Fasciola hepatica surface tegument: glycoproteins at the interface of parasite and host

Fasciola hepatica, commonly known as liver fluke, is a trematode which causes Fasciolosis in ruminants and humans. The outer tegumental coat of F. hepatica (FhTeg) is a complex metabolically active biological matrix that is continually exposed to the host immune system and therefore makes a good vaccine target. F. hepatica tegumental coat is highly glycosylated and helminth-derived immunogenic oligosaccharide motifs and glycoproteins are currently being investigated as novel vaccine candidates. This report presents the first systematic characterisation of FhTeg glycosylation using lectin microarrays to characterise carbohydrates motifs present, and lectin histochemistry to localize these on the F. hepatica tegument. We discovered that FhTeg glycoproteins are predominantly oligomannose oligosaccharides that are expressed on the spines, suckers and tegumental coat of F. hepatica and lectin blot analysis confirmed the abundance of N- glycosylated proteins. While some oligosaccharides are widely distributed on the fluke surface other subsets are restricted to distinct anatomical regions. We selectively enriched for FhTeg mannosylated glycoprotein subsets using lectin affinity chromatography and identified 369 proteins by mass spectrometric analysis. Among these proteins are a number of potential vaccine candidates with known immune modulatory properties including proteases, protease inhibitors, paramyosin, Venom Allergen-like II, Enolase and two proteins, nardilysin and TRIL, that have not been previously associated with F. hepatica Furthermore, we provide a comprehensive insight regarding the putative glycosylation of FhTeg components which could highlight the importance of further studies examining glycoconjugates in host-parasite interactions in the context of F. hepatica infection and the development of an effective vaccine.

Exploring the cardiovascular disease continuum: blood pressure and target organ damage

Introduction The objectives of this thesis are to: (1) examine how ambulatory blood pressure monitoring (ABPM) refines office blood pressure (BP) measurement; (2) determine if absolute ambulatory BP or dipping status is better associated with target organ damage (TOD); (3) explore the association of isolated nocturnal hypertension (INH) with TOD; and (4) investigate the association of night-time BP with ultrasound markers of cardiovascular damage. Methods Data from the Mitchelstown Cohort Study was analysed to deliver objectives 1 and 2. Objective 3 was addressed by a systematic review and analysis of data from the Mitchelstown Study. A sample of participants from the Mitchelstown Study underwent an echocardiogram for speckle tracking analysis and carotid ultrasound to achieve objective 4. Results ABPM reclassifies hypertension status in approximately a quarter of individuals, with white coat and masked hypertension prevalence rates of 11% and 13% respectively. Night-time systolic BP is better associated with TOD than daytime systolic BP and dipping level. In multi-variable models the odds ratio (OR) for LVH was 1.4 (95% CI 1.1 -1.8) and for albumin:creatinine ratio ≥ 1.1 mg/mmol was 1.5 (95% CI 1.2 – 1.8) for each 10 mmHg rise in night-time systolic BP. The evidence for the association of INH with TOD is inconclusive. Night-time systolic BP is significantly associated with global longitudinal strain (GLS) (beta coefficient 0.85 for every 10 mmHg rise, 95% CI 0.3 – 1.4) and carotid plaques (OR 1.9 for every 10 mmHg rise, 95% CI 1.1 – 3.2) in univariable analysis. The findings persist for GLS in sex and age adjusted models but not in multivariable models. Discussion Hypertension cannot be effectively managed without using ABPM. Night-time systolic BP is better associated with TOD than daytime systolic BP and dipping level, and therefore, may be a better therapeutic target in future studies.

