Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs

Assessment of gastrointestinal permeability in vivo is considered a suitable method for the evaluation of gastrointestinal mucosal integrity. Probes commonly used include lactulose (L) and rhamnose (R) for the assessment of intestinal permeability, xylose (X) and 3-O-methylglucose (M) for the evaluation of intestinal absorptive capacity, and sucrose (S) for the assessment of gastric permeability. Traditionally, various methods have been used to quantify these markers in the urine after orogastric administration. However, urine collection is difficult and uncomfortable. A protocol based on the analysis of blood samples would be easier to perform. Thus, the aim of the first part of this project was to develop and validate a new gas chromatography-mass spectrometry (GC-MS) method for the quantification of five sugar probes in canine serum. The method was sensitive, accurate, precise, and reproducible for the simultaneous quantification of 5 sugar probes in serum. The aim of the second part of this project was to assess the kinetic profiles of these 5 sugar probes in serum after orogastric administration in dogs and to determine the optimal time point for sample collection. Dogs received a solution containing L (10 g/L), R (10 g/L), X (10 g/L), M (5 g/L), and S (40 g/L) by orogastric intubation. Baseline blood samples were collected. Subsequent timed blood samples were taken for a 24 hours period. Significant changes in serum concentrations of all 5 sugars were detected after administration of the test dose (p<0.0001 for all 5 probes). Serum concentrations of L and R were significantly different from baseline concentrations from 90 to 240 and from 60 to 300 min post dosing respectively, and those of X, M, and S were significantly different from 30 to 240 min after dosing (p<0.05 for all 5 probes). Variations of the mean sugar concentrations of all dogs at 90, 120, and 180 minutes were analyzed using a Kruskal-Wallis test. Based on the results, only two blood samples, one taken at baseline and a second sample obtained between 90 and 180 after dosing, appear to be sufficient for assessment of intestinal permeability and mucosal absorptive capacity using these sugar probes

Ocjena ejakulata: može li se procijeniti plodnost?

This review critically comments on established and emerging methods of semen evaluation that would foresee the fertility of the ejaculate (neat or processed) or the male. Functional sperm testing through biomimetic in vitro assays are priority since these, by resembling events during sperm transport, storage and interaction with the female genital tract and the oocyte, best provide clues for sperm selection and the role of sperm sub-populations in the ejaculate. The review also focuses on the exploration of the genome, the transcriptome and the proteome of both spermatozoa and the seminal plasma which may help to unveil how spermatozoa and seminal plasma components would signal to the female and link to fertility and even prolificacy. Specific seminal plasma components, both among individuals and portions of the ejaculate, not only relate to sperm survival but also to signal the female immune system towards a differential immune tolerance warranting fertility.Ovaj pregledni članak pruža kritički osvrt na standardne i inovativne metode za procjenu kakvoće sjemena kako bi se predvidio oplodni potencijal ejakulata (nativnog ili obrađenog) i mužjaka. Funkcionalna testiranja spermija u in vitro uvjetima su nužna, budući da se slični procesi odvijaju tijekom transporta spermija, pohrane i interakcije sa ženskim spolnim sustavom i oocitama, jer pružaju uvid u ulogu pojedinih subpopulacija spermija u ejakulatu i time pomažu kod odabira spermija. Nadalje, ovaj se članak isto tako usredotočuje na istraživanje genoma, transkriptoma i proteoma spermija i sjemene plazme što može pomoći pri otkrivanju kako spermiji i molekule sjemene plazme mogu dati signal ženki i povezati to s plodnošću pa čak i s brojem potomaka. Specifične molekule sjemene plazme koje se razlikuju između jedinki između pojedinih frakcija ejakulata, povezane su s preživljavanjem spermija kao i s majčinom imunosnom tolerancijom prema spermijima što omogućuje plodnosti

Modulation of Sperm Function during Sperm Transport in the Female

In the cow, sow and mare, a functional tubal sperm reservoir is established before ovulation to ensure availability of suitable numbers of viable spermatozoa for fertilization. Although identification of subpopulations reaching this reservoir has been attempted, it is still unclear whether this recruitment is programmed or fortituous. Those spermatozoa not reaching the oviduct are generally destroyed by phagocytosis of invading leukocytes. While the type of ejaculate differs in these species, seminal plasma proteins and/or the spermatozoa appear to act as leukocyte chemoattractant both in vitro and in vivo. Those spermatozoa in the sperm reservoir not only escape phagocytosis or rejection by the female immune system but sustain viability and potential fertilizing capacity by not capacitating or acrosome-reacting while residing in the oviduct. Sperm numbers in the reservoir diminish gradually in relation to ovulation, spermatozoa being continuously redistributed towards the upper isthmus. In vitro, only uncapacitated spermatozoa bind to epithelial explants, suggesting that the reservoir milieu modulates sperm capacitation. In vivo, most viable spermatozoa during preovulatory spontaneous standing oestrus are uncapacitated, with capacitation significantly increasing after ovulation. In either species, there seems to be different components of the oviductal fluid effecting capacitation, and bicarbonate appears to be common denominator for the membrane destabilizing changes that encompasses the first stages of the process. Such effects can be blocked or even reversed by co-incubation with isthmic fluid or specific glycosaminoglycans such as hyaluronan. Although the pattern of response to in vitro induction of sperm capacitation is similar for all spermatozoa, the capacity of response and its speed is very individual. Such diverse individual response to capacitation would not only confirm capacitation does not occur massively in the reservoir but clearly insures full sperm viability before ovulation and the presence of spermatozoa at different stages of capacitation in the upper oviduct, thus maximizing the chances of normal fertilization

