Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas

Esta publicación detalla los experimentos realizados para la clonación de un ADNc que codifica una nueva metaloproteasa de matriz extracelular a partir de una biblioteca de ADNc procedente de un carcinoma mamario. Este trabajo es de gran interés en la investigación del cáncer, ya que describe la identificación de una nueva colagenasa en los carcinomas mamarios proponiendo un posible papel en el proceso tumoral. Hay evidencia de que las metaloproteasas participan en el proceso de degradación proteolítica de los diferentes componentes de la membrana basal, favoreciendo así la invasión tumoral y las metástasis. El ADNc de la colagenasa-3 se expresó en un sistema de virus vaccinia, y la proteína recombinante fue capaz de degradar los colágenos fibrilares, lo que apoya la hipótesis de que el ADNc aislado codifica para una colagenasa auténtica. El análisis por Northern blot del ARN de tejidos normales y patológicos demostró la existencia de tres especies diferentes de ARNm en los tumores de mama, que parecen ser el resultado de la utilización de distintos sitios de poliadenilación presentes en la región 3'-no codificante del gen. Por el contrario, no se detectó ARNm de la colalagenasa-3 por Northern blot ni por PCR en otros tejidos humanos como mama normal, fibroadenomas mamarios, hígado, placenta, ovario, útero, próstata y glándula parótida. Sobre la base del aumento de la expresión de la colagenasa-3 en los carcinomas de mama y la ausencia de expresión detectable en los tejidos normales, se propone un posible papel de esta metaloproteinasa en el proceso tumoral

Novel insights on intake of fish and prevention of sarcopenia: All reasons for an adequate consumption

Sarcopenia is defined as a syndrome characterized by progressive and generalized loss of skeletal muscle mass and strength and it is diagnosed by measurements of muscle mass, muscle strength, and physical performance. Sarcopenia affects quality of life and is associated with several adverse health effects. Muscle decline is aggravated by a sedentary lifestyle and can be prevented through proper nutrition, together with adequate physical activity. Fish contains biologically active compounds, such as omega-3 polyunsaturated fatty acids, proteins, vitamin D, magnesium, and carnitine, which are able to intervene positively on muscle metabolism. This narrative literature review was performed to evaluate evidence regarding the actual benefit of fish consumption in the prevention of sarcopenia and the positive action on the muscle mass of the biological compounds present in fish. The results demonstrated that fish consumption has a protective and anti-inflammatory function on skeletal muscle and that its biologically active compounds help to maintain good muscle performance, preventing sarcopenia. Considering the nutritional and health benefits, elderly with sarcopenia should consume at least three servings per week of fish in order to have a minimum intake of 4–4.59 g daily of omega 3, and reaching the 50% RDA in Vitamin E and D. High biological value of proteins in 150 g of fish and its high available magnesium (20% of RDA in 150 g of fish) are an added value that could suggest fish as a “functional food” in order to prevent and treat sarcopenia

A family case of fertile human 45,X,psu dic(15;Y) males

We report on a familial case including four male probands from three generations with a 45,X,psu dic(15;Y)(p11.2;q12) karyotype. 45,X is usually associated with a female phenotype and only rarely with maleness, due to translocation of small Y chromosomal fragments to autosomes. These male patients are commonly infertile because of missing azoospermia factor regions from the Y long arm. In our familial case we found a pseudodicentric translocation chromosome, that contains almost the entire chromosomes 15 and Y. The translocation took place in an unknown male ancestor of our probands and has no apparent effect on fertility and phenotype of the carrier. FISH analysis demonstrated the deletion of the pseudoautosomal region 2 (PAR2) from the Y chromosome and the loss of the nucleolus organizing region (NOR) from chromosome 15. The formation of the psu dic(15;Y) chromosome is a reciprocal event to the formation of the satellited Y chromosome (Yqs). Statistically, the formation of 45,X,psu dic(15;Y) (p11.2;q12) is as likely as the formation of Yqs. Nevertheless, it has not been described yet. This can be explained by the dicentricity of this translocation chromosome that usually leads to mitotic instability and meiotic imbalances. A second event, a stable inactivation of one of the two centromeres is obligatory to enable the transmission of the translocation chromosome and thus a stably reduced chromosome number from father to every son in this family

Graphical Performance Measures for Practitioners to Triage Split Failure Trouble Calls

Detector occupancy is commonly used to measure traffic signal performance. Despite improvements in controller computational power, there have been relatively few innovations in occupancy-based performance measures or integration with other data. This paper introduces and demonstrates the use of graphical performance measures based on detector occupancy ratios to verify potential split failures and other signal timing shortcomings reported to practitioners by the public. The proposed performance measures combine detector occupancy during the green phase, detector occupancy during the first five seconds of the red phase, and phase termination cause (gap out or force off). These are summarized by time of day to indicate whether the phase is undersaturated, nearly saturated, or oversaturated. These graphical performance measures and related quantitative summaries provide a first-level screening and triaging tool for practitioners to assess user concerns regarding whether sufficient green times are being provided to avoid split failures. They can also provide outcome-based feedback to staff after making split adjustments to determine whether operation improved or worsened. The paper concludes by demonstrating how the information was used to make an operational decision to re-allocate green time that reduced the number of oversaturated cycles on minor movements from 304 to 222 during a Thursday 0900-1500 timing plan and from 240 to 180 during a Friday 0900-1500 timing plan

Chronic social stress induces peripheral and central immune activation, blunted mesolimbic dopamine function, and reduced reward-directed behaviour in mice

