Heparin Induces Apoptosis in Lymphocytes from B-cell Chronic Lymphocytic Leukemia.

It has been shown that glycosaminoglycans play a role in the regulation of immune response. In particular, heparin exerts an antiproliferative and apoptotic action in different cellular systems. In this study we evaluate whether heparin can also induce a naturally occurring programmed cell death in lymphocytes from B-chronic lymphocytic leukemia (B-CLL), a neoplastic lineage where apoptosis is blocked by the expression of the proto-oncogene bc1-2. Peripheral blood lymphocytes (PBL) from 7 cases of B-CLL patients in different stages were cultured with three different heparin sodium concentrations for 4 days. Apoptosis was evaluated by agarose gel electrophoresis and by cytofluorimetric analysis. Bcl-2 expression was tested by flow cytometric analysis and immunohistochemistry on cytospin preparations. Agarose gel electrophoresis showed the characteristic DNA fragmentation pattern of apoptosis in all the cases of B-CLL stage III and IV after heparin incubation. DNA from normal and neoplastic lymphocytes cultured without heparin did not undergo spontaneous apoptosis. Cytofluorimetric analysis confirmed the agarose gel pattern and found a level of apoptosis over 50% after culture of neoplastic PBL with heparin. In these cases bcl-2 expression was found to be significantly reduced after heparin incubation when comparing to bcl-2 level before incubation. Our data adds further evidence regarding the potential role of heparin in oncogene inhibition and in apoptosis induction. In particular, the induction of apoptosis in neoplastic lymphocytes by heparin may have a role in the complicated field of interactions between the immune system and the blood vessels by glycosaminoglycans

Detection of nucleic acid-nuclear hormone receptor complexes with mass spectrometry

Nuclear receptors, such as the retinoic acid receptor (RAR) or the 9-cis retinoic acid receptor (RXR), interact not only with their ligands but also with other types of receptors and with DNA. Here, two complementary mass spectrometry (MS) methods were used to study the interactions between retinoic receptors (RXR/RAR) and DNA: non-denaturing nano-electrospray (nanoESI MS), and high-mass matrix-assisted laser desorption ionization (MALDI MS) combined with chemical cross-linking. The RAR·RXR heterodimer was studied in the presence of a specific DNA sequence (DR5), and a specific RAR·RXR·DNA complex was detected with both MS techniques. RAR by itself showed no significant homodimerization. A complex between RAR and the double stranded DR5 was detected with nanoESI. After cross-linking, high-mass MALDI mass spectra showed that the RAR binds the single stranded DR5, and the RAR dimer binds both single and double stranded DR5. Moreover, the MALDI mass spectrum shows a larger RAR dimer signal in the presence of DNA. These results suggest that a gene-regulatory site on DNA can induce quaternary structural changes in a transcription factor such as RA

Sequence-specific transcription factor NF-Y displays histone-like DNA binding and H2B-like ubiquitination

SummaryThe sequence-specific transcription factor NF-Y binds the CCAAT box, one of the sequence elements most frequently found in eukaryotic promoters. NF-Y is composed of the NF-YA and NF-YB/NF-YC subunits, the latter two hosting histone-fold domains (HFDs). The crystal structure of NF-Y bound to a 25 bp CCAAT oligonucleotide shows that the HFD dimer binds to the DNA sugar-phosphate backbone, mimicking the nucleosome H2A/H2B-DNA assembly. NF-YA both binds to NF-YB/NF-YC and inserts an α helix deeply into the DNA minor groove, providing sequence-specific contacts to the CCAAT box. Structural considerations and mutational data indicate that NF-YB ubiquitination at Lys138 precedes and is equivalent to H2B Lys120 monoubiquitination, important in transcriptional activation. Thus, NF-Y is a sequence-specific transcription factor with nucleosome-like properties of nonspecific DNA binding and helps establish permissive chromatin modifications at CCAAT promoters. Our findings suggest that other HFD-containing proteins may function in similar ways

Ante-mortem and Post-mortem Inspection and Relationship between Findings in a North Albanian Pig Slaughterhouse