“ANÁLISIS DE LAS POLÍTICAS PÚBLICAS PARA EL CAMPO: ESTUDIO DE CASO EL DESARROLLO RURAL SUSTENTABLE DE LA ALIANZA PARA EL CAMPO 2003-2006”

Con el paso del tiempo las economías tienden a crecer y otras a rezagarse, ¿de qué depende esto? De las políticas económicas, de las políticas de desarrollo, de la situación cultural de los países, o simplemente de una buena ejecución de la administración pública. En países más pequeños el concepto de campesino o rural describe una tipología de sociedad con alto nivel de riqueza, en nuestro país ese concepto está asociado a pobrezas y marginalidad. ¿Qué pasa en México? Que es lo que no se está haciendo bien, sobre todo en el ámbito rural. Este escrito se realizó con el afán de conocer el combate a la pobreza en el sector rural y las estrategias que el gobierno toma ante este problema, partiendo de una visión económica de mayor apertura comercial y políticas basadas en el mercado y de las directrices de los países inmersos en los tratados comerciales internacionales. El presente trabajo describe y analiza el Programa de Desarrollo Rural (PDR) que se desprende de la política pública de Alianza para el Campo, formulado en la década de los 90s, detonado por la apertura comercial que el gobierno neoliberal adopta para poder incursionar en el grupo de los países subdesarrollados. Mostrando como el PDR forma parte de las estrategias del gobierno para disminuir el rezago económico en las zonas rurales, el trabajo también muestra mediante el análisis cómo el gobierno apoya la inversión productiva en la agricultura, ganadería y actividades no agrícolas impulsando la demanda de proyectos específicos, con atención a grupos prioritarios, mujeres, indígenas, jóvenes, y discapacitados. Aunado a las cuestiones políticas y sociales es necesario también describir el trabajo legal que sustenta a esta política pública, por ello también, el presente trabajo, describe la evolución del enfoque de México en política rural y el nuevo marco institucional establecido por la Ley de Desarrollo Rural Sustentable desde 2003. Revisa en general las políticas sectoriales existentes que influye en el desarrollo rural, analizando los programas correspondientes, en particular la política integral para el desarrollo rural: el Programa de Desarrollo Rural de la Secretaría de Agricultura, Ganadería, Desarrollo Rural, Pesca y Alimentación (SAGARPA)

Advances in Boar Semen Cryopreservation

The present paper highlights aspects of the cryopreservation of boar semen, a species with particular large, fractionated ejaculates, and a cumbersome cryotechnology that had prevented its commercial application. With the dramatic increase of use of liquid pig semen for artificial breeding over the past decade, developments on cryopreservation alongside the routine use of stud boar semen for AI had been promoted. Recent advances in our laboratory, accommodating the best use of portions of the sperm-rich fraction of the ejaculate for cryopreservation of the sperm-peak portion (P1) and parallel use of the rest of the collected ejaculated spermatozoa, appears as a suitable commercial alternative

Ocjena ejakulata: može li se procijeniti plodnost?

This review critically comments on established and emerging methods of semen evaluation that would foresee the fertility of the ejaculate (neat or processed) or the male. Functional sperm testing through biomimetic in vitro assays are priority since these, by resembling events during sperm transport, storage and interaction with the female genital tract and the oocyte, best provide clues for sperm selection and the role of sperm sub-populations in the ejaculate. The review also focuses on the exploration of the genome, the transcriptome and the proteome of both spermatozoa and the seminal plasma which may help to unveil how spermatozoa and seminal plasma components would signal to the female and link to fertility and even prolificacy. Specific seminal plasma components, both among individuals and portions of the ejaculate, not only relate to sperm survival but also to signal the female immune system towards a differential immune tolerance warranting fertility.Ovaj pregledni članak pruža kritički osvrt na standardne i inovativne metode za procjenu kakvoće sjemena kako bi se predvidio oplodni potencijal ejakulata (nativnog ili obrađenog) i mužjaka. Funkcionalna testiranja spermija u in vitro uvjetima su nužna, budući da se slični procesi odvijaju tijekom transporta spermija, pohrane i interakcije sa ženskim spolnim sustavom i oocitama, jer pružaju uvid u ulogu pojedinih subpopulacija spermija u ejakulatu i time pomažu kod odabira spermija. Nadalje, ovaj se članak isto tako usredotočuje na istraživanje genoma, transkriptoma i proteoma spermija i sjemene plazme što može pomoći pri otkrivanju kako spermiji i molekule sjemene plazme mogu dati signal ženki i povezati to s plodnošću pa čak i s brojem potomaka. Specifične molekule sjemene plazme koje se razlikuju između jedinki između pojedinih frakcija ejakulata, povezane su s preživljavanjem spermija kao i s majčinom imunosnom tolerancijom prema spermijima što omogućuje plodnosti

