Miracle-dz/UniFAR-FTUs huggingface dataset:
Structural Characterization And Therapeutic Utility Of The Proton-Coupled Folate Transporter

STRUCTURAL CHARACTERIZATION AND THERAPEUTIC UTILITY OF THE PROTON-COUPLED FOLATE TRANSPORTER by MICHAEL ROY WILSON December 2016 Advisor: Dr. Larry H. Matherly Major: Cancer Biology Degree: Doctor of Philosophy Folate is a B9 vitamin essential to DNA synthesis. The proton-coupled folate transporter (PCFT) is a newly discovered proton/folate symporter with an acidic pH optimum and broad expression across a variety of solid tumor types, with limited expression in normal tissues. Several antifolate molecules have been developed as cancer therapeutics, although these classical antifolates display numerous off-target effects due to transport by the ubiquitous reduced folate carrier (RFC). In this dissertation, we determine the roles of multiple PCFT structure/function domains, and develop PCFT-specific antifolates to target solid tumors. We utilize substituted cysteine accessibility methods (SCAM) to identify a novel reentrant loop structure between the second and third transmembrane domains (TMDs), which is critical to PCFT function, using membrane-impermeable thiol-reactive reagent MTSEA-biotin. We also utilize SCAM to identify and characterize the PCFT oligomerization interface at TMDs 3 and 6 via crosslinking reagent MTS-6-MTS. By the development of PCFT and ThTr1 chimera molecules, creation of PCFT half molecules, and cysteineand alanine-scanning mutagenesis, we determined the purely structural role of the TMD6-7 connecting loop region. We studied novel 6-substituted pyrrolo[2,3-d]pyrimidine thienoyl regioisomers related to pemetrexed (PMX), a standard NS-
Obtención, almacenamiento y morfometría de espermatozoides aviares: aplicación para la caracterización y criopreservación de espermatozoides de especies silvestres

Esta tesis ha sido pensada y estructurada para estudiar los aspectos basicos de la espermatologia aviar y desarrollar las tecnicas de criopreservacion de semen en diferentes especies de aves. Estas tecnicas, que forman parte de las denominadas tecnicas de reproduccion asistida o TRA estan ganando cada vez mas protagonismo en la mejora y preservacion tanto de razas como de especies. El desarrollo de estas tecnicas ha permitido la proliferacion de proyectos de cria en cautividad apoyando los programas de conservacion de especies autoctonas y han supuesto una importante mejora en el manejo genetico de las poblaciones. Aun asi, a pesar del exito obtenido con estas tecnicas en algunas especies de aves, son tecnicas apenas desarrolladas en aves silvestres, en gran parte debido a la falta de conocimientos sobre los aspectos fisiologicos y las caracteristicas bilogicas de los espermatozoides de estas especies. El estudio de los aspectos biofisicos de los gametos y del tejido gonadal de las diferentes especies es el metodo mas efectivo para desarrollar protocolos apropiados de criopreservacion. Ademas en las diferentes especies de aves silvestres el acceso al material genetico y las muestras viables de semen es dificil, consecuencia de su anatomia y fisiologia, y muy reducida en el tiempo, debido a la breve estacion reproductiva que presentan y por lo tanto las tecnicas de criopreservacion representan, ademas, una posibilidad de mejorar el aprovechamiento de los recursos disponibles. Es por ello que en el trabajo se planteo en primer lugar el desarrollo y evaluacion de las tecnicas de recoleccion post-mortem de semen, empleando como modelo una especie de ave domestica, el gallo (Capitulo 1).
Behavioral Coupling in (Orthoptera)

Studies of the mating behavior of male and female F1 hybrids between closely related taxa can provide information concerning the genetic control of characters that play a major role in speciation. Orthoptera have been used previously for such studies. Hybrid crickets show behaviors which are broadly intermediate to the parentals but hybrid grasshoppers may retain parental behavior patterns. This study examines the behavior of hybrid Ephippiger ephippiger bushcrickets, the third major orthopteran group. The differences in male song and female preference are probably both mainly additive and male song differences not sex linked. Thus, given a choice, hybrid females would prefer to mate with hybrid males, an example of "'behavioral coupling. "" The evolutionary inferences which can be drawn from studies of F1 hybrids between closely related taxa are discussed.
New Genes for Epilepsy–Autism Comorbidity

