Producción de embriones de búfalo por fertilización in vitro luego de la maduración de los ovocitos durante el transporte prolongado

El búfalo (Bubalus bubalis) es una especie con excelente adaptación a sectores inundables. El mejoramiento genético a través de superovulación y transferencia embrionaria ha tenido escasos resultados debido a dificultades en la detección de celo, pobre respuesta ovárica y limitada recuperación de embriones post-lavaje. La técnica de fertilización in vitro de embriones (FIV) es una biotecnología de gran impacto en el progreso genético. El objetivo del presente trabajo fue estudiar los eventos tempranos de la FIV, analizando la tasa de maduración y desarrollo embrionario post-fertilización de ovocitos madurados in vitro (IVM) durante el transporte. Ovocitos bovinos y bubalinos fueron obtenidos por punción folicular de ovarios post-mortem e IVM durante el transporte por un período de 18 h. Se realizó la FIV con toros de fertilidad comprobada, con una concentración en microgotas de inseminación de 3-4 x 106 espermatozoides motiles/ml por un período de 6 horas. Los embriones fueron cultivados en medio oviductal sintético SOFaa en incubadora gaseada y ambiente humidificado a 38,5ºC durante 9 días. Se evaluaron las tasas de IVM, clivaje (día 2 post-fertilización) y blastocisto (días 7 a 9). Los resultados fueron analizados estadísticamente utilizando Fischer’s Exact Test (

Primary clear cell sarcoma of the ileum: an uncommon and misleading site

A clear cell sarcoma, arising primarily in the ileum of a 35-year-old man, is reported. Histologically, the neoplasm infiltrated the full thickness of the intestinal wall. It consisted of strands and sheets of round to spindle-shaped cells with clear to eosinophilic cytoplasm, vesicular nuclei and prominent nucleoli. Vascular invasion was present at diagnosis. Tumour cells expressed S-100 protein, melan-A and tyrosinase. They were negative for HMB45, CD117, cytokeratins, epithelial membrane antigen, smooth muscle actin, desmin, CD31, CD34, chromogranin and synaptophysin. Reverse transcription-polymerase chain reaction analysis performed on paraffin-embedded tissue showed EWS-ATF1 fusion transcripts representative of the t(12;22) (q13;q12) clear cell sarcoma reciprocal translocation. The patient, who developed liver metastases 2 months after diagnosis, died of disease at 15 months. This case demonstrates that the gastrointestinal tract is a potential site for primary clear cell sarcoma of soft tissues, and, furthermore, that cytogenetics and/or molecular techniques play a central role in the diagnosi

[Experimental infection of calves and sheep with bovine Giardia isolates].

9 Giardia-free calves were artificially infected with 1.5-5.1 x 10(6) Giardia cysts originating from Swiss cattle ("bovine isolates"). In 4 of these animals the course of infection was examined. After prepatent periods of 7-8 days all calves excreted high numbers of Giardia cysts for 60-112 days. During patency on 44% of the examination days Giardia cysts and antigen could be detected simultaneously in faecal samples using the flotation method and a sandwich-ELISA, respectively. With the exception of light diarrhoea lasting only for some days at the beginning of patency no other symptoms occurred. Further 5 artificially infected calves were submitted to autopsy. Giardia trophozoites were detected in 4 calves in the jejunum and in 1 animal in the ileum (peroxidase-antiperoxidase method). All animals were simultaneously infected with Campylobacter spp. and/or Rota- and Corona-virus. Electronmicroscopically mucosal attachment sites of Giardia trophozoites had intact microvilli and enterocytes. In various parts of the intestine blunting and flattening of the villi and cellular infiltrations of the mucosa were present. These alterations in calves are generally associated with bacterial and/or viral infections of calves. A Swiss bovine Giardia cyst-isolate was transmitted to 4 Giardia-free conventionally maintained lambs which excreted Giardia cysts after prepatent periods of 10-21 days for 31-61 days

A cost-effective algorithm for hereditary nonpolyposis colorectal cancer detection.

Colorectal cancer with microsatellite instability (MSI) may occur sporadically or be inherited in cases of hereditary nonpolyposis colorectal cancer (HNPCC) syndrome. However, there is no consensus as to which patients must be tested and how to test MSI. In this study, MSI was tested by immunohistochemical analysis and by polymerase chain reaction in 148 cases of colorectal cancer, and methylation of the hMLH1 promoter was examined. MSI status was correlated with tumor phenotype. We found that localization, tumor infiltrating lymphocytes, and mucinous differentiation were predictive of high-frequency MSI (MSI-H) colorectal cancer and might be used to select cases for MSI analysis. Immunohistochemical analysis detected most MSI-H colorectal cancer and might constitute the first step in MSI detection. Absence of hMLH1 promoter methylation in MSI-H colorectal cancer could be predictive of hereditary colorectal cancer, and, hence, methylation analysis might constitute the second step in the identification of patients with HNPCC

Increasing Incidence of Microscopic Colitis in a Population-Based Cohort Study in Switzerland.

