Spontaneous activity in peripheral diaphragmatic lymphatic loops.

The spontaneous contractility of FITC-dextran-filled lymphatics at the periphery of the pleural diaphragm was documented for the first time “in vivo” in anesthetized Wistar rats. We found that lymphatic segments could be divided into four phenotypes: 1) active, displaying rhythmic spontaneous contractions (51.8% of 197 analyzed sites); 2) stretch-activated, whose contraction was triggered by passive distension of the vessel lumen (4.1%); 3) passive, which displayed a completely passive distension (4.5%); and 4) inert, whose diameter never changed over time (39.6%). Smooth muscle actin was detected by immunofluorescence and confocal microscopy in the vessel walls of active but also of inert sites, albeit with a very different structure within the vessel wall. Indeed, while in active segments, actin was arranged in a dense mesh completely surrounding the lumen, in inert segments actin decorated the vessels wall in sparse longitudinal strips. When located nearby along the same lymphatic loop, active, stretch-activated, and passive sites were always recruited in temporal sequence starting from the active contraction. The time delay was ∼0.35 s between active and stretch-activated and 0.54 s between stretch-activated and passive segments, promoting a uniform lymph flux of ∼150/200 pl/min. We conclude that, unlike more central diaphragmatic lymphatic vessels, loops located at the extreme diaphragmatic periphery do require an intrinsic pumping mechanism to propel lymph centripetally, and that such an active lymph propulsion is attained by means of a complex interplay among sites whose properties differ but are indeed able to organize lymph flux in an ordered fashion. </jats:p

Compiler fuzzing: how much does it matter?

Despite much recent interest in randomised testing (fuzzing) of compilers, the practical impact of fuzzer-found compiler bugs on real-world applications has barely been assessed. We present the first quantitative and qualitative study of the tangible impact of miscompilation bugs in a mature compiler. We follow a rigorous methodology where the bug impact over the compiled application is evaluated based on (1) whether the bug appears to trigger during compilation; (2) the extent to which generated assembly code changes syntactically due to triggering of the bug; and (3) whether such changes cause regression test suite failures, or whether we can manually find application inputs that trigger execution divergence due to such changes. The study is conducted with respect to the compilation of more than 10 million lines of C/C++ code from 309 Debian packages, using 12% of the historical and now fixed miscompilation bugs found by four state-of-the-art fuzzers in the Clang/LLVM compiler, as well as 18 bugs found by human users compiling real code or as a by-product of formal verification efforts. The results show that almost half of the fuzzer-found bugs propagate to the generated binaries for at least one package, in which case only a very small part of the binary is typically affected, yet causing two failures when running the test suites of all the impacted packages. User-reported and formal verification bugs do not exhibit a higher impact, with a lower rate of triggered bugs and one test failure. The manual analysis of a selection of the syntactic changes caused by some of our bugs (fuzzer-found and non fuzzer-found) in package assembly code, shows that either these changes have no semantic impact or that they would require very specific runtime circumstances to trigger execution divergence

Interplay between gut lymphatic vessels and microbiota

Lymphatic vessels play a distinctive role in draining fluid, molecules and even cells from interstitial and serosal spaces back to the blood circulation. Lymph vessels of the gut, and especially those located in the villi (called lacteals), not only serve this primary function, but are also responsible for the transport of lipid moieties absorbed by the intestinal mucosa and serve as a second line of defence against possible bacterial infections. Here, we briefly review the current knowledge of the general mechanisms allowing lymph drainage and propulsion and will focus on the most recent findings on the mutual relationship between lacteals and intestinal microbiota

Cyclin B1-Cdk1 facilitates MAD1 release from the nuclear pore to ensure a robust spindle checkpoint.

