Miracle-dz/UniFAR-FTUs huggingface dataset:
The Relationship of Galectin-1 and Neuroblastoma Stem Cells

Introduction : Neuroblastoma is the most common extracranial solid tumor of childhood. It originates from the primordial neural crest cells. The cancer stem cell (CSC) are a small population of tumor cells, termed CSCs that have the ability to proliferate and maintain tumor growth. CSCs have two main characteristics: self- renewal and differentiation. Galectin-1 (Gal-1) has important roles in the cell adhesion and growth, apoptosis, immunomodulation, metastasis, inflammation and mRNA splicing. In this study, we aimed to investigate the expression of Gal-1 in the CD133+ stem cells of two neuroblastoma cell lines with different prognostic characteristics in comparison with their CD133- cells. Materials and Method : We cultured neuroblastoma cell lines SH-SY5Y (N-myc negative) and Kelly (N-myc positive) in DMEM and RPMI 1640 mediums and cultivated in an incubator equipped with 5% CO2 at 37Â°C. We isolated CD133+ neuroblastoma stem cells with magnetic beads. We spreaded the isolated CD133+ and CD133- Kelly and SHSY5Y neuroblastoma cells onto the polylysine-coated slides and stained with Gal-1 immunocytochemically. We counted 50 different 100 cells for each group and evaluated Gal-1 expression as positive (middle and high expression) and negative (weak and no expression). The results were statistically analyzed with the Mann-Whitney U non-parametric test. Results : The control cells had included both of the CD133+ and CD133- cells. When we evaluated Gal-1 expressions in control cells, Kelly control cells showed more Gal-1 expression than SHSY5Y control cells and this higher expression was more obvious in CD133+ Kelly stem cells. Gal-1 expression in CD133- cells (other cells) was very low. When we evaluated Gal-1 expression in CD133+ SHSY5Y stem cells and in CD133- other SHSY5Y cells, we did not determine a significant difference between them.
Wildlife Conservation Outside Protected Areas—Lessons from an Experiment in Zambia

: Managing an area as a national park in Africa often entails dealing with human pressures from regions surrounding the protected area Dealing with these human pressures is the subject of this paper, which goes well beyond mere law enforcement to discuss the realm of traditional leadership in the village, rural socioeconomics, and the services of wildlife management by government in areas surrounding a national park. This paper describes the results of an experimental project in Zambia that sought to halt the drastic loss of elephants and rhinos to poaching in and around protected areas in the Luangwa Valley. The design of this project was to involve local residents outside national parks in wildlife protection and management activities. In addition, sustained-yield uses of wildlife were adopted to recycle revenue shares into community development and to make the program of wildlife management financially self-supporting. To help carry out the various tasks of protecting and developing wildlife resources, residents were trained and employed as wildlife management staff Village wildlife councils provided advice and assistance. Safari concessions yielded sufficient revenue to meet the recurrent costs of wildlife management and also generated significant earnings for local community benefits. As a result; poaching dropped dramatically, local economies were improve and village attitudes toward wildlife management and conservation became more positive.
EL REINO PLANTAE: ¿QUÉ ES UNA PLANTA Y CÓMO SE CLASIFICAN?

Hoy en día existe un cambio de paradigmas en la Botánica, que es fundamental analizar y debatir. Los principales conceptos que están bajo una intensa discusión y cambios son la filogenia y clasificación biológica de los seresvivos con cloroplastos, la nueva visión del concepto de “planta” y “célulavegetal” y el reemplazo de la teoría celular por una visión organísmica de las plantas multicelulares, lo que ha generado una visión holística de las mismas. Los objetivos del presente trabajo son presentar y debatir la actual circunscripción de los conceptos de “planta” y “célula vegetal” y presentar los recientes avances en el campo de la filogenia y clasificación de las plantas.
Environmental pollution and its impact on domestic animals and wildlife

