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Journal articles on the topic "MuT Proteins"
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Costanzo, Michele, Armando Cevenini, Emanuela Marchese, Esther Imperlini, Maddalena Raia, Luigi Del Vecchio, Marianna Caterino, and Margherita Ruoppolo. "Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line." International Journal of Molecular Sciences 19, no. 11 (November 13, 2018): 3580. http://dx.doi.org/10.3390/ijms19113580.
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Methylmalonic acidemias (MMAs) are inborn errors of metabolism due to the deficient activity of methylmalonyl-CoA mutase (MUT). MUT catalyzes the formation of succinyl-CoA from methylmalonyl-CoA, produced from propionyl-CoA catabolism and derived from odd chain fatty acids β-oxidation, cholesterol, and branched-chain amino acids degradation. Increased methylmalonyl-CoA levels allow for the presymptomatic diagnosis of the disease, even though no approved therapies exist. MMA patients show hyperammonemia, ketoacidosis, lethargy, respiratory distress, cognitive impairment, and hepatomegaly. The long-term consequences concern neurologic damage and terminal kidney failure, with little chance of survival. The cellular pathways affected by MUT deficiency were investigated using a quantitative proteomics approach on a cellular model of MUT knockdown. Currently, a consistent reduction of the MUT protein expression was obtained in the neuroblastoma cell line (SH-SY5Y) by using small-interfering RNA (siRNA) directed against an MUT transcript (MUT siRNA). The MUT absence did not affect the cell viability and apoptotic process in SH-SY5Y. In the present study, we evaluate and quantify the alterations in the protein expression profile as a consequence of MUT-silencing by a mass spectrometry-based label-free quantitative analysis, using two different quantitative strategies. Both quantitative methods allowed us to observe that the expression of the proteins involved in mitochondrial oxido-reductive homeostasis balance was affected by MUT deficiency. The alterated functional mitochondrial activity was observed in siRNA_MUT cells cultured with a propionate-supplemented medium. Finally, alterations in the levels of proteins involved in the metabolic pathways, like carbohydrate metabolism and lipid metabolism, were found.
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Stewart, Gavin S., Robert A. Fenton, Weidong Wang, Tae-Hwan Kwon, Stanley J. White, Valerie M. Collins, Gordon Cooper, Søren Nielsen, and Craig P. Smith. "The basolateral expression of mUT-A3 in the mouse kidney." American Journal of Physiology-Renal Physiology 286, no. 5 (May 2004): F979—F987. http://dx.doi.org/10.1152/ajprenal.00334.2003.
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Facilitative UT-A urea transporters play a central role in the urinary concentrating mechanism. There are three major UT-A isoforms found in the mouse kidney: mUT-A1, mUT-A2, and mUT-A3. The major aim of this study was to identify the location and function of mUT-A3. UT-A proteins were investigated using three novel mouse UT-A-targeted antibodies: ML446, MQ2, and ML194. ML446 detected mUT-A1 and mUT-A3. ML194 detected mUT-A1 and mUT-A2. Importantly, MQ2 was found to be selective for mUT-A3. MQ2 detected a 45- to 65-kDa signal in the mouse kidney inner medulla, which was deglycosylated to a 40-kDa protein band. Immunolocalization studies showed that mUT-A3 was strongly detected in the papillary tip, mainly in the basolateral regions of inner medullary collecting duct (IMCD) cells. Immunoblotting of subcellular fractions of inner medullary protein suggested that in mouse kidney mUT-A3 was present in plasma membranes. Consistent with this, immunoelectron microscopy demonstrated that mUT-A3 was predominantly localized at the basal plasma membrane domains of the IMCD cells in mouse kidney. Heterologous expression of mUT-A3-enhanced green fluorescent protein in Madin-Darby canine kidney cells showed that the protein localized to the basolateral membrane. In conclusion, our study indicates that mUT-A3 is a basolateral membrane transporter expressed in IMCD cells.
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Lewintre, Eloisa Jantus, Miguel Marin, Cristina Reinoso, David Montaner, Joaquín Dopazo, Juan Jose Calvete, and Javier Garcia-Conde. "Analysis of B-CLL Transcriptomic and Proteomic Profiles: Differences between Molecular Subgroups." Blood 108, no. 11 (November 1, 2006): 2088. http://dx.doi.org/10.1182/blood.v108.11.2088.2088.
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Abstract INTRODUCTION: B cell chronic lymphocytic leukemia (B-CLL) is a lymphoproliferative disorder with a variable clinical course. Patients (pts) with unmutated (unmut) IgVH gene show a shorter progression free survival and overall survival than the patients with IgVH mutated (mut). To understand the differences between molecular subgroups of B-CLL we have studied transcriptomic and proteomic profiles on samples from 40 B-CLL pts (Binet stage A). MATERIAL AND METHODS: 100 μg of total PBMC proteins were used for IEF followed by 2D electrophoresis. Image analysis of scanned gels was used to identify statistically differentially expressed proteins. Image acquisition and analysis were performed using the Ludesi software (http://www.ludesi.com). Selected spots were subjected to automatic digestion and the proteins were identified by MALDI-TOF (Voyager DE-Pro, Applied Biosystems) peptide mass fingerprint using the Protein Prospector software. To confirm the initial results, sequencing of selected peptide ions was carried out by collision-induced dissociation (CID) with a nESI-QTRAP mass spectrometer from Applied Biosystems. Eight proteins were validated by Western Blot. Total RNA from B cells was used to analyze the expression profile by hybridization with Whole Human Genome U133 Plus 2.0 Array from Affymetrix. Normalization, differential gene expression and functional annotations were analyzed using the GEPAS suite (http://www.gepas.org). qPCR using TaqMan primers/probes was used for validation of the microarray data RESULTS: When we compared IgVH mut vs unmut transcriptomic and proteomic profiles, we found more than 600 differentially expressed genes and 12 proteins ( fdr <0.05 adjusted for multiple test contrast and p<0.05, Mann-Whitney’s test, for gene and proteins, respectively). In tables 1 and 2 we show some of the most differentially expressed gene/proteins in each group of pts. Validation of results from microarrays and proteomic data using qPCR and Western blot are in progress. We obtained positive correlation between transcriptomic and proteomic profiles, (corr=0.21, p=0.04, Pearson’s correation test) suggesting that common features are found using both approximations. CONCLUSION: We found a number of interesting gene/proteins that could be able to differentiate molecular subgroups of B-CLL pts. The study of these proteins and genes may lead to better understand the different clinical behaviour of IgVH mut and unmut B-CLL forms, but validation with a larger group of pts is still necessary. Table 1: Genes differentially expressed IgVH mut IgVH unmut Genes annotated using their gene symbol BCL11A MGC9913 DUSP22 RGS4 PDLIM5 CRY1 RDH13 GGT2 PHF15 DMD SVH TUBB6 ADAM29 LPL ITPKB ITGA9 RBKS BCL7A NFATC1 MYEOV RIN3 PPP1R9A Table 2: Proteins differentially expressed IgVH mut IgVH unmut Proteins were annotated according to their gene symbol VIM ERP29 COTL1 CCT2 S100A9 PSMB10 HSPD1
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Pitocchi, Rossana, Paola Cicatiello, Leila Birolo, Alessandra Piscitelli, Elena Bovio, Giovanna Cristina Varese, and Paola Giardina. "Cerato-Platanins from Marine Fungi as Effective Protein Biosurfactants and Bioemulsifiers." International Journal of Molecular Sciences 21, no. 8 (April 21, 2020): 2913. http://dx.doi.org/10.3390/ijms21082913.
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Two fungal strains, Aspergillus terreus MUT 271 and Trichoderma harzianum MUT 290, isolated from a Mediterranean marine site chronically pervaded by oil spills, can use crude oil as sole carbon source. Herein, these strains were investigated as producers of biosurfactants, apt to solubilize organic molecules as a preliminary step to metabolize them. Both fungi secreted low molecular weight proteins identified as cerato-platanins, small, conserved, hydrophobic proteins, included among the fungal surface-active proteins. Both proteins were able to stabilize emulsions, and their capacity was comparable to that of other biosurfactant proteins and to commercially available surfactants. Moreover, the cerato-platanin from T. harzianum was able to lower the surface tension value to a larger extent than the similar protein from A. terreus and other amphiphilic proteins from fungi. Both cerato-platanins were able to make hydrophilic a hydrophobic surface, such as hydrophobins, and to form a stable layer, not removable even after surface washing. To the best of our knowledge, the ability of cerato-platanins to work both as biosurfactant and bioemulsifier is herein demonstrated for the first time.
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Costanzo, Michele, Marianna Caterino, Armando Cevenini, Vincent Jung, Cerina Chhuon, Joanna Lipecka, Roberta Fedele, Ida Chiara Guerrera, and Margherita Ruoppolo. "Proteomics Reveals that Methylmalonyl-CoA Mutase Modulates Cell Architecture and Increases Susceptibility to Stress." International Journal of Molecular Sciences 21, no. 14 (July 15, 2020): 4998. http://dx.doi.org/10.3390/ijms21144998.
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Methylmalonic acidemia (MMA) is a rare inborn error of metabolism caused by deficiency of the methylmalonyl-CoA mutase (MUT) enzyme. Downstream MUT deficiency, methylmalonic acid accumulates together with toxic metabolites from propionyl-CoA and other compounds upstream of the block in the enzyme pathway. The presentation is with life-threatening acidosis, respiratory distress, brain disturbance, hyperammonemia, and ketosis. Survivors develop poorly understood multi-organ damage, notably to the brain and kidneys. The HEK 293 cell line was engineered by CRISPR/Cas9 technology to knock out the MUT gene (MUT-KO). Shotgun label-free quantitative proteomics and bioinformatics analyses revealed potential damaging biological processes in MUT-deficient cells. MUT-KO induced alteration of cellular architecture and morphology, and ROS overproduction. We found the alteration of proteins involved in cytoskeleton and cell adhesion organization, cell trafficking, mitochondrial, and oxidative processes, as validated by the regulation of VIM, EXT2, SDC2, FN1, GLUL, and CHD1. Additionally, a cell model of MUT-rescuing was developed in order to control the specificity of MUT-KO effects. Globally, the proteomic landscape of MUT-KO suggests the cell model to have an increased susceptibility to propionate- and H2O2-induced stress through an impairment of the mitochondrial functionality and unbalances in the oxidation-reduction processes.