How the cell rapidly and completely reorganizes its architecture when it divides is a problem that has fascinated researchers for almost 150 yr. We now know that the core regulatory machinery is highly conserved in eukaryotes, but how these multiple protein kinases, protein phosphatases, and ubiquitin ligases are coordinated in space and time to remodel the cell in a matter of minutes remains a major question. Cyclin B1-Cdk is the primary kinase that drives mitotic remodeling; here we show that it is targeted to the nuclear pore complex (NPC) by binding an acidic face of the kinetochore checkpoint protein, MAD1, where it coordinates NPC disassembly with kinetochore assembly. Localized cyclin B1-Cdk1 is needed for the proper release of MAD1 from the embrace of TPR at the nuclear pore so that it can be recruited to kinetochores before nuclear envelope breakdown to maintain genomic stability

Analysis of Secreted Proteins from Prepubertal Ovarian Tissues Exposed In Vitro to Cisplatin and LH

It is well known that secreted and exosomal proteins are associated with a broad range of physiological processes involving tissue homeostasis and differentiation. In the present paper, our purpose was to characterize the proteome of the culture medium in which the oocytes within the primordial/primary follicles underwent apoptosis induced by cisplatin (CIS) or were, for the most part, protected by LH against the drug. To this aim, prepubertal ovarian tissues were cultured under control and in the presence of CIS, LH, and CIS + LH. The culture media were harvested after 2, 12, and 24 h from chemotherapeutic drug treatment and analyzed by liquid chromatography–mass spectrometry (LC-MS). We found that apoptotic conditions generated by CIS in the cultured ovarian tissues and/or oocytes are reflected in distinct changes in the extracellular microenvironment in which they were cultured. These changes became evident mainly from 12 h onwards and were characterized by the inhibition or decreased release of a variety of compounds, such as the proteases Htra1 and Prss23, the antioxidants Prdx2 and Hbat1, the metabolic regulators Ldha and Pkm, and regulators of apoptotic pathways such as Tmsb4x. Altogether, these results confirm the biological relevance of the LH action on prepuberal ovaries and provide novel information about the proteins released by the ovarian tissues exposed to CIS and LH in the surrounding microenvironment. These data might represent a valuable resource for future studies aimed to clarify the effects and identify biomarkers of these compounds’ action on the developing ovary

VST: the telescope progress toward stars

The VST telescope is in an advanced stage of integration in Chile, after a period of work spent mainly on the active optics system, started in mid-2007. We present the results of the recent work on the primary and secondary mirror support systems and on the mirror cell auxiliary units

Calidad inicial y daños ocurridos durante la cosecha, transporte y lavado en batatas cultivadas en el partido de San Pedro

La batata es considerada un cultivo rústico, comparada con otras hortalizas. Sin embargo, cuando hablamos de calidad, hay muchos aspectos de manejo que deben mejorar para lograr un producto sin defectos. Un estudio previo determino que tan solo el 3,11 % de las batatas procedentes de San Pedro, no poseen ningún tipo de defecto. Como parte de las actividades realizadas en la Plataforma Herramientas de Gestión de la Calidad y del proyecto Local “Batata de calidad sampedrina”, se monitoreó la calidad de las batatas producidas en el territorio. Se plantearon dos objetivos: a) determinar la calidad inicial de la batata producida en San Pedro relevando: daños por insectos, pudriciones y defectos presentes en precosecha y b) relevar los daños ocurridos durante la cosecha, y poscosecha (transporte y lavado). Las muestras de raíces, se recolectaron en cuatro momentos: Antes de cosecha, luego de cosecha, a la llegada al galpón de acondicionado y después del lavado. En la evaluación de precosecha, o calidad inicial, se analizaron las 400 batatas (100 de cada momento de muestreo) se relevaron: daños por insectos, deformaciones, heridas cicatrizadas, brotes, venas, raicillas, batatines y otros defectos, generalmente asociados a factores genéticos y otros como heridas cicatrizadas, debido a condiciones ambientales y de manejo. Los daños poscosecha (roturas, heridas abiertas, peladuras y abrasiones se relevaron sobre 100 batatas correspondientes a cada momento de muestreo. Los defectos y daños en las raíces se relevaron según lo especificado en la Resolución SAyG Nº 297/1983 y en bibliografía existente. Los resultados mostraron que todas las raíces presentaron daños por insectos, el 46 % de las batatas presentó algún grado de deformación, la presencia de venas y batatines, fue menor. Y en cuanto a heridas abiertas, en cosecha se observaron un mínimo de 18% y un máximo de 32 % de heridas abiertas, y el 70 % de las batatas analizadas luego del lavado, presentaron heridas abiertas. Se observó que no aumentan las roturas luego de la evaluación pos cosecha, por lo que aparentemente, es durante la cosecha en el que se producen la mayor parte de ellas. A modo de conclusión se pueden agrupar los problemas de calidad según tres orígenes. De origen genético: constricciones, deformaciones, surcos, venas y batatines, estaban presentes en el 63,75 % de la muestra. Así, partiendo de material seleccionado podemos disminuir estos defectos. Daños originados por insectos: El 100 % de las batatas analizadas presento daños por insectos. Combinando prácticas de manejo, se podría reducir el ataque de las principales plagas del cultivo. Los daños originados en poscosecha: las heridas sin cicatrizar acumuladas luego del lavado, representaron el 70% de las raíces, mientras que las abrasiones fueron en su mayoría en el momento de cosecha, posiblemente asociado a la “madurez” de las raíces y a las condiciones de poca humedad del suelo