Abstract Similar genetic abnormalities have been demonstrated for epilepsy and autism spectrum disorder (ASD). Genetic syndromes, such as fragile X syndrome and tuberous sclerosis, also have high rates of comorbidity with epilepsy and ASD. Furthermore, next-generation sequencing has identified common genetic abnormalities in epilepsy and ASD, and recurrent de novo gene alterations can be recognized as mutations if there is sufficient evidence for pathogenicity. Such mutations occur in SCN2A and GRIN2B. SCN2A encodes the α2 subunit of the Nav1.2 neuronal voltage-gated Na+ channel, which is mutated in several genetically mediated epilepsies. GRIN2B encodes a subtype of the N-methyl-D-aspartate glutamate receptor and is mutated in epileptic encephalopathies, which often present with features of ASD. In addition, KCNT1 encodes the Na+-dependent K+ channel known as Slack. Mutations in KCNT1 produce several epilepsy phenotypes, including autosomal dominant frontal lobe epilepsy, which often includes features of ASD. Slack also has an intracellular fragile X mental retardation protein-binding domain, which is significant in its relationship with intellectual disabilities. Furthermore, DEPDC5 mutations produce autosomal dominant familial focal epilepsy with variable foci and may contribute to ASD. DEPDC5 encodes a regulator of the mechanistic target of rapamycin (mTOR) pathway, which is also affected by tuberous sclerosis mutations, suggesting that “mTORopathy” may underlie both epilepsy and ASD. Mutations in PCDH19 also produce an epilepsy syndrome that exclusively affects heterozygous females and is associated with ASD. The genetic similarities between ASD and epilepsy provide insight into the pathological molecular mechanisms of both conditions and therefore may be useful for understanding clinical outcomes.
Cellular expression of two key water borne pathogens towards antimicrobial quaternized chitosan

The antibacterial activity of chitosan and its quarternized derivative i.e., TMC against Salmonella typhimurium and Shigella flexneri was evaluated and an attempt to understand the mechanism of action was attempted in this study. The two key gram-negative water borne pathogens was evaluated for viability at different incubation times. Further, the integrity of the cell membranes of both bacterial cells were investigated by determining the release from cells of materials that absorb at 260 nm, electric conductivity measurements and leakage release of cytoplasmic β-galactosidase activity. Results revealed that after treatment with TMC, the electric conductivity of bacterial suspensions increased, followed by increasing of the units of average release for alkaline phosphatase (ALP) and glucose-6-phosphate dehydrogenase (G6PDH). SDS PAGE and occurrence of protein in cell free supernatant indicated that the soluble proteins decreased or disappeared in the treated Salmonella cells, demonstrating that TMC performed its antibacterial function via increasing the permeability of cell membranes. The results obtained with two water borne pathogens demonstrate the complexity of the mode of action of Quaternized biopolymer. An explanation of the antibacterial mechanism is proposed involving the cell wall disruption due to interaction between positive amino groups present in the TMC with the negatively charged bacterial cell surface. Several lines of evidence suggest that its site of action is the outer membrane, but a probability of more than one target site would explain quaternized chitosan’s antimicrobial action.
Concordant and divergent strategies to improve postprandial glucose and lipid metabolism

Following ingestion of a mixed-macronutrient meal, blood glucose and triacylglycerol (TAG) concentrations are elevated for approximately 2 and 8 hours, respectively, depending on factors such as the amount and type of food ingested. For many individuals who consume at least three meals per day, the majority of waking hours are thus spent in a postprandial state. Exaggerated postprandial glycaemia and triglyceridemia are associated with cardiovascular and metabolic diseases, and so strategies to alleviate such perturbations are desirable (e.g. exercise and/or diet). An acute bout of endurance, resistance or high-intensity and interval exercise typically mitigates postprandial glucose and TAG responses to subsequent meals. Regular exercise training also facilitates adaptations that beneficially alter postprandial metabolism. The health benefits of various dietary protocols have been debated in research and in the media over recent decades. To complicate matters, these strategies may be beneficial for glucose metabolism but simultaneously impair TAG metabolism or vice versa. Finally, despite the finding that diet and exercise can independently alter postprandial metabolism, there are fewer investigations of their ability to act synergistically to achieve a prophylactic response on postprandial blood glucose and TAG concentrations. This may provide the basis for future public health guidelines and is an important consideration in the context of the current worldwide prevalence of cardiovascular and metabolic diseases.
Características antropométricas e de aptidão física de meninos atletas de futebol DOI: 10.5007/1980-0037.2010v12n4p239