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Samodra, Eunike Marganingrum Andriani, Dian Suroto, Tyas Utami, Rachma Wikandari, and Endang Sutriswati Rahayu. "Cold Stress Response Genes of Lantiplantibacillus plantarum subsp. plantarum Mut-3 and Lantiplantibacillus plantarum subsp. plantarum Mut-7 Support the Ability to Survive in Low-Temperature Conditions." HAYATI Journal of Biosciences 30, no. 1 (August 19, 2022): 65–70. http://dx.doi.org/10.4308/hjb.30.1.65-70.
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Probiotics are widely consumed in various food matrices to provide health benefits to the host. The viability of probiotic cells is influenced by several factors, including exposure to high temperatures during the production process and low temperatures during storage. In this study, we report the response to cold stress of Lantiplantibacillus plantarum subsp. plantarum Mut-3 and Mut-7 after 24 h of storage at 4°C and -20℃. The cell number of Lantiplantibacillus plantarum subsp. plantarum Mut-3 and Mut-7 in low-temperature condition had no significant differences than their initial number: 11.88 log CFU/ml and 11.62 log CFU/ml at 4°C; 11.51 log CFU/ml and 11.47 log CFU/ml at -20°C for Mut-3 and Mut-7 respectively. The results indicated the survival capacity of Lantiplantibacillus plantarum subsp. plantarum Mut-3 and Mut-7 at low temperatures. The genes encoding cold shock proteins for the response to cold stress were evaluated by genome sequencing. The CspA/CspC genes of Lantiplantibacillus plantarum subsp. plantarum Mut-3 and Mut-7 possibly play a role in maintaining cell resistance at low temperatures, since the genes products predicted to have conserved motifs in the RNA binding protein (RNP) -1 and RNP-2 responsible for cold response stress which are similar to those in other bacteria.
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Chinsangaram, Jarasvech, Clayton Beard, Peter W. Mason, Marla K. Zellner, Gordon Ward, and Marvin J. Grubman. "Antibody Response in Mice Inoculated with DNA Expressing Foot-and-Mouth Disease Virus Capsid Proteins." Journal of Virology 72, no. 5 (May 1, 1998): 4454–57. http://dx.doi.org/10.1128/jvi.72.5.4454-4457.1998.
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ABSTRACT Candidate foot-and-mouth disease (FMD) DNA vaccines designed to produce viral capsids lacking infectious viral nucleic acid were evaluated. Plasmid DNAs containing a portion of the FMDV genome coding for the capsid precursor protein (P1-2A) and wild-type or mutant viral proteinase 3C (plasmids P12X3C or P12X3C-mut, respectively) were constructed. Cell-free translation reactions programmed with pP12X3C (wild-type 3C) and pP12X3C-mut produced a capsid precursor, but only the reactions programmed with the plasmid encoding the functional proteinase resulted in P1-2A processing and capsid formation. Baby hamster kidney (BHK) cells also produced viral capsid proteins when transfected with these plasmids. Plasmid P12X3C was administered to mice by intramuscular, intradermal, and epithelial (gene gun) inoculations. Anti-FMD virus (FMDV) antibodies were detected by radioimmunoprecipitation (RIP) and plaque reduction neutralization assays only in sera of mice inoculated by using a gene gun. When pP12X3C and pP12X3C-mut were inoculated into mice by using a gene gun, both plasmids elicited an antibody response detectable by RIP but only pP12X3C elicited a neutralizing antibody response. These results suggest that capsid formation in situ is required for effective immunization. Expression and stimulation of an immune response was enhanced by addition of an intron sequence upstream of the coding region, while addition of the FMDV internal ribosome entry site or leader proteinase (L) coding region either had no effect or reduced the immune response.
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Capaci, Valeria, Fiamma Mantovani, and Giannino Del Sal. "A mutant p53/Hif1α/miR-30d axis reprograms the secretory pathway promoting the release of a prometastatic secretome." Cell Stress 4, no. 11 (November 9, 2020): 261–64. http://dx.doi.org/10.15698/cst2020.11.235.
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TP53 missense mutations are frequent driver events during tumorigenesis. The majority of TP53 mutations are missense and occur within the DNA binding domain of p53, leading to expression of mutant p53 (mut-p53) proteins that not only lose the tumor suppressive functions of the wild-type (wt-p53) form, but can also acquire novel oncogenic features fostering tumor growth, metastasis and chemoresistance. Mut-p53 affects fundamental cellular pathways and functions through different mechanisms, a major one being the alteration of gene expression. In our recent work (Capaci et al., 2020, Nat Commun) we found that mut-p53, via miR-30d, modifies structure and function of the Golgi apparatus (GA) and induces increased rate of trafficking. This culminates in the release of a pro-malignant secretome, which is capable of remodeling the tumor microenvironment (TME), to increase stiffness of the extracellular matrix (ECM), favouring metastatic colonization, as shown by cell-based assays and experiments of metastatic niche preconditioning in mouse xenograft models. This study provides new insights into the mechanisms by which mut-p53, through induction of non-coding RNAs, can exert pro-tumorigenic functions in a non-cell-autonomous fashion, and highlights potential non-invasive biomarkers and therapeutic targets to treat tumors harboring mut-p53 (Figure 1).
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Tian, Bing, Yuhong Zhang, Bruce A. Luxon, Roberto P. Garofalo, Antonella Casola, Mala Sinha, and Allan R. Brasier. "Identification of NF-κB-Dependent Gene Networks in Respiratory Syncytial Virus-Infected Cells." Journal of Virology 76, no. 13 (July 1, 2002): 6800–6814. http://dx.doi.org/10.1128/jvi.76.13.6800-6814.2002.
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ABSTRACT Respiratory syncytial virus (RSV) is a mucosa-restricted virus that is a leading cause of epidemic respiratory tract infections in children. In epithelial cells, RSV replication activates nuclear translocation of the inducible transcription factor nuclear factor κB (NF-κB) through proteolysis of its cytoplasmic inhibitor, IκB. In spite of a putative role in mediating virus-inducible gene expression, the spectrum of NF-κB-dependent genes induced by RSV infection has not yet been determined. To address this, we developed a tightly regulated cell system expressing a nondegradable, epitope-tagged IκBα isoform (Flag-IκBα Mut) whose expression could be controlled by exogenous addition of nontoxic concentrations of doxycycline. Flag-IκBα Mut expression potently inhibited IκBα proteolysis, NF-κB binding, and NF-κB-dependent gene transcription in cells stimulated with the prototypical NF-κB-activating cytokine tumor necrosis factor alpha (TNF-α) and in response to RSV infection. High-density oligonucleotide microarrays were then used to profile constitutive and RSV-induced gene expression in the absence or presence of Flag-IκBα Mut. Comparison of these profiles revealed 380 genes whose expression was significantly changed by the dominant-negative NF-κB. Of these, 236 genes were constitutive (not RSV regulated), and surprisingly, only 144 genes were RSV regulated, representing numerically ∼10% of the total population of RSV-inducible genes at this time point. Hierarchical clustering of the 144 RSV- and Flag-IκBα Mut-regulated genes identified two discrete gene clusters. The first group had high constitutive expression, and its expression levels fell in response to RSV infection. In this group, constitutive mRNA expression was increased by Flag-IκBα Mut expression, and the RSV-induced decrease in expression was partly inhibited. In the second group, constitutive expression was very low (or undetectable) and, after RSV infection, expression levels strongly increased. In this group, NF-κB was required for RSV-inducible expression because Flag-IκBα Mut expression blocked their induction by RSV. This latter cluster includes chemokines, transcriptional regulators, intracellular proteins regulating translation and proteolysis, and secreted proteins (complement components and growth factor regulators). These data suggest that NF-κB action induces global cellular responses after viral infection.
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Dal Bo, Michele, Federico Pozzo, Riccardo Bomben, Antonella Zucchetto, Erika Tissino, Dania Benedetti, Massimo Degan, et al. "Nucleophosmin-1 and Ribosome-Associated Components Are Constitutively Overexpressed in NOTCH1 Mutated IGHV Unmutated CLL." Blood 120, no. 21 (November 16, 2012): 3880. http://dx.doi.org/10.1182/blood.v120.21.3880.3880.
