s0960-894x(00)00585-0.

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Karpova, Inessa V., Eugenii R. Bychkov, Ilia Yu Tissen, Andrei A. Lebedev, and Petr D. Shabanov. "The effect of the ghrelin receptors inhibitor [D-Lys3]-GHRP-6 on the levels and metabolism of monoamines in symmetric brain areas of rats treated chronically with alcohol." Reviews on Clinical Pharmacology and Drug Therapy 15, no. 3 (2017): 48–56. http://dx.doi.org/10.17816/rcf15348-56.

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Aim. In the course of the study, the impact of the ghrelin receptor GHS-R1a on the condition of symmetric monoaminergic systems of the rat brain was investigated. In particular, it was intended to find out whether the treatment with the ghrelin receptor antagonist [D-Lys3]-GHRP-6, recover the original content of monoamines and their metabolites in the brain of chronic alcoholic rats. Methods. The experiments were performed on 22 Wistar male rats. Experimental animals instead of drinking water received 10 % ethanol solution. Rats of the control groups continued to consume tap water. 6 months after the beginning of forced chronic alcohol treatement, 6 rats treated with alcohol, and 6 rats received water, in a month, once in three days, were instilled intranasally with the ghrelin antagonist [D-Lys3]-GHRP-6 (1 мкг/мкл, with 10 µl to each nostril). The other animals in the same manner were administered an equivalent volume of saline. 80 minutes after the last intranasal administration of drugs, rats were decapitated. With the HPLC-method, in the hypothalamus, olfactory tubercle, striatum and hippocampus of the left and right sides of the brain the contents of noradrenaline (NA), dopamine (DA), dioxyphenylacetic acid (DOPAC), homovanillic acid (HVA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were measured. The results were processed by Student’s t-test using the statistical software package GraphPad Prism 6.0. Results. In the control rats (not exposed to either ethanol or drug) in the left striatum revealed a significant predominance of 5-HIAA compared to the same parameter of the other side of the brain. Under the condition of chronic ethanol intake, the initial left-sided asymmetry disappeared. Ethanol increased the content of 5-HT in the left hippocampus, 5-HIAA in the right olfactory tubercle and DA – in the right hypothalamus. [D-Lys3]-GHRP-6, when administered intranasally to the intact rats, significantly increased the 5-HIAA/5-HT ratio in the right olfactory tubercle, and the 5-HIAA, DOPAC and HVA levels – in the right striatum. In contrast, the left-sided effects in hippocampus were observed: the 5-HT levels increased and the 5-HIAA/5-HT ratio decreased. When instilled to intact rats, [D-Lys3]-GHRP-6 does not alter the monoaminergic systems of the hypothalamus. Between the monoaminergic systems of intact animals and alcoholic rats treated with [D-Lys3]-GHRP-6, the significant differences were shown. So, in the left hippocampus of alcoholic rats treated with [D-Lys3]-GHRP-6, the 5-HT level was higher, and the 5-HIAA/5-HT ratio was lower than in the control intact animals. Besides, in the right striatum of alcoholic rats treated with [D-Lys3]-GHRP-6, the DA metabolites levels were higher than those in the intact control animals. When comparing two groups of rats treated with [D-Lys3]-GHRP-6 (consumed water and alcoholic), the only difference was found: the alcoholic animals the content of DA in the left hypothalamus was lower than that of rats consumed water. Conclusion. Thus, by its influence on the monoaminergic system of the brain, [D-Lys3]-GHRP-6 is not an antagonist of the ethanol. Rather ethanol, when administered chronically, reduces the reactivity of the majority of monoaminergic systems to the ghrelin antagonist. Herewith, the forced chronic treatement with ethanol selectively increases the sensitivity to the [D-Lys3]-GHRP-6 in the hypothalamus DA-ergic system

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Malkov, Nikita, Judith Fliegmann, Charles Rosenberg, et al. "Molecular basis of lipo-chitooligosaccharide recognition by the lysin motif receptor-like kinase LYR3 in legumes." Biochemical Journal 473, no. 10 (2016): 1369–78. http://dx.doi.org/10.1042/bcj20160073.

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LYR3 [LysM (lysin motif) receptor-like kinase 3] of Medicago truncatula is a high-affinity binding protein for symbiotic LCO (lipo-chitooligosaccharide) signals, produced by rhizobia bacteria and arbuscular mycorrhizal fungi. The present study shows that LYR3 from several other legumes, but not from two Lupinus species which are incapable of forming the mycorrhizal symbiosis, bind LCOs with high affinity and discriminate them from COs (chitooligosaccharides). The biodiversity of these proteins and the lack of binding to the Lupinus proteins were used to identify features required for high-affinity LCO binding. Swapping experiments between each of the three LysMs of the extracellular domain of the M. truncatula and Lupinus angustifolius LYR3 proteins revealed the crucial role of the third LysM in LCO binding. Site-directed mutagenesis identified a tyrosine residue, highly conserved in all LYR3 LCO-binding proteins, which is essential for high-affinity binding. Molecular modelling suggests that it may be part of a hydrophobic tunnel able to accommodate the LCO acyl chain. The lack of conservation of these features in the binding site of plant LysM proteins binding COs provides a mechanistic explanation of how LCO recognition might differ from CO perception by structurally related LysM receptors.

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Teves, Franco, Mónica Lamas-Maceiras, Carlos García-Estrada, et al. "Transcriptional upregulation of four genes of the lysine biosynthetic pathway by homocitrate accumulation in Penicillium chrysogenum: homocitrate as a sensor of lysine-pathway distress." Microbiology 155, no. 12 (2009): 3881–92. http://dx.doi.org/10.1099/mic.0.031005-0.