ENERGETIC BIOCIDAL MATERIALS AND SPORE NEUTRALIZATION

The objective of this dissertation is to explore a more accurate and versatile approach to investigating the neutralization of spores suffered from ultrafast heating and biocide based stresses, and further to explore and understand novel methods to supply ultrafast heating and biocides through nanostructured energetic materials A surface heating method was developed to apply accurate (± 25 ˚C), high heating rate thermal energy (200 - 800 ˚C, ~103 - ~105 ˚C/s). Uniform attachment of bacterial spores was achieved electrophoretically onto fine wires in liquids, which could be quantitatively detached into suspension for spore enumeration. The spore inactivation increased with temperature and heating rate, and fit a sigmoid response. The neutralization mechanisms of peak temperature and heating rate were correlated to the DNA damage at ~104 ˚C/s, and to the coat rupture by ultrafast vapor pressurization inside spores at ~105 ˚C/s. Humidity was found to have a synergistic effect of rapid heating and chlorine gas to neutralization efficiency. The primary neutralization mechanism of Cl2 and rapid heat is proposed to be chlorine reacting with the spore surface. The stress-kill correlation above provides guidance to explore new biocidal thermites, and to probe mechanisms. Results show that nano-Al/K2S2O8 released more gas at a lower temperature and generated a higher maximum pressure than the other nano-Al/oxysalts. Given that this thermite formulation generates the similar amount of SO2 as O2, it can be considered as a potential candidate for use in energetic biocidal applications. The reaction mechanisms of persulfate and other oxysalts containing thermites can be divided into two groups, with the reactive thermites (e.g. Al/K2S2O8) that generate ~10× higher of pressure and ~10× shorter of burn time ignited via a solid-gas Al/O2 reaction, while the less reactive thermites (e.g. Al/K2SO4) following a condensed phase Al/O reaction mechanism. These different ignition mechanisms were further re-evaluated by investigating the roles of free and bound oxygen. A constant critical reaction rate for ignition was found which is independent to ignition temperature, heating rate and free vs. bound oxygen.

Improving interfaces for nerve repair

Autologous nerve grafts are the current gold standard for the repair of peripheral nerve injuries. However, there is a need to develop an alternative to this technique, as donor-site morbidities such as neuroma formation and permanent loss of function are a few of the limitations concerned with this technique. Artificial nerve conduits have therefore emerged as an alternative for the repair of short peripheral nerve defects of less than 30 mm, however they do not surpass autologous nerve grafts clinically. To develop a nerve conduit that supports regeneration over long nerve gaps and in large diameter nerves, researchers have focused on functionalizing of the conduits by studying the components that enhance nerve regeneration such as micro/nano-topography, growth factor delivery systems, supportive cells and extracellular matrix (ECM) proteins as well as understanding the complex biological reactions that take place during peripheral nerve regeneration. This thesis presents strategies to improve peripheral nerve interfaces to better the regenerative potential by using dorsal root ganglions (DRGs) isolated from neonatal rats as an in vitro model of nerve regeneration. The work started off by investigating the usefulness of a frog foam protein Ranaspumin-2 (Rsn2) to coat biomaterials for compatibility, this lead to the discovery of temporary cell adhesion on polydimethylsiloxane (PDMS), which was investigated as a suitable tool to derive cell-sheets for nerve repair. The influence of Rsn2 anchored to specific adhesion peptide sequences, such as isoleucine-lysine-valine-alanine-valine (IKVAV), a sequence derived from laminin proven to promote cell adhesion and neurite outgrowth, was tested as a useful means to influence nerve regeneration. This approach improves the axonal outgrowth and maintains outgrowth long term. Based on the hypothesis that combinational modulation of substrate topography, stiffness and neurotrophic support, affects axonal outgrowth in whole DRGs, dissociated DRGs were used to assess if these factors similarly act at the single cell level. Rho associated protein kinase (ROCK) and myosin II inhibitors, which affect cytoskeletal contractility, were used to influence growth cone traction forces and have shown that these factors work in combination by interfering with growth cone dynamic creating a different response in axonal outgrowth at the single cell level.

Determinación de cortisol plasmático en pacientes tuberculosos y en personas sanas luego de la estimulación con ACTH sintético.1992-1993

Estudio cuasiexperimental en 50 pacientes tuberculosos ingresados en el Hospital Neumológico y 50 voluntarios aparentemente sanos de la ciudad de Cuenca, para determinar el cortisol basal y sus valores a las 8. 24 y 36 horas luego de la estimulación con ACTH (Synacthen Depot) a la dosis de 0.5 miligramos vía muscular. Todos los individuos de estudio tuvieron entre 20 y 50 años y no haïbían recibido medicación corticoide por lo menos 15 días antes del estudio. La determinación del cortisol se hizo por radioinmunoanálisis con Coat-A Count Cortisol, siguiendo los procedimientos indicados por el laboratorio. Las cifras de cortisol basal en los individuos sanos son de 13.94 +- 5.78 ug/dl. y en los tuberculosos de 17.83+- 6.75 ug/dl, siendo la diferencia estadísticamente significativa (p menor que 0.001) Considerando los sexos, los valores de cortisol entre hombres y mujeres no son estadísticamente diferentes. A las 8 horas luego de la estimulación con ACTH el 90de los individuos aparentemente sanos y el 74de los tubeculosos presentan una respuesta adecuada a la estimulación, ya que duplican o triplican los valores basales. En la determinación de cortisol a las 24 horas un 46de individuos en ambos grupos no llegan a duplicar los valores y a las 36 horas únicamente duplican el 26de los individuos salos y el 20de los tuberculosos por lo que se hace innecesaria su determinación. Considerando que en los tuberculosos las cifras están elevadas y que esta enfermedad suele acompañarse de deficiencias nutritivas, se recomienda realizar estudios en pacientes con diferentes grados y tipos de desnutrición