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Abstract Abstract 3880 Background: activating mutations of NOTCH1 have been identified in about 10% of chronic lymphocytic leukemia (CLL) cases at diagnosis, with a higher frequency in unmutated IGHV (IGHV-UM) CLL, chemorefractory CLL and CLL in advanced disease phases. In CLL, all NOTCH1 mutations disrupt the C-terminal PEST domain and cause an accumulation of an active NOTCH1 isoform. Notably, about 80% of NOTCH1 mutations are represented by a CT frameshift deletion at nucleotides 7544–7545 (c.7544–7545delCT). Clinically, the presence of NOTCH1 mutations is an independent predictor of overall survival in CLL and identifies a subset of patients with particularly unfavourable prognosis (Rossi et al, Blood, 119, 521, 2012). Aim: to identify peculiar molecular and biological features of NOTCH1 mutated CLL in the context of IGHV-UM CLL. Methods: the presence of the c.7544–7545delCT NOTCH1 frameshift deletion was investigated by an ad-hoc amplification refractory mutation system (ARMS) PCR set up to obtain an amplicon specific for the NOTCH1 mutated form and a second amplicon as control. The percentage of NOTCH1 DNA in the context of the CLL clone was determined by quantitative real-time PCR (QRT-PCR), calculating the ratio between the amount of the specific NOTCH1 mutated amplicon and the amount of the control amplicon, the latter representing the total amount of NOTCH1 DNA irrespective of its mutational status. Gene expression profile (GEP) was performed by a one-color labeling strategy using the 4×44K Agilent platform. The differential expression of specific genes/proteins was validated by QRT-PCR, western blotting and immunohistochemistry. A BrdU uptake assay was used to evaluate proliferation of CLL cells by CpG/IL2 stimulation. Results: in a cohort of 380 IGHV-UM CLL, the c.7544–7545delCT NOTCH1 mutation was found in 83/380 (21.8%) cases. QRT-PCR revealed a percentage of NOTCH1 mutated DNA ranging from 1 to 37%. CLL cases carrying the c.7544–7545delCT NOTCH1 mutation (NOTCH1-Mut) showed higher NOTCH1 protein expression than CLL cases lacking NOTCH1-Mut employing monoclonal antibodies either recognizing the trans-membrane (mean fold increase=3) or the intra-citoplasmic (mean fold increase=2.1) NOTCH1 domain. A GEP comparing RNA from purified CLL samples of 5 NOTCH1-Mut CLL and 5 CLL lacking NOTCH1-Mut was performed, selecting the 5 NOTCH1-Mut cases among those with the higher percentages of NOTCH1 mutated DNA (percentages of NOTCH1 mutated DNA ranging from 15 to 37%). This approach selected the nucleophosmin 1 gene (NPM1) and genes codifying for several ribosomal proteins (RPS6, RPS10, RPS17, RPS28, RPSA, RPL7A, RPL18) as significantly up regulated in NOTCH1-Mut CLL cases. A higher expression of the above mentioned genes in NOTCH1-Mut CLL was validated in a wider series of 34 cases (18 NOTCH1-Mut cases; NPM1, p=0.03; RPS6, p=0.045; RPS10, p=0.048; RPS17, p=0.048; RPS28, p=0.049; RPSA, p=0.048; RPL7A, p=0.039; RPL18, p=0.041, respectively). Western blot analysis in 8 cases (4 NOTCH1-Mut cases) confirmed a higher NPM1 expression in NOTCH1-Mut cases (range of fold increase from 1.6 to 5.2) also at protein level. Consistently, lymph nodes preparations from NOTCH1-Mut CLL cases revealed a strong NPM1 staining both in nucleoli and cytoplasms. Finally, when stimulated in-vitro with the CpG/IL2 combination, NOTCH1-mut IGHV-UM CLL cells proliferated, as detected by a BrdU uptake assay (>10 fold increase over control), and up-regulated NPM1 both at transcript (mean fold increase=2.02 after 18 hours of CpG exposure, p=0.001) and protein (fold increase of 1.34 after 6 hours of CpG exposure) levels. Conclusion: NPM1 was identified as constitutively overexpressed in NOTCH1-Mut IGHV-UM CLL together with several ribosome-associated components. These findings are suggestive for an increased activity of the ribosomal machinery in NOTCH1-Mut IGHV-UM CLL as part of the molecular processes leading to control of CLL cell growth and survival in this clinically unfavourable disease subset. Disclosures: No relevant conflicts of interest to declare.
Dissertations / Theses on the topic "MuT Proteins"
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Hui, Daniel Jason. "The Mechanism of Protein Synthesis Inhibition by the P56 Family of Viral Stress Inducible Proteins." Connect to text online, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=case1104848977.
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Braute, Petter, and Jorg Eliassen Rødsjø. "Protein function prediction using annotated protein-protein interaction networks." Thesis, Norwegian University of Science and Technology, Department of Computer and Information Science, 2005. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-9177.
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Pereira, Joice Naiara Bertaglia. "Avaliação morfoquantitativa e ultraestrutural da cartilagem do processo condilar da mandíbula e da sincondrose basioccipital de ratos Wistar subnutridos e suplementados com resveratrol." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/10/10132/tde-16122015-121456/.
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A subnutrição é uma doença causada pela ingestão inadequada e/ou insuficiente de energia ou de outras biomoléculas, que pode comprometer o crescimento e o desenvolvimento dos tecidos. Em conjunto, a cartilagem do processo condilar da mandíbula e a sincondrose basioccipital representam os dois centros efetivos do crescimento craniofacial, que depende, dentre outros, de fatores locais como a produção do fator de crescimento semelhante à insulina I (IGF-I) e seu receptor (IGF-IR). Alterações tanto no peptídeo quanto no receptor estão associadas a graves atrasos no crescimento e desenvolvimento ósseo. Dentre os nutrientes alternativos com a capacidade de promoção da recuperação nutricional, destaca-se o resveratrol, um polifenol contido na uva e derivados que pode promover a melhora do crescimento através de propriedades osteogênicas, além de atuar como condroprotetor. Assim, a presente pesquisa teve por objetivo avaliar os efeitos da subnutrição proteica na cartilagem do processo condilar da mandíbula (PC) e na sincondrose basioccipital (SBo) de ratos, bem como os processos de renutrição e suplementação com resveratrol nos períodos pré-púbere (21 dias) e púbere (60 dias). Os grupos nutrido (N) e subnutrido (S) foram formados por filhotes cujas mães receberam dieta normoproteica e hipoproteica, respectivamente, desde o período de acasalamento; ao completarem 21 dias de vida extrauterina, época determinada para o desmame, os filhotes foram eutanasiados. A partir do 22° dia de vida, filhotes dos grupos N e S foram mantidos com as respectivas dietas até o 60° dia de vida quando constituíram, respectivamente, os grupos NN e SS. Dois grupos considerados renutridos foram formados por animais do grupo S, que a partir do 22° dia passaram a receber, o primeiro, a dieta normoproteica com 20% de caseína (grupo RN), e o segundo, a dieta normoproteica acrescida de resveratrol (RRv). A cartilagem do PC e a SBo foram processadas através das técnicas histológicas rotineiras e posteriormente submetidas às colorações da HE, Picrossíruis e Safranina-O, para a evidenciação dos componentes celular e colágeno e imunohistoquímica para a marcação de células reativas ao IGF-I e IGF-IR. Os aspectos ultraestruturais também foram analisados através da microscopia eletrônica de transmissão. Os resultados mostraram que a subnutrição foi capaz de alterar os componentes da matriz extracelular (MEC), acarretando em modificações na secreção das fibrilas colágenas e acúmulo de proteoglicanas nos tecidos; diminuição do número de células imunorreativas ao IGF-I e IGF-IR na cartilagem do PC e atraso no crescimento ósseo endocondral, comprovado através das avaliações quantitativas e morfométricas, em ambos os tecidos. A renutrição com dieta normal foi parcialmente eficaz na recuperação dos parâmetros avaliados, e pode ter provocado uma diminuição da capacidade de resposta celular ao IGF-I. Os efeitos condroprotetores do resveratrol foram fundamentados através da recuperação dos componentes da MEC e organelas dos condrócitos e uma possível diminuição da insensibilidade ao IGF-I; o atraso no crescimento ósseo endocondral foi minimizado através do seu potencial osteogênico (mais acentuado na SBo). Muito embora a recuperação dos tecidos não tenha ocorrido totalmente, diante dos dados apresentados, pode-se afirmar que a suplementação de resveratrol na dieta auxiliou de forma mais efetiva no restabelecimento dos parâmetros afetados pela subnutrição do que somente a recuperação proteica com dieta normalUndernutrition is a disease caused by inadequate and / or insufficient intake of energy or other biomolecules, which can compromise growth and development of tissues. Together, the mandibular condylar cartilage and basioccipital synchondrosis represent the two effective centers of craniofacial growth, which depends of the factors such as the local production of insulin like growth factor I (IGF-I) and receptor (IGF-IR). Changes in both the peptide and receptor are associated with serious delays in growth and bone development. Among the alternative nutrients to the promotion capacity of nutritional recovery, there is resveratrol, a polyphenol contained in grapes and derivatives that can promote improved growth through osteogenic properties, besides acting as chondroprotective. Thus, the present study aimed to evaluate in rats the effects of protein undernutrition in the mandibular condylar cartilage and basioccipital synchondrosis, as well renutrition and supplementation with resveratrol in pre pubertal periods (21 days) and pubertal (60 days). The nourished groups (N) and undernourished (S) were formed by young whose mothers received normal protein and hypoproteic diet respectively since the breeding season; to complete 21 days of extra-uterine life, given time to weaning, puppies were euthanized. From the 22nd day of life, puppies from N and S groups were kept with their diets until the 60th day of life when constituted, respectively, NN and SS groups. Two groups considered re-nourished were formed by the S group, which from day 22 began receiving, the first, the normal protein diet with 20% casein (RN group), and the second, the normal protein diet plus of resveratrol (RRv). The cartilage of PC and SBo were processed through routine histological techniques, and then subjected to HE, Sirius red and Safranin-O staining for the disclosure of cellular components and collagen and immunohistochemistry for marking cells responsive to IGF-I and IGF-IR. The ultrastructural aspects were also analyzed by transmission electron microscopy. The results showed that undernutrition was able to change the components of the extracellular matrix (ECM) leading to changes in the secretion of collagen fibrils and proteoglycans accumulation in the tissues; decrease in the number of immunoreactive cells to IGF-I and IGF-IR in PC cartilage and delayed endochondral bone growth, confirmed by quantitative and morphometric analysis in both tissues. Renutrition with normal diet was partially effective in recovering the parameters evaluated, and may have caused a decrease in cell responsiveness to IGF-I. The chondroprotective effects of resveratrol were founded by recovering the ECM components and organelles of chondrocytes and a possible decrease in the insensitivity to IGF-I; the delay in endochondral bone growth was minimized by osteogenic potential (more pronounced in SBo). Although the tissue recovery has not fully occurred, the resveratrol supplementation in the diet helped more effectively in restoring the parameters affected by undernutrition than just protein recovery with normal diet
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Treptow, Till. "Identifizierung und Charakterisierung zweier neuer Proteine, die mit dem HIV-Rev-Protein interagieren." Diss., lmu, 2009. http://nbn-resolving.de/urn:nbn:de:bvb:19-109373.
