La Albumina humana su utilización en diferentes patologías y su actual

 La albumina humana se viene utilizando desde 1945 en diferentes patologías y en los últimos años ha aumentado el número de patologías donde se usa este medicamento, los métodos de purificación a través de todos estos años se han venido mejorando y simplificando, lo que eleva su calidad y permite la reducción de los precios del medicamento. En la Universidad de Pamplona, se ha realizado el proceso completo, desde la simplificación del método de producción, sin pérdida de calidad, como la producción piloto de la albumina para análisis internacional de calidad. Los resultados obtenidos por métodos de alta fidelidad, demuestran una pureza superior al 99.98%, lo que eleva este producto de la Universidad de Pamplona a nivel internacional como un producto con capacidad competitiva en calidad y precio

IFNγ Response to Mycobacterium tuberculosis, Risk of Infection and Disease in Household Contacts of Tuberculosis Patients in Colombia

OBJECTIVES: Household contacts (HHCs) of pulmonary tuberculosis patients are at high risk of Mycobacterium tuberculosis infection and early disease development. Identification of individuals at risk of tuberculosis disease is a desirable goal for tuberculosis control. Interferon-gamma release assays (IGRAs) using specific M. tuberculosis antigens provide an alternative to tuberculin skin testing (TST) for infection detection. Additionally, the levels of IFNgamma produced in response to these antigens may have prognostic value. We estimated the prevalence of M. tuberculosis infection by IGRA and TST in HHCs and their source population (SP), and assessed whether IFNgamma levels in HHCs correlate with tuberculosis development. METHODS: A cohort of 2060 HHCs was followed for 2-3 years after exposure to a tuberculosis case. Besides TST, IFNgamma responses to mycobacterial antigens: CFP, CFP-10, HspX and Ag85A were assessed in 7-days whole blood cultures and compared to 766 individuals from the SP in Medellín, Colombia. Isoniazid prophylaxis was not offered to child contacts because Colombian tuberculosis regulations consider it only in children under 5 years, TST positive without BCG vaccination. RESULTS: Using TST 65.9% of HHCs and 42.7% subjects from the SP were positive (OR 2.60, p<0.0001). IFNgamma response to CFP-10, a biomarker of M. tuberculosis infection, tested positive in 66.3% HHCs and 24.3% from the SP (OR = 6.07, p<0.0001). Tuberculosis incidence rate was 7.0/1000 person years. Children <5 years accounted for 21.6% of incident cases. No significant difference was found between positive and negative IFNgamma responders to CFP-10 (HR 1.82 95% CI 0.79-4.20 p = 0.16). However, a significant trend for tuberculosis development amongst high HHC IFNgamma producers was observed (trend Log rank p = 0.007). DISCUSSION: CFP-10-induced IFNgamma production is useful to establish tuberculosis infection prevalence amongst HHC and identify those at highest risk of disease. The high tuberculosis incidence amongst children supports administration of chemoprophylaxis to child contacts regardless of BCG vaccination