Simple Summary In European Union abattoirs, the safety of meat is dependent on the favorable opinion from an official veterinarian, in accordance with the current legislation. From this perspective, the feedback generated from the ante-mortem visit and the post-mortem inspection can be investigated to control the health and welfare conditions of the animals in the pre-slaughter phases. From this perspective, we evaluated the ante-mortem and post-mortem inspection outcomes of slaughtered pigs in northern Albania and correlated the results in order to gain insight into the conditions and injuries of pigs slaughtered outside the European context and to extend knowledge on the possible relationship between ante-mortem and post-mortem relief. Dyspnea and tail, skin, and ear lesions were the most frequently observed conditions before slaughter, while pleuritis, pneumonia, liver alterations, white spots on the liver, and pericarditis were the most frequent lesions after slaughter. A significant increase in the total number of post-mortem findings was also observed as the number of ante-mortem findings increased. Overall, the prevalence of the findings observed in this study falls within the broad range of the data in the literature, but additional information should be collected during meat inspection so as to better understand the relationship between ante- and post-mortem outcomes. In June 2014, Albania was granted EU candidate status, thus starting a process of compliance with the membership criteria. In this context, a modern meat inspection approach in line with the European legislation was applied to a pig slaughterhouse in northern Albania in order to investigate the ante-mortem (AM) and post-mortem (PM) conditions and the relationship between these findings. For this purpose, 3930 pigs divided into 35 batches were evaluated over a 3-month period. The most frequent AM conditions recorded were tail lesions and dyspnea (9.1%), followed by skin (8.9%) and ear lesions (8.5%), while in the PM inspections, pleuritis was the most frequently observed condition (10.2%), followed by pneumonia (8.5%), liver alterations (5.7%), milk spot liver (3.8%), and pericarditis (3.3%). With the exception of liver alterations, the other PM lesions mentioned were positively associated with lesions on the ears (OR = 1.036; p < 0.001) and skin (OR = 1.026; p = 0.011) and dyspnea (OR = 1.021; p = 0.005), confirming the link between these variables and the health and welfare conditions of pigs on farms. Overall, the evidence that emerged from this Albanian slaughterhouse can be considered in line with other European contexts, especially in light of the considerable variability in the data present in the literature

Promoter methylation of tumor suppressor genes in pre-neoplastic lesions; Potential marker of disease recurrence

Background: Epigenetic alterations of specific genes have been reported to be related to colorectal cancer (CRC) transformation and would also appear to be involved in the early stages of colorectal carcinogenesis. Little data are available on the role of these alterations in determining a different risk of colorectal lesion recurrence. The aim of the present study was to verify whether epigenetic alterations present in pre-neoplastic colorectal lesions detected by colonoscopy can predict disease recurrence. Methods. A retrospective series of 78 adenomas were collected and classified as low (35) or high-risk (43) for recurrence according to National Comprehensive Cancer Network guidelines. Methylation alterations were analyzed by the methylation-specific multiplex ligation probe assay (MS-MLPA) which is capable of quantifying methylation levels simultaneously in 24 different gene promoters. MS-MLPA results were confirmed by pyrosequencing and immunohistochemistry. Results: Higher levels of methylation were associated with disease recurrence. In particular, MLH1, ATM and FHIT gene promoters were found to be significantly hypermethylated in recurring adenomas. Unconditional logistic regression analysis used to evaluate the relative risk (RR) of recurrence showed that FHIT and MLH1 were independent variables with an RR of 35.30 (95% CI 4.15-300.06, P = 0.001) and 17.68 (95% CI 1.91-163.54, P = 0.011), respectively. Conclusions: Histopathological classification does not permit an accurate evaluation of the risk of recurrence of colorectal lesions. Conversely, results from our methylation analysis suggest that a classification based on molecular parameters could help to define the mechanisms involved in carcinogenesis and prove an effective method for identifying patients at high risk of recurrence