Environmental pollution is a major global problem posing serious risk to man and animals. It is defined as the human alteration of chemical or physical characteristics of the environment to a degree that is harmful to living organisms. Rapid pace of urbanization, industrialization and indiscriminate and improper use of chemicals such as pesticides and drugs have resulted in increased contamination and degradation of the environment leading to adverse health effects on living beings and problems associated with residues in food of animal origin. Recent emergences in environmental related toxicity in man and animals and detection of chemical residues in food products at an increasing frequency have underlined the impact of pollution on biotic component. The deleterious health effects may be observed in the form of overt clinical signs and higher morbidity and mortality or as subtle or subclinical effects. Pesticide toxicity, plumbism, mercury poisoning and fluorosis are some of the commonly encountered overt toxic problems due to pollution in domestic animals. The low level of exposure to pollutants is often associated with chemical residues in animal system, and subtle or subclinical effects. These include oxidative stress, immunotoxicity, carcinogenicity, teratogenecity and endocrine disruption. Wild animals, like their domestic counterparts also suffer from adverse effects of pollution. Eggshell thinning in birds of prey due to DDT residues and heavy mortality amongst fish and birds due to contamination of aquatic ecosystem with mercury, oil spills or acid rains are some of the notable examples of impact of pollution on wild life. Recently, decline in vulture population in Indian subcontinent has been linked to drug residues. This has drawn the attention of researchers and environmentalist owing to drastic decline in the population of an important raptorial bird species and resultant ecological degradation. Besides deleterious health effects on animal and wildlife, chemical pollutants also pose public health concerns because of contamination of human food chain. Noise and other physical pollutants are also reported to cause adverse effects on health and physiology. This review describes the incidence, source of pollutants, adverse health effects and scientific achievements in the field of environmental pollution with particular reference to animal and wildlife health.
Structural Requirements for Ribosome-Dependent GTPase Activity and Binding

iv Acknowledgements . vii List of Figures . xi Table of Abbreviations . xiv Chapter 1 – Ribosomes: Form and Function. 1 Ribosome Structure . 2 tRNAs . 5 Translation . 7 Ribosomal Protein L7/L12 . 18 Chapter 2: GTPases . 23 GTPases and GTPase Superfamily . 23 Homology of translational GTPases . 25 Translational GTPase activation . 29 Elongation Factor G . 30 GTPase-70S binding regions . 34 Project Goals . 37 ix Chapter 3 – Materials and Methods . 38 Buffers . 38 Generation of EF-G Domain Mutants . 40 Transformation of expression vectors into Escherichia coli . 41 Restriction Digestion . 42 Overexpression of (His)6-tagged translation factors . 43 Purification of (His)6-tagged translation factors . 44 Electrophoresis . 44 Anion exchange chromatography . 45 Size exclusion chromatography . 46 Purification of (His)6-tagged 70S ribosomes . 46 Purification of untagged 70S ribosomes . 47 Purification of L12 . 49 Depletion and reconstitution of L12 . 49 Western blots . 50 Malachite green GTPase activity assay . 51 Removal of (His)6-tag from GTPases . 52 x GTPase binding assay through ultracentrifugation . 52 Quantification of SDS-PAGE gels using ImageJ . 53 BioLayer Interferometry Kinetics assay . 53 Circular Dichroism . 54 Chapter 4 – Role of L12 in translational GTPase Activation and binding . 55 Results . 55 Discussion. 73 Conclusions. 82 Future Work. 83 Chapter 5 – Effect of EF-G domain deletion on GTP hydrolysis Activity . 84 Results . 84 Discussion. 97 Conclusions. 101 Future Work. 101 References . 103
Glycine-Containing Axon Terminals on Large Ventral Horn Cells Including Identified α-Motoneurons in the Spinal Cord of the Chicken

Glycine-like immunoreactive (LI) axon terminals were observed on large ventral horn cells by the indirect antibody peroxidaseantiperoxidase technique in the chicken. The glycine-LI terminals were also confirmed on identified a-motoneurons by retrograde transport of horseradish peroxidase (HRP). Electron microscopically, glycine-LI terminals containing pleomorphic clear vesicles made axosomatic and axodendritic synapses with large ventral horn cells including identified a-motoneurons. They formed mostly symmetrical synapses, but a few asymmetrical synapses were also seen. These glycine-LI terminals contained numerous mitochondria. The number of boutons per perimeter of a large ventral horn cell was 0.15±0.05/μm (n=90). The results indicate that α-motoneurons receive glycinergic inputs. The origin of glycinergic inputs is discussed in the light of previous investigations.
Arresting Features of Bacterial Toxins