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The lysine biosynthetic pathway has to supply large amounts of α-aminoadipic acid for penicillin biosynthesis in Penicillium chrysogenum. In this study, we have characterized the P. chrysogenum L2 mutant, a lysine auxotroph that shows highly increased expression of several lysine biosynthesis genes (lys1, lys2, lys3, lys7). The L2 mutant was found to be deficient in homoaconitase activity since it was complemented by the Aspergillus nidulans lysF gene. We have cloned a gene (named lys3) that complements the L2 mutation by transformation with a P. chrysogenum genomic library, constructed in an autonomous replicating plasmid. The lys3-encoded protein showed high identity to homoaconitases. In addition, we cloned the mutant lys3 allele from the L2 strain that showed a G1534 to A1534 point mutation resulting in a Gly495 to Asp495 substitution. This mutation is located in a highly conserved region adjacent to two of the three cysteine residues that act as ligands to bind the iron–sulfur cluster required for homoaconitase activity. The L2 mutant accumulates homocitrate. Deletion of the lys1 gene (homocitrate synthase) in the L2 strain prevented homocitrate accumulation and reverted expression levels of the four lysine biosynthesis genes tested to those of the parental prototrophic strain. Homocitrate accumulation seems to act as a sensor of lysine-pathway distress, triggering overexpression of four of the lysine biosynthesis genes.

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Scuto, Anna, Maciej Kujawski, Claudia Kowolik, Hua Yu, Stephen Forman, and Richard Jove. "STAT3 Silencing as a Novel Therapeutic Strategy for Activated B Cell-Type Diffuse Large B-Cell Lymphoma." Blood 114, no. 22 (2009): 926. http://dx.doi.org/10.1182/blood.v114.22.926.926.

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Abstract Abstract 926 Among the non-Hodgkin's lymphomas, the diffuse large B cell lymphoma (DLBCL) represents the most frequent (30%) of the aggressive lymphomas. Persistent STAT3 signaling contributes to malignant progression in many diverse human tumors. IL-6 and IL-10 are major activators of STAT3 signaling and are important in the pathophysiology of DLBCL. STAT3 has been found to be persistently active in activated B cells (ABC), which are non-germinal center-derived DLBCL cells. We studied the consequences of STAT3 inhibition on multiple biological functions in two representative human cell lines of this group, Ly3 and Ly10 cells. For this purpose, we established stably transduced STAT3 shRNA-expressing lentivirus Ly3 cells, control lentivirus Ly3 cells, STAT3 shRNA-expressing lentivirus Ly10 cells and control lentivirus Ly10 cells. The stable expression of STAT3 shRNA results in 40-50% reduction of total STAT3 protein levels in the STAT3 shRNA lentivirus Ly3 cells compared to the control lentivirus cells. STAT3 down-regulation induced inhibition of cell proliferation (approximately 40%). Ly3 cells respond to IL-10 more than to IL-6 in terms of proliferation; both cytokines induced less proliferation in the STAT3 shRNA lentivirus Ly3 cells compared to the control lentivirus Ly3 cells. Similar results were obtained in Ly10 cells, which respond more to IL-6 than to IL-10 in terms of proliferation. We analyzed by quantitative real-time PCR the mRNA levels of different STAT3 target genes and observed significant reduction in mRNA levels of Mcl-1, Bcl-xL and Survivin in STAT3 shRNA lentivirus Ly3 cells, as well as significant reduction of Cyclin D2 and up-regulation of STAT1 in shRNA lentivirus Ly10 cells. Comparison of these gene expression profiles with data obtained from other B-cell lymphoma cell lines revealed that silencing of STAT3 resulted in down-regulation of different STAT3 target genes in a cell-dependent manner. We also observed that both STAT3 and control lentivirus Ly3 cells have the same protein levels of c-Myc; nevertheless STAT3 silencing resulted in inhibition of IL-10-inducible upregulation of c-Myc. We next investigated the effect of STAT3 inhibition on adhesion to bone marrow stroma and chemotaxis. STAT3 shRNA lentivirus Ly3 cells adhered less to the stroma layer than control cells, and the longer they were cocultured with the stroma cells in the presence of serum-free media the more they lost the ability to adhere. Moreover, STAT3 shRNA lentivirus Ly3 cells had decreased capacity to migrate toward SDF-1 alpha, an important factor that mediates proliferation, survival, chemotaxis, migration and adhesion into bone marrow stroma. Radiation, in combination with chemotherapy, is one of the therapies used for DLBCL patients. We therefore investigated whether STAT3 down-regulation sensitized Ly3 cells to radiation. Radiation induced a higher accumulation of phospho-H2A.X (first sentinel event following DNA damage such as DSBs) and apoptosis in STAT3 shRNA lentivirus cells compared to control cells. Moreover, IL-6 and IL-10 protected the STAT3 shRNA lentivirus Ly3 cells less than the control cells from the induction of phospho-H2A.X following radiation. We further investigated the effect of STAT3 silencing in animal models of Ly3 lymphoma (Nude or NOD-SCID mice). Tumors in control lentivirus Ly3-bearing mice grew robustly, whereas tumors in STAT3 shRNA lentivirus Ly3-bearing mice regressed 5 days after injection. This tumor regression was associated with Caspase-3-dependent apoptosis, significant reduction of STAT3 target genes at the protein level such as Mcl-1, c-Myc and Survivin (approximately 40% to 60% inhibition), and reduction of cytokine production such as IL-10, IL-15, Leptin and Thrombopoietin. Taken together, these results suggest that inhibition of STAT3 is a potential promising approach in the therapy of ABC-type DLBCL. Disclosures: No relevant conflicts of interest to declare.

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Kamau, Edwin, Nick D. Tsihlis, L. Alice Simmons, and Anne Grove. "Surface salt bridges modulate the DNA site size of bacterial histone-like HU proteins." Biochemical Journal 390, no. 1 (2005): 49–55. http://dx.doi.org/10.1042/bj20050274.

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Bacterial histone-like DNA-binding proteins are best known for their role in compacting the genomic DNA. Of these proteins, HU is ubiquitous and highly conserved across the eubacterial kingdom. Using the HBsu (Bacillus subtilis-encoded HU homologue) as a model, we explore here the molecular basis for the ability of some HU homologues to engage a longer approx. 35 bp DNA site as opposed to the much shorter sites reported for other homologues. Using electrophoretic mobility-shift assays, we show that the DNA site size for HBsu is approx. 10–13 bp and that a specific surface salt bridge limits the DNA site size for HBsu. Surface exposure of the highly conserved Lys3, achieved by substitution of its salt-bridging partner Asp26 with Ala, leads to enhanced DNA compaction by HBsu-D26A (where D26A stands for the mutant Asp26→Ala), consistent with the interaction of Lys3 with the ends of a 25 bp duplex. Both HBsu and HBsu-D26A bend DNA, as demonstrated by their equivalent ability to promote ligase-mediated DNA cyclization, indicating that residues involved in mediating DNA kinks are unaltered in the mutant protein. We suggest that Lys3 is important for DNA wrapping due to its position at a distance from the DNA kinks where it can exert optimal leverage on flanking DNA and that participation of Lys3 in a surface salt bridge competes for its interaction with DNA phosphates, thereby reducing the occluded site size.