Comprehensive longitudinal non-invasive quantification of healthspan and frailty in a large cohort (n = 546) of geriatric C57BL/6 J mice

Frailty is an age-related condition characterized by a multisystem functional decline, increased vulnerability to stressors, and adverse health outcomes. Quantifying the degree of frailty in humans and animals is a health measure useful for translational geroscience research. Two frailty measurements, namely the frailty phenotype (FP) and the clinical frailty index (CFI), have been validated in mice and are frequently applied in preclinical research. However, these two tools are based on different concepts and do not necessarily identify the same mice as frail. In particular, the FP is based on a dichotomous classification that suffers from high sample size requirements and misclassification problems. Based on the monthly longitudinal non-invasive assessment of frailty in a large cohort of mice, here we develop an alternative scoring method, which we called physical function score (PFS), proposed as a continuous variable that resumes into a unique function, the five criteria included in the FP. This score would not only reduce misclassification of frailty but it also makes the two tools, PFS and CFI, integrable to provide an overall measurement of health, named vitality score (VS) in aging mice. VS displays a higher association with mortality than PFS or CFI and correlates with biomarkers related to the accumulation of senescent cells and the epigenetic clock. This longitudinal non-invasive assessment strategy and the VS may help to overcome the different sensitivity in frailty identification, reduce the sample size in longitudinal experiments, and establish the effectiveness of therapeutic/preventive interventions for frailty or other age-related diseases in geriatric animals

Desmoglein 2 is a substrate of kallikrein 7 in pancreatic cancer

<p>Abstract</p> <p>Background</p> <p>In a previous report we have demonstrated that the chymotryptic-like serine protease kallikrein 7 (<it>KLK7</it>/hK7) is overexpressed in pancreatic cancer. In normal skin, hK7 is thought to participate in skin desquamation by contributing in the degradation of desmosomal components, such as desmogleins. Thus, the ability of hK7 to degrade desmogleins was assessed and the effect of hK7 expression on desmoglein 2 was examined in cultured pancreatic cancer cells.</p> <p>Methods</p> <p>The expression of Dsg1, Dsg2, and Dsg3 in pancreatic tissues was examined by immunohistochemistry and their expression in two pancreatic cancer cell lines, BxPC-3 and Panc-1, was determined by western blot analysis. The ability of hK7 to degrade Dsg1 and Dsg2 was investigated using <it>in vitro </it>degradation assays. BxPC-3 cells stably transfected to overexpress hK7 were used to examine the effect of hK7 on cell-surface resident Dsg2.</p> <p>Results</p> <p>The levels of immunoreactive Dsg1 and Dsg2 were reduced in pancreatic adenocarcinomas compared with both normal pancreatic and chronic pancreatitis tissues. Among the desmosomal proteins examined, Dsg2 exhibited robust expression on the surface of BxPC-3 cells. When hK7 was overexpressed in this cell line, there was a significant increase in the amount of soluble Dsg2 released into the culture medium compared with vector-transfected control cells.</p> <p>Conclusion</p> <p>A reduction in the amount of the cell adhesion components Dsg1 and Dsg2 in pancreatic tumors suggests that loss of these desmosomal proteins may play a role in pancreatic cancer invasion. Using <it>in vitro </it>degradation assays, both Dsg1 and Dsg2 could be readily proteolyzed by hK7, which is overexpressed in pancreatic adenocarcinomas. The enforced expression of hK7 in BxPC-3 cells that express significant amounts of Dsg2 resulted in a marked increase in the shedding of soluble Dsg2, which is consistent with the notion that aberrant expression of hK7 in pancreatic tumors may result in diminished cell-cell adhesion and facilitate tumor cell invasion.</p