Bacteria produce an arsenal of sophisticated toxins that disrupt the normal processes of the host cell, usually by modifying or inactivating host cell proteins. Now, as Coburn and Leong discuss in their Perspective, members of the cytolethal distending toxin (CDT) family have been identified as enzymes that attack DNA (and not protein) within the host cell (Lara-Tejero and GalAin). By attacking DNA, perhaps during chromosomal replication, CDTs cause the host cell to halt in Gphase of the cell cycle.
Nucleic acid specific Toll-like receptors in lupus nephritis

Nucleic acids that occur free or as immune complexes may trigger immune activation leading to aggravation of diseases with autoimmune predisposition. TLR3 and TLR 9 represent receptors that signal for viral and bacterial nucleic acids respectively. pI:C RNA, a synthetic double stranded RNA with identical properties to that viral origin activates TLR3 led to aggravation of lupus nephritis, a form of immune complex glomerulonephritis, in pre-existing lupus in MRLlpr/lpr mice. Exposure to pI:C RNA (a structural analogue of viral dsRNA) can aggravate lupus nephritis through TLR3 on antigen-presenting cells and glomerular mesangial cells. pI:C RNA–induced cytokine and chemokine production represents a major mechanism in this context. Likewise, CpG DNA a classical activator for TLR-9 led to disease aggravation in this mouse model albeit, through mechanisms that shared some commonality as well as differences to that observed with pI:C RNA. Apparently, pathogen associated immunomodulation relates to the cell-type-specific expression pattern of the respective pattern-recognition receptor. dsRNA-induced disease activity is independent of B cell activation and humoral antichromatin immunity in experimental SLE and therefore differs from CpG-DNA–induced autoimmunity. These findings contribute to the understanding of pathogen-associated modulation of autoimmunity but may also be involved in the pathogenesis of other types of inflammatory kidney diseases, e.g., flares of IgA nephropathy, renal manifestations of chronic hepatitis C virus infection, and renal vasculitis. Further, besides signifying the role of foreign and self-DNA as a pathogenic factor in autoimmune disease activity in lupus, this detailed study reveals, that certain synthetic G-rich nucleic acids may potentially block nucleic acid specific TLR functions and thus prove beneficial in arresting disease activity during progressive systemic lupus. One such G-rich DNA employed in this study has proven to be beneficial and suppressed systemic lupus in MRLlpr/lpr mouse model. Thus, modulating the CpG-DNA - TLR9 pathway may offer new opportunities for the understanding and treatment of lupus.
Degradation of ribulose‐1,5‐bisphosphate carboxylase and chlorophyll in senescing barley leaf segments triggered by jasmonic acid methylester, and counteraction by cytokinin

Barley (Hordeum vulgare L. cv. Salome) primary leaf segments responded to the application of a putative plant growth regulator, ± jasmonic acid methylester (JA-Me). with accelerated senescence, as indicated by the loss of chlorophyll and the rapid decrease in activity and immunoreactive protein content of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBP carboxylase, EC 4.1.1.39). The senescence-promoting action of JA-Me differed in light and in darkness; e.g. the initial rates of chlorophyll and RuBP carboxylase breakdown were markedly higher in light than in darkness in the presence of 4.10−5M JA-Me. Cytokinin (benzyladenine, 4.10−5M) stopped the loss of chlorophyll and RuBP carboxylase during senescence; however, the rapid drop induced by JA-Me in the early phase of leaf segment senescence could not be prevented by concomitant or previous addition of BA. On the other hand, BA added 24 h after JA-Me application resulted in a recovery of chlorophyll and RuBP carboxylase at the later stages, indicating a possible rapid inactivation of JA-Me in the tissues. The activities of a number of other chloroplastic and cytosolic enzymes were not significantly altered in JA-Me-treated leaf segments compared with controls floated on water. Time-dependent chlorophyll decrease in isolated chloroplasts did not change upon JA-Me addition to the isolated organelles. It is suggested that JA-Me acts on chloroplast senescence by promoting cytoplasic events which eventually bring about the degradation of chloroplast constituents.
Monitoring the Hemolytic Effect of Mesoporous Silica Nanoparticles after Human Blood Protein Corona Formation