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Degeest, Bart, and Luc De Vuyst. "Indication that the Nitrogen Source Influences Both Amount and Size of Exopolysaccharides Produced by Streptococcus thermophilus LY03 and Modelling of the Bacterial Growth and Exopolysaccharide Production in a Complex Medium." Applied and Environmental Microbiology 65, no. 7 (1999): 2863–70. http://dx.doi.org/10.1128/aem.65.7.2863-2870.1999.

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ABSTRACT Streptococcus thermophilus LY03 is a yogurt strain producing the same exopolysaccharide material in both milk and MRS broth. Actually, two types of exopolysaccharides are produced simultaneously. The two exopolysaccharides are identical in monomer composition (galactose and glucose in a 4:1 ratio) but differ in molecular size. Gel permeation chromatography revealed a high-molecular-mass exopolysaccharide (1.8 × 106) and a low-molecular-mass exopolysaccharide (4.1 × 105). Both exopolysaccharides can be isolated from the fermentation broth separately. The proportion in which they are produced is strongly dependent on the carbon/nitrogen ratio of the fermentation broth. A shift from a high-molecular-mass exopolysaccharide to a low-molecular-mass exopolysaccharide was observed with increasing initial complex nitrogen concentrations. All necessary biokinetic parameters to study the kinetics of S. thermophilus LY03 fermentations were obtained from a mathematical model which describes both S. thermophilus LY03 growth and exopolysaccharide production and degradation. The model is valid with various initial complex nitrogen concentrations and can be applied to simulate exopolysaccharide production in a milk medium.

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Luan, Yepeng, Chunhua Ma, Zhongguo Sui, et al. "LYP3, a New Bestatin Derivative for Aminopeptidase N Inhibition." Medicinal Chemistry 7, no. 1 (2011): 32–36. http://dx.doi.org/10.2174/157340611794072706.

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Kuroda, Kayuri, Huang Hequing, Anupom Mondal, et al. "Ghrelin Is an Essential Factor for Motilin-Induced Gastric Contraction in Suncus murinus." Endocrinology 156, no. 12 (2015): 4437–47. http://dx.doi.org/10.1210/en.2015-1561.

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Motilin was discovered in the 1970s as the most important hormone for stimulating strong gastric contractions; however, the mechanisms by which motilin causes gastric contraction are not clearly understood. Here, we determined the coordinated action of motilin and ghrelin on gastric motility during fasted and postprandial contractions by using house musk shrew (Suncus murinus; order: Insectivora, suncus named as the laboratory strain). Motilin-induced gastric contractions at phases I and II of the migrating motor complex were inhibited by pretreatment with (d-Lys3)-GHRP-6 (6 mg/kg/h), a ghrelin receptor antagonist. Administration of the motilin receptor antagonist MA-2029 (0.1 mg/kg) and/or (d-Lys3)-GHRP-6 (0.6 mg/kg) at the peak of phase III abolished the spontaneous gastric phase III contractions in vivo. Motilin did not stimulate gastric contractions in the postprandial state. However, in the presence of a low dose of ghrelin, motilin evoked phase III–like gastric contractions even in the postprandial state, and postprandial gastric emptying was accelerated. In addition, pretreatment with (d-Lys3)-GHRP-6 blocked the motilin-induced gastric contraction in vitro and in vivo, and a γ-aminobutyric acid (GABA) antagonist reversed this block in gastric contraction. These results indicate that blockade of the GABAergic pathway by ghrelin is essential for motilin-induced gastric contraction.

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Sulima, Anton S., Vladimir A. Zhukov, Olga A. Kulaeva, Ekaterina N. Vasileva, Alexey Y. Borisov, and Igor A. Tikhonovich. "New sources of Sym2A allele in the pea (Pisum sativum L.) carry the unique variant of candidate LysM-RLK gene LykX." PeerJ 7 (November 20, 2019): e8070. http://dx.doi.org/10.7717/peerj.8070.

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At the onset of legume-rhizobial symbiosis, the mutual recognition of partners occurs based on a complicated interaction between signal molecules and receptors. Bacterial signal molecules named Nod factors (“nodulation factors”) are perceived by the plant LysM-containing receptor-like kinases (LysM-RLKs) that recognize details of its structure (i.e., unique substitutions), thus providing the conditions particular to symbiosis. In the garden pea (Pisum sativum L.), the allelic state of Sym2 gene has long been reported to regulate the symbiotic specificity: for infection to be successful, plants with the Sym2A allele (for “Sym2 Afghan”, as these genotypes originate mostly from Afghanistan) require an additional acetylation of the Nod factor which is irrelevant for genotypes with the Sym2E allele (for “Sym2 European”). Despite being described about 90 years ago, Sym2 has not yet been cloned, though phenotypic analysis suggests it probably encodes a receptor for the Nod factor. Recently, we described a novel pea gene LykX (PsLykX) from the LysM-RLK gene family that demonstrates a perfect correlation between its allelic state and the symbiotic specificity of the Sym2A-type. Here we report on a series of Middle-Eastern pea genotypes exhibiting the phenotype of narrow symbiotic specificity discovered in the VIR plant genetic resources gene bank (Saint-Petersburg, Russia). These genotypes are new sources of Sym2A, as has been confirmed by an allelism test with Sym2A pea cv. Afghanistan. Within these genotypes, LykX is present either in the allelic state characteristic for cv. Afghanistan, or in another, minor allelic state found in two genotypes from Tajikistan and Turkmenistan. Plants carrying the second allele demonstrate the same block of rhizobial infection as cv. Afghanistan when inoculated with an incompatible strain. Intriguingly, this “Tajik” allele of LykX differs from the “European” one by a single nucleotide polymorphism leading to an R75P change in the receptor part of the putative protein. Thus, our new data are in agreement with the hypothesis concerning the identity of LykX and the elusive Sym2 gene.