Development and analytical validation of a gas chromatography-mass spectrometry method for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs

Assessment of gastrointestinal permeability in vivo is considered a suitable method for the evaluation of gastrointestinal mucosal integrity. Probes commonly used include lactulose (L) and rhamnose (R) for the assessment of intestinal permeability, xylose (X) and 3-O-methylglucose (M) for the evaluation of intestinal absorptive capacity, and sucrose (S) for the assessment of gastric permeability. Traditionally, various methods have been used to quantify these markers in the urine after orogastric administration. However, urine collection is difficult and uncomfortable. A protocol based on the analysis of blood samples would be easier to perform. Thus, the aim of the first part of this project was to develop and validate a new gas chromatography-mass spectrometry (GC-MS) method for the quantification of five sugar probes in canine serum. The method was sensitive, accurate, precise, and reproducible for the simultaneous quantification of 5 sugar probes in serum. The aim of the second part of this project was to assess the kinetic profiles of these 5 sugar probes in serum after orogastric administration in dogs and to determine the optimal time point for sample collection. Dogs received a solution containing L (10 g/L), R (10 g/L), X (10 g/L), M (5 g/L), and S (40 g/L) by orogastric intubation. Baseline blood samples were collected. Subsequent timed blood samples were taken for a 24 hours period. Significant changes in serum concentrations of all 5 sugars were detected after administration of the test dose (p<0.0001 for all 5 probes). Serum concentrations of L and R were significantly different from baseline concentrations from 90 to 240 and from 60 to 300 min post dosing respectively, and those of X, M, and S were significantly different from 30 to 240 min after dosing (p<0.05 for all 5 probes). Variations of the mean sugar concentrations of all dogs at 90, 120, and 180 minutes were analyzed using a Kruskal-Wallis test. Based on the results, only two blood samples, one taken at baseline and a second sample obtained between 90 and 180 after dosing, appear to be sufficient for assessment of intestinal permeability and mucosal absorptive capacity using these sugar probes

INCIDENCIA DE LA POSESION PACIFICA DE LOS TERRENOS AGRICOLAS EN EL DERECHO A LA PRESCRIPCION ADQUISITIVA DE DOMINIO DE LOS POBLADORES DE LA PROVINCIA DE PUERTO INCA 2017

Mi tesis tuvo como finalidad especial establecer claramente la incidencia de la posesión pacífica de los terrenos agrícolas en el derecho a la prescripción adquisitiva de dominio de los pobladores de la Provincia de Puerto Inca 2017. Fue plantear la incidencia de la posesión pacífica de los terrenos agrícolas en el derecho a la prescripción adquisitiva de dominio de los pobladores de la Provincia de Puerto Inca 2017, El artículo 950° del Código Civil prescribe que “La propiedad inmueble se adquiere por prescripción mediante la posesión continua, pacífica y pública como propietario durante diez años. Se adquiere a los cinco años cuando median justo título y buena fe.” Debemos indicar que tenemos tres tipos de prescripción adquisitiva de dominio, Vía Administrativa Predios Rurales según Ley N° 27867 Art. 51°; Vía Notarial Predios Urbanos con o sin edificación Ley N° 27157 Art. 21° y Vía Judicial Predios Rústicos y Urbanos Art. 486° y 504° del C.P.C.. En tal sentido, se puede afirmar que la posesión pacífica debe ser entendida como aquella que se ejerce sin perturbación y aceptación de los demás integrantes de la sociedad. Simplemente he querido plantear el tema del fundamento de la incidencia de la posesión pacifica porque creo que ofrece un enorme espacio para la reflexión. Lo que está expresado aquí, está sujeto a crítica y si eso ocurre se habrá cumplido mi propósito, porque nadie es absoluto, lo cual permitirá a los pobladores, más aún a todos los agricultores de la Provincia de Puerto Inca plantear su demanda para la Prescripción Adquisitiva de dominio de sus Inmuebles donde se encuentran posesionados.Tesi