The interaction of promising nanoparticles with red blood cells (RBCs) is a critical point to be addressed in nanomedicine and nanotoxicology, and the hemolytic assay is a classical and common test used to evaluate such interactions and the consequent nanoparticle toxicity. In addition, the protein corona is an emergent concept in bionanoscience associated with the manifestation of energetically driven protein–nanoparticle interactions, with a great impact on the nanomaterial toxicity assessment. In the convergence of these two concepts, we evaluated the influence of the formation of the protein corona during the hemolysis induced by spherical mesoporous silica nanoparticles with silanol groups on the external surface (MSN-SiOH), which present a confirmed toxicity on RBCs when they are dispersed as a colloid in phosphate buffer saline solution (PBS). It was observed that human blood proteins such as human serum albumin (HSA), human plasma (HP), hemoglobin (Hb), and RBC lysate, termed hemolysate (HL), can suppress the hemolytic effect induced by MSN-SiOH in a dose-dependent manner. The EC50 values of hemolysis suppression were 24, 8.0, 19, and 28 μg mL–1 for HSA, HP, Hb, and HL, respectively. This work thus shows that the results of the hemolytic assay that defines the toxicity and bioreactivity of silica nanoparticles (and others) must be interpreted as a function of the formation of the protein corona.
Prokaryotic Arsenic Resistance - Studies in Bacillus subtilis

Arsenic is a toxic metalloid which is found all over the globe. Due to its toxicity and wide abundance, all living organisms have evolved intrinsic arsenic resistance systems. In this study, three proteins that provide arsenic resistance in the Gram-positive bacterium Bacillus subtilis have been investigated. Acr3 is a trans-membrane protein that extrudes arsenite to the cell exterior. It belongs to the ACR family of arsenite transporters. Experimental data on the topology of Acr3, the first ever for a member of the ACR family, is provided and show that Acr3 has 10 trans-membrane helices. Both the N- and C-terminal ends of Acr3 are located to the cytoplasm and the protein has unusually short loops connecting its helices. ArsR is an arsenite sensitive transcription regulator that controls the expression of genes encoding arsenic resistance proteins. Experiments show that the operator site for ArsR from B. subtilis consists of a 6-6-6 inverted repeat and that DNA binding by ArsR involves formation of higher order multimers of the protein. The ArsK (former YqcK) protein has an unknown function. The present study shows that the arsK gene contributes to resistance towards both arsenite and arsenate. The results indicate that the function of ArsK is important in an aerobic environment and that it decreases the inhibitory effect that arsenite has on the sporulation efficiency of B. subtilis. A function of ArsK that involves an enzymatic addition of low molecular weight thiols to arsenic is proposed.
Influence of protein concentration on farinograph absorption, mixing requirements and mixing tolerance

Accurate and precise measurements of grain quality traits are required throughout the wheat (Triticum aestivum L. Em. Thell.) production and marketing chain to ensure that quality standards are maintained. In the present study, spring and winter wheat genotypes representing a wide range of grain quality types and protein concentration (PC) were grown in dry land and irrigated nitrogen fertilizer trials in western Canada to determine the influence of flour PC and quality on farinograph water absorption, mixing requirements and mixing tolerance. As expected, there was a strong linear relationship between PC and water absorption. In contrast, PC had a large nonlinear influence on mixing requirements and tolerance measurements, which made it difficult to make meaningful comparisons among genotypes when flour PC was less than 12%. Consequently, because cultivar grain yield potential, environment, and fertilizer management decisions have a large and variable influence on PC, it is important that cultivar select.
Analysis of G Protein βγ Dimer Formation in Live Cells Using Multicolor Bimolecular Fluorescence Complementation Demonstrates Preferences of β1 for Particular γ Subunits