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Abdel-Wahab, Yasser H. A., Gavin J. Power, Ming T. Ng, Peter R. Flatt, and J. Michael Conlon. "Insulin-releasing properties of the frog skin peptide pseudin-2 and its [Lys18]-substituted analogue." Biological Chemistry 389, no. 2 (2008): 143–48. http://dx.doi.org/10.1515/bc.2008.018.

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Abstract Pseudin-2 is a cationic α-helical peptide that was first isolated from the skin of the paradoxical frog Pseudis paradoxa on the basis of its antimicrobial activity. We have investigated the insulin-releasing properties and cytotoxicity of the peptide, together with selected analogues with increased cationicity and hydrophobicity. At concentrations in the range 10-9–10-6 m, pseudin-2, and its [Lys18], [Phe8], and [d-Lys3,d-Lys10,d-Lys14] derivatives, stimulated insulin release from the BRIN-BD11 clonal β-cell line without increasing release of lactate dehydrogenase. The [Lys18] analogue was the most potent (46% increase in insulin release at 10-9 m) and the most effective (215% increase in insulin release at 10-6 m). The more cationic [Lys3,Lys10,Lys14] and [Lys3,Lys10,Lys14,Lys21] analogues lacked insulinotropic action and the more hydrophobic [Phe16] analogue was cytotoxic at concentrations ≥10-7 m. Pseudin-2 and [Lys18]-pseudin-2 had no effect on intracellular calcium concentrations and stimulated insulin release in the absence of external calcium. [Lys18]-pseudin-2 (10-8 m) stimulated insulin release in the presence of diazoxide and verapamil. Our results demonstrate that pseudin-2 stimulates insulin secretion from BRIN-BD11 cells by a mechanism involving Ca2+-independent pathways and identify [Lys18]-pseudin-2 as a peptide that may have potential for development as a therapeutically valuable insulinotropic agent for the treatment of type 2 diabetes.

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Antonenko, Yuri N., Grigory S. Gluhov, Alexander M. Firsov, et al. "Gramicidin A disassembles large conductive clusters of its lysine-substituted derivatives in lipid membranes." Physical Chemistry Chemical Physics 17, no. 26 (2015): 17461–70. http://dx.doi.org/10.1039/c5cp02047f.

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Gramicidin A (gA) blocks dye leakage from liposomes induced by [Lys3]gA, thereby highlighting the importance of cation–π interactions for pore formation. Based on cryo-em, large pores are attributed to 40 Å-diameter peptide clusters.

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Ferro-Flores, G., I. A. Rivero, C. L. Santos-Cuevas, et al. "Click chemistry for [99mTc(CO)3] labeling of Lys3-bombesin." Applied Radiation and Isotopes 68, no. 12 (2010): 2274–78. http://dx.doi.org/10.1016/j.apradiso.2010.06.014.

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Clarke, ND, LJ Beamer, HR Goldberg, C. Berkower, and CO Pabo. "The DNA binding arm of lambda repressor: critical contacts from a flexible region." Science 254, no. 5029 (1991): 267–70. http://dx.doi.org/10.1126/science.254.5029.267.

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Segments of protein that do not adopt a well-ordered conformation in the absence of DNA can still contribute to site-specific recognition of DNA. The first six residues (NH2-Ser1-Thr2-Lys3-Lys4-Lys5-Pro6-) of phage lambda repressor are flexible but are important for site-specific binding. Low-temperature x-ray crystallography and codondirected saturation mutagenesis were used to study the role of this segment. All of the functional sequences have the form [X]1-[X]2-[Lys or Arg]3-[Lys]4-[Lys or Arg]5-[X]6. A high-resolution (1.8 angstrom) crystal structure shows that Lys3 and Lys4 each make multiple hydrogen bonds with guanines and that Lys5 interacts with the phosphate backbone. The symmetry of the complex breaks down near the center of the site, and these results suggest a revision in the traditional alignment of the six lambda operator sites.

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Лебедев, А. А., П. П. Хохлов, Н. Д. Якушина та ін. "Фармакологический и биохимический анализ участия пептидной системы грелина в поведенческих проявлениях игровой зависимости у крыс". Экспериментальная и клиническая фармакология 82, № 6 (2019): 16–20. http://dx.doi.org/10.30906/0869-2092-2019-82-6-16-20.

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Целью исследования было изучение влияния активации и блокады рецепторов грелина на проявление импульсивного компонента игровой зависимости и содержания эндогенного дезацил-грелина (ДАГ) в лимбических структурах головного мозга у крыс. Рабочей гипотезой было допущение, что грелин участвует в формировании аддиктивных форм поведения и злоупотребления веществами на фоне стрессорных факторов или сигналов внешней среды. Крыс в течение 3 недель обучали побежкам в 3-лучевом лабиринте. В рукаве 1 каждая побежка подкреплялась 1 семенем подсолнечника, в рукаве 2 — каждая вторая побежка подкреплялась 2 семенами, в рукаве 3 — каждая третья побежка в рукаве 3 подкреплялась 3 семенами. В качестве фармакологических веществ использовали грелин и его антагонист [D-Lys3]-GHRP-6, которые вводили интраназально в дозе 10 мкг в 20 мкл. После 7-дневного введения пептидных препаратов крыс декапитировали и выделяли структуры мозга (миндалина, гиппокамп и гипоталамус). С помощью высокочувствительного иммуноферментного анализа оценивали содержание ДАГ. В поведенческих опытах показано, что введение грелина повышает число побежек в рукав с низкой вероятностью, но большей величиной подкрепления, в то время как [D-Lys3]-GHRP-6 увеличивает число побежек в рукав с высокой (100 %) вероятностью, но меньшей величиной подкрепления. Концентрация ДАГ в структурах мозга контрольных крыс варьировала: в миндалине она была в 1,5 раза, а в гипоталамусе — в 3 раза выше, чем в гиппокампе. После курса введения грелина содержание ДАГ повышалось в гиппокампе в 1,5 раза и снижалось в гипоталамусе в 2 раза. D-Lys3-GHRP-6 значимо (в 3 раза) снижал содержание ДАГ только в гипоталамусе. Сделан вывод о том, что система пептидов грелина участвует в игровом поведении, влияя в основном на импульсивный компонент игровой зависимости.

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O'Connor, Brian P., and Edouard S. St. Pierre. "Older Persons' Perceptions of the Frequency and Meaning of Elderspeak from Family, Friends, and Service Workers." International Journal of Aging and Human Development 58, no. 3 (2004): 197–221. http://dx.doi.org/10.2190/ly83-kpxd-h2f2-jrq5.

