Miracle-dz/UniFAR-FTUs huggingface dataset:
The 26S Proteasome and Histone Modifying Enzymes Regulate

Major Histocompatibility Complex Class-II (MHC-II) molecules are critical regulators of adaptive immunity that present extracellular antigens required to activate CD4 + T cells. MHC-II are regulated at the level of transcription by master regulator, the Class II Transactivator (CIITA), whose association with the MHC-II promoter is necessary to initiate transcription. Recently, much research focused on novel mechanisms of transcriptional regulation of critical genes like MHC-II and CIITA; findings that the macromolecular complex of the 26S-proteasome is involved in transcription have been perhaps the most exciting as they impart novel functions to a well studied system. Proteasome is a multi-subunit complex composed of a 20S-core particle capped by a 19S-regulatory particle. The 19S contains six ATPases which are required for transcription initiation and elongation. We demonstrate that 19S ATPase-S6a inducibly associates with CIITA promoters. Decreased expression of S6a negatively impacts recruitment of the transcription factors STAT-1 and IRF-1 to the CIITA due to significant loss in histone H3 and H4 acetylation. S6a is robustly recruited to CIITA coding regions, where S6a binding coordinates with that of RNA polymerase II. RNAi mediated S6a knockdown significantly diminishes recruitment of Pol II and P-TEF-b components to CIITA coding regions, indicating S6a plays important roles in transcriptional elongation. Our research is focused on the ways in which accessibility to and transcription of DNA is regulated. While cancers are frequently linked to dysregulated gene expression, contribution of epigenetics to cancers remains unknown. To achieve metastatic ability, tumors alter gene expression to escape host immunosurveilance. MHC-II and CIITA expression are significantly downregulated in highly metastatic MDA-MB-435 breast cancer cells. This suppression correlates with elevated levels of the silencing modification H3K27me3 at CIITA and a significant reduction in Pol II recruitment. We observe elevated binding of the histone methyltransferase to CIITApIV and demonstrate this enzyme is a master regulator of CIITA gene expression. EZH2 knockdown results in significant increases in CIITA and MHC-II transcript levels in metastatic cells. In sum, transcriptional regulation by the 19S-proteasome and histone modifying enzymes represents novel mechanisms of control of mammalian gene expression and present novel therapeutic targets for manipulating MHC expression in disease. INDEX WORDS: Major histocompatibility class II, Class II transactivator, Transcription regulation, 26S proteasome, 19S ATPase, S6a, epigenetic, histone modifications, histone methyltransferase EZH2 THE 26S PROTEASOME AND HISTONE MODIFYING ENZYMES REGULATE TRANSCRIPTION OF THE CLASS II TRANSACTIVATOR, CIITA
EVIDÊNCIA DA PRESENÇA DO SISTEMA AGR, EXPRESSÃO DE TOXINAS E RESISTÊNCIA AOS ANTIMICROBIANOS EM ESTAFILOCOCOS COAGULASE-NEGATIVA ISOLADOS DE HEMOCULTURAS

Coagulase–negative staphylococci (CNS) are normal resident microbes of human skin, often isolated from clinical specimens. They are the major cause of bacteremia in inpatients, especially intensive care patients. Pathogenesis of infection caused by these organisms is complex and multifactorial, with several virulence factors involved in infection processes. Biofilm is key for CNS virulence, since it provides protection against the host’s immune system and the action of antimicrobial agents. Staphylococcal toxins known as superantigens are antigenically classified as toxic shock syndrome toxin 1 (TSST-1) and enterotoxins. They trigger a chain of toxic effects by acting as potent gastrointestinal toxins and nonspecifically stimulating T cell proliferation. Staphylococcal regulatory systems, such as agr, may affect toxin and biofilm production through induction or repression of genes coding for virulence factor production specifically. In addition to virulence factors, increasing resistance rates to antimicrobial agents is also highly significant in nosocomial infections. Oxacillin–resistant strains leave few treatment alternatives for infection caused by these pathogens. Vancomycin has shown to be one drug of choice, although decreased susceptibility, as well as resistance, to vancomycin have been reported lately. Therefore, the aim of this study has been to categorize CNS species in blood culture specimens from inpatients at the UNESP Hospital das Clinicas in Botucatu, Brazil, as per detection and expression of virulence factors, agr system involved in virulence regulation, and susceptibility to antimicrobial agents.
Isolation and Characterization of a New Generalized Transducing Bacteriophage Different from Pl in Escherichia coli