Cambios fisiológicos y bioquímicos en cirugía laparoscópica vs convencional de vías biliares. Hospital Vicente Corral Moscoso Cuenca 2000

Con un diseño cuasiexperimental se realizó un estudio clínico en el hopital Vicente Corral de Cuenca que incluyó 80 pacientes de ambos sexos asignados a dos grupos: 1.- grupo COLELAP formado por cuarenta pacientes a los que se realizó colecistectomía laparoscópica y 2.- grupo CONVENC formado por 40 pacientes a los que se realizó colecistectomía abierta. El procedimiento fue similar para ambos grupos. En los dos grupos se midieron a.- los niveles de cortisol prequirúrgico y posquirúrgico mediante radioinmunoanálisis [Coat-A cortisol r], b.- El dolor antes y después de la cirugía mediante una escala visual y c.- La estadía hospitalaria y las complicaciones. No hubieron diferencias estadísticamente significativas en cuanto a edad, género, procedencia, nivel de instrucción, dolor preoperatro y valoraciones sanguíneas de laboratorio. Los valores de cortisol del grupo COLELAP fueron inferiores a los del grupo CONVENC tanto en el preoperatorio 9.40 más menos 4.81 vs. 20.11 más menos 8.87 [p = 0.001] como en el postoperatorio 12.19 más menos 4.12 vs. 22.1 más menos 7.31 [p=0.0001], sin embargo en el grupo COLELAP el aumento promedio [2.8 mcg/dL vs 1.99 mcg/dL fue mayor. El dolor preoperatorio evaluado mediante una escala análoga visual fue igual en ambos grupos [p=0.22] pero en el postoperatorio fue menor en el grupo COLELAP [p=0.012]. El tiempo quirúrgico promedio fue mayor para el grupo CONVENC [82.25 más menos 31.66 min vs 63.5 más menos 31.23 min] [p=0.0005]. El 90 por ciento de los pacientes del grupo fue de 3.1 más menos días y para el grupo CONVENC de 4.8 más menos 0.87 días. La diferencia fue altamente significativa [p=0.00001]. En conclusión laparoscópica muestra significativas ventajas frente a la colecistectomía abierta; a.- Minimiza el trauma quirúrgico, b.- produce menor dolor postoperatorio y c.- disminuye la estadía hospitalaria

Anticorpos anti-trypanosoma cruzi como preditivos de infecção por leishmania infantum

Leishmania infantum and Trypanosoma cruzi are trypanosomatids of medical importance and are, respectively, the etiologic agents of visceral leishmaniasis (VL) and Chagas disease (CD) in Brazil. People infected with L. infantum or T. cruzi may develop asymptomatically, enabling the transmission of pathogens through blood transfusion and / or organs. The assessment of the infection by T. cruzi is included among the tests performed for screening blood donors in Brazil, however, there is no availability of tests for Leishmania. Serological tests for T. cruzi are very sensitive, but not specific, and may have cross-reactions with other microorganisms. Thus, the aim of this study was to determine the prevalence of Leishmania infection in blood donors and assess whether the serological test for T. cruzi detect L. infantum. Among the 300 blood samples from donors, discarded in 2011, 61 were T. cruzi positive, 203 were from donors with other infections and 36 were from handbags with low blood volume, but without infection. We also assessed 144 samples from donors without infections and able to donate blood, totaling 444 subjects. DNA was extracted from blood samples of all to perform quantitative PCR (qPCR) to detect Leishmania DNA. The buffy coat obtained from all samples was grown in Schneider medium supplemented and NNN. All samples were evaluated for the presence of anti-Leishmania antibody. The serological results indicate a percentage of 22% of Leishmania infection in blood samples obtained from discarded bags. A total of 60% of samples positive in ELISA for T. cruzi were negative by IFI, used as confirmatory test, ie 60% false positive for Chagas. Among these samples false positive for Chagas, 72% were positive by ELISA for Leishmania characterizing the occurrence of cross reaction between serologic assays. Of the 300 cultures performed, 18 grew parasites that were typed by qPCR and specific isoenzymes, found the species Leishmania infantum crops. Among the 18 cultures, 4 were purged from scholarships for low volume and all negative serology blood bank, thus demonstrating that there is a real risk of Leishmania transmission via transfusion. It is concluded that in an area endemic for leishmaniasis in Brazil, serological diagnosis performed to detect infection by T. cruzi among blood donors can identify infection by L. infantum and although cause false positive for Chagas, this cross-reactivity reduces the risk of Leishmania infection via blood transfusion, since tests are not applied specific detection of the parasite. In this way, there remains the need to discuss the implementation of a specific serological screening test for Leishmania in endemic countries such as Brazil

Copy from actual survey of the 3 Sisters : [Potomac River, Washington D.C.] /

Cadastral map.