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Zich, Jane M., C. Clifford Attkisson, and Thomas K. Greenfield. "Screening for Depression in Primary Care Clinics: The CES-D and the BDI." International Journal of Psychiatry in Medicine 20, no. 3 (1990): 259–77. http://dx.doi.org/10.2190/lykr-7vhp-yjem-mkm2.

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Wallerstein, Robert, and Jane Milton. "Plenary session: ‘The link and the other’." International Journal of Psychoanalysis 83, no. 3 (2002): 682–84. http://dx.doi.org/10.1516/ly33-kflb-n132-4yum.

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BALLAS, APOSTOLOS, and VASILIS THEOHARAKIS. "Exploring Diversity in Accounting through Faculty Journal Perceptions." Contemporary Accounting Research 20, no. 4 (2003): 619–44. http://dx.doi.org/10.1506/mlwh-kbtm-et47-lykh.

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Lund, Anders H., Jörg Schmidt, Arne Luz, Annette Balle Sørensen, Mogens Duch, and Finn Skou Pedersen. "Replication and Pathogenicity of Primer Binding Site Mutants of SL3-3 Murine Leukemia Viruses." Journal of Virology 73, no. 7 (1999): 6117–22. http://dx.doi.org/10.1128/jvi.73.7.6117-6122.1999.

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ABSTRACT Retroviral reverse transcription is primed by a cellular tRNA molecule annealed to an 18-bp primer binding site sequence. The sequence of the primer binding site coincides with that of a negatively acting cis element that mediates transcriptional silencing of murine leukemia virus (MLV) in undifferentiated embryonic cells. In this study we test whether SL3-3 MLV can replicate stably using tRNA primers other than the cognate tRNAPro and analyze the effect of altering the primer binding site sequence to match the 3′ end of tRNA1 Gln, tRNA3 Lys, or tRNA1,2 Arg in a mouse pathogenicity model. Contrary to findings from cell culture studies of primer binding site-modified human immunodeficiency virus type 1 and avian retroviruses, our findings were that SL3-3 MLV may stably and efficiently replicate with tRNA primers other than tRNAPro. Although lymphoma induction of the SL3-3 Lys3 mutant was significantly delayed relative to that of the wild-type virus, molecular tumor analysis indicated that all the primer binding site-modified viruses induce T-cell lymphomas similar to those induced by the wild-type virus in terms of frequencies of genomic rearrangements within the T-cell receptor β-chain, the immunoglobulin κ light chain, and the c-myc locus. Whereas none of the mutants were found to revert to tRNAProprimer utilization, in two tumors resulting from the injection of the SL3-3 Lys3 mutant the primer binding site was altered to match that of a new primer species, tRNA1,2 Lys. In addition, recombination with endogenous viruses resulting in the generation of recombinant viruses carrying a glutamine primer binding site was detected in the majority of the tumors induced by the SL3-3 Lys3 mutant as well as in two tumors induced by wild-type SL3-3 and the SL3-3 Arg1,2 mutant.

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Pei, Xiao M., Benjamin Y. Yung, Shea Ping Yip, Michael Ying, Iris F. Benzie, and Parco M. Siu. "Desacyl ghrelin prevents doxorubicin-induced myocardial fibrosis and apoptosis via the GHSR-independent pathway." American Journal of Physiology-Endocrinology and Metabolism 306, no. 3 (2014): E311—E323. http://dx.doi.org/10.1152/ajpendo.00123.2013.

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Doxorubicin is an effective chemotherapeutic agent used to treat malignancies, but it causes cardiomyopathy. Preliminary evidence suggests that desacyl ghrelin might have protective effects on doxorubicin cardiotoxicity. This study examined the cellular effects of desacyl ghrelin on myocardial fibrosis and apoptosis in a doxorubicin cardiomyopathy experimental model. Adult C57BL/6 mice received an intraperitoneal injection of doxorubicin to induce cardiomyopathy, followed by 4-day treatment of saline (control) or desacyl ghrelin with or without [d-Lys3]-GHRP-6 (a growth hormone secretagogue receptor or GHSR1a antagonist). Ventricular structural and functional parameters were evaluated by transthoracic echocardiography. Molecular and cellular measurements were performed in ventricular muscle to examine myocardial fibrosis and apoptosis. Cardiac dysfunction was induced by doxorubicin, as indicated by significant decreases in ventricular fractional shortening and ejection fraction. This doxorubicin-induced cardiac dysfunction was prevented by the treatment of desacyl ghrelin no matter with or without the presence of [d-Lys3]-GHRP-6. Doxorubicin induced fibrosis (accumulated collagen deposition and increased CTGF), activated apoptosis (increased TUNEL index, apoptotic DNA fragmentation, and caspase-3 activity and decreased Bcl-2/Bax ratio), and suppressed phosphorylation status of prosurvival signals (ERK1/2 and Akt) in ventricular muscles. All these molecular and cellular alterations induced by doxorubicin were not found in the animals treated with desacyl ghrelin. Notably, the changes in the major markers of apoptosis, fibrosis, and Akt phosphorylation were found to be similar in the animals following the treatment of desacyl ghrelin with and without GHSR antagonist [d-Lys3]-GHRP-6. These findings demonstrate clearly that desacyl ghrelin protects the cardiomyocytes against the doxorubicin-induced cardiomyopathy by preventing the activation of cardiac fibrosis and apoptosis, and the effects are probably mediated through GHSR-independent mechanism.

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Delhanty, P. J. D., P. M. van Koetsveld, C. Gauna, et al. "Ghrelin and its unacylated isoform stimulate the growth of adrenocortical tumor cells via an anti-apoptotic pathway." American Journal of Physiology-Endocrinology and Metabolism 293, no. 1 (2007): E302—E309. http://dx.doi.org/10.1152/ajpendo.00377.2006.