Improved stool DNA integrity method for early colorectal cancer diagnosis

Background: DNA integrity analysis could represent an alternative approach to the early detection of colorectal cancer. Previously, fluorescence long DNA (FL-DNA) in stools was extracted using a manual approach and analyzed by capillary electrophoresis assay (CE FL-DNA). We aimed to improve diagnostic accuracy using a simpler and more standardized method [Real Time PCR FL-DNA (RT FL-DNA)] for the detection of early malignant lesions in a population undergoing colorectal cancer screening. Methods: From 241 stool samples,DNAwas extracted using manual and semiautomatic extraction systems and analyzed using FL-DNA tests by CE and RT assays. The RT FL-DNA approach showed slightly higher sensitivity and specificity compared with the CEFL-DNA method. Furthermore, we compared the RTFL-DNA approach with the iFOBT report. Results: Nonparametric ranking statistics were used to analyze the relationship between the median values of RT FL-DNA and the clinicohistopathologic characteristics. The median values of both variables were significantly higher in patients with cancer than in patients with noncancerous lesions. According to the Fagan nomogram results, the iFOBT and FL-DNA methods provided more accurate diagnostic information and were able to identify subgroups at varying risks of cancer. Conclusions: The combination of the semiautomatic extraction system and RT FL-DNA analysis improved the quality of DNA extracted from stool samples. Impact: RT FL-DNAshows great potential for colorectal cancer diagnosis as it is a reliable and relatively easy analysis to perform on routinely processed stool samples in combination with iFOBT

Meat quality and sensory traits in rabbits fed with two different percentages of bovine colostrum

The nutritional, antimicrobial, and antioxidant properties of bovine colostrum (BC) have encouraged its use in animal nutrition as a functional food in recent years. Nonetheless, the potential implications of BC supplementation on meat quality remain to be thoroughly assessed. To address this, thirty-nine New Zealand White rabbits (n = 13/group) were fed different dietary regimens until slaughter.: commercial standard diet for the control group (C) and C with 2.5% and 5% w/w of BC for BC-2.5 and BC-5 groups, respectively. Rabbits were slaughtered at 91 days of age and meat quality, and sensory characteristics were evaluated at days 2 (48 h after slaughter), 5, and 10 of refrigerated storage at 4 °C. The addition of colostrum in the diet resulted in a reduction of the total viable count, albeit only at the highest concentration and at the final detection, whereas for Lactobacillus spp. and Pseudomonas spp., there was little or no effect. The colour coordinates showed no differences between the groups, but they varied over time according to diet. Some differences between groups emerged in the definition of sensory attributes but did not affect the overall liking and overall scores of individual descriptors. These results indicate that the use of colostrum in rabbit feeding does not significantly impart meat quality and sensory attributes, but the potential of this valuable by-product for the food industry needs further investigation

Ideología Política de dirigentes de la Ciudad de Juliaca

La investigación tuvo como objetivo determinar el tipo de ideología política de dirigentes de la ciudad de Juliaca, el problema que orientó esta investigación fue el liderazgo representado a través de ideología política. Se usó una metodología no experimental y el diseño descriptivo de tipo básico. El estudio estuvo conformado por una muestra de 92 dirigentes de urbanizaciones de la ciudad de Juliaca, que se obtuvo de forma aleatoria simple. Se empleó para esta investigación una técnica de evaluación objetiva, cuyo instrumento fue el Test de Nolan, adaptado al contexto y validado de acuerdo a las pruebas piloto que se realizaron. La situación económica, social y política que se estudió, refiere que Juliaca tiene un carácter heterogéneo en sus ciudadanos, porque alberga diferentes grupos sociales que provienen de distintos distritos aledaños a Juliaca, principalmente por el comercio que se lleva a cabo en esta ciudad. Los resultados, en base a los objetivos planteados, determinaron que existe una ideología predominante en los dirigentes de Juliaca, la centralista (44,6 %), seguida de la totalitaria (32,6 %), lo que significa que comparten características con otras ideologías, prefiriendo una combinación equilibrada entre Estado y libertad individual. Situación que refiere una característica predominante en los dirigentes de la ciudad de Juliaca, de acuerdo al estudio realizado, sería la práctica del clientelismo político. Finalmente, se indica que los dirigentes de diferentes grupos sociales de la ciudad de Juliaca tienen una ideología política centro y totalitario Palabras clave: Ideología, política, dirigente