Comparación de la activación y la cinética de la plasmina bufalina con la humana

The Plasminogen is the zymogene of the Plasmin, enzyme which physiologically is activated by two different enzymes, the tissue plasminogen activator and the urokinase, the plasmin is the enzyme that dissolves blood clots. In this study the human plasmin was compared to the bufaline plasmin, in the activation from the zymogene to the enzyme form as well as in the affinity to the chromogenic substrate. The two plasminogens were purified by the same chromatographies methods: affinity and ion-exchange. Furthermore, both plasminogens were activated by human urokinase. The bufaline plasmin showed more activation and affinity (1.35 mM) that the human plasmin (2.16 mM), in addition, the bufaline plasmin demonstrated a 1.5 times more affinity to the chromogenic substrate that the human plasmin. This study demonstrated that the plasminogens of several species can be purified by this method. Besides, one more time the animal�s plasmins probably to be more efficient in the dissolution of blood clots or degradation of substrates than the human plasmin. More over this study indicated that the bufaline plasmin can be used in clinical determinations of patients with cardiovascular diseases. This also reduces the determination time of fibrinolytic parameters that physicians can give, having more time to take appropriate treatment.O plasminogênio é o zymogen da plasmina, enzima ativada a nivel fisiológico pelo ativador tissular do plasminogênio e uroquinase, plasmina é a enzima responsável de dissolver o coágulo de sanguíneo. neste estudo foi comparada a plasmina humana com a plasmina búbalina em seu modo de ativação de zymogen a enzima e na afinidade substrato cromogênico. Os plasminogênio foram purificados com o mesmo método de cromatografia de afinidade e de troca iônica, e as ativações foram feitas usando uroquinase humana nos dois casos. A Búfalo plasmina mostrou maior ativação e afinidade (1.35 mM) que a plasmina humana (2.16 mM), sendo a bufalina 1.5 vezes mais afim ao substrato Cromogênico que a humana. Este estudo mostrou que o método de purificação do plasminogênios pode ser o mesmo para muitas espécies, alem disso, que as plasminas animais são mais eficientes na dissolução do coágulo o degradação de substratos que a plasmina humana. Este estudo indicou que a plasmina búfalo pode ser utilizada nos parâmetros determinados clínicamente em pacientes com problemas cardiovasculares, diminuindo o tempo de determinação destes parâmetros fibrinolíticos, que podem dar ao médico um intervalo de maior tempo para atuar.El plasminógeno es el zimógeno de la plasmina, enzima activada a nivel fisiológico por el activador tisular del plasminógeno y la urokinasa, la plasmina es la enzima encargada de disolver el coágulo sanguíneo. En este estudio se compararon la plasmina humana con la bufalina en su forma de activación de zimógeno a enzima y en la afinidad hacia el sustrato cromogénico. Los plasminógenos fueron purificados por el mismo método de cromatografías de afinidad y cambio iónico. De igual manera las activaciones se hicieron utilizando urokinasa humana en ambos casos. La plasmina bufalina demostró mayor activación y afinidad (1.35mM) que la plasmina humana (2.16 mM), siendo la bufalina 1.5 veces mas afin al sustrato cromogénico que la humana. Este estudio demuestra que el método de purificación de los plasminógenos puede ser el mismo para muchas especies, se demuestra una vez más que las plasminas animales al parecer son más eficientes en la disolución del coágulo o degradación de sustratos, que la plasmina humana. Este estudio indica que la plasmina bufalina puede ser utilizada en los parámetros que se determinan clínicamente en pacientes con problemas cardiovasculares, reduciendo el tiempo de determinación de estos parámetros fibrinolíticos, que pueden dar al médico un margen de tiempo superior para actuar