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Bardeleben, Anna Christina von. "Identifikation von neuen Schlitzmembranproteinen: NEPH-Proteine interagieren mit dem PDZ-Domänen-Protein PICK1." [S.l. : s.n.], 2008. http://nbn-resolving.de/urn:nbn:de:bsz:25-opus-55735.
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Maschkowitz, Gregor [Verfasser]. "Untersuchung der Protein-Protein-Interaktion der Proteine ppUL35 und ppUL35A des humanen Cytomegalievirus mit dem zellulären Protein Sorting Nexin 5 / Gregor Maschkowitz." Kiel : Universitätsbibliothek Kiel, 2017. http://d-nb.info/1138979988/34.
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Laborenz, Janina [Verfasser], and Johannes [Akademischer Betreuer] Herrmann. "Ema19 - ein ER-Protein mit Relevanz für mitochondriale Proteine / Janina Laborenz ; Betreuer: Johannes Herrmann." Kaiserslautern : Technische Universität Kaiserslautern, 2020. http://d-nb.info/1224474880/34.
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Franz, Martin [Verfasser]. "Vermehrung und Analyse des abnormalen Prion-Proteins mit Hilfe der protein misfolding cyclic amplification / Martin Franz." Greifswald : Universitätsbibliothek Greifswald, 2012. http://d-nb.info/1028232330/34.
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Schmidt, Michel. "Recombinant structural proteins of rubella virus." Helsinki : University of Helsinki, 2000. http://ethesis.helsinki.fi/julkaisut/mat/bioti/vk/schmidt/.
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Fu, Karen 1967. "Stability of proteins within biodegradable microspheres." Thesis, Massachusetts Institute of Technology, 2000. http://hdl.handle.net/1721.1/8422.
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Thesis (Sc.D.)--Harvard--Massachusetts Institute of Technology Division of Health Sciences and Technology, 2000.Includes bibliographical references.
In the past decade, biodegradable polymers have become the materials of choice for a variety of biomaterials applications. In particular, poly(lactic-co-glycolic acid) (PLGA) microspheres have been extensively studied for controlled-release protein delivery. However, significant issues arise in formulating such delivery systems since few proteins have been successfully encapsulated and released from these microspheres. Here, methods are developed to determine the causes of protein instability and solutions are provided for overcoming these problems. A commonly used technique for protein encapsulation in PLGA microspheres is the double-emulsion method. The harsh processing associated with this method can cause denaturation of the encapsulated protein. Herein, we have used Fourier transform infrared (FTIR) spectroscopy to determine the secondary structures of two model proteins, bovine serum albumin (BSA) and chicken egg-white lysozyme, within PLGA microspheres. This is a novel technique for in situ evaluation of proteins within microspheres and potentially a powerful and quick method for assessment of formulations. Results for both proteins indicate changes in structure upon entrapment within the microspheres. However, addition of the stabilizing excipient trehalose prevents the denaturing effects incurred during processing. In addition, BSA released from microspheres is improved by incorporation of trehalose. With microspheres made by double emulsion, there is often a large, initial burst of drug release upon injection. This results in inefficient use of therapy. To prevent this loss, a modified spontaneous emulsification method was explored.
(cont.) With this procedure both protein and polymer are soluble in a single solvent system thus avoiding creation of a water/solvent interface. The process was optimized for microsphere size and protein loading. Addition of a charged surfactant served to improve protein solubility and thus increase protein loading. In vitro and in vivo release kinetics showed a minimal burst, lower than that found with double emulsion microspheres, followed by sustained release. Upon injection of the microspheres in vivo, the PLGA microspheres begin to degrade. Degradation of the polymer generates acidic monomers, and acidification of the inner polymer environment is a central issue in the development of these devices for drug delivery. To quantitatively determine the intraparticle acidity, pH-sensitive fluorescent dyes were entrapped within the microspheres and imaged with confocal fluorescence microscopy. The technique allows visualization of the spatial and temporal distribution of pH within the degrading microspheres. The data indicate the formation of a very acidic environment within the particles with the minimum pH as low as 1.5. The images show a pH gradient, with the most acidic environment at the center of the spheres and higher pH near the edges, which is characteristic of diffusion-controlled release of the acidic degradation products. Strategies to avoid the accumulation of acidic monomers involve decreasing the diffusion distance of the degradation products by either decreasing the overall diameter of the microspheres or creating porous particles.
by Karen Fu.
Sc.D.
Books on the topic "MuT Proteins"
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Boros, Peter. Hepatocyte growth factor: The basic principles. Austin: R.G. Landes, 1999.
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Schmidt, Holger. NMR-Lösungsstruktur der humanen Hck SH3-Domäne im Komplex mit einem artifiziellen, hochoffinen Peptid-Liganden. Jülich: Forschungszentrum Jülich, Zentralbibliothek, 2006.
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Meissner, Daniel. Vergleichende Analyse der Sec- und Tat-abhängigen sekretorischen Proteingewinnung mit Gram-positiven Bakterien als Wirtsorganismen. Jülich: Forschungszentrum Jülich GmbH, Zentralbibliothek, 2006.
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Lee, Kyung-Hea. Untersuchungen zur Bilanzierung des Proteins sowie des Fructoselysins und des Lysinoalanins bei hitzebehandeltem Casein an Ratten mit der Homoarginin-Technik und an Menschen. Kiel: Selbstverlag des Instituts für Humanernährung und Lebensmittelkunde der Christian-Albrechts-Universität Kiel, 1992.
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Denmark. Skattestyrelsesloven: Met kommentarer. [Copenhagen]: Jurist- og økonomforbundet, 1988.
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Kreutzenbeck, Peter Johannes. Export von Proteinen mit Zwillingsarginin-Signalsequenzen über den Tat-Weg in Escherichia coli. Julich: Forschungszentrum Jülich, Zentralbibliothek, 2005.
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Buys, E. A. Met het EG-recht strijdige belastingstelsels en de rechtsbescherming van de burger: Op het grensvlak van nationaal en communautair recht. Arnhem: Gouda Quint, 1994.
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Tran, Thi Tuyen. Analyse der Bindungsspezifität der humanen Lck-SH3-Domäne anhand artifizieller und physiologischer Peptid-Liganden und strukturelle Charakterisierung dieser Peptide im Komplex mit SH3-Domänen. Jülich: Forschungszentrum Jülich, Zentralbibliothek, 2005.
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(Korea), Munhwa Sinmunsa. Chŏngbu chonghap minwŏn mit sosong pʻallyejip: Nodong, semu, kŏnsŏl, minsa, kasa, haengjŏng, hyŏngsa. Sŏul: Munhwa Sinmunsa, 1991.
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Reutter, Felix. Synthese von Lipopeptid-Immunmodulatoren an fester Phase und Identifizierung von Isoformen von Proteinen mit massenspektrometrischen Methoden. [S.l: s.n.], 1999.
Find full textBook chapters on the topic "MuT Proteins"
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Zhang, Baichuan, Luying He, Qi Wang, Zhuo Wang, Wenzheng Bao, and Honglin Cheng. "Mit Protein Transformer: Identification Mitochondrial Proteins with Transformer Model." In Lecture Notes in Computer Science, 607–16. Singapore: Springer Nature Singapore, 2023. http://dx.doi.org/10.1007/978-981-99-4749-2_52.
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He, Wenliang, Peng Li, and Guoyao Wu. "Amino Acid Nutrition and Metabolism in Chickens." In Advances in Experimental Medicine and Biology, 109–31. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-54462-1_7.
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AbstractBoth poultry meat and eggs provide high-quality animal protein [containing sufficient amounts and proper ratios of amino acids (AAs)] for human consumption and, therefore, play an important role in the growth, development, and health of all individuals. Because there are growing concerns about the suboptimal efficiencies of poultry production and its impact on environmental sustainability, much attention has been paid to the formulation of low-protein diets and precision nutrition through the addition of low-cost crystalline AAs or alternative sources of animal-protein feedstuffs. This necessitates a better understanding of AA nutrition and metabolism in chickens. Although historic nutrition research has focused on nutritionally essential amino acids (EAAs) that are not synthesized or are inadequately synthesized in the body, increasing evidence shows that the traditionally classified nutritionally nonessential amino acids (NEAAs), such as glutamine and glutamate, have physiological and regulatory roles other than protein synthesis in chicken growth and egg production. In addition, like other avian species, chickens do not synthesize adequately glycine or proline (the most abundant AAs in the body but present in plant-source feedstuffs at low content) relative to their nutritional and physiological needs. Therefore, these two AAs must be sufficient in poultry diets. Animal proteins (including ruminant meat & bone meal and hydrolyzed feather meal) are abundant sources of both glycine and proline in chicken nutrition. Clearly, chickens (including broilers and laying hens) have dietary requirements for all proteinogenic AAs to achieve their maximum productivity and maintain optimum health particularly under adverse conditions such as heat stress and disease. This is a paradigm shift in poultry nutrition from the 70-year-old “ideal protein” concept that concerned only about EAAs to the focus of functional AAs that include both EAAs and NEAAs.
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Hundleby, Penny A. C., Marc-André D’Aoust, Carolyn Finkle, Judith Atkins, and Richard M. Twyman. "Regulation of Molecular Farming Products." In Recombinant Proteins in Plants, 313–33. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2241-4_17.
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AbstractThe regulation of molecular farming is a complex topic because plants and plant-based systems are relative newcomers among the many production platforms available for recombinant proteins. The regulations specific for different types of product (human/veterinary pharmaceuticals and medical devices, cosmetics, diagnostics, and research reagents) must therefore be overlaid with the regulations governing hitherto unfamiliar production platforms, and this must be achieved in different jurisdictions that handle genetically modified organisms (and genetically modified plants in particular) in very different ways. This chapter uses examples of different product types and production methods in three different jurisdictions (the USA, the EU, and Canada) to demonstrate some of the challenges facing the regulatory authorities.