A new generalized transducing bacteriophage in the Escherichia coli system was isolated and characterized. This phage, designated D108, makes clear plaques on E. coli K-10, K-12, K-12(P1kc), K-12(D6), B/r, C, and 15 T−, and Shigella dysenteriae. The plaque of phage D108 is larger in size than that of phage P1kc. Electron-microscopic observation revealed that phages D108 and P1kc are morphologically different from each other, suggesting that phage D108 belongs to a phage group different from phage P1. The fact that all of the 10 markers tested were transduced by phage D108 indicates that this phage is a generalized transducing phage in the E. coli system. The transduction frequency by phage D108 of chromosomal markers and of a drug resistance factor (R factor) ranged from 2 × 10−6 to 3 × 10−8 and 3 × 10−9 to 6 × 10−10 per phage, respectively. The cotransduction frequency of the thr and leu markers was 2.8% for phage P1kc and 1.5% for phage D108. The CM and TC markers (chloramphenicol-resistant and tetracycline-resistant markers, respectively) of the R factor were not cotransduced by phage D108, but the markers were generally cotransduced by phage P1kc. The results suggest that the transducing particle of phage D108 contains a smaller amount of host deoxyribonucleic acid than does phage P1kc.
Efeito da amamentação na reprodução de vacas: uma revisão

O longo periodo de anestro pos-parto e um dos principais fatores responsaveis pelo longo periodo de servico e consequente baixa eficiencia reprodutiva do rebanho bovino do Brasil. A amamentacao e citada como capaz de influenciar o intervalo do parto ao primeiro estro, em vacas de leite ou de corte. Esta revisao mostra varios resultados de pesquisa sobre os efeitos da amamentacao na reproducao, e descreve as inumeras vias pelas quais a succao do ubere pelo bezerro poderia afetar a atividade ovariana. O objetivo principal e fornecer uma base para melhor entendimento e maiores investigacoes dos mecanismos especificos pelos quais a funcao reprodutiva e afetada pela amamentacao.
The Laboratory Fish

Part 1: Introduction(Diversity of fish, early observations and descriptions, fish in experimentation) Part 2: Housing, Maintenance, and Breeding Facilities and Husbandry (Large Fish Models) Facilities and Husbandry (Small Fish Models) Diet Common Diseases and Treatment Part 3: Gross Functional Anatomy Integumentary System Skeletal System Muscular System Nervous System Respiratory System Circulatory System Digestive System Urinary Tract Endocrine System Immune System Sensory Systems Reproductive Systems Part 4: Microscopic Functional Anatomy Integumentary System Skeletal System Fish as an Experimental Model for Studying Muscule Function Nervous System Respiratory System Circulatory System Digestive System Urinary Tract Endocrine System Immune System Sensory Systems Reproductive Systems Part 5: Procedures Stress and Anesthesia Collection of Body Fluids Routes of Administration for Chemical Agents Fish Necropsy Surgical Techniques Fixation of Fish Tissues Autoradiography of Fishes Part 6: Experimental Models Cancer Toxicology Cell and Tissue Culture Glossary Index
Effect of Field Dodder (Cuscuta campestris Yuncker) on Some Legume Crops