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Ghrelin is expressed in normal human adrenocortical cells and induces their proliferation through growth hormone secretagogue receptor 1a (GHS-R1a). Consequently, it was of interest to us to determine whether acylated ghrelin and its predominant serum isoform, unacylated ghrelin, also act as factors for adrenocortical carcinoma cell growth. To examine a potential ghrelin-regulated system in adrenocortical tumors, we measured proliferative effects of acylated and unacylated ghrelin in the adrenocortical carcinoma cell lines SW-13 and NCI-H295R. We also examined the expression of ghrelin, GHS-R1a, and corticotrophin-releasing factor receptor 2 (CRF-R2). Acylated and unacylated ghrelin in the nanomolar range dose-dependently induced adrenocortical cell growth up to 200% of untreated controls, as measured by thymidine uptake and WST1 assay. The proliferative effects of acylated and unacylated ghrelin in SW-13 cells was blocked by [d-Lys3]growth hormone-releasing peptide 6 (GHRP6), but a CRF-R2 antagonist had no effect on unacylated ghrelin growth stimulation. Cell cycle analysis suggests that acylated and unacylated ghrelin suppress the sub-G0/apoptotic fraction by up to 50%. Measurement of DNA fragmentation and caspase-3 and -7 activity in SW-13 cells confirmed that acylated and unacylated ghrelin suppress apoptotic rate. SW-13 cells express preproghrelin mRNA and secrete ghrelin, and [d-Lys3]GHRP6 suppresses their basal proliferation rate, strongly suggesting that ghrelin could act as an auto/paracrine growth factor. Acylated and unacylated ghrelin are potential auto/paracrine factors acting through an antiapoptotic pathway to stimulate adrenocortical tumor cell growth. Unacylated ghrelin-stimulated growth is suppressed by an antagonist of GHS-R1a, suggesting either that unacylated ghrelin is acylated before its action or that ghrelin, unacylated ghrelin, and [d-Lys3]GHRP-6 bind to a novel receptor in these cells.

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Yu, Angus P., Xiao M. Pei, Thomas K. Sin, et al. "[D-Lys3]-GHRP-6 exhibits pro-autophagic effects on skeletal muscle." Molecular and Cellular Endocrinology 401 (February 2015): 155–64. http://dx.doi.org/10.1016/j.mce.2014.09.031.

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Szabó, Renáta, Rudolf Ménesi, Andor H. Molnár, et al. "New Metabolic Influencer on Oxytocin Release: The Ghrelin." Molecules 24, no. 4 (2019): 735. http://dx.doi.org/10.3390/molecules24040735.

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Background: The hypothalamic–pituitary axis by secreting neuropeptides plays a key role in metabolic homeostasis. In light of the metabolic regulation, oxytocin is a potential neuropeptide for therapies against obesity and related disorders. The aim of our study is to measure ghrelin-induced oxytocin secretion in rats and to detect the changes after administration of ghrelin antagonist. Methods: Ghrelin was administrated centrally (intracerebroventricular, i.c.v., 1.0, 10.0, and 100.0 pmol) or systemically (intravenous, i.v., 1.0, and 10.0 nmol). [d-Lys3]-GHRP-6 ghrelin antagonist was injected 15 min before ghrelin injection in a dose of 10.0 pmol i.c.v. and 10.0 nmol i.v. Results: Either i.c.v. or i.v. administration of ghrelin dose-dependently increased the plasma oxytocin concentration. Following pretreatment with the ghrelin antagonist [d-Lys3]-GHRP-6, the high plasma oxytocin level induced by ghrelin was significantly reduced. Conclusion: The results indicate that the release of oxytocin is influenced directly by the ghrelin system. Examination of the mechanism of ghrelin-induced oxytocin secretion is a new horizon for potential therapeutic options.

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Brulé, Daphnée, Clizia Villano, Laura J. Davies, et al. "The grapevine ( Vitis vinifera ) LysM receptor kinases Vv LYK 1‐1 and Vv LYK 1‐2 mediate chitooligosaccharide‐triggered immunity." Plant Biotechnology Journal 17, no. 4 (2018): 812–25. http://dx.doi.org/10.1111/pbi.13017.

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Gupta, Mamta, Guangzhen Hu, Linda E. Wellik, et al. "SHP1 Suppression In Diffuse Large B Cell Lymphoma Is Regulated Through Promoter Hypermethylation At Novel CpG2 Island and H3K27 Trimethylation Histone Mark." Blood 122, no. 21 (2013): 632. http://dx.doi.org/10.1182/blood.v122.21.632.632.