Psychosocial stress is a major risk factor for depression, stress leads to peripheral and central immune activation, immune activation is associated with blunted dopamine (DA) neural function, DA function underlies reward interest, and reduced reward interest is a core symptom of depression. These states might be inter-independent in a complex causal pathway. Whilst animal-model evidence exists for some specific steps in the pathway, there is currently no animal model in which it has been demonstrated that social stress leads to each of these immune, neural and behavioural states. Such a model would provide important existential evidence for the complex pathway and would enable the study of causality and mediating mechanisms at specific steps in the pathway. Therefore, in the present mouse study we investigated for effects of 15-day resident-intruder chronic social stress (CSS) on each of these states. Relative to controls, CSS mice exhibited higher spleen levels of granulocytes, inflammatory monocytes and T helper 17 cells; plasma levels of inducible nitric oxide synthase; and liver expression of genes encoding kynurenine pathway enzymes. CSS led in the ventral tegmental area to higher levels of kynurenine and the microglia markers Iba1 and Cd11b and higher binding activity of DA D1 receptor; and in the nucleus accumbens (NAcc) to higher kynurenine, lower DA turnover and lower c-fos expression. Pharmacological challenge with DA reuptake inhibitor identified attenuation of DA stimulatory effects on locomotor activity and NAcc c-fos expression in CSS mice. In behavioural tests of operant responding for sucrose reward validated as sensitive assays for NAcc DA function, CSS mice exhibited less reward-directed behaviour. Therefore, this mouse study demonstrates that a chronic social stressor leads to changes in each of the immune, neural and behavioural states proposed to mediate between stress and disruption of DA-dependent reward processing. The model can now be applied to investigate causality and, if demonstrated, underlying mechanisms in specific steps of this immune-neural-behavioural pathway, and thereby to identify potential therapeutic targets

Improving Intersection Behavior through Delay-Based Left Turn Phase Initiation

Serving protected left-turn phases for one or two vehicles can often be an inefficient use of cycle green time when the opposing through movements are over capacity. This paper assesses the performance of an intersection at which controller logic is applied to delay the call for a protected left-turn phase on the basis of vehicle wait times. During four weeks of evaluation, the delay on left-turn phase calls was varied in 25-s increments from 0 to 75 s. The results indicate that delaying left-turn phase initiation substantially increases the amount of green time for saturated through movements while minimally increasing the travel delay for left-turning drivers. The recommendation is made for agencies to consider using a delay in the range of 25 to 50 s for calling protected phases at intersections at which the opposing through movement is oversaturated and could benefit from additional green time. This paper presents one of the first quantitative studies evaluating potential intersection capacity and performance improvements with respect to left-turn detector delay. </jats:p

DNA damage triggers squamous metaplasia in human lung and mammary cells via mitotic checkpoints

Epithelial transdifferentiation is frequent in tissue hyperplasia and contributes to disease in various degrees. Squamous metaplasia (SQM) precedes epidermoid lung cancer, an aggressive and frequent malignancy, but it is rare in the epithelium of the mammary gland. The mechanisms leading to SQM in the lung have been very poorly investigated. We have studied this issue on human freshly isolated cells and organoids. Here we show that human lung or mammary cells strikingly undergo SQM with polyploidisation when they are exposed to genotoxic or mitotic drugs, such as Doxorubicin or the cigarette carcinogen DMBA, Nocodazole, Taxol or inhibitors of Aurora-B kinase or Polo-like kinase. To note, the epidermoid response was attenuated when DNA repair was enhanced by Enoxacin or when mitotic checkpoints where abrogated by inhibition of Chk1 and Chk2. The results show that DNA damage has the potential to drive SQM via mitotic checkpoints, thus providing novel molecular candidate targets to tackle lung SCC. Our findings might also explain why SCC is frequent in the lung, but not in the mammary gland and why chemotherapy often causes complicating skin toxicity

GliomaPredict: a clinically useful tool for assigning glioma patients to specific molecular subtypes

<p>Abstract</p> <p>Background</p> <p>Advances in generating genome-wide gene expression data have accelerated the development of molecular-based tumor classification systems. Tools that allow the translation of such molecular classification schemas from research into clinical applications are still missing in the emerging era of personalized medicine.</p> <p>Results</p> <p>We developed GliomaPredict as a computational tool that allows the fast and reliable classification of glioma patients into one of six previously published stratified subtypes based on sets of extensively validated classifiers derived from hundreds of glioma transcriptomic profiles. Our tool utilizes a principle component analysis (PCA)-based approach to generate a visual representation of the analyses, quantifies the confidence of the underlying subtype assessment and presents results as a printable PDF file. GliomaPredict tool is implemented as a plugin application for the widely-used GenePattern framework.</p> <p>Conclusions</p> <p>GliomaPredict provides a user-friendly, clinically applicable novel platform for instantly assigning gene expression-based subtype in patients with gliomas thereby aiding in clinical trial design and therapeutic decision-making. Implemented as a user-friendly diagnostic tool, we expect that in time GliomaPredict, and tools like it, will become routinely used in translational/clinical research and in the clinical care of patients with gliomas.</p

The Degradome database: mammalian proteases and diseases of proteolysis

The degradome is defined as the complete set of proteases present in an organism. The recent availability of whole genomic sequences from multiple organisms has led us to predict the contents of the degradomes of several mammalian species. To ensure the fidelity of these predictions, our methods have included manual curation of individual sequences and, when necessary, direct cloning and sequencing experiments. The results of these studies in human, chimpanzee, mouse and rat have been incorporated into the Degradome database, which can be accessed through a web interface at http://degradome.uniovi.es. The annotations about each individual protease can be retrieved by browsing catalytic classes and families or by searching specific terms. This web site also provides detailed information about genetic diseases of proteolysis, a growing field of great importance for multiple users. Finally, the user can find additional information about protease structures, protease inhibitors, ancillary domains of proteases and differences between mammalian degradomes