O presente estudo teve por objetivo determinar as caracteristicas antropometricas e de aptidao fisica de meninos no inicio da fase de treinamento especializado em futebol. Foram avaliados 282 meninos atletas de 10 a 13 anos de idade. Os meninos atletas estavam na fase inicial do treinamento especializado, 3 vezes/semana, 3 horas/dia. Todos foram submetidos a testes e medidas antropometricas e de aptidao fisica. Os meninos atletas foram agrupados em faixas etarias e os dados relativos as prevalencias nas diferentes categorias antropometricas e de composicao corporal foram analisados por meio do teste do qui-quadrado de Pearson. Os dados parametricos foram comparados por meio da ANOVA one-way e, quando necessario, por meio do metodo de Kruskal-Wallis. Todos os resultados foram expressos como media±desvios-padrao e os resultados foram considerados estatisticamente diferentes quando p<0,05. O crescimento, o desenvolvimento, a adiposidade corporal e a aptidao fisica foram adequadas e proporcionais a idade dos meninos atletas de futebol (p<0,05). Concluiu-se que as caracteristicas antropometricas e de aptidao fisica de meninos das categorias de base da elite do futebol melhoram e sao proporcionais a idade. Criancas e adolescentes atletas de futebol exibem favoraveis caracteristicas antropometricas e de aptidao fisica.
Establishment of Methods for Isolation of Pnmt+ Cardiac Progenitor Cells

Cardiovascular disease is the leading cause of death in the United States. Millions of patients suffer each year from endothelial dysfunction and/or debilitating myocardial damage resulting in decreased quality of life and increased risk of death or disablement. Current pharmacological approaches are only partly effective at treating cardiovascular disease, and hence, better strategies are needed to provide significant improvements in treatment options. Cardiac stem/progenitor cells have the potential to regenerate myocardial tissue and repair damaged heart muscle. There are many different types of cardiac progenitor cells, and each may have certain unique properties and characteristics that would likely be useful for particular clinical applications. A current challenge in the field is to identify, isolate, and test specific cardiac stem/progenitor cell populations for their ability to repair/regenerate myocardial tissue. Our laboratory has discovered a new type of cardiac progenitor cell that expresses the enzyme, Phenylethanolamine-n-methyltransferase (Pnmt). My initial studies focused on identification of Pnmt+ cells based on knock-in of a nuclear-localized Enhanced Green Fluorescent Protein (nEGFP) reporter gene into exon 1 of the Pnmt gene in a stable recombinant Pnmt-nEGFP mouse embryonic stem cell (mESC) line. These cells were differentiated into cardiomyocytes, and I identified nEGFP+ cells using fluorescence, immunofluorescence, and phase-contrast microscopy techniques. Our results showed that only about 0.025% ( 1 per 4000) of the cardiac-differentiating stem cells expressed the nEGFP+ marker. Because of the relative rarity of these cells, optimization of isolation methods proved initially challenging. To overcome this technical barrier, I used a surrogate cell culture system to establish the methods of isolation based on expression of either a fluorescent cell marker (EGFP), or a unique cell surface receptor represented by an inactivated (truncated) version of the human low-affinity nerve growth factor receptor (LNGFR). Plasmid DNA containing these reporter genes was transiently transfected into a permissive cell line (RS1), and reporter gene expression was used to identify
Impact of climate change and race evolution on the epidemiology and ecology of stripe rust in central and eastern USA and Canada