Identificación electroforética de 2-macroglobulina en plasma de ovino de pelo (Ovis aries) y búfalo (Bubalus bubalis)

Blood plasma from six different non pregnant and pregnant species, including human blood plasma, was analyzed for detection of á2-Macroglobulin (á2-M). The tropical hair sheep (Ovis aries) and the buffalo (Bubalus bubalis) were studied for the first time in Colombia. The presence of the á2-M in plasma of all the species was demonstrated by SDS 7.5% PAGE as bands of 180 kDa as well as by non-denaturing 5% PAGE with bands of 720 kDa. The tetrameric form �¿2-M (tetramerica) and the pregnancy zone protein (PZP) (dimeric) purified at 98%, as well as its corresponding bans from human plasma were used as control. The N-terminal sequence of the band of 180 kDa in Tropical hair sheep plasma was very similar to the purified human �¿2-M. The results indicated the presence of �¿2-M in blood plasma of all the species tested, while the PZP was present only in the pregnant human plasma. Both human and bovine �¿2-M became activated with the fast form by reacting with Methylamine. This Fac. demonstrates the differences in the reactivity of the animal�fs �¿2-M with primary amine as compared with the human �¿2-M. It could be necessary to unify purification methods into one method for all species, so that the sensitive domain of the �¿-macroglobulins (thiolester and bait region) receives the same treatment and grade of denaturation for all �¿2-M preparation.A traves del presente estudio se analizaron plasmas sanguineos de seis especies, incluyendo el humano tanto en estado gestante como no gestante, identificandose por primera vez en plasma, la glicoproteina �¿2-Macroglobulina (�¿2-M) de ovino de pelo (Ovis aries) y de bufalo (Bubalus bubalis). La presencia de esta proteina en el plasma sanguineo de todas las especies en estudio se demostro mediante electroforesis en gel de poliacrilamida usando sodio dodecilsulfato como agente denaturante (SDS PAGE) al 7.5% identificandose como bandas de 180 kDa y en forma no denaturante PAGE 5% como bandas de 720 kDa. Estas ultimas bandas fueron claramente intercambiables de la forma tetramerica a la forma monomerica en los ensayos electroforeticos. Como controles se usaron la �¿2-M (tetramerica) y la proteina de la zona de gestacion (PZP) (dimerica) purificadas a un 98%; asi como, las bandas de estas dos proteinas en el plasma humano. El analisis de la secuencia del dominio N-terminal de la (�¿2-M) de ovino de pelo, fue muy similar al de la proteina humana purificada. Tanto la �¿2-M humana como la bovina llegaron a ser activadas a la forma rapida por medio de la reaccion con metilamina. Lo anterior demuestra diferencias en la reactividad de las �¿2-M animales con la amina primaria cuando se comparan los resultados con la forma rapida de la �¿2-M humana. Sera necesario unificar los metodos de purificacion de esta proteina en todas las especies, de tal manera que los dominios sensibles de las �¿-macroglobulinas (tioester y region senuelo) tengan el mismo tratamiento y el mismo grado de desnaturalizacion para todas las preparaciones de �¿2-M