Twelve legume crops were tested to study the effect of field dodder (Cuscuta campestris Yuncker) on them. The tested crops showed great variations in response to field dodder parasitism. Based on the reduction in host dry weight (biological yield) caused by the parasite, the tested crops were classified into three groups: crops that lost > 50% of the their biological yield were considered highly susceptible, those that lost 10-50% of their biological yield were considered susceptible and those that lost < 10% of their biological yield were considered resistant. The results of the study revealed that among the tested legume crops the first group (the highly susceptible) comprised 6 crops (hyacinth bean, lens, chickpea, broad bean, alfalfa, fodder pea), the second group (the susceptible) comprised 4 crops (fenugreek, Egyptian clover, lupin, garden pea) and the third group (the resistant) comprised 2 crops (kidney bean and vetch). The parasitic damage appeared to be of greater magnitude on hyacinth bean (75% loss of biological yield) and of lesser magnitude on kidney bean (8.1% loss of biological yield) as compared to the other legumes. However, in vetch the parasite had a positive effect on its growth traits (17% increase in biological yield). The significant reductions in the growth parameters of susceptible and highly susceptible crops were associated with a marked increase in shoot yield of field dodder.
Home range size and spatial distributions of two sympatric lizard species (Sceloporus undulatus, Holbrookia maculata) in the Sand Hills of Nebraska.

Home range size and spatial distributions were determined for sympatric populations of the prairie lizard (Sceloporus undulatus) and the lesser earless lizard (Holbrookia maculata) in the Sand Hills of Nebraska. Mean home ranges of 717 ml and 852 m2 were found for males and females, respectively, of S. undulatus. Male H. maculata had a mean home range of 3822 m2 whereas females had a mean home range of 1823 M2. The two species tended to occupy different microhabitats. The association of S. undulatus with structural features of the habitat such as clumps of Yucca accounted for the similarity in home ranges of the two sexes. There was also evidence of territoriality and social relationships by male Sceloporus. The expanded home ranges of male H. maculata may be an adaptation to increase the probability of encountering females.
Role of Epigenetic Modifications and Dexras1 in glucocorticoid regulation of growth hormone expression

Title of Document: ROLE OF EPIGENETIC MODIFICATIONS AND DEXRAS1 IN GLUCOCORTICOID REGULATION OF GROWTH HORMONE EXPRESSION Jyoti Narayana Doctor of Philosophy, 2011 Directed By: Professor Tom E. Porter, PhD Department of Animal and Avian Sciences Biological Sciences Graduate Program Glucocorticoid induction of growth hormone in anterior pituitary cells of embryonic chickens is an indirect effect. A genome-wide microarray screen conducted on embryonic day 11 chickens identified Dexras1 as a novel candidate gene involved in mediating the glucocorticoid effects on GH mRNA expression. The following studies were aimed at characterizing glucocorticoid regulation of chicken Dexras1 and identifying potential involvement of Dexras1 in mediating glucocorticoid effects on GH expression. We determined that glucocorticoid induction of Dexras1 requires glucocorticoid receptor and both MEK1/2 and Ras cell signalling pathways. Overexpression of Dexras1 vector had no effect on GH reporter activity. Transfection of 10ng or 100ng of Dexras1 expression plasmid stimulated basal GH mRNA levels, whereas transfection of 1000ng of Dexras1 expression plasmid significantly inhibited levels of glucocorticoid-induced GH mRNA. YFP-tagged Dexras1 protein was predominantly found in the cytoplasm and glucocorticoid treatment did not cause Dexras1 to translocate to the plasma membrane. Downstream targets of Dexras1 are not known. Additionally, glucocorticoid-regulated histone modifications within 2kb of the GH gene were investigated using chromatin immunoprecipitation assays. The effects of glucocorticoid (1.5 h or 6 h) on histone H3 modifications, RNA Polymerase II (Pol II) recruitment, and association of GR, Pit-1, and Ets1 with the chicken GH gene were examined. We found increased H3 acetylation and tri-methylation of lysine 4 at both Pit1 sites and the transcription start site (TSS) in response to 1.5 h glucocorticoid treatment. Furthermore, 1.5 h glucocorticoid treatment significantly increased recruitment of Pit-1 and Pol II to the proximal Pit-1 site and the TSS, respectively. GR was recruited to the glucocorticoid-responsive region (GCRR) and the distal Pit-1 site in response to 1.5 h glucocorticoid treatment, while 6 h glucocorticoid treatment resulted in Ets1 dissociation from the GCRR. Collectively, these results indicate that glucocorticoid induces dynamic changes in histone modifications and transcription factor recruitment within the 5‘flanking region of the chicken GH gene. We conclude that glucocorticoid induction of GH gene expression in chickens is achieved by a combination of genomic and nongenomic pathways. Our studies provide novel mechanisms of GH regulation in chickens, some of which may be found in other vertebrate species as well. ROLE OF EPIGENETIC MODIFICATIONS AND DEXRAS1 IN GLUCOCORTICOID REGULATION OF GROWTH HORMONE EXPRESSION
A Primary Investigation on the Culture-dependent Moderately Halophilic Bacteria Colonizing in Dagong Ancient Brine Well, Sichuan