Abstract Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most devastating diseases of wheat globally. Since 2000, the geographic footprint of Pst has increased in North America due to a combination of changing weather patterns and introductions of new and more virulent strains. Pst is endemic throughout North America, but the emergence of new virulent strains poses an increased risk to wheat cultivation. As climate changes, so will the overwintering and over-summering range of this pathogen, resulting in earlier and more severe disease epidemics in the central and eastern USA and Canada. This perspective aims to highlight the changes taking place in Pst ecology and epidemiology over the last decade in North America, and then presents recommendations for future research to prepare for the changing dynamics of stripe rust. Specifically, to manage stripe rust in the future, molecular surveillance mechanisms need be established to identify and track new virulent races, and to obtain a deeper understanding of the adaptive ability of Pst. This will allow for a better explanation of how new races of Pst emerge, and encourage the development of improved epidemiological models and durable resistance strategies.
FRAKSI ETIL ASETAT KULIT BATANG FALOAK (Sterculia quadrifida R.Br ) MENGINDUKSI APOPTOSIS DAN SIKLUS SEL PADA SEL KANKER PAYUDARA T47D

Faloak (Sterculia quadrifida R.Br) is one source of bioactive compounds that could be developed as chemotherapeutic agent. Empirically East Nusa Tenggara people use boiled water of faloak bark as a cure for hepatitis (types A, B, and C), and gastroenteritis. This study was performed to test anticancer activity fraction of n-hexane, ethyl acetate, ethanol, and methanol of ethanolic extract from faloak stem bark for the type of breast cancer cell line T47D, and normal cell types Vero using cytotoxic 3- (4,5-dimetilazol- 2-yl) -2,5-difeniltetrazolium bromide (MTT) test method. Ethanolic extract was subjected to column chromatography using different solvents polarity level as n-hexane, ethyl acetate, ethanol, and methanol. Testing the cytotoxic effects using the MTT assay in T47D breast cancer cells and normal Vero cells with EC50 parameter. Ethyl acetate fraction in inducing apoptosis and cell cycle modulation was observed with flowcytometry method. The test results cytotoxic fraction indicating the fraction of ethyl acetate has the lowest activity with EC50 of 24.88 μg/mL and selectivity index of 15.58. Ethyl acetate fraction effects an accumulation of cells in S phase (27.43%) in breast cancer cells T47D which is able to induce apoptosis. These results demonstrate that the ethyl acetate fraction can be developed as a chemotherapeutic agent in improving the effectiveness of breast cancer treatment.
Studies on polymorphism of CXCR1 gene and its association with subclinical mastitis in Indian water buffalo

Sub clinical mastitis (SCM) in buffalo is considered as multi-factorial disease, which leads to huge economic losses by reducing the quality and quantity of the milk. Although bacteria are considered to be the main causative agent for mastitis, farm management practices, genetic make-up of the animal are found to be associated with the disease. So the present study aimed at identification of SNPs in the bovine CXCR1 through PCR-SSCP, DNA sequencing and its association if any with mastitis tolerance/succeptibility in Indian water buffalo. Animal wise the prevalence of sub clinical mastitis was found to be (27. %), and 7.9%, 9.9%, 2.9% quarters were found to be affected with latent mastitis, specific mastitis and nonspecific mastitis respectively . On the basis of individual quarter, the highest prevalence was in left hind (33.5%), followed by right hind (29.7%) and right front (22.2%) and the least prevalence in left front (14.6%). The microorganisms isolated from sub clinical mastitis constituted Staphylococci (39%), Streptococci (31%), Corynebacterium (25%) and E.coli. (5%). PCR-SSCP when used to explore polymorphism in partial exon 2 of CXCR1 gene, the 311bp fragment of this gene was found to be monomorphic in all the DNA samples screened. SSCP analysis of the exonic amplicons and DNA sequence analysis of CXCR1 gene exonic sequences (using one set of PCR primers) did not reveal any polymorphism in this region of CXCR1 gene in diseased or healthy buffaloes. Farm management associated determinants (type of feed, type of floor, education of the owner, feeding after milking, type of labor) when studied in association with sub clinical mastitis, education of the owner , type of labor and feeding after milking were found to be significantly associated with disease occurrence (p<0.05). While looking at the animal determinants in association with sub clinical mastitis in the present investigation, buffaloes with higher parity showed more risk of developing sub clinical mastitis. So the present investigation helped to understand the various risk factors for the occurrence of sub clinical mastitis in buffaloes in Punjab.
Influence of Transgenic Corn Expressing Insecticidal Protein of Bacillus thuringiensis Berliner on Natural Populations of Corn Earworm (Lepidoptera: Noctuidae) and Southwestern Corn Borer (Lepidoptera: Crambidae)