Increase in post-thaw viability by adding seminal plasma proteins to Sanmartinero and Zebu sperm

Background: cryopreservation decreases sperm viability by approximately 50%. Objective: the objective of this study was to determine the effect of the addition of seminal plasma proteins on post-thawing sperm viability in Sanmartinero and Zebu semen. Methods: semen samples from 10 bulls of each breed were used, and seminal plasma was subjected to two-dimensional electrophoresis to establish the relationship between the relative amount of each protein spot and sperm viability. Then, seminal plasma was subjected to exclusion chromatography to separate the fraction containing these proteins. This fraction was added in doses of 0.5, 1.0, 1.5 and 2.0 mg, to 1 x 10(6). Sperm was thawed and incubated at 37 °C for 1 h to determine its effect on postthaw viability. Sperm were frozen using two media (citrate-fructose-yolk and Bioxcell®). Results: we found one protein spot (16.20 kDa, PI 5.5) in Sanmartinero seminal plasma that correlated (r = 0.64 p<0.001) with viability. This spot was found in 21-25 chromatography fractions. The percentage of post-thaw viable sperm increased 20% (p<0.05) at 1.0 and 1.5 mg of the fraction when sperm was frozen using citrate-fructose-yolk; it increased 25% (p<0.01) with 0.5 mg when it was frozen with Bioxcell® media. Addition of 0.5 mg of the fraction to semen cryopreserved with Bioxcell® resulted in a greater (p<0.05) percentage increase of viable sperm in Sanmartinero semen (23 ± 8.3%) compared with Zebu semen (6.0 ± 2.0%). Conclusions: these results show that seminal plasma proteins decrease cryopreservation damage in sperm. The effect depends on the cryoprotectant dose as well as the breed of bull.Antecedentes: a criopreservação diminui a viabilidade espermática abaixo de um 50%. Objetivo: o objetivo desta pesquisa foi determinar o efeito da adição de proteínas do plasma seminal na viabilidade espermática pós-descongelamento de sêmen de touros das raças Sanmartinero y Zebú. Métodos: coletou-se sêmen de 10 touros de cada raça, as amostras do plasma seminal foram submetidas à eletroforese bidimensional, para estabelecer a relação entre a quantidade relativa de cada ponto de proteína e a viabilidade espermática. Ao serem identificados os pontos, o plasma seminal também foi submetido ao processo de cromatografia por exclusão para separar a fração que continha as proteínas. A fração foi adicionada nas doses de 0,5, 1,0, 1,5 y 2,0 mg, amostras de 1 x 106 espermatozoides, em descongelamento e incubados à temperatura de 37 ° C durante 1 hora, para determinar o efeito na viabilidade pós-descongelamento. Os espermatozoides foram congelados utilizando dois meios (Citrato- frutose-gema e Bioxcell®). Resultados: encontrou-se um ponto de proteína (16,20 kDa, ponto Isoelétrico 5,5) no plasma de touro Sanmartinero, que correlacionou (r=0,64 p<0,001) com a viabilidade. Esse ponto de proteína foi encontrado na fração 21-25 da cromatografia. O percentagem de espermatozoides viáveis pós-descongelamento aumentou em 20% (p<0,05) nas doses de 1 y 1,5 mg da fração, quando os espermatozoides foram congelados em meio de citrato-frutose-gema; e 25% (p<0,01) com doses de 0,5 mg congelados em meio Bioxcell®. A adição de 0,5 mg da fração ao sêmen descongelado e previamente criopreservado em meio Bioxcell®, evidenciou um incremento maior (p<0,05) no percentagem de espermatozoides viáveis do sêmen de touros Sanmartinero (23 ± 8,3 %), do que em sêmen de touros zebu (6,0 ± 2,0%). Conclusões: os anteriores resultados demonstram que as proteínas do plasma seminal diminuem o dano nos espermatozoides pela criopreservação e que o efeito destas proteínas depende do meio de congelação, a dose adicionada e a raça dos touros.Antecedentes: la criopreservación disminuye la viabilidad espermática por debajo del 50%. Objetivo: el objetivo de esta investigación fue determinar el efecto de la adición de proteínas del plasma seminal sobre la viabilidad espermática post-descongelación de semen de toros Sanmartinero y Cebú. Métodos: se colectó semen de 10 toros de cada raza, y el plasma seminal se sometió a electroforesis bidimensional, para establecer la relación entre la cantidad relativa de cada punto de proteína y la viabilidad espermática. Identificados dichos puntos, el plasma seminal se sometió a cromatografía de exclusión para separar la fracción que contenía estas proteínas. Esta se adicionó en dosis de 0,5, 1,0, 1,5 y 2,0 mg, a muestras de 1 x 10(6) espermatozoides, descongelados e incubados a 37 °C durante 1 hora, para determinar su efecto en la viabilidad post-descongelación. Los espermatozoides se congelaron usando dos medios (citrato-fructosa-yema y Bioxcell®). Resultados: se encontró un punto de proteína (16,20 kDa, punto Isoeléctrico 5,5) en plasma de toros Sanmartinero, que correlacionó (r = 0,64 p<0,001) con la viabilidad. Este punto de proteína se encontró en la fracción 21-25 de la cromatografía. El porcentaje de espermatozoides viables post-descongelación aumentó 20% (p<0,05) con dosis de 1 y 1,5 mg de la fracción, cuando los espermatozoides se congelaron en medio citrato-fructosa-yema; y 25% (p<0,01) con dosis de 0,5 mg cuando se congelaron en medio Bioxcell®. La adición de 0,5 mg de la fracción a semen descongelado previamente criopreservado en medio Bioxcell®, evidenció un incremento mayor (p<0,05) en el porcentaje de espermatozoides viables de semen de toros Sanmartinero (23 ± 8,3 %), que en semen de toros Cebú (6,0 ± 2,0%). Conclusiones: los resultados anteriores demuestran que las proteínas del plasma seminal disminuyen el daño en los espermatozoides por la criopreservación, y que el efecto de estas proteínas depende del medio de congelación, la dosis adicionada y la raza de los toros

Las proteínas del plasma seminal incrementan la viabilidad espermática post-descongelación del semen de toros Sanmartinero