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Santoro, Massimo Mattia, and Giovanni Gaudino. "The Constitutive Activation of MET, RON and SEA Genes Induces Different Biological Responses." In Interacting Protein Domains, 207–12. Berlin, Heidelberg: Springer Berlin Heidelberg, 1997. http://dx.doi.org/10.1007/978-3-642-60848-3_30.
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Bartoschik, Tanja, Andreas Zoephel, Klaus Rumpel, Alessio Ciulli, and Charles Heffern. "MST and Technology to Reliably Study Binary and Ternary Binding in Drug Development." In Targeted Protein Degradation, 115–33. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1665-9_6.
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Krause, Jens-Peter. "Coating mit pflanzlichen Proteinen." In Easy Coating, 165–78. Wiesbaden: Vieweg+Teubner, 2011. http://dx.doi.org/10.1007/978-3-8348-9896-8_8.
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Amirai, J., C. Boyer, Ch Rothschild, and M. Wolf. "Protein C-Bestimmung mit einem Enzymimmunoassay." In Protein C - Klinische Bedeutung und Bestimmungsmethoden, edited by Irene Witt and Ernst Zimmer, 17–30. Berlin, Boston: De Gruyter, 1986. http://dx.doi.org/10.1515/9783110888461-003.
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Auberger, K., H. Engelmann, J. Weil, Ch Brückmann, B. Dietz, W. Schramm, Th Vukovich, and H. B. Hadorn. "Substitutionstherapie mit hochgereinigtem Protein C-Konzentrat bei einem Kind mit homozygotem Protein C-Mangel." In 17. Hämophilie-Symposion, 364–66. Berlin, Heidelberg: Springer Berlin Heidelberg, 1988. http://dx.doi.org/10.1007/978-3-642-72830-3_85.
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Hellstern, P., G. von Blohn, C. Miyashita, M. Köhler, P. Scheffler, and E. Wenzel. "Plasma-Protein C-Spiegel unter Langzeitantikoagulation mit Phenprocoumon." In Protein C - Klinische Bedeutung und Bestimmungsmethoden, edited by Irene Witt and Ernst Zimmer, 119–24. Berlin, Boston: De Gruyter, 1986. http://dx.doi.org/10.1515/9783110888461-013.
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Eckert, Werner A., and Jürgen Kartenbeck. "Nachweis von Proteinen auf Blot-Membranen mit immunchemischen Methoden." In Proteine: Standardmethoden der Molekular- und Zellbiologie, 167–223. Berlin, Heidelberg: Springer Berlin Heidelberg, 1997. http://dx.doi.org/10.1007/978-3-642-59227-0_4.
Full textConference papers on the topic "MuT Proteins"
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Ferreira, Bianca Nicoletti, João Henrique Do Nascimento E. Silva, Silvia Raquel Bettani, Paula Porrelli Moreira Da Silva, and Marta Regina Verruma-Bernardi. "AVALIAÇÃO FÍSICO-QUÍMICA E SENSORIAL DE CAFÉS TORRADO E MOÍDO TRADICIONAL E EXTRAFORTE." In II Congresso Brasileiro de Biotecnologia On-line. Revista Multidisciplinar de Educação e Meio Ambiente, 2022. http://dx.doi.org/10.51189/conbiotec/28.
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Introdução: Cada vez mais os atributos qualitativos dos produtos são importantes para os consumidores, desta maneira a determinação dos componentes físico-químicos do grão de café beneficiado apresenta papel importante na qualidade da bebida de café. Objetivo: deste trabalho foi avaliar as características físico-químicas e sensoriais de marcas comerciais de café torrado e moído. Material e Métodos: Foram avaliadas quatro marcas (M) de cafés, comercializados como tradicional (T) e extraforte (EF): M1T; M1EF; M2T; M2EF; M3T; M3EF; M4T; M4EF. Os cafés foram analisados quanto à atividade de água, cor instrumental (Luminosidade), umidade, lipídeos, proteínas, cinzas, sólidos solúveis totais, pH, açúcares redutores e não redutores. O teste sensorial utilizado foi o de ordenação de diferença e preferência com 30 avaliadores. Os dados das análises físico-químicas foram submetidos a análise de variância e teste de Tukey e para os dados do teste de ordenação utilizou-se o teste de Friedman e em ambos foi utilizado o nível de significância p≤0,05. Resultados: Os resultados obtidos de atividade de água variaram entre 0,44 e 0,49, umidade 3,01 e 3,69% e sólidos solúveis 24 e 36%. Para os teores de açúcares redutores a variação foi de 0,8 a 1,23% e açúcares totais de 2,53 a 3,44%. Os: lipídeos variaram de 11,88 a 14,24%; proteínas 12,30 a 12,97%; cinzas 4,78 a 5,15%; pH 5,46 a 5,73 e açúcares não redutores 1,43 a 2,50%. Quanto à Luminosidade houve uma variação de 35,27 - 36,05. Na análise sensorial foi verificado que o café M1T caracterizou-se como a amostra mais clara e de menor preferência, confirmado na análise de Luminosidade. Conclusão: Assim, apesar dos cafés estudados apresentarem um processo de torra eficiente do ponto de vista de atividade de água e teor de umidade, a presença de variações entre alguns dos parâmetros analisados contribui para a caracterização heterogênea dos cafés comerciais.
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Kubinski, S., N. Hensel, R. Lindner, and P. Claus. "Amyotrophe Lateralsklerose (ALS)-assoziierte Proteine ko-lokalisieren mit dem Aktin-bindenden Protein Profilin." In 24. Kongress des Medizinisch-Wissenschaftlichen Beirates der Deutschen Gesellschaft für Muskelkranke (DGM) e.V. Georg Thieme Verlag KG, 2019. http://dx.doi.org/10.1055/s-0039-1685049.
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Schmitt, H., A. Warnecke, I. De Vries, T. Lenarz, S. Alvi, N. Prenzler, M. Durisin, and H. Staecker. "Charakterisierung des Proteoms humaner Perilymphe mit dem Focus auf BDNF regulierte Proteine." In Abstract- und Posterband – 89. Jahresversammlung der Deutschen Gesellschaft für HNO-Heilkunde, Kopf- und Hals-Chirurgie e.V., Bonn – Forschung heute – Zukunft morgen. Georg Thieme Verlag KG, 2018. http://dx.doi.org/10.1055/s-0038-1640584.
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Fuhlendorff, J., I. Clemmensen, and S. Magnusson. "PRIMARY STRUCTURE OF TETRANECTIN. SEQUENCE HOMOLOGY WITH ASIALOGLYCOPROTEIN RECEPTORS AND WITH PROTEOGLYCAN CORE PROTEIN FROM CARTILAGE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644380.
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Tetranectin (Mr = 68,000) is a tetrameric blood plasma protein, which binds to plasminogen and also to the lysine-binding site of the isolated kringle 4 from plasminogen. Its four polypeptide chains, which are non-covalently bound, each consists of 181 amino acid residues. We have determined the complete amino acid sequence and the disulfide bonds. Each position corresponds to a single amino acid residue except 34 which contains Ala and Ser and 37 which contains Val and Met in equimolar amounts. The three disulfide bonds connect Cys-50 to Cys-60, Cys-77 to Cys-176 and Cys-152 to Cys-168. The sequence of tetranectin was found to be homologous, to an extent indicating common ancestry, with the extracellular part of the asialoglyco-protein receptors and with the C-terminal globular domain of the cartilage proteoglycan core protein. Conserved residues include the six half-cystines of tetranectin. Therefore, we can now propose disulfide bond patterns for the proteins homologous with tetranectin. Supported by NIH-grant HL-16238 (S.M.), the Danish Science and Medical Research Councils, the Danish Cancer Society and NOVO Foundation.
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Dayal, V. K., A. Hsieh, O. Velasquez, and Y. S. Arkel. "VONWILLEBRAND FRAGMENTS IN CoMMERCIAL FVIII CONCENTRATES." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644053.
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We have previously reported sane of the biochemical character-istics of varioias aomnercial FVIII concentrates. Two-dimensional gel analysis of these preparations indicated the presence of pep-tides in the 30-50 KDa basic region. We also noted a relative decrease of high molecular weight vonWillebrand factor (vWF) multimers on SDS-agarose gels. However, the proportion of low molecular weight vWF fragments seems to be increased as judged by the following studies. After reduction and electrophoresis in SDS- PAGE gels, the proteins in concentrates were electroblotted on to the nitrocellulose membranes. The vWF fragments were detected using sandwich of rabbit anti-human vWF antibody, followed by goat anti-rabbit antibody-horseradish peroxidase conjugate incubation and color development. These studies indicated the ore- senceof several vWF peptides in the 30-50 KDa region of one of the FVIII concentrates. These fragments were not detectable in normal plasma analyzed under identical conditions. In order to investigate any clinical significance of the 30-50 KDa peotides, the following experiments were carried out. A factor VIII concentrate preparation was subjected to electrophoresis in SDS-PAGE gels. After electrobiotting, the proteins nitrocellulose membrane were blocked and then reacted with hemoohilia a plasma. Subsequent reaction with 125I Protein A and autoradiograohy revealed the presence of a peptide in approximately 35 KDa region. A protein band of similar molecular weight also reacted with anti-vWF antibody in a separate identical run. Although the patient plasma did contain an antibody to a 35 KDa peotide, it is not certain whether this peptide is identical to the 35 KDa vWF fragment. The commercial FVIII concentrates that contain 30-59 KDa peptides also demonstrate a relative decrease in high molecular weight vWF mul timers. The possible role of proteolysis in the generation of vWF fragments is under study. The results reported here provide evidence that some of the conmercial FVIII concentrates contain peptides in 30-50 KDa region which are not detectable in normal plasma. Furthermore, sane of these low molecular weight peptides are attributable to vWF fragments.