Variações anátomo-fisiológicas face à aclimatação sazonal - estudo em vacas leiteiras com diferente potencial leiteiro

O presente estudo teve como objetivo a caracterização de variações anátomo-fisiológicas que decorrem do processo de aclimatação sazonal em bovinos leiteiros com alto (Altas) e baixo (Baixas) potencial leiteiro, com vista a selecionar animais que conciliem bons desempenhos produtivos e índices de adaptabilidade que possibilitem a mudança de homeostase face às condições climáticas presentes no clima mediterrânico. O estudo foi realizado numa herdade comercial, situada no Alentejo, utilizando 13 vacas multíparas (6 Baixas e 7 Altas), durante três períodos: P1 (animais aclimatados ao verão; presença de stresse térmico); P2 (animais aclimatados ao verão; termoneutralidade); P3 (animais aclimatados ao inverno; termoneutralidade). Em stresse térmico (P1), verificaram-se maiores esforços termolíticos e maiores armazenamentos de calor no grupo das Altas. Observou-se também que a produção de leite das Altas foi afetada pelo stresse térmico, evidênciando uma redução 24-48h após os valores de temperatura retal mais elevados. Nesta situação, as Baixas apresentaram uma variação na produção oposta à das Altas. Em P1, os valores de proteína e de gordura no leite foram significativamente mais baixos que em P3, em ambos os grupos. A ureia no leite foi significativamente mais elevada nas Altas durante o P1, revelando potencial como biomarcador de stresse térmico. Do P1 para o P3 obser-vou-se uma redução gradual do hematócrito, da hemoglobina e da triiodotironina (T3). As Altas apresentaram uma maior redução de triiodotironina (T3) que as Baixas, como consequência de uma maior intensidade de aclimatação. Nos pelos não se registaram diferenças entre os períodos, o que contrasta com alguma bibliografia. Porém, a ausência da insolação direta poderá ter sido um fator determinante; ABSTRACT: The main objective of the present study was the characterization of anatomical and physiological variations that occur in the seasonal acclimatization process of dairy cows with high (Altas) and low (Baixas) milk yield potential. In this way it should be possible to do a selection of animals with good productive traits and also with adaptability indexes that allow a change in homeostasis to cope with the climatic conditions of the mediterranean climate. Meteorological, clinical, productive, physiological and anatomical data were collected. The study was conducted in an Alentejo's dairy farm, using 13 multiparous cows (6 with low milk yield and 7 with high milk yield), during three periods: P1 (animals acclimated to summer, in heat stress); P2 (animals acclimated to summer, thermoneutrality); P3 (animals acclimated to winter; thermoneutrality). In thermal stress (P1), the high milk yield group (Altas) shown greater thermolytic efforts and also higher heat storage. The milk yield in this group was also affected by heat stress, showing a decrease in production when the rectal temperature increased, with a delay of 24-42 hours. In this situation the Baixas group showed an opposite milk production variation. In P1, the protein and fat milk content was lower than in P3, in both groups. Milk urea levels were significantly higher during P1 in the Altas group, revealing potential as an heat stress biomarker. Hematocrit, hemoglobin and triiodothyronine (T3) values gradualy decreased from P1 to P3. T3 values were lower in Altas than in Baixas group, as a consequence of a more intense acclimatization. The hair analysis didn’t show the standard seasonal acclimatization process, indicating the absence of direct solar radiation as a determinant factor.

Síntese verde de nanopartículas de prata a partir de extrato aquoso do tubérculo de Curcuma longa associadas à quitosana e avaliação da atividade antitumoral in vitro em câncer de pele não melanoma (linhagem A431)

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação Nanociência e Nanobiotecnologia, 2016.

Nutrient content in aboveground biomass of Brazilian peanut cultivars in conservation tillage on sugarcane straw and pasture area.