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Abstract Epigenetic silencing of tumor-suppressor genes has potential therapeutic relevance if the diagnostic and prognostic utility of particular DNA methylation patterns could be established. SHP1 is a tumor-suppressor gene that can be downregulated in various neoplasms. The aims of this study are (i) whether SHP1 was lost/ suppressed in diffuse large B cell lymphoma (DLBCL) (ii) Understanding the role of various epigenetic mechanisms including CpG hypermethylation and histone modifications in SHP1 loss, and (iii) whether SHP1 in DLBCL can be re-expressed with epigenetic therapy. We initially analyzed SHP1 mRNA expression in DLBCL (n=9) patient specimens and normal B-cells by QRT-PCR. Decreased expression of SHP1 mRNA was observed in all the DLBCL patient samples as compared to normal B cells. DLBCL tumor samples (n=37) along with normal tonsils (n=10) were stained for SHP1 protein by immunohistochemistry (IHC). SHP1 expression was lost or suppressed in 53% of DLBCL tumors compared to normal controls. These data, when taken together, confirm that SHP1 is strongly expressed in normal B-cells and can be lost or suppressed in DLBCL tumors. We performed epigenetic studies of SHP1 CpG promoter 2 (named CPG1 P2) in 37 DLBCL tumors and 6 DLBCL cell lines by methylation specific (MSP1) PCR and pyrosequencing methods. Surprisingly, SHP1 CpG1 P2 hypermethylation (0/37) was not detected in patient samples or cell lines by both methods. Our data conclusively demonstrate that SHP1 CpG1 P2 hypermethylation is absent in DLBCL tumors. The finding that SHP1 was lost or silenced in many of the clinical samples without finding any CpG1 P2 hypermethylation led us to extend our methylation studies to a novel, more proximal CpG island within P2 (named CpG2) that has not been previously studied in any hematological malignancies. We designed the primers specific for these CpG2 sites and ran the pyrosequencing on the same 37 DLBCL tumors as used above for CpG1 methylation. Using a cut off of >25% (intermediate to high methylation) 57% (21/37) of patient samples were methylated. The Ly3, Ly10, and DHL2 DLBCL cell lines were also hypermethylated at CpG2. To confirm the role of CpG2 methylation in SHP1 expression, we treated the Ly3 and DHL2 cells with the DNA methyltransferase inhibitor, 5-Azacytidine (Aza). We observed increased SHP1 expression compared to untreated control cells in Ly3 and DHL2 cells followed by a progressive demethylation of SHP1 CpG2, as shown by positive U-MSP with increasing amplification intensity. This resulted in a corresponding down-regulation of activated STAT3 in a similar time dependent manner. Treatment of Ly3 and DHL2 cells with 5-Aza for 0-6 days significantly (p=0.0008 for Ly3 and p=0.03 for DHL2) reduced the cell survival (40% decrease at day 6). To further investigate epigenetic control of SHP1 in DLBCL, we proceeded to study whether histone modifications in the SHP1 promoter were involved in the regulation of SHP1 expression. Chromatin immunoprecipitation assays were performed in the Ly3, Raji cell line and normal B cells, by using antibodies to repressing marks on histones. We observed that the SHP1 promoter in the Ly3 and Raji cells was enriched for both H3K9me3 and H3K27me3 repressing marks as compared to normal B cells. To confirm the role of histone modifications in SHP1 expression, we treated Ly3 cells with the histone methyltransferase inhibitor (HMT) DZNEP and the HDAC inhibitor LBH589 (Novartis Pharmaceuticals). DZNEP has been shown to be very specific for histone suppressive marks H3K27me3 that are mediated through EZH2. Treatment with DZNEP partially reverted the H3K27me3 histone marks in the SHP1 promoter followed by increased the SHP1 expression in Ly3 cells. LBH589 treatment was not able to inhibit H3K27me3 marks, but was able to inhibit the CpG2 methylation within the SHP1 promoter 2 alone or in combination with 5- Aza. In conclusion, these data provide a comprehensive characterization of the epigenetic mechanisms leading to SHP1 deregulation in the DLBCL tumors. These findings support the notion that control of SHP1 P2 in DLBCL tumors is associated with both DNA promoter methylation and histone methylation, which explains our results that treatment with 5-Aza, DZNEP and LBH589 could reactivate SHP1 expression and inhibit DLBCL cell survival. Therefore, SHP1 may prove useful diagnostic and prognostic marker and provide a rationale to use epigenetic therapy for patients with DLBCL. Disclosures: No relevant conflicts of interest to declare.

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Takahashi, Hideyuki, Yohei Kurose, Muneyuki Sakaida, et al. "Ghrelin differentially modulates glucose-induced insulin secretion according to feeding status in sheep." Journal of Endocrinology 194, no. 3 (2007): 621–25. http://dx.doi.org/10.1677/joe-07-0206.

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The present study was conducted to investigate roles of ghrelin in glucose-induced insulin secretion in fasting- and meal-fed state in sheep. Castrated Suffolk rams were fed a maintenance diet of alfalfa hay cubes once a day. Hyperglycemic clamp (HGC) was carried out to examine glucose-induced insulin response from 48 to 53 h (fasting state) and from 3 to 8 h (meal-fed state) after feeding in Experiment 1 and 2 respectively. Total dose of 70 nmol/kg body weight of d-Lys3-GHRP6, a GH secretagogue receptor 1a (GHS-R1a) antagonist, was intravenously administered at 0, 60, and 120 min after the commencement of HGC. In the fasting state, the ghrelin antagonist significantly (P < 0.01) enhanced glucose-induced insulin secretion. In the meal-fed state, i.v. administration of synthetic ovine ghrelin (0.04 μ g/kg body weight per min during HGC) significantly (P < 0.05) enhanced glucose-induced insulin secretion. d-Lys3-GHRP6 treatment suppressed ghrelin-induced enhancement of the insulin secretion. In conclusion, ghrelin has an inhibitory and stimulatory role in glucose-induced insulin secretion via GHS-R1a in fasting- and meal-fed state respectively.

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Shabanov, Petr D., Aleksandra A. Blazhenko, Aleksandr S. Devyashin, Platon P. Khokhlov, and Andrei A. Lebedev. "In search of new brain biomarkers of stress." Research Results in Pharmacology 7, no. 1 (2021): 41–46. http://dx.doi.org/10.3897/rrpharmacology.7.63326.

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The aim: of the study was to investigate the level of ghrelin in various brain structures during a stress response in Zebrafish to a predator, to evaluate this indicator as a potential biomarker of stress, and the effect of a benzodiazepine tranquilizer (phenazepam) on stress-induced changes Materials and methods: The object of the study was Zebrafish, or Danio rerio wild type, which was subjected to stress by exposure to a predator Hypsophrys nicaraguensis from the cichlid family. In the tail tissue, the level of cortisol was determined, in the brain – the level of total (acylated and non-acylated) ghrelin by the method of enzyme-linked immunosorbent assay. The benzodiazepine anxiolytic phenazepam (1 mg/L), a ghrelin antagonist [D-Lys3]-GHRP-6 (0.333 mg/l) and corticotropin-releasing hormone (CRF; 0.4 mg/L) were used as the pharmacological agents. Results and discussion: Exposure to a predator, just as administering CRF, more than doubled the level of cortisol in the tail tissue. [D-Lys3]-GHRP-6 and phenazepam prevented an increase in a tissue cortisol level. Simultaneously, in the medulla oblongata and cerebellum, the phylogenetically most ancient structures, rather than in the forebrain (telencephalon) or in the midbrain (corpora bigemia), the level of ghrelin was recorded about 500 pg/g of total protein. In response to exposure to a predator, the level of ghrelin increased in the forebrain and midbrain to nanogram concentrations and moderately decreased in the cerebellum. The effect was prevented by phenazepam and [D-Lys3]-GHRP-6. Conclusion: Increases in ghrelin in the brain in response to stressful situations can be seen as a functional brain biomarker of stress, along with increased levels of tissue cortisol levels. Both of these effects are prevented by both the ghrelin antagonist and the benzodiazepine tranquilizer. The mechanism of action of the tranquilizer is a functional antagonism between the GABAergic system of the brain and the ghrelin system.

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Deleon, Heriberto, Juan Garcia, Dionn Carlo Silva, Oscar Quintanilla, Zen Faulkes, and John M. Thomas. "Culturing embryonic cells from the parthenogenetic clonal marble crayfish (Marmorkrebs) Procambarus virginalis Lyko, 2017 (Decapoda: Astacidea: Cambaridae)." Journal of Crustacean Biology 39, no. 6 (2019): 758–63. http://dx.doi.org/10.1093/jcbiol/ruz063.