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Weil, J., P. Maharjan, A. Beitia, K. Hilton, N. Suesuttajit, V. Naranjo, and C. N. Coon. "Responses of varying levels of DL-methionine and TSAA and effects on [13C]Met and [13C]Cys oxidation." In 6th EAAP International Symposium on Energy and Protein Metabolism and Nutrition. The Netherlands: Wageningen Academic Publishers, 2019. http://dx.doi.org/10.3920/978-90-8686-891-9_78.
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Jiang, Keren, Di Zhang, Tsubasa Iino, Risa Kimura, Tatsuo Nakajima, Kana Shimizu, Masahito Ohue, and Yutaka Akiyama. "A playful tool for predicting protein-protein docking." In MUM 2019: 18th International Conference on Mobile and Ubiquitous Multimedia. New York, NY, USA: ACM, 2019. http://dx.doi.org/10.1145/3365610.3368409.
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Meytin, Sophia. "A Novel Method for Protein-Protein Interface Analysis Using Sonification." In 2021 IEEE MIT Undergraduate Research Technology Conference ((URTC)). IEEE, 2021. http://dx.doi.org/10.1109/urtc54388.2021.9701622.
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Palmer, M. "Monocarboxylate Transporter (MCT) Proteins as Targets for Radiation-Induced Pulmonary Fibrosis." In American Thoracic Society 2019 International Conference, May 17-22, 2019 - Dallas, TX. American Thoracic Society, 2019. http://dx.doi.org/10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a5338.
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Burg, C. M., P. Backhaus, J. Tio, D. Neri, S. Cazzamalli, W. Heindel, and M. Schäfers. "Erste Erfahrungen mit dem neuen Liganden 68Ga-OncoFAP zur Darstellung des Fibroblasten-Aktivierungs-Protein (FAP) im Brustkrebs-Staging mittels PET-MRT." In 103. Deutscher Röntgenkongress der Deutschen Röntgengesellschaft e. V. Georg Thieme Verlag, 2022. http://dx.doi.org/10.1055/s-0042-1749847.
Full textReports on the topic "MuT Proteins"
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Bercovier, Herve, Raul Barletta, and Shlomo Sela. Characterization and Immunogenicity of Mycobacterium paratuberculosis Secreted and Cellular Proteins. United States Department of Agriculture, January 1996. http://dx.doi.org/10.32747/1996.7573078.bard.
Full textAbstract:
Our long-term goal is to develop an efficient acellular vaccine against paratuberculosis based on protein antigen(s). A prerequisite to achieve this goal is to analyze and characterize Mycobacterium paratuberculosis (Mpt) secreted and cellular proteins eliciting a protective immune response. In the context of this general objective, we proposed to identify, clone, produce, and characterize: the Mpt 85B antigen and other Mpt immunoreactive secreted proteins, the Mpt L7/L12 ribosomal protein and other immunoreactive cellular proteins, Mpt protein determinants involved in invasion of epithelial cells, and Mpt protein antigens specifically expressed in macrophages. Paratuberculosis is still a very serious problem in Israel and in the USA. In the USA, a recent survey evaluated that 21.6% of the dairy herd were infected with Mpt resulting in 200-250 million dollars in annual losses. Very little is known on the virulence factors and on protective antigens of Mpt. At present, the only means of controlling this disease are culling or vaccination. The current vaccines do not allow a clear differentiation between infected and vaccinated animals. Our long-term goal is to develop an efficient acellular paratuberculosis vaccine based on Mpt protein antigen(s) compatible with diagnostic tests. To achieve this goal it is necessary to analyze and characterize secreted and cellular proteins candidate for such a vaccine. Representative Mpt libraries (shuttle plasmid and phage) were constructed and used to study Mpt genes and gene products described below and will be made available to other research groups. In addition, two approaches were performed which did not yield the expected results. Mav or Mpt DNA genes that confer upon Msg or E. coli the ability to invade and/or survive within HEp-2 cells were not identified. Likewise, we were unable to characterize the 34-39 kDa induced secreted proteins induced by stress factors due to technical difficulties inherent to the complexity of the media needed to support substantial M. pt growth. We identified, isolated, sequenced five Mpt proteins and expressed four of them as recombinant proteins that allowed the study of their immunological properties in sensitized mice. The AphC protein, found to be up regulated by low iron environment, and the SOD protein are both involved in protecting mycobacteria against damage and killing by reactive oxygen (Sod) and nitrogen (AhpC) intermediates, the main bactericidal mechanisms of phagocytic cells. SOD and L7/L12 ribosomal proteins are structural proteins constitutively expressed. 85B and CFP20 are both secreted proteins. SOD, L7/L12, 85B and CFP20 were shown to induce a Th1 response in immunized mice whereas AphC was shown by others to have a similar activity. These proteins did not interfere with the DTH reaction of naturally infected cows. Cellular immunity provides protection in mycobacterial infections, therefore molecules inducing cellular immunity and preferentially a Th1 pathway will be the best candidate for the development of an acellular vaccine. The proteins characterized in this grant that induce a cell-mediated immunity and seem compatible with diagnostic tests, are good candidates for the construction of a future acellular vaccine.
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Epel, Bernard L., Roger N. Beachy, A. Katz, G. Kotlinzky, M. Erlanger, A. Yahalom, M. Erlanger, and J. Szecsi. Isolation and Characterization of Plasmodesmata Components by Association with Tobacco Mosaic Virus Movement Proteins Fused with the Green Fluorescent Protein from Aequorea victoria. United States Department of Agriculture, September 1999. http://dx.doi.org/10.32747/1999.7573996.bard.
Full textAbstract:
The coordination and regulation of growth and development in multicellular organisms is dependent, in part, on the controlled short and long-distance transport of signaling molecule: In plants, symplastic communication is provided by trans-wall co-axial membranous tunnels termed plasmodesmata (Pd). Plant viruses spread cell-to-cell by altering Pd. This movement scenario necessitates a targeting mechanism that delivers the virus to a Pd and a transport mechanism to move the virion or viral nucleic acid through the Pd channel. The identity of host proteins with which MP interacts, the mechanism of the targeting of the MP to the Pd and biochemical information on how Pd are alter are questions which have been dealt with during this BARD project. The research objectives of the two labs were to continue their biochemical, cellular and molecular studies of Pd composition and function by employing infectious modified clones of TMV in which MP is fused with GFP. We examined Pd composition, and studied the intra- and intercellular targeting mechanism of MP during the infection cycle. Most of the goals we set for ourselves were met. The Israeli PI and collaborators (Oparka et al., 1999) demonstrated that Pd permeability is under developmental control, that Pd in sink tissues indiscriminately traffic proteins of sizes of up to 50 kDa and that during the sink to source transition there is a substantial decrease in Pd permeability. It was shown that companion cells in source phloem tissue export proteins which traffic in phloem and which unload in sink tissue and move cell to cell. The TAU group employing MP:GFP as a fluorescence probe for optimized the procedure for Pd isolation. At least two proteins kinases found to be associated with Pd isolated from source leaves of N. benthamiana, one being a calcium dependent protein kinase. A number of proteins were microsequenced and identified. Polyclonal antibodies were generated against proteins in a purified Pd fraction. A T-7 phage display library was created and used to "biopan" for Pd genes using these antibodies. Selected isolates are being sequenced. The TAU group also examined whether the subcellular targeting of MP:GFP was dependent on processes that occurred only in the presence of the virus or whether targeting was a property indigenous to MP. Mutant non-functional movement proteins were also employed to study partial reactions. Subcellular targeting and movement were shown to be properties indigenous to MP and that these processes do not require other viral elements. The data also suggest post-translational modification of MP is required before the MP can move cell to cell. The USA group monitored the development of the infection and local movement of TMV in N. benthamiana, using viral constructs expressing GFP either fused to the MP of TMV or expressing GFP as a free protein. The fusion protein and/or the free GFP were expressed from either the movement protein subgenomic promoter or from the subgenomic promoter of the coat protein. Observations supported the hypothesis that expression from the cp sgp is regulated differently than expression from the mp sgp (Szecsi et al., 1999). Using immunocytochemistry and electron microscopy, it was determined that paired wall-appressed bodies behind the leading edge of the fluorescent ring induced by TMV-(mp)-MP:GFP contain MP:GFP and the viral replicase. These data suggest that viral spread may be a consequence of the replication process. Observation point out that expression of proteins from the mp sgp is temporary regulated, and degradation of the proteins occurs rapidly or more slowly, depending on protein stability. It is suggested that the MP contains an external degradation signal that contributes to rapid degradation of the protein even if expressed from the constitutive cp sgp. Experiments conducted to determine whether the degradation of GFP and MP:GFP was regulated at the protein or RNA level, indicated that regulation was at the protein level. RNA accumulation in infected protoplast was not always in correlation with protein accumulation, indicating that other mechanisms together with RNA production determine the final intensity and stability of the fluorescent proteins.
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Elbaum, Michael, and Peter J. Christie. Type IV Secretion System of Agrobacterium tumefaciens: Components and Structures. United States Department of Agriculture, March 2013. http://dx.doi.org/10.32747/2013.7699848.bard.