The specificity of G protein βγ signaling demonstrated by in vivo knockouts is greater than expected based on in vitro assays of βγ function. In this study, we investigated the basis for this discrepancy by comparing the abilities of seven β1γ complexes containing γ1, γ2, γ5, γ7, γ10, γ11, or γ12 to interact with αs and of these γ subunits to compete for interaction with β1 in live human embryonic kidney (HEK) 293 cells. βγ complexes were imaged using bimolecular fluorescence complementation, in which fluorescence is produced by two nonfluorescent fragments (N and C) of cyan fluorescent protein (CFP) or yellow fluorescent protein (YFP) when brought together by proteins fused to each fragment. Plasma membrane targeting of αs-CFP varied inversely with its expression level, and the abilities of YFP-N-β1YFP-C-γ complexes to increase this targeting varied by 2-fold or less. However, there were larger differences in the abilities of the CFP-N-γ subunits to compete for association with CFP-C-β1. When the intensities of coexpressed CFP-C-β1CFP-N-γ (cyan) and CFP-C-β1YFP-N-γ2 (yellow) complexes were compared under conditions in which CFP-C-β1 was limiting, the CFP-N-γ subunits exhibited a 4.5-fold range in their abilities to compete with YFP-N-γ2 for association with CFP-C-β1. CFP-N-γ12 and CFP-N-γ1 were the strongest and weakest competitors, respectively. Taken together with previous demonstrations of a role for βγ in the specificity of receptor signaling, these results suggest that differences in the association preferences of coexpressed β and γ subunits for each other can determine which complexes predominate and participate in signaling pathways in intact cells.
Role of short chain fatty acid receptors in the gastrointestinal tract and their potential involvement in appetite control

The objective was to explore the role of GPR41 (FFAR3) and GPR43 (FFAR2) in colonic physiology and potential involvement in appetite control. The expression of SCFA receptors mRNA across the longitudinal axis of healthy mice, pig and human colon were found to be similar. Immunohistochemistry revealed FFAR2 and FFAR3 to be localised to flask shaped cells deep in colonic crypts and were identified as open type enteroendocrine cells by co-localisation with chromogranin A, a classical marker of enteroendocrine cells using both pig and human colon tissue. Further co-localisation studies with pig colonic tissue showed the SCFA receptors are co-localised with serotonin (enterochromaffin cells), and the satiety peptides, peptide YY (PYY) and glucagon like peptide 1 (GLP-1), indicative of enteroendocrine L cells. Impact of dietary fibre was assessed using colonic tissue collected from pigs fed a fermentable carbohydrate diet were compared to a control diet of less fermentable carbohydrate. Results demonstrate steady state SCFA concentrations exist in the colonic lumen, explained by enhanced uptake of SCFA due to an increased expression of SCFA transporter (MCT1). Dietary fibre was found to have minimal impact on relative abundance of the SCFA receptors mRNA across the longitudinal axis of pig colon. However, evaluation of the satiety peptides, PYY and pro-glucagon (precursor of GLP-1) mRNA have revealed a statistical significant increase in PYY but not pro-glucagon mRNA. Subsequently, attention was focused toward use of in-vitro models to explore SCFA receptors in appetite control. Initial in-vitro experiments assessed activity profile of SCFA receptors and evaluated chemical tools used to differentiate between FFAR2 and FFAR3. Murine ‘GLUTag’ and human ‘NCI-H716’ in-vitro models of enteroendocrine ‘L cells’ were characterised and SCFA induced GLP-1 secretion evaluated. 10mM butyrate and 10μM 4-CMTB induced GLP-1 release. RNA interference was used in attempt to knockdown FFAR2 in NCI-H716 cells to evaluate SCFA sensor’s involvement in GLP-1 release. In summary, luminal SCFA may stimulate SCFA receptors of enteroendocrine cells releasing peptides; GLP-1, PYY or serotonin to co-ordinate gut motility and appetite.
A Simple Model to Deal with Sire by Treatment Interactions When Sires Are Related

Abstract The assumption that var ( hs ) = I σ hs 2 is not valid when sires are related. A more logical assumption is that the variance is I ⊗ A σ hs 2 . This result is derived from the use of an equivalent multiple-trait model. This procedure was extended to a more complicated model involving treatments and random herds within treatments. A small simulation trial demonstrated that σ hs 2 is underestimated if the I σ hs 2 model is used when the true model should be I ⊗ A σ hs 2 .