This study was performed to evaluate the effect of the addition of proteins on the postthawing viability of spermatozoa. Materials and methods. Spermatozoa were frozen with two different media: Citrate-fructose and Bioxcell. The isolation of seminal plasma proteins of low molecular weight was performed through low pressure liquid chromatography. It was determined that the proteins of interest eluted in fractions 21-25, and two dimensional electrophoresis was performed. Thawed sperm was incubated at 37°C for one hour with 0.5, 1, 1.5 and 2.0 mg of 21-25 fraction protein. Two additional treatments were included: one with seminal plasma total protein, and another one without protein. Results. Two dimensional electrophoresis of protein confirmed the presence of two bands of 14 and 16 kDa and seven spots with iso-electric points between 5.0 - 5.5 respectively. Incubation of the spermatozoa with the 21-25 fraction showed that sperm viability increases by 20% with doses of 1 and 1.5 mg of protein/106 spermatozoa in the citrate-fructose medium, and 25% with 0.5 mg of protein/106 spermatozoa in Bioxcell medium. A positive effect in sperm viability was demonstrated although it depends on the doses of protein and the cryopreservation medium used. Conclusions. This investigation suggests that the use of seminal plasma proteins can be useful for reducing the harmful effect on sperm cryopreservation.Objetivo. El objetivo de este trabajo fue evaluar el efecto de la adición de proteínas del plasma seminal sobre el porcentaje de espermatozoides bovinos viables post-descongelación. Materiales y métodos. Los espermatozoides se congelaron usando dos medios (citrato-fructosa-yema y Bioxcell) y la obtención de proteínas de plasma seminal de bajo peso molecular se realizó por medio de cromatografía líquida de baja presión. Las proteínas de interés eluyeron en las fracciones 21-25 y se sometieron a electroforésis en una y dos dimensiones. Los espermatozoides se incubaron a 37°C durante una hora, con 0.5, 1.0, 1.5 y 2.0 mg de la fracción 21-25. Se incluyeron dos tratamientos adicionales: uno con proteínas totales del plasma seminal y otro sin proteína. Resultados. La electroforésis bidimensional de las fracciones confirmó la presencia de siete puntos de proteína de bajo peso molecular (14-16 kDa y punto Isoeléctrico de 5.0 - 5.5). La adición de estas proteínas aumentó 20% (p<0.05), el porcentaje de espermatozoides viables post-descongelación en muestras congeladas en medio citrato-fructosa-yema (con dosis de 1 ó 1.5 mg de proteína/106 espermatozoides), y 25% (p<0.05) en muestras congeladas en medio Bioxcell (con dosis de 0.5 mg de proteína/106 espermatozoides). Conclusiones. Los resultados de esta investigación sugieren el posible uso de proteínas de bajo peso molecular del plasma seminal, para disminuir el efecto deletéreo de la criopreservación en los espermatozoides

Survey of gastrointestinal parasites, liver flukes and lungworm in feces from dairy cattle in the high tropics of Antioquia, Colombia

A cross sectional study was undertaken to determine the prevalence and intensity of parasitic infections in dairy cattle in the high tropics of Colombia. A total of 1003 rectal samples were collected from dairy cows at 29 farms between May and June 2014 to represent the number of farms, age groups, and size of the 65,000-cow population in the municipality of San Pedro de los Milagros. Coprological techniques were used to detect gastrointestinal nematodes, liver flukes, coccidian oocysts, and first larval stage counts of Dictyocaulus viviparus. In order of decreasing prevalence, the following parasites were detected: coccidial oocyst (36.7%; 95% CIs, 31.6–42.7), strongyle nematodes (31.6%, 27.8–35.4), liver flukes (30.9%, 21.5–37.5), cestodes (8.4%, 7.1–9.7), and D. viviparus (5.4%, 3.4–7.5). Co-infections by all possible combinations of the three most predominant groups occurred in 11 to 15% of the animals. There were significant differences in infection rates between age groups, with higher risk of liver fluke infection in animals older than 1 year of age (odds ratio (OR) = 3.2), but lower presence for coccidia and strongyles (OR = 0.19 and 0.51, respectively). For Fasciola hepatica, within-herd prevalences of >25% in 16 farms and 94 of 281 (33.5%) animals with >5 eggs per gram (epg) indicate that significant production losses are likely occurring. The variation in the prevalence of gastrointestinal parasites and liver flukes, together with the level of infection among age groups, could be used in integrated management programs to establish selective anthelmintic treatments and select for heritable traits of host resistance. These results serve as a baseline for future studies to determine the success of control measures and should increase awareness that subclinical parasitism is widespread in the livestock sector

Characteristics of household contacts (HHCs) of pulmonary tuberculosis cases and individuals from their source population (SP).

<p>*Only index cases that lead to household contacts included.</p><p>†p value (two tailed) refers to comparison of characteristics in HHCs and individuals from SP.</p><p>‡SES categories as defined in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0008257#s2" target="_blank">Methods</a>.</p

Correlation between TST and IFNγ production in response to CFP and CFP-10 by age in household contacts (HHCs) and individuals from source population (SP).

<p>*Spearman correlation coefficient <i>r</i>. All p values<0.001.</p><p>†TST (mm of induration); IFNγ (pg/mL).</p