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Objectives: The overall goal of the project was to build an ultrastructural model of the Agrobacterium tumefaciens type IV secretion system (T4SS) based on electron microscopy, genetics, and immunolocalization of its components. There were four original aims: Aim 1: Define the contributions of contact-dependent and -independent plant signals to formation of novel morphological changes at the A. tumefaciens polar membrane. Aim 2: Genetic basis for morphological changes at the A. tumefaciens polar membrane. Aim 3: Immuno-localization of VirB proteins Aim 4: Structural definition of the substrate translocation route. There were no major revisions to the aims, and the work focused on the above questions. Background: Agrobacterium presents a unique example of inter-kingdom gene transfer. The process involves cell to cell transfer of both protein and DNA substrates via a contact-dependent mechanism akin to bacterial conjugation. Transfer is mediated by a T4SS. Intensive study of the Agrobacterium T4SS has made it an archetypal model for the genetics and biochemistry. The channel is assembled from eleven protein components encoded on the B operon in the virulence region of the tumor-inducing plasmid, plus an additional coupling protein, VirD4. During the course of our project two structural studies were published presenting X-ray crystallography and three-dimensional reconstruction from electron microscopy of a core complex of the channel assembled in vitro from homologous proteins of E. coli, representing VirB7, VirB9, and VirB10. Another study was published claiming that the secretion channels in Agrobacterium appear on helical arrays around the membrane perimeter and along the entire length of the bacterium. Helical arrangements in bacterial membranes have since fallen from favor however, and that finding was partially retracted in a second publication. Overall, the localization of the T4SS within the bacterial membranes remains enigmatic in the literature, and we believe that our results from this project make a significant advance. Summary of achievements : We found that polar inflations and other membrane disturbances relate to the activation conditions rather than to virulence protein expression. Activation requires low pH and nutrient-poor medium. These stress conditions are also reflected in DNA condensation to varying degrees. Nonetheless, they must be considered in modeling the T4SS as they represent the relevant conditions for its expression and activity. We identified the T4SS core component VirB7 at native expression levels using state of the art super-resolution light microscopy. This marker of the secretion system was found almost exclusively at the cell poles, and typically one pole. Immuno-electron microscopy identified the protein at the inner membrane, rather than at bridges across the inner and outer membranes. This suggests a rare or transient assembly of the secretion-competent channel, or alternatively a two-step secretion involving an intermediate step in the periplasmic space. We followed the expression of the major secreted effector, VirE2. This is a single-stranded DNA binding protein that forms a capsid around the transferred oligonucleotide, adapting the bacterial conjugation to the eukaryotic host. We found that over-expressed VirE2 forms filamentous complexes in the bacterial cytoplasm that could be observed both by conventional fluorescence microscopy and by correlative electron cryo-tomography. Using a non-retentive mutant we observed secretion of VirE2 from bacterial poles. We labeled the secreted substrates in vivo in order detect their secretion and appearance in the plant cells. However the low transfer efficiency and significant background signal have so far hampered this approach.
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Epel, Bernard, and Roger Beachy. Mechanisms of intra- and intercellular targeting and movement of tobacco mosaic virus. United States Department of Agriculture, November 2005. http://dx.doi.org/10.32747/2005.7695874.bard.
Full textAbstract:
To cause disease, plant viruses must replicate and spread locally and systemically within the host. Cell-to-cell virus spread is mediated by virus-encoded movement proteins (MPs), which modify the structure and function of plasmodesmata (Pd), trans-wall co-axial membranous tunnels that interconnect the cytoplasm of neighboring cells. Tobacco mosaic virus (TMV) employ a single MP for cell- cell spread and for which CP is not required. The PIs, Beachy (USA) and Epel (Israel) and co-workers, developed new tools and approaches for study of the mechanism of spread of TMV that lead to a partial identification and molecular characterization of the cellular machinery involved in the trafficking process. Original research objectives: Based on our data and those of others, we proposed a working model of plant viral spread. Our model stated that MPᵀᴹⱽ, an integral ER membrane protein with its C-terminus exposed to the cytoplasm (Reichel and Beachy, 1998), alters the Pd SEL, causes the Pd cytoplasmic annulus to dilate (Wolf et al., 1989), allowing ER to glide through Pd and that this gliding is cytoskeleton mediated. The model claimed that in absence of MP, the ER in Pd (the desmotubule) is stationary, i.e. does not move through the Pd. Based on this model we designed a series of experiments to test the following questions: -Does MP potentiate ER movement through the Pd? - In the presence of MP, is there communication between adjacent cells via ER lumen? -Does MP potentiate the movement of cytoskeletal elements cell to cell? -Is MP required for cell-to-cell movement of ER membranes between cells in sink tissue? -Is the binding in situ of MP to RNA specific to vRNA sequences or is it nonspecific as measured in vitro? And if specific: -What sequences of RNA are involved in binding to MP? And finally, what host proteins are associated with MP during intracellular targeting to various subcellular targets and what if any post-translational modifications occur to MP, other than phosphorylation (Kawakami et al., 1999)? Major conclusions, solutions and achievements. A new quantitative tool was developed to measure the "coefficient of conductivity" of Pd to cytoplasmic soluble proteins. Employing this tool, we measured changes in Pd conductivity in epidermal cells of sink and source leaves of wild-type and transgenic Nicotiana benthamiana (N. benthamiana) plants expressing MPᵀᴹⱽ incubated both in dark and light and at 16 and 25 ᵒC (Liarzi and Epel, 2005 (appendix 1). To test our model we measured the effect of the presence of MP on cell-to-cell spread of a cytoplasmic fluorescent probe, of two ER intrinsic membrane protein-probes and two ER lumen protein-probes fused to GFP. The effect of a mutant virus that is incapable of cell-to-cell spread on the spread of these probes was also determined. Our data shows that MP reduces SEL for cytoplasmic molecules, dilates the desmotubule allowing cell-cell diffusion of proteins via the desmotubule lumen and reduces the rate of spread of the ER membrane probes. Replicase was shown to enhance cell-cell spread. The data are not in support of the proposed model and have led us to propose a new model for virus cell-cell spread: this model proposes that MP, an integral ER membrane protein, forms a MP:vRNAER complex and that this ER-membrane complex diffuses in the lipid milieu of the ER into the desmotubule (the ER within the Pd), and spreads cell to cell by simple diffusion in the ER/desmotubule membrane; the driving force for spread is the chemical potential gradient between an infected cell and contingent non-infected neighbors. Our data also suggests that the virus replicase has a function in altering the Pd conductivity. Transgenic plant lines that express the MP gene of the Cg tobamovirus fused to YFP under the control the ecdysone receptor and methoxyfenocide ligand were generated by the Beachy group and the expression pattern and the timing and targeting patterns were determined. A vector expressing this MPs was also developed for use by the Epel lab . The transgenic lines are being used to identify and isolate host genes that are required for cell-to-cell movement of TMV/tobamoviruses. This line is now being grown and to be employed in proteomic studies which will commence November 2005. T-DNA insertion mutagenesis is being developed to identify and isolate host genes required for cell-to-cell movement of TMV.
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Amir, Rachel, David J. Oliver, Gad Galili, and Jacline V. Shanks. The Role of Cysteine Partitioning into Glutathione and Methionine Synthesis During Normal and Stress Conditions. United States Department of Agriculture, January 2013. http://dx.doi.org/10.32747/2013.7699850.bard.
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The objective of this research is to study the nature of the competition for cysteine (Cys), the first organic sulfur-containing compound, between its two main metabolites, glutathione (GSH) and methionine (Met). GSH plays a central role in protecting plants during various stresses, while Met, an essential amino acid, regulates essential processes and metabolites in plant cells through its metabolite S-adenosyl-Met. Our results, which are based on flux analysis and measurements of Met- metabolites, show that the flux towards Met synthesis is high during non-stress conditions, however the flux is significantly reduced under stress conditions, when there is high synthesis of GSH. Under oxidative stress the expression level of the regulatory enzyme of Met synthesis, cystathionine g-synthase (CGS) was reduced. By using three different systems, we have found that that GSH down regulates the expression level of CGS, thus reducing Met synthesis. We have found that this regulation occurs at the post-transcriptional level, and further studies have shown that it occurs at post-translationaly. To reveal how oxidative stress affects the flux towards Met and GSH, flux analysis was performed. We have found that the level of Met is significantly reduced, while the level of glutathione significantly increases during stress. Under stress conditions most of the glutathione is converted from GSH to GSSG (the oxidised form of glutathione). These results suggest that under normal growth conditions, Cys is channelled towards both pathways to support GSH accumulation and the synthesis of growth-essential Met metabolites. However, during oxidative stress, when a high level of GSH is required to protect the plants, the levels of GSH increase while those of CGS are reduced. This reduction leaves more Cys available for GSH synthesis under stress conditions. In addition we have also studied the effects of high GSH level on the transcriptome profile. The analysis revealed that GSH affects the expression level of many major genes coding to enzymes or proteins associated with photosynthesis, starch degradation, hormone metabolism (especially genes associated with jasmonate), biotic stress (especially genes associated with PR-proteins), cytochrome P450 genes, regulation of transcription and signaling (especially genes associated with receptor kinases and calcium). These results suggest that indeed GSH levels affect different pathways and metabolites in plants.
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McGuire, Mark A., Amichai Arieli, Israel Bruckental, and Dale E. Bauman. Increasing Mammary Protein Synthesis through Endocrine and Nutritional Signals. United States Department of Agriculture, January 2001. http://dx.doi.org/10.32747/2001.7574338.bard.
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Objectives To determine endocrine factors that regulate the partitioning of amino acids by the mammary gland. To evaluate dietary flow and supply of energy and amino acids and their effects on milk protein synthesis and endocrine status. To use primary cultures of cow mammary epithelial cells to examine the role of specific factors on the rates and pattern of milk protein synthesis. Milk protein is an increasingly valuable component of milk but little is known regarding the specific hormonal and nutritional factors controlling milk protein synthesis. The research conducted for this project has determined that milk protein synthesis has the potential to be enhanced much greater than previously believed. Increases of over 25% in milk protein percent and yield were detected in studies utilizing abomasal infusion of casein and a hyperinsulinemic-euglycemic clamp. Thus, it appears that insulin, either directly or indirectly, can elicit a substantial increase in milk protein synthesis if additional amino acids are supplied. For additional amino acids, casein provided the best response even though substantial decreases in branched chain amino acids occur when the insulin clamp is utilized. Branched chain amino acids alone are incapable of supporting the enhanced milk protein output. The mammary gland can vary both blood flow and extraction efficiency of amino acids to support protein synthesis. A mammary culture system was used to demonstrate specific endocrine effects on milk protein synthesis. Insulin-like growth factor-I when substituted for insulin was able to enhance casein and a-lactalbumin mRNA. This suggests that insulin is a indirect regulator of milk protein synthesis working through the IGF system to control mammary production of casein and a-lactalbumin. Principal component analysis determined that carbohydrate had the greatest effect on milk protein yield with protein supply only having minor effects. Work in cattle determined that the site of digestion of starch did not affect milk composition alone but the degradability of starch and protein in the rumen can interact to alter milk yield. Cows fed diets with a high degree of rumen undegradability failed to specifically enhance milk protein but produced greater milk yield with similar composition. The mammary gland has an amazing ability to produce protein of great value. Research conducted here has demonstrated the unprecedented potential of the metabolic machinery in the mammary gland. Insulin, probably signaling the mammary gland through the IGF system is a key regulator that must be combined with adequate nutrition in order for maximum response.