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Abstract The parthenogenetic marbled crayfish, or Marmorkrebs (Procambarus virginalis Lyko 2017), is an emerging model organism. We describe a method to isolate cells from early-stage embryos and culture them in vitro. The identity of the cells was confirmed by sequencing the cytochrome c oxidase subunit I (COI) gene. This technique can be applied for use in the manipulation of embryonic parthenogenetic crayfish cells.

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Maeda, Hideo, Hiroyo Nakamura, Shozo Kobori, et al. "Molecular Cloning of a Human Apolipoprotein E Variant: E5 (Glu3→Lys3)1." Journal of Biochemistry 105, no. 4 (1989): 491–93. http://dx.doi.org/10.1093/oxfordjournals.jbchem.a122692.

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Schoepp, Darryle D., James A. Monn, Gerard J. Marek, George A. Ghajanian, and Bita Moghaddam. "LY3 54740: A Systemically Active mGlu2/mGlu3 Receptor Agonist." CNS Drug Reviews 5, no. 1 (2006): 1–12. http://dx.doi.org/10.1111/j.1527-3458.1999.tb00082.x.

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Degeest, Bart, Frederik Vaningelgem, Andrew P. Laws, and Luc De Vuyst. "UDP-N-Acetylglucosamine 4-Epimerase Activity Indicates the Presence of N-Acetylgalactosamine in Exopolysaccharides of Streptococcus thermophilus Strains." Applied and Environmental Microbiology 67, no. 9 (2001): 3976–84. http://dx.doi.org/10.1128/aem.67.9.3976-3984.2001.

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ABSTRACT The monomer composition of the exopolysaccharides (EPS) produced byStreptococcus thermophilus LY03 and S. thermophilus Sfi20 were evaluated by high-pressure liquid chromatography with amperometric detection and nuclear magnetic resonance spectroscopy. Both strains produced the same EPS composed of galactose, glucose, and N-acetylgalactosamine. Further, it was demonstrated that the activity of the precursor-producing enzyme UDP-N-acetylglucosamine 4-epimerase, converting UDP-N-acetylglucosamine into UDP-N-acetylgalactosamine, is responsible for the presence of N-acetylgalactosamine in the EPS repeating units of both strains. The activity of UDP-N-acetylglucosamine 4-epimerase was higher in bothS. thermophilus strains than in a non-EPS-producing control strain. However, the level of this activity was not correlated with EPS yields, a result independent of the carbohydrate source applied in the fermentation process. On the other hand, both the amounts of EPS and the carbohydrate consumption rates were influenced by the type of carbohydrate source used during S. thermophilus Sfi20 fermentations. A correlation between activities of the enzymes α-phosphoglucomutase, UDP-glucose pyrophosphorylase, and UDP-galactose 4-epimerase and EPS yields was seen. These experiments confirm earlier observed results for S. thermophilus LY03, although S. thermophilusSfi20 preferentially consumed glucose for EPS production instead of lactose in contrast to the former strain.

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Gauna, Carlotta, Rosalie M. Kiewiet, Joop A. M. J. L. Janssen, et al. "Unacylated ghrelin acts as a potent insulin secretagogue in glucose-stimulated conditions." American Journal of Physiology-Endocrinology and Metabolism 293, no. 3 (2007): E697—E704. http://dx.doi.org/10.1152/ajpendo.00219.2007.

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Acylated and unacylated ghrelin (AG and UAG) are gut hormones that exert pleiotropic actions, including regulation of insulin secretion and glucose metabolism. In this study, we investigated whether AG and UAG differentially regulate portal and systemic insulin levels after a glucose load. We studied the effects of the administration of AG (30 nmol/kg), UAG (3 and 30 nmol/kg), the ghrelin receptor antagonist [d-Lys3]GHRP-6 (1 μmol/kg), or various combinations of these compounds on portal and systemic levels of glucose and insulin after an intravenous glucose tolerance test (IVGTT, d-glucose 1 g/kg) in anesthetized fasted Wistar rats. UAG administration potently and dose-dependently enhanced the rise of insulin concentration induced by IVGTT in the portal and, to a lesser extent, the systemic circulation. This UAG-induced effect was completely blocked by the coadministration of exogenous AG at equimolar concentrations. Similarly to UAG, [d-Lys3]GHRP-6, alone or in combination with AG and UAG, strongly enhanced the portal insulin response to IVGTT, whereas exogenous AG alone did not exert any further effect. Our data demonstrate that, in glucose-stimulated conditions, exogenous UAG acts as a potent insulin secretagogue, whereas endogenous AG exerts a maximal tonic inhibition on glucose-induced insulin release.

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Aucamp, H. "Die skryf van kabaret- en liedtekste: Enkele praktiese wenke." Literator 21, no. 2 (2000): 81–88. http://dx.doi.org/10.4102/lit.v21i2.469.

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’n Werkwinkelsessie oor die skryf van kabaret mag nie verval in akademiese spekulasies nie, maar as jy wenke puntsgewyse wil deurgee, soos ek nou gaan doen, kan dit weer maklik lyk of die spreker voorskriftelik wil wees.

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Villemain, Jana L., and David P. Giedroc. "Characterization of a Cooperativity Domain Mutant Lys3→ Ala (K3A) T4 Gene 32 Protein." Journal of Biological Chemistry 271, no. 44 (1996): 27623–29. http://dx.doi.org/10.1074/jbc.271.44.27623.

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Grandjean, Frédéric, Marc Collas, Magali Uriarte, and Marion Rousset. "First record of a marbled crayfish Procambarus virginalis (Lyko, 2017) population in France." BioInvasions Records 10, no. 2 (2021): 341–47. http://dx.doi.org/10.3391/bir.2021.10.2.12.

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Scheers, Kevin, Rein Brys, Thomas Abeel, David Halfmaerten, Sabrina Neyrinck, and Tim Adriaens. "The invasive parthenogenetic marbled crayfish Procambarus virginalis Lyko, 2017 gets foothold in Belgium." BioInvasions Records 10, no. 2 (2021): 326–40. http://dx.doi.org/10.3391/bir.2021.10.2.11.

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