A 2-yr study was conducted to measure the influence of transgenic corn, Zea mays L. , expressing the CrylAb endotoxin of Bacillus thuringiensis (Berliner) (Bt) by means of Event MON810 on natural populations of Helicoverpa zea (Boddie) and Diatraea grandiosella (Dyar). The studies were conducted at Leland and Morgan City, MS, in 1999 and at Morgan City in 2000. Although total numbers of H. zea larvae were not significantly different on transgenic corn hybrids compared with their near-isogenic parent lines, fewer large larvae were found on the transgenic hybrids. Differences in H. zea larval growth were noticeable when larvae fed on Bt corn vs non-Bt corn. The delay in larval growth for insects within a single generation, which could possibly result in asynchronous mating between insecticide resistant and susceptible insects, was observed for larvae feeding on plants expressing the Bt toxin. Diatraea grandiosella caused limited damage to the transgenic corn hybrids compared with their near-isogenic parent l.
Management of black scurf disease of potato in Rajasthan

Black scurf (Rhizoctonia solani) diseases of potato are becoming prominent in many potatoes growing state resulting in economic losses, since a decade. An experiment was conducted to find out more effective management approache through chemicals, taking cv. Kufri Bahar with five treatments involving chemicals i. e. Pencycuron (Moncern) and Boric acid, application before storage alone. Minimum disease incidence (17.25) and tuber yield (26.11 t/ha) was noticed in Tuber spray treatment with Pencycuron 22.9% (Moncern) @ 0.50 before planting alone followed by Tuber dip treatment with 3% boric acid or 10 min. before planting. These treatments can provide an effective management of potato tuber diseases
Effects of some physicochemical properities of water on histopathological and hematological picture of fish inhabiting Emet Stream

This study deals with the physicochemical water parameters of Emet Stream and their histopathologic and hematologic effects upon Squalius cephalus, Capoeta tinca, Barbus plebejus and Alburnoides bipunctatus living there. The changes of the fish tissues were determined by the use of routine histological methods. The hematocrit levels of the bloods taken from the fish were also estimated. NO2, PO4 3-, Ni, as and B values were observed to be above the stipulated levels. There were bronchitis, telangiectasia, epithelial lifting and passive hyperemia determined in gills; focal hepatitis, vena centralis, hydropic and vacuolar degeneration were seen in livers; there were hemorrhages in kidney tissue and enteritis was encountered. Bronchitis and telangiectasis were intensively observed in S. cephalus and C. tinca specimens. Focal hepatitis was much more intensive in C. tinca. The hematocrit levels were foundrnto be 520.45% in S. cephalus, 17.55% in A. bipunctatus, 16.625% in B. plebejus and 22% in C. tinca. The elevated values of some water parameters of Emet Stream were actually affecting on both tissues and blood parameters of the fish.
Male biased sex ratio reduces the fecundity of one of three female morphs in a polymorphic damselfly

Lay SummaryHow genetic variation is preserved is a central issue in evolutionary biology. In many species of damselflies, females come in distinct heritable color varieties. We found that in an Iberian damselfly when subject to stressful male harassment, fecundity decreased in one kind of female but not in others. Our results, together with previous observations, suggest a scenario where differential fecundity, male preference, and fluctuations in male abundance play an important role in maintaining this variation.
Targeted Nucleotide Substitution in Mammalian Cell by Target-AID

[Abstract] Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems have been applied to various type of cells as powerful genome editing tools. By using activation-induced cytidine deaminase (AID) in place of the nuclease activity of the CRISPR/Cas9 system, we have developed a genome editing tool for targeted nucleotide substitution (C to T or G to A) without donor DNA template (Figure 1; Nishida et al., 2016). Here we describe the detailed method for Target-AID to perform programmable point mutagenesis in the genome of mammalian cells. A specific method for targeting the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene in Chinese Hamster Ovary (CHO) cell was described here as an example, while this method principally should be applicable to any gene of interest in a wide range of cell types.