Microscopic colitis (MC) is a chronic inflammatory disease of the colon that presents with chronic, nonbloody watery diarrhea and only few or no endoscopic abnormalities. Histologic examination discriminates lymphocytic colitis (LyC; presence of ≥20 intraepithelial lymphocytes per 100 surface epithelial cells) and collagenous colitis (CC; colonic subepithelial collagen band >10 μm in diameter). <sup>1</sup> <sup>,</sup> <sup>2</sup> MC not otherwise specified describes a subgroup of patients who do not fulfill the diagnostic criteria for either CC or LyC. <sup>1</sup> <sup>,</sup> <sup>2</sup> Population-based epidemiologic data regarding MC are scarce. We aimed to evaluate the clinical presentation at diagnosis, incidence, and prevalence of MC in Cantons of Vaud and Fribourg, Switzerland

APC senses cell-cell contacts and moves to the nucleus upon their disruption

Fil: Brocardo, Mariana G. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Bianchini, Michele. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Radrizzani, Martín. ANLIS Dr.C.G.Malbrán. Centro Nacional de Genética Médica; Argentina.Fil: Reyes, Gloria B. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Dugour, Andrea V. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Taminelli, Guillermo L. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Gonzalez Solveyra, César. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.Fil: Santa-Coloma, Tomás A. Instituto de Investigaciones Bioquímicas. Fundación Campomar; Argentina.The adenomatous polyposis coli (APC) tumor suppressor protein is involved in the Wnt/wingless pathway, modulating beta-catenin activity. We report the development of a highly specific, chemically synthesized oligobody (oligonucleotide-based synthetic antibody), directed against the N-terminal region of APC. Using this reagent, we found that within 16 h of disrupting HT-29 cell-cell contacts by harvesting cells with trypsin/EDTA treatment and replating, APC was translocated from the cytoplasm to the nucleus. Five days after plating the cells, when the cells had returned to their normal confluent phenotype and cell-cell contacts were reestablished, APC returned to the cytoplasm. These results suggest that APC functions as part of a "sensor" system, and responds to the loss of cell-cell contacts by moving to the nucleus, and returning to the cytoplasm when the contacts are fully restored

Mitochondrial complex I in-gel activity (IGA) measured in cells expressing wt-CFTR.

<p>A: IGA of mitochondrial extracts from T84 cells after 24 h of treatment with 100 µM glibenclamide or 5 µM CFTR(inh)-172. B: Densitometric quantification of the results shown in panel A, expressed as % ratio of (mCx-I activity)/(protein load). C: IGA of mitochondrial extracts from T84 cells after 48 h of treatment with 100 µM glibenclamide or 5 µM CFTR(inh)-172. D: Densitometric queantification of C. E: IGA of the mCx-I from Caco-2 cells after 48 h of treatment with 5 µM GlyH101 or 5 µM CFTR(inh)-172, and WB of the mCx-III subunit UQCRC1, as internal standard. F: Densitometric quantification of the results shown in E expressed as % ratio of (mCx-I IGA)/UQCRC1(a.u.). The activity of mCx-I in T84 and Caco-2 cells treated with the same amount of DMSO (0.1%) was considered as 100%. Measurements in T84 cells were performed in duplicate and data are expressed as mean ± SE of three independent experiments (n = 3). Caco-2 cells results were obtained in triplicate and expressed as mean ± SE of three independent experiments (n = 3). * indicates p<0.05, as compared with control cells (ANOVA and Turkey's test).</p

Mitochondrial complex I in-gel activity (IGA) of IB3-1 and S9 cells.

<p>A: IGA of mitochondrial extracts from Control and CFTR-stimulated IB3-1 and S9 cells (IBMX-isop-cAMP), adding 200 µM cAMP, 10 µM isoproterenol, 200 µM IBMX, for 24 h. B: Densitometric quantification and statistical analysis of the results shown in panel A. IGA was calculated as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048059#pone-0048059-g001" target="_blank">Figure 1</a>. Measurements were performed in duplicate and data are expressed as mean ± SE of five independent experiments (n = 5). * indicates p<0.05, as compared with S9 stimulated cells. C: IGA of mCx-I and mCx-III (UQCRC1) expression measured by using Western blots from S9 and IB3-1 cells (both after CFTR stimulation). D: Densitometric quantification and statistical analysis of the results shown in panel C. IGA of mCx-I was calculated as the ratio mCx-I IGA/UQCRC1. Measurements were performed in duplicate and data are expressed as mean ± SE of two independent experiments (n = 2). * indicates p<0.05, as compared with S9 stimulated cells. E: IGA of the mCx-I and Coomassie blue stain from a BN-PAGE using mitochondrial extracts from S9 and IB3-1 cells. F: Specific activity of the results shown in panel E, calculated as the ratio mCx-I IGA/mCx-I coomassie blue stain. The mCx-I specific activity is expressed in arbitrary units (a.u.) as mean ± SE (n = 3). The specific activity of CF cells (IB3-1) and CF corrected cells (S9) showed similar values, without significant differences (p>0.05).</p