aerobic moderate halophilic bacteria were isolated from Daong Ancient Brine Well located in Zigong city, Sichuan Province, China. And they were subjected to the analysis of phenotype, physiology, 16S rRNA sequence. Furthermore, the intra specific phylogeny of closely related strains was also screened by PCR fragment length polymorphism of 16S-23S ribosomal RNA intergenic spacer regions (ISR). The re- sult showed that the halophilic isolates found in current study were closely related to the following genera:
Parasitas intestinais em suínos confinados nos Estados de São Paulo e Minas Gerais

The presence of intestinal parasites in confined swineherds from states of Sao Paulo and Minas Gerais was evaluated by means of faecal exams. The installations used at the growing and fattening phases differ among the farms by the presence or not of water slides. Eimeria spp. , Isospora suis, Cryptosporidium spp. , Balantidium coli, Strongyloidea and Ascaris suum were identified in piglet faeces. Sows were infected by Eimeria spp. , Balantidium coli, Strongyloidea and Ascaris suum. Association, measured by Fisher exact test, showed that the presence of Strongyloidea helminths was associated with the management. The management and routine use of ivermectin and fenbendazole improve the herd sanitary condition, but parasite infections still persist. The main source of infection in pigs raised in confinement is the environment. Resistance features of some parasite eggs and oocysts to environmental conditions, mainly the humidity, are important factors to the occurrence of the infection on these farms.
Protein aggregation, oxidative stress and the role of the yeast peroxiredoxin Tsa1.

Peroxiredoxins are ubiquitous, thiol-specific proteins that have multiple functions in stress protection, including oxidative stress. Tsa1 is the major yeast peroxiredoxin and we show that it functions as a specific antioxidant to protect against oxidative stress caused by nascent protein misfolding and aggregation. Yeast mutants lacking TSA1 are sensitive to misfolding caused by exposure to the proline analogue azetidine-2-carboxylic acid (AZC). AZC promotes protein aggregation and its toxicity to a tsa1 mutant is caused by reactive oxygen species (ROS). Generation of [rho0] cells lacking mitochondrial DNA rescues the tsa1 mutant AZC sensitivity indicating that mitochondria are the source of ROS. Inhibition of nascent protein synthesis with cycloheximide prevents AZC-induced protein aggregation and abrogates ROS generation confirming that aggregate formation causes ROS production. Protein aggregation is accompanied by mitochondrial fragmentation and we show that Tsa1 localizes to the sites of protein aggregation, which are formed adjacent to mitochondria.Further investigation reveals that AZC-induced protein aggregation leads to an inhibition of mitochondrial respiration and the depolarisation of the mitochondrial membrane. Remarkably, this was entirely dependent on the presence of Tsa1. We show that the effects of protein aggregation on mitochondrial function are mediated by the Ras/PKA pathway and that Tsa1 appears to influence the activity of this pathway through its effects on the yeast phosphodiesterase, Pde2. Together, these data indicate a new role for peroxiredoxins in the response to ROS, generated as a result of protein misfolding and aggregate formation. Finally, we analysed the characteristics of proteins found within protein aggregates, isolated from different conditions during the course of the study. Our results highlight the differences between proteins that aggregate under normal, mid-exponential growth conditions (physiological aggregates) and those which aggregate during cellular stress. We were able to establish the characteristics of an archetypical physiological aggregate, through an assessment of a range of properties, identifying factors that significantly differed from genomic expectations. Furthermore, our observations indicate that, in general, cellular stress reduces the threshold of metrics associated with protein aggregation propensity. We also found that different stresses result in the aggregation of proteins that are, largely, physicochemically indistinct from one another, regardless of the mode of toxicity. Finally we show that a significant number of proteins, identified in our protein aggregates, were also present in protein aggregates isolated from aged C. elegans. This suggests that the factors and components of protein aggregates are conserved.
Inhibition of Apoptosis is Responsible for the Acquired Resistance of K562 Cells to Cisplatin