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Barefoot, Susan F., Bonita A. Glatz, Nathan Gollop, and Thomas A. Hughes. Bacteriocin Markers for Propionibacteria Gene Transfer Systems. United States Department of Agriculture, June 2000. http://dx.doi.org/10.32747/2000.7573993.bard.
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The antibotulinal baceriocins, propionicin PLG-1 and jenseniin G., were the first to be identified, purified and characterized for the dairy propionibaceria and are produced by Propionibacterium thoenii P127 and P. thoenii/jensenii P126, respectively. Objectives of this project were to (a) produce polyclonal antibodies for detection, comparison and monitoring of propionicin PLG-1; (b) identify, clone and characterize the propionicin PLG-1 (plg-1) and jenseniin G (jnG) genes; and (3) develop gene transfer systems for dairy propionibacteria using them as models. Polyclonal antibodies for detection, comparison and monitoring of propionicin PLG-1 were produced in rabbits. Anti-PLG-1 antiserum had high titers (256,000 to 512,000), neutralized PLG-1 activity, and detected purified PLG-1 at 0.10 mg/ml (indirect ELISA) and 0.033 mg/ml (competitive indirect ELISA). Thirty-nine of 158 strains (most P. thoenii or P. jensenii) yielded cross-reacting material; four strains of P. thoenii, including two previously unidentified bacteriocin producers, showed biological activity. Eight propionicin-negative P127 mutants produced neither ELISA response nor biological activity. Western blot analyses of supernates detected a PLG-1 band at 9.1 kDa and two additional protein bands with apparent molecular weights of 16.2 and 27.5 kDa. PLG-1 polyclonal antibodies were used for detection of jenseniin G. PLG-1 antibodies neutralized jenseniin G activity and detected a jenseniin G-sized, 3.5 kDa peptide. Preliminary immunoprecipitation of crude preparations with PLG-1 antibodies yielded three proteins including an active 3-4 kDa band. Propionicin PLG-1 antibodies were used to screen a P. jensenii/thoenii P126 genomic expression library. Complete sequencing of a cloned insert identified by PLG-1 antibodies revealed a putative response regulator, transport protein, transmembrane protein and an open reading frame (ORF) potentially encoding jenseniin G. PCR cloning of the putative plg-1 gene yielded a 1,100 bp fragment with a 355 bp ORF encoding 118 amino acids; the deduced N-terminus was similar to the known PLG-1 N-terminus. The 118 amino acid sequence deduced from the putative plg-1 gene was larger than PLG-1 possibly due to post-translational processing. The product of the putative plg-1 gene had a calculated molecular weight of 12.8 kDa, a pI of 11.7, 14 negatively charged residues (Asp+Glu) and 24 positively charged residues (Arg+Lys). The putative plg-1 gene was expressed as an inducible fusion protein with a six-histidine residue tag. Metal affinity chromatography of the fused protein yielded a homogeneous product. The fused purified protein sequence matched the deduced putative plg-1 gene sequence. The data preliminarily suggest that both the plg-1 and jnG genes have been identified and cloned. Demonstrating that antibodies can be produced for propionicin PLG-1 and that those antibodies can be used to detect, monitor and compare activity throughout growth and purification was an important step towards monitoring PLG-1 concentrations in food systems. The unexpected but fortunate cross-reactivity of PLG-1 antibodies with jenseniin G led to selective recovery of jenseniin G by immunoprecipitation. Further refinement of this separation technique could lead to powerful affinity methods for rapid, specific separation of the two bacteriocins and thus facilitate their availability for industrial or pharmaceutical uses. Preliminary identification of genes encoding the two dairy propionibacteria bacteriocins must be confirmed; further analysis will provide means for understanding how they work, for increasing their production and for manipulating the peptides to increase their target species. Further development of these systems would contribute to basic knowledge about dairy propionibacteria and has potential for improving other industrially significant characteristics.
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Sengupta-Gopalan, Champa, Shmuel Galili, and Rachel Amir. Improving Methionine Content in Transgenic Forage Legumes. United States Department of Agriculture, February 2001. http://dx.doi.org/10.32747/2001.7580671.bard.
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Leguminous forage crops are high in proteins but deficient in S- amino acids. It has been shown that both wool quality and milk production can be limited by the post-ruminal supply of sulfur-containing amino acids. Efforts to use conventional plant breeding and cell selection techniques to increase the S-amino acid content of alfalfa have met with little success. With the objective to increase the S-amino acid content of forage legumes, the goal of this project was to co- express the methionine rich zein genes from corn along with a gene for a key enzyme in methionine biosynthesis, aspartate kinase(AK). The zeins are seed storage proteins from corn and are groupec into four distinct classes based on their amino acid sequence homologies. The b-zein (15kd) and the 6zein (10kD and 18kD) have proportionately high levels of methionine (10%, 22% and 28%, respectively). Initial studies from our lab had shown that while the 15kD zein accumulated to high levels in vegetative tissues of transgenic tobacco the l0kD zein did not. However, co-expression of the 10kD zein with the 15kD zein genes in tobacco showed stabilization of the 10kD zein and the co-localization of the 10kD and 15kD zein proteins in unique ER derived protein bodies. AK is the key enzyme for producing carbon skeletons for all amino acids of the aspartate family including methionine. It is, however, regulated by end-product feedback inhibition. The specific objectives of this proposal were: i. to co-express the 15kD zein with the 10/18kD zein genes in alfalfa in order to enhance the level of accumulation of the 10/18kD zein; ii. to increase methionine pools by expressing a feedback insensitive AK gene in transformants co-expressing the 15kD and 10/18kD zein genes. The Israeli partners were successful in expressing the AK gene in alfalfa which resulted in an increase in free and bound threonine but not in methionine (Galili et al., 2000). Since our target was to increase methionine pools, we changed our second objective to replace the AK gene with the gene for cystathionine gamma synthase (CGS) in the co-expression studies. The first methionine specific reaction is catalyzed by CGS. An additional objective was to develop a transformation system for Berseem clover, and to introduce the appropriate gene constructs into it with the goal of improving their methionine content. Genes for the 15kD zein along with the genes for either the 10kD or 18kD zein have been introduced into the same alfalfa plant both by sexual crosses and by re-transformation. Analysis of these zein co-expressors have shown that both the IOkD and 18kD zein levels go up 5 to 10 fold when co-expressed with the 15kD zein (Bagga et al., MS in preparation). Incubation of the leaves of transgenic alfalfa co-expressing the 15kD and 10kD zein genes, in the rumen of cows have shown that the zein proteins are stable in the rumen. To increase the level of zein accumulation in transgenic alfalfa different promoters have been used to drive the zein genes in alfalfa and we have concluded that the CaMV 35S promoter is superior to the other strong leaf -specific promoters. By feeding callus tissue of alfalfa plants co-expressing the 15kD and 10kD zein genes with methionine and its precursors, we have shown that the zein levels could be significantly enhanced by increasing the methionine pools. We have now introduced the CGS gene (from Arabidopsis; kindly provided to us by Dr. Leustek), into the 15kD zein transformants and experiments are in progress to check if the expression of the CGS gene indeed increases the level of zein accumulation in alfalfa. We were not successful in developing a transformation protocol for Berseem clover.
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Abdo, Nabil, Dana Abed, Bachir Ayoub, and Nizar Aouad. The IMF and Lebanon: The long road ahead – An assessment of how Lebanon’s economy may be stabilized while battling a triple crisis and recovering from a deadly blast. Oxfam, October 2020. http://dx.doi.org/10.21201/2020.6652.
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Lebanon is extremely unequal and has been rocked by massive protests in recent months. The country is facing a financial crisis and is in talks with the International Monetary Fund (IMF) about a potential bailout programme. Other IMF programmes in the region have focused on austerity and have driven increases in poverty and inequality. A business-as-usual approach by the IMF in Lebanon could have serious and far-reaching adverse impacts. Any potential policies pushed by the IMF in Lebanon must first be shown not to impact negatively on economic and gender inequalities, and must be drawn up transparently in consultation with local communities, civil society organizations and social movements.
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Li, Xuan, Junjie Wang, Jing Mao, and Yunnan Li. Therapeutic effects of traditional Chinese medicine fumigating plus Yang-He decoction for patients with ankylosing spondylitis: A systematic review and network meta-analysis. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, April 2023. http://dx.doi.org/10.37766/inplasy2023.4.0074.
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Review question / Objective: To assess the therapeutic effects of combined TCMF with YHD for AS patients. Condition being studied: Traditional Chinese medicine fumigating (TCMF) and Yang-He decoction (YHD) are widely used for ankylosing spondylitis (AS), whether combined used TCMF and YHD provides superior therapeutic effects for AS remained unclear. Eligibility criteria: (1) Patients: adult patients diagnosed with AS; (2) Treatments: TCMF plus YHD, TCMF, YHD, and WM; (3) Outcomes: the primary endpoint was effective rate, and the secondary endpoints including Visual Analogue Scale (VAS) score, Schober test (ST), thoracic expansion (TE), finger-floor distance (FFD), pillow-wall distance (PWD), spinal column activity (SCA), morning stiffness time (MST), Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP); and (4) Study design: the study had to have RCT design.