Fields studies were conducted in 2004/2005 in order to evaluate the effects of tillage on nutrient content in aboveground biomass of two peanut cultivars, cultivated in rotation after mechanical harvested sugarcane and pastures. These trials were carried out in two types of soils; Oxisol and Ultisol, respectively in Ribeir?ao Preto and Mirassol, S?ao Paulo State, Brazil. The experimental design was split-plot with four replications. Tillage treatments (conventional, minimum and no-tillage) were main plots while sub-plots were peanut genotypes IAC-Tatu ST (Valencia market-type, erect growth habit, red seed coat, maturity range around 100 days after planting) and IAC-Caiap´o (Runner market-type, prostate growth habit, pink testa, maturity range more than 135 days). From 15 to 90 days after emergence, samples of leaves and stems were harvested, dried, weighted and ground to determine macro and micronutrient concentration. At 75 days after sowing, the cultivar IAC-Caiap´o showed higher contents of N, P, K, Cu, and Zn while IAC-Tatu presented higher concentrations of Ca, Mg, and S. Zn content was higher in conservation tillage than in conventional, mainly in Oxisoil for both of cultivars.

Anatomical, histochemical and immunohistochemical characterization of the outflow tract of ray hearts (Rajiformes; Chondrichthyes)

Recent work has shown that the cardiac outflow tract of sharks and chimaeras does not consist of a single myocardial component, the conus arteriosus, as classically accepted, but two, namely, the myocardial conus arteriosus and the non-myocardial bulbus arteriosus. However, the anatomical composition of the outflow tract of the batoid hearts remains unknown. The present study was designed to fill this gap. The material examined consisted of hearts of two species of rays, namely, the Mediterranean starry ray (Raja asterias) and sandy ray (Leucoraja circularis). They were studied using scanning electron microscopy, and histochemical and inmunohistochemical techniques. In both species, the outflow tract consists of two components, proximal and distal with regard to the ventricle. The proximal component is the conus arteriosus; it is characterized by the presence of compact myocardium in its wall and several transverse rows of pocket-shaped valves at its luminal side. Each valve consists of a leaflet and its supporting sinus. Histologically, the leaflet has two fibrosas, inner and outer, and a middle coat, the spongiosa. The distal component lacks myocardium. Its wall consists of smooth muscle cells, elastic fibers and collagen. Thus, it shows an arterial-like structure. However, it differs from the aorta because it is covered by the epicardium and crossed by coronary arteries. These findings indicate that the distal component is morphologically equivalent to the bulbus arteriosus of sharks and chimaeras. In contrast to foregoing descriptions, the valves of the first transverse row are distally anchored to the bulbus arteriosus and not to the ventral aorta. Our findings give added support to the notion that presence of a bulbus arteriosus at the arterial pole of the heart is common to all chondrichtyans, and not an apomorphy of actinopterygians as classically thought.

Detection of Rupestris stem pitting associated virus in seedlings of virus - infected maternal grapevine plants.

Rupestris stem pitting associated virus (RSPaV) is a species in the genus Foveavirus (Martelli and Jelkman, 1998) and the family Flexiviridae. The virion has a positive sense, single stranded, polyadenylated RNA genome of 8.7kb in size and a coat protein of 28kD (Martelli and Jelkman, 1998). The virus has been reported to be present in pollen (Rowhani et aI., 2000) and seeds (Stewart and Nassuth, 2001), however, it has not been proved to be seed-transmitted. In our investigation reported here we have proven that RSPaV transmits by seed from RSPaV-infected mother plants to their siblings.

The bacterial chromosome segregation protein Spo0J spreads along DNA from parS nucleation sites

Regulation of chromosome inheritance is essential to ensure proper transmission of genetic information. To accomplish accurate genome segregation, cells organize their chromosomes and actively separate them prior to cytokinesis. In Bacillus subtilis the Spo0J protein is required for accurate chromosome segregation and it regulates the developmental switch from vegetative growth to sporulation. Spo0J is a DNA-binding protein that recognizes at least eight identified parS sites located near the origin of replication. As judged by fluorescence microscopy, Spo0J forms discrete foci associated with the oriC region of the chromosome throughout the cell cycle. In an attempt to determine the mechanisms utilized by Spo0J to facilitate productive chromosome segregation, we have investigated the DNA binding activity of Spo0J. In vivo we find Spo0J associates with several kilobases of DNA flanking its specific binding sites (parS) through a parS-dependent nucleation event that promotes lateral spreading of Spo0J along the chromosome. Using purified components we find that Spo0J has the ability to coat non-specific DNA substrates. These 'Spo0J domains' provide large structures near oriC that could potentially demark, organize or localize the origin region of the chromosome.