In all attempt to elucidate the role of apoptosis in drug resistance, cisplatin-resistant human chronic myelogenous leukemia (CML) K562 cells (K562/CDDP) were established and compared with drug sensitive parent cells (K562) in the induction of apoptosis. K562/CDDP cells were 5-fold more resistant to cisplatin compared to K562 cells. In addition, K562/CDDP cells were significantly more resistant to apoptois as judged by DNA fragmentation and DAPI staining. K562/CDDP cells exhibited decreased proleolytic activity of caspase-3 and this was further demonstrated by decreased cleavage of its substrate poly (ADP-ribose) polymerase (PARR- Western blot analysis showed that K562/CDDP cells had longer sustained levels of BCL- whereas no difference was noted in the level of Bcl-2. the translocation of Bax to mitochondria was significantly delayed in K562/CDDP cells. These results suggest that the reduced translocation of Bax and the sustained expression of BCL- may cause resistance to apoptosis through prevention of mitochondria release of cytochrome c, which subsequently induces reduction of caspase-3 activity and that this response is partly responsible for the acquired resistance to cisplatin ill K562 cells.
Identification and Functional Characterisation of the Novel OCRL1 Binding Partners IPIP27A and B

Mutation of the inositol polyphosphate 5-phosphatase OCRL1 results in two disorders in humans, namely Lowe syndrome, characterized by ocular, nervous system and renal defects, and Type 2 Dent? s disease, in which only the renal symptoms are evident. The disease mechanisms of these syndromes are poorly understood. In an effort to better understand how Lowe syndrome is caused we have identified two novel OCRL1-binding proteins, termed inositol polyphosphate phosphatase interacting protein of 27kDa (IPIP27) A and B. IPIP27A and B bind to the C-terminal region of OCRL1 via a conserved motif that also binds the signaling adaptor protein APPL1. Both IPIP27A and B localise to early and recycling endosomes and the trans-Golgi network (TGN). We have studied the function of these proteins using a variety of techniques, including overexpression of wild type and mutant IPIP27 constructs and RNAi mediated depletion of endogenous IPIP27A/B in cell-based functional assays. Our studies show that IPIP27A/B are required for efficient receptor recycling from endosomes to the TGN and plasma membrane. In support of this hypothesis, we have now identified several IPIP27A interaction partners including the F-BAR domain protein PACSIN2, which is thought to deform membranes during formation of trafficking intermediates. Our results identify IPIP27A and B as key players in endocytic trafficking, and suggest that they function as scaffolding proteins required for the formation of transport carriers from endosomal and Golgi membranes. Together our results strongly suggest that defects in this process are responsible for pathology of Lowe syndrome and Dent? s disease.
UJI BANDING EFEKTIVITAS VIRGIN COCONUT OIL DENGAN KETOKONAZOL 2% SECARA IN VITRO TERHADAP PERTUMBUHAN Candida albicans

Background : Candida albicans, an extremely widespread yeast, is the major cause of candidiasis. Virgin coconut oil consist of fatty acids which were shown to have antimicrobial activity against Candida albicans. Ketoconazole inhibits hepatic cytochrome P450 isozymes so that ergosterol synthesis is blocked and fungal cell membrane damage is occurred. The aim of the study is to investigate the possible antimicrobial activity of virgin coconut oil against Candida albicans by comparing them with the antimicrobial activity of 2% ketoconazole. Method : The study is a laboratory experimental design. The samples consist of 31 Candida albicans isolates. One C.albicans strain is obtained from the American Type Culture Collection (ATCC 10231), the others are recent clinical isolates which is isolated from cervical swab and identified by germ tube test. The antimicrobial activity is determined with the disc diffusion method. Result : Research was resulted that virgin coconut oil can not inhibit the growth of Candida albicans. Conclusion : Virgin coconut oil is not effective in inhibitting the growth of Candida albicans in vitro. Keyword : Candida albicans, virgin coconut oil, ketoconazole